AS Pseudo Claim Charting Due Diligence US8058069B2

Purpose: Invalidity Analysis

Patent: US8058069B2
Filed: 2008-04-15
Issued: 2011-11-15
Patent Holder: (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp
Inventor(s): Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan

Title: Lipid formulations for nucleic acid delivery

Abstract: The present invention provides novel, stable lipid particles comprising one or more active agents or therapeutic agents, methods of making the lipid particles, and methods of delivering and/or administering the lipid particles. More particularly, the present invention provides stable nucleic acid-lipid particles (SNALP) comprising a nucleic acid (such as one or more interfering RNA), methods of making the SNALP, and methods of delivering and/or administering the SNALP.

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Ground	References	Owner of the Reference	Title	Semantic Mapping	Challenged Claims
Ground	References	Owner of the Reference	Title	Semantic Mapping	1	2	8	9	10	13	14	15	16	17	18	19	20	21	22
1	DNA AND CELL BIOLOGY. 12 (6): 553-560 JUL-AUG 1993 (Barthel, 1993)	Faculté de Pharmacie - Université de Strasbourg, Institut de Physiologie, URA 1446-CNRS	GENE-TRANSFER OPTIMIZATION WITH LIPOSPERMINE-COATED DNA	▪ cationic lipid ≈ cationic lipid ▪ nucleic acid ≈ nucleic acid	X	X	X				X		X	X	X
2	ANGEWANDTE CHEMIE-INTERNATIONAL EDITION IN ENGLISH. 30 (6): 613-629 JUN 1991 (Englisch, 1991)	Max Planck Institute - Experimental Medicine	CHEMICALLY MODIFIED OLIGONUCLEOTIDES AS PROBES AND INHIBITORS	▪ nucleic acid, nucleic acid mass ratio ≈ stranded nucleic acid ▪ lipid conjugate ≈ modified nucleotides	X	X							X	X	X
3	BIOTECHNIQUES. 10 (4): 520-525 APR 1991 (Rose, 1991)	Yale University	A NEW CATIONIC LIPOSOME REAGENT MEDIATING NEARLY QUANTITATIVE TRANSFECTION OF ANIMAL-CELLS	▪ nucleic acid ≈ expression system ▪ cationic lipid ≈ cationic lipid ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ neutral lipid	X	X	X	X			X		X	X	X
4	US20060051405A1 (Ian Maclachlan, 2006)	(Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Protiva Biotherapeutics Inc	Compositions for the delivery of therapeutic agents and uses thereof	▪ nucleic acid ≈ said nucleic acid ▪ lipid particle ≈ lipid particle ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ neutral lipid	X	X		X					X	X	X
5	US20060008910A1 (Ian Maclachlan, 2006)	(Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) PROTIVIA BIOTHERAPEUTICS Inc ; Arbutus Biopharma Corp	Lipid encapsulated interfering RNA	▪ inhibits aggregation ≈ inhibits aggregation ▪ nucleic acid ≈ said nucleic acid ▪ median diameter, lipid particle ≈ median diameter ▪ lipid conjugate ≈ lipid conjugate ▪ cationic lipid ≈ cationic lipid	X	X	X		X		X	X	X	X	X	X
6	WO2005120152A2 (Ian Maclachlan, 2005)	(Original Assignee) Protiva Biotherapeutics, Inc.	Cationic lipids and methods of use	▪ total lipid present ≈ total lipid present ▪ nucleic acid ≈ said nucleic acid ▪ lipid conjugate ≈ lipid conjugate ▪ DAA conjugate ≈ DAA conjugate	X	X	X					X	X	X	X		X	X
7	CN1968714A (O·凯尔, 2007)	(Original Assignee) 阿图根股份公司	脂质、脂质复合物及其应用	▪ cationic lipid ≈ 结肠癌和 ▪ nucleic acid mass ratio ≈ 摩尔比率 ▪ small interfering ≈ 反应物 ▪ pharmaceutical composition ≈ 物活性	X	X	X				X				X				X
8	WO2005105152A2 (Oliver Keil, 2005)	(Original Assignee) Atugen Ag	Lipids, lipid complexes and use thereof	▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ lipid component ▪ cationic lipid ≈ cationic lipid ▪ nucleic acid mass ratio ≈ molar ratio	X		X	X			X				X
9	US20050282188A1 (Peter Haeberli, 2005)	(Original Assignee) Sirna Therapeutics Inc (Current Assignee) Sirna Therapeutics Inc	RNA interference mediated inhibition of gene expression using short interfering nucleic acid (siNA)	▪ derivative thereof ≈ inflammatory cytokines ▪ lipid conjugate ≈ modified nucleotides ▪ nucleic acid ≈ nucleic acid	X	X					X		X	X	X			X
10	US20050118253A1 (Ian Maclachlan, 2005)	(Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Protiva Biotherapeutics Inc	Systemic delivery of serum stable plasmid lipid particles for cancer therapy	▪ lipid conjugate ≈ lipid conjugate, said prodrug ▪ cationic lipid ≈ cationic lipid ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ neutral lipid ▪ lipid particle ≈ uniform size	X		X	X			X
11	WO2005026372A1 (James Heyes, 2005)	(Original Assignee) Protiva Biotherapeutics, Inc.	Polyethyleneglycol-modified lipid compounds and uses thereof	▪ average molecular weight ≈ average molecular weight ▪ total lipid present ≈ total lipid present ▪ nucleic acid ≈ said nucleic acid ▪ DAA conjugate ≈ DAA conjugate ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ neutral lipid	X	X	X	X		X		X	X	X	X		X	X
12	US20050064595A1 (Ian Maclachlan, 2005)	(Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Protiva Biotherapeutics Inc	Lipid encapsulated interfering RNA	▪ nucleic acid ≈ said nucleic acid ▪ median diameter, lipid particle ≈ median diameter ▪ lipid conjugate ≈ lipid conjugate	X	X							X	X	X	X
13	US20040142892A1 (John Finn, 2004)	(Original Assignee) University of British Columbia (Current Assignee) University of British Columbia	Autogene nucleic acids encoding a secretable RNA polymerase	▪ total lipid present ≈ total lipid present ▪ nucleic acid ≈ said nucleic acid ▪ lipid conjugate ≈ lipid conjugate ▪ cationic lipid ≈ cationic lipid ▪ lipid particle ≈ lipid particle	X	X	X				X	X	X	X	X		X	X
14	US20040063654A1 (Mark Davis, 2004)	(Original Assignee) Insert Therapeutics Inc (Current Assignee) Insert Therapeutics Inc	Methods and compositions for therapeutic use of RNA interference	▪ lipid particle ≈ biodegradable poly ▪ nucleic acid ≈ nucleic acid	X	X							X	X	X
15	US6815432B2 (Jeffery J. Wheeler, 2004)	(Original Assignee) Inex Pharmaceuticals Corp (Current Assignee) University of British Columbia ; Arbutus Biopharma Corp	Methods for encapsulating plasmids in lipid bilayers	▪ total lipid present ≈ total lipid present ▪ nucleic acid ≈ said nucleic acid ▪ lipid conjugate ≈ lipid conjugate	X	X	X					X	X	X	X		X	X
16	US20030216335A1 (Jennifer Lockridge, 2003)	(Original Assignee) Jennifer Lockridge; Pamela Pavco; Gilad Gordon	Method and reagent for the modulation of female reproductive diseases and conditions	▪ nucleic acid ≈ said nucleic acid ▪ cationic lipid ≈ cationic lipid	X	X	X				X		X	X	X
17	US20030082103A1 (Charles Wartchow, 2003)	(Original Assignee) Targesome Inc (Current Assignee) Nanovalent Pharmaceuticals Inc	Targeted therapeutic lipid constructs having cell surface targets	▪ derivative thereof ≈ EGF receptors ▪ nucleic acid ≈ nucleic acid	X	X					X		X	X	X			X
18	US20030026831A1 (Aparna Lakkaraju, 2003)	(Original Assignee) University of Minnesota (Current Assignee) University of Minnesota	Anionic liposomes for delivery of bioactive agents	▪ lipid particle ≈ mean diameter ▪ nucleic acid ≈ nucleic acid ▪ nucleic acid mass ratio ≈ molar ratio	X	X							X	X	X
19	US20030069173A1 (Pamela Hawley-Nelson, 2003)	(Original Assignee) Life Technologies Inc (Current Assignee) Life Technologies Corp	Peptide-enhanced transfections	▪ nucleic acid ≈ said nucleic acid ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ neutral lipid	X	X		X					X	X	X
20	US20030072794A1 (Teni Boulikas, 2003)	(Original Assignee) Teni Boulikas (Current Assignee) Regulon Inc	Encapsulation of plasmid DNA (lipogenes™) and therapeutic agents with nuclear localization signal/fusogenic peptide conjugates into targeted liposome complexes	▪ derivative thereof ≈ derivative thereof ▪ lipid conjugate ≈ peptide conjugates ▪ cationic lipid ≈ cationic lipid ▪ total lipid present ≈ mole percent ▪ nucleic acid mass ratio ≈ molar ratio	X		X				X	X			X		X	X
21	US6858224B2 (Jeffrey Wheeler, 2005)	(Original Assignee) Inex Pharmaceuticals Corp (Current Assignee) University of British Columbia ; Arbutus Biopharma Corp	Method of preventing aggregation of a lipid:nucleic acid complex	▪ nucleic acid ≈ said nucleic acid ▪ cationic lipid ≈ cationic lipid	X	X	X				X		X	X	X
22	US6696424B1 (Carl Wheeler, 2004)	(Original Assignee) Vical Inc (Current Assignee) Vical Inc	Cytofectin dimers and methods of use thereof	▪ derivative thereof ≈ derivative thereof ▪ cationic lipid ≈ cationic lipid	X		X				X							X
23	US6586410B1 (Jeffery J. Wheeler, 2003)	(Original Assignee) Inex Pharmaceuticals Corp (Current Assignee) University of British Columbia ; Arbutus Biopharma Corp	Lipid-nucleic acid particles prepared via a hydrophobic lipid-nucleic acid complex intermediate and use for gene transfer	▪ inhibits aggregation ≈ inhibits aggregation ▪ total lipid present ≈ total lipid present ▪ nucleic acid ≈ said nucleic acid ▪ median diameter, lipid particle ≈ median diameter, lipid particle ▪ cationic lipid ≈ cationic lipid	X	X	X		X		X	X	X	X	X	X	X	X
24	US6852334B1 (Pieter R. Cullis, 2005)	(Original Assignee) University of British Columbia (Current Assignee) University of British Columbia	Cationic peg-lipids and methods of use	▪ derivative thereof ≈ derivative thereof ▪ lipid conjugate ≈ lipid conjugate ▪ cationic lipid ≈ cationic lipid ▪ nucleic acid ≈ nucleic acid ▪ total lipid present ≈ mole percent	X	X	X				X	X	X	X	X		X	X
25	US7166745B1 (Yongliang Chu, 2007)	(Original Assignee) Invitrogen Corp (Current Assignee) Life Technologies Corp	Transfection reagents	▪ cationic lipid ≈ cationic lipid ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ neutral lipid ▪ nucleic acid ≈ nucleic acid	X	X	X	X			X		X	X	X
26	US6534484B1 (Jeffery J. Wheeler, 2003)	(Original Assignee) Inex Pharmaceuticals Corp (Current Assignee) University of British Columbia ; Arbutus Biopharma Corp	Methods for encapsulating plasmids in lipid bilayers	▪ nucleic acid ≈ said nucleic acid ▪ cationic lipid ≈ cationic lipid ▪ lipid particle ≈ lipid particle	X	X	X				X		X	X	X
27	US6406705B1 (Heather L. Davis, 2002)	(Original Assignee) Ottawa Hospital Research Institute; Coley Pharmaceutical GmbH; University of Iowa Research Foundation UIRF (Current Assignee) Ottawa Hospital Research Institute ; Coley Pharmaceutical GmbH ; University of Iowa Research Foundation UIRF	Use of nucleic acids containing unmethylated CpG dinucleotide as an adjuvant	▪ total lipid ≈ unmethylated CpG ▪ nucleic acid ≈ nucleic acid	X	X	X					X	X	X	X		X	X
28	CA2271582A1 (Sean C. Semple, 1999)	(Original Assignee) Sean C. Semple; Sandra K. Klimuk; Troy Harasym; Michael J. Hope; Steven M. Ansell; Pieter R. Cullis; Peter Scherrer; Wilson W. K. Mok; Inex Pharmaceuticals Corp.; University Of British Columbia (Current Assignee) University of British Columbia	Method for administration of therapeutic agents, including antisense, with repeat dosing	▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ lipid component, neutral lipid ▪ lipid particle ≈ lipid particle				X
29	US6110745A (Yuan-Peng Zhang, 2000)	(Original Assignee) Inex Pharmaceuticals Corp (Current Assignee) Arbutus Biopharma Corp	Preparation of lipid-nucleic acid particles using a solvent extraction and direct hydration method	▪ nucleic acid ≈ said nucleic acid ▪ lipid conjugate ≈ lipid conjugate ▪ cationic lipid ≈ cationic lipid ▪ lipid particle ≈ lipid particle ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ neutral lipid ▪ total lipid present ≈ mole percent	X	X	X	X			X	X	X	X	X		X	X
30	US6284267B1 (Rajindra Aneja, 2001)	(Original Assignee) Nutrimed Biotech (Current Assignee) Nutrimed Biotech	Amphiphilic materials and liposome formulations thereof	▪ average molecular weight ≈ average molecular weight ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ lipid component ▪ cationic lipid ≈ cationic lipid ▪ nucleic acid ≈ nucleic acid	X	X	X	X		X	X		X	X	X
31	US5976567A (Jeffery J. Wheeler, 1999)	(Original Assignee) Inex Pharmaceuticals Corp (Current Assignee) University of British Columbia ; Arbutus Biopharma Corp	Lipid-nucleic acid particles prepared via a hydrophobic lipid-nucleic acid complex intermediate and use for gene transfer	▪ nucleic acid ≈ said nucleic acid ▪ lipid conjugate ≈ lipid conjugate ▪ lipid particle ≈ lipid particle	X	X							X	X	X
32	US5705385A (Marcel B. Bally, 1998)	(Original Assignee) Inex Pharmaceuticals Corp (Current Assignee) University of British Columbia ; Arbutus Biopharma Corp	Lipid-nucleic acid particles prepared via a hydrophobic lipid-nucleic acid complex intermediate and use for gene transfer	▪ nucleic acid ≈ said nucleic acid ▪ cationic lipid ≈ cationic lipid	X	X	X				X		X	X	X
33	US5756353A (Robert J. Debs, 1998)	(Original Assignee) University of California (Current Assignee) University of California	Expression of cloned genes in the lung by aerosol-and liposome-based delivery	▪ total lipid ≈ particle diameter ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ lipid component ▪ nucleic acid ≈ nucleic acid	X	X	X	X				X	X	X	X		X	X
34	US5820873A (Lewis S. L. Choi, 1998)	(Original Assignee) University of British Columbia (Current Assignee) OF BRITISH COLUMBIA THE, University of ; University of British Columbia	Polyethylene glycol modified ceramide lipids and liposome uses thereof	▪ average molecular weight ≈ average molecular weight ▪ total lipid present ≈ mole percent	X		X			X		X					X	X
35	US5885613A (John W. Holland, 1999)	(Original Assignee) University of British Columbia (Current Assignee) University of British Columbia	Bilayer stabilizing components and their use in forming programmable fusogenic liposomes	▪ cationic lipid ≈ cationic lipid ▪ lipid particle ≈ mean diameter ▪ total lipid present ≈ mole percent	X		X				X	X					X	X
36	US5627159A (PoJen Shih, 1997)	(Original Assignee) Life Technologies Inc (Current Assignee) Life Technologies Corp	Enhancement of lipid cationic transfections in the presence of serum	▪ nucleic acid ≈ said nucleic acid ▪ cationic lipid ≈ cationic lipid	X	X	X				X		X	X	X
37	US5578475A (Joel A. Jessee, 1996)	(Original Assignee) Life Technologies Inc (Current Assignee) Life Technologies Corp	Composition and methods for transfecting eukaryotic cells	▪ nucleic acid ≈ said nucleic acid ▪ average molecular weight ≈ said component	X	X				X			X	X	X
38	US5703055A (Philip L. Felgner, 1997)	(Original Assignee) Vical Inc; Wisconsin Alumni Research Foundation (Current Assignee) Vical Inc ; Wisconsin Alumni Research Foundation	Generation of antibodies through lipid mediated DNA delivery	▪ cationic lipid ≈ cationic lipid ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ neutral lipid	X		X	X			X
39	US5674908A (Alberto Haces, 1997)	(Original Assignee) Life Technologies Inc (Current Assignee) Life Technologies Corp	Highly packed polycationic ammonium, sulfonium and phosphonium lipids	▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ neutral lipid ▪ nucleic acid ≈ nucleic acid	X	X		X					X	X	X
40	US5320906A (Crispin G. S. Eley, 1994)	(Original Assignee) Vestar Inc (Current Assignee) NeXstar Pharmaceuticals Inc	Delivery vehicles with amphiphile-associated active ingredient	▪ lipid particle ≈ lipid particle ▪ nucleic acid ≈ fatty acids	X	X							X	X	X
41	US5283185A (Richard M. Epand, 1994)	(Original Assignee) McMaster University; University of Tennessee Research Foundation (Current Assignee) McMaster University ; University of Tennessee Research Foundation	Method for delivering nucleic acids into cells	▪ cationic lipid ≈ cationic lipid ▪ nucleic acid ≈ nucleic acid	X	X	X				X		X	X	X
42	US5225212A (Francis J. Martin, 1993)	(Original Assignee) Liposome Technology Inc (Current Assignee) Liposome Technology Inc	Microreservoir liposome composition and method	▪ total lipid ≈ particle diameter ▪ total lipid present ≈ mole percent	X		X					X					X	X
43	US5279833A (John K. Rose, 1994)	(Original Assignee) Yale University (Current Assignee) YALE UNIVERSITY A NON-PROFIT CT ; Yale University	Liposomal transfection of nucleic acids into animal cells	▪ cationic lipid ≈ cationic lipid ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ neutral lipid ▪ nucleic acid ≈ nucleic acid	X	X	X	X			X		X	X	X
44	US4987071A (Thomas R. Cech, 1991)	(Original Assignee) University Patents Inc (Current Assignee) University Patents Inc	RNA ribozyme polymerases, dephosphorylases, restriction endoribonucleases and methods	▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ polycytidylic acid ▪ nucleic acid ≈ nucleic acid	X	X		X					X	X	X
45	WO2007051303A1 (Ian Maclachlan, 2007)	(Original Assignee) Protiva Biotherapeutics, Inc.	MODIFIED siRNA MOLECULES AND USES THEREOF	▪ inhibits aggregation ≈ inhibits aggregation ▪ total lipid present ≈ total lipid present ▪ median diameter, lipid particle ≈ median diameter ▪ lipid conjugate ≈ lipid conjugate ▪ DAA conjugate ≈ DAA conjugate ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ neutral lipid	X		X	X	X			X				X	X	X
46	WO2007024323A2 (Joseph M. Desimone, 2007)	(Original Assignee) The University Of North Carolina At Chapel Hill; Liquidia Technologies Inc.; North Carolina State University	Nanoparticle fabrication methods, systems, and materials	▪ average molecular weight ≈ polymerization catalyst, cellulose ethers ▪ lipid particle, total lipid ≈ biodegradable poly, metal precursor ▪ nucleic acid ≈ carboxylic acids	X	X	X			X		X	X	X	X		X	X
47	EP1766035A1 (Ian Maclachlan, 2007)	(Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Protiva Biotherapeutics Inc	Lipid encapsulated interfering rna	▪ inhibits aggregation ≈ inhibits aggregation ▪ nucleic acid ≈ said nucleic acid ▪ median diameter, lipid particle ≈ median diameter ▪ lipid conjugate ≈ lipid conjugate ▪ cationic lipid ≈ cationic lipid	X	X	X		X		X	X	X	X	X	X
48	EP1781593A2 (Ian Maclachlan, 2007)	(Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Protiva Biotherapeutics Inc	Cationic lipids and methods of use	▪ total lipid present ≈ total lipid present ▪ nucleic acid ≈ said nucleic acid ▪ lipid conjugate ≈ lipid conjugate ▪ DAA conjugate ≈ DAA conjugate	X	X	X					X	X	X	X		X	X
49	WO2005121348A1 (Ian Maclachlan, 2005)	(Original Assignee) Protiva Biotherapeutics, Inc.	Lipid encapsulated interfering rna	▪ inhibits aggregation ≈ inhibits aggregation ▪ nucleic acid ≈ said nucleic acid ▪ median diameter, lipid particle ≈ median diameter ▪ lipid conjugate ≈ lipid conjugate ▪ cationic lipid ≈ cationic lipid	X	X	X		X		X	X	X	X	X	X
50	CN1882693A (J·海斯, 2006)	(Original Assignee) 普洛体维生物治疗公司	聚乙二醇修饰的脂质化合物及其应用	▪ cationic lipid, lipid conjugate ≈ 一种脂质 ▪ average molecular weight ≈ 平均分子 ▪ pharmaceutical composition ≈ 物活性 ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ 二棕榈	X		X	X		X	X								X
51	EP1664316A1 (James Heyes, 2006)	(Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Protiva Biotherapeutics Inc	Polyethyleneglycol-modified lipid compounds and uses thereof	▪ average molecular weight ≈ average molecular weight ▪ total lipid present ≈ total lipid present ▪ nucleic acid ≈ said nucleic acid ▪ DAA conjugate ≈ DAA conjugate ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ neutral lipid	X	X	X	X		X		X	X	X	X		X	X
52	WO2005005622A2 (Stephen Ray, 2005)	(Original Assignee) Ribostem Limited	Method of altering cell properties by administering rna	▪ lipid conjugate ≈ spinal cord injury ▪ nucleic acid ≈ immune function	X	X							X	X	X
53	EP1519714A1 (Cory Giesbrecht, 2005)	(Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Protiva Biotherapeutics Inc	Method and apparatus for producing liposomes	▪ nucleic acid mass ratio ≈ salt concentration ▪ nucleic acid ≈ said nucleic acid	X	X							X	X	X
54	CN1655764A (W·V·罗德里格扎, 2005)	(Original Assignee) 埃斯佩里安Luv发展公司	用于给予一定大小的脂质体治疗或预防疾病的组合物和方法	▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ 二棕榈酰磷脂, 酰丝氨酸 ▪ median diameter ≈ 平均直径 ▪ nucleic acid ≈ 与一种	X	X		X					X	X	X	X
55	WO9914346A2 (Tod M. Woolf, 1999)	(Original Assignee) Sequitur, Inc.	SENSE mRNA THERAPY	▪ cationic lipid ≈ cationic lipid ▪ nucleic acid ≈ nucleic acid	X	X	X				X		X	X	X
56	WO9914226A2 (Jesper Wengel, 1999)	(Original Assignee) Exiqon A/S	Bi- and tri-cyclic nucleoside, nucleotide and oligonucleotide analogues	▪ nucleic acid, nucleic acid mass ratio ≈ nucleic acid amplification reaction, stranded nucleic acid ▪ average molecular weight ≈ following criteria	X	X				X			X	X	X
57	US6376248B1 (Pamela Hawley-Nelson, 2002)	(Original Assignee) Life Technologies Inc (Current Assignee) Life Technologies Corp	Peptide-enhanced transfections	▪ nucleic acid ≈ said nucleic acid ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ neutral lipid	X	X		X					X	X	X
58	US6004573A (Ramesh C. Rathi, 1999)	(Original Assignee) MacroMed Inc (Current Assignee) JG CONSULTING AG ; KIM PHD SUNG WAN ; BTG International Ltd	Biodegradable low molecular weight triblock poly(lactide-co-glycolide) polyethylene glycol copolymers having reverse thermal gelation properties	▪ average molecular weight ≈ average molecular weight ▪ lipid particle ≈ biodegradable poly ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ polymeric drug ▪ total lipid present ≈ mole percent	X		X	X		X		X					X	X
59	US5789554A (Shui-on Leung, 1998)	(Original Assignee) Immunomedics Inc (Current Assignee) Immunomedics Inc	Immunoconjugates and humanized antibodies specific for B-cell lymphoma and leukemia cells	▪ lipid particle, total lipid present ≈ diagnostic reagent ▪ inhibits aggregation ≈ selecting cells	X		X		X			X					X	X
60	WO9640964A2 (Jeffery J. Wheeler, 1996)	(Original Assignee) Inex Pharmaceuticals Corporation	Lipid-nucleic acid particles prepared via a hydrophobic lipid-nucleic acid complex intermediate and use for gene transfer	▪ nucleic acid ≈ said nucleic acid ▪ lipid conjugate ≈ lipid conjugate	X	X							X	X	X
61	US5716785A (Russell N. Van Gelder, 1998)	(Original Assignee) Leland Stanford Junior University (Current Assignee) Leland Stanford Junior University	Processes for genetic manipulations using promoters	▪ lipid conjugate ≈ selected cell population ▪ nucleic acid, nucleic acid mass ratio ≈ stranded nucleic acid, molar excess ▪ average molecular weight ≈ human tissue	X	X				X			X	X	X
62	JPH0867666A (Masahiko Furue, 1996)	(Original Assignee) Lion Corp; ライオン株式会社	カロチノイド含有粉末製剤及びその製造方法	▪ pharmaceutical composition ≈ 型乳化物 ▪ small interfering ≈ カロチン ▪ total lipid ≈ の合計 ▪ nucleic acid mass ratio ≈ 重量比	X	X	X					X			X		X	X	X
63	WO9502698A1 (Joel A. Jessee, 1995)	(Original Assignee) Life Technologies, Inc.	Composition and methods for transfecting eukaryotic cells	▪ nucleic acid ≈ said nucleic acid ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ neutral lipid	X	X		X					X	X	X
64	JPH07242568A (Masao Kawamura, 1995)	(Original Assignee) Eisai Co Ltd; エーザイ株式会社	苦味隠蔽製剤	▪ cationic lipid ≈ 硫酸ナトリウム ▪ nucleic acid ≈ 溶解性	X	X	X				X		X	X	X
65	WO9324640A2 (Robert J. Debs, 1993)	(Original Assignee) The Regents Of The University Of California	Methods and compositions for in vivo gene therapy	▪ lipid conjugate ≈ tissue specific expression ▪ nucleic acid ≈ said nucleic acid ▪ cationic lipid ≈ cationic lipid	X	X	X				X		X	X	X
66	WO9324641A2 (Barrie J. Carter, 1993)	(Original Assignee) The United States Of America, As Represented By The Secretary, Department Of Health & Human Services	Adeno-associated virus with inverted terminal repeat sequences as promoter	▪ inhibits aggregation ≈ isolated nucleic acid ▪ phospholipid comprises dipalmitoylphosphatidylcholine ≈ repeat sequences	X			X	X			X

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	DNA AND CELL BIOLOGY. 12 (6): 553-560 JUL-AUG 1993 Publication Year: 1993 GENE-TRANSFER OPTIMIZATION WITH LIPOSPERMINE-COATED DNA Faculté de Pharmacie - Université de Strasbourg, Institut de Physiologie, URA 1446-CNRS Barthel, Remy, Loeffler, Behr
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	GENE-TRANSFER OPTIMIZATION WITH LIPOSPERMINE-COATED DNA . Designed synthetic DNA carriers represent an attractive alternative to the widely used calcium phosphate gene transfer technique . In this context , we developed a class of nucleic acid (nucleic acid) binding lipids , the lipopolyamines , which spontaneously condense DNA on a cationic lipid (cationic lipid) layer . The resulting nucleolipidic particles transfect most animal cells efficiently . However , compaction depends on many experimental factors , some of which have been varied here to give optimal transfection efficiency . When plasmid condensation by the lipospermine is performed in the absence of competing polyions or serum proteins , or when the gene of interest is diluted into carrier DNA , transfection efficiency is increased by 2-3 orders of magnitude . With these improvements , chloramphenicol acetyl transferase activity resulting from transfection of as little as 25 ng could easily be detected by a nonradioactive ELISA test .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) comprises a small interfering RNA (siRNA) .	GENE-TRANSFER OPTIMIZATION WITH LIPOSPERMINE-COATED DNA . Designed synthetic DNA carriers represent an attractive alternative to the widely used calcium phosphate gene transfer technique . In this context , we developed a class of nucleic acid (nucleic acid) binding lipids , the lipopolyamines , which spontaneously condense DNA on a cationic lipid layer . The resulting nucleolipidic particles transfect most animal cells efficiently . However , compaction depends on many experimental factors , some of which have been varied here to give optimal transfection efficiency . When plasmid condensation by the lipospermine is performed in the absence of competing polyions or serum proteins , or when the gene of interest is diluted into carrier DNA , transfection efficiency is increased by 2-3 orders of magnitude . With these improvements , chloramphenicol acetyl transferase activity resulting from transfection of as little as 25 ng could easily be detected by a nonradioactive ELISA test .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	GENE-TRANSFER OPTIMIZATION WITH LIPOSPERMINE-COATED DNA . Designed synthetic DNA carriers represent an attractive alternative to the widely used calcium phosphate gene transfer technique . In this context , we developed a class of nucleic acid binding lipids , the lipopolyamines , which spontaneously condense DNA on a cationic lipid (cationic lipid) layer . The resulting nucleolipidic particles transfect most animal cells efficiently . However , compaction depends on many experimental factors , some of which have been varied here to give optimal transfection efficiency . When plasmid condensation by the lipospermine is performed in the absence of competing polyions or serum proteins , or when the gene of interest is diluted into carrier DNA , transfection efficiency is increased by 2-3 orders of magnitude . With these improvements , chloramphenicol acetyl transferase activity resulting from transfection of as little as 25 ng could easily be detected by a nonradioactive ELISA test .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	GENE-TRANSFER OPTIMIZATION WITH LIPOSPERMINE-COATED DNA . Designed synthetic DNA carriers represent an attractive alternative to the widely used calcium phosphate gene transfer technique . In this context , we developed a class of nucleic acid binding lipids , the lipopolyamines , which spontaneously condense DNA on a cationic lipid (cationic lipid) layer . The resulting nucleolipidic particles transfect most animal cells efficiently . However , compaction depends on many experimental factors , some of which have been varied here to give optimal transfection efficiency . When plasmid condensation by the lipospermine is performed in the absence of competing polyions or serum proteins , or when the gene of interest is diluted into carrier DNA , transfection efficiency is increased by 2-3 orders of magnitude . With these improvements , chloramphenicol acetyl transferase activity resulting from transfection of as little as 25 ng could easily be detected by a nonradioactive ELISA test .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	GENE-TRANSFER OPTIMIZATION WITH LIPOSPERMINE-COATED DNA . Designed synthetic DNA carriers represent an attractive alternative to the widely used calcium phosphate gene transfer technique . In this context , we developed a class of nucleic acid (nucleic acid) binding lipids , the lipopolyamines , which spontaneously condense DNA on a cationic lipid layer . The resulting nucleolipidic particles transfect most animal cells efficiently . However , compaction depends on many experimental factors , some of which have been varied here to give optimal transfection efficiency . When plasmid condensation by the lipospermine is performed in the absence of competing polyions or serum proteins , or when the gene of interest is diluted into carrier DNA , transfection efficiency is increased by 2-3 orders of magnitude . With these improvements , chloramphenicol acetyl transferase activity resulting from transfection of as little as 25 ng could easily be detected by a nonradioactive ELISA test .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	GENE-TRANSFER OPTIMIZATION WITH LIPOSPERMINE-COATED DNA . Designed synthetic DNA carriers represent an attractive alternative to the widely used calcium phosphate gene transfer technique . In this context , we developed a class of nucleic acid (nucleic acid) binding lipids , the lipopolyamines , which spontaneously condense DNA on a cationic lipid layer . The resulting nucleolipidic particles transfect most animal cells efficiently . However , compaction depends on many experimental factors , some of which have been varied here to give optimal transfection efficiency . When plasmid condensation by the lipospermine is performed in the absence of competing polyions or serum proteins , or when the gene of interest is diluted into carrier DNA , transfection efficiency is increased by 2-3 orders of magnitude . With these improvements , chloramphenicol acetyl transferase activity resulting from transfection of as little as 25 ng could easily be detected by a nonradioactive ELISA test .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (nucleic acid) mass ratio of from about 5 to about 15 .	GENE-TRANSFER OPTIMIZATION WITH LIPOSPERMINE-COATED DNA . Designed synthetic DNA carriers represent an attractive alternative to the widely used calcium phosphate gene transfer technique . In this context , we developed a class of nucleic acid (nucleic acid) binding lipids , the lipopolyamines , which spontaneously condense DNA on a cationic lipid layer . The resulting nucleolipidic particles transfect most animal cells efficiently . However , compaction depends on many experimental factors , some of which have been varied here to give optimal transfection efficiency . When plasmid condensation by the lipospermine is performed in the absence of competing polyions or serum proteins , or when the gene of interest is diluted into carrier DNA , transfection efficiency is increased by 2-3 orders of magnitude . With these improvements , chloramphenicol acetyl transferase activity resulting from transfection of as little as 25 ng could easily be detected by a nonradioactive ELISA test .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	ANGEWANDTE CHEMIE-INTERNATIONAL EDITION IN ENGLISH. 30 (6): 613-629 JUN 1991 Publication Year: 1991 CHEMICALLY MODIFIED OLIGONUCLEOTIDES AS PROBES AND INHIBITORS Max Planck Institute - Experimental Medicine Englisch, Gauss
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (stranded nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	CHEMICALLY MODIFIED OLIGONUCLEOTIDES AS PROBES AND INHIBITORS . Oligonucleotides bind specifically to single-stranded nucleic acid (nucleic acid, nucleic acid mass ratio) s to form a double helix if there is a complementary antiparallel nucleotide sequence . In addition , certain oligonucleotides bind specifically to a variety of proteins . Therefore , biological processes involving these nucleic acids or proteins can be modulated (normally inhibited) by addition of the respective oligonucleotides . The effects of these oligonucleotides and the fields of potential application can be broadened by the introduction of chemically modified nucleotides . For instance , replacing oligonucleotide phosphate groups by methylphosphonates results in the loss of one negative charge per nucleotide and the oligonucleotide becomes more lipophilic . An oligonucleotide carrying a reactive group can modify its binding partner . An oligonucleotide covalently linked to a dye can be localized in a biological specimen . Oligonucleotides attached to an enzyme can be detected in very small amounts since the enzyme can catalyze the formation of large amounts of the substance assayed (e . g . , a fluorescent product) . An important biochemical application is the detection and localization of a messenger RNA or its gene . Medical applications include the detection of bacterial or viral sequences . There is also great interest in inhibiting the translation of messenger RNA and the transcription and replication of DNA . So-called antisense oligonucleotides are currently being used for the inhibition of protein biosynthesis and of reverse transcription of retroviruses .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (stranded nucleic acid) comprises a small interfering RNA (siRNA) .	CHEMICALLY MODIFIED OLIGONUCLEOTIDES AS PROBES AND INHIBITORS . Oligonucleotides bind specifically to single-stranded nucleic acid (nucleic acid, nucleic acid mass ratio) s to form a double helix if there is a complementary antiparallel nucleotide sequence . In addition , certain oligonucleotides bind specifically to a variety of proteins . Therefore , biological processes involving these nucleic acids or proteins can be modulated (normally inhibited) by addition of the respective oligonucleotides . The effects of these oligonucleotides and the fields of potential application can be broadened by the introduction of chemically modified nucleotides . For instance , replacing oligonucleotide phosphate groups by methylphosphonates results in the loss of one negative charge per nucleotide and the oligonucleotide becomes more lipophilic . An oligonucleotide carrying a reactive group can modify its binding partner . An oligonucleotide covalently linked to a dye can be localized in a biological specimen . Oligonucleotides attached to an enzyme can be detected in very small amounts since the enzyme can catalyze the formation of large amounts of the substance assayed (e . g . , a fluorescent product) . An important biochemical application is the detection and localization of a messenger RNA or its gene . Medical applications include the detection of bacterial or viral sequences . There is also great interest in inhibiting the translation of messenger RNA and the transcription and replication of DNA . So-called antisense oligonucleotides are currently being used for the inhibition of protein biosynthesis and of reverse transcription of retroviruses .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (stranded nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	CHEMICALLY MODIFIED OLIGONUCLEOTIDES AS PROBES AND INHIBITORS . Oligonucleotides bind specifically to single-stranded nucleic acid (nucleic acid, nucleic acid mass ratio) s to form a double helix if there is a complementary antiparallel nucleotide sequence . In addition , certain oligonucleotides bind specifically to a variety of proteins . Therefore , biological processes involving these nucleic acids or proteins can be modulated (normally inhibited) by addition of the respective oligonucleotides . The effects of these oligonucleotides and the fields of potential application can be broadened by the introduction of chemically modified nucleotides . For instance , replacing oligonucleotide phosphate groups by methylphosphonates results in the loss of one negative charge per nucleotide and the oligonucleotide becomes more lipophilic . An oligonucleotide carrying a reactive group can modify its binding partner . An oligonucleotide covalently linked to a dye can be localized in a biological specimen . Oligonucleotides attached to an enzyme can be detected in very small amounts since the enzyme can catalyze the formation of large amounts of the substance assayed (e . g . , a fluorescent product) . An important biochemical application is the detection and localization of a messenger RNA or its gene . Medical applications include the detection of bacterial or viral sequences . There is also great interest in inhibiting the translation of messenger RNA and the transcription and replication of DNA . So-called antisense oligonucleotides are currently being used for the inhibition of protein biosynthesis and of reverse transcription of retroviruses .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (stranded nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	CHEMICALLY MODIFIED OLIGONUCLEOTIDES AS PROBES AND INHIBITORS . Oligonucleotides bind specifically to single-stranded nucleic acid (nucleic acid, nucleic acid mass ratio) s to form a double helix if there is a complementary antiparallel nucleotide sequence . In addition , certain oligonucleotides bind specifically to a variety of proteins . Therefore , biological processes involving these nucleic acids or proteins can be modulated (normally inhibited) by addition of the respective oligonucleotides . The effects of these oligonucleotides and the fields of potential application can be broadened by the introduction of chemically modified nucleotides . For instance , replacing oligonucleotide phosphate groups by methylphosphonates results in the loss of one negative charge per nucleotide and the oligonucleotide becomes more lipophilic . An oligonucleotide carrying a reactive group can modify its binding partner . An oligonucleotide covalently linked to a dye can be localized in a biological specimen . Oligonucleotides attached to an enzyme can be detected in very small amounts since the enzyme can catalyze the formation of large amounts of the substance assayed (e . g . , a fluorescent product) . An important biochemical application is the detection and localization of a messenger RNA or its gene . Medical applications include the detection of bacterial or viral sequences . There is also great interest in inhibiting the translation of messenger RNA and the transcription and replication of DNA . So-called antisense oligonucleotides are currently being used for the inhibition of protein biosynthesis and of reverse transcription of retroviruses .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (stranded nucleic acid) mass ratio of from about 5 to about 15 .	CHEMICALLY MODIFIED OLIGONUCLEOTIDES AS PROBES AND INHIBITORS . Oligonucleotides bind specifically to single-stranded nucleic acid (nucleic acid, nucleic acid mass ratio) s to form a double helix if there is a complementary antiparallel nucleotide sequence . In addition , certain oligonucleotides bind specifically to a variety of proteins . Therefore , biological processes involving these nucleic acids or proteins can be modulated (normally inhibited) by addition of the respective oligonucleotides . The effects of these oligonucleotides and the fields of potential application can be broadened by the introduction of chemically modified nucleotides . For instance , replacing oligonucleotide phosphate groups by methylphosphonates results in the loss of one negative charge per nucleotide and the oligonucleotide becomes more lipophilic . An oligonucleotide carrying a reactive group can modify its binding partner . An oligonucleotide covalently linked to a dye can be localized in a biological specimen . Oligonucleotides attached to an enzyme can be detected in very small amounts since the enzyme can catalyze the formation of large amounts of the substance assayed (e . g . , a fluorescent product) . An important biochemical application is the detection and localization of a messenger RNA or its gene . Medical applications include the detection of bacterial or viral sequences . There is also great interest in inhibiting the translation of messenger RNA and the transcription and replication of DNA . So-called antisense oligonucleotides are currently being used for the inhibition of protein biosynthesis and of reverse transcription of retroviruses .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	BIOTECHNIQUES. 10 (4): 520-525 APR 1991 Publication Year: 1991 A NEW CATIONIC LIPOSOME REAGENT MEDIATING NEARLY QUANTITATIVE TRANSFECTION OF ANIMAL-CELLS Yale University Rose, Buonocore, Whitt
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (expression system) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	A NEW CATIONIC LIPOSOME REAGENT MEDIATING NEARLY QUANTITATIVE TRANSFECTION OF ANIMAL-CELLS . One of the most efficient systems for the expression of genes in the cytoplasm of animal cells utilizes a recombinant vaccinia virus encoding the bacteriophage T7 RNA polymerase . Cells infected with this virus are transfected with plasmid DNAs containing the gene to be expressed under T7 promoter control . The major limitation of this system is the efficiency with which DNA is introduced into the cell . Recently , a cationic liposome-mediated transfection reagent has yielded transfection frequencies of greater than 80% . To determine if commercially available cationic lipid (cationic lipid) s could form liposomes that would yield similar transfection efficiencies , we tested liposomes prepared with five different cationic lipids . When used at appropriate concentrations in liposomes that also contained a neutral lipid , four of the five cationic lipids were effective in the transfection of HeLa cells . However , liposomes formed with the neutral lipid and one of the cationic lipids , dimethyldioctadecyl-ammonium bromide (DDAB) , gave transfection frequencies of greater than 95% and had a broad spectrum of effectiveness on a variety of cell lines . Liposomes containing DDAB are an inexpensive , highly efficient and reproducible alternative for the transfection of animal cells and are well suited for use with the vaccinia virus/T7 expression system (nucleic acid) .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (expression system) comprises a small interfering RNA (siRNA) .	A NEW CATIONIC LIPOSOME REAGENT MEDIATING NEARLY QUANTITATIVE TRANSFECTION OF ANIMAL-CELLS . One of the most efficient systems for the expression of genes in the cytoplasm of animal cells utilizes a recombinant vaccinia virus encoding the bacteriophage T7 RNA polymerase . Cells infected with this virus are transfected with plasmid DNAs containing the gene to be expressed under T7 promoter control . The major limitation of this system is the efficiency with which DNA is introduced into the cell . Recently , a cationic liposome-mediated transfection reagent has yielded transfection frequencies of greater than 80% . To determine if commercially available cationic lipids could form liposomes that would yield similar transfection efficiencies , we tested liposomes prepared with five different cationic lipids . When used at appropriate concentrations in liposomes that also contained a neutral lipid , four of the five cationic lipids were effective in the transfection of HeLa cells . However , liposomes formed with the neutral lipid and one of the cationic lipids , dimethyldioctadecyl-ammonium bromide (DDAB) , gave transfection frequencies of greater than 95% and had a broad spectrum of effectiveness on a variety of cell lines . Liposomes containing DDAB are an inexpensive , highly efficient and reproducible alternative for the transfection of animal cells and are well suited for use with the vaccinia virus/T7 expression system (nucleic acid) .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	A NEW CATIONIC LIPOSOME REAGENT MEDIATING NEARLY QUANTITATIVE TRANSFECTION OF ANIMAL-CELLS . One of the most efficient systems for the expression of genes in the cytoplasm of animal cells utilizes a recombinant vaccinia virus encoding the bacteriophage T7 RNA polymerase . Cells infected with this virus are transfected with plasmid DNAs containing the gene to be expressed under T7 promoter control . The major limitation of this system is the efficiency with which DNA is introduced into the cell . Recently , a cationic liposome-mediated transfection reagent has yielded transfection frequencies of greater than 80% . To determine if commercially available cationic lipid (cationic lipid) s could form liposomes that would yield similar transfection efficiencies , we tested liposomes prepared with five different cationic lipids . When used at appropriate concentrations in liposomes that also contained a neutral lipid , four of the five cationic lipids were effective in the transfection of HeLa cells . However , liposomes formed with the neutral lipid and one of the cationic lipids , dimethyldioctadecyl-ammonium bromide (DDAB) , gave transfection frequencies of greater than 95% and had a broad spectrum of effectiveness on a variety of cell lines . Liposomes containing DDAB are an inexpensive , highly efficient and reproducible alternative for the transfection of animal cells and are well suited for use with the vaccinia virus/T7 expression system .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (neutral lipid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	A NEW CATIONIC LIPOSOME REAGENT MEDIATING NEARLY QUANTITATIVE TRANSFECTION OF ANIMAL-CELLS . One of the most efficient systems for the expression of genes in the cytoplasm of animal cells utilizes a recombinant vaccinia virus encoding the bacteriophage T7 RNA polymerase . Cells infected with this virus are transfected with plasmid DNAs containing the gene to be expressed under T7 promoter control . The major limitation of this system is the efficiency with which DNA is introduced into the cell . Recently , a cationic liposome-mediated transfection reagent has yielded transfection frequencies of greater than 80% . To determine if commercially available cationic lipids could form liposomes that would yield similar transfection efficiencies , we tested liposomes prepared with five different cationic lipids . When used at appropriate concentrations in liposomes that also contained a neutral lipid (phospholipid comprises dipalmitoylphosphatidylcholine) , four of the five cationic lipids were effective in the transfection of HeLa cells . However , liposomes formed with the neutral lipid and one of the cationic lipids , dimethyldioctadecyl-ammonium bromide (DDAB) , gave transfection frequencies of greater than 95% and had a broad spectrum of effectiveness on a variety of cell lines . Liposomes containing DDAB are an inexpensive , highly efficient and reproducible alternative for the transfection of animal cells and are well suited for use with the vaccinia virus/T7 expression system .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	A NEW CATIONIC LIPOSOME REAGENT MEDIATING NEARLY QUANTITATIVE TRANSFECTION OF ANIMAL-CELLS . One of the most efficient systems for the expression of genes in the cytoplasm of animal cells utilizes a recombinant vaccinia virus encoding the bacteriophage T7 RNA polymerase . Cells infected with this virus are transfected with plasmid DNAs containing the gene to be expressed under T7 promoter control . The major limitation of this system is the efficiency with which DNA is introduced into the cell . Recently , a cationic liposome-mediated transfection reagent has yielded transfection frequencies of greater than 80% . To determine if commercially available cationic lipid (cationic lipid) s could form liposomes that would yield similar transfection efficiencies , we tested liposomes prepared with five different cationic lipids . When used at appropriate concentrations in liposomes that also contained a neutral lipid , four of the five cationic lipids were effective in the transfection of HeLa cells . However , liposomes formed with the neutral lipid and one of the cationic lipids , dimethyldioctadecyl-ammonium bromide (DDAB) , gave transfection frequencies of greater than 95% and had a broad spectrum of effectiveness on a variety of cell lines . Liposomes containing DDAB are an inexpensive , highly efficient and reproducible alternative for the transfection of animal cells and are well suited for use with the vaccinia virus/T7 expression system .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (expression system) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	A NEW CATIONIC LIPOSOME REAGENT MEDIATING NEARLY QUANTITATIVE TRANSFECTION OF ANIMAL-CELLS . One of the most efficient systems for the expression of genes in the cytoplasm of animal cells utilizes a recombinant vaccinia virus encoding the bacteriophage T7 RNA polymerase . Cells infected with this virus are transfected with plasmid DNAs containing the gene to be expressed under T7 promoter control . The major limitation of this system is the efficiency with which DNA is introduced into the cell . Recently , a cationic liposome-mediated transfection reagent has yielded transfection frequencies of greater than 80% . To determine if commercially available cationic lipids could form liposomes that would yield similar transfection efficiencies , we tested liposomes prepared with five different cationic lipids . When used at appropriate concentrations in liposomes that also contained a neutral lipid , four of the five cationic lipids were effective in the transfection of HeLa cells . However , liposomes formed with the neutral lipid and one of the cationic lipids , dimethyldioctadecyl-ammonium bromide (DDAB) , gave transfection frequencies of greater than 95% and had a broad spectrum of effectiveness on a variety of cell lines . Liposomes containing DDAB are an inexpensive , highly efficient and reproducible alternative for the transfection of animal cells and are well suited for use with the vaccinia virus/T7 expression system (nucleic acid) .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (expression system) is fully encapsulated in the nucleic acid-lipid particle .	A NEW CATIONIC LIPOSOME REAGENT MEDIATING NEARLY QUANTITATIVE TRANSFECTION OF ANIMAL-CELLS . One of the most efficient systems for the expression of genes in the cytoplasm of animal cells utilizes a recombinant vaccinia virus encoding the bacteriophage T7 RNA polymerase . Cells infected with this virus are transfected with plasmid DNAs containing the gene to be expressed under T7 promoter control . The major limitation of this system is the efficiency with which DNA is introduced into the cell . Recently , a cationic liposome-mediated transfection reagent has yielded transfection frequencies of greater than 80% . To determine if commercially available cationic lipids could form liposomes that would yield similar transfection efficiencies , we tested liposomes prepared with five different cationic lipids . When used at appropriate concentrations in liposomes that also contained a neutral lipid , four of the five cationic lipids were effective in the transfection of HeLa cells . However , liposomes formed with the neutral lipid and one of the cationic lipids , dimethyldioctadecyl-ammonium bromide (DDAB) , gave transfection frequencies of greater than 95% and had a broad spectrum of effectiveness on a variety of cell lines . Liposomes containing DDAB are an inexpensive , highly efficient and reproducible alternative for the transfection of animal cells and are well suited for use with the vaccinia virus/T7 expression system (nucleic acid) .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (expression system) mass ratio of from about 5 to about 15 .	A NEW CATIONIC LIPOSOME REAGENT MEDIATING NEARLY QUANTITATIVE TRANSFECTION OF ANIMAL-CELLS . One of the most efficient systems for the expression of genes in the cytoplasm of animal cells utilizes a recombinant vaccinia virus encoding the bacteriophage T7 RNA polymerase . Cells infected with this virus are transfected with plasmid DNAs containing the gene to be expressed under T7 promoter control . The major limitation of this system is the efficiency with which DNA is introduced into the cell . Recently , a cationic liposome-mediated transfection reagent has yielded transfection frequencies of greater than 80% . To determine if commercially available cationic lipids could form liposomes that would yield similar transfection efficiencies , we tested liposomes prepared with five different cationic lipids . When used at appropriate concentrations in liposomes that also contained a neutral lipid , four of the five cationic lipids were effective in the transfection of HeLa cells . However , liposomes formed with the neutral lipid and one of the cationic lipids , dimethyldioctadecyl-ammonium bromide (DDAB) , gave transfection frequencies of greater than 95% and had a broad spectrum of effectiveness on a variety of cell lines . Liposomes containing DDAB are an inexpensive , highly efficient and reproducible alternative for the transfection of animal cells and are well suited for use with the vaccinia virus/T7 expression system (nucleic acid) .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US20060051405A1 Filed: 2005-07-19 Issued: 2006-03-09 Compositions for the delivery of therapeutic agents and uses thereof (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Protiva Biotherapeutics Inc Ian Maclachlan, Lloyd Jeffs
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US20060051405A1 CLAIM 22 . The method in accordance with claim 1 , wherein said nucleic acid (nucleic acid) -lipid particle further comprises cholesterol .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	US20060051405A1 CLAIM 22 . The method in accordance with claim 1 , wherein said nucleic acid (nucleic acid) -lipid particle further comprises cholesterol .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (neutral lipid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	US20060051405A1 CLAIM 6 . The method in accordance with claim 1 , wherein said noncationic lipid is a neutral lipid (phospholipid comprises dipalmitoylphosphatidylcholine) .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US20060051405A1 CLAIM 22 . The method in accordance with claim 1 , wherein said nucleic acid (nucleic acid) -lipid particle further comprises cholesterol .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US20060051405A1 CLAIM 22 . The method in accordance with claim 1 , wherein said nucleic acid (nucleic acid) -lipid particle further comprises cholesterol .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	US20060051405A1 CLAIM 22 . The method in accordance with claim 1 , wherein said nucleic acid (nucleic acid) -lipid particle further comprises cholesterol .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US20060008910A1 Filed: 2005-06-07 Issued: 2006-01-12 Lipid encapsulated interfering RNA (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) PROTIVIA BIOTHERAPEUTICS Inc ; Arbutus Biopharma Corp Ian Maclachlan, Lorne Palmer, James Heyes
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US20060008910A1 CLAIM 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid (cationic lipid) of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C 1 -C 3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	US20060008910A1 CLAIM 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C 1 -C 3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	US20060008910A1 CLAIM 1 . An nucleic acid-lipid particle , said nucleic acid-lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid (cationic lipid) of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C 1 -C 3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles .
US8058069B2 CLAIM 10 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation (inhibits aggregation) of particles comprises a polyethyleneglycol (PEG)-lipid conjugate .	US20060008910A1 CLAIM 1 . An nucleic acid-lipid particle , said nucleic acid-lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C 1 -C 3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	US20060008910A1 CLAIM 1 . An nucleic acid-lipid particle , said nucleic acid-lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid (cationic lipid) of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C 1 -C 3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation (inhibits aggregation) of particles comprises from 1 mol % to 2 mol % of the total lipid present in the particle .	US20060008910A1 CLAIM 1 . An nucleic acid-lipid particle , said nucleic acid-lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C 1 -C 3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US20060008910A1 CLAIM 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C 1 -C 3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US20060008910A1 CLAIM 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C 1 -C 3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	US20060008910A1 CLAIM 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C 1 -C 3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles .
US8058069B2 CLAIM 19 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a median diameter (median diameter) of from about 40 nm to about 150 nm .	US20060008910A1 CLAIM 4 . The nucleic acid-lipid particle of claim 1 , wherein said particle has a median diameter (median diameter, lipid particle) of less than about 150 nm .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	WO2005120152A2 Filed: 2005-06-07 Issued: 2005-12-22 Cationic lipids and methods of use (Original Assignee) Protiva Biotherapeutics, Inc. Ian Maclachlan, James Heyes, Lorne R. Palmer
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present (total lipid present) in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	WO2005120152A2 CLAIM 27 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said nucleic acid (nucleic acid) is a member selected from the group consisting of : DNA , RNA , siRNA , a plasmid , an antisense oligonucleotide , a ribozyme . WO2005120152A2 CLAIM 32 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	WO2005120152A2 CLAIM 27 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said nucleic acid (nucleic acid) is a member selected from the group consisting of : DNA , RNA , siRNA , a plasmid , an antisense oligonucleotide , a ribozyme .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 52 mol % to 62 mol % of the total lipid present (total lipid present) in the particle .	WO2005120152A2 CLAIM 32 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid present (total lipid present) in the particle .	WO2005120152A2 CLAIM 32 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	WO2005120152A2 CLAIM 27 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said nucleic acid (nucleic acid) is a member selected from the group consisting of : DNA , RNA , siRNA , a plasmid , an antisense oligonucleotide , a ribozyme .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	WO2005120152A2 CLAIM 27 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said nucleic acid (nucleic acid) is a member selected from the group consisting of : DNA , RNA , siRNA , a plasmid , an antisense oligonucleotide , a ribozyme .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	WO2005120152A2 CLAIM 27 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said nucleic acid (nucleic acid) is a member selected from the group consisting of : DNA , RNA , siRNA , a plasmid , an antisense oligonucleotide , a ribozyme .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present (total lipid present) in the particle .	WO2005120152A2 CLAIM 32 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid present (total lipid present) in the particle .	WO2005120152A2 CLAIM 32 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	CN1968714A Filed: 2005-05-06 Issued: 2007-05-23 脂质、脂质复合物及其应用 (Original Assignee) 阿图根股份公司 O·凯尔, J·考夫曼
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid ; (b) a cationic lipid (结肠癌和) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	CN1968714A CLAIM 37 . 按照权利要求36的应用，其中所述药物用于治疗癌症，其中优选地所述癌症选自包括实体和非实体肿瘤的组，并且其中更优选地所述实体瘤选自包括下列各项的组：胰腺癌、乳腺癌、前列腺癌、肺癌、结肠癌和 (cationic lipid) 肝细胞癌。
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid comprises a small interfering (反应物) RNA (siRNA) .	CN1968714A CLAIM 56 . 按照权利要求54和55的方法，其中所述反应物 (small interfering) 还与酸清除剂反应，其中所述酸清除剂优选地选自包括三乙胺、二异丙基乙胺和吡啶的组。
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (结肠癌和) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	CN1968714A CLAIM 37 . 按照权利要求36的应用，其中所述药物用于治疗癌症，其中优选地所述癌症选自包括实体和非实体肿瘤的组，并且其中更优选地所述实体瘤选自包括下列各项的组：胰腺癌、乳腺癌、前列腺癌、肺癌、结肠癌和 (cationic lipid) 肝细胞癌。
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (结肠癌和) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	CN1968714A CLAIM 37 . 按照权利要求36的应用，其中所述药物用于治疗癌症，其中优选地所述癌症选自包括实体和非实体肿瘤的组，并且其中更优选地所述实体瘤选自包括下列各项的组：胰腺癌、乳腺癌、前列腺癌、肺癌、结肠癌和 (cationic lipid) 肝细胞癌。
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid mass ratio (摩尔比率) of from about 5 to about 15 .	CN1968714A CLAIM 29 . 按照权利要求8-28任一项的组合物，其中所述功能性核酸是双链核糖核酸，其中所述组合物还包含核酸，优选地功能性核酸，其更优选地是双链核糖核酸，并且更优选地是选自包括下列各项的组的核酸：RNAi、siRNA、siNA、反义核酸和核酶，其中优选地RNAi与阳离子脂质的摩尔比率 (nucleic acid mass ratio) 是约0-0 . 075，优选地约0 . 02-0 . 05并且甚至更优选地0 . 037。
US8058069B2 CLAIM 22 . A pharmaceutical composition (物活性) comprising a nucleic acid-lipid particle of claim 1 and a pharmaceutically acceptable carrier .	CN1968714A CLAIM 10 . 一种药物组合物，其包括按照权利要求1-7任一项的化合物和药物活性 (pharmaceutical composition) 化合物以及优选地药用载体。

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	WO2005105152A2 Filed: 2005-05-06 Issued: 2005-11-10 Lipids, lipid complexes and use thereof (Original Assignee) Atugen Ag Oliver Keil, Jörg Kaufmann
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	WO2005105152A2 CLAIM 5 . The compound according to any of claims 1 to 4 , wherein the compound is a cationic lipid (cationic lipid) , preferably in association with an anion Y " ; .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	WO2005105152A2 CLAIM 5 . The compound according to any of claims 1 to 4 , wherein the compound is a cationic lipid (cationic lipid) , preferably in association with an anion Y " ; .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (lipid component) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	WO2005105152A2 CLAIM 8 . A composition comprising as a lipid component (phospholipid comprises dipalmitoylphosphatidylcholine) a compound according to any of claims 1 to 7 , and a carrier .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	WO2005105152A2 CLAIM 5 . The compound according to any of claims 1 to 4 , wherein the compound is a cationic lipid (cationic lipid) , preferably in association with an anion Y " ; .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid mass ratio (molar ratio) of from about 5 to about 15 .	WO2005105152A2 CLAIM 29 . The composition according to any of claim 8 to 28 , wherein the functional nucleic acid is a double-sfranded ribonucleic acid , wherein the composition further comprises a nucleic acid , preferably a functional nucleic acid which is more preferably a double-sfranded ribonucleic acid and most preferably a nucleic acid selected from the group comprising RNAi , siRNA , siNA , antisense nucleic acid and ribozyme , whereby preferably the molar ratio (nucleic acid mass ratio) n of RNAi to cationic lipid is from about 0 to 0 . 075 , preferably from about 0 . 02 to 0 . 05 and even more preferably 0 . 037 .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US20050282188A1 Filed: 2005-04-04 Issued: 2005-12-22 RNA interference mediated inhibition of gene expression using short interfering nucleic acid (siNA) (Original Assignee) Sirna Therapeutics Inc (Current Assignee) Sirna Therapeutics Inc Peter Haeberli, Leonid Beigelman
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof (inflammatory cytokines) , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US20050282188A1 CLAIM 1 . A chemically synthesized double stranded short interfering nucleic acid (nucleic acid) (siNA) molecule that directs cleavage of a target RNA via RNA interference (RNAi) , wherein : a . each strand of said siNA molecule is about 18 to about 38 nucleotides in length ; b . one strand of said siNA molecule comprises nucleotide sequence having sufficient complementarity to said target RNA for the siNA molecule to direct cleavage of the target RNA via RNA interference ; and c . wherein one or more nucleotides of said siNA molecule are chemically modified to reduce the immunostimulatory properties of the siNA molecule to a level below that of a corresponding siNA molecule having unmodified nucleotides . US20050282188A1 CLAIM 33 . The method of claim 32 , wherein said reduced immunostimulatory properties comprise an abrogated or reduced induction of inflammatory or proinflammatory cytokines (derivative thereof) in response to the siNA being introduced in a cell , tissue , or organism .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) comprises a small interfering RNA (siRNA) .	US20050282188A1 CLAIM 1 . A chemically synthesized double stranded short interfering nucleic acid (nucleic acid) (siNA) molecule that directs cleavage of a target RNA via RNA interference (RNAi) , wherein : a . each strand of said siNA molecule is about 18 to about 38 nucleotides in length ; b . one strand of said siNA molecule comprises nucleotide sequence having sufficient complementarity to said target RNA for the siNA molecule to direct cleavage of the target RNA via RNA interference ; and c . wherein one or more nucleotides of said siNA molecule are chemically modified to reduce the immunostimulatory properties of the siNA molecule to a level below that of a corresponding siNA molecule having unmodified nucleotides .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof (inflammatory cytokines) , and about 1 . 4 mol % PEG-lipid conjugate .	US20050282188A1 CLAIM 33 . The method of claim 32 , wherein said reduced immunostimulatory properties comprise an abrogated or reduced induction of inflammatory or proinflammatory cytokines (derivative thereof) in response to the siNA being introduced in a cell , tissue , or organism .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US20050282188A1 CLAIM 1 . A chemically synthesized double stranded short interfering nucleic acid (nucleic acid) (siNA) molecule that directs cleavage of a target RNA via RNA interference (RNAi) , wherein : a . each strand of said siNA molecule is about 18 to about 38 nucleotides in length ; b . one strand of said siNA molecule comprises nucleotide sequence having sufficient complementarity to said target RNA for the siNA molecule to direct cleavage of the target RNA via RNA interference ; and c . wherein one or more nucleotides of said siNA molecule are chemically modified to reduce the immunostimulatory properties of the siNA molecule to a level below that of a corresponding siNA molecule having unmodified nucleotides .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US20050282188A1 CLAIM 1 . A chemically synthesized double stranded short interfering nucleic acid (nucleic acid) (siNA) molecule that directs cleavage of a target RNA via RNA interference (RNAi) , wherein : a . each strand of said siNA molecule is about 18 to about 38 nucleotides in length ; b . one strand of said siNA molecule comprises nucleotide sequence having sufficient complementarity to said target RNA for the siNA molecule to direct cleavage of the target RNA via RNA interference ; and c . wherein one or more nucleotides of said siNA molecule are chemically modified to reduce the immunostimulatory properties of the siNA molecule to a level below that of a corresponding siNA molecule having unmodified nucleotides .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (nucleic acid) mass ratio of from about 5 to about 15 .	US20050282188A1 CLAIM 1 . A chemically synthesized double stranded short interfering nucleic acid (nucleic acid) (siNA) molecule that directs cleavage of a target RNA via RNA interference (RNAi) , wherein : a . each strand of said siNA molecule is about 18 to about 38 nucleotides in length ; b . one strand of said siNA molecule comprises nucleotide sequence having sufficient complementarity to said target RNA for the siNA molecule to direct cleavage of the target RNA via RNA interference ; and c . wherein one or more nucleotides of said siNA molecule are chemically modified to reduce the immunostimulatory properties of the siNA molecule to a level below that of a corresponding siNA molecule having unmodified nucleotides .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof (inflammatory cytokines) comprises from 32 mol % to 36 mol % of the total lipid present in the particle .	US20050282188A1 CLAIM 33 . The method of claim 32 , wherein said reduced immunostimulatory properties comprise an abrogated or reduced induction of inflammatory or proinflammatory cytokines (derivative thereof) in response to the siNA being introduced in a cell , tissue , or organism .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US20050118253A1 Filed: 2004-09-29 Issued: 2005-06-02 Systemic delivery of serum stable plasmid lipid particles for cancer therapy (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Protiva Biotherapeutics Inc Ian Maclachlan, Roger Graham
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US20050118253A1 CLAIM 1 . A method of treating a tumor in a mammal , said method comprising delivering to said tumor a serum-stable nucleic acid-lipid particle comprising a nucleic acid portion that is fully encapsulated within the lipid portion , wherein said delivering is by injection at an injection site that is distal to said tumor in said mammal ; wherein said lipid portion of said nucleic-acid lipid particle comprises a cationic lipid (cationic lipid) , a neutral lipid , and a lipid conjugate that prevents aggregation during formulation : wherein cells of said tumor are responsive to said nucleic acid ; and wherein cells of said tumor are transfectable by said nucleic acid .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	US20050118253A1 CLAIM 1 . A method of treating a tumor in a mammal , said method comprising delivering to said tumor a serum-stable nucleic acid-lipid particle comprising a nucleic acid portion that is fully encapsulated within the lipid portion , wherein said delivering is by injection at an injection site that is distal to said tumor in said mammal ; wherein said lipid portion of said nucleic-acid lipid particle comprises a cationic lipid (cationic lipid) , a neutral lipid , and a lipid conjugate that prevents aggregation during formulation : wherein cells of said tumor are responsive to said nucleic acid ; and wherein cells of said tumor are transfectable by said nucleic acid .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (neutral lipid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	US20050118253A1 CLAIM 1 . A method of treating a tumor in a mammal , said method comprising delivering to said tumor a serum-stable nucleic acid-lipid particle comprising a nucleic acid portion that is fully encapsulated within the lipid portion , wherein said delivering is by injection at an injection site that is distal to said tumor in said mammal ; wherein said lipid portion of said nucleic-acid lipid particle comprises a cationic lipid , a neutral lipid (phospholipid comprises dipalmitoylphosphatidylcholine) , and a lipid conjugate that prevents aggregation during formulation : wherein cells of said tumor are responsive to said nucleic acid ; and wherein cells of said tumor are transfectable by said nucleic acid .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	US20050118253A1 CLAIM 1 . A method of treating a tumor in a mammal , said method comprising delivering to said tumor a serum-stable nucleic acid-lipid particle comprising a nucleic acid portion that is fully encapsulated within the lipid portion , wherein said delivering is by injection at an injection site that is distal to said tumor in said mammal ; wherein said lipid portion of said nucleic-acid lipid particle comprises a cationic lipid (cationic lipid) , a neutral lipid , and a lipid conjugate that prevents aggregation during formulation : wherein cells of said tumor are responsive to said nucleic acid ; and wherein cells of said tumor are transfectable by said nucleic acid .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	WO2005026372A1 Filed: 2004-09-15 Issued: 2005-03-24 Polyethyleneglycol-modified lipid compounds and uses thereof (Original Assignee) Protiva Biotherapeutics, Inc. James Heyes, Ian Maclachlan, Ellen Grace Ambegia
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present (total lipid present) in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	WO2005026372A1 CLAIM 31 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle . WO2005026372A1 CLAIM 48 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said nucleic acid (nucleic acid) is DNA .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	WO2005026372A1 CLAIM 48 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said nucleic acid (nucleic acid) is DNA .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 52 mol % to 62 mol % of the total lipid present (total lipid present) in the particle .	WO2005026372A1 CLAIM 31 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (neutral lipid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	WO2005026372A1 CLAIM 29 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said non-cationic lipid is a neutral lipid (phospholipid comprises dipalmitoylphosphatidylcholine) .
US8058069B2 CLAIM 13 . The nucleic acid-lipid particle of claim 12 , wherein the PEG has an average molecular weight (average molecular weight) of about 2 , 000 daltons .	WO2005026372A1 CLAIM 9 . The compound in accordance with claim 1 , wherein said PEG has an average molecular weight (average molecular weight) ranging from about 550 daltons to about 10 , 000 daltons .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid present (total lipid present) in the particle .	WO2005026372A1 CLAIM 31 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	WO2005026372A1 CLAIM 48 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said nucleic acid (nucleic acid) is DNA .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	WO2005026372A1 CLAIM 48 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said nucleic acid (nucleic acid) is DNA .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	WO2005026372A1 CLAIM 48 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said nucleic acid (nucleic acid) is DNA .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present (total lipid present) in the particle .	WO2005026372A1 CLAIM 31 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid present (total lipid present) in the particle .	WO2005026372A1 CLAIM 31 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US20050064595A1 Filed: 2004-07-16 Issued: 2005-03-24 Lipid encapsulated interfering RNA (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Protiva Biotherapeutics Inc Ian Maclachlan, Ellen Ambegia, James Heyes
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US20050064595A1 CLAIM 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : a siRNA ; a cationic lipid ; a non-cationic lipid ; and a conjugated lipid that inhibits aggregation of particles .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	US20050064595A1 CLAIM 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : a siRNA ; a cationic lipid ; a non-cationic lipid ; and a conjugated lipid that inhibits aggregation of particles .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US20050064595A1 CLAIM 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : a siRNA ; a cationic lipid ; a non-cationic lipid ; and a conjugated lipid that inhibits aggregation of particles .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US20050064595A1 CLAIM 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : a siRNA ; a cationic lipid ; a non-cationic lipid ; and a conjugated lipid that inhibits aggregation of particles .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	US20050064595A1 CLAIM 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : a siRNA ; a cationic lipid ; a non-cationic lipid ; and a conjugated lipid that inhibits aggregation of particles .
US8058069B2 CLAIM 19 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a median diameter (median diameter) of from about 40 nm to about 150 nm .	US20050064595A1 CLAIM 3 . The nucleic acid-lipid particle of claim 1 , wherein said particle has a median diameter (median diameter, lipid particle) of less than about 150 nm .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US20040142892A1 Filed: 2003-10-16 Issued: 2004-07-22 Autogene nucleic acids encoding a secretable RNA polymerase (Original Assignee) University of British Columbia (Current Assignee) University of British Columbia John Finn, Ian Maclachlan
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present (total lipid present) in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US20040142892A1 CLAIM 12 . A lipid-nucleic acid composition comprising : a nucleic acid-lipid particle comprising a lipid portion and a nucleic acid portion , wherein said nucleic acid (nucleic acid) portion comprises an expression cassette comprising two components : (a) a eukaryotic promoter and a first RNA polymerase promoter operably linked to a nucleic acid encoding a secretable RNA polymerase having a secretion domain , and a first internal ribosome entry site ; and (b) a second RNA polymerase promoter operably linked to a nucleic acid encoding a product of interest and a second internal ribosome entry site . US20040142892A1 CLAIM 14 . The lipid-nucleic acid composition of claim 12 , wherein said lipid portion comprises a cationic lipid (cationic lipid) , a non-cationic lipid ; and a polyethyleneglycol-lipid conjugate . US20040142892A1 CLAIM 17 . The lipid-nucleic acid composition of claim 14 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	US20040142892A1 CLAIM 12 . A lipid-nucleic acid composition comprising : a nucleic acid-lipid particle comprising a lipid portion and a nucleic acid portion , wherein said nucleic acid (nucleic acid) portion comprises an expression cassette comprising two components : (a) a eukaryotic promoter and a first RNA polymerase promoter operably linked to a nucleic acid encoding a secretable RNA polymerase having a secretion domain , and a first internal ribosome entry site ; and (b) a second RNA polymerase promoter operably linked to a nucleic acid encoding a product of interest and a second internal ribosome entry site .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present (total lipid present) in the particle .	US20040142892A1 CLAIM 14 . The lipid-nucleic acid composition of claim 12 , wherein said lipid portion comprises a cationic lipid (cationic lipid) , a non-cationic lipid ; and a polyethyleneglycol-lipid conjugate . US20040142892A1 CLAIM 17 . The lipid-nucleic acid composition of claim 14 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	US20040142892A1 CLAIM 14 . The lipid-nucleic acid composition of claim 12 , wherein said lipid portion comprises a cationic lipid (cationic lipid) , a non-cationic lipid ; and a polyethyleneglycol-lipid conjugate .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid present (total lipid present) in the particle .	US20040142892A1 CLAIM 17 . The lipid-nucleic acid composition of claim 14 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US20040142892A1 CLAIM 12 . A lipid-nucleic acid composition comprising : a nucleic acid-lipid particle comprising a lipid portion and a nucleic acid portion , wherein said nucleic acid (nucleic acid) portion comprises an expression cassette comprising two components : (a) a eukaryotic promoter and a first RNA polymerase promoter operably linked to a nucleic acid encoding a secretable RNA polymerase having a secretion domain , and a first internal ribosome entry site ; and (b) a second RNA polymerase promoter operably linked to a nucleic acid encoding a product of interest and a second internal ribosome entry site .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US20040142892A1 CLAIM 12 . A lipid-nucleic acid composition comprising : a nucleic acid-lipid particle comprising a lipid portion and a nucleic acid portion , wherein said nucleic acid (nucleic acid) portion comprises an expression cassette comprising two components : (a) a eukaryotic promoter and a first RNA polymerase promoter operably linked to a nucleic acid encoding a secretable RNA polymerase having a secretion domain , and a first internal ribosome entry site ; and (b) a second RNA polymerase promoter operably linked to a nucleic acid encoding a product of interest and a second internal ribosome entry site .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	US20040142892A1 CLAIM 12 . A lipid-nucleic acid composition comprising : a nucleic acid-lipid particle comprising a lipid portion and a nucleic acid portion , wherein said nucleic acid (nucleic acid) portion comprises an expression cassette comprising two components : (a) a eukaryotic promoter and a first RNA polymerase promoter operably linked to a nucleic acid encoding a secretable RNA polymerase having a secretion domain , and a first internal ribosome entry site ; and (b) a second RNA polymerase promoter operably linked to a nucleic acid encoding a product of interest and a second internal ribosome entry site .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present (total lipid present) in the particle .	US20040142892A1 CLAIM 17 . The lipid-nucleic acid composition of claim 14 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid present (total lipid present) in the particle .	US20040142892A1 CLAIM 17 . The lipid-nucleic acid composition of claim 14 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US20040063654A1 Filed: 2003-05-15 Issued: 2004-04-01 Methods and compositions for therapeutic use of RNA interference (Original Assignee) Insert Therapeutics Inc (Current Assignee) Insert Therapeutics Inc Mark Davis, Gregory Jensen, Suzie Pun
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US20040063654A1 CLAIM 18 . The formulation of claim 17 , wherein the RNAi construct includes a backbone modification selected from phosphorothioates , phosphoramidate , phosphodithioates , chimeric methylphosphonate-phosphodiesters , peptide nucleic acid (nucleic acid) s , and 5-propynyl-pyrimidine containing oligomers .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) comprises a small interfering RNA (siRNA) .	US20040063654A1 CLAIM 18 . The formulation of claim 17 , wherein the RNAi construct includes a backbone modification selected from phosphorothioates , phosphoramidate , phosphodithioates , chimeric methylphosphonate-phosphodiesters , peptide nucleic acid (nucleic acid) s , and 5-propynyl-pyrimidine containing oligomers .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US20040063654A1 CLAIM 18 . The formulation of claim 17 , wherein the RNAi construct includes a backbone modification selected from phosphorothioates , phosphoramidate , phosphodithioates , chimeric methylphosphonate-phosphodiesters , peptide nucleic acid (nucleic acid) s , and 5-propynyl-pyrimidine containing oligomers .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US20040063654A1 CLAIM 18 . The formulation of claim 17 , wherein the RNAi construct includes a backbone modification selected from phosphorothioates , phosphoramidate , phosphodithioates , chimeric methylphosphonate-phosphodiesters , peptide nucleic acid (nucleic acid) s , and 5-propynyl-pyrimidine containing oligomers .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (nucleic acid) mass ratio of from about 5 to about 15 .	US20040063654A1 CLAIM 18 . The formulation of claim 17 , wherein the RNAi construct includes a backbone modification selected from phosphorothioates , phosphoramidate , phosphodithioates , chimeric methylphosphonate-phosphodiesters , peptide nucleic acid (nucleic acid) s , and 5-propynyl-pyrimidine containing oligomers .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US6815432B2 Filed: 2003-02-24 Issued: 2004-11-09 Methods for encapsulating plasmids in lipid bilayers (Original Assignee) Inex Pharmaceuticals Corp (Current Assignee) University of British Columbia ; Arbutus Biopharma Corp Jeffery J. Wheeler, Michael Hope, Pieter R. Cullis, Marcel B. Bally
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present (total lipid present) in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US6815432B2 CLAIM 5 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid comprises from about 2% to about 55% by weight of the total lipid present (total lipid present) in said particle . US6815432B2 CLAIM 13 . The particle of claim 1 , wherein said nucleic acid (nucleic acid) is DNA .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	US6815432B2 CLAIM 13 . The particle of claim 1 , wherein said nucleic acid (nucleic acid) is DNA .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 52 mol % to 62 mol % of the total lipid present (total lipid present) in the particle .	US6815432B2 CLAIM 5 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid comprises from about 2% to about 55% by weight of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid present (total lipid present) in the particle .	US6815432B2 CLAIM 5 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid comprises from about 2% to about 55% by weight of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US6815432B2 CLAIM 13 . The particle of claim 1 , wherein said nucleic acid (nucleic acid) is DNA .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US6815432B2 CLAIM 13 . The particle of claim 1 , wherein said nucleic acid (nucleic acid) is DNA .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	US6815432B2 CLAIM 13 . The particle of claim 1 , wherein said nucleic acid (nucleic acid) is DNA .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present (total lipid present) in the particle .	US6815432B2 CLAIM 5 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid comprises from about 2% to about 55% by weight of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid present (total lipid present) in the particle .	US6815432B2 CLAIM 5 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid comprises from about 2% to about 55% by weight of the total lipid present (total lipid present) in said particle .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US20030216335A1 Filed: 2002-11-27 Issued: 2003-11-20 Method and reagent for the modulation of female reproductive diseases and conditions (Original Assignee) Jennifer Lockridge; Pamela Pavco; Gilad Gordon Jennifer Lockridge, Pamela Pavco, Gilad Gordon
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US20030216335A1 CLAIM 2 . The method of claim 1 , wherein said nucleic acid (nucleic acid) molecule is an enzymatic nucleic acid molecule . US20030216335A1 CLAIM 23 . The method of claim 22 , wherein said lipid is a cationic lipid (cationic lipid) .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	US20030216335A1 CLAIM 2 . The method of claim 1 , wherein said nucleic acid (nucleic acid) molecule is an enzymatic nucleic acid molecule .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	US20030216335A1 CLAIM 23 . The method of claim 22 , wherein said lipid is a cationic lipid (cationic lipid) .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	US20030216335A1 CLAIM 23 . The method of claim 22 , wherein said lipid is a cationic lipid (cationic lipid) .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US20030216335A1 CLAIM 2 . The method of claim 1 , wherein said nucleic acid (nucleic acid) molecule is an enzymatic nucleic acid molecule .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US20030216335A1 CLAIM 2 . The method of claim 1 , wherein said nucleic acid (nucleic acid) molecule is an enzymatic nucleic acid molecule .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	US20030216335A1 CLAIM 2 . The method of claim 1 , wherein said nucleic acid (nucleic acid) molecule is an enzymatic nucleic acid molecule .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US20030082103A1 Filed: 2002-10-01 Issued: 2003-05-01 Targeted therapeutic lipid constructs having cell surface targets (Original Assignee) Targesome Inc (Current Assignee) Nanovalent Pharmaceuticals Inc Charles Wartchow, John Pease, Zhi Shen
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof (EGF receptors) , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US20030082103A1 CLAIM 21 . The lipid construct of claim 1 , wherein said targeting entity is an antibody , protein , ligand , peptide , or nucleic acid (nucleic acid) . US20030082103A1 CLAIM 25 . The lipid construct of claim 1 , wherein said targeting entity is an antibody against one or more of the VEGF receptors (derivative thereof) .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) comprises a small interfering RNA (siRNA) .	US20030082103A1 CLAIM 21 . The lipid construct of claim 1 , wherein said targeting entity is an antibody , protein , ligand , peptide , or nucleic acid (nucleic acid) .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof (EGF receptors) , and about 1 . 4 mol % PEG-lipid conjugate .	US20030082103A1 CLAIM 25 . The lipid construct of claim 1 , wherein said targeting entity is an antibody against one or more of the VEGF receptors (derivative thereof) .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US20030082103A1 CLAIM 21 . The lipid construct of claim 1 , wherein said targeting entity is an antibody , protein , ligand , peptide , or nucleic acid (nucleic acid) .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US20030082103A1 CLAIM 21 . The lipid construct of claim 1 , wherein said targeting entity is an antibody , protein , ligand , peptide , or nucleic acid (nucleic acid) .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (nucleic acid) mass ratio of from about 5 to about 15 .	US20030082103A1 CLAIM 21 . The lipid construct of claim 1 , wherein said targeting entity is an antibody , protein , ligand , peptide , or nucleic acid (nucleic acid) .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof (EGF receptors) comprises from 32 mol % to 36 mol % of the total lipid present in the particle .	US20030082103A1 CLAIM 25 . The lipid construct of claim 1 , wherein said targeting entity is an antibody against one or more of the VEGF receptors (derivative thereof) .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US20030026831A1 Filed: 2002-04-22 Issued: 2003-02-06 Anionic liposomes for delivery of bioactive agents (Original Assignee) University of Minnesota (Current Assignee) University of Minnesota Aparna Lakkaraju, Janet Dubinsky, Walter Low, Yueh-Erh Rahman
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US20030026831A1 CLAIM 18 . The pharmaceutical composition of claim 1 wherein the bioactive agent is an antiviral agent ; an antibacterial agent ; an antifungal agent ; an antineoplastic agent ; an anti-inflammatory agent ; a radiolabel ; a peptide ; a protein ; an oligonucleotide ; a hormone ; a carbohydrate ; a growth factor ; a cytokine ; a radioopaque compound ; a fluorescent compound ; a mydriatic compound ; a bronchodilator ; a local anesthetic ; a nucleic acid (nucleic acid) sequence ; double or single stranded genetic material , or a fragment thereof ; an analgesic ; an antiparasitic ; an antipsychotic ; an antispasmodic ; an arthritis medication ; a biological ; a bone metabolism regulator ; a calcium channel blocker ; a cardiovascular agent ; a central nervous system stimulant ; a diabetes agent ; a diagnostic ; a fungal medication ; a gastrointestinal agent ; a histamine receptor antagonist ; an immunosuppressive ; a muscle relaxant ; a nausea medication ; a nucleoside analogue ; a parkinsonism drug ; a platelet inhibitor ; a psychotropic ; a respiratory drug ; a sedative ; a urinary anti-infective ; a urinary tract agent ; a vitamin ; a nucleotide ; a signaling molecule ; a fluorescent molecule ; a bioactive lipid ; a neuroactive agent ; an energy substrate ; or a combination thereof .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) comprises a small interfering RNA (siRNA) .	US20030026831A1 CLAIM 18 . The pharmaceutical composition of claim 1 wherein the bioactive agent is an antiviral agent ; an antibacterial agent ; an antifungal agent ; an antineoplastic agent ; an anti-inflammatory agent ; a radiolabel ; a peptide ; a protein ; an oligonucleotide ; a hormone ; a carbohydrate ; a growth factor ; a cytokine ; a radioopaque compound ; a fluorescent compound ; a mydriatic compound ; a bronchodilator ; a local anesthetic ; a nucleic acid (nucleic acid) sequence ; double or single stranded genetic material , or a fragment thereof ; an analgesic ; an antiparasitic ; an antipsychotic ; an antispasmodic ; an arthritis medication ; a biological ; a bone metabolism regulator ; a calcium channel blocker ; a cardiovascular agent ; a central nervous system stimulant ; a diabetes agent ; a diagnostic ; a fungal medication ; a gastrointestinal agent ; a histamine receptor antagonist ; an immunosuppressive ; a muscle relaxant ; a nausea medication ; a nucleoside analogue ; a parkinsonism drug ; a platelet inhibitor ; a psychotropic ; a respiratory drug ; a sedative ; a urinary anti-infective ; a urinary tract agent ; a vitamin ; a nucleotide ; a signaling molecule ; a fluorescent molecule ; a bioactive lipid ; a neuroactive agent ; an energy substrate ; or a combination thereof .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US20030026831A1 CLAIM 18 . The pharmaceutical composition of claim 1 wherein the bioactive agent is an antiviral agent ; an antibacterial agent ; an antifungal agent ; an antineoplastic agent ; an anti-inflammatory agent ; a radiolabel ; a peptide ; a protein ; an oligonucleotide ; a hormone ; a carbohydrate ; a growth factor ; a cytokine ; a radioopaque compound ; a fluorescent compound ; a mydriatic compound ; a bronchodilator ; a local anesthetic ; a nucleic acid (nucleic acid) sequence ; double or single stranded genetic material , or a fragment thereof ; an analgesic ; an antiparasitic ; an antipsychotic ; an antispasmodic ; an arthritis medication ; a biological ; a bone metabolism regulator ; a calcium channel blocker ; a cardiovascular agent ; a central nervous system stimulant ; a diabetes agent ; a diagnostic ; a fungal medication ; a gastrointestinal agent ; a histamine receptor antagonist ; an immunosuppressive ; a muscle relaxant ; a nausea medication ; a nucleoside analogue ; a parkinsonism drug ; a platelet inhibitor ; a psychotropic ; a respiratory drug ; a sedative ; a urinary anti-infective ; a urinary tract agent ; a vitamin ; a nucleotide ; a signaling molecule ; a fluorescent molecule ; a bioactive lipid ; a neuroactive agent ; an energy substrate ; or a combination thereof .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US20030026831A1 CLAIM 18 . The pharmaceutical composition of claim 1 wherein the bioactive agent is an antiviral agent ; an antibacterial agent ; an antifungal agent ; an antineoplastic agent ; an anti-inflammatory agent ; a radiolabel ; a peptide ; a protein ; an oligonucleotide ; a hormone ; a carbohydrate ; a growth factor ; a cytokine ; a radioopaque compound ; a fluorescent compound ; a mydriatic compound ; a bronchodilator ; a local anesthetic ; a nucleic acid (nucleic acid) sequence ; double or single stranded genetic material , or a fragment thereof ; an analgesic ; an antiparasitic ; an antipsychotic ; an antispasmodic ; an arthritis medication ; a biological ; a bone metabolism regulator ; a calcium channel blocker ; a cardiovascular agent ; a central nervous system stimulant ; a diabetes agent ; a diagnostic ; a fungal medication ; a gastrointestinal agent ; a histamine receptor antagonist ; an immunosuppressive ; a muscle relaxant ; a nausea medication ; a nucleoside analogue ; a parkinsonism drug ; a platelet inhibitor ; a psychotropic ; a respiratory drug ; a sedative ; a urinary anti-infective ; a urinary tract agent ; a vitamin ; a nucleotide ; a signaling molecule ; a fluorescent molecule ; a bioactive lipid ; a neuroactive agent ; an energy substrate ; or a combination thereof .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (nucleic acid) mass ratio of from about 5 to about 15 .	US20030026831A1 CLAIM 18 . The pharmaceutical composition of claim 1 wherein the bioactive agent is an antiviral agent ; an antibacterial agent ; an antifungal agent ; an antineoplastic agent ; an anti-inflammatory agent ; a radiolabel ; a peptide ; a protein ; an oligonucleotide ; a hormone ; a carbohydrate ; a growth factor ; a cytokine ; a radioopaque compound ; a fluorescent compound ; a mydriatic compound ; a bronchodilator ; a local anesthetic ; a nucleic acid (nucleic acid) sequence ; double or single stranded genetic material , or a fragment thereof ; an analgesic ; an antiparasitic ; an antipsychotic ; an antispasmodic ; an arthritis medication ; a biological ; a bone metabolism regulator ; a calcium channel blocker ; a cardiovascular agent ; a central nervous system stimulant ; a diabetes agent ; a diagnostic ; a fungal medication ; a gastrointestinal agent ; a histamine receptor antagonist ; an immunosuppressive ; a muscle relaxant ; a nausea medication ; a nucleoside analogue ; a parkinsonism drug ; a platelet inhibitor ; a psychotropic ; a respiratory drug ; a sedative ; a urinary anti-infective ; a urinary tract agent ; a vitamin ; a nucleotide ; a signaling molecule ; a fluorescent molecule ; a bioactive lipid ; a neuroactive agent ; an energy substrate ; or a combination thereof . US20030026831A1 CLAIM 48 . The pharmaceutical composition of claim 1 wherein the molar ratio (nucleic acid mass ratio) of bioactive agent to anionic liposome is about 10 : 1 to about 1 : 1×10 10 .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US20030069173A1 Filed: 2001-07-23 Issued: 2003-04-10 Peptide-enhanced transfections (Original Assignee) Life Technologies Inc (Current Assignee) Life Technologies Corp Pamela Hawley-Nelson, Jianqing Lan, PoJen Shih, Joel Jessee, Kevin Schifferli, Gulilat Gebeyehu, Valentina Ciccarone, Krista Evans
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US20030069173A1 CLAIM 22 . The composition of claim 21 , wherein said nucleic acid (nucleic acid) binding groups comprise at least one polyamine .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	US20030069173A1 CLAIM 22 . The composition of claim 21 , wherein said nucleic acid (nucleic acid) binding groups comprise at least one polyamine .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (neutral lipid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	US20030069173A1 CLAIM 7 . The composition of claim 6 , wherein said transfection agent further comprises one or more neutral lipid (phospholipid comprises dipalmitoylphosphatidylcholine) s .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US20030069173A1 CLAIM 22 . The composition of claim 21 , wherein said nucleic acid (nucleic acid) binding groups comprise at least one polyamine .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US20030069173A1 CLAIM 22 . The composition of claim 21 , wherein said nucleic acid (nucleic acid) binding groups comprise at least one polyamine .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	US20030069173A1 CLAIM 22 . The composition of claim 21 , wherein said nucleic acid (nucleic acid) binding groups comprise at least one polyamine .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US20030072794A1 Filed: 2001-06-08 Issued: 2003-04-17 Encapsulation of plasmid DNA (lipogenes™) and therapeutic agents with nuclear localization signal/fusogenic peptide conjugates into targeted liposome complexes (Original Assignee) Teni Boulikas (Current Assignee) Regulon Inc Teni Boulikas
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present (mole percent) in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof (derivative thereof) , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US20030072794A1 CLAIM 1 . A method for producing micelles with entrapped therapeutic agents , comprising : a) combining an effective amount of a negatively charged therapeutic agent with an effective amount of a cationic lipid (cationic lipid) in a ratio where about 30% to about 90% the negatively charged atoms are neutralized by positive charges on lipid molecules to form an electrostatic micelle complex in about 20% to about 80% ethanol ; and b) combining the micelle complex of step a) with an effective amount of a fusogenic-karyophilic peptide conjugates in a ratio range of about 0 . 0 to about 0 . 3 , thereby producing micelles with entrapped therapeutic agents . US20030072794A1 CLAIM 7 . The method of claim 6 , wherein the anionic lipid is dipalmitoyl phosphatidyl glycerol (DDPG) or a derivative thereof (derivative thereof) . US20030072794A1 CLAIM 27 . The method of claim 26 , wherein the liposomes comprises vesicle-forming lipids and between about 1 to about 7 mole percent (total lipid present) of distearoylphosphatidyl ethanolamine (DSPE) derivatized with an effective amount of polyethyleneglycol .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present (mole percent) in the particle .	US20030072794A1 CLAIM 1 . A method for producing micelles with entrapped therapeutic agents , comprising : a) combining an effective amount of a negatively charged therapeutic agent with an effective amount of a cationic lipid (cationic lipid) in a ratio where about 30% to about 90% the negatively charged atoms are neutralized by positive charges on lipid molecules to form an electrostatic micelle complex in about 20% to about 80% ethanol ; and b) combining the micelle complex of step a) with an effective amount of a fusogenic-karyophilic peptide conjugates in a ratio range of about 0 . 0 to about 0 . 3 , thereby producing micelles with entrapped therapeutic agents . US20030072794A1 CLAIM 27 . The method of claim 26 , wherein the liposomes comprises vesicle-forming lipids and between about 1 to about 7 mole percent (total lipid present) of distearoylphosphatidyl ethanolamine (DSPE) derivatized with an effective amount of polyethyleneglycol .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof (derivative thereof) , and about 1 . 4 mol % PEG-lipid conjugate .	US20030072794A1 CLAIM 1 . A method for producing micelles with entrapped therapeutic agents , comprising : a) combining an effective amount of a negatively charged therapeutic agent with an effective amount of a cationic lipid (cationic lipid) in a ratio where about 30% to about 90% the negatively charged atoms are neutralized by positive charges on lipid molecules to form an electrostatic micelle complex in about 20% to about 80% ethanol ; and b) combining the micelle complex of step a) with an effective amount of a fusogenic-karyophilic peptide conjugates in a ratio range of about 0 . 0 to about 0 . 3 , thereby producing micelles with entrapped therapeutic agents . US20030072794A1 CLAIM 7 . The method of claim 6 , wherein the anionic lipid is dipalmitoyl phosphatidyl glycerol (DDPG) or a derivative thereof (derivative thereof) .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid present (mole percent) in the particle .	US20030072794A1 CLAIM 27 . The method of claim 26 , wherein the liposomes comprises vesicle-forming lipids and between about 1 to about 7 mole percent (total lipid present) of distearoylphosphatidyl ethanolamine (DSPE) derivatized with an effective amount of polyethyleneglycol .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid mass ratio (molar ratio) of from about 5 to about 15 .	US20030072794A1 CLAIM 11 . The method of claim 10 , wherein the cationic lipids are combined with the fusogenic lipid DOPE in a molar ratio (nucleic acid mass ratio) from about 1 : 1 to about 2 : 1 .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present (mole percent) in the particle .	US20030072794A1 CLAIM 27 . The method of claim 26 , wherein the liposomes comprises vesicle-forming lipids and between about 1 to about 7 mole percent (total lipid present) of distearoylphosphatidyl ethanolamine (DSPE) derivatized with an effective amount of polyethyleneglycol .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof (derivative thereof) comprises from 32 mol % to 36 mol % of the total lipid present (mole percent) in the particle .	US20030072794A1 CLAIM 7 . The method of claim 6 , wherein the anionic lipid is dipalmitoyl phosphatidyl glycerol (DDPG) or a derivative thereof (derivative thereof) . US20030072794A1 CLAIM 27 . The method of claim 26 , wherein the liposomes comprises vesicle-forming lipids and between about 1 to about 7 mole percent (total lipid present) of distearoylphosphatidyl ethanolamine (DSPE) derivatized with an effective amount of polyethyleneglycol .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US6858224B2 Filed: 2001-06-05 Issued: 2005-02-22 Method of preventing aggregation of a lipid:nucleic acid complex (Original Assignee) Inex Pharmaceuticals Corp (Current Assignee) University of British Columbia ; Arbutus Biopharma Corp Jeffrey Wheeler, Marcel B. Bally, Yuan-Peng Zhang, Dorothy L. Reimer, Michael Hope
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US6858224B2 CLAIM 1 . A method of preventing particle aggregation of lipid : nucleic acid complex particles , said method comprising the step of incorporating an non-cationic lipid (cationic lipid) into lipid : nucleic acid complex particles comprising a cationic lipid and a nucleic acid polymer , wherein said additional lipid is a polyethylene glycol-based polymer . US6858224B2 CLAIM 4 . The method of claim 1 , wherein said nucleic acid (nucleic acid) is selected from the group consisting of DNA and RNA .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	US6858224B2 CLAIM 4 . The method of claim 1 , wherein said nucleic acid (nucleic acid) is selected from the group consisting of DNA and RNA .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	US6858224B2 CLAIM 1 . A method of preventing particle aggregation of lipid : nucleic acid complex particles , said method comprising the step of incorporating an non-cationic lipid (cationic lipid) into lipid : nucleic acid complex particles comprising a cationic lipid and a nucleic acid polymer , wherein said additional lipid is a polyethylene glycol-based polymer .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	US6858224B2 CLAIM 1 . A method of preventing particle aggregation of lipid : nucleic acid complex particles , said method comprising the step of incorporating an non-cationic lipid (cationic lipid) into lipid : nucleic acid complex particles comprising a cationic lipid and a nucleic acid polymer , wherein said additional lipid is a polyethylene glycol-based polymer .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US6858224B2 CLAIM 4 . The method of claim 1 , wherein said nucleic acid (nucleic acid) is selected from the group consisting of DNA and RNA .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US6858224B2 CLAIM 4 . The method of claim 1 , wherein said nucleic acid (nucleic acid) is selected from the group consisting of DNA and RNA .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	US6858224B2 CLAIM 4 . The method of claim 1 , wherein said nucleic acid (nucleic acid) is selected from the group consisting of DNA and RNA .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US6696424B1 Filed: 2000-05-26 Issued: 2004-02-24 Cytofectin dimers and methods of use thereof (Original Assignee) Vical Inc (Current Assignee) Vical Inc Carl Wheeler
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof (derivative thereof) , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US6696424B1 CLAIM 1 . A cationic lipid (cationic lipid) compound of the following formula wherein Z 1 , Z 2 , Z 3 and Z 4 are the same or different and are —O—C(O)— or —O— ; R 1 and R 2 are the same or different and are H , C 1 to C 24 alkyl or C 1 to C 24 alkenyl ; R 3 and R 4 are the same or different and are C 1 to C 24 alkyl or C 1 to C 24 alkenyl ; R 5 , R 6 , R 7 and R 8 are the same or different and are H , C 1 to C 10 alkyl or C 1 to C 10 alkenyl ; R 9 is a linker ; n and m are the same or different and are 1 to 8 ; and X and Y are the same or different and are non-toxic anions ; provided that when R 9 is a straight-chain alkylene having 3-6 , 12 , 16 , 20 , or 22 carbons , then all of R 1 , R 2 , R 3 , and R 4 are not H , all of R 5 , R 6 , R 7 , and R 8 are not methyl , m and n are not 1 , and all of Z 1 , Z 2 , Z 3 , and Z 4 are not O . US6696424B1 CLAIM 21 . The compound of claim 20 , wherein said polyamine is spermine , spermidine , or a derivative thereof (derivative thereof) .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	US6696424B1 CLAIM 1 . A cationic lipid (cationic lipid) compound of the following formula wherein Z 1 , Z 2 , Z 3 and Z 4 are the same or different and are —O—C(O)— or —O— ; R 1 and R 2 are the same or different and are H , C 1 to C 24 alkyl or C 1 to C 24 alkenyl ; R 3 and R 4 are the same or different and are C 1 to C 24 alkyl or C 1 to C 24 alkenyl ; R 5 , R 6 , R 7 and R 8 are the same or different and are H , C 1 to C 10 alkyl or C 1 to C 10 alkenyl ; R 9 is a linker ; n and m are the same or different and are 1 to 8 ; and X and Y are the same or different and are non-toxic anions ; provided that when R 9 is a straight-chain alkylene having 3-6 , 12 , 16 , 20 , or 22 carbons , then all of R 1 , R 2 , R 3 , and R 4 are not H , all of R 5 , R 6 , R 7 , and R 8 are not methyl , m and n are not 1 , and all of Z 1 , Z 2 , Z 3 , and Z 4 are not O .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof (derivative thereof) , and about 1 . 4 mol % PEG-lipid conjugate .	US6696424B1 CLAIM 1 . A cationic lipid (cationic lipid) compound of the following formula wherein Z 1 , Z 2 , Z 3 and Z 4 are the same or different and are —O—C(O)— or —O— ; R 1 and R 2 are the same or different and are H , C 1 to C 24 alkyl or C 1 to C 24 alkenyl ; R 3 and R 4 are the same or different and are C 1 to C 24 alkyl or C 1 to C 24 alkenyl ; R 5 , R 6 , R 7 and R 8 are the same or different and are H , C 1 to C 10 alkyl or C 1 to C 10 alkenyl ; R 9 is a linker ; n and m are the same or different and are 1 to 8 ; and X and Y are the same or different and are non-toxic anions ; provided that when R 9 is a straight-chain alkylene having 3-6 , 12 , 16 , 20 , or 22 carbons , then all of R 1 , R 2 , R 3 , and R 4 are not H , all of R 5 , R 6 , R 7 , and R 8 are not methyl , m and n are not 1 , and all of Z 1 , Z 2 , Z 3 , and Z 4 are not O . US6696424B1 CLAIM 21 . The compound of claim 20 , wherein said polyamine is spermine , spermidine , or a derivative thereof (derivative thereof) .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof (derivative thereof) comprises from 32 mol % to 36 mol % of the total lipid present in the particle .	US6696424B1 CLAIM 21 . The compound of claim 20 , wherein said polyamine is spermine , spermidine , or a derivative thereof (derivative thereof) .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US6586410B1 Filed: 2000-05-08 Issued: 2003-07-01 Lipid-nucleic acid particles prepared via a hydrophobic lipid-nucleic acid complex intermediate and use for gene transfer (Original Assignee) Inex Pharmaceuticals Corp (Current Assignee) University of British Columbia ; Arbutus Biopharma Corp Jeffery J. Wheeler, Marcel B. Bally, Yuan-Peng Zhang, Dorothy L. Reimer, Michael Hope, Pieter R. Cullis, Peter Scherrer
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present (total lipid present) in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US6586410B1 CLAIM 1 . A method of introducing a nucleic acid into a cell , said method comprising contacting said cell with a nucleic acid-lipid particle comprising a cationic lipid (cationic lipid) , a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles , and a nucleic acid , wherein said nucleic acid (nucleic acid) in said nucleic acid-lipid particle is resistant in aqueous solution to degradation with a nuclease . US6586410B1 CLAIM 8 . The method of claim 7 , wherein said PEG-lipid comprises from 1% to about 15% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	US6586410B1 CLAIM 1 . A method of introducing a nucleic acid into a cell , said method comprising contacting said cell with a nucleic acid-lipid particle comprising a cationic lipid , a conjugated lipid that inhibits aggregation of particles , and a nucleic acid , wherein said nucleic acid (nucleic acid) in said nucleic acid-lipid particle is resistant in aqueous solution to degradation with a nuclease .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present (total lipid present) in the particle .	US6586410B1 CLAIM 1 . A method of introducing a nucleic acid into a cell , said method comprising contacting said cell with a nucleic acid-lipid particle comprising a cationic lipid (cationic lipid) , a conjugated lipid that inhibits aggregation of particles , and a nucleic acid , wherein said nucleic acid in said nucleic acid-lipid particle is resistant in aqueous solution to degradation with a nuclease . US6586410B1 CLAIM 8 . The method of claim 7 , wherein said PEG-lipid comprises from 1% to about 15% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 10 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation (inhibits aggregation) of particles comprises a polyethyleneglycol (PEG)-lipid conjugate .	US6586410B1 CLAIM 1 . A method of introducing a nucleic acid into a cell , said method comprising contacting said cell with a nucleic acid-lipid particle comprising a cationic lipid , a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles , and a nucleic acid , wherein said nucleic acid in said nucleic acid-lipid particle is resistant in aqueous solution to degradation with a nuclease .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	US6586410B1 CLAIM 1 . A method of introducing a nucleic acid into a cell , said method comprising contacting said cell with a nucleic acid-lipid particle comprising a cationic lipid (cationic lipid) , a conjugated lipid that inhibits aggregation of particles , and a nucleic acid , wherein said nucleic acid in said nucleic acid-lipid particle is resistant in aqueous solution to degradation with a nuclease .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation (inhibits aggregation) of particles comprises from 1 mol % to 2 mol % of the total lipid present (total lipid present) in the particle .	US6586410B1 CLAIM 1 . A method of introducing a nucleic acid into a cell , said method comprising contacting said cell with a nucleic acid-lipid particle comprising a cationic lipid , a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles , and a nucleic acid , wherein said nucleic acid in said nucleic acid-lipid particle is resistant in aqueous solution to degradation with a nuclease . US6586410B1 CLAIM 8 . The method of claim 7 , wherein said PEG-lipid comprises from 1% to about 15% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US6586410B1 CLAIM 1 . A method of introducing a nucleic acid into a cell , said method comprising contacting said cell with a nucleic acid-lipid particle comprising a cationic lipid , a conjugated lipid that inhibits aggregation of particles , and a nucleic acid , wherein said nucleic acid (nucleic acid) in said nucleic acid-lipid particle is resistant in aqueous solution to degradation with a nuclease .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US6586410B1 CLAIM 1 . A method of introducing a nucleic acid into a cell , said method comprising contacting said cell with a nucleic acid-lipid particle comprising a cationic lipid , a conjugated lipid that inhibits aggregation of particles , and a nucleic acid , wherein said nucleic acid (nucleic acid) in said nucleic acid-lipid particle is resistant in aqueous solution to degradation with a nuclease .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	US6586410B1 CLAIM 1 . A method of introducing a nucleic acid into a cell , said method comprising contacting said cell with a nucleic acid-lipid particle comprising a cationic lipid , a conjugated lipid that inhibits aggregation of particles , and a nucleic acid , wherein said nucleic acid (nucleic acid) in said nucleic acid-lipid particle is resistant in aqueous solution to degradation with a nuclease .
US8058069B2 CLAIM 19 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a median diameter (median diameter, lipid particle) of from about 40 nm to about 150 nm .	US6586410B1 CLAIM 1 . A method of introducing a nucleic acid into a cell , said method comprising contacting said cell with a nucleic acid-lipid particle (median diameter, lipid particle) comprising a cationic lipid , a conjugated lipid that inhibits aggregation of particles , and a nucleic acid , wherein said nucleic acid in said nucleic acid-lipid particle is resistant in aqueous solution to degradation with a nuclease . US6586410B1 CLAIM 3 . The method of claim 1 , wherein said particle has a median diameter (median diameter, lipid particle) of less than about 150 nm .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present (total lipid present) in the particle .	US6586410B1 CLAIM 8 . The method of claim 7 , wherein said PEG-lipid comprises from 1% to about 15% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid present (total lipid present) in the particle .	US6586410B1 CLAIM 8 . The method of claim 7 , wherein said PEG-lipid comprises from 1% to about 15% of the total lipid present (total lipid present) in said particle .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US6852334B1 Filed: 2000-04-20 Issued: 2005-02-08 Cationic peg-lipids and methods of use (Original Assignee) University of British Columbia (Current Assignee) University of British Columbia Pieter R. Cullis, Tao Chen, David B. Fenske, Lorne R. Palmer, Kim Wong
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present (mole percent) in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof (derivative thereof) , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US6852334B1 CLAIM 15 . The vesicle according to claim 1 , wherein said bioactive agent comprises a nucleic acid (nucleic acid) . US6852334B1 CLAIM 28 . A method of increasing an intracellular delivery of a bioactive agent , said method comprising administering to the cell the cationic lipid (cationic lipid) forming vesicle of claim 1 , thereby increasing the intracellular delivery of the bioactive agent . US6852334B1 CLAIM 31 . A method of increasing delivery to a target cell of a drug which is part of a parenterally administered lipid-based drug formulation , said method comprising : preparing a suspension of a lipid-based drug formulation comprising the cationic lipid forming vesicle of claim 1 , wherein between 0 . 1 to 20 mole percent (total lipid present) of said cationic-polymer-lipid conjugated is incorporated . US6852334B1 CLAIM 35 . The vesicle according to claim 1 , wherein Y comprises at least one basic amino acid or derivative thereof (derivative thereof) .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) comprises a small interfering RNA (siRNA) .	US6852334B1 CLAIM 15 . The vesicle according to claim 1 , wherein said bioactive agent comprises a nucleic acid (nucleic acid) .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present (mole percent) in the particle .	US6852334B1 CLAIM 28 . A method of increasing an intracellular delivery of a bioactive agent , said method comprising administering to the cell the cationic lipid (cationic lipid) forming vesicle of claim 1 , thereby increasing the intracellular delivery of the bioactive agent . US6852334B1 CLAIM 31 . A method of increasing delivery to a target cell of a drug which is part of a parenterally administered lipid-based drug formulation , said method comprising : preparing a suspension of a lipid-based drug formulation comprising the cationic lipid forming vesicle of claim 1 , wherein between 0 . 1 to 20 mole percent (total lipid present) of said cationic-polymer-lipid conjugated is incorporated .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof (derivative thereof) , and about 1 . 4 mol % PEG-lipid conjugate .	US6852334B1 CLAIM 28 . A method of increasing an intracellular delivery of a bioactive agent , said method comprising administering to the cell the cationic lipid (cationic lipid) forming vesicle of claim 1 , thereby increasing the intracellular delivery of the bioactive agent . US6852334B1 CLAIM 35 . The vesicle according to claim 1 , wherein Y comprises at least one basic amino acid or derivative thereof (derivative thereof) .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid present (mole percent) in the particle .	US6852334B1 CLAIM 31 . A method of increasing delivery to a target cell of a drug which is part of a parenterally administered lipid-based drug formulation , said method comprising : preparing a suspension of a lipid-based drug formulation comprising the cationic lipid forming vesicle of claim 1 , wherein between 0 . 1 to 20 mole percent (total lipid present) of said cationic-polymer-lipid conjugated is incorporated .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US6852334B1 CLAIM 15 . The vesicle according to claim 1 , wherein said bioactive agent comprises a nucleic acid (nucleic acid) .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US6852334B1 CLAIM 15 . The vesicle according to claim 1 , wherein said bioactive agent comprises a nucleic acid (nucleic acid) .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (nucleic acid) mass ratio of from about 5 to about 15 .	US6852334B1 CLAIM 15 . The vesicle according to claim 1 , wherein said bioactive agent comprises a nucleic acid (nucleic acid) .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present (mole percent) in the particle .	US6852334B1 CLAIM 31 . A method of increasing delivery to a target cell of a drug which is part of a parenterally administered lipid-based drug formulation , said method comprising : preparing a suspension of a lipid-based drug formulation comprising the cationic lipid forming vesicle of claim 1 , wherein between 0 . 1 to 20 mole percent (total lipid present) of said cationic-polymer-lipid conjugated is incorporated .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof (derivative thereof) comprises from 32 mol % to 36 mol % of the total lipid present (mole percent) in the particle .	US6852334B1 CLAIM 31 . A method of increasing delivery to a target cell of a drug which is part of a parenterally administered lipid-based drug formulation , said method comprising : preparing a suspension of a lipid-based drug formulation comprising the cationic lipid forming vesicle of claim 1 , wherein between 0 . 1 to 20 mole percent (total lipid present) of said cationic-polymer-lipid conjugated is incorporated . US6852334B1 CLAIM 35 . The vesicle according to claim 1 , wherein Y comprises at least one basic amino acid or derivative thereof (derivative thereof) .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US7166745B1 Filed: 1999-11-12 Issued: 2007-01-23 Transfection reagents (Original Assignee) Invitrogen Corp (Current Assignee) Life Technologies Corp Yongliang Chu, Malek Masoud, Gulilat Gebeyehu
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US7166745B1 CLAIM 75 . A composition comprising one or more compounds of any one of claims 2 , 5 , 9 , 17 , 20 , 21 , 23 , 25 , 27 , 30 , 32 , 38 , 46 , 52 , 58 , 65 , or 73 and at least one additional component selected from the group consisting of a cell , cells , a cell culture , a cell culture media , a neutral lipid , a nucleic acid (nucleic acid) , and a transfection enhancer . US7166745B1 CLAIM 84 . A composition comprising one or more compounds of any one of claims 2 , 5 , 9 , 17 , 20 , 21 , 23 , 25 , 27 , 30 , 32 , 38 , 46 , 52 , 58 , 65 , or 73 and at least one neutral lipid or at least one other cationic lipid (cationic lipid) .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) comprises a small interfering RNA (siRNA) .	US7166745B1 CLAIM 75 . A composition comprising one or more compounds of any one of claims 2 , 5 , 9 , 17 , 20 , 21 , 23 , 25 , 27 , 30 , 32 , 38 , 46 , 52 , 58 , 65 , or 73 and at least one additional component selected from the group consisting of a cell , cells , a cell culture , a cell culture media , a neutral lipid , a nucleic acid (nucleic acid) , and a transfection enhancer .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	US7166745B1 CLAIM 84 . A composition comprising one or more compounds of any one of claims 2 , 5 , 9 , 17 , 20 , 21 , 23 , 25 , 27 , 30 , 32 , 38 , 46 , 52 , 58 , 65 , or 73 and at least one neutral lipid or at least one other cationic lipid (cationic lipid) .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (neutral lipid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	US7166745B1 CLAIM 75 . A composition comprising one or more compounds of any one of claims 2 , 5 , 9 , 17 , 20 , 21 , 23 , 25 , 27 , 30 , 32 , 38 , 46 , 52 , 58 , 65 , or 73 and at least one additional component selected from the group consisting of a cell , cells , a cell culture , a cell culture media , a neutral lipid (phospholipid comprises dipalmitoylphosphatidylcholine) , a nucleic acid , and a transfection enhancer .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	US7166745B1 CLAIM 84 . A composition comprising one or more compounds of any one of claims 2 , 5 , 9 , 17 , 20 , 21 , 23 , 25 , 27 , 30 , 32 , 38 , 46 , 52 , 58 , 65 , or 73 and at least one neutral lipid or at least one other cationic lipid (cationic lipid) .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US7166745B1 CLAIM 75 . A composition comprising one or more compounds of any one of claims 2 , 5 , 9 , 17 , 20 , 21 , 23 , 25 , 27 , 30 , 32 , 38 , 46 , 52 , 58 , 65 , or 73 and at least one additional component selected from the group consisting of a cell , cells , a cell culture , a cell culture media , a neutral lipid , a nucleic acid (nucleic acid) , and a transfection enhancer .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US7166745B1 CLAIM 75 . A composition comprising one or more compounds of any one of claims 2 , 5 , 9 , 17 , 20 , 21 , 23 , 25 , 27 , 30 , 32 , 38 , 46 , 52 , 58 , 65 , or 73 and at least one additional component selected from the group consisting of a cell , cells , a cell culture , a cell culture media , a neutral lipid , a nucleic acid (nucleic acid) , and a transfection enhancer .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (nucleic acid) mass ratio of from about 5 to about 15 .	US7166745B1 CLAIM 75 . A composition comprising one or more compounds of any one of claims 2 , 5 , 9 , 17 , 20 , 21 , 23 , 25 , 27 , 30 , 32 , 38 , 46 , 52 , 58 , 65 , or 73 and at least one additional component selected from the group consisting of a cell , cells , a cell culture , a cell culture media , a neutral lipid , a nucleic acid (nucleic acid) , and a transfection enhancer .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US6534484B1 Filed: 1999-11-08 Issued: 2003-03-18 Methods for encapsulating plasmids in lipid bilayers (Original Assignee) Inex Pharmaceuticals Corp (Current Assignee) University of British Columbia ; Arbutus Biopharma Corp Jeffery J. Wheeler, Michael Hope, Pieter R. Cullis, Marcel B. Bally
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US6534484B1 CLAIM 1 . A nucleic acid-lipid particle for introducing a nucleic acid into a cell , said particle comprising a cationic lipid (cationic lipid) , a non-cationic lipid , and a nucleic acid , wherein said nucleic acid (nucleic acid) in said nucleic acid-lipid particle is resistant in aqueous solution to degradation by a nuclease .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	US6534484B1 CLAIM 1 . A nucleic acid-lipid particle for introducing a nucleic acid into a cell , said particle comprising a cationic lipid , a non-cationic lipid , and a nucleic acid , wherein said nucleic acid (nucleic acid) in said nucleic acid-lipid particle is resistant in aqueous solution to degradation by a nuclease .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	US6534484B1 CLAIM 1 . A nucleic acid-lipid particle for introducing a nucleic acid into a cell , said particle comprising a cationic lipid (cationic lipid) , a non-cationic lipid , and a nucleic acid , wherein said nucleic acid in said nucleic acid-lipid particle is resistant in aqueous solution to degradation by a nuclease .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	US6534484B1 CLAIM 1 . A nucleic acid-lipid particle for introducing a nucleic acid into a cell , said particle comprising a cationic lipid (cationic lipid) , a non-cationic lipid , and a nucleic acid , wherein said nucleic acid in said nucleic acid-lipid particle is resistant in aqueous solution to degradation by a nuclease .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US6534484B1 CLAIM 1 . A nucleic acid-lipid particle for introducing a nucleic acid into a cell , said particle comprising a cationic lipid , a non-cationic lipid , and a nucleic acid , wherein said nucleic acid (nucleic acid) in said nucleic acid-lipid particle is resistant in aqueous solution to degradation by a nuclease .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US6534484B1 CLAIM 1 . A nucleic acid-lipid particle for introducing a nucleic acid into a cell , said particle comprising a cationic lipid , a non-cationic lipid , and a nucleic acid , wherein said nucleic acid (nucleic acid) in said nucleic acid-lipid particle is resistant in aqueous solution to degradation by a nuclease .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	US6534484B1 CLAIM 1 . A nucleic acid-lipid particle for introducing a nucleic acid into a cell , said particle comprising a cationic lipid , a non-cationic lipid , and a nucleic acid , wherein said nucleic acid (nucleic acid) in said nucleic acid-lipid particle is resistant in aqueous solution to degradation by a nuclease .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US6406705B1 Filed: 1999-06-03 Issued: 2002-06-18 Use of nucleic acids containing unmethylated CpG dinucleotide as an adjuvant (Original Assignee) Ottawa Hospital Research Institute; Coley Pharmaceutical GmbH; University of Iowa Research Foundation UIRF (Current Assignee) Ottawa Hospital Research Institute ; Coley Pharmaceutical GmbH ; University of Iowa Research Foundation UIRF Heather L. Davis, Joachim Schorr, Arthur M. Krieg
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid (unmethylated CpG) present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US6406705B1 CLAIM 1 . A composition of a synergistic combination of adjuvants , comprising : an effective amount for inducing a synergistic adjuvant response of a combination of adjuvants , wherein the combination of adjuvants includes at least one oligonucleotide containing at least one unmethylated CpG (total lipid) dinucleotide and at least one non-nucleic acid (nucleic acid) adjuvant .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) comprises a small interfering RNA (siRNA) .	US6406705B1 CLAIM 1 . A composition of a synergistic combination of adjuvants , comprising : an effective amount for inducing a synergistic adjuvant response of a combination of adjuvants , wherein the combination of adjuvants includes at least one oligonucleotide containing at least one unmethylated CpG dinucleotide and at least one non-nucleic acid (nucleic acid) adjuvant .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 52 mol % to 62 mol % of the total lipid (unmethylated CpG) present in the particle .	US6406705B1 CLAIM 1 . A composition of a synergistic combination of adjuvants , comprising : an effective amount for inducing a synergistic adjuvant response of a combination of adjuvants , wherein the combination of adjuvants includes at least one oligonucleotide containing at least one unmethylated CpG (total lipid) dinucleotide and at least one non-nucleic acid adjuvant .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid (unmethylated CpG) present in the particle .	US6406705B1 CLAIM 1 . A composition of a synergistic combination of adjuvants , comprising : an effective amount for inducing a synergistic adjuvant response of a combination of adjuvants , wherein the combination of adjuvants includes at least one oligonucleotide containing at least one unmethylated CpG (total lipid) dinucleotide and at least one non-nucleic acid adjuvant .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US6406705B1 CLAIM 1 . A composition of a synergistic combination of adjuvants , comprising : an effective amount for inducing a synergistic adjuvant response of a combination of adjuvants , wherein the combination of adjuvants includes at least one oligonucleotide containing at least one unmethylated CpG dinucleotide and at least one non-nucleic acid (nucleic acid) adjuvant .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US6406705B1 CLAIM 1 . A composition of a synergistic combination of adjuvants , comprising : an effective amount for inducing a synergistic adjuvant response of a combination of adjuvants , wherein the combination of adjuvants includes at least one oligonucleotide containing at least one unmethylated CpG dinucleotide and at least one non-nucleic acid (nucleic acid) adjuvant .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (nucleic acid) mass ratio of from about 5 to about 15 .	US6406705B1 CLAIM 1 . A composition of a synergistic combination of adjuvants , comprising : an effective amount for inducing a synergistic adjuvant response of a combination of adjuvants , wherein the combination of adjuvants includes at least one oligonucleotide containing at least one unmethylated CpG dinucleotide and at least one non-nucleic acid (nucleic acid) adjuvant .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid (unmethylated CpG) present in the particle .	US6406705B1 CLAIM 1 . A composition of a synergistic combination of adjuvants , comprising : an effective amount for inducing a synergistic adjuvant response of a combination of adjuvants , wherein the combination of adjuvants includes at least one oligonucleotide containing at least one unmethylated CpG (total lipid) dinucleotide and at least one non-nucleic acid adjuvant .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid (unmethylated CpG) present in the particle .	US6406705B1 CLAIM 1 . A composition of a synergistic combination of adjuvants , comprising : an effective amount for inducing a synergistic adjuvant response of a combination of adjuvants , wherein the combination of adjuvants includes at least one oligonucleotide containing at least one unmethylated CpG (total lipid) dinucleotide and at least one non-nucleic acid adjuvant .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	CA2271582A1 Filed: 1999-05-13 Issued: 1999-11-14 Method for administration of therapeutic agents, including antisense, with repeat dosing (Original Assignee) Sean C. Semple; Sandra K. Klimuk; Troy Harasym; Michael J. Hope; Steven M. Ansell; Pieter R. Cullis; Peter Scherrer; Wilson W. K. Mok; Inex Pharmaceuticals Corp.; University Of British Columbia (Current Assignee) University of British Columbia Sean C. Semple, Sandra K. Klimuk, Troy Harasym, Michael J. Hope, Steven M. Ansell, Pieter R. Cullis, Peter Scherrer, Wilson W. K. Mok
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (lipid component, neutral lipid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	CA2271582A1 CLAIM 1 . A method for the treatment or prevention of a disease characterized by aberrant expression of a gene in a mammalian subject comprising , preparing a lipid-encapsulated therapeutic nucleic acid particle comprising a therapeutic nucleic acid component that hybridizes specifically with the aberrantly expressed gene to reduce its expression in the subject encapsulated within a lipid particle , said lipid particle being formed from a lipid mixture including a steric barrier lipid component (phospholipid comprises dipalmitoylphosphatidylcholine) selected from among lipids that prevent particle aggregation during lipid-nucleic acid particle formation and which exchange out of the lipid particle at a rate greater than PEG-CerC20 ; and administering a therapeutically effective or prophylactic amount of the particle to the mammalian subject in a plurality of separate doses separated in time by intervals of up to eight weeks . CA2271582A1 CLAIM 17 . The method of claim 1 , wherein the lipid mixture comprises an amino lipid having a pKa of from about 5 to about 11 , a neutral lipid (phospholipid comprises dipalmitoylphosphatidylcholine) , Chol and a PEG-modified or polyamide oligomer-modified lipid .

US8058069B2

Filed: 2008-04-15 Issued: 2011-11-15

Lipid formulations for nucleic acid delivery

(Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp

Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan

CA2271582A1

Filed: 1999-05-13 Issued: 1999-11-14

Method for administration of therapeutic agents, including antisense, with repeat dosing

(Original Assignee) Sean C. Semple; Sandra K. Klimuk; Troy Harasym; Michael J. Hope; Steven M. Ansell; Pieter R. Cullis; Peter Scherrer; Wilson W. K. Mok; Inex Pharmaceuticals Corp.; University Of British Columbia (Current Assignee) University of British Columbia

Sean C. Semple, Sandra K. Klimuk, Troy Harasym, Michael J. Hope, Steven M. Ansell, Pieter R. Cullis, Peter Scherrer, Wilson W. K. Mok

US8058069B2
CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (lipid component, neutral lipid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .

CA2271582A1
CLAIM 1 . A method for the treatment or prevention of a disease characterized by aberrant expression of a gene in a mammalian subject comprising , preparing a lipid-encapsulated therapeutic nucleic acid particle comprising a therapeutic nucleic acid component that hybridizes specifically with the aberrantly expressed gene to reduce its expression in the subject encapsulated within a lipid particle , said lipid particle being formed from a lipid mixture including a steric barrier lipid component (phospholipid comprises dipalmitoylphosphatidylcholine) selected from among lipids that prevent particle aggregation during lipid-nucleic acid particle formation and which exchange out of the lipid particle at a rate greater than PEG-CerC20 ;
and administering a therapeutically effective or prophylactic amount of the particle to the mammalian subject in a plurality of separate doses separated in time by intervals of up to eight weeks .

CA2271582A1
CLAIM 17 . The method of claim 1 , wherein the lipid mixture comprises an amino lipid having a pKa of from about 5 to about 11 , a neutral lipid (phospholipid comprises dipalmitoylphosphatidylcholine) , Chol and a PEG-modified or polyamide oligomer-modified lipid .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US6110745A Filed: 1998-07-23 Issued: 2000-08-29 Preparation of lipid-nucleic acid particles using a solvent extraction and direct hydration method (Original Assignee) Inex Pharmaceuticals Corp (Current Assignee) Arbutus Biopharma Corp Yuan-Peng Zhang, Peter Scherrer, Michael J. Hope
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present (mole percent) in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US6110745A CLAIM 1 . A method for the preparation of a lipid-nucloic acid particle , said method comprising : (a) contacting a nucleic acid with a solution comprising a non-cationic lipid (cationic lipid) and a cationic lipid to form a lipid-nucleic acid mixture , said solution comprising about 15-35% water and about 65-85% of an organic solvent ; (b) removing the aqueous portion of said lipid-nucleic acid mixture to form a non-aqueous lipid-nucleic acid mixture ; (c) removing the organic solvent portion from said non-aqueous lipid-nucleic acid mixture to form a lipid-nucleic acid complex in the form of a film ; and (d) hydrating said lipid-nucleic acid complex to form said nucleic acid (nucleic acid) -lipid particle . US6110745A CLAIM 27 . The method in accordance with claim 15 , wherein said PEG-lipid conjugate is present at a concentration ranging from about 0 . 5 mole percent (total lipid present) to about 50 mole percent .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	US6110745A CLAIM 1 . A method for the preparation of a lipid-nucloic acid particle , said method comprising : (a) contacting a nucleic acid with a solution comprising a non-cationic lipid and a cationic lipid to form a lipid-nucleic acid mixture , said solution comprising about 15-35% water and about 65-85% of an organic solvent ; (b) removing the aqueous portion of said lipid-nucleic acid mixture to form a non-aqueous lipid-nucleic acid mixture ; (c) removing the organic solvent portion from said non-aqueous lipid-nucleic acid mixture to form a lipid-nucleic acid complex in the form of a film ; and (d) hydrating said lipid-nucleic acid complex to form said nucleic acid (nucleic acid) -lipid particle .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present (mole percent) in the particle .	US6110745A CLAIM 1 . A method for the preparation of a lipid-nucloic acid particle , said method comprising : (a) contacting a nucleic acid with a solution comprising a non-cationic lipid (cationic lipid) and a cationic lipid to form a lipid-nucleic acid mixture , said solution comprising about 15-35% water and about 65-85% of an organic solvent ; (b) removing the aqueous portion of said lipid-nucleic acid mixture to form a non-aqueous lipid-nucleic acid mixture ; (c) removing the organic solvent portion from said non-aqueous lipid-nucleic acid mixture to form a lipid-nucleic acid complex in the form of a film ; and (d) hydrating said lipid-nucleic acid complex to form said nucleic acid-lipid particle . US6110745A CLAIM 27 . The method in accordance with claim 15 , wherein said PEG-lipid conjugate is present at a concentration ranging from about 0 . 5 mole percent (total lipid present) to about 50 mole percent .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (neutral lipid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	US6110745A CLAIM 5 . The method in accordance with claim 1 , wherein said non-cationic lipid is a neutral lipid (phospholipid comprises dipalmitoylphosphatidylcholine) .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	US6110745A CLAIM 1 . A method for the preparation of a lipid-nucloic acid particle , said method comprising : (a) contacting a nucleic acid with a solution comprising a non-cationic lipid (cationic lipid) and a cationic lipid to form a lipid-nucleic acid mixture , said solution comprising about 15-35% water and about 65-85% of an organic solvent ; (b) removing the aqueous portion of said lipid-nucleic acid mixture to form a non-aqueous lipid-nucleic acid mixture ; (c) removing the organic solvent portion from said non-aqueous lipid-nucleic acid mixture to form a lipid-nucleic acid complex in the form of a film ; and (d) hydrating said lipid-nucleic acid complex to form said nucleic acid-lipid particle .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid present (mole percent) in the particle .	US6110745A CLAIM 27 . The method in accordance with claim 15 , wherein said PEG-lipid conjugate is present at a concentration ranging from about 0 . 5 mole percent (total lipid present) to about 50 mole percent .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US6110745A CLAIM 1 . A method for the preparation of a lipid-nucloic acid particle , said method comprising : (a) contacting a nucleic acid with a solution comprising a non-cationic lipid and a cationic lipid to form a lipid-nucleic acid mixture , said solution comprising about 15-35% water and about 65-85% of an organic solvent ; (b) removing the aqueous portion of said lipid-nucleic acid mixture to form a non-aqueous lipid-nucleic acid mixture ; (c) removing the organic solvent portion from said non-aqueous lipid-nucleic acid mixture to form a lipid-nucleic acid complex in the form of a film ; and (d) hydrating said lipid-nucleic acid complex to form said nucleic acid (nucleic acid) -lipid particle .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US6110745A CLAIM 1 . A method for the preparation of a lipid-nucloic acid particle , said method comprising : (a) contacting a nucleic acid with a solution comprising a non-cationic lipid and a cationic lipid to form a lipid-nucleic acid mixture , said solution comprising about 15-35% water and about 65-85% of an organic solvent ; (b) removing the aqueous portion of said lipid-nucleic acid mixture to form a non-aqueous lipid-nucleic acid mixture ; (c) removing the organic solvent portion from said non-aqueous lipid-nucleic acid mixture to form a lipid-nucleic acid complex in the form of a film ; and (d) hydrating said lipid-nucleic acid complex to form said nucleic acid (nucleic acid) -lipid particle .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	US6110745A CLAIM 1 . A method for the preparation of a lipid-nucloic acid particle , said method comprising : (a) contacting a nucleic acid with a solution comprising a non-cationic lipid and a cationic lipid to form a lipid-nucleic acid mixture , said solution comprising about 15-35% water and about 65-85% of an organic solvent ; (b) removing the aqueous portion of said lipid-nucleic acid mixture to form a non-aqueous lipid-nucleic acid mixture ; (c) removing the organic solvent portion from said non-aqueous lipid-nucleic acid mixture to form a lipid-nucleic acid complex in the form of a film ; and (d) hydrating said lipid-nucleic acid complex to form said nucleic acid (nucleic acid) -lipid particle .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present (mole percent) in the particle .	US6110745A CLAIM 27 . The method in accordance with claim 15 , wherein said PEG-lipid conjugate is present at a concentration ranging from about 0 . 5 mole percent (total lipid present) to about 50 mole percent .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid present (mole percent) in the particle .	US6110745A CLAIM 27 . The method in accordance with claim 15 , wherein said PEG-lipid conjugate is present at a concentration ranging from about 0 . 5 mole percent (total lipid present) to about 50 mole percent .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US6284267B1 Filed: 1997-08-13 Issued: 2001-09-04 Amphiphilic materials and liposome formulations thereof (Original Assignee) Nutrimed Biotech (Current Assignee) Nutrimed Biotech Rajindra Aneja
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US6284267B1 CLAIM 15 . The amphiphilic molecule of claim 1 , wherein at least one of said hydrophobic moieties is selected from the group consisting of an alkylether , a perfluoro analog of an alkylether , a fattyester , a perfluoro analog of a fattyester , a dialkylglycerol , a perfluoro analog of a dialkylglycerol , an alkylamino , a perfluoro analog of an alkylamino , a dialkylamino , a perfluoro analog of a dialkylamino , a diacylglycerol , a perfluoro analog of a diacylglycerol , a sphingolipid , a perfluoro analog of a sphingolipid , a sterol , a perfluoro analog of a sterol , a phospholipid , a perfluoro analog of a phospholipid , a cholestanic acid derivative , a perfluoro analog of a cholestanic acid derivative , a derivative of a synthetic cationic lipid (cationic lipid) precursor and a perfluoro analog of a derivative of a synthetic cationic lipid precursor . US6284267B1 CLAIM 48 . The amphiphilic molecule of claim 43 , wherein said liposome further comprises a selected pharmacological agent , an oxygen carrier , a nutrient , a coagulant , a nucleic acid (nucleic acid) molecule , a nucleic acid vector , an antisense nucleic acid , a ribozyme , a contrast agent or a pheromone .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) comprises a small interfering RNA (siRNA) .	US6284267B1 CLAIM 48 . The amphiphilic molecule of claim 43 , wherein said liposome further comprises a selected pharmacological agent , an oxygen carrier , a nutrient , a coagulant , a nucleic acid (nucleic acid) molecule , a nucleic acid vector , an antisense nucleic acid , a ribozyme , a contrast agent or a pheromone .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	US6284267B1 CLAIM 15 . The amphiphilic molecule of claim 1 , wherein at least one of said hydrophobic moieties is selected from the group consisting of an alkylether , a perfluoro analog of an alkylether , a fattyester , a perfluoro analog of a fattyester , a dialkylglycerol , a perfluoro analog of a dialkylglycerol , an alkylamino , a perfluoro analog of an alkylamino , a dialkylamino , a perfluoro analog of a dialkylamino , a diacylglycerol , a perfluoro analog of a diacylglycerol , a sphingolipid , a perfluoro analog of a sphingolipid , a sterol , a perfluoro analog of a sterol , a phospholipid , a perfluoro analog of a phospholipid , a cholestanic acid derivative , a perfluoro analog of a cholestanic acid derivative , a derivative of a synthetic cationic lipid (cationic lipid) precursor and a perfluoro analog of a derivative of a synthetic cationic lipid precursor .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (lipid component) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	US6284267B1 CLAIM 43 . The amphiphilic molecule of claim 42 , wherein said amphiphilic molecule is formulated with at least one or more lipid component (phospholipid comprises dipalmitoylphosphatidylcholine) s to form a liposome or lipid complex with at least one liposome bilayer .
US8058069B2 CLAIM 13 . The nucleic acid-lipid particle of claim 12 , wherein the PEG has an average molecular weight (average molecular weight) of about 2 , 000 daltons .	US6284267B1 CLAIM 9 . The amphiphilic molecule of claim 8 , wherein said hydrophilic component is a polyethylene glycol with an average molecular weight (average molecular weight) of between about 100 and about 100 , 000 daltons .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	US6284267B1 CLAIM 15 . The amphiphilic molecule of claim 1 , wherein at least one of said hydrophobic moieties is selected from the group consisting of an alkylether , a perfluoro analog of an alkylether , a fattyester , a perfluoro analog of a fattyester , a dialkylglycerol , a perfluoro analog of a dialkylglycerol , an alkylamino , a perfluoro analog of an alkylamino , a dialkylamino , a perfluoro analog of a dialkylamino , a diacylglycerol , a perfluoro analog of a diacylglycerol , a sphingolipid , a perfluoro analog of a sphingolipid , a sterol , a perfluoro analog of a sterol , a phospholipid , a perfluoro analog of a phospholipid , a cholestanic acid derivative , a perfluoro analog of a cholestanic acid derivative , a derivative of a synthetic cationic lipid (cationic lipid) precursor and a perfluoro analog of a derivative of a synthetic cationic lipid precursor .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US6284267B1 CLAIM 48 . The amphiphilic molecule of claim 43 , wherein said liposome further comprises a selected pharmacological agent , an oxygen carrier , a nutrient , a coagulant , a nucleic acid (nucleic acid) molecule , a nucleic acid vector , an antisense nucleic acid , a ribozyme , a contrast agent or a pheromone .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US6284267B1 CLAIM 48 . The amphiphilic molecule of claim 43 , wherein said liposome further comprises a selected pharmacological agent , an oxygen carrier , a nutrient , a coagulant , a nucleic acid (nucleic acid) molecule , a nucleic acid vector , an antisense nucleic acid , a ribozyme , a contrast agent or a pheromone .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (nucleic acid) mass ratio of from about 5 to about 15 .	US6284267B1 CLAIM 48 . The amphiphilic molecule of claim 43 , wherein said liposome further comprises a selected pharmacological agent , an oxygen carrier , a nutrient , a coagulant , a nucleic acid (nucleic acid) molecule , a nucleic acid vector , an antisense nucleic acid , a ribozyme , a contrast agent or a pheromone .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US5976567A Filed: 1996-06-06 Issued: 1999-11-02 Lipid-nucleic acid particles prepared via a hydrophobic lipid-nucleic acid complex intermediate and use for gene transfer (Original Assignee) Inex Pharmaceuticals Corp (Current Assignee) University of British Columbia ; Arbutus Biopharma Corp Jeffery J. Wheeler, Marcel B. Bally, Yuan-Peng Zhang, Dorothy L. Reimer, Michael Hope, Pieter R. Cullis, Peter Scherrer
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US5976567A CLAIM 1 . A method for the preparation of lipid-nucleic acid particles , comprising : (a) contacting nucleic acids with a solution comprising non-cationic lipids and a detergent to form a nucleic acid-lipid mixture ; (b) contacting cationic lipids with said nucleic acid (nucleic acid) -lipid mixture to neutralize the negative charge of said nucleic acids and form a charge-neutralized mixture comprising detergent , nucleic acids and lipids ; and (c) removing said detergent from said charge-neutralized mixture to provide said lipid-nucleic acid particles in which said nucleic acids are protected from degradation .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	US5976567A CLAIM 1 . A method for the preparation of lipid-nucleic acid particles , comprising : (a) contacting nucleic acids with a solution comprising non-cationic lipids and a detergent to form a nucleic acid-lipid mixture ; (b) contacting cationic lipids with said nucleic acid (nucleic acid) -lipid mixture to neutralize the negative charge of said nucleic acids and form a charge-neutralized mixture comprising detergent , nucleic acids and lipids ; and (c) removing said detergent from said charge-neutralized mixture to provide said lipid-nucleic acid particles in which said nucleic acids are protected from degradation .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US5976567A CLAIM 1 . A method for the preparation of lipid-nucleic acid particles , comprising : (a) contacting nucleic acids with a solution comprising non-cationic lipids and a detergent to form a nucleic acid-lipid mixture ; (b) contacting cationic lipids with said nucleic acid (nucleic acid) -lipid mixture to neutralize the negative charge of said nucleic acids and form a charge-neutralized mixture comprising detergent , nucleic acids and lipids ; and (c) removing said detergent from said charge-neutralized mixture to provide said lipid-nucleic acid particles in which said nucleic acids are protected from degradation .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US5976567A CLAIM 1 . A method for the preparation of lipid-nucleic acid particles , comprising : (a) contacting nucleic acids with a solution comprising non-cationic lipids and a detergent to form a nucleic acid-lipid mixture ; (b) contacting cationic lipids with said nucleic acid (nucleic acid) -lipid mixture to neutralize the negative charge of said nucleic acids and form a charge-neutralized mixture comprising detergent , nucleic acids and lipids ; and (c) removing said detergent from said charge-neutralized mixture to provide said lipid-nucleic acid particles in which said nucleic acids are protected from degradation .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	US5976567A CLAIM 1 . A method for the preparation of lipid-nucleic acid particles , comprising : (a) contacting nucleic acids with a solution comprising non-cationic lipids and a detergent to form a nucleic acid-lipid mixture ; (b) contacting cationic lipids with said nucleic acid (nucleic acid) -lipid mixture to neutralize the negative charge of said nucleic acids and form a charge-neutralized mixture comprising detergent , nucleic acids and lipids ; and (c) removing said detergent from said charge-neutralized mixture to provide said lipid-nucleic acid particles in which said nucleic acids are protected from degradation .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US5705385A Filed: 1995-06-07 Issued: 1998-01-06 Lipid-nucleic acid particles prepared via a hydrophobic lipid-nucleic acid complex intermediate and use for gene transfer (Original Assignee) Inex Pharmaceuticals Corp (Current Assignee) University of British Columbia ; Arbutus Biopharma Corp Marcel B. Bally, Yuan-Peng Zhang, Dorothy L. Reimer, Jeffery J. Wheeler
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US5705385A CLAIM 1 . A hydrophobic lipid-nucleic acid complex consisting essentially of cationic lipid (cationic lipid) s and nucleic acids , which complex binds to TO-PRO-1 , and is charge neutralized and soluble in organic solvents . US5705385A CLAIM 2 . A complex in accordance with claim 1 , wherein said nucleic acid (nucleic acid) is a plasmid .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	US5705385A CLAIM 2 . A complex in accordance with claim 1 , wherein said nucleic acid (nucleic acid) is a plasmid .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	US5705385A CLAIM 1 . A hydrophobic lipid-nucleic acid complex consisting essentially of cationic lipid (cationic lipid) s and nucleic acids , which complex binds to TO-PRO-1 , and is charge neutralized and soluble in organic solvents .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	US5705385A CLAIM 1 . A hydrophobic lipid-nucleic acid complex consisting essentially of cationic lipid (cationic lipid) s and nucleic acids , which complex binds to TO-PRO-1 , and is charge neutralized and soluble in organic solvents .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US5705385A CLAIM 2 . A complex in accordance with claim 1 , wherein said nucleic acid (nucleic acid) is a plasmid .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US5705385A CLAIM 2 . A complex in accordance with claim 1 , wherein said nucleic acid (nucleic acid) is a plasmid .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	US5705385A CLAIM 2 . A complex in accordance with claim 1 , wherein said nucleic acid (nucleic acid) is a plasmid .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US5756353A Filed: 1995-06-07 Issued: 1998-05-26 Expression of cloned genes in the lung by aerosol-and liposome-based delivery (Original Assignee) University of California (Current Assignee) University of California Robert J. Debs
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid (particle diameter) present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US5756353A CLAIM 14 . The method of claim 1 , wherein the cationic lipid-DNA complex in aerosol form has a mean particle diameter (total lipid) of less than about 15 μm . US5756353A CLAIM 17 . The method of claim 1 , wherein the cationic lipid-DNA complex comprises a nucleic acid (nucleic acid) base analog .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) comprises a small interfering RNA (siRNA) .	US5756353A CLAIM 17 . The method of claim 1 , wherein the cationic lipid-DNA complex comprises a nucleic acid (nucleic acid) base analog .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 52 mol % to 62 mol % of the total lipid (particle diameter) present in the particle .	US5756353A CLAIM 14 . The method of claim 1 , wherein the cationic lipid-DNA complex in aerosol form has a mean particle diameter (total lipid) of less than about 15 μm .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (lipid component) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	US5756353A CLAIM 10 . The method of claim 1 , wherein the complex comprises DNA and cationic lipid component (phospholipid comprises dipalmitoylphosphatidylcholine) s in a ratio of about 1 mg DNA : 1 micromole cationic lipid .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid (particle diameter) present in the particle .	US5756353A CLAIM 14 . The method of claim 1 , wherein the cationic lipid-DNA complex in aerosol form has a mean particle diameter (total lipid) of less than about 15 μm .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US5756353A CLAIM 17 . The method of claim 1 , wherein the cationic lipid-DNA complex comprises a nucleic acid (nucleic acid) base analog .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US5756353A CLAIM 17 . The method of claim 1 , wherein the cationic lipid-DNA complex comprises a nucleic acid (nucleic acid) base analog .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (nucleic acid) mass ratio of from about 5 to about 15 .	US5756353A CLAIM 17 . The method of claim 1 , wherein the cationic lipid-DNA complex comprises a nucleic acid (nucleic acid) base analog .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid (particle diameter) present in the particle .	US5756353A CLAIM 14 . The method of claim 1 , wherein the cationic lipid-DNA complex in aerosol form has a mean particle diameter (total lipid) of less than about 15 μm .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid (particle diameter) present in the particle .	US5756353A CLAIM 14 . The method of claim 1 , wherein the cationic lipid-DNA complex in aerosol form has a mean particle diameter (total lipid) of less than about 15 μm .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US5820873A Filed: 1995-06-07 Issued: 1998-10-13 Polyethylene glycol modified ceramide lipids and liposome uses thereof (Original Assignee) University of British Columbia (Current Assignee) OF BRITISH COLUMBIA THE, University of ; University of British Columbia Lewis S. L. Choi, Thomas D. Madden, Murray S. Webb
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present (mole percent) in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US5820873A CLAIM 25 . The liposome of claim 24 , wherein mole percent (total lipid present) ratio of the lipid is about 0 . 01 to about 60 .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 52 mol % to 62 mol % of the total lipid present (mole percent) in the particle .	US5820873A CLAIM 25 . The liposome of claim 24 , wherein mole percent (total lipid present) ratio of the lipid is about 0 . 01 to about 60 .
US8058069B2 CLAIM 13 . The nucleic acid-lipid particle of claim 12 , wherein the PEG has an average molecular weight (average molecular weight) of about 2 , 000 daltons .	US5820873A CLAIM 1 . A lipid compound of the formula ##STR8## wherein : R 1 , R 2 , and R 3 are independently hydrogen , C 1 -C 6 alkyl , acyl , or aryl ; R 4 is hydrogen , C 1 -C 30 alkyl , C 2 -C 30 alkenyl , C 2 -C 30 alkynyl , or aryl ; R 5 is hydrogen , alkyl , acyl , aryl , or PEG ; X 1 is --O-- , --S-- , or --NR 6 -- , where R 6 is hydrogen , C 1 -C 6 alkyl , acyl or aryl ; or when R 5 is PEG and b is 1 , X 1 is also --Y 1 -alk-Y 2 ; Y is --NR 7 -- , where R 7 is hydrogen , C 1 -C 6 alkyl , acyl or aryl , or Y is --O-- , --S-- or --Y 1 -alk-Y 2 -- , wherein Y 1 and Y 2 are independently amino , amido , carboxyl , carbamate , carbonyl , carbonate , urea , or phosphoro ; and alk is C 1 -C 6 alkylene ; PEG is a polyethylene glycol with an average molecular weight (average molecular weight) from about 550 to about 8 , 500 daltons optionally substituted by C 1 -C 3 alkyl , alkoxy , acyl or aryl ; wherein a is 0 or 1 ; and b is 1 unless R 5 is PEG wherein b is 0 or 1 .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid present (mole percent) in the particle .	US5820873A CLAIM 25 . The liposome of claim 24 , wherein mole percent (total lipid present) ratio of the lipid is about 0 . 01 to about 60 .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present (mole percent) in the particle .	US5820873A CLAIM 25 . The liposome of claim 24 , wherein mole percent (total lipid present) ratio of the lipid is about 0 . 01 to about 60 .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid present (mole percent) in the particle .	US5820873A CLAIM 25 . The liposome of claim 24 , wherein mole percent (total lipid present) ratio of the lipid is about 0 . 01 to about 60 .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US5885613A Filed: 1995-06-07 Issued: 1999-03-23 Bilayer stabilizing components and their use in forming programmable fusogenic liposomes (Original Assignee) University of British Columbia (Current Assignee) University of British Columbia John W. Holland, Thomas D. Madden, Pieter R. Cullis
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present (mole percent) in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US5885613A CLAIM 1 . A fusogenic liposome comprising : a lipid capable of adopting a non-lamellar phase , yet capable of assuming a bilayer structure in the presence of a polyethyleneglycol-ceramide conjugate , wherein said lipid is a member selected from the group consisting of phosphatidylenthanolamines , phosphatidylserines , ceramides , glycolipids and mixtures thereof ; and a polyethyleneglycol-ceramide conjugate reversibly associated with said lipid to stabilize said lipid in a bilayer structure , wherein said polyethyleneglycol-ceramide conjugate is present at a concentration ranging from about 0 . 05 mole percent (total lipid present) to about 50 mole percent . US5885613A CLAIM 5 . The fusogenic liposome in accordance with claim 1 wherein said lipid is a mixture of a phosphatidylethanolamine and a cationic lipid (cationic lipid) .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present (mole percent) in the particle .	US5885613A CLAIM 1 . A fusogenic liposome comprising : a lipid capable of adopting a non-lamellar phase , yet capable of assuming a bilayer structure in the presence of a polyethyleneglycol-ceramide conjugate , wherein said lipid is a member selected from the group consisting of phosphatidylenthanolamines , phosphatidylserines , ceramides , glycolipids and mixtures thereof ; and a polyethyleneglycol-ceramide conjugate reversibly associated with said lipid to stabilize said lipid in a bilayer structure , wherein said polyethyleneglycol-ceramide conjugate is present at a concentration ranging from about 0 . 05 mole percent (total lipid present) to about 50 mole percent . US5885613A CLAIM 5 . The fusogenic liposome in accordance with claim 1 wherein said lipid is a mixture of a phosphatidylethanolamine and a cationic lipid (cationic lipid) .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	US5885613A CLAIM 5 . The fusogenic liposome in accordance with claim 1 wherein said lipid is a mixture of a phosphatidylethanolamine and a cationic lipid (cationic lipid) .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid present (mole percent) in the particle .	US5885613A CLAIM 1 . A fusogenic liposome comprising : a lipid capable of adopting a non-lamellar phase , yet capable of assuming a bilayer structure in the presence of a polyethyleneglycol-ceramide conjugate , wherein said lipid is a member selected from the group consisting of phosphatidylenthanolamines , phosphatidylserines , ceramides , glycolipids and mixtures thereof ; and a polyethyleneglycol-ceramide conjugate reversibly associated with said lipid to stabilize said lipid in a bilayer structure , wherein said polyethyleneglycol-ceramide conjugate is present at a concentration ranging from about 0 . 05 mole percent (total lipid present) to about 50 mole percent .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present (mole percent) in the particle .	US5885613A CLAIM 1 . A fusogenic liposome comprising : a lipid capable of adopting a non-lamellar phase , yet capable of assuming a bilayer structure in the presence of a polyethyleneglycol-ceramide conjugate , wherein said lipid is a member selected from the group consisting of phosphatidylenthanolamines , phosphatidylserines , ceramides , glycolipids and mixtures thereof ; and a polyethyleneglycol-ceramide conjugate reversibly associated with said lipid to stabilize said lipid in a bilayer structure , wherein said polyethyleneglycol-ceramide conjugate is present at a concentration ranging from about 0 . 05 mole percent (total lipid present) to about 50 mole percent .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid present (mole percent) in the particle .	US5885613A CLAIM 1 . A fusogenic liposome comprising : a lipid capable of adopting a non-lamellar phase , yet capable of assuming a bilayer structure in the presence of a polyethyleneglycol-ceramide conjugate , wherein said lipid is a member selected from the group consisting of phosphatidylenthanolamines , phosphatidylserines , ceramides , glycolipids and mixtures thereof ; and a polyethyleneglycol-ceramide conjugate reversibly associated with said lipid to stabilize said lipid in a bilayer structure , wherein said polyethyleneglycol-ceramide conjugate is present at a concentration ranging from about 0 . 05 mole percent (total lipid present) to about 50 mole percent .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US5627159A Filed: 1994-10-27 Issued: 1997-05-06 Enhancement of lipid cationic transfections in the presence of serum (Original Assignee) Life Technologies Inc (Current Assignee) Life Technologies Corp PoJen Shih, Pamela Hawley-Nelson, Joel A. Jessee
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US5627159A CLAIM 1 . In a method of transfecting an animal cell in the presence of serum , comprising contacting said cell with a lipid aggregate comprising nucleic acid and a cationic lipid (cationic lipid) , wherein the improvement comprises : contacting said cell with said lipid aggregate in the presence of a polycationic compound , thereby transfecting said animal cell with said nucleic acid (nucleic acid) .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	US5627159A CLAIM 1 . In a method of transfecting an animal cell in the presence of serum , comprising contacting said cell with a lipid aggregate comprising nucleic acid and a cationic lipid , wherein the improvement comprises : contacting said cell with said lipid aggregate in the presence of a polycationic compound , thereby transfecting said animal cell with said nucleic acid (nucleic acid) .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	US5627159A CLAIM 1 . In a method of transfecting an animal cell in the presence of serum , comprising contacting said cell with a lipid aggregate comprising nucleic acid and a cationic lipid (cationic lipid) , wherein the improvement comprises : contacting said cell with said lipid aggregate in the presence of a polycationic compound , thereby transfecting said animal cell with said nucleic acid .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	US5627159A CLAIM 1 . In a method of transfecting an animal cell in the presence of serum , comprising contacting said cell with a lipid aggregate comprising nucleic acid and a cationic lipid (cationic lipid) , wherein the improvement comprises : contacting said cell with said lipid aggregate in the presence of a polycationic compound , thereby transfecting said animal cell with said nucleic acid .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US5627159A CLAIM 1 . In a method of transfecting an animal cell in the presence of serum , comprising contacting said cell with a lipid aggregate comprising nucleic acid and a cationic lipid , wherein the improvement comprises : contacting said cell with said lipid aggregate in the presence of a polycationic compound , thereby transfecting said animal cell with said nucleic acid (nucleic acid) .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US5627159A CLAIM 1 . In a method of transfecting an animal cell in the presence of serum , comprising contacting said cell with a lipid aggregate comprising nucleic acid and a cationic lipid , wherein the improvement comprises : contacting said cell with said lipid aggregate in the presence of a polycationic compound , thereby transfecting said animal cell with said nucleic acid (nucleic acid) .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	US5627159A CLAIM 1 . In a method of transfecting an animal cell in the presence of serum , comprising contacting said cell with a lipid aggregate comprising nucleic acid and a cationic lipid , wherein the improvement comprises : contacting said cell with said lipid aggregate in the presence of a polycationic compound , thereby transfecting said animal cell with said nucleic acid (nucleic acid) .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US5578475A Filed: 1994-07-12 Issued: 1996-11-26 Composition and methods for transfecting eukaryotic cells (Original Assignee) Life Technologies Inc (Current Assignee) Life Technologies Corp Joel A. Jessee
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US5578475A CLAIM 1 . A composition for transfecting a eukaryotic cell wherein said composition comprises a nucleic acid , a cationic lipid composition selected from the group consisting of a 3 : 1 (w/w) mixture of the polycationic lipid , 2 , 3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N , N-dimethyl-1-propanaminiumtrifluoroacetate and dioleoylphosphatidylethanolamine , and a 1 : 1 (w/w) mixture of N-[1-(2 , 3-dioleyloxy)propyl]-N , N , N-trimethylammoniumchloride and dioleoylphosphatidylethanolamine and a viral agent which enhances transfection two-fold or more by said cationic lipid composition , which viral agent is an alphavirus or a component of an alphavirus and wherein said nucleic acid (nucleic acid) is not a nucleic acid of said viral agent .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	US5578475A CLAIM 1 . A composition for transfecting a eukaryotic cell wherein said composition comprises a nucleic acid , a cationic lipid composition selected from the group consisting of a 3 : 1 (w/w) mixture of the polycationic lipid , 2 , 3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N , N-dimethyl-1-propanaminiumtrifluoroacetate and dioleoylphosphatidylethanolamine , and a 1 : 1 (w/w) mixture of N-[1-(2 , 3-dioleyloxy)propyl]-N , N , N-trimethylammoniumchloride and dioleoylphosphatidylethanolamine and a viral agent which enhances transfection two-fold or more by said cationic lipid composition , which viral agent is an alphavirus or a component of an alphavirus and wherein said nucleic acid (nucleic acid) is not a nucleic acid of said viral agent .
US8058069B2 CLAIM 13 . The nucleic acid-lipid particle of claim 12 , wherein the PEG has an average molecular weight (said component) of about 2 , 000 daltons .	US5578475A CLAIM 36 . The transfection composition of claim 1 wherein said component (average molecular weight) of an alphavirus is a spike glycoprotein .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US5578475A CLAIM 1 . A composition for transfecting a eukaryotic cell wherein said composition comprises a nucleic acid , a cationic lipid composition selected from the group consisting of a 3 : 1 (w/w) mixture of the polycationic lipid , 2 , 3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N , N-dimethyl-1-propanaminiumtrifluoroacetate and dioleoylphosphatidylethanolamine , and a 1 : 1 (w/w) mixture of N-[1-(2 , 3-dioleyloxy)propyl]-N , N , N-trimethylammoniumchloride and dioleoylphosphatidylethanolamine and a viral agent which enhances transfection two-fold or more by said cationic lipid composition , which viral agent is an alphavirus or a component of an alphavirus and wherein said nucleic acid (nucleic acid) is not a nucleic acid of said viral agent .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US5578475A CLAIM 1 . A composition for transfecting a eukaryotic cell wherein said composition comprises a nucleic acid , a cationic lipid composition selected from the group consisting of a 3 : 1 (w/w) mixture of the polycationic lipid , 2 , 3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N , N-dimethyl-1-propanaminiumtrifluoroacetate and dioleoylphosphatidylethanolamine , and a 1 : 1 (w/w) mixture of N-[1-(2 , 3-dioleyloxy)propyl]-N , N , N-trimethylammoniumchloride and dioleoylphosphatidylethanolamine and a viral agent which enhances transfection two-fold or more by said cationic lipid composition , which viral agent is an alphavirus or a component of an alphavirus and wherein said nucleic acid (nucleic acid) is not a nucleic acid of said viral agent .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	US5578475A CLAIM 1 . A composition for transfecting a eukaryotic cell wherein said composition comprises a nucleic acid , a cationic lipid composition selected from the group consisting of a 3 : 1 (w/w) mixture of the polycationic lipid , 2 , 3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N , N-dimethyl-1-propanaminiumtrifluoroacetate and dioleoylphosphatidylethanolamine , and a 1 : 1 (w/w) mixture of N-[1-(2 , 3-dioleyloxy)propyl]-N , N , N-trimethylammoniumchloride and dioleoylphosphatidylethanolamine and a viral agent which enhances transfection two-fold or more by said cationic lipid composition , which viral agent is an alphavirus or a component of an alphavirus and wherein said nucleic acid (nucleic acid) is not a nucleic acid of said viral agent .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US5703055A Filed: 1994-01-26 Issued: 1997-12-30 Generation of antibodies through lipid mediated DNA delivery (Original Assignee) Vical Inc; Wisconsin Alumni Research Foundation (Current Assignee) Vical Inc ; Wisconsin Alumni Research Foundation Philip L. Felgner, Jon Asher Wolff, Gary H. Rhodes, Robert Wallace Malone, Dennis A. Carson
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US5703055A CLAIM 1 . A method of generating desired antibodies in a mammal comprising : directly administering to a tissue of a mammal a DNA sequence operatively linked to a promoter or a mRNA sequence encoding an immunogen , said sequence being complexed to a cationic lipid (cationic lipid) , in an amount sufficient to induce the detectable production of desired antibodies to the expressed immunogen .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	US5703055A CLAIM 1 . A method of generating desired antibodies in a mammal comprising : directly administering to a tissue of a mammal a DNA sequence operatively linked to a promoter or a mRNA sequence encoding an immunogen , said sequence being complexed to a cationic lipid (cationic lipid) , in an amount sufficient to induce the detectable production of desired antibodies to the expressed immunogen .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (neutral lipid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	US5703055A CLAIM 10 . The method of claim 1 , wherein said cationic lipid is formulated as a positively charged lipid species with a neutral lipid (phospholipid comprises dipalmitoylphosphatidylcholine) species .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	US5703055A CLAIM 1 . A method of generating desired antibodies in a mammal comprising : directly administering to a tissue of a mammal a DNA sequence operatively linked to a promoter or a mRNA sequence encoding an immunogen , said sequence being complexed to a cationic lipid (cationic lipid) , in an amount sufficient to induce the detectable production of desired antibodies to the expressed immunogen .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US5674908A Filed: 1993-12-20 Issued: 1997-10-07 Highly packed polycationic ammonium, sulfonium and phosphonium lipids (Original Assignee) Life Technologies Inc (Current Assignee) Life Technologies Corp Alberto Haces, Valentina C. Ciccarone
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US5674908A CLAIM 34 . A composition for transfecting a cell with a nucleic acid (nucleic acid) which comprises a nucleic acid and one or more compounds of claim 1 .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) comprises a small interfering RNA (siRNA) .	US5674908A CLAIM 34 . A composition for transfecting a cell with a nucleic acid (nucleic acid) which comprises a nucleic acid and one or more compounds of claim 1 .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (neutral lipid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	US5674908A CLAIM 31 . A lipid aggregate of claim 30 wherein said second compound is a neutral lipid (phospholipid comprises dipalmitoylphosphatidylcholine) .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US5674908A CLAIM 34 . A composition for transfecting a cell with a nucleic acid (nucleic acid) which comprises a nucleic acid and one or more compounds of claim 1 .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US5674908A CLAIM 34 . A composition for transfecting a cell with a nucleic acid (nucleic acid) which comprises a nucleic acid and one or more compounds of claim 1 .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (nucleic acid) mass ratio of from about 5 to about 15 .	US5674908A CLAIM 34 . A composition for transfecting a cell with a nucleic acid (nucleic acid) which comprises a nucleic acid and one or more compounds of claim 1 .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US5320906A Filed: 1991-12-20 Issued: 1994-06-14 Delivery vehicles with amphiphile-associated active ingredient (Original Assignee) Vestar Inc (Current Assignee) NeXstar Pharmaceuticals Inc Crispin G. S. Eley, Paul G. Schmidt, Gary Fujii
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (fatty acids) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US5320906A CLAIM 5 . The delivery vehicle of claim 4 wherein the amphiphilic material comprises a member selected from the group consisting of fatty acids (nucleic acid) , phospholipids , diglycerides , triglycerides , alcohols , amines , phosphates and sulfates .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (fatty acids) comprises a small interfering RNA (siRNA) .	US5320906A CLAIM 5 . The delivery vehicle of claim 4 wherein the amphiphilic material comprises a member selected from the group consisting of fatty acids (nucleic acid) , phospholipids , diglycerides , triglycerides , alcohols , amines , phosphates and sulfates .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (fatty acids) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US5320906A CLAIM 5 . The delivery vehicle of claim 4 wherein the amphiphilic material comprises a member selected from the group consisting of fatty acids (nucleic acid) , phospholipids , diglycerides , triglycerides , alcohols , amines , phosphates and sulfates .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (fatty acids) is fully encapsulated in the nucleic acid-lipid particle .	US5320906A CLAIM 5 . The delivery vehicle of claim 4 wherein the amphiphilic material comprises a member selected from the group consisting of fatty acids (nucleic acid) , phospholipids , diglycerides , triglycerides , alcohols , amines , phosphates and sulfates .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (fatty acids) mass ratio of from about 5 to about 15 .	US5320906A CLAIM 5 . The delivery vehicle of claim 4 wherein the amphiphilic material comprises a member selected from the group consisting of fatty acids (nucleic acid) , phospholipids , diglycerides , triglycerides , alcohols , amines , phosphates and sulfates .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US5283185A Filed: 1991-08-28 Issued: 1994-02-01 Method for delivering nucleic acids into cells (Original Assignee) McMaster University; University of Tennessee Research Foundation (Current Assignee) McMaster University ; University of Tennessee Research Foundation Richard M. Epand, Remo Bottega, Leaf Huang
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US5283185A CLAIM 1 . A method for facilitating the transfer of nucleic acid (nucleic acid) s into mammalian cells with a stable aqueous dispersion of mixed lipids which dispersion comprises : a cationic lipid (cationic lipid) which is a weak protein kinase C inhibitor and has a structure which includes a lipophilic group derived from cholesterol , a linker bond , selected from the group consisting of carboxy amides and carbamoyls , a spacer arm having from 1 to 20 carbon atoms in a linear branched or unbranched alkyl chain , and a cationic amino group selected from the group consisting of primary , secondary , tertiary and quaternary amino groups , and a co-lipid selected from the groups consisting of phosphatidylcholine and phosphatidylethanolamine which method comprises mixing said aqueous dispersion of mixed lipids with the nucleic acids , thereby forming a dispersion/nucleic acids complex , and incubating the cells to be transfected with the complex , thereby facilitating the transfer of the nucleic acids into the cells .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) comprises a small interfering RNA (siRNA) .	US5283185A CLAIM 1 . A method for facilitating the transfer of nucleic acid (nucleic acid) s into mammalian cells with a stable aqueous dispersion of mixed lipids which dispersion comprises : a cationic lipid which is a weak protein kinase C inhibitor and has a structure which includes a lipophilic group derived from cholesterol , a linker bond , selected from the group consisting of carboxy amides and carbamoyls , a spacer arm having from 1 to 20 carbon atoms in a linear branched or unbranched alkyl chain , and a cationic amino group selected from the group consisting of primary , secondary , tertiary and quaternary amino groups , and a co-lipid selected from the groups consisting of phosphatidylcholine and phosphatidylethanolamine which method comprises mixing said aqueous dispersion of mixed lipids with the nucleic acids , thereby forming a dispersion/nucleic acids complex , and incubating the cells to be transfected with the complex , thereby facilitating the transfer of the nucleic acids into the cells .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	US5283185A CLAIM 1 . A method for facilitating the transfer of nucleic acids into mammalian cells with a stable aqueous dispersion of mixed lipids which dispersion comprises : a cationic lipid (cationic lipid) which is a weak protein kinase C inhibitor and has a structure which includes a lipophilic group derived from cholesterol , a linker bond , selected from the group consisting of carboxy amides and carbamoyls , a spacer arm having from 1 to 20 carbon atoms in a linear branched or unbranched alkyl chain , and a cationic amino group selected from the group consisting of primary , secondary , tertiary and quaternary amino groups , and a co-lipid selected from the groups consisting of phosphatidylcholine and phosphatidylethanolamine which method comprises mixing said aqueous dispersion of mixed lipids with the nucleic acids , thereby forming a dispersion/nucleic acids complex , and incubating the cells to be transfected with the complex , thereby facilitating the transfer of the nucleic acids into the cells .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	US5283185A CLAIM 1 . A method for facilitating the transfer of nucleic acids into mammalian cells with a stable aqueous dispersion of mixed lipids which dispersion comprises : a cationic lipid (cationic lipid) which is a weak protein kinase C inhibitor and has a structure which includes a lipophilic group derived from cholesterol , a linker bond , selected from the group consisting of carboxy amides and carbamoyls , a spacer arm having from 1 to 20 carbon atoms in a linear branched or unbranched alkyl chain , and a cationic amino group selected from the group consisting of primary , secondary , tertiary and quaternary amino groups , and a co-lipid selected from the groups consisting of phosphatidylcholine and phosphatidylethanolamine which method comprises mixing said aqueous dispersion of mixed lipids with the nucleic acids , thereby forming a dispersion/nucleic acids complex , and incubating the cells to be transfected with the complex , thereby facilitating the transfer of the nucleic acids into the cells .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US5283185A CLAIM 1 . A method for facilitating the transfer of nucleic acid (nucleic acid) s into mammalian cells with a stable aqueous dispersion of mixed lipids which dispersion comprises : a cationic lipid which is a weak protein kinase C inhibitor and has a structure which includes a lipophilic group derived from cholesterol , a linker bond , selected from the group consisting of carboxy amides and carbamoyls , a spacer arm having from 1 to 20 carbon atoms in a linear branched or unbranched alkyl chain , and a cationic amino group selected from the group consisting of primary , secondary , tertiary and quaternary amino groups , and a co-lipid selected from the groups consisting of phosphatidylcholine and phosphatidylethanolamine which method comprises mixing said aqueous dispersion of mixed lipids with the nucleic acids , thereby forming a dispersion/nucleic acids complex , and incubating the cells to be transfected with the complex , thereby facilitating the transfer of the nucleic acids into the cells .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US5283185A CLAIM 1 . A method for facilitating the transfer of nucleic acid (nucleic acid) s into mammalian cells with a stable aqueous dispersion of mixed lipids which dispersion comprises : a cationic lipid which is a weak protein kinase C inhibitor and has a structure which includes a lipophilic group derived from cholesterol , a linker bond , selected from the group consisting of carboxy amides and carbamoyls , a spacer arm having from 1 to 20 carbon atoms in a linear branched or unbranched alkyl chain , and a cationic amino group selected from the group consisting of primary , secondary , tertiary and quaternary amino groups , and a co-lipid selected from the groups consisting of phosphatidylcholine and phosphatidylethanolamine which method comprises mixing said aqueous dispersion of mixed lipids with the nucleic acids , thereby forming a dispersion/nucleic acids complex , and incubating the cells to be transfected with the complex , thereby facilitating the transfer of the nucleic acids into the cells .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (nucleic acid) mass ratio of from about 5 to about 15 .	US5283185A CLAIM 1 . A method for facilitating the transfer of nucleic acid (nucleic acid) s into mammalian cells with a stable aqueous dispersion of mixed lipids which dispersion comprises : a cationic lipid which is a weak protein kinase C inhibitor and has a structure which includes a lipophilic group derived from cholesterol , a linker bond , selected from the group consisting of carboxy amides and carbamoyls , a spacer arm having from 1 to 20 carbon atoms in a linear branched or unbranched alkyl chain , and a cationic amino group selected from the group consisting of primary , secondary , tertiary and quaternary amino groups , and a co-lipid selected from the groups consisting of phosphatidylcholine and phosphatidylethanolamine which method comprises mixing said aqueous dispersion of mixed lipids with the nucleic acids , thereby forming a dispersion/nucleic acids complex , and incubating the cells to be transfected with the complex , thereby facilitating the transfer of the nucleic acids into the cells .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US5225212A Filed: 1990-12-10 Issued: 1993-07-06 Microreservoir liposome composition and method (Original Assignee) Liposome Technology Inc (Current Assignee) Liposome Technology Inc Francis J. Martin, Martin C. Woodle, Carl Redemann, Annie Yau-Young, Ramachandran Radhakrishnan
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid (particle diameter) present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US5225212A CLAIM 1 . A liposome composition effective to extend , to at least 24 hours , the period of effective activity of a therapeutic compound which can be administered intravenously in a therapeutically effective amount and which is cleared in free form from the bloodstream with a halflife of less than about 4 hours , comprising liposomes (i) composed of vesicle-forming lipids and between 1-20 mole percent (total lipid present) of a vesicle-forming lipid derivatized with a polymer selected from the group consisting of polyethyleneglycol , polyacetic acid and polyglycolic acid , and (ii) having a selected mean particle diameter (total lipid) in the size range between about 0 . 1 to 0 . 4 microns , and the compound in liposome-entrapped form , for intravenous administration at a dose of the composition which contains an amount of the compound in liposome-entrapped form which is at least three times such therapeutically effective amount .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 52 mol % to 62 mol % of the total lipid (particle diameter) present in the particle .	US5225212A CLAIM 1 . A liposome composition effective to extend , to at least 24 hours , the period of effective activity of a therapeutic compound which can be administered intravenously in a therapeutically effective amount and which is cleared in free form from the bloodstream with a halflife of less than about 4 hours , comprising liposomes (i) composed of vesicle-forming lipids and between 1-20 mole percent (total lipid present) of a vesicle-forming lipid derivatized with a polymer selected from the group consisting of polyethyleneglycol , polyacetic acid and polyglycolic acid , and (ii) having a selected mean particle diameter (total lipid) in the size range between about 0 . 1 to 0 . 4 microns , and the compound in liposome-entrapped form , for intravenous administration at a dose of the composition which contains an amount of the compound in liposome-entrapped form which is at least three times such therapeutically effective amount .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid (particle diameter) present in the particle .	US5225212A CLAIM 1 . A liposome composition effective to extend , to at least 24 hours , the period of effective activity of a therapeutic compound which can be administered intravenously in a therapeutically effective amount and which is cleared in free form from the bloodstream with a halflife of less than about 4 hours , comprising liposomes (i) composed of vesicle-forming lipids and between 1-20 mole percent (total lipid present) of a vesicle-forming lipid derivatized with a polymer selected from the group consisting of polyethyleneglycol , polyacetic acid and polyglycolic acid , and (ii) having a selected mean particle diameter (total lipid) in the size range between about 0 . 1 to 0 . 4 microns , and the compound in liposome-entrapped form , for intravenous administration at a dose of the composition which contains an amount of the compound in liposome-entrapped form which is at least three times such therapeutically effective amount .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid (particle diameter) present in the particle .	US5225212A CLAIM 1 . A liposome composition effective to extend , to at least 24 hours , the period of effective activity of a therapeutic compound which can be administered intravenously in a therapeutically effective amount and which is cleared in free form from the bloodstream with a halflife of less than about 4 hours , comprising liposomes (i) composed of vesicle-forming lipids and between 1-20 mole percent (total lipid present) of a vesicle-forming lipid derivatized with a polymer selected from the group consisting of polyethyleneglycol , polyacetic acid and polyglycolic acid , and (ii) having a selected mean particle diameter (total lipid) in the size range between about 0 . 1 to 0 . 4 microns , and the compound in liposome-entrapped form , for intravenous administration at a dose of the composition which contains an amount of the compound in liposome-entrapped form which is at least three times such therapeutically effective amount .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid (particle diameter) present in the particle .	US5225212A CLAIM 1 . A liposome composition effective to extend , to at least 24 hours , the period of effective activity of a therapeutic compound which can be administered intravenously in a therapeutically effective amount and which is cleared in free form from the bloodstream with a halflife of less than about 4 hours , comprising liposomes (i) composed of vesicle-forming lipids and between 1-20 mole percent (total lipid present) of a vesicle-forming lipid derivatized with a polymer selected from the group consisting of polyethyleneglycol , polyacetic acid and polyglycolic acid , and (ii) having a selected mean particle diameter (total lipid) in the size range between about 0 . 1 to 0 . 4 microns , and the compound in liposome-entrapped form , for intravenous administration at a dose of the composition which contains an amount of the compound in liposome-entrapped form which is at least three times such therapeutically effective amount .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US5279833A Filed: 1990-04-04 Issued: 1994-01-18 Liposomal transfection of nucleic acids into animal cells (Original Assignee) Yale University (Current Assignee) YALE UNIVERSITY A NON-PROFIT CT ; Yale University John K. Rose
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US5279833A CLAIM 1 . A liposome for introducing a nucleic acid (nucleic acid) into an animal cell comprising (a) a neutral lipid selected from the group consisting of lecithin , lysolecithin , lysophosphatidylethanolamine , phosphatidylserine , phosphatidylinositol , sphinogomyelin , cephalin , cardiolipin , phosphatidic acid , cerebrosides , dioleoylphosphatidylcholine , dipalmitoylphosphatidylcholine , dioleoylphosphatidylglycerol , dipalmitoylphosphatidylglycerol , palmitoyloleoylphosphatidylcholine , palmitoyloleoylphosphatidylethanolamine , diheptadecanoylphosphatidylethanolamine , dilauroylphosphatidylethanolamine , dimyristoylphosphatidylethanolamine , distearoylphosphatidylethanolamine , beta-linoleoyl-gamma-palmitoylphosphatidylethanolamine and beta-oleoyl-gamma-palmitoylphosphatidylethanolamine , and (b) a cationic lipid (cationic lipid) selection from the group consisting of dimethyldioctadecylammonium bromide , cetylidimethylethylammonium bromide , stearylamine and methylbenzethonium chloride , wherein the weight ratio of the cationic lipid to the neutral lipid is 6 : 30 to 12 : 30 .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) comprises a small interfering RNA (siRNA) .	US5279833A CLAIM 1 . A liposome for introducing a nucleic acid (nucleic acid) into an animal cell comprising (a) a neutral lipid selected from the group consisting of lecithin , lysolecithin , lysophosphatidylethanolamine , phosphatidylserine , phosphatidylinositol , sphinogomyelin , cephalin , cardiolipin , phosphatidic acid , cerebrosides , dioleoylphosphatidylcholine , dipalmitoylphosphatidylcholine , dioleoylphosphatidylglycerol , dipalmitoylphosphatidylglycerol , palmitoyloleoylphosphatidylcholine , palmitoyloleoylphosphatidylethanolamine , diheptadecanoylphosphatidylethanolamine , dilauroylphosphatidylethanolamine , dimyristoylphosphatidylethanolamine , distearoylphosphatidylethanolamine , beta-linoleoyl-gamma-palmitoylphosphatidylethanolamine and beta-oleoyl-gamma-palmitoylphosphatidylethanolamine , and (b) a cationic lipid selection from the group consisting of dimethyldioctadecylammonium bromide , cetylidimethylethylammonium bromide , stearylamine and methylbenzethonium chloride , wherein the weight ratio of the cationic lipid to the neutral lipid is 6 : 30 to 12 : 30 .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	US5279833A CLAIM 1 . A liposome for introducing a nucleic acid into an animal cell comprising (a) a neutral lipid selected from the group consisting of lecithin , lysolecithin , lysophosphatidylethanolamine , phosphatidylserine , phosphatidylinositol , sphinogomyelin , cephalin , cardiolipin , phosphatidic acid , cerebrosides , dioleoylphosphatidylcholine , dipalmitoylphosphatidylcholine , dioleoylphosphatidylglycerol , dipalmitoylphosphatidylglycerol , palmitoyloleoylphosphatidylcholine , palmitoyloleoylphosphatidylethanolamine , diheptadecanoylphosphatidylethanolamine , dilauroylphosphatidylethanolamine , dimyristoylphosphatidylethanolamine , distearoylphosphatidylethanolamine , beta-linoleoyl-gamma-palmitoylphosphatidylethanolamine and beta-oleoyl-gamma-palmitoylphosphatidylethanolamine , and (b) a cationic lipid (cationic lipid) selection from the group consisting of dimethyldioctadecylammonium bromide , cetylidimethylethylammonium bromide , stearylamine and methylbenzethonium chloride , wherein the weight ratio of the cationic lipid to the neutral lipid is 6 : 30 to 12 : 30 .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (neutral lipid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	US5279833A CLAIM 1 . A liposome for introducing a nucleic acid into an animal cell comprising (a) a neutral lipid (phospholipid comprises dipalmitoylphosphatidylcholine) selected from the group consisting of lecithin , lysolecithin , lysophosphatidylethanolamine , phosphatidylserine , phosphatidylinositol , sphinogomyelin , cephalin , cardiolipin , phosphatidic acid , cerebrosides , dioleoylphosphatidylcholine , dipalmitoylphosphatidylcholine , dioleoylphosphatidylglycerol , dipalmitoylphosphatidylglycerol , palmitoyloleoylphosphatidylcholine , palmitoyloleoylphosphatidylethanolamine , diheptadecanoylphosphatidylethanolamine , dilauroylphosphatidylethanolamine , dimyristoylphosphatidylethanolamine , distearoylphosphatidylethanolamine , beta-linoleoyl-gamma-palmitoylphosphatidylethanolamine and beta-oleoyl-gamma-palmitoylphosphatidylethanolamine , and (b) a cationic lipid selection from the group consisting of dimethyldioctadecylammonium bromide , cetylidimethylethylammonium bromide , stearylamine and methylbenzethonium chloride , wherein the weight ratio of the cationic lipid to the neutral lipid is 6 : 30 to 12 : 30 .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	US5279833A CLAIM 1 . A liposome for introducing a nucleic acid into an animal cell comprising (a) a neutral lipid selected from the group consisting of lecithin , lysolecithin , lysophosphatidylethanolamine , phosphatidylserine , phosphatidylinositol , sphinogomyelin , cephalin , cardiolipin , phosphatidic acid , cerebrosides , dioleoylphosphatidylcholine , dipalmitoylphosphatidylcholine , dioleoylphosphatidylglycerol , dipalmitoylphosphatidylglycerol , palmitoyloleoylphosphatidylcholine , palmitoyloleoylphosphatidylethanolamine , diheptadecanoylphosphatidylethanolamine , dilauroylphosphatidylethanolamine , dimyristoylphosphatidylethanolamine , distearoylphosphatidylethanolamine , beta-linoleoyl-gamma-palmitoylphosphatidylethanolamine and beta-oleoyl-gamma-palmitoylphosphatidylethanolamine , and (b) a cationic lipid (cationic lipid) selection from the group consisting of dimethyldioctadecylammonium bromide , cetylidimethylethylammonium bromide , stearylamine and methylbenzethonium chloride , wherein the weight ratio of the cationic lipid to the neutral lipid is 6 : 30 to 12 : 30 .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US5279833A CLAIM 1 . A liposome for introducing a nucleic acid (nucleic acid) into an animal cell comprising (a) a neutral lipid selected from the group consisting of lecithin , lysolecithin , lysophosphatidylethanolamine , phosphatidylserine , phosphatidylinositol , sphinogomyelin , cephalin , cardiolipin , phosphatidic acid , cerebrosides , dioleoylphosphatidylcholine , dipalmitoylphosphatidylcholine , dioleoylphosphatidylglycerol , dipalmitoylphosphatidylglycerol , palmitoyloleoylphosphatidylcholine , palmitoyloleoylphosphatidylethanolamine , diheptadecanoylphosphatidylethanolamine , dilauroylphosphatidylethanolamine , dimyristoylphosphatidylethanolamine , distearoylphosphatidylethanolamine , beta-linoleoyl-gamma-palmitoylphosphatidylethanolamine and beta-oleoyl-gamma-palmitoylphosphatidylethanolamine , and (b) a cationic lipid selection from the group consisting of dimethyldioctadecylammonium bromide , cetylidimethylethylammonium bromide , stearylamine and methylbenzethonium chloride , wherein the weight ratio of the cationic lipid to the neutral lipid is 6 : 30 to 12 : 30 .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US5279833A CLAIM 1 . A liposome for introducing a nucleic acid (nucleic acid) into an animal cell comprising (a) a neutral lipid selected from the group consisting of lecithin , lysolecithin , lysophosphatidylethanolamine , phosphatidylserine , phosphatidylinositol , sphinogomyelin , cephalin , cardiolipin , phosphatidic acid , cerebrosides , dioleoylphosphatidylcholine , dipalmitoylphosphatidylcholine , dioleoylphosphatidylglycerol , dipalmitoylphosphatidylglycerol , palmitoyloleoylphosphatidylcholine , palmitoyloleoylphosphatidylethanolamine , diheptadecanoylphosphatidylethanolamine , dilauroylphosphatidylethanolamine , dimyristoylphosphatidylethanolamine , distearoylphosphatidylethanolamine , beta-linoleoyl-gamma-palmitoylphosphatidylethanolamine and beta-oleoyl-gamma-palmitoylphosphatidylethanolamine , and (b) a cationic lipid selection from the group consisting of dimethyldioctadecylammonium bromide , cetylidimethylethylammonium bromide , stearylamine and methylbenzethonium chloride , wherein the weight ratio of the cationic lipid to the neutral lipid is 6 : 30 to 12 : 30 .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (nucleic acid) mass ratio of from about 5 to about 15 .	US5279833A CLAIM 1 . A liposome for introducing a nucleic acid (nucleic acid) into an animal cell comprising (a) a neutral lipid selected from the group consisting of lecithin , lysolecithin , lysophosphatidylethanolamine , phosphatidylserine , phosphatidylinositol , sphinogomyelin , cephalin , cardiolipin , phosphatidic acid , cerebrosides , dioleoylphosphatidylcholine , dipalmitoylphosphatidylcholine , dioleoylphosphatidylglycerol , dipalmitoylphosphatidylglycerol , palmitoyloleoylphosphatidylcholine , palmitoyloleoylphosphatidylethanolamine , diheptadecanoylphosphatidylethanolamine , dilauroylphosphatidylethanolamine , dimyristoylphosphatidylethanolamine , distearoylphosphatidylethanolamine , beta-linoleoyl-gamma-palmitoylphosphatidylethanolamine and beta-oleoyl-gamma-palmitoylphosphatidylethanolamine , and (b) a cationic lipid selection from the group consisting of dimethyldioctadecylammonium bromide , cetylidimethylethylammonium bromide , stearylamine and methylbenzethonium chloride , wherein the weight ratio of the cationic lipid to the neutral lipid is 6 : 30 to 12 : 30 .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US4987071A Filed: 1986-12-03 Issued: 1991-01-22 RNA ribozyme polymerases, dephosphorylases, restriction endoribonucleases and methods (Original Assignee) University Patents Inc (Current Assignee) University Patents Inc Thomas R. Cech, Arthur J. Zaug, Michael D. Been
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US4987071A CLAIM 14 . The enzymatic RNA molecule of any of claims 1-6 wherein said enzymatic RNA molecule further comprises a second endonuclease activity , independent of any protein , for nucleic acid (nucleic acid) comprising deoxynucleotides .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) comprises a small interfering RNA (siRNA) .	US4987071A CLAIM 14 . The enzymatic RNA molecule of any of claims 1-6 wherein said enzymatic RNA molecule further comprises a second endonuclease activity , independent of any protein , for nucleic acid (nucleic acid) comprising deoxynucleotides .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (polycytidylic acid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	US4987071A CLAIM 17 . The enzymatic RNA molecule of claim 15 wherein said nucleotidyltransferase activity causes polymerization of oligocytidylic acid to polycytidylic acid (phospholipid comprises dipalmitoylphosphatidylcholine) .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US4987071A CLAIM 14 . The enzymatic RNA molecule of any of claims 1-6 wherein said enzymatic RNA molecule further comprises a second endonuclease activity , independent of any protein , for nucleic acid (nucleic acid) comprising deoxynucleotides .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US4987071A CLAIM 14 . The enzymatic RNA molecule of any of claims 1-6 wherein said enzymatic RNA molecule further comprises a second endonuclease activity , independent of any protein , for nucleic acid (nucleic acid) comprising deoxynucleotides .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (nucleic acid) mass ratio of from about 5 to about 15 .	US4987071A CLAIM 14 . The enzymatic RNA molecule of any of claims 1-6 wherein said enzymatic RNA molecule further comprises a second endonuclease activity , independent of any protein , for nucleic acid (nucleic acid) comprising deoxynucleotides .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	WO2007051303A1 Filed: 2006-11-02 Issued: 2007-05-10 MODIFIED siRNA MOLECULES AND USES THEREOF (Original Assignee) Protiva Biotherapeutics, Inc. Ian Maclachlan, Adam Judge
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present (total lipid present) in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	WO2007051303A1 CLAIM 66 . The nucleic acid-lipid particle of claim 59 , further comprising a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles . WO2007051303A1 CLAIM 72 . The nucleic acid-lipid particle of claim 59 , wherein the cationic lipid comprises from about 20 mol % to about 50 mol % of the total lipid present (total lipid present) in the particle .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 52 mol % to 62 mol % of the total lipid present (total lipid present) in the particle .	WO2007051303A1 CLAIM 72 . The nucleic acid-lipid particle of claim 59 , wherein the cationic lipid comprises from about 20 mol % to about 50 mol % of the total lipid present (total lipid present) in the particle .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (neutral lipid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	WO2007051303A1 CLAIM 63 . The nucleic acid-lipid particle of claim 59 , wherein the non-cationic lipid is a neutral lipid (phospholipid comprises dipalmitoylphosphatidylcholine) .
US8058069B2 CLAIM 10 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation (inhibits aggregation) of particles comprises a polyethyleneglycol (PEG)-lipid conjugate .	WO2007051303A1 CLAIM 66 . The nucleic acid-lipid particle of claim 59 , further comprising a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation (inhibits aggregation) of particles comprises from 1 mol % to 2 mol % of the total lipid present (total lipid present) in the particle .	WO2007051303A1 CLAIM 66 . The nucleic acid-lipid particle of claim 59 , further comprising a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles . WO2007051303A1 CLAIM 72 . The nucleic acid-lipid particle of claim 59 , wherein the cationic lipid comprises from about 20 mol % to about 50 mol % of the total lipid present (total lipid present) in the particle .
US8058069B2 CLAIM 19 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a median diameter (median diameter) of from about 40 nm to about 150 nm .	WO2007051303A1 CLAIM 87 . The nucleic acid-lipid particle of claim 59 , wherein the particle has a median diameter (median diameter, lipid particle) of from about 50 nm to about 150 nm .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present (total lipid present) in the particle .	WO2007051303A1 CLAIM 72 . The nucleic acid-lipid particle of claim 59 , wherein the cationic lipid comprises from about 20 mol % to about 50 mol % of the total lipid present (total lipid present) in the particle .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid present (total lipid present) in the particle .	WO2007051303A1 CLAIM 72 . The nucleic acid-lipid particle of claim 59 , wherein the cationic lipid comprises from about 20 mol % to about 50 mol % of the total lipid present (total lipid present) in the particle .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	WO2007024323A2 Filed: 2006-06-19 Issued: 2007-03-01 Nanoparticle fabrication methods, systems, and materials (Original Assignee) The University Of North Carolina At Chapel Hill; Liquidia Technologies Inc.; North Carolina State University Joseph M. Desimone, Jason P. Rolland, Ansley E. Exner, Edward T. Samulski, R. Jude Samulski, Benjamin W. Maynor, Larken E. Euliss, Ginger Denison Rothrock, Stephanie Gratton, Alex Ermosh, Andrew James Murphy
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (carboxylic acids) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid (biodegradable poly, metal precursor) present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	WO2007024323A2 CLAIM 3 . The composition of claim 2 , wherein the biocompatible material is selected from the group consisting of a poly(ethylene glycol) , a poly(lactic acid) , a poly(lactic acid-co-glycolic acid) , a lactose , a phosphatidylcholine , a polylactide , a polyglycolide , a hydroxypropylcellulose , a wax , a polyester , a polyanhydride , a polyamide , a phosphorous-based polymer , a poly(cyanoacrylate) , a polyurethane , a polyorthoester , a polydihydropyran , a polyacetal , a biodegradable poly (lipid particle, total lipid) mer , a polypeptide , a hydrogel , a carbohydrate , and combinations thereof . WO2007024323A2 CLAIM 36 . The composition of claim 4 , wherein the linker is selected from the group consisting of sulfides , amines , carboxylic acids (nucleic acid) , acid chlorides , alcohols , alkenes , alkyl halides , isocyanates , imidazoles , halides , azides , N- hydroxysuccimidyl (NHS) ester groups , acetylenes , diethylenetriaminepentaacetic acid (DPTA) and combinations thereof . WO2007024323A2 CLAIM 142 . The method of claim 110 , wherein the substance to be molded is selected from the group consisting of a polymer , a solution , a monomer , a plurality of monomers , a polymerization initiator , a polymerization catalyst , an inorganic precursor , a metal precursor (lipid particle, total lipid) , a pharmaceutical agent , a tag , a magnetic material , a paramagnetic material , a ligand , a cell penetrating peptide , a porogen , a surfactant , a plurality of immiscible liquids , a solvent , and a charged species .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (carboxylic acids) comprises a small interfering RNA (siRNA) .	WO2007024323A2 CLAIM 36 . The composition of claim 4 , wherein the linker is selected from the group consisting of sulfides , amines , carboxylic acids (nucleic acid) , acid chlorides , alcohols , alkenes , alkyl halides , isocyanates , imidazoles , halides , azides , N- hydroxysuccimidyl (NHS) ester groups , acetylenes , diethylenetriaminepentaacetic acid (DPTA) and combinations thereof .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 52 mol % to 62 mol % of the total lipid (biodegradable poly, metal precursor) present in the particle .	WO2007024323A2 CLAIM 3 . The composition of claim 2 , wherein the biocompatible material is selected from the group consisting of a poly(ethylene glycol) , a poly(lactic acid) , a poly(lactic acid-co-glycolic acid) , a lactose , a phosphatidylcholine , a polylactide , a polyglycolide , a hydroxypropylcellulose , a wax , a polyester , a polyanhydride , a polyamide , a phosphorous-based polymer , a poly(cyanoacrylate) , a polyurethane , a polyorthoester , a polydihydropyran , a polyacetal , a biodegradable poly (lipid particle, total lipid) mer , a polypeptide , a hydrogel , a carbohydrate , and combinations thereof . WO2007024323A2 CLAIM 142 . The method of claim 110 , wherein the substance to be molded is selected from the group consisting of a polymer , a solution , a monomer , a plurality of monomers , a polymerization initiator , a polymerization catalyst , an inorganic precursor , a metal precursor (lipid particle, total lipid) , a pharmaceutical agent , a tag , a magnetic material , a paramagnetic material , a ligand , a cell penetrating peptide , a porogen , a surfactant , a plurality of immiscible liquids , a solvent , and a charged species .
US8058069B2 CLAIM 13 . The nucleic acid-lipid particle of claim 12 , wherein the PEG has an average molecular weight (polymerization catalyst, cellulose ethers) of about 2 , 000 daltons .	WO2007024323A2 CLAIM 64 . The composition of claim 22 , wherein the super absorbent polymer is selected from the group consisting of polyacrylates , polyacrylic acid , HEMA , neutralized acrylates , sodium acrylate , ammonium acrylate , methacrylates , polyacrylamide , cellulose ethers (average molecular weight) , poly (ethylene oxide) , poly (vinyl alcohol) , polysuccinimides , polyacrylonitrile polymers , combinations of the above polymers blended or crosslinked together , combinations of the above polymers having monomers co-polymerized with monomers of another polymer , combinations of the above polymers with starch , and combinations thereof . WO2007024323A2 CLAIM 142 . The method of claim 110 , wherein the substance to be molded is selected from the group consisting of a polymer , a solution , a monomer , a plurality of monomers , a polymerization initiator , a polymerization catalyst (average molecular weight) , an inorganic precursor , a metal precursor , a pharmaceutical agent , a tag , a magnetic material , a paramagnetic material , a ligand , a cell penetrating peptide , a porogen , a surfactant , a plurality of immiscible liquids , a solvent , and a charged species .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid (biodegradable poly, metal precursor) present in the particle .	WO2007024323A2 CLAIM 3 . The composition of claim 2 , wherein the biocompatible material is selected from the group consisting of a poly(ethylene glycol) , a poly(lactic acid) , a poly(lactic acid-co-glycolic acid) , a lactose , a phosphatidylcholine , a polylactide , a polyglycolide , a hydroxypropylcellulose , a wax , a polyester , a polyanhydride , a polyamide , a phosphorous-based polymer , a poly(cyanoacrylate) , a polyurethane , a polyorthoester , a polydihydropyran , a polyacetal , a biodegradable poly (lipid particle, total lipid) mer , a polypeptide , a hydrogel , a carbohydrate , and combinations thereof . WO2007024323A2 CLAIM 142 . The method of claim 110 , wherein the substance to be molded is selected from the group consisting of a polymer , a solution , a monomer , a plurality of monomers , a polymerization initiator , a polymerization catalyst , an inorganic precursor , a metal precursor (lipid particle, total lipid) , a pharmaceutical agent , a tag , a magnetic material , a paramagnetic material , a ligand , a cell penetrating peptide , a porogen , a surfactant , a plurality of immiscible liquids , a solvent , and a charged species .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (carboxylic acids) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	WO2007024323A2 CLAIM 36 . The composition of claim 4 , wherein the linker is selected from the group consisting of sulfides , amines , carboxylic acids (nucleic acid) , acid chlorides , alcohols , alkenes , alkyl halides , isocyanates , imidazoles , halides , azides , N- hydroxysuccimidyl (NHS) ester groups , acetylenes , diethylenetriaminepentaacetic acid (DPTA) and combinations thereof .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (carboxylic acids) is fully encapsulated in the nucleic acid-lipid particle .	WO2007024323A2 CLAIM 36 . The composition of claim 4 , wherein the linker is selected from the group consisting of sulfides , amines , carboxylic acids (nucleic acid) , acid chlorides , alcohols , alkenes , alkyl halides , isocyanates , imidazoles , halides , azides , N- hydroxysuccimidyl (NHS) ester groups , acetylenes , diethylenetriaminepentaacetic acid (DPTA) and combinations thereof .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (carboxylic acids) mass ratio of from about 5 to about 15 .	WO2007024323A2 CLAIM 36 . The composition of claim 4 , wherein the linker is selected from the group consisting of sulfides , amines , carboxylic acids (nucleic acid) , acid chlorides , alcohols , alkenes , alkyl halides , isocyanates , imidazoles , halides , azides , N- hydroxysuccimidyl (NHS) ester groups , acetylenes , diethylenetriaminepentaacetic acid (DPTA) and combinations thereof .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid (biodegradable poly, metal precursor) present in the particle .	WO2007024323A2 CLAIM 3 . The composition of claim 2 , wherein the biocompatible material is selected from the group consisting of a poly(ethylene glycol) , a poly(lactic acid) , a poly(lactic acid-co-glycolic acid) , a lactose , a phosphatidylcholine , a polylactide , a polyglycolide , a hydroxypropylcellulose , a wax , a polyester , a polyanhydride , a polyamide , a phosphorous-based polymer , a poly(cyanoacrylate) , a polyurethane , a polyorthoester , a polydihydropyran , a polyacetal , a biodegradable poly (lipid particle, total lipid) mer , a polypeptide , a hydrogel , a carbohydrate , and combinations thereof . WO2007024323A2 CLAIM 142 . The method of claim 110 , wherein the substance to be molded is selected from the group consisting of a polymer , a solution , a monomer , a plurality of monomers , a polymerization initiator , a polymerization catalyst , an inorganic precursor , a metal precursor (lipid particle, total lipid) , a pharmaceutical agent , a tag , a magnetic material , a paramagnetic material , a ligand , a cell penetrating peptide , a porogen , a surfactant , a plurality of immiscible liquids , a solvent , and a charged species .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid (biodegradable poly, metal precursor) present in the particle .	WO2007024323A2 CLAIM 3 . The composition of claim 2 , wherein the biocompatible material is selected from the group consisting of a poly(ethylene glycol) , a poly(lactic acid) , a poly(lactic acid-co-glycolic acid) , a lactose , a phosphatidylcholine , a polylactide , a polyglycolide , a hydroxypropylcellulose , a wax , a polyester , a polyanhydride , a polyamide , a phosphorous-based polymer , a poly(cyanoacrylate) , a polyurethane , a polyorthoester , a polydihydropyran , a polyacetal , a biodegradable poly (lipid particle, total lipid) mer , a polypeptide , a hydrogel , a carbohydrate , and combinations thereof . WO2007024323A2 CLAIM 142 . The method of claim 110 , wherein the substance to be molded is selected from the group consisting of a polymer , a solution , a monomer , a plurality of monomers , a polymerization initiator , a polymerization catalyst , an inorganic precursor , a metal precursor (lipid particle, total lipid) , a pharmaceutical agent , a tag , a magnetic material , a paramagnetic material , a ligand , a cell penetrating peptide , a porogen , a surfactant , a plurality of immiscible liquids , a solvent , and a charged species .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	EP1766035A1 Filed: 2005-06-07 Issued: 2007-03-28 Lipid encapsulated interfering rna (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Protiva Biotherapeutics Inc Ian Maclachlan, Lorne R. Palmer, James Heyes
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	EP1766035A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid (cationic lipid) of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	EP1766035A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	EP1766035A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid-lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid (cationic lipid) of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 10 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation (inhibits aggregation) of particles comprises a polyethyleneglycol (PEG)-lipid conjugate .	EP1766035A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid-lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	EP1766035A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid-lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid (cationic lipid) of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation (inhibits aggregation) of particles comprises from 1 mol % to 2 mol % of the total lipid present in the particle .	EP1766035A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid-lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	EP1766035A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	EP1766035A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	EP1766035A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 19 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a median diameter (median diameter) of from about 40 nm to about 150 nm .	EP1766035A1 CLAIM 4 . The nucleic acid-lipid particle of claim 1 , wherein said particle has a median diameter (median diameter, lipid particle) of less than about 150 nm .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	EP1781593A2 Filed: 2005-06-07 Issued: 2007-05-09 Cationic lipids and methods of use (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Protiva Biotherapeutics Inc Ian Maclachlan, James Heyes, Lorne R. Palmer
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present (total lipid present) in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	EP1781593A2 CLAIM 27 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said nucleic acid (nucleic acid) is a member selected from the group consisting of : DNA , RNA , siRNA , a plasmid , an antisense oligonucleotide , a ribozyme . EP1781593A2 CLAIM 32 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	EP1781593A2 CLAIM 27 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said nucleic acid (nucleic acid) is a member selected from the group consisting of : DNA , RNA , siRNA , a plasmid , an antisense oligonucleotide , a ribozyme .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 52 mol % to 62 mol % of the total lipid present (total lipid present) in the particle .	EP1781593A2 CLAIM 32 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid present (total lipid present) in the particle .	EP1781593A2 CLAIM 32 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	EP1781593A2 CLAIM 27 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said nucleic acid (nucleic acid) is a member selected from the group consisting of : DNA , RNA , siRNA , a plasmid , an antisense oligonucleotide , a ribozyme .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	EP1781593A2 CLAIM 27 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said nucleic acid (nucleic acid) is a member selected from the group consisting of : DNA , RNA , siRNA , a plasmid , an antisense oligonucleotide , a ribozyme .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	EP1781593A2 CLAIM 27 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said nucleic acid (nucleic acid) is a member selected from the group consisting of : DNA , RNA , siRNA , a plasmid , an antisense oligonucleotide , a ribozyme .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present (total lipid present) in the particle .	EP1781593A2 CLAIM 32 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid present (total lipid present) in the particle .	EP1781593A2 CLAIM 32 . The nucleic acid-lipid particle in accordance with claim 17 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	WO2005121348A1 Filed: 2005-06-07 Issued: 2005-12-22 Lipid encapsulated interfering rna (Original Assignee) Protiva Biotherapeutics, Inc. Ian Maclachlan, Lorne R. Palmer, James Heyes
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	WO2005121348A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid (cationic lipid) of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	WO2005121348A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	WO2005121348A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid-lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid (cationic lipid) of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 10 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation (inhibits aggregation) of particles comprises a polyethyleneglycol (PEG)-lipid conjugate .	WO2005121348A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid-lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	WO2005121348A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid-lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid (cationic lipid) of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation (inhibits aggregation) of particles comprises from 1 mol % to 2 mol % of the total lipid present in the particle .	WO2005121348A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid-lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation (inhibits aggregation) of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	WO2005121348A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	WO2005121348A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	WO2005121348A1 CLAIM 1 WHAT IS CLAIMED IS : 1 . An nucleic acid-lipid particle , said nucleic acid (nucleic acid) -lipid particle comprising : (a) an interfering RNA ; (b) a cationic lipid of Formula I and having the following structure : wherein : R 1 and R 2 are independently selected from the group consisting of : H and C1-C3 alkyls ; and R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms , wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation ; (c) a non-cationic lipid ; and (d) a conjugated lipid that inhibits aggregation of particles . 2 . The nucleic acid-lipid particle of claim 1 , wherein said cationic lipid is selected from the group consisting of : l , 2-DiLinoleyloxy-N , N-dimethylaminopropane (DLinDMA) and l ,
US8058069B2 CLAIM 19 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a median diameter (median diameter) of from about 40 nm to about 150 nm .	WO2005121348A1 CLAIM 4 . The nucleic acid-lipid particle of claim 1 , wherein said particle has a median diameter (median diameter, lipid particle) of less than about 150 nm .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	CN1882693A Filed: 2004-09-15 Issued: 2006-12-20 聚乙二醇修饰的脂质化合物及其应用 (Original Assignee) 普洛体维生物治疗公司 J·海斯, I·麦克拉克伦, 埃伦格雷丝·安贝吉亚
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid ; (b) a cationic lipid (一种脂质) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	CN1882693A CLAIM 18 . 一种脂质 (cationic lipid, lipid conjugate) 体，所述脂质体包含式I的聚乙二醇-二烷氧基丙基(PEG-DAA)缀合物，式I具有如下的结构：其中：R1和R2被独立地选择并且是具有约10到约20个碳原子的烷基基团；PEG是聚乙二醇；和L是包含非酯的接头部分。
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (一种脂质) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	CN1882693A CLAIM 18 . 一种脂质 (cationic lipid, lipid conjugate) 体，所述脂质体包含式I的聚乙二醇-二烷氧基丙基(PEG-DAA)缀合物，式I具有如下的结构：其中：R1和R2被独立地选择并且是具有约10到约20个碳原子的烷基基团；PEG是聚乙二醇；和L是包含非酯的接头部分。
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (二棕榈) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	CN1882693A CLAIM 30 . 按照权利要求25的核酸-脂质颗粒，其中所述PEG-DAA缀合物是选自由PEG-二月桂基氧基丙基(C12)，PEG-二肉豆蔻基氧基丙基(C14)，PEG-二棕榈 (phospholipid comprises dipalmitoylphosphatidylcholine) 基氧基丙基(C16)，和PEG-二硬脂基氧基丙基(C18)组成的组的成员。
US8058069B2 CLAIM 13 . The nucleic acid-lipid particle of claim 12 , wherein the PEG has an average molecular weight (平均分子) of about 2 , 000 daltons .	CN1882693A CLAIM 9 . 按照权利要求1的化合物，其中所述PEG具有约550道尔顿到约10 , 000道尔顿的平均分子 (average molecular weight) 量。
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (一种脂质) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	CN1882693A CLAIM 18 . 一种脂质 (cationic lipid, lipid conjugate) 体，所述脂质体包含式I的聚乙二醇-二烷氧基丙基(PEG-DAA)缀合物，式I具有如下的结构：其中：R1和R2被独立地选择并且是具有约10到约20个碳原子的烷基基团；PEG是聚乙二醇；和L是包含非酯的接头部分。
US8058069B2 CLAIM 22 . A pharmaceutical composition (物活性) comprising a nucleic acid-lipid particle of claim 1 and a pharmaceutically acceptable carrier .	CN1882693A CLAIM 19 . 按照权利要求18的脂质体，其还包含生物活性 (pharmaceutical composition) 剂。

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	EP1664316A1 Filed: 2004-09-15 Issued: 2006-06-07 Polyethyleneglycol-modified lipid compounds and uses thereof (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Protiva Biotherapeutics Inc James Heyes, Ian Maclachlan, Ellen Grace Ambegia
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present (total lipid present) in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	EP1664316A1 CLAIM 31 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle . EP1664316A1 CLAIM 48 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said nucleic acid (nucleic acid) is DNA .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	EP1664316A1 CLAIM 48 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said nucleic acid (nucleic acid) is DNA .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 52 mol % to 62 mol % of the total lipid present (total lipid present) in the particle .	EP1664316A1 CLAIM 31 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (neutral lipid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	EP1664316A1 CLAIM 29 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said non-cationic lipid is a neutral lipid (phospholipid comprises dipalmitoylphosphatidylcholine) .
US8058069B2 CLAIM 13 . The nucleic acid-lipid particle of claim 12 , wherein the PEG has an average molecular weight (average molecular weight) of about 2 , 000 daltons .	EP1664316A1 CLAIM 9 . The compound in accordance with claim 1 , wherein said PEG has an average molecular weight (average molecular weight) ranging from about 550 daltons to about 10 , 000 daltons .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid present (total lipid present) in the particle .	EP1664316A1 CLAIM 31 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	EP1664316A1 CLAIM 48 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said nucleic acid (nucleic acid) is DNA .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	EP1664316A1 CLAIM 48 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said nucleic acid (nucleic acid) is DNA .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	EP1664316A1 CLAIM 48 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said nucleic acid (nucleic acid) is DNA .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present (total lipid present) in the particle .	EP1664316A1 CLAIM 31 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid present (total lipid present) in the particle .	EP1664316A1 CLAIM 31 . The nucleic acid-lipid particle in accordance with claim 25 , wherein said cationic lipid comprises from about 2% to about 60% of the total lipid present (total lipid present) in said particle .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	WO2005005622A2 Filed: 2004-07-09 Issued: 2005-01-20 Method of altering cell properties by administering rna (Original Assignee) Ribostem Limited Stephen Ray
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (immune function) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	WO2005005622A2 CLAIM 12 . A method according to any one of the preceding claims , wherein said method effects repair of diseased cells , alters the genetic constitution of cells , induces specific cell types and/or cell fates , changes the immunological profiles of cells , and/or induces particular desired immune function (nucleic acid) s or properties .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (immune function) comprises a small interfering RNA (siRNA) .	WO2005005622A2 CLAIM 12 . A method according to any one of the preceding claims , wherein said method effects repair of diseased cells , alters the genetic constitution of cells , induces specific cell types and/or cell fates , changes the immunological profiles of cells , and/or induces particular desired immune function (nucleic acid) s or properties .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (immune function) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	WO2005005622A2 CLAIM 12 . A method according to any one of the preceding claims , wherein said method effects repair of diseased cells , alters the genetic constitution of cells , induces specific cell types and/or cell fates , changes the immunological profiles of cells , and/or induces particular desired immune function (nucleic acid) s or properties .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (immune function) is fully encapsulated in the nucleic acid-lipid particle .	WO2005005622A2 CLAIM 12 . A method according to any one of the preceding claims , wherein said method effects repair of diseased cells , alters the genetic constitution of cells , induces specific cell types and/or cell fates , changes the immunological profiles of cells , and/or induces particular desired immune function (nucleic acid) s or properties .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (immune function) mass ratio of from about 5 to about 15 .	WO2005005622A2 CLAIM 12 . A method according to any one of the preceding claims , wherein said method effects repair of diseased cells , alters the genetic constitution of cells , induces specific cell types and/or cell fates , changes the immunological profiles of cells , and/or induces particular desired immune function (nucleic acid) s or properties .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	EP1519714A1 Filed: 2003-06-30 Issued: 2005-04-06 Method and apparatus for producing liposomes (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Protiva Biotherapeutics Inc Cory Giesbrecht, Lloyd Jeffs, Ian Maclachlan, Lorne R. Palmer
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	EP1519714A1 CLAIM 18 . The process of claim 17 , wherein said nucleic acid (nucleic acid) encodes a protein selected from the group consisting of a heφes simplex virus thymidine kinase (HSV-TK) , a cytosine deaminase , a xanthine-guaninephosphoribosyl transferase , a p53 , a purine nucleoside phosphorylase , a carboxylesterase , a deoxycytidine kinase , a nitroreductase , a thymidine phosphorylase , and a cytochrome P4502B1 .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	EP1519714A1 CLAIM 18 . The process of claim 17 , wherein said nucleic acid (nucleic acid) encodes a protein selected from the group consisting of a heφes simplex virus thymidine kinase (HSV-TK) , a cytosine deaminase , a xanthine-guaninephosphoribosyl transferase , a p53 , a purine nucleoside phosphorylase , a carboxylesterase , a deoxycytidine kinase , a nitroreductase , a thymidine phosphorylase , and a cytochrome P4502B1 .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	EP1519714A1 CLAIM 18 . The process of claim 17 , wherein said nucleic acid (nucleic acid) encodes a protein selected from the group consisting of a heφes simplex virus thymidine kinase (HSV-TK) , a cytosine deaminase , a xanthine-guaninephosphoribosyl transferase , a p53 , a purine nucleoside phosphorylase , a carboxylesterase , a deoxycytidine kinase , a nitroreductase , a thymidine phosphorylase , and a cytochrome P4502B1 .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	EP1519714A1 CLAIM 18 . The process of claim 17 , wherein said nucleic acid (nucleic acid) encodes a protein selected from the group consisting of a heφes simplex virus thymidine kinase (HSV-TK) , a cytosine deaminase , a xanthine-guaninephosphoribosyl transferase , a p53 , a purine nucleoside phosphorylase , a carboxylesterase , a deoxycytidine kinase , a nitroreductase , a thymidine phosphorylase , and a cytochrome P4502B1 .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio (salt concentration) of from about 5 to about 15 .	EP1519714A1 CLAIM 18 . The process of claim 17 , wherein said nucleic acid (nucleic acid) encodes a protein selected from the group consisting of a heφes simplex virus thymidine kinase (HSV-TK) , a cytosine deaminase , a xanthine-guaninephosphoribosyl transferase , a p53 , a purine nucleoside phosphorylase , a carboxylesterase , a deoxycytidine kinase , a nitroreductase , a thymidine phosphorylase , and a cytochrome P4502B1 . EP1519714A1 CLAIM 36 . The process of claim 28 , wherein said liposome is in a solution having a salt concentration (nucleic acid mass ratio) of about 100 mM to about 200 mM .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	CN1655764A Filed: 2003-04-04 Issued: 2005-08-17 用于给予一定大小的脂质体治疗或预防疾病的组合物和方法 (Original Assignee) 埃斯佩里安Luv发展公司 W·V·罗德里格扎, C·L·比斯盖尔
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (与一种) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	CN1655764A CLAIM 19 . 权利要求1的方法，其中所述脂质体与一种 (nucleic acid) 或多种心血管药物、抗糖尿病药物或其它治疗物质联合用药。
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (与一种) comprises a small interfering RNA (siRNA) .	CN1655764A CLAIM 19 . 权利要求1的方法，其中所述脂质体与一种 (nucleic acid) 或多种心血管药物、抗糖尿病药物或其它治疗物质联合用药。
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (二棕榈酰磷脂, 酰丝氨酸) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	CN1655764A CLAIM 12 . 权利要求1的方法，其中所述磷脂选自：卵磷脂酰胆碱，卵磷脂酰甘油，二硬脂酰磷脂酰胆碱，或二硬脂酰磷脂酰甘油，磷脂酰胆碱，磷脂酰甘油，卵磷脂，β，糖脂，γ-二棕榈酰-α-卵磷脂，鞘磷脂，磷脂酰丝氨酸 (phospholipid comprises dipalmitoylphosphatidylcholine) ，磷脂酸，N-(2，3-二(9-(Z)-十八烯基氧基))-丙-1-基-N，N，N-三甲基氯化铵，磷脂酰乙醇胺，溶血卵磷脂，溶血磷脂酰乙醇胺，磷脂酰肌醇，脑磷脂，心磷脂，脑苷脂，磷酸联十六烷基酯，二油酰磷脂酰胆碱、二棕榈酰磷脂 (phospholipid comprises dipalmitoylphosphatidylcholine) 酰胆碱，二棕榈酰磷脂酰甘油，二油酰磷脂酰甘油，棕榈酰-油酰-磷脂酰胆碱，二-硬脂酰-磷脂酰胆碱，硬脂酰-棕榈酰-磷脂酰胆碱，二-棕榈酰-磷脂酰乙醇胺，二-硬脂酰-磷脂酰乙醇胺，二-肉豆蔻酰-磷脂酰丝氨酸，二-油酰-磷脂酰胆碱，油酰-棕榈酰磷脂酰胆碱，37℃时为液晶相的脂质，以及它们的混合物。
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (与一种) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	CN1655764A CLAIM 19 . 权利要求1的方法，其中所述脂质体与一种 (nucleic acid) 或多种心血管药物、抗糖尿病药物或其它治疗物质联合用药。
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (与一种) is fully encapsulated in the nucleic acid-lipid particle .	CN1655764A CLAIM 19 . 权利要求1的方法，其中所述脂质体与一种 (nucleic acid) 或多种心血管药物、抗糖尿病药物或其它治疗物质联合用药。
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (与一种) mass ratio of from about 5 to about 15 .	CN1655764A CLAIM 19 . 权利要求1的方法，其中所述脂质体与一种 (nucleic acid) 或多种心血管药物、抗糖尿病药物或其它治疗物质联合用药。
US8058069B2 CLAIM 19 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a median diameter (平均直径) of from about 40 nm to about 150 nm .	CN1655764A CLAIM 1 . 一种预防、治疗或处理疾病或机体病症的方法，所述方法包括以单剂量或分剂量给予50mg/kg-300mg/kg的脂质体，所述脂质体包含磷脂和水层，所述脂质体的平均直径 (median diameter) 为约50nm-250nm。

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	WO9914346A2 Filed: 1998-09-18 Issued: 1999-03-25 SENSE mRNA THERAPY (Original Assignee) Sequitur, Inc. Tod M. Woolf, Kristin Wiederholt, Margaret Taylor
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	WO9914346A2 CLAIM 14 . The mRNA molecule of claim 11 , wherein the agent is a nucleic acid (nucleic acid) molecule . WO9914346A2 CLAIM 27 . The mRNA molecule of claim 1 , wherein said delivery vehicle is selected from the group consisting of : cationic lipid (cationic lipid) containing complexes , uncharged lipids , nanoparticles .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) comprises a small interfering RNA (siRNA) .	WO9914346A2 CLAIM 14 . The mRNA molecule of claim 11 , wherein the agent is a nucleic acid (nucleic acid) molecule .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	WO9914346A2 CLAIM 27 . The mRNA molecule of claim 1 , wherein said delivery vehicle is selected from the group consisting of : cationic lipid (cationic lipid) containing complexes , uncharged lipids , nanoparticles .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	WO9914346A2 CLAIM 27 . The mRNA molecule of claim 1 , wherein said delivery vehicle is selected from the group consisting of : cationic lipid (cationic lipid) containing complexes , uncharged lipids , nanoparticles .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	WO9914346A2 CLAIM 14 . The mRNA molecule of claim 11 , wherein the agent is a nucleic acid (nucleic acid) molecule .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	WO9914346A2 CLAIM 14 . The mRNA molecule of claim 11 , wherein the agent is a nucleic acid (nucleic acid) molecule .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (nucleic acid) mass ratio of from about 5 to about 15 .	WO9914346A2 CLAIM 14 . The mRNA molecule of claim 11 , wherein the agent is a nucleic acid (nucleic acid) molecule .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	WO9914226A2 Filed: 1998-09-14 Issued: 1999-03-25 Bi- and tri-cyclic nucleoside, nucleotide and oligonucleotide analogues (Original Assignee) Exiqon A/S Jesper Wengel, Poul Nielsen
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (nucleic acid amplification reaction, stranded nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	WO9914226A2 CLAIM 1 19 . The use according to claim 1 15 for capture and detection of naturally occurring or synthetic double stranded or single stranded nucleic acid (nucleic acid, nucleic acid mass ratio) s such as RNA or DNA . WO9914226A2 CLAIM 1 24 . The use according to claim 1 15 as a primer in a nucleic acid amplification reaction (nucleic acid, nucleic acid mass ratio) .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid amplification reaction, stranded nucleic acid) comprises a small interfering RNA (siRNA) .	WO9914226A2 CLAIM 1 19 . The use according to claim 1 15 for capture and detection of naturally occurring or synthetic double stranded or single stranded nucleic acid (nucleic acid, nucleic acid mass ratio) s such as RNA or DNA . WO9914226A2 CLAIM 1 24 . The use according to claim 1 15 as a primer in a nucleic acid amplification reaction (nucleic acid, nucleic acid mass ratio) .
US8058069B2 CLAIM 13 . The nucleic acid-lipid particle of claim 12 , wherein the PEG has an average molecular weight (following criteria) of about 2 , 000 daltons .	WO9914226A2 CLAIM 13 . An oligomer according to claim 1 2 , wherein one of the following criteria (average molecular weight) applies for at least one LNA ; (i) R 2* and R 4* together designate a biradical selected from -0- , -S- , -N(R )- , - (CR " ; R ) r+ε+ r , -(CR * R ' ; ) r -0-(CR * R ) ε - , -(CR ' ; R ) r -S-(CR ' ; R ' ; ) ε - , -(CR * R ) r -N(R )- (CR * R ) 6 - , -0-(CR R ) r+ε -0- , -S-(CR R ) r+ε -0- , -0-(CR R ) r+ε -S- , -N(R (CR R ' ; ) r+ε -0- , -0-(CR ' ; R ) r+6 -N(R )- , -S-(CR * R ) r+β -S- , -N(R )-(CR * R ) r+ε -N(R )- , -N(R )-(CR R ) r+ε -S- , and -S-(CR R ) r+ε -N(R )- ; (ii) R 2 and R 3 together designate a biradical selected from -0- , -(CR * R ) r+s - , -(CR R " ; ) r -0-(CR ' ; R ) β - , -(CR R ) r -S-(CR R ) β - , and -(CR R ) r -N(R )-(CR * R ) ε - ; (iii) R 2 and R 3 together designate a biradical selected from -0- , -(CR * R ) r+ε - , -(CR R ) r -0-(CR R ) ε - , -(CR R ) r -S-(CR R ) β - , and -(CR R " ; ) r -N(R )-(CR R ) ε - ; (iv) R 3 and R 4 together designate a biradical selected from -(CR * R ) r -0-(CR R ) ε - , -(CR R ) r -S-(CR R ) 6 - , and -(CR R ) r -N(R )-(CR * R ) ε - ; 5 (v) R 3 and R 5 together designate a biradical selected from -(CR R ) r -0-(CR R ) ε - , -(CR R ) r -S-(CR R ) 6 - , and -(CR R ) r -N(R )-(CR * R ) ε - ; or (vi) R 1 and R 4* together designate a biradical selected from -(CR * R ) r -0- (CR R ) ε - , -(CR R ) r -S-(CR R ) ε - , and -(CR R ) r -N(R )-(CR * R ) ε - ; (vii) R 1 and R 2* together designate a biradical selected from -(CR * R ) r -0- 0 (CR R ) ε - , -(CR R ) r -S-(CR R ) ε - , and -(CR R ) r -N(R )-(CR * R *) ε - ; wherein each of r and s is 0-3 with the proviso that the sum r + s is 1 -4 , and where X is selected from -0- , -S- , and -N(R H)- where R H designates hydrogen or C^-alkyl .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid amplification reaction, stranded nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	WO9914226A2 CLAIM 1 19 . The use according to claim 1 15 for capture and detection of naturally occurring or synthetic double stranded or single stranded nucleic acid (nucleic acid, nucleic acid mass ratio) s such as RNA or DNA . WO9914226A2 CLAIM 1 24 . The use according to claim 1 15 as a primer in a nucleic acid amplification reaction (nucleic acid, nucleic acid mass ratio) .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (nucleic acid amplification reaction, stranded nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	WO9914226A2 CLAIM 1 19 . The use according to claim 1 15 for capture and detection of naturally occurring or synthetic double stranded or single stranded nucleic acid (nucleic acid, nucleic acid mass ratio) s such as RNA or DNA . WO9914226A2 CLAIM 1 24 . The use according to claim 1 15 as a primer in a nucleic acid amplification reaction (nucleic acid, nucleic acid mass ratio) .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (nucleic acid amplification reaction, stranded nucleic acid) mass ratio of from about 5 to about 15 .	WO9914226A2 CLAIM 1 19 . The use according to claim 1 15 for capture and detection of naturally occurring or synthetic double stranded or single stranded nucleic acid (nucleic acid, nucleic acid mass ratio) s such as RNA or DNA . WO9914226A2 CLAIM 1 24 . The use according to claim 1 15 as a primer in a nucleic acid amplification reaction (nucleic acid, nucleic acid mass ratio) .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US6376248B1 Filed: 1998-03-16 Issued: 2002-04-23 Peptide-enhanced transfections (Original Assignee) Life Technologies Inc (Current Assignee) Life Technologies Corp Pamela Hawley-Nelson, Jianqing Lan, PoJen Shih, Joel A. Jessee, Kevin P. Schifferli, Gulilat Gebeyehu, Valentina C. Ciccarone, Krista L. Evans
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US6376248B1 CLAIM 16 . The composition of claim 15 , wherein said nucleic acid (nucleic acid) binding groups comprise at least one polyamine .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	US6376248B1 CLAIM 16 . The composition of claim 15 , wherein said nucleic acid (nucleic acid) binding groups comprise at least one polyamine .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (neutral lipid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	US6376248B1 CLAIM 6 . The composition of claim 5 , wherein said transfection agents further comprise one or more neutral lipid (phospholipid comprises dipalmitoylphosphatidylcholine) s .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US6376248B1 CLAIM 16 . The composition of claim 15 , wherein said nucleic acid (nucleic acid) binding groups comprise at least one polyamine .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	US6376248B1 CLAIM 16 . The composition of claim 15 , wherein said nucleic acid (nucleic acid) binding groups comprise at least one polyamine .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	US6376248B1 CLAIM 16 . The composition of claim 15 , wherein said nucleic acid (nucleic acid) binding groups comprise at least one polyamine .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US6004573A Filed: 1997-10-03 Issued: 1999-12-21 Biodegradable low molecular weight triblock poly(lactide-co-glycolide) polyethylene glycol copolymers having reverse thermal gelation properties (Original Assignee) MacroMed Inc (Current Assignee) JG CONSULTING AG ; KIM PHD SUNG WAN ; BTG International Ltd Ramesh C. Rathi, Gaylen M. Zentner
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present (mole percent) in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US6004573A CLAIM 3 . An aqueous polymeric composition according to claim 2 wherein the PLGA copolymer A-block comprises between about 65 to 85 mole percent (total lipid present) lactide and between about 15 and 35 mole percent glycolide .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 52 mol % to 62 mol % of the total lipid present (mole percent) in the particle .	US6004573A CLAIM 3 . An aqueous polymeric composition according to claim 2 wherein the PLGA copolymer A-block comprises between about 65 to 85 mole percent (total lipid present) lactide and between about 15 and 35 mole percent glycolide .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (polymeric drug) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	US6004573A CLAIM 1 . An aqueous biodegradable polymeric drug (phospholipid comprises dipalmitoylphosphatidylcholine) delivery composition possessing reverse thermal gelation properties comprised of an aqueous phase having uniformly contained therein : (a) an effective amount of a drug ; and (b) a biodegradable ABA-type block copolymer of the formula : PLGA-PEG-PLGA where PLGA is a hydrophobic poly(lactide-co-glycolide) copolymer that comprises the A-blocks and PEG is a hydrophilic polyethylene glycol polymer that comprises the B-block , said block copolymer having an average molecular weight of between about 3100 and 4500 wherein , in the block copolymer , the PLGA A-blocks comprise about 51 to 83% by weight of said copolymer and the PEG B-block comprises about 17 to 49% by weight of said copolymer .
US8058069B2 CLAIM 13 . The nucleic acid-lipid particle of claim 12 , wherein the PEG has an average molecular weight (average molecular weight) of about 2 , 000 daltons .	US6004573A CLAIM 1 . An aqueous biodegradable polymeric drug delivery composition possessing reverse thermal gelation properties comprised of an aqueous phase having uniformly contained therein : (a) an effective amount of a drug ; and (b) a biodegradable ABA-type block copolymer of the formula : PLGA-PEG-PLGA where PLGA is a hydrophobic poly(lactide-co-glycolide) copolymer that comprises the A-blocks and PEG is a hydrophilic polyethylene glycol polymer that comprises the B-block , said block copolymer having an average molecular weight (average molecular weight) of between about 3100 and 4500 wherein , in the block copolymer , the PLGA A-blocks comprise about 51 to 83% by weight of said copolymer and the PEG B-block comprises about 17 to 49% by weight of said copolymer .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid present (mole percent) in the particle .	US6004573A CLAIM 3 . An aqueous polymeric composition according to claim 2 wherein the PLGA copolymer A-block comprises between about 65 to 85 mole percent (total lipid present) lactide and between about 15 and 35 mole percent glycolide .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present (mole percent) in the particle .	US6004573A CLAIM 3 . An aqueous polymeric composition according to claim 2 wherein the PLGA copolymer A-block comprises between about 65 to 85 mole percent (total lipid present) lactide and between about 15 and 35 mole percent glycolide .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid present (mole percent) in the particle .	US6004573A CLAIM 3 . An aqueous polymeric composition according to claim 2 wherein the PLGA copolymer A-block comprises between about 65 to 85 mole percent (total lipid present) lactide and between about 15 and 35 mole percent glycolide .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US5789554A Filed: 1996-07-31 Issued: 1998-08-04 Immunoconjugates and humanized antibodies specific for B-cell lymphoma and leukemia cells (Original Assignee) Immunomedics Inc (Current Assignee) Immunomedics Inc Shui-on Leung, Hans Hansen
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present (diagnostic reagent) in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation (selecting cells) of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US5789554A CLAIM 3 . A conjugate according to claim 2 wherein said diagnostic reagent (lipid particle, total lipid present) comprises a label . US5789554A CLAIM 31 . A method for the expression of a humanized LL2 monoclonal antibody , designated hLL2-2 , comprising : (a) linearizing the expression vector of claim 17 and an expression vector comprising a DNA sequence encoding the humanized hLL2 light chain variable region of SEQ ID NO : 6 ; (b) transfecting mammalian cells with said linearized vectors ; (c) selecting said transfected cells which express a xanthine-guanine phosphoribosyltransferase (gpt) gene ; and (d) selecting cells (inhibits aggregation) secreting said humanized monoclonal antibody from said cells which express said gpt gene .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 52 mol % to 62 mol % of the total lipid present (diagnostic reagent) in the particle .	US5789554A CLAIM 3 . A conjugate according to claim 2 wherein said diagnostic reagent (lipid particle, total lipid present) comprises a label .
US8058069B2 CLAIM 10 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation (selecting cells) of particles comprises a polyethyleneglycol (PEG)-lipid conjugate .	US5789554A CLAIM 31 . A method for the expression of a humanized LL2 monoclonal antibody , designated hLL2-2 , comprising : (a) linearizing the expression vector of claim 17 and an expression vector comprising a DNA sequence encoding the humanized hLL2 light chain variable region of SEQ ID NO : 6 ; (b) transfecting mammalian cells with said linearized vectors ; (c) selecting said transfected cells which express a xanthine-guanine phosphoribosyltransferase (gpt) gene ; and (d) selecting cells (inhibits aggregation) secreting said humanized monoclonal antibody from said cells which express said gpt gene .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation (selecting cells) of particles comprises from 1 mol % to 2 mol % of the total lipid present (diagnostic reagent) in the particle .	US5789554A CLAIM 3 . A conjugate according to claim 2 wherein said diagnostic reagent (lipid particle, total lipid present) comprises a label . US5789554A CLAIM 31 . A method for the expression of a humanized LL2 monoclonal antibody , designated hLL2-2 , comprising : (a) linearizing the expression vector of claim 17 and an expression vector comprising a DNA sequence encoding the humanized hLL2 light chain variable region of SEQ ID NO : 6 ; (b) transfecting mammalian cells with said linearized vectors ; (c) selecting said transfected cells which express a xanthine-guanine phosphoribosyltransferase (gpt) gene ; and (d) selecting cells (inhibits aggregation) secreting said humanized monoclonal antibody from said cells which express said gpt gene .
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid present (diagnostic reagent) in the particle .	US5789554A CLAIM 3 . A conjugate according to claim 2 wherein said diagnostic reagent (lipid particle, total lipid present) comprises a label .
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid present (diagnostic reagent) in the particle .	US5789554A CLAIM 3 . A conjugate according to claim 2 wherein said diagnostic reagent (lipid particle, total lipid present) comprises a label .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	WO9640964A2 Filed: 1996-06-06 Issued: 1996-12-19 Lipid-nucleic acid particles prepared via a hydrophobic lipid-nucleic acid complex intermediate and use for gene transfer (Original Assignee) Inex Pharmaceuticals Corporation Jeffery J. Wheeler, Marcel B. Bally, Yuan-Peng Zhang, Dorothy L. Reimer, Michael Hope, Pieter R. Cullis, Peter Scherrer
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	WO9640964A2 CLAIM 2 . A complex in accordance with claim 1 , wherein said nucleic acid (nucleic acid) is a plasmid .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	WO9640964A2 CLAIM 2 . A complex in accordance with claim 1 , wherein said nucleic acid (nucleic acid) is a plasmid .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	WO9640964A2 CLAIM 2 . A complex in accordance with claim 1 , wherein said nucleic acid (nucleic acid) is a plasmid .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	WO9640964A2 CLAIM 2 . A complex in accordance with claim 1 , wherein said nucleic acid (nucleic acid) is a plasmid .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	WO9640964A2 CLAIM 2 . A complex in accordance with claim 1 , wherein said nucleic acid (nucleic acid) is a plasmid .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	US5716785A Filed: 1996-04-19 Issued: 1998-02-10 Processes for genetic manipulations using promoters (Original Assignee) Leland Stanford Junior University (Current Assignee) Leland Stanford Junior University Russell N. Van Gelder, Mark E. Von Zastrow, Jack D. Barchas, James H. Eberwine
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (stranded nucleic acid, molar excess) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	US5716785A CLAIM 3 . A process for amplifying at least one target nucleic acid sequence comprising : (a) synthesizing a double-stranded nucleic acid (nucleic acid, nucleic acid mass ratio) by : i) hybridizing a primer complex to the target nucleic acid sequence and extending the primer complex to form a first DNA strand complementary to the target sequence , wherein said primer complex comprises a promoter and a primer region complementary to the target nucleic acid sequence , and ii) synthesizing a second DNA strand complementary to the first DNA strand without using an exogenous primer complementary to the first DNA strand ; and , (b) transcribing copies of RNA initiated from the promoter region of the primer complex , wherein said copies of RNA are complementary to the second DNA strand . US5716785A CLAIM 16 . A method for subtractive hybridization comprising the steps : (a) binding a primer to sense RNA molecules in a first population , wherein the primer is operably linked to a promoter sequence in an anti-sense orientation ; (b) synthesizing a first complementary DNA (cDNA) strand by elongation from the primer ; (c) synthesizing a second cDNA strand without using an exogenous primer complementary to the first strand , whereupon a functional promoter is generated ; (d) initiating RNA synthesis from the promoter by adding RNA polymerase , whereby anti-sense RNA (aRNA) is produced ; (e) introducing the aRNA in molar excess (nucleic acid, nucleic acid mass ratio) to a second population of sense RNA molecules , whereby complementary RNA sequences from the two populations hybridize ; and (f) isolating remaining single-stranded sense RNA .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (stranded nucleic acid, molar excess) comprises a small interfering RNA (siRNA) .	US5716785A CLAIM 3 . A process for amplifying at least one target nucleic acid sequence comprising : (a) synthesizing a double-stranded nucleic acid (nucleic acid, nucleic acid mass ratio) by : i) hybridizing a primer complex to the target nucleic acid sequence and extending the primer complex to form a first DNA strand complementary to the target sequence , wherein said primer complex comprises a promoter and a primer region complementary to the target nucleic acid sequence , and ii) synthesizing a second DNA strand complementary to the first DNA strand without using an exogenous primer complementary to the first DNA strand ; and , (b) transcribing copies of RNA initiated from the promoter region of the primer complex , wherein said copies of RNA are complementary to the second DNA strand . US5716785A CLAIM 16 . A method for subtractive hybridization comprising the steps : (a) binding a primer to sense RNA molecules in a first population , wherein the primer is operably linked to a promoter sequence in an anti-sense orientation ; (b) synthesizing a first complementary DNA (cDNA) strand by elongation from the primer ; (c) synthesizing a second cDNA strand without using an exogenous primer complementary to the first strand , whereupon a functional promoter is generated ; (d) initiating RNA synthesis from the promoter by adding RNA polymerase , whereby anti-sense RNA (aRNA) is produced ; (e) introducing the aRNA in molar excess (nucleic acid, nucleic acid mass ratio) to a second population of sense RNA molecules , whereby complementary RNA sequences from the two populations hybridize ; and (f) isolating remaining single-stranded sense RNA .
US8058069B2 CLAIM 13 . The nucleic acid-lipid particle of claim 12 , wherein the PEG has an average molecular weight (human tissue) of about 2 , 000 daltons .	US5716785A CLAIM 20 . A method according to claim 18 , wherein the cells are obtained from human tissue (average molecular weight) .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (stranded nucleic acid, molar excess) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	US5716785A CLAIM 3 . A process for amplifying at least one target nucleic acid sequence comprising : (a) synthesizing a double-stranded nucleic acid (nucleic acid, nucleic acid mass ratio) by : i) hybridizing a primer complex to the target nucleic acid sequence and extending the primer complex to form a first DNA strand complementary to the target sequence , wherein said primer complex comprises a promoter and a primer region complementary to the target nucleic acid sequence , and ii) synthesizing a second DNA strand complementary to the first DNA strand without using an exogenous primer complementary to the first DNA strand ; and , (b) transcribing copies of RNA initiated from the promoter region of the primer complex , wherein said copies of RNA are complementary to the second DNA strand . US5716785A CLAIM 16 . A method for subtractive hybridization comprising the steps : (a) binding a primer to sense RNA molecules in a first population , wherein the primer is operably linked to a promoter sequence in an anti-sense orientation ; (b) synthesizing a first complementary DNA (cDNA) strand by elongation from the primer ; (c) synthesizing a second cDNA strand without using an exogenous primer complementary to the first strand , whereupon a functional promoter is generated ; (d) initiating RNA synthesis from the promoter by adding RNA polymerase , whereby anti-sense RNA (aRNA) is produced ; (e) introducing the aRNA in molar excess (nucleic acid, nucleic acid mass ratio) to a second population of sense RNA molecules , whereby complementary RNA sequences from the two populations hybridize ; and (f) isolating remaining single-stranded sense RNA .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (stranded nucleic acid, molar excess) is fully encapsulated in the nucleic acid-lipid particle .	US5716785A CLAIM 3 . A process for amplifying at least one target nucleic acid sequence comprising : (a) synthesizing a double-stranded nucleic acid (nucleic acid, nucleic acid mass ratio) by : i) hybridizing a primer complex to the target nucleic acid sequence and extending the primer complex to form a first DNA strand complementary to the target sequence , wherein said primer complex comprises a promoter and a primer region complementary to the target nucleic acid sequence , and ii) synthesizing a second DNA strand complementary to the first DNA strand without using an exogenous primer complementary to the first DNA strand ; and , (b) transcribing copies of RNA initiated from the promoter region of the primer complex , wherein said copies of RNA are complementary to the second DNA strand . US5716785A CLAIM 16 . A method for subtractive hybridization comprising the steps : (a) binding a primer to sense RNA molecules in a first population , wherein the primer is operably linked to a promoter sequence in an anti-sense orientation ; (b) synthesizing a first complementary DNA (cDNA) strand by elongation from the primer ; (c) synthesizing a second cDNA strand without using an exogenous primer complementary to the first strand , whereupon a functional promoter is generated ; (d) initiating RNA synthesis from the promoter by adding RNA polymerase , whereby anti-sense RNA (aRNA) is produced ; (e) introducing the aRNA in molar excess (nucleic acid, nucleic acid mass ratio) to a second population of sense RNA molecules , whereby complementary RNA sequences from the two populations hybridize ; and (f) isolating remaining single-stranded sense RNA .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (stranded nucleic acid, molar excess) mass ratio of from about 5 to about 15 .	US5716785A CLAIM 3 . A process for amplifying at least one target nucleic acid sequence comprising : (a) synthesizing a double-stranded nucleic acid (nucleic acid, nucleic acid mass ratio) by : i) hybridizing a primer complex to the target nucleic acid sequence and extending the primer complex to form a first DNA strand complementary to the target sequence , wherein said primer complex comprises a promoter and a primer region complementary to the target nucleic acid sequence , and ii) synthesizing a second DNA strand complementary to the first DNA strand without using an exogenous primer complementary to the first DNA strand ; and , (b) transcribing copies of RNA initiated from the promoter region of the primer complex , wherein said copies of RNA are complementary to the second DNA strand . US5716785A CLAIM 16 . A method for subtractive hybridization comprising the steps : (a) binding a primer to sense RNA molecules in a first population , wherein the primer is operably linked to a promoter sequence in an anti-sense orientation ; (b) synthesizing a first complementary DNA (cDNA) strand by elongation from the primer ; (c) synthesizing a second cDNA strand without using an exogenous primer complementary to the first strand , whereupon a functional promoter is generated ; (d) initiating RNA synthesis from the promoter by adding RNA polymerase , whereby anti-sense RNA (aRNA) is produced ; (e) introducing the aRNA in molar excess (nucleic acid, nucleic acid mass ratio) to a second population of sense RNA molecules , whereby complementary RNA sequences from the two populations hybridize ; and (f) isolating remaining single-stranded sense RNA .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	JPH0867666A Filed: 1994-08-29 Issued: 1996-03-12 カロチノイド含有粉末製剤及びその製造方法 (Original Assignee) Lion Corp; ライオン株式会社 Masahiko Furue, Hiroshige Hamakawa, Tomoko Sato, 智子佐藤, 雅彦古江, 弘茂浜川
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid (の合計) present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	JPH0867666A CLAIM 5 【請求項５】カロチノイドがパーム油を抽出して得られ、β−カロチンを５５〜７０重量％、α−カロチンを２０〜４０重量％、β−カロチンとα−カロチンの合計 (total lipid) 量として８５〜９７重量％を含有するカロチノイドである請求項４記載の製造方法。
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid comprises a small interfering (カロチン) RNA (siRNA) .	JPH0867666A CLAIM 2 【請求項２】カロチノイドが、パーム油を抽出して得られ、β−カロチン (small interfering) を５５〜７０重量％、α−カロチンを２０〜４０重量％、β−カロチンとα−カロチンの合計量として８５〜９７重量％を含有するカロチノイドである請求項１記載の製剤。
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid comprises from 52 mol % to 62 mol % of the total lipid (の合計) present in the particle .	JPH0867666A CLAIM 5 【請求項５】カロチノイドがパーム油を抽出して得られ、β−カロチンを５５〜７０重量％、α−カロチンを２０〜４０重量％、β−カロチンとα−カロチンの合計 (total lipid) 量として８５〜９７重量％を含有するカロチノイドである請求項４記載の製造方法。
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles comprises from 1 mol % to 2 mol % of the total lipid (の合計) present in the particle .	JPH0867666A CLAIM 5 【請求項５】カロチノイドがパーム油を抽出して得られ、β−カロチンを５５〜７０重量％、α−カロチンを２０〜４０重量％、β−カロチンとα−カロチンの合計 (total lipid) 量として８５〜９７重量％を含有するカロチノイドである請求項４記載の製造方法。
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid mass ratio (重量比) of from about 5 to about 15 .	JPH0867666A CLAIM 1 【請求項１】カロチノイドと、油脂と、水溶性抗酸化剤及び油溶性抗酸化剤とを含有し、水溶性抗酸化剤が０．５〜８重量％であって、水溶性抗酸化剤と油溶性抗酸化剤との割合が重量比 (nucleic acid mass ratio) で１：１〜１：１０であり、かつ平均粒子径が１０〜２００μｍの粉末からなることを特徴とするカロチノイド含有粉末製剤。
US8058069B2 CLAIM 20 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises from 5 mol % to 9 mol % of the total lipid (の合計) present in the particle .	JPH0867666A CLAIM 5 【請求項５】カロチノイドがパーム油を抽出して得られ、β−カロチンを５５〜７０重量％、α−カロチンを２０〜４０重量％、β−カロチンとα−カロチンの合計 (total lipid) 量として８５〜９７重量％を含有するカロチノイドである請求項４記載の製造方法。
US8058069B2 CLAIM 21 . The nucleic acid-lipid particle of claim 1 , wherein the cholesterol or derivative thereof comprises from 32 mol % to 36 mol % of the total lipid (の合計) present in the particle .	JPH0867666A CLAIM 5 【請求項５】カロチノイドがパーム油を抽出して得られ、β−カロチンを５５〜７０重量％、α−カロチンを２０〜４０重量％、β−カロチンとα−カロチンの合計 (total lipid) 量として８５〜９７重量％を含有するカロチノイドである請求項４記載の製造方法。
US8058069B2 CLAIM 22 . A pharmaceutical composition (型乳化物) comprising a nucleic acid-lipid particle of claim 1 and a pharmaceutically acceptable carrier .	JPH0867666A CLAIM 4 【請求項４】カロチノイドを油脂に混合すると共に、油溶性抗酸化剤を添加した油相を、水溶性抗酸化剤をこの水溶性抗酸化剤と上記油溶性抗酸化剤との重量比が１：１〜１：１０となるように含む水相に乳化して平均粒子径が１００〜５００ｎｍのＯ／Ｗ型乳化物 (pharmaceutical composition) を調製し、これを乾燥粉末化して平均粒子径が１０〜２００μ ｍの粉末を得ることを特徴とするカロチノイド含有粉末製剤の製造方法。

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	WO9502698A1 Filed: 1994-07-12 Issued: 1995-01-26 Composition and methods for transfecting eukaryotic cells (Original Assignee) Life Technologies, Inc. Joel A. Jessee
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	WO9502698A1 CLAIM 1 . A composition for transfecting a eukaryotic cell which comprises a nucleic acid , a cationic lipid capable of complexing said nucleic acid (nucleic acid) , and a viral agent which is an active or inactive enveloped virus or a component of an enveloped virus .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	WO9502698A1 CLAIM 1 . A composition for transfecting a eukaryotic cell which comprises a nucleic acid , a cationic lipid capable of complexing said nucleic acid (nucleic acid) , and a viral agent which is an active or inactive enveloped virus or a component of an enveloped virus .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (neutral lipid) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	WO9502698A1 CLAIM 4 . The composition of claim 1 further comprising a neutral lipid (phospholipid comprises dipalmitoylphosphatidylcholine) .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	WO9502698A1 CLAIM 1 . A composition for transfecting a eukaryotic cell which comprises a nucleic acid , a cationic lipid capable of complexing said nucleic acid (nucleic acid) , and a viral agent which is an active or inactive enveloped virus or a component of an enveloped virus .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	WO9502698A1 CLAIM 1 . A composition for transfecting a eukaryotic cell which comprises a nucleic acid , a cationic lipid capable of complexing said nucleic acid (nucleic acid) , and a viral agent which is an active or inactive enveloped virus or a component of an enveloped virus .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	WO9502698A1 CLAIM 1 . A composition for transfecting a eukaryotic cell which comprises a nucleic acid , a cationic lipid capable of complexing said nucleic acid (nucleic acid) , and a viral agent which is an active or inactive enveloped virus or a component of an enveloped virus .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	JPH07242568A Filed: 1994-03-04 Issued: 1995-09-19 苦味隠蔽製剤 (Original Assignee) Eisai Co Ltd; エーザイ株式会社 Masao Kawamura, Katsuhiro Nakamura, Shigemitsu Osawa, Satoshi Sugawara, 雄啓中村, 重光大沢, 政男河村, 智菅原
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (溶解性) ; (b) a cationic lipid (硫酸ナトリウム) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	JPH07242568A CLAIM 3 【請求項３】界面活性剤が、ソルビタン脂肪酸エステル、ショ糖脂肪酸エステル、ポリオキシエチレンソルビタン脂肪酸エステル、ポリエチレングリコール、ポリオキシエチレン硬化ヒマシ油、ポリオキシプロピレングリコール、ポリオキシエチレン−ポリオキシプロピレングリコール、ラウリル硫酸ナトリウム (cationic lipid) からなる群より選ばれる１種または２種以上の界面活性剤である請求項１記載の組成物。 JPH07242568A CLAIM 4 【請求項４】導水剤が、水溶解性 (nucleic acid) 、水分散性若しくは水膨潤性物質である請求項１記載の組成物。
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (溶解性) comprises a small interfering RNA (siRNA) .	JPH07242568A CLAIM 4 【請求項４】導水剤が、水溶解性 (nucleic acid) 、水分散性若しくは水膨潤性物質である請求項１記載の組成物。
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (硫酸ナトリウム) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	JPH07242568A CLAIM 3 【請求項３】界面活性剤が、ソルビタン脂肪酸エステル、ショ糖脂肪酸エステル、ポリオキシエチレンソルビタン脂肪酸エステル、ポリエチレングリコール、ポリオキシエチレン硬化ヒマシ油、ポリオキシプロピレングリコール、ポリオキシエチレン−ポリオキシプロピレングリコール、ラウリル硫酸ナトリウム (cationic lipid) からなる群より選ばれる１種または２種以上の界面活性剤である請求項１記載の組成物。
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (硫酸ナトリウム) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	JPH07242568A CLAIM 3 【請求項３】界面活性剤が、ソルビタン脂肪酸エステル、ショ糖脂肪酸エステル、ポリオキシエチレンソルビタン脂肪酸エステル、ポリエチレングリコール、ポリオキシエチレン硬化ヒマシ油、ポリオキシプロピレングリコール、ポリオキシエチレン−ポリオキシプロピレングリコール、ラウリル硫酸ナトリウム (cationic lipid) からなる群より選ばれる１種または２種以上の界面活性剤である請求項１記載の組成物。
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (溶解性) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	JPH07242568A CLAIM 4 【請求項４】導水剤が、水溶解性 (nucleic acid) 、水分散性若しくは水膨潤性物質である請求項１記載の組成物。
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (溶解性) is fully encapsulated in the nucleic acid-lipid particle .	JPH07242568A CLAIM 4 【請求項４】導水剤が、水溶解性 (nucleic acid) 、水分散性若しくは水膨潤性物質である請求項１記載の組成物。
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (溶解性) mass ratio of from about 5 to about 15 .	JPH07242568A CLAIM 4 【請求項４】導水剤が、水溶解性 (nucleic acid) 、水分散性若しくは水膨潤性物質である請求項１記載の組成物。

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	WO9324640A2 Filed: 1993-06-04 Issued: 1993-12-09 Methods and compositions for in vivo gene therapy (Original Assignee) The Regents Of The University Of California Robert J. Debs, Ning Zhu
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid (said nucleic acid) ; (b) a cationic lipid (cationic lipid) comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	WO9324640A2 CLAIM 1 . A complex of an expression cassette comprising : as operably joined components , a transcriptional and translational initiation regulatory region , a nucleic acid sequence of interest , and a transcriptional termination regulatory region , wherein said regulatory regions are functional in the cells of a mammalian host , and optionally , a cationic lipid (cationic lipid) carrier , and wherein said nucleic acid (nucleic acid) sequence of interest is free of introns .
US8058069B2 CLAIM 2 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) comprises a small interfering RNA (siRNA) .	WO9324640A2 CLAIM 1 . A complex of an expression cassette comprising : as operably joined components , a transcriptional and translational initiation regulatory region , a nucleic acid sequence of interest , and a transcriptional termination regulatory region , wherein said regulatory regions are functional in the cells of a mammalian host , and optionally , a cationic lipid carrier , and wherein said nucleic acid (nucleic acid) sequence of interest is free of introns .
US8058069B2 CLAIM 8 . The nucleic acid-lipid particle of claim 1 , wherein the cationic lipid (cationic lipid) comprises from 52 mol % to 62 mol % of the total lipid present in the particle .	WO9324640A2 CLAIM 1 . A complex of an expression cassette comprising : as operably joined components , a transcriptional and translational initiation regulatory region , a nucleic acid sequence of interest , and a transcriptional termination regulatory region , wherein said regulatory regions are functional in the cells of a mammalian host , and optionally , a cationic lipid (cationic lipid) carrier , and wherein said nucleic acid sequence of interest is free of introns .
US8058069B2 CLAIM 14 . The nucleic acid-lipid particle of claim 10 , wherein the nucleic acid-lipid particle comprises about 57 . 1 mol % cationic lipid (cationic lipid) , about 7 . 1 mol % phospholipid , about 34 . 3 mol % cholesterol or a derivative thereof , and about 1 . 4 mol % PEG-lipid conjugate .	WO9324640A2 CLAIM 1 . A complex of an expression cassette comprising : as operably joined components , a transcriptional and translational initiation regulatory region , a nucleic acid sequence of interest , and a transcriptional termination regulatory region , wherein said regulatory regions are functional in the cells of a mammalian host , and optionally , a cationic lipid (cationic lipid) carrier , and wherein said nucleic acid sequence of interest is free of introns .
US8058069B2 CLAIM 16 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) in the nucleic acid-lipid particle is not substantially degraded after incubation of the particle in serum at 37° C . for 30 minutes .	WO9324640A2 CLAIM 1 . A complex of an expression cassette comprising : as operably joined components , a transcriptional and translational initiation regulatory region , a nucleic acid sequence of interest , and a transcriptional termination regulatory region , wherein said regulatory regions are functional in the cells of a mammalian host , and optionally , a cationic lipid carrier , and wherein said nucleic acid (nucleic acid) sequence of interest is free of introns .
US8058069B2 CLAIM 17 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid (said nucleic acid) is fully encapsulated in the nucleic acid-lipid particle .	WO9324640A2 CLAIM 1 . A complex of an expression cassette comprising : as operably joined components , a transcriptional and translational initiation regulatory region , a nucleic acid sequence of interest , and a transcriptional termination regulatory region , wherein said regulatory regions are functional in the cells of a mammalian host , and optionally , a cationic lipid carrier , and wherein said nucleic acid (nucleic acid) sequence of interest is free of introns .
US8058069B2 CLAIM 18 . The nucleic acid-lipid particle of claim 1 , wherein the nucleic acid-lipid particle has a lipid : nucleic acid (said nucleic acid) mass ratio of from about 5 to about 15 .	WO9324640A2 CLAIM 1 . A complex of an expression cassette comprising : as operably joined components , a transcriptional and translational initiation regulatory region , a nucleic acid sequence of interest , and a transcriptional termination regulatory region , wherein said regulatory regions are functional in the cells of a mammalian host , and optionally , a cationic lipid carrier , and wherein said nucleic acid (nucleic acid) sequence of interest is free of introns .

US8058069B2 Filed: 2008-04-15 Issued: 2011-11-15 Lipid formulations for nucleic acid delivery (Original Assignee) Protiva Biotherapeutics Inc (Current Assignee) Arbutus Biopharma Corp Edward Yaworski, Kieu Lam, Lloyd Jeffs, Lorne Palmer, Ian Maclachlan	WO9324641A2 Filed: 1993-06-02 Issued: 1993-12-09 Adeno-associated virus with inverted terminal repeat sequences as promoter (Original Assignee) The United States Of America, As Represented By The Secretary, Department Of Health & Human Services Barrie J. Carter, Terrence Flotte, Sandra Afione, Rikki Solow
US8058069B2 CLAIM 1 . A nucleic acid-lipid particle comprising : (a) a nucleic acid ; (b) a cationic lipid comprising from 50 mol % to 65 mol % of the total lipid present in the particle ; (c) a non-cationic lipid comprising a mixture of a phospholipid and cholesterol or a derivative thereof , wherein the phospholipid comprises from 4 mol % to 10 mol % of the total lipid present in the particle and the cholesterol or derivative thereof comprises from 30 mol % to 40 mol % of the total lipid present in the particle ; and (d) a conjugated lipid that inhibits aggregation (isolated nucleic acid) of particles comprising from 0 . 5 mol % to 2 mol % of the total lipid present in the particle .	WO9324641A2 CLAIM 12 . An isolated nucleic acid (inhibits aggregation) consisting essentially of the inverted terminal repeat sequences of adeno-associated virus .
US8058069B2 CLAIM 9 . The nucleic acid-lipid particle of claim 1 , wherein the phospholipid comprises dipalmitoylphosphatidylcholine (repeat sequences) (DPPC) , distearoylphosphatidylcholine (DSPC) , or a mixture thereof .	WO9324641A2 CLAIM 1 . An adeno-associated viral vector comprising the inverted terminal repeat sequences (phospholipid comprises dipalmitoylphosphatidylcholine) of adeno-associated virus and a nucleic acid , wherein the inverted terminal repeat sequences promote expression of the nucleic acid in the absence of another promoter .
US8058069B2 CLAIM 10 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation (isolated nucleic acid) of particles comprises a polyethyleneglycol (PEG)-lipid conjugate .	WO9324641A2 CLAIM 12 . An isolated nucleic acid (inhibits aggregation) consisting essentially of the inverted terminal repeat sequences of adeno-associated virus .
US8058069B2 CLAIM 15 . The nucleic acid-lipid particle of claim 1 , wherein the conjugated lipid that inhibits aggregation (isolated nucleic acid) of particles comprises from 1 mol % to 2 mol % of the total lipid present in the particle .	WO9324641A2 CLAIM 12 . An isolated nucleic acid (inhibits aggregation) consisting essentially of the inverted terminal repeat sequences of adeno-associated virus .