# US 8,273,869 B2 · Lipid formulated dsRNA targeting the PCSK9 gene > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,273,869 B2 - Document ID: US8273869B2 - Status: Granted - Kind: B2 - Application number: 12/816,207 - Filed: 2010-06-15 - Published: 2011-01-20 - Granted: 2012-09-25 - Earliest claimed: 2009-06-15 - Assignees: Alnylam Pharmaceuticals, Inc. - Inventors: Kevin Fitzgerald; Gregory Hinkle; Akin Akinc; Stuart Milstein - Web page: https://patentgrep.com/patent/US8273869B2 ## CPC - A61K 31/713: Double-stranded nucleic acids or oligonucleotides - A61K 9/0019: {Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00)} - A61K 9/1272: comprising non-phosphatidyl surfactants as bilayer-forming substances, e.g. cationic lipids or non-phosphatidyl liposomes coated or grafted with polymers (lipids as modifying agents {A61K47/543}) - A61K 9/1273: Polymersomes; Liposomes with polymerisable or polymerised bilayer-forming substances - A61K 38/1709: {from mammals} - A61K 47/545: {Heterocyclic compounds (A61K47/558 takes precedence)} - A61K 47/55: the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug - A61K 47/6911: {the form being a liposome} - A61P 3/06: Antihyperlipidemics - A61P 9/10: for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis - A61P 35/00: Antineoplastic agents - A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 - C12N 15/1137: {against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138)} - C12N 2310/315: Phosphorothioates - C12N 2310/321: 2'-O-R Modification - C12N 2310/322: 2'-R Modification - C12N 2310/3515: Lipophilic moiety, e.g. cholesterol - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract This invention relates to composition and methods using lipid formulated siRNA targeted to a PCSK9 gene. ## Claims ### Claim 1 A composition comprising a nucleic acid lipid particle comprising a double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of a human PCSK9 gene in a cell, wherein: the nucleic acid lipid particle comprises a lipid formulation comprising 45-65 mol % of a cationic lipid, 5 mol % to about 10 mol %, of a non-cationic lipid, 25-40 mol % of a sterol, and 0.5-5 mol % of a PEG or PEG-modified lipid and the cationic lipid comprises MC3 (((6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate), the dsRNA consists of a sense strand and an antisense strand, wherein the sense strand consists of the nucleotide sequence of SEQ ID NO:1227 and the antisense strand consists of the nucleotide sequence of SEQ ID NO:1228. ### Claim 2 (depends on 1) The composition of claim 1, wherein each strand is modified as follows to include a 2′-O-methyl ribonucleotide as indicated by a lower case letter “c” or “u” and a phosphorothioate as indicated by a lower case letter “s”: the dsRNA consists of a sense strand consisting of SEQ ID NO:1229 (5′-uucuAGAccuGuuuuGcuuTsT-3′) and an antisense strand consisting of SEQ ID NO:1230 (5′-AAGcAAAAcAGGUCuAGAATsT-3′). ### Claim 3 (depends on 1, 2) The composition of claim 1 or 2, wherein the lipid formulation is selected from the group consisting of: LNP11 MC3/DSPC/Cholesterol/PEG-DMG 50/10/38.5/1.5 LNP14 MC3/DSPC/Cholesterol/PEG-DMG 40/15/40/5 LNP15 MC3/DSPC/Cholesterol/PEG-DSG/GalNAc-PEG-DSG 50/10/35/4.5/0.5 LNP16 MC3/DSPC/Cholesterol/PEG-DMG 50/10/38.5/1.5 LNP17 MC3/DSPC/Cholesterol/PEG-DSG 50/10/38.5/1.5 LNP18 MC3/DSPC/Cholesterol/PEG-DMG 50/10/38.5/1.5 LNP19 MC3/DSPC/Cholesterol/PEG-DMG 50/10/35/5 LNP20 MC3/DSPC/Cholesterol/PEG-DPG 50/10/38.5/1.5. ### Claim 4 (depends on 1, 2) The composition of claim 1 or claim 2, further comprising a lipoprotein. ### Claim 5 (depends on 1, 2) The composition of claim 1 or claim 2, further comprising apolipoprotein E (ApoE). ### Claim 6 (depends on 5) The composition of claim 5, wherein the dsRNA is conjugated to a lipophile. ### Claim 7 (depends on 6) The composition of claim 6, wherein the lipophile is cholesterol. ### Claim 8 (depends on 5) The composition of claim 5, wherein the ApoE is reconstituted with at least one amphiphilic agent. ### Claim 9 (depends on 8) The composition of claim 8, wherein the amphiphilic agent is a phospholipid. ### Claim 10 (depends on 8) The composition of claim 8, wherein the amphilic agent is a phospholipid selected from the group consisting of dimyristoyl phosphatidyl choline (DMPC), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), egg phosphatidylcholine (EPC), distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), -phosphatidylethanolamine (POPE), dioleoyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), and combinations thereof. ### Claim 11 (depends on 9) The composition of claim 9, wherein the ApoE is reconstituted high density lipoprotein (rHDL). ### Claim 12 (depends on 3) The composition of claim 3, further comprising a lipoprotein. ### Claim 13 (depends on 3) The composition of claim 3, further comprising apolipoprotein E (ApoE). ### Claim 14 (depends on 13) The composition of claim 13, wherein the dsRNA is conjugated to a lipophile. ### Claim 15 (depends on 14) The composition of claim 14, wherein the lipophile is cholesterol. ### Claim 16 (depends on 15) The composition of claim 15, wherein the ApoE is reconstituted with at least one amphiphilic agent. ### Claim 17 (depends on 16) The composition of claim 16, wherein the amphiphilic agent is a phospholipid. ### Claim 18 (depends on 16) The composition of claim 16, wherein the amphilic agent is a phospholipid selected from the group consisting of dimyristoyl phosphatidyl choline (DMPC), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), egg phosphatidylcholine (EPC), distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), -phosphatidylethanolamine (POPE), dioleoyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), and combinations thereof. ### Claim 19 (depends on 13) The composition of claim 13, wherein the ApoE is reconstituted high density lipoprotein (rHDL). ### Claim 20 (depends on 3) The composition of claim 3, wherein the lipid formulation is LNP11. ### Claim 21 (depends on 3) The composition of claim 3, wherein the lipid formulation is LNP14. ### Claim 22 (depends on 3) The composition of claim 3, wherein the lipid formulation is LNP15. ### Claim 23 (depends on 3) The composition of claim 3, wherein the lipid formulation is LNP16. ### Claim 24 (depends on 3) The composition of claim 3, wherein the lipid formulation is LNP17. ### Claim 25 (depends on 3) The composition of claim 3, wherein the lipid formulation is LNP18. ### Claim 26 (depends on 3) The composition of claim 3, wherein the lipid formulation is LNP19. ### Claim 27 (depends on 3) The composition of claim 3, wherein the lipid formulation is LNP20.