Extrahepatic delivery iRNA compositions and methods of use thereof

Abstract

The present invention provides double-stranded ribonucleic acid (dsRNA) agents for inhibiting expression of a target gene comprising an antisense strand that is complementary to the target gene; a sense strand that is complementary to the antisense strand and forms a duplex region with the antisense strand; and one or more C22 hydrocarbon chains conjugated to one or more internal positions of at least one of the strands, compositions comprising such dsRNA agents, and methods of their use to treat subjects having skeletal muscle disorders, cardiac muscle disorders, or adipose tissue disorders.

Claims

1. A double-stranded ribonucleic acid (dsRNA) agent or a pharmaceutically acceptable salt thereof for inhibiting expression of a target gene, an antisense strand that is complementary to the target gene; a sense strand that is complementary to the antisense strand and forms a duplex region with the antisense strand; and C conjugated to position 6 of the sense strand, counting from the 5' end of the sense strand ２２ hydrocarbon chain or a pharmaceutically acceptable salt thereof.

2. C ２２ 10. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, wherein the hydrocarbon chain is saturated or unsaturated.

3. C ２２ 10. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, wherein the hydrocarbon chain is linear or branched.

4. The dsRNA agent or a pharmaceutically acceptable salt thereof of claim 1, wherein the C22 hydrocarbon chain is a linear C22 alkyl chain.

5. The dsRNA agent of claim 4, or a pharmaceutically acceptable salt thereof, wherein the C22 hydrocarbon chain is directly attached to the 2'-O of the sugar moiety at position 6 of the sense strand, counting from the 5'-end of the sense strand.

6. 2. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, wherein the sense strand and the antisense strand are each independently 15 to 30 nucleotides in length; 19 to 25 nucleotides in length; or 21 to 23 nucleotides in length.

7. C ２２ The hydrocarbon chain is C ２２ 10. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, which is an acid.

8. C ２２ 8. The dsRNA agent of claim 7, or a pharmaceutically acceptable salt thereof, wherein the acid is selected from the group consisting of docosanoic acid, 6-octyltetradecanoic acid, 10-hexylhexadecanoic acid, all-cis-7,10,13,16,19-docosapentaenoic acid, all-cis-4,7,10,13,16,19-docosahexaenoic acid, all-cis-13,16-docosadienoic acid, all-cis-7,10,13,16-docosatetraenoic acid, all-cis-4,7,10,13,16-docosapentaenoic acid, and cis-13-docosaenoic acid.

9. C ２２ The hydrocarbon chain is C ２２ 10. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, which is an alcohol.

10. C ２２ 10. The dsRNA agent of claim 9, or a pharmaceutically acceptable salt thereof, wherein the alcohol is selected from the group consisting of 1-docosanol, 6-octyltetradecan-1-ol, 10-hexylhexadecan-1-ol, cis-13-docosene-1-ol, docosan-9-ol, docosan-2-ol, docosan-10-ol, docosan-11-ol, and cis-4,7,10,13,16,19-docosahexanol.

11. C ２２ The hydrocarbon chain is C ２２ 10. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, which is an amide.

12. C ２２ 12. The dsRNA agent of claim 11, or a pharmaceutically acceptable salt thereof, wherein the amide is selected from the group consisting of (E)-Docos-4-enamide, (E)-Docos-5-enamide, (Z)-Docos-9-enamide, (E)-Docos-11-enamide, 12-docosenamid, (Z)-Docos-13-enamide, (Z)-N-hydroxy-13-docosenamid, (E)-Docos-14-enamide, 6-cis-docosenamid, 14-docosenamid Docos-11-enamide, (4E,13E)-Docosa-4,13-dienamide, and (5E,13E)-Docosa-5,13-dienamide.

13. C ２２ 10. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, wherein the hydrocarbon chain is conjugated via a carrier.

14. 14. The dsRNA agent of claim 13, or a pharmaceutically acceptable salt thereof, wherein the carrier is a cyclic group selected from the group consisting of pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, [1,3]dioxolanyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydrofuranyl, and decalinyl; or an acyclic moiety based on a serinol or diethanolamine backbone.

15. C ２２ 2. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, wherein the hydrocarbon chain is conjugated to the dsRNA agent via a linker that contains an ether, a thioether, a urea, a carbonate, an amine, an amide, a maleimide thioether, a disulfide, a phosphodiester, a sulfonamide linkage, a product of a click reaction, or a carbamate.

16. C ２２ 10. The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1, wherein the hydrocarbon chain is conjugated to the dsRNA agent, or a pharmaceutically acceptable salt thereof, via a linker or carrier, or via an internucleotide phosphate linkage.

17. C ２２ 10. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, wherein the hydrocarbon chain is conjugated to a nucleobase, a sugar moiety, or an internucleoside phosphate linkage.

18. 10. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, further comprising a phosphate or phosphate mimetic at the 5'-end of the antisense strand.

19. 19. The dsRNA agent of claim 18, or a pharmaceutically acceptable salt thereof, wherein the phosphomimetic is 5'-vinylphosphonate (VP).

20. 10. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, further comprising a modified phosphate at the 5'-end of the antisense strand.

21. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, further comprising a targeting ligand.

22. C ２２ 2. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, wherein the hydrocarbon chain or the targeting ligand is conjugated via a bio-cleavable linker selected from the group consisting of a DNA linker, an RNA linker, a disulfide linker, an amide linker, a protease-cleavable peptide linker, a functionalized monosaccharide linker, a galactosamine oligosaccharide linker, a glucosamine linker, a glucose linker, a galactose linker, a mannose linker, and combinations thereof.

23. The dsRNA agent or a pharmaceutically acceptable salt thereof according to claim 1, which does not contain an N-acetylgalactosamine (GalNAc) derivative.

24. 2. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, wherein the 3'-end of the sense strand is protected via an end cap that is a cyclic group having an amine, and the cyclic group is selected from the group consisting of pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, [1,3]dioxolanyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, tetrahydrofuranyl, and decalinyl.

25. The target genes are adrenoceptor beta 1 (ADRB1); calcium voltage-dependent channel subunit alpha 1C (CACNA1C); calcium voltage-dependent channel subunit alpha 1G (CACNA1G) (T-type calcium channel); angiotensin II 2. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of: sodium voltage-gated channel alpha subunit 2 (SCN2A); hyperpolarization-activated cyclic nucleotide-gated potassium channel 1 (HCN1); hyperpolarization-activated cyclic nucleotide-gated potassium channel 4 (HCN4); hyperpolarization-activated cyclic nucleotide-gated potassium channel 3 (HCN3); potassium voltage-gated channel subfamily A member 5 (KCNA5); inward rectifier potassium channel subfamily J member 3 (KCNJ3); inward rectifier potassium channel subfamily J member 4 (KCNJ4); phospholamban (PLN); calcium / calmodulin-dependent protein kinase II delta (CAMK2D); or phosphodiesterase 1 (PDE1).

26. 2. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, wherein the target gene is selected from the group consisting of delta 4-desaturase, sphingolipid 1 (DEGS1); leptin (LEP); folliculin (FLCN); zinc finger protein 423 (ZFP423); cyclin-dependent kinase 6 (CDK6); regulation-associated protein of mTOR complex 1 (RPTOR); mechanistic target of rapamycin kinase (mTOR); forkhead box P1 (FOXP1); phosphodiesterase 3B (PDE3B); and activin A receptor type 1C (ACVR1C).

27. 2. The dsRNA agent of claim 1, or a pharmaceutically acceptable salt thereof, wherein the target gene is selected from the group consisting of myostatin (MSTN); cholinergic receptor nicotinic alpha 1 subunit (CHRNA1); cholinergic receptor nicotinic beta 1 subunit (CHRNB1); cholinergic receptor nicotinic delta subunit (CHRND); cholinergic receptor nicotinic epsilon subunit (CHRNE); cholinergic receptor nicotinic gamma subunit (CHRNG); type XIII collagen alpha 1 chain (COL13A1); docking protein 7 (DOK7); LDL receptor-related protein 4 (LRP4); muscle-associated receptor tyrosine kinase (MUSK); synaptic receptor-associated protein (RAPSN); sodium voltage-dependent channel alpha subunit 4 (SCN4A); and double homeobox 4 (DUX4).

28. 28. A cell comprising the dsRNA agent of any one of claims 1 to 27, or a pharmaceutically acceptable salt thereof.

29. 28. A pharmaceutical composition for inhibiting expression of a target gene, comprising the dsRNA agent of any one of claims 1 to 27, or a pharmaceutically acceptable salt thereof.

30. 28. An in vitro method of inhibiting expression of a target gene in a skeletal muscle cell, a cardiac muscle cell, or an adipocyte, comprising contacting the cell with the dsRNA agent of any one of claims 1-27, or a pharmaceutically acceptable salt thereof.

31. 30. The pharmaceutical composition of claim 29 for treating a subject with a skeletal muscle disorder, a cardiac disorder, or an adipose tissue disorder.

32. 32. The pharmaceutical composition of claim 31, wherein the myocardial disorder is selected from the group consisting of hypertrophic obstructive cardiomyopathy (HOCM), familial hypertrophic cardiomyopathy (FHC); heart failure with preserved ejection fraction (HFPEF); atrial fibrillation (AFIB); ventricular fibrillation (VFIB); angina; myocardial infarction (MI); heart failure or heart failure with reduced ejection fraction (HFREF); supraventricular tachycardia (SVT); hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), arrhythmia, and congestive heart failure (CHF).

33. 32. The pharmaceutical composition of claim 31, wherein the lipid disorder is a metabolic disorder.

34. 34. The pharmaceutical composition of claim 33, wherein the metabolic disorder is selected from the group consisting of carbohydrate metabolism disorders, lipid metabolism disorders, hypertension, cardiovascular disease, and weight disorders.

35. 35. The pharmaceutical composition of claim 34, wherein the metabolic disorder is a body weight disorder and administration of the dsRNA agent inhibits expression of the target gene by no more than 40%, no more than 50%, no more than 60%, or no more than 70%.

36. 32. The pharmaceutical composition of claim 31, wherein the skeletal muscle disorder is selected from the group consisting of myostatin-associated muscle hypertrophy, congenital myasthenic syndrome, and facioscapulohumeral muscular dystrophy (FSHD).

37. 32. The pharmaceutical composition of claim 31, wherein the dsRNA agent is administered subcutaneously to the subject.

38. 32. The pharmaceutical composition of claim 31, wherein the dsRNA agent is administered to the subject intramuscularly.

39. 32. The pharmaceutical composition of claim 31, wherein the dsRNA agent is administered intravenously to the subject.

40. 32. The pharmaceutical composition of claim 31, wherein the dsRNA agent is administered to the subject intraperitoneally.

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