Composition and method for suppressing the expression of LPA (APO(A)) protein

Claims

1. A double-stranded ribonucleic acid (dsRNA)-based agent that suppresses the expression of LPA (Apo(a)), wherein the dsRNA-based agent comprises a sense strand and an antisense strand and optionally comprises a targeting ligand; a region complementary to the LPA RNA transcript is contained at nucleotide positions 2-18 in the antisense strand, wherein the complementary region comprises at least 15 consecutive nucleotides that differ by 0, 1, 2, or 3 nucleotides from one of the antisense sequences set forth in one of Tables 1-3.

2. The dsRNA-based agent according to claim 1, wherein the region complementary to the LPA RNA transcript comprises at least 15, 16, 17, 18 or 19 consecutive nucleotides that differ by no more than 3 nucleotides from one of the antisense sequences indicated in one of Tables 1-3.

3. The dsRNA-based agent according to claim 1 or claim 2, wherein the antisense strand of the dsRNA is at least substantially complementary to any target region in the mRNA of the human LPA gene and is represented in any of Tables 1-3.

4. The dsRNA-based agent of claim 3, wherein the antisense strand of the dsRNA is fully complementary to any target region in the mRNA of the human LPA gene and is represented in any of Tables 1-3.

5. The dsRNA-based agent of claim 1, wherein the dsRNA-based agent comprises a sense strand sequence set forth in any one of Tables 1-3, wherein the sense strand sequence is at least substantially complementary to a sequence of the antisense strand in the dsRNA-based agent.

6. The dsRNA-based agent of claim 1, wherein the dsRNA-based agent comprises a sense strand sequence set forth in any of Tables 1-3, wherein the sense strand sequence is fully complementary to the antisense strand sequence in the dsRNA-based agent.

7. The dsRNA-based agent of claim 1, wherein the dsRNA-based agent comprises an antisense strand sequence specified in any of Tables 1-3.

8. The dsRNA-based agent of claim 1, wherein the dsRNA-based agent comprises a sequence indicated as a duplex sequence in any of Tables 1-3.

9. The dsRNA-based agent of claim 1, wherein the dsRNA-based agent comprises at least one modified nucleotide.

10. The dsRNA-based agent of claim 1, wherein all or substantially all of the nucleotides in the antisense strand are modified nucleotides.

11. The dsRNA-based agent of claim 5 or 6, wherein the at least one modified nucleotide comprises a 2'-O-methyl nucleotide, a 2'-fluoronucleotide, a 2'-deoxynucleotide, a 2',3'-seconucleotide mimetic, a locked nucleotide, an unlocked nucleic acid (UNA) nucleotide, a glycol nucleic acid (GNA) nucleotide, a 2'-F-arabinose nucleotide, a 2'-methoxyethyl nucleotide, an anhydrous nucleotide, a ribitol, a reverse nucleotide, a reverse anhydrous nucleotide, a reverse 2'-OMe nucleotide, a reverse 2'-deoxynucleotide, a 2'-amino modified nucleotide, a 2'-alkyl modified nucleotide, a morpholino nucleotide and a 3'-OMe nucleotide, a nucleotide containing a 5'-phosphorothioate group, or a terminal nucleotide linked to a cholesterol derivative or a bisdecylamide group of dodecanoic acid, a 2'-amino modified nucleotide, a phosphoramidate, or a nucleotide containing an unnatural base.

12. The dsRNA-based agent according to claim 9 or 10, wherein the E-vinylphosphonate nucleotide is contained at the 5' end of the guide strand.

13. The dsRNA-based agent of claim 1, wherein the dsRNA-based agent comprises at least one phosphorothioate internucleoside linkage.

14. The dsRNA-based agent according to claim 1, wherein the sense strand comprises at least one phosphorothioate internucleoside linkage.

15. The dsRNA-based agent of claim 1, wherein the antisense strand comprises at least one phosphorothioate internucleoside linkage.

16. The dsRNA-based agent according to claim 1, wherein the sense strand comprises 1, 2, 3, 4, 5 or 6 phosphorothioate internucleoside linkages.

17. The dsRNA-based agent of claim 1, wherein the antisense strand comprises 1, 2, 3, 4, 5 or 6 phosphorothioate internucleoside linkages.

18. The dsRNA-based agent of claim 1, wherein all or substantially all nucleotides of the sense strand and the antisense strand are modified nucleotides.

19. The dsRNA-based agent of claim 1, wherein the modified sense strand is a modified sense strand sequence specified in one of Tables 2-3.

20. The dsRNA-based agent of claim 1, wherein the modified antisense strand is a modified antisense strand sequence specified in one of Tables 2-3.

21. The dsRNA-based agent according to claim 1, wherein the sense strand is complementary or substantially complementary to the antisense strand, and the length of the complementary region is from 16 to 23 nucleotides.

22. The dsRNA-based agent according to claim 21, wherein the length of the complementary region is from 19 to 21 nucleotides.

23. The dsRNA-based agent according to claim 1, wherein the length of each strand is no more than 30 nucleotides.

24. The dsRNA-based agent according to claim 1, wherein the length of each strand is no more than 25 nucleotides.

25. The dsRNA-based agent according to claim 1, wherein the length of each strand is no more than 23 nucleotides.

26. The dsRNA-based agent of claim 1, wherein the dsRNA-based agent comprises at least one modified nucleotide and further comprises one or more targeting groups or linking groups.

27. The dsRNA-based agent of claim 26, wherein one or more targeting groups or linking groups are conjugated to the sense strand.

28. The dsRNA-based agent according to claim 26 or 27, wherein the targeting groups or linking groups comprise N-acetylgalactosamine (GalNAc).

29. The dsRNA-based agent according to claim 26 or 27, wherein the targeting groups have the following structures:

30. The dsRNA-based agent of claim 1, wherein the dsRNA-based agent comprises a targeting group conjugated to the 5' end of the sense strand.

31. The dsRNA-based agent of claim 1, wherein the dsRNA-based agent comprises a targeting group conjugated to the 3' end of the sense strand.

32. The dsRNA-based agent of claim 1, wherein the antisense strand comprises one reverse abasic residue at the 3' end.

33. The dsRNA-based agent of claim 1, wherein the sense strand comprises one or two reverse basic residues at the 3' end and / or 5' end, or the sense strand comprises one or two isomannite residues at the 3' end and / or 5' end.

34. The dsRNA-based agent of claim 1, wherein the dsRNA-based agent has two blunt ends.

35. The dsRNA-based agent according to claim 1, wherein at least one strand comprises a 3'-protruding region of at least 1 nucleotide.

36. The dsRNA-based agent according to claim 1, wherein at least one strand comprises a 3'-protruding region of at least 2 nucleotides.

37. A composition comprising a dsRNA-based agent according to any one of claims 1-36.

38. The composition according to claim 37, additionally containing a pharmaceutically acceptable carrier.

39. The composition according to claim 38, further comprising one or more additional therapeutic agents.

40. The composition of claim 39, wherein the composition is packaged in a kit, container, package, dispenser, pre-filled syringe or vial.

41. The composition of claim 37, wherein the composition is formulated for subcutaneous administration or is formulated for intravenous (IV) administration.

42. A cell comprising a dsRNA-based agent according to any one of claims 1-36.

43. The cell of claim 42, wherein the cell is a mammalian cell, not necessarily a human cell.

44. A method for suppressing the expression of the LPA gene in a cell, comprising (i) obtaining a cell comprising an effective amount of a double-stranded ribonucleic acid (dsRNA) agent according to any one of claims 1-36 or a composition according to any one of claims 37-41.

45. The method according to claim 44, further comprising (ii) maintaining the cell obtained in (i) of paragraph 44 for a period of time sufficient to cause the destruction of the mRNA transcript of the LPA gene, thereby suppressing the expression of the LPA gene in the cell.

46. The method according to claim 44, wherein the cell is in the body of the subject, and the dsRNA-based agent is administered to the subject subcutaneously.

47. The method of claim 44, wherein the cell is in the subject's body and the dsRNA-based agent is administered to the subject via IV administration.

48. The method of claim 4 6 or 47, further comprising assessing the suppressive effect on the LPA gene after administration of the dsRNA-based agent to the subject, wherein the assessment methods comprise (i) identifying one or more physiological characteristics of an LPA-related disease or condition in a subject and (ii) comparison of the identified physiological characteristics with baseline physiological characteristics prior to treatment of the LPA-related disease or condition and / or control physiological characteristics of the LPA-related disease or condition, wherein the comparison indicates one or more of the presence or absence of suppression of LPA gene expression in the subject.

49. The method according to claim 48, wherein the identified physiological characteristic is the level of Lp(a) in the blood.

50. The method of claim 49, wherein a decrease in the level of Lp(a) in the blood of the subject indicates a decrease in the expression of the LPA gene in the subject.

51. A method for suppressing the expression of the LPA gene in a subject, comprising administering to the subject an effective amount of a double-stranded ribonucleic acid (dsRNA) agent according to any one of claims 1-36 or a composition according to any one of claims 37-41.

52. The method of claim 51, wherein the dsRNA-based agent is administered to the subject subcutaneously.

53. The method of claim 51, wherein the dsRNA-based agent is administered to the subject via IV administration.

54. The method of any one of claims 51-53, further comprising assessing the suppressive effect on the LPA gene after administration of the dsRNA-based agent to the subject, wherein the assessment methods comprise (i) identifying one or more physiological characteristics of an LPA-related disease or condition in a subject and (ii) comparison of the identified physiological characteristics with baseline physiological characteristics before treatment of the LPA-related disease or condition and / or control of physiological characteristics of the LPA-related disease or condition, wherein the comparison indicates one or more of the presence or absence of suppression of LPA gene expression in the subject.

55. The method according to claim 54, wherein the identified physiological characteristic is the level of Lp(a) in the blood.

56. The method of claim 55, wherein a decrease in the level of Lp(a) in the blood of the subject indicates a decrease in the expression of the LPA gene in the subject.

57. A method for treating and preventing an LPA protein-associated disease or condition comprising administering to a subject an effective amount of a double-stranded ribonucleic acid (dsRNA) agent according to any one of claims 1-36 or a composition according to any one of claims 37-41 to suppress the expression of the LPA gene.

58. The method of claim 57, wherein the disease or condition is a cardiovascular disease, wherein the cardiovascular disease comprises Berger's disease, peripheral arterial disease, coronary heart disease, metabolic syndrome, acute coronary syndrome, aortic stenosis, aortic regurgitation, aortic dissection, retinal artery occlusion, cerebrovascular disease, mesenteric thrombosis, superior mesenteric artery occlusion, renal artery stenosis, stable / unstable angina, acute coronary syndrome, heterozygous or homozygous familial hypercholesterolemia, hyperapolipoproteinbetalipoproteinemia, cerebrovascular atherosclerosis, cerebrovascular disease and venous thrombosis, stroke, atherosclerosis, thrombosis, forms of ischemic heart disease or aortic stenosis and / or any other diseases or pathologies associated with elevated levels of Lp(a)-containing particles.

59. The method of claim 57, further comprising prescribing to the subject an additional therapeutic regimen.

60. The method of claim 59, wherein the additional therapeutic regimen comprises administering to the subject one or more LPA antisense polynucleotides of the present invention, administering to the subject a therapeutic agent other than a dsRNA LPA-based agent, and behavioral modifications in the subject.

61. The method of claim 60, wherein the therapeutic agent other than the dsRNA LPA-based agent is one or more additional therapeutic agents such as an HMg-Co-A reductase inhibitor (statins), ezetimibe, a PCSK-9 inhibitor, a STER inhibitor, a therapy targeting ANGPTL3, a therapy targeting AGT, a therapy targeting APOC3, and niacin, or any combination thereof.

62. The method of claim 57, wherein the dsRNA-based agent is administered to the subject subcutaneously.

63. The method of claim 57, wherein the dsRNA-based agent is administered to the subject via IV administration.

64. The method according to any one of paragraphs 57-63, further comprising determining the effectiveness of a double-stranded ribonucleic acid (dsRNA) agent administered to the subject.

65. The method of claim 64, wherein the method for determining the effectiveness of treatment in a subject comprises (i) identifying one or more physiological characteristics of an LPA-related disease or condition in a subject and (ii) comparison of the identified physiological characteristics with the baseline physiological characteristics before treatment of the LPA-related disease or condition, wherein the comparison is indicative of one or more of the presence or absence of efficacy and the level of efficacy of administering to a subject a double-stranded ribonucleic acid (dsRNA)-based agent.

66. The method according to claim 65, wherein the identified physiological characteristic is the level of Lp(a) in the blood.

67. The method according to claim 65, wherein a decrease in the level of Lp(a) in the blood of the subject indicates the effectiveness of administering to the subject a double-stranded ribonucleic acid (dsRNA)-based agent.

68. A method for reducing the level of LPA protein in a subject compared to the subject's baseline level of LPA protein prior to treatment, comprising administering to the subject an effective amount of a double-stranded ribonucleic acid (dsRNA) agent according to any one of claims 1-36 or a composition according to any one of claims 37-41 to reduce the level of expression of the LPA gene.

69. The method of claim 68, wherein the dsRNA-based agent is administered to the subject subcutaneously or administered to the subject by IV administration.

70. A method for altering a physiological characteristic of an LPA-associated disease or condition in a subject compared to a baseline physiological characteristic of an LPA-associated disease or condition in the subject, comprising administering to the subject an effective amount of a double-stranded ribonucleic acid (dsRNA) agent according to any one of claims 1-36 or a composition according to any one of claims 37-41 to alter the physiological characteristic of an LPA-associated disease or condition in the subject.

71. The method of claim 70, wherein the dsRNA-based agent is administered to the subject subcutaneously or administered to the subject by IV administration.

72. The method according to claim 70, wherein the physiological characteristic is the level of Lp(a) in the blood.