MODULATION OF ANGIOPOIETIN-LIKE 3 EXPRESSION

• US 20130023579A1
• Filed: 01/07/2011
• Published: 01/24/2013
• Est. Priority Date: 01/08/2010
• Status: Active Grant

• Current Assignee: Ionis Pharmaceuticals, Inc.
• Original Assignee: ISIS Pharmaceuticals Incorporated
• Inventors: Crooke, Rosanne M., Graham, Mark J., Dobie, Kenneth W., Lee, Richard
• Granted Patent: US 8,653,047 B2
• Time in Patent Office: Days
• US Class Current: 514/44.A
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/08 for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 Antihypertensives
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/1136 against growth factors, growth regulators, cytokines, lymphokines or hormones
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3231 having an additional ring, e.g. LNA, ENA
• CPC Class Codes: C12N 2310/3341 5-Methylcytosine
• CPC Class Codes: C12N 2310/341 Gapmers, i.e. of the type ===---===
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 2320/30 : Special therapeutic applications
• CPC Class Codes: C12Q 1/6883 : for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/136 : Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/158 : Expression markers
• CPC Class Codes: G01N 33/5023 : on expression patterns

Abstract

Provided herein are methods, compounds, and compositions for reducing expression of an ANGPTL3 mRNA and protein in an animal. Also provided herein are methods, compounds, and compositions for reducing plasma lipids, plasma glucose and atherosclerotic plaques in an animal. Such methods, compounds, and compositions are useful to treat, prevent, delay, or ameliorate any one or more of cardiovascular disease or metabolic disease, or a symptom thereof.

Claims

1. A method of reducing ANGPTL3 expression in an animal comprising administering to the animal a compound comprising a modified oligonucleotide 10 to 30 linked nucleosides in length targeted to ANGPTL3, wherein expression of ANGPTL3 is reduced by at least 40% in the animal.

2. The method of claim 1, wherein reducing ANGPT3 expression in the animal

(a) reduces apoC-III expression levels;

(b) reduces triglyceride levels;

(c) reduces cholesterol levels;

(d) reduces LDL levels;

(e) reduces glucose levels;

(f) improves insulin sensitivity; and

/or

(g) ameliorates a metabolic or cardiovascular disease.

8. The method of claim 1, wherein the modified oligonucleotide has a nucleobase sequence at least 90%, at least 95% or 100% complementary to any of SEQ ID NO:

1-5 as measured over the entirety of said modified oligonucleotide.

9. The method of claim 1, wherein the modified oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases of sequence recited in any one of SEQ ID NOs:

34, 37-38, 40-43, 45, 50-55, 57-64, 67, 69-75, 77-78, 81-83, 85, 87, 90, 92-103, 105-107, 109-110, 113-115, 117-123, 126-146, 148-160, 162-180.

10. The method of claim 1, wherein the animal is a human.

11. The method of claim 1, wherein the compound is a first agent and further comprising administering a second agent.

29. The method of claim 1, wherein at least one internucleoside linkage of said modified oligonucleotide is a modified internucleoside linkage, at least one nucleoside of said modified oligonucleotide comprises a modified sugar and/or at least one nucleoside of said modified oligonucleotide comprises a modified nucleobase.

30. The method of claim 29, wherein each internucleoside linkage is a phosphorothioate internucleoside linkage.

34. The method of claim 29, wherein at least one modified sugar is a bicyclic sugar.

35. The method of claim 29, wherein at least one modified sugar comprises a 2′

-O-methoxyethyl, a (4′

-CH(CH₃)—

O-2′

) BNA, a (4′

-CH₂—

O-2′

) BNA, (4′

-CH₂—

O-2′

) BNA or a 4′

-(CH₂)_n—

O-2′

bridge, wherein n is 1 or 2.

37. The method of claim 29, wherein the modified nucleobase is a 5-methylcytosine.

38. The method of claim 1, wherein the modified oligonucleotide consists of 20 linked nucleosides.

39. The method of claim 1, wherein the modified oligonucleotide comprises:

a. a gap segment consisting of linked deoxynucleosides;

b. a 5′

wing segment consisting of linked nucleosides;

c. a 3′

wing segment consisting of linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

49. The method of claim 2, wherein the levels are independently reduced by 5%, 10%, 20%, 30%, 35%, or 40%.

3-7. -7. (canceled)

12-28. -28. (canceled)

31-33. -33. (canceled)

36. (canceled)

40. (canceled)

41. A method for treating an animal with metabolic or cardiovascular disease comprising

a. identifying said animal with metabolic or cardiovascular disease,

b. administering to said animal a therapeutically effective amount of a compound comprising a modified oligonucleotide consisting of 20 linked nucleosides and having a nucleobase sequence comprising at least 8 contiguous nucleobases of a nucleobase sequence selected from any of SEQ ID NO:

34-182,

wherein said animal with metabolic or cardiovascular disease is treated.

42-48. -48. (canceled)

50-53. -53. (canceled)

54. A compound comprising a modified oligonucleotide consisting of 10 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8 contiguous nucleobases of a nucleobase sequence selected from any of SEQ ID NO:

34-182.

55. The compound of claim 54, wherein the nucleobase sequence of the modified oligonucleotide is at least 90%, at least 95% or 100% complementary to any of SEQ ID NOs:

1-5.

58. The compound of claim 54, wherein at least one internucleoside linkage is a modified internucleoside linkage, at least one nucleoside comprises a modified sugar and/or at least one nucleoside comprises a modified nucleobase.

59. The compound of claim 58, wherein each internucleoside linkage is a phosphorothioate internucleoside linkage.

61. The compound of claim 58, wherein at least one modified sugar is a bicyclic sugar.

62. The compound of claim 58, wherein at least one modified sugar comprises a 2′

-O-methoxyethyl, a (4′

-CH(CH₃)—

O-2′

) BNA, a (4′

-C_1-17—

O-2′

) BNA, (4′

-CH₂—

O-2′

) BNA or a 4′

-(CH₂)_n—

O-2′

bridge, wherein n is 1 or 2.

64. The compound of claim 58, wherein the modified nucleobase is a 5-methylcytosine.

56-57. -57. (canceled)

60. (canceled)

63. (canceled)

65-71. -71. (canceled)