Compounds and methods for modulating apolipoprotein C-III expression for improving a diabetic profile

• US 9,909,124 B2
• Filed: 06/23/2014
• Issued: 03/06/2018
• Est. Priority Date: 06/21/2013
• Status: Active Grant

• Current Assignee: Ionis Pharmaceuticals, Inc.
• Original Assignee: Ionis Pharmaceuticals, Inc.
• Inventors: Crooke, Rosanne M., Alexander, Veronica J.
• Primary Examiner(s): Angell, J. E.
• Application Number:
  US14/900,320
• Publication Number:
  US 20160152974A1
• Time in Patent Office: 1,352 Days
• CPC Class Codes: A61K 31/155 Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• 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 2310/11 Antisense
• 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 2310/3525 MOE, methoxyethoxy

Abstract

Provided herein are improved compounds, compositions and associated methods for reducing expression of ApoCIII mRNA and protein in a subject having, or at risk of having, diabetes. Also provided herein are compounds, compositions and associated methods improving peripheral insulin sensitivity, a lipid profile and a diabetes profile as well as reducing free fatty acids and intramyocellular triglyceride deposition.

Claims

1. A method of improving glucose control of a subject comprising administering a compound comprising an antisense oligonucleotide 12-30 nucleobases in length targeting ApoCIII, wherein the subject has moderately controlled diabetes or uncontrolled diabetes, and is on a stable dose of a glucose lowering agent and, upon administration of the compound, the subject exhibits reduced HbA1c, thereby improving the glucose control of the subject.

2. The method of claim 1, wherein the subject is on a stable dose of metformin.

3. The method of claim 1, wherein the subject has fasting triglyceride levels 200 and 500 mg/dl.

4. The method of claim 1, wherein ApoCIII has a nucleic acid sequence as shown in SEQ ID NO:

1 or SEQ ID NO;

2.

5. The method of claim 4, wherein the antisense oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases of a nucleobase sequence of SEQ ID NO:

4.

6. The method of claim 4, wherein the antisense oligonucleotide consists of a nucleobase sequence of SEQ ID NO:

4.

7. The method of claim 4, wherein the nucleobase sequence of the antisense oligonucleotide is at least 80%, at least 90% or 100% complementary to a nucleobase sequence of SEQ ID NO:

1 or SEQ ID NO;

2.

8. The method of claim 1, wherein the antisense oligonucleotide consists of a single-stranded modified oligonucleotide.

9. The method of claim 1, wherein the antisense oligonucleotide consists of 20 linked nucleosides.

10. The method of claim 1, wherein the antisense oligonucleotide has at least one modified internucleoside linkage, sugar moiety or nucleobase.

11. The method of claim 10, wherein the internucleoside linkage of the antisense oligonucleotide is a phosphorothioate internucleoside linkage, the modified sugar is a bicyclic sugar or 2′

-O-methoxyethyl and the modified nucleobase is a 5-methylcytosine.

12. The method of claim 1, wherein the antisense 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 linked nucleosides;

wherein the gap segment is positioned immediately adjacent to and between the 5′

wing segment and the 3′

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

13. The method of claim 1, wherein the antisense oligonucleotide comprises:

(a) a gap segment consisting of 10 linked deoxynucleosides;

(b) a 5′

wing segment consisting of 5 linked nucleosides;

(c) a 3′

wing segment consisting 5 linked nucleosides;

wherein the gap segment is positioned immediately adjacent to and between the 5′

wing segment and the 3′

wing segment, wherein each nucleoside of each wing segment comprises a 2′

-O-methoxyethyl sugar, wherein each cytosine is a 5-methylcytosine, and wherein each internucleoside linkage is a phosphorothioate linkage.

15. The method of claim 1, wherein the antisense oligonucleotide is parenterally administered.

16. The method of claim 15, wherein the parenteral administration is subcutaneous administration.

17. The method of claim 1, further comprising administration of the glucose lowering agent to the subject.

18. The method of claim 17, wherein the glucose lowering agent is metformin.

19. The method of claim 17, wherein the glucose lowering agent is administered concomitantly or sequentially with the antisense oligonucleotide.

20. The method of claim 1, wherein the antisense oligonucleotide is a salt form.

21. The method of claim 1, wherein the compound is in a composition comprising a pharmaceutically acceptable carrier or diluent.

22. The method of claim 1, wherein the subject has triglyceride levels ≧

500 mg/dl.

23. The method of claim 1, wherein the reduction in HbA1c is a reduction of at least 3% to at least 8% compared to baseline levels of HbA1c.

24. The method of claim 1, wherein the subject has triglyceride levels between 200 and 499 mg/dl.

25. The method of claim 23, wherein the subject has fasting triglyceride levels ≧

200 and ≦

500 mg/dl.

26. The method of claim 1, wherein the subject has triglyceride levels of at least 150 m g/dl.

27. The method of claim 1, wherein the subject has moderately controlled diabetes and an HbA1c level of 7% to 9%.

28. The method of claim 1, wherein the subject has uncontrolled diabetes and an HbA1c level greater than 9%.

29. The method of claim 1, wherein upon administration of the compound, the subject exhibits changes in one or more physiological markers, wherein the changes are selected from among one or more of:

insulin sensitivity index, glucose disposal rate, glucose MCR and/or glucose metabolism;

insulin ration is improved;

free fatty acids, triglycerides, non-HDL-C, VLDL-C, ApoCIII containing VLDL, ApoB and/or LDL-C is reduced; and

HDL-C is increased.

14. A method of improving glucose control in a subject comprising, selecting a subject with moderately controlled diabetes or uncontrolled diabetes, who is on a stable dose of a glucose lowering agent, and administering to the subject a therapeutically effective amount of a compound comprising an antisense oligonucleotide having the sequence of SEQ ID NO:

4 wherein the antisense oligonucleotide comprises;

(a) a gap segment consisting of 10 linked deoxynucleosides;

(b) a 5′

wing segment consisting of 5 linked nucleosides;

(c) a 3′

wing segment consisting 5 linked nucleosides;

wherein the gap segment is positioned immediately adjacent to and between the 5′

wing segment and the 3′

wing segment, wherein each nucleoside of each wing segment comprises a 2′

-O-methoxyethyl sugar, wherein each cytosine is a 5-methylcytosine, wherein each internucleoside linkage is a phosphorothioate linkage, and wherein upon administration of the compound, the subject exhibits reduced HbA1c, thereby improving the glucose control in the subject.