Compositions and methods for modulating growth hormone receptor expression

• US 9,994,855 B2
• Filed: 05/01/2015
• Issued: 06/12/2018
• Est. Priority Date: 05/01/2014
• Status: Active Grant

• Current Assignee: Ionis Pharmaceuticals, Inc.
• Original Assignee: Ionis Pharmaceuticals, Inc.
• Inventors: Prakash, Thazha P., Seth, Punit P., Swayze, Eric E., Bhanot, Sanjay, Freier, Susan M., Bui, Huynh-Hoa
• Primary Examiner(s): Schnizer, Richard A
• Application Number:
  US15/307,990
• Publication Number:
  US 20170166899A1
• Time in Patent Office: 1,138 Days
• Field of Search: None
• CPC Class Codes: A61K 47/549 Sugars, nucleosides, nucleotides or nucleic acids
• CPC Class Codes: A61K 47/56 the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/06 of the anterior pituitary hormones, e.g. TSH, ACTH, FSH, LH, PRL, GH
• CPC Class Codes: C12N 15/1138 against receptors or cell surface proteins
• 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 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/351 Conjugate
• CPC Class Codes: C12N 2310/3525 MOE, methoxyethoxy

Abstract

The present embodiments provide methods, compounds, and compositions for treating, preventing, ameliorating a disease associated with excess growth hormone using antisense compounds oligonucleotides targeted to growth hormone receptor (GHR).

Claims

1. A compound comprising a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide is 15 to 30 linked nucleosides in length and wherein the modified oligonucleotide has a nucleobase sequence comprising a portion of at least 15 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 153921-153940 of SEQ ID NO:

2.

2. The compound of claim 1, wherein the modified oligonucleotide comprises at least one modification selected from at least one modified internucleoside linkage, at least one modified sugar, and at least one modified nucleobase.

3. The compound of claim 2, wherein the modified internucleoside linkage is a phosphorothioate internucleoside linkage.

4. The compound of claim 2, wherein the modified oligonucleotide comprises at least 1 phosphodiester internucleoside linkage.

5. The compound of claim 2, wherein the modified sugar is a bicyclic sugar.

6. The compound of claim 5, wherein the bicyclic sugar is selected from the group consisting of:

4′

-(CH₂)—

O-2′

(LNA);

4′

-(CH₂)₂—

O-2′

(ENA); and

4′

-CH(CH₃)—

O-2′

(cEt).

7. The compound of claim 2, wherein the modified sugar is 2′

-O-methoxyethyl.

8. The compound of claim 2, wherein the modified nucleobase is a 5-methylcytosine.

9. The compound of claim 1, wherein the conjugate group comprises:

10. The compound of claim 1, wherein the conjugate group comprises:

11. The compound of claim 1, wherein the conjugate group comprises:

12. The compound of claim 1, wherein the conjugate group comprises at least one phosphorus linking group or neutral linking group.

13. The compound of claim 1, wherein the conjugate group comprises a cell-targeting moiety.

14. The compound of claim 13, wherein the cell-targeting moiety comprises:

15. The compound of claim 13, wherein the cell-targeting moiety comprises:

16. The compound of claim 13, wherein the cell-targeting moiety comprises:

17. The compound of claim 1, wherein the conjugate group comprises a cleavable moiety selected from among:

a phosphodiester, an amide, or an ester.

18. The compound of claim 1, wherein the conjugate group comprises a phosphodiester cleavable moiety.

19. The compound of claim 1, wherein the conjugate group does not comprise a cleavable moiety, and wherein the conjugate group comprises a phosphorothioate linkage between the conjugate group and the oligonucleotide.

20. The compound of claim 1, wherein the conjugate group comprises an amide cleavable moiety.

21. The compound of claim 1, wherein the conjugate group comprises an ester cleavable moiety.

24. A composition comprising the compound of claim 1 and at least one of a pharmaceutically acceptable carrier or diluent.

25. A prodrug comprising the compound of claim 1.

27. A method comprising administering to an animal the compound or composition of claim 24.

28. The method of claim 27, wherein the animal is a human.

29. The method of claim 28, wherein the human has a disease associated with excess growth hormone.

30. The method of claim 29, wherein the disease associated with excess growth hormone is acromegaly.

31. The method of claim 29, wherein the administration reduces IGF-1 levels.

32. A method of reducing growth hormone receptor (GHR) levels in a human comprising administering to the human a therapeutically effective amount of the compound of claim 1, thereby reducing GHR levels in the human.

33. The method of claim 32, wherein the human has a disease associated with excess growth hormone.

34. The method of claim 33, wherein the disease associated with excess growth hormone is acromegaly.

36. The composition of claim 24, wherein the compound is a salt thereof.

37. The compound of claim 1, wherein the modified oligonucleotide is 16 to 30 linked nucleosides in length and wherein the modified oligonucleotide has a nucleobase sequence comprising a portion of at least 16 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 153921-153940 of SEQ ID NO:

2.

38. The compound of claim 1, wherein the modified oligonucleotide is 17 to 30 linked nucleosides in length and wherein the modified oligonucleotide has a nucleobase sequence comprising a portion of at least 17 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 153921-153940 of SEQ ID NO:

2.

39. The compound of claim 1, wherein the modified oligonucleotide is 18 to 30 linked nucleosides in length and wherein the modified oligonucleotide has a nucleobase sequence comprising a portion of at least 18 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 153921-153940 of SEQ ID NO:

2.

40. The compound of claim 1, wherein the modified oligonucleotide is 19 to 30 linked nucleosides in length and wherein the modified oligonucleotide has a nucleobase sequence comprising a portion of at least 19 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 153921-153940 of SEQ ID NO:

2.

41. The compound of claim 1, wherein the modified oligonucleotide is 20 to 30 linked nucleosides in length and wherein the modified oligonucleotide has a nucleobase sequence comprising 20 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 153921-153940 of SEQ ID NO:

2.

22. A compound consisting of

(a) a conjugate group and

(b) a modified oligonucleotide of nucleobases of SEQ ID NO:

703 according to the following formula;

mCes mCes Aes mCes mCes Tds Tds Tds Gds Gds Gds Tds Gds Ads Ads Tes Aes Ges mCes Ae;

wherein,

A=an adenine,

mC=a 5-methylcytosine

G=a guanine,

T=a thymine,

e=a 2′

-O-methoxyethyl modified nucleoside,

d=a 2′

-deoxynucleoside, and

s=a phosphorothioate internucleoside linkage.

23. A compound consisting of a conjugate group and a modified oligonucleotide of nucleobases of SEQ ID NO:

703 according to the following formula;

26. A composition comprising the compound of claim 23 and at least one of a pharmaceutically acceptable carrier or diluent.

35. A compound consisting of a modified oligonucleotide and a conjugate group, wherein the modified oligonucleotide is 20 linked nucleosides in length and consists of the sequence of SEQ ID NO:

703, wherein the modified oligonucleotide consists of

a gap segment consisting of ten linked deoxynucleosides;

a 5′

wing segment consisting of five linked nucleosides; and

a 3′

wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

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

-MOE sugar, wherein each internucleoside linkage is a phosphorothioate linkage;

wherein each cytosine is a 5-methylcytosine; and

wherein the conjugate group is

42. The compound of claim 35, wherein the compound is a salt thereof.

43. A compound comprising a modified oligonucleotide, wherein the modified oligonucleotide is 20 linked nucleosides in length and consists of the sequence of SEQ ID NO:

703, wherein the modified oligonucleotide consists of

a gap segment consisting of ten linked deoxynucleosides;

a 5′

wing segment consisting of five linked nucleosides; and

a 3′

wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

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

-MOE sugar, wherein each internucleoside linkage is a phosphorothioate linkage; and

wherein each cytosine is a 5-methylcytosine.