Modified relaxin polypeptides comprising a pharmacokinetic enhancer and uses thereof

• US 10,266,578 B2
• Filed: 02/08/2018
• Issued: 04/23/2019
• Est. Priority Date: 02/08/2017
• Status: Active Grant

• Current Assignee: Bristol-Myers Squibb Company
• Original Assignee: Bristol-Myers Squibb Company
• Inventors: Dubowchik, Gene M., Gudmundsson, Olafur S., Han, Xiaojun, Lawrence, R. Michael, Lipovsek, Dasa, Madsen, Cort S., Mapelli, Claudio, Morin, Paul E., Myers, Michael C.
• Primary Examiner(s): Saoud, Christine J
• Application Number:
  US15/891,492
• Publication Number:
  US 20180222960A1
• Time in Patent Office: 439 Days
• Field of Search: None
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 38/18 Growth factors; Growth regulators
• CPC Class Codes: A61K 38/2221 Relaxins
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 47/64 Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent peptidic linkers A61K47/65
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 27/12 for cataracts
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• 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 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/06 : Antiarrhythmics
• 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: C07K 1/1077 : by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids
• CPC Class Codes: C07K 14/64 : Relaxins

Abstract

The present disclosure generally relates to modified relaxin polypeptides, such as modified human relaxin 2 polypeptides, comprising a non-naturally encoded amino acid which is linked to a pharmacokinetic enhancer, and therapeutic uses of such polypeptides, such as for the treatment of cardiovascular conditions (such as heart failure) and/or conditions relating to fibrosis.

Claims

1. A modified relaxin polypeptide comprising a non-naturally encoded amino acid, wherein:

(a) the modified relaxin polypeptide comprises the relaxin A chain polypeptide of SEQ ID NO;

4 and the relaxin B chain polypeptide of SEQ ID NO;

5 or SEQ ID NO;

6, substituted with a non-naturally encoded amino acid at A chain residue 1, and having zero, one, or two additional amino acid changes in said relaxin A chain and having zero, one, or two additional amino acid changes in said relaxin B chain, wherein each of said amino acid changes is independently a substitution, insertion or deletion;

(b) said non-naturally encoded amino acid has the structure;

2. The modified relaxin polypeptide of claim 1, wherein said non-naturally encoded amino acid comprises a carbonyl group, an aminooxy group, a hydrazide group, a hydrazine group, a semicarbazide group, an azide group, or an alkyne group.

3. The modified relaxin polypeptide of claim 1, wherein said non-naturally encoded amino acid comprises a phenylalanine derivative.

4. The modified relaxin polypeptide of claim 1, wherein said non-naturally encoded amino acid is directly linked to said peptide component.

5. The modified relaxin polypeptide of claim 1, wherein said non-naturally encoded amino acid is linked to said pharmacokinetic enhancer through an oxime linkage or triazole linkage.

6. The modified relaxin polypeptide of claim 1, wherein said relaxin A chain polypeptide comprises SEQ ID NO:

35, and said relaxin B chain polypeptide comprises SEQ ID NO;

5 or SEQ ID NO;

6.

7. The modified relaxin polypeptide of claim 1, wherein said peptide component comprises Glu, GGGGS-Glu^γ

(SEQ ID NO;

     139), DRDDRD (SEQ ID NO;

     102), KKKKKK-Glu^γ

(SEQ ID NO;

     103), GGGEEE-Glu^γ

(SEQ ID NO;

     105), EEEGGG-Glu^γ

(SEQ ID NO;

     106), KKKGGG-Glu^γ

(SEQ ID NO;

     107), GGGKKK-Glu^γ

(SEQ ID NO;

     109), Sar-Sar-Sar-Sar-Ser-Glu^γ

(SEQ ID NO;

     112), Sar-Sar-Sar-Glu-Glu-Glu^γ

(SEQ ID NO;

     113), KSGGSGG-Glu^γ

(SEQ ID NO;

     117), dKdKdKdKdKdK-Glu^γ

(SEQ ID NO;

     120), EEEGGG-dGlu^γ

(SEQ ID NO;

     128), EGGGGSK-Glu^γ

(SEQ ID NO;

     130), EEEEEE-Glu^γ

(SEQ ID NO;

     131), KK-Glu^γ

(SEQ ID NO;

     140), KGPKGP-Glu^γ

(SEQ ID NO;

     146), SGGGS-Glu^γ

(SEQ ID NO;

     147), KGGGS-Glu^γ

(SEQ ID NO;

     148), KGGGSE-Glu^γ

(SEQ ID NO;

     149), GSPGSP-Glu^γ

(SEQ ID NO;

     150), GGGGP-Glu^γ

(SEQ ID NO;

     151), EGGS-Glu^γ

(SEQ ID NO;

     152), EGGGP-Glu^γ

(SEQ ID NO;

     153), KGPGSE-Glu^γ

(SEQ ID NO;

     154), Spermine-Glu^γ

, or KKGGS-Glu^γ

(SEQ ID NO;

     156).

8. The modified relaxin polypeptide of claim 1, wherein the relaxin polypeptide comprises the relaxin A chain polypeptide of SEQ ID NO:

4 and the relaxin B chain polypeptide of SEQ ID NO;

5 or SEQ ID NO;

6, substituted with a non-naturally encoded amino acid at A chain residue 1, wherein said non-naturally encoded amino acid is linked to said pharmacokinetic enhancer and said pharmacokinetic enhancer comprises the peptide component GGGGS-Glu^γ

(SEQ ID NO;

     139).

9. The modified relaxin polypeptide of claim 1, wherein said half-life extending moiety comprises a fatty acid or derivative thereof, wherein the fatty acid or derivative thereof is covalently linked to the peptide component.

10. The modified relaxin polypeptide of claim 1, wherein said half-life extending moiety comprises —

C14-COOH.

11. The modified relaxin polypeptide of claim 1, wherein said half-life extending moiety comprises a fatty acid of Formula I:

—

Cn—

COOH 

 

(Formula I)

wherein n is between 12 and 14.

12. The modified relaxin polypeptide of claim 1, wherein said pharmacokinetic enhancer comprises:

13. The modified relaxin polypeptide of claim 1, wherein said half-life extending moiety is conjugated to said peptide component through an amide bond.

14. The modified relaxin polypeptide of claim 1, wherein the relaxin polypeptide comprises the relaxin A chain polypeptide of SEQ ID NO:

4 and the relaxin B chain polypeptide of SEQ ID NO;

5 or SEQ ID NO;

6, substituted with a non-naturally encoded amino acid at A chain residue 1;

wherein said non-naturally encoded amino acid comprises para-acetyl-L-phenylalanine linked to said pharmacokinetic enhancer which comprises

a peptide component comprising GGGGS-Glu^γ

(SEQ ID NO;

     139); and

a half-life extending moiety comprising —

C14-COOH.

15. The modified relaxin polypeptide of claim 1, wherein said modified relaxin polypeptide exhibits an increased in vivo half-life by at least 2-fold compared to the wild-type relaxin polypeptide consisting of the relaxin A chain of SEQ ID NO:

4 linked to the relaxin B chain of SEQ ID NO;

5.

16. A pharmaceutical composition comprising the modified relaxin polypeptide of claim 1 and a pharmaceutically acceptable carrier.

17. A method of manufacturing a modified relaxin polypeptide according to claim 1, comprising:

(a) providing a polypeptide comprising said relaxin A chain and said relaxin B chain, wherein said polypeptide comprises said non-naturally encoded amino acid; and

(b) linking said non-naturally encoded amino acid to said pharmacokinetic enhancer.

18. The modified relaxin polypeptide of claim 1, wherein said modified relaxin polypeptide exhibits an increased in vivo half-life by at least 5-fold compared to the wild-type relaxin polypeptide consisting of the relaxin A chain of SEQ ID NO:

4 linked to the relaxin B chain of SEQ ID NO;

5.

19. The modified relaxin polypeptide of claim 1, wherein said modified relaxin polypeptide exhibits an increased in vivo half-life by at least 10-fold compared to the wild-type relaxin polypeptide consisting of the relaxin A chain of SEQ ID NO:

4 linked to the relaxin B chain of SEQ ID NO;

5.

20. A modified relaxin polypeptide comprising:

21. The modified relaxin polypeptide of claim 20, wherein AQ1-Relaxin comprises a relaxin A chain polypeptide of SEQ ID NO:

35 and a relaxin B chain polypeptide of SEQ ID NO;

5 or SEQ ID NO;

6.

22. A pharmaceutical composition comprising the modified relaxin polypeptide of claim 20 and a pharmaceutically acceptable carrier.

23. A modified relaxin polypeptide comprising AQ1-GGGGS-Glu^γ

(SEQ ID NO;

     139)-C14-COOH, which comprises the structure shown in FIG. 9, or which comprises a relaxin A chain polypeptide of SEQ ID NO;

35 and a relaxin B chain polypeptide of SEQ ID NO;

6, wherein the modified para-acetyl-phenylalanine located at the N-terminus of said relaxin A chain polypeptide is linked to the pharmacokinetic enhancer depicted in Formula III;

24. The modified relaxin polypeptide of claim 23, which comprises the structure shown in FIG. 9.

25. The modified relaxin polypeptide of claim 24, which consists of the structure shown in FIG. 9.

26. The modified relaxin polypeptide of claim 25, wherein said modified relaxin polypeptide exhibits an increased in vivo half-life by at least 2-fold compared to the wild-type relaxin polypeptide consisting of the relaxin A chain of SEQ ID NO:

4 linked to the relaxin B chain of SEQ ID NO;

5.

27. A pharmaceutical composition comprising the modified relaxin polypeptide of claim 25 and a pharmaceutically acceptable carrier.

28. The modified relaxin polypeptide of claim 24, wherein said modified relaxin polypeptide exhibits an increased in vivo half-life by at least 2-fold compared to the wild-type relaxin polypeptide consisting of the relaxin A chain of SEQ ID NO:

4 linked to the relaxin B chain of SEQ ID NO;

5.

29. A pharmaceutical composition comprising the modified relaxin polypeptide of claim 24 and a pharmaceutically acceptable carrier.

30. A pharmaceutical composition comprising the modified relaxin polypeptide of claim 23 and a pharmaceutically acceptable carrier.