# US 12,097,242 B2 · Treatment of fibrosis, cardiovascular disease and heart failure with modified relaxin polypeptides

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- Number: US 12,097,242 B2
- Document ID: US12097242B2
- Status: Granted
- Kind: B2
- Application number: 17/721,569
- Filed: 2022-04-15
- Published: 2022-10-20
- Granted: 2024-09-24
- Earliest claimed: 2017-02-08
- Assignees: Bristol-Myers Squibb Company
- Applicants: Bristol-Myers Squibb Company
- Inventors: Gene M. Dubowchik; Olafur S. Gudmundsson; Xiaojun Han; R. Michael Lawrence; Dasa Lipovsek; Cort S. Madsen; Claudio Mapelli; Paul E. Morin; Michael C. Myers
- Web page: https://patentgrep.com/patent/US12097242B2

## CPC

- A61K 38/18: Growth factors; Growth regulators
- A61K 38/2221: {Relaxins}
- A61P 1/04: for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
- A61P 1/16: for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
- A61P 1/18: for pancreatic disorders, e.g. pancreatic enzymes
- A61P 3/06: Antihyperlipidemics
- A61P 9/04: Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
- A61P 9/06: Antiarrhythmics
- A61P 9/10: for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
- A61P 9/12: Antihypertensives
- A61P 11/00: Drugs for disorders of the respiratory system
- A61P 13/12: of the kidneys
- A61P 17/00: Drugs for dermatological disorders
- A61P 17/02: for treating wounds, ulcers, burns, scars, keloids, or the like
- A61P 19/02: for joint disorders, e.g. arthritis, arthrosis
- A61P 25/02: for peripheral neuropathies
- A61P 27/02: Ophthalmic agents
- A61P 27/12: for cataracts
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C07K 1/1077: {by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids}
- C07K 14/64: Relaxins
- 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)
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- 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)
- A61P 9/00: Drugs for disorders of the cardiovascular system
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)

## 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

### Claim 1

A method of treating fibrosis, comprising administering an effective amount of a biologically active 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:
wherein the R group is any substituent other than the side chain found in alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine or valine; and
(c) said non-naturally encoded amino acid is linked to a pharmacokinetic enhancer comprising a peptide component between 4 and 8 amino acids and a half-life extending moiety, comprising a fatty acid of Formula I:
-Cn-COOH (Formula I)
wherein n is between 12 and 16,
said peptide component comprises Gluγ, and
wherein said modified relaxin polypeptide is administered to a patient suffering from fibrosis.

### Claim 2 (depends on 1)

The method of claim 1, wherein said non-naturally encoded amino acid comprises a phenylalanine derivative.

### Claim 3 (depends on 1)

The method 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.

### Claim 4 (depends on 1)

The method of claim 1, wherein said peptide component comprises GGGGS-Gluγ (SEQ ID NO: 139), DRDDRD (SEQ ID NO: 102), KKKKKK-Gluγ (SEQ ID NO: 103), GGGEEE-Gluγ (SEQ ID NO: 105), EEEGGG-Gluγ (SEQ ID NO: 106), KKKGGG-Gluγ (SEQ ID NO: 107), GGGKKK-Gluγ (SEQ ID NO: 109), Sar-Sar-Sar-Sar-Ser-Gluγ (SEQ ID NO: 112), Sar-Sar-Sar-Glu-Glu-Gluγ (SEQ ID NO: 113), KSGGSGG-Gluγ (SEQ ID NO: 117), dKdKdKdKdKdK-Gluγ (SEQ ID NO: 120), EEEGGG-d Gluγ (SEQ ID NO: 128), EGGGGSK-Gluγ (SEQ ID NO: 130), EEEEEE-Gluγ (SEQ ID NO: 131), KK-Gluγ (SEQ ID NO: 140), KGPKGP-Gluγ (SEQ ID NO: 146), SGGGS-Gluγ (SEQ ID NO: 147), KGGGS-Gluγ (SEQ ID NO: 148), KGGGSE-Gluγ (SEQ ID NO: 149), GSPGSP-Gluγ (SEQ ID NO: 150), GGGGP-Gluγ (SEQ ID NO: 151), EGGS-Gluγ (SEQ ID NO: 152), EGGGP-Gluγ (SEQ ID NO: 153), KGPGSE-Gluγ (SEQ ID NO: 154), Spermine-Gluγ, or KKGGS-Gluγ (SEQ ID NO: 156).

### Claim 5 (depends on 1)

The method of claim 1, wherein 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, wherein said non-naturally encoded amino acid is linked to said pharmacokinetic enhancer and said pharmacokinetic enhancer comprises the peptide component GGGGS-Gluγ (SEQ ID NO: 139).

### Claim 6 (depends on 1)

The method of claim 1, wherein said pharmacokinetic enhancer comprises:
wherein the aminooxy group is linked to the non-natural amino acid in said modified relaxin polypeptide.

### Claim 7 (depends on 1)

The method of claim 1, wherein 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;
said non-naturally encoded amino acid comprises para-acetyl-L-phenylalanine and is linked to said pharmacokinetic enhancer which comprises
a peptide component comprising GGGGS-Gluγ (SEQ ID NO: 139); and
a half-life extending moiety comprising —C14-COOH.

### Claim 8 (depends on 1)

The method of claim 1, wherein the modified 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, wherein a para-acetyl-L-phenylalanine is located at position 1 of the relaxin A chain polypeptide, which para-acetyl-L-phenylalanine is linked to the pharmacokinetic enhancer GGGGS-Gluγ (SEQ ID NO: 139)-C14-COOH, as depicted in Formula III:

### Claim 9

A method of treating a cardiovascular disease comprising administering an effective amount of a biologically active 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:
wherein the R group is any substituent other than the side chain found in alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine or valine; and
(c) said non-naturally encoded amino acid is linked to a pharmacokinetic enhancer comprising a peptide component between 4 and 8 amino acids and a half-life extending moiety, comprising a fatty acid of Formula I:
-Cn-COOH (Formula I)
wherein n is between 12 and 16,
said peptide component comprises Gluγ, and
further wherein said modified relaxin polypeptide is administered to a patient suffering from a cardiovascular disease.

### Claim 10 (depends on 9)

The method of claim 9, wherein said cardiovascular disease is selected from coronary artery disease, heart attack, arrhythmia, heart failure, cardio-renal syndrome, post myocardial remodeling, and cardiomyopathy.

### Claim 11 (depends on 9)

The method of claim 9, wherein said non-naturally encoded amino acid comprises a phenylalanine derivative.

### Claim 12 (depends on 9)

The method of claim 9, 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.

### Claim 13 (depends on 9)

The method of claim 9, wherein said peptide component comprises GGGGS-Gluγ (SEQ ID NO: 139), DRDDRD (SEQ ID NO: 102), KKKKKK-Gluγ (SEQ ID NO: 103), GGGEEE-Gluγ (SEQ ID NO: 105), EEEGGG-Gluγ (SEQ ID NO: 106), KKKGGG-Gluγ (SEQ ID NO: 107), GGGKKK-Gluγ (SEQ ID NO: 109), Sar-Sar-Sar-Sar-Ser-Gluγ (SEQ ID NO: 112), Sar-Sar-Sar-Glu-Glu-Gluγ (SEQ ID NO: 113), KSGGSGG-Gluγ (SEQ ID NO: 117), dKdKdKdKdKdK-Gluγ (SEQ ID NO: 120), EEEGGG-d Gluγ (SEQ ID NO: 128), EGGGGSK-Gluγ (SEQ ID NO: 130), EEEEEE-Gluγ (SEQ ID NO: 131), KK-Gluγ (SEQ ID NO: 140), KGPKGP-Gluγ (SEQ ID NO: 146), SGGGS-Gluγ (SEQ ID NO: 147), KGGGS-Gluγ (SEQ ID NO: 148), KGGGSE-Gluγ (SEQ ID NO: 149), GSPGSP-Gluγ (SEQ ID NO: 150), GGGGP-Gluγ (SEQ ID NO: 151), EGGS-Gluγ (SEQ ID NO: 152), EGGGP-Gluγ (SEQ ID NO: 153), KGPGSE-Gluγ (SEQ ID NO: 154), Spermine-Gluγ, or KKGGS-Gluγ (SEQ ID NO: 156).

### Claim 14 (depends on 9)

The method of claim 9, wherein 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, wherein said non-naturally encoded amino acid is linked to said pharmacokinetic enhancer and said pharmacokinetic enhancer comprises the peptide component GGGGS-Gluγ (SEQ ID NO: 139).

### Claim 15 (depends on 9)

The method of claim 9, wherein said pharmacokinetic enhancer comprises:
wherein the aminooxy group is linked to the non-natural amino acid in said modified relaxin polypeptide.

### Claim 16 (depends on 9)

The method of claim 9, wherein 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;
said non-naturally encoded amino acid comprises para-acetyl-L-phenylalanine and is linked to said pharmacokinetic enhancer which comprises
a peptide component comprising GGGGS-Gluγ (SEQ ID NO: 139); and
a half-life extending moiety comprising —C14-COOH.

### Claim 17 (depends on 9)

The method of claim 9, wherein the modified 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, wherein a para-acetyl-L-phenylalanine is located at position 1 of the relaxin A chain polypeptide, which para-acetyl-L-phenylalanine is linked to the pharmacokinetic enhancer GGGGS-Gluγ (SEQ ID NO: 139)-C14-COOH, as depicted in Formula III:

### Claim 18

A method of treating heart failure comprising administering an effective amount of a biologically active 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 land 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:
wherein the R group is any substituent other than the side chain found in alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine or valine; and
(c) said non-naturally encoded amino acid is linked to a pharmacokinetic enhancer comprising a peptide component between 4 and 8 amino acids and a half-life extending moiety, comprising a fatty acid of Formula I:
-Cn-COOH (Formula I)
wherein n is between 12 and 16,
said peptide component comprises Gluγ, and
further wherein said modified relaxin polypeptide is administered to a patient suffering from heart failure.

### Claim 19 (depends on 18)

The method of claim 18, wherein said heart failure is selected from advanced heart failure, heart failure with impaired kidney function, chronic heart failure, chronic heart failure with mid-range ejection fraction (HFmEF), acute heart failure, post-acute heart failure, compensated heart failure, decompensated heart failure, right heart failure, left heart failure, global heart failure, heart failure associated with congenital heart defects, heart failure associated with heart valve defects, heart failure associated with combined heart valve defects, diabetic heart failure, heart failure associated with cardiac storage disorders, diastolic heart failure, systolic heart failure, heart failure with preserved ejection fraction (HFpEF), and heart failure with reduced ejection fraction (HFrEF).

### Claim 20 (depends on 18)

The method of claim 18, wherein said non-naturally encoded amino acid comprises a phenylalanine derivative.

### Claim 21 (depends on 18)

The method of claim 18, 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.

### Claim 22 (depends on 18)

The method of claim 18, wherein said peptide component comprises GGGGS-Gluγ (SEQ ID NO: 139), DRDDRD (SEQ ID NO: 102), KKKKKK-Gluγ (SEQ ID NO: 103), GGGEEE-Gluγ (SEQ ID NO: 105), EEEGGG-Gluγ (SEQ ID NO: 106), KKKGGG-Gluγ (SEQ ID NO: 107), GGGKKK-Gluγ (SEQ ID NO: 109), Sar-Sar-Sar-Sar-Ser-Gluγ (SEQ ID NO: 112), Sar-Sar-Sar-Glu-Glu-Gluγ (SEQ ID NO: 113), KSGGSGG-Gluγ (SEQ ID NO: 117), dKdKdKdKdKdK-Gluγ (SEQ ID NO: 120), EEEGGG-d Gluγ (SEQ ID NO: 128), EGGGGSK-Gluγ (SEQ ID NO: 130), EEEEEE-Gluγ (SEQ ID NO: 131), KK-Gluγ (SEQ ID NO: 140), KGPKGP-Gluγ (SEQ ID NO: 146), SGGGS-Gluγ (SEQ ID NO: 147), KGGGS-Gluγ (SEQ ID NO: 148), KGGGSE-Gluγ (SEQ ID NO: 149), GSPGSP-Gluγ (SEQ ID NO: 150), GGGGP-Gluγ (SEQ ID NO: 151), EGGS-Gluγ (SEQ ID NO: 152), EGGGP-Gluγ (SEQ ID NO: 153), KGPGSE-Gluγ (SEQ ID NO: 154), Spermine-Gluγ, or KKGGS-Gluγ (SEQ ID NO: 156).

### Claim 23 (depends on 18)

The method of claim 18, wherein 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, wherein said non-naturally encoded amino acid is linked to said pharmacokinetic enhancer and said pharmacokinetic enhancer comprises the peptide component GGGGS-Gluγ (SEQ ID NO: 139).

### Claim 24 (depends on 18)

The method of claim 18, wherein said pharmacokinetic enhancer comprises:
wherein the aminooxy group is linked to the non-natural amino acid in said modified relaxin polypeptide.

### Claim 25 (depends on 18)

The method of claim 18, wherein the modified 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;
said non-naturally encoded amino acid comprises para-acetyl-L-phenylalanine and is linked to said pharmacokinetic enhancer which comprises
a peptide component comprising GGGGS-Gluγ (SEQ ID NO: 139); and
a half-life extending moiety comprising —C14-COOH.

### Claim 26 (depends on 18)

The method of claim 18, wherein the modified 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, wherein a para-acetyl-L-phenylalanine is located at position 1 of the relaxin A chain polypeptide, which para-acetyl-L-phenylalanine is linked to the pharmacokinetic enhancer GGGGS-Gluγ (SEQ ID NO: 139)-C14-COOH, as depicted in Formula III: