# US 9,434,780 B2 · Process for preparing human relaxin-2

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- Number: US 9,434,780 B2
- Document ID: US9434780B2
- Status: Granted
- Kind: B2
- Application number: 14/234,919
- Filed: 2012-08-03
- Published: 2014-11-06
- Granted: 2016-09-06
- Earliest claimed: 2011-08-04
- Assignees: Relaxera Pharma GmbH & Co. KG i.G.
- Applicants: Wolf-Georg Forssmann; Thomas Dschietzig; Ludger Ständker; Andreas Zgraja; Jochen Hirsch
- Inventors: Wolf-Georg Forssmann; Thomas Dschietzig; Ludger Ständker; Andreas Zgraja; Jochen Hirsch
- Web page: https://patentgrep.com/patent/US9434780B2

## CPC

- C07K 14/64: Relaxins
- A61P 1/16: for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
- A61P 3/10: for hyperglycaemia, e.g. antidiabetics
- A61P 9/00: Drugs for disorders of the cardiovascular system
- A61P 9/04: Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
- 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 11/10: Expectorants
- A61P 13/12: of the kidneys
- Y02P 20/55: Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
- 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)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- Y02P: CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS

## Abstract

A process for preparing human relaxin-2 having the following amino acid sequence: A chain: (SEQ ID NO: 1) pGlu-Leu-Tyr-Ser-Ala-Leu-Ala-Asn-Lys-Cys-Cys-His- Val-Gly-Cys-Thr-Lys-Arg-Ser-Leu-Ala-Arg-Phe-Cys B chain: (SEQ ID NO: 2) Asp-Ser-Trp-Met-Glu-Glu-Val-Ile-Lys-Leu-Cys-Gly- Arg-Glu-Leu-Val-Arg-Ala-Gln-Ile-Ala-Ile-Cys-Gly- Met-Ser-Thr-Trp-Ser; comprising the following steps: providing the amino acids necessary for the synthesis of the A and B chains with usual protective groups, wherein the cysteines are employed as trityl-protected amino acids (L-Cys(Trt)-OH); effecting a chromatographic purification of the individual chains A and B after the solid state synthesis; followed by the simultaneous folding and combination of the individual chains A and B in ammonium hydrogencarbonate buffer at pH 7.9 to 8.4; and subsequent purification of the relaxin-2 formed.

## Claims

### Claim 1

A method of treating a disease comprising synthesizing Relaxin-2 and administering the Relaxin-2 for the treatment of pulmonary hypertension, rental fibrosis, or acute heart failure,
with the relaxin-2 having the following amino acid sequence: an A chain having SEQ ID NO:1 wherein the first residue Glu is a pyroglutamic acid and a B chain having SEQ ID NO:2, with the synthesizing comprising the following steps:
providing the amino acids necessary for the synthesis of the A and B chains with protective groups, wherein the cysteines are employed as trityl-protected amino acids (L-Cys(Trt)-OH); effecting a solid state synthesis of the individual chains A and B;
effecting a chromatographic purification of the individual chains A and B after the solid state synthesis;
followed by the simultaneous folding and combination of the individual chains A and B in ammonium hydrogencarbonate buffer at pH 7.9 to 8.4;
addition of cystine and cysteine as a redox pair at a concentration of 2 mg cystine and 2 mg cysteine per mg of peptide employed (A+B chains) and
subsequent purification of the relaxin-2 formed.

### Claim 2 (depends on 1)

The method of claim 1 with the Relaxin-2 being in a lyophilized form with an excipient.

### Claim 3 (depends on 2)

The method of claim 2 with the Relaxin-2 being reconstituted in aqueous solution.

### Claim 4 (depends on 1)

The method of claim 1 with the Relaxin-2 being encapsulated in liposomes.

### Claim 5 (depends on 4)

The method of claim 4 with the Relaxin-2 being in an aqueous solution containing relaxin-2 encapsulated in liposomes, or in an ointment containing relaxin-2 encapsulated in liposomes.