Process for preparing human relaxin-2

摘要

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-CysB 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; andsubsequent purification of the relaxin-2 formed.

主权项

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.