Method for preparing carbetocin

Abstract

The invention relates to a solid-phase synthesis method of carbetocin, which comprises the following steps: reacting a PAM (polyacrylamide) resin and Fmoc-Gly-OH to obtain an Fmoc-Gly-PAM resin; sequentially connecting amino acid with Fmoc protective group by solid-phase synthesis to obtain a BrCH2CH2CH2CONH-Tyr(OMe)-Ile-Gln-Asn-Cys(Mmt)-Pro-Leu-Gly-PAM resin; removing the Mmt protective group in the Cys; carrying out cyclization reaction by using DMAP (dimethylaminopyridine) as a cyclization reagent to obtain Cyclo[CH2CH2CH2CO-Tyr(OMe)-Ile-Gln-Asn-Cys]-Pro-Leu-Gly-PAM; carrying out ammonolysis on the cyclization product with an ammonia methanol solution to obtain a carbetocin crude product; and purifying and freeze-drying to obtain the carbetocin. The total yield is up to higher than 45%. Compared with the conventional preparation method, the method provided by the invention avoids using abundant high-concentration strongly-corrosive acid and flammable and explosive aether, has the advantages of high yield, low cost, mild reaction conditions and small environmental pollution, and is beneficial to industrialized production.

Description

technical field [0001] The invention relates to a method for solid-phase synthesis of polypeptides, in particular to a method for synthesizing carbetocin. Background technique [0002] Carbetocin (Carbetocin) was developed by Swedish Ferring Pharmaceutical Co., Ltd. and was used clinically in 1990. It is a synthetic long-acting oxytocin nonapeptide analogue with agonist properties, used for selective hard After cesarean section under extramembranous or spinal anesthesia, it is used to prevent uterine atony and postpartum hemorrhage, with quick onset, long duration of action, and safe and effective clinical application. [0003] The molecular formula of carbetocin is C45H69N11O12S, the molecular weight is 998.17, the chemical name is: desammonium-2-oxo-methyltyrosine-1-к oxytocin, and the structure is: [0004] [0005] Patent No. ZL200910106889.0 discloses a method for preparing carbetocin in solid phase, using amino resin as the starting material, connecting amino acids...

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Problems solved by technology

[0005] Patent No. ZL200910106889.0 discloses a method for preparing carbetocin in solid phase, using amino resin as the starting material, connecting amino acids with Fmoc protecting groups in sequence, removing half of the carbetocin with tetrakis(triphenylphosphine) palladium Cystine side chain allyl protecting group, cyclization, cleavage, purification and lyophilization to obtain refined carbetocin, the reagent for removing the side chain allyl protecting group in this method is expensive, and the cleavage reagent used is The mixed solution of trifluoroacetic acid, water, phenol, ethanedithiol and sulfide anisole is a high-concentration strong corrosive acid, which is easy to burn the human body, has high requirements on the equipment used, causes great environmental pollution, increases costs, and is not conducive to large-scale production.

[0006] The preparation method of carbetocin disclosed in the application number 201110151928.6 adopts a novel sulfhydryl-protected amino acid Fmoc-Cys ((CH2)3COOAll), and the price of the side chain allyl-protecting group reagent in this method is also very expensive, which is not conducive to large-scale mass production

[0007] Application No. 201110001400.0 discloses a preparation method for synthesizing carbetocin, which uses a solid phase to form a linear carbetocin precursor peptide, and liquid phase cyclization, which requires a very dilute solvent (10 -4 ~10 -5 mol / L), a large amount of waste liquid is produced, and polymers are easily produced at the same time, resulting in low cyclization efficiency and complicated post-treatment.

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