Preparation method for bivalirudin

Abstract

The invention belongs to the technical field of polypeptide medicament preparation methods, and in particular relates to a preparation method for bivalirudin. The preparation method for the bivalirudin comprises solid phase polypeptide synthesis for preparing bivalirudin resin, acid hydrolysis of the bivalirudin resin to obtain a crude bivalirudin product, and purification of the crude bivalirudin product to obtain a purified bivalirudin product, wherein the solid phase polypeptide synthesis for preparing the bivalirudin resin comprises the following steps of: sequentially connecting corresponding Fmoc- protected amino acids in the following sequences to Fmoc-Leu-carrier resin by a solid phase coupling synthesis method: R1-D-Phe-Pro-Arg(Pbf)-Pro-X-Asn(R2)-Gly-Asp(OtBu)-, Phe-Glu(OtBu)-Glu(OtBu)-Ile-Pro-Glu(OtBu)-Glu(OtBu)- and Tyr(tBu)-Leu-resin, and thus obtaining the bivalirudin resin; and when the X fragment is connected, only one times of solid phase coupling synthesis reaction is used, and the corresponding Fmoc- protected amino acid is Fmoc-Gly-Gly-Gly-Gly-OH. The purity of the bivalirudin is more than 99.5 percent, and the single impurity is less than 0.2 percent.

Description

technical field [0001] The invention belongs to the technical field of polypeptide drug preparation methods, in particular to a preparation method of bivalirudin. Background technique [0002] Bivalirudin is a direct thrombin inhibitor (DTI), consisting of 20 amino acid residues. Direct thrombin inhibitors (DTI) are a group of polypeptides that directly inhibit thrombin activity without the participation of cofactors. They not only have anticoagulant function, but also inhibit platelet aggregation. Compared with other anticoagulant drugs, its pharmacological action and pharmacokinetic characteristics are superior, especially in cardiovascular diseases. [0003] Bivalirudin is a hirudin derivative (fragment), an artificially synthesized polypeptide containing 20 amino acid residues. It is mainly composed of two parts, one part is a short peptide chain at the amino terminal (N-terminal), containing a phenylalanine-proline-arginine-proline sequence, which can specifically bin...

AI Technical Summary

Problems solved by technology

[0013] The structure of bivalirudin contains a Gly-Gly-Gly-Gly fragment. In the process of sequentially inserting Fmoc-Gly in the solid phase method, due to the characteristics of Gly itself, the following impurities are produced in the product: [+1Gly]-bivalirudin Ludin, [+2Gly]-bivalirudin, [-1Gly]-bivalirudin, [-2Gly]-bivalirudin, and these impurities are similar to the polarity of bivalirudin itself, so in the purification When it is difficult to completely purify it, the total yield of the product cannot be effectively improved, and the purity of the product is reduced, which affects the safety of the drug

[0014] None of the above patents can solve the problem of increase and decrease of Gly in the structure of bivalirudin. In order to solve the above problems, Israel NOVETIDE Company WO2010117725 patent uses Fmoc-Gly-Gly-OH as raw material to insert Gly-Gly- Gly-Gly fragments, but only resolve [+1Gly]-bivalirudin and [-1Gly]-bivalirudin impurities, but not [+2Gly]-bivalirudin and [-2Gly]- Production of bivalirudin

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