Method for preparing bivalirudin

Abstract

The invention belongs to the technical field of preparation methods for polypeptide medicines, and particularly relates to a method for preparing bivalirudin. The method for preparing the bivalirudin comprises the following steps of: preparing a bivalirudin resin by a solid phase polypeptide synthesis method, performing acidolysis on the bivalirudin resin to obtain a bivalirudin crude product, and purifying the bivalirudin crude product to obtain a bivalirudin pure product, wherein the step of preparing the bivalirudin resin by the solid phase polypeptide synthesis method comprises the following substeps of: sequentially accessing the corresponding protection amino acid or fragments on an Fmoc-Leu-carrier resin by a solid phase coupling and synthesis method to obtain the bivalirudin resin, wherein the corresponding protection amino acid or fragments have the following sequences: R1-D-Phe-Pro-X-Y-Phe-Glu(OtBu)-Glu(OtBu)-Ile-Pro-Glu(OtBu)-Glu(OtBu)-Tyr(tBu)-Leu-resin, R1 is Fmoc, Boc or H; X is Arg(Pbf)-Pro; Y is Gly-Gly-Gly-Asn(R2)-Gly-Asp-(OtBu); and R2 is Trt or H. The purity of the product can reach above 99.5 percent.

Description

technical field [0001] The invention belongs to the technical field of polypeptide drug preparation methods, and in particular relates to a new 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 ...

AI Technical Summary

Problems solved by technology

[0009] Due to the characteristics of the structure of this product, this variety contains 5 Gly. During the process of sequentially adding Fmoc-Gly by the solid-phase method, due to the characteristics of Gly itself, the following impurities are produced in the product: [+1Gly]-bivalirudin, [+2Gly]-bivalirudin, [-1Gly]-bivalirudin, [-2Gly]-bivalirudin, and these impurities are similar in polarity to bivalirudin itself, so it is very difficult to purify It is difficult to completely purify, the total yield of the product cannot be effectively improved, and the product purity is not high, which affects the safety of medication

[0010] In order to solve the problem of increase and decrease of Gly in the structure of bivalirudin, NOVETIDE company WO2010117725 patent of Israel, using Fmoc-Gly-Gly-OH as the raw material in the peptide grafting process only solves the problem of [+1Gly]-bivalirudin and Production of [-1Gly]-bivalirudin impurity without resolution of [+2Gly]-bivalirudin and [-2Gly]-bivalirudin production

[0011] At the same time, it was also found in the preparation process that due to the large steric hindrance of amino acids, the reaction was not complete during the insertion of Fmoc-Arg(Pbf)-OH, and [-Arg]-bivalirudin impurities were easily produced, which seriously affected the product. yield