A kind of method for solid-phase synthesis of ziconotide by fragment method

Abstract

The invention belongs to the field of polypeptide synthesis, and relates to a solid-phase synthesis method of ziconotide by a segment process. The method aims to solve the technical problems of long synthesis period and low crude peptide purity in the existing preparation method, and the technical problems of complex operation, low total yield and the like in the disulfide-bond formation process. The technical scheme is as follows: the method comprises the following steps: synthesizing four segment peptides [19-25]A, [12-18]B, [6-11]C and [1-5]D of ziconotide by a solid-phase process; sequentially connecting the all-protected peptide fragments B, C and D to the peptide resin A, and cracking to obtain a ziconotide linear peptide; and finally, carrying out liquid-phase one-step oxidization, which has the advantages of mild conditions, simplified operation, easy after-treatment and environment friendliness, to obtain the ziconotide. The technical scheme can greatly shorten the synthesis period, lower the synthesis and purification difficulty and the production cost, and enhance the purity of the linear peptide and the total yield of the product. The method is beneficial to industrial large-scale production.

Description

technical field [0001] The invention relates to the field of polypeptide synthesis, in particular to a method for solid-phase synthesis of ziconotide by a fragment method. Background technique [0002] Ziconotide (Ziconotide), trade name Prialt, was developed by Elan Company of Ireland. It was first approved for marketing in the United States on January 11, 2005. It was approved by the European Union in February 2005. In 2007, the special group of the Joint Analgesia Conference recommended Qi Conotide as a first-line intrathecal analgesic. It is an N-type calcium channel blocker, a synthetic form of the omega-conotoxin MVIIA isolated from the conus snail mollusk, and can be used as a non-opioid analgesic drug for intrathecal administration. Ziconotide specifically and effectively binds to N-VSCC (N-type voltage-sensitive calcium channels) to block the release of pain-causing transmitters, and then realizes its pharmacological effects. It is suitable for intrathecal injectio...

AI Technical Summary

Problems solved by technology

[0006] Regarding the method of oxidation to form disulfide bonds in the synthesis step of ziconotide, patents CN200710301598.8, CN201110139661.9 and CN201310202216.1 all disclose the use of Cys with three different sulfhydryl protecting groups to synthesize linear peptides, and utilize the self-stabilization of the protecting groups properties, the preparation strategy of stepwise oxidation to form three pairs of disulfide bonds is adopted; although the disulfide bonds formed have good orientation, this strategy has many intermediate steps and is cumbersome to operate

Patent CN200910188686.0 adopts the preparation strategy of one-step oxidation to form three pairs of disulfide bonds at the same time. Although the process is simplified, the metal salt compounds used to remove Acm are very harmful to the human body and are not conducive to environmental protection.

Patent CN201310202089.5 discloses iodine, H 2 o 2 etc. as an oxidant, oxidize to form a disulfide bond, but this type of oxidant can easily cause the oxidation of Met in the ziconotide peptide chain

[0007] The above technical scheme has the following disadvantages: in the process of synthesizing the peptide chain, the method of coupling amino acids one by one is adopted, the synthesis cycle is long, and the purity of the linear crude peptide is low; there are complicated operations in the process of disulfide bond formation, and it is difficult to realize large-scale production. The technical problem of low total yield

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