Method for solid phase synthesis of plecanatide by means of secondary cyclization

A technology of solid-phase synthesis and plecanatide, which is applied in peptide preparation methods, chemical instruments and methods, peptides, etc., can solve problems such as low efficiency, cumbersome extraction process, and difficulty in obtaining high-purity products, and achieve directional efficiency High, high product yield, conducive to the effect of product purification

Inactive Publication Date: 2017-11-24
SUZHOU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, plecanatide is obtained through biological extraction technology. The biological extraction is affected by the existing limited biological resources. The extraction process is cumbersome, the efficiency is low, and it is difficult to obtain high-purity products.

Method used

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  • Method for solid phase synthesis of plecanatide by means of secondary cyclization
  • Method for solid phase synthesis of plecanatide by means of secondary cyclization
  • Method for solid phase synthesis of plecanatide by means of secondary cyclization

Examples

Experimental program
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Effect test

Embodiment 1

[0055] Example 1: Synthesis of Fmoc-Leu-king resin with a degree of substitution of 0.10mmol / g

[0056] Weigh 20g of Wang resin with a degree of substitution of 0.45mmol / g, add to the solid phase reaction column, add to the solid phase reaction column, wash once with DMF, swell the resin with DMF for 30 minutes, and take 15.81g Fmoc-Leu -OH (45mmol), 6.01g HOBt (45mmol) dissolved in DMF, add 7.0ml DIC (45mmol) under ice water bath to activate, add to the above reaction column with resin, add 2.75g DMAP (22.5mmol) after 5 minutes After reacting for 2 hours, it was washed with DMF 3 times, DCM washed 3 times, capped with 100ml acetic anhydride / pyridine overnight, and methanol was shrunk and dried to obtain Fmoc-Leu-king resin with a detection degree of substitution of 0.10mmol / g.

Embodiment 2

[0057] Example 2: Synthesis of Fmoc-Leu-king resin with a degree of substitution of 0.50mmol / g

[0058] Weigh 10g of Wang resin with a degree of substitution of 1.50mmol / g, add to the solid phase reaction column, add to the solid phase reaction column, wash once with DMF, swell the resin with DMF for 30 minutes, and take 26.48g Fmoc-Leu -OH (75mmol), 10.13g HOBt (75mmol) dissolved in DMF, add 11.6ml DIC (75mmol) under ice water bath to activate, add to the above reaction column with resin, add 4.5g DMAP (37.5mmol) after 5 minutes After reacting for 2 hours, it was washed 3 times with DMF and 3 times with DCM, capped overnight with 100ml of acetic anhydride / pyridine, and methanol was shrinked and dried to obtain 22.54g of Fmoc-Leu-king resin. The detection degree of substitution was 0.50mmol / g.

Embodiment 3

[0059] Example 3: Preparation of fully protected pukanatide linear peptide king resin

[0060] Weigh 10.00g (1mmol) of Fmoc-Leu-King resin with a degree of substitution of 0.10mmol / g, add it to the solid phase reaction column, wash with DMF once, swell Fmoc-Leu-King resin with DMF for 30 minutes, then use DMF : The mixed solution of 4:1 volume ratio of pyridine removes Fmoc protection, then washes with DMF 6 times, weighs 2.07g Fmoc-Cys(Acm)-OH (5mmol), 0.68g HOBt (5mmol) and adds volume ratio of 1 :1 mixed solution of DCM and DMF, add 0.8ml DIC (5mmol) under ice-water bath for activation, add it to the above reaction column containing resin, react for 2 hours at room temperature, then detect the end of the reaction by ninhydrin method, if If the resin is colorless and transparent, it means that the reaction is complete; if the resin is colored, it means that the reaction is incomplete and requires another 1 hour to react. This criterion is suitable for subsequent amino acid coup...

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Abstract

The invention provides a synthetic method of plecanatide. The method comprises the following sequential steps of: performing a programmed reaction on Wang resin as a solid phase carrier, performing condensation reaction successively to connect 16 protective amino acids to obtain linear peptide fully protected resin with 16 amino acids, wherein in two groups of Cys forming a disulfide bond, Cys in the same group are connected to Mmt or Acm at the same time, and the protecting groups connected by Cys in different groups are different; and successively removing the protecting groups of the two groups of Cys, successively performing directional cyclization reactions to form a dithio cyclic bond, and removing the protecting groups on the side chain and cutting resin to obtain plecanatide containing two dithiorings. The method provided by the invention has the beneficial effects that the method can generate few byproducts, and is high in directional efficiency and simple; the product is favorably purified, and the product yield is high.

Description

Technical field [0001] The invention relates to a method for preparing polypeptide drugs, in particular to a method for synthesizing pukanatide through solid phase secondary cyclization. Background technique [0002] Pulcanatide, English name: plecanatide. [0003] The peptide sequence is: [0004] H-Asn-Asp-Glu-Cys(1)-Glu-Leu-Cys(2)-Val-Asn-Val-Ala-Cys(1)-Thr-Gly-Cys(2)-Leu-OH. [0005] The structural formula is as follows: [0006] [0007] Plecanatide was developed by Synergy Pharmaceuticals in the United States. It is an analog of uroguanylin, a cyclic polypeptide containing 16 amino acids, and has a natriuretic guanylate cyclase receptor activation. The role of the drug can adjust the acid-base ions in the gastrointestinal tract, induce fluid transport into the gastrointestinal tract, and increase the peristalsis of the gastrointestinal tract. It is suitable for the treatment of adult chronic idiopathic constipation. The US Food and Drug Administration (FDA) approved the listing...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K1/04C07K1/06C07K1/08
CPCC07K7/08Y02P20/55
Inventor 陈丰钱君超刘成宝陈志刚
Owner SUZHOU UNIV OF SCI & TECH
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