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A kind of preparation method of adenylyl succinic acid or salt

A technology of adenylate succinic acid and Escherichia coli, which is applied in the field of large-scale production of candidate drugs for treating glucose and lipid metabolism disorders, can solve the problems of difficulty in guaranteeing the source of raw materials, high price, etc., and achieve breakthrough in monopoly sales and low cost , the recovery rate is good

Active Publication Date: 2021-04-02
INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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Problems solved by technology

[0004] In order to solve the problem that the existing adenylyl succinic acid, that is, S-AMP, is difficult to produce, its price is expensive, and it is difficult to ensure the source of raw materials for its druggability research, the present invention provides a preparation method of adenylyl succinic acid or a salt, It is a small-scale synthesis and purification technology of S-AMP using enzymatic reaction, and a small-scale synthesis system of S-AMP was established by using PhAdSS, which laid the foundation for the establishment of the industrial production process of S-AMP

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Embodiment Construction

[0015] A method for preparing adenyl succinic acid or salt, using the Escherichia coli expression system pET-28-a vector to express PhAdSS with the sequence of MGSSHHHHHHSSGLVPRGSH at the N-terminal, and using the recombinant protein as a catalyst to catalyze inosinic acid IMP , L-aspartic acid and guanosine triphosphate GTP to synthesize S-AMP and GDP. After the synthesis reaction, use silica gel thin-layer chromatography to analyze the components in the product, and perform silica gel column chromatography separation according to the results of silica gel thin-layer chromatography Purify S-AMP, and then use recrystallization method to further improve the purity of product S-AMP.

[0016] The specific preparation method is:

[0017] (1) Construction of expression plasmid: In the previous study, we used the pET-19-b prokaryotic expression vector to express and purify PhAdSS. According to the reported expression and purification conditions of PhAdSS, 5 mg of PhAdSS with a purit...

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Abstract

The invention belongs to the technical field of scale production of candidate medicines for treating glucose-lipid metabolism disorder diseases. In order to solve the problems that existing S-AMP is difficultly produced, the price of the S-AMP is expensive, and raw material sources of the druggability of the S-AMP is difficulty ensured, the invention provides a preparation method of adenylosuccinic acid or adenylosuccinate. PhAdSS of which the N terminal is provided with an His-tag sequence is expressed by utilizing a pET-28-a carrier; with the PhAdSS as a catalyst, inosinic acid IMP, L-aspartic acid and guanosine triphosphate (GTP) are catalyzed and subjected to reaction to synthesize S-AMP; after the reaction is carried out, components in a product are analyzed by adopting silica gel thin-layer chromatography, and separation and purification on the S-AMP are carried out by adopting silica gel coluumn chromatography. The content of the s-AMP in a synthetic product is 61 percent, and arecycling ratio is about 20 percent. A preparation process is simple, the cost is low, and a basis is laid for building an industrialized S-AMP synthetic process; monopoly sale on S-AMP by foreign sellers is expected to be broken through, and the S-AMP druggability research cost can be greatly reduced.

Description

technical field [0001] The invention belongs to the technical field of large-scale production of candidate drugs for the treatment of disorders of glucose and lipid metabolism, and in particular relates to a preparation method of adenylosuccinic acid or a salt thereof and an application thereof. Using enzymes from deep-sea archaea to catalyze the synthesis of a natural adenosine compound: adenylyl succinic acid (salt), a small-scale laboratory preparation method was established. This compound has the function of promoting insulin secretion, and can be used as a candidate drug for improving glucose and lipid metabolism disorders to carry out druggability research. Background technique [0002] Adenylosuccinic acid (Adenylosuccinate), referred to as S-AMP] is a natural adenosine compound that promotes insulin secretion and restores the normal function of islet cells in patients with type II diabetes [1]. In previous studies, we found that S-AMP is an agonist of AMPK, and its ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/40C12N9/00C12N15/70C07H19/20C07H1/06C12R1/19
CPCC07H1/06C07H19/20C12N9/93C12N15/70C12P19/40C12Y603/04004
Inventor 谢勇王楠赵京凤成钟赵晓宏杨润梅吴崇明郭鹏蔡大勇薛健
Owner INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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