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Vespa magnifica peptide WVD-I, and preparation method and applications thereof

A technology of WVD-I and vespa, which is applied in the field of vespa peptide WVD-I and its preparation, and can solve problems such as toxic side effects and the like

Inactive Publication Date: 2020-05-08
DALI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the drugs currently used to treat stroke are chemical agents, which have problems of toxicity and side effects

Method used

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  • Vespa magnifica peptide WVD-I, and preparation method and applications thereof
  • Vespa magnifica peptide WVD-I, and preparation method and applications thereof
  • Vespa magnifica peptide WVD-I, and preparation method and applications thereof

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preparation example Construction

[0024] The present invention also provides a preparation method of Vespa peptide WVD-I described in the above scheme, comprising the following steps:

[0025] 1) mixing and dissolving the freeze-dried powder of Vespa venom gland secretion with water, centrifuging, and taking the supernatant to obtain a toxin solution;

[0026] 2) Put the toxin solution described in step 1) on a cation exchange chromatographic column for the first purification, collect the eluent corresponding to the eluent corresponding to the eluent peak that appears in 0 to 5 min, and freeze-dry the eluent to obtain the first Freeze-dried powder, mixing and dissolving the first freeze-dried powder and double distilled water to obtain the first purified product;

[0027] 3) Put the first purified product described in step 2) on an anion-exchange chromatographic column for second purification, collect the eluent corresponding to the eluent corresponding to the eluent peak that appears in 32 to 35 minutes, and ...

Embodiment 1

[0037] Example 1 Separation, purification and mass spectrometry analysis of WVD-I

[0038] 1. Experimental materials and methods

[0039] 1.1 Separation, purification and mass spectrometry analysis of WVD-Ⅰ

[0040] (1) Freeze-dried powder of Vespa venom secretion (self-made: the Vespa venom secretion, that is, the Vespa venom containing impurities such as dust, was purchased from the Vespa farmers; The product that is obtained, "freeze-dried powder of Vespa venom secretion" contains 60% polypeptide and protein) is dissolved in double-distilled water and made into a 5mg / mL toxin solution, centrifuged at 10000rpm for 5min, and the supernatant uses a disposable filter (0.22 μm) and stored at 4°C.

[0041] (2) The first step of separation was carried out by cation-exchange HPLC (Agilent Bio WCX) for loading the crude virus. W2998 detector detection, the detection wavelength is 280nm, and the column temperature is 25°C. The mobile phase was a four-phase elution system with a f...

Embodiment 2

[0054] Embodiment 2 makes coating agent to the WVD-I that embodiment 1 obtains

[0055] 1. Instruments and Reagents

[0056] Instruments: constant temperature water bath, beaker (25mL, 50mL, 100mL), measuring cylinder, stopwatch, measuring cup (5mL, 10mL), glass plate, glass rod.

[0057] Reagent: polyvinyl alcohol PVA-124, manufacturer: Guangzhou Qisheng Chemical Co., Ltd.; glycerin; 95% ethanol.

[0058] Raw drug: WVD-I prepared in Example 1.

[0059] 2. Preparation method

[0060] 2.1 Take PVA-124, add 12mL of water to 1g, add 24mL of water to 2g, add 36mL of water to 3g, soak for 24h, then heat in a water bath at 85°C for 10-20min until the PVA-124 is completely dissolved, cool to obtain liquid A, and set aside.

[0061] 2.2 Take another 50mL beaker, add 2.4g of glycerin, measure 9.6mL of 95% ethanol (25% alcohol content) with a measuring cylinder, add it, and stir thoroughly to obtain liquid B.

[0062] 2.3 Before use, add liquid A of the water phase into liquid B of ...

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Abstract

The invention provides vespa magnifica peptide WVD-I, and a preparation method and applications thereof, and belongs to the technical field of biochemical drug preparation. The lyophilized powder of poison gland secretions of vespa magnifica is taken as a raw material; the WVD-I can be obtained through three-step separation of cation exchange high performance liquid chromatography, anion exchangehigh performance liquid chromatography and reversed phase high performance liquid chromatography; and purity can reach more than 98% through mass spectrum identification, and molecular weight is about1,563 Da.

Description

technical field [0001] The invention relates to the technical field of preparation of biochemical drugs, in particular to a vespa peptide WVD-I and its preparation method and application. Background technique [0002] Biotoxins are chemical substances with the highest biological activity and toxicity, and are of great value in exploring the process of life movement and developing new drugs. [0003] Stroke is an acute cerebrovascular disease, which is a group of diseases that cause brain tissue damage due to sudden rupture of blood vessels in the brain or blockage of blood vessels that prevents blood from flowing into the brain, including ischemic and hemorrhagic strokes. According to the survey, stroke is the leading cause of disability among adults in China. Stroke has the characteristics of high morbidity, high mortality and high disability rate. Most of the drugs currently used to treat stroke are chemical agents, which have problems of toxicity and side effects. Cont...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K1/18C07K1/20A61K38/10A61P9/10A61P25/00
CPCA61K38/00A61P9/10A61P25/00C07K7/08
Inventor 李辉魏永凯王朋川张汉超杨志斌赵昱
Owner DALI UNIV
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