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Buthotoxin polypeptide and preparation method thereof

A technology for preparing scorpion venom and column, which is applied in the field of scorpion venom polypeptide and its preparation, can solve the problems of no specific qualitative and quantitative indicators, purity, unstable quality, and difficult determination of fraction composition, etc., and achieve fast purification speed, convenient transportation and Effect of storage and quality control

Active Publication Date: 2005-10-26
沈阳成泰实业有限公司
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AI Technical Summary

Problems solved by technology

In the patent application documents disclosed by Chinese patent application numbers 93109133, 97120965, and 98101335, scorpion venom injections and preparation methods thereof are disclosed. The disadvantage of the preparation of scorpion venom injection is that the raw materials of scorpion venom in the scorpion venom injection are all in the form of scorpion venom protein or polypeptide mixture, and its purity and quality are not stable enough, which directly affects the use effect; , quantitative indicators, it is difficult to meet the national requirements for the quality of pharmaceutical raw materials
The existing methods for the preparation of scorpion venom peptides mostly use gel chromatography column method, which has the disadvantages of slow separation speed and difficult determination of fraction composition

Method used

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  • Buthotoxin polypeptide and preparation method thereof
  • Buthotoxin polypeptide and preparation method thereof
  • Buthotoxin polypeptide and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The instrument is a high-performance liquid chromatography E-C2000 chromatographic data processing workstation produced by Dalian Elite Analytical Instrument Co., Ltd., equipped with two identical P230 high-pressure constant-flow pumps (A pump and B pump), provided by Dalian Elite Analytical Instrument Co., Ltd. Company production. The ultraviolet spectrophotometer used for detection is: UV-2800 ultraviolet visible spectrophotometer

[0045] The chromatographic column is a semi-preparative column, specification: 4.60mm (inner diameter) × 250mm (height);

[0046] The chromatographic column filler is the CM Sepharose series produced by Sweden Pharmcia Company, and the steps are:

[0047] (1) Start the high performance liquid chromatograph, open the EC2000 chromatographic data processing workstation, and set the chromatographic conditions, wherein the chromatographic conditions are set to:

[0048] Flow rate: 5~50ml / min; Pressure: 3Mpa; Detection wavelength: uv~210nm;

...

Embodiment 2

[0065] Instrument used, chromatographic condition, gradient program are set with embodiment 1,

[0066] Weigh 100mg of scorpion venom powder, dissolve it in 1ml of mobile phase B solution, inject the sample after dissolving, run pump B to punch the column for about 20 minutes, after the instrument is stable, add the prepared scorpion venom powder solution, press the start button to start the gradient Elution, collection of fractions: after sample injection, run the two pumps A and B according to the set gradient program, the flow rate is 5ml / min, and start collecting when the peak rises for about 5 minutes through the chromatographic workstation, and stop collecting when the peak falls, see the attached picture 1 Liquid chromatogram of scorpion venom polypeptide, the obtained liquid is colorless or light yellow clear liquid, the fraction collected is about 40-50ml, and detected by ultraviolet spectrophotometer 280 = about 0.600~0.700; the collected fractions are confirmed by e...

Embodiment 3

[0069] Instrument used, chromatographic condition, gradient program are set with embodiment 1,

[0070] Weigh 600mg of scorpion venom powder, dissolve it in 6ml of mobile phase B solution, inject the sample after dissolving, run the B pump to flush the column for about 20 minutes, after the instrument is stable, add the prepared scorpion venom powder solution, press the start button to start the gradient Elution, collection of fractions: after sample injection, run the two pumps A and B according to the set gradient program, the flow rate is 50ml / min, and observe the chromatographic workstation for about 1 minute when the peak rises and collects until the peak falls. See Figure 1 Scorpion venom polypeptide liquid chromatogram, the obtained liquid is a colorless or light yellow clear liquid, the collected fraction is about 160-250ml, and the A 280 = about 1.050~1.550; the collected fractions are confirmed by electrophoresis that its molecular weight is between 5000~10000 Dalton...

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Abstract

The present invention relates to a scorpion venom polypeptide and its perparation method, belonging to the field of biological pharmaceutical technology. The scorpion venom polypeptide extracted by using HPLC has the characteristics of strong activity, high purity, stable quality and controllable components, and has the good effect of stopping pain, resisting platelet aggregation and stopping drugs.

Description

technical field [0001] The invention relates to the technical field of biopharmaceuticals, in particular to a scorpion venom polypeptide and a preparation method thereof. Background technique [0002] In traditional Chinese medicine, scorpion has a long history of medicinal use. Scorpion has a mild taste and taste, and belongs to the Yin meridian of the foot. In recent years, more and more people pay attention to the research and application of scorpion venom. According to research, scorpion toxin is the main component of scorpion pharmacological effects, which has antiepileptic, antithrombotic, anti-inflammatory, anti-tumor, and strong central analgesic effects. Some have been used for disease treatment at present. In the patent application documents disclosed by Chinese patent application numbers 93109133, 97120965, and 98101335, scorpion venom injections and preparation methods thereof are disclosed. The disadvantage of the preparation of scorpion venom injection is th...

Claims

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Application Information

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IPC IPC(8): C07K14/435
Inventor 孟祥军赵恩成顾孝先赵婧婷
Owner 沈阳成泰实业有限公司
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