Method for purifying cobra venom and its products

A cobra venom and purification method technology, which is applied to the preparation method of peptides, chemical instruments and methods, medical preparations containing active ingredients, etc., can solve the problems of low product purity, complicated cobra venom purification process, and high cost

Inactive Publication Date: 2019-06-25
ZHEJIANG JINGXIN PHARMA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The technical problem to be solved by the present invention is that the purification process of the existing cobra venom is complicate

Method used

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  • Method for purifying cobra venom and its products
  • Method for purifying cobra venom and its products
  • Method for purifying cobra venom and its products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0204] Example 1: SPFF+MMC+low pressure inversion process

[0205] Step 1: Preprocessing

[0206] Get 20 grams of crude poison freeze-dried powder (Chinese cobra, Naja naja atra, purchased from Yiwu Chunfeng Snake Industry Co., Ltd.), its purity is 11.4% (measure with method 2, the result is as shown in table 1-1), add pH 800ml of 20mM PB buffer of 7.0, fully dissolved, centrifuged at 10000rpm for 20min at 4°C, and poured out the supernatant for later use.

[0207] Table 1-1

[0208] Peak number

Impurity 2

cobra neurotoxin

Impurity 1

retention time / min

21.606

22.199

24.720

relative retention time

0.973

1.000

1.113

Peak width / min

0.2813

0.3053

0.2361

Peak area / mAU×s

131.02

1812.87

176.67

Peak height / mAU

7.27

92.63

10.84

Peak area percentage / %

0.83

11.43

1.11

[0209] Step 2: SPFF separation

[0210] SPFF column (self-packing column: Borgron empty column ...

Embodiment 2

[0224] Example 2: SPFF+MMC+Low Voltage Inversion Process 2

[0225] On the basis of Example 1, the reversed phase filler was changed to NMSIL 50-100C8 silica gel matrix, non-uniform particle size, average particle size 50μm, pore size The yield is 74.32% and the purity is 98.67% (measured by method 2, the results are shown in Table 2-1), and the identification data are basically the same as those in Example 1.

[0226] table 2-1

[0227] Peak number

Embodiment 3

[0228] Example 3: SPFF+MMC+low pressure reverse phase process 3

[0229] On the basis of Example 1, the reversed-phase filler was changed to UNIPS 40-300, phenyl matrix, uniform particle size, particle size 40 μm, pore size The yield is 95.71% and the purity is 98.22% (measured by method 2, the results are shown in Table 3-1), and the identification data are basically the same as those in Example 1.

[0230] Table 3-1

[0231] Peak number

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PUM

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Abstract

The invention discloses a method for purifying cobra venom and its products. The invention provides a method for purifying cobra venom, which comprises the following steps: (1) performing cation exchange chromatography on a phosphate buffer containing the cobra venom to obtain an eluent A containing an active ingredient; wherein the phosphate buffer containing the cobra venom includes the cobra venom and a phosphate buffer solution; (2) performing complex mode chromatography on the eluent A containing the active ingredient to obtain an eluent B containing the active ingredient; and (3) desalting the eluent B containing the active ingredient to obtain the active ingredient. The purifying method has the advantages of simple process, easy industrialization, and low cost, and obtained producthas the advantages of high purity and low content of related substances.

Description

technical field [0001] The invention relates to a purification method of cobra venom and a product thereof. Background technique [0002] Cobra venom is a natural protein secreted by the venom glands of snakes, containing a variety of proteins, polypeptides, enzymes and other small molecular substances. As one of the main components of cobra venom, neurotoxin (cobrotoxin, referred to as NT) has a central analgesic effect similar to morphine, and is used for the analgesic treatment of intractable pain, malignant tumor pain and joint pain, but the analgesic effect is good. For morphine, it has a long duration, no addiction and drug resistance, and it can also get rid of morphine addiction, so it has a unique therapeutic effect in cancer pain and drug addiction, but the two have completely different biological characteristics. [0003] In 1965, Yang Zhenzhong et al. first isolated crystalline toxin from Taiwan cobra (Taiwan Cobra) through cation-exchange cellulose CM-cellulose...

Claims

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Application Information

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IPC IPC(8): C07K14/46C07K1/18C07K1/20C07K1/16A61K38/17A61P25/04
Inventor 徐苗焕张保安
Owner ZHEJIANG JINGXIN PHARMA
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