Separation method of palmitoyl pentapeptide-4

A technology of palmitoyl pentapeptide and glucan gel, which is applied to the preparation methods of peptides, chemical instruments and methods, peptides, etc., can solve the problems of large consumption of organic solvents, cumbersome steps, low efficiency, etc. The effect of high adsorption capacity and improved efficiency

Active Publication Date: 2022-04-15
浙江湃肽生物股份有限公司深圳分公司
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  • Description
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  • Application Information

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Problems solved by technology

At present, the separation and purification methods of polypeptide mainly include: salting out method, gel filtration method, ion exchange chromatography method, adsorption chromatography method, isoelectric point precipitation method, enzymatic hydrolysis method, etc., but the steps of traditional methods are cumbersome and inefficient, requiring Consumption of a large amount of organic solvents, resulting in great waste
In order to make up for the shortcomings of the traditional method, Liu Shaojun and others proposed a purification method of palmitoyl pentapeptide-4, which removes the impurities visible in the chromatography through reverse-phase polymer chromatography, and then converts it into acetate by ion exchange chromatography, reducing the organic The use of solvents, but the purity of the purified palmitoyl pentapeptide-4 is only 97.51%, the purity is low

Method used

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Examples

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Embodiment 1

[0072] This embodiment provides a method for preparing modified dextran gel microspheres, the method comprising the following steps:

[0073] 1) Dissolve dextran with a molecular weight of 40,000 g / mol in deionized water and stir evenly according to the ratio of material to liquid 1g:8mL, add sodium periodate with 0.45 times the mass of dextran in an ice-water bath, and react in the dark at room temperature for 3.5 h, dialyzed, and freeze-dried to obtain aldylated dextran;

[0074] 2) Mix the formylated dextran obtained in step 1), sodium chloride, and deionized water according to the mass ratio of 1:0.26:3.8, stir evenly at room temperature, and add o-sulfonic acid 0.32 times the mass of the formylated dextran p-Phenylenediamine, stirred for 8 minutes, stirred at 65°C for 25 minutes to obtain dextran gel;

[0075] 3) Mix Sephadex and 20wt% isopropanol aqueous solution at a mass ratio of 1:3, stir and swell for 40 minutes, add 2-(3-methoxyphenyl)isonicotinic acid 3.2 times th...

Embodiment 2

[0078] This example provides another preparation method of modified dextran gel microspheres, the steps are basically the same as in Example 1, the difference is that in step 3) 2-(3-methoxyphenyl) The amount of isonicotinic acid added is 1.5 times the mass of Sephadex.

Embodiment 3

[0080] This example provides another preparation method of modified dextran gel microspheres, the steps are basically the same as in Example 1, the difference is that in step 3) 2-(3-methoxyphenyl) The amount of isonicotinic acid added is twice the mass of Sephadex.

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Abstract

The invention provides a method for separating palmitoyl pentapeptide-4, which belongs to the technical field of polypeptide separation, and specifically includes: 1) placing the crude palmitoyl pentapeptide-4 in a dichloromethane / methanol mixed aqueous solution for microwave pretreatment, filtering 2) separation and purification by gel permeation chromatography; 3) separation and purification by reversed-phase high performance liquid chromatography twice; 4) freeze-drying to obtain high-purity palmitoyl pentapeptide-4. The palmitoyl pentapeptide-4 crude product obtained by solid-phase synthesis is separated and purified by the above method, the separation speed is fast, the sensitivity is high, and the resolution is strong, and most of the side reaction products and organic solvents in the crude peptide can be removed to obtain High-purity palmitoyl pentapeptide-4, with high separation and purification efficiency.

Description

technical field [0001] The invention belongs to the technical field of polypeptide separation, and in particular relates to a separation method of palmitoyl pentapeptide-4. Background technique [0002] Palmitoyl Pentapeptide-4 is one of the earliest and most widely used multi-peptides in the peptide series. The mechanism of action of palmitoyl pentapeptide-4 is: by acting on dermal fibroblasts, it promotes the synthesis of macromolecular substances such as collagen types I, III, and IV, fibronectin and glycosaminoglycans, and repairs and rebuilds the extracellular matrix And the epidermis-dermis junction to achieve the effect of improving wrinkles. At the same time, palmitoyl pentapeptide-4 can also promote the expression of wound repair genes, which is beneficial to the recovery of skin tissue. The anti-wrinkle activity of palmitoyl pentapeptide-4 is higher than that of vitamin A and vitamin C, and it has no side effects. Therefore, it is sought after by the cosmetics ind...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/16C08J3/12C08J3/24C08L5/02C08B37/02C07K7/06C07K1/20
CPCC08J3/12C08J3/24C08B37/0006C07K7/06C07K1/20C07K1/16C08J2305/02
Inventor 陈晓航陈超虞慧飞周永兵
Owner 浙江湃肽生物股份有限公司深圳分公司
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