Chiral medicine resolution medium based on polypeptide combined chiral recognition base and preparation method thereof

A chiral drug and chiral recognition technology, applied in chemical instruments and methods, and other chemical processes, can solve the problems of surface bonding, small load of stable samples, and unclear mechanism of action, etc. The effect of split range, high bonding density and uniform surface distribution

Inactive Publication Date: 2012-09-26
WUHAN HUISHITONG BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently commercial protein chiral chromatographic stationary phases are mainly albumin (such as HSA) and α-acid glycoprotein (α-AGP), which can resolve many kinds of chiral compounds, but their mechanism of action is still unclear; In addition, the amount of surface bonding, stability and sample loading of this type of chiral stationary phase are small

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Example 1: Take 2~10g OV (ovalbumin) and dissolve it in a buffer solution with pH 8.1, add 5% trypsin solution (pH 3.0) until the total solution contains trypsin equivalent to 0.5%, and dissolve it at 25~ Carry out enzymatic hydrolysis at 36°C for 5-30 minutes, and remove the enzyme protein from the polypeptide combination library obtained after enzymatic hydrolysis by adjusting the temperature. Then take 10g of amino-modified porous silica gel with a pore size of 8nm and a particle size of 5mm, first react with carbonyl imidazole (CDI) in acetonitrile, filter and wash after the reaction, and then mix with the above solution for removing enzyme protein (adjust pH 7.0) React at 60-90°C for 2-10 hours, filter the reaction product, wash with ethanol several times, and dry to obtain the desired resolution medium. The resolution medium has a carbon content of 8.0%.

Embodiment 2

[0012] Example 2: Take 2~10g OV (ovalbumin) and dissolve it with a pH8.5 buffer solution, add 1% elastase solution (pH8.5) to the total solution containing elastase equivalent to 0.1%, and dissolve it at 28~ Carry out enzymatic hydrolysis at 37°C for 5-30 minutes, and then remove the enzyme protein from the polypeptide combination library obtained after enzymatic hydrolysis by adjusting the temperature. Then take 10g of amino-modified porous silica gel with a pore size of 10nm and a particle size of 10mm, and treat it with a similar treatment method as in Example 1, and then react with the above-mentioned enzyme-removing solution (adjust the pH to 7.0) at 60-90°C for 2 After ~10 hours, the reaction product was filtered and washed several times with ethanol, and then dried to obtain the desired resolution medium. The resolution medium has a carbon content of 8.4%.

Embodiment 3

[0013] Embodiment 3: Get 2 ~ 10gBSA (bovine serum albumin) and dissolve it with the buffer solution of pH8.1, add 5% trypsin solution (pH3.0) to contain the trypsin equivalent to 0.5% in the total solution, at 25 Carry out enzymatic hydrolysis at ~36°C for 5-30 minutes, denature heavy metals to remove enzymes, filter, add elastase to the filtrate, which is equivalent to about 0.1% of the total solution, enzymolyze at 28-37°C for 5-15 minutes, and then use enzyme The peptide combinatorial library obtained after digestion was denatured and filtered to remove enzyme proteins. Then take 10g of amino-modified porous silica gel with a pore size of 8nm and a particle size of 30mm, and treat it similarly to Example 1, then react with the above-mentioned enzyme-removing protein solution (adjust the pH to 7.0) at 60-90°C for 2-10 hours , the reaction product was filtered and washed several times with ethanol, and dried to obtain the desired resolution medium. The resolution medium has ...

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Abstract

A chiral medicine resolution medium based on a polypeptide combined recognition base is obtained by bonding the polypeptide combined base with different chiral recognition capabilities onto the surface of porous silica gel with its pore size being 8-12nm and particle size being 5-30mm through chemical modification. According to a preparation method of the above chiral medicine resolution medium, one or some medicine-related proteins undergo enzymatic hydrolysis at 25-37 DEG C by the use of trypsin or papain or elastase or one or some enzymes thereof; zymoprotein is removed from the polypeptide combined base obtained after enzymatic hydrolysis by denaturation and adsorption or dialysis, and then the polypeptide combined base is bonded onto the porous silica gel surface by chemical modification so as to obtain the required chiral medicine resolution medium. The prepared chiral medicine resolution medium has wide chiral resolution range and uniform surface distribution, and its bonding density is higher than the bonding density of a protein stationary phase. The chiral medicine resolution medium has the combined chiral recognition base which can be used for resolution and analysis of a plurality of chiral medicines and provides a foundation for developing chiral resolution materials with a wider resolution range and chiral medicines with better specificity.

Description

technical field [0001] The invention relates to a chiral drug resolution medium based on a polypeptide combination recognition library and a preparation method thereof. Background technique [0002] The high-performance liquid chromatography based on chiral stationary phase (Chiral stationary phase, CSP) can be widely applied to various compounds, and is suitable for the analysis and determination of routine and biological samples. The preparation and separation are fast and convenient, and the reliability of quantitative analysis is high. , the number of compounds studied by this method has reached as many as thousands, and it has become one of the most important ways at present. There are many types of chiral stationary phases used for chiral separation in liquid chromatography, such as Proteins, Polysaccharides, Chiralpolymers, Crownethers, and cyclodextrins. ), ligand exchange type (containing amino acid residues or other groups that can complex with ions like metal cop...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/29B01J20/30
Inventor 宋文彩刘武邱华
Owner WUHAN HUISHITONG BIO ENG
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