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Biological modifying and recombinant human growth hormone compound and preparing method thereof

A technology of human growth hormone and complex, applied in the field of biological modification of peptide or protein and its preparation, can solve the problems of poor biological stability and short half-life (only about 30 minutes), and achieve improved biological stability and high possibility , the effect of fewer types of by-products

Inactive Publication Date: 2006-02-22
EAST CHINA NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Recombinant human growth hormone is an active protein that is easily degraded by proteases in the body, resulting in poor biological stability. At the same time, recombinant human growth hormone has a relatively small molecular weight of 21,000 Daltons and is easily filtered by the glomerulus. The half-life is short (only about 30 minutes), and long-term repeated injections are required. Generally, continuous administration is required for 3 months to 2 years, which brings pain and inconvenience to patients.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 One of the preparation methods of biomodified recombinant human growth hormone complex. In this example, the amino donor substance used for biomodification is polyethylene glycol alkylamine.

[0019] In the first step, 1.0 mg of recombinant human growth hormone was weighed and dissolved in 1 g of 100 mM Tris-HCl pH6.0 solution containing 5 mM EDTA to dissolve it;

[0020] In the second step, add 0.8 mg of polyethylene glycol alkylamine to the solution of the first step, and mix evenly, so that the molar ratio of recombinant human growth hormone to be modified and polyethylene glycol alkylamine in the mixed solution is 1: 5;

[0021] In the third step, add transglutaminase mTG to the mixture obtained in the second step, and mix evenly so that the final concentration is 0.1-1.0u / ml;

[0022] In the fourth step, the mTG-containing mixture obtained in the third step is reacted at 4-55° C. for 0.5-24 hours;

[0023] The fifth step, after the reaction is over, ta...

Embodiment 2

[0027] Example 2 The second preparation method of the biomodified recombinant human growth hormone complex. In this example, the amino donor substance used for biomodification is polyethylene glycol alkylamine.

[0028] In the first step, 1.0 mg of recombinant human growth hormone was weighed and dissolved in 1 g of 100 mM Tris-HCl pH6.0 solution containing 5 mM EDTA to dissolve it;

[0029] In the second step, add 8 mg of polyethylene glycol alkylamine to the solution of the first step, and mix evenly, so that the molar ratio of recombinant human growth hormone to be modified and polyethylene glycol alkylamine in the mixed solution is 1:50 ;

[0030] The third step to the seventh step is carried out according to the corresponding operation steps in embodiment 1;

[0031] In the eighth step, the collected modified complex absorption peak solution is concentrated and freeze-dried to obtain 0.27-0.32 mg of pure biologically modified recombinant human growth hormone complex PEG...

Embodiment 3

[0032] Example 3 The third preparation method of the biomodified recombinant human growth hormone complex. In this example, the amino donor substance used for biomodification is polyethylene glycol alkylamine.

[0033] In the first step, 1.0 mg of recombinant human growth hormone was weighed and dissolved in 1 g of 100 mM Tris-HCl pH6.0 solution containing 5 mM EDTA to dissolve it;

[0034] In the second step, add 16 mg of polyethylene glycol alkylamine to the solution of the first step, and mix evenly, so that the molar ratio of recombinant human growth hormone to be modified and polyethylene glycol alkylamine in the mixed solution is 1:100 ;

[0035] The third step to the seventh step is carried out according to the corresponding operation steps in embodiment 1;

[0036] In the eighth step, the collected modified complex absorption peak solution is concentrated and freeze-dried to obtain 0.18-0.22 mg of pure biologically modified recombinant human growth hormone complex PE...

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Abstract

The invention is a bio-modified recombinant growth hormone (rhGH) complex and preparing method, making glutamine residue-containing rhGH react with amino donor polyethylene glycol alkyl amine PEG or monomethoxypolyethylene glycol (mPEG) under the catalysis of transglutaminase (mTG), to make acylamide in gamma-bit of the glutamine residue and amino in primary amine bit of the amino donor form an amido bond, thus obtaining it, namely PEG-rhGH or Mpeg-rhGH. It has high biostability and long in vivo half life, applied to prepare permanent injection drugs or develop oral drugs.

Description

technical field [0001] The invention relates to a biologically modified recombinant human growth hormone complex and a preparation method thereof, belonging to the technical field of peptide or protein biological modifiers and their preparation. Background technique [0002] Human growth hormone (hGH) is a polypeptide hormone composed of 191 amino acids secreted by the anterior lobe of the human pituitary gland. People have obtained recombinant human growth hormone (rhGH) through genetic engineering technology. In 1985, the U.S. FDA approved rhGH as a drug for marketing, and China also approved it as a drug for marketing in 1998 ("Genetic Engineering Drugs" edited by Li Yuan, Chemical Industry Press, December 2002, first edition P3-5), mainly used for It can be used to treat spontaneous pituitary dwarfism in children, and it can also be used clinically to treat burns, fractures, trauma, bleeding ulcers, Turner syndrome, tissue necrosis, muscular dystr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/61C07K17/02
Inventor 吴自荣黄静刘晨卞慧芳
Owner EAST CHINA NORMAL UNIV
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