Preparation and application of ricin b

A protein, type 2 diabetes technology, applied in the field of anti-diabetic protein drugs, can solve problems such as many side effects and unsatisfactory effects, and achieve the effects of lowering blood sugar and blood lipids, and improving body weight and water intake

Active Publication Date: 2020-10-09
海木集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the problem of unsatisfactory effects and many side effects of current drugs for treating type 2 diabetes, the present invention provides a recombinant ricin B chain protein mutant and its expression method, and verifies its hypoglycemic effect on type 2 diabetes rats

Method used

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  • Preparation and application of ricin b
  • Preparation and application of ricin b
  • Preparation and application of ricin b

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Preparation of recombinant ricin B chain protein mutant

[0024] 1 Obtaining of E. coli expression engineering bacteria BL21(DE3) / PET28a-tRTB

[0025] Under the condition of unchanged amino acid sequence, codon optimization is performed according to the existing ricin B chain (rRTB) mutants, and the full sequence of ricin B chain codon optimized according to the above-mentioned ricin B chain is used as a template, using tRTB sense primer PCR amplification is performed with tRTB anti-sense primer, and the obtained PCR product is ligated to the pMD19-T simple cloning vector (Bao Bioengineering (Dalian) Co., Ltd.) to obtain pMD19T-tRTB. Link with the PET-28a expression vector that has been cut with the same NdeI and HindIII restriction enzymes. The resulting link product is transferred to E. coli competent cells BL21 (DE3). The positive bacteria are screened and the plasmids are extracted and sequenced. The result is The plasmid is a plasmid obtained by inserting seq...

example 2

[0029] Example 2 Experiment of the effect of recombinant ricin B-chain protein mutant on type 2 diabetic rats

[0030] 1 Experimental animals and groups

[0031] 1.1 Model making

[0032] Ten male wistar rats used in the experiment were randomly selected as normal control group. After other rats were fed with high-fat diet for 4W, the remaining single small dose (30mg / kg) intraperitoneal injection of STZ induced type 2 diabetes model (STZ dissolved in In citric acid-sodium citrate buffer, the injection is completed within half an hour), after 1W, fasting but water for 12 hours, blood is collected from the tail vein, and fasting blood glucose (FBG) is measured with a blood glucose meter, where FBG≥7.8mmol / 1 is a successful type 2 diabetes model. The test animals were fed in a standardized feeding room with alternating light and dark for 12 hours in the animal center. The room temperature was 20-25℃, the relative humidity in the room was 45%-60%, and the feeding room was well ventil...

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Abstract

The present invention relates to the technical field of antidiabetic drugs, and specifically to a recombinant protein drug for treating type 2 diabetes. The drug can reduce blood glucose and serum lipid levels in type 2 diabetic rats.

Description

Technical field [0001] The present invention relates to the technical field of anti-diabetic protein drugs, in particular to a recombinant ricin B chain protein mutant for type 2 diabetes. Background technique [0002] Diabetes is an immune system disease that is caused by insufficient insulin secretion and insulin resistance. It involves various systems throughout the body and can even induce many fatal complications. It seriously harms human health. Diabetes has become a current The third major disease endangering human health. More than 90% of diabetic patients have type 2 diabetes (type 2diabetes mellitus, T2DM). Currently, about 246 million people worldwide suffer from T2DM, and it is expected that the number of T2DM patients will increase to 380 million within 20 years. China is the country with the largest number of diabetes patients in the world, but the blood glucose compliance rate of T2DM patients in China is seriously insufficient. At this stage, the drugs used to ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/16A61P3/10
CPCA61K38/168A61P3/10A61K38/16C07K14/415
Inventor 刘文森许娜侯峰曹利利宫鹏涛李思明陈星远丁鹤王典
Owner 海木集团有限公司
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