Drug having weight-loss function

A drug and application technology, applied in the field of GLP-1 mutants, can solve problems such as irritation and allergic reactions, physical and psychological pain of patients, and inconvenient use and carrying, so as to reduce degradation, continuously control body weight, and avoid painful effect

Active Publication Date: 2019-09-20
福州市台江区希吉亚健康科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The way of injection is inconvenient to use and carry, the drug compliance is poor, and there may be risk

Method used

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  • Drug having weight-loss function
  • Drug having weight-loss function
  • Drug having weight-loss function

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 The transformation of the original sequence of GLP-1 in the present invention

[0032] Starting from the original sequence of GLP-1, the mutant polypeptide sequence is modified and designed, and a polypeptide product with a purity of up to 85% is obtained through chemical synthesis (Zhongpeptide Biochemical). The peptide sequence is as follows:

[0033] GLP-1 Fragment (7-37aa): HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG

[0034] GM (SEQ ID NO: 1): HHEGTFTSDVSSYLEGQAAKKFIAWLVRGGKKKKKYGRKK RRQRRREF

[0035] GM is based on the original sequence of GLP-1, mutating the 8th alanine of GLP-1 to histidine, the 27th glutamic acid to lysine, and the 34th lysine to arginine acid, arginine at position 36 is mutated to glycine, glycine at position 37 is mutated to lysine, and a sequence is added at the end.

[0036] The differences between the original sequence of GLP-1 and the mutant GM are shown in Table 1.

[0037] Table 1 Sequence comparison of GLP-1 and mutant GM

[0038...

Embodiment 2

[0041] Embodiment 2 Mutant GM transforms Escherichia coli Nissle1917

[0042] 1. Add a signal peptide sequence before the sequence of GLP-1 mutant GM, add HIV membrane-penetrating peptide sequence and 6 His-tag sequences at the 3'-end, and construct GLP that can be secreted and expressed and has the ability to pass through the cell membrane -1 mutant, its nucleotide sequence is shown in SEQ ID NO:2.

[0043] SEQ ID NO: 2:

[0044] atgaaaaagaacatcgcattcctcctggcatctatgtttgttttctctatcgctaccaacgcttacgctggatcccaccacgagggcaccttca cctccgacgtgtcctcctacctggagggccaggccgccaagaagttcatcgcctggctggtgcgcggcggcaagaagaagaagaagt acggccgcaagaagcgccgccagcgccgccgcctcgaggacgacgacgacaagcaccatcaccatcaccattaa

[0045] 2. Cloning the nucleotide sequence shown in SEQ ID NO: 3 into the BamHI and SalI sites of the Escherichia coli Nissle1917 expression vector pGEX to obtain the pGEX-GLP-1GM vector, and transform it into E. coli E.coli BL21(DE3 ), the target fragment size is about 12KD. After purificatio...

Embodiment 3

[0047] The preparation of embodiment 3 mutant GM transformation Escherichia coli Nissle1917 preparation

[0048] 1. Preparation of mutant GM transformed Escherichia coli Nissle1917 oral solution

[0049] After adding erythromycin-resistant LB liquid medium with a final concentration of 200 μg / ml for steam sterilization, Escherichia coli Nissle1917 (prepared in Example 2), which was stably transformed with a gene identified by PCR verification and induction expression experiment, was inoculated at 37° C. Cultivate on a shaker at 250r / min until the OD600 value reaches 0.8-1.0, then centrifuge for 5 minutes to harvest the bacteria, wash them twice with normal saline, collect the bacterial precipitate by centrifugation, and add L-arabinose solution with a final concentration of 2% (w / v) Or 20% fructooligosaccharide solution to resuspend the bacteria, prepared to be not less than 10 per milliliter 8 The fresh bacterial solution of viable bacteria was stored in a refrigerator at 4°...

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Abstract

The invention discloses a use of a GLP-1 mutant in preparation of weight-loss drugs. The GLP-1 mutant is obtained by mutation of amino acid sites on the basis of wild-type GLP-1, and has an amino acid sequence shown in SEQ ID NO:1. The GLP-1 mutant can effectively reduce body weight, fat-body ratio and other indicators, can be used for reducing body weight of obesity people, and has broad application prospects.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and specifically relates to the use of GLP-1 mutants. Background technique [0002] With the changes in people's dietary structure and lifestyle, obesity has become a worldwide epidemic. The prevalence of overweight and obesity worldwide is increasing year by year. Especially in my country, the incidence of overweight and obesity has approached or even reached the initial level of developed countries in recent years. For those with a high BMI index, obesity will lead to an increase in the incidence of other diseases, the most notable of which are diabetes, cardiovascular disease and cancer. Overweight / obesity seriously endangers human health. [0003] At the same time, overweight / obesity is an independent risk factor for the occurrence and development of Type 2 Diabetes Mellitus (T2DM), and the two are disease states that accompany each other. Studies have shown that every 1kg / m ...

Claims

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Application Information

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IPC IPC(8): A61K38/26A61K35/74A61K35/744A61P3/04A23L33/00
CPCA61K38/26A61K35/74A61K35/744A61P3/04A23L33/00A23V2002/00A23V2200/332Y02A50/30
Inventor 顾建文马婕马永平
Owner 福州市台江区希吉亚健康科技有限公司
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