Polypeptide for preventing and treating diabetes, preparation method and application thereof

A technology of diabetes and site, applied in the field of biomedicine, can solve gastrointestinal discomfort and other problems, achieve the effect of improving sensitivity, simple extraction process, and restoring the function of islets

Inactive Publication Date: 2019-02-26
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The GLP-1 peptide drugs and their protectors and agonists that have been released in recent years are also injection dru

Method used

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  • Polypeptide for preventing and treating diabetes, preparation method and application thereof
  • Polypeptide for preventing and treating diabetes, preparation method and application thereof
  • Polypeptide for preventing and treating diabetes, preparation method and application thereof

Examples

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preparation example Construction

[0043] The preparation method for extracting and isolating the above four polypeptides from chickpeas comprises the following steps:

[0044] (1) Soak the chickpeas for germination, mash them, extract them with 0.1-0.2M acetic acid at 10-20°C for 10-12 hours, separate the slurry, and take the separation liquid;

[0045] (2) The separation liquid obtained in step (1) is filtered with a ceramic membrane, and the filtrate is filtered with an organic membrane, and after retaining the integral with a molecular weight of less than 5000 Daltons, freeze-drying;

[0046] (3) Using reversed-phase liquid chromatography (HPLC) to separate the above-mentioned polypeptide or a mixture thereof from the freeze-dried product in step (2).

[0047] The preparation method of the chickpea four kinds of amino acid polypeptides provided by the invention is suitable for industrial production, the operating conditions are easy to control, and the product quantity and quality are stable.

[0048] Sequ...

Embodiment 1

[0056] Extract 4 kinds of 37 amino acid polypeptides of chickpea of ​​the present invention according to the following method:

[0057] Take 1 kg of chickpea seeds, soak them for 12 hours, germinate them, mash them, stir them with 2L of 0.1M acetic acid solution, extract them at 10-20°C for 10 hours, separate the slurry and get the clear liquid. The supernatant was filtered with a ceramic membrane, and the filtrate was filtered with an organic membrane, and after the integral with a molecular weight of 5000 Daltons (DA) and less than 5000 Daltons was retained, it was freeze-dried. Finally, about 10 grams of product were obtained by preparative reversed-phase liquid chromatography (HPLC).

[0058] Collect the separated liquid of each separation peak in HPLC (see figure 1 ) conducted pharmacodynamic tests respectively, and found that several peaks appearing in the time interval from 30.264 to 35.360 minutes had a hypoglycemic effect, and then carried out protein sequencing on t...

Embodiment 2

[0075] This example provides a composition for the treatment of diabetes prepared by using the chickpea 4 kinds of 37 amino acid polypeptides prepared in Example 1 and citric acid and lactic acid according to a certain ratio. It is a liquid dosage form, called a mixed potion. Percentage by weight, the 4 kinds of 37 amino acid polypeptides of chickpea accounted for 5%, citric acid accounted for 5%, and lactic acid accounted for 6% in the mixed medicine. There were 40 mice with type 1 diabetes and 40 mice with type 2 diabetes respectively. Saline and normal insulin were used as controls. The administration effect of the mixed medicine on type 1 diabetic mice and type 2 diabetic mice was tested, and the experimental results are shown in Table 4 and Table 5.

[0076] The numbers in the table represent the blood glucose concentration, and the administration methods are: insulin subcutaneous injection, mixed medicine and normal saline gavage. Normal saline was 100 μL / mouse, and insu...

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Abstract

The invention discloses chickpea four-type 37-amino acids polypeptides, a preparation method thereof and an application thereof in preparing a medicine for treating diabetes or a functional food. Eachpeptide is composed of 37 amino acids, the six cysteines in the molecule and the formed three pairs of disulfide bridges are highly conserved, and the three pairs of disulfide bonds form a cysteine knot domain. The chickpea four-type 37-amino acids polypeptides can be dissolved in polar solvents such as water, which are stable to heat and resistant to hydrolysis of protease in human digestive tract. The preparation cost is lower than that of insulin and other peptide drugs; the product is not only can be used for treating symptomatic doeases, but also treats the root, and can be used for recovering the quality and function of diseased pancreas at varying degrees; the product reduces or eliminates the body's insulin resistance, can be developed to oral dosage form, and is easier to use than insulin and other peptide drugs; while in use, no risk of hypoglycemia is generated; and the product is derived from edible plant chickpeas and has no toxic side effects.

Description

technical field [0001] The invention belongs to the field of biomedicine, and more specifically relates to a polypeptide compound for preventing and treating diabetes, its preparation method and application. Background technique [0002] Diabetes (diabetes) is a disorder of sugar, protein and fat metabolism, which has a high incidence worldwide and has become the three "health killers" alongside cardiovascular and tumors. The prevalence rate in my country has reached 10%, and the total number of patients has exceeded 100 million, ranking first in the world. Complications caused by diabetes such as cardiovascular and cerebrovascular diseases, renal failure, blindness in both eyes and diabetic foot seriously threaten people's life and quality of life. So far, there are no drugs that can really treat diabetes. [0003] In 1980, the World Health Organization (WHO) divided diabetes into: insulin-dependent diabetes melitus (IDDM, also known as type 1, T1D), non-insulin-dependent...

Claims

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Application Information

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IPC IPC(8): C07K14/415C07K1/14C07K1/34C07K1/20A61K38/16A61P3/10
CPCA23L33/18A23V2002/00A61K38/00C07K14/415A23V2200/328A23V2250/55
Inventor 陈正望陈威顿新鹏张欣童宇星
Owner HUAZHONG UNIV OF SCI & TECH
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