Micro-molecule polypeptide and application thereof

A technology for small molecular polypeptides and uses, which is applied in the fields of peptides, medical preparations containing active ingredients, peptide/protein components, etc., to achieve the effects of obvious application potential, simple synthesis method, and easy large-scale production.

Active Publication Date: 2018-05-22
玛士撒拉(上海)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The α-glucosidase inhibitors clinically used in the treatment of diabetes mainly include acarbose, voglibose, miglitol, etc., but these compounds have certain side effects

Method used

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  • Micro-molecule polypeptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] A small molecular polypeptide, the amino acid sequence of the small molecular polypeptide is shown in SEQ ID No.1;

[0017] SEQ ID No. 1: ICKKMMKKSTLLQDDIL.

[0018] The isoelectric point of the small molecule polypeptide is 9.7.

[0019] The molecular weight of the small molecule polypeptide is 2008.54g / mol.

[0020] The preparation method of the small molecule polypeptide:

[0021] Select amino acid-king resin as the carrier (resin), fully swell the resin with dichloromethane, wash several times with dimethylformamide, use DBLK of appropriate concentration to remove the Fmoc-protecting group, and then use dimethylformamide to remove the Fmoc-protecting group. The amide was washed several times to remove DBLK, and the appropriate condensing agent benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate, the activator methylmorpholine and the C-terminal Two Fmoc-protected amino acids (Fmoc-Pro-OH) are coupled, detected by ninhydrin detection method to ensure th...

Embodiment 2

[0023] Determination of α-glucosidase activity of small molecule polypeptides in vitro.

[0024] All tests were carried out with a Microplate reader ELX808TM microplate reader (BioTek, USA) at 37°C. The data analysis software uses Origin software for data processing, and uses acarbose as a reference substance.

[0025] (1) Preparation of inhibitor stock solution: the tested inhibitors were formulated into 10 mM DMSO solution (the DMSO solution of the polypeptide of Example 1).

[0026] (2) Preparation of enzyme stock solution: α-glucosidase was purchased from Sigma Company of the United States; 1 mg / mL was prepared respectively with phosphate buffer solution of pH=6.8.

[0027] (3) Preparation of substrate stock solution: p-nitrophenyl glucoside (PNPG) was used as substrate, purchased from Sigma Company; 10 mg / mL was prepared respectively with phosphate buffer solution of pH=6.8.

[0028] (4) Preparation of stop solution: Sodium carbonate (Na2CO3) was purchased from Shanghai...

Embodiment 3

[0038] Take 0.1 ml of the DMSO solution (0.2, 0.4, 0.6, 0.8 μg / ml) of the polypeptide of Example 1 and add it to 8 ml of 0.004% 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) solution . Absorbance was measured at the maximum wavelength (517nm) until equilibrium. The formula for calculating the clearance rate is as follows:

[0039] S%=(1-A 样品 ) / A 空白 ×100%. Among them, A 空白 Absorbance of DPPH solution without adding peptide DMSO solution, A 样品 is the absorbance of the DPPH solution added to the DMSO solution of the compound. The result is as figure 1 shown. It can be seen that the polypeptide molecule of the present invention has excellent antioxidant performance, and the scavenging ability for DPPH free radicals can reach more than 99.5% at a relatively low concentration.

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Abstract

The invention discloses a micro-molecule polypeptide, which has an amino acid sequence as shown in SEQ ID No.1. The invention also provides application of the micro-molecule polypeptide. The polypeptide provided by the invention has favorable alpha-glucosaccharase inhibitory activity and antioxidant activity, high stability, an easily-regulated structure and a remarkable application potential.

Description

technical field [0001] The invention relates to a small molecule polypeptide. More specifically, the present invention relates to a small molecular polypeptide having α-glucosidase inhibitory activity and antioxidant activity. Background technique [0002] Diabetes is a group of metabolic diseases characterized by hyperglycemia. Hyperglycemia is caused by defective insulin secretion or impaired biological action, or both. The long-term high blood sugar in diabetes leads to chronic damage and dysfunction of various tissues, especially the eyes, kidneys, heart, blood vessels, and nerves. In the mechanism of action of current hypoglycemic drugs, inhibiting the activity of α-glucosidase, which can catalyze the last step of digestion of starch or sucrose to produce carbohydrates, is considered to be an ideal way to control type II diabetes. Clinically used α-glucosidase inhibitors for the treatment of diabetes mainly include acarbose, voglibose, miglitol, etc., but these compo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61K38/10A61P3/10A61P39/06
CPCA61K38/00C07K7/08
Inventor 李培源苏炜霍丽妮陈睿
Owner 玛士撒拉(上海)生物科技有限公司
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