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Small molecule peptides and their uses

A technology for small molecule peptides and uses, applied in the direction of peptides, medical preparations containing active ingredients, peptide/protein components, etc., to achieve the effects of easy large-scale production, good α-glucosidase inhibitory activity, and obvious application potential

Inactive Publication Date: 2020-06-30
田丽
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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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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] A small molecular polypeptide, the amino acid sequence of the small molecular polypeptide is shown in SEQ ID No.1;

[0016] SEQ ID No. 1: DDCCKVKQKSSSKKLDL.

[0017] The isoelectric point of the small molecule polypeptide is 9.0.

[0018] The molecular weight of the small molecule polypeptide is 1925.26 g / mol.

[0019] The preparation method of the small molecule polypeptide:

[0020] Choose amino acid-King resin as the carrier (resin), fully swell the resin with dichloromethane, add the first Fmoc-protected amino acid, wash it several times with dimethylformamide, and use an appropriate concentration of DBLK to remove the Fmoc-protecting group Then, wash it several times with dimethylformamide, wash away DBLK, weigh the appropriate condensing agent benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate and activate Reagent methylmorpholine and the second Fmoc-protected amino acid (Fmoc-Asp-OH) at the C-terminal (Fmoc-Asp-OH) were coupled, detected by ninhydr...

Embodiment 2

[0022] Determination of α-glucosidase activity of small molecule polypeptides in vitro.

[0023] 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.

[0024] (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).

[0025] (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.

[0026] (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.

[0027] (4) Preparation of stop solution: Sodium carbonate (Na2CO3) was purchased from Shanghai...

Embodiment 3

[0036] Prepare 2,2-azino-bis(3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt (ABTS) into a 2mM solution with distilled water, mix 50mL of the above solution with 200mL K2S2O8 aqueous solution (70mM) , placed in the dark for 12-16h, to get 2,2-azino-bis(3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt free radical ion (ABTS· + ) solution. ABTS· with phosphate buffered saline (PBS) + The solution was diluted to an absorbance of 0.70±0.02 (Abs=734nm). Take 0.6mL peptide in DMSO solution (0.2,0.4,0.6,0.8μM), add 4mL ABTS· + solution, and measure its absorbance at 5 min. ABTS · + The calculation formula of free radical scavenging rate (S%) is as follows: S%=(A 0 -A) / A 0 , where A 0 for ABTS· + The absorbance of the solution, A is the absorbance after adding the drug solution.

[0037] Table 1. Peptides on ABTS· + The scavenging rate of free radical ions

[0038] Concentration (μM) clearance rate(%) 0.2 26.8 0.4 51.2 0.6 70.3 0....

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Abstract

The invention discloses a micro-molecule polypeptide, and the amino acid sequence of the micro-molecule polypeptide is as shown in the SEQ ID NO.1. The invention further provides application of the micro-molecule polypeptide. The polypeptide is excellent in alpha-glucosidase inhibitory activity and antioxidant activity and strong in stability, and has a remarkable application potential, and the structure of the polypeptide is easy to regulate and control.

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 Patents(China)
IPC IPC(8): C07K7/08A61K38/10A61P3/10A61P39/06
CPCA61K38/00C07K7/08
Inventor 李培源苏炜霍丽妮陈睿
Owner 田丽
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