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Drosophila melanogaster polypeptide for promoting tissue repair as well as preparation method and application thereof

A tissue repair, Drosophila technology, applied in the field of active polypeptides, active polypeptides that promote tissue repair, can solve problems such as unclear functional components, and achieve the effects of improving skin beauty, promoting proliferation and migration, and good stability.

Active Publication Date: 2020-06-16
NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The applicant's previous research found that the fruit fly extract has a good effect on promoting the proliferation and migration of human immortalized epidermal cells (HaCat), but its functional components are still unclear

Method used

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  • Drosophila melanogaster polypeptide for promoting tissue repair as well as preparation method and application thereof
  • Drosophila melanogaster polypeptide for promoting tissue repair as well as preparation method and application thereof
  • Drosophila melanogaster polypeptide for promoting tissue repair as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1 The preparation method of Drosophila repair peptide, which comprises the following steps:

[0031] (1) Preparation of fruit fly extract:

[0032] Take crushed or not crushed fruit flies as a whole, add 5 times the amount of 70% ethanol solution, reflux extraction twice, each time for 30 minutes, centrifuge, take the supernatant, concentrate and dry to obtain the fruit fly extract.

[0033] (2) Extraction:

[0034] Disperse the fruit fly extract concentrate with 70% methanol solution, add about 1 / 10 volume of chloroform, shake fully, after standing for 4 hours, discard the chloroform layer, concentrate part of the methanol solution, and dry to obtain the methanol extract part of Drosophila .

[0035] (3) Strong oxygen ion exchange resin (SCX) separation:

[0036] The Drosophila methanol extraction part that step (2) obtains is redissolved with 10mM ammonium formate solution (containing 25% acetonitrile, pH 2.7~3), and the sample SCX small column is loaded...

Embodiment 2

[0039] Example 2 The sequence analysis of the Drosophila repair peptide, which comprises the following steps:

[0040] Take the Drosophila polypeptide prepared in Example 1, and use nano-LC-ESI-MS / MS to analyze the amino acid sequence of the polypeptide. The mass spectrometry conditions are: ESI source, scanning mode: positive ion mode, and the first-level full scan range of the mass spectrometer is m / z 300~2000, the separation width is 3Da; the tandem mass spectrometry adopts the data-dependent secondary mass spectrometry scanning mode of the primary mass spectrometry, and sequentially selects the five ions with the highest ion intensity in the primary mass spectrometry for collision-induced dissociation (CID) secondary mass spectrometry mass spectrum. According to the obtained MS / MS spectrum, the amino acid sequence of the Drosophila repair peptide is: Ile-Glu-Lys-Arg-Ile-Pro-Phe-Ser-His-Asp-Asp-Arg-Leu-Gly, and the mass spectrum is as follows figure 1 shown.

Embodiment 3

[0041] Example 3 Synthesis of Drosophila Repair Peptide

[0042]According to the amino acid sequence obtained in Example 2, the dipeptidyl peptidase-4 inhibitory peptide Ile-Glu-Lys-Arg-Ile-Pro-Phe-Ser-His-Asp-Asp-Arg- Leu-Gly, the purity of the synthesized polypeptide is 98% by HPLC analysis, and the molecular weight determined by mass spectrometry is 1682.90Da, which is consistent with the retention time and molecular weight of the purified polypeptide, and the fragments of the secondary mass spectrum are consistent with the purified polypeptide fragments.

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Abstract

The invention discloses a polypeptide for promoting tissue repair and an application of the polypeptide. The amino acid sequence of the polypeptide is Ile-Glu-Lys-Arg-Ile-Pro-Phe-Ser-His-Asp-Asp-Arg-Leu-Gly. According to the invention, a modern biochemical technical means is adopted; the polypeptide for promoting tissue repair in the drosophila melanogaster is subjected to tracking, screening andevaluating; and the steps of ethanol extraction, extraction, ion exchange resin, reversed-phase high performance liquid chromatography and other purification methods are carried out to obtain the product. The experimental results show that the polypeptide can promote proliferation and migration of human immortalized epidermal cells, is low in toxic and side effects, can be used for preparing medicines or skin care products for whitening, removing wrinkles, removing color spots and the like, can obviously improve the skin beauty, and has a good application prospect.

Description

technical field [0001] The invention relates to an active polypeptide, in particular to an active polypeptide isolated from a fruit fly extract and capable of promoting tissue repair, and belongs to the technical field of biomedicine. Background technique [0002] Skin wound healing refers to the healing process after the skin tissue is severed or damaged due to external force, including the repair and regeneration of various tissues, such as cell proliferation, differentiation, migration and other processes. Regeneration of the epidermis and other tissues usually occurs within 24 hours after trauma. The basal cells at the wound edge begin to proliferate and migrate to the center of the wound under the clot to form a single layer of epithelium covering the surface of the granulation tissue. Therefore, promoting tissue cell proliferation, migration, and repair is very important for wound healing. [0003] Peptide drugs have unique advantages in promoting wound healing, with ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K1/14C07K1/18C07K1/20A61K8/64A61Q19/02A61Q19/08A61K38/10A61P17/00A61P1/04
CPCC07K7/08A61K8/64A61Q19/02A61Q19/08A61P17/00A61P1/04A61K38/00
Inventor 刘睿段金廒
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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