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Periplaneta americana tissue repair factor PA1 and application thereof

A technology of tissue repair and Periplaneta americana, which is applied to the peptide "tissue repair factor PA1" that promotes tissue repair and its application field, which can solve the problems of active ingredients that need to be studied in depth, and achieve a product that is easy to prepare in large quantities, promotes migration, and removes wrinkles Effect

Active Publication Date: 2020-08-04
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The drug Kangfuxin liquid made from Periplaneta americana has good curative effect in treating wounds, scalds and peptic ulcers, but its active ingredients need to be further studied

Method used

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  • Periplaneta americana tissue repair factor PA1 and application thereof
  • Periplaneta americana tissue repair factor PA1 and application thereof
  • Periplaneta americana tissue repair factor PA1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Isolation and structure identification of tissue repair factor PA1

[0043] (1) Periplaneta americana medicinal material 1kg, crushed into coarse powder, extracted by percolation with 3 times the mass of 80% (v / v) ethanol solution 3 times, each time for 24 hours; combined extracts, concentrated under reduced pressure to thick paste 200 ~300mL, add 10~15 times the volume of hot water (70°C), keep it warm for 12 hours, separate layers, and discard the upper layer of oil. After the aqueous solution was concentrated, it was loaded on a reversed-phase C-18 chromatographic column, and 6 column volumes were first eluted with 30% methanol by volume, and then 6 column volumes were eluted with 50% and 75% methanol aqueous solutions by volume percentage, and then 6 column volumes were eluted with HPLC-MS analysis followed detection, collected the fractions mainly containing polypeptide compounds, combined the fractions, concentrated under reduced pressure or vacuum dried...

Embodiment 2

[0052] Example 2 Solid phase synthesis of tissue repair factor PA1

[0053] The conventional solid-phase synthesis method of peptide synthesizer is used to complete the synthesis of tissue repair factor PA1 through the processes of resin swelling, deprotection, washing, amino acid dissolution, amino acid activation, condensation, and cutting. The method of liquid phase-mass spectrometry was used to compare the synthesized and isolated tissue repair factor PA1 samples, and the results showed that their retention times and molecular weights were consistent ( Figure 8 ).

Embodiment 3

[0054] Example 3 Promoting effect of tissue repair factor PA1 on the proliferation of Balb / c 3T3 cells

[0055] Cells in the logarithmic growth phase (Balb / c 3T3 cell line, purchased from the American Type Culture Collection, ATCC) were digested with trypsin, centrifuged, and resuspended for cell counting, counted as 10,000 cells Inoculate each well in a 96-well plate, tap until the cells are evenly dispersed, and block with PBS. 37°C, 5% CO 2 Incubate overnight. After the cells adhered to the wall, the original medium was discarded, and then DMEM medium (containing 10% fetal bovine serum) containing different concentrations of "tissue repair factor PA1" was added, at 37°C, 5% CO 2 Incubate in the incubator for 24h. Add 30 μL of MTT solution to each well, continue to incubate for 4 h, discard the upper layer of MTT, add 100 μL of DMSO to each well, shake for 10 min to fully dissolve the crystals, and measure the OD value of each well at 570 nm with an enzyme-linked immunoso...

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Abstract

The invention relates to the field of medicines and daily chemicals, in particular to a periplaneta americana new polypeptide for promoting tissue repair and application of the periplaneta americana new polypeptide, the structure of the periplaneta americana new polypeptide is shown as a formula I, and the periplaneta americana new polypeptide is named as a tissue repair factor PA1. The new polypeptide can promote proliferation of mouse embryo fibroblasts and migration of human immortalized epidermal cells even at a low concentration, has low toxic and side effects and has a good application prospect, so that the polypeptide can be used for preparing medicines for treating wounds, scalds and ulcers or cosmetics and daily chemical products for improving skin beauty.

Description

technical field [0001] The invention relates to the fields of medicine and daily chemicals, in particular to a polypeptide "tissue repair factor PA1" for promoting tissue repair and its application. Background technique [0002] The healing process after tissue trauma is a process of proliferation, differentiation, migration and apoptosis of various repair cells. Acute and chronic injuries such as inflammation, ulcers, burns, trauma, surgery, and congenital deformities can cause large areas of skin defects, refractory ulcers, and scars, and trigger many local and systemic reactions, causing serious harm. Peptide drugs have unique advantages in promoting wound healing, preventing scars and treating chronic ulcers. Among them, polypeptides such as epidermal growth factor (hEGF) and basic fibroblast growth factor (bFGF) have obvious functions of repairing wounds, skin care, Anti-wrinkle and anti-aging effects , but their high preparation cost and poor stability limit their ap...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435A61K38/17A61P17/02A61P17/00A61K8/64A61Q19/00A61Q19/08
CPCA61K8/64A61K38/00A61Q19/00A61Q19/08A61P17/00A61P17/02C07K14/43563
Inventor 叶文才王磊范春林曹佳青
Owner JINAN UNIVERSITY
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