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New Periplaneta Americana polypeptide capable of promoting tissue repair, and applications thereof

A technology for tissue repair and Periplaneta americana, which is applied in the fields of traditional Chinese medicine, natural medicine and daily chemicals, and can solve problems such as the application of peptides to promote tissue repair, unspecified effective components or active ingredients, and unclarified chemical composition of polypeptide parts. Achieve the effects of easy mass production, promotion of proliferation, and stable structure

Active Publication Date: 2017-07-25
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] It has been reported in the literature that the ethanol extract of Periplaneta americana can promote mucosal repair (Zhejiang Journal of Integrated Traditional Chinese and Western Medicine, 2012, 9: 689-691) and promote cell migration (Chinese Medicine and Clinic, 2013, 1: 34-36), but has not Clarify its effective components or active ingredients; Chinese invention patent ZL201210137422.4 discloses a preparation method of a cockroach polypeptide substance and its anti-herpes virus medical use, and Chinese invention patent ZL201210137410.1 introduces a preparation of a cockroach polypeptide and its Anti-Gram-positive and negative bacteria for medical purposes, but the above two patents not only did not clarify the specific chemical composition of the polypeptide part, but also did not involve the application of the polypeptide in promoting tissue repair

Method used

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  • New Periplaneta Americana polypeptide capable of promoting tissue repair, and applications thereof
  • New Periplaneta Americana polypeptide capable of promoting tissue repair, and applications thereof
  • New Periplaneta Americana polypeptide capable of promoting tissue repair, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Separation and structural identification of polypeptide Periplapeptide B

[0037] (1) 5 kg of Periplaneta americana medicinal materials, crushed into coarse powder, extracted 3 times by percolation with 80% (v / v) ethanol solution of 3 times the mass, each time for 24 hours; the combined extracts were concentrated under reduced pressure to a thick paste of about 1.5L, add 10 times the volume of hot water (70°C), keep it warm for 12 hours, separate layers, and discard the upper layer of oil; the lower layer of aqueous solution is concentrated and loaded on a reversed-phase C-18 chromatographic column, first with a volume percentage of 30 % methanol for 6 column volumes, and then eluted with 50% and 75% methanol aqueous solution for 6 column volumes respectively, followed by HPLC-MS analysis and detection, collected fractions mainly containing polypeptide compounds, combined fractions, Concentrate under reduced pressure or vacuum dry to obtain the total polypepti...

Embodiment 2

[0047] The synthesis of embodiment 2 new polypeptide Periplapeptide B

[0048] The new peptide Periplapeptide B was synthesized by using the conventional solid-phase synthesis method of an automatic peptide synthesizer, and through the processes of resin swelling, deprotection, washing, amino acid dissolution, amino acid activation, and condensation.

Embodiment 3

[0049] Example 3 Promoting Effect of New Polypeptide Periplapeptide B on Balb / c 3T3 Cell Proliferation

[0050] 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 3000 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. Discard the medium, add starvation medium containing 0.5% FBS for 24 hours to synchronize the cells; discard the starvation medium, add complete medium to configure different concentrations of peptide Periplapeptide B, 37°C, 5% CO 2 Incubator for 48h. Add CCK8 reagent at 10 μL / well, incubate at 37°C for 1 to 4 hours, and detect with dual wavelengths of 450nm and 630nm on a microplate reader. Calculate the cell survival rate and repeat the experiment at least 3 times;

[0051] Test results such as Figure 4 As show...

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PUM

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Abstract

The present invention relates to the field of traditional Chinese medicines, natural medicines and daily chemicals, particularly to a new Periplaneta Americana polypeptide capable of promoting tissue repair, and applications thereof, wherein the new polypeptide capable of promoting tissue repair is named Periplapeptide B, has a structure represented by a formula I, can promote the proliferation of mouse embryonic fibroblasts and the migration of human immortalized epidermal cells at a low concentration, has advantages of low toxic-side effect and good application prospect, and can be used for preparing drugs for treatment of trauma, scald and ulcer, or cosmetics for improving skin beauty, and daily chemicals. The formula I is H-Ala-Ala-Pro-Pro-Ser-Asn-Leu-Lys-Glu-Val-Pro-Ile-Ile-Ala-Tyr-OH.

Description

technical field [0001] The invention relates to the fields of traditional Chinese medicine, natural medicine and daily chemicals, in particular to a new Periplaneta americana polypeptide 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, peptides such as epidermal growth factor (hEGF) and basic fibroblast growth factor (bFGF) have obvious effects of repairing wounds, skin care, anti-wrinkle and anti-aging, but their high preparation cost and poor sta...

Claims

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Application Information

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IPC IPC(8): C07K7/08A61K8/64A61Q19/02A61Q19/08A61P1/04A61P1/02A61P17/02
CPCA61K8/64A61K38/00A61Q19/02A61Q19/08C07K7/08
Inventor 叶文才王磊刘忠范春林
Owner JINAN UNIVERSITY
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