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Novel polypeptide and Periplaneta americana total polypeptide extract capable of promoting tissue repair, and application thereof

A tissue repair and American cockroach technology, applied in the field of traditional Chinese medicine and natural medicine, can solve the problems of unclear effective components or active ingredients, not involved in promoting tissue repair, and unclear chemical composition types, etc., to improve skin beauty and stability. Good sex and wrinkle removal effect

Active Publication Date: 2017-05-24
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 active components or active ingredients; Chinese invention patent 200710067707.4 introduces the method of preparing anti-tumor active parts of Periplaneta americana with reversed-phase materials, but does not clarify the type of chemical components and does not involve the role of promoting tissue repair; Chinese invention patent ZL201210137422. 4 discloses a preparation method of a cockroach polypeptide substance and its anti-herpes virus medical use. Chinese invention patent ZL201210137410.1 introduces a preparation of a cockroach polypeptide and its anti-Gram-positive and Gram-negative bacteria. However, the above-mentioned patent Not only did not clarify the specific chemical composition of the polypeptide part, but also did not involve the application of the total polypeptide and polypeptide monomers in promoting tissue repair

Method used

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  • Novel polypeptide and Periplaneta americana total polypeptide extract capable of promoting tissue repair, and application thereof
  • Novel polypeptide and Periplaneta americana total polypeptide extract capable of promoting tissue repair, and application thereof
  • Novel polypeptide and Periplaneta americana total polypeptide extract capable of promoting tissue repair, and application thereof

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Effect test

Embodiment 1

[0044] The preparation of embodiment 1 Periplaneta americana total polypeptide extract

[0045]Periplaneta americana medicinal material 5kg, pulverized into coarse powder, extracted 3 times by percolation with 3 times the mass of 80% (v / v) ethanol, 24 hours each time. Combine the extracts, concentrate 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 the layers, and discard the upper layer of oil. The lower aqueous solution was concentrated and loaded on a reversed-phase C-18 chromatographic column. First, 6 column volumes were eluted with 30% methanol by volume percentage, and then 6 column volumes were eluted with 50% and 75% methanol aqueous solutions by volume percentage respectively. Use HPLC-MS to analyze and track detection, collect the fractions that mainly contain polypeptide compounds, combine fractions, concentrate under reduced pressure or dry in vacuo, obtain the total polypeptid...

Embodiment 2

[0046] Example 2 Separation and structural identification of polypeptide Periplapeptide A

[0047] (1) Get 20g of Periplaneta americana total polypeptide extract and carry out reverse-phase silica gel (ODS) column chromatography, with methanol-water as eluent, according to methanol and water volume ratio is 10:90, 20:80, 30:70 , 50:50, 70:30, 90:10 elution gradient to obtain 6 main components Fr.A~Fr.F;

[0048] (2) Load the fraction Fr.C (2.5g) obtained in step (1) onto a Sephadex G-50 chromatographic column, use water as the mobile phase, and elute at a flow rate of 0.5mL / min. After HPLC-MS Analyze and detect, combine and collect similar fractions to obtain 5 sub-fractions Fr.C1~Fr.C5;

[0049] (3) Concentrate the Fr.C4 obtained in step (2) and dissolve it in methanol with a volume fraction of 5%, then separate and purify it by reverse-phase preparative HPLC, using acetonitrile-water with a volume ratio of 40:60 as the eluent, the flow rate Elution was carried out at 3 mL / ...

Embodiment 3

[0056] The synthesis of embodiment 3 new polypeptide Periplapeptide A

[0057] The new polypeptide Periplapeptide A was synthesized by using the conventional solid-phase synthesis method of an automatic peptide synthesizer, through resin swelling, deprotection, washing, amino acid dissolution, amino acid activation, and condensation, and 5.0 g of the product was obtained.

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Abstract

The invention specifically relates to novel polypeptide and Periplaneta americana total polypeptide extract capable of promoting tissue repair, and application thereof, belonging to the fields of traditional Chinese medicines and crude drugs. The novel polypeptide capable of promoting tissue repair is named as Periplapeptide A and has a structure as shown in a formula I H-Ala-Ala-Pro-Tyr-Ala-His-Val-Ala-Ala-Pro-Tyr-Ala-His-Phe-OH. The Periplaneta americana total polypeptide extract capable of promoting tissue repair is prepared by subjecting the bodies of Periplaneta americana to alcohol extraction, degreasing and enrichment via a reversed-phase material, wherein the mass percentage content of total polypeptide is greater than 50%, and the extract contains Periplapeptide A. The novel polypeptide and the Periplaneta americana total polypeptide extract containing novel polypeptide in the invention can promote propagation of mouse embryo fibroblasts and migration of immortalized human epidermal cells at low concentrations, have low toxic and side effect and good medicinal prospects, and are applicable to preparation of medicines used for treating wounds, burns and ulcers or cosmetics and daily chemical articles used for beautifying the skin.

Description

technical field [0001] The invention relates to the field of traditional Chinese medicine and natural medicine, in particular to a new polypeptide for promoting tissue repair, an extract of total polypeptide of Periplaneta americana 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, which are very harmful. 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 preparati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61K35/64A61K8/98A61P17/00A61P17/02A61Q19/00
Inventor 叶文才王磊刘忠范春林蒋文贤高贤达
Owner JINAN UNIVERSITY
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