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A kind of skin protection peptide hw-p1 and its preparation method and application

A HW-P1, skin protection technology, applied in the field of biomedicine, can solve problems such as synthesis difficulties, excessive wound repair, hypertrophic scars, etc., to achieve high efficiency and strong activity, and promote strong activity.

Active Publication Date: 2022-05-10
KUNMING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, these two types of drugs have certain limitations. The former is unstable, difficult to synthesize, and unsatisfactory in activity.
The latter requires harsh storage conditions, is prone to inactivation during transportation, storage, and use, and more importantly, it can cause excessive wound repair, leading to hypertrophic scars

Method used

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  • A kind of skin protection peptide hw-p1 and its preparation method and application
  • A kind of skin protection peptide hw-p1 and its preparation method and application
  • A kind of skin protection peptide hw-p1 and its preparation method and application

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preparation example Construction

[0019] The preparation method of the skin protection peptide HW-P1 of the present invention comprises the following steps:

[0020] A. Take the skin secretion of the tiger frog in Yunnan by electric stimulation, dissolve it in PBS, freeze-dry it in vacuum to obtain material a, and store it at -80°C for later use;

[0021] B. Dissolving material a in water to obtain material b, collecting the supernatant of material b after centrifugation to obtain material c, and obtaining material d through ultrafiltration of material c;

[0022] C. Dissolving material d in water to obtain material e, and performing high performance liquid chromatography reverse phase chromatography on material e to obtain the target skin protection peptide HW-P1.

[0023] The water described in the B step is ultrapure water.

[0024] The centrifugation described in the B step is to centrifuge at a rotating speed of 10000-15000r / min for 15-25min.

[0025] The temperature of the centrifugation is controlled ...

Embodiment 1

[0032] Isolation, purification and identification of a novel skin wound repairing peptide HW-P1

[0033] 1. Separation and purification

[0034] The skin secretions (dissolved in PBS) of tiger frogs collected from Yunnan were collected by electrical stimulation, vacuum freeze-dried, and stored at -80° for later use.

[0035] Step 1: Redissolve the lyophilized secretion in ultrapure water, then centrifuge at 12000×g for 20 minutes at 4°C, collect the supernatant, and then ultrafilter it with an Amicon ultrafilter to retain The molecular weight is 10 kDa (MerckMillipor, Germany).

[0036] The second step: high performance liquid chromatography reversed phase chromatography:

[0037] Take the sample dissolved in deionized water obtained in the first step, and load it onto a Hypersil ODS2 5 mm column (product of Yilite, with a size of 4.6 mm × 300 mm) equilibrated in advance with ultrapure water (containing 0.1% trifluoroacetic acid). ), the experimental instrument is Waters 15...

Embodiment 2

[0042] Detection of HaCaT Cell Scratch Repair Activity of Peptide HW-P1

[0043] Human immortalized keratinized epithelial cells were cultured in cell culture flasks with DMEM / F12 (BI, Israel) medium containing 10% fetal bovine serum and 1% double antibodies (penicillin, streptomycin, 100 U / mL) ( HaCaT). Use a 24-well plate for cell scratch experiments, inoculate 2.5×10 5 HaCaT cultured for about 12–24 h, when the cells in each well were full, scratched each well with a yellow 200 μL pipette tip (Axygen, USA), then discarded the medium containing dead cells in each well, and used Each well was washed twice with phosphate buffered saline (PBS), and finally 500 μL of empty medium containing different concentrations of samples (100 pM, 1 nM, 10 nM) without fetal bovine serum was added to each well. Use a microscope (Zeiss, Germany) to take pictures every 12 hours to record the scratch healing status, and record continuously for 24 hours. We evaluated the percentage of cellular...

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Abstract

The invention discloses a skin protection peptide HW‑P1, its preparation method and application. The amino acid sequence of the skin protection peptide HW‑P1 is QNSYADLWCQFHYMC. The invention extracts an active polypeptide for promoting skin wound repair from the skin secretion of the tiger frog, and the polypeptide has the characteristics of natural source, high activity and the like. The purified peptide HW-P1 for promoting skin wound repair shows strong wound recovery activity, indicating that the "skin protection peptide HW-P1" of the present invention has great medical and cosmetic application prospects. The skin protection peptide HW‑P1 described in the present invention has the beneficial feature of highly effective and strong activity in promoting skin wound repair.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a skin protection peptide HW-P1 and its preparation method and application. Background technique [0002] The skin covers the surface of the human body, accounting for about 16% of the body weight, and its area can reach 1.2-2.0 square meters. It is one of the largest organs in the human body. The skin is in direct contact with the outside world and can block the invasion of foreign bodies and pathogenic microorganisms. It has an important protective effect on the human body and is the first line of defense for human immunity; at the same time, it can prevent the loss of tissue fluid. There are rich sensory nerve endings in the skin, which can feel a variety of stimuli. In addition, the skin also plays a very important role in maintaining a constant body temperature. However, in daily life, the skin is extremely vulnerable to damage caused by external factors. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08C07K1/34C07K1/16C07K1/36A23L33/18A61K38/10A61P17/00A61K8/64A61Q19/00
CPCC07K7/08A23L33/18A61P17/00A61K8/64A61Q19/00A23V2002/00A61K38/00A23V2200/318A23V2250/55
Inventor 杨新旺王滢唐璟冯卓杨莹尹赛格刘乃心孙慧玲李依林傅阳
Owner KUNMING MEDICAL UNIVERSITY
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