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Skin protection peptide OM-TV16, and preparation method and application thereof

An OM-TV16, skin protection technology, applied in the field of biomedicine, can solve problems such as insufficient attention, and achieve the effect of large medical application prospects, high efficiency and strong activity, and the effect of promoting strong activity

Active Publication Date: 2020-01-03
杨莹
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Generally speaking, highly active, stable, and low-cost skin wound repair active peptides have broad market prospects, but the current research and development of peptides have not received enough attention, so exploring new wound repair peptides as wound repair drugs lead molecules are of great importance

Method used

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  • Skin protection peptide OM-TV16, and preparation method and application thereof
  • Skin protection peptide OM-TV16, and preparation method and application thereof
  • Skin protection peptide OM-TV16, and preparation method and application thereof

Examples

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preparation example Construction

[0017] The preparation method of the skin protection peptide OM-TV16 of the present invention comprises the following steps:

[0018] A. Take the skin secretion of Guizhou green stink frog by electric stimulation method, dissolve it in PBS, and freeze-dry it in vacuum to obtain material a, and store it at -80°C for later use;

[0019] 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;

[0020] 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 OM-TV16.

[0021] The water described in the B step is ultrapure water.

[0022] The centrifugation described in the B step is to centrifuge at a rotating speed of 10000-15000r / min for 15-25min.

[0023] The temperature of the centrifugation...

Embodiment 1

[0030] Isolation, purification and identification of a novel skin wound repairing peptide OM-TV16

[0031] 1. Separation and purification

[0032] The skin secretions (dissolved in PBS) of green stinky frogs collected from Guizhou were collected by electrical stimulation, vacuum freeze-dried, and stored at -80°C for future use.

[0033] 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 with a molecular weight cut-off of 10 kDa (Merck Millipor, Germany).

[0034] The second step: high performance liquid chromatography reversed phase chromatography:

[0035] Take the sample dissolved in deionized water obtained in the first step, and load it on 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 ...

Embodiment 2

[0040] Detection of scratch repair activity of OM-TV16 in HaCaT cells

[0041] Human keratinocytes (HaCaT) were cultured in DMEM / F12 medium (BI, Israel) containing 10% fetal bovine serum (FBS, BI, Israel) and incubated at 37°C in an incubator containing 5% CO2 . Cells were then digested and transferred to 24-well plates (2.5 × 10 5cells / well) and culture HaCaT for 12-24 hours, after the monolayer of cells covered the bottom of the well, use a sterile yellow 200 μL pipette tip (Axygen, USA) to scratch vertically in the center of the bottom of the well, and wash twice with PBS Remove the scratched cells. Subsequently, 500 µL of DMEM / F12 serum-free medium containing 25 nM, 50 nM, 100 nM of OM-TV16 and 50 nM of OM-LV20 (positive control) was added to each well. Add 10 mL PBS to the serum-free medium, three wells in each group. Images of scratch-healing cell monolayers were taken with an inverted microscope (Primovert microscope, Zeiss, Germany) at 0, 6, 12, 18, and 24 hours af...

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Abstract

The invention discloses a skin protection peptide OM-TV16, and a preparation method and application thereof. The amino acid sequence of the skin protection peptide OM-TV16 is TVWGFRPTNPKPKPPG. The invention extracts an active peptide capable of accelerating skin wounds to be repaired from the skin excretions of odorrana margaretae, and the peptide has the characteristics of natural sources, high activity and the like. The peptide OM-TV16 which is obtained by purification and is capable of accelerating the skin wounds to be repaired has a high wound recovery activity so as to show that the ''skin protection peptide OM-TV16'' has a wide medical care and cosmetics application prospect. The skin protection peptide OM-TV16 disclosed by the invention has the beneficial characteristics of a highactivity for efficiently accelerating skin wounds to be repaired.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a skin protection peptide OM-TV16 and its preparation method and application. Background technique [0002] As the first physical barrier to protect various tissues and organs in the body, the skin wraps around the surface of the human body and plays an important role in maintaining the body's water and electrolyte balance, regulating body temperature, feeling pain, temperature, touch and pressure, and protecting the body from external environmental influences. However, because the skin is directly exposed to the external environment without the protection of fur scales, it may cause skin damage under the influence of factors such as mechanical external force, burns or chronic diseases, thereby affecting the normal function of the skin. Although the body can repair wounds by itself, if the repair process goes wrong, it may make it difficult for the damaged skin to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K1/36C07K1/34C07K1/20A61K38/10A61K8/64A61P17/02A61P1/02A61Q19/00A23L33/18
CPCC07K7/08A61K8/64A61P17/02A61P1/02A61Q19/00A23L33/18A61K38/00
Inventor 杨莹杨新旺李奕平赵建唐璟陶剑
Owner 杨莹
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