CN111269292A - Housefly polypeptide with function of promoting tissue repair and preparation method and application thereof - Google Patents
Housefly polypeptide with function of promoting tissue repair and preparation method and application thereof Download PDFInfo
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- CN111269292A CN111269292A CN202010142284.3A CN202010142284A CN111269292A CN 111269292 A CN111269292 A CN 111269292A CN 202010142284 A CN202010142284 A CN 202010142284A CN 111269292 A CN111269292 A CN 111269292A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Abstract
The invention discloses a housefly polypeptide with the function of promoting tissue repair, a preparation method and an application thereof, wherein the amino acid sequence of the polypeptide is as follows: Leu-Ala-Asp-Glu-Glu-Ser-Arg-Glu-Arg-Ala-Thr-Leu-Leu-Gly. The polypeptide for promoting tissue repair in the housefly is subjected to tracking screening and evaluation by adopting modern biochemical technical means, and is prepared by purifying methods such as ethanol extraction, ion exchange resin, reversed-phase high performance liquid chromatography and the like.
Description
Technical Field
The invention relates to an active polypeptide, in particular to an active polypeptide which is separated from a housefly extract and has the effect of promoting tissue repair and a preparation method thereof, belonging to the technical field of biological medicines.
Background
The healing process of skin wound refers to the healing process of skin tissue after being separated or defected due to the action of external force, and includes the repair and regeneration of various tissues, such as cell proliferation, differentiation, migration and the like. Epidermal and other tissue regeneration usually occurs within 24 hours after wounding, and the basal cells at the wound edges begin to proliferate and migrate under the clot towards the center of the wound, forming a monolayer of epithelium covering the surface of granulation tissue. Therefore, promotion of tissue cell proliferation, migration, and repair is very important for wound healing.
The polypeptide medicine has unique advantages in promoting wound healing, and has strong effect and high safety. Polypeptides such as epidermal growth factor, basic fibroblast growth factor and the like have obvious effect of repairing wounds; modern researches find that the active polypeptide from animal sources also has very good effect of promoting wound repair.
Ben Cao gang mu relates to jin Shi Hu, which can be used to treat infantile malnutrition, fever, delirium, dysentery with toxic pathogen and vomiting. The chrysomyia larvae has the effects of strengthening spleen, removing food retention, clearing heat and removing malnutrition. The previous research of the applicant finds that the housefly extract has a good effect of promoting the proliferation and migration of human immortalized epidermal cells (HaCat), but the effective components of the housefly extract are still unclear.
Disclosure of Invention
The purpose of the invention is as follows: on the basis of earlier-stage research, the housefly polypeptide for promoting tissue repair is prepared by carrying out tracking evaluation on components for promoting tissue repair in a housefly extract through a large number of experimental screens and adopting modern biochemical technical means and purifying methods such as extraction, strong oxygen ion exchange chromatography, reversed-phase high-performance liquid chromatography and the like.
The technical scheme is as follows: in order to achieve the above purpose, the invention adopts the technical scheme that:
a housefly polypeptide that promotes tissue repair, the polypeptide having the amino acid sequence: Leu-Ala-Asp-Glu-Glu-Ser-Arg-Glu-Arg-Ala-Thr-Leu-Leu-Gly.
The preparation method of the housefly polypeptide for promoting tissue repair comprises the following steps:
(1) preparation of housefly extract: extracting pulverized or non-pulverized whole Musca domestica with ethanol, and concentrating to obtain Musca domestica extract;
(2) and (3) extraction: dispersing the housefly extract obtained in the step (1) by using a methanol solution, adding chloroform, sufficiently shaking, standing, removing a chloroform layer, centrifuging a methanol solution part, concentrating and drying to obtain a methanol part of the housefly extract;
(3) strong oxygen ion exchange resin separation: re-dissolving the methanol part obtained in the step (2) with an ammonium formate solution, repeatedly loading a strong oxygen ion exchange resin to separate the small column, washing the small column with 5-10 mM of the ammonium formate solution for 7-8 times, then eluting the small column with the ammonium formate solution, and collecting eluent to obtain a housefly polypeptide part;
(4) and (3) separating and purifying by reverse phase chromatography: re-dissolving the housefly polypeptide part obtained in the step (3) with a formic acid solution, and performing reversed-phase preparative high performance liquid chromatography separation, wherein the mobile phase comprises the following steps: the phase A is formic acid with the volume concentration of 0.1 percent; phase B is methanol, and gradient elution is carried out; flow rate: 5-10 mL/min; column temperature: at 30 ℃, the detection wavelength is 220nm, and the housefly polypeptide is obtained by separation and collection, and the amino acid sequence of the polypeptide is as follows: Leu-Ala-Asp-Glu-Glu-Ser-Arg-Glu-Arg-Ala-Thr-Leu-Leu-Gly.
Preferably, the method for preparing housefly polypeptide for promoting tissue repair comprises the following steps:
(1) preparation of housefly extract: extracting pulverized or pulverized whole body of Musca domestica with 70 vol% ethanol, filtering to obtain extract, and concentrating;
(2) and (3) extraction: dispersing the housefly extract obtained in the step (1) by using a methanol solution with the volume concentration of 70%, adding 1/10 volumes of chloroform, fully shaking, standing, removing a chloroform layer, centrifuging a methanol solution part, concentrating and drying to obtain a methanol extract part of the housefly;
(3) strong oxygen ion exchange resin separation: redissolving the methanol extract part of the housefly in 10mM ammonium formate solution, wherein the ammonium formate solution contains 25% acetonitrile and has the pH value of 2.7-3; repeatedly loading the strong oxygen ion exchange resin small column to ensure that the sample is fully absorbed, washing the strong oxygen ion exchange resin small column for 7-8 times by using 10mM ammonium formate solution containing 25% acetonitrile and pH 2.7-3, eluting the strong oxygen ion exchange resin small column by using 0.4M ammonium formate solution containing 25% acetonitrile and pH 5, and collecting eluent to obtain housefly polypeptide parts;
(4) and (3) separating and purifying by reverse phase chromatography: re-dissolving the housefly polypeptide part obtained in the step (3) by using a formic acid solution with the volume concentration of 0.1%, and performing reversed-phase preparative high performance liquid chromatography separation, wherein the mobile phase comprises the following components in parts by weight: the phase A is formic acid with the volume concentration of 0.1 percent; phase B is methanol, and gradient elution is carried out; flow rate: 10 mL/min; column temperature: at 30 ℃, the detection wavelength is 220nm, and the housefly polypeptide is obtained by separation and collection, and the amino acid sequence of the polypeptide is as follows: Leu-Ala-Asp-Glu-Glu-Ser-Arg-Glu-Arg-Ala-Thr-Leu-Leu-Gly.
Preferably, in the preparation method of the housefly polypeptide for promoting tissue repair, the gradient elution mode is as follows: and (3) the concentration of the mobile phase B is increased to 35% by volume from 2% by volume in a gradient manner at a flow rate of 10mL/min for 0-15 min.
The housefly polypeptide for promoting tissue repair is applied to preparation of medicines, cosmetics or daily chemical products for promoting tissue repair or improving skin aesthetic feeling. The medicine is in the form of tablet, capsule, injection, granule, gel ointment, liniment, etc.
The cosmetic or daily chemical product is in the form of ointment, liniment, patch (such as facial mask), cream (such as facial cream), detergent (such as soap, bath lotion, etc.), emulsion, gel or toner, etc.
The invention has the following advantages and effects for the prior art:
(1) the invention firstly separates, purifies and identifies the polypeptide with the function of promoting tissue repair from the housefly, and the amino acid sequence is as follows: Leu-Ala-Asp-Glu-Glu-Ser-Arg-Glu-Arg-Ala-Thr-Leu-Leu-Gly.
(2) The polypeptide of the invention can promote the proliferation and migration of HaCat under a lower concentration (5 mu M).
(3) The polypeptide of the invention has good stability, does not hydrolyze under the action of gastrointestinal tract, and can be used for tissue repair and digestive tract ulcer.
(4) The polypeptide of the invention has the functions of whitening, removing wrinkles, removing color spots and the like, and can obviously improve the aesthetic feeling of skin.
Drawings
FIG. 1 is a mass spectrum of a polypeptide provided by the present invention.
FIG. 2 shows the proliferation of HaCat cells by housefly repair peptides.
FIG. 3 is a line graph showing the migration promotion of HaCat cells by housefly repair peptides.
Detailed Description
The present invention is further illustrated by the following examples, which are intended to be purely exemplary and are not intended to limit the scope of the invention, as various equivalent modifications of the invention will occur to those skilled in the art upon reading the present disclosure and fall within the scope of the appended claims.
Example 1 a method for preparing housefly repair peptides, comprising the steps of:
(1) preparation of housefly extract: extracting pulverized or pulverized whole body of Musca domestica with 70% ethanol, and concentrating;
(2) and (3) extraction: dispersing the housefly extract concentrate with 70% methanol solution, adding about 1/10 volumes of chloroform, shaking thoroughly, standing, discarding chloroform layer, centrifuging the methanol solution part, concentrating, and drying to obtain housefly extract methanol part;
(3) strong oxygen ion exchange resin (SCX) separation: re-dissolving the musca domestica extract methanol part obtained in the step (2) with 10mM ammonium formate solution (containing 25% acetonitrile and having a pH of 2.7-3), repeatedly loading the SCX cartridge to ensure that the sample is fully adsorbed, washing the SCX cartridge for 7-8 times with 10mM ammonium formate solution (containing 25% acetonitrile and having a pH of 2.7-3), eluting the SCX cartridge with 0.4M ammonium formate solution (containing 25% acetonitrile and having a pH of 5), and collecting the eluent to obtain musca domestica polypeptide part;
(4) and (3) separating and purifying by reverse phase chromatography: re-dissolving the housefly polypeptide part obtained in the step (3) with 0.1% formic acid solution, and performing reversed-phase preparative high performance liquid chromatography separation, wherein the mobile phase comprises the following components in parts by weight: the phase A is formic acid with the volume concentration of 0.1 percent; phase B is methanol, and gradient elution is carried out; flow rate: 10 mL/min; column temperature: at 30 ℃, the detection wavelength is 220nm, and the housefly polypeptide is obtained by separation and collection, and the amino acid sequence of the polypeptide is as follows: Leu-Ala-Asp-Glu-Glu-Ser-Arg-Glu-Arg-Ala-Thr-Leu-Leu-Gly.
Example 2 sequence analysis of housefly repair peptides, comprising the following steps:
the housefly repairing peptide prepared in the example 1 is taken, nano-LC-ESI-MS/MS is adopted to analyze the amino acid sequence of the polypeptide, and the mass spectrum condition is as follows: ESI source, scan mode: in a positive ion mode, the primary full-scanning range of the mass spectrum is 300-2000 m/z, and the separation width is 3 Da; the tandem mass spectrometry adopts a secondary mass spectrometry scanning mode depending on primary mass spectrometry data, and 5 ions with the highest ion intensity in the primary mass spectrometry are sequentially selected for carrying out Collision Induced Dissociation (CID) secondary tandem mass spectrometry. The amino acid sequence of the housefly repairing peptide determined according to the obtained MS/MS map is as follows: Leu-Ala-Asp-Glu-Glu-Ser-Arg-Glu-Arg-Ala-Thr-Leu-Leu-Gly as shown in FIG. 1.
Example 3 Synthesis of Musca domestica repair peptides
According to the amino acid sequence obtained in example 2, the dipeptidyl peptidase-4 inhibiting peptide Leu-Ala-Asp-Glu-Glu-Ser-Arg-Glu-Arg-Ala-Thr-Leu-Leu-Gly was synthesized by solid phase synthesis, the synthesized polypeptide had a purity of 98% by HPLC analysis, a molecular weight of 1558.79Da by mass spectrometry, and the fragment of the second mass spectrum was consistent with the fragment of the purified polypeptide.
Example 4 Musca domestica repair peptide promotes HaCat cell proliferation
HaCat seed plate: the cells were digested with 0.1% trypsin, gently tapped, the culture medium was added to stop the trypsin action, the cells were carefully collected in a centrifuge tube, centrifuged (1000rpm, 5min), the supernatant was removed, the cells were resuspended in 10ml of 10% FBS-DMEM medium and counted. Adjusting the cell suspension concentration to 2X 103cells/well, addInto 96-well plates (marginal wells filled with sterile PBS). 5% CO2Incubate at 37 ℃ and adhere overnight.
Administration: removing supernatant, adding housefly repairing peptide solution with final concentration of 20, 10, 5 μ g/ml prepared with 10% FBS-DMEM medium, each well having 200 μ l, each concentration having 3 multiple wells, blank well having 200 μ l 10% FBS-DMEM medium, and administering 5% CO2Incubate at 37 ℃ for 24 hours. The supernatant was removed from each well, washed once with 200. mu.l sterile PBS, 150. mu.l culture medium was added, 10. mu.l cck-8 assay solution was added, blank control wells were set, wells contained no cells, 150. mu.l culture medium and 10. mu.l lcck-8 assay solution were added. After incubation for 3 hours in an incubator at 37 ℃, the absorbance of each well was measured at 450nm, and the cell viability was calculated.
As shown in FIG. 2, the housefly repair peptides showed significant proliferation effect on HaCat at 5, 10, 20. mu.g/ml concentration (p < 0.05; p <0.01) compared to the blank group.
Example 5 Musca domestica repair peptides promoting HaCat cell migration
HaCat seed plate: the cells were digested with 0.1% trypsin, gently tapped, the culture medium was added to stop the trypsin action, the cells were carefully collected in a centrifuge tube, centrifuged (1000rpm, 5min), the supernatant was removed, the cells were resuspended in 10ml of 10% FBS-DMEM medium and counted. Adjusting the cell suspension concentration to 1X 105cells/well, add to 12-well plate. 5% CO2Incubate at 37 ℃ and adhere overnight. After the cells are fused, scratching damage is carried out on the cells in the holes by using the gun head, and scratches with the width of about 1mm are uniform and consistent. The vicinity of the scratch was gently tapped 3 times with serum-free DMEM medium to remove the scraped cells.
Administration: the housefly repairing peptide is prepared into solutions with the concentration of 20 mu g/ml by using 5% FBS-DMEM culture medium, 1ml of housefly repairing peptide solution is added into each well, each sample is provided with 3 multiple wells, and the blank well is 1ml of 5% FBS-DMEM culture medium. 5% CO after administration2Incubation at 37 ℃ and microscopic observation of cell fusion at 0, 24 and 36 hours, respectively.
The scratch width was measured by selecting 6 equidistant points on the graph at each time point in the graph using Image Pro Plus software, and the mobility was calculated as follows:
mobility ═ [ (initial width) - (width after incubation) ]/initial width × 100%
The experimental result is shown in fig. 3, a line graph is drawn according to the mobility, the abscissa is time, the ordinate is mobility, the housefly repairing peptide can remarkably promote healing of the HaCat scratch after being incubated for 24h (p is less than 0.05), and the housefly repairing peptide can remarkably promote healing of the HaCat scratch after being incubated for 36h (p is less than 0.01).
Example 5 the repair of Musca domestica repair peptides on skin lesions in mice
Selecting 40 male Kunming mice, 18-20g, raising in cages, freely taking food, and fasting for 12h before experiment. The mouse is anesthetized by intraperitoneal injection of 0.2ml of 1% sodium pentobarbital, the back hair is cut off, square wounds of 0.5cm multiplied by 0.5cm are symmetrically formed in the upper part and the lower part of the spinal epidermal skin, the whole layer of skin is cut off, and the wounds stop bleeding for later use. The next day, the mice were anesthetized with ether, and the wound was instilled with housefly repairing peptide and physiological saline 0.1ml at different concentrations, and changed once a day. Wound healing was observed on days 3, 5, and 10.
The wound healing rate was calculated as follows:
the wound healing rate is [ (original wound area) - (non-healing wound area) ]/original wound area × 100%; the results are shown in table 1, and show that the housefly repair peptide can remarkably promote the healing of the skin wound of the mouse (p < 0.01).
TABLE 1 statistical results of wound healing in mice
Significant differences with p <0.05 compared to model group; there were very significant differences with p < 0.01.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Sequence listing
<110> Nanjing university of traditional Chinese medicine
<120> housefly polypeptide with tissue repair promoting function, preparation method and application thereof
<141>2020-03-04
<160>1
<170>SIPOSequenceListing 1.0
<210>1
<211>14
<212>PRT
<213> Artificial sequence (2 Ambystoma latex x Ambystoma jeffersonia)
<400>1
Leu Ala Asp Glu Glu Ser Arg Glu Arg Ala Thr Leu Leu Gly
1 5 10
Claims (7)
1. A housefly polypeptide that promotes tissue repair, the polypeptide having an amino acid sequence that:
Leu-Ala-Asp-Glu-Glu-Ser-Arg-Glu-Arg-Ala-Thr-Leu-Leu-Gly。
2. the method for preparing housefly polypeptide for promoting tissue repair according to claim 1, comprising the steps of:
(1) preparation of housefly extract: extracting pulverized or non-pulverized whole Musca domestica with ethanol, and concentrating to obtain Musca domestica extract;
(2) and (3) extraction: dispersing the housefly extract obtained in the step (1) by using a methanol solution, adding chloroform, sufficiently shaking, standing, removing a chloroform layer, centrifuging a methanol solution part, concentrating and drying to obtain a methanol part of the housefly extract;
(3) strong oxygen ion exchange resin separation: re-dissolving the methanol part obtained in the step (2) with an ammonium formate solution, repeatedly loading a strong oxygen ion exchange resin to separate the small column, washing the small column with 5-10 mM of the ammonium formate solution, then eluting the small column with the ammonium formate solution, and collecting eluent to obtain a housefly polypeptide part;
(4) and (3) separating and purifying by reverse phase chromatography: re-dissolving the housefly polypeptide part obtained in the step (3) with a formic acid solution, and performing reversed-phase preparative high performance liquid chromatography separation, wherein the mobile phase comprises the following steps: the phase A is formic acid with the volume concentration of 0.1 percent; phase B is methanol, and gradient elution is carried out; flow rate: 5-10 mL/min; column temperature: at 30 ℃, the detection wavelength is 220nm, and the housefly polypeptide is obtained by separation and collection, and the amino acid sequence of the polypeptide is as follows: Leu-Ala-Asp-Glu-Glu-Ser-Arg-Glu-Arg-Ala-Thr-Leu-Leu-Gly.
3. The method for preparing housefly polypeptide for promoting tissue repair according to claim 2, comprising the steps of:
(1) preparation of housefly extract: extracting pulverized or pulverized whole body of Musca domestica with 70 vol% ethanol, filtering to obtain extract, and concentrating;
(2) and (3) extraction: dispersing the housefly extract obtained in the step (1) by using a methanol solution with the volume concentration of 70%, adding 1/10 volumes of chloroform, fully shaking, standing, removing a chloroform layer, centrifuging a methanol solution part, concentrating and drying to obtain a methanol extract part of the housefly;
(3) strong oxygen ion exchange resin separation: redissolving the methanol extract part of the housefly in 10mM ammonium formate solution, wherein the ammonium formate solution contains 25% acetonitrile and has the pH value of 2.7-3; repeatedly loading the strong oxygen ion exchange resin small column to ensure that the sample is fully absorbed, washing the strong oxygen ion exchange resin small column for 7-8 times by using 10mM ammonium formate solution containing 25% acetonitrile and pH 2.7-3, eluting the strong oxygen ion exchange resin small column by using 0.4M ammonium formate solution containing 25% acetonitrile and pH 5, and collecting eluent to obtain housefly polypeptide parts;
(4) and (3) separating and purifying by reverse phase chromatography: re-dissolving the housefly polypeptide part obtained in the step (3) by using a formic acid solution with the volume concentration of 0.1%, and performing reversed-phase preparative high performance liquid chromatography separation, wherein the mobile phase comprises the following components in parts by weight: the phase A is formic acid with the volume concentration of 0.1 percent; phase B is methanol, and gradient elution is carried out; flow rate: 10 mL/min; column temperature: at 30 ℃, the detection wavelength is 220nm, and the housefly polypeptide is obtained by separation and collection, and the amino acid sequence of the polypeptide is as follows: Leu-Ala-Asp-Glu-Glu-Ser-Arg-Glu-Arg-Ala-Thr-Leu-Leu-Gly.
4. The method for preparing housefly polypeptide for promoting tissue repair according to claim 2 or 3, wherein the gradient elution mode is as follows: and in 0-15 min, the concentration of the mobile phase B is increased from 2% to 35% in volume ratio in a gradient manner, and the flow rate is 10 mL/min.
5. Use of the housefly polypeptide for promoting tissue repair according to claim 1 in the preparation of a medicament, cosmetic or daily chemical for promoting tissue repair or improving skin aesthetic effect.
6. Use according to claim 5, characterized in that: the medicine is in the form of tablet, capsule, injection, granule, gel ointment, liniment or patch.
7. Use according to claim 5, characterized in that: the cosmetic or daily chemical product is in the form of gel ointment, liniment, patch, cream, detergent, emulsion, gel or toner.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112316110A (en) * | 2020-11-12 | 2021-02-05 | 温州大学 | Pharmaceutical preparation for promoting skin wound repair and preparation method thereof |
| CN114853852A (en) * | 2022-05-25 | 2022-08-05 | 四川大学 | Polypeptide and application thereof in promoting bone repair |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009209064A (en) * | 2008-03-03 | 2009-09-17 | Nara Institute Of Science & Technology | Peptide having cell proliferation-promoting activity |
| CN106699849A (en) * | 2015-11-11 | 2017-05-24 | 暨南大学 | Novel polypeptide and Periplaneta americana total polypeptide extract capable of promoting tissue repair, and application thereof |
| CN106977586A (en) * | 2016-01-15 | 2017-07-25 | 暨南大学 | American cockroach novel polypeptide and application that a kind of promotion organization is repaired |
-
2020
- 2020-03-04 CN CN202010142284.3A patent/CN111269292B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009209064A (en) * | 2008-03-03 | 2009-09-17 | Nara Institute Of Science & Technology | Peptide having cell proliferation-promoting activity |
| CN106699849A (en) * | 2015-11-11 | 2017-05-24 | 暨南大学 | Novel polypeptide and Periplaneta americana total polypeptide extract capable of promoting tissue repair, and application thereof |
| CN106977586A (en) * | 2016-01-15 | 2017-07-25 | 暨南大学 | American cockroach novel polypeptide and application that a kind of promotion organization is repaired |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112316110A (en) * | 2020-11-12 | 2021-02-05 | 温州大学 | Pharmaceutical preparation for promoting skin wound repair and preparation method thereof |
| CN112316110B (en) * | 2020-11-12 | 2023-06-23 | 温州大学 | Pharmaceutical preparation for promoting skin wound repair and preparation method thereof |
| CN114853852A (en) * | 2022-05-25 | 2022-08-05 | 四川大学 | Polypeptide and application thereof in promoting bone repair |
| CN114853852B (en) * | 2022-05-25 | 2023-04-25 | 四川大学 | Polypeptides and their use in promoting bone repair |
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