CN109912685A - Eye protection wins peptide and combinations thereof and the purposes using the victory peptide - Google Patents
Eye protection wins peptide and combinations thereof and the purposes using the victory peptide Download PDFInfo
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- CN109912685A CN109912685A CN201711326809.3A CN201711326809A CN109912685A CN 109912685 A CN109912685 A CN 109912685A CN 201711326809 A CN201711326809 A CN 201711326809A CN 109912685 A CN109912685 A CN 109912685A
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- peptide
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Abstract
The present invention relates to eye protection victory peptide and combinations thereof and use the purposes of the victory peptide.Specifically, the present invention provides a kind of synthesis victory peptide, is made of the amino acid sequence of sequence ArgAsnProLeuGluGluThr (SEQ ID NO:1).There is provided one simultaneously includes the victory peptide to eye protection or the helpful medical composition of eye health and application thereof.
Description
Technical field
The present invention relates to widely effectively win peptide, its composition and the purposes using the victory peptide to protection eye health.
Background technique
There is no eyes, we can not operate computer, see TV, book or newspaper reading, drive vehicle and play chess.Recently, mostly
Number people takes a lot of time through non-moving or mobile device, such as television set, computer, desktop computer, laptop, puts down
Plate computer, mobile phone etc. watch film, cause the damage of vision or a series of symptoms such as eyes it is dry and astringent, it is uncomfortable, stimulation, it is scorching hot,
Rubescent, excessive tearing eye-blurred, fatigue, aches, anxiety, photophobia, myopia, astigmatism.
There is any damage to eyes in order to prevent, has and develop many methods in different ways.For example, by sleeping,
Doze, movement, music, long sight, massages eyes, wears sunglasses at eye closing, takes vitamin, eye drops etc., can easily alleviate
Eye fatigue.The method for separately thering are other to reduce eye irritation.
Needed for meeting eye health, drug such as vitamin or herbal medicine or combinations thereof object has been suggested.But to being at present
Only, it is still uncertain for being beneficial to the drug of eye health.
We remain desirable to that nontoxic, nonantigenic and a not expensive eye health is found or developed for eye health
Agent.
Summary of the invention
In the present invention surprisingly, there is ArgAsnProLeuGluGluThr (SEQ ID NO:1) amino acid sequence
The victory peptide of column has the effectiveness for promoting corneal epithelial cell migration, helpful to eye protection or eye health.
Therefore, the amino acid sequence that the present invention provides that synthesis victory peptide includes SEQ ID NO:1 on the one hand is formed,
It is named as No. 12 victory peptides.This victory peptide provides the effectiveness for promoting corneal epithelial cell migration.
On the other hand, the medical composition helpful the present invention provides a pair of eyes protection or eye health, it includes
A effective amount of pharmaceutically acceptable carrier of SEQ ID NO:1 and one.
One embodiment of the present of invention, its composition are modulated into a kind of system or local application form.Another
In specific embodiment, the composition is modulated into the form of local application, such as the drops of eye.
On the other hand, the present invention provides the method for the eye protection of a prevention ocular damage, it includes in need
Individual application have SEQ ID NO:1 amino acid sequence victory peptide, usage amount be effectively promote corneal epithelial cell migration
Effectiveness amount.
One embodiment according to the method for the present invention, victory peptide are locally applied to individual.
It should be appreciated that both previous general description and detailed description below are all merely illustrative and explanatory and be not intended to limit this
Invention.
Detailed description of the invention
The previous general introduction of the present invention and it is described below cooperate annexed drawings read when be more preferably appreciated that.To say
The bright present invention shows the currently preferred embodiment in the accompanying drawings.
In the accompanying drawings:
Figure 1A and Figure 1B shows the migration of No. 12 victory peptide promoted corneal epithelial cell HCEC at 24 hours；Wherein Figure 1A is mentioned
For detecting the representative figure in No. 12 victory peptide cultures 24 hours (scale bar, 1mm) of various concentration using open-work migration.Figure 1B is provided
Statistic analysis result (* P < 0.05) of the open-work migration detection at 24 hours.
Fig. 2A and Fig. 2 B shows the migration of No. 12 victory peptide promoted corneal epithelial cell HCEC at 48 hours；Wherein Fig. 2A is mentioned
For detecting the representative figure in No. 12 victory peptide cultures 48 hours (scale bar, 1mm) of various concentration using open-work migration.Fig. 2 B is provided
Statistic analysis result (* * P < 0.01) of the open-work migration detection at 48 hours.
Fig. 3 shows that survival ability of No. 12 victory peptides for HCEC cell at 24 hours has no significant effect, wherein utilizing MTT
Test is by HCEC cell at No. 12 victory peptides culture 24 hours of various concentration.
Fig. 4 shows that survival ability of No. 12 victory peptides for HCEC cell at 48 hours has no significant effect, wherein utilizing MTT
Test is by HCEC cell at No. 12 victory peptides culture 48 hours of various concentration.
Specific embodiment
Unless otherwise defined, all technologies used herein and scientific term have with it is common in fields of the present invention
Technical staff understands identical meaning.
When term "a" or "an" is used together in claims and/or specification with term "comprising", meaning
Think to be "one", but it is also consistent with the meaning of " one or more ", "at least one" and " one or more in one ".
Term " victory peptide " is herein defined as being used with its conventional sense, and also as a kind of polymer, monomer are ammonia
Base acid, and interlinked together by amido bond, separately selectively, refer to polypeptide (polypeptide).When the amino
When acid is a-amino acid, L- optical isomeric compound or D- optical isomeric compound can be used.In addition, also may include non-natural amino acid,
For example, Beta-alanine, phenylglycine and homoarginine.Use the standardized abbreviations of amino acid.
Term " individual " used herein refers to vertebrate, preferably mammal, it is further preferred that ground is people.Under
Wen Zhong, the people as individual are especially referred to as " human individual ".
Term " carrier " used herein refer to be generally used for modulation can promote stability, aseptic, delivering property
The material of drug or cosmetic composition.When winning delivery of peptides system modulation is a kind of solution or suspension, which is
In an acceptable carriers, preferably aqueous carrier.A variety of aqueous carriers can be used, for example, water, buffer, 0.8% salt
Water, 0.3% glycine, hyaluronic acid etc..The composition contains as required and is close to can physiologically connecing for physiological condition
The auxiliary substance received, such as pH adjustment and buffer, solution tension regulator, wetting agent and the like, for example, sodium acetate,
Sodium lactate, sodium chloride, potassium chloride, calcium chloride, sorbitanmonolaureate, triethanolamine oleate etc..
Term " system " " systematically " in this article refers to the way that drug or other materials are applied to the circulatory system
Diameter, so that whole bodies of individual are influenced by applying.The application can by enteral administration (absorbed the drug by gastrointestinal tract or its
His substance) or parenteral administration as injected, infusion or be implanted into be administered.
Term " local " " locally " is herein defined as being used with its conventional sense, and referring to can be in body
Position in or on any position, including but not limited to epidermis, any other corium or any other bodily tissue.Part
Ground, which is applied or bestowed, means to make the victory peptide directly to contact with tissue, such as skin or film containing melanin production cell.
Term " effective quantity " in this article refers to the victory peptide of sufficient amount, provides required treatment or beauty according to the present invention
Effect or the certain types of reaction of induction.A effective amount of demand is variant between individuals, is state according to disease, physics shape
Condition, age, gender, the type of individual and weight etc..However effective quantity appropriate can generally make by standard method or with any
With or mode well known by persons skilled in the art synthesize.For example, victory peptide according to the present invention can systematically, percutaneous or part is applied
With.
As used herein term " pharmaceutically acceptable carrier " refers to any standard pharmaceutical carriers.The carrier packet
Contain, but be not limited to: normal saline solution, buffered saline, glucose, water, glycerol, ethyl alcohol, propylene glycol, polyoxyethylene castor
Sesame oil, nanoparticle, liposome, polymer and combinations thereof.Other than standard vector, medical composition of the invention by one or
A variety of excipient being usually used in typical standard preparation, for example, surfactant, solubilizer, stabilizer, emulsifier, thickener and
Preservative.This excipient is well known to those skilled in the art.
As shown in the examples, win peptide composed by the amino acid sequence with SEQ ID NO:1, it can be by standard method
By it is any generally use or it is well known by persons skilled in the art in a manner of synthesize.It is proved promote horn cell expression and
The migration of fibroblast is effective.Therefore, the present invention also provides a kind of method for promoting wound healing, and it includes in need
Individual application there is the victory peptide of SEQ ID NO:1 (while being named as No. 12 victory peptides) amino acid sequence, usage amount is effectively to increase
Into the amount of the migration of corneal epithelial cell.
On the other hand, the present invention, which is provided, manufactures the medicine that a pair of eyes is protected or eye health is beneficial using invention victory peptide
Upper or health protection composition.
In addition, the present invention provides a kind of pair of eye protection or the helpful composition of eye health or medical composition,
Include a effective amount of pharmaceutically acceptable carrier of SEQ ID NO:1 and one.
Medical composition of the invention can be by one or more pharmaceutically acceptable carriers to be suitable for selected administration
Mode is applied, and including system or local application, passes through enteral or parenteral administration such as injection, infusion or implantation, oral, warp
Skin or local administration.Section Example of the invention, its composition are prepared as pharmaceutically or cosmetically acceptable carrier, preparation
Include liquor, ointment, gel, slurries, creams, lotion, powder and latex and any form applied.In section Example
In, said preparation is applied by way of drops at a glance.
Be further illustrated by the examples that follow the present invention, the embodiment be with provide illustrative purpose provide rather than
Limitation.
Embodiment
The preparation of embodiment 1:12 victory peptide
The victory peptide of SEQ ID NO:1 (sequence: ArgAsnProLeuGluGluThr) be by Sheng Gong Co., Ltd (MDBio,
Inc.) (Taibei, TaiWan, China) is synthesized, and by high-effect liquid chromatography (HPLC) and mass spectrography confirmation win peptide purity and
Composition.After 10mg freeze-drying victory peptide powder is dissolved in the 250 bis- deionized waters of μ l (ddH2O), victory peptide raw material is stored in -20
℃。
Embodiment 2: Efficacy experiments
Materials and methods
Cell culture
Human corneal epithelial cell HCEC is at 37 DEG C and contains 5%CO2Under, be incubated at containing keratinocyte without blood
Clear culture medium, the Niu Chuiti extract of human recombinant EGF, 0.05mg/ml of 5ng/ml, 0.005mg/ml insulin and 500ng/
The cortisol of ml.
Open-work migration detection
By the cell (3.5 × 10 in the serum-free medium of 0.25ml keratinocyte4) with the film in 8 μm of apertures
(Corning, USA) is planted in upper chamber.The serum-free KSF for containing or not contain No. 12 victory peptides is loaded onto the hole of 24 well culture plates
In lower room in.After culture 24 hours or 48 hours, the cells are fixed and is tied with 0.5% (w/v) containing 20% (v/v) methanol
Crystalviolet (Sigma) dyeing.The migrating cell number from 5 random fields is counted under phase contrast microscope.Its result is with student t inspection
Analysis, and it is plotted as average value ± standard deviation.The result is obtained from two independent experiments.
MTT detection
By the cell (2 × 10 in complete KSF4) kind is in containing or not contain in No. 12 victory 12 orifice plates of peptide.Into each hole
70 microlitres of 5mg/ml 3- (4,5- dimethyl -2- thiazolyl) -2,5- diphenyl -2H- tetrazolium bromide solution (MTT is added；
Sigma it), and at 37 DEG C cultivates 3 hours.After that, 10%SDS of the 700 μ l in 0.01N HCl is added to dissolve MTT
FirstCrystallization.The gained optical density under the dual wavelength of 550 and 630nm is measured with light splitting luminance meter.
As a result
No. 12 victory peptides of concentration of the human corneal epithelial cell HCEC through 5,10,25,50 μ g/ml are handled 24 and 48 hours,
Then it is tested and analyzed using open-work migration.If Figure 1A, Figure 1B, Fig. 2A, Fig. 2 B are shown, No. 12 victory peptides enhance corneal epithelial cell
The migration of HCEC.However, Fig. 3 and Fig. 4 show No. 12 victory peptides 5,10,25,50 μ g/ml concentration, 24 and 48 hours, for
The survival ability of HCEC cell has no significant effect.
In view of above-mentioned, infer that the victory peptide of SEQ ID NO:1 by the migration of enhancing corneal epithelial cell, rather than passes through angle
The viability of film epithelial cell.
It will be understood by those skilled in the art that above-mentioned tool can be modified in the case where not departing from broad inventive concept of the invention
Body embodiment.Accordingly, it will be understood that the invention is not limited to revealed certain specific embodiments, and it is intended to cover
Modification in the spirit and scope of the present invention as defined in the appended claims.
Sequence table
<110>three all raw skills research and develop limited liability company
<120>eye protection victory peptide and combinations thereof and the purposes using the victory peptide
<130> ASB0004WO
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 7
<212> PRT
<213>artificial sequence
<400> 1
Arg Asn Pro Leu Glu Glu Thr
1 5
Claims (10)
- It is by the amino acid sequence institute group of ArgAsnProLeuGluGluThr (SEQ ID NO:1) 1. a kind of synthesis wins peptide At.
- 2. synthesis according to claim 1 wins peptide, has effects that a migration that can effectively promote corneal epithelial cell.
- 3. a kind of pair of eye protection or the helpful composition of eye health or medical composition, it includes a effective amount of such as right It is required that synthesis described in 1 wins peptide, the synthesis victory peptide is that the amount for the migration for effectively promoting strong fibroblast and one pharmaceutically may be used The carrier of receiving.
- 4. according to claim 3 to eye protection or the helpful composition of eye health or medical composition, quilt It is modulated into a kind of system or local application preparation.
- 5. according to claim 3 to eye protection or the helpful composition of eye health or medical composition, quilt It is modulated into a kind of preparation of local application.
- 6. it is according to claim 4 to eye protection or the helpful composition of eye health or medical composition, wherein The preparation of the local application is drop form at a glance.
- 7. a kind of purposes for preventing ocular damage for eye protection, it includes can effectively increase to an individual application in need A effective amount of synthesis as described in claim 1 into the migration of corneal epithelial cell wins peptide.
- 8. purposes according to claim 7, wherein as described in claim 1 synthesis victory peptide can system or part it is right Individual application.
- 9. purposes according to claim 8, wherein synthesis victory peptide can locally apply individual as described in claim 1 With.
- 10. purposes according to claim 8, wherein synthesis victory peptide is modulated into a kind of eye as described in claim 1 Drop form.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711326809.3A CN109912685B (en) | 2017-12-13 | 2017-12-13 | Eye protection peptide, composition thereof and application of eye protection peptide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711326809.3A CN109912685B (en) | 2017-12-13 | 2017-12-13 | Eye protection peptide, composition thereof and application of eye protection peptide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109912685A true CN109912685A (en) | 2019-06-21 |
| CN109912685B CN109912685B (en) | 2022-05-06 |
Family
ID=66958543
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711326809.3A Active CN109912685B (en) | 2017-12-13 | 2017-12-13 | Eye protection peptide, composition thereof and application of eye protection peptide |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0914827A1 (en) * | 1996-06-26 | 1999-05-12 | Santen Pharmaceutical Co., Ltd. | Ophthalmic drug compositions |
| US20110059504A1 (en) * | 2008-01-03 | 2011-03-10 | Getmanova Elena V | Engineered transglutaminase barrel proteins |
| WO2016179007A1 (en) * | 2015-05-01 | 2016-11-10 | Allysta Pharmaceuticals, Inc. | Adiponectin peptidomimetics for treating ocular disorders |
| WO2019061491A1 (en) * | 2017-09-30 | 2019-04-04 | Pro Sunfun Biotech Research And Development Co., Ltd. | Eye-care peptide, its composition and method of using the same |
| CN112823789A (en) * | 2019-11-19 | 2021-05-21 | 强生外科视力公司 | Compositions and methods for treating the eye |
-
2017
- 2017-12-13 CN CN201711326809.3A patent/CN109912685B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0914827A1 (en) * | 1996-06-26 | 1999-05-12 | Santen Pharmaceutical Co., Ltd. | Ophthalmic drug compositions |
| US20110059504A1 (en) * | 2008-01-03 | 2011-03-10 | Getmanova Elena V | Engineered transglutaminase barrel proteins |
| WO2016179007A1 (en) * | 2015-05-01 | 2016-11-10 | Allysta Pharmaceuticals, Inc. | Adiponectin peptidomimetics for treating ocular disorders |
| WO2019061491A1 (en) * | 2017-09-30 | 2019-04-04 | Pro Sunfun Biotech Research And Development Co., Ltd. | Eye-care peptide, its composition and method of using the same |
| CN112823789A (en) * | 2019-11-19 | 2021-05-21 | 强生外科视力公司 | Compositions and methods for treating the eye |
Non-Patent Citations (2)
| Title |
|---|
| QING YUAN等: "Protective efficacy of a peptide derived from a potential adhesin of Pseudomonas aeruginosa against corneal infection", 《EXPERIMENTAL EYE RESEARCH》 * |
| 王东波等: "基因重组PACAP27衍生多肽RP2制备及促角膜上皮细胞增殖研究", 《中国生物工程杂志》 * |
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| Publication number | Publication date |
|---|---|
| CN109912685B (en) | 2022-05-06 |
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