CN102458357A - Collagen production enhancer, and external composition for skin and cosmetic each comprising same - Google Patents
Collagen production enhancer, and external composition for skin and cosmetic each comprising same Download PDFInfo
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- CN102458357A CN102458357A CN2010800282277A CN201080028227A CN102458357A CN 102458357 A CN102458357 A CN 102458357A CN 2010800282277 A CN2010800282277 A CN 2010800282277A CN 201080028227 A CN201080028227 A CN 201080028227A CN 102458357 A CN102458357 A CN 102458357A
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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
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
- A61K8/981—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of mammals or bird
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/16—Emollients or protectives, e.g. against radiation
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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
- A61Q19/004—Aftersun preparations
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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
- A61Q19/08—Anti-ageing preparations
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Abstract
Disclosed is a collagen production enhancer, which comprises an enzymatically decomposed product of swallow's nest as an active ingredient. Specifically disclosed are: an excellent type-I collagen production enhancer which can be produced with high efficiency and is highly safe; and an external preparation and a cosmetic, each of which comprises the type-I collagen production enhancer.
Description
The reference of related application
The application is willing to 2009-148754 number based on the japanese patent application laid that proposes earlier, and (applying date: on June 23rd, 2009), and advocate the interests of its priority, its disclosure integral body is incorporated herein through reference.
Technical field
The present invention relates to the zymolyte of Nidus collocaliae collagen production promoter as effective ingredient.More specifically, the present invention relates to promote the collagen production promoter of the type i collagen generation in the dermis of skin fibroblast.In addition, the present invention relates to comprise composition for external application and cosmetics as the zymolyte of the Nidus collocaliae of its effective ingredient.
Background technology
Skin problems such as lax, the elastic reduction of the coarse and/or wrinkle of skin, skin particularly are the thing of important care for the women.Think what these skin problems were normally caused by psychological factors such as environmental factors, the factor of following age growth such as aging and pressure etc.That particularly, we can say that dryingization by the living environment of following ultraviolet increase in recent years, a large amount of uses of air-conditioning etc. etc. causes brings dysgenic environmental factors to increase to skin.
Skin histology comprises epidermis, corium, subcutaneous tissue.In corium, comprise collagen, hyaluronic acid in large quantities, water-retaining property, the elasticity of they and skin are closely related.The dermal fibroblast that exists in the known corium produces collagen, hyaluronic acid.
It is generally acknowledged in the protein that collagen exists to account for 30% that it 70% is present in the corium in people's organism.As the collagen that is present in corium, comprise type i collagen, III Collagen Type VI, but mainly be type i collagen.
In the past, for the elasticity that improves skin and/or gloss, prevention improve wrinkle, adopt used the cosmetics that cooperated collagen with the mode of external more, directly replenished the method for collagen to skin.Yet, replenishing under the situation of collagen with the mode of external, owing to be difficult to make the collagen quantity in the dermis of skin to increase, therefore for the elasticity that improves skin and/or gloss, prevention improved wrinkle, not talkative must be sufficient.
Comprise in the ultraviolet light if known skin is exposed to, the most collagen that then occupies the extracellular matrix of corium reduces significantly, and the at random of its fibre bundle takes place.Therefore; Making the collagen quantity of corium increase improvement for wrinkle, prevent etc. is useful (document 1 (" the plain material development of function make-up article め experiment method (experimental method that is used for the development functionality cosmetic raw materials) " (taro Pi Xuaner chief editor); Distribution in 2007; シ one エ system シ one publishes the 45th～47 page)).In addition, in the document, demonstrate, through the irradiation of ultraviolet B radiation (UVB), the collagen of dermal fibroblast is synthetic to be reduced significantly.In the document, as the example of type i collagen synthesis accelerant, illustration vitamin C derivatives, milk surum become to grade.
On the other hand, at document 2 (Akiko Kojima-Yuasa, et al.; Fragrance Journal, April, 2009; (Vol.37 No.4), demonstrates in pp.103-) No.346; It is useful suppressing that reduction that collagen produces ability improves for the raising of QOL, and the reduction of collagen quantity becomes wrinkle, lax etc. reason in the skin.
As the generation promoter of type i collagen, up to now, reported the generation promoter (for example, TOHKEMY 2008-105984 communique, WO2004/085429) of the type i collagen that is conceived to derive from the composition of specific plant and utilized this composition in large quantities.
Nidus collocaliae is that cave swallow (ア Na Star バ メ) makes the saliva of oneself become thread and makes nest, and as its composition, the known glycoprotein that comprises in a large number contains lipid hardly.In the past, in the edible raw material of Chinese dishes, the raw material of Chinese medicine etc., used Nidus collocaliae.In recent years, also developed Nidus collocaliae as the health food of raw material etc., but only known the suitable example of only a few the such external purposes of external agent, cosmetics.
In TOHKEMY 2003-95961 communique, the for oral use U.S. flesh promoter of using the zymolyte of Nidus collocaliae is disclosed.Yet, in the document, only only shown oral use, both do not put down in writing hint about the probability of utilizing in the external purposes yet.In addition, the effect here is to improve the moist feeling of skin, obtain the thin skin of texture, promotes ability about the generation with the type i collagen of the improvement of the elasticity of skin and/or glossy raising, wrinkle, has no record.
In WO99/022709, the aqueous solvent extract, the particularly hot water extract's that comprise Nidus collocaliae cosmetics are disclosed.These cosmetics can be used for the skin nursing purposes, are the cosmetics that in the external purposes, use.In addition, in the document, the aqueous extract that also discloses Nidus collocaliae shows the synthetic facilitation effect of collagen in the Skin Cell.Yet employed here extract is that Nidus collocaliae is carried out hot water extraction, has removed the extract behind the solid constituent, and the Nidus collocaliae zymolyte that directly uses with Nidus collocaliae integral body being carried out enzyme handle is obviously different.In addition, about effect, in the inventor's consideration limit, thinking has room for improvement.
Summary of the invention
The inventor finds that specifically the zymolyte of Nidus collocaliae has the generation facilitation effect of the excellence of the type i collagen in the dermis of skin fibroblast.This effect in addition, is further being carried out in the zymolyte of enzymolysis than the more remarkable effect that in the hot water extract of Nidus collocaliae, shows, and this effect is more remarkable.The present invention is based on these and is familiar with and accomplishes.
Therefore, the purpose of this invention is to provide and can produce efficiently and the generation promoter of safe, excellent type i collagen, and the external agent and the cosmetics that comprise the generation promoter of this type i collagen.
Collagen production promoter of the present invention with the zymolyte of Nidus collocaliae as effective ingredient.
According to an optimal way of the present invention, above-mentioned zymolyte is the protease hydrolyzed thing.
According to a more preferably mode of the present invention, the molecular weight of above-mentioned zymolyte is 70～200,000.
According to a further optimal way of the present invention, the mean molecule quantity of above-mentioned zymolyte is 300～15,000.
According to optimal way of the present invention, collagen production promoter of the present invention is used for promoting that the type i collagen of dermis of skin fibroblast produces.
According to alternate manner of the present invention, the composition for external application that comprises collagen production promoter of the present invention is provided.Said composition is preferred for skin nursing, more preferably is used to improve the elasticity and the gloss of skin and prevents or improve wrinkle.
According to other more preferably mode of the present invention, composition for external application of the present invention is cosmetics.
According to alternate manner of the present invention, the application of zymolyte of the Nidus collocaliae of the collagen production promoter that is used to make the mammalian skin that comprises the people is provided.According to another mode of the present invention, the application of the zymolyte of the Nidus collocaliae that is used to make the composition for external application that skin nursing uses is provided.
According to alternate manner of the present invention, the generation promotion method of the collagen in the skin is provided, it comprises the mammalian skin that the zymolyte as effective ingredient of the present invention of effective dose is applied to comprise the people.According to another mode of the present invention, the skin nursing method is provided, it comprises the mammalian skin that the effective ingredient of the present invention of effective dose is applied to comprise the people.Likewise, also provide the elasticity and the gloss that make skin to improve, prevent or improve the method for wrinkle, it comprises the mammalian skin that the effective ingredient of the present invention of effective dose is applied to comprise the people.
According to the present invention, through with the zymolyte of Nidus collocaliae as effective ingredient, and compared in the past, can promote the generation of the collagen of application on human skin dermal fibroblast significantly.In addition, promote ability, zymolyte is used for external preparation for skin purposes or cosmetic use, thereby can improve the elasticity of skin, the gloss that improves skin and prevention or reduce wrinkle through generation based on the collagen of this dermal fibroblast.In addition, promote collagen to produce in the synthetic skin that has reduced such state of collagen in can the skin after Exposure to Sunlight, thus improve skin elasticity and/or gloss, improve wrinkle.Like this, according to the present invention, can seek the raising of the QOL of consumer.In addition, therefore safe because employed zymolyte uses the well-known Nidus collocaliae as eating raw material, consumer also is easy to accept.In addition, handle and obtain zymolyte, therefore can make efficiently, and can also suppress manufacturing cost lower owing to can Nidus collocaliae directly be carried out enzyme.
Description of drawings
Fig. 1 is the figure that shows the mensuration result of the sialic acid amount in the Nidus collocaliae sample.
Fig. 2 is the figure that shows the mensuration result of the albumen quality in the Nidus collocaliae sample.
Fig. 3 is the figure that shows the mensuration result utilize the Nidus collocaliae zymolyte 1 that GPC-HPLC obtains.
Fig. 4 is the figure that shows the mensuration result utilize the Nidus collocaliae zymolyte 2 that GPC-HPLC obtains.
Fig. 5 is the figure that the generation of demonstration collagen promotes the mensuration result of ability.
The specific embodiment
Effective ingredient
About collagen production promoter of the present invention, as stated, with the zymolyte of Nidus collocaliae as effective ingredient.
As the Nidus collocaliae of the raw material of zymolyte is that the cave swallow makes the saliva of oneself become thread and makes nest, as its composition, comprises glycoprotein in a large number, comprises lipid hardly.Here, the cave swallow be meant belong to Apodidae (
Apodidae) esculent swift belong to (
Collocalia) the swallow class, for example can enumerate, the little grey waist esculent swift of Apodidae (
Apodidae Collocalia Francica), Apodidae tongue nest esculent swift (
Apodidae Collocalia Salangana), Apodidae Java esculent swift (
Apodidae Collocalia Fuciphaga), Apodidae ash waist esculent swift (
Apodidae Collocalia Inexpectata), the brown waist esculent swift of Apodidae (
Apodidae Collocalia Vestita), swift actions and spoken parts in classical Chinese drama abdomen esculent swift (
Apodidae Collocalia Esculenta), swift University of Science and Technology esculent swift (
Apodidae Collocalia Maximus)Deng.
In the present invention, Nidus collocaliae can use commercially available article.In usually commercially available Nidus collocaliae; Have from only removing dirts such as QUMAO, excrement and washing Nidus collocaliae to the particle of collecting Nidus collocaliae and a plurality of kinds such as Nidus collocaliae of bleaching repeatedly and washing and forming, but the used Nidus collocaliae of the present invention preferably uses the Nidus collocaliae that in pre-treatment, does not carry out over-drastic washing, bleaching etc.
The zymolyte of Nidus collocaliae be meant use that Nidus collocaliae is decomposed in enzyme reaction and material, can decompose through the compound enzyme etc. that uses protease, comprises protease and obtain Nidus collocaliae or its handled thing.Enzyme as using in the resolution process here is preferably protease, more preferably pancreatin.
Usually can handle through at first implementing pasteurization, in water, carry out enzyme then and handle the zymolyte that obtains Nidus collocaliae.Can suitably set the condition of sterilization processing, can also come suitably to set the condition that enzyme is handled according to the kind of employed enzyme, the state of Nidus collocaliae with reference to habitual pasteurization condition.
If enumerate example more specifically, then can be like the zymolyte of modulated Nidus collocaliae.
For example, add 10～50 times water of its quality in the Nidus collocaliae after pulverizing and make its swelling, handled 5 seconds～30 minutes at 60～130 ℃ of following pasteurizations then.Then, directly add an amount of enzyme, under the best pH of enzyme, optimum temperature, carried out enzyme reaction 0.5～48 hour.After enzyme reaction finishes, make enzyme deactivation through heat treated etc., filtering reacting liquid and remove insoluble matter, thus can obtain the zymolyte of Nidus collocaliae.In addition, this zymolyte drying and powdered as required.In addition, in each above-mentioned operation, can carry out operations such as suitable pH adjustment, decolouring, deodorize.
As other example; Under situation with the handled thing enzymolysis of Nidus collocaliae, to pulverize for below the particle diameter 2mm, be preferably 10～100 times the water that adds its quality in the Nidus collocaliae below the 150 μ m, under 1～60 ℃, leave standstill or stir and made its swelling in 0.5～48 hour; Handled 5 seconds～30 minutes at 60～130 ℃ of following pasteurizations then; Filter as required, the solution of gained and the above-mentioned enzyme that likewise carries out are handled, thereby can obtain the zymolyte of Nidus collocaliae.
In the modulation method of zymolyte, the time of enzyme reaction is long more, promotes proteinic decomposition more, obtains to comprise more low molecular proteinic zymolyte.In the present invention, preferably making the enzyme reaction time is about 0.5～48 hour, is preferably about 3～36 hours, more preferably about about 24 hours, and the zymolyte that has further carried out enzyme reaction.
The molecular weight of the zymolyte of the Nidus collocaliae that obtains like this is preferably 70～200, and 000, more preferably 70～180,000.Further preferred molecular weight is 70～150,000, further more preferably 70～120,000.
In addition, the mean molecule quantity (weight average molecular weight) of the zymolyte of expectation Nidus collocaliae is 300～100,000, is preferably 300～70,000.Preferred mean molecule quantity is 300～50,000, further is preferably 300～30,000, further more preferably 300～20,000, be preferably 300～15,000 especially.
Composition for external application
Collagen production promoter of the present invention can directly use individually, is contained in the various composition for external application such as cosmetics, medicine, quasi drug but also can be used as additive-package, can obtain to have the compositions that collagen produces facilitation effect.The compositions of gained can be used for skin nursing effectively, preferably is used to improve the elasticity and the gloss of skin effectively and prevents or improve wrinkle.
The generation that has the type i collagen in the application on human skin dermal fibroblast as the zymolyte of the Nidus collocaliae of effective ingredient of the present invention promotes active (result of the test of the embodiment that afterwards states).
Therefore, in the present invention, the generation of collagen promotes the active effect of generation ability that promotes or improve fibroblastic type i collagen of application on human skin corium that is meant, means the collagen quantity that can increase in the application on human skin corium thus.Can obtain by the collagen quantity that increases corium wrinkle improvement, prevent effect and can improve the elasticity of skin, the gloss that improves skin is that those skilled in the art are well-known, this point is (for example, above-mentioned document (" the plain material development of function make-up article め experiment method (experimental method that is used for the development functionality cosmetic raw materials) " (taro Pi Xuaner chief editor); Distribution in 2007, シ one エ system シ one publishes the 45th～47 page (non-patent literature 1)), document (AkikoKojima-Yuasa; Et al., Fragrance Journal, April; 2009; Demonstrate clearly among No.346 (Vol.37, No.4), pp.103-(non-patent literature 2)) etc.Therefore; Since confirmed effective ingredient of the present invention have collagen in the corium produce to promote effective ingredient of the present invention active, therefore clear and definite skin nursing, preferably the elasticity of skin and glossy raising and wrinkle prevent or improve in be effective.
In addition, in this manual, " prevention or improve " of skin such as the elasticity of skin and/or gloss, wrinkle or skin condition used with the meaning of the adjusting that comprises this state, the delay of carrying out, mitigation, prevention outbreak, prevention of recurrence, inhibition etc.
Therefore, as stated,, the composition for external application that comprises collagen production promoter of the present invention is provided according to alternate manner of the present invention.Preferably provide composition for external application as medicine, quasi drug or cosmetics, more preferably provide composition for external application as cosmetics.
In composition for external application of the present invention (particularly cosmetics); As the restriction especially of content (dry conversion) of the Nidus collocaliae zymolyte of effective ingredient, different and different according to the form of compositions, but generally speaking; As solid constituent; Be in the scope of 0.001～20 weight %, be preferably in the scope of 0.005～5 weight %, can suitably change according to the form of compositions.
Perhaps, the content of the Nidus collocaliae zymolyte in the composition for external application of the present invention (dry conversion) is preferably with more than the every day 0.01mg, more preferably 0.01mg～2mg, the mode that can be applied to the skin of per 1 square of cm comprise.
As long as composition for external application of the present invention comprises effective ingredient, and do not damage the generation facilitation effect of collagen, promoter then of the present invention can further comprise other any composition.As so any composition; For example can enumerate; The various compositions of habitually practising in skin preparations for extenal use, the cosmetics; For example, wetting agent, lower alcohol, polyhydric alcohol, saccharide, surfactant, buffer agent, emulsifying agent, stabilizing agent, thickening agent, antioxidant, anticorrosion anti-microbial inoculum, chelating agen, pH regulator agent, spice, pigment, UV absorbent, ultraviolet scattering diluent, vitamins, amino acids, antiinflammatory, water, peptide, sugar alcohols, enzyme, plant extract class, antioxidant class, Talcum, clay, pollen, Margarita etc.
Here,, for example can enumerate acid mucopolysaccharideses such as Polyethylene Glycol, polypropylene glycol, glycerol, propylene glycol, Sorbitol, hyaluronic acid, chondroitin sulfate, aminoacid, collagen, elastin laminin etc. as wetting agent.
As lower alcohol, for example can enumerate ethanol, propanol, isopropyl alcohol etc.
As polyhydric alcohol, for example can enumerate glycerol, tetramethylolmethane, dipentaerythritol, ethylene glycol, propylene glycol, polypropylene glycol, 1,3 butylene glycol etc.
As saccharide, for example can enumerate saccharides such as glucose, maltose, lactose, D-glucuronic acid, sucrose, D-Sorbitol, anhydrosorbitol, cellulose, starch, oligosaccharide class, polysaccharide and their derivant etc.
About composition for external application of the present invention; Collagen production promoter through comprising the Nidus collocaliae zymolyte is adjusted according to the known method proper fit with these any compositions, can process habitual goods form in the various cosmetics such as lotion (lotion), emulsion, cream (cream), moisture retention liquid, face film agent, astringent, cleansing milk, refreshing body water, profit body frost.Composition for external application of the present invention can also be used as color make-up cosmetics such as foundation cream class, lip pomade, eye shadow, rouge; Cosmeceuticals such as Sun block goods, deodorant cosmetics; Shampoo, hair conditioner, Hair grooming cosmetics such as hair oil, are bathed bath with preparation and are applied some make up etc. at skin detergent.
According to alternate manner of the present invention, as stated, the application of zymolyte of the Nidus collocaliae of the collagen production promoter that is used to make the mammalian skin that comprises the people is provided.Here, the generation of the collagen in the skin promotes the preferred type i collagen generation that promotes in the dermis of skin fibroblast.
According to another mode of the present invention, as stated, the application of the zymolyte of the Nidus collocaliae that is used to make the composition for external application that skin nursing uses is provided.Here, composition for external application is preferred for improving elasticity and the gloss and the prevention of skin or improving wrinkle.In addition, composition for external application is preferably cosmetics.
According to alternate manner of the present invention, as stated, the generation promotion method of the collagen in the skin is provided, it comprises the mammalian skin that the zymolyte as effective ingredient of the present invention of effective dose is applied to comprise the people.In addition, " effective dose " is meant here, according to application, is sufficient amount for the generation that can bring into play collagen promotes activity in the desired regions in vivo.In addition, the generation of the collagen in the so-called here skin promotes to be preferably the type i collagen generation that promotes in the dermis of skin fibroblast.
In addition, according to another mode of the present invention, the skin nursing method is provided, it comprises the mammalian skin that the effective ingredient of the present invention of effective dose is applied to comprise the people.Likewise, also provide the elasticity and the gloss that make skin to improve, prevent or improve the method for wrinkle, it comprises the mammalian skin that the effective ingredient of the present invention of effective dose is applied to comprise the people.In addition, here " effective dose " is meant sufficient amount for obtaining required effect.
In addition, in this manual, used " pact " and " about " the expression of value mean, in order to reach the purpose of setting this value, comprise the variation of the admissible value of those skilled in the art.For example mean, value that can allow to stipulate or scope 20% with interior, be preferably 10% with interior, more preferably 5% with interior variation.
Embodiment
Based on following example the present invention is described particularly, but the present invention is not limited to these examples.
(1) modulation of Nidus collocaliae sample
(1-1) the Nidus collocaliae zymolyte 1
The Nidus collocaliae of commercially available nothing bleaching is pulverized with pulverizer; Obtain the Nidus collocaliae powder of 100 orders through (below the particle diameter 150 μ m); In this Nidus collocaliae powder, add the water of about 50 times of amounts (quality), 5 ℃ of following swellings 20 hours, then 121 ℃ of following pasteurizations processing 15 minutes.
The treatment fluid of cooling gained is adjusted pH then, and adding with respect to the Nidus collocaliae powder is the enzyme that contains protease (trade name " パ Application Network レ ア チ Application F ", Amono Pharmacevtical Co., Ltd.'s system) of the amount of 2 quality %, reacts 3 hours down at 45 ℃.
This enzyme reaction solution is adjusted to pH7.0, and heating made enzyme deactivation in 5 minutes under 90 ℃ then, filtered, and reclaimed filtrating.Concentrate and should filtrate, lyophilization then obtains " Nidus collocaliae zymolyte 1 ".
(1-2) the Nidus collocaliae zymolyte 2
Except the time that makes enzyme reaction is 24 hours, likewise carry out with the zymolyte 1 of above-mentioned (1-1), obtain " Nidus collocaliae zymolyte 2 ".
(1-3) Nidus collocaliae hot water extract (comparative example)
Nidus collocaliae (dry thing) 40g of commercially available nothing bleaching is pulverized with pulverizer, this ground product is added in the distilled water of 1000ml, under reflux, extract and handled 2 hours.After the reflux, leave standstill, filtrating is reclaimed in separation of supernatant and filtration then.With residue further with the above-mentioned distilled water that likewise is added into 1000ml in, under reflux, extract processing again, then the solid matter residue is removed through filtration, reclaim filtrating.The filtrating (extracting solution) that merges gained, concentrating under reduced pressure, the concentrate of lyophilization gained, " Nidus collocaliae hot water extract " (comparative example) of acquisition yellow-white.
(2) analysis of Nidus collocaliae sample
Sialic acid amount and albumen quality in each Nidus collocaliae sample (Nidus collocaliae zymolyte 1, Nidus collocaliae zymolyte 2 and Nidus collocaliae hot water extract) of following mensuration gained.
About sialic amount, after with each test portion acid hydrolysis, adopt HPLC, measure as free N-n acetylneuraminic acid n.
About proteinic amount, use protein determination test kit (Protein Assay Kit) mensuration based on the Bio Rad society of Brad ford method.
The result is as depicted in figs. 1 and 2.
Next, in order to measure Nidus collocaliae zymolyte 1 and proteinic molecular weight and distribution thereof that Nidus collocaliae zymolyte 2 is comprised, adopt the condition of following that kind to utilize GPC to analyze.
[condition determination of HPLC]
Post: (φ 7.6 * 300mm) for Shodex Asahipak GS320HQ
Column temperature: 35 ℃
Mobile phase: 50mM CH
3COONH
3PH6.7
Flow velocity: 0.4ml/ minute
Test portion sample size: 10 μ l
Detector: ultraviolet spectrometry detector (under UV220nm, measuring)
[molecular weight marker of use]
·BSA?MW?66,000
·Gly-gly?MW?132.1
·Gly-gly-gly?MW?189.17
L-aspartic acid MW133.1
DL-aspartic acid MW 150.1
N-n acetylneuraminic acid n MW 309.3
Measure result such as Fig. 3 and shown in Figure 4.
In addition, about the proteinic molecular weight that the Nidus collocaliae zymolyte is comprised, use western blotting to estimate.Particularly, at first,, confirm the proteinic amount in the Nidus collocaliae zymolyte according to above-mentioned algoscopy.Use SDS-PAGE, utilize 10～15% polyacrylamide to separate the protein (100 μ g) that derives from the Nidus collocaliae zymolyte.On protein transfer printing to the immunoblotting pvdf membrane (polyvinylidene fluoride is obtained by Bio-Lad Lab society) that is deployed on the gel.Pvdf membrane was sealed 1 hour in the PBS that comprises 3% defatted milk and 0.1% tween 20, visually judge.
In addition, when the evaluation of proteinic molecular weight, also with reference to known document (Guo, C.T., Suzuki Y., et al., data presented among the Antiviral Res.70, pp.140-146 (2006).
Can know that by these results the protein molecular weight that is comprised in the Nidus collocaliae zymolyte exists in the scope of about about 70～120,000 (in addition according to appreciation condition, higher limit is about 200,000 sometimes).
In addition, the result by gained can know that the proteinic mean molecule quantity in the Nidus collocaliae zymolyte 1 is about 55,000.Can know that in addition the proteinic mean molecule quantity in the Nidus collocaliae zymolyte 2 is about 33,000.
(3) generation of collagen promotes the mensuration of ability
Use contains the DMEM culture medium (being obtained by SIGMA) of 0.5% calf serum (FBS), on 96 orifice plates with 2 * 10
4The cell density inoculation normal human skin dermal fibroblast (KF-4109 is obtained by Network ラ ボ ウ society) of individual cell.After inoculating 24 hours,, cultivated again 24 hours with the DMEM culture medium exchange that contains 0.5%FBS of each the Nidus collocaliae sample that contains normal concentration.
In addition, as positive control (P.C.), use magnesium ascorbyl phosphate (VC-PMg).
After the cultivation, reclaim medium supernatant, measure the type i collagen amount in the supernatant through the ELISA method.In addition, in the ELISA method, the anti-anti-people's type i collagen antibody (rabbit) that uses, two anti-use Histofine PO (rabbit), and developer use 2,2 '-azine group two (3-ethyl benzo thiazole phenanthroline-6-sulfonic acid) (ABTS) utilizes ELIASA to measure the absorbance of 405nm.
Through with the albumen quality of the type i collagen amount in the culture medium, calculate the type i collagen amount in the culture medium as the type i collagen generation of per unit albumen quality divided by full cell.
The result is as shown in Figure 5.
Can know that by the result type i collagen that the zymolyte of Nidus collocaliae has increased the application on human skin dermal fibroblast significantly produces.
Claims (22)
1. collagen production promoter, its with the zymolyte of Nidus collocaliae as effective ingredient.
2. collagen production promoter according to claim 1, zymolyte are the protease hydrolyzed things.
3. collagen production promoter according to claim 1 and 2, the molecular weight of zymolyte are 70～200,000.
4. according to each described collagen production promoter of claim 1～3, the mean molecule quantity of zymolyte is 300～15,000.
5. according to each described collagen production promoter of claim 1～4, it is used for promoting that the type i collagen of dermis of skin fibroblast produces.
6. composition for external application, it comprises each described collagen production promoter of claim 1～5.
7. compositions according to claim 6, it is used for skin nursing.
8. according to claim 6 or 7 described compositionss, it is used to improve elasticity and the gloss and the prevention of skin or improve wrinkle.
9. according to each described compositions of claim 6～8, it is cosmetics.
10. the application of the zymolyte of a Nidus collocaliae is used to make the collagen production promoter of the mammalian skin that comprises the people.
11. application according to claim 10, the generation of the collagen in the skin promote it is to promote the type i collagen in the dermis of skin fibroblast to produce.
12. the application of the zymolyte of a Nidus collocaliae is used to make the composition for external application that skin nursing is used.
13. application according to claim 12, composition for external application are to be used to improve elasticity and the gloss and the prevention of skin or improve wrinkle.
14. according to each described application of claim 10～13, zymolyte is the protease hydrolyzed thing.
15. according to each described application of claim 10～14, the molecular weight of zymolyte is 70～200,000.
16. according to each described application of claim 10～15, the mean molecule quantity of zymolyte is 300～15,000.
17. according to each described application of claim 10～16, composition for external application is cosmetics.
18. the generation promotion method of the collagen in the skin, it comprises the mammalian skin that the zymolyte of the Nidus collocaliae of effective dose is applied to comprise the people.
19. method according to claim 18, the generation of the collagen in the skin promote it is to promote the type i collagen in the dermis of skin fibroblast to produce.
20. a skin nursing method, it comprises the mammalian skin that the zymolyte of the Nidus collocaliae of effective dose is applied to comprise the people.
21. elasticity and a gloss that improves skin, prevent or improve the method for wrinkle, it comprises the mammalian skin that the zymolyte of the Nidus collocaliae of effective dose is applied to comprise the people.
22. according to each described method of claim 18～21, zymolyte is the protease hydrolyzed thing.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610983140.4A CN107049907A (en) | 2009-06-23 | 2010-06-21 | The purposes of the pancreatin zymolyte of bird's nest |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP148754/2009 | 2009-06-23 | ||
| JP2009148754A JP5815202B2 (en) | 2009-06-23 | 2009-06-23 | Collagen production promoter, skin external composition containing the same, and cosmetics |
| PCT/JP2010/060459 WO2010150737A1 (en) | 2009-06-23 | 2010-06-21 | Collagen production enhancer, and external composition for skin and cosmetic each comprising same |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610983140.4A Division CN107049907A (en) | 2009-06-23 | 2010-06-21 | The purposes of the pancreatin zymolyte of bird's nest |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102458357A true CN102458357A (en) | 2012-05-16 |
Family
ID=43386506
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610983140.4A Pending CN107049907A (en) | 2009-06-23 | 2010-06-21 | The purposes of the pancreatin zymolyte of bird's nest |
| CN2010800282277A Pending CN102458357A (en) | 2009-06-23 | 2010-06-21 | Collagen production enhancer, and external composition for skin and cosmetic each comprising same |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610983140.4A Pending CN107049907A (en) | 2009-06-23 | 2010-06-21 | The purposes of the pancreatin zymolyte of bird's nest |
Country Status (5)
| Country | Link |
|---|---|
| JP (1) | JP5815202B2 (en) |
| KR (1) | KR20120106703A (en) |
| CN (2) | CN107049907A (en) |
| TW (1) | TW201105362A (en) |
| WO (1) | WO2010150737A1 (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109674731A (en) * | 2019-02-14 | 2019-04-26 | 厦门汇聚优品电子商务有限公司 | A kind of anti-apolexis composition containing roe essence and its application |
| CN109771320A (en) * | 2018-12-24 | 2019-05-21 | 佛山科学技术学院 | The skin care item and preparation method thereof of bird's nest peptide and plants essential oil compounding |
| CN113876612A (en) * | 2021-11-12 | 2022-01-04 | 湖北省麦诗特生物科技有限公司 | A facial cream composition containing liposome for increasing skin collagen density and its preparation method |
| CN114762698A (en) * | 2021-06-16 | 2022-07-19 | 艾琳娜医疗管理(南京)有限公司 | Nutrient solution for stimulating collagen repair of female face and equipment method thereof |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5815203B2 (en) * | 2009-06-23 | 2015-11-17 | コンビ株式会社 | Collagen production decrease inhibitor, external composition for skin containing the same, and cosmetics |
| JP5914577B2 (en) * | 2014-06-09 | 2016-05-11 | コンビ株式会社 | Collagen production decrease inhibitor, external composition for skin containing the same, and cosmetics |
| JP5914576B2 (en) * | 2014-06-09 | 2016-05-11 | コンビ株式会社 | Collagen production promoter, skin external composition containing the same, and cosmetics |
| KR102204367B1 (en) * | 2014-07-16 | 2021-01-19 | 주식회사 엘지생활건강 | Cosmetic compositions for improving skin wrinkle |
| JP7016117B2 (en) | 2016-01-18 | 2022-02-21 | コスメディ製薬株式会社 | Collagen cosmetics |
| TWI636794B (en) | 2017-11-06 | 2018-10-01 | 吳治增 | Nano composite material and application thereof |
| JP7285405B2 (en) * | 2020-02-18 | 2023-06-02 | 株式会社ファーマフーズ | Cosmetic composition derived from vitelline membrane and/or chalazae, and method for producing the same |
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- 2010-06-21 CN CN201610983140.4A patent/CN107049907A/en active Pending
- 2010-06-21 KR KR1020127001655A patent/KR20120106703A/en not_active Application Discontinuation
- 2010-06-21 CN CN2010800282277A patent/CN102458357A/en active Pending
- 2010-06-21 WO PCT/JP2010/060459 patent/WO2010150737A1/en active Application Filing
- 2010-06-23 TW TW099120421A patent/TW201105362A/en not_active IP Right Cessation
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| JPH09301819A (en) * | 1996-05-09 | 1997-11-25 | Nippon High Potsukusu:Kk | Cosmetic containing enka-extract |
| CN101084873A (en) * | 2007-06-19 | 2007-12-12 | 浙江大学 | Process for preparing edible bird's nest extraction used for cosmetics |
| CN101606900A (en) * | 2008-06-17 | 2009-12-23 | 深圳绵俪日用化工有限公司 | A kind of clean white beauty mask and processing method thereof that contains the Nidus collocaliae extracting solution |
| CN101757028A (en) * | 2008-12-22 | 2010-06-30 | 康贝株式会社 | Bone intensity improvement agent, skin collagen quantity improvement agent and oral composition and medicine part outer article containing at least one of the improvement agents |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109771320A (en) * | 2018-12-24 | 2019-05-21 | 佛山科学技术学院 | The skin care item and preparation method thereof of bird's nest peptide and plants essential oil compounding |
| CN109674731A (en) * | 2019-02-14 | 2019-04-26 | 厦门汇聚优品电子商务有限公司 | A kind of anti-apolexis composition containing roe essence and its application |
| CN114762698A (en) * | 2021-06-16 | 2022-07-19 | 艾琳娜医疗管理(南京)有限公司 | Nutrient solution for stimulating collagen repair of female face and equipment method thereof |
| CN113876612A (en) * | 2021-11-12 | 2022-01-04 | 湖北省麦诗特生物科技有限公司 | A facial cream composition containing liposome for increasing skin collagen density and its preparation method |
| CN113876612B (en) * | 2021-11-12 | 2023-09-22 | 湖北省麦诗特生物科技有限公司 | A facial cream composition containing liposome for increasing skin collagen density, and its preparation method |
Also Published As
| Publication number | Publication date |
|---|---|
| TWI559939B (en) | 2016-12-01 |
| WO2010150737A1 (en) | 2010-12-29 |
| KR20120106703A (en) | 2012-09-26 |
| JP5815202B2 (en) | 2015-11-17 |
| TW201105362A (en) | 2011-02-16 |
| CN107049907A (en) | 2017-08-18 |
| JP2011006331A (en) | 2011-01-13 |
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