CN102209551B - Composition for inhibiting fgf-5, hair tonic, and composition for external use on livestock - Google Patents

Composition for inhibiting fgf-5, hair tonic, and composition for external use on livestock Download PDF

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CN102209551B
CN102209551B CN2009801447876A CN200980144787A CN102209551B CN 102209551 B CN102209551 B CN 102209551B CN 2009801447876 A CN2009801447876 A CN 2009801447876A CN 200980144787 A CN200980144787 A CN 200980144787A CN 102209551 B CN102209551 B CN 102209551B
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团克昭
藤泽贤司
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Toyo Hakko Co Ltd
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Abstract

Provided is a composition for inhibiting FGF-5 having excellent inhibiting activity against FGF-5. This composition comprises at least one selected from rice bran/soy bean fermentation product; extract of flower petals of the genus Rosa belonging to the family Rosaceae; and iota-carrageenan. Also provided are a hair tonic and a composition for external use on livestock, both of which contain the composition for inhibiting FGF-5. As a result, the hair tonic and composition for external use on livestock are excellent in terms of hair growth.

Description

FGF-5 composite inhibiting, hair growth agent and livestock animals topical composition
Technical field
The present invention relates to be applicable to suppress compositions, hair growth agent (hair growth promoter, educate a mao agent) and the livestock animals topical composition of FGF-5.In more detail, the present invention relates to contain the FGF-5 composite inhibiting petal extract that is selected from Testa oryzae/soybean fermented product, Rosaceae gul and at least one in the IOTA type carrageenan, that suppress the inhibition superior activity of FGF-5, and contain this FGF-5 composite inhibiting, alopecia preventing effectiveness, piliation effect, support all excellent hair growth agent and livestock animals topical compositions of hair (hair maintenance) effect and long hair effect.
Background technology
Send out the reason of symptom as hypotrichosis and depilation, can enumerate anxiety, endocrine disturbance, heredity, poor circulation, malnutrition and aging etc.Send out symptom and recover piliation, foster hair and long hair in order to relax this hypotrichosis and depilation, people are studying various hair growth agents.
It is believed that the various materials such as hormone, cell growth factor, vitamin and medicine class are to the growth of hair and break up influential.Wherein, cell growth factor is considered to produce in the cells such as hair papilla and/or external root sheath, internal root sheath, by propagation, inhibition cell, can work to regulating growth cycle of hair (trophophase, catagen, resting stage).One of this cell growth factor is exactly FGF-5.Known this FGF-5 is the protein that is produced by the FGF-5 gene, has the character that suppresses hair follicle and hair growth.
Therefore, can think that inhibition FGF-5 is for relaxing hypotrichosis and a depilation symptom, recovering piliation, foster hair and long hair is effectively, and people have proposed to suppress the various schemes of the hair growth agent of FGF-5.For example, patent documentation 1 discloses a kind of hair growth agent that contains polypeptide, and this polypeptide contains a part of structure of FGF5, for FGF5 the inhibitory action of hair growth is had and suppresses active.In addition, patent documentation 2 discloses the hair growth promoter that contains Lac regis apis.In addition, the patent documentation 3 hair growth hair sealing compound having put down in writing testosterone-5α-reductase inhibitor and contained it.And, also put down in writing a kind of testosterone-5α-reductase inhibitor, it contains and is selected from bright red samguineus (Pycnoporus coccineus), bright red samguineus (Pycnoporus cinnabarinus), large spore DIHUA bacterium (Albatrellus confluens), ungulate cheese bacteria (Oligoporustephroleucus (Fr.) Gilbn.et Ryv), phellinus igniarius (Phellinusigniarius (Fr.) Qu é l.), Pleurotus citrinopileatus (Pleurotus Citrinopileatus Sing, the goldentop mushroom), HUAZIGU, JINZHENGU (plain gill fungus), at least one fruiting body extract in each sporophore of cherry rivet mushroom (Gomphidius roseus).
In addition, for the house pet beyond the people and livestock products animal too, for the depilation symptom that is caused by the external factor of dermatosis, aging and feeding environment etc. etc., also need to have the topical composition of hair growth effect.
The prior art document
Patent documentation
Patent documentation 1: JP 2002-293720 communique
Patent documentation 2: JP 2003-192541 communique
Patent documentation 3: Unexamined Patent 7-278006 communique
Summary of the invention
The problem that invention will solve
Yet, the hair growth agent of patent documentation 1 and patent documentation 2 records, its FGF-5 suppresses activity may not be abundant, in addition, and the hair growth agent of patent documentation 1～3 record, its hair growth effect can not be said satisfactory.
In addition, people also demand the depilation symptom of the livestock animals such as house pet and livestock products animal etc. is had the livestock animals topical composition of hair growth effect.And then, being used for the livestock products animals such as sheep of dress material etc. for its chaeta, people's demand can increase the livestock animals topical composition of this chaeta.
The object of the invention is to, provide the FGF-5 composite inhibiting of the inhibition excellence of FGF-5 and hair growth agent and the livestock animals topical composition of hair growth effect excellence.
Be used for solving the means of problem
The present invention as shown below.
1.FGF-5 composite inhibiting is characterized in that, contains the petal extract that is selected from Testa oryzae/soybean fermented product, Rosaceae gul and at least one in the IOTA type carrageenan.
2. above-mentioned 1 described FGF-5 composite inhibiting contains at least 2 kinds in the petal extract that is selected from Testa oryzae/soybean fermented product, Rosaceae gul and the IOTA type carrageenan.
3. above-mentioned 1 or 2 described FGF-5 composite inhibitings also contain the Pleurotus citrinopileatus extract.
4. hair growth agent is characterized in that, contains above-mentioned 1～3 each described FGF-5 composite inhibiting.
5. the livestock animals topical composition is characterized in that, contains above-mentioned 1～3 each described FGF-5 composite inhibiting.
The invention effect
FGF-5 composite inhibiting of the present invention is owing to contain the petal extract that is selected from Testa oryzae/soybean fermented product, Rosaceae gul and at least one in the IOTA type carrageenan, therefore have excellent FGF-5 and suppress active, thereby have the alopecia preventing effectiveness.
During in containing the petal extract that is selected from Testa oryzae/soybean fermented product, Rosaceae gul and IOTA type carrageenan at least 2 kinds of FGF-5 composite inhibiting of the present invention, have more excellent FGF-5 and suppress active, thereby have the alopecia preventing effectiveness.
In addition, FGF-5 composite inhibiting of the present invention has more excellent FGF-5 and suppresses active, thereby have the alopecia preventing effectiveness when further containing the Pleurotus citrinopileatus extract.
In addition, hair growth agent of the present invention has excellent piliation effect, supports hair effect and long hair effect by containing FGF-5 composite inhibiting of the present invention.
In addition, livestock animals of the present invention, has excellent piliation effect, supports hair effect and long hair effect livestock animals by containing FGF-5 composite inhibiting of the present invention with the external combination agent.
Description of drawings
Fig. 1 is the sensor curve figure by the BIACORE analyser of experimental example 1 acquisition.
Fig. 2 is the figure by the sensor of the BIACORE analyser of experimental example 2 acquisitions.
Fig. 3 is the figure by the sensor of the BIACORE analyser of experimental example 3 acquisitions.
Fig. 4 is the figure by the sensor of the BIACORE analyser of experimental example 4 acquisitions.
Fig. 5 is the figure by the sensor of the BIACORE analyser of experimental example 5 acquisitions.
Fig. 6 is visual by the mouse back through after 28 days of experimental example 6 acquisitions.
Fig. 7 is visual by the mouse back through after 28 days of experimental example 7 acquisitions.
Fig. 8 is visual by the mouse back through after 28 days of experimental example 8 acquisitions.
Fig. 9 is visual by the mouse back through after 28 days of experimental example 9 acquisitions.
Figure 10 is visual by the mouse back through after 28 days of experimental example 10 acquisitions.
Figure 11 is visual by the mouse back through after 28 days of experimental example 11 acquisitions.
Figure 12 is visual by the mouse back through after 28 days of experimental example 12 acquisitions.
Figure 13 is visual by the mouse back through after 28 days of experimental example 13 acquisitions.
Figure 14 is visual by the mouse back through after 28 days of experimental example 14 acquisitions.
Figure 15 is visual by the mouse back through after 28 days of experimental example 15 acquisitions.
Figure 16 is visual by the mouse back through after 28 days of experimental example 16 acquisitions.
Figure 17 is visual by the mouse back through after 28 days of experimental example 17 acquisitions.
Figure 18 is visual by the mouse back through after 28 days of experimental example 18 acquisitions.
The specific embodiment
In this manual, FGF-5 refers to the protein by the generation of FGF-5 gene.
[1] FGF-5 composite inhibiting
FGF-5 composite inhibiting of the present invention is characterised in that, contains the petal extract that is selected from Testa oryzae/soybean fermented product, Rosaceae gul and at least one in the IOTA type carrageenan.
FGF-5 composite inhibiting of the present invention contains the petal extract that is selected from Testa oryzae/soybean fermented product, Rosaceae gul and at least one in the IOTA type carrageenan, preferably contains at least 2 kinds in the petal extract that is selected from Testa oryzae/soybean fermented product, Rosaceae gul and the IOTA type carrageenan.
The petal extract of Testa oryzae/soybean fermented product, Rosaceae gul and IOTA type carrageenan have separately excellent FGF-5 and suppress active.Therefore, by containing at least one in them, can make the FGF-5 composite inhibiting that FGF-5 suppresses superior activity.
In addition, during in containing the petal extract that is selected from Testa oryzae/soybean fermented product, Rosaceae gul and IOTA type carrageenan at least 2 kinds, by by the synergy that becomes branch to produce more than 2 kinds, can make FGF-5 and suppress active more excellent FGF-5 composite inhibiting.
Above-mentioned Testa oryzae/soybean fermented product is by after using bacillus subtilis to make Testa oryzae and soybean peptide fermentation, carries out refinement treatment again and the fermented product made.
As above-mentioned Testa oryzae and above-mentioned soybean peptide, can use commercially available Testa oryzae and soybean peptide.Make Testa oryzae and the soybean peptide usage ratio in when fermentation, take Testa oryzae: the quality of soybean peptide is preferably (100: 1)～(100: 20) as benchmark.In the time of in being in this scope, be conducive to the breeding of bacterium most.
In addition, above-mentioned Testa oryzae/soybean fermented product also can by after using bacillus subtilis to make Testa oryzae extract and soybean extract fermentation, carry out refinement treatment and make again.In addition, as above-mentioned Testa oryzae extract and above-mentioned soybean extract, can use commercially available Testa oryzae extract and soybean extract.Make Testa oryzae extract and the soybean extract usage ratio in when fermentation, take the Testa oryzae extract: the quality of soybean extract is preferably (100: 1)～(100: 20) as benchmark.In the time of in being in this scope, be conducive to the breeding of bacterium most.
In addition, the bacillus subtilis (Bacillussubtilis) that is used for the fermentation of Testa oryzae and soybean peptide etc. is that representative aerobic spermatium forms bacterium.As this bacillus subtilis, so long as security performance accesses assurance, just there is not particular determination, consider preferred Bafillus natt (Bacillus natto) from the easiness that obtains, the viewpoint of manufacturing cost.
In order to utilize bacillus subtilis to make Testa oryzae and soybean peptide fermentation, obtain Testa oryzae/soybean fermented product, must give best pH value and nutritional condition etc.That is, need to add the enzyme such as the salts such as saccharide, sodium bicarbonate, the protease such as glucose and water (purifying waste water) etc.
The refinement treatment of carrying out after the fermentation in addition, is by the squeezing of the fermentation liquid after the above-mentioned fermentation, filtration are carried out.And then, as required, can be after fermentation, squeezing, filtering, then utilize the deodorization and discoloration of active carbon etc. to process and/or sedimentary Transformatin.In addition, after these are processed, also can again carry out filtration treatment.
In addition, as Testa oryzae/soybean fermented product, can use commercially available product.For example can enumerate " CELABIO " (Toyo Hakko Ltd.'s system) etc.
The petal extract of above-mentioned Rosaceae gul (being designated hereinafter simply as " Flos Rosae Multiflorae extract ") is the extract that extracts from the petal of Rosaceae gul.
As above-mentioned Rosaceae gul, so long as belong to the Flos Rosae Multiflorae (Rosa spp.) of Rosaceae Rosa, just there is not particular restriction.Can enumerate particularly french rose (Rosagallica), Rosa centifolia L (Rosa centifolia), muskrat (Rosa moschata), abnormal flavour Flos Rosae Multiflorae (Rosa foetida), huge colored Flos Rosae Multiflorae (Rosa gigantea), Flos rosae multiflorae (Rosamultiflora), wichuraiana (Rosa wichuraiana), Flos Rosae Rugosae (Rosa rugosa), the wild kinds such as Rosa Damascena (Rosa damascea), the garden-variety that perhaps their copulation is formed.
In addition, the above-mentioned Rosaceae gul that uses in order to obtain the Flos Rosae Multiflorae extract can directly use the part of its plant and/or all afterwards in results, re-uses after also can being dried.And then, when obtaining the extract of above-mentioned Rosaceae gul, extract raw material and can be do not pulverize or pulverize, also can remove the pretreatment such as impurity.
When obtaining the Flos Rosae Multiflorae extract, its extracting method, extraction conditions do not have particular determination.Use solvent as extracting, except water or hot water, can also use the mixed solvent of alcohols (methanol and ethanol etc.), ethyl acetate, normal hexane, acetone and other organic solvent or these organic solvents and water or hot water etc.Usually make the mixed solvent of water or hot water or water or hot water and alcohols.These extract with in the solvent, preferably use hot water.As mixed solvent, for example can enumerate Water-Methanol Mixtures, water-ethanol mixed solvent etc.In addition, when using the mixed solvent of above-mentioned water or hot water and alcohols, when take the total amount of mixed solvent during as 100 volume %, the ratio of alcohols is generally 20～90 volume %, is preferably 30～70 volume %, more preferably 40～60 volume %.
In addition, extraction during extraction is generally 3～7 with the pH value of solvent, is preferably 4～6, and more preferably 4～5.Extracting temperature does not have particular restriction, is preferably extract at room temperature or heating extraction.If extract at room temperature, for example, can be by raw material be impregnated in the Water-Methanol Mixtures equal solvent, and at room temperature place about 1～5 day and extract.When heating extraction, as heating-up temperature, be generally 40～100 ℃, be preferably 50～80 ℃, more preferably 50～70 ℃.By heating-up temperature is in this scope, can extract efficiently, be preferred therefore.
When using above-mentioned Flos Rosae Multiflorae extract in FGF-5 composite inhibiting of the present invention, the extract residue after usually will extracting (petal of the Rosaceae gul after the extraction) separates with extracting solution (containing Flos Rosae Multiflorae extract and the solution that extracts with solvent).The separation method of this moment does not have particular determination, can adopt the methods such as pressure filter and filtration (pressurization, normal pressure) to separate.The extracting solution of separating from the extract residue can be directly used in preparation FGF composite inhibiting of the present invention.In addition, extracting solution subsequently also can be further re-uses after with solvent removing wherein contained extraction.Removing the method for extracting with solvent from extracting solution does not have particular determination, for example can adopt, and spray drying method, evaporation and lyophilization etc. are carried out.The solid constituent of separated and collected from extracting solution (Flos Rosae Multiflorae extract solid composition) can directly use, and also can further make with extra care.That is, can filter by making after this solid constituent is dissolved in water and/or the organic solvent etc., remove field trash etc., desolventizing is made with extra care again.And then, can also implement as required sterilization treatment etc.
In addition, the Flos Rosae Multiflorae extract can use commercially available product.For example can enumerate " ROSE CYRSTA " (Toyo Hakko Ltd.'s system) etc.
Above-mentioned carrageenan is to extract from the red algae Sargassum and refining natural high molecular substance, is a kind of take molecular weight as 100,000～500, and 000 galactose and 3,6-Anhydrogalactose. are the polysaccharide of main constituent, have the sulfate group of half ester type in molecule.This carrageenan roughly is divided into 3 classes, KAPPA type, IOTA type and LAMBDA type.They can be distinguished according to the character of galactose repetitive contained in the carrageenan.Wherein, preferably contain IOTA type carrageenan among the present invention.
As the extraction raw material that is used for obtaining above-mentioned IOTA type carrageenan, can be red algae Sargassum etc., for example can enumerate the red algaes such as the Eucheuma spinosum of Eucheuma (Eucheuma spinosum).
In addition, when obtaining IOTA type carrageenan, its extracting method, extraction conditions do not have particular determination.As extracting method and extraction conditions, for example can enumerate following methods.With the Sargassum cold water washing, remove sand and other microgranules of existing in the Sargassum at first.At this, the carrageenan in the Sargassum is formed by constituent and the association of common cellulose of Sargassum, therefore, swelling can not occur in the process of cold water washing.Then, after cold water washing, then implement the hot water extraction operation.Then, this hot water extraction liquid is further at high temperature processed with aqueous alkali.As the alkali that uses in this aqueous alkali, can enumerate NaOH, Ca (OH) 2Or alkali or the alkaline earth metal hydroxide of KOH and so on.By the processing of this high temperature alkaline solution solution, can cause 3 in the galactose units, the generation of 6-dehydration bonding.Then, behind alkali modification, with extracting liquid filtering, remove the insoluble substances such as cellulose, hemicellulose and other microgranules, add subsequently acid, pH value is adjusted to below 9 or 9.In addition, after the extraction, can centrifugalize be set and utilize the bleaching process of bleach according to hope.Then, by in extracting solution, adding the alcohols such as KCl or isopropyl alcohol, it is precipitated from extracting solution as extract obtain.In addition, also can extracting solution be concentrated by making the extraction solvent evaporation of extracting solution, its drying is obtained.And then the dry thing that obtains by drying also can be pulverized with pulverizer etc., powdered.
Be explained, the solid constituent of separated and collected can directly use from extracting solution, also can further make with extra care.
In addition, IOTA type carrageenan can directly use commercially available product.For example can enumerate " VicarinSD389 " (FMC chemicals company limited system) etc.
In addition, FGF-5 composite inhibiting of the present invention can also contain the Pleurotus citrinopileatus extracting solution.
Above-mentioned Pleurotus citrinopileatus extract refers to the extract that extracts from sporophore of Pleurotus citrinopileatus etc.Above-mentioned Pleurotus citrinopileatus refers to the Pleurotus citrinopileatus (formal name used at school: " Pleurotus cornucopiae " (Pleurotus cornucopiae (paul.Pers.) Rolland)) of Basidiomycotina Eubasidiomycetes layer bacterium subclass Agaricales Pleurotaceae pleurotus.
In addition, as the composition that contains of Pleurotus citrinopileatus extract, can enumerate the glycosyl ceramides such as trehalose, mannitol, malic acid, glycogen, lysine, glutamic acid, D-xylose, ceramide, β-D glucan, ergosterol, unsaturated fatty acids, pyroglutamic acid, pentitol, hexitol, cystathionie, ornithine, glycosyl sphingolipid, ergothioneine,
Figure BDA0000060368290000081
(タ マ PVC ジ Application) etc.
As the Pleurotus citrinopileatus of Pleurotus citrinopileatus extract raw material, can utilize fresh sporophore or its dry thing, preferred fresh sporophore.When obtaining this Pleurotus citrinopileatus extract, its extracting method does not have particular determination.Use solvent as extracting, except water or hot water, can also use the mixed solvent of alcohols (methanol and ethanol etc.), ethyl acetate, normal hexane, acetone and other organic solvent or these organic solvents and water or hot water etc., usually make the mixed solvent of water or water and alcohols.In addition, the extraction during extraction is generally 5～9 with the pH value of solvent, is preferably 6～8.As extracting method, there is not particular determination, for example, raw material and solvent one to be reinstated pulverizer etc. pulverize, product are pulverized in preparation.Then, product be should pulverize and extract at room temperature or heating extraction carried out.If extract at room temperature was placed about 1 day～5 days.The situation of heating extraction as heating-up temperature, is generally 40～100 ℃, is preferably 70～90 ℃.By heating-up temperature is in this scope, can extract efficiently.
In addition, before product are pulverized in preparation, also can carry out pretreatment to raw material.As this pretreatment, at normal temperatures raw material is impregnated in the solvent, at room temperature placed 5 hours～3 days.And then, as required, in temperature chamber, the raw material behind the dipping is incubated (hatching).The condition of insulation does not have particular determination, is generally that temperature 60 C is above, humidity is more than 70%, and temperature retention time is 10 minutes～2 hours.Then, when carrying out above-mentioned pretreatment, use pretreated raw material, pulverize with pulverizer etc., product are pulverized in preparation.
When using the Pleurotus citrinopileatus extract in the FGF-5 composite inhibiting of the present invention, the extract residue (Pleurotus citrinopileatus after the extraction) after usually will extracting separates with extracting solution (the extraction solvent that contains the Pleurotus citrinopileatus extract).The separation method of this moment does not have particular determination, can adopt pressure filter and filtration (pressurization, normal pressure) etc. to separate.With the extracting solution after the extract residue separates, can be directly as the raw material of FGF-5 composite inhibiting of the present invention.In addition, extracting solution and then can also re-use after with solvent by removing wherein contained extraction.Removing the method for extracting with solvent from extracting solution does not have particular determination, for example can adopt, and spray drying method, evaporation and lyophilization etc. are carried out.The solid constituent of separated and collected can directly use from extracting solution, also can further make with extra care.That is, can filter making after this solid constituent is dissolved in water and/or the organic solvent etc., remove field trash etc., then desolventizing is made with extra care.And then, can also implement as required sterilization treatment etc.
FGF-5 composite inhibiting of the present invention contains at least one of selecting from Testa oryzae/soybean fermented product, Flos Rosae Multiflorae extract and I OTA type carrageenan, as mentioned above, preferably contain at least 2 kinds that from Testa oryzae/soybean fermented product, Flos Rosae Multiflorae extract and IOTA type carrageenan, select.
Testa oryzae/soybean fermented product, Flos Rosae Multiflorae extract and IOTA type carrageenan are all very high to the affinity of FGF-5, all have FGF-5 and suppress active.Therefore, FGF-5 composite inhibiting of the present invention has excellent inhibition activity for playing inhibiting FGF-5 to the growth of hair follicle and/or hair.Therefore, FGF-5 composite inhibiting of the present invention has the alopecia preventing effectiveness, and then, have the piliation effect, support hair effect and long hair effect.
As mentioned above, as containing at least 2 kinds the scheme of from Testa oryzae/soybean fermented product, Flos Rosae Multiflorae extract and IOTA type carrageenan, selecting, be illustrated in down.
(1) contains the compositions of Testa oryzae/soybean fermented product and Flos Rosae Multiflorae extract.
(2) contain the compositions of Testa oryzae/soybean fermented product and IOTA type carrageenan.
(3) contain the compositions of Flos Rosae Multiflorae extract and IOTA type carrageenan.
(4) contain the compositions of Testa oryzae/soybean fermented product, Flos Rosae Multiflorae extract and IOTA type carrageenan.
Among the present invention, more preferably contain at least one and the Pleurotus citrinopileatus extract from Testa oryzae/soybean fermented product, IOTA type carrageenan and Flos Rosae Multiflorae extract, selected, particularly preferably contain at least 2 kinds and Pleurotus citrinopileatus extract from Testa oryzae/soybean fermented product, IOTA type carrageenan and Flos Rosae Multiflorae extract, selecting.
In addition, the amount of contained Testa oryzae/soybean fermented product, Flos Rosae Multiflorae extract and IOTA type carrageenan does not have particular determination in the FGF-5 composite inhibiting of the present invention, can be only contain as the part of FGF-5 composite inhibiting, also can consist of the whole of FGF-5 composite inhibiting.Namely, when the gross mass with the FGF-5 composite inhibiting is set as 100 quality %, the total content of Testa oryzae/soybean fermented product, Flos Rosae Multiflorae extract and IOTA type carrageenan is 0.1～100 quality %, be preferably 3～100 quality %, more preferably 15～100 quality % are particularly preferably 20～100 quality %.When adding up to content in above-mentioned scope, can obtain enough FGF-5 inhibitions.
In addition, when FGF-5 composite inhibiting of the present invention contains the Pleurotus citrinopileatus extract, the content of Testa oryzae/soybean fermented product, Flos Rosae Multiflorae extract and IOTA type carrageenan and Pleurotus citrinopileatus extract does not have particular determination, can be used as the part of FGF-5 composite inhibiting and contain, also can consist of the whole of FGF-5 composite inhibiting.Namely, when take the gross mass of FGF-5 composite inhibiting during as 100 quality %, the total content of Testa oryzae/soybean fermented product, Flos Rosae Multiflorae extract and IOTA type carrageenan and Pleurotus citrinopileatus extract is 0.1～100 quality %, be preferably 3～100 quality %, more preferably 15～100 quality % are particularly preferably 20～100 quality %.When adding up to content in above-mentioned scope, can obtain enough FGF-5 inhibitions.
The solid component concentration of FGF-5 composite inhibiting does not have particular determination, is generally 0.0005～100 quality %, is preferably 0.001～50 quality %, and more preferably 0.01～5 quality % is particularly preferably 0.01～1.5 quality %.
Be explained, the solid constituent in the solid component concentration of FGF-5 composite inhibiting refers to the solid constituent that comprises Testa oryzae/soybean fermented product, Flos Rosae Multiflorae extract and IOTA type carrageenan contained in the constituent.
In addition, when the FGF-5 composite inhibiting contained the Pleurotus citrinopileatus extract, the solid constituent in the solid component concentration referred to the solid constituent that comprises Testa oryzae/soybean fermented product, Flos Rosae Multiflorae extract and IOTA type carrageenan and Pleurotus citrinopileatus extract as constituent.
Contain at least 2 kinds the situation of from Testa oryzae/soybean fermented product, IOTA type carrageenan and Flos Rosae Multiflorae extract, selecting at the FGF-5 composite inhibiting, when the total solid constituent with Testa oryzae/soybean fermented product, IOTA type carrageenan and Flos Rosae Multiflorae extract was set as 100 quality %, the mass ratio of the side in these 2 kinds of materials and the opposing party's solid constituent can be 10～90 quality % and 10～90 quality %.
In addition, when the FGF-5 composite inhibiting contains these 3 kinds of Testa oryzae/soybean fermented product, IOTA type carrageenan and Flos Rosae Multiflorae extracts, when the total solid constituent with Testa oryzae/soybean fermented product, IOTA type carrageenan and Flos Rosae Multiflorae extract is set as 100 quality %, the mass ratio of these 3 kinds of solid constituents, Testa oryzae/soybean fermented product can be 1～90 quality %, IOTA type carrageenan can be 1～90 quality %, and the Flos Rosae Multiflorae extract can be 1～90 quality %.
In addition, when the FGF-5 composite inhibiting contains the Pleurotus citrinopileatus extract, the mass ratio of the total amount of at least one of from Testa oryzae/soybean fermented product, IOTA type carrageenan and Flos Rosae Multiflorae extract, selecting and the solid constituent of Pleurotus citrinopileatus extract, when above-mentioned solid constituent is set as 100 quality %, can be 10～90 quality % and 10～90 quality %.
Be explained, solid component concentration refers to adopt the quality % (lower with) of the drying solid composition that general loss on drying (lower dry 4 hours at 105 ℃) records.In addition, concentrated referring under the state that material is remained on the moisture evaporation is to improve the processing of solid component concentration.Concentrated method does not have particular determination, for example can enumerate, and concentrating under reduced pressure and instant decompression are concentrated etc.
Form for FGF-5 composite inhibiting of the present invention does not have particular determination.Usually use with the various forms of fermented product and extract.For example, can make graininess after liquid state, solid-state, Powdered, graininess, the granulation etc.
In addition, containing fermented product (the cultivation and fermentation liquid that cultivation obtains) and the extracting solution of composition, can be the stock solution after filtering, and also can be the liquid to post processings such as it decolour, perhaps also can be the concentrated solution after it is concentrated.In addition, also can be to adopt the known method such as lyophilizing to remove solid content or the powdery powder of solvent.
In addition, FGF-5 composite inhibiting of the present invention is except containing Testa oryzae/soybean fermented product, Flos Rosae Multiflorae extract and IOTA type carrageenan and Pleurotus citrinopileatus extract, and the FGF-5 that can also contain other suppresses composition.FGF-5 as other suppresses composition, can enumerate lambda carrageenan, KAPPA type carrageenan etc.
FGF-5 composite inhibiting of the present invention has FGF-5 and suppresses active.This FGF-5 suppresses active assay method, for example can adopt the BIACORE analyser to measure.
The mensuration that adopts this BIACORE analyser be adopt the BIACORE analytic process measure FGF-5 to be measured suppress active composition (hereinafter referred to as " sample ") and the FGF-5 protein that is immobilized occur in conjunction with the time in conjunction with rate constant and the rate constant that dissociates, derive thus that the KD value carries out.FGF-5 is fixed on the surface of sensor chip of 2 intermolecular interaction determinators, adopts surface plasma body resonant vibration method (using 2 intermolecular interaction determinators etc.) to measure combination with sample, measure thus the affinity with FGF-5.
Particularly, after making the lip-deep dextran activation of sensor chip (" CM5 " BIACORE company system) of the 2 intermolecular interaction determinators (BIACORE company system) (being designated hereinafter simply as " determinator ") that are coated on based on the surface plasma body resonant vibration effect, make the dextran of this activation and a certain amount of FGF-5 carry out the amine bonding.This sensor chip is installed in the determinator, in order to observe the interaction between FGF-5 and the sample, makes the serial dilution solution of sample pass through according to priority minim channel.When occuring to interact between FGF-5 and the sample, in the lines rising of sensor curve figure expression interaction (combination).In addition, when losing above-mentioned interaction, demonstrate immediately the pattern of lines fast-descending.In addition, even owing to above-mentioned interactional reaction causes interactant residual, also can make FGF-5 return to immobilized initial conditions by regenerative operation, thereby can repeat to measure.As this regenerative operation, as long as use 30 seconds of sodium hydroxide effect of 8mmol/l.
The research of affinity is obtained binding constant and the constant that dissociates based on the response speed opinion by the slope (rate of change in the certain hour) of this sensor curve figure, and then, calculate reaction constant KD (M).The value of this reaction constant is less, can judge that 2 intermolecular affinitys are higher.In addition, the value of this reaction constant KD for general antigen antibody reaction etc., is 10 -7～10 -8M for the high material of the specificity of biotin and avidin and so on, is 10 -12～10 -13M.For example, when reaction constant KD be 10 -8During M, with respect to 10 -7M, it is high 10 times to be evaluated as affinity.And its affinity is higher, and expression is stronger with the interaction of FGF-5, and " sample-(FGF-5) complex " that formed by sample and FGF-5 becomes easier and form, thereby can improve FGF-5 inhibition activity.
In addition, per 1 day use amount of FGF-5 composite inhibiting of the present invention does not have particular determination, and the total amount of above-mentioned essential component is pressed solid constituent and converted, and is preferably 0.0005～50g, more preferably 0.001～5g, more preferably 0.01～1.5g.When the consumption when 1 day is in the above-mentioned scope, has excellent alopecia preventing effectiveness.
In addition, suppress active FGF-5 composite inhibiting of the present invention and be used for external preparation by having FGF-5, can make and have the alopecia preventing effectiveness, the piliation effect, support the external preparation of hair effect and long hair effect.
[2] hair growth agent
Hair growth agent of the present invention is characterised in that, contains above-mentioned FGF-5 composite inhibiting.This FGF-5 compositions as mentioned above.
The content of contained above-mentioned FGF-5 composite inhibiting does not have particular determination in the hair growth agent of the present invention, can be used as the part of hair growth agent and contains, and can consist of the whole of hair growth agent yet.When consisting of hair growth agent whole, the FGF-5 composite inhibiting directly can be used as hair growth agent.
In addition, when the gross mass with hair growth agent was set as 100 quality %, the content of FGF-5 composite inhibiting was 0.1～100 quality %, was preferably 3～100 quality %, and more preferably 15～100 quality % are particularly preferably 20～100 quality %.When the content of compositions is in the above-mentioned scope, has excellent hair growth effect.
The solid component concentration of hair growth agent of the present invention does not have particular determination, is generally 0.0005～100%, is preferably 0.001～50%, and more preferably 0.01～5%, be particularly preferably 0.01～3%.
Be explained, said solid constituent in the solid component concentration of above-mentioned hair growth agent, refer to as the constituent of FGF-5 composite inhibiting, comprise the solid constituent of Testa oryzae/soybean fermented product, Flos Rosae Multiflorae extract and I OTA type carrageenan and the solid constituent that comprises Testa oryzae/soybean fermented product, Flos Rosae Multiflorae extract, IOTA type carrageenan and Pleurotus citrinopileatus extract.
In addition, the FGF-5 composite inhibiting as contained in the hair growth agent of the present invention is preferably the compositions that contains the Pleurotus citrinopileatus extract.Because the Pleurotus citrinopileatus extract has more excellent piliation effect and long hair effect, therefore, contain the FGF-5 composite inhibiting of Pleurotus citrinopileatus extract by use, can make piliation effect and the more excellent hair growth agent of long hair effect.
Preferred version as hair growth agent of the present invention is illustrated in down.
(1) comprises the hair growth agent of the FGF-5 composite inhibiting that contains Testa oryzae/soybean fermented product and Pleurotus citrinopileatus extract.
(2) comprise the hair growth agent of the FGF-5 composite inhibiting that contains Flos Rosae Multiflorae extract and Pleurotus citrinopileatus extract.
(3) comprise the hair growth agent of the FGF-5 composite inhibiting that contains IOTA type carrageenan and Pleurotus citrinopileatus extract.
(4) comprise the hair growth agent of the FGF-5 composite inhibiting that contains Testa oryzae/soybean fermented product and IOTA type carrageenan and Pleurotus citrinopileatus extract.
(5) comprise the hair growth agent of the FGF-5 composite inhibiting that contains Testa oryzae/soybean fermented product and Flos Rosae Multiflorae extract and Pleurotus citrinopileatus extract.
(6) comprise the hair growth agent of the FGF-5 composite inhibiting that contains IOTA type carrageenan and Flos Rosae Multiflorae extract and Pleurotus citrinopileatus extract.
(7) comprise the hair growth agent of the FGF-5 composite inhibiting that contains Testa oryzae/soybean fermented product, IOTA type carrageenan and Flos Rosae Multiflorae extract and Pleurotus citrinopileatus extract.
In the such scheme, because the Pleurotus citrinopileatus extract has more excellent piliation effect and long hair effect, IOTA type carrageenan has more excellent FGF-5 inhibitory action, and therefore, the hair growth agent that contains (3)～(7) of Pleurotus citrinopileatus extract and IOTA type carrageenan is preferred.And then, particularly preferably be (6) and (7), at least one that contained FGF-5 composite inhibiting contains Pleurotus citrinopileatus extract and IOTA type carrageenan and selects from Testa oryzae/soybean fermented product and Flos Rosae Multiflorae extract in their hair growth agent.If use the FGF-5 composite inhibiting of at least one contain Pleurotus citrinopileatus extract and IOTA type carrageenan and from Testa oryzae/soybean fermented product and Flos Rosae Multiflorae extract, to select, then FGF-5 inhibition and the hair growth effect that they have can be brought into play synergistically, thereby piliation effect and the more excellent hair growth agent of long hair effect can be made.
In addition, can contain known additive in the hair growth agent of the present invention.For example can enumerate cell vitalizing agent, vitamins, hormones, vasodilation, amino acids, anti-inflammatory agent, antibacterial, keratin-lytic agent etc.They can be used alone or in combination of two or more kinds.
Particularly, as above-mentioned cell vitalizing agent, can enumerate pentadecanoic acid glyceride, Hinokitiol, forskolin, trans-3,4 '-dimethyl-3-hydroxyl flavanone etc.
As the said vitamin class, can enumerate vitamin A, vitamin B complex, tocopherols, nicotinic acid, nicotiamide, palmitic retinol, beta-carotene, calciferol, folic acid, biotin, D-panthenol, acetyl pantoyl ethylether, pantothenic acid class etc.
As above-mentioned hormones, can enumerate estradiol etc.
As above-mentioned vasodilation, can enumerate minoxidil, when medicament extracting solution etc.
As above-mentioned amino acids, can enumerate arginine, aspartic acid, methionine, serine, glycine, glutamic acid, cystine etc.; As antiinflammatory, can enumerate β-enoxolone, Radix Glycyrrhizae acids, pyridoxine hydrochloride etc.
As above-mentioned antibacterial, can enumerate isopropyl methyl phenol, piroctone olamine etc.
Hair growth agent of the present invention contains above-mentioned FGF-5 composite inhibiting of the present invention etc., and only otherwise suppress action effect of the present invention, can also contain other composition.For example, can add the known composition that contains in the past cosmetics and the hair growth agent etc.Particularly, can enumerate surfactant, oil preparation, alcohol, pH adjusting agent, antiseptic, antioxidant, viscosifier, pigment, spice etc.They can be used alone or in combination of two or more kinds.
The form of hair growth agent of the present invention does not have particular determination, can suit as required to select various forms.For example, aqueous (astringent, emulsion, dispersion liquid etc.), gel, emulsus, spray form, Powdered etc.In aqueous situation, can be aqueous by what obtain in water solvents such as mentioned component etc. is water-soluble, ethanol, propylene glycol etc., in addition, can also by hydrocarbon ils, olive oil, Semen Tritici aestivi germ oil, macadamia nut oil, Semen Maydis oil, Testa oryzae oil, rice germ oil, adlay oil, Jojoba oil and the animal oil such as Semen Vitis viniferae wet goods vegetable oil, squalane and house oil such as suitable fluid,matching paraffin body, other oil and the rudimentary ethanol such as ゲ Le マ one Le modification ethanol in a kind or 2 kinds with on obtain.In addition, in gelatinous situation, can be by except cooperating above-mentioned oil or ethanol, also cooperate in the viscosifier such as cellulose derivative, PVP and carboxy vinyl polymer such as CMC a kind or 2 kinds with on make.And then, in the situation of emulsus, can only be the oil components such as liquid paraffin, vaseline, Cera Flava, also can be the emulsion that these oil components is formed with water emulsifying with surfactant.In addition, as the emulsifying type, can be in W/O oil-in-water type, the O/W oil-in-water type any.
In addition, in the situation of spray form, except mentioned component, can also cooperate the lower alcohols such as normal propyl alcohol, isopropyl alcohol; The imflammable gas such as butane, propane, iso-butane, liquefied petroleum gas, dimethyl ether; The compression such as nitrogen, oxygen, carbon dioxide, nitrous oxide gas gas also can use their mixture.In this case, dimethyl ether can be cooperated, liquefied petroleum gas is cooperated, liquefied petroleum gas is cooperated to obtain in (liquefied petroleum gas/nitrogen)=(99.99/0.01)～(50/50) with nitrogen in mass ratio in (liquefied petroleum gas/dimethyl ether)=(99/1)～(1/99) in mass ratio with dimethyl ether in (dimethyl ether/nitrogen)=(99.99/0.01)～(50/50) in mass ratio with nitrogen.
In addition, per 1 day use amount of above-mentioned hair growth agent does not have particular determination, and the above-mentioned total amount that contains composition converts by solid constituent, is preferably 0.0005～50g, more preferably 0.001～5g, more preferably 0.01～3g.In the time of in per 1 day use amount is in above-mentioned scope, has excellent alopecia preventing effectiveness.
[3] livestock animals topical composition
Livestock animals of the present invention is characterised in that with topical composition and contains above-mentioned FGF-5 composite inhibiting.
Livestock animals topical composition of the present invention as long as contain above-mentioned FGF-5 composite inhibiting, does not just have particular determination, more preferably comprises the FGF-5 composite inhibiting that contains the Pleurotus citrinopileatus extract.
Livestock animals of the present invention is with the purpose of external preparation, and hair growth agent of the present invention is used for people animal in addition, is as using the topical composition of object take livestock animals such as house pet and livestock products animals.Therefore, livestock animals of the present invention is with in the topical composition, contain composition, contain the manufacture method of composition, the character that contains composition, the physical property that contains composition, other forms that contain composition and this topical composition etc., with hair growth agent of the present invention be same.
Use livestock animals of the present invention not have particular determination with the livestock animals of topical composition.This livestock animals refers to the birds and beasts of artificial rearing, for example can enumerate the mammalss such as Canis familiaris L., cat, horse, sheep, goat, cattle, pig, donkey, camel, alpaca, Cavia porcellus, rabbit, mink, rat and mouse; The birdss such as chicken, duck, goose, turkey, pheasant and Carnis Coturnicis japonicae.
Livestock animals of the present invention, therefore has piliation, long hair to animals such as livestock animals and prevents the effect of hair anticreep owing to contain above-mentioned composition with topical composition.As the operational version of livestock animals of the present invention with topical composition, for example can enumerate following scheme.
(1) for increasing the external preparation of the harvest yield of the chaeta that obtains from the livestock products animal.
(2) for the external preparation depilation that causes because of dermatosis and aging etc. etc., that be used for setation, become mildewed or prevent to lose hair or feathers.
Scheme as above-mentioned (1), be used for the livestock products animals such as sheep of dress material etc. for chaeta, can make the growth rate that can promote its chaeta and chaeta amount increase, and effectively increase the livestock animals topical composition of the chaeta receipts amount that obtains from these livestock products animals.
As the scheme of above-mentioned (2), can make the livestock animals that causes for the strain that produces because of dermatosis, aging and feeding environment etc. the depilation symptom, be used for setation, become mildewed or prevent the livestock animals external preparation that loses hair or feathers.
Livestock animals of the present invention with topical composition with the extract that from natural goods, extracts as main component, and compare as the external preparation of main component with synthetic ingredient, can more safely use livestock animals.Especially, for livestock animals, when at skin coating topical composition, animal tends to lick the topical composition at this coating position.And livestock animals of the present invention is with topical composition because take the extract that extracted by natural goods as main constituent, therefore, even absorbed also and can use safely by livestock animals.
In addition, livestock animals of the present invention with topical composition for livestock animals have the setation effect, the effect that becomes mildewed or anti-hair removal effect, therefore, suitable hair growth agent, long hair agent or the lint shedding resisting agent for clothes etc. of using as livestock animals.
Above-mentioned livestock animals does not have particular determination with per 1 day use amount of topical composition, and the above-mentioned total amount that contains composition converts by solid constituent, is preferably 0.0005～50g, more preferably 0.001～5g, more preferably 0.01～3g.In the time of in per 1 day use amount is in above-mentioned scope, can makes and have excellent anti-hair removal effect and the livestock animals topical composition of setation effect and the effect that becomes mildewed.
Embodiment
Below enumerate embodiment, illustrate in greater detail the present invention, but in the scope that does not exceed purport of the present invention, the present invention is not limited to these embodiment.In addition, the extracting solution of various extracts (extracting solution) for adopting water-medium to obtain for IOTA type carrageenan and lambda carrageenan, uses its dried powder.
Be explained, hereinafter, unless otherwise specified, % is quality criteria.
[1] preparation of the material composition of FGF-5 composite inhibiting
Adopt following method to prepare Flos Rosae Multiflorae extract, Testa oryzae/soybean fermented product and Pleurotus citrinopileatus extract.In addition, IOTA type carrageenan and lambda carrageenan use following commercially available product.
(1) preparation example 1: the preparation of Flos Rosae Multiflorae extract 1
As the Rosaceae gul, use Rosa centifolia L (Rosa centifolia).After the dry thing of its petal (pulverizing product) 20g extracted 30 minutes with 70 ℃ hot water 500ml, filtration (the use perlite is as filtration adjuvant).Then, behind the concentrating under reduced pressure, lyophilization obtains Flos Rosae Multiflorae extract 10g.Then, with Flos Rosae Multiflorae extract dilute with water obtained above, be adjusted to solid component concentration 2% rear use.
(2) preparation example 2: the preparation of Testa oryzae/soybean fermented product 1
After Testa oryzae 30g, disodium-hydrogen 10g, soybean peptide 4g, sodium bicarbonate 45g, glucose 5g, alkaline protease 0.1g, water 500g mixing, be adjusted to 8.8～9.0 culture medium with pH value at 42 ℃ of lower bacillus nattos (Bacillus natto) of cultivating after 48 hours, after this culture fluid squeezing, filtration, using the activated carbon decolorizing deodorize, again filter, obtain aqueous Testa oryzae/soybean fermented product 5g.Then, with Testa oryzae obtained above/soybean fermented product dilute with water, solid component concentration is adjusted to 0.8% rear use.
(3) preparation example 3: the preparation of Pleurotus citrinopileatus extract
The Pleurotus citrinopileatus 1Kg of aquatic foods flooded 20 hours in 25L water after, be that about 85 ℃, humidity are in about 90% the temperature chamber stifling (steam) at design temperature.Then, to all be merged together through the water that uses in stifling Pleurotus citrinopileatus, the stifling operation and with the composition that steam extracts from Pleurotus citrinopileatus, in Temperature Setting is 40 ℃ water-bath, be transferred in the pulverizer (" Port one タ Block Le ミ キ サ one A510 " TERAOKA company system).This is pulverized and uses mixer, carries out with the speed about 1200rpm, until become pasty state.Then, with the infusion under about 85～90 ℃ condition of this thickener.Then, with boil extracting solution all filter, remove solid constituent.With the filtrate that the obtains sterilization of heating, obtain to contain the Pleurotus citrinopileatus extract and extract extracting solution 20L with solvent (water).In addition, the sample that this extracting solution is diluted with water to 10mg/ml under the absorbance of 280nm as sample 1, be diluted to the sample of 20mg/ml as sample 2.Be explained, the solid component concentration of the Pleurotus citrinopileatus extract of this sample 1 is 1%, and the solid component concentration of the Pleurotus citrinopileatus extract of sample 2 is 2%.
(4) IOTA type carrageenan
" Vicarin SD 389 " (the FMC chemicals company limited system) of use.
(5) lambda carrageenan
Commodity in use name " Viscarin GP209 " (FMC chemicals company limited system).
[2] FGF-5 inhibition experiment (BIACORE analyser)
Adopt following method, measure the material composition of FGF-5 composite inhibiting obtained above to the affinity of FGF-5, investigate thus the FGF-5 inhibition.
Make the 2 intermolecular interaction determinators (" BIACORE 2000 " that are coated in based on surface plasma (プ ラ ズ モ Application) resonance effect, GE healthcare life science company system) sensor chip (" CM5 " of (being designated hereinafter simply as " determinator "), GEhealthcare life science company system) after lip-deep dextran (Sensor Chip CM 5) activation, make dextran and a certain amount of FGF-5 protein of this activation carry out the amine bonding, this sensor chip is installed in the determinator, in order to observe FGF-5 protein and sample (the FGF-5 composite inhibiting that is called experimental example 1～5, interaction down together) is flow through minim channel the serial dilution solution of sample is in turn worked.
The said determination experiment is carried out in accordance with the following methods.
For FGF-5 being fixed in 4 flow cells on the Biacore sensor chip CM5 (having applied the chip of Sensor Chip CM 5), in Sensor Chip CM 5, carry out 18 minutes amine coupling processing.Then, by FGF-5 was injected 300 seconds with the flow velocity that 10 μ l/ divide, FGF-5 is fixed on the sensor chip.In addition, the free amine group and the ethanolamine 1mol that exist on the chip surface were reacted 18 minutes, not make it non-specific binding occurs.
Then, when research interacts, make the HBS buffer agent (NaCl of the HEPES of 10mmol/l, pH7.4,150mmol/l, the EDTA of 3mmol/l) as buffer flow through 150 seconds (10 μ l/ divide).Then, the flow velocity that divides with 10 μ l/ injected sample 180 seconds, obtained to demonstrate the interactional sensor curve figure between FGF-5 and the sample.
In addition, behind the sample determination, make the sodium hydroxide solution of 8mmol/l flow through 30 seconds, make flow cell regeneration with the pulse of pH about 8.0.
In addition, the sensor curve figure that KD (M) value is obtained by the sample of measuring each 8 kinds of variable concentrations, (BI Aevaluation ver.3.0) calculates with BIACORE dedicated analysis software.
Use the material composition of the FGF-5 composite inhibiting that following experimental example 1～5 obtains, the preparation sample is measured KD (M) value.Then, KD (M) value that obtains is shown in table 1.In addition, the sensor curve figure with following experimental example 1～5 is shown in Fig. 1～5.Be explained, among the sensor curve figure of Fig. 1～5, each 8 kinds of curve chart is by the tactic curve chart of high step-down according to concentration from top to bottom.
In addition, preparation method and the concentration of the test sample in the experimental example 1～5 are as described below.Be explained, as the solvent for the preparation of (dilution) following each sample, use above-mentioned HBS buffer agent.
(1) experimental example 1
The Flos Rosae Multiflorae extract that preparation example 1 is obtained dilutes with above-mentioned HBS buffer agent, and the making solid component concentration is respectively the sample of 8 kinds of concentration of 0.002%, 0.001%, 0.0005%, 0.00025%, 0.000125%, 0.0000625%, 0.0000313% and 0.0000156%.Then, use these samples, measure the affinity to FGF-5.
(2) experimental example 2
Testa oryzae/soybean fermented product that preparation example 2 is obtained dilutes with above-mentioned HBS buffer agent, and the making solid component concentration is respectively the sample of 8 kinds of concentration of 0.0008%, 0.0004%, 0.0002%, 0.0001%, 0.00005%, 0.000025%, 0.0000125% and 0.00000625%.Then, use these samples, measure the affinity to FGF-5.
(3) experimental example 3
Above-mentioned IOTA type carrageenan is diluted with above-mentioned HBS buffer agent, make the sample that solid component concentration is respectively 8 kinds of concentration of 1%, 0.5%, 0.25%, 0.125%, 0.0625%, 0.00313%, 0.00156% and 0.000781%.Then, use these samples, measure the affinity to FGF-5.
(4) experimental example 4
The Pleurotus citrinopileatus extract (sample 1) that preparation example 3 is obtained dilutes with above-mentioned HBS buffer agent, and the making solid component concentration is respectively the sample of 8 kinds of concentration of 1%, 0.5%, 0.25%, 0.125%, 0.0625%, 0.00313%, 0.00156% and 0.000781%.Then, use these samples, measure the affinity to FGF-5.
(5) experimental example 5
Above-mentioned lambda carrageenan is diluted with above-mentioned HBS buffer agent, make the sample that solid component concentration is respectively 8 kinds of concentration of 1%, 0.5%, 0.25%, 0.125%, 0.0625%, 0.00313%, 0.00156% and 0.000781%.Then, use these samples, measure the affinity to FGF-5.
[table 1]
Sample KD (M) value
Experimental example 1 The Flos Rosae Multiflorae extract 1.60×10 -6
Experimental example 2 Testa oryzae/soybean fermented product 6.04×10 -6
Experimental example 3 IOTA type carrageenan 2.95×10 -8
Experimental example 4 The Pleurotus citrinopileatus extract 2.18×10 -5
Experimental example 5 Lambda carrageenan 1.07×10 -6
In addition, diluted concentration about above-mentioned sample, according to each the FGF-5 composite inhibiting that makes experimental example 1～5 from 100 μ mol/l concentration, respectively by 1/2 stepwise dilution, become 8 kinds of concentration of 50 μ mol/l, 25 μ mol/l, 12.5 μ mol/l, 6.25 μ mol/l, 3.125 μ mol/l, 1.56 μ mol/l and 0.78 μ mol/l.
Be explained, in the calculating of above-mentioned concentration, suppose that the molecular weight that contains composition contained in Flos Rosae Multiflorae extract and the Testa oryzae/soybean fermented product is 10,000, and, suppose that the molecular weight that contains composition, IOTA type carrageenan and lambda carrageenan contained in the Pleurotus citrinopileatus extract is 100,000.
[3] preparation and the evaluation of hair growth agent (livestock products animal topical composition)
Experimental example 6～18 (use C3H in 6 age in week is the piliation experiment of male mice)
(1) evaluation methodology of hair growth agent (livestock products animal topical composition)
With 65 6 age in week C3H be that male mice (the three ラ ボ サ of association, one PVC ス Co., Ltd. system) is anaesthetized with pentobarbital, shave off with the chaeta (rectangle of 2cm * 4cm) of clippers with above-mentioned mouse back, this rectangular segment of having shaved hair is shaved light with razor again, obtain laboratory mice.
65 laboratory mices obtained above are divided into 13 groups, every group each 5, should be used for experimental example 6～18 by each 5 mice respectively.
Then, per 1 day each 20 μ l (to per 1) of hair growth agent (livestock products animal topical composition) with the experimental example 6～18 shown in following, to laboratory mice (each experimental example is 5) 1 day 1 time, be coated on the back central part of shaving the light hair of laboratory mice.Then, this operation was carried out 28 days, measured the hair growth effect of the chaeta of mice.
The mensuration of above-mentioned hair growth effect, the laboratory mice that uses will each experimental example begin by 28 days from experiment after is anaesthetized with pentobarbital, 5 mice set with each experimental example, under certain distance (from the distance of the estrade 25cm that places mice), certain brightness (under the same fluorescent lamp), take image with digital camera.Then, with the mouse back chaeta do not shave light common 10 age in week C3H be that the chaeta state image at male mice back compares.Then, with the chaeta of the mice of using in the experiment clearly the area of few part quantize, with the meansigma methods of the area that quantizes in 5 mices of each experimental group as without gross area.Carry out statistical disposition with what this quantized without gross area, measure thus hair growth effect.Be explained, quantize and statistical disposition use analysis software (" Quantity One " BIO RAD company system).In addition, with experimental example 6～18 without gross area (mm 2) and the poor (Δ without gross area without gross area and experimental example 6 of experimental example 7～18 1Value) is shown in table 2.And the mouse back image after experimental example 6～18 is obtained 28 days is shown in Fig. 6～18.
(2) preparation of hair growth agent (livestock products animal topical composition)
In the preparation process of the hair growth agent (livestock products animal topical composition) shown in following, for Flos Rosae Multiflorae extract and Testa oryzae/soybean fermented product, use the FGF-5 composite inhibiting of being made by following sample 3 and 4.
(1) Flos Rosae Multiflorae extract 2 (sample 3)
With trade name " ROSE CYRSTA " (Toyo Hakko Ltd.'s system) as FGF-5 composite inhibiting (sample 3).The solid component concentration of this Flos Rosae Multiflorae extract is 4%.
(2) Testa oryzae/soybean fermented product 2 (sample 4)
With trade name " CELABIO " (Toyo Hakko Ltd.'s system) as FGF-5 composite inhibiting (sample 4).The solid component concentration of this Testa oryzae/soybean fermented product is 0.8%.
Below the hair growth agent (livestock products animal topical composition) that uses in the experimental example 6～18 is shown in.
(1) experimental example 6
Ethanol 200 μ l and the water 200 μ l of purity 99.5% are joined in the Eppendorf tube of capacity 1.5ml, with its mixing, make about 50% alcoholic solution.
(2) experimental example 7
To adding 1% minoxidil in about 50% alcoholic solution that obtains with experimental example 6 same operations, making gross mass is 100%, obtains minoxidil and contains thing.
(3) experimental example 8
The ethanol 200 μ l of purity 99.5% and Pleurotus citrinopileatus extract (solid component concentration 2%) the 200 μ l of above-mentioned experimental example 4 (sample 2) acquisition are joined in the Eppendorf tube of capacity 1.5ml, with its mixing, obtain the Pleurotus citrinopileatus extract and contain thing (solid component concentration 1%).
(4) experimental example 9
To operating equally the IOTA type carrageenans that add 0.5% experimental example 3 in about 50% alcoholic solution that obtains with experimental example 6, making gross mass is 100%, obtains IOTA type carrageenan and contains thing.
(5) experimental example 10
The ethanol 200 μ l of purity 99.5% and Testa oryzae/soybean fermented product (solid component concentration 0.8%) 200 μ l of said sample 4 are joined in the Eppendorf tube of capacity 1.5ml, with its mixing, obtain Testa oryzae/soybean fermented product and contain thing (solid component concentration 0.4%).
(6) experimental example 11
The ethanol 200 μ l of purity 99.5% and Flos Rosae Multiflorae extract (solid component concentration 4%) the 200 μ l of said sample 3 are joined in the Eppendorf tube of capacity 1.5ml, with its mixing, obtain the Flos Rosae Multiflorae extract and contain thing (solid component concentration 2%).
(7) experimental example 12
To contain with the Pleurotus citrinopileatus extract that the same operation of experimental example 8 obtains thing (solid component concentration 1%) 400 μ l and contain thing 400 μ l with IOTA type carrageenan that experimental example 9 same operations obtain and join in the Eppendorf tube of capacity 1.5ml, it is mixed, obtain to contain the material of Pleurotus citrinopileatus extract and IOTA type carrageenan.
(8) experimental example 13
To contain with the Pleurotus citrinopileatus extract that the same operation of experimental example 8 obtains thing (solid component concentration 1%) 400 μ l and contain thing (solid component concentration 0.4%) 400 μ l with Testa oryzae/soybean fermented product that experimental example 10 same operations obtain and join in the Eppendorf tube of capacity 1.5ml, it is mixed, obtain to contain the material of Pleurotus citrinopileatus extract and Testa oryzae/soybean fermented product.
(9) experimental example 14
To contain with the IOTA type carrageenan that the same operation of experimental example 9 obtains thing 400 μ l and contain thing (solid component concentration 0.4%) 400 μ l with Testa oryzae/soybean fermented product that experimental example 10 same operations obtain and join in the Eppendorf tube of capacity 1.5ml, it is mixed, obtain to contain the material of I OTA type carrageenan and Testa oryzae/soybean fermented product.
(10) experimental example 15
To contain with the Pleurotus citrinopileatus extract that the same operation of experimental example 8 obtains thing (solid component concentration 1%) 400 μ l and contain thing (solid component concentration 2%) 400 μ l with Flos Rosae Multiflorae extract that experimental example 11 same operations obtain and join in the Eppendorf tube of capacity 1.5ml, it is mixed, obtain to contain the material of Pleurotus citrinopileatus extract and Flos Rosae Multiflorae extract.
(11) experimental example 16
To contain with the IOTA type carrageenan that the same operation of experimental example 9 obtains thing 400 μ l and contain thing (solid component concentration 2%) 400 μ l with Flos Rosae Multiflorae extract that experimental example 11 same operations obtain and join in the Eppendorf tube of capacity 1.5ml, it is mixed, obtain to contain the material of IOTA type carrageenan and Flos Rosae Multiflorae extract.
(12) experimental example 17
To contain with the Pleurotus citrinopileatus extract that the same operation of experimental example 8 obtains thing (solid component concentration 1%) 400 μ l and contain thing 400 μ l and contain thing (solid component concentration 0.4%) 400 μ l with Testa oryzae/soybean fermented product that experimental example 10 same operations obtain with IOTA type carrageenan that experimental example 9 same operations obtain and join in the Eppendorf tube of capacity 1.5ml, it is mixed, obtain to contain the material of Pleurotus citrinopileatus extract, IOTA type carrageenan and Testa oryzae/soybean fermented product.
(13) experimental example 18
To contain with the Pleurotus citrinopileatus extract that the same operation of experimental example 8 obtains thing (solid component concentration 1%) 400 μ l and contain thing 400 μ l and contain thing (solid component concentration 2%) 400 μ l with Flos Rosae Multiflorae extract that experimental example 11 same operations obtain with IOTA type carrageenan that experimental example 9 same operations obtain and join in the Eppendorf tube of capacity 1.5ml, it is mixed, obtain to contain the material of Pleurotus citrinopileatus extract, IOTA type carrageenan and Flos Rosae Multiflorae extract.
[table 2]
[4] piliation synergy evaluation
Above-mentioned experimental example 12～18 contains multiple extract (Multiple components), therefore adopts following method to inquire into the synergy that is brought by multiple extract.Being confirmed to be by adopting following method to calculate following value of calculation (the area value of calculation of the part that chaeta is few) of synergy, and compare with each experimental example and to carry out.
Value of calculation 1: the addition meansigma methods of experimental example 8 and 9 as value of calculation 1, is compared with the calculation and object value of experimental example 12.
Value of calculation 2: the addition meansigma methods of experimental example 8 and 10 as value of calculation 2, is compared with the calculation and object value of experimental example 13.
Value of calculation 3: the addition meansigma methods of experimental example 9 and 10 as value of calculation 3, is compared with the calculation and object value of experimental example 14.
Value of calculation 4: the addition meansigma methods of experimental example 8 and 11 as value of calculation 4, is compared with the calculation and object value of experimental example 15.
Value of calculation 5: the addition meansigma methods of experimental example 9 and 11 as value of calculation 5, is compared with the calculation and object value of experimental example 16.
Value of calculation 6: experimental example 8,9 and 10 addition meansigma methods as value of calculation 6, are compared with the calculation and object value of experimental example 17.
Value of calculation 7: experimental example 8,9 and 11 addition meansigma methods as value of calculation 7, are compared with the calculation and object value of experimental example 18.
In addition, the poor (Δ of the area value of above-mentioned value of calculation, above-mentioned value of calculation and experimental example 6 2Value), the Δ of object experiment and object experiment 1The Δ of value and value of calculation 2The difference of value is not shown in table 3.
[table 3]
Figure BDA0000060368290000261
In addition, by the Δ of above-mentioned value of calculation 2The Δ of value and the experiment of each object 1Ratio (the Δ of each embodiment of value 1The Δ of value/each value of calculation 2Value) estimates synergy.The Δ of above-mentioned value of calculation 2The Δ of value and the experiment of each object 1The ratio of value is shown in table 4.
[table 4]
The hair growth synergy
The Δ of embodiment 12 1The Δ of value/value of calculation 1 2Value 1.13
The Δ of embodiment 13 1The Δ of value/value of calculation 2 2Value 0.82
The Δ of embodiment 14 1The Δ of value/value of calculation 3 2Value 0.05
The Δ of embodiment 14 1The Δ of value/value of calculation 3 2Value 0.90
The Δ of embodiment 15 1The Δ of value/value of calculation 4 2Value 0.80
The Δ of embodiment 16 1The Δ of value/value of calculation 5 2Value 1.59
The Δ of embodiment 17 1The Δ of value/value of calculation 6 2Value 1.23
From the results shown in Table 1, the IOTA type carrageenan of the Flos Rosae Multiflorae extract of experimental example 1, the Testa oryzae/soybean fermented product of experimental example 2 and experimental example 3 is compared with the Pleurotus citrinopileatus extract of experimental example 4, and is high with the affinity of FGF-5.Can find out that particularly the KD value (M) of the IOTA type carrageenan of experimental example 3 is 2.95 * 10 -8, excellent with the affinity of FGF-5, good to the inhibition of FGF-5.
From the results shown in Table 2, compare with the experimental example 6 of 50% ethanol, experimental example 8～18 few without gross area, the hair growth effect of contained extract etc. is good in the experimental example 8～18.And then can find out, compare with experimental example 7 (minoxidil), experimental example 8 (Pleurotus citrinopileatus extract), experimental example 9 (IOTA type carrageenan), experimental example 11 (Flos Rosae Multiflorae extract) few without gross area, they have the hair growth effect more excellent than minoxidil.
In addition, from the results shown in Table 4, the experimental example 17 (Pleurotus citrinopileatus extract, IOTA type carrageenan and Testa oryzae/soybean fermented product), the experimental example 18 (Pleurotus citrinopileatus extract, IOTA type carrageenan and Flos Rosae Multiflorae extract) that contain 3 kinds of compositions, the value of their piliation synergy is high.This shows that the situation that respectively contains composition with independent use is compared, can bring into play more excellent hair growth effect by the synergy of 3 kinds of compositions.
To sum up, it is active high that the FGF-5 of Flos Rosae Multiflorae extract, Testa oryzae/soybean fermented product and IOTA type carrageenan suppresses, and the hair growth effect of Pleurotus citrinopileatus extract is good, therefore, the alopecia preventing effectiveness, piliation effect, foster hair (hair maintenance) effect and the long hair effect excellence that contain their hair growth agent.

Claims (5)

1.FGF-5 composite inhibiting is characterized in that, contains IOTA type carrageenan, Pleurotus citrinopileatus extract and Testa oryzae/soybean fermented product,
Described Testa oryzae/soybean fermented product obtains by making with extra care after using bacillus subtilis that Testa oryzae and soybean peptide are fermented again.
2. FGF-5 composite inhibiting claimed in claim 1, described bacillus subtilis is Bafillus natt.
3. claim 1 or 2 described FGF-5 composite inhibitings also contain the petal extract of Rosaceae gul.
4. hair growth agent is characterized in that, contains each described FGF-5 composite inhibiting of claim 1～3.
5. the livestock animals topical composition is characterized in that, contains each described FGF-5 composite inhibiting of claim 1～3.
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