CN110731922A

CN110731922A - Enhanced hair loss prevention and hair growth promoting composition

Info

Publication number: CN110731922A
Application number: CN201910693937.4A
Authority: CN
Inventors: 赵瑞丽; 王莎莎
Original assignee: Zhejiang Yangshengtang Institute of Natural Medication Co Ltd
Current assignee: Zhejiang Yangshengtang Institute of Natural Medication Co Ltd
Priority date: 2019-07-30
Filing date: 2019-07-30
Publication date: 2020-01-31
Anticipated expiration: 2039-07-30
Also published as: WO2021017911A1; CN110731922B

Abstract

The present invention provides enhanced hair loss prevention and growth promotion compositions comprising (a) concentrated birch sap and (B) a combination of any or more of ginger extract, polygonum multiflorum root extract, ginseng extract, green tea extract, arborvitae extract, angelica extract, astragalus, eclipta alba, birch sprout extract, birch leaf extract, adenosine, niacinamide, tripeptide-1 copper, wherein the concentration of the concentrated birch sap is about 1.2-8 times, preferably about 1.5-6 times, more preferably about 1.5-5 times.

Description

Enhanced hair loss prevention and hair growth promoting composition

Technical Field

The present invention relates to enhanced hair loss prevention and growth promotion compositions comprising (a) concentrated birch sap and (B) a combination of any or more of ginger extract, polygonum multiflorum root extract, ginseng extract, green tea extract, arborvitae extract, angelica extract, astragalus, eclipta alba, birch sprout extract, birch leaf extract, adenosine, nicotinamide, tripeptide-1 copper, wherein the concentration of the concentrated birch sap is about 1.2-8 times, preferably about 1.5-6 times, more preferably about 1.5-5 times.

Background

With the acceleration of the pace of life of modern society and the gradual worsening of the environment, more and more people have sub-health problems of different degrees under the conditions of high-intensity mental stress, overtime, overnight stay, haze and the like, wherein the problems include the increased secretion of scalp grease, scalp inflammation, hair follicle atrophy, hair loss and the like caused by the problems.

Disclosure of Invention

the present invention relates to the use of a combination of (a) concentrated birch juice and (B) ginger extract, fleece-flower root extract, ginseng extract, green tea extract, arborvitae extract, angelica extract, astragalus root, eclipta alba, birch sprout extract, birch leaf extract, adenosine, nicotinamide, tripeptide-1 copper, any or more, in an enhanced hair loss prevention and hair growth composition, wherein the concentrated birch juice is concentrated from about 1.2 to 8 times, preferably from about 1.5 to 6 times, more preferably from about 1.5 to 5 times.

In another aspect, the present invention provides enhanced hair loss prevention and growth promotion compositions comprising (a) concentrated birch sap and (B) any or more of ginger extract, polygonum multiflorum root extract, ginseng extract, green tea extract, arborvitae extract, angelica extract, astragalus root, eclipta alba, birch bud extract, birch leaf extract, adenosine, nicotinamide, tripeptide-1 copper, wherein the concentrated birch sap is concentrated about 1.2-8 times, preferably about 1.5-6 times, more preferably about 1.5-5 times.

The anti-alopecia hair growth relates to two key aspects, wherein is the activity of 5 α -reductase, inhibits the activity of 5 α -reductase, can effectively reduce the generation of dihydrotestosterone, and avoids the atrophy of hair follicles, and secondly, the activity of hair papilla cells is strong, the proliferation activity of the hair papilla cells can influence the state of the whole hair follicles, so that the hair follicles can keep the state of a growth period, promote the regeneration of hairs, and prolong the growth time of the hairs.

Unexpectedly, the present inventors found that, compared to the use of concentrated birch juice or ginger extract, polygonum multiflorum root extract, ginseng extract, green tea extract, arborvitae extract, angelica extract, astragalus root, eclipta alba, birch sprout extract, birch leaf extract, adenosine, nicotinamide, tripeptide-1 copper, the combination of concentrated birch juice with any or more of ginger extract, polygonum multiflorum root extract, ginseng extract, green tea extract, arborvitae extract, angelica extract, astragalus root, eclipta alba, birch sprout extract, birch leaf extract, adenosine, nicotinamide and tripeptide-1 copper has significantly better alopecia preventing and growing efficacy, much higher than the additive effect of the two functions, showing better inhibition of 5 α -reductase activity, regulation of transcription of hair papilla and alopecia preventing and growing-related genes and expression of proteins, promotion of proliferation and differentiation of hair papilla cells, and improvement of hair loss area of androgen model mice, which indicates that a synergistic effect occurs between the concentrated birch juice and the above substances.

Birch juices involved in the present invention are obtained from Betula genus of betulinaceae family, which may be from four varieties of white birch (Betula alba), Betula luminifera (Betula pubescens), Betula pendula (Betula pendula) and birch asiana (Betula platyphylla). The birch juice is colorless, transparent, precipitate-free and impurity-free, and has birch fragrance and rich nutrition, and is collected by manually drilling holes at the base of the birch trunk between thawing and early spring leaf emergence. The birch juice is commercially available and used as such, for example from greater Khingan over wild berry development, LLC.

The concentrated birch sap of the present invention is obtained by concentrating the above-mentioned commercially available products. Concentration methods are known in the art, such as heat concentration, low temperature vacuum concentration, membrane concentration, and the like. In the present invention, the concentration is preferably performed by a low-temperature freeze concentration or membrane concentration process, for example, commercially available birch juice stock solution is fed into a low-temperature drying device, cooled to-40 ℃ to-70 ℃, and vacuumized to 0.1 to 30Pa to perform low-temperature vacuum concentration, so as to obtain concentrated birch juice with different concentration times.

, the inventor also found that the anti-hair loss and hair growth promoting effect of the concentrated birch sap is not simply linear with its concentration degree, but shows a tendency of increasing and then decreasing with the increase of the concentration factor, therefore, it is critical to control the concentration factor of the birch sap, which is about 1.2-8 times, preferably about 1.5-6 times, and more preferably about 1.5-5 times in the present invention.

The content of the concentrated birch sap is about 10-98%, preferably about 20-98%, more preferably about 30-97%, based on the total weight of the enhanced hair loss prevention composition.

The component (B) ginger extract, polygonum multiflorum root extract, ginseng extract, green tea extract, thuja extract, angelica extract, astragalus root, eclipta alba, birch bud extract, birch leaf extract, adenosine, nicotinamide and tripeptide-1 copper are known in the art, and they are all commercially available as such and used in the present invention. The total content of the component (B) is about 0.0005 to 30%, preferably about 0.001 to 10%, more preferably about 0.1 to 5%, most preferably about 0.5 to 3%, based on the total weight of the hair loss prevention and hair growth promotion composition.

The hair loss prevention and hair growth promotion composition does not contain any added water, but does not exclude moisture inherently contained in the components.

Preferably, the hair loss preventing and hair growing composition does not contain a chelating agent such as EDTA salt, sodium polyphosphate, sodium metaphosphate, gluconic acid and the like.

The hair loss preventing and growing composition may optionally comprise, in addition to the above components (a) and (B), component (C), which is a component commonly used in hair care and shampoo compositions. Examples of the component (C) include, but are not limited to, vehicles, active ingredients, and adjuvants, and the like. These ingredients are known in the art and the type and amount of component (C) may be selected by those skilled in the art as desired, for example, the content of component (C) is generally about 0 to 70% based on the total weight of the hair loss prevention and hair growth promotion composition.

Such vehicles are known in the art and include, for example, diluents, dispersants or carriers and the like, examples of which include, but are not limited to, ethanol, dipropylene glycol, butylene glycol, and the like. The amount of vehicle in the composition is known in the art, and for example, it is generally 0.01 to 20% by weight of the total weight of component (C).

Such active ingredients are known in the art and include, for example, hair loss preventing hair restorers, moisturizers, hair conditioners and the like.

Examples of the hair loss prevention and hair growth agent include, but are not limited to, pubescent angelica root, angelica, ledebouriella root, fennel fruit, cnidium fruit, whiteflower hogfennel root, notopterygium root, perilla, scutellaria root, peppermint, wrinkled gianthyssop herb, fineleaf schizonepeta herb, lavender, sage, lime-pine, chrysanthemum, inula flower, cocklebur fruit, atractylodes rhizome, atractylodes macrocephala, snow lotus flower, elecampane, chamomile, groundsel, galangal, ginger, zedoary, turmeric, curcuma root, nutmeg, white peony root, moutan bark, cimicifuga foetida, ash bark, dittany bark, dried orange peel, citrus reticulata, finger citron, hawthorn leaf, hairyvein agrimony, asarum, madder, gardenia, sweet orange, mangostemon, veratrum, thyme, artemisia, valerian, prunella vulgaris, purslane, hirsutella, loranthus parasiticus, polygonum cuspidatum, ligustrum lucidum, edelweiss, grape seed, clove, magnolia bark, magnolia flower, aniseed, cinnamon, lindera root, piper, pepper, gentian, spikenard, myrrh, magnolia bark, magnolia flower, lindera root, magnolia flower.

Examples of such humectants include, but are not limited to, or more of glycerol, diglycerol, butylene glycol, propylene glycol, 1, 3-propanediol, dipropylene glycol, 1, 2-pentanediol, polyethylene glycol-8, polyethylene glycol-32, methyl gluceth-10, methyl gluceth-20, PEG/PPG-17/6 copolymer, glyceryl polyether-7, glyceryl polyether-26, glyceryl glucoside, PPG-10 methyl glucose ether, PPG-20 methyl glucose ether, PEG/PPG/polytetramethylene glycol-8/5/3 glycerol, sucrose, trehalose, rhamnose, mannose, raffinose, betaine, erythritol, xylitol, urea, glyceryl polyether-5 lactate, sodium hyaluronate, hydrolyzed sodium hyaluronate, acetylated sodium hyaluronate, sodium polyglutamate, hydrolyzed sclerotium rolfsii gum, pullulan, tremella polysaccharide, tamarind seed polysaccharide, and the like.

Examples of such hair conditioning agents include, but are not limited to, or more of fatty alcohols, sterols, fatty acids, monoglycerides, triglycerides, lanolin, sarcosinate, fatty acid esters, white oil, squalane, polyquaternium-10, polyquaternium-7, polyquaternium-39, polyquaternium-52, polyquaternium-73, guar hydroxypropyltrimonium chloride, behenyltrimethylammonium chloride, dimethicone, PEG-60 almond oil glyceride, panthenol, polyvinylpyrrolidone, gelatin, pectin, gleditsia sinensis extract, Salvia miltiorrhiza extract, Angelica sinensis root extract, butylene glycol, Polygonum multiflorum extract, Chamaecyparis leaf extract, ginger root extract, Scutellaria baicalensis root extract, Picrasma kurarianum root extract, Glycyrrhiza inflata root extract, allantoin, Panax notoginseng root extract, Cochinchinensis extract, oat polypeptide, etc.

Such adjuvants are known in the art and include, for example, foaming agents, surfactants, emulsifiers, thickeners, preservatives, fragrances and the like.

Examples of such foaming agents include, but are not limited to, of sodium laureth sulfate, sodium trideceth sulfate, decyl glucoside, sodium lauroyl sarcosinate, sodium C14-16 olefin sulfonate, coconut monoethanolamide, coconut diethanolamide, cocoyl propyl betaine, ammonium lauryl sulfate, sodium cocoamphoacetate, and the like, the content of such foaming agents in the composition is known in the art, and for example, it is generally 0.1 to 50% by weight based on the total weight of component (C).

Examples of such surfactants include, but are not limited to, cocamidopropyl betaine, sodium laureth sulfate, PEG-150 distearate, ethylene glycol distearate, ammonium laureth sulfate, sodium laureth sulfate, palmamidopropyl betaine, cocamidodiethanol amine, lauryl ether sulfate, ammonium lauryl sulfate (K12A), sodium fatty alcohol polyoxyethylene ether sulfate (AES), ammonium fatty alcohol polyoxyethylene ether sulfate (AESA), ammonium lauryl ether sulfate, ammonium lauryl alcohol sulfate, sodium lauryl ether sulfate, ammonium laureth sulfate, ammonium lauryl alcohol sulfate, ammonium cocoyl alcohol sulfate, cocoyl monoethanolamide, N-fatty acyl amino acid salts, lauramidopropyl betaine, sodium cocoyl amphoacetate, sodium lauroamphoacetate, sodium laureth sulfate, sodium fatty alcohol polyoxyethylene ether sulfate, sodium lauryl sulfate, alkyl glycosides, dodecyl dimethyl betaine, disodium imidazolinium diacetate, sodium cocoamphoacetate, polyquaternary ammonium salt-7, polyquaternary ammonium salt-10, polyquaternary ammonium salt-37, hydroxypropyl guar trimethyl chloride, hydroxypropyl trimethyl ammonium chloride, guar, guanidium extract, etc. or more of such surfactants are typically present in the compositions at a total weight of such as 0.1 to 50% of the surfactant.

Examples of such emulsifiers include, but are not limited to, cetearyl olivate, sorbitan olivate, polysorbate-60, polysorbate-80, methylgluco-sesquistearate, PEG-20 methylgluco-sesquistearate, PEG-40 hydrogenated castor oil, PPG-26-butanoeth-26, PEG-4 polyglyceryl-2 stearate, PEG-60 hydrogenated castor oil, steareth-2, steareth-21, PPG-13-decyltetraeth-24, cetearyl glucoside, PEG-100 stearate, glyceryl stearate SE, coco glucoside, ceteareth-25, PEG-40 stearate, polyglyceryl-3 methylgluco distearate, glyceryl stearate citrate, polyglyceryl-10 stearate, polyglyceryl-10 myristate, polyglyceryl-10 dioleate, polyglyceryl-10 laurate, polyglyceryl-10 isostearate, polyglyceryl-10 oleate, polyglyceryl-10 diisostearate, polyglyceryl-6 laurate, polyglyceryl-6 myristate, sucrose % of the total weight of such emulsifiers, e.05% of the emulsifier is typically present in the composition.

Examples of such thickeners include, but are not limited to, of high molecular weight polymers such as carbomers, acrylates and derivatives thereof, xanthan gum, acacia, polyethylene glycol-14M, polyethylene glycol-90M, succinoglycan, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, and the like, the amount of such thickeners in such compositions is known in the art, and typically comprises from 0.1 to 30% by weight of component (C), for example.

Examples of such preservatives include, but are not limited to or more of methylparaben, ethylparaben, propylparaben, phenoxyethanol, benzyl alcohol, phenethyl alcohol, bis (hydroxymethyl) imidazolidinyl urea, potassium sorbate, sodium benzoate, chlorphenesin, sodium dehydroacetate, and the like, the amount of such preservatives in the hair loss prevention and growth promotion composition is known in the art, and for example, is typically 0.01 to 30% by weight of the total weight of component (C).

The anti-alopecia hair growth composition of the present invention can be prepared by any suitable method known in the art. For example, it is prepared using a dissolving tank, an emulsifying pot, a disperser, a transfer pump, etc., which are generally used in the art. The preparation method comprises putting water soluble substance into water phase dissolving kettle, putting oil soluble substance into oil phase dissolving kettle, heating the two kettles to about 80 deg.C, wherein the raw material easy to agglomerate can be pre-dispersed with disperser. After the dissolution is finished, the oil phase and the water phase are conveyed into an emulsifying pot, and homogenized and emulsified for about 5-15 minutes. After emulsification is finished, the temperature of the material body is reduced to normal temperature, optional essence, preservative and the like are added, and the pH of the product is adjusted according to needs. After the relevant detection indexes are qualified, the products can be filled and delivered. The preparation method can be deleted or adjusted according to the requirements of dosage forms.

The hair loss preventing and growing composition of the present invention may be in the form of a shampoo for washing off, or a hair tonic for leave-on, and it may be prepared in various forms such as cream, milky lotion, liquid, etc., as required.

Examples

The invention is described in further detail at with reference to the following examples, however, it should be understood that these examples and comparative examples are intended only to illustrate the invention in more detail and should not be construed as limiting in any way the scope of the appended claims.

Example 15 α inhibition of reductase Activity

In this example, the effect of concentrated birch juice, ginger extract, polygonum multiflorum root extract, ginseng extract, green tea extract, thuja extract, and combinations thereof on the activity of 5 α -reductase was examined and compared.

1. Concentration of birch sap

Fresh birch sap stock solution purchased from Daxingan mountain surpassing the company Limited for wild berry development is introduced into a low-temperature drying device, cooled to-65 ℃, vacuumized to 0.1Pa, and concentrated by 1.5 and 10 times.

2. Testing

An experimental instrument: balance, wall breaking machine, constant temperature oscillator, high speed freezing centrifuge, enzyme mark appearance.

Experimental reagents and consumables:

finasteride: shanghai modern pharmaceuticals, Inc. (tablets 5 mg/tablet);

reduced coenzyme ii (NADPH): roche, USA (powder 10 mg);

testosterone Elisa kit: wuhan Youerson company (Cat: CEA458 Ge);

5 α -reductase crude enzyme, self-prepared;

PBS: wuhan doctor de Corp;

phenylmethylsulfonyl fluoride (PMSF): sigma, USA;

dithiothreitol (DTT): sigma, USA;

1M Tris-HCl: wuhan doctor de Corp.

Sample loading information:

the loading amount of the raw material of the single is parts of the whole raw material, and the compound raw material is half of the raw material of the birch juice and half of 0.1 percent of other active raw materials.

In vitro 5 α -reductase activity impact assay procedure was as follows:

(1) preparing a crude enzyme: taking 5 normal healthy mice, dislocating and killing the neck, opening the abdominal cavity, taking testis, dividing into several parts, placing in a 2mL EP tube, adding a proper amount of crude enzyme extract according to a ratio of 1:4, crushing by a wall breaking machine at 4 ℃ to prepare homogenate, freezing and centrifuging at 4 ℃ at a high speed (10000 rpm/min, 10 min), and taking supernatant and storing at 4 ℃. The BCA method is used for determining the protein concentration, and the subsequent experiment can be carried out when the protein concentration is more than 1 mg/mL;

(2) measuring blank control, namely respectively taking 2 groups of 200 mu L EP tubes (4 per group), adding 5 α -reductase crude enzyme, PBS solution (pH is 7.4) and reduced coenzyme II, mixing groups, immediately putting into boiling water for boiling for 5 minutes, centrifuging, absorbing 50 mu L of liquid for subsequent Elisa detection to obtain initial testosterone content of the blank control group, mixing groups, putting into a constant-temperature shaking table for uniformly mixing for 60 minutes, putting into boiling water for boiling for 5 minutes, centrifuging, absorbing 50 mu L of liquid for subsequent Elisa detection, wherein the difference value between the initial content and the initial content is the converted testosterone content of the blank control group;

(3) the determination of the sample group is that 2 groups of 200 mu L EP tubes (4 per group) are respectively taken, 5 α -reductase crude enzyme, PBS solution (pH 7.4), reduced coenzyme II and test raw materials are added, groups are mixed and immediately put into boiling water to be boiled for 5 minutes, 50 mu L of liquid is absorbed after centrifugation to carry out subsequent Elisa detection, the initial testosterone content of the inhibitor group is obtained, another groups are mixed and then put into a constant temperature shaking table to be mixed for 60 minutes, and put into boiling water to be boiled for 5 minutes, 50 mu L of liquid is absorbed after centrifugation to carry out subsequent Elisa detection, the difference value between the initial content and the initial content is the testosterone content after the transformation of the inhibitor group, and 4 parallel samples are obtained in each experiment.

The 5 α -reductase inhibition rate was calculated as follows:

I％＝(△A₀-△A_n)/△A₀*100％

wherein:

△A₀control testosterone reduction;

△A_ntestosterone reduction in the inhibitor group.

The test results are shown in the following table.

The results show that the effect of the combination of the 1.5 times of concentrated birch juice and the ginger extract, the tuber fleeceflower root extract, the ginseng extract, the green tea extract and the platycladus orientalis extract is obviously better than the effect of the combination of the birch juice stock solution, the 1.5 times of concentrated birch juice and the ginger extract, the tuber fleeceflower root extract, the ginseng extract, the green tea extract and the platycladus orientalis extract which are singly used in the aspect of inhibiting 5 α -reductase, and simultaneously the effect of the combination of the birch juice stock solution and the ginger extract, the tuber fleeceflower root extract, the ginseng extract, the green tea extract and the platycladus orientalis extract is also obviously better than the effect of the combination of the birch juice stock solution and other active matter raw materials.

Example 2: influence on the expression of hair loss prevention and hair growth related genes and proteins

In this example, the effects of concentrated birch juice, angelica extract, astragalus, eclipta alba, birch bud extract, birch leaf extract, and combinations thereof on the expression of hair loss prevention and hair growth related genes and proteins were examined and compared.

1. Concentration of birch sap

2. Testing

The dermal papilla cells used in this experiment were produced by Dongbo xi Biotech limited.

An experimental instrument: CO 2₂Incubator (Thermo), clean bench (Sujing Antai), inverted microscope (Olympus), micro-oscillator (Linbel), enzyme labeling instrument (BioTek), incubator (Tester), fluorescent quantitative PCR instrument (BioRad), general PCR instrument (Bori).

Experimental reagent: mesenchymal Stem Cell Medium (ScienCell), PBS (Philid), DHT (Sigma), VEGF ELISA kit (Abcam), RNAiso Plus (Takara), reverse transcription kit (Takara), fluorescent dye (Takara).

The sample loading information comprises that the loading amount of the single raw material is parts of the whole raw material, and the compound raw material is half of the birch juice raw material and half of 0.1 percent of other active raw materials.

The steps of analysis of gene and protein expression based on hair papilla cells were as follows:

(1) inoculation: inoculating into 6-well plate at 37 deg.C and 5% CO₂Incubating in an incubator overnight;

(2) preparing liquid: the test article working solution was prepared according to the following experimental design table.

The experimental design is as follows:

(3) administration: according to the specific design of the experiment in the table above, when the plating rate of the cells in the 6-well plate reaches 40-50%, the administration amount of each well is 2mL, and each group is provided with 3 multiple wells. 37 ℃ and 5% CO₂The incubator continues to incubate for 24 hours.

(4) Cell supernatants and cells were collected separately:

a. collecting cell supernatant: after culturing for 24 hours, collecting cell culture supernatant in an EP tube, and after collection, placing a sample for VEGF content detection in a refrigerator at-80 ℃ for freezing and storing;

b. collecting cells: after 24 hours of culture, cell supernatants were collected, washed twice with 1 mL/well PBS, 1mL of NAiso Plus was added to each well, lysed cells were aspirated, and samples were collected.

(5) And (3) detection of VEGF content: the detection assay was performed according to the instructions of the VEGF ELISA kit.

(6) Detection of AR gene expression: extracting RNA, reverse transcribing to cDNA, and fluorescent quantitative PCR detection with 2^-△△CTThe method performs a result calculation.

3. Statistical analysis of results

The plots were generated using GraphPad Prism Program software and statistically analyzed between groups using T-test.

The test results are shown in the following table.

Sample (I)	VEGF protein (pg/mL)	AR Gene (relative transcription amount)
			Negative control	706±6	1.00±0.05
Stock solution of birch juice	814±6	0.82±0.03
			Concentrated birch juice 1.5 times	839±12	0.64±0.04
Concentrated birch juice 10 times	828±8	0.74±0.05
			Angelica sinensis extract	765±17	0.88±0.07
Radix astragali	728±23	0.91±0.06
			Herba Ecliptae	789±19	0.86±0.05
Birch bud	744±25	0.83±0.06
			White birch leaf	775±18	0.88±0.04
Birch juice stock solution and radix Angelicae sinensis extract	933±14	0.78±0.03
			Birch juice stock solution and radix astragali	956±22	0.75±0.06
Birch juice stock solution and eclipta alba	978±16	0.74±0.05
			Stock solution of birch juice and birch bud	991±21	0.76±0.07
Birch juice stock solution and birch leaves	948±18	0.75±0.01
			Concentrated birch juice 1.5 times and radix Angelicae sinensis extract	1265±45	0.38±0.04
1.5 times of concentrated birch juice and radix astragali	1309±31	0.37±0.03
			1.5 times of concentrated birch juice and eclipta alba	1257±28	0.41±0.05
1.5 times of concentrated birch juice and birch bud	1319±34	0.35±0.02
			1.5 times of concentrated birch juice and birch leaf	1301±27	0.43±0.04
Concentrated birch juice 10 times and radix Angelicae sinensis extract	887±36	0.79±0.03
			10 times of concentrated birch juice and astragalus root	843±28	0.78±0.06
10 times of concentrated birch juice and eclipta alba	865±27	0.74±0.05
			Concentrated birch juice 10 times and birch bud	896±41	0.80±0.04
Concentrated birch juice 10 times and birch leaf	870±38	0.75±0.07

The above results show that compared with the use of the birch juice stock solution, the 1.5-fold concentrated birch juice, the angelica sinensis extract, the astragalus membranaceus, the eclipta alba, the birch bud and the birch leaves alone, and the combination of the birch juice stock solution and the angelica sinensis extract, the astragalus membranaceus, the eclipta alba, the birch bud or the birch leaves, the use of the 1.5-fold concentrated birch juice in combination with the angelica sinensis extract, the astragalus membranaceus, the eclipta alba, the birch bud or the birch leaves significantly increases the expression level of VEGF protein (vascular endothelial growth factor) related to the microcirculation of blood vessels around papillae, and effectively inhibits the transcription of AR (androgen receptor) genes related to androgen signal transmission. When the birch juice is concentrated to 10 times, the effect of combining the birch juice with other active matters is obviously weakened or even not as good as the effect of combining the stock solution birch juice with other active matter raw materials.

Example 3: effect on the proliferation and differentiation Capacity of Hair papilla cells

In this example, the effect of various fold-concentrated birch sap, adenosine, nicotinamide, tripeptide-1 copper, and their combinations on the proliferation and differentiation capacity of hair papilla cells was examined and compared.

1. Concentration of birch sap

2. Test method

An experimental instrument: CO 2₂Incubator (Thermo), clean bench (Sujing Antai), inverted microscope (Olympus), micro-oscillator (Linbel), enzyme labeling instrument (BioTek), and incubator (Tester).

Experimental reagents and consumables: mesenchymal Stem Cell Medium (science Cell), PBS (Phd.), MTT (Sigma), DMSO (Sigma).

The experimental method comprises the following steps:

(1) preparing a cell suspension: digesting cells in logarithmic growth phase, inoculating the cells into a 96-well plate, and inoculating the cells into the 96-well plate at 37 ℃ and 5% CO₂Incubating in an incubator overnight;

(2) administration: when the cell plating rate in the 96-well plate reaches 20-30%, the prepared culture solution with the sample to be detected is administered in groups, the administration amount of each hole is 200 mu L, and each group is provided with 3 multiple holes. MTT assay was performed on 0 hour-grouped cells simultaneously, and at other time points, the cells were incubated at 37 ℃ in 5% CO₂Continuously culturing in an incubator;

(3) and (3) detection: the cells were incubated for 48 hours, the supernatant was discarded, MTT working solution was added, and incubation was carried out at 37 ℃ in the dark. Discarding the supernatant after 4 hours, adding 150 mu L DMSO into each hole, and reading the OD value by an enzyme-labeling instrument at 490 nm;

(4) calculating the formula:

the test results are shown in the following table.

Sample (I)	Rate of hair papilla cell proliferation
		Stock solution of birch juice	18％±2％
Concentrated birch juice 1.5 times	33％±7％
		Concentrated birch juice 10 times	22％±3％
Adenosine (I)	6％±3％
		Nicotinamide	13％±5％
Tripeptide-1 copper	8％±3％
		Birch juice stock solution + adenosine	23％±4％
Birch juice stock solution and nicotinamide	24％±6％
		Birch juice stock solution + tripeptide-1 copper	26％±5％
1.5 times of concentrated birch juice + adenosine	52％±4％
		1.5 times of concentrated birch juice + nicotinamide	48％±7％
1.5 times of concentrated birch juice and tripeptide-1 copper	49％±4％
		10 times concentrated birch juice + adenosine	22％±4％
10 times of concentrated birch juice and nicotinamide	22％±3％
		10 times concentrated birch juice + tripeptide-1 copper	23％±3％

MTT value test results show that compared with the single use of birch juice stock solution, 1.5 times of concentrated birch juice, adenosine, nicotinamide and tripeptide-1 copper and the combination of the birch juice stock solution and the adenosine, the nicotinamide and the tripeptide-1 copper, the combination of the 1.5 times of concentrated birch juice and the tripeptide-1 copper obviously improves the proliferation and differentiation capacity of hair papilla cells. When the birch juice is concentrated to 10 times, the effect of combining the birch juice with other active matters is obviously weakened or even not as good as the effect of combining the stock solution birch juice with other active matter raw materials.

Example 4: comparison of Effect on mouse models

1. Concentration of birch sap

2. Experimental methods

Experimental animals: c57BL/6 mice, 6-7 weeks old, male; the animal is from Beijing vitamin Tonglihua.

The determination method was that about 15C 57BL/6 mice per group were assigned to the environment for 1 week and then divided into groups according to the weight. Removing hair from back with depilator, and uniformly applying depilatory cream to back at a speed of about 3x4cm²And (3) washing after a plurality of minutes, inducing the hair follicles of the mice to grow into a growth phase, wherein the skin of the mice is pink, injecting testosterone propionate with the dose of 5 mg/kg/day subcutaneously at two sides () of the animals of other groups except a normal control group 1 time every day, injecting soybean oil with the same volume subcutaneously in the normal control group simultaneously by molding, and measuring the hair removal area of the backs of the mice after smearing 0.1mL for 2 months.

The test results are shown in the following table.

Sample (I)	Depilatory area (Pixel)
		Blank space	10183±92
Stock solution of birch juice	9105±47
		Concentrated birch juice 1.5 times	7466±90
Concentrated birch juice 10 times	9005±85
		Polygonum multiflorum root extract	9819±43
Green tea extract	9836±64
		Birch juice stock solution and radix Polygoni Multiflori extract	7967±59
Birch juice stock solution and green tea extract	8165±39
		Concentrated birch juice 1.5 times and radix Polygoni Multiflori extract	5173±93
Concentrated birch juice 1.5 times and green tea extract	4032±75
		Concentrated birch juice 10 times and radix Polygoni Multiflori extract	8467±59
Concentrated birch juice 10 times and green tea extract	8565±39

Mouse model experimental results show that compared with the single use of the birch juice stock solution, the 1.5 times of concentrated birch juice, the polygonum multiflorum root extract and the green tea extract and the combination of the birch juice stock solution and the polygonum multiflorum root extract or the green tea extract, the combined use of the 1.5 times of concentrated birch juice and the polygonum multiflorum root extract or the green tea extract obviously reduces the unhairing area of a mouse and promotes the rapid growth of new hair. When the birch juice is concentrated to 10 times, the effect of combining the birch juice with other active matters is obviously weakened or even not as good as the effect of combining the stock solution birch juice with other active matter raw materials.

Example 5: hair loss preventing and hair growing liquid

The formula of the anti-hair loss and hair growth liquid is shown in the following table:

the preparation method of the hair restorer comprises the following steps:

1. firstly, adding the phase A components into a main pot according to the mass ratio, heating to 80 ℃, and mixing;

2. adding the components of the phase C into a side pot according to the mass ratio, heating to melt and uniformly stirring, putting the mixture into the phase A, and uniformly stirring and mixing the mixture with the phase A;

3. sequentially adding the amino acid, the nutritional supplement, the oil control agent and the nicotinamide in the C phase into the main pot, and uniformly stirring;

4. sequentially adding the diaminopyrimidine oxide in the phase D and the solvent in the phase D into the main pot, and uniformly stirring;

5. cooling the main kettle to 50 ℃, sequentially adding the alcohol in the phase E and the solvent in the phase E, and uniformly stirring;

6. cooling the main kettle to 45 ℃, adding the tripeptide-1 copper of the F phase, and uniformly stirring;

7. mixing Tween-20 and essence in the G phase, pre-dissolving, stirring to transparent, adding into the main pot, and stirring;

8. and finally, adding the H phase into the main pot, uniformly stirring, cooling to room temperature, and discharging after the detection is qualified to obtain the hair growth liquid for preventing alopecia.

The clinical human body test comprises selecting 120 hair loss patients (the hair loss number is more than 100 per day), randomly dividing into six groups, respectively using the six shampoos, wherein each group is half of male and female, applying the hair on the scalp every day, massaging and absorbing, combing the hair 2 times in the morning and evening, collecting and counting the hair loss, recording times per week, continuously using and recording for 4 weeks, and obtaining the following results:

the above results of the anti-sloughing solution show that the combination of the concentrated birch juice of 1.5 times and the tripeptide-1 copper has more remarkable anti-sloughing effect compared with the single use of the birch juice stock solution, the concentrated birch juice of 1.5 times, the tripeptide-1 copper and the combination of the birch juice stock solution and the tripeptide-1 copper.

Example 6: hair loss preventing and hair growing shampoo

The formula of the hair loss preventing and hair growing shampoo is shown in the following table:

the preparation method of the shampoo comprises the following steps:

deionized water or birch sap was added in the proportions indicated in the table and heated to 80 c for half an hour. Sequentially adding sodium lauryl alcohol ether sulfate, sodium cocoyl glutamate, guar gum and birch sprout into water, continuously stirring, and keeping for 10 minutes. Homogenizing the mixture for 20-30 minutes; mixing 1, 2-pentanediol, 1,3-CG, D-panthenol, menthol and eucalyptus leaf oil in advance, adding the mixture into the system when the temperature of the homogeneous mixture is reduced to room temperature, and stirring for 10 minutes. Citric acid and sodium chloride were added. The pH was adjusted to 5-7 and finally deionized water was added to 100% of the recipe weight. Filtering with 1200 mesh, and bottling to obtain shampoo.

The clinical human body test comprises the steps of selecting 120 alopecia patients (the number of alopecia is more than 100 per day), randomly dividing the alopecia patients into six groups, using the six shampoos respectively, wherein each group is half of a male and a female, a subject washes times every days, combs hair 2 times in the morning and evening, collects and counts the fallen hair, records times every 15 days, and continuously uses and records for 3 months to obtain the following results:

the use results show that compared with the single use of the birch sap stock solution, the 1.5-time concentrated birch sap, the birch bud and the combination of the birch sap stock solution and the birch bud, the combination of the 1.5-time concentrated birch sap and the birch bud has more remarkable anti-dropping effect.

The technical solutions of the above-described embodiments are preferred embodiments of the present invention, and several modifications and changes can be made without departing from the principle of the present invention, and these modifications and changes should also be considered as being within the protection scope of the present invention.

Claims

Use of (a) concentrated birch juice in combination with (B) any or more of ginger extract, polygonum multiflorum root extract, ginseng extract, green tea extract, arborvitae extract, angelica extract, astragalus root, eclipta alba, birch bud extract, birch leaf extract, adenosine, nicotinamide and tripeptide-1 copper in an anti-alopecia hair growth composition, wherein the concentration factor of the concentrated birch juice is 1.2-8, preferably 1.5-6, more preferably 1.5-5.
2. The use of claim 1, wherein said alopecia preventing and growing composition does not comprise any added water.
Hair loss prevention and growth promoting composition comprising (A) concentrated birch sap and (B) ginger extract, Polygonum multiflorum root extract, Panax ginseng extract, Green tea extract, Platycladus orientalis extract, Angelica sinensis extract, Astragalus membranaceus, Ecliptae herba, birch sprout extract, birch leaf extract, adenosine, nicotinamide and tripeptide-1 copper at any or more, wherein the concentration of the concentrated birch sap is 1.2-8, preferably 1.5-6, more preferably 1.5-5.
4. The composition of claim 3, wherein said alopecia preventing composition does not comprise any added water.
5. The composition according to claim 3 or 4, wherein the content of the concentrated birch sap of component (A) is 10-98%, preferably 20-98%, more preferably 30-97%, based on the total weight of the alopecia preventing and growing composition.
6. The composition of any one of claims 3 to 5, , wherein the component (B) is present in an amount of 0.0005 to 30%, preferably 0.001 to 10%, more preferably 0.1 to 5%, most preferably 0.5 to 3%, based on the total weight of the anti-alopecia composition.