CN113908091B - Anti-hair-loss hair-growing composition and preparation method and application thereof - Google Patents
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- CN113908091B CN113908091B CN202111198925.8A CN202111198925A CN113908091B CN 113908091 B CN113908091 B CN 113908091B CN 202111198925 A CN202111198925 A CN 202111198925A CN 113908091 B CN113908091 B CN 113908091B
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Classifications
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/60—Sugars; Derivatives thereof
- A61K8/602—Glycosides, e.g. rutin
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/63—Steroids; Derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/14—Drugs for dermatological disorders for baldness or alopecia
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- A—HUMAN NECESSITIES
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
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- A61Q5/008—Preparations for oily hair
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q5/00—Preparations for care of the hair
- A61Q5/02—Preparations for cleaning the hair
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q5/00—Preparations for care of the hair
- A61Q5/12—Preparations containing hair conditioners
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
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Abstract
The invention provides an anti-hair loss hair-growing composition, and a preparation method and application thereof. The hair-loss preventing hair-growing composition mainly comprises the following components: glycyrrhizin, boerduo leaf extract, oleanolic acid and stilbene glucoside. The four components cooperate and synergistically, so that the activity of 5 alpha-reductase is effectively inhibited, the conversion of testosterone into DHT by 5 alpha-reductase is avoided to a great extent, and the problems of rapid entry into the retrograde phase, micromation, hair loss and the like caused by the action of high-activity DHT on hair follicles after the combination of the high-activity DHT and an androgen receptor are avoided; meanwhile, the hair loss prevention and hair growth promotion composition can also inhibit the expression of IL-1 beta and TNF-alpha inflammatory factors in skin hair follicles, so that the purposes of preventing hair loss and promoting hair growth are finally achieved.
Description
Technical Field
The invention belongs to the technical field of cosmetics, and particularly relates to an anti-hair-loss hair-growing composition, and a preparation method and application thereof.
Background
Hair is an accessory of human body, is an important component of human body, and has protection, modification and other physiological functions. And alopecia is a skin disease characterized by hair loss. Hair loss causes sparse hair or premature baldness, which can affect the appearance, brings much trouble and pain to patients, affects the normal work and life of people, and even destroys the mental health of patients, hits the self-confidence of people, and healthy maintenance of hair to prevent hair loss is an urgent problem for people to solve.
In the case of alopecia, however, one of the biggest causes is abnormal secretion of sebaceous glands. For abnormal sebaceous gland secretion, medicine is generally aimed at androgen excess, and estrogen regulation is increased or androgen secretion is reduced by using some drugs for abnormal sebaceous gland secretion caused by androgens, such as cimetidine, and 5 alpha-reductase is inhibited by using finasteride, dutasteride and irinotecan. However, the use of regulatory hormonal drugs has different injuries to the body. Therefore, the safety of the 5α -reductase inhibitory drugs of the prior art has yet to be improved. And for nursing oily skin, the skin is mainly cleaned thoroughly, the secretion of grease is controlled, and the skin is in a water-oil balance state as much as possible. Meanwhile, the anti-inflammatory and antibacterial effects are achieved, and skin diseases such as acne and seborrheic dermatitis can be effectively prevented and controlled.
CN110025775a discloses a hair-loss preventing and hair-growing essence for warming and curing hair and a preparation method thereof, wherein the hair-loss preventing and hair-growing essence for warming and curing hair comprises the following raw materials: water, ethanol, stilbene glucoside, glycerol, angelica sinensis extract, aloe extract, ginseng extract, tea polyphenol, corn gluten amino acids, vitamin H, folic acid, vitamin B3, pantothenic acid, vitamin B6, vitamin B2, vitamin B1, yeast polypeptides, cysteine, sodium benzoate, potassium sorbate, citric acid, linoleic acid, xylanase and hydroid extract. According to the invention, the linoleic acid, xylanase and hydroid extracts are adopted to synergistically improve the alopecia preventing rate and the hair growth promoting rate of the hair loss preventing and hair growth promoting essence for mildly fixing the hair.
CN103446563a discloses a plant extract compound for preventing alopecia, which inhibits 5 alpha-reductase activity, and belongs to plant extract compositions. The hair loss preventing compound comprises saw blade palm extract, waxberry skin extract, biota orientalis extract, angelica sinensis extract, rhizoma anemarrhenae extract, selfheal extract, ginger extract, radix sophorae flavescentis extract, jackfruit extract, schisandra chinensis extract and kaempferia galanga extract, wherein the weight percentage of each plant extract in the total amount of the hair loss preventing compound is 0.1-99%. The anti-hair loss plant compound is used for treating and improving seborrheic alopecia symptoms caused by male androgenetic sex.
Therefore, developing a head care product that fundamentally solves the problem of hair loss is an important research focus in the art.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide an anti-hair loss hair-growing composition, and a preparation method and application thereof. The anti-hair loss hair-growing composition achieves the effects of controlling oil, inhibiting inflammatory factors and resisting inflammation and inhibiting bacteria by inhibiting the activity of 5 alpha-reductase, can realize the water-oil balance of scalp and effectively restore the barrier function of scalp, thereby radically solving the problem of hair loss.
To achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the present invention provides an anti-hair-growing composition comprising: glycyrrhizin, boerduo leaf extract, oleanolic acid and stilbene glucoside.
According to the invention, through the mutual coordination of the glycyrrhizin, the Boldol leaf extract, the oleanolic acid and the stilbene glucoside, the synergistic effect is achieved, so that the activity of 5 alpha-reductase is effectively inhibited, the conversion of testosterone into DHT by 5 alpha-reductase is avoided to a great extent, and the problems of rapid entry into the retrograde phase, micromation, hair loss and the like caused by acting on hair follicles after the combination of high-activity DHT and androgen receptor are avoided; meanwhile, the hair loss prevention and hair growth promotion composition can also inhibit the expression of IL-1 beta and TNF-alpha inflammatory factors in skin hair follicles, so that the purposes of preventing hair loss and promoting hair growth are finally achieved.
In the invention, the glycyrrhizin can inhibit the occurrence of peptic ulcer, prevent premature graying and alopecia, and can obviously improve the anti-hair loss effect when being added into the hair care anti-hair loss and hair growth composition, and has the effects of diminishing inflammation, inhibiting bacteria, resolving masses and relieving exterior syndrome.
In the invention, the Boldol leaf extract activates the synthesis of skin beta-defensin (an endogenous antibacterial peptide), enhances the natural antibacterial system of skin, reduces bacterial growth, inhibits inflammatory factors, fine pores and improves oily scalp.
In the invention, oleanolic acid can inhibit 5 alpha-reductase activity so as to achieve the oil control effect, and has the advantages of quick oil control effect, long oil control time and difficult repetition.
In the invention, the main component of the stilbene glucoside comprises the stilbene glucoside, the name of the stilbene glucoside is 2,3,5,4' -tetrahydroxy stilbene-2-O-beta-D-glucoside, the stilbene glucoside belongs to polyhydroxy stilbene compounds, and the stilbene glucoside can permeate into hair roots and has the effects of sterilization, bacteriostasis and antioxidation.
Preferably, the anti-hair loss hair-growing composition comprises the following components in parts by weight: 0.01-1 part of liquiritin, 1-3 parts of Boldol tree leaf extract, 0.01-2 parts of oleanolic acid and 0.1-0.5 part of stilbene glucoside.
The content of the liquiritin in the hair-growing composition for preventing alopecia is 0.01-1 part, for example, 0.01 part, 0.02 part, 0.05 part, 0.08 part, 0.1 part, 0.2 part, 0.3 part, 0.4 part, 0.5 part, 0.6 part, 0.7 part, 0.8 part, 0.9 part, 1 part and the like.
In the hair-loss preventing and hair-growing composition, the content of the Boerdos leaf extract is 1-3 parts, for example, 1 part, 1.2 parts, 1.4 parts, 1.6 parts, 1.8 parts, 2 parts, 2.2 parts, 2.4 parts, 2.6 parts, 2.8 parts, 3 parts and the like can be used.
The content of oleanolic acid in the hair-growing composition is 0.01-2 parts, for example, 0.01 parts, 0.05 parts, 0.1 parts, 0.2 parts, 0.3 parts, 0.4 parts, 0.5 parts, 0.6 parts, 0.8 parts, 1 parts, 1.2 parts, 1.4 parts, 1.6 parts, 1.8 parts, 2 parts and the like.
The content of stilbene glucoside in the hair-growing composition is 0.1-0.5 part, for example, 0.1 part, 0.2 part, 0.3 part, 0.4 part, 0.5 part and the like.
Preferably, the hair loss preventing and hair growing composition further comprises an angelica extract, a pricklyash peel extract, a radix sophorae flavescentis extract and a fructus forsythiae extract.
In the invention, the angelica extract, the pricklyash peel extract, the radix sophorae flavescentis extract and the fructus forsythiae extract are additionally added into the hair loss prevention and hair growth composition, so that the capabilities of balancing scalp grease, reducing dandruff, sterilizing and inhibiting bacteria and preventing hair loss are further improved.
Wherein, the main component of the angelica extract is angelica polysaccharide, which gives nutrition to scalp and improves the nutrition transportation of scalp, thereby recovering the cell functions of scalp epidermis, dermis and subcutaneous tissue and realizing the healthy and omnibearing care of scalp; the Chinese prickly ash fruit extract has pungent and warm taste, warms middle energizer to relieve pain, kills parasites and relieves itching; matrine and kurarinone in the kuh-seng extract are used as antibacterial components in the hair-growing agent; the forsythol, forsythoside A and other components contained in the forsythia extract have good antibacterial effects, and comprise streptococcus, cholera solitary fungus, typhoid bacillus, diphtheria bacillus, escherichia coli, shigella dysenteriae, staphylococcus and the like.
Preferably, the anti-hair loss hair-growing composition comprises the following components in parts by weight: 0.01-1 part of liquiritin, 1-3 parts of Boerduo leaf extract, 0.01-2 parts of oleanolic acid, 0.1-0.5 part of stilbene glucoside, 5-10 parts of angelica extract, 1-5 parts of Chinese prickly ash fruit extract, 3-8 parts of radix sophorae flavescentis extract and 3-8 parts of fructus forsythiae extract.
In the hair loss preventing and hair growing composition, the content of the angelica sinensis extract is 5-10 parts, for example, 5 parts, 5.5 parts, 6 parts, 6.5 parts, 7 parts, 7.5 parts, 8 parts, 8.5 parts, 9 parts, 9.5 parts, 10 parts and the like can be adopted.
The content of the pricklyash peel extract in the hair-growing composition is 1-5 parts, for example, 1 part, 1.5 parts, 2 parts, 2.5 parts, 3 parts, 3.5 parts, 4 parts, 4.5 parts, 5 parts and the like.
In the hair loss preventing and hair growing composition, the content of the kuh-seng extracting solution is 3-8 parts, for example, 3 parts, 3.5 parts, 4 parts, 4.5 parts, 5 parts, 5.5 parts, 6 parts, 6.5 parts, 7 parts, 7.5 parts, 8 parts and the like can be adopted.
In the hair-loss prevention and hair-growing composition, the content of the weeping forsythiae capsule extract is 3-8 parts, and can be 3 parts, 3.5 parts, 4 parts, 4.5 parts, 5 parts, 5.5 parts, 6 parts, 6.5 parts, 7 parts, 7.5 parts, 8 parts and the like.
Preferably, the hair-growing composition further comprises a solvent, wherein the solvent is a mixed solution of alcohol and water.
Preferably, the mass ratio of the alcohol to the water is 1 (1-2), which may be, for example, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, etc.
Preferably, the alcohol comprises any one or a combination of at least two of ethanol, isopentyl glycol, octyl glycol, propylene glycol, butylene glycol, hexylene glycol or glycerin, preferably butylene glycol and/or hexylene glycol.
Preferably, the anti-hair loss hair-growing composition comprises the following components in percentage by mass: 0.01-1% of glycyrrhizin, 1-3% of Boerduo leaf extract, 0.01-2% of oleanolic acid, 0.1-0.5% of stilbene glucoside, 5-10% of Chinese angelica extract, 1-5% of pricklyash peel extract, 3-8% of radix sophorae flavescentis extract, 3-8% of fructus forsythiae extract, 10-30% of butanediol, 10-30% of hexanediol and the balance of water.
The content of glycyrrhizin is 0.01-1% based on 100% of the total weight of the hair-growth composition, and may be, for example, 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, etc.
The content of the Boerdosis leaf extract is 1-3% based on 100% of the total weight of the hair-growing composition, and may be 1%, 1.2%, 1.4%, 1.6%, 1.8%, 2%, 2.2%, 2.4%, 2.6%, 2.8%, 3%, etc.
The oleanolic acid content is 0.01-2% based on 100% total weight of the hair-growing composition, and may be, for example, 0.01%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.6%, 0.8%, 1%, 1.2%, 1.4%, 1.6%, 1.8%, 2%, etc.
The stilbene glycoside content is 0.1-0.5% based on 100% of the total weight of the hair-growth composition, and may be, for example, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, etc.
The content of the radix Angelicae sinensis extract is 5-10%, for example 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, etc., based on 100% of the total weight of the hair growth promoting composition.
The content of fructus Zanthoxyli extract is 1-5%, for example, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, etc. based on 100% total weight of the hair growth promoting composition.
The content of radix Sophorae Flavescentis extract is 3-8%, for example 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, etc. based on 100% total weight of the hair growth promoting composition.
The fructus forsythiae extract is 3-8% based on 100% of the total weight of the hair-caring composition, and can be 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, etc.
The content of butanediol is 10-30% based on 100% of the total weight of the hair-growth composition, and may be, for example, 10%, 12%, 14%, 16%, 18%, 20%, 22%, 24%, 26%, 28%, 30%, etc.
The content of hexanediol is 10-30% based on 100% of the total weight of the hair-growing composition, and may be, for example, 10%, 12%, 14%, 16%, 18%, 20%, 22%, 24%, 26%, 28%, 30%, etc.
In a second aspect, the present invention provides a method for preparing the hair-setting preventing composition as in the first aspect, comprising: dissolving liquiritin, boerduo leaf extract, oleanolic acid and stilbene glucoside, and filtering to obtain the hair-caring composition.
Preferably, the preparation method of the hair-loss preventing hair-growing composition comprises the following steps: dissolving liquiritin, boerduo leaf extract, oleanolic acid, stilbene glucoside, radix Angelicae sinensis extract, fructus Zanthoxyli extract, radix Sophorae Flavescentis extract and fructus forsythiae extract in mixed solution of alcohol and water, mixing, stirring, and filtering to obtain the hair loss preventing composition.
In a third aspect, the present invention provides the use of an anti-hair loss hair-growing composition according to the first aspect for the preparation of a cosmetic.
In a fourth aspect, the present invention provides a cosmetic comprising an anti-hair-growing composition according to the first aspect.
Preferably, the cosmetic comprises any one or a combination of at least two of shampoo, essence or conditioner.
Preferably, the anti-hair loss and hair growth composition is added in an amount of 1-20% by weight, for example, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, etc., preferably 12-18%, further preferably 15% by weight, based on the total weight of the cosmetic.
Compared with the prior art, the invention has the following beneficial effects:
(1) The anti-hair loss composition provided by the invention effectively inhibits the activity of 5 alpha-reductase through the synergistic combination of four effective components of glycyrrhizin, the Boedo leaf extract, oleanolic acid and stilbene glucoside, so that the problems that the high-activity DHT is combined with an androgen receptor and then acts on hair follicles to generate rapid entry in a hair withdrawal period, micromation, hair loss and the like are avoided to a great extent;
(2) The anti-hair loss composition can inhibit the expression of IL-1 beta and TNF-alpha inflammatory factors in skin hair follicles by the synergistic combination of four active ingredients of liquiritin, boerduo tree leaf extract, oleanolic acid and stilbene glucoside, so that the purposes of preventing hair loss and promoting hair growth are finally achieved;
(3) The hair loss prevention and hair care composition can prevent and improve the condition of hair loss, keep the cortex sanitary, ventilate, not greasy, astringe excessively hypertrophic sebaceous glands, has the effects of reducing sebum secretion and promoting blood circulation, and has the effects of cleaning and stimulating the hair which is shed.
Detailed Description
The technical scheme of the invention is further described by the following specific embodiments. It will be apparent to those skilled in the art that the examples are merely to aid in understanding the invention and are not to be construed as a specific limitation thereof.
The sources of the components of the following examples and comparative examples are shown below:
in the embodiment, the mass ratio of the angelica extract is (5): 6) =5:3
Example 1
The embodiment provides an anti-hair-loss hair-growing composition, which comprises the following components in percentage by mass based on the total mass of the anti-hair-loss hair-growing composition of 100 percent:
the preparation method of the hair-loss prevention hair-growing composition comprises the following steps: dissolving liquiritin, boerduo leaf extract, oleanolic acid, stilbene glucoside, radix Angelicae sinensis extract, fructus Zanthoxyli extract, radix Sophorae Flavescentis extract and fructus forsythiae extract in the above mixed solution of alcohol and water, mixing, stirring, and filtering to obtain the hair loss preventing hair growth promoting composition.
Example 2
The embodiment provides an anti-hair-loss hair-growing composition, which comprises the following components in percentage by mass based on the total mass of the anti-hair-loss hair-growing composition of 100 percent:
the preparation method of the hair-loss prevention hair-growing composition comprises the following steps: dissolving liquiritin, boerduo leaf extract, oleanolic acid, stilbene glucoside, radix Angelicae sinensis extract, fructus Zanthoxyli extract, radix Sophorae Flavescentis extract and fructus forsythiae extract in the above mixed solution of alcohol and water, mixing, stirring, and filtering to obtain the hair loss preventing hair growth promoting composition.
Example 3
The embodiment provides an anti-hair-loss hair-growing composition, which comprises the following components in percentage by mass based on the total mass of the anti-hair-loss hair-growing composition of 100 percent:
the preparation method of the hair-loss prevention hair-growing composition comprises the following steps: dissolving liquiritin, boerduo leaf extract, oleanolic acid, stilbene glucoside, radix Angelicae sinensis extract, fructus Zanthoxyli extract, radix Sophorae Flavescentis extract and fructus forsythiae extract in the above mixed solution of alcohol and water, mixing, stirring, and filtering to obtain the hair loss preventing hair growth promoting composition.
Examples 4 to 7
Examples 4-7 provide four anti-hair loss hair-growing compositions, differing from example 1 in the absence of any one of the angelica sinensis extract, the pricklyash peel extract, the kuh-seng extract and the weeping forsythia extract, respectively, and the preparation method is the same as example 1; the anti-hair loss hair-growing composition comprises the following components in percentage by mass:
comparative example 1
Comparative example 1 provides an anti-hair loss composition, which is different from example 1 in that no glycyrrhizin, boldo leaf extract, oleanolic acid and stilbene glycoside are added, and the preparation method is the same as example 1; the anti-hair loss hair-growing composition comprises the following components in percentage by mass:
comparative examples 2 to 5
Comparative examples 2 to 5 provide four hair-growing compositions differing from example 1 in that only one component of glycyrrhizin, boldo leaf extract, oleanolic acid and stilbene glycoside is contained; the anti-hair loss hair-growing composition comprises the following components in percentage by mass:
comparative examples 6 to 8
Comparative examples 6-8 provide three anti-hair-loss hair-growing compositions, differing from example 1 in that only two components of liquiritin, boldo leaf extract and oleanolic acid were compounded; the anti-hair loss hair-growing composition comprises the following components in percentage by mass:
comparative examples 9 to 11
Comparative examples 9-11 provide four anti-hair-loss hair-growing compositions, differing from example 1 in the absence of an example of one of glycyrrhizin, boldo leaf extract, oleanolic acid; the anti-hair loss hair-growing composition comprises the following components in percentage by mass:
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test example 1
5 alpha reductase Activity inhibition Rate test
Test sample: the anti-hair loss hair-growing compositions prepared in examples 1 to 7 and comparative examples 1 to 11;
the testing method comprises the following steps: the method specifically comprises the following steps:
(1) Preparation of 5a reduction crude enzyme:
the male SD rat is killed by 10% chloral hydrate abdominal cavity excessive injection, liver tissue is rapidly dissected and removed, redundant blood is washed away by normal saline, phosphate buffer (pH=7.4) is added according to the mass ratio of 1:5, magnetic beads are added into a homogenizer for homogenization, the homogenization is taken out and centrifuged for 10min at 4 ℃ 10000r/min on a refrigerated centrifuge, the supernatant is taken out and centrifuged once again, the supernatant is taken out as crude enzyme liquid, and the crude enzyme liquid is frozen at-20 ℃ for standby.
(2) Protein content determination:
the protein concentration in the crude enzyme solution is determined by using a protein quantitative kit, and the protein standard is diluted to a mass concentration of 0, 10, 20, 40, 80, 120, 160, 200 mug/mL series of standard protein solutions according to the experimental method provided by the kit. 200 mu L of G250 staining solution is added into each series of standard solutions, after being uniformly mixed, the mixture is placed at room temperature for 3-5min, the absorbance of the mixture is detected at the wavelength of 595nm by using an enzyme-labeled instrument, and a standard curve is drawn by taking the absorbance (Y) of protein as an ordinate and the mass concentration (X, mg/mL) as an abscissa. The crude enzyme protein mass concentration was calculated by the protein standard curve equation.
3) Assay for 5 alpha reductase Activity inhibition Activity
The total reaction volume of the 5 alpha reductase catalytic reaction is 200 mu L,120mg/mL liver homogenate, 5mg/mL NADPH, 0.2mg/mL testosterone and samples with different concentrations to be tested.
Blank group assay: to a 96-well plate, 120. Mu.L of liver homogenate, 60. Mu.L of 5mg/mLNADPH, 10. Mu.L of testosterone (0.2 mg/mL), 10. Mu.L of phosphate buffer (pH 7.4) were added, the mixture was thoroughly mixed, absorbance A0 was measured, and after incubation at 37℃for 60 minutes, absorbance A1 was measured to obtain the NADPH change value DeltaA 0.
Sample group assay: respectively preparing to-be-detected liquid (2, 5, 10, 20, 30, 40 mg/mL) of to-be-detected samples with different concentrations, and respectively carrying out reaction to obtain the NADPH variation value delta An.
Positive group determination, namely judging whether the activity of the enzyme is inhibited or not by using finasteride as a positive control drug according to the method, and primarily calculating the inhibition rate.
The calculation formula is I% = (Δan- Δao)/[ Δao×100%. Experiments were repeated 3 times, 3 replicates each.
The specific test results are shown in Table 1
(wherein, the anti-hair-growing composition is represented by a code number C4, DPO represents diaminopyrimidine oxide, and dilution is carried out with water to 100%, "-" represents that the item was not tested):
TABLE 1
The test results in table 1 show that the anti-hair loss composition has an inhibition rate of more than 180% on 5 alpha reductase activity, and fully shows that the anti-hair loss composition can effectively penetrate into scalp and hair follicles through synergistic compounding of four effective components of glycyrrhizin, a Boldo leaf extract, oleanolic acid and stilbene glycoside, so that 5 alpha-reductase activity is effectively inhibited, 5 alpha-reductase is greatly prevented from catalyzing testosterone to be converted into DHT, and then the problems of rapid entry into a retrogradation period, micromation, hair loss and the like caused by acting on hair follicles after the high-activity DHT is combined with androgen receptor are avoided.
In addition, in a dilution experiment, the hair-loss prevention hair-growing composition disclosed by the invention is suitable for being used in cosmetics, and the total mass of the cosmetics is about 15%, so that the inhibition rate of the activity of 5 alpha reductase can be improved, and the defects and waste caused by excessive addition are avoided. Meanwhile, it was not difficult to find that the addition of ethanol and/or diaminopyrimidine oxide further improved the inhibition of 5α reductase activity by the composition at low concentrations (5-15% of the C4 composition added).
Test example 2
Inflammatory factor inhibition test
Test sample: the anti-hair loss hair-growing compositions prepared in examples 1 to 7 and comparative examples 1 to 11;
the testing method comprises the following steps: human immortalized keratinocytes (HaCaT cells) in the dosing group and the blank group were stimulated with Lipopolysaccharide (LPS), respectively, and protein expression of the relevant inflammatory factors interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) in the HaCaT cell supernatants of the dosing group pretreatment group and the control group was detected by ELISA.
HaCaT cells were routinely cultured in DMEM complete medium (1% diabody: penicillin 100U/mL and streptomycin 100. Mu.g/mL added) containing 10% (volume fraction) fetal bovine serum at 37℃in a 5% (volume fraction) CO2 saturated humidity incubator with liquid changes every 2-3 days for 3-7 days. Taking HaCaT cells in logarithmic growth phase, digesting the cells for 10min by using 0.25% trypsin-EDTA, washing the cells, adding a complete culture medium into the cell sediment, and gently beating into a cell suspension. The cell concentration was adjusted to 4X 10 with fresh culture medium 4 Cells were inoculated onto 24 well plates, incubated in a cell incubator for 24h, and LPS was added after the cells had adhered to about 60% -70%. The blank group, the lipopolysaccharide LPS 20 mug/mL stimulation 24h group, the different administration groups are pretreated for 24h+LPS 20 mug/mL stimulation 24h group, each group is provided with 5 compound holes, after the stimulation is finished, cell supernatant of each hole is sucked and split charged into an EP tube of 1.5mL, and the cell supernatant can be used immediately or frozen at-20 ℃ for standby. The supernatants were tested for the expression levels of TNF- α and IL-1β using ELISA kit (Jiangsu enzyme-labeled Biotech Co., ltd.).
The specific test results are shown in table 2:
TABLE 2
As shown in the test data of Table 2, the anti-hair loss composition has good inhibition effect on the generation of inflammatory factors, the content of inflammatory factors IL-beta is reduced to below 125 after the composition is used, the content of inflammatory factors TNF-alpha is reduced to below 126, and the synergistic combination of four effective components of glycyrrhizin, podol leaf extract, oleanolic acid and stilbene glucoside is fully shown, and meanwhile, the anti-inflammatory effect is achieved by inhibiting the IL-1 beta and TNF-alpha 2 inflammatory factors which are over-expressed in skin hair follicles.
Test example 3
Mouse experiment
Test sample: the anti-hair loss hair-growing compositions prepared in examples 1 to 7 and comparative examples 1 to 11;
the testing method comprises the following steps: c57 mice (male, 5 weeks old) were kept in an environment of 23+ -2deg.C and 54+ -10% humidity, and were allowed to eat and drink water. Except for the blank group, the backs of the mice are selected to be injected with medicines for subcutaneous injection, testosterone propionate is injected for 5 mg/kg.d, the medicine is injected after 4 weeks of modeling success, physiological saline is smeared on the model group and the blank group, 5% minoxidil tincture is smeared on the minoxidil group for 0.2mL/d, and 15% of composition aqueous solution is smeared on the sample group for 1 time every day for 4 weeks.
(1) Final hair/vellus hair value ratio: after the observation is finished at the 8 th week of the experiment, the mice are sacrificed, the skin of the back alopecia area of each group of mice is flatly attached to a fixed plate after about 1cm multiplied by 1cm, paraffin section dewaxing, hematoxylin-eosin staining and dehydration sealing experiment operation are carried out, under the field of 100 times of microscope, morphological changes of hair follicles of the mice are observed under the condition of 3 non-overlapping fields on average, the lengths and the sizes of the hair follicles are measured by utilizing Image-Pro Plus, and the ratio (A/T) of the number of hair follicles in the growing period to the number of hair follicles in the resting period is counted;
(2) ELISA determination of Dihydrotestosterone (DHT) and 5α reductase content in skin tissue: preparing tissue homogenate from tissues obtained by dissecting each group of mice, measuring the DHT and 5 alpha-reductase content in each group of samples by using an Elisa kit according to the method on an ELISA kit specification, recording data, and obtaining the average value by using Excel;
(3) Immunohistochemical determination of TGF- β1 and TNF- α content in skin: each group of mice is obtained by taking 2.0cm multiplied by 0.8cm long strip skin, placing the long strip skin in normal saline, fully rinsing residual blood, spreading the long strip skin on filter paper, airing the long strip skin at room temperature, wrapping the long strip skin on tinfoil paper, performing immunohistochemical detection to determine the contents of TGF-beta 1 and TNF-alpha in the skin of the mice according to the using method of Anti-TGFB1 and TNF-alpha, recording data, using Excel, and obtaining the average value of the data;
the specific test results are shown in table 3: (wherein, the anti-hair-growing composition is represented by the code C4, "-" represents that the item was not tested):
TABLE 3 Table 3
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As can be seen from the test data in Table 3, after the anti-alopecia composition of the present invention is used, the ratio of the final hair/vellus hair value of the HE staining result of the back test tissue of the mice is obviously increased to more than 4; ELISA measures the significant decrease in the content of Dihydrotestosterone (DHT) and 5α reductase in skin tissue, and immunohistochemistry measures the significant decrease in the content of TGF- β1 and TNF- α in skin.
Therefore, the synergistic combination of four effective components of the liquiritin, the Boerduo leaf extract, the oleanolic acid and the stilbene glucoside can effectively penetrate into scalp and hair follicles, effectively inhibit 5 alpha-reductase and improve hair follicle atrophy caused by DHT; simultaneously inhibit the over-expressed IL-1 beta and TNF-alpha inflammatory factors caused by the breeding of anaerobic acne bacillus in skin hair follicles, can realize the simultaneous treatment of both symptoms and root causes to play roles in controlling oil and resisting inflammation, radically improve hair follicle microcirculation, stimulate cell metabolism and radically solve the problem of alopecia.
The applicant states that the present invention is illustrated by the above examples as well as methods of making and using the same, but the present invention is not limited to, i.e., does not mean that the present invention must be practiced in dependence upon the above examples. It should be apparent to those skilled in the art that any modification of the present invention, equivalent substitution of raw materials for the product of the present invention, addition of auxiliary components, selection of specific modes, etc., falls within the scope of the present invention and the scope of disclosure.
Claims (15)
1. The hair-loss prevention hair-growing composition is characterized by comprising the following components in parts by weight: 0.01-1 part of liquiritin, 1-3 parts of Boerduo leaf extract, 0.01-2 parts of oleanolic acid and 0.1-0.5 part of stilbene glucoside;
the glycyrrhizin is derived from a licorice root extract, and the mass percentage of the glycyrrhizin in the licorice root extract is more than or equal to 50%;
the stilbene glucoside is derived from a polygonum multiflorum extract, and the mass percentage of the stilbene glucoside in the polygonum multiflorum extract is more than or equal to 50%.
2. The hair-loss preventing and hair-growing composition according to claim 1, further comprising an angelica extract, a pricklyash peel extract, a kuh-seng extract and a weeping forsythia extract.
3. The anti-hair-extension composition according to claim 1, wherein the anti-hair-extension composition comprises, in parts by weight: 0.01-1 part of liquiritin, 1-3 parts of Boerduo leaf extract, 0.01-2 parts of oleanolic acid, 0.1-0.5 part of stilbene glucoside, 5-10 parts of angelica extract, 1-5 parts of Chinese prickly ash fruit extract, 3-8 parts of radix sophorae flavescentis extract and 3-8 parts of fructus forsythiae extract.
4. The anti-hair-growing composition according to claim 1, further comprising a solvent, which is a mixed solution of alcohol and water.
5. The hair-setting and hair-growing composition according to claim 4, wherein the mass ratio of the alcohol to the water is 1 (1-2).
6. The anti-hair-setting composition according to claim 4, wherein the alcohol comprises any one or a combination of at least two of ethanol, isopentyl glycol, octanyl glycol, propylene glycol, butylene glycol, hexylene glycol, or glycerin.
7. The anti-hair-setting composition according to claim 4, wherein the alcohol is butanediol and/or hexanediol.
8. The hair-loss prevention and hair-growth composition according to claim 1, which comprises the following components in percentage by mass: 0.01-1% of glycyrrhizin, 1-3% of Boerduo leaf extract, 0.01-2% of oleanolic acid, 0.1-0.5% of stilbene glucoside, 5-10% of Chinese angelica extract, 1-5% of pricklyash peel extract, 3-8% of radix sophorae flavescentis extract, 3-8% of fructus forsythiae extract, 10-30% of butanediol, 10-30% of hexanediol and the balance of water.
9. The method for preparing an anti-hair-extension composition according to claims 1 to 8, wherein the method for preparing the anti-hair-extension composition comprises: dissolving liquiritin, boerduo leaf extract, oleanolic acid, stilbene glucoside, radix Angelicae sinensis extract, fructus Zanthoxyli extract, radix Sophorae Flavescentis extract and fructus forsythiae extract in mixed solution of alcohol and water, mixing, stirring, and filtering to obtain the hair loss preventing composition.
10. Use of the anti-hair loss hair-growing composition according to any one of claims 1 to 8 in the preparation of a cosmetic.
11. A cosmetic product characterized in that it comprises the hair-loss preventing hair-growing composition according to any one of claims 1 to 8.
12. The cosmetic product of claim 11, wherein the cosmetic product comprises any one or a combination of at least two of a shampoo, a cream, a serum, or a conditioner.
13. The cosmetic according to claim 11, wherein the anti-hair loss composition is added in an amount of 1 to 20% by weight of the total mass of the cosmetic.
14. The cosmetic according to claim 11, wherein the anti-hair loss composition is added in an amount of 12 to 18% by weight of the total mass of the cosmetic.
15. The cosmetic according to claim 11, wherein the anti-hair loss composition is added in an amount of 15% of the total mass of the cosmetic.
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| CN115212144B (en) * | 2022-07-24 | 2023-06-23 | 广州市比柔生物科技有限公司 | Stable and effective antibacterial itching-relieving alopecia-preventing composition and preparation and application thereof |
| WO2024052756A1 (en) * | 2022-09-09 | 2024-03-14 | Crodarom | Use of an extract of sichuan pepper for a skin treatment and a composition suitable for this use. |
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