CN119185115A

CN119185115A - Application of Akebia extract in cosmetics

Info

Publication number: CN119185115A
Application number: CN202310756818.5A
Authority: CN
Inventors: 孙懿; 祝乐; 赵亚; 贾海东
Original assignee: Shanghai Jahwa United Co Ltd
Current assignee: Shanghai Jahwa United Co Ltd
Priority date: 2023-06-26
Filing date: 2023-06-26
Publication date: 2024-12-27

Abstract

The application of Akebia extract in skin oil control and/or hair loss prevention, wherein the Akebia extract is prepared by solvent extraction, and the skin oil control and/or hair loss prevention is achieved by inhibiting 5α-reductase. The present invention also provides the application of Akebia extract in the preparation of a skin topical agent with skin oil control and/or hair loss prevention effects.

Description

Application of akebia stem extract in cosmetics

Technical Field

The invention belongs to the field of phytochemistry, and particularly relates to a akebia stem extract and application thereof in external preparations or cosmetics for skin.

Background

Androgenetic alopecia (AGA) is mainly manifested by shortening of the anagen phase of hair follicles and miniaturization of terminal hair follicles. Experiments prove that androgens play a critical role in the pathogenesis of AGA. Testosterone is the major androgen in the body and is converted to Dihydrotestosterone (DHT) by 5-alpha reductase, which is 5 times more compatible with the androgen receptor than testosterone, and can cause a final hair to vellus conversion ultimately leading to hair loss. Current treatment modalities include both drug and non-drug therapies, such as 5-alpha reductase inhibitors, platelet rich plasma and hair transplantation, and the like.

The 5-alpha reductase inhibitor can effectively inhibit testosterone conversion to DHT, thereby reducing DHT of scalp in alopecia area and promoting hair growth.

Wu Chuanling and the like prove that the lotus plumule extract has 5-alpha reductase inhibition activity through establishing an in-vitro 5-alpha reductase screening model, deng Guiqiu and the like establish 5-alpha reductase inhibition in-vitro models through a high performance liquid chromatography, researches prove that the safflower extract has a certain inhibition effect on type I and type II 5-alpha reductase, zhao Jideng determines the 5-alpha reductase inhibition activity of the nettle extract through a high performance liquid chromatography, researches show that the n-butyl alcohol part extracted from the nettle whole herb has obvious 5-alpha reductase inhibition activity, kazuya and the like discover that pepper, schisandra chinensis, kudzuvine flower, ginsenoside and the like have a certain inhibition effect on 5-alpha reductase activity through establishing an in-vitro enzyme activity system, and Zhang Bei and the like screen out the biota extract from 9 traditional Chinese medicines with the effects of tonifying kidney liver, benefiting essence and blood and cooling blood. The screening system of 5-alpha reductase in vitro activity is used for screening the effective monomers with inhibitory activity, such as total glycosides of rhizoma anemarrhenae, forsythin and lupulone, yang Xiaoqing and the like, through the research on the inhibitory activity of 5-alpha reductase of methyl lotus plumule and isoliensinine, the results show that the total glycosides of rhizoma anemarrhenae, forsythin and lupulone have stronger inhibitory activity, have application prospect, ju Jiang and the like, through the research on the influence of berberine hydrochloride and baicalin on the expression of 5-alpha reductase, the berberine hydrochloride is found to possibly antagonize the stimulation of dihydrotestosterone on the activity of human sebaceous gland cells through the inhibition of the expression of 5-alpha reductase.

Akebia quinata is a dried rattan of the family Akebiaceae plant Akebia quinata Akebia quinata (thunder.) Decne, akebia trifoliata Akebia trifoliata (thunder.) Koidz, or Akebia quinata Akebia trifoliata (thunder.) Koidz.

KR100816264 discloses an analgesic skin cosmetic composition (Cosmetic Composition for Abirritating Skin Containing Extract of Akebia quinata as Active Ingredient). containing akebia stem extract as an active ingredient, and the patent reports that akebia stem has an effect of relieving irritation.

KR20150045420 discloses a composition comprising akebia stem extract for blocking AGEs production (Composition for blocking AGEs production comprising Akebia quinata extracts). The patent reports an anti-glycation effect of akebia stem, which can be used for anti-aging products.

KR102036826 discloses a composition for preventing or treating atopic dermatitis comprising akebia stem extract or a part thereof as an active ingredient (Composition for preventing or treating atopic dermatitis comprising Akebia quinata extracts or fractions thereof as an active ingredient).

KR20140071716 discloses a cosmetic composition (Cosmetic composition comprising the extract of akebia quinata AS ACTIVE INGREDIENT) comprising akebia stem extract as an active ingredient. The patent reports that akebia stem has anti-inflammatory, whitening and moisturizing effects.

KR101427462 discloses a complex natural preservative from forsythia, motherwort, hawthorns and akebia stem, and cosmetic composition (Composite Natural Antiseptics From Forsythia suspensa Vahl,Leonurus sibiricus L.,Crataegi fructus and Akebia quinata Decaisne,Cosmetic Compositions Containing Them). containing them, which reports that the akebia stem composition has antibacterial and preservative effects.

CN201711486685.5 discloses a Chinese herbal medicine composition for treating alopecia. The patent reports a Chinese herbal composition containing akebia stem.

However, none of these prior art documents find application of akebia stem single drug as skin oil control and/or anti-hair loss efficacy ingredient in cosmetics.

Disclosure of Invention

In one aspect, the invention provides application of akebia stem extract in skin oil control and/or alopecia prevention, wherein the akebia stem extract is prepared by a solvent extraction method.

In a preferred embodiment, the akebia stem extract is prepared by extraction by decoction using water as a solvent.

In a preferred embodiment, the akebia stem extract is used at a concentration of 0.001 to 1mg/mL, more preferably 0.007mg/mL to 0.5mg/mL.

In a preferred embodiment, the skin oil control and/or hair loss prevention is achieved by inhibiting 5α -reductase.

In another aspect, the invention also relates to application of the akebia stem extract in preparing external skin preparations with skin oil control and/or alopecia prevention functions, wherein the akebia stem extract is prepared by a solvent extraction method.

In a preferred embodiment, the weight percentage of the akebia stem extract in the skin external agent is 0.0005 to 10 wt%, preferably 0.000731 to 5wt%, more preferably 0.003655 to 1 wt%, most preferably 0.00731 to 0.5 wt%.

In a preferred embodiment, the skin external preparation is selected from the group consisting of face cream, milky lotion, jelly, face toilet, essence, pack, eye cream, aerosol cleansing foam, spray, body wash, and facial cleanser.

Brief description of the drawings

Figure 1 shows the chromatographic results of testosterone standards.

Fig. 2 shows a linear equation for testosterone content measurement.

Detailed Description

The invention discovers that the akebia stem extract has the effect of inhibiting 5-alpha reductase and can be used as a single efficacy additive to be added into skin care products for assisting in relieving relevant problems such as scalp, alopecia prevention and the like.

In order to provide a more concise description, some quantitative representations presented herein are not modified by the term "about". It will be understood that each quantity given herein is intended to refer to an actual given value, whether or not the term "about" is explicitly used, and is also intended to refer to approximations of such given values, including approximations of such given values resulting from experimental and/or measurement conditions, as reasonably deduced by one of ordinary skill in the art.

To provide a more concise description, some quantitative expressions herein are recited as a range from about X to about Y. It should be understood that when a range is recited, the range is not limited to the recited upper and lower limits, but rather, includes the entire range of about X to about Y amounts or any amount therebetween.

Akebia stem extract

The invention discovers for the first time that the akebia stem extract has the effect of inhibiting 5-alpha reductase, and the akebia stem single medicine has the effects of controlling oil on skin and/or preventing alopecia. Therefore, the akebia stem extract can be used as a single efficacy additive to be added into skin care products to help relieve relevant problems such as scalp, alopecia prevention and the like.

Specifically, the present invention finds that akebia stem extract has 5α -reductase inhibitory effect. Therefore, the akebia stem extract disclosed by the invention can help to relieve scalp, has the effects of controlling oil on skin and/or preventing alopecia, and can be used as an efficacy additive to be added into an external skin preparation for controlling oil on skin and/or preventing alopecia.

5 Alpha-reductase is an important enzyme in the metabolic process of androgens, and can convert testosterone into dihydrotestosterone with stronger activity, and the dihydrotestosterone can widely participate in the metabolism of androgens in vivo, thereby playing an important physiological role. Abnormal expression of 5α -reductase in vivo may lead to abnormal levels of local tissues or systemic hormones, thereby leading to the occurrence of various diseases such as acne, androgenic alopecia, etc.

In sebaceous gland cells, androgens can be converted into more active dihydrotestosterone by 5 alpha-reductase, and sebum secretion is further stimulated, so that the capability of inducing acne is enhanced. Therefore, inhibiting the activity of 5α -reductase can also control the secretion of skin oil and effectively alleviate the production of acne.

Furthermore, hormonal alopecia is positively correlated with the content of local androgens, especially dihydrotestosterone, and 5α -reductase is a key enzyme for testosterone conversion to dihydrotestosterone. Thus, inhibition of 5α -reductase activity is effective in alleviating seborrheic alopecia.

In some embodiments, the akebia stem is extracted using a thermal extraction method. For example, extraction may be performed by decoction. In some embodiments, the extraction solvent is water (e.g., deionized water). In some embodiments, the extraction process includes an alcohol precipitation step. For example, 95% ethanol is used for ethanol precipitation to a final concentration of 70%. In some embodiments, the extraction process includes a filtration step. In some embodiments, the extraction process further comprises a concentration step. For example, to a solids content of 5 to 10% by weight.

In some embodiments, the akebia stem extract is used at a concentration of 0.001mg/mL to 1mg/mL. A preferred concentration is used in the range of 0.007mg/mL to 1mg/mL. More preferably, the concentration is used in the range of 0.007mg/mL to 0.5mg/mL. More preferably, the concentration is used in the range of 0.007mg/mL to 0.1mg/mL. Most preferably, the concentration is 0.00731mg/mL-0.0731mg/mL.

External preparation for skin containing Akebia quinata extract

The akebia stem extract can be used as an efficacy additive to be applied to external skin preparations. In some embodiments, the skin external is selected from the group consisting of a cleanser, a lotion, an emulsion, a cream, a jelly, and a mask. Different amounts are added according to the different types of formulations.

In another aspect, there is provided a skin external preparation comprising the akebia stem extract prepared according to the method of the present invention and an excipient acceptable in the cosmetic field.

The external skin preparation is a general concept of all ingredients commonly used outside the skin, and may be, for example, a cosmetic composition or a pharmaceutical composition. The cosmetic composition may be basic cosmetic, facial makeup cosmetic, body cosmetic, hair care cosmetic, etc., and its dosage form is not particularly limited and may be reasonably selected according to different purposes. The cosmetic composition also contains various cosmetically acceptable medium or matrix excipients depending on dosage form and purpose.

The cosmetically, dermatologically or pharmaceutically acceptable excipients that may be used in the skin external composition of the present invention are in the form of an aqueous phase, an oil phase, a gel, a wax-in-water emulsion, an oil-in-water emulsion or a water-in-oil emulsion. The aqueous phase is a mixture of one or more water-soluble or dispersible components, which may be liquid, semi-solid or solid at room temperature (25 ℃). Excipients include or may be in the form of suspensions, dispersions or solutions in water or water-alcohol excipients, which may contain a thickening agent or gelling agent. The skilled person can choose the appropriate product form, based on knowledge known to the person skilled in the art, in which the components are comprised.

The composition may comprise an aqueous phase which may contain water or a mixture of water and at least one hydrophilic organic solvent such as alcohols, in particular straight or branched chain lower monohydric alcohols containing from 2 to 5 carbon atoms, such as ethanol or propanol, polyols, such as propylene glycol, sorbitol, glycerol, panthenol or polyethylene glycol and mixtures thereof.

When the composition of the invention is in the form of an emulsion, the composition may optionally further comprise a surfactant.

The composition may also comprise film-forming polymers such as polyurethanes, polyacrylic homo-or copolymers, polyesters, hydrocarbon-based resins and/or silicone resins. The polymer may be dissolved or dispersed in a cosmetically acceptable vehicle and optionally combined with a plasticizer.

The compositions of the present invention may also comprise an oil phase containing oil-soluble or oil-dispersible components that are liquid at room temperature (25 ℃) and/or substances that are oily or waxy at room temperature, such as waxes, semi-solids, gums, and mixtures thereof. The oil phase may also contain an organic solvent.

Suitable oily substances which are generally liquid at room temperature include hydrocarbon-based oils of animal origin, such as perhydro squalene, hydrocarbon-based vegetable oils, such as triglycerides of liquid C4-10 fatty acids, for example heptanoic acid or caprylic triglycerides, or oils, for example sunflower oil, corn oil, soybean oil, grapeseed oil, castor oil, avocado oil, caprylic/capric triglycerides, jojoba oil, linear or branched hydrocarbons of mineral or synthetic origin, for example liquid paraffin and its derivatives, vaseline, synthetic esters and ethers, in particular esters of fatty alcohols, for example isopropyl myristate, 2-ethylhexyl palmitate, 2-octyldodecyl stearate, isostearyl isostearate, hydroxylated esters, for example isostearyl lactate, octyl hydroxystearate, octyldodecyl hydroxystearate, heptanoic acid esters of fatty alcohols, caprylic and capric acid esters, polyol esters, for example propylene glycol dicaprylate, neopentyl glycol diheptanoic acid esters, diethylene glycol diisononanoic acid esters and pentaerythritol esters, fatty alcohols containing C12-26, for example 2-octyldodecyl palmitate, 2-octyldodecyl stearate, isostearyl silicone, methyl silicone, 2-methyl-undecyl-phenyl-containing silicone, or a cyclic silicone oil, or a mixture thereof, and optionally phenyl-containing silicone, and optionally methyl-or a mixture thereof.

The composition of the present invention may further comprise any component commonly used in the cosmetic field. These components include preservatives, aqueous phase thickeners (extract biopolymers, synthetic polymers) and fatty phase thickeners, fragrances, hydrophilic and lipophilic active agents, and mixtures thereof.

The compositions of the present invention may also comprise additional particulate phases which may be pigments and/or pearlizing agents and/or fillers used in cosmetic compositions.

Pigments may be present in the composition, suitable inorganic pigments include titanium oxide, zirconium oxide and cerium oxide, zinc oxide, iron oxide and iron blue, and suitable organic pigments include barium, strontium, calcium and aluminum lakes and carbon black.

Pearlizing agents may be present in the composition, suitable pearlizing agents include mica coated with titanium oxide, iron oxide or natural pigments.

Fillers may be present in the composition, and suitable fillers include talc, silica, zinc stearate, mica, kaolin, nylon powder, polyethylene powder, teflon, starch, boron nitride, copolymer microspheres, such as silicone resin microspheres.

The oil phase of the compositions of the present invention may comprise one or more waxes, gums or mixtures thereof. Waxes include hydrocarbon-based waxes, fluorowaxes, and/or silicone waxes, and may be derived from vegetable, mineral, animal, and/or synthetic sources. Suitable waxes include beeswax, carnauba, candelilla, paraffin, microcrystalline, ozokerite, synthetic waxes including polyethylene waxes, silicone waxes containing C16-45. The gums are typically polydimethylsiloxanes or sodium carboxymethyl cellulose or extracts, and the semi-solid material is typically a hydrocarbon-based compound, such as lanolin and its derivatives.

The compositions of the present invention may be formulated into any suitable product form. Such product forms include, but are not limited to, aerosol sprays, creams, emulsions, solids, liquids, dispersions, foams, gels, lotions, mousses, ointments, powders, patches, pomades, solutions, hand pump sprays, sticks, masks, and wet wipes. The compositions of the present invention may be conveniently used in the preparation or as a cosmetic, dermatological or pharmaceutical topical application product by a variety of methods well known in the art.

The skin external composition of the present invention may include one or more ingredients selected from the group consisting of antiallergic agents, antimicrobial agents, antioxidants, chelating agents, colorant depigmenting agents, emollients, emulsifiers, exfoliating agents, film forming agents, fragrances, moisturizers, insect repellents, lubricants, pharmaceutically active agents, moisturizers, photostable agents, preservatives, skin care agents, skin penetration enhancers, sunscreens, stabilizers, surfactants, thickeners, viscosity modifiers, vitamins, or any combination thereof.

The invention is further illustrated below in connection with specific examples. It is to be understood that these examples are illustrative of the present invention and are not to be construed as limiting the scope of the present invention. The test methods in the following examples, in which specific conditions are not specified, are generally conducted under conventional conditions or under conditions recommended by the manufacturer. All percentages and parts are by weight unless otherwise indicated.

The experimental materials and equipment used in the following examples were as follows:

example 1 preparation of Akebia quinata extract

100G of akebia stem is weighed, extracted 2 times by 10 times of deionized water, filtered, decocted and concentrated, and a certain amount of 95% ethanol is added for ethanol precipitation (the final concentration is 70%). Filter paper filtration, concentrate the filtrate at 55 ℃, and stand at 4 ℃ overnight. Taking out, centrifuging at 3500rpm for 15min at 3 ℃, filtering with filter paper, and finally preparing into 7.31% solid solution, namely 0.731mg/mL for later use.

Example 2 preparation of Akebia quinata extract

The solid content 7.31% akebia stem extract sample prepared in example 1 was taken and added with deionized water to prepare a solid content 5% solution, i.e., 0.5mg/mL, for further use.

Example 3 preparation of Akebia quinata extract

The solid content 7.31% akebia stem extract sample prepared in example 1 was taken and added with deionized water to prepare a solid content 4% solution, i.e., 0.4mg/mL, for further use.

Example 4 preparation of Akebia quinata extract

Deionized water is added into a akebia stem extract sample with 7.31% of solid content prepared in the example 1 to prepare a solution with 1% of solid content, namely 0.1mg/mL for later use.

Example 5 preparation of Akebia quinata extract

The solid content 7.31% akebia stem extract sample prepared in example 1 is added with deionized water to prepare a solid content 0.731% solution, i.e. 0.0731mg/mL for later use.

EXAMPLE 6 evaluation of inhibition of 5-alpha reductase Activity

Testosterone is converted into Dihydrotestosterone (DHT) with stronger activity by 5 alpha-reductase in the presence of NADPH, so that the inhibition of the test substance on the 5 alpha-reductase can be measured by measuring the content change of testosterone as a substrate in a reaction system.

1. Experimental method

(1) A5-alpha reductase crude enzyme extract (RLM) is prepared by taking male SD rat, breaking head and bleeding, taking liver, placing in ice bath beaker, rinsing with ice bath physiological saline for three times, and washing off residual blood. The scissors were crushed, and 3 times of phosphate (K ₂HPO₄-KH₂PO₄, 100mM, PH7.4) -KCl (0.15M) buffer (pH 7.4) was added in a weight-volume ratio, and mechanically homogenized with a homogenizer under an ice bath. The homogenate was centrifuged at 9000 Xg for 20min at 4℃and the supernatant was taken and centrifuged at 100000 Xg for 60min at 4℃and the supernatant was discarded. Phosphate buffer solution with the same amount as the supernatant was added, and the precipitate was blown off by repeated blowing with a pipette. The mixture was centrifuged again at 100000 Xg at 4℃for 60min and the supernatant was discarded again. Adding phosphate buffer solution, and blowing and resuspending by using a suction pipe to obtain the 5-alpha reduction crude enzyme solution. Protein concentration was determined by Bardford method (using BSA as standard), diluted at 10mg/mL protein concentration and finally stored in-80℃refrigerator.

(2) The enzymatic reaction comprises the steps of adding buffer solution (Tris-HCl, pH 7.4), RLM (10 mg/mL), samples with different concentrations, dutasteride, testosterone (6 mug/mL) and NADPH (10 mM) into a 96-well plate according to the sequence, adding liquid into a reaction system, operating according to the following table, reacting at 37 ℃ for 30min, taking 50 mu L of reaction liquid after the reaction, adding 100 mu L of methanol to terminate the reaction, centrifuging 10000 mug for 10min, taking supernatant to be tested, and arranging three compound holes in each group.

TABLE 1

Note that "/" means no addition

(3) Sample preparation, preparing a solution according to the following table mode.

TABLE 2

(3) And (3) quantitative determination of testosterone, namely determining the testosterone content in the sample to be tested by adopting a UPLC-MS method, and reacting the strength of the enzyme activity by using the consumption of the substrate testosterone.

Chromatographic conditions Waters Acquity HSS T chromatographic column (100X 2.1mm,1.8 μm)

Mobile phase 0.1% formic acid water acetonitrile

Elution by gradient elution according to the following Table

TABLE 3 Table 3

Flow rate 0.4mL/min

Sample injection amount 5. Mu.L

Column temperature of 40 DEG C

The mass spectrum adopts electrospray ion source positive ion mode analysis, and the data acquisition uses a Full scan-single ion monitoring (Full scan-SIM) mode, the range is 100-1500 m/z, the resolution is 70000, the spray voltage is 3.5kV, the capillary temperature is 320 ℃, the heating temperature is 300 ℃, the sheath gas flow rate is 30Arb, the auxiliary gas flow rate is 13Arb, and the Automatic Gain Control (AGC) is 10 ⁶.

When the testosterone consumption in the sample tube was lower than that of the uninhibited group of reaction tubes, it was considered to have inhibitory 5-alpha reductase activity.

Wherein B is testosterone content of a blank control group, R is testosterone content of an inhibitor-added reaction group, and Q is testosterone content of an uninhibited reaction group.

2 Experimental results:

(1) Testosterone standard linear equation

Figure 1 shows the chromatographic results of testosterone standards.

TABLE 4 Table 4

Fig. 2 shows a linear equation for testosterone content measurement.

The standard curve is Y= 653053.4313X-2356631.8540R ² =0.9962, and testosterone concentration is in the mass concentration range of 6.25-200ng/mL, and the testosterone concentration and testosterone concentration have a better linear dose-effect relationship.

(2) 5-Alpha reductase inhibitory Activity in test samples

TABLE 5

As shown by the 5-alpha reductase inhibition experimental result, the extract of the example 1 has certain 5-alpha reductase inhibition capability when the mass concentration of the extract is 0.00731mg/mL-0.0731mg/mL as a single medicine.

The preparations of examples 1 to 5 and the like were used for the preparation of skin external preparations. The external preparation for skin is preferably a cosmetic composition such as a lotion, essence, cream, etc. The weight percentage of the preparation in the external skin preparation is 0.0005% -10% (w/w). Preferably, the weight percentage is 0.000731% -5% (w/w). More preferably, the weight percentage is 0.003655% -1% (w/w). Most preferably, the weight percentage is 0.00731% -0.5% (w/w).

The following are examples of specific applications of akebia stem extract in skin external preparations, and formulations and preparation methods of these dosage forms. In the following tables, "-" indicates no addition.

EXAMPLE 7 preparation of face cream

Example 8 preparation of emulsion

EXAMPLE 9 preparation of gel

Example 10 preparation of toning lotion

EXAMPLE 11 preparation of essence

Example 12 preparation of mask

EXAMPLE 13 preparation of eye cream

EXAMPLE 14 preparation of aerosol (cleaning foam)

EXAMPLE 15 preparation of spray

EXAMPLE 16 preparation of body washes

EXAMPLE 17 preparation of facial cleanser

Claims

1. Application of Akebia extract in skin oil control and/or hair loss prevention, wherein the Akebia extract is prepared by solvent extraction.

2. The use according to claim 1, wherein the Akebia extract is extracted using water as a solvent and is prepared by decoction extraction.

3. The use according to claim 1, wherein the Akebia extract is used at a concentration of 0.001 to 1 mg/mL.

4. The use according to claim 1, wherein the Akebia extract is used at a concentration of 0.007 mg/mL-0.5 mg/mL.

5. The use according to any one of claims 1 to 4, wherein the skin oil control and/or hair loss prevention is achieved by inhibiting 5α-reductase.

6. Use of an Akebia extract in the preparation of an external skin preparation having the effects of controlling oil content in the skin and/or preventing hair loss, wherein the Akebia extract is prepared by a solvent extraction method.

7. The use according to claim 6, wherein the Akebia extract is extracted using water as a solvent and is prepared by decoction extraction.

8. The use according to claim 6, wherein the skin oil control and/or hair loss prevention is achieved by inhibiting 5α-reductase.

9. The use according to claim 6, wherein the weight percentage of the Akebia extract in the external skin preparation is 0.0005-10 weight %, preferably 0.000731-5 weight %, more preferably 0.003655-1 weight %, and most preferably 0.00731-0.5 weight %.

10. The use according to any one of claims 6 to 9, characterized in that the skin external preparation is selected from the group consisting of: facial cream, lotion, gel, toner, essence, facial mask, eye cream, aerosol cleansing foam, spray, shower gel, and facial cleanser.