CN110731991A

CN110731991A - Skin external composition with acne removing effect

Info

Publication number: CN110731991A
Application number: CN201910694716.9A
Authority: CN
Inventors: 赵瑞丽; 王莎莎
Original assignee: Zhejiang Yangshengtang Institute of Natural Medication Co Ltd
Current assignee: Natural Medicine Institute of Zhejiang Yangshengtang Co Ltd; Zhejiang Yangshengtang Institute of Natural Medication Co Ltd
Priority date: 2019-07-30
Filing date: 2019-07-30
Publication date: 2020-01-31
Also published as: JP7422214B2; JP2022552584A; KR20220044764A; WO2021017839A1

Abstract

The present invention provides skin external compositions having an anti-acne effect, comprising (a) concentrated birch juice with a concentration of about 1.1-8 times, preferably about 1.2-4 times, more preferably about 1.2-2 times, and (B) or more selected from salicylic acid, malic acid, aloe extract, green tea extract, zinc PCA, allantoin, sodium hyaluronate and niacinamide.

Description

Skin external composition with acne removing effect

Technical Field

The present invention relates to skin external compositions having anti-acne efficacy, comprising (A) concentrated birch juice with a concentration of about 1.1-8 times, preferably about 1.2-4 times, more preferably about 1.2-2 times, and (B) or more selected from salicylic acid, malic acid, aloe extract, green tea extract, zinc PCA, allantoin, sodium hyaluronate and niacinamide.

Background

Acne, also known as whelk, pimple, comedo, pimple and the like, is kinds of chronic inflammatory skin diseases occurring in pilosebaceous units, is better to be developed in adolescents, is frequently generated in parts with vigorous sebaceous gland secretion such as faces, upper chests, backs, necks and the like, is usually characterized by multiform skin lesions such as comedo, papules, pustules, nodules and the like in clinical manifestations, and can also form scars for severe people.

It is currently widely believed that the causative factors of acne include excessive androgen levels, increased sebum secretion, hyperkeratosis of the follicular duct, and hyperproliferation of propionibacterium acnes. At present, the medicinal acne-removing products are mostly treated by using androgen antagonists, propionibacterium acnes inhibitors, tretinoin medicines and the like, although the curative effect is obvious, the toxic and side effects are large, the liver function can be damaged, and even the fetus malformation can be caused. Acne-removing cosmetics are the preferred acne prevention and treatment mode of consumers at present. The acne-removing cosmetics sold on the market at present generally have the defects of slow effect, unobvious acne-removing effect, even irritation and the like. Therefore, on the basis of the physiological and pathological characteristics of acne, the search for safe and effective acne-removing active substances is a hotspot for the development of acne-removing cosmetics and a direction of common efforts of cosmetic practitioners.

Birch juice is juice from plants of Betulaceae, and is rich in active ingredients such as polysaccharide, amino acids, vitamins, biotin, cytokinin, minerals and trace elements. The use of birch (betula) sap for face washing in europe has been documented to help improve facial acne. The birch juice from natural sources is used alone or is compounded with other active substances, and has great potential in the field of acne removal.

Disclosure of Invention

, the present invention relates to the use of a combination of (A) concentrated birch juice and (B) or more selected from salicylic acid, malic acid, aloe vera extract, green tea extract, zinc PCA, allantoin, sodium hyaluronate and niacinamide, wherein the concentrated birch juice is concentrated about 1.1-8 times, preferably about 1.2-4 times, more preferably about 1.2-2 times, in a skin external composition having anti-acne efficacy.

In another aspect, the present invention relates to skin external compositions having acne removing efficacy, comprising (a) concentrated birch juice at a concentration of about 1.1-8 times, preferably about 1.2-4 times, more preferably about 1.2-2 times, and (B) or more selected from salicylic acid, malic acid, aloe vera extract, green tea extract, zinc PCA, allantoin, sodium hyaluronate, and niacinamide.

The pathogenesis of acne involves excessive androgen levels, increased sebum secretion, follicular duct hyperkeratosis, propionibacterium acnes proliferation, and the like. Effectively reducing the vigorous sebum secretion caused by male hormone, preventing the propionibacterium acnes from hyperplasia and the excessive keratinization of the hair follicle opening and inhibiting the inflammatory reaction generated by the hyperplasia and the excessive keratinization, and is the key point for treating the acne.

Unexpectedly, the inventor finds that compared with the use of concentrated birch juice or the combination of salicylic acid, malic acid, aloe extract, green tea extract, zinc PCA, allantoin, sodium hyaluronate and nicotinamide alone, the concentrated birch juice and the combination of the salicylic acid, malic acid, aloe extract, green tea extract, zinc PCA, allantoin, sodium hyaluronate and/or nicotinamide have significantly better acne-removing efficacy which is far higher than the function superposition effect of the two, and particularly shows that the concentrated birch juice can effectively inhibit the activity of 5 α -reductase, thereby reducing the generation of dihydrotestosterone, reducing the secretion of grease, effectively inhibiting the proliferation of propionibacterium acnes, and reducing the content of IL-1 α and TNF- α in the inflammatory reaction caused by the inhibition of the activity of the 5 α -reductase, thereby showing better acne-removing efficacy.

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-acne efficacy of the concentrated birch sap is not simply linear with its concentration, but shows a tendency to increase and then decrease as the concentration factor increases, therefore, it is critical to control the concentration factor of the birch sap, which is about 1.1-8 times, preferably 1.2-4 times, more preferably 1.2-2 times in the present invention.

The (A) concentrated birch sap is contained in an amount of about 10-98% by weight, preferably about 20-98% by weight, more preferably about 30-97% by weight, based on the total weight of the composition for external use for skin having an acne removing effect.

The component (B) salicylic acid, malic acid, aloe vera extract, green tea extract, zinc PCA, allantoin, sodium hyaluronate and niacinamide are known in the art, and all of them are commercially available as they are for use in the present invention.

The total content of the component (B) is about 0.0005 to 30% by weight, preferably about 0.001 to 10% by weight, more preferably about 0.1 to 5% by weight, most preferably about 0.5 to 3% by weight, based on the total weight of the composition for external skin application having an anti-acne effect.

The skin external composition comprises a pharmaceutical composition and a cosmetic composition, especially a skin care cosmetic composition.

The composition for external application to the skin does not contain any added water, but does not exclude moisture inherently contained in the respective components.

Preferably, the composition for external application to the skin does not contain a chelating agent such as EDTA salt, sodium polyphosphate, sodium metaphosphate, gluconic acid, or the like.

The composition for external application to the skin may optionally contain, in addition to the above-mentioned components (a) and (B), components commonly used in the pharmaceutical composition or cosmetic composition of component (C), including vehicles, active ingredients, adjuvants, and the like. Component (C) is known in the art and can be selected by those skilled in the art as desired, for example, component (C) is generally present in an amount of about 0 to 70% by weight, based on the total weight of the composition.

The vehicle includes, 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 said vehicle in said cosmetic composition is known in the art, and is for example generally comprised between 0.5 and 20% of the total weight of component (C).

The active ingredients comprise antibacterial agent, antiinflammatory agent, astringent, antioxidant, humectant, emollient, oil control agent, and cutin removing component.

The antibacterial agent includes or more of, but not limited to, ursolic acid, oldenlandia diffusa flavone, honeysuckle flower extract, tea tree essential oil, chitin, cassia twig extract, zingiber corallinum volatile oil, clove extract, mushroom extract, aloe extract, artemisia leaf extract, 1-pentadecanol and derivatives thereof, cedrene, caryophyllene, longifolene, etc. the content of the antibacterial agent in the skin external composition is known in the art, and for example, it is generally 0.01-30% by weight of the total weight of component (C).

The anti-inflammatory agent includes or more of safflower yellow, dipotassium glycyrrhizinate, pollen Typhae extract, herba Paederiae extract, asiaticoside, allantoin, cucumber extract, garlic extract, burdock extract, resveratrol, radix Sophorae Flavescentis extract, cortex Magnolia officinalis extract, etc. the content of the anti-inflammatory agent in the skin external composition is known in the art, and for example, it is usually 0.01-50% by weight of the total weight of component (C).

The astringent includes or more of but not limited to green tea polyphenol, witch hazel extract, vitamin A, menthol lactate, seaweed extract, indigo naturalis, sulfur, rehmannia glutinosa, angelica, allantoin, lactic acid, tartaric acid, succinic acid, citric acid, etc. the content of the astringent in the skin external composition is known in the art, and for example, it is generally 0.01-30% by weight of the total weight of component (C).

The antioxidant includes, but is not limited to, or more of white tea polyphenols, russetose alkaloids, hawthorn flavones, coenzyme Q10, grape seed extract, asparagus extract, radish extract, L-C stock solution, safflower water extract, asiaticoside, polysaccharides from Atractylodis rhizoma, vitamin E and its derivatives, etc. the content of the antioxidant in the skin external composition is known in the art, and for example, it is generally 0.01-30% of the total weight of component (C).

Examples of the moisturizer include, but are not limited to, or more of glycerin, diglycerin, 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 glycerin, 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 rolum, pullulan, tremella polysaccharide, tamarind seed polysaccharide, and the like.

Examples of such emollients include, but are not limited to, olive oil, macadamia nut oil, sweet almond oil, grape seed oil, avocado oil, corn oil, sesame oil, soybean oil, peanut oil, meadowfoam seed oil, safflower seed oil, rosa canina oil, argan oil, jojoba seed oil, sunflower seed oil, palm kernel oil, squalane, ethylhexyl palmitate, isopropyl myristate, hydrogenated polyisobutene, isohexadecane, isododecane, diethylhexyl carbonate, dioctyl carbonate, isopropyl lauroyl sarcosinate, isononyl isononanoate, hydrogenated polydecene, tris (ethylhexanoate), cetyl ethylhexanoate, bis-diethoxydiol cyclohexane 1, 4-dicarboxylate, caprylic/capric triglyceride, oleyl erucate myristate, octyldodecanol, dimethicone, octylmethicone, cetyl dimethicone, etc. examples of solid emollients include, but are not limited to, cetyl alcohol, stearyl alcohol, cetyl stearyl alcohol, shark alcohol, behenyl wax, etc. the compositions are generally known in the amounts of these are such as the same are included in the compositions.

Examples of the oil control agent include, but are not limited to, or more of hawthorn extract, ivy extract, saxifrage extract, waxgourd seed extract, spiraea ulmaria extract, vitamin B5, verbascorbyl glycoside and puerarin, hinokitiol leaf extract, salvia miltiorrhiza root extract, green flower southern tomato bark extract, willow herb leaf extract, dendrobium extract, sage extract, avocado extract, etc. the amount of the oil control agent in the skin external composition is known in the art, and for example, it is usually 0.01 to 30% by weight based on the total weight of component (C).

The exfoliating ingredients include, but are not limited to of bromelain, salicylic acid, fruit acids, lactic acid, citric acid, enzymatic exfoliants, polyethylene beads, glycolic acid, mandelic acid, malic acid, tartaric acid, azelaic acid, acetic acid, and the like, the amount of the exfoliating ingredients in the skin topical composition is known in the art, and for example, it is typically 0.01-30% by weight of component (C).

Such adjuvants include, for example, emulsifiers, thickeners, preservatives, fragrances and the like.

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 various external emulsifiers such as are known in the art, and the content of such emulsifiers is typically 0.5% by weight of the external emulsifier.

Examples of the thickener include, but are not limited to, kinds or more of high molecular polymers such as carbomers, acrylates and derivatives thereof, xanthan gum, arabic gum, polyethylene glycol-14M, polyethylene glycol-90M, succinoglycan, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, etc. the content of the thickener in the skin external composition is known in the art, and for example, it is usually 0.1 to 10% by weight based on the total weight of the component (C).

Examples of the preservatives include, but are not limited to, kinds 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, and the content of the preservatives in the skin external composition is known in the art, and for example, it is usually 0.01 to 50% by weight based on the total weight of the component (C).

The skin external composition of the present invention may 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 commonly used in the cosmetic field. 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 pharmaceutical composition or cosmetic composition in various dosage forms such as liquid, lotion, cream or gel can be prepared according to the need, wherein the cosmetic composition can be in various forms such as lotion, spray, emulsion, essence lotion or essence emulsion, BB cream, sunscreen cream, etc.

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 different concentrations of birch juice, salicylic acid, malic acid, and combinations thereof on 5 α -reductase activity 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 2 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 2mLEP tube, adding a proper amount of crude enzyme extract according to the proportion of 1:4, breaking the mixture into homogenate at 4 ℃ by a wall breaking machine, freezing and centrifuging at 4 ℃ at a high speed (10000 rpm/min, 10 min), and taking supernatant fluid for storage 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) blank control determination, namely respectively taking 2 groups of 200 mu LEP tubes (4 in each group), adding 5 α -reductase crude enzyme, PBS solution (pH is 7.4) and reduced coenzyme II into the 2 groups of 200 mu LEP tubes, mixing the two groups of , immediately putting the two groups of tubes into boiling water for boiling for 5 minutes, centrifuging the two groups of tubes, absorbing 50 mu L of liquid for subsequent Elisa detection to obtain the initial testosterone content of the blank control group, mixing the other groups of tubes, putting the mixture into a constant-temperature shaking table for uniformly mixing for 60 minutes, putting the mixture into boiling water for boiling for 5 minutes, centrifuging the mixture, absorbing 50 mu L of liquid for subsequent Elisa detection, and taking the difference value of the initial content and the initial content as the converted testosterone content of;

(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％＝(ΔA₀-ΔA_n)/ΔA₀x100％

wherein:

ΔA₀control testosterone reduction;

ΔA_ntestosterone reduction in the inhibitor group.

The test results are shown in the following table.

Sample (I)	Inhibition rate
		Stock solution of birch juice	19％±2％
2 times concentrated birch juice	48％±4％
		Concentrated birch juice 10 times	26％±3％
Salicylic acid	10％±1％
		Malic acid	9％±1％
Birch juice stock solution and salicylic acid	35％±6％
		Birch juice stock solution and malic acid	38％±5％
2 times of concentrated birch juice and salicylic acid	68％±9％
		2 times of concentrated birch juice and malic acid	77％±7％
10 times concentrated birch juice and salicylic acid	32％±3％
		10 times concentrated birch juice + malic acid	34％±5％

The results show that the effect of the combination of the 2-time concentrated birch juice and the salicylic acid or the malic acid is obviously better than that of the single birch juice stock solution, the 2-time concentrated birch juice and the salicylic acid or the malic acid in the aspect of inhibiting the 5 α -reductase, and simultaneously is also obviously better than that of the combination of the birch juice stock solution and the salicylic acid or the malic acid.

Example 2: experiment for inhibiting bacteria

According to the evaluation method of the antibacterial and bacteriostatic effects of QB/T2738-.

1. Experimental Material

Equipment and consumables: filter paper (bacteriostatic carrier); a vernier caliper.

Reagent: nutrient agar culture medium; and (4) bacterial suspension.

The strain source is as follows: propionibacterium acnes was purchased from the China center for type culture Collection.

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.

2. Testing

The operation steps of the bacteriostasis experiment are as follows:

(1) preparing the bacteriostatic tablets: taking a circular sterile dried filter paper sheet with the diameter of 5mm, dropwise adding 20 mu L of a liquid sample in the center of the bacteriostatic sheet, then flatly placing the filter paper sheet in a clean sterile plate, uncovering the filter paper sheet, and placing the filter paper sheet in a constant temperature box (37 ℃) for drying, or placing the filter paper sheet at room temperature for natural drying for later use.

(2) Inoculation of test bacteria: dipping with sterile cotton swab to obtain a concentration of 1 × 10⁴cfu/ml～9×10⁴The cfu/ml test bacterial suspension is evenly smeared on the surface of a nutrient agar medium plate for 3 times, the plate is rotated for 60 degrees every time the smearing is carried out for 1 time, and finally, a cotton swab is smeared around the edge of the plate for weeks, the plate is covered, and the plate is placed at room temperature for drying for 5 minutes.

(3) Sticking bacteriostatic test samples: and (3) placing 1 infectious bacterium flat plate on each test, and placing 4 test sample plates and 1 reference sample plate on each flat plate for 5 plates in total. And (3) removing the sample pieces by using sterile forceps, and pasting the sample pieces on the surface of the flat plate, wherein the distance between the centers of the sample pieces is more than 25mm, and the distance between the centers of the sample pieces and the periphery of the flat plate is more than 15 mm. After the sample is well stuck, the sample is lightly pressed by using sterile forceps to be tightly stuck to the surface of the flat plate, the flat plate is covered, the flat plate is placed in a constant temperature box at 37 ℃, and the result is observed after the culture is carried out for 16 to 18 hours. The diameter of the antibacterial ring (including the patch) was measured with a vernier caliper and recorded. The experiment was repeated 3 times.

The test results are shown in the following table.

The results of the bacteriostatic test show that compared with the single use of the birch juice stock solution, the 2-time concentrated birch juice and the PCA zinc and the combination of the birch juice stock solution and the PCA zinc, the combination of the 2-time concentrated birch juice and the PCA zinc obviously improves the capacity of inhibiting the growth of propionibacterium acnes. When the birch juice is concentrated to 10 times, the effect of combining with other active matters is obviously weakened, even the effect of combining the birch juice stock solution with other active matter raw materials is not as good as the effect of combining with other active matter raw materials.

Example 3: rat auricle model experiment

In this example, based on murine auricle model experiments, the ability of various concentrations of birch juice and aloe vera extract, green tea extract, allantoin, niacinamide, and combinations thereof against IL-1 α, TNF- α in serum were examined, respectively.

1. Material

Experimental animals: SPF grade SD male healthy rats, body mass (200. + -. 20) g, were provided from Hospital, Zhejiang province.

Drugs and reagents are 100% oleic acid, ELISA IL-1 α kit and ELISA TNF- α kit.

2. Method of producing a composite material

(1) The modeling method randomly divided the rats into 2 groups, wherein 8 rats in the normal group were used, and the rest were used as modeling groups. 100% oleic acid was applied to the right auricle skin of each rat in the model group by a 1ml syringe, 1 time per day, about 0.3ml each time, and continuously applied for 3 weeks. Coating with 100% oleic acid to day 22, 2 molding rats were randomly selected, 2 ears were cut from the right ear where oleic acid was coated, fixed with 10% formaldehyde, embedded in paraffin, sectioned, HE stained, and histopathological changes were observed under light microscopy. The observation under the light lens can be seen: the cuticle is thickened and hyperkeratosis appears, the hair follicle is dilated, the hair follicle opening and the infundibulum are filled with keratotic substances, the keratotic plug is blocked, the opening is enlarged to form a pot shape, and the sebaceous gland disappears. According to the histological judgment grading standard of experimental acnes, the experimental acnes model of the auricle of the rat is prompted to be formed.

(2) After animal grouping and drug administration molding are successful, the rest rats in the molding group are randomly divided into model blank groups and birch juice groups with different concentrations according to the body constitution, and each group comprises 8 rats. On day 22 of the experiment, topical application of the spread was started. The dose of each group is 0.1ml, the dose position is the rat right auricle modeling position, 1 time/day, and 2 weeks of continuous administration. The normal group and the model group were not coated with any drug.

(3) Collecting and detecting method of specimen, taking femoral artery blood of each group of animals 24 hours after last administration, separating serum, storing at-20 ℃ for later use, and measuring the content of IL-1 α and TNF- α in serum by adopting a double antibody sandwich ABC-ELISA method.

(4) 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.

Note: indicates significance compared to the model group, p-value less than 0.05; indicates extreme significance compared to the model, with p values less than 0.01.

The above results show that the combined use of the birch juice concentrate 2 times with the aloe extract, the green tea extract, the allantoin or the nicotinamide significantly improves the ability of IL-1 α, TNF- α in antiserum compared to the use of the birch juice concentrate 2 times with the birch juice concentrate, the birch juice concentrate 2 times with the green tea extract, the allantoin or the nicotinamide alone, and the combination of the birch juice concentrate with the aloe extract, the green tea extract, the allantoin or the nicotinamide.

Example 4: oil content of human skin

In this example, the effect of different concentrations of birch juice and zinc PCA, sodium hyaluronate, and their compositions on the oil content of human skin, respectively, was examined.

, the skin oil secretion is excessive and is the main reason of acne, the experiment uses a skin oil determinator (German CK company SM815) to carry out quantitative measurement on the skin oil gland secretion before and after the cosmetics are used, the anti-acne efficacy of the cosmetics can be evaluated, the SM815 determination principle is based on the photometer principle, special extinction adhesive tapes with the thickness of 0.1mm absorb the oil on the skin of a human body and become semitransparent adhesive tapes, the light transmission quantity of the adhesive tapes changes, the more the absorbed oil is, the larger the light transmission quantity is, and the content of the skin oil can be measured.

The experimental process comprises selecting experimental patients (20-35 years old, half each for male and female) with excessive oil secretion, grouping the experimental patients into 8 groups, cleaning face on the same day of oil testing, and smearing 3mg/cm on cheek 30 min later²Different products of (2). Skin oil content was dynamically measured at 0 hour, 2 hours, 4 hours, and 8 hours using SM815, respectively, and skin oil content change was calculated.

The experimental evaluation and the test of the testers are carried out in a constant temperature and humidity laboratory by a double-blind marking method, the temperature is 25 +/-1 ℃, and the humidity is 60% +/-5%.

The test results are shown in the following table.

The content of the skin oil in the control group gradually increases along with the time, which indicates that the testee has vigorous oil secretion. The sample set results show that the use of 2-fold concentrated birch juice in combination with zinc PCA or sodium hyaluronate inhibited the hypersecretion of oil in the subjects more significantly than the use of birch juice stock alone, 2-fold concentrated birch juice, zinc PCA, sodium hyaluronate, and combinations of birch juice stock and zinc PCA or sodium hyaluronate. When the birch juice is concentrated to 10 times, the effect of combining with other active matters is obviously weakened, even the effect of combining the birch juice stock solution with other active matter raw materials is not as good as the effect of combining with other active matter raw materials.

Example 5: preparation and performance investigation of acne-removing essence milk

The formula of the acne-removing essence containing different active substances is shown in the following table:

the preparation process comprises the following steps: weighing the phase A raw material, uniformly mixing all components in the phase A, ensuring that the components are completely dissolved, heating to 60 ℃, and preserving heat for 20 minutes. And (3) cooling to 45 ℃ while stirring, sequentially adding the phase B, and uniformly stirring to obtain the multiple acne-removing essence milk products.

In order to test the actual using effect of the product, the different acne removing essence milk products are respectively subjected to crowd trial investigation, samples are distributed to patients with acne for trial, the age is 20-35 years, each product subject is 60 persons, trial lasts for 1 month, meanwhile, questionnaires are filled, and finally, the results are counted.

The criteria for ranking were as follows:

has remarkable effect that the healing or the skin lesion is reduced by more than 60 percent

Effective, the skin damage is reduced by 20 to 60 percent

Ineffective, regression of skin lesions < 20% or aggravated.

The results of the trial survey of the population are as follows.

Sample (I)	Significant effect/example	Effective/example	Invalid/example
				Blank sample	0	3	57
Stock solution of birch juice	24	14	22
				2 times concentrated birch juice	30	12	18
Salicylic acid	11	14	35
				Birch juice stock solution and salicylic acid	31	15	14
2 times of birch juice stock solution and salicylic acid	38	14	8

The above results indicate that the essence milk containing the combination of the 2-fold concentrated birch sap and the salicylic acid has more remarkable acne removing capability compared with the acne removing essence milk containing the birch sap stock solution, the 2-fold concentrated birch sap, the salicylic acid and the combination of the birch sap stock solution and the salicylic acid which are used alone.

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) or more selected from salicylic acid, malic acid, aloe vera extract, green tea extract, zinc PCA, allantoin, sodium hyaluronate and niacinamide, wherein the concentrated birch juice is concentrated 1.1-8 times, preferably 1.2-4 times, more preferably 1.2-2 times, in a composition for external use on skin having anti-acne efficacy.
2. The use of claim 1, wherein the composition for external application to the skin does not comprise any added water.
3. The use according to claim 1 or 2, wherein the composition for external application to the skin is a pharmaceutical composition or a cosmetic composition.
skin external composition having acne removing effect, comprising (A) concentrated birch juice with concentration of 1.1-8 times, preferably 1.2-4 times, more preferably 1.2-2 times, and (B) or more substances selected from salicylic acid, malic acid, aloe extract, green tea extract, zinc PCA, allantoin, sodium hyaluronate and nicotinamide.
5. The skin external composition according to claim 4, wherein the content of the concentrated birch sap of the component (A) is 10 to 98% by weight, preferably 20 to 98% by weight, more preferably 30 to 97% by weight, based on the total weight of the skin external composition.
6. The composition for external skin application according to claim 4 or 5, wherein the content of the component (B) is 0.0005 to 30% by weight, preferably 0.001 to 10% by weight, more preferably 0.1 to 5% by weight, most preferably 0.5 to 3% by weight, based on the total weight of the composition for external skin application.
7. The topical skin composition of any one of claims 4-6, , wherein the topical skin composition does not include any added water.
8. The topical skin composition of any one of claims 4-7, , wherein the topical skin composition is a pharmaceutical composition or a cosmetic composition.