WO2021017839A1 - External skin composition having anti-acne effect - Google Patents

Abstract

Disclosed is an external skin composition having an anti-acne effect, comprising (A) a concentrated birch sap, the concentration factor thereof being about 1.1 - 8 times, preferably about 1.2 - 4 times, and more preferably about 1.2 - 2 times, and (B) one or more materials selected from salicylic acid, malic acid, an aloe extract, a green tea extract, PCA zinc, allantoin, sodium hyaluronate and nicotinamide.

Description

External skin composition with anti-acne effect Technical field

The present invention relates to an external skin composition with anti-acne effect, which comprises (A) concentrated birch tree juice with a concentration ratio of about 1.1-8 times, preferably about 1.2-4 times, more preferably about 1.2-2 times , And (B) one or more substances selected from salicylic acid, malic acid, aloe extract, green tea extract, PCA zinc, allantoin, sodium hyaluronate and nicotinamide.

Background technique

Acne, also known as acne, acne, acne, acne, etc., is a chronic inflammatory skin disease that occurs in the sebaceous gland unit of the hair follicle. It is more common in adolescents, and often occurs in the sebaceous glands of the face, upper chest, back, and neck. In vigorous areas, the clinical manifestations are often characterized by pleomorphic skin lesions such as acne, papules, pustules, and nodules. In severe cases, scars may also form. Acne has a high incidence, often lasts for a long time, and is easy to recur. For some acne, if treatment is not taken in time, it will cause hyperpigmentation and even scarring, which will seriously affect the patient's appearance and bring serious negative effects on the patient's psychology.

At present, it is generally believed that the pathogenic factors of acne include excessive androgen levels, increased sebum secretion, hyperkeratosis of the hair follicle duct, and excessive proliferation of Propionibacterium acnes. At present, medicinal anti-acne products mostly use androgen antagonists, P. acnes inhibitors, and tretinoin drugs. Although the effect is obvious, the toxic and side effects are also relatively large, which may cause liver damage or even cause infant deformity. Acne-repellent cosmetics are currently the first choice for acne prevention and treatment by consumers. However, acne-removing cosmetics currently on the market generally suffer from deficiencies such as slow effectiveness, insignificant acne-removing effects, and even irritation. Therefore, finding safe and effective anti-acne active substances based on the physiological and pathological characteristics of acne is a hot spot in the development of anti-acne cosmetics, and it is also the direction of joint efforts of cosmetics practitioners.

Birch sap is the sap from the birch family plant, rich in active ingredients such as polysaccharides, amino acids, vitamins, biotin, cytokinins, minerals and trace elements needed by the human body. The use of birch (European white birch) sap to wash your face in Europe has been reported to help improve facial acne. Naturally sourced birch sap used alone or in combination with other active substances has great potential in the field of anti-acne.

Summary of the invention

In one aspect, the present invention relates to (A) concentrated birch tree juice and (B) selected from salicylic acid, malic acid, aloe extract, green tea extract, PCA zinc, allantoin, sodium hyaluronate and nicotinamide The use of one or more of the combination in the skin external composition with anti-acne effect, wherein the concentration of the concentrated birch tree juice is about 1.1-8 times, preferably about 1.2-4 times, more preferably about 1.2-2 times.

In another aspect, the present invention relates to a skin external composition with anti-acne effect, which comprises (A) concentrated birch tree juice, the concentration of which is about 1.1-8 times, preferably about 1.2-4 times, more preferably about 1.2-2 times, and (B) one or more substances selected from salicylic acid, malic acid, aloe extract, green tea extract, PCA zinc, allantoin, sodium hyaluronate, and nicotinamide.

The pathogenesis of acne involves excessive androgen levels, increased sebum secretion, hyperkeratosis of the hair follicle duct, and proliferation of Propionibacterium acnes. Effectively reduce the exuberant sebum secretion caused by male hormones, prevent the proliferation of P. acnes and hyperkeratosis of the hair follicle mouth, and inhibit the resulting inflammation, which is the key to the treatment of acne.

Unexpectedly, the inventors found that compared with the use of concentrated birch tree juice or salicylic acid, malic acid, aloe extract, green tea extract, PCA zinc, allantoin, sodium hyaluronate, and nicotinamide alone, The combination of concentrated birch juice with salicylic acid, malic acid, aloe extract, green tea extract, PCA zinc, allantoin, sodium hyaluronate and/or nicotinamide has significantly better anti-acne efficacy, much higher than The superimposing effect of the two functions is specifically manifested in the ability to effectively inhibit the activity of 5α-reductase, thereby reducing the production of dihydrotestosterone, reducing the secretion of oil, and effectively inhibiting the proliferation of Propionibacterium acnes and reducing the inflammation caused by it. In the content of IL-1α and TNF-α, it shows better anti-acne effect.

The birch sap involved in the present invention is obtained from the genus Betula, Betula alba, Betula pubescens, Betula pendula and Asian white birch (Betula platyphylla). Varieties. The birch sap is a colorless, transparent, no-sediment-free, and nutrient-rich sap that is artificially collected by drilling holes in the base of the birch trunk between thawing and early spring. The birch sap is commercially available and used as it is, for example, it can be purchased from Daxinganling Chaoyue Wild Berry Development Co., Ltd.

The concentrated birch sap in the present invention is obtained by concentrating the above-mentioned commercial products. Concentration methods are known in the art, such as heating concentration, low-temperature vacuum concentration, membrane concentration and the like. In the present invention, the concentration is preferably carried out by a low-temperature freeze concentration or membrane concentration process. For example, the commercially available birch juice stock solution is input into a low-temperature drying equipment, the temperature is lowered to -40°C to -70°C, and the vacuum is applied to 0.1-30Pa. Concentrated in vacuum at low temperature to obtain concentrated birch sap with different concentration times.

Furthermore, the inventors also found that the acne-removing effect of concentrated birch sap and its concentration degree is not a simple linear relationship, but as the concentration ratio increases, it first increases and then decreases. Therefore, controlling the concentration ratio of birch sap is critical. In the present invention, controlling the concentration ratio of birch sap is about 1.1-8 times, preferably 1.2-4 times, more preferably 1.2-2 times.

The content of the (A) concentrated birch sap is about 10-98% by weight, preferably about 20-98% by weight, more preferably about 30-97% by weight, based on the total amount of the skin external composition with anti-acne effect weight.

The component (B) salicylic acid, malic acid, aloe extract, green tea extract, PCA zinc, allantoin, sodium hyaluronate and nicotinamide are known in the art, and they are all commercially available, Used in the present invention as it is.

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

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

The external skin composition does not contain any added water, but does not exclude the moisture inherently contained in each component.

Preferably, the external skin composition does not contain chelating agents such as EDTA salt, sodium polyphosphate, sodium metaphosphate, and gluconic acid.

In addition to the aforementioned components (A) and (B), the skin external composition may optionally include component (C) commonly used ingredients in pharmaceutical compositions or cosmetic compositions, including vehicles, active ingredients and Accessories etc. Component (C) is known in the art, and those skilled in the art can select its type and amount according to needs. For example, the content of component (C) is usually about 0-70% by weight, based on the total of the composition. weight.

The vehicle includes, for example, a diluent, a dispersant, or a carrier, and examples thereof include, but are not limited to, ethanol, dipropylene glycol, butylene glycol, and the like. The content of the vehicle in the cosmetic composition is known in the art, for example, it usually accounts for 0.5-20% of the total weight of component (C).

The active ingredients include antibacterial agents, anti-inflammatory agents, astringents, antioxidants, moisturizers, emollients, oil control agents, exfoliating ingredients and the like.

The antibacterial agents include, but are not limited to, ursolic acid, oldenlandia diffusa flavonoids, honeysuckle flower extract, tea tree essential oil, chitin, cinnamon twig extract, coral ginger volatile oil, clove extract, mushroom extract, aloe extract, One or more of mugwort leaf extract, 1-pentadecanol and its derivatives, cedarene, caryophyllene, and phyllene. The content of the antibacterial agent in the skin external composition is known in the art, for example, it usually accounts for 0.01-30% of the total weight of the component (C).

The anti-inflammatory agent includes, but is not limited to, safflower yellow, dipotassium glycyrrhizinate, cattail pollen extract, arrowroot extract, asiaticoside, allantoin, cucumber extract, garlic extract, burdock extract, white One or more of Veratrol, Sophora flavescens extract, Magnolia Bark extract, etc. The content of the anti-inflammatory agent in the skin external composition is known in the art, for example, it usually accounts for 0.01-50% of the total weight of the component (C).

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

The antioxidants include, but are not limited to, white tea polyphenols, red moss alkaloids, hawthorn flavonoids, coenzyme Q10, grape seed extract, asparagus extract, radish extract, L-C stock solution, safflower water extract, Centella asiatica One or more of glycosides, Atractylodes polysaccharides, vitamin E and its derivatives. The content of the antioxidant in the skin external composition is known in the art, for example, it usually accounts for 0.01-30% of the total weight of the component (C).

Examples of the moisturizer include, but are not limited to, glycerin, diglycerin, butylene glycol, propylene glycol, 1,3-propanediol, dipropylene glycol, 1,2-pentanediol, polyethylene glycol-8, polyethylene glycol Alcohol-32, methylglucitol-10, methylglucitol-20, PEG/PPG-17/6 copolymer, glycerol-7, glycerol-26, glycerol glucoside, PPG-10 methyl glucose ether, PPG-20 methyl glucose ether, PEG/PPG/polybutylene 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, Sclerotium hydrolysate One or more of gum, short-stemming enzyme polysaccharide, tremella polysaccharide, caper seed polysaccharide, etc. The content of the moisturizer in the skin external composition is known in the art, for example, it usually accounts for 1-30% of the total weight of the component (C).

Examples of the emollient 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, white flower seed oil, safflower seed oil, Dogtooth rose hip oil, argan tree kernel oil, jojoba seed oil, sunflower seed oil, palm tree fruit oil, squalane, ethylhexyl palmitate, isopropyl myristate, hydrogenated polyisobutylene, isotene Hexane, isododecane, diethylhexyl carbonate, dioctyl carbonate, isopropyl lauroyl sarcosine, isononyl isononanoate, hydrogenated polydecene, glycerol tri(ethylhexanoate) , Cetyl alcohol ethyl hexanoate, bis-diethoxy diethylene glycol cyclohexane 1,4-dicarboxylate, caprylic/capric triglyceride, oleyl erucate, octyldodecanol Myristate, Octyldodecanol, Dimethicone, Octyl Dimethicone, Cetyl Dimethicone, Cyclopenta Dimethicone, etc. One or more. Examples of solid emollients include, but are not limited to, cetyl alcohol, stearyl alcohol, cetearyl alcohol, behenyl alcohol, scylitol, lauric acid, myristic acid, palmitic acid, stearic acid, beeswax, candelilla Tree wax, carnauba wax, lanolin, ozokerite, jojoba seed wax, paraffin wax, microcrystalline wax, hydrogenated rice bran wax, hydrogenated coconut oil glycerides, glyceryl behenate/eicosanate, myristyl alcohol One or more of myristate, bis-diglyceride polyacyl adipate-2, shea butter, and muluxing palm seed butter. The content of the emollient in the skin external composition is known in the art, for example, it usually accounts for 1-50% of the total weight of component (C).

Examples of the oil control agent include, but are not limited to, hawthorn extract, ivy extract, saxifrage extract, winter melon seed extract, spirea extract, vitamin B5, verbascum glycosides and puerarin, Japanese cypress leaf extract One or more of extracts, Salvia miltiorrhiza root extract, Ennan tomato bark extract, Willow orchid leaf extract, Dendrobium extract, Sage extract, Avocado extract, etc. The content of the oil control agent in the external skin composition is known in the art, for example, it usually accounts for 0.01-30% of the total weight of the component (C).

The exfoliating ingredients include but are not limited to bromelain, salicylic acid, fruit acid, lactic acid, citric acid, enzyme cutin exfoliant, polyethylene beads, glycolic acid, mandelic acid, malic acid, tartaric acid, azelaic acid, acetic acid, etc. One or more of. The content of the exfoliating ingredient in the skin external composition is known in the art, for example, it usually accounts for 0.01-30% of the total weight of the component (C).

The auxiliary materials include, for example, emulsifiers, thickeners, preservatives, perfumes and the like.

Examples of the emulsifier include, but are not limited to, cetearyl oleate, sorbitan oleate, polysorbate-60, polysorbate-80, methylglucose sesquistearic acid Ester, PEG-20 methyl glucose sesquistearate, PEG-40 hydrogenated castor oil, PPG-26-butanol-26, PEG-4 polyglycerol-2 stearate, PEG-60 hydrogenated Castor oil, steareth-2, steareth-21, PPG-13-decyltetradeceth-24, cetearyl glucoside, PEG-100 stearate, glycerin Stearate, Glyceryl Stearate SE, Coco Glucoside, Ceteareth-25, PEG-40 Stearate, Polyglyceryl-3 Methyl Glucose Distearate, Glyceryl stearate citrate, polyglyceryl-10 stearate, polyglyceryl-10 myristate, polyglyceryl-10 dioleate, polyglyceryl-10 laurate, polyglyceryl-10 isostearate Fatty acid ester, polyglyceryl-10 oleate, polyglyceryl-10 diisostearate, polyglyceryl-6 laurate, polyglyceryl-6 myristate, sucrose stearate, sucrose polystearate One or more of acid esters. The content of the emulsifier in the skin external composition is known in the art, for example, it usually accounts for 0.5-10% of the total weight of the component (C).

Examples of the thickener include, but are not limited to, carbomers, acrylic acid (ester) and its derivatives, xanthan gum, gum arabic, polyethylene glycol-14M, polyethylene glycol-90M, succinyl poly One or more of high molecular polymers such as sugar, hydroxyethyl cellulose, hydroxypropyl cellulose, and hydroxypropyl methyl cellulose. The content of the thickener in the external skin composition is known in the art, for example, it usually accounts for 0.1-10% of the total weight of the component (C).

Examples of the preservatives include, but are not limited to, methyl paraben, ethyl paraben, propyl paraben, phenoxyethanol, benzyl alcohol, phenethyl alcohol, bis(hydroxymethyl)imidazolidinyl urea, potassium sorbate, One or more of sodium benzoate, chlorphenesin, sodium dehydroacetate, etc. The content of the preservative in the skin external composition is known in the art, for example, it usually accounts for 0.01-50% of the total weight of the component (C).

The skin external composition of the present invention can be prepared by any suitable method known in the art. For example, it can be prepared using equipment such as dissolving tanks, emulsifying pots, dispersers, and delivery pumps commonly used in the cosmetics field. When preparing, put the water-soluble substance into the water-phase dissolving kettle, and the oil-soluble substance into the oil-phase dissolving kettle, and heat the temperature of the two kettles to about 80°C. For the raw materials that are easy to agglomerate, they can be mixed with a disperser first. Pre-dispersion. After the dissolution is completed, the oil phase and the water phase are transported to the emulsifying pot for homogenization and emulsification for about 5-15 minutes. After the emulsification is completed, the temperature of the material body is reduced to normal temperature, optional flavors, preservatives, etc. are added, and the pH of the product is adjusted as necessary. The products can be filled and shipped only after the relevant test indicators are qualified.

The above preparation methods can be deleted or adjusted according to dosage form requirements. Pharmaceutical compositions or cosmetic compositions in various dosage forms such as liquids, emulsions, creams, creams or gels can be prepared according to needs, wherein the cosmetic compositions can be lotions, sprays, emulsions, essence water or essence milk, BB cream , Sunscreen and other forms.

Example

Hereinafter, the present invention will be further described in detail with reference to the embodiments. However, it should be understood that these examples and comparative examples are only used to describe the present invention in more detail, and should not be understood as limiting the scope of the appended claims of the present invention in any form.

Example 1 : Inhibition of 5α-reductase activity

In this example, the effects of birch sap, salicylic acid, malic acid, and their combination on the activity of 5α-reductase of different concentration multiples were investigated and compared.

1. Concentration of birch sap

The fresh white birch sap stock solution purchased from Daxinganling Chaoyue Wild Berry Development Co., Ltd. is fed into the low-temperature drying equipment, the temperature is reduced to -65°C, the vacuum is reduced to 0.1Pa, and the concentration is 2 and 10 times.

2. Test

Experimental instruments: balance, wall breaker, constant temperature oscillator, high-speed refrigerated centrifuge, microplate reader.

Experimental reagents and consumables:

Finasteride: Shanghai Modern Pharmaceutical Co., Ltd. (tablet 5mg/tablet);

Reduced coenzyme Ⅱ (NADPH): American Roche company (powder 10mg);

Testosterone Elisa kit: Wuhan Ulsheng Company (Cat. No.: CEA458Ge);

5α-reductase crude enzyme: self-provided;

PBS: Wuhan Boster Company;

Phenylmethylsulfonyl fluoride (PMSF): American Sigma company;

Dithiothreitol (DTT): American Sigma company;

1M Tris-HCl: Wuhan Boster Company.

Sample adding information:

The sample amount of a single raw material is one whole raw material; the compound raw material is half a birch sap raw material plus half a 0.1% other active raw material.

In vitro 5α-reductase activity influence analysis steps are as follows:

(1) Preparation of crude enzyme: Take 5 normal healthy mice, let their necks dislocated to death, open the abdominal cavity, take their testes, divide them into several portions, place them in a 2mL EP tube, add an appropriate amount of crude enzyme extract in a ratio of 1:4, The homogenate was crushed by a wall breaker at 4°C and centrifuged at a high speed at 4°C (10000rpm/min, 10 minutes), and the supernatant was stored at 4°C. BCA method is used to determine protein concentration, and follow-up experiments can be carried out after 1 mg/mL;

(2) Determination of blank control: Take 2 groups of 200μLEP tubes (4 in each group), add 5α-reductase crude enzyme, PBS solution (pH=7.4), reduced coenzyme II, and put them in immediately after mixing. Boil in boiling water for 5 minutes. After centrifugation, 50μL of liquid is drawn for subsequent Elisa detection, which is the initial testosterone content of the blank control group; the other group is mixed and placed in a constant temperature shaker for 60 minutes, put in boiling water and boiled for 5 minutes, after centrifugation Aspirate 50μL of liquid for subsequent Elisa detection, and the difference from the initial content is the testosterone content of the blank control group after transformation;

(3) Determination of sample group: Take 2 groups of 200μL EP tubes (4 in each group), add 5α-reductase crude enzyme, PBS solution (pH=7.4), reduced coenzyme II and test materials, and mix in one group Immediately put it in boiling water and boil for 5 minutes. After centrifugation, 50μL of liquid was drawn for subsequent Elisa detection, which is the initial testosterone content of the inhibitor group; the other group was mixed and placed in a constant temperature shaker to mix for 60 minutes, and then boiled in boiling water for 5 After centrifugation, 50 μL of liquid was drawn for subsequent Elisa detection. The difference between the initial content and the testosterone content of the inhibitor group was the testosterone content after conversion. Each experiment has 4 parallel samples.

The inhibition rate of 5α-reductase is calculated according to the following formula:

I％＝(△A ₀ -△A _n )/△A ₀ x100％

among them:

△A ₀ ---- Decrease of testosterone in the control group;

△A _n ---- Decrease of testosterone in inhibitor group.

The test results are shown in the table below.

sample	Inhibition rate
Birch sap	19%±2%
2 times concentrated birch sap	48%±4%
10 times concentrated birch sap	26%±3%
Salicylic acid	10%±1%
Malic acid	9%±1%
Birch sap extract + salicylic acid	35%±6%
Birch sap extract + malic acid	38%±5%
2 times concentrated birch sap + salicylic acid	68%±9%
2 times concentrated birch sap + malic acid	77%±7%
10 times concentrated birch sap + salicylic acid	32%±3%
10 times concentrated birch juice + malic acid	34%±5%

The above results show that in terms of inhibiting 5α-reductase, the combination of 2 times concentrated birch sap and salicylic acid or malic acid is significantly better than using birch sap stock solution, 2 times concentrated birch sap and salicylic acid alone. Or malic acid, which is also significantly better than the combination of birch sap and salicylic acid or malic acid. When the birch sap is concentrated to 10 times, the effect of using it in combination with other active substances is significantly weakened, and is even inferior to the combined effect of birch sap stock and other active ingredients.

Example 2 : Antibacterial Experiment

According to the QB/T 2738-2012 method for evaluating the antibacterial and antibacterial effects of daily chemical products, the birch sap, PCA zinc, and their compositions of different concentrations were tested by the antibacterial ring method.

1. Experimental materials

Equipment and consumables: filter paper (bacteriostatic agent carrier); vernier calipers.

Reagents: nutrient agar medium; bacterial suspension.

Source of strain: Propionibacterium acnes was purchased from the China Type Culture Collection.

Sample addition information: the sample amount of a single raw material is a whole raw material; the compound raw material is half a birch sap raw material plus half a 0.1% other active raw material.

2. Test

Operation steps of antibacterial experiment:

(1) Preparation of antibacterial tablets: take a circular sterile dry filter paper with a diameter of 5mm, drop 20μL of liquid sample into the center of the antibacterial tablet, and then place the filter paper flat in a clean sterile plate with the lid open Dry in a constant temperature oven (37°C), or dry naturally at room temperature for later use.

(2) Inoculation of test bacteria: dip a sterile cotton swab into a test bacteria suspension with a concentration of 1×10 ⁴ cfu/ml to 9×10 ⁴ cfu/ml, and smear it evenly on the surface of the nutrient agar medium plate 3 times. After each application, the plate should be rotated 60 degrees, and finally use a cotton swab to apply a circle around the edge of the plate. Cover the plate and let it dry at room temperature for 5 minutes.

(3) Placement of antibacterial test samples: Place 1 stained plate for each test, 4 pieces of test specimens for each plate, 1 piece of control specimens, a total of 5 pieces. Use sterile tweezers to remove the samples and place them on the surface of the plate. The distance between the centers of the samples is more than 25mm and the distance between the centers of the samples is more than 15mm. After the paste is placed, use sterile tweezers to gently press the sample piece to make it close to the surface of the plate, cover the plate, place it in a 37°C incubator, and observe the results after incubating for 16 to 18 hours. Measure the diameter of the antibacterial ring (including the patch) with a vernier caliper and record it. The test was repeated 3 times.

The test results are shown in the table below.

sample	Diameter of inhibition zone of Propionibacterium acnes/mm
blank	0
Birch sap	12±2
2 times concentrated birch sap	25±4
10 times concentrated birch sap	15±3
PCA zinc	8±1
Birch sap extract + PCA zinc	20±3
2 times concentrated birch sap + PCA zinc	38±5
10 times concentrated birch sap + PCA zinc	18±5

Bacteriostatic test results showed that the combined use of birch sap concentrate and PCA zinc was significantly greater than the combination of birch sap concentrate, birch sap concentrate, PCA zinc, and the combination of birch sap concentrate and PCA zinc alone. Improved the ability to inhibit the growth of Propionibacterium acnes. When the birch sap is concentrated to 10 times, the effect of using it in combination with other active substances is significantly weakened, and is even inferior to the combined effect of birch sap stock and other active ingredients.

Example 3 : Rat ear pinna model experiment

In this example, based on the mouse ear model experiment, different concentrations of birch sap and aloe extract, green tea extract, allantoin, nicotinamide, and their combinations were tested against serum IL-1α and TNF. -Alpha ability.

1. Material

Experimental animals: SPF-grade SD male healthy rats, weighing (200±20) g, provided by Zhejiang Medical Academy.

Drugs and reagents: 100% oleic acid; ELISA IL-1α kit; ELISA TNF-α kit.

Sample addition information: the sample amount of a single raw material is a whole raw material; the compound raw material is half a birch sap raw material plus half a 0.1% other active raw material.

2. Method

(1) Modeling method The rats were randomly divided into 2 groups, of which 8 were in the normal group and the rest were modeled. Apply 100% oleic acid into the skin of each rat's right auricle with a 1ml syringe, once a day, about 0.3ml each time for 3 weeks. Apply 100% oleic acid to the 22nd day, randomly select 2 model rats, cut off the 2 ears of the right ear where the oleic acid is applied, fix with 10% formaldehyde, embed in paraffin, section, HE stain, light Observe histopathological changes under the microscope. Observation under the light microscope shows that the stratum corneum is thickened, hyperkeratosis, the hair follicle is expanded, the mouth and funnel of the hair follicle are filled with keratinized material, the horn plug is blocked, and the expansion is pot-shaped, and the sebaceous gland disappears. According to the grading standard of experimental acne histology, it is suggested that the experimental acne model of rat auricle has been formed.

(2) Animal grouping and drug administration After successful modeling, the remaining rats in the model were randomly divided into a blank model group and a birch sap group with different concentrations, with 8 rats in each group. On the 22nd day of the experiment, topical smearing was started. The dosage of each group was smeared 0.1ml, and the administration site was the model site of the right auricle of the rat, once a day, for 2 weeks. No drugs were applied to the normal group and model group.

(3) Collection and detection methods of specimens 24 hours after the last administration, the femoral artery blood of each group of animals was taken, serum was separated, and stored at -20°C for later use. The serum IL-1α and TNF-α levels were determined by the double antibody sandwich ABC-ELISA method.

(4) GraphPad Prism Program software was used for drawing, and T-test statistical analysis was used between groups.

The test results are shown in the table below.

sample	Only count	IL-1α(ng/ml)	TNF-α(ng/ml)
normal group	8	14.36±0.39	6.31±1.52
Model group	8	22.56±1.32	22.47±0.65
Birch sap	8	19.83±1.03*	18.50±1.34*
2 times concentrated birch sap	8	17.65±0.88**	14.34±0.86**
10 times concentrated birch sap	8	19.99±1.22*	18.48±1.80*
Aloe extract	8	20.36±1.03*	17.93±1.48*
Green tea extract	8	19.22±1.03*	19.05±1.93*

Allantoin	8	19.01±2.16*	18.47±1.65*
Nicotinamide	8	20.68±2.02*	19.54±1.05*
Birch sap extract + aloe extract	8	16.98±1.54**	13.81±1.22**
Birch tree juice extract + green tea extract	8	17.32±0.99**	15.46±0.98**
Birch sap extract + allantoin	8	16.33±1.87**	12.55±0.74**
Birch sap extract + niacinamide	8	17.64±1.22**	14.74±0.55**
2 times birch sap extract + aloe extract	8	14.87±0.82**	8.72±0.52**
2 times birch tree juice extract + green tea extract	8	14.37±1.31**	6.69±1.35**
2 times birch sap extract + allantoin	8	14.85±1.49**	7.49±1.51**
2 times birch sap extract + niacinamide	8	15.62±1.01**	6.85±1.44**
10 times birch sap extract + aloe extract	8	17.45±1.65**	15.22±1.53**
10 times birch tree juice extract + green tea extract	8	18.97±0.87**	14.83±0.56**
10 times birch sap extract + allantoin	8	18.21±1.54**	16.53±0.98**
10 times birch sap extract + niacinamide	8	17.43±1.31**	15.38±1.35**

Note: * indicates that it is significant compared with the model group, and the p value is less than 0.05; ** indicates that it is extremely significant compared to the model, and the p value is less than 0.01.

The above results show that, compared with the use of birch tree juice stock, 2 times concentrated birch tree juice, aloe extract, green tea extract, allantoin or nicotinamide, as well as birch juice stock and aloe extract, green tea extract, allanto Compared with the combination of nicotin or nicotinamide, the combination of 2 times concentrated birch tree juice and aloe extract, green tea extract, allantoin or nicotinamide significantly improved the ability of antiserum IL-1α and TNF-α. When the birch sap is concentrated to 10 times, the effect of using it in combination with other active substances is significantly weakened, and is even inferior to the combined effect of birch sap stock and other active ingredients.

Example 4 : Human skin oil content

In this example, the effects of different concentrations of birch tree sap, PCA zinc, sodium hyaluronate, and their composition on the oil content of human skin were investigated.

Excessive skin oil secretion is one of the main causes of acne. In the experiment, a skin oil tester (SM815, a German CK company) was used to quantitatively measure the skin oil gland secretions before and after using cosmetics, which can evaluate the anti-acne efficacy of cosmetics. The SM815 measurement principle is based on the photometer principle. After a special matt tape with a thickness of 0.1mm absorbs the oil on the human skin, it will become a translucent tape, and its light transmission will change and the absorbed grease The more, the greater the light transmission, so that the content of skin oil can be measured.

Sample addition information: the sample amount of a single raw material is a whole raw material; the compound raw material is half a birch sap raw material plus half a 0.1% other active raw material.

Experimental process: select patients (20 to 35 years old, half male and female) for hypersecretion of oil, the testers are divided into groups, 8 in each group, clean the face on the day of the oil test, and rub 3mg/cm ² on the cheek after 30 minutes. product. The skin oil content was dynamically measured with SM815 at 0 hour, 2 hours, 4 hours, and 8 hours, respectively, and the skin oil content changes were calculated.

Both the experiment evaluation and the testers were double-blind labeling method, and the test was carried out in a constant temperature and humidity laboratory with a temperature of 25±1°C and a humidity of 60%±5%.

The test results are shown in the table below.

The skin oil content of the control group gradually increased over time, indicating that the tester had strong oil secretion. The results of the sample group showed that compared with the birch sap stock solution alone, the birch sap of 2 times concentrated birch sap, PCA zinc, sodium hyaluronate, and the combination of birch sap stock and PCA zinc or sodium hyaluronate, the birch sap was 2 times concentrated. The combined use of tree sap and PCA zinc or sodium hyaluronate more significantly inhibited the tester’s excessive oil secretion. When the birch sap is concentrated to 10 times, the effect of using it in combination with other active substances is significantly weakened, and is even inferior to the combined effect of birch sap stock and other active ingredients.

Example 5 : Preparation and performance investigation of anti-acne essence milk

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

The preparation process is as follows: Weigh the raw materials of phase A, mix the components in phase A uniformly and ensure that the components are completely dissolved, heat to 60°C and keep it for 20 minutes. When the temperature is lowered to 45°C while stirring, add phase B one by one and stir evenly to obtain the above-mentioned various acne-removing essence milk products.

In order to test the actual use effect of the product, the above-mentioned different acne essence milk products were tested separately, and samples were distributed to patients with acne, aged 20-35 years old, 60 people for each product, one trial Month, fill out the questionnaire at the same time, and finally count the results.

The classification criteria are as follows:

Significantly effective: recovery or skin lesions subsided>60%

Effective: 20%～60% of skin lesions subsided

Invalid: skin lesions subsided <20% or worsened.

The results of the population trial survey are as follows.

sample	Significantly effective/case	Effective/case	Invalid/case
Blank sample	0	3	57
Birch sap	twenty four	14	twenty two
2 times concentrated birch sap	30	12	18
Salicylic acid	11	14	35
Birch sap extract + salicylic acid	31	15	14
2 times birch sap extract + salicylic acid	38	14	8

The above results show that compared with the single use of birch tree juice concentrate, birch tree juice concentrate, salicylic acid, and a combination of birch tree juice concentrate and salicylic acid, it contains two times more concentrated birch juice Essence milk combined with salicylic acid has a more pronounced anti-acne ability.

The technical solutions of the above-mentioned embodiments are preferred implementations of the present invention. Several improvements and changes can be made without departing from the principle of the present invention, and these improvements and changes should also be regarded as falling within the protection scope of the present invention.

Claims (8)

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