WO2021017846A1 - Enhanced anti-aging cosmetic composition - Google Patents

Abstract

An enhanced anti-aging cosmetic composition, comprising (A) a concentrated birch sap, the concentration factor thereof being about 1.05 - 8 times, preferably 1.1 - 4 times, and more preferably 1.2 - 2 times, and (B) one or more materials selected from a polypeptide, sodium hyaluronate, retinol, retinene, retinol palmitate, tocopherol, ubiquinone and adenosine.

Description

Enhanced anti-aging cosmetic composition Technical field

The present invention relates to an enhanced anti-aging cosmetic composition comprising (A) concentrated birch tree sap, the concentration of which is 1.05-8 times, preferably 1.1-4 times, more preferably 1.2-2 times, (B) From one or more of polypeptides, sodium hyaluronate, retinol, retinal, retinol palmitate, tocopherol, ubiquinone and adenosine.

Background technique

Birch is a deciduous tree of the birch family. There are currently about 100 species in the world, mainly distributed in the northern temperate zone and the cold temperate zone. Among them, there are about 29 varieties in my country, mainly distributed in the northeast, northwest, north and southwest. Birch trees are mostly grown in remote mountainous areas with little human intervention and no industrial pollution. Birch sap (also called birch sap) is the fresh sap from the birch bark being cut or the trunk is drilled. It is colorless or light yellow, free of precipitation and impurities, and has a light birch fragrance. The birch sap contains a lot of sugars, amino acids, vitamins, biotin, cytokinins, trace mineral elements, aromatic oils, betulin, saponin and other compounds, which have good skin care and anti-aging effects.

Skin aging means that the skin's ability to protect and adjust to the body is reduced, and it cannot adapt to changes in internal and external environments, and changes in overall appearance such as color and shape. It is often manifested as reduced or disappeared skin elasticity, loose skin, increased wrinkles, and dry skin , Thinning, easy desquamation, itching, dull complexion, dull, and accompanied by stains and warts, affecting appearance and confidence. With the increasingly serious problem of population aging and the continuous improvement of living standards, consumers' anti-wrinkle awareness continues to increase, and more and more consumers are eager to use cosmetics to maintain their youthful appearance. According to statistics, anti-wrinkle products have become the category with the fastest growth rate and greatest potential in the Chinese cosmetics market.

Summary of the invention

In one aspect, the present invention relates to (A) concentrated birch sap and (B) selected from polypeptides, sodium hyaluronate, retinol, retinal, retinol palmitate, tocopherol, ubiquinone and adenosine The use of one or more of the combination in an enhanced anti-aging cosmetic composition, wherein the concentration factor of the concentrated birch sap is 1.05-8 times, preferably 1.1-4 times, more preferably 1.2-2 times.

In another aspect, the present invention provides an enhanced anti-aging cosmetic composition comprising (A) concentrated birch sap with a concentration ratio of 1.05-8 times, preferably 1.1-4 times, more preferably 1.2-2 times, And (B) one or more substances selected from polypeptide, sodium hyaluronate, retinol, retinal, retinol palmitate, tocopherol, ubiquinone and adenosine.

The anti-aging cosmetic composition of the present invention can promote the synthesis of collagen I, III, IV, VII, XVII, elastin, laminin, etc. in the extracellular matrix, reduce the expression of metal matrix protease MMP-1, and promote hyalurin Acid production strengthens the structural integrity and functionality of the epidermis, dermis, and dermal-epidermal junctions, improves skin elasticity, reduces wrinkles, and at the same time has the effects of improving the skin barrier, reducing transdermal water loss, anti-inflammatory and soothing, thereby providing significant anti-inflammatory effects. Ageing, especially anti-wrinkle formation effect.

The anti-aging cosmetic composition does not contain any added water, but does not exclude the moisture inherently contained in each component.

In a preferred embodiment, chelating agents such as EDTA salt, sodium polyphosphate, sodium metaphosphate, and gluconic acid are not added to the anti-aging cosmetic composition.

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 no-drug sap that is artificially collected by drilling holes at the base of the trunk of the birch from thawing to 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 used 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. Concentrate in vacuum at low temperature to obtain concentrated birch sap with a concentration ratio of 1.05-8 times.

Unexpectedly, the present inventors found that compared with the non-concentrated birch sap stock, the concentration ratio is 1.05-8 times, preferably 1.1-4 times, and more preferably 1.2-2 times concentrated birch sap and polypeptide, The combination of sodium hyaluronate, retinol, retinal, retinol palmitate, tocopherol, ubiquinone, and/or adenosine can significantly provide the anti-aging effect of the cosmetic composition, especially anti-wrinkle formation. This indicates that the combination of the concentrated birch sap and the polypeptide, sodium hyaluronate, retinol, retinal, retinol palmitate, tocopherol, ubiquinone and/or adenosine produced synergy or enhancement. Effect.

The content of the (A) concentrated birch sap is about 18-98% by weight, preferably about 20-95% by weight, more preferably about 22-92% by weight, most preferably about 30-90% by weight, based on the anti-aging The total weight of the cosmetic composition.

The polypeptide in the component (B) is known in the art to promote the synthesis of matrix proteins, especially collagen, and may also increase the production of elastin, hyaluronic acid, glycosaminoglycans and fibronectin. Those that provide anti-wrinkle effects. Examples of polypeptides that can be used in the present invention include but are not limited to dipeptide-2, dipeptide-4, tripeptide-1, tetrapeptide-21, pentapeptide-3, hexapeptide-3, hexapeptide-8, Peptide-9, hexapeptide-10, palmitoyl dipeptide-5, palmitoyl dipeptide-7, palmitoyl tripeptide-1, palmitoyl tripeptide-5, palmitoyl tetrapeptide-3, palmitoyl tetrapeptide-7 , Palmitoyl pentapeptide-3, palmitoyl pentapeptide-4, palmitoyl hexapeptide-6, palmitoyl oligopeptide, acetyl tetrapeptide-3, acetyl tetrapeptide-5, acetyl tetrapeptide-9, acetyl Pentapeptide-3, acetylhexapeptide-3, acetylhexapeptide-8, acetyloctapeptide-1, carnosine, copper peptide, oligopeptide-1, myristoyl pentapeptide-11, dipeptide diaminobutane Acyl benzyl amide diacetate, decarboxyl carnosine HCL, and any combination thereof, preferably palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, acetyl hexapeptide-3, dipeptide diaminobutyryl benzyl amide two Acetate, palmitoyl oligopeptide, hexapeptide-9 and any combination thereof. The polypeptide is preferably selected from palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, acetylhexapeptide-3, dipeptide diaminobutyrylbenzylamide diacetate, palmitoyl oligopeptide, hexapeptide- 9 and any combination thereof. The above substances are all commercially available.

Examples of the sodium hyaluronate in the component (B) include, but are not limited to, large molecular weight sodium hyaluronate (1800-2200KDa), medium molecular weight sodium hyaluronate (1000-1800KDa), small molecular weight sodium hyaluronate ( One or more of 10-1000KDa), hydrolyzed sodium hyaluronate (1-10KDa), acetylated sodium hyaluronate, etc. Sodium hyaluronate can be prepared by methods known in the art, for example, firstly, sodium hyaluronate is prepared from a plant-derived medium (peptone, glucose, etc.) by fermentation. Sodium hyaluronate is also commercially available, for example, from Huaxi Freda Biomedical Co., Ltd.

The retinol, retinal, retinol palmitate, tocopherol, ubiquinone, and adenosine in the component (B) are also ingredients known in the art, all of which are commercially available.

The content of the component (B) is about 0.005-10% by weight, preferably about 0.01-8% by weight, more preferably about 0.1-6% by weight, most preferably about 0.5-5% by weight, based on the anti-aging cosmetic composition The total weight of the object.

In addition to the aforementioned components (A) and (B), the anti-aging cosmetic composition may optionally include components (C) commonly used in skin care cosmetics, examples of which include, but are not limited to, vehicles, active ingredients, and Accessories etc. These ingredients are known in the art, and those skilled in the art can select the type and amount of component (C) according to their needs. For example, the content of component (C) is usually about 0-70% by weight, based on the anti- The total weight of the aging cosmetic composition.

The vehicle is those known in the art, such as diluents, dispersants and carriers, 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 are those known in the art, and examples thereof include, but are not limited to, for example, emollients, moisturizers, other anti-wrinkle active ingredients, and the like.

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 cosmetic composition is known in the art, for example, it usually accounts for 1-50% of the total weight of 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 cosmetic 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 other anti-wrinkle active ingredients include, but are not limited to, hydrolyzed collagen, hydrolyzed elastin, yeast extract, oryzanol, tetrahydrocurcumin, ellagic acid, whey protein, salicyl sphingosine, silymarin, silk gum Protein, tocopherol phosphate sodium, allantoin, ribonucleic acid (RNA), grape (VITIS VINIFERA) seed extract, palm tree (PTEROCARPUS) MARSUPIUM bark extract, tea (CAMELLIA SINENSIS) polyphenols, wine extract , Apple seed extract, FAGUS SYLVATICA bud extract, ADANSONIA DIGITATA extract, Artemia (ARTEMIA) extract, IRIS FLORENTINA root extract, Hesperidin , Ginsenosides, salvia miltiorrhiza (SALVIA MILTIORRHIZA) extract, nicotinamide, ursolic acid, lycopene, coffee (COFFEA ARABICA) extract, lactic acid, superoxide dismutase (SOD), evening primrose (OENOTHERA BIENNIS) oil , Ceramide, dipalmitoyl hydroxyproline, hydroxystearic acid, salicylic acid, ergothioine, lysolecithin, lipoic acid, glycogen, resveratrol, ferulic acid, soluble yeast One or more of cytoplasm, lactic acid bacteria fermentation lysate, etc. The content of the other anti-wrinkle active ingredients in the cosmetic composition is known in the art, for example, it usually accounts for 0.01-10% of the total weight of component (C).

Examples of the auxiliary materials include, but are not limited to, 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 cosmetic 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 cosmetic 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, methylparaben, propylparaben, phenoxyethanol, benzyl alcohol, phenethyl alcohol, bis(hydroxymethyl)imidazolidinylurea, potassium sorbate, sodium benzoate, chlorobenzene Glycerol, sodium dehydroacetate, caprylic hydroxamic acid, 1,2-hexanediol, 1,2-pentanediol, p-hydroxyacetophenone, caprylyl glycol, glyceryl caprylate, undecylenic acid One or more of glycerides, sorbitan caprylate, ethylhexylglycerol, tree peony root extract, etc. The content of the preservative in the cosmetic composition is known in the art, for example, it usually accounts for 0.01-2% of the total weight of component (C).

The anti-aging cosmetic composition of the present invention can be prepared by any suitable method known in the art. For example, it can be prepared by 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. According to needs, the anti-aging cosmetic composition can be made into various dosage forms such as creams, creams, lotions, and essences.

Example

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

Example 1 : Preparation of composition A, A1, B, B1, B2, B3, B4, C, C1, C2, D, D1, D2, E, E1, E2, F, F1, F2, J

The specific formulations of the above-mentioned compositions are as follows:

Note: The data in the above table represents the weight% of each component, based on the total weight of the composition, the same below.

The above compositions are prepared as follows:

1. Input the fresh white birch sap raw liquid purchased from Daxinganling Chaoyue Wild Berry Development Co., Ltd. into the low-temperature drying equipment, cool to -65℃, vacuum to 0.1Pa, and concentrate to 1.2 times, 4 times and 9 times;

2. Mix raw materials 1, 2, 3, 4, 5, 6, 7, 8, 9, and 11 to obtain the corresponding composition.

Example 2 : Compositions A, A1, B, B1, B2, B3, B4, C, C1, C2, D, D1, D2, E, E1, E2, F, F1, F2, J are associated with aging Effects of gene expression

In this example, the above compositions A, A1, B, B1, B2, B3, B4, C, C1, C2, D, D1, D2, E, E1, E2, F, F1, F2 were tested and compared The effect of J1 on the expression of aging-related genes.

Experimental instruments: fluorescence quantitative PCR instrument (Roche), ultra-clean workbench (Su Jing), carbon dioxide incubator (Binder), microplate reader (BIO-TEK), micro oscillator.

Experimental reagents and consumables: human primary fibroblasts, 6-well plates, fibroblast culture medium, RNA extraction kit, reverse transcription kit, Trizol lysis solution, etc.

The steps of gene expression analysis based on fibroblasts are as follows:

(1) Inoculation: Inoculate cells into a 6-well plate at a seeding density of 5E5/well, and incubate overnight in a 37°C and 5% CO ₂ incubator;

(2) Administration: When the plating rate of cells in the 6-well plate reaches about 60%, add the test substance of each composition, and set 6 multiple wells for each composition;

(3) Sample collection: After incubating at 37°C and 5% CO _{2 for} 24 hours, discard the culture solution, add 1 mL Trizol to each well, pipette to lyse the cells, and collect the sample;

(4) PCR detection: RNA is extracted, reverse transcribed to cDNA, and then subjected to fluorescence quantitative PCR detection;

(5) Analysis: The 2- ^△△CT method is used for calculation of the results, and the T-Test method is used for statistical analysis.

The test results are shown in the table below.

sample	Collagen I	Collagen III	Collagen IV	Collagen VII	Elastin	Laminin
A	2.95±0.09	1.29±0.16	1.19±0.12	1.18±0.02	4.28±0.27	0.89±0.05
A1	9.95±0.37	3.52±0.05	2.29±0.34	1.69±0.07	8.84±0.52	1.54±0.13
B	4.52±0.38	1.59±0.19	1.87±0.19	1.67±0.24	5.58±0.51	1.56±0.17
B1	11.23±0.83	4.68±0.23	4.79±0.22	4.77±0.19	15.13±1.17	4.36±0.22
B2	19.01±1.66	6.97±0.49	7.63±0.37	6.39±0.34	20.27±2.15	5.21±0.29
B3	18.78±1.62	6.65±0.51	7.04±0.60	5.96±0.47	19.08±1.53	5.17±0.44
B4	9.69±1.21	4.18±0.32	3.62±0.47	3.25±0.36	11.54±1.01	3.83±0.22
C	3.02±0.26	1.18±0.07	1.24±0.11	1.35±0.09	4.47±0.38	0.96±0.04
C1	9.83±1.05	4.10±0.33	4.28±0.42	4.56±0.37	13.06±1.22	3.19±0.33
C2	18.18±1.76	6.85±0.53	6.81±0.57	6.10±0.45	19.21±1.88	5.36±0.19
D	4.28±0.37	1.94±0.09	1.75±0.13	1.71±0.18	5.39±0.45	1.52±0.11
D1	10.97±1.22	4.86±0.31	4.64±0.29	4.61±0.43	14.87±1.66	4.12±0.36
D2	18.53±1.39	7.03±0.50	7.21±0.65	6.42±0.51	19.88±1.49	5.45±0.37
E	3.62±0.33	1.17±0.09	1.44±0.12	1.27±0.13	4.55±0.43	1.02±0.08
E1	10.12±1.08	4.23±0.34	4.38±0.31	4.42±0.40	13.29±1.18	3.77±0.35
E2	18.72±1.67	7.15±0.63	7.04±0.55	6.17±0.62	18.98±1.69	5.53±0.46
F	3.84±0.23	1.31±0.10	1.26±0.15	1.49±0.17	4.37±0.34	1.10±0.09
F1	10.67±1.24	4.16±0.35	4.36±0.43	4.46±0.39	13.11±1.27	3.94±0.41
F2	19.37±0.68	6.92±0.73	6.84±0.63	6.12±0.57	19.19±2.36	5.83±0.38
J	1.08±0.12	1.02±0.06	1.01±0.02	1.04±0.08	1.10±0.01	1.00±0.09

The above results show that compared with the blank control J, the other compositions can increase the expression levels of genes such as collagen I, III, IV, VII, elastin, and laminin, indicating that they contain birch sap, concentrated birch sap, palm The combination of acyl tripeptide-5, sodium hyaluronate, retinol, tocopherol, and/or ubiquinone all have a certain anti-wrinkle effect. However, comparing the results of composition B2, C2, D2, E2, F2 and composition A1, B, C, D, E, and F respectively, it can be clearly seen that the 1.2-fold concentrated birch sap and palmitoyl tripeptide-5 The combination of sodium hyaluronate, retinol, tocopherol, or ubiquinone significantly increased the gene expression level of each protein, which fully demonstrated the synergy between them; in particular, the comparison The results of composition B2, B3, B4, C2, D2, E2, F2 and composition B1, C1, D1, E1, and F1 can clearly be seen that compared with the non-concentrated birch sap, the concentrated birch sap The combination with polypeptide, sodium hyaluronate, retinol, tocopherol or ubiquinone significantly improves the anti-wrinkle effect of the cosmetic composition, and the concentration factor of the concentrated birch sap is about 1.05-8 times, preferably about 1.1-4 times .

Example 3 : The influence of compositions G, G1, G2 on the expression of aging-related proteins

In this example, the effects of the above-mentioned compositions G, G1, and G2 on the expression of aging-related proteins were further tested and compared.

Experimental equipment: ultra-clean workbench (Su Jing), plate washer (BIO-RAD), microplate reader (BIO-TEK), carbon dioxide incubator (Binder)

Experimental reagents and consumables: human primary fibroblasts, 12-well plates, fibroblast culture medium, ELISA test kits with different indicators, etc.

experimental method:

(1) Inoculation: Inoculate in a 12-well culture plate at an inoculation density of 2E5/well, culture at 37°C and 5% CO _{2 in an} incubator, and change the medium every two days;

(2) Dosing: When the cell fusion reaches more than 60% again, add different groups of test substances, each group has 6 multiple holes;

(3) Receiving samples: After 48 hours in a 37°C and 5% CO ₂ incubator, discard the culture solution, add 1 mL Trizol to each well, pipette to lyse the cells, and collect the samples;

(4) Detection: Perform index determination according to the determination method of ELISA kit;

(5) Analysis: T-Test method is used for statistical analysis.

The test results are shown in the table below.

The above results indicate that, compared with adenosine alone, the compositions G1 and G2 containing birch sap or 1.2 times concentrated birch sap and adenosine significantly increased the protein expression levels of aging-related proteins in keratinocytes , Indicating that birch sap or concentrated birch sap has a synergistic effect with adenosine. Furthermore, comparing G1 and G2, it can be seen that the synergistic effect of concentrated birch sap on peptides is significantly higher than that of the original birch sap.

Example 4 : Preparation of compositions H, H1, H2 and J1

H: Palmitoyl Tripeptide-5

H1: Birch sap stock solution + palmitoyl tripeptide-5

H2: 1.2 times concentrated birch sap + palmitoyl tripeptide-5

J1: Blank control (water-based, no birch sap, no peptides)

The formula of each of the above compositions is as follows:

The above compositions are prepared as follows:

1. Oil phase: add 6, 8, 9, 10, 11, 13, 14, 15, 16 raw materials into the oil phase pot, and heat each raw material to 80°C, dissolve and mix well;

2. Mix No. 4, 17, and 19 raw materials evenly at room temperature;

3. Mix the 12, 18, 21 raw materials and other raw materials uniformly at room temperature;

4. Aqueous phase: Heat the raw materials 1, 2, 3, 5, and 7 to 80°C, add the mixture of step 2, dissolve, and mix well;

5. Emulsification: add the water phase and the oil phase to the emulsification tank, keep the temperature at 80°C, homogenize and emulsify at 3000rpm for 5 minutes, add No. 20 raw material after the emulsification is completed;

6. Stir and add the mixture of No. 22 and step 3 when the temperature is lowered to 40°C, mix well and discharge to obtain each composition.

Example 5 : Effect of composition H, H1, H2, J1 on photoaging 3D full-thickness skin model

In this example, the effects of the above-mentioned cream compositions H, H1, H2, and J1 on the photo-aging 3D full-thickness skin model were tested and compared.

Experimental equipment: ultra-clean workbench (Su Jing), carbon dioxide incubator (Binder), UVA and UVB irradiator, microplate reader (BIO-TEK), cryostat (Leica).

Experimental reagents and consumables: 3D full-thickness skin model (made in the laboratory), ELISA test kits with different indicators, MTT kits, etc.

The experimental steps are as follows:

(1) 3D full-thickness model construction: use keratinocytes and fibroblasts to construct a 3D skin model;

(2) Modeling and drug administration: When the model leaves the factory, that is, day 0, SSUV irradiation treatment (UVA: 30J/cm2; UVB: 50mJ/cm2) is carried out every day, and then a certain amount of samples are respectively coated on the corresponding model surface , The model control is only coated with the model culture medium, each of the composition has 6 multiple holes, smeared once a day, and the total duration of action is 4 days;

(3) Sampling and testing: After the completion of the sample, collect the culture medium and use the ELISA kit for MMP1 determination. In addition, take 3 models from each group and use the MTT kit for the determination of model viability. Staining to determine the content of collagen IV at the junction of the true epidermis;

(4) Analysis: Use Test method for statistical analysis.

The test results are shown in the table below.

sample	Organizational vitality	Collagen IV expression at the junction of true epidermis	MMP1(ng)
Model comparison	1.355±0.099	0.218±0.045	9.083±0.881
H	2.091±0.212	1.778±0.165	4.323±0.411
H1	3.996±0.333	4.745±0.213	2.467±0.411
H2	4.252±0.441	5.565±0.488	2.007±0.215
J1	1.383±0.116	0.288±0.076	8.916±0.774

The above results show that, compared with the composition H, the compositions H1 and H2 can improve the tissue viability of the 3D full-thickness skin model, promote the expression of collagen IV at the junction of the true epidermis, and significantly reduce the expression of MMP-1 in the extracellular matrix. This fully shows that the combination of birch sap or concentrated birch sap and polypeptide can significantly improve the anti-wrinkle effect, indicating that the two have a significant synergistic effect. Furthermore, comparing H1 and H2, it can be seen that the synergistic effect of the concentrated birch sap on the polypeptide is significantly higher than that of the original birch sap.

Example 6 : Effect of composition H and H2 on human skin morphology

In this example, subjects with wrinkle grades 1-5 were used as the research objects. The test volunteers used H and H2 for 2 weeks and 4 weeks on the left and right side of the eye wrinkles, cheek moisture and TEWL changes, and evaluated the combination The effects of H and H2 on anti-wrinkle, increase skin moisture content and improve skin barrier function.

Using the half-face control test method, the volunteers were tested as follows before using the product, 2 weeks and 4 weeks after using the product:

1) Use primos to take pictures of the outer corners of the left and right sides of the volunteers' eyes, and use the software to calculate parameters, including the number of wrinkles, wrinkle area, wrinkle depth, etc.;

2) Use Corneometer to test the moisture content of the cheek skin on the left and right sides;

3) Test the transcutaneous water loss rate (TEWL value) of the left and right cheeks with Tewameter.

After cleansing in the morning and evening, after using toner, take appropriate test samples and apply evenly on the left and right faces (including around the eyes). The original skin care habits remain unchanged throughout the test process, and only test samples are used to replace the previous creams used by the subjects. .

The test equipment used is:

Primos (Canfield, USA)

Corneometer (Courage & Khazaka, Germany)

Tewameter (Courage & Khazaka, Germany)

Subjects: Subjects who meet the following conditions will be selected:

1) According to the wrinkle grade standard photos, screen out subjects with wrinkle grade 1-5, and wrinkles not caused by disease;

2) The tested part has not received skin treatment, beauty and other tests that may affect the results;

3) Those who have not used hormone drugs or immunosuppressive agents in the past month;

4) Those who have not participated in other clinical trials at present or in the last three months.

Among them, those who meet any of the following will be excluded:

1) Pregnant or lactating women;

2) Those with severe systemic diseases, immunodeficiency or autoimmune diseases;

3) Those who have used antihistamines in the past week or have used immunosuppressants in the past month;

4) Those who have used any anti-inflammatory drugs at the test site in the past two months;

5) Insulin dependent diabetic patients;

6) Patients with respiratory diseases being treated;

7) People with allergies, allergic dermatitis, etc., and those with a history of skin diseases or diseases;

8) People who are receiving dermatological treatment;

9) The test area has large areas of birthmarks, scratches, leukoplakia, pigmented moles, keloids and other skin signs that affect the test.

other requirements:

1) During the test, it is forbidden to use other products that affect the test;

2) During the test, use the same facial fixation facility and fixation method;

3) Test environment, test time and test location:

The test process is as follows:

1) The test areas are the outer corners of the eyes, forehead and cheeks on both sides. Before the test, the subjects cleaned the face with facial cleanser, and wiped it with a dust-free absorbent paper towel after cleaning;

2) Sit quietly for 20 minutes in a room that meets the standard, without drinking water or beverages, and the test area is exposed, stay relaxed, and avoid touching the test area;

3) The left and right corners of the eyes are randomly divided into product application area and blank control area to ensure a statistical balance;

4) During the test, use the same facial fixation facility and fixation method. According to the primos instructions, take pictures of wrinkles on the left and right outer corners of the eyes of the subjects before use, 2 weeks and 4 weeks after use, and use Corneometer and Tewameter to test both sides of the volunteers. Cheek moisture value and TEWL value.

Number of participants: 39 were enrolled, 5 dropped out halfway, and a total of 34 effective volunteers, with an average age of 45.5±8.5 years.

The test results are as follows.

(1) Number of wrinkles

Table 1: Test results of the number of wrinkles (mean±SD value, n=34)

Number of wrinkles	H2	H
Before using the test product (W0)	137.69±53.09	131.31±51.77
After using the test product for 2 weeks (W2)	83.14±48.41**ΔΔ	126.69±49.32
After using the test product for 4 weeks (W4)	75.72±39.6**ΔΔ	123.59±45.86

Note: "**" means p<0.01 compared with the product before use, "ΔΔ" means p<0.01 compared with H.

The number of wrinkles is used to characterize the number of wrinkles in the corners of the eyes. The higher the value, the more wrinkles, and vice versa. The number of skin wrinkles gradually decreased after H2 was used, and the number of wrinkles decreased significantly at 4 weeks (p<0.01). There was no significant difference in wrinkle area after H using for 4 weeks compared with before using (P>0.05). After 4 weeks of use, the difference between H2 and H is very significant (p<0.01), indicating that H2 has the effect of reducing skin wrinkles.

(2) Wrinkle depth

Table 2: Wrinkle depth test results (mean ± SD value, n=34)

Wrinkle depth (um)	H2	H
Before using the test product	79.59±35.2	76.52±36.3
After using the test product for 2 weeks	75.72±45.3	75.33±43.9
After using the test product for 4 weeks	69.31±32.4**Δ	73.03±39.4

Note: "**" means p<0.01 compared with the product before use, and "Δ" means p<0.01 compared with H.

Wrinkle depth is used to characterize the depth of wrinkles at the corners of the eyes. The higher the value, the deeper the wrinkles, and vice versa. The depth of skin wrinkles gradually decreased after H2 was used, and the wrinkle depth decreased significantly at 4 weeks (p<0.01). There was no significant difference in wrinkle depth after H using for 4 weeks compared with before using (P>0.05). After 4 weeks of use, the difference between H2 and H was significant (p<0.05), indicating that H2 has the effect of reducing skin wrinkles.

(3) Wrinkle area

Table 3: Test results of wrinkle area (mean±SD value, n=34)

Wrinkle area	H2	H
Before using the test product	80.74±18.02	79.56±19.34
After using the test product for 2 weeks	67.3±18.81**Δ	75.1±19.57
After using the test product for 4 weeks	62.5±22.09**Δ	73.75±20.93

Note: "**" means p<0.01 compared with before using the product; "Δ" means p<0.05 compared with H.

Wrinkle area is used to represent the total area of wrinkles in the corners of the eyes. The higher the value, the more wrinkles, and vice versa. The skin wrinkle area gradually decreased after H2 use, and the wrinkle area decreased significantly at 4 weeks (p<0.01). There was no significant difference in wrinkle area after H using for 4 weeks compared with before using (P>0.05). After 4 weeks of use, the difference between H2 and H is significant (p<0.01), indicating that H2 has a significant effect of reducing skin wrinkles.

(4) Moisture content

Table 4: Test results of moisture content (average value ± SD value, n=34)

Moisture	H2	H
Before using the test product	33.14±11.36	32.18±10.97
After using the test product for 2 weeks	38.94±12.25*Δ	33.29±11.54
After using the test product for 4 weeks	42.18±10.98**ΔΔ	34.62±11.66

Note: "*" means p<0.05 compared with the product before use, "**" means p<0.01 compared with the product before use; "Δ" means p<0.05 compared with H, "ΔΔ" means p<0.05 compared with H Ratio p<0.01.

Moisture content is used to characterize the moisture content of the stratum corneum of the skin. The higher the value, the higher the skin moisture content, and vice versa, the lower the skin moisture content. The skin moisture content gradually increased after H2 use, and the increase was very significant at 4 weeks (p<0.01). H after 4 weeks of use, the skin moisture content was not significantly different from that before use (P>0.05). After 4 weeks of use, the difference between H2 and H is very significant (p<0.01), indicating that H2 has the effect of increasing skin moisture content.

(5)TEWL

Table 5: TEWL test results (mean±SD value, n=34)

TEWL	H2	H
Before using the test product	19.7±8.01	18.9±8.23
After using the test product for 2 weeks	16.0±7.99	18.3±8.01
After using the test product for 4 weeks	15.3±7.25*Δ	17.9±7.87

Note: "*" means p<0.05 compared with the product before use, and "Δ" means p<0.05 compared with H.

TEWL is used to characterize the skin barrier function. The higher the value, the worse the skin barrier function, and vice versa. The TEWL value gradually decreased after H2 use, and the decrease was significant at 4 weeks (p<0.05). The TEWL value of H after 4 weeks of use was not significantly different from that before use (P>0.05). After 4 weeks of use, the difference between H2 and H was significant (p<0.05), indicating that H2 has the effect of repairing the skin barrier function.

Example 7 : Preparation of anti-wrinkle eye cream composition

The above anti-wrinkle eye cream composition is prepared as follows:

1. Oil phase: add raw materials 1, 3, 4, and 5 to the oil phase pot, and heat the above-mentioned raw materials to 80°C, dissolve, and mix well;

2. Mix the raw materials of No. 6, 7, 9, 10, 11 at room temperature evenly;

3. Water phase: heat 2 to 80°C, add the mixture of step 3, dissolve, and mix well;

4. Emulsification: add the water phase and the oil phase to the emulsification tank, keep the temperature at 80°C, homogenize and emulsify at 3000rpm for 5 minutes, add No. 12 raw material after the emulsification is completed;

5. Add No. 8 raw material, stir and add the mixture of step 2 when the temperature is lowered to 40°C, stir evenly and then discharge to obtain the anti-wrinkle eye cream composition.

Using the half-face control test method, 20 volunteers were tested as follows before using the product and 4 weeks after using the product:

1) Use primos to take pictures of the volunteers' left and right outer corners of the eyes, and use software to calculate wrinkle parameters, including wrinkle number, wrinkle area, wrinkle depth, etc.;

2) Use Corneometer to test the skin moisture content of the left and right corners of the eyes.

The results showed that 18 of the 20 subjects had a significant increase in water content in the skin of the corners of the eyes, and 16 of them were significantly lighter and lighter, the area of wrinkles and the number of wrinkles decreased.

Example 8 : Preparation of anti-wrinkle emulsion composition

The formula of the anti-wrinkle emulsion composition is as follows:

To	ingredient	weight%
1	4 times concentrated white birch (BETULA ALBA) sap	72.10
2	Dipropylene glycol	6.00
3	glycerin	6.00
4	Polydimethylsiloxane	4.00
5	Cetyl ethylhexanoate	2.00
6	Phenyl trimethicone	1.00
7	PEG-60 Glycerol Isostearate	1.20
8	PEG/PPG-17/6 copolymer	1.00
9	Trehalose	1.00
10	PEG-5 glyceryl stearate	1.00
11	Palmitoyl oligopeptides	1.00
12	Retinol	0.50
13	Phytosterol Macadamia Oleate	1.00
14	Behenyl alcohol	0.80
15	Phenoxyethanol	0.40
16	Tromethamine	0.30
17	Methyl paraben	0.20
18	Acrylic (ester) type/vinyl isodecanoate cross-linked polymer	0.20
19	Allantoin	0.20
20	Xanthan gum	0.10

The aforementioned anti-wrinkle emulsion composition is prepared as follows:

1. Oil phase: Add raw materials No. 4, 5, 6, 7, 10, 12, 13, and 14 to the oil phase pot, and heat to 80°C, dissolve and mix well;

2. Mix the raw materials such as No. 1, 2, 8, 9, 17, and 19 at room temperature; mix the raw materials such as No. 3, 18, and 20 at room temperature;

3. Water phase: heat the mixture of step 2 to 80°C, add the mixture of step 3, dissolve, and mix well;

4. Emulsification: add the water phase and the oil phase to the emulsification tank, keep the temperature at 80°C, homogenize emulsification at 3000rpm for 5 minutes, add No. 16 raw material after emulsification is completed;

5. Add No. 8 raw material, add 11, 12, and 15 raw materials when the temperature is lowered to 40°C under stirring, and discharge after mixing to obtain the anti-wrinkle emulsion composition.

19 subjects used the emulsion of the above formula for 4 weeks, and subjectively evaluated the use. Among them, 18 people reported a significant increase in skin water content, smooth, tender and elastic skin; 15 people reported significant improvement in nasolabial lines and eye wrinkles.

Claims (6)

1. (A) Concentrated birch sap and (B) one or more selected from polypeptides, sodium hyaluronate, retinol, retinal, retinol palmitate, tocopherol, ubiquinone and adenosine The use of the combination of species in an anti-aging cosmetic composition, wherein the concentration ratio of the concentrated birch tree juice is 1.05-8 times, preferably 1.1-4 times, more preferably 1.2-2 times.
2. An anti-aging cosmetic composition comprising (A) concentrated birch sap, the concentration of which is 1.05-8 times, preferably 1.1-4 times, more preferably 1.2-2 times, and (B) selected from polypeptides, transparent One or more of sodium phosphate, retinol, retinal, retinol palmitate, tocopherol, ubiquinone and adenosine.
3. The anti-aging cosmetic composition of claim 2, wherein the content of the concentrated birch sap of the component (A) is 18-98% by weight, preferably 20-95% by weight, more preferably 22-92% by weight, most preferably 30 -90% by weight, based on the total weight of the cosmetic composition.
4. The anti-aging cosmetic composition of claim 2 or 3, wherein the content of the component (B) is 0.005-10% by weight, preferably 0.01-8% by weight, more preferably 0.1-6% by weight, most preferably 0.5-5% by weight Weight, based on the total weight of the anti-aging cosmetic composition.
5. The anti-aging cosmetic composition of any one of claims 2-4, wherein the polypeptide is selected from palmitoyl tripeptide-5, palmitoyl tetrapeptide-7, acetyl hexapeptide-3, dipeptide diaminobutyryl benzyl Amide diacetate, palmitoyl oligopeptide, hexapeptide-9 and any combination thereof.
6. The anti-aging cosmetic composition according to any one of claims 2-5, wherein the anti-aging cosmetic composition does not contain any added water.