Skin care lotion

Abstract

The invention provides a skin care lotion which can be effectively absorbed in a transdermal manner, the skin care functions of tightening, wrinkle resistance and the like are generated after absorption, and the skin care lotion comprises the following components in percentage by mass based on the total weight of the skin care lotion: 20-30% of an oil phase, 20-30% of a water phase, 20-30% of a water phase, 10-30% of a water phase and 10-30% of a water phase, the proportion of the water phase is 60%-70%; the emulsion accounts for 8-12%, and the emulsion contains a yeast extract, a plant extract and other substances capable of achieving any one or more effects of skin whitening, moisturizing, anti-aging, repairing and cell growth.

Description

Technical Field

[0001] The invention belongs to the field of skin care and beauty, and particularly relates to a skin care lotion. Background Art

[0002] Human skin mainly acts as a barrier. This structure helps protect the body from external stimuli, while also preventing the entry of skin care ingredients. Therefore, if skin care products want to enter the skin, they first need to pass through the epidermis and enter the skin to take effect.

[0003] The skin consists of three parts: the epidermis, dermis, and subcutaneous tissue. The epidermis is the shallowest layer of the skin. The epidermis is divided into the stratum corneum, the clear layer, the granular layer, the spinous layer, and the basal layer. The basal layer is connected to the dermis with the help of the basement membrane. The stratum corneum is the outermost layer of the epidermis, which is composed of 5 to 15 layers of keratinocytes and intercellular lipids. This structure is figuratively likened to a "brick wall structure." The thickness of the stratum corneum is 15 to 50 μm, the water content is very low (5% to 20%), and the metabolism is inactive. It is the main barrier to the transdermal absorption of chemical substances. Intercellular lipids are mainly composed of 45% to 50% ceramide, 25% cholesterol, 15% long-chain free fatty acids, and 5% other lipids. The dermis is located below the epidermis and supports the epidermis. It is mainly composed of connective tissue, including collagen fibers, elastic fibers, and matrix. There are also other tissues in the dermis, such as nerves, blood vessels, lymphatic vessels, muscles, hair follicles, sebaceous glands, and sweat glands.

[0004] Transdermal absorption pathways of skin care products

[0005] 1. Intercellular pathway: entering from the gap between cells

[0006] 2. Penetrate cells: enter through the cell membrane

[0007] 3. Enter through the sweat gland pores and other skin appendages

[0008] The physiological structure of transdermal absorption of cosmetics mainly includes the stratum corneum, hair follicles, sebaceous glands and sweat duct openings. There are two ways of penetration through the stratum corneum: ① intercellular pathway: chemical substances bypass keratinocytes and penetrate into the subcutaneous tissue through the intercellular matrix continuously distributed between keratinocytes; ② transcellular entry: chemical substances directly pass through keratinocytes and intercellular matrix, and diffuse alternately in the aqueous phase and lipid phase. ③ The transdermal pathway in which chemical substances directly enter the dermis through skin appendages such as hair follicles, sebaceous glands and sweat duct openings is also called the bypass pathway. Macromolecular substances and ionic substances have difficulty passing through the lipid-rich stratum corneum and may enter the skin through this pathway.

[0009] The main theories of transdermal absorption of cosmetics are: 1. Diffusion theory, 2. Osmotic pressure theory, 3. Hydration theory, 4. Like dissolves like theory and 5. Structural change theory.

[0010] 1. Diffusion theory: Diffusion theory is based on the fact that most chemicals enter the body through the skin by passive diffusion;

[0011] 2. Osmotic pressure theory: The skin is regarded as a semipermeable membrane, which separates solutions with different concentrations. Solutions diffuse from high concentration to low concentration through the semipermeable membrane, thus allowing the active ingredients of cosmetics to be absorbed by the skin.

[0012] 3. Hydration theory: Skin hydration is conducive to transdermal absorption;

[0013] 4. Like dissolves like principle: It is relatively easy to understand that the more similar the composition, structure and characteristics of the chemical ingredients of the skin care ingredients are to the skin, the easier it is to be absorbed through the skin. Therefore, there are many bionic ingredients that simulate the skin barrier components, which use the principle of like dissolves like.

[0014] 5. Structural change theory: It refers to the use of penetration enhancers to reduce the barrier function of the skin, improve the solubility of the stratum corneum, and improve the distribution of chemicals in it, thereby promoting the absorption of chemicals in the skin. This is the principle used to add some penetration enhancers to skin care ingredients.

[0015] In addition, some raw materials used in skin care, such as yeast extract, collagen, elastin, chitosan and some plant extracts, have limited absorption on the skin surface and poor effects. In order to determine whether cosmetics can be effectively absorbed by the skin, in addition to excellent ingredients, they must also promote skin penetration and enhance transdermal absorption so that effective active ingredients can reach the skin and take effect. Therefore, how to develop a skin care lotion that can be effectively absorbed through the skin is a problem that needs to be solved. Summary of the invention

[0016] The invention provides a skin care lotion capable of effective transdermal absorption, which produces skin care functions such as firming and anti-wrinkle after absorption.

[0017] To this end, the present invention provides the following technical solutions.

[0018] The present invention provides a skin care lotion, characterized in that: the composition of the lotion, measured by mass percentage based on the total weight of the skin care lotion, is as follows: the oil phase accounts for 20-30%; the water phase accounts for 60%-70%; the emulsion accounts for 8%-12%, and the emulsion contains yeast extract, plant extract and other substances that can play any one or more of the roles of skin whitening, moisturizing, anti-aging, repairing and cell growth.

[0019] The skin care lotion provided by the present invention also has the following characteristics: wherein, in terms of mass ratio, the composition of the skin care lotion is: oil phase: water phase: emulsifier is: 2-2.5:6-7:1.

[0020] The skin care lotion provided by the present invention also has the following characteristics: the composition of the skin care lotion, measured by mass percentage based on the total weight of the transdermal composition, includes:

[0021] 5%-20% or 10-20% or 13-18% glycerol;

[0022] 5%-20% or 5-15% or 8-12% emulsifier;

[0023] 1%-10% or 3-8% butylene glycol or propylene glycol;

[0024] 0.5-5% or 1-3% laurocapram;

[0025] 0.3%-3% or 1-2% carboxymethyl chitosan;

[0026] 60-70% water;

[0027] 1.5%-37% of any one or more of yeast extract, sodium ascorbyl phosphate, superoxide dismutase, soluble collagen, elastin, hydroxypropyl tetrahydropyrantriol, carnosine, sodium polyglutamate, kelp extract, witch hazel extract, calendula extract, centella asiatica extract, aloe extract and biotin;

[0028] 0.45%-5% of any one or more of tocopherol (vitamin E), retinyl palmitate and flavor.

[0029] The skin care lotion provided by the present invention also has the following characteristics: in terms of mass percentage of the total weight of the transdermal composition, the composition of the skin care lotion comprises:

[0030] 5%-20% or 10-20% or 13-18% glycerol;

[0031] 5%-20% or 5-15% or 8-12% emulsifier;

[0032] 1%-10% or 3-8% butylene glycol or propylene glycol;

[0033] 0.5-5% or 1-3% laurocapram;

[0034] 0.3%-3% or 1%-2% carboxymethyl chitosan;

[0035] 60%-70% water;

[0036] Any one or more of 1%-10% yeast extract, 0.3%-3% sodium ascorbyl phosphate, 0.5%-10% superoxide dismutase, 0.005%-1% soluble collagen, 0.005%-1% elastin, 0.005%-1% hydroxypropyl tetrahydropyrantriol, 0.005%-1% carnosine, 0.005%-1% sodium polyglutamate, 0.01%-2% kelp extract, 0.01%-2% witch hazel extract, 0.01%-2% calendula extract, 0.01%-2% centella asiatica extract, 0.01%-2% aloe extract, and 0.01%-2% biotin;

[0037] Any one or more of 0.3%-3% tocopherol (vitamin E), 0.1%-1% retinyl palmitate and 0.05%-1% flavor.

[0038] The skin care lotion provided by the present invention also has the following characteristics: wherein the emulsifier is selected from any one or more of polysorbate 80, polysorbate 60, polysorbate 20 and sorbitan oleate.

[0039] The skin care lotion provided by the present invention also has the following characteristics: it contains a preservative. Preferably, the preservative is contained in an amount of 0.1-1% by weight based on the total weight of the skin care lotion. Furthermore, the preservative is selected from any one or more of phenoxyethanol, methylparaben, ethylparaben, propylparaben, benzyl alcohol and sorbic acid.

[0040] The present invention also provides the use of the aforementioned skin care lotion in achieving skin care effects, wherein, when used, it is used transdermally by any one or more of applying, transdermal instrumentation, sticking, and spraying.

[0041] The present invention also provides the use of the aforementioned skin care emulsion in the preparation of a medicine or product for achieving skin care effects.

[0042] Function and Effect of the Invention

[0043] The skin care lotion provided by the present invention has an appropriate proportion of the oil phase, the water phase and the emulsion in its composition, forms an oil-in-water microemulsion with nanometer average particle size, has a very good transdermal effect, and can achieve skin care functions such as firming and anti-wrinkle. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 The standard curve of the BSA standard in Example 4 was fitted according to four parameters;

[0046] Figure 2 This is the toe-closing phenotype of the mouse experiment in Example 5. DETAILED DESCRIPTION

[0047] The specific implementation of the present invention is described below in conjunction with the accompanying drawings. For the specific methods or materials used in the embodiments, those skilled in the art can make conventional replacement selections based on the technical ideas of the present invention and according to existing technologies, and are not limited to the specific description of the embodiments of the present invention.

[0048] Unless otherwise specified, the methods used in the examples are all conventional methods; the materials, reagents, etc. used, unless otherwise specified, can be obtained from commercial sources.

[0049] Emulsion refers to a non-uniformly dispersed liquid preparation formed by two immiscible liquid phases, one of which is dispersed in the other liquid phase in the form of small droplets. The phase that forms the droplets is called the dispersed phase, internal phase or non-continuous phase, and the other liquid phase is called the dispersion medium, external phase or continuous phase.

[0050] Emulsions are composed of an oil phase (represented by O), an emulsifier, and a water phase (represented by W). Depending on the type, properties, and phase volume ratio (φ) of the emulsifier, an oil-in-water (O / W) or water-in-oil (W / O) emulsion or a composite emulsion can be formed.

[0051] Aqueous phase: water or aqueous solution. Aqueous solution refers to the dissolution of water-soluble raw materials in water;

[0052] Oil phase: refers to the raw materials that are not easily soluble in water being dissolved in oily substances.

[0053] According to the size of the emulsion particles, that is, the size of the emulsion droplets, emulsions are classified into ordinary emulsions, submicron emulsions and nanoemulsions:

[0054] 1. Ordinary milk: The droplet size of ordinary milk is generally 1μm-100μm, and it is a milky white opaque liquid.

[0055] 2. Subemulsions: The droplet size is generally 0.1μm-1.0μm, and submicron emulsions are often used as carriers for gastrointestinal administration. Intravenous emulsions should be submicron emulsions, and the particle size is generally in the range of 0.25μm-0.4μm.

[0056] 3. Nanoemulsions: The size of the emulsion droplets is <100nm, generally in the range of 10-100nm. Nanoemulsions are also called microemulsions.

[0057] As a natural molecular alkaline polysaccharide, chitosan has film-forming, antibacterial, anti-inflammatory, hemostatic, analgesic, antipruritic, scar-removing, wound healing and anti-oxidation effects. Chitosan has a good inhibitory effect on microorganisms such as bacteria, yeast, and fungi. It has a significant inhibitory effect on bacteria present in the human epidermis, such as Staphylococcus epidermidis, Escherichia coli, and tropical Candida albicans, as well as Pseudomonas aeruginosa, Staphylococcus aureus, and suppurative Staphylococcus aureus infections that are prone to occur in burn patients. Chitosan has strong moisturizing properties, and its moisturizing ability even exceeds that of sodium hyaluronate. It is known as a "super moisturizer". Chitosan can absorb heavy metals and residues in cosmetics. Chitosan has a repairing power and can repair the skin barrier. It is useful for sensitive skin and acne skin.

[0058] The main application effects of yeast extract are skin rejuvenation, anti-aging and water-locking and moisturizing. Yeast extract has a good skin rejuvenation effect, which is achieved through three main modes of action: (1) reducing the transfer of produced melanin to surface cells; (2) promoting the metabolism of melanin-containing keratinocytes to accelerate keratin shedding; (3) promoting the synthesis of epidermal proteins to regulate skin texture. Yeast extract has a good free radical scavenging and anti-aging effect, which can regulate skin texture, fine lines and wrinkles, increase skin elasticity, make the skin smooth and delicate, and can also reduce and prevent the dull skin color caused by the early aging process. Yeast extract also has a strong water-locking effect: it is rich in natural moisturizing factors, which can enable the stratum corneum to retain moisture more effectively, give the skin deep moisture and prevent skin dryness; it can strengthen the skin barrier function by increasing the content of ceramide in the stratum corneum and reduce transepidermal water loss; it can also repair the damaged stratum corneum lipid barrier, improve skin resistance and regulate skin condition.

[0059] The main applications of superoxide dismutase in cosmetics include: (1) as a cosmetic additive, it can prevent skin aging and have a skin care effect; (2) prevention and treatment of related skin diseases, which have been widely used abroad. At present, many high-end cosmetics at home and abroad have added SOD and made it into facial masks, lotions, and creams.

[0060] Soluble collagen is a natural protein that has important functions in the skin. Collagen maintains the elasticity and firmness of the skin, but as we age, the amount of collagen in the skin decreases, causing sagging and wrinkles. Soluble collagen, as a form that can penetrate deeper into the skin, can replenish collagen in the skin and improve the condition of the skin. Soluble collagen can stimulate and promote collagen synthesis, help repair damaged collagen, and promote the production of new collagen, thereby improving the elasticity and firmness of the skin and improving wrinkles and fine lines.

[0061] Elastin is an important component of the skin and the basis of skin youth. Elastin peptides have the effects of promoting skin damage repair, blood vessel regeneration and fibroblast proliferation. It can also enhance the elasticity of human skin, making the skin smooth and elastic.

[0062] Vitamins are essential organic substances for the human body. Vitamin A can promote the proliferation of epidermal cells, increase the formation of dermal collagen and elastin, and has anti-aging effects. Vitamin E is a natural antioxidant that can promote metabolism and improve skin elasticity. Vitamin C has strong antioxidant properties and can promote collagen synthesis and inhibit collagen decomposition. Vitamin C and vitamin E have a synergistic effect of scavenging free radicals.

[0063] In addition to the vitamins essential to the human body, there are currently a variety of natural plant active ingredients that have been proven to have anti-wrinkle and firming functions, such as Centella asiatica extract, Osmanthus algae extract, Calendula officinalis extract, etc.

[0064] The skin care lotion provided by the present invention has the following compositions, measured by mass percentage based on the total weight of the skin care lotion: the oil phase accounts for 20-30%; the water phase accounts for 60%-70%; the emulsion accounts for 8%-12%, and the emulsion contains yeast extract, plant extract and other substances that can play any one or more of the roles of skin whitening, moisturizing, anti-aging, repairing and cell growth.

[0065] The composition of the skin care lotion provided by the present invention is shown in Table 1:

[0066]

[0067] Table 1 refers to the composition of the skin care emulsion of the present invention, which is divided into three types: oil phase, water phase and emulsifier. The "yeast extract, plant extract and other substances that can play any one or more of the roles of skin whitening, moisturizing, anti-aging, repair, and cell growth" contained therein may be oily (soluble in oil) or aqueous (soluble in water).

[0068] In one example, the skin care lotion of the present invention satisfies the requirements shown in Table 2:

[0069]

[0070] In one example, the composition of the skin care lotion of the present invention is shown in Table 3:

[0071]

[0072] Preferably, the example of Table 3 is composed of Table 4:

[0073]

[0074] In one example, in Tables 3 and 4, the 1.5%-37% aqueous phase composition mentioned therein is specifically: 1%-10% yeast extract (preferably 6-10%), 0.3%-3% sodium ascorbyl phosphate (preferably 1-3%), 0.5%-10% superoxide dismutase (preferably 0.5-2%), 0.005%-1% soluble collagen, 0.005%-1% elastin (preferably 0.005%-0.01%), 0.005%-1% hydroxypropyl tetrahydropyrantriol (preferably 0.005%-0.01%), 0.005%-1% carnosine (preferably 0.005%-0.01%). %), 0.005%-1% sodium polyglutamate (preferably 0.005%-0.01%), 0.01%-2% of Osmanthus algae extract (preferably 0.01%-0.03%), 0.01%-2% witch hazel extract (preferably 0.01%-0.03%), 0.01%-2% calendula extract (preferably 0.01%-0.03%), 0.01%-2% Centella asiatica extract (preferably 0.01%-0.03%), 0.01%-2% aloe extract (preferably 0.01%-0.03%) and 0.01%-2% biotin (preferably 0.01%-0.03%).

[0075] In one example, in Tables 3 and 4, the 0.45%-5% oil phase mentioned therein is, specifically, any one or more of: 0.3%-3% tocopherol (vitamin E) (preferably 1-3%), 0.1%-1% retinol palmitate (preferably 0.1%-0.5%) and 0.05%-1% flavor (preferably 0.1%-0.03%).

[0076] In one example, the emulsifier is selected from any one or more of polysorbate 80, polysorbate 60, polysorbate 20 and sorbitan oleate.

[0077] In one example, the skin care lotion also contains a preservative, preferably, in terms of the mass percentage of the total weight of the transdermal microemulsion system: 0.1-1% of the preservative. Further, the preservative is selected from any one or more of phenoxyethanol, methylparaben, ethylparaben, propylparaben, benzyl alcohol and sorbic acid.

[0078] The average particle size of the particles of the skin care emulsion of the present invention is 10-15 nm.

[0079] The skin care lotion of the present invention is used in achieving skin care effects. When used, it can be used transdermally by any one or more of smearing, transdermal instruments, sticking and spraying. When used, it can be used transdermally by any one or more of smearing, transdermal instruments, sticking and spraying.

[0080] The skin care emulsion of the present invention can also be used in preparing medicines or products with skin care effects.

[0081] In addition, in the present invention, water is preferably any one or more combinations of distilled water, drinking water, sterile water, ultrapure water and deionized water.

[0082] The following describes the method in conjunction with specific embodiments.

[0083] Example 1

[0084] The formula provided in this embodiment has the same ratio range of oil, water and emulsion as the skin care lotion of the present invention, except that the macromolecular active ingredient botulinum toxin is added therein to prove that the ratio of the present invention has good transdermal efficiency, as shown in Table 5:

[0085]

[0086] Preparation method:

[0087] (1) Using an electronic balance, weigh purified water, carboxymethyl chitosan, and botulinum toxin A in the formula into a 3 L beaker 1, and stir at room temperature to dissolve (rotation speed: 400-500 rpm).

[0088] (2) Using an electronic balance, weigh polysorbate 80, laurocapram, phenoxyethanol, propylene glycol and glycerol in order according to the formula into a 2 L beaker 2, and stir evenly at room temperature (speed: 250-300 rpm).

[0089] (3) Add the solution in beaker 2 to beaker 1 and stir evenly at room temperature (speed: 400-500 rpm).

[0090] (4) Add purified water to the solution prepared in step 3) to make up to 1000 g, and stir at room temperature to mix.

[0091] Example 2

[0092] The formula of the skin care lotion provided in this embodiment is shown in Table 6.

[0093]

[0094] Preparation method:

[0095] (1) Using an electronic balance, weigh purified water, carboxymethyl chitosan, yeast extract, superoxide dismutase (SOD) and sodium ascorbyl phosphate in accordance with the formula into a 3 L beaker 1, and stir at room temperature to dissolve (rotation speed: 400-500 rpm).

[0096] (2) Using an electronic balance, weigh polysorbate 80, tocopherol (vitamin E), retinyl palmitate, phenoxyethanol, (daily use) fragrance, butylene glycol and glycerin in order according to the formula into a 2 L beaker 2, and stir evenly at room temperature (speed: 250-300 rpm).

[0097] (3) Add the solution in beaker 2 to beaker 1 and stir evenly at room temperature (speed: 400-500 rpm).

[0098] (4) Add purified water to the solution prepared in step (3) and make up to room temperature.

[0099] Stir to mix.

[0100] Example 3

[0101] The formula of the skin care lotion provided in this embodiment is shown in Table 7.

[0102]

[0103]

[0104] Preparation method:

[0105] (1) Using an electronic balance, weigh purified water, hydroxypropyl tetrahydropyrantriol, carnosine, sodium polyglutamate, biotin, kelp extract, calendula extract, Centella asiatica extract, aloe extract, carboxymethyl chitosan, sodium ascorbyl phosphate, yeast extract, superoxide dismutase (SOD), soluble collagen, and elastin according to the formula amount into a 3 L beaker 1, and stir at room temperature to dissolve (rotation speed: 400-500 rpm).

[0106] (2) Using an electronic balance, weigh polysorbate 80, tocopherol (vitamin E), retinyl palmitate, phenoxyethanol, (daily use) fragrance, butylene glycol and glycerin in order according to the formula into a 2 L beaker 2, and stir evenly at room temperature (speed: 250-300 rpm).

[0107] (3) Add the solution in beaker 2 to beaker 1 and stir evenly at room temperature (speed: 400-500 rpm).

[0108] (4) Add purified water to the solution prepared in step (3) to make up to 1000 g, and stir at room temperature to mix.

[0109] The preparations obtained in Examples 1-3 have good emulsification effect, high transparency, and no stratification. The average particle size detected by a nanoparticle size potentiometer is 10-15 nm, and the average particle size of the preparation obtained in Example 3 is 12.99 nm.

[0110] Comparative Example 1

[0111] The formula of this comparative example is shown in Table 8

[0112]

[0113] Preparation method:

[0114] (1) Add the acid ester to a 2L beaker and stir for 5 minutes; weigh oleyl alcohol and oleic acid according to the formula using an electronic balance and add them to the beaker, stir for 5 minutes; weigh laurocapram and phenoxyethanol according to the formula using an electronic balance and add them to the beaker, stir evenly at room temperature (speed: 200-250rpm, 5min). (2) Use an electronic balance to weigh propylene glycol and menthol according to the formula and add them to a blue-capped bottle in sequence, heat to 60°C (about 10 minutes) to fully dissolve, then add them to the solution obtained in step 1), and stir evenly at room temperature (speed: 200-250rpm, 5min).

[0115] (3) Add 10 mg of botulinum toxin A to the solution prepared in (2), and stir at room temperature (speed: 200-250 rpm, 5 min).

[0116] (4) Add purified water to the solution prepared in step (3) to make up to 1000 g, and stir at room temperature to mix.

[0117] Comparative Examples 2-10

[0118] The formulas of Comparative Examples 2-10 are shown in Table 9, calculated by mass percentage:

[0119]

[0120] According to the respective formulations in Comparative Examples 2-10, the oil phase is fully mixed and dissolved, the water phase is fully mixed and dissolved, and then the phases are mixed to obtain the products. The specific preparation methods are all conventional techniques and can be selected by referring to the previous embodiments, and will not be described one by one.

[0121] The appearance effects of the prepared comparative examples 2-10 are not as good as those of the examples 1-3, as shown in Table 10:

[0122]

[0123] Example 4

[0124] This experiment was used to detect the transdermal efficiency.

[0125] The botulinum toxin A added in Example 1 and Comparative Example 1 was replaced with BSA protein. The BSA protein emulsion prepared in Example 1 was 1.0 mg / ml, and the BSA protein emulsion prepared in Comparative Example 1 was 0.62 mg / ml. The transdermal experiment was carried out using the Franz diffusion cell method.

[0126] Test equipment and materials

[0127] 1. Reagents

[0128] Phosphate buffer, protein standards, test products, protein detection kit (ultra-high sensitivity);

[0129] 2. Consumables

[0130] In vitro pig skin, centrifuge tubes, EP tubes, pipette tips, pre-coated microplates for transdermal evaluation; 3. Equipment

[0131] TP-6 transdermal diffusion instrument (Tianjin Jingtuo Instrument Technology Co., Ltd.), multifunctional microplate reader (Thermoscientific, Varioskan Lux), medical refrigerator (Qingdao Haier Biomedical Co., Ltd.), biochemical incubator (Shanghai Yiheng Scientific Instrument Co., Ltd.), and pipette (Eppendorf AG).

[0132] 1. Preparation before the experiment:

[0133] (1) The pig skin required for the experiment was taken out of the -20°C refrigerator and thawed at 4°C, then washed with physiological saline and set aside.

[0134] (2) Before the experiment, clean the supply chamber, diffusion chamber (receiving chamber) and stirring bar.

[0135] (3) Add pure water to the water tank, preferably flush with the diffusion cell frame, and set the temperature to 32°C and the rotation speed.

[0136] (4) After the temperature of the TP-6 ​​transdermal diffusion instrument rises to the set temperature, the pig skin is placed between the supply chamber and the diffusion chamber. Stir at a constant speed to keep the solution isothermal and the concentration of the exudate uniform.

[0137] 2. Experimental methods:

[0138] (1) Set up three experimental groups and one blank group.

[0139] (2) Adding BSA protein test solution: Add 150 μl (1 mg / ml) of the BSA protein emulsion test solution prepared in Example 1, 150 μl (0.62 mg / ml) of the BSA protein emulsion test solution prepared in Comparative Example 1, and 150 μl (1 mg / ml) of the BSA protein test solution dissolved in PBS to the first three supply chambers, respectively; add 150 μl of PBS to the blank group.

[0140] (3) Fill the receiving chamber with PBS, take four 1.5 ml EP tubes, and take 400 μl of the liquid in the receiving chamber from the sampling port every hour for subsequent OD detection, and mark the sampling time and experimental group number. The total sampling time is 6 hours. After sampling, each receiving chamber must be supplemented with 400 μl PBS to maintain the volume of the receiving chamber unchanged.

[0141] (4) All collected samples were placed in a 4°C refrigerator for later use.

[0142] 3. ELISA test:

[0143] 1. Reagent preparation:

[0144] a. Before use, equilibrate all reagents to room temperature (18-25°C). Set the detection wavelength to 450nm according to the instructions of the microplate reader and preheat for 15 minutes before reading the plate.

[0145] b. Wash Buffer: Dilute 20 ml of concentrated wash buffer with 580 ml of deionized water to prepare 600 ml of wash buffer.

[0146] c. Standard working solution: First, centrifuge the standard at 1000g for 1 min, add 2ml of standard sample diluent, and mix it with the standard sample diluent. Pipette 300μl of 20000ng / ml standard diluent into the first tube and mix it to make 6666.7ng / ml working solution. Then transfer 300μl of solution from the front tube to the back tube. Mix each tube thoroughly before the next transfer. Set 6 points of standard dilution, namely 20000ng / ml, 6666.7ng / ml, 2222.2ng / ml, 740.7ng / ml, 246.9ng / ml, 0ng / ml.

[0147] d. Detection Reagent A working solution: Calculate the required amount (100 μl / well) before the experiment. During the preparation, you should prepare 100-200 μl more than the calculated amount. Centrifuge the stock solution tube slightly before use, and dilute the 100× concentrated detection reagent A to 1× working solution A with detection diluent A (e.g.: 10 μl detection reagent A + 990 μl detection reagent A diluent).

[0148] e. Detection Reagent B working solution: Calculate the required amount (100μl / well) before the experiment. During the preparation, you should prepare 100-200μl more than the calculated amount. Centrifuge the stock solution tube slightly before use, and dilute the 100× concentrated detection reagent B to 1× working solution B with detection diluent B (e.g.: 10μl detection reagent B + 990μl detection reagent B diluent).

[0149] 2. Experimental Methods

[0150] (1) Set up standard wells, sample wells, and blank wells respectively. Add six standards of different concentrations (including zero well, 50 μL / well) in sequence, add 50 μL of the sample to be tested (the sample collected from the sampling port of the receiving chamber) to the other wells, and then immediately add the detection reagent A working solution, 50 μL / well, gently shake to mix, cover with the sealing film provided by the kit, and incubate at 37°C for 60 minutes.

[0151] (2) Discard the liquid in all wells, add 350uL of washing buffer to each well, soak for 60 seconds, then pour out the liquid in each well and pat dry on clean absorbent paper. Repeat this washing step for a total of 3 times.

[0152] (3) Add 100 μL of detection reagent B working solution to each well, cover with sealing film, and incubate at 37°C for 30 minutes.

[0153] (4) Discard the liquid in each well and repeat the washing process in step 2 5 times.

[0154] (5) Add 90 μL of TMB reagent to each well, cover with a new sealing film, and incubate at 37°C in the dark for 10-20 minutes.

[0155] (6) Add 50 μL of stop solution to each well in the same order as adding the color development solution, and gently shake to mix.

[0156] (7) Ensure that there are no bubbles or water mist at the bottom of the ELISA plate wells, and immediately measure the absorbance OD value of each well at 450 nm and record the ELISA reader reading.

[0157] 4. Test data and conclusions

[0158] (1) The BSA standard was fitted with a four-parameter standard curve as follows: Figure 1 shown.

[0159] Standard curve drawing:

[0160] Process the OD value data of the detected standard protein:

[0161] The data were fitted with four parameters to obtain the standard curve. The four-parameter fitting formula is:

[0162]

[0163] For specific standard curves, see Figure 1 .

[0164] (2) Sample test results

[0165] Table 11 is the BSA concentration permeating through the skin of different test samples at different time periods calculated based on the standard curve.

[0166]

[0167]

[0168] Based on the results, permeation calculation is performed:

[0169] (1) After 24 hours, the BSA emulsion protein test solution prepared in Example 1 had a concentration of 239.6 ng / ml in the receiving chamber, and its volume was 15 ml. The BSA concentration in the donor chamber was 1 mg / ml, and the volume added to the donor chamber was 150 ul. Based on this, it was calculated that the transdermal efficiency of the BSA emulsion prepared in Example 1 was 2.40%.

[0170] (2) Comparative Example 2 prepared a BSA emulsion protein test solution. After 24 hours, the concentration in the receiving chamber was 147.1 ng / ml, and its volume was 15 ml. The BSA concentration in the supply pool was 0.62 mg / ml, and the volume added to the supply chamber was 150 ul. Based on this, it was calculated that the transdermal efficiency of the BSA emulsion prepared in Example 5 was 2.37%.

[0171] (3) After 24 hours, the BSA protein test solution dissolved in PBS had a concentration of 125.4 ng / ml in the receiving chamber, which had a volume of 15 ml, while the BSA concentration in the donor chamber was 1 mg / ml, and the volume added to the donor chamber was 150 ul. The transdermal efficiency of PBS was calculated to be 1.25%.

[0172] It can be seen that, compared with the control, the transdermal effects of Example 1 and Comparative Example 2 are very good, and the transdermal effect of Example 1 is better than that of Comparative Example 2.

[0173] Example 5

[0174] The efficacy of botulinum toxin A microemulsions prepared in Example 1 and Comparative Example 1 at a concentration of 10 μg / ml was verified in mice.

[0175] In order to compare the difference in transdermal efficiency of macromolecular active substances between Example 1 and Comparative Example 1, we used 17-19g SPF CD-1 (ICR) mice, anesthetized with tribromoethanol, shaved the skin of the right hind leg, cleaned the smeared area with a medical cotton swab dipped in 10% alcohol before each application, and then applied the medicine after drying. 200μl of botulinum toxin A microemulsion was applied to the skin of the right hind leg of the mouse every day, once a day, for 7 consecutive days. The left hind leg of the mouse was not applied with botulinum toxin A.

[0176] The experimental results show that 2 to 3 days after the mice were coated with botulinum toxin A, the toes of the right hind leg showed a phenotype of being closed (2-5 toes closed together). The experimental results of Example 1 and Comparative Example 1 are shown in Table 12. The emulsion prepared in the formula of Example 1 showed syndactyly at D2: 33% of the experimental mice had two toes closed, and 33% of the experimental mice had five toes closed; D3 syndactyly: 33% of the experimental mice had two toes closed, and 33% of the experimental mice had five toes closed; D4 syndactyly: 66% of the experimental mice had three toes closed, and 33% of the experimental mice had five toes closed; D5 syndactyly: 100% of the experimental mice had five toes closed.

[0177] The emulsion prepared in the formula of Comparative Example 1 showed no syndactyly at D2; syndactyly at D3: 33% of the experimental mice had two syndactyly; syndactyly at D4: 66% of the experimental mice had two syndactyly, and 33% of the experimental mice had three syndactyly; syndactyly at D5: 33% of the experimental mice had three syndactyly, and 66% of the experimental mice had four syndactyly.

[0178]

[0179]

[0180] The results in Table 12 show that the efficacy test results of the botulinum toxin A microemulsion prepared in Example 1 in mice are significantly better than those in Comparative Example 1. The microemulsion of Example 1 not only takes effect earlier, but also has better efficacy than the microemulsion of Example 1. The phenotype of toes close together in Example 1 and Comparative Example 1 is as follows Figure 2 shown.

[0181] According to literature reports, the toes of mice will appear to be close together after botulinum toxin A is injected into the gastrocnemius muscle.

[0182] It can be seen that through the emulsion system we developed, we can make botulinum toxin A into a microemulsion and apply it on the skin of mice, which also achieved the syndactyly phenotype after injection of botulinum toxin A. It was proved in mice that for large molecular active substances, the emulsion delivery system we developed has a very good transdermal effect. It can be seen that our emulsion has a very good transdermal effect.

[0183] Example 7

[0184] The efficacy of the anti-wrinkle and firming cosmetic microemulsion prepared in Example 3 was tested on the human body.

[0185] 1. Under normal circumstances, adult subjects use the product continuously for 28 days according to the instructions to evaluate whether the product has the effects of moisturizing, repairing, firming, and anti-wrinkle, and whether the product is suitable for sensitive skin and is mild and non-irritating.

[0186] 2. Subjects: A total of 31 valid subjects completed the assessment, including healthy Chinese women with sensitive skin (SGS (SGS Standards Technical Services Co., Ltd.) internal sensitive skin questionnaire screening), aged 31 to 60 years old, with an average age of 52.65±6.08 years old, who met the voluntary inclusion and exclusion criteria of the subjects. The assessment area was the face.

[0187] Subject inclusion and exclusion criteria:

[0188] The subjects for this evaluation were screened from the CPCH efficacy laboratory subject information database of SGS, and healthy subjects who met the following inclusion and exclusion criteria were selected.

[0189] (1) Selection criteria

[0190] Healthy women, age: 28-60 years old;

[0191] Asian (Chinese);

[0192] The subjects had sensitive facial skin (screened by the SGS internal sensitive skin questionnaire);

[0193] The subjects rated their facial discomfort symptoms as 4-7 points (0-9 scale, self-assessment questionnaire) such as (non-persistent) itching or tingling;

[0194] The subjects believed that their facial skin was dull, sagging, and lacked elasticity;

[0195] Forehead wrinkles visual score 3-6, (according to SGS internal atlas);

[0196] The visual score of the outer corner of the eye wrinkles on one random side is 3-6 (according to the internal atlas of SGS); the visual score of the nasolabial wrinkles (nasolabial folds) on one side is 3-6 (according to the internal atlas of SGS);

[0197] Transepidermal water loss (TEWL) > 15 g / h / m2 on one cheek area;

[0198] The stratum corneum moisture content of one cheek area was measured to be <60 a.u.;

[0199] The facial skin has no obvious skin lesions, scars, hair, etc.;

[0200] Able to cooperate well with the assessment project as per the requirements of the plan and maintain a regular life during the study period.

[0201] (2) Exclusion criteria

[0202] Anyone with any of the following conditions must be excluded from this study:

[0203] Those who do not agree to sign the informed consent form;

[0204] Those who are unwilling to comply with the requirements of the program;

[0205] concurrently participate in any other clinical research investigators;

[0206] Evaluate the use of cosmetics and / or medicines on the day;

[0207] She reported being pregnant and breastfeeding;

[0208] were receiving medication during the study period;

[0209] Subjects with infectious skin diseases or atopic dermatitis;

[0210] Those with skin abnormalities such as moles, telangiectasia, etc. at the assessment site;

[0211] Subjects who had received skin peeling or skin treatment within 3 months before participating in the assessment;

[0212] Subjects who had received immunosuppressant therapy within 3 months before participating in the assessment;

[0213] Subjects who had received systemic steroid therapy or phototherapy within 1 month before participating in the assessment;

[0214] Use of topical medications and / or special-effect cosmetics (claiming to be moisturizing, repairing, firming, anti-wrinkle) on the affected area within 2 weeks before the assessment;

[0215] There are lesions, obvious marks, or other abnormalities in the evaluation area, making it difficult to measure;

[0216] Subjects with severe reactions or allergies to cosmetics, medications, or general light exposure;

[0217] In addition to the above, when the project leader determines that it is not appropriate to conduct the evaluation.

[0218] 3. Usage: After cleansing, thoroughly remove oil and keep the skin dry. Take an appropriate amount of test product and drop two drops into the dropper, apply to the corners of the eyes, forehead, nasolabial folds, lower jaw and other wrinkled areas, and massage until the essence (test product) is absorbed. It is recommended to use it every day for 7 days a week. It is recommended to use it for 4 weeks as a cycle (please keep the product in a refrigerator at a constant temperature of 2-8℃ when not in use).

[0219] 4. Evaluation period: before using the product (D0), 14 days after using the product (D14), and 28 days after using the product (D28).

[0220] 5. Evaluation parameters:

[0221] (1) Image acquisition

[0222] Primos CR is used for facial image acquisition, skin wrinkle analysis and photo analysis. A smaller analysis value indicates that skin wrinkles have improved.

[0223] (2) Skin elasticity

[0224] Skin elasticity tester MPA580 is used to test skin elasticity. The increase in the measured values ​​of R2, R5, and R7 indicates that the skin elasticity has improved.

[0225] (3) Skin tightening

[0226] Skin elasticity tester MPA580 is used to detect skin firmness. A smaller F4 measurement value indicates that skin firmness has improved.

[0227] (4) Water content of the stratum corneum

[0228] Skin moisture content meter CM825 is used to detect the moisture content of the stratum corneum of the skin. The larger the measured value, the higher the moisture content of the stratum corneum of the skin.

[0229] (5) Transepidermal water loss rate

[0230] Skin water loss meter TM Hex is used to detect the transepidermal water loss rate of the skin. A smaller measurement value indicates that the skin barrier has improved.

[0231] (6)TC value

[0232] The TC value is a secondary parameter in non-invasive testing, which indicates the amount of water lost through the epidermis per unit area per unit time. A smaller analysis value indicates that the skin barrier function has improved.

[0233] (7) Skin moisture distribution map

[0234] The MoistureMap MM 200 is a unique instrument based on capacitive imaging to observe hydration distribution and texture characteristics. MGL represents the mean grayscale value of skin moisture distribution, and the smaller the value, the higher the moisture content.

[0235] (8) Skin glossiness

[0236] Skin gloss meter GL200 is used to detect skin glossiness. The larger the measured value, the greater the skin glossiness.

[0237] (9) Subject self-assessment

[0238] Subjects self-assessed their usage.

[0239] 6. Evaluation plan design:

[0240] (1) Before using the product (D0):

[0241] After arriving at SGS, the subjects cleaned their faces with facial cleansing products and dried their skin with dry, dandruff-free facial tissues. They sat in a laboratory at 21±1°C and 50±10% RH for 30 minutes and were visually evaluated by dermatologists. Those who met the inclusion criteria entered the next step of evaluation;

[0242] The facial skin stratum corneum moisture content (Corneometer) and transepidermal water loss rate (Tewameter) were measured by laboratory technicians, and those who met the inclusion criteria entered the next step of evaluation;

[0243] The laboratory technicians collected Primos CR images, Moisture Map MM200, Cutometer, and Glossymeter measurements from eligible subjects.

[0244] (2) Subjects fill out questionnaires

[0245] The laboratory technicians explain the use of the products to the subjects and distribute the products. The subjects try the samples on site under the supervision of the laboratory technicians. If any adverse reactions occur, they need to fill in the usage log in time.

[0246] (3) 14 days after using the product (D14)

[0247] After arriving at SGS, the subjects cleaned their faces with facial cleansing products and dried their skin with dry facial tissues. They sat in a laboratory at 21±1℃ and 50±5%RH for 30 minutes. Laboratory technicians collected Primos CR images of the subjects' faces and measured values ​​of facial skin instruments such as Cutometer, Moisture Map MM200, Tewameter, Corneometer, and Glossymeter.

[0248] Laboratory technicians weigh products and check product usage logs

[0249] Subject leaves SGS.

[0250] (4) 28 days after using the product (D28)

[0251] After arriving at SGS, the subjects cleaned their faces with facial cleansing products and dried their skin with dry facial tissues. They sat in a laboratory at 21±1℃ and 50±5%RH for 30 minutes. Laboratory technicians collected Primos CR images of the subjects' faces and measured values ​​of facial skin instruments such as Cutometer, Moisture Map MM200, Tewameter, Corneometer, and Glossymeter.

[0252] The subjects filled out the questionnaire;

[0253] Laboratory technicians weigh and recycle products and check and recycle product usage logs;

[0254] Subject leaves SGS.

[0255] (5) Data statistics: SPSS 28.0 was used to analyze the data. The evaluation data were tested for normal distribution. If the evaluation data were normally distributed, the T test method was used for statistical analysis. If the evaluation data were not normally distributed, the rank sum test method was used for statistical analysis. The rank sum test was used for statistical analysis of graded data. The statistical method significance level was P < 0.05.

[0256] The product was used continuously for 28 days by 31 healthy Chinese female subjects with sensitive skin. The evaluation results showed that under the evaluation conditions, the product had moisturizing, repairing, firming, and anti-wrinkle effects in 14 days, and was suitable for sensitive skin and mild and non-irritating. The specific results are as follows:

[0257] (1) Instrument evaluation results (Table 13)

[0258]

[0259]

[0260]

[0261] (2) Subjects’ self-assessment (satisfaction)

[0262] After using the product for 14 days, 94% of the subjects felt that the skin was deeply hydrated, 94% of the subjects felt that the skin was hydrated and plump, 94% of the subjects felt that the moisturizing effect was good, 90% of the subjects felt that the skin was firmer, 87% of the subjects felt that the skin became elastic, 87% of the subjects felt that wrinkles were improved, 87% of the subjects felt that fine lines were improved, 87% of the subjects felt that the overall skin condition was improved, 90% of the subjects felt that the product was suitable for sensitive skin, and 100% of the subjects felt that the product was mild and non-irritating;

[0263] After using the product for 28 days, 100% of the subjects felt that it was deeply hydrated, 100% of the subjects felt that their skin was hydrated and plump, 100% of the subjects felt that the moisturizing effect was good, 100% of the subjects felt that their skin was firmer, 97% of the subjects felt that their skin became elastic, 94% of the subjects felt that wrinkles were improved, 97% of the subjects felt that fine lines were improved, 90% of the subjects felt that the overall condition of the skin was improved, 100% of the subjects felt that the product was suitable for sensitive skin, 100% of the subjects felt that the product was gentle and non-irritating, 100% of the subjects were satisfied with the overall effect / efficacy of the product, and 100% of the subjects were willing to continue using the product.

[0264] Evaluation conclusion:

[0265] The product was used continuously for 28 days by 31 healthy Chinese female subjects with sensitive skin. The evaluation results showed that under the evaluation conditions, the emulsion preparation of Example 3 has the effects of moisturizing, repairing, firming and anti-wrinkle, and is suitable for sensitive skin and is mild and non-irritating.

[0266] In summary:

[0267] The transdermal efficiency experiment of BSA confirmed that the emulsion obtained by the specific ratio of the oil phase, the water phase and the emulsifier provided by the present invention can effectively achieve transdermal penetration of macromolecular substances, that is, our emulsion has a good transdermal effect;

[0268] Furthermore, experiments using botulinum toxin on animals confirmed that the emulsion obtained by the specific ratio of oil phase, water phase and emulsifier does have a very good transdermal effect on macromolecular substances, which further shows that our emulsion has a very good transdermal effect;

[0269] Experiments on human skin use have also proven that the skin care lotion provided by the present invention has a very good transdermal effect on the active ingredients of skin care.

Claims

1. A skin care lotion, characterized in that: Measured by mass percentage based on the total weight of the skin care lotion, the composition of the lotion is: The oil phase accounts for 20-30%; The proportion of water phase is: 60%-70%; Emulsion accounts for: 8%-12%, The emulsion contains yeast extract, plant extract and other substances that can play any one or more roles in skin whitening, moisturizing, anti-aging, repairing and cell growth.

2. The skin care lotion according to claim 1, characterized in that: in, By mass ratio, the composition of the skin care lotion is: The ratio of oil phase: water phase: emulsifier is: 2-2.5:6-7:

1.

3. The skin care lotion according to claim 1 or 2, characterized in that: Measured by mass percentage of the total weight of the transdermal composition, the composition of the skin care lotion includes: 5%-20% or 10-20% or 13-18% glycerol; 5%-20% or 5-15% or 8-12% emulsifier; 1%-10% or 3-8% butylene glycol or propylene glycol; 0.5-5% or 1-3% laurocapram; 0.3%-3% or 1-2% carboxymethyl chitosan; 60-70% water; 1.5%-37% of any one or more of yeast extract, sodium ascorbyl phosphate, superoxide dismutase, soluble collagen, elastin, hydroxypropyl tetrahydropyrantriol, carnosine, sodium polyglutamate, kelp extract, witch hazel extract, calendula extract, centella asiatica extract, aloe extract and biotin; 0.45%-5% of any one or more of tocopherol (vitamin E), retinyl palmitate and flavor.

4. The skin care lotion according to claim 3, characterized in that: Measured by mass percentage of the total weight of the transdermal composition, the composition of the skin care lotion includes: 5%-20% or 10-20% or 13-18% glycerol; 5%-20% or 5-15% or 8-12% emulsifier; 1%-10% or 3-8% butylene glycol or propylene glycol; 0.5-5% or 1-3% laurocapram; 0.3%-3% or 1%-2% carboxymethyl chitosan; 60%-70% water; Any one or more of 1%-10% yeast extract, 0.3%-3% sodium ascorbyl phosphate, 0.5%-10% superoxide dismutase, 0.005%-1% soluble collagen, 0.005%-1% elastin, 0.005%-1% hydroxypropyl tetrahydropyrantriol, 0.005%-1% carnosine, 0.005%-1% sodium polyglutamate, 0.01%-2% kelp extract, 0.01%-2% witch hazel extract, 0.01%-2% calendula extract, 0.01%-2% centella asiatica extract, 0.01%-2% aloe extract, and 0.01%-2% biotin; Any one or more of 0.3%-3% tocopherol (vitamin E), 0.1%-1% retinyl palmitate and 0.05%-1% flavor.

5. The skin care lotion according to claim 3 or 4, characterized in that: in, The emulsifier is selected from any one or more of polysorbate 80, polysorbate 60, polysorbate 20 and sorbitan oleate.

6. The skin care lotion according to any one of claims 1 to 5, characterized in that: Contains preservatives, Preferably, the skin care lotion contains 0.1-1% by weight of preservatives, based on the weight percentage of the total weight of the skin care lotion. Furthermore, the preservative is selected from any one or more of phenoxyethanol, methylparaben, ethylparaben, propylparaben, benzyl alcohol and sorbic acid.

7. Use of any one of claims 1 to 6 in achieving skin care effects, wherein: When in use, it is applied transdermally by any one or more of the following methods: applying, transdermal instrument, patching and spraying.

8. Use of any one of the skin care emulsions of claims 1 to 6 in the preparation of a medicine or product for achieving skin care effects.

Images