Transdermal compositions and uses thereof

Abstract

The present invention provides a transdermal composition that allows an active substance to effectively penetrate the skin and exert a predetermined effect, and uses thereof, wherein the transdermal composition contains an active substance that can exert a predetermined effect, and the transdermal composition is in the form of an emulsion, in which, calculated in mass percentages based on the total weight of the transdermal composition, the oil phase accounts for 15% to 50%, or 30% to 45%, or 35% to 40%, the emulsifier accounts for 35% to 65%, or 40% to 50%, and the aqueous phase accounts for 10% to 35%, or 10% to 25%.

Description

[Technical Field]

[0001] The present invention is in the field of biotechnology and specifically relates to transdermal compositions and their uses. [Background technology]

[0002] Oral administration is the most commonly used method of drug administration because it is simple, safe, and readily accepted by patients. However, oral administration is subject to low drug solubility and stability, and to gastrointestinal damage and the first-pass effect in the liver after oral administration, resulting in low bioavailability.

[0003] Injection administration causes fluctuations in blood drug concentrations, the drug's half-life is short, and frequent injections can cause pain at the injection site and other side effects, leading to lower patient compliance. Currently, most polymers are administered by injection, making it difficult to continue long-term treatment.

[0004] Transdermal drug delivery systems (TDD) or transdermal therapeutic systems (TTS) are a method of absorbing drugs through the skin. They are a new drug administration route that allows drugs to be absorbed into the human blood circulation through the skin and reach effective blood concentrations, thereby enabling the treatment or prevention of diseases.

[0005] Transdermal drug delivery systems are the third most popular drug delivery system after oral administration and injection, and have the advantages of avoiding the first-pass effect of the liver and drug damage caused by the digestive tract, being convenient to use, with low toxicity and side effects, and being highly safe.

[0006] In addition to treating common skin conditions, transdermal drug delivery is beginning to be used to treat systemic diseases such as hypertension, diabetes, and cancer. This technology is also used in the medical aesthetics and beauty industries, bringing many benefits to patients and beauty enthusiasts.

[0007] Currently, there are drugs on the market that use transdermal drug delivery systems for estradiol, testosterone, etc., and these have been developed mainly for small molecule, lipophilic drugs.

[0008] Regarding polymeric and hydrophilic ingredients, the skin's natural barrier function makes it difficult for the polymeric and hydrophilic ingredients to penetrate the epidermis and dermis of the skin, reach the site of action, and exert their effects. Summary of the Invention [Problem to be solved by the invention]

[0009] The present invention provides a transdermal composition that allows active substances to effectively permeate the skin and exert a desired effect, particularly allowing biopolymers and hydrophilic active substances to effectively permeate the skin and exert a desired effect, and uses thereof. [Means for solving the problem]

[0010] To this end, the present invention provides the following technical solutions:

[0011] The present invention provides a transdermal composition containing an active substance capable of exerting a predetermined effect, the transdermal composition being in the form of an emulsion, wherein the transdermal composition contains, in terms of mass percentage based on the total weight of the transdermal composition: the oil phase accounts for 15% to 50%, or 30% to 45%, or 35% to 40%, Emulsifiers account for 35% to 65% or 40% to 50%. The aqueous phase accounts for 10% to 35%, or 10% to 25%.

[0012] According to the detection, the particle size distribution of the granules in the emulsion is 0.01 μm to 1 μm.

[0013] The transdermal composition provided by the present invention also comprises, in the transdermal composition, calculated as a mass percentage of the total weight of the transdermal composition: the oil phase accounts for 15% to 50%, or 30% to 45%, or 35% to 40%, Emulsifiers account for 35% to 65% or 40% to 50%. The aqueous phase is characterized by occupying 10% to 35%, or 10% to 25%.

[0014] The transdermal composition provided by the present invention is also characterized in that the mass ratio of oil phase:water phase:emulsifier is 2-3:2.4-4:1, or 2-3:2.4-3.7:1.

[0015] The transdermal composition provided by the present invention is characterized in that the emulsion matrix component contains, calculated as a mass percentage in the total weight of the transdermal composition, 1% to 15% oleic acid, 10% to 30% emulsion stabilizer, 5% to 15% any one or more of propylene glycol and butylene glycol, 0.5% to 4% menthol, 1% to 5% laurocapram, 35% to 65% emulsion, and 10% to 20% water and other components.

[0016] The transdermal composition provided by the present invention also includes an emulsifier comprising sorbitan oleate and one or more selected from polysorbate 80, polysorbate 60, and polysorbate 20, calculated as a percentage by mass: The transdermal composition is characterized in that sorbitan oleate accounts for 10% to 15% of the total weight of the transdermal composition, and the sum of any one or more of polysorbate 80, polysorbate 60, and polysorbate 20 accounts for 25% to 50% or 30% to 40% of the total weight of the transdermal composition.

[0017] The transdermal composition provided by the present invention is also characterized in that the trix components of the emulsion, calculated as mass percentages in the total weight of the transdermal composition, include 1% to 15% oleic acid, 10% to 30% emulsion stabilizer, 5% to 15% any one or more of propylene glycol and butylene glycol, 0.5% to 4% menthol, 1% to 5% laurocapram, 35% to 65% emulsion, and 10% to 20% water.

[0018] The transdermal compositions provided by the present invention also contain a preservative, Preferably, the transdermal composition contains 0.1% to 1% by weight of a preservative, calculated as a mass percentage based on the total weight of the composition, and further characterized in that the preservative is selected from any one or more of phenoxyethanol, methylparaben, ethylparaben, propylparaben, benzyl alcohol, and sorbic acid.

[0019] The transdermal compositions provided by the present invention also have the characteristic that the active substance comprises a biomolecular substance, and preferably the biomolecular substance is permeable to biomolecular substances of approximately 300 kDa or less, or approximately 150 kDa or less.

[0020] The transdermal composition provided by the present invention also has the characteristic of being able to achieve a predetermined effect, which may be one or more of a therapeutic effect, a cosmetic effect, and a skin care effect.

[0021] The transdermal composition provided by the present invention also has a structure that allows the biomolecular substance to be delivered to target cells and achieve the function of cleaving exocytosis fusion of the target cells, Furthermore, the biomolecular substance is a clostridial neurotoxin, Furthermore, the clostridial neurotoxin is a type A, type B, type C, type D, type E, type F, or type G botulinum toxin, and / or the light chain of the clostridial neurotoxin comprises an amino acid sequence having at least 35%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% identity to any one of SEQ ID NOs: 1 to 7, and / or the heavy chain of the clostridial neurotoxin comprises an amino acid sequence having at least 35%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% identity to any one of SEQ ID NOs: 8 to 14.

[0022] The transdermal composition provided by the present invention is also characterized in that the active substance contains yeast extract, plant extract, and other substances that can play one or more roles among skin whitening, moisturizing, anti-aging, repair, and cell proliferation, and the active substance is different from the aforementioned matrix component elements (this description is intended to distinguish the main functions of the components and explain the proportions occupied by the components, The matrix component cannot contain elements that have the same function as the active substance. This is not intended to be limiting. That is, the matrix component may of course contain a component that acts as an active substance, but when it is used mainly as a matrix component, the role of the matrix component is emphasized in order to distinguish the ratio.

[0023] The transdermal composition provided by the present invention is also characterized in that the active substance comprises any one or more of yeast extract, sodium ascorbyl phosphate, sea buckthorn seed oil, any vitamin, retinyl palmitate, aloe vera extract, allantoin, licorice extract, Panax ginseng extract, Rheum palmatum extract, Centella asiatica extract, tea leaf extract, and witch hazel extract.

[0024] The transdermal compositions provided by the present invention are characterized in that their ingredients are as shown in Table 6 or Table 7.

[0025] The present invention further provides uses of the transdermal compositions in achieving therapeutic, cosmetic, and skin care benefits.

[0026] The use provided by the present invention is characterized in that, when used, it is applied transdermally by one or more of application, transdermal device, patch, and spray.

[0027] The present invention further provides the use of said transdermal composition in the manufacture of a medicament or product for achieving therapeutic, cosmetic and skin care benefits.

[0028] The present invention provides a method for producing the above transdermal composition, blending and mixing the compatible oil phase raw materials together, and blending and mixing the resulting mixtures together to obtain a final oil phase blend; The method is characterized by including a step of adding the aqueous phase ingredients directly or after dissolving them in the final oil phase mixture and mixing them.

[0029] The present invention also provides a method for producing a transdermal composition, comprising: The components of the transdermal composition are oleic acid, oleyl alcohol, propylene glycol, menthol, laurocapram, sorbitan oleate, any one or more of polysorbate 80, polysorbate 60, and polysorbate 20, water, and an effective amount of a biomolecular substance; The manufacturing method is uniformly mixing a certain amount of polysorbate-80, polysorbate-60, and sorbitan oleate, adding a certain amount of oleyl alcohol and oleic acid and continuously uniformly mixing, and then adding a certain amount of laurocapram and phenoxyethanol and continuously uniformly mixing to obtain a first oil phase group; Mixing and dissolving a fixed amount of propylene glycol and menthol to obtain a second oil phase; a step of uniformly mixing the first oil phase group and the second oil phase group to obtain a final oil phase mixture; The method is characterized by including the steps of adding an effective amount of a biomolecular substance to the final oil phase mixture, mixing uniformly, and supplementing with water.

[0030] The present invention also provides a method for producing a transdermal composition, comprising: The components of the transdermal composition are Oleic acid, oleyl alcohol, propylene glycol, menthol, laurocapram, sorbitan oleate, and one or more of polysorbate 80, polysorbate 60, and polysorbate 20, water, yeast extract, sodium ascorbyl phosphate, sea buckthorn seed oil, various vitamins, retinyl palmitate, aloe vera extract, allantoin, licorice extract, panax ginseng extract, rhubarb extract, centella asiatica extract, tea leaf extract, and witch hazel extract. The manufacturing method is uniformly mixing a predetermined amount of polysorbate-80, polysorbate-60, and sorbitan oleate, adding a predetermined amount of oleyl alcohol, oleic acid, sea buckthorn seed oil, tocopherol, and retinyl palmitate, and then uniformly mixing the resulting mixture; and then adding a predetermined amount of laurocapram and phenoxyethanol, and then uniformly mixing the resulting mixture to obtain a first oil phase group. Mixing and dissolving a fixed amount of propylene glycol and menthol to obtain a second oil phase; a step of uniformly mixing the first oil phase group and the second oil phase group to obtain a final oil phase mixture; The method is characterized by including the step of sequentially adding a fixed amount of plant extract, yeast extract, sodium ascorbyl phosphate, nicotinamide and allantoin dissolved in water to the final oil phase mixture, mixing uniformly, and then supplementing with water. [Effects of the Invention]

[0031] Role and effect of this invention The transdermal composition provided by the present invention is in the form of an emulsion. By appropriately adjusting the ratio of the oil phase, the water phase, and the emulsifier in the emulsion, effective skin penetration can be achieved and the desired effects can be exhibited. In particular, in the case of a polymeric substance and a hydrophilic substance, effective skin penetration can be achieved and the desired effects can be exhibited. [Brief explanation of the drawings]

[0032] [Figure 1] 1 is a transmission electron microscope photograph of particles of the transdermal compositions prepared in Examples 1 to 6. [Figure 2] FIG. 1 is a graph showing the results of an experiment to verify the effectiveness of the transdermal compositions prepared in Examples 1 to 6 in a mouse body. [Figure 3] 1 is a comparison of skin elasticity R2 values ​​before and after use of the transdermal composition prepared in Example 8 (the larger the R2 measurement value, the better the skin elasticity). [Figure 4] 1 is a comparison of the skin firmness F4 values ​​before and after use of the transdermal composition prepared in Example 8 (the higher the F4 measurement value, the better the skin firmness). [Figure 5] 1 is a comparison diagram of the area ratio of wrinkles under the eyes before and after use of the transdermal composition prepared in Example 8 (the smaller the measurement value under the eyes, the smaller the area ratio of wrinkles under the eyes). [Figure 6] 1 is a comparison chart of clinical evaluation-fine wrinkle grades of the face before and after use of the transdermal composition prepared in Example 8 (the smaller the measured value, the smaller the facial fine wrinkle area ratio). [Figure 7] 1 is a comparative chart showing clinical evaluations of nasolabial fold grade, nasolabial fold grade, forehead wrinkle grade, and corner of mouth wrinkle grade before and after use of the transdermal composition prepared in Example 8 (the smaller the measured value, the smaller the fine wrinkle area ratio). [Figure 8] Transdermal testing of the transdermal composition. DETAILED DESCRIPTION OF THE INVENTION

[0033] Specific examples of the present invention will be described below with reference to the drawings. With regard to the specific methods and materials used in the examples, those skilled in the art can make conventional substitutions or selections based on existing technologies in accordance with the technical concept of the present invention, and the present invention is not limited to the specific description of the examples.

[0034] Unless otherwise specified, all methods used in the examples are conventional methods, and all materials, reagents, etc. used are commercially available products unless otherwise specified.

[0035] An emulsion is a liquid preparation containing two immiscible liquid phases, one of which is dispersed in the other in the form of small droplets, forming a heterogeneous dispersion. The droplet-forming phase is called the dispersed, internal, or discontinuous phase, and the other liquid phase is called the dispersion medium, external, or continuous phase.

[0036] Emulsions consist of an oil phase (denoted by O), an emulsifier, and an aqueous phase (denoted by W). Depending on the type, nature, and phase volume ratio (φ) of the emulsifier, oil-in-water (O / W) or water-in-oil (W / O) emulsions or multiple emulsions can be formed.

[0037] Aqueous phase: water or an aqueous solution. An aqueous solution refers to a solution in which water-soluble raw materials are dissolved in water.

[0038] Oil phase: Refers to raw materials that are not easily soluble in water dissolved in an oily substance.

[0039] The transdermal composition provided by the present invention contains an effective amount of an active substance capable of exerting a predetermined effect. The transdermal composition is in the form of an emulsion, and the active substance is incorporated into the matrix of the emulsion.

[0040] The predetermined role is to achieve one or more of a therapeutic effect, a cosmetic effect, and a skin care effect.

[0041] The emulsion matrix referred to in the present invention is also called an auxiliary material component, and is used for the active substance, that is, it is mainly used as a matrix component to form an emulsion.Of course, the emulsion matrix may also contain components that have the same effect as the active substance.For the convenience of explanation, the emulsion matrix is ​​described separately, but the overall components of the emulsion should be considered as a whole, including the emulsion matrix and other components, and the proportions of the oil phase, emulsifier, and aqueous phase must comply with the present invention.

[0042] The limitations for the transdermal compositions of the present invention are as shown in Tables 1 and 2. [Table 1] [Table 2]

[0043] In Tables 1 and 2, the entire transdermal composition is composed of three components: an oil phase, an aqueous phase, and an emulsifier, and contains an "effective amount of an active substance capable of performing a predetermined role." In other words, the active substance may be present in either the oil phase or the aqueous phase.

[0044] The particle size distribution of the granules in the emulsion of the present invention is 0.01 μm to 1 μm, or the emulsion contains granules with a particle size of 100 to 200 nm. Preferably, the emulsion of the present invention is a water-in-oil type.

[0045] Calculated as a mass percentage of the total weight of the transdermal composition, the emulsion matrix is ​​as shown in Table 3. [Table 3]

[0046] Furthermore, calculated as a mass percentage: Sorbitan oleate accounts for 10% to 15% of the total weight of the transdermal composition, and the sum of any one or more of polysorbate 80, polysorbate 60, and polysorbate 20 accounts for 30% to 40% of the total weight of the transdermal composition.

[0047] Further examples of matrix components for transdermal compositions include those shown in Table 4. [Table 4]

[0048] In one example, the transdermal composition further comprises a preservative, preferably 0.1% to 1% by weight of the preservative calculated as a mass percentage based on the total weight of the transdermal composition, and more preferably, the preservative is selected from any one or more of phenoxyethanol, methylparaben, ethylparaben, propylparaben, benzyl alcohol, and sorbic acid.

[0049] The active substance of the present invention includes a biomolecular substance. Preferably, the transdermal composition is permeable to biomolecular active substances having a size of approximately 300 kDa or less, or approximately 150 kDa or less. Here, "approximate" refers not only to the following range, but also to a relatively close but relatively large range, such as 305 kDa. Preferably, the range is greater than 50 kDa, i.e., 350 kDa or less, or 200 kDa or less.

[0050] Da is short for Dalton, a common unit of molecular weight, which is the algebraic sum of the atomic weights of all atoms in a molecule. The Dalton (Da) is numerically equivalent to the relative molecular weight.

[0051] In biochemistry, molecular biology, and proteomics, the unit kDa (kilodalton) is often used to describe biological macromolecules such as proteins. 1 kDa refers to a molecule with a relative molecular weight of 1000, whereas macromolecules refer to biological substances with a relative molecular weight of 5000 or more, or even over 1 million, such as peptides, proteins, nucleic acids, polysaccharides, and antibodies.

[0052] In one example, the biomolecular substance has a structure that can be delivered to a target cell and perform the function of severing the exocytosis fusion of the target cell.

[0053] Specifically, non-cytotoxic proteases exert their effects by proteolytically cleaving intracellular transport proteins called SNARE proteins (see Gerald K (2002) - Cell and Molecular Biology (4th edition) John Wiley & Sons, Inc.).

[0054] The acronym SNARE is derived from the term Soluble NSF Attachment Receptor, where NSF stands for N-ethylmaleimide-Sensitive Factor.

[0055] SNARE proteins are required for the fusion of intracellular vesicles and thus for the secretion of molecules from cells via vesicular transport. Non-cytotoxic proteases function as zinc-dependent endopeptidases and exhibit high substrate specificity for SNARE proteins. SNARE proteins bind to the membrane of secretory vesicles or the plasma membrane, mediating the fusion of secretory vesicles with the plasma membrane and promoting exocytosis of molecules, thereby allowing the contents of the vesicle to be expelled from the cell. Targeting and cleaving neuronal SNAREs inhibits neurotransmitter release and causes paralysis. Therefore, when non-cytotoxic proteases are delivered to the desired target cells, they can inhibit cellular secretion from the desired target cells.

[0056] Three proteins at the synaptic terminal, synaptobrevin (VAMP) on the synaptic vesicle membrane, syntaxin (SNP) on the presynaptic membrane, and SNAP-25, are all called SNARE proteins.

[0057] In one example, the cell is a neuron and the exocytosis fusion apparatus protein is a neuronal SNARE protein. Cleaving the SNARE protein inhibits the physiological function of cholinergic nerve conduction, for example, by inhibiting the release of the acetylcholine neurotransmitter.

[0058] Additionally, the biomolecular agent is a clostridial neurotoxin.

[0059] Clostridial toxins can be used for both therapeutic and cosmetic purposes. For example, botulinum toxin can specifically bind to presynaptic membrane receptors on peripheral cholinergic nerve terminals, cleaving the synapse-associated protein SNAP-25, inhibiting the exocytosis of presynaptic membrane vesicles, and inhibiting the release of acetylcholine (Ach) from nerve terminals, resulting in muscle chemical denervation, thereby relaxing muscles and relieving muscle spasms, exerting its therapeutic effects. The muscle chemical denervation effect of botulinum toxin is the rationale for the regulation of muscle hypertonia, movement disorders, and dynamic wrinkles. Furthermore, because multiple injections into targeted muscles can cause muscle atrophy and loss of muscle mass due to disuse, botulinum toxin is also used in body shaping. Botulinum toxin is currently used to treat disorders such as hemifacial spasm, essential blepharospasm, spasmodic torticollis, spastic cerebral palsy, post-stroke limb spasms, tremors, and other types of dystonia. Applications of botulinum toxin in minimally invasive plastic surgery include the removal of various dynamic wrinkles such as glabellar lines, forehead wrinkles, and crow's feet, as well as facial contouring such as brow height adjustment, mentalis muscle relaxation, jaw lift, and cervical cord injection, and body contouring such as masseter muscle reduction, gastrocnemius muscle reduction, and trapezius muscle reduction.

[0060] As used herein, a clostridial toxin refers to, for example, a naturally occurring clostridial toxin, or a toxin having a structure and function similar to a naturally occurring clostridial toxin.

[0061] Although the various types of toxins differ in their amino acid sequences and immunological properties, they all display similar molecular structures. Clostridial neurotoxins are produced by toxigenic clostridia as single-chain polypeptides of approximately 150 kD. They are activated only after cleavage by bacterial or in vitro proteases into a dichain form consisting of a light chain (L chain, amino terminal of the toxin, 50 kD) and a heavy chain (H chain, carboxy terminal of the toxin, 100 kD) connected by disulfide bonds. The heavy chain consists of two domains: Hn (amino terminal, 50 kD) and Hc (carboxy terminal, 50 kD).

[0062] Furthermore, the clostridial neurotoxin is a botulinum toxin type A, B, C, D, E, F, or G, and / or the light chain of the clostridial neurotoxin comprises an amino acid sequence having at least 35%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% identity to any one of SEQ ID NOs: 1-7, and / or the heavy chain of the clostridial neurotoxin comprises an amino acid sequence having at least 35%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% identity to any one of SEQ ID NOs: 8-14. The sequences are as shown in Table 5.

[0063] [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4]

[0064] Botulinum toxin-related products have broad potential uses in the treatment of clinical spastic disorders, excessive glandular secretions, neuropathic pain, and cosmetic wrinkle removal, etc. However, due to the large size of these products, which are currently administered by injection, it is difficult for them to penetrate the skin directly to reach the site of action.

[0065] The transdermal composition of the present specification was verified through an in vivo efficacy experiment in mice to demonstrate that the biological polymer can penetrate the skin and exert its intended effect.

[0066] Preferably, the effective amount of clostridial toxin comprises no more than 0.02% of the total transdermal composition, more preferably no more than 0.015%, and even more preferably no more than 0.012%.

[0067] In one example, the active substances include any one or more of 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 proliferation. It is also stipulated that the active substance is different from the aforementioned matrix component elements in order to distinguish it from substances that may have the same effect as the active substance in the matrix component. (This explanation is to distinguish the main functions of the ingredients and explain the proportions of the ingredients. The matrix component cannot contain a component that has the same function as the active substance. This is not intended to be limiting. That is, the matrix component may of course contain a component that acts as an active substance, but when it is used primarily as a matrix component, the role of the matrix component is emphasized to distinguish the proportion. For example, when glycerol is the main component of the matrix, glycerin also has the same effect as an active substance (moisturizing and lubricating the skin), but for the convenience of explanation, glycerol is used primarily as a component of the matrix component, and even when glycerol also plays the role of an active substance, it is no longer classified as an active substance; this is classified as a formal component for the convenience of explanation protection.

[0068] Furthermore, in specific embodiments, the active substance includes any one or more of yeast extract, sodium ascorbyl phosphate, sea buckthorn seed oil, any vitamin (e.g., vitamin E, nicotinamide), retinyl palmitate, aloe vera extract, allantoin, licorice extract, Panax ginseng extract, Rheum palmatum extract, Centella asiatica extract, tea leaf extract, and witch hazel extract. Of course, the active substance referred to in this paragraph may also include the aforementioned biomolecular substances.

[0069] Vitamins are essential organic substances for the human body. The addition of vitamins and their derivatives has whitening and anti-aging effects. Vitamin A promotes epidermal cell proliferation and increases the formation of collagen and elastin in the dermis. Vitamin E is a natural antioxidant that can accelerate metabolism and improve skin elasticity. Vitamin C has a strong antioxidant effect, promotes collagen synthesis, and inhibits collagen degradation. Vitamin C and vitamin E exert a synergistic effect in scavenging free radicals. Nicotinamide (vitamin B3) reduces wrinkles by increasing collagen. Nicotinamide also enhances histone acetyltransferase activity in human fibroblasts and simultaneously reduces the content of histone acetyltransferase and histone H4 in senescent cells.

[0070] In addition to vitamins essential for the human body, natural plant active ingredients such as tea extract, ginseng extract, Centella asiatica extract, and licorice extract have also been proven to have anti-wrinkle, firming, and whitening effects, etc. These raw materials are currently widely used in anti-wrinkle, firming, and whitening cosmetics.

[0071] However, in conventional cosmetics, the active ingredients contained in these raw materials are only absorbed to the skin surface, resulting in limited effectiveness.The transdermal composition of the present invention can significantly enhance the transdermal absorption of active ingredients, and its actual effectiveness has been verified in human efficacy experiments.

[0072] Tables 6 and 7 are two examples of ingredients for transdermal compositions provided by the present invention. [Table 6] [Table 7]

[0073] The transdermal composition of the present invention is used to achieve therapeutic, cosmetic, and skin care effects. When used, it can be administered by one or more of the following methods: application, transdermal device, patch, and spray. When used, it can be administered by one or more of the following methods: application, transdermal device, patch, and spray.

[0074] The transdermal compositions of the present invention can also be used in the manufacture of medicines or products to achieve therapeutic, cosmetic, and skin care benefits.

[0075] The present invention further comprises: miscible oil phase raw materials are respectively miscible and blended, and the resulting mixtures are respectively miscible and blended to obtain a final oil phase mixture; adding the aqueous phase ingredients directly or after dissolving them in the final oil phase mixture and mixing.

[0076] In one example, a transdermal composition is prepared containing one or more of the following ingredients: polysorbate-80, polysorbate-60, and sorbitan oleate, water, and an effective amount of a biomolecular substance. uniformly mixing the determined amounts of polysorbate-80, polysorbate-60, and sorbitan oleate, then adding the determined amounts of oleyl alcohol and oleic acid and uniformly mixing, then adding the determined amounts of laurocapram and phenoxyethanol and uniformly mixing to obtain a first oil phase composition; Mixing and thoroughly dissolving a predetermined amount of propylene glycol and menthol to obtain a second oil phase composition; The method includes the steps of uniformly mixing the first oil phase composition and the second oil phase composition to obtain a final oil phase mixture, adding an effective amount of a biomolecule to the final oil phase mixture and mixing uniformly, and adding water.

[0077] In one example, a transdermal composition is prepared containing the following ingredients: oleic acid, oleyl alcohol, propylene glycol, menthol, laurocapram, sorbitan oleate, and any one or more of polysorbate 80, polysorbate 60, and polysorbate 20, water, and an effective amount of yeast extract, sodium ascorbyl phosphate, sea buckthorn seed oil, any vitamins, retinyl palmitate, aloe vera extract, allantoin, licorice extract, Panax ginseng extract, Rheum palmatum extract, Centella asiatica extract, tea leaf extract, and witch hazel. Specifically, the preparation method at this time is as follows: uniformly mixing the determined amounts of polysorbate-80, polysorbate-60, and sorbitan oleate, then adding the determined amounts of oleyl alcohol, oleic acid, sea buckthorn seed oil, tocopherol, and retinyl palmitate and uniformly mixing them, and then adding the determined amounts of laurocapram and phenoxyethanol and uniformly mixing them to obtain a first oil phase composition; Mixing and thoroughly dissolving a predetermined amount of propylene glycol and menthol to obtain a second oil phase composition; uniformly mixing the first oil phase composition and the second oil phase composition to obtain a final oil phase mixture; The method includes the step of sequentially adding a fixed amount of plant extract, yeast extract, and sodium ascorbyl phosphate, nicotinamide, and allantoin, each dissolved in water, to the final oil phase mixture and mixing uniformly, and then replenishing with water.

[0078] In the present invention, the water is preferably any one or a combination of a plurality of distilled water, drinking water, sterilized water, ultrapure water, and deionized water. [Example]

[0079] Examples 1 to 6 In Examples 1 to 6, transdermal compositions were prepared using botulinum toxin type A as the active substance. Specific ingredients are as shown in Table 8. [Table 8]

[0080] Manufacturing method: Step 1: According to the formulation, a fixed amount of any one or more of polysorbate-80, polysorbate-60, and polysorbate 20 and sorbitan oleate was added to a container and stirred for 5 minutes, a fixed amount of oleyl alcohol and oleic acid were successively added to the container and stirred for 5 minutes, and a fixed amount of laurocapram and phenoxyethanol were successively added to the container and stirred uniformly at room temperature (rotation speed: 200-250 rpm, 5 minutes). Step 2: According to the formula, propylene glycol and menthol were weighed and mixed separately, heated to 60°C (for about 10 minutes) to completely dissolve them, and then added to the solution obtained in Step 1 and stirred uniformly at room temperature (rotation speed: 200-250 rpm, for 5 minutes). Step 3: 10 mg of botulinum toxin type A was added to the solution prepared in step 2, and the mixture was stirred at room temperature to mix evenly (rotation speed: 200-250 rpm, 5 minutes). Step 4: The solution prepared in step 3 was supplemented with purified water and stirred at room temperature to mix uniformly.

[0081] Example 7 This experiment detected transdermal efficiency. The botulinum toxin type A added in Example 1 was replaced with BSA protein, and the BSA protein emulsion produced had a concentration of 0.62 mg / ml, and a transdermal test was carried out using the Franz diffusion cell method.

[0082] Test Equipment and Materials 1. Reagents Phosphate buffer solution, protein standards, test products, protein detection kits (ultra-sensitive). 2.Consumables For the transdermal evaluation, isolated pig skin, centrifuge tubes, EP tubes, pipette tips, and precoated microtiter plates were used. 3. Equipment TP-6 transdermal diffusion device (Tianjin Jingtuo Instrument Technology Co., Ltd.), multifunction microplate reader (Thermo Scientific, Varioskan Lux), medical refrigerator (Haier Biomedical), biochemical incubator (SHANGHAI BLUEPARD INSTRUMENTS CO., LTD), pipette (Eppendorf AG).

[0083] 1. Preparation before the experiment: (1) The pig skin required for the experiment was removed from a refrigerator at -20°C, thawed at 4°C, and then washed with saline. (2) Before the experiment, the supply chamber and the diffusion chamber (receiving chamber) were cleaned and received the liquid and the stirring bar from the supply chamber. (3) Pure water was poured into the water tank until it reached the same height as the diffusion cell frame, and the temperature was set at 32°C and the rotation speed was set. (4) Once the temperature of the TP-6 ​​transdermal diffusion device reached the set temperature, the pig skin was placed in the supply chamber and stirred at a uniform constant speed between the supply chamber and the diffusion chamber to maintain an isothermal state and a uniform exudate concentration in the solution.

[0084] 2. Experimental Method: (1) Two experimental groups and one blank group were set up. (2) Addition of BSA protein test solution: 150 μl (0.62 mg / ml) of the BSA protein emulsion test solution prepared in Example 1 and 150 μl (1 mg / ml) of the BSA protein test solution dissolved in PBS were added to two supply chambers, respectively. 3 ml of PBS was added to the blank group. (3) The receiving chamber was filled with PBS, and four 1.5 ml EP tubes were prepared. Every hour, 400 μl of the liquid in the receiving chamber was sampled through the sampling port of the receiving chamber, followed by OD detection and labeling with the sampling time and experimental group number. The total sampling time was 6 hours. After sampling, each receiving chamber needed to be refilled with 400 μl of PBS to prevent the volume of the receiving chamber from changing. All collected samples were stored in a refrigerator at 4°C for later use.

[0085] 3.ELISA experiment detection: 1) Preparation of reagents: a. Before use, allow all reagents to warm to room temperature (18-25°C). According to the microplate reader instructions, set the detection wavelength to 450 nm and preheat the plate for 15 minutes before reading. b. Wash Buffer: 20 ml of concentrated wash buffer was diluted with 580 ml of deionized water to make 600 ml of wash buffer. c. Standard working solution: First, the standard was centrifuged at 1000 g for 1 minute, and 2 ml of standard dilution solution was added and mixed uniformly. 300 μl of the 20,000 ng / ml standard dilution solution was pipetted into the first tube and mixed uniformly to prepare a 6,666.7 ng / ml working solution. Next, 300 μl of the solution was transferred from the front tube to the back tube. Each tube was mixed uniformly before the next transfer. Six standard dilutions were set up, each at 20,000 ng / ml, 6,666.7 ng / ml, 2,222.2 ng / ml, 740.7 ng / ml, 246.9 ng / ml, and 0 ng / ml. d. Detection Reagent A Working Solution: Calculate the required volume (100 μl / well) before the experiment. Prepare 100-200 μl more than the calculated volume. Before use, briefly centrifuge the stock solution tube and dilute 100x Concentrated Detection Reagent A with Detection Diluent A to make 1x Working Solution A (e.g., 10 μl of Detection Reagent A + 990 μl of Detection Reagent A Diluent). e. Detection Reagent B Working Solution: Calculate the required amount (100 μl / well) before the experiment. When preparing, prepare 100-200 μl more than the calculated amount. Before use, briefly centrifuge the stock solution tube and dilute 100x Concentrated Detection Reagent B with Detection Diluent B to make 1x Working Solution B (e.g., 10 μl of Detection Reagent A + 990 μl of Detection Reagent B diluent).

[0086] 2) Experimental method (1) Standard wells, sample wells, and blank wells were placed in each well. Six different concentrations of standard samples (50 μL per well, including the zero well) were added in turn, and 50 μL of test sample (sample collected from the sampling port of the receiving chamber) was added to the other wells. Immediately after that, 50 μL of detection reagent A working solution was added in each well. The wells were gently shaken to mix evenly, covered with the sealing film provided with the kit, and incubated at 37°C for 60 minutes. (2) Discard the liquid from all wells, add 350 μL of wash buffer to each well, and after immersion for 60 seconds, pour off the liquid from each well and pat dry on clean absorbent paper. This washing step was repeated a total of three times. (3) 100 μL of diluted detection reagent B solution was added to each well, covered with sealing film, and incubated at 37°C for 30 minutes. (4) The liquid in each well was discarded, and the washing process in step 2 was repeated five times. (5) 90 μL of TMB reagent was added to each well, covered with a new sealing film, and incubated in the dark at 37°C for 10 to 20 minutes. (6) Add 50 μL of the reaction stop solution to each well in the same order as the coloring solution. Shake gently to mix evenly. (7) After making sure that there were no air bubbles or water droplets at the bottom of the wells of the ELISA plate, the absorbance OD value of each well at 450 nm was immediately measured and the ELISA reading was recorded.

[0087] 4. Test Data and Conclusions (1) The standard curve obtained by fitting the BSA standard according to the four parameters is shown in Figure 1. Standard curve fitting: Processing of OD value data of detected standard proteins: The data was fitted with four parameters to obtain a standard curve. The four parameter fitting equation is as follows:

number

[0088] (2) Sample detection results Table 9 shows the BSA concentrations that penetrated into the skin of various test samples at different time periods, calculated based on the standard curve. [Table 9]

[0089] Based on the results, the skin penetration was calculated. (1) In Comparative Example 2, a BSA emulsion protein test solution was prepared. After 24 hours, the concentration in the receiving chamber was 147.1 ng / ml and its volume was 15 ml, while the BSA concentration in the supply chamber was 0.62 mg / ml and the volume added to the supply chamber was 150 μl. Based on this, the transdermal absorption efficiency of the BSA emulsion prepared in Example 5 was calculated to be 2.37%. (2) A BSA protein test solution was prepared in PBS. After 24 hours, the concentration in the receiving chamber was 125.4 ng / ml, with a volume of 15 ml, while the BSA concentration in the supply chamber was 1 mg / ml. The volume added to the supply chamber was 150 μl. Based on this, the transdermal absorption efficiency of BSA was calculated to be 1.25%.

[0090] This shows that the transdermal effect of Example 1 is very good compared to the control group.

[0091] Example 8 The transdermal compositions prepared in Examples 1 to 6 were subjected to efficacy verification experiments in mice.

[0092] SPF CD-1 (ICR) mice weighing 17-19 g were used. After anesthetizing with tribromoethanol, the skin on the right hind leg was shaved. Before each application, the application site was cleaned with a medical cotton swab soaked in 10% alcohol, allowed to dry, and then the drug was applied. 200 μl of the prepared transdermal composition was applied to the skin of the right hind leg of each mouse once daily for five consecutive days. As a control, no transdermal composition was applied to the left hind leg of the mice.

[0093] The experimental results showed that 3 to 5 days after application, the right hind paws of the mice showed a closed toe phenotype (two or three closed toes). The closed toe phenotype is shown in Figure 2, and this phenotype was not observed in the control group.

[0094] According to a literature report ("An Alternative In Vivo Method to Refine the Mouse Bioassay for Botulinum Toxin Detection", Temeri D Wilder-Kofie et al., Vol. 61, No. 3 June 2011), injection of botulinum toxin type A into the gastrocnemius muscle of mice resulted in a toe-closure phenotype. When the transdermal composition of the present specification was applied to mouse skin, it also induced a toe-closure phenotype after injection of botulinum toxin type A. This indicates that the transdermal composition of the present specification achieves effective transdermal penetration of botulinum toxin type A. In other words, it was found that the transdermal composition of the present specification allows a polymer to effectively penetrate the skin and exert its intended effect.

[0095] Example 9 In this example, a transdermal composition was prepared according to the formulation shown in Table 10. [Table 10]

[0096] Manufacturing method: Step 1: According to the recipe, quantitative amounts of polysorbate-80, polysorbate-20, and sorbitan oleate were added to a reaction vessel and stirred for 5 minutes. Subsequently, quantitative amounts of oleyl alcohol, oleic acid, sea buckthorn seed oil, tocopherol, and retinyl palmitate were added to the reaction vessel and stirred for 5 minutes. Subsequently, quantitative amounts of laurocapram and phenoxyethanol were added to the reaction vessel and mixed uniformly at room temperature (rotation speed: 200-250 rpm, 5 minutes). Step 2: According to the recipe, propylene glycol and menthol were weighed, mixed, and heated to 60°C (approximately 10 minutes) to completely dissolve them. The mixture was then added to the solution obtained in Step 1 and stirred uniformly at room temperature (rotation speed: 200-250 rpm, 5 minutes). Step 3: According to the recipe, a fixed amount of plant extracts (tea leaf extract, ginseng, Centella asiatica extract, aloe vera extract, licorice root extract, Rheum palmatum extract, witch hazel extract, and yeast extract) was added to the solution prepared in Step 2 and stirred at room temperature (rotation speed: 200-250 rpm, 5 minutes). Step 4: According to the recipe, a fixed amount of sodium ascorbyl phosphate was dissolved in water, added to the solution obtained in Step 3, and stirred uniformly at room temperature (rotation speed: 200-250 rpm). Step 5: According to the recipe, a fixed amount of nicotinamide was added and dissolved in water, and the mixture was heated to 60°C until completely dissolved. The mixture was then added to the solution obtained in Step 4 and stirred uniformly at room temperature (rotation speed: 200-250 rpm). Step 6: According to the prescription, a fixed amount of allantoin was dissolved in water, heated to 80°C to completely dissolve it, and then added to the solution obtained in Step 5, followed by stirring uniformly at room temperature (rotation speed: 200-250 rpm). Step 7: Purified water was added to the solution prepared in Step 6 according to the recipe, and the mixture was mixed by stirring at room temperature to the specified volume.

[0097] Example 10 The effectiveness of the transdermal composition prepared in Example 9 was detected in vivo in humans.

[0098] A total of 33 subjects were recruited according to the specified subject criteria, and all 33 subjects ultimately completed the study. The age range was 35 to 55 years old, with 4 men and 29 women, and the average age was 45.27 years (see Table 10 for details).

[0099] Subject selection criteria: 1) Healthy men and women aged 30-55 who have noticeable crow's feet, nasolabial folds, or forehead wrinkles. 2) People who can cooperate well with the experimenters and maintain a regular lifestyle during the study period. 3) A person who can read and understand the entire contents of the informed consent document and voluntarily sign the informed consent document. 4) Humans who agree not to use cosmetics, medicines, or health products that may affect the results during the study period. 5) Other relevant standards.

[0100] Subject Exclusion Criteria: 1) People who have used antihistamines within the past week or tried immunosuppressants within the past month. 2) People who have used anti-inflammatory drugs on the test area within the past two months. 3) People who have had skin diseases (e.g., psoriasis, eczema, cattle lice, skin cancer). 4) Patients with insulin-dependent diabetes mellitus. 5) Patients receiving treatment for asthma or other chronic respiratory diseases. 6) People who have received anti-cancer chemotherapy within the past 6 months. 7) Patients with immunodeficiency or autoimmune disease. 8) Women who are breastfeeding or pregnant. 9) People who have undergone bilateral mastectomy and bilateral axillary lymph node dissection. 10) People who are judged to have scars, pigmentation, atrophy, port wine staining, uneven skin tone, folliculitis, or other damage to the skin being tested that may affect the test results. 11) People with other health problems or chronic illnesses. 12) People who have used topical retinoid A, alpha hydroxy acid, salicylic acid, or hydroquinone within the past 3 months, or who have used prescription drugs (antibiotics, retinoid A, alpha hydroxy acid, steroids) or oral contraceptives within the past 6 months (if they have been taking the same contraceptives in the past 6 months, they can continue to take them). 13) People who are considered by experts or professionals to have other iatrogenic reasons that may affect the evaluation results.

[0101] [Table 11]

[0102] 1.How to use Before going to bed every night, thoroughly wash your face (e.g., with soap and makeup remover). For the product to penetrate more effectively, we recommend wiping it off with a cleansing sheet to thoroughly remove oil. Once your skin is dry, look in the mirror and identify the areas that need wrinkle removal. Open the sample cap and press the bottom of the bottle until the liquid is squeezed out around the roller. Roll the roller back and forth five times until the product is evenly distributed. After application, allow the product to gradually absorb into the skin without wiping it off.

[0103] 2. Frequency of use: Once every other day.

[0104] 3. Purpose of the test The test samples are evaluated for their moisturizing, repairing, anti-wrinkle and firming effects.

[0105] 4. Testing Environment Temperature: 21±1℃ Humidity: 50±10%RH

[0106] 5. Test Method 5.1 Method Reference 1) QB / T 4256-2011 Guidelines for evaluating the moisturizing effect of cosmetics. 2) T / ZHCA 003-2018 Test method for the effect of cosmetics on transepidermal water loss. 3) T / ZHCA 006-2019 Test method for anti-wrinkle efficacy of cosmetics. 4) T / TDCA 003-2021 Test method for the firming efficacy of cosmetics. 5) Safety Technical Standards for Cosmetics (2015 Edition).

[0107] 5.2 Testing Process D0 (refers to the subject's first day of visit): 1) Subject information was registered, and eligible subjects with obvious wrinkles (crow's feet grade ≥ 2, nasolabial fold grade ≥ 1, forehead wrinkle grade ≥ 1) were screened by laboratory technicians. After subjects met the inclusion and exclusion criteria, they signed informed consent. 2) The subjects washed their faces with the designated cleansing samples and sat in a laboratory at 21±1°C and 50±10% RH for 30 minutes. 3) Testing the subjects' skin baseline values: The moisture content of the stratum corneum was measured using a Corneometer, the transepidermal water loss rate was measured using a Tewameter, and the skin's elasticity R2 and firmness F4 values ​​were measured using a Cutometer.Facial fine lines, nasolabial folds, sigmoid colon, forehead wrinkles, and wrinkles at the corners of the mouth were also evaluated by an expert.

[0108] D14 (refers to the subject's visit date 14 days after using the sample): 1) The subjects washed their faces with the designated cleansing samples and sat in a laboratory at 21±1°C and 50±10% RH for 30 minutes. 2) Testing the baseline values ​​of the subjects' skin on day 14: The moisture content of the stratum corneum was measured using a Corneometer, the transepidermal water loss rate was measured using a Tewameter, and the skin's elasticity R2 value and firmness F4 value were measured using a Cutometer. In addition, experts evaluated the fine lines, nasolabial folds, wrinkles on the forehead, and wrinkles at the corners of the mouth on the face. 3) The subjects answered a questionnaire.

[0109] D28 (referring to the subject's visit date 28 days after using the sample): 1) The subjects washed their faces with the designated cleansing samples and sat in a laboratory at 21±1°C and 50±10% RH for 30 minutes. 2) Testing the baseline values ​​of the subjects' skin on day 28: The moisture content of the stratum corneum was measured using a Corneometer, the transepidermal water loss rate was measured using a Tewameter, and the skin's elasticity R2 and firmness F4 values ​​were measured using a Cutometer. In addition, experts evaluated the fine lines, nasolabial folds, wrinkles on the forehead, and wrinkles at the corners of the mouth on the face. Subjects completed a questionnaire and usage records were collected.

[0110] 5.3 Test items (Table 12) [Table 12]

[0111] 6. Statistical methods for data 6.1 Descriptive Statistics Instrumental measurements: mean, standard error, maximum, minimum. Subject self-assessment: percentage of recognition.

[0112] 6.2 Difference analysis Statistical analysis of the data was performed using SPSS statistical analysis software. The measurement data was expressed as mean ± standard error, and a normal distribution test was performed. If the normal distribution requirement was met, a paired t-test was used to compare before and after the experiment; otherwise, a rank sum test of two related samples was used. The rank sum test of two related samples was used to compare the stage-specific data before and after the experiment, and an independent sample t-test or rank sum test was used to compare the test product and the control group. All of the above statistical analyses were two-sided, with a significance level of α = 0.05.

[0113] 7. Test Results Subject completion status A total of 33 subjects were included in this study, and 33 ultimately completed the study. The age range was 35-55 years old, with 4 men and 29 women, and the average age was 45.27 years. Specific information is shown in Table 10. [Table 13]

[0114] 7.2 Feedback regarding adverse reactions during use [Table 14] Note: Please refer to the 2015 edition of the "Cosmetics Safety Technical Standards" for the grading criteria for adverse skin reactions in human testing.

[0115] Result explanation: After using the sample for 14 days, all skin reactions in the observation area were grade 0, and no adverse skin reactions occurred. After using the sample for 28 days, all skin reactions in the observation area were grade 0, and no adverse skin reactions occurred.

[0116] 7.3 Descriptive and statistical analysis of test results for stratum corneum moisture content (higher values ​​indicate higher moisture content in the stratum corneum of the skin) [Table 15] Note: Improvement rate (%) = (test value after using the sample for N days - test value before use) / test value before use × 100% [Table 16]

[0117] Result explanation: Compared to before use, after 14 days of using the sample, the improvement rate of the moisture content of the stratum corneum was 21.23%, which was a significant difference. Compared to before use, after 28 days of using the sample, the improvement rate of the moisture content of the stratum corneum was 41.38%, which was a significant difference.

[0118] 7.4 Descriptive and statistical analysis of transepidermal water loss test results (smaller values ​​indicate a lower transepidermal water loss per unit time and cross-sectional area, and a better skin barrier) [Table 17] Note: Improvement rate (%) = (test value before use - test value after using the sample for N days) / test value before use × 100% [Table 18]

[0119] Result explanation: Compared to before use, the improvement rate of transepidermal water loss after 14 days of sample use was 15.84%, which was a significant difference. Compared to before use, the improvement rate of transepidermal water loss after 28 days of sample use was 28.73%, which was a significant difference.

[0120] 7.5 Skin elasticity R 2 Descriptive and statistical analysis of the value test results (the higher the measured value, the better the skin elasticity) [Table 19] Note: Improvement rate (%) = (test value after using the sample for N days - test value before use) / test value before use × 100% [Table 20]

[0121] FIG. 3 shows the skin elasticity R before and after use of the transdermal composition prepared in Example 9. 2 The comparison of values ​​(R 2 (The higher the measured value, the better the skin elasticity.) Significant differences are indicated: "ns" indicates no significant difference (P≧0.05), "*" indicates significant difference (0.01≦P<0.05), "**" indicates 0.001≦P<0.01, and "***" indicates P<0.001.

[0122] The results showed that the skin elasticity improved 14 and 28 days after using the sample compared to before use. 2 The values ​​were significantly improved (P<0.05), with improvement rates of 11.02% and 20.13%, respectively.

[0123] 7.6 Descriptive and statistical analysis of skin firmness F4 value test results (the lower the measurement value, the better the skin firmness) [Table 21] Note: Improvement rate (%) = (test value before use - test value after using the sample for N days) / test value before use × 100% [Table 22]

[0124] Figure 4 shows a comparison of the skin firmness F4 values ​​before and after use of the transdermal composition prepared in Example 9 (the smaller the F4 measurement value, the better the skin elasticity). Significant differences are indicated by "ns" (P≧0.05), "*" (0.01≦P<0.05), "**" (0.001≦P<0.01), and "***" (P<0.001).

[0125] The results showed that compared with before use, the skin firmness F4 value significantly decreased after 14 and 28 days of sample use (P<0.05), with improvement rates of 8.45% and 18.35%, respectively.

[0126] 7.7 Descriptive and statistical analysis of the test results for the area ratio of under-eye wrinkles (the smaller the measurement value, the smaller the area ratio of under-eye wrinkles) [Table 23] Note: Improvement rate (%) = (test value before use - test value after using the sample for N days) / 100% test value before use [Table 24]

[0127] Figure 5 shows a comparison of the area ratio of under-eye wrinkles before and after use of the transdermal composition prepared in Example 9 (the smaller the measurement value under the eyes, the smaller the area ratio of under-eye wrinkles). Significant differences are indicated: "ns" indicates no significant difference (P≧0.05); "*" indicates significant difference (0.01≦P<0.05); "**" indicates 0.001≦P<0.01; and "***" indicates P<0.001.

[0128] The results showed that the area of ​​under-eye wrinkles was significantly reduced after 28 days of sample use compared with before use (P<0.05), with an improvement rate of 8.46%.

[0129] 7.8 Clinical evaluation - Descriptive and statistical analysis of the test results for the level of facial fine lines and other items (the smaller the measurement, the fewer the number of facial fine lines) FIG. 6 is a comparative diagram of clinical evaluation of facial fine wrinkle levels before and after use of the transdermal composition prepared in Example 9 (the smaller the measured value, the smaller the facial fine wrinkle area ratio). FIG. 7 is a comparative diagram showing clinical evaluations of the level of nasolabial folds, the level of striated lines, the level of wrinkles on the forehead, and the level of wrinkles at the corners of the mouth before and after use of the transdermal composition prepared in Example 9 (the smaller the measured value, the smaller the area ratio of fine wrinkles).

[0130] result: Compared to before use, the improvement rate of facial fine wrinkles in clinical evaluation after 28 days of sample use was 9.09%. Compared to before use, the improvement rate of the clinical evaluation - nasolabial fold level after 14 days of sample use was 11.00%, which was a significant difference. Compared to before use, the improvement rate of the clinical evaluation - nasolabial fold level after 28 days of sample use was 16.00%, which was a significant difference. Compared to before use, the improvement rate of the clinical evaluation - Kawajimon level after 28 days of sample use was 4.82%, which was a significant difference. Compared to before use, the clinical evaluation of the improvement rate of forehead wrinkles after 14 days of sample use was 7.27%, which was a significant difference. Compared to before use, the clinical evaluation of the improvement rate of forehead wrinkles after 28 days of sample use was 12.73%, which was a significant difference. Compared to before use, the clinical evaluation of the improvement rate of wrinkles at the corners of the mouth after 14 days of sample use was 11.63%, which was a significant difference. Compared to before use, the clinical evaluation of the improvement rate of wrinkles at the corners of the mouth after 28 days of sample use was 15.12%, which was a significant difference.

[0131] 8. Test Results Thirty-three subjects used the test sample for 28 consecutive days and the results showed the following effectiveness:

[0132] ●Equipment test results: 1) Compared to before use, the moisture content of the stratum corneum increased significantly after 14 and 28 days of sample use (P<0.05), with improvement rates of 21.23% and 41.38%, respectively. 2) Compared to before use, the transepidermal water loss rate of the skin was significantly reduced after 14 and 28 days of sample use (P<0.05), with improvement rates of 15.84% and 28.73%, respectively. 3) Compared to before use, the skin elasticity R2 values ​​were significantly improved after 14 and 28 days of sample use (P<0.05), with improvement rates of 11.02% and 20.13%, respectively. 4) Compared to before use, the skin firmness F4 values ​​were significantly improved after 14 and 28 days of sample use (P<0.05), with improvement rates of 8.45% and 18.35%, respectively. 5) Compared to before use, the area of ​​wrinkles under the eyes on the skin was significantly reduced after 28 days of sample use (P<0.05), with an improvement rate of 8.46%.

[0133] Visual evaluation results: 1) After 28 days of sample use, the level of fine wrinkles on the face was significantly reduced in clinical evaluation compared to before use (P<0.05), with an improvement rate of 9.09%. 2) Compared to before use, the level of nasolabial folds in clinical evaluation was significantly reduced 14 and 28 days after use of the sample (P<0.05), with improvement rates of 11.00% and 16.00%, respectively. 3) Compared with the level before use, the level of Kawajimon in clinical evaluation was significantly reduced after 28 days of sample use (P<0.05), with an improvement rate of 4.82%. 4) Compared to before use, the level of wrinkles on the forehead in clinical evaluation was significantly reduced 14 and 28 days after using the sample (P<0.05), with improvement rates of 7.27% and 12.73%, respectively. 5) Compared to before use, the level of wrinkles at the corners of the mouth in clinical evaluation was significantly reduced 14 and 28 days after using the sample (P<0.05), with improvement rates of 11.63% and 15.12%, respectively.

[0134] ●Subjective evaluation results: 1) After 28 days of using the sample, 96.97% of all subjects were satisfied with the sample. 2) After 28 days of sample use, 100% of subjects felt that the sample was mild and non-irritating.

[0135] ●Safety assessment results: Thirty-three subjects used the sample for 28 days, and no adverse skin reactions were observed.

[0136] Example 11 Transdermal compositions prepared in Examples 1 to 6 and 9: It is transparent and clear, (1) Particle size detection: Transmission electron micrographs showed particle sizes of 100 to 200 nm (see Figure 1). (2) Stability test: After centrifugation at 10,000 rpm for 5 minutes, no layer separation was observed, indicating excellent stability. (3) Water droplet performance in oil: For each sample, two equal volumes of emulsion (transdermal composition) were taken, and two drops of Sudan III dye and two drops of methylene blue dye solution were added to each at the same time, and the mixture was left to stand. The rate at which the two dyes (red for Sudan III and blue for methylene blue) diffused into the emulsion and the change in their appearance were observed.

[0137] Sudan III dyes are oil-soluble dyes and readily diffuse in oil phases, while methylene blue dyes are water-soluble dyes and readily diffuse in aqueous solutions.

[0138] The criterion for distinguishing the type of emulsion is that if the diffusion rate of blue is greater than that of red, it is an O / W emulsion; if not, it is a W / O emulsion.

[0139] Test results show that the transdermal composition prepared herein is a water-in-oil emulsion.

[0140] Comparative Examples 1 to 3 Comparative Examples 1 to 3 used other matrices, specifically as shown in Table 25. [Table 25]

[0141] As a result of the blending, it was found that the emulsifying effect of the comparative example was not as good as that of the example.

Claims

1. A transdermal composition comprising: Contains an active substance capable of exerting a predetermined effect, the transdermal composition is in the form of an emulsion; In the transdermal composition, calculated as a mass percentage of the total weight of the transdermal composition: the oil phase comprises 15% to 50%, preferably 30% to 45% or 35% to 40%, the emulsifier accounts for 35% to 65%, preferably 40% to 50%; A transdermal composition characterized in that the aqueous phase represents from 10% to 35%, preferably from 10% to 25%.

2. The transdermal composition according to claim 1, wherein the particle size distribution of the granules in the emulsion is 0.01 μm to 1 μm, or 100 to 200 nm.

3. By mass:

3. The transdermal composition according to claim 1, wherein the ratio of oil phase:water phase:emulsifier is 2-3:2.4-4:1 or 2-3:2.4-3.7:

1.

4. The components of the emulsion matrix, calculated as a percentage by mass of the total weight of the transdermal composition, are:

4. The transdermal composition according to any one of claims 1 to 3, characterized in that it comprises 1% to 15% oleic acid, 10% to 30% emulsion stabilizer, 5 to 15% of one or more of propylene glycol and butylene glycol, 0.5% to 4% menthol, 1% to 5% laurocapram, 35 to 65% emulsifier, and 10 to 20% water.

5. the emulsifier comprises sorbitan oleate and one or more selected from polysorbate 80, polysorbate 60, and polysorbate 20; Calculated as a mass percentage, The transdermal composition according to claim 4, wherein the sorbitan oleate accounts for 10-15% of the total weight of the transdermal composition, and the sum of any one or more of polysorbate 80, polysorbate 60, and polysorbate 20 accounts for 25-50% or 30-40% of the total weight of the transdermal composition.

6. The components of the emulsion matrix, calculated as a percentage by mass of the total weight of the transdermal composition, are:

6. A transdermal composition according to claim 4 or 5, comprising 1% to 15% oleic acid, 10% to 30% oleyl alcohol, 5% to 15% propylene glycol or butylene glycol, 0.5% to 4% or 1% to 2% menthol, 1% to 5% or 2% to 4% or 2% laurocapram, 10% to 15% sorbitan, 30% to 40% of any one or more of polysorbate 80, polysorbate 60 and polysorbate 20, and 10% to 20% water.

7. Contains preservatives, Preferably, the transdermal composition contains 0.1% to 1% by weight of a preservative, calculated as a percentage by mass of the total weight of the composition; The transdermal composition according to any one of claims 2 to 5, further characterized in that the preservative is selected from any one or more of phenoxyethanol, methylparaben, ethylparaben, propylparaben, benzyl alcohol and sorbic acid.

8. the active agent comprises a biomolecular agent; A transdermal composition according to any one of claims 1 to 6, characterized in that the biomolecular substance is preferably close to 300 kDa or less, or close to 150 kDa or less.

9. The biomolecular substance contained therein has a structure capable of being transported to a target cell and achieving the function of cleaving the exocytosis fusion of the target cell; Furthermore, the biomolecular substance is a clostridial neurotoxin, and preferably, the mass percentage of the clostridial neurotoxin in the total weight of the transdermal composition is 0.02% or less, more preferably 0.015% or less, and even more preferably 0.012% or less, and further, the clostridial neurotoxin is a botulinum toxin type A, B, C, D, E, F, or G, and / or the light chain of the clostridial neurotoxin is selected from the group consisting of SEQ ID NO:

9. The transdermal composition of claim 8, wherein the heavy chain of the clostridial neurotoxin comprises an amino acid sequence having at least 35%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 99% or 100% identity to any one of SEQ ID NOs: 1 to 7, and / or the heavy chain of the clostridial neurotoxin comprises an amino acid sequence having at least 35%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 99% or 100% identity to any one of SEQ ID NOs: 8 to 14.

10. The transdermal composition according to claim 8 or 9, characterized in that the components of the transdermal composition are as shown in Table 6.

11. The active substance is 10. The transdermal composition according to any one of claims 1 to 9, comprising any one or more of yeast extract, sodium ascorbyl phosphate, sea buckthorn seed oil, optional vitamins, retinyl palmitate, aloe vera extract, allantoin, licorice extract, Panax ginseng extract, Rheum palmatum extract, Centella asiatica extract, tea leaf extract and witch hazel extract, preferably the ingredients of the transdermal composition are as shown in Table 7.

12. Use of the transdermal composition according to any one of claims 1 to 11 in achieving therapeutic, cosmetic and skin care benefits.

13. The use according to claim 12, characterized in that when used, it is applied transdermally by one or more of the following methods: application, transdermal device, patch, and spray.

14. Use of the transdermal composition according to any one of claims 1 to 11 in the manufacture of a medicament or product for achieving therapeutic, cosmetic and skin care effects.

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