Macromolecular transdermal microemulsion system and application thereof

Abstract

The invention provides a macromolecular transdermal microemulsion system capable of enabling a macromolecular active substance to effectively perform a preset transdermal effect and application of the macromolecular transdermal microemulsion system. The macromolecular transdermal microemulsion system contains an effective amount of the macromolecular active substance capable of performing the preset effect. Wherein the macromolecular transdermal microemulsion system comprises the following components in percentage by mass based on the total weight of the transdermal microemulsion system: 7-36% of an oil phase, 6-12% of an emulsifying agent, 1-3% of an emulsifying agent and the balance of water; the proportion of the water phase is 57%-88%; the emulsion accounts for 8%-20%.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to a macromolecular transdermal microemulsion system and its applications. Background Technology

[0002] Transdermal drug delivery is a method of drug absorption through the skin. Drugs are absorbed through the skin into the bloodstream and reach effective blood drug concentrations to achieve disease treatment or prevention.

[0003] Transdermal drug delivery systems offer numerous advantages: they effectively reduce pain during injection, avoid irritation and side effects associated with oral medications, improve sustained-release effects, and enhance the safety of drug use and treatment. Transdermal drug delivery also avoids the first-pass effect in the liver, resulting in more stable blood drug concentrations and higher safety profiles.

[0004] However, human skin mainly functions as a barrier. This structure is conducive to "internal stability and external defense," protecting the human body from external stimuli and preventing the entry of foreign components. For foreign components to enter the skin, they must first penetrate the epidermis and enter the skin to exert their effects.

[0005] The skin consists of three parts: the epidermis, dermis, and subcutaneous tissue. The epidermis is the outermost layer of the skin, and it is divided into the stratum corneum, stratum lucidum, stratum granulosum, stratum spinosum, and stratum basale. The stratum basale is connected to the dermis via the basement membrane. The stratum corneum is the outermost layer of the epidermis, composed of 5–15 layers of keratinocytes and intercellular lipids; this structure is figuratively described as a "brick wall structure." The stratum corneum is 15–50 μm thick, has a very low water content (5%–20%), and is metabolically inactive, serving as the main barrier to the transdermal absorption of chemical substances. Intercellular lipids are mainly composed of 45%–50% ceramides, 25% cholesterol, 15% long-chain free fatty acids, and 5% other lipids. The dermis lies beneath the epidermis, providing support. It is mainly composed of connective tissue, including collagen fibers, elastic fibers, and matrix. The dermis also contains nerves, blood vessels, lymphatic vessels, muscles, hair follicles, sebaceous glands, and sweat glands, among other tissues.

[0006] Because of the dense stratum corneum structure of the skin, most drugs cannot be absorbed into the systemic circulation, and their bioavailability does not meet clinical requirements. This is especially true for large molecular weight bioactive drugs, which are even more difficult to exert their effects through the skin. Therefore, improving transdermal drug penetration is a key and challenging aspect of researching and developing transdermal drug delivery formulations.

[0007] Commonly used transdermal penetration enhancement techniques in physics include microneedles, iontophoresis, electroporation, ultrasound delivery, and needle-free drug delivery systems; commonly used chemical penetration enhancers include transdermal penetration enhancers and ion pairs.

[0008] Transdermal absorption enhancers are substances that promote the absorption of drugs through the skin, which is a preferred method. Currently, transdermal drug delivery systems have seen rapid development in drug development, with new products constantly being launched, such as estradiol and testosterone. In the cosmetic and medical aesthetics industry, some high-end cosmetics also develop emulsion-type transdermal formulations to achieve better results.

[0009] However, there is currently a lack of products that are effective for transdermal absorption of macromolecules. Summary of the Invention

[0010] This invention provides a macromolecular transdermal microemulsion system that enables macromolecular active substances to effectively perform predetermined effects through the skin and its applications.

[0011] Therefore, the present invention provides the following technical solution.

[0012] This invention provides a macromolecular transdermal microemulsion system, characterized in that it contains an effective amount of macromolecular active substances capable of performing a predetermined function; wherein, based on the mass percentage of the total weight of the transdermal microemulsion system, the composition of the macromolecular transdermal microemulsion system is as follows: oil phase: 7-36%; aqueous phase: 57%-88%; emulsion: 8%-20%.

[0013] The macromolecular transdermal microemulsion system provided by the present invention also has the following characteristics: wherein the oil phase accounts for 20%-30%, the aqueous phase accounts for 60%-70%, and the emulsion accounts for 8%-12%; and / or the composition of the transdermal microemulsion system by mass ratio is: oil phase: aqueous phase: emulsifier = 2-2.5:6-7:1.

[0014] The macromolecular transdermal microemulsion system provided by the present invention also has the following characteristics: wherein the macromolecular active substance is less than or equal to 300 kDa or less than or equal to 150 kDa, and / or, based on the mass percentage of the total weight of the transdermal microemulsion system, the effective amount of the macromolecular active substance in the transdermal microemulsion system is: the proportion of the macromolecular active substance is less than or equal to 10%, or less than or equal to 5%, or less than or equal to 2%, or less than or equal to 1%. Preferably, the macromolecular active substance is a biomolecule that can provide therapeutic, cosmetic, and skin care benefits; another preferred embodiment is that the macromolecular active substance is water-soluble.

[0015] The macromolecular transdermal microemulsion system provided by the present invention further has the following characteristics: wherein the macromolecular active substance contained therein is selected from at least one or more of clostridial neurotoxin, BSA, soluble collagen, and elastin, and / or is provided by yeast extract contained in the system; preferably, the clostridial neurotoxin is botulinum toxin of type A, B, C, D, E, F, or G, and / or the light chain of the clostridial neurotoxin contains an amino acid sequence that is at least 35%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% identical to any one of SEQ ID NO:1-7; and / or the heavy chain of the clostridial neurotoxin contains an amino acid sequence that is at least 35%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% identical to any one of SEQ ID NO:8-14.

[0016] The macromolecular transdermal microemulsion system provided by this invention also has the following characteristic: wherein, based on the mass percentage of the total weight of the transdermal microemulsion system, the system comprises:

[0017] The effective amount of the clostridium neurotoxin is less than or equal to 0.02%, preferably 0.00001%-0.015%, more preferably 0.0005%-0.015% or 0.0008-0.0015%; the effective amounts of soluble collagen, BSA and elastin are all less than or equal to 5%, or less than or equal to 2%, or less than or equal to 1%; the proportion of yeast extract is 1-10%.

[0018] The macromolecular transdermal microemulsion system provided by the present invention also has the following characteristics: the system comprises, by mass percentage of the total weight of the transdermal microemulsion system, 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 deacetylated chitosan or chitosan derivative; 55-80% or 60-70% water; and the effective amount of the macromolecular active substance.

[0019] The macromolecular transdermal microemulsion system provided by the present invention also has the following feature: wherein the emulsifier is selected from any one or more of polysorbate 80, polysorbate 60, polysorbate 20 and sorbitan oleate.

[0020] The macromolecular transdermal microemulsion system provided by the present invention also has the following characteristics: it contains a preservative, preferably, it contains 0.1-1% preservative by mass percentage of the total weight of the transdermal microemulsion system, and further, the preservative is selected from any one or more of phenoxyethanol, methylparaben, ethylparaben, propylparaben, benzyl alcohol and sorbic acid.

[0021] The macromolecular transdermal microemulsion system provided by the present invention also has the following feature: wherein the average particle size of the particles in the transdermal microemulsion system is 10-15 nm.

[0022] This invention also provides an application of a macromolecular transdermal microemulsion system in achieving therapeutic, cosmetic, and skincare effects.

[0023] The aforementioned application provided by the present invention is characterized in that: when used, it is applied transdermally through any one or more of the following methods: application, transdermal instrument, patch, and spray.

[0024] This invention also provides the application of a macromolecular transdermal microemulsion system in the preparation of drugs or products that achieve therapeutic, cosmetic, and skincare effects.

[0025] Invention Function and Effect

[0026] The macromolecular transdermal microemulsion system and its application provided by the present invention, due to the appropriate proportion of oil phase, water phase and emulsion in the composition of the macromolecular transdermal microemulsion system, forms a water-in-oil microemulsion with an average particle size of nanometers, which enables macromolecular active substances to be effectively absorbed through the skin and achieve the intended effect. Attached Figure Description

[0027] Figure 1 The standard curve for the BSA standard in Example 4 is fitted according to four parameters;

[0028] Figure 2 This is a phenotypic diagram of toe folding in the mouse experiment in Example 5. Detailed Implementation

[0029] The specific embodiments of the present invention will be described below with reference to the accompanying drawings. For the specific methods or materials used in the embodiments, those skilled in the art can make conventional substitutions based on the technical concept of the present invention and existing technologies, and are not limited to the specific descriptions of the embodiments of the present invention.

[0030] Unless otherwise specified, the methods used in the embodiments are conventional methods; the materials and reagents used are commercially available unless otherwise specified.

[0031] An emulsion is a non-uniformly dispersed liquid preparation consisting of two immiscible liquid phases, one of which is dispersed as small droplets within the other liquid phase. The phase forming the droplets is called the dispersed phase, internal phase, or discontinuous phase, while the other liquid phase is called the dispersion medium, external phase, or continuous phase.

[0032] An emulsion consists of an oil phase (represented by O), an emulsifier, and an aqueous phase (represented by W). Depending on the type, properties, and phase volume ratio (φ) of the emulsifier, it can form an oil-in-water (O / W) or water-in-oil (W / O) emulsion or a complex emulsion.

[0033] Aqueous phase: Water or aqueous solution. An aqueous solution refers to a raw material that is readily soluble in water.

[0034] Oil phase: refers to the raw material that is not easily soluble in water but is soluble in oily substances.

[0035] Based on the particle size of the emulsion particles, i.e. the size of the emulsion droplets, emulsions are classified into ordinary emulsions, submicron emulsions, and nanoemulsions:

[0036] 1. Regular milk: The droplets of regular milk are generally between 1μm and 100μm in size, and it is a milky white, opaque liquid.

[0037] 2. Subemulsions: Droplet sizes are generally between 0.1 μm and 1.0 μm. Subemulsions are often used as carriers for gastrointestinal drug delivery. Intravenous emulsions should be subemulsions, with particle sizes generally in the range of 0.25 μm to 0.4 μm.

[0038] 3. Nanoemulsions: The droplet size is <100nm, generally in the range of 10-100nm. Nanoemulsions are also called microemulsions.

[0039] The macromolecular transdermal microemulsion system provided by this invention contains an effective amount of macromolecular active substances that can perform a predetermined function; wherein, based on the mass percentage of the total weight of the transdermal microemulsion system, the composition of the macromolecular transdermal microemulsion system is as follows: oil phase: 16-36% or 20-30%; aqueous phase: 55%-88% or 60-70%; emulsion: 8%-20% or 10%-25%.

[0040] Macromolecular active substances refer to substances with large molecular weights, mainly biomolecules.

[0041] Da, short for Dalton, is a commonly used unit of molecular weight. It is the algebraic sum of the atomic weights of all atoms in a molecule. Numerically, Dalton is equal to the relative molecular mass.

[0042] In biochemistry, molecular biology, and proteomics, kDa (kilodaltons) is frequently used to represent biological macromolecules such as proteins. 1 kDa represents a molecule with a relative molecular mass of 1000. Macromolecules refer to biological substances with a relative molecular mass of 5000 or more, even exceeding one million, such as polypeptides, proteins, nucleic acids, polysaccharides, and antibodies.

[0043] The composition of the macromolecular transdermal microemulsion system provided by this invention is shown in Table 1:

[0044]

[0045] Table 1 refers to the composition of the macromolecular transdermal microemulsion system of the present invention, which is divided into three types: oil phase, aqueous phase and emulsifier. The "effective amount of macromolecular active substance that can play a predetermined role" may be oil-based (soluble in oil) or aqueous (soluble in water).

[0046] In one example, the macromolecular active substance of the present invention refers to a biomolecule with a value of less than or equal to approximately 300 kDa or less than or equal to approximately 150 kDa. Preferably, the biomolecule is water-soluble, that is, it exists in the aqueous phase after being added.

[0047] In one example, the intended effect is to achieve one or more of the following effects: therapeutic, cosmetic, and skincare effects.

[0048] In one example, the macromolecular transdermal microemulsion system of the present invention is defined to satisfy any one or two of the conditions shown in Table 2:

[0049]

[0050] The macromolecular active substance only needs to be able to function through this system. Based on the mass percentage of the total weight of the transdermal microemulsion system, the effective amount of the macromolecular active substance in the transdermal microemulsion system is: less than or equal to 10%, or less than or equal to 5%, or less than or equal to 2%, or less than or equal to 1%. Within this range, adjustments can be made according to the specific macromolecular active substance. After final adjustment, the overall oil phase, aqueous phase, and emulsifier of the microemulsion system should meet the aforementioned ratio or proportion requirements.

[0051] In one example, the macromolecular active substance contained is selected from at least one or more of clostridium neurotoxin, soluble collagen, and elastin, and / or provided by yeast extract contained in the system: that is, the macromolecular active substance contained can be one of the following types as shown in Table 3:

[0052]

[0053] In one example, for any of the cases in Table 3, the content, as a percentage of the total weight of the transdermal microemulsion system, is as follows:

[0054] The effective amount of the clostridium neurotoxin is less than or equal to 0.02%, preferably 0.00001%-0.015%, more preferably 0.0005%-0.015% or 0.0008-0.0015%, and the content of the clostridium neurotoxin is 10 ng / ml-10 μg / ml;

[0055] The effective amounts of soluble collagen and elastin are both less than or equal to 5%, or less than or equal to 2%, or less than or equal to 1%.

[0056] The effective amount is provided by yeast extract, which accounts for 1-10% of the content in the emulsion system. For example, if the yeast extract accounts for 10% and contains 5% macromolecules, then the effective amount is 5%.

[0057] Clostridium toxins have therapeutic and cosmetic applications. For example, botulinum toxin specifically binds to presynaptic receptors at peripheral cholinergic nerve endings, cleaving the synaptic-associated protein SNAP-25, interfering with the exocytosis of presynaptic vesicles, and inhibiting the release of acetylcholine (ACh) from nerve endings. This induces chemical denervation of muscles, leading to muscle relaxation and relief of muscle spasms, thus exerting a therapeutic effect. The muscle chemical denervation effect of botulinum toxin is the theoretical basis for regulating hypertonia, movement disorders, and dynamic wrinkles. Furthermore, repeated injections into target muscles can cause disuse atrophy, reducing muscle volume; therefore, botulinum toxin is also used for body sculpting. Currently, botulinum toxin is used in the treatment of diseases including hemifacial spasm, idiopathic blepharospasm, spasmodic torticollis, spastic cerebral palsy, post-stroke limb spasticity, tremor, and other types of dystonia. Botulinum toxin is used in minimally invasive cosmetic procedures for various dynamic wrinkles, including frown lines, forehead lines, and crow's feet; facial contour enhancement, such as adjusting eyebrow height, relaxing the mentalis muscle, lifting the jawline, and injecting nerve strips; and body contouring, such as reducing the masseter muscle, gastrocnemius muscle, and trapezius muscle.

[0058] The term "clostridium toxins" in this article refers to natural clostridium toxins, as well as toxins with similar structures and functions to natural clostridium toxins.

[0059] Although the amino acid sequences and immunogenicity of various types of toxins differ, they all exhibit similar molecular structures. Clostridium neurotoxins are produced by toxin-producing Clostridium as a non-toxic single-chain polypeptide, approximately 150 kDa. They become active only after being cleaved into a double-chain form by bacterial proteases or in vitro proteases. This double-chain form consists of a light chain (L chain, amino terminus of the toxin, 50 kDa) and a heavy chain (H chain, carboxyl terminus of the toxin, 100 kDa) linked by a disulfide bond. The heavy chain is further composed of two domains: Hn (amino terminus, 50 kDa) and Hc (carboxyl terminus, 50 kDa).

[0060] Furthermore, the clostridial neurotoxin is botulinum toxin of type A, B, C, D, E, F, or G, and / or the light chain of the clostridial neurotoxin contains an amino acid sequence that is at least 35%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% identical to any one of SEQ ID NO: 1-7; and / or the heavy chain of the clostridial neurotoxin contains an amino acid sequence that is at least 35%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% identical to any one of SEQ ID NO: 8-14. The sequences are shown in Table 4.

[0061]

[0062]

[0063]

[0064]

[0065] Botulinum toxin-related products have broad application prospects in the treatment of clinical spastic diseases, glandular hypersecretion, neuropathic pain, and cosmetic wrinkle removal. However, due to their large molecular size, these products are currently administered via injection, making it difficult for them to directly penetrate the skin and reach the target site to exert their effects.

[0066] Currently, only botulinum toxin types A and B are available as pharmaceuticals, but the vast majority of botulinum toxin used in the medical field is type A. Clinically used type A botulinum toxins include Botox produced by Allergan (USA), Dysport produced by Speywood (UK), and Hengli (BTX-A) produced by the Lanzhou Institute of Biological Products in my country. Botulinum toxin-related products have broad application prospects in the treatment of spastic diseases, hypersecretion of glands, neuropathic pain, and cosmetic wrinkle removal. However, these products are currently administered via injection. Our large-molecule transdermal microemulsion system, through pharmacodynamic experiments in mice, has demonstrated that it enables large-molecule biomolecules such as botulinum toxin to achieve their intended effects through transdermal delivery.

[0067] Yeast extract (also known as yeast extract, abbreviated as YE) is a pure natural product made from protein-rich edible yeast using modern biotechnology such as autolysis, enzymatic hydrolysis, separation, and concentration. It is produced by degrading the proteins and nucleic acids within the yeast cells. The extract is a brownish-yellow soluble paste or light yellow powder. In addition to amino acids and nucleotides, yeast extract also contains monosaccharides, polysaccharides, minerals, and vitamins.

[0068] The main applications of yeast extract are skin rejuvenation, anti-aging, and moisturizing. Yeast extract has a good skin rejuvenation effect, which is achieved through three main mechanisms: (1) reducing the transfer of melanin to surface cells; (2) promoting the metabolism of melanin-containing keratinocytes and accelerating the shedding of keratinocytes; and (3) promoting the synthesis of epidermal proteins and regulating skin texture. Yeast extract has a good effect in scavenging free radicals and anti-aging, which can regulate skin texture, fine lines and wrinkles, increase skin elasticity, make the skin smooth and delicate, and reduce and prevent dull skin tone problems caused by early aging. Yeast extract also has a strong moisturizing effect: it is rich in natural moisturizing factors, which enable the stratum corneum to retain moisture more effectively, give the skin deep hydration, and prevent dry skin; it strengthens the skin barrier function by increasing the content of ceramides in the stratum corneum and reducing transepidermal water loss; it can also repair the damaged lipid barrier of the stratum corneum, improve skin resistance, and regulate skin condition.

[0069] Soluble collagen is a natural protein that plays an important role in the skin. Collagen maintains the skin's elasticity and firmness, but as we age, the amount of collagen in the skin gradually decreases, leading to sagging skin and wrinkles. Soluble collagen, as a form that can penetrate deeper into the skin, can replenish collagen and thus improve the skin's condition.

[0070] Soluble collagen can stimulate and promote collagen synthesis, help repair damaged collagen, and promote the generation of new collagen, thereby improving skin elasticity and firmness, and reducing wrinkles and fine lines.

[0071] Elastin is an essential component of skin, and it's considered the foundation of youthful skin. Elastin peptides promote skin damage repair, angiogenesis, and fibroblast proliferation. They also enhance skin elasticity, resulting in smooth and supple skin.

[0072] In one example, the composition of the macromolecular transdermal microemulsion system of the present invention is shown in Table 5:

[0073]

[0074]

[0075] Preferably, the example of Table 5 is composed of Table 6:

[0076]

[0077] Chitosan, a natural alkaline polysaccharide, possesses film-forming, antibacterial, anti-inflammatory, hemostatic, analgesic, antipruritic, scar-reducing, wound-healing-promoting, and antioxidant properties. Chitosan exhibits excellent inhibitory effects on bacteria, yeast, fungi, and other microorganisms. It significantly inhibits common human epidermal bacteria such as Staphylococcus epidermidis, Escherichia coli, and Candida tropicalis, as well as infections common in burn patients such as Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus pyogenes. Chitosan has powerful moisturizing properties, even surpassing sodium hyaluronate, earning it the title of "super moisturizer." Chitosan can adsorb heavy metals and residues from cosmetics. Furthermore, chitosan possesses repairing properties, helping to repair the skin barrier and is beneficial for sensitive and acne-prone skin.

[0078] In one example, the macromolecular transdermal microemulsion system also contains a preservative, preferably 0.1-1% by mass percentage of the total weight of the transdermal microemulsion system. Further, the preservative is selected from any one or more of phenoxyethanol, methylparaben, ethylparaben, propylparaben, benzyl alcohol, and sorbic acid.

[0079] The average particle size of the macromolecular transdermal microemulsion system of the present invention is in the nanometer range, preferably 10-15 nm.

[0080] Table 5 is a specific example of the composition of the macromolecular transdermal microemulsion system provided by the present invention.

[0081]

[0082]

[0083] The macromolecular transdermal microemulsion system of the present invention is used to achieve therapeutic, cosmetic and skin care effects. When used, it can be applied through one or more of the following methods: application, transdermal instrument, patch, and spray.

[0084] The macromolecular transdermal microemulsion system of the present invention can also be used in the preparation of drugs or products that achieve therapeutic, cosmetic and skin care effects.

[0085] In addition, in this invention, the water is preferably any one or a combination of distilled water, drinking water, sterile water, ultrapure water, and deionized water.

[0086] In the following embodiments, some components are described below:

[0087] The main applications of superoxide dismutase (SOD) in cosmetics include: (1) as a cosmetic additive, it can prevent skin aging and play a skin care role; (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 masks, lotions and creams.

[0088] Vitamins are essential organic substances for the human body. Vitamin A can promote the proliferation of epidermal cells and increase the formation of dermal collagen and elastin, thus having anti-aging effects. Vitamin E is a natural antioxidant that can promote metabolism and improve skin elasticity. Vitamin C has strong antioxidant properties, promoting collagen synthesis and inhibiting collagen breakdown. Vitamins C and E have a synergistic effect in scavenging free radicals.

[0089] In addition to essential vitamins, many natural plant-based active ingredients have been proven to have anti-wrinkle and firming functions, such as Centella asiatica extract, seaweed extract, and calendula extract. These ingredients are now widely used in anti-wrinkle and firming cosmetics.

[0090] Example 1

[0091] The formulation of the macromolecular transdermal microemulsion system provided in this embodiment is shown in Table 6:

[0092]

[0093] Preparation method:

[0094] (1) Using an electronic balance, weigh the purified water, carboxymethyl deacetylated chitosan and botulinum toxin A into a 3L beaker 1 according to the formula amount, and stir to dissolve at room temperature (speed: 400-500 rpm).

[0095] (2) Using an electronic balance, weigh out the polysorbate-80, laurocapram, phenoxyethanol, propylene glycol and glycerin in sequence according to the formula into a 2L beaker 2, and stir evenly at room temperature (speed: 250-300 rpm).

[0096] (3) Add the solution in beaker 2 to beaker 1 and stir evenly at room temperature (speed: 400-500 rpm).

[0097] (4) Add purified water to the solution prepared in step 3) to make up to 1000g, and stir at room temperature.

[0098] Example 2

[0099] The formulation of the macromolecular transdermal microemulsion system provided in this embodiment is shown in Table 7.

[0100]

[0101] Preparation method:

[0102] (1) Using an electronic balance, weigh the purified water, carboxymethyl deacetylated chitosan, yeast extract, superoxide dismutase (SOD) and sodium ascorbate phosphate into a 3L beaker 1 according to the formula amount, and stir to dissolve at room temperature (speed: 400-500 rpm).

[0103] (2) Using an electronic balance, weigh out the following ingredients in sequence according to the formula: polysorbate-80, tocopherol (vitamin E), retinyl palmitate, phenoxyethanol, (daily) fragrance, butylene glycol and glycerin into a 2L beaker 2, and stir evenly at room temperature (speed: 250-300 rpm).

[0104] (3) Add the solution in beaker 2 to beaker 1 and stir evenly at room temperature (speed: 400-500 rpm).

[0105] (4) Add purified water to the solution prepared in step (3) to make up the difference, and bring to room temperature.

[0106] Stir and mix well.

[0107] Example 3

[0108] The formulation of the macromolecular transdermal microemulsion system provided in this embodiment is shown in Table 8.

[0109]

[0110]

[0111] Preparation method:

[0112] (1) Using an electronic balance, weigh the purified water, hydroxypropyl tetrahydropyrantriol, carnosine, sodium polyglutamate, biotin, seaweed extract, calendula extract, centella asiatica extract, aloe vera extract, carboxymethyl deacetylated chitosan, sodium ascorbate phosphate, yeast extract, superoxide dismutase (SOD), soluble collagen, and elastin into a 3L beaker 1 and stir to dissolve at room temperature (speed: 400-500 rpm).

[0113] (2) Using an electronic balance, weigh out the following ingredients in sequence according to the formula: polysorbate-80, tocopherol (vitamin E), retinyl palmitate, phenoxyethanol, (daily) fragrance, butylene glycol and glycerin into a 2L beaker 2, and stir evenly at room temperature (speed: 250-300 rpm).

[0114] (3) Add the solution in beaker 2 to beaker 1 and stir evenly at room temperature (speed: 400-500 rpm).

[0115] (4) Add purified water to the solution prepared in step (3) to make up to 1000g, and stir at room temperature.

[0116] The formulations obtained in Examples 1-3 have good emulsification effect, high transparency, and no stratification. The average particle size was measured to be 10-15 nm by a nanoparticle size potentiometer. In particular, the formulation obtained in Example 3 had an average particle size of 12.99 nm.

[0117] Comparative Example 1

[0118] The formulations for this comparative example are shown in Table 9.

[0119]

[0120] Preparation method:

[0121] (1) Add the ester to a 2L beaker and stir for 5 minutes; weigh the oleyl alcohol and oleic acid into the beaker according to the formula using an electronic balance and stir for 5 minutes; weigh the laurocapram and phenoxyethanol into the beaker according to the formula using an electronic balance and stir evenly at room temperature (speed: 200-250 rpm, 5 minutes).

[0122] (2) Weigh propylene glycol and menthol according to the formula using an electronic balance and add them to the blue-capped bottle in sequence. Heat to 60°C (about 10 min) to fully dissolve them, then add them to the solution obtained in step 1). Stir evenly at room temperature (speed: 200-250 rpm, 5 min).

[0123] (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).

[0124] (4) Add purified water to the solution prepared in step (3) to make up to 1000g, and stir at room temperature.

[0125] Comparative Examples 2-10

[0126] The formulations for Comparative Examples 2-10 are shown in Table 10, by mass percentage:

[0127]

[0128]

[0129] According to the formulations in Comparative Examples 2-10, the oil phase is thoroughly mixed and dissolved, and the water phase is thoroughly mixed and dissolved, and then the phases are mixed to obtain the final product. The specific preparation methods are all conventional techniques that can be selected and can be referred to in the previous examples, and will not be described one by one.

[0130] The comparative examples 2-10 prepared showed inferior appearances compared to examples 1-3, as detailed in Table 11:

[0131]

[0132] Example 4

[0133] This experiment was conducted to detect transdermal efficiency.

[0134] 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 had a concentration of 1.0 mg / ml, and the BSA protein emulsion prepared in Comparative Example 1 had a concentration of 0.62 mg / ml. Transdermal experiments were conducted using the Franz diffusion cell method.

[0135] Test equipment and materials

[0136] 1. Reagents

[0137] Phosphate buffer, protein standards, test samples, protein detection kits (ultra-high sensitivity);

[0138] 2. Consumables

[0139] Transdermal evaluation was conducted using excised porcine skin, centrifuge tubes, EP tubes, pipette tips, and pre-coated microplates; 3. Equipment

[0140] 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.), pipette (Eppendorf AG).

[0141] I. Preparations before the experiment:

[0142] (1) The pig skin required for the experiment was taken out of the -20℃ freezer and thawed at 4℃, then washed with physiological saline and set aside.

[0143] (2) Clean the supply chamber, diffusion chamber (receiving chamber) and stir bar before the experiment.

[0144] (3) Add pure water to the water tank, making sure it is level with the diffuser frame, and set the temperature to 32℃ and the rotation speed.

[0145] (4) After the temperature of the TP-6 ​​transdermal diffusion instrument reaches the set temperature, place the pigskin between the supply chamber and the diffusion chamber. Stir at a constant speed to keep the solution in an isothermal state and to ensure that the concentration of the exudate is uniform.

[0146] II. Experimental Methods:

[0147] (1) Set up 3 experimental groups and one blank group.

[0148] (2) Add BSA protein test solution: Add 150 μl (1 mg / ml) of BSA protein emulsion test solution prepared in Example 1, 150 μl (0.62 mg / ml) of BSA protein emulsion test solution prepared in Comparative Example 1, and 150 μl (1 mg / ml) of BSA protein test solution dissolved in PBS to the first 3 supply chambers respectively. Add 150 μl of PBS to the blank group.

[0149] (3) Fill the receiving chamber with PBS. Take four 1.5ml EP tubes and take 400ul of liquid from the receiving chamber every 1h for subsequent OD detection. Mark the sampling time and experimental group number. Sampling time is 6h in total. After sampling, each receiving chamber must be replenished with 400μl of PBS to maintain the volume of the receiving chamber.

[0150] (4) Place all collected samples in a 4°C refrigerator for later use.

[0151] III. ELISA Experimental Detection:

[0152] 1. Reagent preparation:

[0153] a. Equilibrate all reagents to room temperature (18-25℃) before use. Set the detection wavelength to 450nm according to the microplate reader's instruction manual and preheat the plate for 15 minutes before reading.

[0154] b. Washing buffer: Dilute 20 ml of concentrated washing buffer with 580 ml of deionized water to prepare 600 ml of washing buffer.

[0155] c. Standard Working Solution: First, centrifuge the standard at 1000g for 1 min, add 2ml of standard sample diluent, and mix thoroughly. Pipette 300μl of the 20000ng / ml standard diluent into the first tube and mix again to obtain the 6666.7ng / ml working solution. Then transfer 300μl of the solution from the first tube to the second tube. Before the next transfer, thoroughly mix each tube. Set six dilution points for the standard: 20000ng / ml, 6666.7ng / ml, 2222.2ng / ml, 740.7ng / ml, 246.9ng / ml, and 0ng / ml.

[0156] d. Working solution of reagent A: Calculate the required volume (100 μl / well) before the experiment. During preparation, prepare 100-200 μl more than the calculated volume. Before use, slightly centrifuge the stock solution tube and dilute 100× concentrated reagent A to 1× working solution A with reagent diluent A (e.g., 10 μl reagent A + 990 μl reagent A diluent).

[0157] e. Working solution of reagent B: Calculate the required volume (100 μl / well) before the experiment. During preparation, prepare 100-200 μl more than the calculated volume. Before use, slightly centrifuge the stock solution tube and dilute 100× concentrated reagent B to 1× working solution B with reagent diluent B (e.g., 10 μl reagent B + 990 μl reagent B diluent).

[0158] 2. Experimental Methods

[0159] (1) Set up standard wells, sample wells, and blank wells respectively. Add 6 standards of different concentrations (including the zero well, 50 μL / well) in sequence, and add 50 μL of the test sample (the sample collected from the sampling port of the receiving chamber) to the other wells. Then immediately add 50 μL of working solution of test reagent A per well, gently shake to mix, cover with the sealing film provided by the kit, and incubate at 37°C for 60 minutes.

[0160] (2) Discard the liquid in all wells, add 350 μL 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.

[0161] (3) Add 100 μL of detection reagent B working solution to each well, cover with sealing film, and incubate at 37°C.

[0162] 30 minutes.

[0163] (4) Discard the liquid in each hole and repeat the washing process in step 2 5 times.

[0164] (5) Add 90 μL of TMB reagent to each well, cover with a new sealing film, and incubate at 37°C in the dark.

[0165] 10-20 minutes.

[0166] (6) Add 50 μL of stop solution to each well, in the same order as the addition of colorimetric solution, and gently shake to mix.

[0167] (7) Ensure there are no air bubbles or water vapor at the bottom of the wells of the ELISA plate, and immediately measure the absorbance of each well at 450 nm.

[0168] Measure the OD value and record the microplate reader reading.

[0169] IV. Test Data and Conclusions

[0170] (1) The BSA standard is fitted with a standard curve according to four parameters, as shown in the figure. Figure 1 As shown.

[0171] Plotting the standard curve:

[0172] Processing the OD value data of the detected standard proteins:

[0173] The data was fitted using four parameters to obtain the standard curve. The four-parameter fitting formula is as follows:

[0174]

[0175] For specific standard curves, see [link to standard curve]. Figure 1 .

[0176] (2) Sample test results

[0177] Table 12 shows the concentration of BSA transmitted through the skin at different time periods for different test samples, converted according to the standard curve.

[0178]

[0179] Based on the results, transdermal calculations were performed:

[0180] (1) After 24 hours, the concentration of BSA emulsion protein test solution prepared in Example 1 was 239.6 ng / ml in the receiving chamber and its volume was 15 ml, while the concentration of BSA in the supply chamber was 1 mg / ml and the volume added in the supply chamber was 150 μl. Therefore, the transdermal efficiency of the BSA emulsion prepared in Example 1 was calculated to be 2.40%.

[0181] (2) Comparative Example 2 prepared BSA emulsion protein test solution. After 24 hours, the concentration in the receiving chamber was 147.1 ng / ml and the volume was 15 ml. The concentration of BSA in the supply chamber was 0.62 mg / ml and the volume added in the supply chamber was 150 μl. Therefore, the transdermal efficiency of the BSA emulsion prepared in Example 5 was calculated to be 2.37%.

[0182] (3) The BSA protein test solution dissolved in PBS was 125.4 ng / ml in the receiving chamber after 24 hours, and its volume was 15 ml. The BSA concentration in the supply chamber was 1 mg / ml, and the volume added to the supply chamber was 150 μl. Therefore, the transdermal efficiency of PBS was calculated to be 1.25%.

[0183] It is evident that, compared to the control, both Example 1 and Comparative Example 2 exhibit excellent transdermal effects, with Example 1 demonstrating even better transdermal effects than Comparative Example 2.

[0184] Example 5

[0185] The efficacy verification experiment of botulinum toxin protein A microemulsions with a concentration of 10 μg / ml prepared in Example 1 and Comparative Example 1 in mice.

[0186] To compare the differences in transdermal efficiency of macromolecular active substances between Example 1 and Comparative Example 1, we used 17-19g SPF-grade CD-1 (ICR) mice. 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 dipped in 10% alcohol, dried, and then the medication was applied. 200μl of botulinum toxin A microemulsion was applied to the skin of the mouse's right hind leg once daily for 7 consecutive days. Botulinum toxin A was not applied to the left hind leg of the mice.

[0187] The experimental results showed that 2-3 days after applying botulinum toxin A, mice developed a symptom of fused toes on their right hind legs (2-5 toes fused together). The experimental results of Example 1 and Comparative Example 1 are shown in Table 13. The emulsion prepared in Example 1 showed fused toes as early as D2: 33% of the experimental mice had two fused toes and 33% had five fused toes; D3: 33% of the experimental mice had two fused toes and 33% had five fused toes; D4: 66% of the experimental mice had three fused toes and 33% had five fused toes; D5: 100% of the experimental mice had five fused toes.

[0188] The emulsion prepared in Comparative Example 1 showed no syndactyly in D2; syndactyly in D3: 33% of the experimental mice had two syndactyly; syndactyly in D4: 66% of the experimental mice had two syndactyly, and 33% of the experimental mice had three syndactyly; syndactyly in D5: 33% of the experimental mice had three syndactyly, and 66% of the experimental mice had four syndactyly.

[0189]

[0190] The results in Table 13 show that the botulinum toxin A microemulsion prepared in Example 1 had significantly better efficacy in mice than that in Comparative Example 1. The microemulsion of Example 1 not only acted earlier but also had better efficacy than that of Comparative Example 1. The toe-clamping phenotypes of Example 1 and Comparative Example 1 are shown below. Figure 2 As shown.

[0191] According to literature reports, mice injected with botulinum toxin A into their gastrocnemius muscle exhibited a symptom of their toes closing together.

[0192] As can be seen, by using the emulsion delivery system we developed to make botulinum toxin A into a microemulsion and applying it to the skin of mice, we achieved the syndactyly phenotype after botulinum toxin A injection. This demonstrates that our developed emulsion system is more effective in mice.

[0193] Example 7

[0194] The efficacy of the anti-wrinkle and firming cosmetic microemulsion prepared in Example 3 in the human body was tested.

[0195] 1. Under normal circumstances, adult subjects will use the product continuously for 28 days according to the instructions to evaluate whether the product has moisturizing, repairing, firming, and anti-wrinkle effects, and whether the product is suitable for sensitive skin and is gentle and non-irritating.

[0196] 2. Participants: A total of 31 valid participants completed the assessment. They were healthy Chinese women with sensitive skin (screened using SGS's internal sensitive skin questionnaire), aged 31 to 60 years, with a mean age of 52.65 ± 6.08 years, meeting the inclusion and exclusion criteria. The assessment area was the face.

[0197] Inclusion and exclusion criteria for participants:

[0198] Participants in this evaluation were selected from SGS's CPCH efficacy laboratory participant information database, choosing healthy participants who met the following inclusion criteria and those who did not meet the following exclusion criteria.

[0199] (1) Selection criteria

[0200] Healthy women, aged 28-60;

[0201] Race: Asian (China);

[0202] The subjects had sensitive skin on their faces (screened through SGS's internal sensitive skin questionnaire);

[0203] Subjects rated their own discomfort symptoms, such as (non-persistent) itching or stinging sensation on a scale of 4-7 (0-9 scale, self-assessment questionnaire).

[0204] The subjects perceived their facial skin as dull, loose, and lacking elasticity;

[0205] Forehead wrinkles visual score 3-6 (based on SGS internal map);

[0206] Visual score of 3-6 for lateral canthoplasty wrinkles on one side (based on SGS internal atlas);

[0207] One-sided nasolabial fold wrinkle (laugh lines) visual score 3-6 (according to SGS internal atlas);

[0208] Transepidermal water loss (TEWL) in one cheek area >15 g / h / m2;

[0209] The measured moisture content of the stratum corneum in one cheek area was <60 a.u.;

[0210] The facial skin showed no obvious damage, scars, or hair loss.

[0211] They can cooperate well with the evaluation project according to the requirements of the plan and maintain a regular lifestyle during the research period.

[0212] (2) Exclusion criteria

[0213] Anyone meeting any of the following conditions must be excluded from this study:

[0214] Those who do not agree to sign the informed consent form;

[0215] Those who are unwilling to comply with the requirements of the plan;

[0216] Simultaneously participate in any other clinical research;

[0217] Assess the use of cosmetics and / or medications on the day of the event;

[0218] She stated that she was pregnant and breastfeeding.

[0219] He is currently receiving medication during the research period;

[0220] Subjects suffering from infectious skin diseases or atopic dermatitis;

[0221] Those with skin abnormalities such as moles or telangiectasia in the assessment area;

[0222] Subjects who have undergone skin peeling or skin treatment within 3 months prior to participating in the assessment;

[0223] Subjects who received immunosuppressant therapy within 3 months prior to participating in the evaluation;

[0224] Subjects who have received systemic steroid treatment or phototherapy within one month prior to participating in the assessment;

[0225] Two weeks prior to the assessment, the user used topical medications or / and cosmetics with special effects (claiming to moisturize, repair, firm, or anti-wrinkle) on the affected area.

[0226] The area being assessed has lesions, obvious traces, or other abnormalities, making measurement difficult;

[0227] Subjects who have severe reactions or allergies to cosmetics, drugs or general light exposure;

[0228] In addition to the above, if the project leader deems it unsuitable to conduct an evaluation.

[0229] 3. How to use: After cleansing, thoroughly remove oil and keep skin dry. Take an appropriate amount of serum (microemulsion), drop two drops from the dropper, and apply to areas with wrinkles such as the corners of the eyes, forehead, nasolabial folds, and chin. Massage until the serum (microemulsion) is absorbed. It is recommended to use daily, 7 days a week. A 4-week usage cycle is recommended (when not in use, please store the product in the refrigerator at a constant temperature of 2-8℃).

[0230] 4. Evaluation period: before product use (D0), 14 days after product use (D14), and 28 days after product use (D28).

[0231] 5. Evaluation parameters:

[0232] (1) Image acquisition

[0233] Primos CR is used for facial image acquisition and photo analysis of skin wrinkles. A smaller analysis value indicates an improvement in skin wrinkles.

[0234] (2) Skin elasticity

[0235] Skin elasticity tester The MPA580 is used to test skin elasticity. Increased R2, R5, and R7 values ​​indicate improved skin elasticity.

[0236] (3) Skin firming

[0237] Skin elasticity tester The MPA580 is used to test skin firmness. A smaller F4 measurement indicates improved skin firmness.

[0238] (4) Moisture content of the stratum corneum of the skin

[0239] Skin moisture content meter The CM825 is used to detect the moisture content of the stratum corneum of the skin. A higher measurement indicates an increase in the moisture content of the stratum corneum.

[0240] (5) Transepidermal water loss rate

[0241] Skin moisture loss meter TM Hex is used to measure transepidermal water loss from the skin. A smaller measured value indicates an improved skin barrier.

[0242] (6) TC value

[0243] The TC value is a secondary parameter in non-invasive testing, representing the amount of transepidermal water loss per unit area per unit time. A smaller TC value indicates improved skin barrier function.

[0244] (7) Skin moisture distribution map

[0245] The MoistureMap MM 200 skin moisture distribution meter is a unique instrument based on capacitance imaging, used to observe hydration distribution and texture characteristics. MGL represents the average gray value of skin moisture distribution; the lower the value, the higher the moisture content.

[0246] (8) Skin luster

[0247] Skin gloss meter The GL200 is used to measure skin gloss. A higher measurement indicates increased skin gloss.

[0248] (9) Subject self-assessment

[0249] Participants conducted self-assessments based on their own usage.

[0250] 6. Evaluation scheme design:

[0251] (1) Before using the product (D0):

[0252] Upon arrival at SGS, participants cleansed their faces with facial cleansing products and dried their skin with lint-free dry facial tissues. They sat quietly for 30 minutes in a laboratory at a temperature of 21±1℃ and 50±10% RH. A dermatologist conducted a visual assessment, and those who met the inclusion criteria proceeded to the next stage of evaluation.

[0253] Laboratory technicians measured the facial skin stratum corneum moisture content (Corneometer) and transepidermal water loss rate (Tewameter). Those meeting the inclusion criteria proceeded to the next stage of evaluation.

[0254] Laboratory technicians collected Primos CR images, Moisture Map MM200, Cutometer, and Glossymeter measurements from qualified subjects.

[0255] (2) Subjects complete questionnaires

[0256] Laboratory technicians explained the product usage instructions to the subjects and distributed the products. Subjects then tried the samples on-site under the supervision of the laboratory technicians. If any adverse reactions occurred, subjects were required to promptly fill out a usage log.

[0257] (3) 14 days after using the product (D14)

[0258] After arriving at SGS, the subjects cleaned their faces with facial cleansing products and dried their skin with lint-free dry facial tissues. They sat quietly for 30 minutes in the laboratory at a temperature of 21±1℃ and 50±5% RH. Laboratory technicians collected Primos CR images of the subjects' faces and measured the facial skin with Cutometer, Moisture Map MM200, Tewameter, Corneometer, and Glossymeter.

[0259] Laboratory technicians weighed the products and checked the product usage logs.

[0260] The subject left SGS.

[0261] (4) 28 days after using the product (D28)

[0262] After arriving at SGS, the subjects cleaned their faces with facial cleansing products and dried their skin with lint-free dry facial tissues. They sat quietly for 30 minutes in the laboratory at a temperature of 21±1℃ and 50±5% RH. Laboratory technicians collected Primos CR images of the subjects' faces and measured the facial skin with Cutometer, Moisture Map MM200, Tewameter, Corneometer, and Glossymeter.

[0263] Participants completed and used questionnaires;

[0264] Laboratory technicians weigh and collect the products, and check and collect the product usage logs;

[0265] The subject left SGS.

[0266] (5) Data Statistics: Data were analyzed using SPSS 28.0. A normality test was performed on the evaluation data. If the evaluation data were normally distributed, a t-test was used for statistical analysis; if the evaluation data were not normally distributed, a rank-sum test was used for statistical analysis. Rank-sum tests were used for ordinal data. The significance level for statistical methods was P < 0.05.

[0267] Evaluation results, based on 28 days of continuous product use by 31 healthy Chinese women with sensitive skin, indicate that under these evaluation conditions, the product demonstrated moisturizing, repairing, firming, and anti-wrinkle effects within 14 days, and was suitable for sensitive skin, being gentle and non-irritating. Specific results are as follows:

[0268] (1) Instrument evaluation results (Table 14)

[0269]

[0270]

[0271]

[0272] (2) Subject self-assessment (satisfaction)

[0273] After using the product for 14 days, 94% of the participants felt it deeply moisturized, 94% felt their skin was hydrated and plump, 94% felt it had a good moisturizing effect, 90% felt their skin was firmer, 87% felt their skin became more elastic, 87% felt their wrinkles improved, 87% felt their fine lines improved, 87% felt their overall skin condition improved, 90% felt the product was suitable for sensitive skin, and 100% felt the product was gentle and non-irritating.

[0274] After 28 days of product use, 100% of the participants felt the product deeply hydrated, 100% felt their skin was plump and hydrated, 100% felt it had a good moisturizing effect, 100% felt their skin was firmer, 97% felt their skin became more elastic, 94% felt wrinkles improved, 97% felt fine lines improved, 90% felt their overall skin condition improved, 100% felt the product was suitable for sensitive skin, 100% felt the product was gentle and non-irritating, 100% were satisfied with the overall effect / efficacy of the product, and 100% were willing to continue using the product.

[0275] Assessment Conclusion:

[0276] The evaluation results, obtained by 31 healthy Chinese female subjects with sensitive skin who used the product continuously for 28 days, showed that under the evaluation conditions, the lotion formulation of Example 3 had moisturizing, repairing, firming, and anti-wrinkle effects, and was suitable for sensitive skin and was gentle and non-irritating.

[0277] In summary:

[0278] The transdermal efficiency experiment of BSA confirms that the macromolecular transdermal microemulsion system provided by this invention can effectively achieve transdermal delivery of macromolecular substances.

[0279] Furthermore, experiments on animals using botulinum toxin have confirmed that the macromolecular transdermal microemulsion system provided by this invention has a very good transdermal effect on macromolecular substances.

[0280] Further experiments using human skin with macromolecular active ingredients have demonstrated that the macromolecular transdermal microemulsion system provided by this invention has a very good transdermal effect on macromolecular substances.

[0281] It should be noted that the above embodiments are merely illustrative examples, and the protection of this invention is not limited to the specific scope of the embodiments.

Claims

1. A macromolecular transdermal microemulsion system, characterized in that: It contains an effective amount of macromolecular active substances that can achieve the intended effect; in, The macromolecular transdermal microemulsion system comprises, by weight percentage of the total weight of the transdermal microemulsion system, the following: The oil phase content is 7-36%; The aqueous phase content is 57%-88%; The emulsion content is 8%-20%.

2. The macromolecular transdermal microemulsion system according to claim 1, characterized in that: in, The oil phase comprises 20%-30%, the aqueous phase comprises 60%-70%, and the emulsion comprises 8%-12%; and / or The transdermal microemulsion system comprises, by mass ratio: The ratio of oil phase:water phase:emulsifier is 2-2.5:6-7:

1.

3. The macromolecular transdermal microemulsion system according to claim 1 or 2, characterized in that: in, The macromolecular active substance is less than or equal to 300 kDa or less than or equal to 150 kDa. and / or The effective amount of the macromolecular active substance in the transdermal microemulsion system, calculated as a percentage of the total weight of the transdermal microemulsion system, is: The proportion of the macromolecular active substance is less than or equal to 10%, or less than or equal to 5%, or less than or equal to 2%, or less than or equal to 1%. Preferably, the macromolecular active substance is a biomolecule that can provide therapeutic, cosmetic, and skin care benefits; In another preferred embodiment, the macromolecular active substance is water-soluble.

4. The macromolecular transdermal microemulsion system according to any one of claims 1-3, characterized in that: in, The macromolecular active substances contained herein are selected from at least one or more of clostridium neurotoxin, BSA, soluble collagen, and elastin, and / or are provided by yeast extract contained in the system. Preferably, the clostridial neurotoxin is botulinum toxin of type A, B, C, D, E, F, or G, and / or the light chain of the clostridial neurotoxin contains an amino acid sequence that is at least 35%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% identical to any one of SEQ ID NO:1-7; and / or the heavy chain of the clostridial neurotoxin contains an amino acid sequence that is at least 35%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% identical to any one of SEQ ID NO:8-14.

5. The macromolecular transdermal microemulsion system according to claim 4, characterized in that: in, Based on the percentage by mass of the total weight of the transdermal microemulsion system, the system includes: The effective amount of the clostridium neurotoxin is less than or equal to 0.02%, preferably 0.00001%-0.015%, more preferably 0.0005%-0.015% or 0.0008-0.0015%; The effective amounts of soluble collagen, BSA, and elastin are all less than or equal to 5%, or less than or equal to 2%, or less than or equal to 1%. The proportion of yeast extract is 1-10%.

6. The macromolecular transdermal microemulsion system according to any one of claims 1-5, characterized in that: The system comprises, by weight percentage of the total weight of the transdermal microemulsion system, the following components: 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 deacetylated chitosan or chitosan derivative; 55-80% or 60-70% water; and the effective amount of the macromolecular active substance.

7. The macromolecular transdermal microemulsion system according to claim 6, characterized in that: in, The emulsifier is selected from any one or more of polysorbate 80, polysorbate 60, polysorbate 20, and sorbitan oleate.

8. The macromolecular transdermal microemulsion system according to any one of claims 1-7, characterized in that: It also contains preservatives. Preferably, based on a percentage of the total weight of the transdermal microemulsion system, it contains 0.1-1% preservatives by weight. Furthermore, the preservative is selected from any one or more of phenoxyethanol, methylparaben, ethylparaben, propylparaben, benzyl alcohol, and sorbic acid.

9. The macromolecular transdermal microemulsion system according to any one of claims 1-8, characterized in that: in, The average particle size of the transdermal microemulsion system is 10-15 nm.

10. The application of any one of the macromolecular transdermal microemulsion systems of claims 1-9 in achieving therapeutic, cosmetic, and skincare effects.

11. The application according to claim 10, characterized in that: When using, it can be applied transdermally through one or more of the following methods: application, transdermal instrument, patch, and spray.

12. The use of any one of the macromolecular transdermal microemulsion systems of claims 1-9 in the preparation of drugs or products that achieve therapeutic, cosmetic and skin care effects.

Images