Macromolecular transdermal microemulsion system and use thereof

Abstract

A macromolecular transdermal microemulsion system capable of enabling a macromolecular active substance to effectively transdermally exert a predetermined effect and use thereof. The macromolecular transdermal microemulsion system comprises an effective amount of a macromolecular active substance capable of exerting a predetermined effect. The macromolecular transdermal microemulsion system comprises: 7%-36% by mass of an oil phase, 57%-88% by mass of a water phase, and 8%-20% by mass of an emulsifier.

Description

A macromolecular transdermal microemulsion system and its application Technical Field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a macromolecular transdermal microemulsion system and application thereof. Background Art

[0002] The transdermal system is a method for drugs to be absorbed through the skin. It is a new way of drug administration in which drugs are absorbed through the skin into the human blood circulation and reach effective blood drug concentrations, thereby achieving disease treatment or prevention.

[0003] Transdermal drug delivery systems offer numerous advantages: they can effectively reduce pain associated with injections, effectively avoid the irritation and side effects of oral medications, effectively improve sustained-release effects, and enhance the safety of drug use and treatment. Transdermal drug delivery also offers numerous advantages, including avoiding the first-pass effect in the liver, providing more stable blood drug concentrations, and providing enhanced safety.

[0004] However, human skin mainly acts as a barrier. This structure is conducive to "internal stability and external defense", protecting the human body from external stimuli, while also preventing the entry of foreign components. If foreign components want to enter the skin, they first need to pass through the epidermis of the skin and enter the skin to take effect.

[0005] The skin consists of three parts: the epidermis, dermis, and subcutaneous tissue. The epidermis is the superficialest layer of the skin and is divided into the stratum corneum, stratum lucidum, stratum granulosum, stratum spinosum, and stratum basale. The basal layer is connected to the dermis by a basement membrane. The stratum corneum is the outermost layer of the epidermis and is composed of 5 to 15 layers of keratinocytes and intercellular lipids. This structure is figuratively likened to a "brick wall." The stratum corneum is 15 to 50 μm thick, has a very low water content (5% to 20%), and is metabolically inactive, acting as a primary barrier to transdermal absorption of chemicals. Intercellular lipids are primarily composed of 45% to 50% ceramides, 25% cholesterol, 15% long-chain free fatty acids, and 5% other lipids. The dermis, located beneath the epidermis and supporting it, is primarily composed of connective tissue, including collagen fibers, elastic fibers, and matrix. The dermis also contains other tissues, such as nerves, blood vessels, lymphatic vessels, muscles, hair follicles, sebaceous glands, and sweat glands.

[0006] Due to the dense structure of the skin's stratum corneum, most drugs cannot be absorbed into the systemic circulation, resulting in bioavailability that falls short of clinical needs. This is especially true for large-molecule bioactive drugs, which have difficulty penetrating the skin to deliver their efficacy. Therefore, improving transdermal drug penetration is a key and challenging aspect of research and development of transdermal drug delivery formulations.

[0007] Commonly used transdermal penetration enhancement technologies in physics include: microneedles, iontophoresis, electroporation technology, ultrasonic introduction technology and needle-free drug delivery system; commonly used chemical penetration enhancers in chemistry include transdermal penetration enhancers and ion pairs.

[0008] Transdermal enhancers are substances that promote transdermal absorption of drugs, a preferred method. Transdermal drug delivery systems have seen rapid development in drug development, with new products such as estradiol and testosterone entering the market. In the beauty and medical aesthetics industries, some high-end cosmetics also utilize transdermal emulsions to achieve optimal results.

[0009] However, there is still a lack of products that are good at transdermal penetration of large molecules.

[0010] Summary of the Invention

[0011] The present invention provides a macromolecular transdermal microemulsion system and its application, which can enable macromolecular active substances to effectively perform predetermined effects through the skin.

[0012] To this end, the present invention provides the following technical solutions.

[0013] The present invention provides a macromolecular transdermal microemulsion system, characterized by containing an effective amount of a macromolecular active substance capable of playing a predetermined role; wherein, in terms of mass percentage relative to the total weight of the transdermal microemulsion system, the macromolecular transdermal microemulsion system comprises: an oil phase accounting for 7-36%; a water phase accounting for 57%-88%; and an emulsion accounting for 8%-20%.

[0014] The macromolecular transdermal microemulsion system provided by the present invention also has the following characteristics: 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 is: oil phase: aqueous phase: emulsifier: 2-2.5:6-7:1 in terms of mass ratio.

[0015] 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 kD, and / or in the transdermal microemulsion system, the effective amount of the macromolecular active substance, calculated as a percentage by mass 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 biological molecule that can provide treatment, beauty and skin care; another preferred embodiment is that the macromolecular active substance is water-soluble.

[0016] The macromolecular transdermal microemulsion system provided by the present invention further has the following characteristics: wherein the macromolecular active substance contained therein is at least one or more selected from the group consisting of clostridial neurotoxin, BSA, soluble collagen and elastin, and / or is provided by the yeast extract contained in the system. Preferably, the clostridial neurotoxin is 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 that is at least 35%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 99% or 100% identical to any one of SEQ ID NOs: 1-7; and / or the heavy chain of the clostridial neurotoxin comprises 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 NOs: 8-14.

[0017] The macromolecular transdermal microemulsion system provided by the present invention also has the following characteristics: wherein, in the system, calculated as a percentage by mass of the total weight of the transdermal microemulsion system, the effective amount of the clostridial 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 the elastin are all less than or equal to 5%, or less than or equal to 2%, or less than or equal to 1%; and the yeast extract accounts for 1-10%.

[0018] The macromolecular transdermal microemulsion system provided by the present invention also has the following characteristics: the composition of the system comprises, by mass percentage of the total weight of the transdermal microemulsion system: 5%-20% or 10-20% or 13-18% of glycerol; 5%-20% or 5-15% or 8-12% of an emulsifier; 1%-10% or 3-8% of butylene glycol or propylene glycol; 0.5-5% or 1-3% of laurocapram; 0.3%-3% or 1-2% of carboxymethyl deacetylated chitosan or a chitosan derivative; 55-80% or 60-70% of 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 characteristics: 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, the preservative is contained in an amount of 0.1-1% by weight based on the total weight of the transdermal microemulsion system. Furthermore, 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 characteristics: wherein, the average particle size of the particles of the transdermal microemulsion system is 10-15 nm.

[0022] The present invention also provides an application of a macromolecular transdermal microemulsion system in achieving therapeutic, cosmetic and skin care effects.

[0023] The aforementioned application provided by the present invention is characterized in that, when used, it is applied transdermally by any one or more of smearing, transdermal instrumentation, patching, and spraying.

[0024] The present invention also provides an application of a macromolecular transdermal microemulsion system in the preparation of medicines or products for achieving therapeutic, cosmetic and skin care effects.

[0025] Functions and effects of the invention

[0026] The macromolecular transdermal microemulsion system and its application provided by the present invention, because the oil phase, water phase and emulsion in the composition of the macromolecular transdermal microemulsion system are in appropriate proportions, form an oil-in-water microemulsion with nanometer-level average particle size, and can effectively allow macromolecular active substances to be absorbed through the skin and achieve the intended effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] FIG1 is a standard curve of BSA standard according to four-parameter fitting in Example 4;

[0028] FIG2 is a diagram showing the toe-closing phenotype of the mouse experiment in Example 5. DETAILED DESCRIPTION

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

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

[0031] An emulsion is a two-phase, immiscible liquid formulation in which one phase is dispersed in the form of small droplets throughout the other phase, forming a non-uniform dispersion. The phase forming the droplets is called the dispersed phase, internal phase, or discontinuous phase, while the other phase is called the dispersion medium, external phase, or continuous phase.

[0032] Emulsions consist of an oil phase (denoted by O), an emulsifier, and a water phase (denoted by W). Depending on the type and properties of the emulsifier and the phase volume ratio (φ), they can form oil-in-water (O / W) or water-in-oil (W / O) emulsions or composite emulsions.

[0033] Aqueous phase: water or aqueous solution. Aqueous solution refers to the dissolution of water-soluble raw materials in water;

[0034] Oil phase: refers to the raw materials that are not easily soluble in water being dissolved in oily substances.

[0035] According to the size of the emulsion particles, that is, the size of the emulsion droplets, emulsions are classified into ordinary emulsions, submicron emulsions and nanoemulsions:

[0036] 1. Ordinary milk: The droplet size of ordinary milk is generally 1μm-100μm, and it is a milky white opaque liquid.

[0037] 2. Submicron emulsions: The droplet size is generally between 0.1 μm and 1.0 μm. Submicron emulsions are often used as carriers for gastrointestinal administration. Intravenous emulsions should be submicron emulsions, with particle sizes generally ranging from 0.25 μm to 0.4 μm.

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

[0039] The macromolecular transdermal microemulsion system provided by the present invention contains an effective amount of a macromolecular active substance capable of exerting a predetermined effect. The composition of the macromolecular transdermal microemulsion system, calculated by weight percentage relative to the total weight of the transdermal microemulsion system, is as follows: an oil phase accounts for 16-36% or 20-30%; an aqueous phase accounts for 55%-88% or 60-70%; and an emulsion accounts for 8%-20% or 10%-25%.

[0040] Macromolecular active substances refer to substances with larger molecular weight, mainly biological molecules.

[0041] Dalton, short for Dalton, is a commonly used unit of molecular weight. It is calculated by taking the algebraic sum of the atomic weights of all atoms in a molecule and dividing it by the number of atoms. The Dalton (Da) is numerically equal to the relative molecular mass.

[0042] In biochemistry, molecular biology, and proteomics, kDa (kilodaltons) is often used to refer to 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, or even more than one million, such as peptides, proteins, nucleic acids, polysaccharides, and antibodies.

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

[0044] Table 1 shows 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 capable of playing a predetermined role" may be oily (soluble in oil) or aqueous (soluble in water).

[0045] In one example, the macromolecular active substance of the present invention refers to a biomolecule of less than or equal to approximately 300 kDa or less than or equal to approximately 150 kDa. Preferably, the biomolecule is water-soluble, ie, exists in the aqueous phase after addition.

[0046] In one example, the predetermined effect is to achieve any one or more effects among treatment, beauty and skin care effects.

[0047] In one example, the macromolecular transdermal microemulsion system of the present invention satisfies any one or both of the requirements shown in Table 2:

[0048] As long as the macromolecular active substance can function through the system, the effective amount of the macromolecular active substance in the transdermal microemulsion system, measured as a percentage by mass of the total weight of the transdermal microemulsion system, is 10% or less, 5% or less, 2% or less, or 1% or less. Within this range, adjustments may be made based on the specific macromolecular active substance. After final adjustment, the oil phase, aqueous phase, and emulsifier in the microemulsion system as a whole meet the aforementioned ratios or proportions.

[0049] In one example, the macromolecular active substances contained are at least one or more selected from clostridial neurotoxin, soluble collagen, and elastin, and / or are provided by yeast extract contained in the system. In other words, the macromolecular active substances contained may include the following as shown in Table 3:

[0050] In one example, for any of the cases in Table 3, the contents, calculated as a percentage by mass of the total weight of the transdermal microemulsion system, are as follows:

[0051] The effective amount of the clostridial 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 clostridial neurotoxin is 10 ng / ml-10 μg / ml;

[0052] The effective amount of the soluble collagen and the elastin is less than or equal to 5%, or less than or equal to 2%, or less than or equal to 1%;

[0053] The effective amount is provided by 1-10% of the yeast extract contained in the emulsion system. For example, if the yeast extract accounts for 10% and the macromolecules contained are 5%, the effective amount is 5%.

[0054] Clostridial toxins can be used for therapeutic and cosmetic purposes. For example, botulinum toxin specifically binds to presynaptic receptors on peripheral cholinergic nerve endings. By cleaving the synaptic-associated protein SNAP-25, it interferes with the exocytosis of presynaptic vesicles, inhibiting the release of acetylcholine (ACh) from nerve endings. This leads to chemical denervation of muscles, resulting in muscle relaxation and relief of muscle spasms. This muscle chemical denervation effect of botulinum toxin is the theoretical basis for its use in regulating hypertonia, movement disorders, and dynamic wrinkles. Furthermore, multiple injections into target muscles can lead to disuse atrophy and reduction in muscle size, leading to its use in body contouring. Currently, botulinum toxin is used in the treatment of hemifacial spasm, essential blepharospasm, spastic torticollis, spastic cerebral palsy, post-stroke limb spasticity, tremor, and other dystonias. The application of botulinum toxin in the field of minimally invasive plastic surgery includes the removal of various dynamic wrinkles such as frown lines, forehead lines, and crow's feet; the beautification of facial contours such as eyebrow height adjustment, mental muscle relaxation, mandibular margin lifting, and neck cord injection; and the shaping of body contours such as masseter muscle reduction, gastrocnemius muscle reduction, and trapezius muscle reduction.

[0055] The clostridial toxins herein refer to natural clostridial toxins and toxins having similar structures and functions to natural clostridial toxins.

[0056] Despite differences in amino acid sequences and immunogenicity among various toxin types, they all exhibit similar molecular structures. Clostridial neurotoxins are produced by toxigenic Clostridium as non-toxic single-chain polypeptides of approximately 150 kD. They become active only after being cleaved by bacterial proteases or in vitro proteases into a two-chain form, consisting of a light chain (L chain, amino terminus of the toxin, 50 kD) and a heavy chain (H chain, carboxyl terminus of the toxin, 100 kD) linked by a disulfide bond. The heavy chain is composed of two domains: the Hn (amino terminus, 50 kD) and the Hc (carboxyl terminus, 50 kD).

[0057] 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 that is at least 35%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 99%, or 100% identical to any one of SEQ ID NOs: 1-7; and / or the heavy chain of the clostridial neurotoxin comprises 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 NOs: 8-14. The sequences are shown in Table 4.

[0058] Botulinum toxin-related products have broad application prospects in the treatment of clinical spastic disorders, glandular hypersecretion, neuropathic pain, and cosmetic wrinkle reduction. However, due to their large molecular size, these products are currently injected, making it difficult for them to directly penetrate the skin and reach their target site.

[0059] Currently, only botulinum toxin types A and B are available for pharmaceutical use, but the vast majority of botulinum toxins used in medicine are type A. Type A botulinum toxins currently in clinical use include Botox (Allergan, USA), Dysport (Speywood, UK), and Hengli (BTX-A) (Lanzhou Institute of Biological Products, my country). Botulinum toxin-related products have broad application prospects in the treatment of clinical spastic disorders, glandular hypersecretion, neuropathic pain, and cosmetic wrinkle reduction. However, these products are currently administered via injection. Our macromolecular transdermal microemulsion system, however, has demonstrated in vivo efficacy studies in mice that it allows biological macromolecules, such as botulinum toxin, to achieve their intended effects through transdermal delivery.

[0060] Yeast extract (also known as yeast flavor, abbreviated as YE) is a pure natural product that comes in a brownish-yellow soluble paste or light yellow powder, made from protein-rich edible yeast according to the Chinese Pharmacopoeia. It is refined using modern biotechnology, including autolysis, enzymatic hydrolysis, separation, and concentration, to degrade the proteins and nucleic acids within yeast cells. Yeast extract's main components include amino acids and nucleotides, as well as monosaccharides, polysaccharides, minerals, and vitamins.

[0061] The main application effects of yeast extract are skin rejuvenation, anti-aging and water retention. Yeast extract has a good skin rejuvenation effect, which is achieved through three main modes of action: (1) reducing the transfer of existing melanin to surface cells; (2) promoting the metabolism of melanin-containing keratinocytes to accelerate keratin shedding; (3) promoting the synthesis of epidermal proteins to regulate skin texture. Yeast extract has a good free radical scavenging and anti-aging effect. It can regulate skin texture, fine lines and wrinkles, increase skin elasticity, make the skin smooth and delicate, and can also reduce and prevent the dull skin color caused by the early aging process. Yeast extract also has a strong water retention effect: rich in natural moisturizing factors, it can promote the stratum corneum to retain moisture more effectively, give the skin deep moisture and prevent dryness; by increasing the content of ceramide in the stratum corneum, it can strengthen the skin barrier function and reduce transepidermal water loss; it can also repair the damaged stratum corneum lipid barrier, improve skin resistance and regulate skin condition.

[0062] Soluble collagen is a naturally occurring protein that performs important functions in the skin. While collagen maintains skin's elasticity and firmness, it decreases with age, leading to sagging and wrinkles. Soluble collagen, as a form that penetrates deeper into the skin, can replenish collagen and improve its condition.

[0063] Soluble collagen can stimulate and promote the synthesis of collagen, help repair damaged collagen, and promote the production of new collagen, thereby improving the elasticity and firmness of the skin and improving wrinkles and fine lines.

[0064] Elastin is a key component of the skin and the foundation of its youthfulness. Elastin peptides promote skin damage repair, blood vessel regeneration, and fibroblast proliferation. They also enhance skin elasticity, resulting in smooth, resilient skin.

[0065] In one example, the composition of the macromolecular transdermal microemulsion system of the present invention is shown in Table 5:

[0066] Preferably, the example of Table 5 is composed of Table 6:

[0067] Chitosan, a natural alkaline polysaccharide, possesses film-forming, antibacterial, anti-inflammatory, hemostatic, analgesic, antipruritic, scar-removing, wound-healing, and antioxidant properties. Chitosan has a strong inhibitory effect on bacteria, yeast, fungi, and other microorganisms. It has significant inhibitory effects on common bacteria found on the human epidermis, such as Staphylococcus epidermidis, Escherichia coli, and Candida albicans tropicalis, as well as Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus aureus pyogenes, common in burn patients. Chitosan possesses potent moisturizing properties, even exceeding those of sodium hyaluronate, earning it the nickname "super moisturizer." Chitosan can absorb heavy metals and cosmetic residues. Its restorative properties can repair the skin barrier and are beneficial for sensitive and acne-prone skin.

[0068] In one example, the macromolecular transdermal microemulsion system further contains a preservative. Preferably, the preservative is present in an amount of 0.1-1% by weight relative to the total weight of the transdermal microemulsion system. Furthermore, the preservative is selected from any one or more of phenoxyethanol, methylparaben, ethylparaben, propylparaben, benzyl alcohol, and sorbic acid.

[0069] The average particle size of the particles of the macromolecular transdermal microemulsion system of the present invention is nanometer-scale, preferably, 10-15 nm on average.

[0070] Table 5 is a specific example of the composition of the macromolecular transdermal microemulsion system provided by the present invention.

[0071] The macromolecular transdermal microemulsion system of the present invention is used to achieve therapeutic, cosmetic and skin care effects. When used, it can be transdermally applied by any one or more of smearing, transdermal instruments, attachment and spraying.

[0072] The macromolecular transdermal microemulsion system of the present invention can also be used to prepare medicines or products for achieving therapeutic, cosmetic and skin care effects.

[0073] In addition, in the present invention, water is preferably any one or more combinations of distilled water, drinking water, sterile water, ultrapure water and deionized water.

[0074] In the following examples, some of the components involved are described as follows:

[0075] The main applications of superoxide dismutase in cosmetics include: (1) as a cosmetic additive to prevent skin aging and provide skin care benefits; (2) prevention and treatment of related skin diseases. These applications have been widely used abroad. Currently, many high-end cosmetics at home and abroad have added SOD and made it into facial masks, lotions, and creams.

[0076] Vitamins are essential organic substances for the human body. Vitamin A promotes epidermal cell proliferation and increases the formation of dermal collagen and elastin, offering anti-aging benefits. Vitamin E is a natural antioxidant that boosts metabolism and improves skin elasticity. Vitamin C has strong antioxidant properties, promoting collagen synthesis and inhibiting collagen breakdown. Vitamins C and E synergistically scavenge free radicals.

[0077] In addition to essential vitamins, a variety of natural plant active ingredients have been shown to have anti-wrinkle and firming functions, such as Centella asiatica extract, Osmanthus fragrans extract, and Calendula officinalis extract. These raw materials are currently widely used in anti-wrinkle and firming cosmetics.

[0078] Example 1

[0079] The formula of the macromolecular transdermal microemulsion system provided in this embodiment is shown in Table 6:

[0080] Preparation method:

[0081] (1) Using an electronic balance, weigh purified water, carboxymethyl chitosan, and botulinum toxin A according to the formula into a 3 L beaker 1 and stir at room temperature to dissolve (speed: 400-500 rpm).

[0082] (2) Using an electronic balance, weigh polysorbate 80, laurocapram, phenoxyethanol, propylene glycol, and glycerin according to the formula amount into a 2 L beaker 2, and stir evenly at room temperature (speed: 250-300 rpm).

[0083] (3) Add the solution in beaker 2 to beaker 1 and stir evenly at room temperature (speed: 400-500 rpm).

[0084] (4) Add purified water to the solution prepared in step 3) to make up to 1000 g, and stir at room temperature to mix.

[0085] Example 2

[0086] The formula of the macromolecular transdermal microemulsion system provided in this example is shown in Table 7.

[0087] Preparation method:

[0088] (1) Using an electronic balance, weigh purified water, carboxymethyl chitosan, yeast extract, superoxide dismutase (SOD), and sodium ascorbyl phosphate according to the formula into a 3 L beaker 1, and stir at room temperature (speed: 400-500 rpm) to dissolve.

[0089] (2) Using an electronic balance, weigh polysorbate 80, tocopherol (vitamin E), retinyl palmitate, phenoxyethanol, (daily use) fragrance, butylene glycol and glycerin according to the formula amount into a 2L beaker 2, and stir evenly at room temperature (speed: 250-300 rpm).

[0090] (3) Add the solution in beaker 2 to beaker 1 and stir evenly at room temperature (speed: 400-500 rpm).

[0091] (4) Add purified water to the solution prepared in step (3) and make up to room temperature.

[0092] Stir to mix.

[0093] Example 3

[0094] The formula of the macromolecular transdermal microemulsion system provided in this example is shown in Table 8.

[0095] Preparation method:

[0096] (1) Using an electronic balance, weigh purified water, hydroxypropyl tetrahydropyrantriol, carnosine, sodium polyglutamate, biotin, Algae extract, Calendula officinalis extract, Centella asiatica extract, Aloe vera extract, carboxymethyl chitosan, sodium ascorbyl phosphate, yeast extract, superoxide dismutase (SOD), soluble collagen, and elastin according to the formula amount into a 3 L beaker 1, and stir at room temperature to dissolve (speed: 400-500 rpm).

[0097] (2) Using an electronic balance, weigh polysorbate 80, tocopherol (vitamin E), retinyl palmitate, phenoxyethanol, (daily use) fragrance, butylene glycol and glycerin according to the formula amount into a 2L beaker 2, and stir evenly at room temperature (speed: 250-300 rpm).

[0098] (3) Add the solution in beaker 2 to beaker 1 and stir evenly at room temperature (speed: 400-500 rpm).

[0099] (4) Add purified water to the solution prepared in step (3) to make up to 1000 g, and stir at room temperature to mix.

[0100] The preparations obtained in Examples 1-3 have good emulsification effect, high transparency, and no stratification. The average particle size detected by a nanoparticle size potentiometer is 10-15 nm. In particular, the preparation obtained in Example 3 has an average particle size of 12.99 nm.

[0101] Comparative Example 1

[0102] The formula of this comparative example is shown in Table 9

[0103] Preparation method:

[0104] (1) Add the acid ester to a 2L beaker and stir for 5 minutes; weigh oleyl alcohol and oleic acid according to the formula using an electronic balance and add them to the beaker and stir for 5 minutes; weigh laurocapram and phenoxyethanol according to the formula using an electronic balance and add them to the beaker and stir evenly at room temperature (speed: 200-250 rpm, 5 minutes).

[0105] (2) Use an electronic balance to weigh propylene glycol and menthol according to the formula and add them to a blue-capped bottle in sequence. Heat to 60°C (about 10 minutes) to fully dissolve them, then add them to the solution obtained in step 1) and stir evenly at room temperature (speed: 200-250 rpm, 5 minutes).

[0106] (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).

[0107] (4) Add purified water to the solution prepared in step (3) to make up to 1000 g, and stir at room temperature to mix.

[0108] Comparative Examples 2-10

[0109] The formulas of Comparative Examples 2-10 are shown in Table 10, calculated by mass percentage:

[0110] According to the respective formulations in Comparative Examples 2-10, the oil phase is fully mixed and dissolved, the water phase is fully mixed and dissolved, and then the phases are mixed to obtain the products. The specific preparation methods are all conventional techniques and can be selected by referring to the previous embodiments, so they will not be described one by one.

[0111] The appearance of the prepared comparative examples 2-10 was not as good as that of the examples 1-3, as shown in Table 11:

[0112] Example 4

[0113] This experiment was used to detect the transdermal efficiency.

[0114] The botulinum toxin A added in Example 1 and Comparative Example 1 was replaced with BSA protein. The concentration of the BSA protein emulsion prepared in Example 1 was 1.0 mg / ml, and the concentration of the BSA protein emulsion prepared in Comparative Example 1 was 0.62 mg / ml. Transdermal experiments were conducted using the Franz diffusion cell method.

[0115] Test equipment and materials

[0116] 1. Reagents

[0117] Phosphate buffer, protein standards, test products, protein detection kit (ultra-high sensitivity);

[0118] 2. Consumables

[0119] For transdermal evaluation, isolated pig skin, centrifuge tubes, EP tubes, pipette tips, and pre-coated microtiter plates were used;

[0120] 3. Equipment

[0121] TP-6 transdermal diffusion instrument (Tianjin Jingtuo Instrument Technology Co., Ltd.), multifunctional microplate reader (Thermo scientific, Varioskan Lux), medical refrigerator (Qingdao Haier Biomedical Co., Ltd.), biochemical incubator (Shanghai Yiheng Scientific Instrument Co., Ltd.), and pipette (Eppendorf AG).

[0122] 1. Preparation before the experiment:

[0123] (1) The pig skin required for the experiment was taken out of the -20℃ refrigerator and thawed at 4℃, then washed with physiological saline and set aside.

[0124] (2) Before the experiment, clean the supply chamber, diffusion chamber (receiving chamber) and stirring bar.

[0125] (3) Add pure water to the water tank, preferably flush with the diffusion cell frame, and set the temperature to 32℃ and the speed.

[0126] (4) After the temperature of the TP-6 ​​transdermal diffusion instrument reaches the set temperature, place the pig skin between the supply chamber and the diffusion chamber. Stir at a constant speed to maintain the solution at an isothermal state and uniform concentration of the exudate.

[0127] 2. Experimental methods:

[0128] (1) Set up three experimental groups and one blank group.

[0129] (2) Adding BSA protein test solution: 150 μl (1 mg / ml) of the BSA protein emulsion test solution prepared in Example 1, 150 μl (0.62 mg / ml) of the BSA protein emulsion test solution prepared in Comparative Example 1, and 150 μl (1 mg / ml) of the BSA protein test solution dissolved in PBS were added to the first three supply chambers, respectively. 150 μl of PBS was added to the blank group.

[0130] (3) Fill the receiving chamber with PBS. Take four 1.5 ml EP tubes and sample 400 μl of the receiving chamber liquid from the sampling port every hour for subsequent OD measurement. Mark the sampling time and experimental group number. Sampling should be performed for a total of 6 hours. After sampling, each receiving chamber must be replenished with 400 μl of PBS to maintain the receiving chamber volume.

[0131] (4) All collected samples were placed in a 4°C refrigerator for later use.

[0132] 3. ELISA test:

[0133] 1. Reagent preparation:

[0134] a. Before use, equilibrate all reagents to room temperature (18-25°C). Set the detection wavelength to 450 nm according to the instructions of the microplate reader and preheat the plate for 15 minutes before reading.

[0135] b. Wash Buffer: Dilute 20 ml of concentrated wash buffer with 580 ml of deionized water to prepare 600 ml of wash buffer.

[0136] c. Standard Working Solution: First, centrifuge the standard at 1000g for 1 minute. Add 2ml of Standard Sample Diluent and mix thoroughly. Pipette 300μl of the 20,000ng / ml Standard Diluent into the first tube and mix thoroughly to create the 6,666.7ng / ml working solution. Transfer another 300μl of this solution from the first tube to the second tube. Mix each tube thoroughly before transferring the solution to the next tube. Set up six dilutions of the standard: 20,000ng / ml, 6,666.7ng / ml, 2,222.2ng / ml, 740.7ng / ml, 246.9ng / ml, and 0ng / ml.

[0137] 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. Briefly centrifuge the stock solution tube before use. Dilute the 100× concentrated Detection Reagent A to 1× working solution with Detection Diluent A (e.g., 10 μl Detection Reagent A + 990 μl Detection Reagent A Diluent).

[0138] e. Detection Reagent B Working Solution: Calculate the required volume (100 μl / well) before the experiment. Prepare 100-200 μl more than the calculated volume. Briefly centrifuge the stock solution tube before use. Dilute the 100× concentrated Detection Reagent B to 1× working solution with Detection Diluent B (e.g., 10 μl Detection Reagent B + 990 μl Detection Reagent B Diluent).

[0139] 2. Experimental Methods

[0140] (1) Set up standard wells, sample wells, and blank wells. Add six standards of different concentrations (including zero well, 50 μL / well) in sequence, add 50 μL of the sample to be tested (samples collected from the sampling port of the receiving chamber) to the other wells, and then immediately add the detection reagent A working solution, 50 μL / well, gently shake to mix, cover with the sealing film provided by the kit, and incubate at 37°C for 60 minutes.

[0141] (2) Discard all liquid from the wells and add 350 μL of wash buffer to each well. Let the wells soak for 60 seconds. Pour out the liquid from each well and pat dry on clean absorbent paper. Repeat this washing step three times.

[0142] (3) Add 100 μL of detection reagent B working solution to each well, cover with sealing film, and incubate at 37°C

[0143] 30 minutes.

[0144] (4) Discard the liquid in each well and repeat the washing process in step 2 5 times.

[0145] (5) Add 90 μL of TMB reagent to each well, cover with a new sealing film, and incubate at 37°C in the dark.

[0146] 10-20 minutes.

[0147] (6) Add 50 μL of stop solution to each well in the same order as the color development solution, and gently shake to mix.

[0148] (7) Ensure that there is no air bubble or water mist at the bottom of the ELISA plate wells, and immediately measure the absorbance of each well at 450nm

[0149] Determine the OD value and record the reading of the microplate reader.

[0150] 4. Test Data and Conclusions

[0151] (1) The standard curve of BSA standard according to four-parameter fitting is shown in Figure 1.

[0152] Standard curve drawing:

[0153] Process the OD value data of the detected standard protein:

[0154] The data were fitted with four parameters to obtain the standard curve. The four-parameter fitting formula is:

[0155] The specific standard curve is shown in Figure 1.

[0156] (2) Sample test results

[0157] Table 12 shows the BSA concentrations permeating through the skin of different test samples at different time periods calculated based on the standard curve.

[0158] Based on the results, perform transdermal calculation:

[0159] (1) After 24 hours, the BSA emulsion protein test solution prepared in Example 1 had a concentration of 239.6 ng / ml in the receptor compartment, which had a volume of 15 ml, while the BSA concentration in the donor compartment was 1 mg / ml, and the volume added to the donor compartment was 150 μl. Based on this, the transdermal efficiency of the BSA emulsion prepared in Example 1 was calculated to be 2.40%.

[0160] (2) Comparative Example 2 prepared a BSA emulsion protein test solution. After 24 hours, the concentration in the receiving chamber was 147.1 ng / ml, and its volume was 15 ml. The BSA concentration in the donor pool was 0.62 mg / ml, and the volume added to the donor chamber was 150 μl. Based on this, the transdermal efficiency of the BSA emulsion prepared in Example 5 was calculated to be 2.37%.

[0161] (3) After 24 hours, the BSA protein test solution dissolved in PBS has a concentration of 125.4 ng / ml in the receiving chamber, which has a volume of 15 ml, while the BSA concentration in the donor chamber is 1 mg / ml, and the volume added to the donor chamber is 150 ul. Based on this, the transdermal efficiency of PBS is calculated to be 1.25%.

[0162] It can be seen that, compared with the control, the transdermal effects of Example 1 and Comparative Example 2 are very good, and the transdermal effect of Example 1 is better than that of Comparative Example 2.

[0163] Example 5

[0164] The efficacy of botulinum toxin A microemulsions prepared in Example 1 and Comparative Example 1 at a concentration of 10 μg / ml was verified in mice.

[0165] To compare the transdermal efficiency of the macromolecular active substance between Example 1 and Comparative Example 1, we used 17-19 g SPF CD-1 (ICR) mice. After anesthesia with tribromoethanol, the skin on the right hind leg was shaved. Before each application, the application area was cleaned with a medical cotton swab dipped in 10% alcohol and allowed to dry before application. 200 μl of botulinum toxin A microemulsion was applied to the skin of the right hind leg of the mice once daily for 7 consecutive days. The left hind leg of the mice was not treated with botulinum toxin A.

[0166] The experimental results showed that 2-3 days after the mice were treated with botulinum toxin A, the toes of the right hind legs showed a phenotype of closure (2-5 toes closed together). The experimental results of Example 1 and Comparative Example 1 are shown in Table 13. The emulsion prepared in the formula of Example 1 showed syndactyly on D2: 33% of the experimental mice had two toes closed, and 33% of the experimental mice had five toes closed; on D3, syndactyly: 33% of the experimental mice had two toes closed, and 33% of the experimental mice had five toes closed; on D4, syndactyly: 66% of the experimental mice had three toes closed, and 33% of the experimental mice had five toes closed; on D5, syndactyly: 100% of the experimental mice had five toes closed.

[0167] The emulsion prepared in the formula of Comparative Example 1 showed no syndactyly on D2; syndactyly on D3: 33% of the experimental mice had two toes syndactyly; syndactyly on D4: 66% of the experimental mice had two toes syndactyly, and 33% of the experimental mice had three toes syndactyly; syndactyly on D5: 33% of the experimental mice had three toes syndactyly, and 66% of the experimental mice had four toes syndactyly.

[0168] The results in Table 13 show that the botulinum toxin A microemulsion prepared in Example 1 had significantly better efficacy in mice than the microemulsion in Comparative Example 1. The microemulsion in Example 1 not only took effect earlier but also had better efficacy than the microemulsion in Example 1. The phenotypes of toe-pushing in Example 1 and Comparative Example 1 are shown in Figure 2.

[0169] According to literature reports, the toes of mice will appear to be folded together after injection of botulinum toxin A into the gastrocnemius muscle.

[0170] It can be seen that we used the emulsion delivery system we developed to make botulinum toxin A into a microemulsion and applied it to the skin of mice, which also achieved the syndactyly phenotype after the injection of botulinum toxin A. This proved in mice that the emulsion system we developed is more effective.

[0171] Example 7

[0172] The efficacy of the anti-wrinkle and firming cosmetic microemulsion prepared in Example 3 was tested on the human body.

[0173] 1. Under normal circumstances, adult subjects 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 mild and non-irritating.

[0174] 2. Subjects: A total of 31 healthy Chinese women with sensitive skin (screened using the SGS (SGS Standards Technical Services Co., Ltd.) internal sensitive skin questionnaire) were evaluated. They ranged in age from 31 to 60 years (mean age 52.65 ± 6.08 years) and met the voluntary inclusion and exclusion criteria. The face was the evaluation area.

[0175] Subject inclusion and exclusion criteria:

[0176] The subjects for this evaluation were screened from the CPCH efficacy laboratory subject information database of SGS, and healthy subjects who met the following inclusion and exclusion criteria were selected.

[0177] (1) Selection criteria

[0178] Healthy women, age: 28-60 years old;

[0179] Ethnicity: Asian (Chinese);

[0180] The subjects had sensitive facial skin (screened by the SGS internal sensitive skin questionnaire);

[0181] The subjects rated their facial discomfort symptoms as (non-persistent) itching or tingling as 4-7 points (0-9 scale, self-assessment questionnaire);

[0182] The subjects perceived their facial skin as dull, sagging, and lacking in elasticity;

[0183] Forehead wrinkle visual score 3-6 (according to SGS internal atlas);

[0184] Wrinkles on the outer canthus of the eye on one random side were visually scored as 3-6 (according to the SGS internal atlas);

[0185] The visual score of the nasolabial wrinkles (nasolabial folds) on one side is 3-6 (according to the SGS internal atlas);

[0186] Transepidermal water loss (TEWL) > 15 g / h / m2 on one cheek area;

[0187] The stratum corneum moisture content of one cheek area was measured to be <60 a.u.;

[0188] The facial skin has no obvious skin lesions, scars, hair, etc.;

[0189] Can cooperate well with the evaluation project according to the requirements of the plan and maintain regular life during the study period.

[0190] (2) Exclusion criteria

[0191] Anyone with any of the following conditions must be excluded from this study:

[0192] Those who do not agree to sign the informed consent form;

[0193] Those who are unwilling to comply with the requirements of the program;

[0194] concurrently participate in any other clinical research study;

[0195] Evaluate the use of cosmetics and / or medicines on the day;

[0196] She reported being pregnant and breastfeeding;

[0197] were receiving medication during the study period;

[0198] Subjects with infectious skin diseases or atopic dermatitis;

[0199] Those with skin abnormalities such as moles, capillary dilation, etc. at the assessment site;

[0200] Subjects who had received skin peeling or skin treatment within 3 months before participating in the assessment;

[0201] Subjects who had received immunosuppressant therapy within 3 months before participating in the assessment;

[0202] Subjects who received systemic steroid treatment or phototherapy within 1 month before participating in the assessment;

[0203] Use of topical medications and / or special-effect cosmetics (claiming moisturizing, repairing, firming, or anti-wrinkle) on the affected area within 2 weeks prior to the assessment;

[0204] There are lesions, obvious marks, or other abnormalities in the evaluation area, making measurement difficult;

[0205] Subjects with severe reactions or allergies to cosmetics, medications, or general light exposure;

[0206] In addition to the above matters, when the project leader determines that it is not appropriate to conduct the evaluation.

[0207] 3. Directions: After cleansing, thoroughly remove oil and keep skin dry. Take two drops of the essence (microemulsion) from the dropper and apply to wrinkled areas such as the corners of the eyes, forehead, nasolabial folds, and chin. Massage until the essence (microemulsion) is absorbed. It is recommended to use daily, seven days a week. A four-week usage cycle is recommended. (When not in use, please store the product in a refrigerator at a constant temperature of 2-8°C.)

[0208] 4. Evaluation period: before using the product (D0), 14 days after using the product (D14), and 28 days after using the product (D28).

[0209] 5. Evaluation parameters:

[0210] (1) Image acquisition

[0211] Primos CR is used to collect facial images and analyze skin wrinkles. A decrease in the analysis value indicates improvement in skin wrinkles.

[0212] (2) Skin elasticity

[0213] Skin elasticity tester The MPA580 is used to test skin elasticity. Increased values ​​for R2, R5, and R7 indicate improved skin elasticity.

[0214] (3) Skin firming

[0215] Skin elasticity tester The MPA580 is used to measure skin firmness. A decrease in the F4 measurement value indicates an improvement in skin firmness.

[0216] (4) Water content of the skin stratum corneum

[0217] Skin moisture content meter The CM825 is used to measure the moisture content of the skin's stratum corneum. A larger measurement value indicates an increase in moisture content in the skin's stratum corneum.

[0218] (5) Transepidermal water loss rate

[0219] Skin water loss meter TM Hex is used to measure the transepidermal water loss rate of the skin. A smaller measurement value indicates an improvement in the skin barrier.

[0220] (6)TC value

[0221] The TC value is a secondary parameter in non-invasive testing, which indicates the amount of water lost through the epidermis per unit area per unit time. A smaller analysis value indicates that the skin barrier function has improved.

[0222] (7) Skin moisture distribution map

[0223] The MoistureMap MM 200 is a unique instrument that uses electrical capacitance imaging to visualize hydration distribution and texture characteristics. MGL represents the mean grayscale value of skin moisture distribution, with lower values ​​indicating higher hydration.

[0224] (8) Skin glossiness

[0225] Skin gloss meter GL200 is used to measure skin glossiness. A larger measurement value indicates an increase in skin glossiness.

[0226] (9) Subject self-assessment

[0227] The subjects conducted self-assessments based on their own usage.

[0228] 6. Evaluation plan design:

[0229] (1) Before using the product (D0):

[0230] After arriving at SGS, the subjects cleansed their face with a cleansing product and dried their skin with a dry, dandruff-free facial tissue. They sat in a laboratory at a temperature of 21±1°C and 50±10% RH for 30 minutes and were visually evaluated by a dermatologist. Those who met the inclusion criteria proceeded to the next step of the evaluation.

[0231] Laboratory technicians measured facial skin stratum corneum moisture content (Corneometer) and transepidermal water loss (Tewameter), and those who met the inclusion criteria entered the next step of evaluation;

[0232] Laboratory technicians collected Primos CR images, Moisture Map MM200, Cutometer, and Glossymeter measurements from eligible subjects.

[0233] (2) Subjects fill out questionnaires

[0234] The laboratory technicians explain how to use the product to the subjects and distribute the product. The subjects try the samples on site under the supervision of the laboratory technicians. If any adverse reactions occur, they need to fill in the usage log in time.

[0235] (3) 14 days after using the product (D14)

[0236] Upon arrival at SGS, participants cleansed their faces with cleansing products and dried their skin with dry, dandruff-free facial tissue. They sat in a laboratory at 21±1°C and 50±5% relative humidity for 30 minutes. Laboratory technicians collected Primos CR images of the participants' faces and measured their skin using the Cutometer, Moisture Map MM200, Tewameter, Corneometer, and Glossymeter instruments.

[0237] Laboratory technicians weigh products and check product usage logs

[0238] The subject leaves SGS.

[0239] (4) 28 days after using the product (D28)

[0240] Upon arrival at SGS, participants cleansed their faces with cleansing products and dried their skin with dry, dandruff-free facial tissue. They sat in a laboratory at 21±1°C and 50±5% relative humidity for 30 minutes. Laboratory technicians collected Primos CR images of the participants' faces and measured their skin using the Cutometer, Moisture Map MM200, Tewameter, Corneometer, and Glossymeter instruments.

[0241] The subjects filled out the questionnaire;

[0242] Laboratory technicians weigh and recover products and check and recover product usage logs;

[0243] The subject leaves SGS.

[0244] (5) Data statistics: SPSS 28.0 was used to analyze the data. A normal distribution test was performed on the evaluation data. If the evaluation data were normally distributed, the T test was used for statistical analysis. If the evaluation data were non-normally distributed, the rank sum test was used for statistical analysis. Rank data were statistically analyzed using the rank sum test. The statistical significance level was P < 0.05.

[0245] The product was used continuously for 28 days by 31 healthy Chinese female subjects with sensitive skin. The evaluation results showed that under the evaluation conditions, the product had moisturizing, repairing, firming, and anti-wrinkle effects after 14 days, and was suitable for sensitive skin and mild and non-irritating. The specific results are as follows

[0246] (1) Instrument evaluation results (Table 14)

[0247] (2) Subjects’ self-assessment (satisfaction)

[0248] After using the product for 14 days, 94% of the subjects felt that their skin was deeply hydrated, 94% of the subjects felt that their skin was hydrated and plump, 94% of the subjects felt that the moisturizing effect was good, 90% of the subjects felt that their skin was firmer, 87% of the subjects felt that their skin became more elastic, 87% of the subjects felt that wrinkles were improved, 87% of the subjects felt that fine lines were improved, 87% of the subjects felt that the overall condition of the skin was improved, 90% of the subjects felt that the product was suitable for sensitive skin, and 100% of the subjects felt that the product was gentle and non-irritating.

[0249] After using the product for 28 days, 100% of the subjects felt that it was deeply hydrated, 100% of the subjects felt that their skin was hydrated and plump, 100% of the subjects felt that the moisturizing effect was good, 100% of the subjects felt that their skin was firmer, 97% of the subjects felt that their skin became elastic, 94% of the subjects felt that wrinkles were improved, 97% of the subjects felt that fine lines were improved, 90% of the subjects felt that the overall condition of the skin was improved, 100% of the subjects felt that the product was suitable for sensitive skin, 100% of the subjects felt that the product was gentle and non-irritating, 100% of the subjects were satisfied with the overall effect / efficacy of the product, and 100% of the subjects were willing to continue using the product.

[0250] Evaluation conclusion:

[0251] The product was used continuously for 28 days by 31 healthy Chinese female subjects with sensitive skin. The evaluation results showed that under the evaluation conditions, the emulsion preparation of Example 3 has the effects of moisturizing, repairing, firming and anti-wrinkle, and is suitable for sensitive skin and is mild and non-irritating.

[0252] In summary:

[0253] The transdermal efficiency experiment of BSA confirmed that the macromolecular transdermal microemulsion system provided by the present invention can effectively achieve the transdermal delivery of macromolecular substances.

[0254] Furthermore, experiments using botulinum toxin on animals confirmed that the macromolecular transdermal microemulsion system provided by the present invention has a very good transdermal effect on macromolecular substances.

[0255] Experiments using macromolecular active ingredients on human skin also demonstrate that the macromolecular transdermal microemulsion system provided by the present invention has a very good transdermal effect on macromolecular substances.

[0256] It should be noted that the above embodiments are merely examples for specific description, and the protection of the present invention is not limited to the specific scope of the embodiments.

Claims

1. A macromolecular transdermal microemulsion system, characterized in that: Contains an effective amount of macromolecular active substances that can play a predetermined role; in, Calculated by mass percentage of the total weight of the transdermal microemulsion system, the composition of the macromolecular transdermal microemulsion system is: The oil phase accounts for 7-36%; The proportion of water phase is: 57%-88%; The proportion of emulsion is: 8%-20%.

2. The macromolecular transdermal microemulsion system according to claim 1, characterized in that: in, The oil phase accounts for 20%-30%, the water phase accounts for 60%-70%, and the emulsion accounts for 8%-12%; and / or According to the mass ratio, the composition of the transdermal microemulsion system is: 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 Calculated by 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 biological molecule that can provide treatment, beauty and skin care; Another preferred embodiment is that the macromolecular active substance is water-soluble.

4. The macromolecular transdermal microemulsion system according to any one of claims 1 to 3, characterized in that: in, The macromolecular active substances contained are at least selected from any one or more of clostridial neurotoxin, BSA, soluble collagen and elastin, and / or are provided by the yeast extract contained in the system, Preferably, 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 that is at least 35%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 99% or 100% identical to any one of SEQ ID NOs: 1-7; and / or the heavy chain of the clostridial neurotoxin comprises 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 NOs: 8-14.

5. The macromolecular transdermal microemulsion system according to claim 4, characterized in that: in, Measured by weight percentage of the total weight of the transdermal microemulsion system, in the system: The effective amount of the clostridial 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 the 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 to 5, characterized in that: Measured by mass percentage of the total weight of the transdermal microemulsion system, the system comprises: 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 to 7, characterized in that: It also contains preservatives. Preferably, the transdermal microemulsion system contains 0.1-1% by weight of a preservative, 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 to 8, characterized in that: in, The average particle size of the particles in the transdermal microemulsion system is 10-15 nm.

10. Use of any one of the macromolecular transdermal microemulsion systems of claims 1 to 9 in achieving therapeutic, cosmetic and skin care effects.

11. The use according to claim 10, characterized in that: When in use, it is applied transdermally by any one or more of the following methods: applying, transdermal instrument, patching and spraying.

12. Use of any one of the macromolecular transdermal microemulsion systems of claims 1 to 9 in the preparation of drugs or products for achieving therapeutic, cosmetic and skin care effects.

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