Baicalin-cyclodextrin-vesicle compound and application thereof

Abstract

The invention relates to a baicalin-cyclodextrin-vesicle compound as well as a preparation method and application thereof. Specifically, the invention discloses a baicalin-cyclodextrin-vesicle compound, the baicalin-cyclodextrin-vesicle compound comprises (i) an active component, and the active component comprises baicalin; (ii) a cyclodextrin; and (iii) a vesicle material, the vesicle material comprising vesicles of microbial origin; in addition, the cyclodextrin (ii) comprises the active ingredient (i) to form nanoparticles; and the (iii) vesicle material wraps the nanoparticles to form a baicalin-cyclodextrin-vesicle compound. The baicalin-cyclodextrin-vesicle compound has the performance of high transdermal efficiency, high bioavailability and high drug loading capacity, has the functions of aging resistance, oxidation resistance, whitening, inflammation resistance and the like, and can be applied to cosmetics.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and in particular to a baicalin-cyclodextrin-vesicle complex and an application thereof. Background Art

[0002] In cosmetic applications, the occurrence of oxidative aging-related problems is mainly concentrated in the dermis of the skin, so whether the raw materials can be delivered to the dermis and the raw material intake of dermal cells is crucial for the antioxidant and anti-aging effects. Baicalin can scavenge oxygen free radicals and inhibit the production of melanin, and can be used in medicine and cosmetics. However, as a water- and oil-insoluble ingredient, baicalin is very difficult to use in formulas in its original form. It can only be used in cosmetics in extremely small amounts and cannot reach the effective concentration. In addition, baicalin in its original form is not only difficult to reach the dermis, but also has the problem of easy discoloration.

[0003] At present, liposome encapsulation technology can be used in cosmetics to solve the solubility problem, that is, baicalin is encapsulated in the bilayer of liposomes using lecithin, polyols, etc., and the liposomes are soluble in water, but the baicalin improved by this technology still has difficulty in achieving the ideal bioavailability. The existing liposome encapsulation method has the problem of low drug loading, and cannot completely solve the problem that the raw material cannot be used due to the low amount added in skin care products. At the same time, the transdermal release ability of baicalin encapsulated by liposomes is poor, which affects the performance of various effects.

[0004] Therefore, there is an urgent need in the art to develop a baicalin modification technology with high transdermal efficiency, high bioavailability, and high drug loading. Summary of the invention

[0005] The purpose of the present invention is to provide baicalin microcapsules with high transdermal efficiency, high bioavailability and high drug loading, as well as a preparation method and application thereof.

[0006] In a first aspect of the present invention, a baicalin-cyclodextrin-vesicle complex is provided, comprising

[0007] (i) an active ingredient, the active ingredient comprising baicalin;

[0008] (ii) cyclodextrin; and

[0009] (iii) vesicle material, which comprises vesicles of microbial origin;

[0010] and,

[0011] The (ii) cyclodextrin includes the (i) active ingredient to form nanoparticles;

[0012] The (iii) vesicle material encapsulates the nanoparticles to form a baicalin-cyclodextrin-vesicle complex.

[0013] In another preferred embodiment, the microorganisms include strains of Saccharomyces and strains of Bacillus.

[0014] In another preferred embodiment, the strain of Saccharomyces is selected from the following group: Saccharomyces cerevisiae, ... or a combination thereof.

[0015] In another preferred embodiment, the Bacillus strain is selected from the following group: Bacillus subtilis.

[0016] In another preferred embodiment, the composite has the following components in parts by weight:

[0017] Active ingredient 12-18 parts

[0018] Vesicle material 70-90 parts

[0019] 3 to 7 parts of cyclodextrin.

[0020] In another preferred embodiment, the vesicle material has the following components in parts by weight:

[0021] Yeast fermentation products 50~60

[0022] Bacillus fermentation products 20-30.

[0023] In another preferred embodiment, the compound contains the following components:

[0024] Weight Preferred weight parts More preferably, Active ingredients 12～18 13～17 14～16 Vesicle material 70～90 75～85 78～83 Cyclodextrin 3～7 4～6 4～6。

[0025] In another preferred embodiment, the vesicle material contains the following components:

[0026] Weight Preferred weight parts More preferably, Yeast fermentation products 52～58 53～56 54～56 Bacillus fermentation products 22～27 23～26 23～25。

[0027] In another preferred embodiment, the baicalin-cyclodextrin-vesicle complex significantly increases the skin penetration depth of baicalin, and the significantly increased skin penetration depth of baicalin means that compared with the penetration depth P0 of baicalin in in vitro skin tissue, the penetration depth of the baicalin-cyclodextrin-vesicle complex in in vitro skin tissue is P1, and the ratio of the two (P1 / P0) is ≥3.

[0028] In another preferred embodiment, the penetration depth of the baicalin-cyclodextrin-vesicle complex in the in vitro skin tissue is P1, and compared with the penetration depth of baicalin in the in vitro skin tissue P0, the ratio of the two (P1 / P0) is ≥3, preferably ≥4, and more preferably ≥5.

[0029] In another preferred embodiment, the penetration depth of the baicalin-cyclodextrin-vesicle complex in the in vitro skin tissue is P1, and compared with the penetration depth P2 of the liposome-encapsulated baicalin in the in vitro skin tissue, the ratio (P1 / P2) is ≥1.5, preferably ≥2.

[0030] In another preferred embodiment, the penetration depth of the baicalin-cyclodextrin-vesicle complex in the dermis of the skin tissue in vitro is Z1, and the penetration depth of baicalin in the dermis of the skin tissue in vitro is Z0, and the ratio of the two (Z1 / Z0) is ≥5, preferably ≥7, and more preferably ≥10.

[0031] In another preferred embodiment, the penetration depth of the baicalin-cyclodextrin-vesicle complex in the dermis of the skin tissue in vitro is Z1, and the penetration depth of the liposome-encapsulated baicalin in the dermis of the skin tissue in vitro is Z2, and the ratio of the two (Z1 / Z2) is ≥1.5, preferably ≥2, and more preferably ≥2.5.

[0032] In another preferred embodiment, the baicalin-cyclodextrin-vesicle complex has an excellent transdermal release rate.

[0033] In another preferred embodiment, the excellent transdermal release rate refers to a cumulative release of ≥80% in 36 hours, preferably ≥85%; and / or a cumulative release of ≥45% in 8 hours, preferably ≥50%.

[0034] In another preferred embodiment, the baicalin-cyclodextrin-vesicle complex has no obvious burst release phenomenon within 0 to 8 hours.

[0035] In another preferred embodiment, the baicalin-cyclodextrin-vesicle complex has one or more of the following properties:

[0036] (f1) Has ROS scavenging ability;

[0037] (f2) reduce the expression of MMP-1;

[0038] (f3) increase the expression of TIMP-1;

[0039] (f4) having a sustained release effect;

[0040] (f5) Increased expression of CAT;

[0041] (f6) Increased expression of SOD;

[0042] (f7) Promote the growth of human fibroblasts;

[0043] (f8) Increase the expression of GSH and / or increase the ratio of GSH to GSSG.

[0044] In another preferred embodiment, the cyclodextrin is selected from α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, or a combination thereof.

[0045] In another preferred embodiment, the particle size of the baicalin-cyclodextrin-vesicle complex is 50 to 500 nm.

[0046] In another preferred embodiment, the polymer dispersion index of the baicalin-cyclodextrin-vesicle complex is 0.1-0.2.

[0047] In another preferred embodiment, the particle size of the baicalin-cyclodextrin-vesicle complex is 100-400 nm, preferably 100-300 nm.

[0048] In another preferred embodiment, the polymer dispersibility index of the baicalin-cyclodextrin-vesicle complex is 0.1 to 0.15.

[0049] In another preferred embodiment, the Zeta potential of the baicalin-cyclodextrin-vesicle complex is -10 to 0 mV, preferably -5 to 0 mV, and more preferably -5 to -2 mV.

[0050] In another preferred embodiment, the polymer dispersibility index of the baicalin-cyclodextrin-vesicle complex is 0.05-0.3.

[0051] In another preferred embodiment, the average particle size of the vesicle material is 20 to 500 nm, preferably 50 to 400 nm, more preferably 80 to 300 nm, such as 100 nm.

[0052] The second aspect of the present invention provides a method for preparing the baicalin-cyclodextrin-vesicle complex as described in the first aspect of the present invention, comprising the following steps:

[0053] (s1) providing (i) an active ingredient, wherein the active ingredient comprises baicalin; (ii) cyclodextrin; and (iii) a vesicle material, wherein the vesicle material comprises a vesicle derived from a microorganism;

[0054] (s2) mixing the active ingredient, cyclodextrin and vesicle material in step (s1) to obtain a baicalin-cyclodextrin-vesicle complex.

[0055] In another preferred embodiment, the vesicle material is prepared by the following steps:

[0056] (s1) in a first culture medium, after culturing Bacillus, adding yeast culture, centrifuging, and obtaining a precipitate;

[0057] (s2) crushing the precipitate obtained in step (s1) to obtain a vesicle material.

[0058] In another preferred example, the first culture medium is a liquid culture medium.

[0059] In another preferred embodiment, the first culture medium is selected from: nutrient broth medium.

[0060] In another preferred embodiment, the crushing is performed by homogenizing and crushing with a homogenizer.

[0061] In another preferred embodiment, in step (s2), the vesicle material needs to be separated and purified, and the separation and purification uses a solid chromatography column for separation and purification.

[0062] The third aspect of the present invention provides a use of the baicalin-cyclodextrin-vesicle complex as described in the first aspect of the present invention for preparing a preparation or a composition.

[0063] In another preferred embodiment, the preparation or composition is used for anti-aging and anti-oxidation.

[0064] In another preferred embodiment, the baicalin-cyclodextrin-vesicle complex is also used to prepare cosmetics.

[0065] In a fourth aspect, the present invention provides a cosmetic comprising

[0066] (c1) the baicalin-cyclodextrin-vesicle complex as described in the first aspect of the present invention; and

[0067] (c2) cosmetically acceptable carriers or excipients.

[0068] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0069] Figure 1 The particle size and distribution of baicalin microcapsules are shown.

[0070] Figure 2 The Zeta potential test results of baicalin microcapsules are shown.

[0071] Figure 3 Shown are the XRD comparison patterns of baicalin microcapsules.

[0072] Figure 4 A comparison of the skin penetration ability of baicalin microcapsules and liposomes is shown, a: epidermis; b: dermis; c: fluorescence penetration depth.

[0073] Figure 5 The results of the penetration depth of baicalin microcapsules and baicalin liposomes into the skin are shown.

[0074] Figure 6 The transdermal release rate of Scutellaria baicalensis microcapsules, baicalin and Scutellaria baicalensis liposomes is shown.

[0075] Figure 7 The ROS scavenging ability of baicalin, baicalin liposomes and baicalin microcapsules was shown.

[0076] Figure 8 The changes in GSH / GSSG of baicalin, baicalin liposomes, baicalin microcapsules, and materials are shown.

[0077] Fig. 9 Shown is the MMP-1 content of human skin fibroblast supernatant.

[0078] Fig.10 Shown are the TIMP-1 levels of human skin fibroblast supernatants.

[0079] Fig.11 The effects of baicalin, baicalin liposomes, and baicalin microcapsules on CAT activity are shown.

[0080] Fig.12 The effects of baicalin, baicalin liposomes, and baicalin microcapsules on SOD activity are shown.

[0081] Fig.13 Human fibroblast staining images of baicalin, baicalin liposomes, and baicalin microcapsules are shown. DETAILED DESCRIPTION

[0082] After extensive and in-depth research, a large number of experiments and screening, the inventors unexpectedly discovered a baicalin-cyclodextrin-vesicle complex for the first time, which has the characteristics of high transdermal efficiency, high bioavailability, and high drug loading, and can be applied to cosmetics and pharmaceutical products. On this basis, the present invention was completed.

[0083] the term

[0084] In order to more easily understand the present invention, some technical and scientific terms are specifically defined below. Unless otherwise clearly defined in this article, all other technical and scientific terms used herein have the meanings commonly understood by those of ordinary skill in the art to which the present invention belongs. Before describing the present invention, it should be understood that the present invention is not limited to the specific methods and experimental conditions, because such methods and conditions can be changed. It should also be understood that the terms used herein are intended only to describe specific embodiments, and are not intended to be restrictive, and the scope of the present invention will be limited only by the appended claims.

[0085] As used herein, the term “comprise” or variations thereof such as “include” or “comprising”, etc., are understood to include the stated elements or components but do not exclude other elements or components.

[0086] As used herein, the terms "material", "vesicle", "microcapsule material" and "vesicle material" are used interchangeably and all refer to vesicle materials of microbial origin, which in the present invention are the outer shell part of the baicalin-cyclodextrin-vesicle complex, encapsulating the cyclodextrin nanoparticles encapsulating baicalin.

[0087] Baicalin-cyclodextrin-vesicle complex

[0088] As used herein, the terms "baicalin microcapsules", "baicalin modified by natural vesicle technology", "baicalin-cyclodextrin-vesicle complex", and "complex of the present invention" are used interchangeably and all refer to baicalin prepared by the method of the present invention with high transdermal efficiency, high bioavailability, and high drug loading.

[0089] The baicalin-cyclodextrin-vesicle complex is formed by encapsulating baicalin with a biomembrane vesicle structure formed by yeast and bacillus fermentation products. The biomembrane vesicle structure can not only solve the problems of baicalin solubility, stability and other raw material applications, but also because it has a very similar structure to the human cell membrane, it is more helpful to improve the transdermal and intracellular ability of baicalin compared with the traditional liposome encapsulation method, so that baicalin can play a better antioxidant and anti-aging role in the skin care system.

[0090] The baicalin-cyclodextrin-vesicle complex of the present invention is a complex with uniform particle size and good safety, and its structure is: cyclodextrin nanoparticles containing baicalin are wrapped by vesicle materials, the vesicle materials are composed of yeast fermentation products and bacillus fermentation products, and the cyclodextrin nanoparticles containing baicalin are baicalin contained in the molecular cavity gap of cyclodextrin.

[0091] In the present invention, the baicalin-cyclodextrin-vesicle complex comprises

[0092] (i) an active ingredient, wherein the active ingredient is baicalin;

[0093] (ii) a vesicle material comprising a yeast fermentation product and a Bacillus fermentation product; and

[0094] (iii) cyclodextrin;

[0095] and,

[0096] The (ii) cyclodextrin includes the (i) active ingredient to form nanoparticles;

[0097] The (iii) vesicle material encapsulates the nanoparticles to form a baicalin-cyclodextrin-vesicle complex.

[0098] In another preferred embodiment, the mass ratio of active ingredient: vesicle material: cyclodextrin is 12-18:70-90:3-7.

[0099] In another preferred embodiment, the cyclodextrin is selected from α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, or a combination thereof.

[0100] The particle size of the baicalin-cyclodextrin-vesicle complex of the present invention is uniform, which is 50 to 500 nm, and the polymer dispersibility index is 0.05 to 0.3. The zeta potential of the baicalin-cyclodextrin-vesicle complex of the present invention is -10 to 0 mV, the potential distribution is uniform, the deviation range is small, there is no obvious dispersion of particles and material fragments, and it has good safety.

[0101] The microstructure of the baicalin-cyclodextrin-vesicle complex of the present invention is that baicalin is included in the cyclodextrin, and the vesicle material wraps the cyclodextrin including the baicalin.

[0102] The baicalin-cyclodextrin-vesicle complex of the present invention has an excellent skin penetration effect. The penetration depth of the baicalin-cyclodextrin-vesicle complex of the present invention in the in vitro skin tissue is P1. Compared with the penetration depth P0 of baicalin in the in vitro skin tissue, the ratio of the two (P1 / P0) is ≥3, preferably ≥4, and more preferably ≥5; compared with the penetration depth P2 of liposome-encapsulated baicalin in the in vitro skin tissue, the ratio of the two (P1 / P2) is ≥1.5, preferably ≥2.

[0103] The baicalin-cyclodextrin-vesicle complex of the present invention has an excellent transdermal release rate. In another preferred embodiment, the excellent transdermal release rate refers to no obvious burst release within 0 to 8 hours. In another preferred embodiment, the cumulative release within 36 hours is ≥80%, preferably 85%; and / or the cumulative release within 8 hours is ≥45%, preferably 50%.

[0104] Vesicle material

[0105] The vesicle material of the present invention includes vesicle material derived from microorganisms, including strains of the genus Saccharomyces and strains of the genus Bacillus.

[0106] In another preferred embodiment, the strain of Saccharomyces is selected from the group consisting of Saccharomyces cerevisiae, Saccharomyces cerevisiae, Saccharomyces cerevisiae, or a combination thereof. In another preferred embodiment, the strain of Bacillus is selected from the group consisting of Bacillus subtilis.

[0107] The vesicle material of the present invention can be obtained commercially or prepared by conventional methods. Typically, the vesicle material can be prepared by fermentation. A specific preparation method of a representative vesicle material is shown in patent WO2022055250.

[0108] MMP-1

[0109] Human skin fibroblasts are the main effector cells of dermal tissue, which can produce a variety of extracellular matrices such as collagen, laminin, and elastin. The water retained by fibroblasts and the ECM matrix they produce is the material basis for maintaining cell elasticity. Type I collagen is the most abundant protein in ECM, accounting for 90% of the total collagen content. It is the main component of the dermis and provides structural mechanical protection for surrounding tissues. MMP-1 is mainly involved in the degradation of type I collagen and type III collagen. When MMP-1 is overexpressed, it will specifically degrade ECM components and destroy the normal structure of collagen fibers and elastic fibers. Therefore, MMP-1 is one of the main enzymes that cause aging symptoms such as wrinkles and fine lines on the skin.

[0110] TIMP-1

[0111] Matrix metalloproteinase inhibitor-1 (TIMP-1) is one of the important glycoproteins in the skin and is involved in the degradation of the extracellular matrix. MMPs in the skin specifically degrade collagen. The excessive expression of MMPs promotes skin aging and destroys the skin ECM structure. In addition to inhibiting most known MMPs, TIMP-1 can also promote the proliferation of various cells and has an anti-apoptotic effect. Therefore, TIMP-1 is closely related to skin anti-aging.

[0112] GGH

[0113] Glutathione is a tripeptide that is indispensable for human metabolism. Its content is highest in infants and decreases with age. Glutathione has the effects of whitening and brightening, anti-oxidation, and increasing skin elasticity. It can inhibit the activity of tyrosinase, inhibit the production of melanin, reduce the division and biochemical activity of melanocytes, and enhance the role of vitamin C in the body; it can remove various free radicals in the human body, such as DPPH, hydroxyl free radicals, superoxide anions, etc.; it can integrate the skin's moisturizing environment and provide a good production environment for new skin cells, thereby making the skin smoother and more elastic; Glutathione is divided into oxidative GSSG and reductive GSH, of which GSH is the main active.

[0114] CAT

[0115] Catalase (CAT) is one of the main antioxidant enzymes in the human body. Its main functions include decomposing active oxygen (hydrogen peroxide, superoxide anions, etc.) produced during the body's metabolism, reducing free radicals and lipid peroxides in the human body, and is closely related to the oxidative aging of the human body.

[0116] SOD

[0117] SOD (superoxide dismutase) is an enzyme that catalyzes the conversion of superoxide into oxygen and hydrogen peroxide through dismutation reaction. It is widely present in the human body and is an important antioxidant enzyme. SOD can specifically remove free radicals in the skin, repair oxygen-damaged cells, and delay skin aging.

[0118] HSF

[0119] Human fibroblasts (HSF) are one of the main components of skin tissue. The fiber density, fiber thickness and fiber orientation of fibroblasts can largely reflect the state and degree of aging of cells. UV light will make cell collagen fibers thinner and accelerate cell aging.

[0120] Baicalin

[0121] Baicalin is a flavonoid compound extracted and separated from the dried root of Scutellaria baicalensis Georgi. It is almost insoluble in common cosmetic solvents such as water and ethanol. Baicalin has significant biological activity, antibacterial, diuretic, anti-inflammatory, anti-allergic and antispasmodic effects, and has strong anti-cancer effects and other physiological effects. It has an important position in clinical medicine. Baicalin can also absorb ultraviolet rays, scavenge oxygen free radicals, and inhibit the production of melanin. Therefore, it can be used in both medicine and cosmetics. It is a cosmetic raw material with good application prospects.

[0122] Cosmetic composition

[0123] The present invention also provides a cosmetic composition containing the baicalin-cyclodextrin-vesicle complex of the present invention. The cosmetic composition of the present invention comprises (a) the baicalin-cyclodextrin-vesicle complex of the present invention; and (b) a cosmetically acceptable carrier or excipient.

[0124] The baicalin-cyclodextrin-vesicle complex of the present invention can be prepared into various cosmetic compositions, such as emulsions, liquids, ointments, creams, pastes, cakes, powders and the like, by conventional methods.

[0125] To the extent that the effects of the present invention are not hindered, other ingredients commonly used in cosmetics may be added to the cosmetics of the present invention, such as film formers, oil-soluble gelling agents, organically modified clay minerals, resins, moisturizers, preservatives, antibacterial agents, flavors, salts, antioxidants, pH adjusters, chelating agents, cooling agents, anti-inflammatory agents, skin beautifying ingredients (whitening agents, cell activators, rough skin improving agents, blood circulation promoters, skin astringents, anti-seborrheic agents, etc.), vitamins, amino acids, nucleic acids, hormones, inclusion compounds, etc.

[0126] The oil-soluble gelling agent is a gelling agent selected from metal soaps such as aluminum stearate, magnesium stearate, and zinc myristate; amino acid derivatives such as N-lauroyl-L-glutamic acid and α, γ-di-n-butylamine; cyclodextrin fatty acid esters such as cyclodextrin palmitate, cyclodextrin stearate, and cyclodextrin 2-ethylhexanoate palmitate; sucrose fatty acid esters such as sucrose palmitate and sucrose stearate; benzylidene sorbitol derivatives such as monobenzylidene sorbitol and dibenzylidene sorbitol; organic modified clay minerals such as dimethylbenzyldodecyl ammonium montmorillonite clay and dimethyldioctadecyl ammonium montmorillonite clay, and the like. One or two or more kinds of gelling agents can be used as needed.

[0127] Moisturizers include: glycerin, sorbitol, propylene glycol, dipropylene glycol, 1,3-butylene glycol, glucose, xylitol, maltitol, polyethylene glycol, hyaluronic acid, chondroitin sulfate, pyrrolidone carboxylate, polyoxyethylene methyl glucoside, polyoxypropylene methyl glucoside, etc.

[0128] Antibacterial preservatives include: alkyl p-hydroxybenzoates, benzoic acid, sodium benzoate, sorbic acid, potassium sorbate, phenoxyethanol, etc. Antibacterial agents include: benzoic acid, salicylic acid, carbolic acid, sorbic acid, alkyl p-hydroxybenzoates, para-chlorometacresol, hexachlorophene, benzalkonium chloride, chlorhexidine chloride, trichloro-N-carbanilide, triclosan, photosensitizer, phenoxyethanol, etc.

[0129] Antioxidants include: tocopherol, butylated hydroxyanisole, butylated hydroxytoluene, phytic acid, etc., pH adjusters include: lactic acid, citric acid, glycolic acid, succinic acid, tartaric acid, dl-malic acid, potassium carbonate, sodium bicarbonate, ammonium bicarbonate, etc., chelating agents include alanine, sodium salt of ethylenediaminetetraacetic acid, sodium polyphosphate, sodium metaphosphate, phosphoric acid, etc., coolants include: L-menthol, camphor, etc., anti-inflammatory agents include: allantoin, glycyrrhetinic acid, glycyrrhizic acid, tranexamic acid, azulene, etc.

[0130] Skin beautification ingredients include: whitening agents such as placenta extract, arbutin, glutathione, and saxifrage extract; cell activators such as royal jelly, photosensitizer, cholesterol derivatives, and calf blood extract; skin roughness improvers; blood circulation promoters such as valerianamide, benzyl nicotinate, β-butoxyethyl nicotinate, capsaicin, gingerol, blister beetle tincture, ichthyol, caffeine, tannic acid, α-borneol, nicotinic acid tocopherol, inositol hexanicotinate, cycloamycin, cinnarizine, tolazoline, acetylcholine, verapamil, cepharanthin, and γ-oryzanol; skin astringents such as zinc oxide and tannic acid; anti-seborrheic acid agents such as sulfur, etc. Vitamins include: vitamin A oil, rosin oil, rosin oil acetate, rosin oil palmitate, etc.; vitamins such as riboflavin, riboflavin butyrate, and flavin adenine nucleotide Vitamin B2; Vitamin B6 such as pyridoxine hydrochloride, pyridoxine dioctanoate, pyridoxine tripalmitate, vitamin B12 and its derivatives, vitamin B15 and its derivatives, vitamin C such as L-ascorbic acid, L-ascorbyl dipalmitate, sodium L-ascorbic acid-2-sulfate, dipotassium L-ascorbyl phosphate; Vitamin D such as ergocalciferol and cholecalciferol; Vitamin E such as α-tocopherol, β-tocopherol, γ-tocopherol, dl-α-tocopherol acetate, dl-α-tocopherol nicotinic acid, dl-α-tocopherol succinate; Vitamin H; Vitamin P; Nicotinic acid such as niacin, benzyl nicotinate, niacinamide, etc.; Pantothenic acid such as calcium pantothenate, D-panthenol, pantothenic ethyl ether, acetyl pantothenic ethyl ether, etc.; Biotin, etc.

[0131] Amino acids include: glycine, valine, leucine, isoleucine, serine, threonine, phenylalanine, arginine, lysine, aspartic acid, glutamic acid, cystine, cysteine, methionine, tryptophan, etc.; nucleic acids include deoxyribonucleic acid, etc.; hormones include estradiol, vinyl estradiol, etc.

[0132] Preferred examples of the cosmetics of the present invention include: skin care cosmetics, color cosmetics, and UV protection cosmetics, such as basic cosmetics such as lotions, creams, lotions, sunscreens, mask materials, facial cleansers, and essences; and color cosmetics such as foundations, whitening powders, and blushes.

[0133] The form of the product is not particularly limited, and may be liquid, emulsion, cream, solid, paste, gel, powder, multi-layered, mousse, spray, or the like.

[0134] The present invention also provides a skin care method, comprising the steps of: applying the baicalin-cyclodextrin-vesicle complex of the present invention, or a cosmetic or care product containing the baicalin-cyclodextrin-vesicle complex of the present invention to an individual in need thereof.

[0135] In another preferred embodiment, the effective concentration range of the baicalin-cyclodextrin-vesicle complex is 100 μg / ml to 500 mg / ml.

[0136] In another preferred embodiment, the method is an anti-aging, anti-oxidation, whitening, anti-inflammatory and the like method.

[0137] The main advantages of the present invention include:

[0138] (1) For the first time, a baicalin-cyclodextrin-vesicle complex with high transdermal efficiency, high bioavailability, and high drug loading was provided;

[0139] (2) The baicalin-cyclodextrin-vesicle complex of the present invention has anti-aging, anti-oxidation, whitening, anti-inflammatory and other functions and can be used in cosmetics;

[0140] (3) The baicalin-cyclodextrin-vesicle complex of the present invention has a sustained release effect, which reduces the toxicity to cells in a short period of time and reduces the probability of allergic reactions;

[0141] (4) The method for preparing baicalin-cyclodextrin-vesicle complex of the present invention is simple to operate, low in cost, high in efficiency, simple in process, and suitable for industrial production;

[0142] (5) The baicalin-cyclodextrin-vesicle complex of the present invention can penetrate into the dermis layer in the skin tissue.

[0143] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. The experimental methods in the following examples where specific conditions are not specified are usually performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or under conditions recommended by the manufacturer. Unless otherwise indicated, percentages and parts are weight percentages and weight parts.

[0144] Comparative Example

[0145] The baicalin liposomes were prepared by the following method. The content of the baicalin liposomes is shown in Table 1:

[0146] Table 1

[0147] Element content / % water 41.45％ Dipropylene glycol 35.5％ Baicalin 8.5％ Hydrogenated lecithin (pc75) 7.0％ Caprylic / capric glycerides 4.5％ Polysorbate-80 1.5％ PEG-40 Hydrogenated Castor Oil 1.5％ Disodium EDTA 0.05％

[0148] 1. Weigh dipropylene glycol and hydrogenated lecithin and stir at 75℃ and 600r / min for 10min until there is no liquid particles;

[0149] 2. Add glyceryl caprylate to the above solution, stir at 75°C and 600r / min for 5min to make it uniform; (control the mass ratio of glyceryl caprylate to baicalin to be between 1:2 and 2:1);

[0150] 3. Add baicalin to the above solution; stir at 75°C and 600 r / min for 10 min until the solution becomes a yellow uniform liquid;

[0151] 4. Add disodium EDTA to the above solution; stir at 75°C and 600 r / min for 5 minutes until it becomes a yellow uniform liquid;

[0152] 5. Use microfluidizer to homogenize the above solution at 2000psi for three times, control the discharge temperature of the first and second homogenization to 50℃, and the third homogenization temperature to 15℃; obtain a yellow slightly viscous liquid;

[0153] 6. Centrifuge at 5000r / min for 10min to remove insoluble solid impurities.

[0154] The encapsulation efficiency of the prepared liposomes is 82.83%. The test method is as follows: Take 100 mg of the sample, place it in a volumetric flask and dissolve it in deionized water to a constant volume of 10 mL. Take 400 uL of the solution in a 50K ultracentrifugal filter tube, centrifuge at 12000 r / min for 10 min, and the encapsulated baicalin is retained on the filter membrane, and the free baicalin is filtered into the lower centrifuge liquid. Take 100 uL of the lower centrifuge liquid, dilute it 100 times with anhydrous ethanol, and then determine the content, and compare it with the standard curve to obtain the free baicalin (mg) in the sample.

[0155] Example 1 Preparation of baicalin microcapsules

[0156] Weigh an appropriate amount of cyclodextrin, place it in a colloid mill for grinding, then weigh baicalin and add it to the colloid mill. After fully mixing and grinding, add a large amount of deionized water to fully dissolve the evenly mixed cyclodextrin scutellaria powder, and homogenize the solution with a high-pressure microfluidizer at a homogenization pressure of 30 MPa. After homogenization several times, collect the solution, place it in a freeze dryer at -80°C to remove the solvent, and obtain baicalin cyclodextrin inclusion nanoparticles.

[0157] Dissolve the freeze-dried Bacillus subtilis powder in nutrient broth liquid (NB) medium. Transfer the dissolved bacterial solution to a sterile test tube containing a certain amount of liquid culture medium and mix and culture. Inoculate the activated strain into a conical flask filled with NB culture medium, shake and culture under appropriate conditions for standby. Transfer the Bacillus cells to NB liquid culture medium containing a certain amount of yeast powder for culture. After the culture is completed, collect the fermentation liquid and centrifuge to obtain the bacterial precipitate, add phosphate buffer to the precipitate according to a certain volume ratio and resuspend it, and pass the buffer solution containing the bacterial cells through a high-pressure microfluidizer at a homogenization pressure of 30MPa for homogenization and crushing. Pass the solution containing the bacterial fragments through a solid chromatography column to separate and purify the collected vesicle fraction.

[0158] Weigh an appropriate amount of vesicle fraction and dilute it with phosphate buffer, then weigh baicalin cyclodextrin inclusion nanoparticles, mix the two and put them into a sonicator for sonication, then put the liquid into a high-pressure extruder with nitrogen as the pressure source, extrude it through a polycarbonate membrane, then collect the bottom precipitate by centrifugation, dry and grind it into powder to obtain baicalin microcapsules.

[0159] Example 2 Physical properties of baicalin microcapsules

[0160] 2.1 Particle size and distribution of baicalin microcapsules

[0161] like Figure 1 As shown, the particle sizes of the three batches of baicalin microcapsules are basically distributed between 100-300nm, with a PDI value of 0.141, good distribution, and no obvious particles and fragments are precipitated, proving that the system is relatively stable, the particle distribution is uniform, and the Zeta average particle size is 145nm. The average particle size of the vesicle material without baicalin loading is 50-300nm.

[0162] 2.2 Potential detection of baicalin microcapsules

[0163] like Figure 2 As shown, the Zeta potential test results showed that the average potential of the three batches of baicalin microcapsules in the aqueous solution system was -2.8mV, and the absolute value was much smaller than -30mV which was toxic to cells. This showed that the raw materials had good safety in use, and the potential distribution was uniform with a small deviation range, indicating that there were no obvious particles and material fragments dispersed in the system.

[0164] 2.3 XRD analysis of baicalin microcapsules

[0165] like Figure 3 As shown, the characteristic peaks of baicalin in baicalin microcapsules were basically covered by the materials, while in the group where baicalin was directly physically mixed with the encapsulating materials (physical mixing refers to mixing according to the component ratio of baicalin microcapsules without going through the process flow and then stirring evenly), most of the characteristic peaks of baicalin were not covered, proving that baicalin was encapsulated by the materials rather than dispersed in the encapsulating materials.

[0166] Example 3 Skin penetration ability of baicalin microcapsules

[0167] 3.1 Methods

[0168] The experiment used fresh abdominal skin of three-month-old miniature pigs. After depilation, the fat layer and part of the dermis were scraped off with a scraper, and the skin was cut into pieces of about 3.14cm 2 The size of the circle is fixed on the permeation device with the stratum corneum facing upward. FITC-HA was used for the experimental fluorescein to crosslink baicalin with fluorescein (crosslinking method: an appropriate amount of baicalin was dissolved in acetic acid-sodium acetate buffer, and tyramine (p-hydroxyphenylethylamine) and sodium cyanoboride were added in sequence, followed by water bath, light-proof stirring reaction; FITC was added to tyramidate baicalin, and borate buffer was used as the labeling environment overnight; denaturing vertical plate polyacrylamide gel electrophoresis was used to detect and separate FITC-labeled baicalin. On the basis of the above, free baicalin and labeled baicalin were further separated and purified using a glucose gel column, and Tris-HCl was used for elution. FITC-labeled baicalin was obtained, and the drugs were loaded by liposome and microcapsule encapsulation technology respectively. The microcapsule-encapsulated drug, liposome-encapsulated drug and non-encapsulated drug were diluted to a concentration of 2‰ (2 mg / mL, calculated based on the baicalin concentration) with pH = 7.4 PBS buffer, and 0.5 mL of the above dilutions were added to the permeation device respectively, and placed in a dark place for 1 hour. After the end, the skin was removed and repeatedly rinsed with clean water until the surface fluorescence showed no discoloration. The surface moisture was dried with absorbent paper, and after slightly drying in a dark place, the slices were sliced ​​and placed under a fluorescence microscope to observe the penetration of fluorescence in the skin.

[0169] 3.2 Results

[0170] like Figure 4 and Figure 5 As shown, liposome encapsulation and microencapsulation compared with the PBS group (ie, unencapsulated baicalin) both improved the skin's ability to absorb the drug. The wider the fluorescent band, the deeper the drug penetration.

[0171] By comparing the penetration ability of the three into the epidermis and dermis of the skin, and the drug uptake ability of cells in different skin layers, it can be observed that the drug in the PBS group mainly stays on the surface of the epidermis, while the liposome-encapsulated and microcapsule-encapsulated drugs have reached the connective fiber tissue of the dermis, and the penetration depth of the microcapsule group in the dermis is deeper than that of the liposome group, and the dermis cells take up more drugs.

[0172] According to statistics, the average penetration depths of microencapsulated, liposome-encapsulated and PBS groups in the skin were 1347.52μm, 555.35μm and 264.39μm, respectively. The penetration depths of microencapsulated and liposome-encapsulated groups were 5.1 times and 2.1 times higher than those of the PBS group, respectively. The average penetration depth of microencapsulated baicalin was more than twice that of liposome-encapsulated baicalin.

[0173] The thickness of the epidermis is a, a=156.72±13.59 μm. After subtracting the thickness of the epidermis, the penetration depths of the dermis of the microcapsule encapsulation, liposome encapsulation and PBS groups are approximately 1090.8, 398.63, and 107.67 μm, respectively, indicating that the baicalin microcapsules of the present invention have unexpectedly excellent dermis penetration performance.

[0174] The above results indicate that baicalin microcapsules have better skin penetration ability.

[0175] Example 4 Transdermal Release Rate of Baicalin Microcapsules

[0176] 4.1 Methods

[0177] The experimental conditions were 32°C, the medium used isotonic phosphate buffer with pH = 7.4, and a transdermal diffusion instrument was used. The experiment set up baicalin group, baicalin liposome group and baicalin microcapsule group respectively, and three parallels were set up in each group. The skin used was 1kg of the back skin of New Zealand white rabbits. After the New Zealand rabbits were anesthetized by isoflurane breathing, they were injected with 1mL of potassium chloride solution of appropriate concentration and killed. After dissection, the back skin of the New Zealand rabbits was taken, the hair and fat layer were removed, and the skin was repeatedly rinsed with clean water, divided into appropriate sizes, placed in physiological saline, and stored at 4°C for use. The drugs used in the diffusion cell were diluted with isotonic buffer, and the drugs used in the diffusion cell were diluted with isotonic buffer. Among them, the concentration of baicalin group was 150μg / mL; the concentration of baicalin microcapsules was 1mg / mL (equivalent to 150μg / mL of baicalin), the concentration of baicalin liposomes was 6mgmL (equivalent to 150μg / mL of baicalin), the addition amount was 2mL, the receiving pool used 8mL of isotonic phosphate buffer (potassium chloride, sodium chloride) with pH=7.4, the skin was placed between the two, clamped and placed in the transdermal instrument; at 0.5, 1, 2, 4, 8, 12, 24, 36, 48, 54, and 60h, 0.5mL of solution was drawn, and then 0.5mL of permeate was added. After taking points at each time, the solution was placed in a UV spectrophotometer to detect OD 280nm , compare with the baicalin standard curve, calculate the concentration, calculate the transmittance, and draw a chart.

[0178] 4.2 Results

[0179] Depend on Figure 6 As can be seen from Table 2, the cumulative release of baicalin microcapsules reached 85.24% at 36 hours, and the release rate of baicalin reached 89.24% at 24 hours, and there was no significant change in the subsequent release rate. At 8 hours, the cumulative release rate of scutellaria microcapsules was 53.85%, and the cumulative release rate of baicalin original drug was 75.39%.

[0180] Table 2

[0181]

[0182]

[0183] The cumulative release rate of baicalin has an obvious burst release phenomenon in 0-8h, which may cause greater toxicity to skin cells and thus induce allergic reactions.

[0184] Scutellaria baicalensis microcapsules have added a certain sustained-release effect on the basis of baicalin original drug, which reduces the toxicity of the drug to cells in a short period of time, reduces the probability of skin allergic reactions, ensures the concentration required for the drug to exert its efficacy within a certain period of time, and provides better help for anti-oxidation, whitening and anti-aging effects.

[0185] Example 5 The repair effect of baicalin microcapsules on cells damaged by active oxygen

[0186] 5.1 Methods

[0187] The cells used in the experiment were L1929 mouse embryonic fibroblasts (a major cell of the dermis). When the cell growth density was appropriate and the condition was good, UV irradiation was performed for an appropriate time to construct a cell reactive oxygen damage model. After the stimulation, the original culture medium was discarded, and the cells were rinsed 3 times with PBS. The culture medium containing baicalin, baicalin liposomes, and baicalin microcapsules was added respectively (the dosage of baicalin group was 50 μg / mL, the dosage of baicalin liposomes was 2 mg / mL (equivalent to 50 μg / mL of baicalin), the dosage of baicalin microcapsules was 333.3 μg / mL (equivalent to 50 μg / mL of baicalin), and the dosage of microcapsule wrapping material was 283.3 μg / mL). After the addition was completed, the cells were incubated for 2 h, 4 h, and 8 h respectively. After the incubation, the DCFH-DA fluorescent probe was taken and diluted with serum-free culture medium at an appropriate multiple (the DCFH-DA fluorescent probe can freely pass through the cell membrane and has no fluorescence itself. When it reacts with intracellular enzymes to generate DCFH, it can no longer pass through the cell membrane, while intracellular reactive oxygen can oxidize DCFH to DCF. DCF shows green fluorescence at an excitation wavelength of 488 nm, which can indicate the level of intracellular reactive oxygen). The original culture medium was discarded, each dish was rinsed with PBS for 3 times, 1 mL of the diluted DCFH-DA solution was added to each dish to load the probe in situ, and the dish was placed in a cell culture incubator for incubation for 40 min. After the incubation was completed, the original culture medium was discarded, the dish was washed with PBS for 3 times, 1 mL of PBS buffer was added, and the dish was placed under a fluorescence microscope to take bright and dark field cell state images.

[0188] 5.2 Results

[0189] The repair effect of baicalin on ROS and cells damaged by ROS is shown in Figure 7 , where the fluorescence intensity is positively correlated with the intracellular ROS reactive oxygen level. The higher the fluorescence intensity, the higher the reactive oxygen level; the lower the green fluorescence, the lower the ROS level.

[0190] The ROS levels of the baicalin, baicalin liposome and baicalin microcapsule treatment groups all decreased with the extension of treatment time, and the fluorescence intensity of the three treatments at 8 hours was significantly lower than that at 2 hours. At 4 hours, the results showed that the degree of reduction of ROS levels by baicalin microcapsule was significantly better than that of the other two groups.

[0191] When the treatment time is the same, the ability to remove intracellular reactive oxygen species and the ability to repair cells after reactive oxygen damage are: baicalin microcapsules ≥ baicalin liposomes ≥ baicalin. At 8 hours, the intracellular reactive oxygen level in the baicalin microcapsule group was basically completely removed, while there was still a certain level in the baicalin pure drug group.

[0192] The inventors speculated that the reason for the above results is that both baicalin microcapsules and baicalin liposomes promoted the cellular uptake of baicalin, and thus the intracellular reactive oxygen scavenging ability was greatly improved within the same period of time. The cell uptake ability of baicalin microcapsules was stronger than that of baicalin liposomes, and the intracellular ROS scavenging ability of baicalin microcapsules was also higher than that of baicalin liposomes.

[0193] The above results indicate that baicalin microcapsules improve the uptake of baicalin by cells, can remove reactive oxygen species in cells, and have excellent antioxidant capacity.

[0194] Example 6 Effect of baicalin microcapsules on the changes of GSH / GSSG in the intracellular oxygen damage repair model

[0195] 6.1 Methods

[0196] The effect of baicalin on GSH / GSSG in oxygen-damaged cells (such as the oxygen damage model described in Example 5) was tested. Five groups were set up in the experiment (blank control, baicalin, baicalin microcapsules, baicalin liposomes, and materials), and the relative changes of GSH / GSSG in the drug-treated group and the blank group were compared.

[0197] Among them, the dosage of the baicalin group was 50 μg / mL, the dosage of baicalin liposomes was 2 mg / mL (equivalent to 50 μg / mL of baicalin), the dosage of baicalin microcapsules was 333.3 μg / mL (equivalent to 50 μg / mL of baicalin), and the dosage of microcapsule wrapping material was 283.3 μg / mL. The cells constructed in the previous step (the cell active oxygen damage model constructed in Example 5) were administered, and the GSH and GSSG detection kits were used to prepare the standard curves of total glutathione and GSSG, respectively, and the OD was detected by a microplate reader. 405nm , after testing the total protein content respectively, the contents of GSH and GSSG per unit protein were calculated.

[0198] 6.2 Results

[0199] Figure 8is the relative content change value of GSH / GSSG in the oxygen damage repair model after baicalin administration. Taking the blank group as the benchmark, it can be observed that the GSH / GSSG ratio in the material, baicalin, baicalin liposome and baicalin microcapsule groups increased by 3.36%, 35.79%, 20.98% and 50.96% respectively compared with the blank group.

[0200] Depend on Figure 8 Compared with the oxygen damage model (blank group), after the administration of baicalin-related drugs, GSH / GSSG increased to varying degrees. Taking GSH as a variable, it can be seen that baicalin-related drugs have excellent scavenging effects on intracellular free radicals and oxides, and have excellent repair effects on cell oxygen damage.

[0201] In terms of oxygen damage repair, baicalin microcapsules have certain advantages over baicalin liposomes and baicalin, which can also be seen in subsequent experiments. In fact, it can be observed that after oxygen-damaged cells were incubated with baicalin, baicalin liposomes, and baicalin microcapsules, the growth state of cells in the microcapsule group was better than that in the other groups.

[0202] The results of the blank baicalin microcapsule material on the repair of oxygen-damaged cells and the promotion of GSH were not obvious, which ruled out the material effect of the baicalin microcapsule group. The baicalin encapsulated in the microcapsule should have corresponding improvements in stability and cellular uptake.

[0203] Example 7 Effect of baicalin microcapsules on MMP-1 secretion in human skin fibroblasts

[0204] 7.1 Methods

[0205] In this experiment, blank, baicalin, baicalin microcapsules, material (microcapsules without baicalin) and baicalin liposome groups were set up to study the inhibitory effect of baicalin on cellular MMP-1 secretion.

[0206] Experimental procedure: According to the dosage of baicalin group of 20μg / mL; baicalin liposome dosage of 800μg / mL (equivalent to baicalin 20μg / mL), baicalin microcapsule dosage of 133.3μg / mL (equivalent to baicalin 20μg / mL, baicalin microcapsule material dosage of 113.3μg / mL) was applied to human fibroblasts, the drug was diluted with culture medium, the cell supernatant was discarded, PBS was washed 3 times, the corresponding drug was added, and the culture was continued for 24h. After the culture was completed, the cell supernatant was taken, centrifuged, and the supernatant was taken for MMP-1 test.

[0207] 7.2 Results

[0208] Depend on Fig. 9It can be seen that the MMP-1 content in the baicalin group, material group and baicalin microcapsule group decreased differently compared with the blank group, decreasing by 11.06%, 4.1% and 17.14% respectively, indicating that the four have a certain inhibitory effect on the secretion of MMP-1.

[0209] In summary, baicalin has a significant inhibitory effect on the secretion of MMP-1, and its inhibitory ability is further enhanced after encapsulation.

[0210] Example 8 Effect of baicalin microcapsules on TIMP-1 enzyme secretion in human skin fibroblasts

[0211] 8.1 Methods

[0212] In this experiment, baicalin, baicalin microcapsules, baicalin liposomes, blank and material groups were set up to study the effects of baicalin and related encapsulations on the content of TIMP-1 in the matrix of human skin fibroblasts.

[0213] Experimental method: Baicalin liposomes were administered at a dosage of 800 μg / mL (equivalent to 20 μg / mL of baicalin) and 133.3 μg / mL of baicalin microcapsules (equivalent to 20 μg / mL of baicalin, 113.3 μg / mL of baicalin microcapsule material) to human fibroblasts, the drugs were diluted with culture medium, the cell supernatant was discarded, PBS was washed three times, the corresponding drugs were added, and the culture was continued for 24 hours. After the culture was completed, the cell supernatant was taken, centrifuged, and the supernatant was taken for MMP-1 test.

[0214] 8.2 Results

[0215] like Fig.10 As shown, the TIMP-1 content in the cell supernatant of the baicalin group, material group and baicalin microcapsule group increased to varying degrees compared with the blank group, increasing by 8.16%, 17.21% and 18.04%, respectively.

[0216] The TIMP-1 content in the baicalin liposome group decreased to a certain extent, about 12.42%, compared with the blank group. Some substance in the baicalin liposome material may affect the expression of TIMP-1.

[0217] The microcapsule material significantly promoted the expression of TIMP-1 in the extracellular matrix of human skin fibroblasts. The promoting effect of baicalin microcapsules on TIMP-1 expression was significantly stronger than that of unencapsulated baicalin, which increased by 2.21 times.

[0218] Combined with the data of MMP-1, baicalin can inhibit the expression of MMP-1 and increase the expression of TIMP-1, and has excellent anti-aging effects. The anti-aging effect of baicalin encapsulated in microcapsules is even better.

[0219] Example 9 Effect of baicalin microcapsules on the activity of intracellular CAT

[0220] 9.1 Methods

[0221] After baicalin, baicalin microcapsules, baicalin liposomes and materials were incubated with cells, the relative activity changes of CAT in the cells were tested. The experimental settings included five groups (blank control, baicalin, baicalin microcapsules, baicalin liposomes and materials).

[0222] The dosage of baicalin group was 50 μg / mL, the dosage of baicalin liposome was 2 mg / mL (equivalent to 50 μg / mL of baicalin), the dosage of baicalin microcapsule was 333.3 μg / mL (equivalent to 50 μg / mL of baicalin), and the dosage of microcapsule wrapping material was 283.3 μg / mL. After the administration, the CAT activity per unit protein content was detected and calculated, and the blank group was used as a benchmark to draw the CAT activity graph of the administration group.

[0223] 9.2 Results

[0224] like Fig.11 As shown, compared with the blank group, the CAT activity per unit protein content of the material, baicalin, baicalin liposome and baicalin microcapsule groups increased by 6.84%, 74.72%, 114.20% and 166.99%, respectively.

[0225] It can be observed that baicalin can significantly increase the activity of CAT, so baicalin has excellent antioxidant effect.

[0226] The effects of baicalin encapsulated by liposomes and microcapsules on CAT activity were significantly better than those of baicalin not encapsulated, and microcapsule encapsulation was better than liposome encapsulation.

[0227] Combined with the in vitro release experiment, the stability and biological activity of baicalin after encapsulation have been improved to a certain extent. In addition, the sustained-release effect after encapsulation enables baicalin to be released stably and for a long time, which is also one of the reasons for the better effect after encapsulation.

[0228] Example 10 Effect of baicalin microcapsules on the activity of intracellular SOD

[0229] 10.1 Methods

[0230] After incubating baicalin, baicalin microcapsules, baicalin liposomes and materials with cells, the relative activity of intracellular SOD changed. The experimental settings included five groups (blank control, baicalin, baicalin microcapsules, baicalin liposomes and materials).

[0231] The dosage of baicalin group was 50 μg / mL, the dosage of baicalin liposome was 2 mg / mL (equivalent to 50 μg / mL of baicalin), the dosage of baicalin microcapsule was 333.3 μg / mL (equivalent to 50 μg / mL of baicalin), and the dosage of microcapsule wrapping material was 283.3 μg / mL. After the administration, SOD activity test was performed, and the SOD activity graph of the administration group was drawn with the blank group as the benchmark.

[0232] 10.1 Results

[0233] like Fig.12 As shown, the SOD activity per unit protein content of the material, baicalin, baicalin liposome and baicalin microcapsule groups were increased by 0.89%, 104.31%, 23.92% and 197.45% respectively compared with the blank control group. The SOD activity per unit protein was significantly improved after the administration of baicalin.

[0234] The rate of SOD activity improvement after baicalin microcapsule encapsulation was significantly improved compared with that of baicalin without encapsulation, but the promoting effect of baicalin encapsulated by liposomes on SOD activity was somewhat lower than that of baicalin without encapsulation.

[0235] The blank microcapsule material group did not show any promoting effect on SOD activity, which can rule out the influence of the material in the baicalin microcapsules on the experiment. Therefore, microencapsulation greatly improved the expression and activity-promoting effect of baicalin on intracellular SOD.

[0236] Combined with the fact that the activities or contents of SOD, CAT and GSH increased to varying degrees after administration, it can be seen that baicalin has a high-capacity and comprehensive effect on the antioxidant effect of microcapsule cells.

[0237] Example 11 Baicalin microcapsules repair light damage of human fibroblasts (HSF) and promote fiber growth

[0238] 11.1 Methods

[0239] The experiment used human fibroblasts (HSF), and set up blank control, baicalin, baicalin microcapsules, and baicalin liposome groups. Among them, the dosage of baicalin group was 20μg / mL; the dosage of baicalin liposome was 800μg / mL (equivalent to baicalin 20μg / mL), and the dosage of baicalin microcapsules was 133.3μg / mL (equivalent to baicalin 20μg / mL). The cells were irradiated with ultraviolet rays of appropriate intensity to establish a light damage model, and then the above drugs were diluted with serum-free culture medium, added to the culture dish, and cultured for 48 hours at 37℃ and 5% CO2. After the end, they were fixed with polyformaldehyde, stained with hematoxylin, ponceau-fuchsin, and aniline blue dye, respectively, and the fiber density, direction, and thickness were observed under a microscope, and the unit cell fiber area was calculated and a table was drawn.

[0240] 11.2 Results

[0241] like Fig.13 As shown, blue represents human fibroblast cell fibers and red represents fibrocytes.

[0242] After ultraviolet irradiation, the cell fibers in the blank control group became thinner and abnormal, indicating that ultraviolet irradiation accelerated cell aging.

[0243] After incubating UV-damaged cells with baicalin, baicalin liposomes and baicalin microcapsules, it can be clearly observed that the cell viability was significantly improved, the direction of cell fibers returned to normal, and the cell fibers became significantly thicker than before incubation.

[0244] The results showed that baicalin, baicalin liposomes and baicalin microcapsules all had a repairing effect on photoaged fibroblasts, proving the anti-aging effect of baicalin on the dermis.

[0245] The above results show that the particle size and particle distribution of baicalin microcapsules are uniform, there are no obvious particles and material fragments, the system is stable, and it has excellent transdermal release ability, skin penetration ability, antioxidant ability, and anti-aging ability, and can be used for anti-aging and anti-oxidation of the dermis.

[0246] discuss

[0247] In cosmetic applications, the occurrence of oxidative aging-related problems is mainly concentrated in the dermis of the skin, so whether the raw materials can be delivered to the dermis and the amount of intake by the cells measured in the dermis are crucial to the antioxidant and anti-aging effects.

[0248] The present invention provides baicalin microcapsules. In terms of transdermal penetration depth and cumulative release rate, the data of baicalin microcapsules are superior to those of baicalin original drug and baicalin liposomes. Both liposome-encapsulated and microcapsule-encapsulated drugs have reached the connective fiber tissue of the dermis, but the penetration depth of the microcapsule group in the dermis is deeper than that of the liposome group, and the dermis cells take up more drugs.

[0249] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A baicalin-cyclodextrin-vesicle complex, characterized in that: Include (i) an active ingredient, the active ingredient comprising baicalin; (ii) cyclodextrin; and (iii) vesicle material, said vesicle material comprising vesicles of microbial origin; and, The (ii) cyclodextrin includes the (i) active ingredient to form nanoparticles; The (iii) vesicle material wraps the nanoparticles to form a baicalin-cyclodextrin-vesicle complex.

2. The baicalin-cyclodextrin-vesicle complex according to claim 1, wherein The microorganisms include strains of the genus Saccharomyces and strains of the genus Bacillus.

3. The baicalin-cyclodextrin-vesicle complex according to claim 1, wherein The composite has the following components in parts by weight: 12-18 parts active ingredient 70-90 parts of vesicle material 3 to 7 parts of cyclodextrin.

4. The baicalin-cyclodextrin-vesicle complex according to claim 1, wherein The baicalin-cyclodextrin-vesicle complex significantly increases the skin penetration depth of baicalin. The significantly increased skin penetration depth of baicalin means that compared with the penetration depth P0 of baicalin in in vitro skin tissue, the penetration depth of the baicalin-cyclodextrin-vesicle complex in in vitro skin tissue is P1, and the ratio of the two (P1 / P0) is ≥3.

5. The baicalin-cyclodextrin-vesicle complex according to claim 1, wherein The cyclodextrin is selected from α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, or a combination thereof.

6. The baicalin-cyclodextrin-vesicle complex according to claim 1, wherein The particle size of the baicalin-cyclodextrin-vesicle complex is 50-500 nm.

7. The baicalin-cyclodextrin-vesicle complex according to claim 1, wherein The polymer dispersion index of the baicalin-cyclodextrin-vesicle complex is 0.1-0.

2.

8. A method for preparing the baicalin-cyclodextrin-vesicle complex according to any one of claims 1 to 7, characterized in that: The following steps are involved: (s1) providing (i) an active ingredient comprising baicalin; (ii) cyclodextrin; and (iii) a vesicle material comprising a vesicle of microbial origin; (s2) mixing the active ingredient, cyclodextrin and vesicle material in step (s1) to obtain a baicalin-cyclodextrin-vesicle complex.

9. Use of the baicalin-cyclodextrin-vesicle complex according to any one of claims 1 to 7, characterized in that: For preparing a preparation or composition.

10. A cosmetic, characterized in that: The cosmetic comprises (c1) the baicalin-cyclodextrin-vesicle complex according to any one of claims 1 to 7; and (c2) cosmetically acceptable carriers or excipients.

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