Metal hydroxide compound and preparation method thereof

Abstract

Provided is a metal hydroxide composite including a base layer, a surface layer, and an active ingredient, the metal hydroxide composite including a multilayer hydroxide structure of Chemical Formula 1; a metal hydroxide composite prepared by including a step of co-precipitating an active ingredient and a metal hydroxide structure precursor by a precipitation reaction using an alcohol and water, and a method for preparing the same.

Description

[0001] This application is a divisional application of an application with the application number “201980024676.5” (international application number “PCT / KR2019 / 012331”) filed on September 23, 2019, the title of which is “Metal Hydroxide Composite Including Modified Multilayered Hydroxide Structure Containing Active Ingredient and Preparation Method Thereof”. TECHNICAL FIELD

[0002] The present application relates to a metal hydroxide composite including a modified multilayered hydroxide structure including a base layer, a surface layer, and an active ingredient, and a preparation method thereof. BACKGROUND

[0003] Active ingredients of a cosmetic that can be applied into the skin and can directly exhibit its effects can include vitamins, etc. Vitamins are substances necessary in the body, which have abilities such as promoting metabolism of living organisms, exhibiting an antioxidant effect, protecting cell walls, enhancing immunity, increasing resistance to infection, etc., and in most primates, biosynthesis of vitamins cannot be performed, intake through food is essential, and insufficient intake of vitamins causes various vitamin deficiencies. In addition, in skin beauty and treatment, vitamins play a very important role in maintaining healthy skin, such as preventing pigmentation, promoting collagen synthesis, blocking ultraviolet (UV) radiation, preventing skin dryness and keratinization, preventing wrinkles, moisturizing the skin, etc. Examples of vitamins include retinol (vitamin A), ascorbic acid (vitamin C), tocopherol (vitamin E), and derivatives thereof.

[0004] In addition to vitamins, active ingredients of a cosmetic can include alpha-hydroxy acids (AHAs) such as lactic acid, citric acid, and salicylic acid, which have a function of promoting metabolism of the skin by removing the stratum corneum of the skin; kojic acid (5-hydroxy-2-(hydroxymethyl)-4H-pyran-4-one), which has a function of whitening the skin by inhibiting melanin biosynthesis; indole-3-acetic acid (C 10 H9NO2), which prevents wrinkles by promoting fibroblast proliferation activity; and salicylic acid (2-hydroxybenzoic acid, C7H6O3), which has an antioxidant and acne treatment function; and various other materials known as cosmetic ingredients.

[0005] However, most of the above-mentioned cosmetic ingredient materials are limited in practical application because of the material's own stability, skin irritation and toxicity, slow release, dispersing ability, etc., and thus their functions and effects are not fully exhibited. For example, vitamins are physically and chemically very unstable and are easily destroyed by heat, light, moisture, oxygen, alkali, etc., so their functions and effects are reduced, or they are discolored or produce an odor. In addition, α-hydroxy acids such as lactic acid have the problem that they cause skin irritation. In the case of kojic acid, melanin or melanocytes are degraded due to the property of penetrating the basal layer, and thus, when used at a high concentration, it causes various skin diseases such as dermatitis, skin cancer, etc., and can be discolored due to oxidation in the case of light and high temperature. Indole-3-acetic acid has poor resistance to external environments, heat, light, moisture, oxygen, etc., and is particularly sensitive to light, which leads to the problems of discoloration and odor.

[0006] Therefore, in order to obtain the effect of stabilizing active substances, reducing skin irritation or toxicity, etc., thorough research has been conducted. Specifically, Korean Patent No. 115076 discloses a method of preparing vitamin nanocapsules by impregnating and encapsulating inner phase microdroplets with vitamins and various active ingredients, and again stabilizing the active substances using a double lipid membrane and a water-dispersing polymer. In addition, Korean Patent Application Publication No. 2000-0048451 discloses a method of encapsulating the core of an oil-soluble active ingredient droplet with a synthetic and natural water-dispersing anionic polymer that is insoluble in water.

[0007] However, the foregoing methods are time-consuming due to the complex method for treating active substances, processes, etc., and thus are not economical. In addition, since coating of the active ingredient must be additionally performed, the effect of the individual active ingredient is reduced, and there is a problem that excess active material is not included in the complex.

[0008] (Patent Document 1) Korean Patent No. 115076

[0009] (Patent Document 2) Korean Patent Application Publication No. 2000-0048451. SUMMARY

[0010] TECHNICAL PROBLEM

[0011] The present invention has been made to solve the problems encountered in the prior art, and is intended to provide a metal hydroxide composite comprising a multilayer hydroxide structure including a base layer, a surface layer, and an active ingredient, wherein the metal hydroxide composite can stably include an excess of the active ingredient alone so that the metal hydroxide composite can maximize the effect of the active ingredient; and is intended to provide a method for preparing the metal hydroxide composite, which is economical by shortening the preparation process because the active ingredient can be included in a more stable and effective manner.

[0012] Technical Solution

[0013] The present invention provides a metal hydroxide composite, which includes a modified multilayer hydroxide structure of the following Chemical Formula 1. The modified multilayer hydroxide structure includes a base layer, a surface layer, and an active component.

[0014] [Chemical Formula 1]

[0015] {[M x (OH) z ] B [M y (OH) w ] S}{(A n ) q}·m(H2O)

[0016] In this paper, [M x (OH) z ] B represents the base layer of the multilayer hydroxide structure, and [M y (OH) w ] S represents the surface layer of the multilayer hydroxide structure,

[0017] M is selected from Ca 2+ Mg 2+ 、Cu 2+ 、Zn 2+ 、Ni 2+ 、Co 2+ and Fe2 + Any one of the divalent metal cations in the group consisting of,

[0018] x is 0.6 to 3,

[0019] y is 0 to 2,

[0020] z is 1 to 5,

[0021] w is 0 to 4,

[0022] z+w is 1 to 9,

[0023] q is 1 to 4,

[0024] m is 0.1 to 10,

[0025] n is the number of charges of A, and

[0026] A represents an active ingredient, wherein A is an anionic compound, an anionic compound as a functional group having an electrostatic attraction depending on pKa, the anionic compound including at least one selected from the group consisting of a hydroxyl group (-OH), a carbonyl group (-CO-), an aldehyde group (-CHO), a carboxyl group (-COOH), a sulfuric acid group (-SO3 2- ), a dihydrogen phosphate group (-H2PO4 2- ), and a phosphate group (-PO4 3- ).

[0027] The present application provides a metal hydroxide complex prepared by subjecting an active ingredient and a metal hydroxide to coprecipitation using an alcohol and water through a precipitation reaction.

[0028] The present application provides a method of preparing a metal hydroxide complex, including subjecting an active ingredient and a metal hydroxide to coprecipitation using an alcohol and water through a precipitation reaction.

[0029] Advantageous Effects

[0030] According to the present application, an active ingredient can be more abundantly included in a multilayer hydroxide structure, have excellent sustained-release properties, and be included in a metal hydroxide complex in a stable state and in a large amount, thereby improving the effect of the delivered active ingredient.

[0031] In addition, the metal hydroxide complex stably includes an unstable active ingredient, thereby enabling long-term storage of the active ingredient, and when it is delivered to the skin, enabling minimization of irritation, thereby providing an irritation-free effect.

[0032] In the present application, an active ingredient is included in a multilayer hydroxide structure, thereby showing an excellent collagen expression effect. BRIEF DESCRIPTION OF DRAWINGS

[0033] Figure 1 is an XRD pattern of each material of Table 1, wherein is an MZLA pattern, is a reference pattern, and is a zinc basic salt pattern;

[0034] Figure 2is an XRD pattern of metal hydroxide depending on the ratio of ethanol to water solvent, wherein, when the sum of the weight ratios of ethanol to water is 10, The graph for ethanol 1.0: water 9.0 is shown. The figure for ethanol 4.6: water 5.4, The graph for ethanol 6.4: water 3.6 and It is the graph at ethanol 8.4: water 1.5;

[0035] Figure 3 is the XRD pattern, where and For use with ethanol:water ( Ethanol: water = 85:15, and XRD patterns of metal hydroxides in a mixed solvent of ethanol (4.6): water (5.4), is the XRD pattern of the metal hydroxide using water alone as the solvent, and is the XRD pattern of ZBS-NO3;

[0036] Figure 4 shows XRD patterns of metal hydroxides according to the ratio of methanol:water solvent;

[0037] Figure 5 shows XRD patterns of metal hydroxides according to the ratio of ethanol:water solvent;

[0038] Figure 6 shows XRD patterns of metal hydroxides according to the ratio of propanol:water solvent;

[0039] Figure 7 shows XRD patterns of metal hydroxides according to the ratio of butanol:water solvent;

[0040] Figure 8 SEM images of metal hydroxides according to the ratio of ethanol to water solvent are shown, and the SEM images are enlarged images of a portion of the surface of a single particle;

[0041] Figure 9 shows SEM images of metal hydroxides according to the ratio of ethanol:water solvent;

[0042] Figure 10 shows SEM images of metal hydroxides according to the ratio of ethanol:water solvent;

[0043] Figure 11 Shows the specific configuration of the metal hydroxide complex according to the present invention, [M x (OH) z ] B Representing the grassroots, and [M y (OH)w ] S representative surface layer;

[0044] Figure 12 shows the measurement results of cell viability of human dermal fibroblasts treated with the metal hydroxide complex of the present application, the horizontal axis represents the concentration of the treated sample material, and the vertical axis represents the cell viability;

[0045] Figure 13 shows the change in the expression of collagen type 1 gene (COL1A1) of human dermal fibroblasts treated with the metal hydroxide complex of the present application, the horizontal axis represents the concentration of the treated sample material, and the vertical axis represents the expression level of COL1A1 mRNA;

[0046] Figure 14 shows the change in the expression of collagen type 3 gene (COL3A1) of human dermal fibroblasts treated with the metal hydroxide complex of the present application, the horizontal axis represents the concentration of the treated sample material, and the vertical axis represents the expression level of COL3A1 mRNA;

[0047] Figure 15 shows the solubility of Zn;

[0048] Figure 16 shows the solubility curve of zinc hydroxide;

[0049] Figure 17 shows the solubility of zinc hydroxide, *S is the solubility represented by the mole of zinc per kilogram of water; and

[0050] Figure 18 is an XRD pattern of the metal hydroxide according to the ratio of ethanol: water solvent. DETAILED DESCRIPTION

[0051] The present application relates to a metal hydroxide complex including a modified multilayer hydroxide structure of the following Chemical Formula 1, the modified multilayer hydroxide structure including a base layer, a surface layer, and an active ingredient.

[0052] [Chemical Formula 1]

[0053] {[M x (OH) z ] B x (OH) z ] B [M y (OH) w ] S}{(A n ) q}·m(H2O)

[0054] Herein, [M x (OH) z ] Brepresents a base layer of a multi-layered hydroxide structure, and [M y (OH) w ] S represents a surface layer of a multi-layered hydroxide structure,

[0055] M is any one of a divalent metal cation selected from the group consisting of Ca 2+ , Mg 2+ , Cu 2+ , Zn 2+ , Ni 2+ , Co 2+ , and Fe 2+ ,

[0056] x is 0.6 to 3,

[0057] y is 0 to 2,

[0058] z is 1 to 5,

[0059] w is 0 to 4,

[0060] z+w is 1 to 9,

[0061] q is 1 to 4,

[0062] m is 0.1 to 10,

[0063] n is the number of charges of A, and

[0064] A represents an active ingredient, wherein A is an anionic compound including at least one selected from the group consisting of a hydroxyl group (-OH), a carbonyl group (-CO-), an aldehyde group (-CHO), a carboxyl group (-COOH), a sulfuric acid group (-SO3 2- ), a dihydrogen phosphoric acid group (-H2PO4 2- ), and a phosphoric acid group (-PO4 3- ) as a functional group having electrostatic attraction depending on pKa.

[0065] The active ingredient represented by A includes at least one selected from the group consisting of a hydroxyl group (-OH), a carbonyl group (-CO-), an aldehyde group (-CHO), a carboxyl group (-COOH), a sulfuric acid group (-SO3 2- ), a dihydrogen phosphoric acid group (-H2PO4 2- ), and / or a phosphoric acid group (-PO4 3- ).

[0066] More specifically, Chemical Formula 1 can be represented by the following: {[Zn3(OH)4] B [Zn2(OH)3] S}{(ascorbic acid)2}·2(H2O), {[Zn3(OH)4] B[Zn1(OH)2] S}{(ascorbic acid)2}·3(H2O), {[Zn3(OH)4] B [Zn1(OH)1] S}{(ascorbic acid)2}·3(H2O), and {[Zn3(OH)3] B [Zn1(OH)1] S}{(ascorbic acid)2}·2(H2O).

[0067] A is an active ingredient, which can be an anion, and examples of the active ingredient having a functional group with electrostatic attraction depending on pKa, especially a hydroxyl group (-OH) can include ascorbic acid, 4-n-butylresorcinol, tocopherol, bakuchiol, (+)-catechin, curcumin, hydroxytyrosol, phytol, resorcinol, safflor yellow, luteolin, corilagin, piceid, retinol, rutin, hydroquinone, asiaticoside, madecassoside, ginsenoside, borneol, dihydroeugenol, aspalathin, eugenol, garcinol, pelargonidin, petunidin, cyaniding, delphinidin, peonidin, lutein, quercetin, adenosine, ascorbyl palmitate, ascorbyl glucoside, pyridoxine, thiamine, saponin, secoisolaricirecinol, martisianol, pinoresinol, pinocembranol, lariciresinol, syringaresinol, artigenin, ent-lactone, and ent-diol.

[0068] Examples of the active ingredient including a carboxyl group (-COOH) can include arachidonic acid, abietic acid, abscisic acid, alpha-lipoic acid, azelaic acid, caffeic acid, hydroxybenzoic acid, protocatechuic acid, ellagic acid, ferulic acid, fulvic acid, oleanolic acid, phenolic acid, hydroxycinnamic acid, vanillic acid, protocatechuic acid, salvianolic acid, sinapic acid, thromboxane, valeric acid, veratric acid, chlorogenic acid, asiatic acid, madecassic acid, suberic acid, hyaluronic acid, ursolic acid, ascorbic acid, salvianolic acid B, pyridine-3-carboxylic acid, ascorbyl palmitate, ascorbyl glucoside, carnitine, pantothenic acid, biotin, folic acid, allitridin, glutathione, serine, glycine, alanine, avenanthramide, threonine, cysteine, valine, leucine, methionine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, histidine, lysine, and arginine, etc.

[0069] Examples of the active ingredient including an aldehyde group (-CHO) can include decanal, retinal, cinnamaldehyde, catechol aldehyde, coniferyl aldehyde, eugenol, and vanillin, etc.

[0070] Examples of active ingredients including a carbonyl group (-CO-) can include edaravone, idebenone, coenzyme Q10, ubiquinone, MitoQ, astaxanthin, caffeine, paraxanthine, theophylline, matairesinol, physcion, propanone, coumarin, genistein, chalcones, naringenin, bergenin, amentoflavone, biochanin A, riboflavin, and sesamin, among others.

[0071] Including sulfate group (-SO3 2- Examples of the anionic active ingredient of ) may include dextran sulfate, ferrous sulfate, ferulic acid-4-O-sulfate, and ascorbic acid sulfate, etc.

[0072] And includes dihydrogen phosphate (-H2PO4 2- Examples of the compound of the present invention may include adenosine monophosphate, retinol phosphate, and ascorbyl palmitate phosphate.

[0073] And includes phosphate group (-PO4 3- Examples of the compound of ) may include adenosine diphosphate, adenosine triphosphate, and ascorbyl phosphate, etc.

[0074] More specifically, the active ingredient preferably includes at least one selected from the group consisting of ascorbic acid, cysteine, salicylic acid, abscisic acid, serine, alanine, glycine, carnitine, isoleucine, leucine, suberic acid, proline, valine, azelaic acid, phenylalanine, caffeic acid, tryptophan, coumarin, tyrosine, aspartic acid, ferulic acid, glutamic acid, arginine, pyridine-3-carboxylic acid, histidine, lysine, ascorbyl sulfate, threonine , protocatechuic acid, ascorbyl phosphate, methionine, valeric acid, asparagine, glutamine, vanillic acid, hydroxybenzoic acid, cinnamaldehyde, pantothenic acid, biotin, retinol, α-lipoic acid, resveratrol, 4-n-butylresorcinol, hydroxytyrosol, pyridoxine, sinapinic acid, coniferyl aldehyde, syringaldehyde and theophylline, so that the powder X-ray diffraction pattern can have peaks at diffraction angles (2θ) of 5.96±1°, 33.46±1° and 59.3±1°.

[0075] In the present invention, the multilayer structure means that the base layer and / or the surface layer can be modified in the structural form of the metal hydroxide generally formed, and more specifically, in the metal hydroxide structure, the structural form of the metal hydroxide of the surface layer between the base layer and the surface layer can be more variable. Specifically, the base layer and the surface layer are Figure 11The multi-layer structure can include a modified multi-layer structure configured such that a conventional metal hydroxide structure having a constant double-layer structure arrangement is gradually formed into an asymmetric layered structure, thereby inducing modification of the double-layer structure. This means that the surface layer structure is modified, is such that individual metal hydroxide structures are formed in different sizes and stacked to form a multi-layer structure. The multi-layer structure can have a 2 to 5 layer, preferably 3 layer structure.

[0076] In the present invention, the multi-layer hydroxide structure can have a powder X-ray diffraction pattern having peaks at diffraction angles (2q) of 5.96±1°, 33.46±1° and 59.3±1°. Herein, the multi-layer hydroxide structure can include an asymmetric and / or modified layered structure. More preferably, it can have peaks at (2q) of 5.96±0.5°, 33.46±0.5° and 59.3±0.5°, when the above peaks are satisfied, the metal hydroxide composite can be provided in the form of a symmetric and / or modified layered structure (2 to 5 layers).

[0077] In the present invention, the multi-layer hydroxide structure can have a powder X-ray diffraction pattern having peaks at diffraction angles (2q) of 5.96±1°, 8.5±1°, 10.36±1°, 13.36±1°, 19±1°, 20.84±1°, 21.7±1°, 26.34±1°, 37.68±1°, 31.48±1°, 33.78±1°, 34.88±1° and 59.3±1°. When the above peaks are satisfied, the metal hydroxide composite can be provided in the form of a layered structure, and the layered structure can include a modified asymmetric layered structure. More specifically, ethanol and / or water can be used as a solvent to form the layered structure. By adjusting the solvent ratio, an asymmetric layered structure and / or a modified layered structure can be formed, and the formed asymmetric layered structure and / or modified layered structure can be a layered structure having multiple layers, for example, 2 to 5 layers.

[0078] In the present invention, the multi-layer hydroxide structure can have a powder X-ray diffraction pattern having peaks at diffraction angles (2q) of 6.28±1°, 8.88±1°, 10.32±1°, 13.42±1°, 21.04±1°, 28.14±1°, 33.5±1° and 59.16±1°. When the above peaks are satisfied, the metal hydroxide composite can be provided in the form of a layered structure, and the layered structure can include a modified asymmetric layered structure. More specifically, methanol and / or water can be used as a solvent to form the layered structure. By adjusting the solvent ratio, an asymmetric layered structure and / or a modified layered structure can be formed, and the formed asymmetric layered structure and / or modified layered structure can be a layered structure having multiple layers, for example, 2 to 5 layers.

[0079] In the present invention, the multi-layered hydroxide structure can have a powder X-ray diffraction pattern with peaks at diffraction angles (2θ) of 6.02±1°, 8.5±1°, 10.4±1°, 13.38±1°, 21.02±1°, 26.42±1°, 27.88±1°, 33.52±1°, and 59.1±1°. When the above peaks are satisfied, the metal hydroxide composite can be provided in the form of a layered structure, and the layered structure can include a modified asymmetric layered structure. More specifically, propanol and / or water can be used as a solvent to form the layered structure. The propanol is preferably n-propanol.

[0080] In the present invention, the multi-layered hydroxide structure can have a powder X-ray diffraction pattern with peaks at diffraction angles (2θ) of 6.22±1°, 10.5±1°, and 13.36±1°, 21.04±1°, 20.94±1°, 33.46±1°, and 59.28±1°. When the above peaks are satisfied, the metal hydroxide composite can be provided in the form of a layered structure, and the layered structure can include a modified asymmetric layered structure. More specifically, butanol and / or water can be used as a solvent to form the layered structure. The butanol is preferably n-butanol.

[0081] More specifically, the multi-layered hydroxide structure can have a powder X-ray diffraction pattern with peaks at diffraction angles (2θ) of 5.96±0.5°, 8.5±0.5°, 10.36±0.5°, 13.36±0.5°, 19±0.5°, 20.84±0.5°, 21.7±0.5°, 26.34±0.5°, 37.68±0.5°, 31.48±0.5°, 33.78±0.5°, 34.88±0.5°, and 59.3±0.5°. When the above peaks are satisfied, the metal hydroxide composite can be provided in the form of a layered structure, and the layered structure can include a modified asymmetric layered structure and / or a modified layered structure. More specifically, ethanol and / or water can be used as a solvent to form the layered structure.

[0082] In the present invention, the multi-layered hydroxide structure can have a powder X-ray diffraction pattern with peaks at diffraction angles (2θ) of 6.28±0.5°, 8.88±0.5°, 10.32±0.5°, 13.42±0.5°, 21.04±0.5°, 28.14±0.5°, 33.5±0.5°, and 59.16±0.5°. When the above peaks are satisfied, the metal hydroxide composite can be provided in the form of a layered structure, and the layered structure can include a modified asymmetric layered structure and / or a modified layered structure. More specifically, methanol and / or water can be used as a solvent to form the layered structure.

[0083] In the present invention, the multi-layered hydroxide structure can have a powder X-ray diffraction pattern having peak values at diffraction angles (2θ) of 6.02±0.5°, 8.5±0.5°, 10.4±0.5°, 13.38±0.5°, 21.02±0.5°, 26.42±0.5°, 27.88±0.5°, 33.52±0.5°, and 59.1±0.5°. When the above peak values are satisfied, the metal hydroxide composite can be provided in the form of a layered structure, and the layered structure can include a modified asymmetric layered structure and / or a modified layered structure. More specifically, propanol and / or water can be used as a solvent to form the layered structure. The propanol is preferably n-propanol.

[0084] In the present invention, the multi-layered hydroxide structure can have a powder X-ray diffraction pattern having peak values at diffraction angles (2θ) of 6.22±0.5°, 10.5±0.5°, 13.36±0.5°, 21.04±0.5°, 20.94±0.5°, 33.46±0.5°, and 59.28±1°. When the above peak values are satisfied, the metal hydroxide composite can be provided in the form of a layered structure, and the layered structure can include a modified asymmetric layered structure and / or a modified layered structure. More specifically, butanol and / or water can be used as a solvent to form the layered structure. The butanol is preferably n-butanol.

[0085] The present invention relates to a metal hydroxide composite obtained by subjecting an active ingredient and a metal hydroxide structure precursor to co-precipitation through a precipitation reaction using ethanol and water.

[0086] More specifically, the metal hydroxide structure of the present invention can induce a precipitation reaction using a precursor.

[0087] In the present invention, specific examples of the metal hydroxide structure precursor that can cause the precipitation reaction preferably include ZnO, ZnSO4, ZnCl2, ZnCO3, Zn(NO3)2 6H2O, Zn(CH3COO)2, CaO, CaCl2, Ca(NO3)2 6H2O, CaSO4, CaCO3, Ca(OH)2, MgO, MgCl2, MgSO4, Mg(NO3)2 6H2O, CuO, Cu(NO3)2 6H2O, CuCl2, CuSO4, Co(NO3)2, CoCO3, CoCl2, Co(OH)2, CoSO4, Co(CH3COO)2, NiO, NiCl2, Ni(NO3)2, NiSO4, NiCO3, Ni(OH)2, FeO, FeCl3, Fe(NO3)3, FeSO4, FeCO3, and Fe(OH)2, and can preferably include ZnO, ZnSO4, ZnCl2, ZnCO3, Zn(NO3)2 6H2O, and Zn(CH3COO)2.

[0088] The alcohol can be at least one selected from the group consisting of methanol, ethanol, propanol, and butanol. The propanol and / or butanol can be n-propanol and / or n-butanol. In the co-precipitation step, the ratio of the alcohol and water can be 1:9 to 9:1, preferably 2.4:7.6 to 6.2:3.8. When the ratio of the alcohol and water falls within the above range, a modified multi-layer structure is formed, which is desirable.

[0089] The present invention relates to a method of manufacturing a metal hydroxide composite, which includes subjecting an active ingredient and a metal hydroxide structure to co-precipitation by using a precipitation reaction using an alcohol and water. The alcohol can be at least one selected from the group consisting of methanol, ethanol, propanol, and butanol. Herein, it is preferable that the propanol is n-propanol and the butanol is n-butanol.

[0090] More specifically, the method of the present invention includes forming a metal hydroxide structure precursor using a precursor, preparing an active ingredient solution by dissolving an active ingredient using an alcohol and water, and subjecting the metal hydroxide structure precursor and the active ingredient solution to a precipitation reaction.

[0091] In the present invention, when the active ingredient and the metal hydroxide are precipitated using the alcohol and water, the individual active ingredient can be introduced into the metal hydroxide structure without an additional process and / or step, thus enabling the active ingredient to be stabilized. In addition, using the alcohol and water as the solvent enables the number of process steps to be reduced, and the total process time of the metal hydroxide composite to be manufactured in the present invention is shortened to the range of 3 to 5 hours, preferably 3 hours and 30 minutes to 4 hours and 30 minutes. The conventional process for producing the same material takes 10 to 12 hours, which means that a significant effect of shortening the process time can be provided. In addition, at least 5 to 15% of the effect of further adding the active ingredient to the metal hydroxide composite can be achieved compared to the conventional amount, based on a rational formula. Herein, the alcohol can be at least one selected from the group consisting of methanol, ethanol, propanol, and butanol, and ethanol is preferably used because the active ingredient can be excessively and very effectively included in the metal hydroxide composite.

[0092] More specifically, the use of the alcohol during the synthesis process can prevent the deterioration (oxidation, browning, etc.) of the active substance, prevent the clogging of the ultrafiltration (UF) filter used to wash the obtained slurry after the synthesis, and prevent the deterioration of the material during the three times of UF washing process. In addition, the drying time can be reduced compared to the case of using only the water phase after washing and drying using a spray dryer, and since drying is performed at 65 to 75°C, preferably 70°C (when using water, drying is performed at 95°C or higher), the deterioration of the active substance can be prevented. In particular, it is preferable to use ethanol as the alcohol, so that the effects of the above processes can be very effectively achieved.

[0093] Mode for Invention

[0094] In the present invention, the basic principle of the synthesis of the metal hydroxide is a precipitation reaction. The precipitation reaction can be determined based on the following Equation 1, and is related to the solubility product of the following Equation 1. With regard to the generation of the precipitate, when the concentration product of the ions is equal to the solubility product of the precipitate, a saturated solution can be generated, and when the concentration product of the ions exceeds the solubility product of the precipitate, precipitation can occur beyond the saturated concentration. When the concentration product of the ions required for the precipitation is less than the solubility product of the precipitate, an unsaturated solution can be generated, and thus precipitation can not occur. Therefore, in order to form the precipitate, a predetermined amount or more of the precursor must be used so as to exceed the solubility product of the precipitate. However, if the amount of the precursor is too large, attention must be paid because problems such as the re-dissolution of the precipitate can occur due to the influence of the counter ion or the formation of a complex. Details of the precipitation reaction of the present application are shown in Figures 15 to 17 .

[0095] [Equation 1]

[0096] [Ksp] = [Zn 2+ ][OH - ] 2

[0097] Solubility product can be represented by Zn 2+ (aq), Zn(OH) + (aq), Zn(OH)2(aq), Zn(OH) 3- (aq), and Zn(OH)4 2- (aq) and based on the equilibrium or equilibrium constant shown, shows correlation with solubility data, thus making it easy to correlate the listed species with other species. Figure 15

[0098] Synthesis Example 1 : Preparation 1 of multilayer metal hydroxide crystals with Vitamin C (ascorbic acid) intercalated between the layers of the layered metal salt Synthesis Example 2: Preparation 2 of multilayer metal hydroxide crystals with Vitamin C intercalated between the layers of the layered metal salt

[0099] ZnO and 1197 mL of deionized water free from carbonate ions (CO3 2- ) were stirred in a main reaction tank (400 rpm) at room temperature under nitrogen atmosphere, and a concentrated hydrochloric acid solution was slowly added dropwise thereto so as to titrate the pH to the range of 0.5 to 1. 26 g of vitamin C was dissolved in 230 mL of deionized water and 175 mL of ethanol in a subsidiary tank 1, and the vitamin C solution of the subsidiary tank 1 was added to the main tank, which was stirred for 30 min under nitrogen atmosphere. While maintaining the stirring rate (700 rpm) of the main tank, 5M aqueous sodium hydroxide solution of subsidiary tank 2 was added thereto so as to titrate the pH to the range of about 6.5 to 7.5, and then a precipitation reaction was initiated for 3 hours under stirring (700 rpm). After the reaction, the pH was measured to be 6.5-7.0, and after the termination of the reaction, a white slurry precipitate was obtained. The precipitate was subjected to centrifugation and washed three times using ethanol (50%), thereby removing unreacted salt and vitamin C. After centrifugation, the precipitate separated from the supernatant was dried in a hot air dryer (inlet temperature: 70°C) using ethanol (95%), thereby obtaining 35.4 g of pale beige vitamin C-zinc hydroxide (yield 58.8%, ascorbic acid content 42.24%).{[Zn3(OH)4] B [Zn2(OH)3] S}{(ascorbic acid)2}·2(H2O)

[0100] Analytical methods and evaluation criteria Figure 1

[0101] ZnO, 549 mL of deionized water free from carbonate ions (CO3 2- ​) deionized water and 628mL of ethanol (>98%) are stirred (400rpm) in the main reaction tank, in a nitrogen atmosphere, at room temperature, concentrated hydrochloric acid solution is slowly added dropwise thereto so that its pH is titrated to the range of 0.5 to 1. 26g vitamin C is dissolved in 230mL deionized water and 175mL of ethanol in auxiliary tank 1, and the vitamin C solution of auxiliary tank 1 is added to the main tank and stirred for 30min in a nitrogen atmosphere. While maintaining the stirring speed (700rpm) of the main tank, the 5M sodium hydroxide aqueous solution of auxiliary tank 2 is added thereto so that its pH is titrated to the range of about 6.5 to 7.5, and then precipitation reaction is initiated for 3 hours under stirring (700rpm). After the reaction, measuring pH is 6.5 to 7.0, and after the reaction terminates, a white mud precipitate is obtained. The precipitate is centrifuged and washed three times with ethanol (50%) to remove unreacted salt and vitamin C. After centrifugation, the precipitate separated from the supernatant was dried using ethanol (95%) in a hot air dryer (inlet temperature: 70° C.) to obtain 37.6 g of vitamin C-zinc hydroxide having a pale ivory color close to white (yield 62.4%, ascorbic acid content 45.19%).

[0102] {[Zn3(OH)4] B [Zn1(OH)2] S}{(ascorbic acid)2}·3(H2O)

[0103] Synthesis Example 3: Preparation of multilayer metal hydroxide crystals 3, wherein vitamin C is inserted between the layers of the layered metal salt

[0104] 30g ZnO, 254mL carbonate ion-free (CO3 2-) deionized water and 943mL of ethanol (>98%) are stirred (400rpm) in the main reaction tank, in a nitrogen atmosphere, at room temperature, concentrated hydrochloric acid solution is slowly added dropwise thereto so that its pH is titrated to the scope of 0.5 to 1. 26g vitamin C is dissolved in 230mL deionized water and 175mL of ethanol in auxiliary tank 1, and vitamin C solution is added to the main tank and stirred for 30min in a nitrogen atmosphere. While maintaining the stirring speed (700rpm) of the main tank, the 5M sodium hydroxide aqueous solution of auxiliary tank 2 is added thereto so that its pH is titrated to the scope of about 6.5 to 7.5, and then precipitation reaction is initiated for 3 hours under stirring (700rpm). After the reaction, measuring pH is 6.5 to 7.0, and after the reaction terminates, a white mud precipitate is obtained. The precipitate is centrifuged and washed three times with ethanol (50%) to remove unreacted salt and vitamin C. After centrifugation, the precipitate separated from the supernatant was dried using ethanol (95%) in a hot air dryer (inlet temperature: 70° C.), thereby obtaining 39.5 g of a product having a light gray color close to white (yield 65.6%, ascorbic acid content 46.35%). B [Zn1(OH)1] S}{(ascorbic acid)2}·3(H2O)

[0105] Synthesis Example 4: Preparation of multilayer metal hydroxide crystals 4, wherein vitamin C is inserted between the layers of the layered metal salt

[0106] 30g ZnO, 68mL carbonate ion-free (CO3 2- ) deionized water and 1129mL of ethanol (>98%) are stirred (400rpm) in the main reaction tank, in a nitrogen atmosphere, at room temperature, concentrated hydrochloric acid solution is slowly added dropwise thereto so that its pH is titrated to the scope of 0.5 to 1. 26g vitamin C is dissolved in 50mL deionized water and 355mL of ethanol in auxiliary tank 1, and vitamin C solution is added to the main tank and stirred for 30min in a nitrogen atmosphere. While maintaining the stirring speed (700rpm) of the main tank, the 5M sodium hydroxide aqueous solution of auxiliary tank 2 is added thereto so that its pH is titrated to the scope of about 6.5 to 7.5, and then precipitation reaction is initiated for 3 hours under stirring (700rpm). After the reaction, measuring pH is 6.5 to 7.0, and after the reaction terminates, a white mud precipitate is obtained. The precipitate is centrifuged and washed three times with ethanol (50%) to remove unreacted salt and vitamin C. After centrifugation, the precipitate separated from the supernatant was dried using ethanol (95%) in a hot air dryer (inlet temperature: 70° C.), thereby obtaining 40.9 g of a product having a light gray color close to white (yield 67.8%, ascorbic acid content 48.84%).B [Zn1(OH)1] S}{(ascorbic acid)2}·2(H2O)

[0107] Figure 1

[0108] To determine the structure of the multilayered metal hydroxide, analysis was performed using the following analyzer.

[0109] Analysis method 1: Powder-X-ray diffraction pattern

[0110] - Instrument: Powder X-ray diffraction (PXRD)

[0111] X-ray diffractometer (D / MAX PRINT 2200-Ultima, Japan, Rigaku)

[0112] Cu-Kα radiation (λ = 1.5418 )

[0113] Tube voltage: 40 kV, current: 30 mA

[0114] As the X-ray diffractometer, D / MAX PRINT 2200-Ultima available from Rigaku (Japan) was used. The anode generating X-rays was formed of Cu metal, and measurement was performed using Kα rays (λ = 1.5418 ) at 3 to 70° of 2θ, 0.02° / 0.2 sec scanning speed, and the divergence slit, the scattering slit, and the receiving slit were 0.1, 1, and 1 mm, respectively. The tube voltage was 40 kV, and the current was 30 mA.

[0115] Evaluation criteria 1

[0116] The one-dimensional (1D) electron density of the z-axis of PXRD was calculated using the following Equation 2.

[0117] [Equation 2]

[0118]

[0119] The powder obtained by synthesis was comparatively analyzed using the XRD pattern, and the interlayer distance was calculated using Bragg’s equation (Equation 3 below). For the peak located at the front, which indicates the interlayer distance including the distance of the metal hydroxide layer and the anion layer synthesized, it was regarded as the main interlayer distance.

[0120] [Equation 3]

[0121] nλ = 2dsinθ

[0122] (λ = X-ray wavelength, d = lattice spacing, θ = angle of incidence)

[0123] Analytical method 2: HPLC analysis

[0124] - Instrument: High Performance Liquid Chromatography (HPLC) analysis

[0125] Agilent 1100 series (Agilent Technologies, USA)

[0126] UV detector (λmax = 240)

[0127] Zorbax C18 column (4.6 mm x 150 mm, 5 μm, Agilent Technologies, USA)

[0128] Flow rate: 0.65 ml / min

[0129] Injection volume: 10 μl

[0130] Column temperature: 35 °C

[0131] For HPLC analysis, Agilent 1100 series (Agilent Technologies, USA) was used. λmax was measured at 240 nm and the measurement was performed using Zorbax C18 column (4.6 mm x 150 mm, 5 μm, Agilent Technologies, USA) at 0.65 ml / min, 10 μl injection volume, 35 °C column temperature.

[0132] The mobile phase buffer included 0.1 % trifluoroacetic acid (ReagentPlus , 99 %) and used acetonitrile (anhydrous, 99.8 %) and deionized water in a volume ratio of 2:8.

[0133] For sample treatment, 40 mg of sample was mixed with 100 ml of buffer solvent, sonicated for 10 minutes and then stirred rapidly for 10 minutes. The resulting solution was filtered using a nylon syringe filter (0.2 μm pore size) and the sample was measured.

[0134] Experimental Example 1

[0135] The graphs analyzed by the above analytical method are shown in Examples 1-4 and in Table 1 below.

[0136] Method of preparing reference (Zn5(OH)8(ascorbic acid)2-2H2O)

[0137] 6 g of Zn(N03)2-6H20 and 1.42 g of vitamin C were dissolved in a carbonate ion (C03 2-) to pH about 6 to 7 using 0.2 M NaOH, and then allowed to react for 12 hours to obtain a precipitate of the basic salt of Vitamin C-Zinc. Then, the titrated solution was subjected to centrifugation to separate the supernatant and washed 6 times to remove the unreacted salt and Vitamin C, thereby obtaining 2.9 g (yield 65%, Vitamin C content: 39%).

[0138] Method of preparing MZLA

[0139] {[Zn3(OH)4] B [Zn1(OH)2] S}{(ascorbate)2} 3(H2O)

[0140] The MZLA of the present invention was prepared in the same manner as in Synthesis Example 2.

[0141] Method of preparing the basic salt of zinc (Zn5(OH)8(NO3)2 2H2O)

[0142] 5 g of Zn(NO3)2 6H2O was dissolved in deionized water free of carbonate ions (CO3 2- ) and titrated to pH about 6 to 7 using 0.2 M NaOH to obtain a precipitate of the basic salt of zinc. Then, the titrated solution was subjected to centrifugation and washing, thus removing the unreacted salt, thereby obtaining 2.6 g of white powder (yield: 70%).

[0143] [Table 1]

[0144]

[0145] Table. Powder XRD pattern of the corresponding basal spacing

[0146] As shown in the XRD pattern of Figure 2 , the physical structure change of the metal coordination sites in the metal sheet appears as a change in the charge in the basal area unit, and the number, size, binding strength, and orientation of the ascorbate are determined by the physical properties of the ascorbate, such as steric hindrance, electron density, ionic dipole moment, etc., which are coupled with the metal layer within the sheet to provide the interlayer height.

[0147] The interlayer distance of the metal hydroxide (MZLA) in this experiment was 14.82 , 10.4 , and 8.53 The d values ​​of the three peak positions below 15° are different from each other, and considering the ratio of the interlayer distance equation (Bragg equation) to the stacking order of the c-axis, different types of stacking are obtained. This is because The reflections form different base layers (basal plane spacings) and separate peaks are generated from different structures. In addition, considering the layer thickness (brucite layer thickness, 7.4 ) and ascorbate ion length (4.9 ), 8.5°(10.394 ) is further reduced by about 4.4° compared to 5.96°. , and 10.36°(8.6419 ) decreased by 6.29 , and due to the NO of ZBS 3- The ions show a specific interlayer distance (9.82 ) is small, indicating that the metal coordination sites formed by the metal layer and ascorbate have changed, resulting in a smaller interlayer distance.

[0148] When compared with the conventional hydroxide structure, the crystallinity of the peak representing the spacing of the layered structure increases, and new peaks are generated. The interlayer spacing of conventional metal hydroxides is 14.67 , but a new peak was generated in MZLA (modified zinc layer ascorbate) with a calculated interlayer spacing of 14.81 , 10.39 and 8.53 , resulting in a change in the diffraction pattern and an increase in crystallinity.

[0149] Experimental Example 2: XRD pattern and measurement of vitamin C content depending on ethanol and water content

[0150] Figure 3

[0151] Using the amounts shown in Table 2 below, Example 1, Example 2, Example 3, and Example 4 were synthesized in the same manner as Synthesis Example 1, Synthesis Example 2, Synthesis Example 3, and Synthesis Example 4, respectively.

[0152] [Table 2]

[0153]

[0154] As shown in Table 2 and Figure 3As shown, depending on the ratio of ethanol and deionized water at the time of synthesis, the crystallinity of the peaks varies and the interlayer distance changes. When the ethanol ratio is low (Example 1, (a)), the diffraction pattern of ascorbate salt appears widely near 5-6°, and as the ethanol ratio gradually increases, the crystallinity of the peaks increases, and a new diffraction pattern appears, thus generating distinct peaks near 8° and 10°.

[0155] In addition, as the ethanol ratio increases, the amount of ascorbate salt of the metal hydroxide gradually increases to 44.24%, 45.19%, and 48.84%, and the color of the powder also shows a pale ivory color close to white.

[0156] Experimental Example 3: Analysis of XRD changes depending on ① EtOH:H2O = 85:15 ratio, ② EtOH:H2O = 10:90 ratio, ③ H2O = 100 (reference), and ④ ZBS-NO3

[0157] The Zn site constituting the tetrahedral structure in the layered structure is in a positively charged unstable form, and reacts with an ionic molecule in order to balance the charge valance corresponding thereto, and the water molecule combined to the Zn site by a dipole moment is combined with anions by using hydrogen bonding and ionic dipole interaction to form a metal layer. Herein, when EtOH is used in a predetermined ratio or more, competition between solvent molecules having a small dipole moment promotes the instability of the water molecule combined to the Zn site, causing a change in the structural position of Zn constituting the tetrahedral structure in the ZBS layer. The ascorbate salt of ZBS-ascorbate salt exists by the combination of the water molecule combined to Zn and ascorbic acid (dehydrated form), and the structural instability generated by ethanol affects the ascorbate salt combined to the water molecule.

[0158] Figures 8 to 10 This tendency is confirmed in the middle, and for the same reason as described above, when ethanol of a predetermined ratio is used, a twisted asymmetric layered structure (turbostatic structure) of nanosheet shape is formed in order to maintain a structurally stable form, and the peaks near 33° and 58° appear in a broad humped form ). As the amount of ethanol increases, the position of the anion in the layer sheet changes in order to maintain a more structurally stable form, and peaks representing the layer distance below 15° appear and change. In addition, when the asymmetric structure disappears, the peaks near 33° and 58° disappear, and a multi-layer type layered structure ) appears.

[0159] The asymmetric layered structure (turbostatic structure) of the diffraction pattern at 30°<θ<40° and 55°<θ<65° is not found in ZBS-NO3 ), but this asymmetric lamellar structure was found in the structure of other anions contained in ZBS. As Examples 5-7 , did not occur in the case of the conventional ZBS-ascorbic acid, but it was a physical phenomenon artificially created by controlling the ratio of EtOH, showing that the structure and form were different from the XRD peaks indicated by the reference.

[0160] Experimental Example 4: Scanning Electron Microscope of Metal Hydroxide Synthesized in Example 2

[0161] Figure 4 Scanning Electron Microscope image (SEM) of the metal hydroxide prepared in Example 2. The metal hydroxide had an average particle size of 10 to 20 μm, and had a spherical shape. The size thereof was different from the size of the reference, and the reference was known to have a size of about 200 to 300 nm or 0.5 μm or less.

[0162] Experimental Example 5: XRD pattern depending on ethanol and water content and measurement of vitamin C content

[0163] Figure 5

[0164] Example 5, Example 6, and Example 7 were synthesized in the same manner as in Synthetic Example 2, Synthetic Example 3, and Synthetic Example 4, respectively, except that the amounts shown in Table 3 below were applied.

[0165] The specific XRD values of Examples 5 to 7 are shown in Example 8(a) .

[0166] [Table 3]

[0167]

[0168] Experimental Example 6: XRD pattern depending on methanol and water content and measurement of vitamin C content

[0169] Examples 8 to 10

[0170] Example 8, Example 9, and Example 10 were synthesized in the same manner as in Synthetic Example 2, Synthetic Example 3, and Synthetic Example 4, respectively, except that methanol was used instead of ethanol, and the amounts shown in Table 4 below were applied.

[0171] The float XRD values of Examples 8 to 10 are shown in Example 9(b) and Table 5 below.

[0172] [Table 4]

[0173] ​

[0174] [Table 5]

[0175] Example 10(c) 6.48 33.42 59.3 Figure 6 6.34 8.56 10.4 13.52 21.14 28.2 33.52 59.08 Example 11(a) 6.28 8.62 10.28 13.42 21 28.06 33.5 59.16

[0176] Experimental Example 7: XRD patterns and measurement of vitamin C content depending on the content of propanol and water

[0177] Examples 11 to 13

[0178] Examples 11, 12, and 13 were synthesized in the same manner as in Synthetic Example 2, Synthetic Example 3, and Synthetic Example 4, respectively, except that n-propanol was used instead of ethanol, and the amounts shown in Table 6 below were applied.

[0179] Specific XRD values for Examples 11-13 are shown in Example 12(b) .

[0180] [Table 6]

[0181]

[0182] [Table 7]

[0183] Example 13(c) 6.04 10.38 13.2 20.78 33.38 59.2 Figure 7 6.12 8.44 10.38 13.36 21.06 26.4 27.88 33.46 59.24 Example 11(a) 6.02 8.54 10.38 13.32 20.96 26.5 27.88 33.56 59.1

[0184] Experimental Example 8: XRD patterns and measurement of vitamin C content depending on the content of butanol and water

[0185] Examples 14 to 17

[0186] Examples 14, 15, and 16 were synthesized in the same manner as in Synthetic Example 2, Synthetic Example 3, and Synthetic Example 4, respectively, except that n-butanol was used instead of ethanol, and the amounts shown in Table 8 below were applied.

[0187] Specific XRD values for Examples 14-16 are shown in Example 12(b) .

[0188] [Table 8]

[0189]

[0190] [Table 9]

[0191] Example 13(c) 6.5 13.54 20.42 33.44 59.22 Figure 12 6.4 13.5 20.92 33.32 59.22 Figure 12 6.38 10.44 13.4 20.94 33.4 59.24

[0192] Experimental Example 9: Collagen expression effect of Example 3

[0193] - Experimental Methods

[0194] The test of Example 3 (hereinafter referred to as "JL-101 powder") was performed with respect to the increase in collagen production in vitro, using normal human dermal fibroblasts (HDF). In order to set the test solution concentration of the sample, HDF were treated with the sample material for 24 hours, and the concentration range in which no cytotoxicity was observed was confirmed by a cytotoxicity test. HDF were treated with JL-101 powder in the same manner at the set experimental solution concentration, and the expression levels of COL1A1 and COL3A1 mRNA, which are collagen genes, were compared and analyzed using real-time PCR (RT-PCR).

[0195] - Cell line selection and cell culture

[0196] For this test, normal human dermal fibroblasts (nHDF; Lonza, Basel, Switzerland) were used. These cells were cultured in a 5% CO2 incubator at 37°C using fibroblast basal medium (cc-3181, Lonza, Walkersville, MD, USA) and FGM-7 supplement (cc-4126, Lonza, Walkersville, MD, USA). TM - 2 Single Quots TM (hGFG, insulin, FBS, and gentamicin / amphotericin B; cc-4126, Lonza, Walkersville, MD, USA) at 5% CO2 incubator.

[0197] - Test material

[0198] As the test material, "JL-101" was provided in the form of a powder, and was dissolved in phosphate buffered saline (PBS; Biosesang, Seongnam, Korea) and used for cell treatment. As a positive control, transforming growth factor beta (TGF-β; T7039, Sigma-Aldrich, MO, USA) was dissolved in PBS and used for cell treatment.

[0199] - Test material concentration setting

[0200] Human dermal fibroblasts were seeded at 4 x 10 3Cells were seeded at 2,000 cells / well, incubated for 24 hours, treated with test materials at an appropriate concentration, and further incubated for 24 hours. Subsequently, the cells were treated with a WST-1 test solution at 10% of the medium volume, and allowed to react for an additional 0.5 to 1 hour at 37°C, after which the absorbance was measured at 450 nm using an iMark microplate reader (Bio-Rad, Hercules, CA, USA). The reference absorbance was measured at 650 nm to calibrate the result values. The result values were represented by the mean ± standard deviation based on three independent experiments. P <.05 was confirmed by Student’s t-test to show the significance of the result values. Based on the result values, the concentration range in which the cell activity was not decreased due to treatment with the test materials compared to the control was set as the test solution concentration in the subsequent experiment.

[0201] - Test of expression level of collagen genes

[0202] The expression level of collagen genes was determined using quantitative real-time PCR (qRT-PCR) (Peter, 2008) by measuring the expression level of the COL1A1 gene, which is a representative component of type 1 collagen, and the COL3A1 gene, which is a representative component of type 3 collagen. Human dermal fibroblasts were seeded at 2 x 10 5 Cells were seeded at 2,000 cells / well, incubated for 24 hours, treated with test materials at an appropriate concentration, and further incubated for 24 hours. Subsequently, the cells were treated with a WST-1 test solution at 10% of the medium volume, and allowed to react for an additional 0.5 to 1 hour at 37°C, after which the absorbance was measured at 450 nm using an iMark microplate reader (Bio-Rad, Hercules, CA, USA). The reference absorbance was measured at 650 nm to calibrate the result values. The result values were represented by the mean ± standard deviation based on three independent experiments. P <.05 was confirmed by Student’s t-test to show the significance of the result values. Based on the result values, the concentration range in which the cell activity was not decreased due to treatment with the test materials compared to the control was set as the test solution concentration in the subsequent experiment. The total mRNA was measured using a MaestroNano micro-volume spectrophotometer (Maestrogen, Las Vegas, NV, USA), and only total mRNA with a high purity of 2.0 or more was selected.

[0203] Total mRNA was replaced with cDNA using M-MLV reverse transcriptase (Promega) and used for qRT-PCR. qRT-PCR was performed using HOT FIREPol EvaGreen PCR Mix Plus (Solis BioDyne, Estonia) and the expression of the corresponding genes was analyzed using Line-Gene K software (Bio-Rad Laboratories, Inc., Hercules, CA, USA). As a positive control for collagen and collagenase mRNA content measurement test, TGF-β was used (Chung et al. 1997). PCR was performed by denaturation at 94°C for 5 min and 40 cycles of denaturation (94°C, 30 sec), annealing (60°C, 30 sec) and polymerization (72°C, 30 sec). Changes in gene expression were measured by comparison with the expression level of the β-ACTIN control gene. The CT values of COL1A1 and COL3A1 were normalized to the CT value of β-ACTIN and the relative expression levels of COL1A1 and COL3A1 were calculated by the 2-ΔΔCt method. Resultant values are represented by the mean ± standard deviation based on three independent experiments. P<.05 was confirmed by Student's t-test to show the significance of the resultant values. Primer information for qRT-PCR is shown in Table 10 below. T 2- 2- [Table 10]

[0205]

[0206] - Setting of test solution concentration of test material

[0207] The test solution concentration of "JL-101 powder" as a test material was set using the WST test. Human dermal fibroblasts were treated with "JL-101 powder" material at a concentration range of 0-500 μg / mL for 24 hours, and the cell activity thereof was measured using the WST-1 test. The results thereof are shown in Table 11 below. Figure 13 When the concentration of "JL-101 powder" material used for cell treatment was 100 μg / mL or less (10, 20, 50 and 100 μg / mL), the cell activity did not show a significant difference compared to the negative control. However, when the concentration thereof was 200 μg / mL or more (200, 300, 400 and 500 μg / mL), the cell activity was reduced by 5% or more compared to the negative control. Therefore, in the subsequent potency test, the concentration of "JL-101 powder" was set to a test solution concentration of 100 μg / mL or less. Figure 13

[0208] - Measurement of collagen type 1 mRNA expression level ​​

[0209] By measuring the changes in the expression levels of COL1A1 mRNA and COL3A1 mRNA (which are the type 1 collagen and type 3 collagen genes mainly present in the skin), it was evaluated whether the expression of collagen in human dermal fibroblasts treated with the test material "JL-101 powder" increased or decreased, and the expression level of COL1A1 mRNA treated with "JL-101 powder" was first measured. Human dermal fibroblasts were treated with the test material "JL-101 powder" at a maximum test solution concentration (set by the above-mentioned toxicity test) of less than 100 μg / mL (10, 20, 50 and 100 μg / mL), and then cultured for 24 hours, and then total RNA was extracted and cDNA was synthesized. Subsequently, the expression level of COL1A1 mRNA was measured by qRT-PCR using the COL1A1 primer set and β-ACTIN primer set shown in Table 11. TGF-β was used as a positive control to improve the reliability of the test results. As Figure 14 As shown, COL1A1 mRNA expression increased in a concentration-dependent manner after treatment with "JL-101 Powder" at concentrations below 100 μg / mL. Specifically, COL1A1 mRNA expression increased by 45.88±6.79% compared to the negative control after treatment with 100 μg / mL. Based on the results of treatment with the positive control TGF-β (5.0 ng / ml), COL1A1 mRNA expression increased by 219.65±7.25% compared to the negative control ( Figure 14 ).

[0210] Measurement of type 3 collagen mRMA expression levels

[0211] like Figure 18 As shown in Table 1, based on the expression analysis of COL1A1 mRNA treated with "JL-101 powder", the expression of COL3A1 mRNA treated with "JL-101 powder" was analyzed. Human dermal fibroblasts were treated with the test material "JL-101 powder" within the test solution concentration range (10, 20, 50 and 100 μg / mL) set in the above-mentioned type 1 collagen mRNA expression level measurement test and cultured for 24 hours. Thereafter, total RNA was extracted and cDNA was synthesized. The expression level of COL3A1 mRNA was measured by qRT-PCR using the COL3A1 primer set and β-ACTIN primer set shown in Table 1. TGF-β was used as a positive control to improve the reliability of the test results. As ​As shown, the expression of COL3A1 mRNA was increased in a concentration-dependent manner after treatment with 0 to 100 pg / mL of "JL-101 powder". Specifically, the expression of COL3A1 mRNA was increased by 100.98 ± 5.49% after treatment with 100 pg / mL of "JL-101 powder" compared to the negative control. Based on the results of treatment with the positive control TGF-β (5.0 ng / ml), the expression of COL3A1 mRNA was increased by 22.93 ± 6.21% compared to the negative control.

[0212] - Measurement results

[0213] The Korea Dermatology Research Institute performed a potency test by testing the increase in collagen production in human dermal fibroblasts by the test material "JL-101 powder" at the request of C&Pharm.

[0214] Based on the results of the analysis of human dermal fibroblast activity by the test material "JL-101 powder" provided by C&Pharm, the cell activity did not decrease in the case of 24 hours of culture after treatment with a concentration of 100 pg / mL or less (10, 20, 50, and 100 pg / mL) of "JL-101 powder" compared to the negative control. In the potency test for increasing collagen expression, the expression of COL1A1 mRNA was significantly increased after treatment with 20, 50, and 100 pg / mL of "JL-101 powder" compared to the negative control. Specifically, the expression of COL1A1 mRNA after treatment with 100 pg / mL of "JL-101 powder" was increased by 45.88 ± 6.79% compared to the untreated negative control. Based on the results of human dermal fibroblasts treated with the positive control TGF-β (5.0 ng / ml) at a concentration of 5.0 ng / mL, the expression of COL1A1 mRNA was increased by 22.93 ± 7.25% compared to the negative control. The expression of COL3A1 mRNA was increased in a concentration-dependent manner after treatment with 100 pg / mL or less of "JL-101 powder". Specifically, the expression of COL3A1 mRNA after treatment with 100 pg / mL of "JL-101 powder" was increased by 100.98 ± 5.49% compared to the untreated negative control. Based on the results of human dermal fibroblasts treated with the positive control TGF-β at a concentration of 5.0 ng / mL, the expression of COL3A1 mRNA was increased by 22.93 ± 6.21% compared to the negative control.

[0215] Thus, the expression of COL1A1 mRNA was increased by 45.88 ± 6.79% and the expression of COL3A1 mRNA was increased by 100.98 ± 5.49% when human dermal fibroblasts were treated with the test material supplied by C&Pharm, as compared to the negative control. The collagen expression levels were 45.64% and 163.50% of the collagen expression levels of the positive control (TGF-β), respectively. Thus, it was determined that "JL-101 powder" increases the expression of collagen in human dermal fibroblasts.

[0216] Reference Examples 1 and 2

[0217] Preparation of Reference Example 1

[0218] EtOH 0.5:H2O 9.5

[0219] 3 g of ZnO and 148.3 mL of deionized water free of carbonate ions (CO3 2- ) were stirred (400 rpm) in a main reaction tank under a nitrogen atmosphere at room temperature, and a concentrated hydrochloric acid solution was slowly added dropwise thereto so as to titrate the pH to the range of 0.5 to 1. 2.6 g of vitamin C (ascorbic acid) was dissolved in 10 mL of deionized water and 8.69 mL of ethanol in a subsidiary tank 1, and the vitamin C solution of the subsidiary tank 1 was added to the main tank, which was stirred under a nitrogen atmosphere for 30 min. While maintaining the stirring rate (700 rpm) of the main tank, 1.6 M of an aqueous sodium hydroxide solution of a subsidiary tank 2 was added thereto so as to titrate the pH to the range of about 6.5 to 7.5, and then a precipitation reaction was induced under stirring (700 rpm) for 3 hours. After the reaction, the pH was measured to be 6.5 to 7.0, and after the termination of the reaction, a white mud precipitate was obtained. The precipitate was subjected to centrifugation and washing three times so as to remove unreacted salts and vitamin C. After centrifugation, the precipitate separated from the supernatant was cooled with liquid nitrogen and then freeze-dried, thereby obtaining 3.5 g of light beige vitamin C-zinc hydroxide (yield 58.1%, ascorbic acid content 38.66%).

[0220] Preparation of Reference Example 2

[0221] EtOH 9.5:H2O 0.5

[0222] 3 g of ZnO and 125.15 mL of deionized water free of carbonate ions (CO3 2-) was stirred (400 rpm) in a main reaction tank under a nitrogen atmosphere at room temperature, and a concentrated hydrochloric acid solution was slowly added dropwise thereto so as to titrate the pH to the range of 0.5 to 1. 2.6 g of vitamin C was dissolved in 1.84 mL of deionized water and 40 mL of ethanol in an auxiliary tank 1, and the vitamin C solution of the auxiliary tank 1 was added to the main tank, which was stirred under a nitrogen atmosphere for 30 min. While maintaining the stirring rate (700 rpm) of the main tank, an aqueous 1.6 M sodium hydroxide solution of auxiliary tank 2 was added thereto so as to titrate the pH to the range of 6.5 to 7.5, and then a precipitation reaction was initiated under stirring (700 rpm) for 3 hours. After the reaction, the pH was measured to be about 6.5 to 7.0, and after the termination of the reaction, a white mud precipitate was obtained. The precipitate was subjected to centrifugation and washed three times so as to remove unreacted salt and vitamin C. After centrifugation, the precipitate separated from the supernatant was cooled with liquid nitrogen and then lyophilized, thereby obtaining 3.41 g of light beige vitamin C-zinc hydroxide (yield 56.6%, ascorbic acid content 34.24%).

[0223] The thus-prepared Reference Examples 1 and 2 are shown in Table 11 below, and the specific XRD values of Reference Examples 1 and 2 are shown in ​

[0224] [Table 11]

[0225]

[0226] Industrial applicability

[0227] According to the present application, the metal hydroxide complex is configured so that the active ingredient is included in the modified multi-layered hydroxide structure, thereby exhibiting excellent sustained-release performance, and the effect of the delivered active ingredient is improved due to the stable inclusion of a high content of the active ingredient in the metal hydroxide complex. In addition, the complex stably includes unstable active ingredients, thereby enabling long-term storage of the active ingredient, and when it is delivered to the skin, irritation can be minimized, thereby providing irritation-free action and enabling excellent collagen expression effects to be exhibited.

[0228] In addition, the present application provides a method of preparing a metal hydroxide complex having excellent effects as described above.​

Claims

1. A metal hydroxide complex comprising: A multilayer hydroxide structure of the following Chemical Formula 1, comprising a base layer, a surface layer, and an active component: {[M x (OH) z ] B [M y (OH) w ] S }{(A n ) q }·m(H2O)[chemical formula I], Among them [M x (OH) z ] B represents the base layer of the multilayer hydroxide structure, and [M y (OH) w ] S represents the surface layer of the multilayer hydroxide structure, M is selected from Zn 2+ divalent metal cations, x is 0.6 to 3, y is greater than 0 and not more than 2, z is 1 to 5, w is greater than 0 and not more than 4, z+w is 1 to 9, q is 1 to 4, m is 0.1 to 10, n is the charge of A, and A represents an active ingredient, wherein A is an anionic compound containing ascorbic acid, The multilayered hydroxide structure has a powder X-ray diffraction pattern with peaks at diffraction angles (2θ) of 5.96±1°, 33.46±1°, and 59.3±1°.

2. The metal hydroxide complex according to claim 1, wherein The multilayer hydroxide structure includes 2 to 5 layers.

3. The metal hydroxide complex according to claim 1, wherein The metal hydroxide complex includes about 45% to about 46% ascorbic acid.

4. The metal hydroxide complex according to claim 1, wherein The metal hydroxide complex includes about 45.23% to about 46.1% ascorbic acid.

5. The metal hydroxide complex according to claim 1, wherein The metal hydroxide composite is in the form of a plurality of particles.

6. The metal hydroxide complex according to claim 5, wherein The plurality of particles are substantially spherical in shape.

7. The metal hydroxide complex according to claim 5, wherein The plurality of particles have an average particle size of about 10 μm to about 20 μm.

8. The metal hydroxide complex according to claim 1, wherein The metal hydroxide complex exhibits a relative cell viability of at least about 80% or more at a concentration of about 10 μg / ml to about 300 μg / ml.

9. The metal hydroxide complex according to claim 1, wherein The metal hydroxide complex exhibits a relative cell viability of at least about 90% or more at a concentration of about 10 μg / ml to about 300 μg / ml.

10. The metal hydroxide complex according to claim 1, wherein The metal hydroxide complex at a concentration of about 10 μg / ml to about 300 μg / ml increases expression of COL1A1, COL3A1, or a combination thereof.

11. A metal hydroxide complex comprising: A multilayer hydroxide structure of the following Chemical Formula 1, comprising a base layer, a surface layer, and an active component: {[M x (OH) z ] B [M y (OH) w ] S }{(A n ) q }·m(H2O)[Chemical Formula 1] Among them, [M x (OH) z ] B represents the base layer of the multilayer hydroxide structure, and [M y (OH) w ] S represents the surface layer of the multilayer hydroxide structure, M is selected from Zn 2+ divalent metal cations, x is 0.6 to 3, y is greater than 0 and not more than 2, z is 1 to 5, w is greater than 0 and not more than 4, z+w is 1 to 9, q is 1 to 4, m is 0.1 to 10, n is the charge of A, and A represents an active ingredient, wherein A is an anionic compound containing ascorbic acid, wherein the multilayer hydroxide structure has a powder-X-ray diffraction pattern with peaks at diffraction angles (2θ) of 5.96±1°, 8.5±1°, 10.36±1°, 13.36±1°, 19±1°, 20.84±1°, 21.7±1°, 26.34±1°, 37.68±1°, 31.48±1°, 33.78±1°, 34.88±1° and 59.3±1°.

12. The metal hydroxide complex according to claim 11, wherein The multilayer hydroxide structure includes 2 to 5 layers.

13. The metal hydroxide complex according to claim 11, wherein The metal hydroxide complex includes about 45% to about 46% ascorbic acid.

14. The metal hydroxide complex according to claim 11, wherein The metal hydroxide complex includes about 45.23% to about 46.1% ascorbic acid.

15. The metal hydroxide complex according to claim 11, wherein The metal hydroxide composite is in the form of a plurality of particles.

16. The metal hydroxide complex according to claim 15, wherein The plurality of particles are substantially spherical in shape.

17. The metal hydroxide complex according to claim 15, wherein The plurality of particles have an average particle size of about 10 μm to about 20 μm.

18. The metal hydroxide complex according to claim 11, wherein The metal hydroxide complex exhibits a relative cell viability of at least about 80% or more at a concentration of about 10 μg / ml to about 300 μg / ml.

19. The metal hydroxide complex according to claim 11, wherein The metal hydroxide complex exhibits a relative cell viability of at least about 90% or more at a concentration of about 10 μg / ml to about 300 μg / ml.

20. The metal hydroxide complex according to claim 11, wherein The metal hydroxide complex at a concentration of about 10 μg / ml to about 300 μg / ml increases expression of COL1A1, COL3A1, or a combination thereof.

21. A metal hydroxide complex comprising: A multilayer hydroxide structure of the following Chemical Formula 1, comprising a base layer, a surface layer, and an active component: {[M x (OH) z ] B [M y (OH) w ] S }{(A n ) q }·m(H2O)[Chemical Formula 1] Among them, [M x (OH) z ] B represents the base layer of the multilayer hydroxide structure, and [M y (OH) w ] S represents the surface layer of the multilayer hydroxide structure, M is selected from Zn 2+ divalent metal cations, x is 0.6 to 3, y is greater than 0 and not more than 2, z is 1 to 5, w is greater than 0 and not more than 4, z+w is 1 to 9, q is 1 to 4, m is 0.1 to 10, n is the charge of A, and A represents an active ingredient, wherein A is an anionic compound containing ascorbic acid, wherein the multilayer hydroxide structure has a powder-X-ray diffraction pattern having peaks at diffraction angles (2θ) of 6.28±1°, 8.88±1°, 10.32±1°, 13.42±1°, 21.04±1°, 28.14±1°, 33.5±1°, and 59.16±1°.

22. The metal hydroxide complex according to claim 21, wherein The multilayer hydroxide structure includes 2 to 5 layers.

23. The metal hydroxide complex according to claim 21, wherein The metal hydroxide complex includes about 46% to about 51% ascorbic acid.

24. The metal hydroxide complex according to claim 21, wherein The metal hydroxide complex includes about 46.11% to about 50.97% ascorbic acid.

25. The metal hydroxide complex according to claim 21, wherein The metal hydroxide composite is in the form of a plurality of particles.

26. The metal hydroxide complex according to claim 25, wherein The plurality of particles are substantially spherical in shape.

27. The metal hydroxide complex according to claim 25, wherein The plurality of particles have an average particle size of about 10 μm to about 20 μm.

28. The metal hydroxide complex according to claim 21, wherein The metal hydroxide complex exhibits a relative cell viability of at least about 80% or more at a concentration of about 10 μg / ml to about 300 μg / ml.

29. The metal hydroxide complex according to claim 21, wherein The metal hydroxide complex exhibits a relative cell viability of at least about 90% or more at a concentration of about 10 μg / ml to about 300 μg / ml.

30. The metal hydroxide complex according to claim 21, wherein The metal hydroxide complex at a concentration of about 10 μg / ml to about 300 μg / ml increases expression of COL1A1, COL3A1, or a combination thereof.

31. The metal hydroxide complex according to claim 21, wherein The multilayer hydroxide structure is prepared by co-precipitation through a precipitation reaction using alcohol and water as solvents.

32. The metal hydroxide complex according to claim 31, wherein The alcohol is at least one selected from the group consisting of methanol, ethanol, propanol and butanol.

33. The metal hydroxide complex according to claim 21, wherein The metal hydroxide complex is prepared by subjecting an active ingredient and a metal hydroxide structure precursor to co-precipitation through a precipitation reaction using alcohol and water.

34. The metal hydroxide complex according to claim 33, wherein The alcohol is at least one selected from the group consisting of methanol, ethanol, propanol and butanol.

35. The metal hydroxide complex according to claim 33, wherein The ratio of alcohol to water in the co-precipitation is 1:9 to 9:

1.

36. A method for preparing the metal hydroxide complex according to claim 1, comprising subjecting an active ingredient and a metal hydroxide structure precursor to co-precipitation through a precipitation reaction using alcohol and water.

37. The method according to claim 36, wherein The alcohol is at least one selected from the group consisting of methanol, ethanol, propanol and butanol.

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