Amlexanox external preparation for treating psoriasis as well as preparation method and application of amlexanox external preparation

Abstract

The invention provides an amlexanox external preparation for treating psoriasis as well as a preparation method and application of the amlexanox external preparation, and belongs to the technical field of pharmaceutical preparations. The amlexanox external gel is prepared from amlexanox, marine active ingredients, a matrix material, a humectant, a pH regulator and water. The amlexanox gel comprises the following components in percentage by weight: 0.1-5% of amlexanox, 0.1-2% of marine active component, 0.3-3% of matrix material, 0.1-2% of pH regulator, 1-10% of humectant and the balance of water. Test results show that the prepared amlexanox gel is good in in-vitro drug release and transdermal results, uniform in drug content and stable in gel property, and animal experiments show that the amlexanox gel containing marine components can have a good treatment effect on psoriasis.

Description

[0001] This application claims the priority of the Chinese patent application filed with the China Patent Office on January 15, 2024, with application number "202410055632.1" and invention name "A topical preparation of Amlexanox for treating psoriasis, its preparation method and application", the entire contents of which are incorporated by reference in this application. Technical Field

[0002] The present invention belongs to the technical field of pharmaceutical preparations, and in particular relates to an amlexanox external preparation for treating psoriasis, and a preparation method and application thereof. Background Art

[0003] Psoriasis, commonly known as psoriasis, is a chronic and easily recurring inflammatory skin disease with typical clinical manifestations of erythema, rash, plaques, scales, etc. with clear boundaries. At present, the incidence of psoriasis is high and difficult to treat, which seriously affects the life and mental health of patients. Gel preparations have developed rapidly in recent years. They are accepted by more and more people because they are easy to carry, simple to use, and have few adverse reactions. Gel preparations are divided into aqueous gels and oily gels. Carbomer is the most common among aqueous gels. The preparation process of carbomer gel has obvious advantages over traditional creams and ointments, and the product quality is stable. It is especially suitable for the pre-dissolution of water-insoluble drugs in the matrix, improving transdermal efficiency and bioavailability, reducing or avoiding the use of penetration enhancers, thereby reducing the irritation of penetration enhancers to the epidermis of psoriasis patients, improving patient applicability, and playing a positive role in improving the treatment effect.

[0004] Amlexanox, English name: Amlexanox, Chinese chemical name: 2-amino-7-isopropyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-3-carboxylic acid, molecular formula: C 16 H 14 N2O4, molecular weight: 298.29, chemical structure:

[0005]

[0006] Amlexanox is white to pale yellow crystals or crystalline powder, soluble in dimethylformamide, very slightly soluble in methanol or ethanol, and almost insoluble in water. Summary of the invention

[0007] The purpose of the present invention is to provide an amlexanox topical preparation for treating psoriasis, and a preparation method and application thereof. The preparation process of the amlexanox topical preparation is simple and easy to industrialize, wherein the gel has stable performance, good in vitro release, significant effect in treating psoriasis, and is easy to use; and the present invention significantly improves the therapeutic effect on psoriasis after adding brown algae oligosaccharides and fish bladder oligopeptides to the formula.

[0008] In order to achieve the above-mentioned invention object, the present invention is implemented by the following technical solutions:

[0009] The invention provides an amlexanox external preparation for treating psoriasis, wherein the external preparation contains amlexanox.

[0010] Furthermore, the external preparation is an ointment, cream, gel, paste, spray, aerosol, microneedle, patch or lotion.

[0011] Furthermore, the topical preparation is a gel, which contains a matrix material, a moisturizer, a pH adjuster and water; the weight percentage of each component in the amlexanox gel is: 0.1-5% of amlexanox, 0.3-3% of the matrix material, 1-10% of the moisturizer, 0.1-2% of the pH adjuster, and the balance is water.

[0012] Furthermore, the matrix material is one or more of Carbomer 910, Carbomer 934, Carbomer 940, Carbomer 941, and Carbomer 1342; the moisturizer is glycerol; and the pH regulator is triethanolamine or sodium hydroxide.

[0013] Furthermore, the external preparation also contains marine active ingredients, and the weight percentage of the marine active ingredients in the amlexanox gel is 0.1-2%.

[0014] Furthermore, the marine component is one or more of fucoidan, chitosan, brown algae oligosaccharide, chitin, and fish bladder oligopeptide.

[0015] Further, the topical preparation comprises 0.1-5% of amlexanox, 0.3-3% of a matrix material, 1-10% of a moisturizer, 0.1-2% of a pH adjuster, 0.1-2% of a marine active ingredient, and water;

[0016] The marine active ingredients are brown algae oligosaccharides and fish maw oligopeptides; or, the marine active ingredients are brown algae oligosaccharides and fucoidan; or, the marine active ingredients are fucoidan and fish maw oligosaccharides.

[0017] Furthermore, for the scheme in which the marine active ingredients are brown algae oligosaccharides and fish bladder oligopeptides, those skilled in the art can adjust the dosage of brown algae oligosaccharides and fish bladder oligopeptides as needed, for example, the dosage ratio of brown algae oligosaccharides and fish bladder oligopeptides is (1-10): (1:10), optionally, the dosage ratio of brown algae oligosaccharides and fish bladder oligopeptides is (1-5): (1:5), optionally, the dosage ratio of brown algae oligosaccharides and fish bladder oligopeptides is (1-3): (1-3).

[0018] Furthermore, the gel also contains a penetration enhancer and a preservative.

[0019] Furthermore, the penetration enhancer is one or more of laurocapram, peppermint oil, and ethanol; the preservative is one or more of benzalkonium bromide, sodium benzoate, and ethylparaben.

[0020] Furthermore, the weight percentage of the penetration enhancer in the amlexanox gel is 0-1%, and the weight percentage of the preservative in the amlexanox gel is 0.01-1%.

[0021] Furthermore, the particle size and particle size distribution of the amlexanox are D90=15-25 microns, D50=5-9 microns, and D10=1.5-3 microns.

[0022] The present invention also provides a method for preparing the amlexanox gel, comprising the following steps:

[0023] (1) Weigh the prescribed amount of matrix material into sufficient water and allow it to fully swell at room temperature;

[0024] (2) dissolving another marine active ingredient in water and stirring evenly;

[0025] (3) adding a moisturizer to the solution obtained in step (2) and stirring thoroughly;

[0026] (4) After the solution of step (3) is fully mixed with the solution of step (1), a pH adjuster is added dropwise to adjust the pH to 4.5-5, and amlexanox is added in small amounts and multiple times, and the mixture is stirred thoroughly after each addition;

[0027] (5) Add preservatives and continue to stir evenly, add water and stir, add triethanolamine dropwise until pH = 7, add remaining purified water to a sufficient amount, mix evenly, and obtain Amlexanox gel.

[0028] Furthermore, the marine active ingredients are brown algae oligosaccharides and fish bladder oligopeptides, the matrix material is carbomer, and the pH regulator is triethanolamine.

[0029] The present invention also provides the use of the amlexanox external preparation in preparing a medicine for treating psoriasis.

[0030] Compared with the prior art, the advantages and technical effects of the present invention are:

[0031] 1. The pharmaceutical properties of the prepared amlexanox gel were evaluated by in vitro drug release test, transdermal test, content mixing uniformity test and gel stability test. The test results showed that the prepared amlexanox gel had good in vitro drug release and transdermal results, uniform drug content and stable gel properties.

[0032] 2. The present invention further demonstrates through animal experiments that the amlexanox gel containing marine ingredients has a good therapeutic effect on psoriasis, and has a significant improvement effect compared to the use of amlexanox alone.

[0033] 3. The present invention optimizes the marine active ingredients and, by adding brown algae oligosaccharides and fish bladder oligopeptides, can achieve a significant improvement in imiquimod-induced scaling, thickening and erythematous lesions on the back skin of mice, and the combination of brown algae oligosaccharides and fish bladder oligopeptides has a good synergistic effect.

[0034] 4. The present invention provides for the first time an amlexanox gel product for treating psoriasis, and verifies its therapeutic efficacy through a mouse pathological model. However, the prior art has not yet disclosed an amlexanox product that can clearly treat psoriasis, which has important contributions and reference significance for future research and clinical applications of using amlexanox to treat psoriasis. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] The drawings described herein are used to provide a further understanding of the present application and constitute a part of the present application. The illustrative embodiments of the present application and their descriptions are used to explain the present application and do not constitute an improper limitation on the present application. In the drawings:

[0036] Figure 1 These are the IVRT test results of the gels prepared in Examples 1 to 6;

[0037] Figure 2 are the IVRT test results of the gels prepared in Example 2, Example 7 and Example 8;

[0038] Figure 3 is a schematic diagram of the establishment of a psoriasis model and the administration time involved in Example 15;

[0039] Figure 4 This is a photo result of the mouse back skin involved in Example 15;

[0040] Figure 5 This is a picture of the hematoxylin-eosin (H&E) staining result of the back skin on day 8 involved in Example 15;

[0041] Figure 6 is a quantitative result diagram of epidermal thickness involved in Example 15;

[0042] Figure 7 is a graph showing spleen index results of each group on day 8 involved in Example 16;

[0043] Figure 8 This is a graph showing the immunohistochemical results of topical application of Amlexanox gel on inflammatory factors in the skin of psoriasis mice involved in Example 17;

[0044] Fig. 9 The figure is a fluorescence quantitative PCR result diagram of the effect of topical application of Amlexanox gel on the expression of inflammatory factors and epidermal barrier-related proteins in psoriasis mice involved in Example 17;

[0045] Fig.10 This is a Western Blot result diagram of the activation of the MAPK / NF-κB signaling pathway in the skin lesions of psoriasis mice by topical application of amlexanox gel involved in Example 18. DETAILED DESCRIPTION

[0046] The present application is described in detail below in conjunction with examples, but the present application is not limited to these examples. Unless otherwise specified, the raw materials in the examples of the present application are purchased through commercial channels. For example, Amlexanox was purchased from Shandong Keyuan Pharmaceutical Co., Ltd., Carbomer was purchased from Guizhou Xinzihong Pharmaceutical Excipients Co., Ltd., Glycerin was purchased from Hunan Erkang Pharmaceutical Co., Ltd., Ethyl hydroxybenzoate was purchased from Hunan Jiudian Hongyang Pharmaceutical Co., Ltd., Triethanolamine was purchased from Hubei Gedian Renfu Pharmaceutical Excipients Co., Ltd., and Purified water was purchased from Wahaha. Brown algae oligosaccharides were prepared according to the preparation method of brown algae oligosaccharides disclosed in patent CN1161368C, and fish bladder oligopeptides were prepared according to the scheme disclosed in patent CN116640821A or CN117965666A. Specifically, in the present application, fish bladder oligopeptides were prepared according to the scheme disclosed in CN116640821A. Those skilled in the art can also purchase existing commercially available products, such as brown algae oligosaccharides can be purchased from Qingdao Bozhi Huili Biotechnology Co., Ltd., and fish bladder oligopeptides can be purchased from Qingdao Daiyoujia Biotechnology Co., Ltd. It should be noted that those skilled in the art can select and replace the source and manufacturer of the raw materials, and the technical effect of the present application can be achieved.

[0047] The present application scheme is described below through specific embodiments.

[0048] Example 1

[0049] The particle size distribution of amlexanox used in this embodiment is D90=21 μm, D50=8.9 μm, and D10=2.8 μm.

[0050] The formula of the Amlexanox gel in this embodiment is as follows:

[0051]

[0052]

[0053] The preparation method of Amlexanox gel in this embodiment is as follows:

[0054] (1) Weigh 0.75 g of Carbomer 940 and dissolve it in 60 g of water. Let it stand at room temperature to allow the Carbomer to fully swell.

[0055] (2) Dissolve 0.5 g of seaweed oligosaccharide and fish maw oligopeptide in 20 g of water and stir evenly;

[0056] (3) Add 10 g of glycerol to the solution obtained in step (2) and stir thoroughly;

[0057] (4) After the solution of step (3) is fully mixed with the solution of step (1), triethanolamine is added dropwise to adjust the pH to 4.5-5, and the amlexanox is added in small amounts and multiple times, and the mixture is stirred thoroughly after each addition;

[0058] (5) Add preservatives and continue to stir evenly, add 5 g of water and stir, add triethanolamine dropwise until pH = 7, add the remaining purified water to a sufficient amount, mix evenly, and obtain Amlexanox gel.

[0059] IVRT test: vertical Rranz ​​diffusion cell, receiving cell volume 6.5ml, diffusion area 2.4cm 2 The experiment was carried out in phosphate medium with a pH of 6.8, using a 0.45 μm PES disc filter membrane, a temperature of 32 ± 0.5 °C, a stirring speed of 300 rpm, and 2 ml of sample was taken at 0.5, 1, 2, 4, and 6 h, and an equal volume of release solution was replenished. The absorbance of the solution was measured at 350 nm to detect the release amount.

[0060] Example 2

[0061] The particle size distribution of the selected amlexanox is D90=21 μm, D50=8.9 μm, and D10=2.8 μm.

[0062] The formula of the Amlexanox gel in this embodiment is as follows:

[0063]

[0064]

[0065] The preparation method of Amlexanox gel in this embodiment is as follows:

[0066] (1) Weigh the above amounts of Carbomer 910 and Carbomer 940 and add them to 60 g of water, and let stand at room temperature to allow the Carbomer to fully swell;

[0067] (2) Dissolve 0.5 g of seaweed oligosaccharide and fish maw oligopeptide in 20 g of water and stir evenly;

[0068] (3) Add 10 g of glycerol to the solution obtained in step (2) and stir thoroughly;

[0069] (4) After the solution of step (3) is fully mixed with the solution of step (1), triethanolamine is added dropwise to adjust the pH to 4.5-5, and amlexanox is added in small amounts and multiple times, and the mixture is fully stirred after each addition;

[0070] (5) Add preservatives and continue to stir evenly, add 5 g of water and stir, add triethanolamine dropwise until pH = 7, add the remaining purified water to a sufficient amount, mix evenly, and obtain Amlexanox gel.

[0071] The IVRT test was the same as in Example 1.

[0072] Example 3

[0073] The particle size distribution of amlexanox used in this embodiment is D90=21 μm, D50=8.9 μm, and D10=2.8 μm.

[0074] The formula of the Amlexanox gel in this embodiment is as follows:

[0075] Components Dosage Amlexanox 1g Carbomer 934 0.75g Brown algae oligosaccharide 0.5g Fish Maw Oligopeptide 0.5g glycerin 10g Sodium Benzoate 0.1g Sodium hydroxide Moderate Purified water Add to 100g

[0076] The preparation method of Amlexanox gel in this embodiment is as follows:

[0077] (1) Weigh the above amount of Carbomer 934 into 60 g of water and let it stand at room temperature to allow the Carbomer to fully swell;

[0078] (2 Separately, dissolve 0.5 g of seaweed oligosaccharide and fish maw oligopeptide in 20 g of water and stir evenly;

[0079] (3) Add 10 g of glycerol to the solution obtained in step (2) and stir thoroughly;

[0080] (4) After the solution of step (3) is fully mixed with the solution of step (1), sodium hydroxide is added dropwise to adjust the pH to 4.5-5, and amlexanox is added in small amounts and multiple times, and the mixture is stirred thoroughly after each addition;

[0081] (5) Add preservatives and continue to stir evenly, add 5g of water and stir, add sodium hydroxide dropwise until pH = 7, add the remaining purified water until sufficient, and mix evenly to obtain Amlexanox gel.

[0082] The IVRT test was the same as in Example 1.

[0083] Example 4

[0084] The particle size distribution of amlexanox used in this embodiment is D90=15 μm, D50=6.8 μm, and D10=2.0 μm.

[0085] The formula of the Amlexanox gel in this embodiment is as follows:

[0086] Components Dosage Amlexanox 1g Carbomer 941 1.8g Brown algae oligosaccharide 0.5g Fish Maw Oligopeptide 0.5g glycerin 10g Sodium Benzoate 0.1g Sodium hydroxide Moderate Purified water Add to 100g

[0087] The preparation method of Amlexanox gel in this embodiment is as follows:

[0088] (1) Weigh the prescribed amount of Carbomer 941 and dissolve it in 60 g of water. Let it stand at room temperature to allow the Carbomer to fully swell.

[0089] (2 Separately, dissolve 0.5 g of seaweed oligosaccharide and fish maw oligopeptide in 20 g of water and stir evenly;

[0090] (3) Add 10 g of glycerol to the solution obtained in step (2) and stir thoroughly;

[0091] (4) After the solution of step (3) is fully mixed with the solution of step (1), sodium hydroxide is added dropwise to adjust the pH to 4.5-5, and amlexanox is added in small amounts and multiple times, and the mixture is stirred thoroughly after each addition;

[0092] (5) Add preservatives and continue to stir evenly, add 5g of water and stir, add sodium hydroxide dropwise until pH = 7, add the remaining purified water until sufficient, and mix evenly to obtain Amlexanox gel.

[0093] The IVRT test was the same as in Example 1.

[0094] Example 5

[0095] The particle size distribution of amlexanox used in this embodiment is D90=15 μm, D50=6.8 μm, and D10=2.0 μm.

[0096] The formula of the Amlexanox gel in this embodiment is as follows:

[0097]

[0098] The preparation method of Amlexanox gel in this embodiment is as follows:

[0099] (1) Weigh the prescribed amount of Carbomer 940 and Carbomer 1342 into 60 g of water and let stand at room temperature to allow the Carbomer to fully swell;

[0100] (2) Dissolve 0.5 g of seaweed oligosaccharide and fish maw oligopeptide in 20 g of water and stir evenly;

[0101] (3) Add 10 g of glycerol to the solution obtained in step (2) and stir thoroughly;

[0102] (4) After the solution of step (3) is fully mixed with the solution of step (1), triethanolamine is added dropwise to adjust the pH to 4.5-5, and amlexanox is added in small amounts and multiple times, and the mixture is fully stirred after each addition;

[0103] (5) Add preservatives and continue to stir evenly, add 5 g of water and stir, add triethanolamine dropwise until pH = 7, add the remaining purified water until sufficient, and mix evenly to obtain Amlexanox gel.

[0104] The IVRT test was the same as in Example 1.

[0105] Example 6

[0106] The particle size distribution of amlexanox used in this embodiment is D90=81 μm, D50=33 μm, and D10=12 μm.

[0107] The formula of the Amlexanox gel in this embodiment is as follows:

[0108]

[0109]

[0110] The preparation method of Amlexanox gel in this embodiment is as follows:

[0111] (1) Weigh the prescribed amount of Carbomer 940 and dissolve it in 60 g of water. Let it stand at room temperature to allow the Carbomer to fully swell.

[0112] (2) Dissolve 0.5 g of seaweed oligosaccharide and fish maw oligopeptide in 20 g of water and stir evenly;

[0113] (3) Add 10 g of glycerol to the solution obtained in step (2) and stir thoroughly;

[0114] (4) After the solution of step (3) is fully mixed with the solution of step (1), sodium hydroxide is added dropwise to adjust the pH to 4.5-5, and amlexanox is added in small amounts and multiple times, and the mixture is stirred thoroughly after each addition;

[0115] (5) Add preservatives and continue to stir evenly, add 5g of water and stir, add sodium hydroxide dropwise until pH = 7, add the remaining purified water until sufficient, and mix evenly to obtain Amlexanox gel.

[0116] The IVRT test was the same as in Example 1.

[0117] Example 7 Addition of marine active ingredients and penetration enhancers

[0118] The particle size distribution of amlexanox used in this embodiment is D90=21 μm, D50=8.9 μm, and D10=2.8 μm.

[0119] The formula of the Amlexanox gel in this embodiment is as follows:

[0120]

[0121]

[0122] The preparation method of Amlexanox gel in this embodiment is as follows:

[0123] (1) Weigh 0.75 g of Carbomer 940 in 60 g of water and let stand at room temperature to allow the Carbomer to fully swell;

[0124] (2) Dissolve 0.5 g of seaweed oligosaccharide and fish maw oligopeptide in 20 g of water and stir evenly;

[0125] (3) Add 10 g of glycerol to the solution obtained in step (2) and stir thoroughly;

[0126] (4) After the solution of step (3) is fully mixed with the solution of step (1), triethanolamine is added dropwise to adjust the pH to 4.5-5, and amlexanox and laurocapram are added in small amounts and multiple times, and stirred thoroughly after each addition;

[0127] (5) Add preservatives and continue to stir evenly, add 5 g of water and stir, add triethanolamine dropwise until pH = 7, add the remaining purified water until sufficient, and mix evenly to obtain Amlexanox gel.

[0128] The IVRT test was the same as in Example 1.

[0129] Example 8 Addition of penetration enhancer

[0130] The particle size distribution of amlexanox used in this embodiment is D90=21 μm, D50=8.9 μm, and D10=2.8 μm.

[0131] The formula of the Amlexanox gel in this embodiment is as follows:

[0132]

[0133]

[0134] The preparation method of Amlexanox gel in this embodiment is as follows:

[0135] (1) Weigh 0.75 g of Carbomer 940 in 60 g of water and let stand at room temperature to allow the Carbomer to fully swell;

[0136] (2) adding 10 g of glycerol to the solution obtained in step (1) and stirring thoroughly;

[0137] (3) After the solution of step (2) is fully mixed with the solution of step (1), triethanolamine is added dropwise to adjust the pH to 4.5-5, and amlexanox and laurocapram are added in small amounts and multiple times, and stirred thoroughly after each addition;

[0138] (4) Add preservatives and continue to stir evenly, add 5g of water and stir, add triethanolamine dropwise until pH = 7, add the remaining purified water until sufficient, and mix evenly to obtain Amlexanox gel.

[0139] The IVRT test was the same as in Example 1.

[0140] Example 9

[0141] The particle size distribution of amlexanox used in this embodiment is D90=15 μm, D50=6.8 μm, and D10=2.0 μm.

[0142] The formula of the Amlexanox gel in this embodiment is as follows:

[0143] Components Dosage Amlexanox 0.1g Carbomer 941 1.8g Brown algae oligosaccharide 0.5g Fish Maw Oligopeptide 0.5g glycerin 10g Sodium Benzoate 0.1g Sodium hydroxide Moderate Purified water Add to 100g

[0144] The preparation method of Amlexanox gel in this embodiment is as follows:

[0145] (1) Weigh the prescribed amount of Carbomer 941 and dissolve it in 60 g of water. Let it stand at room temperature to allow the Carbomer to fully swell.

[0146] (2) Dissolve 0.5 g of seaweed oligosaccharide and fish maw oligopeptide in 20 g of water and stir evenly;

[0147] (3) Add 10 g of glycerol to the solution obtained in step (2) and stir thoroughly;

[0148] (4) After the solution of step (3) is fully mixed with the solution of step (1), sodium hydroxide is added dropwise to adjust the pH to 4.5-5, and amlexanox is added in small amounts and multiple times, and the mixture is stirred thoroughly after each addition;

[0149] (5) Add preservatives and continue to stir evenly, add 5g of water and stir, add sodium hydroxide dropwise until pH = 7, add the remaining purified water until sufficient, and mix evenly to obtain Amlexanox gel.

[0150] Example 10

[0151] The particle size distribution of amlexanox used in this embodiment is D90=15 μm, D50=6.8 μm, and D10=2.0 μm.

[0152] The formula of the Amlexanox gel in this embodiment is as follows:

[0153] Components Dosage Amlexanox 5g Carbomer 941 1.8g Brown algae oligosaccharide 0.5g Fish Maw Oligopeptide 0.5g glycerin 10g Sodium Benzoate 0.1g Sodium hydroxide Moderate Purified water Add to 100g

[0154] The preparation method of Amlexanox gel in this embodiment is as follows:

[0155] (1) Weigh the prescribed amount of Carbomer 941 and dissolve it in 50 g of water. Let it stand at room temperature to allow the Carbomer to fully swell.

[0156] (2) Dissolve 0.5 g of seaweed oligosaccharide and fish maw oligopeptide in 20 g of water and stir evenly;

[0157] (3) Add 10 g of glycerol to the solution obtained in step (2) and stir thoroughly;

[0158] (4) After the solution of step (3) is fully mixed with the solution of step (1), sodium hydroxide is added dropwise to adjust the pH to 4.5-5, and amlexanox is added in small amounts and multiple times, and the mixture is stirred thoroughly after each addition;

[0159] (5) Add preservatives and continue to stir evenly, add 5g of water and stir, add sodium hydroxide dropwise until pH = 7, add the remaining purified water until sufficient, and mix evenly to obtain Amlexanox gel.

[0160] Example 11 Addition of marine active ingredients and penetration enhancers

[0161] The particle size distribution of amlexanox used in this embodiment is D90=21 μm, D50=8.9 μm, and D10=2.8 μm.

[0162] The formula of the Amlexanox gel in this embodiment is as follows:

[0163] Components Dosage Amlexanox 1g Carbomer 940 0.75g Brown algae oligosaccharide 0.05g Fish Maw Oligopeptide 0.05g glycerin 10g Laurocapram 0.5g Ethylparaben 0.1g Triethanolamine Moderate Purified water Add to 100g

[0164] The preparation method of Amlexanox gel in this embodiment is as follows:

[0165] (1) Weigh 0.75 g of Carbomer 940 in 60 g of water and let stand at room temperature to allow the Carbomer to fully swell;

[0166] (2) Dissolve 0.05 g of seaweed oligosaccharide and fish maw oligopeptide in 20 g of water and stir evenly;

[0167] (3) Add 10 g of glycerol to the solution obtained in step (2) and stir thoroughly;

[0168] (4) After the solution of step (3) is fully mixed with the solution of step (1), triethanolamine is added dropwise to adjust the pH to 4.5-5, and amlexanox and laurocapram are added in small amounts and multiple times, and stirred thoroughly after each addition;

[0169] (5) Add preservatives and continue to stir evenly, add 5 g of water and stir, add triethanolamine dropwise until pH = 7, add the remaining purified water until sufficient, and mix evenly to obtain Amlexanox gel.

[0170] Example 12 Addition of marine active ingredients and penetration enhancers

[0171] The particle size distribution of amlexanox used in this embodiment is D90=21 μm, D50=8.9 μm, and D10=2.8 μm.

[0172] The formula of the Amlexanox gel in this embodiment is as follows:

[0173] Components Dosage Amlexanox 1g Carbomer 940 0.75g Brown algae oligosaccharide 1g Fish Maw Oligopeptide 1g glycerin 10g Laurocapram 0.5g Ethylparaben 0.1g Triethanolamine Moderate Purified water Add to 100g

[0174] The preparation method of Amlexanox gel in this embodiment is as follows:

[0175] (1) Weigh 0.75 g of Carbomer 940 in 60 g of water and let stand at room temperature to allow the Carbomer to fully swell;

[0176] (2) Dissolve 1 g of algae oligosaccharide and 1 g of fish maw oligopeptide in 20 g of water and stir evenly;

[0177] (3) Add 10 g of glycerol to the solution obtained in step (2) and stir thoroughly;

[0178] (4) After the solution of step (3) is fully mixed with the solution of step (1), triethanolamine is added dropwise to adjust the pH to 4.5-5, and amlexanox and laurocapram are added in small amounts and multiple times, and stirred thoroughly after each addition;

[0179] (5) Add preservatives and continue to stir evenly, add 5 g of water and stir, add triethanolamine dropwise until pH = 7, add the remaining purified water until sufficient, and mix evenly to obtain Amlexanox gel.

[0180] Example 13 Addition of marine active ingredients and penetration enhancers

[0181] The particle size distribution of amlexanox used in this embodiment is D90=21 μm, D50=8.9 μm, and D10=2.8 μm.

[0182] The formula of the Amlexanox gel in this embodiment is as follows:

[0183] Components Dosage Amlexanox 1g Carbomer 940 0.75g Brown algae oligosaccharide 0.3g Fish Maw Oligopeptide 0.3g glycerin 10g Laurocapram 0.5g Ethylparaben 0.1g Triethanolamine Moderate Purified water Add to 100g

[0184] The preparation method of Amlexanox gel in this embodiment is as follows:

[0185] (1) Weigh 0.75 g of Carbomer 940 in 60 g of water and let stand at room temperature to allow the Carbomer to fully swell;

[0186] (2) Dissolve 0.3 g of algae oligosaccharide and fish maw oligopeptide in 20 g of water and stir evenly;

[0187] (3) Add 10 g of glycerol to the solution obtained in step (2) and stir thoroughly;

[0188] (4) After the solution of step (3) is fully mixed with the solution of step (1), triethanolamine is added dropwise to adjust the pH to 4.5-5, and amlexanox and laurocapram are added in small amounts and multiple times, and stirred thoroughly after each addition;

[0189] (5) Add preservatives and continue to stir evenly, add 5 g of water and stir, add triethanolamine dropwise until pH = 7, add the remaining purified water until sufficient, and mix evenly to obtain Amlexanox gel.

[0190] Embodiment 14

[0191] 1g of each gel prepared in Examples 1 to 13 was placed in a 1.5ml centrifuge tube, centrifuged at 4500rpm for 15min, the appearance was observed, and 3 samples were taken from the upper and lower layers of the centrifuge tube, and the RSD of 6 samples in each centrifuge tube was calculated to detect the uniformity of content. The results show that the particle size and distribution of the raw materials have a significant effect on the stability of the gel. When the particle size and distribution of the raw materials are similar, the content of the gels in different examples is stable. The results are shown in Table 1.

[0192] Table 1 Stability and content results of gelling agents in Examples 1 to 13

[0193]

[0194]

[0195] As shown in Table 1, except for Example 6, the gels prepared in the other examples have good appearance and uniform and stable content distribution. It can be seen from the results of Example 6 that the particle size of the raw material becomes larger, which will affect the content uniformity of the preparation under extreme conditions. Therefore, the present invention experimentally determines that the particle size and particle size distribution of amlexanox are D90=15-25 microns, D50=5-8 microns, and D10=1.5-3 microns.

[0196] like Figure 1 and Figure 2 As shown, the in vitro release rate R of the gel prepared in the embodiment of the present invention is 2 All are greater than 0.9. Figure 2 As shown, through Examples 2, 7 and 8, it is shown that the addition of marine active ingredients and penetration enhancers has no significant effect on IVRT, but the animal efficacy data of topical treatment of psoriasis proves that the addition of marine active ingredients and penetration enhancers significantly improves the therapeutic effect on psoriasis.

[0197] Example 15 Construction of Imiquimod-induced Psoriasis-like Skin Lesion Model in Mice and Evaluation of Drug Activity

[0198] Schematic diagram of psoriasis model establishment and drug administration time Figure 3As shown. Male BALB / c mice were randomly divided into 6 groups according to body weight, including a normal group (Normal), a model group (imiquimod (IMQ), Model), a positive control (dexamethasone, DXMS), and a drug-treated group (amlexanox gel, AML; amlexanox + fish bladder oligopeptide + brown algae oligosaccharide gel, AML+; amlexanox + fish bladder oligopeptide + brown algae oligosaccharide + penetration-enhancing gel, AML++), with 8 mice in each group, wherein AML++ in the drug-treated group was the scheme of Example 7, while AML+ lacked the penetration-enhancing gel compared to the scheme of Example 7, and AML lacked fish bladder oligopeptide, brown algae oligosaccharide and penetration-enhancing gel compared to Example 7. Skin preparation was performed 2 days before the start of modeling. Except for the mice in the normal group, 62.5 mg of 5% imiquimod was applied to the prepared skin area every morning for 7 consecutive days to induce psoriasis-like skin lesions. Two days after modeling, the drug group was given drug treatment (62.5 mg / mouse) at the prepared skin site every afternoon for 5 consecutive days.

[0199] The degree of skin damage on the back of the mice was observed from three aspects: scaling, skin thickening, and erythema. The therapeutic effect of the drug was recorded by taking photos. The scoring criteria were as follows: 0 points, no symptoms; 1 point; mild symptoms; 2 points, moderate symptoms; 3 points, moderate to severe symptoms; 4 points, severe symptoms. The total PASI score was calculated to characterize the incidence of psoriasis-like skin damage and the therapeutic effect of the drug. On the 8th day of the experiment, the mice were euthanized, the skin was collected, fixed with paraformaldehyde, and used for subsequent HE staining, taking photos, and calculating the epidermal thickness. The results are as follows: Figure 5 and Figure 6 As shown. Figure 5 This is the result of hematoxylin-eosin (H&E) staining of the back skin on day 8. Figure 6 The quantitative results of epidermal thickness are expressed as mean ± SD (n = 8). *p < 0.05, **p < 0.01, ***p < 0.001, compared with the normal group; #p < 0.05, ##p < 0.01, ###p < 0.001, compared with the model group.

[0200] The results of scaling, skin thickening, erythema, and PASI total score are shown in Tables 2 to 6 below.

[0201] Table 2 Scale test results

[0202]

[0203] Table 3 Thickening test results

[0204]

[0205]

[0206] Table 4 Erythema test results

[0207]

[0208] Table 5 PASI total score test results

[0209]

[0210] Table 6 Calculation results of mean values ​​of scaling, thickening, erythema and PASI total score

[0211]

[0212] according to Figure 4 As shown in Tables 2 to 6, after 7 days of applying imiquimod cream to the back skin of mice, it can be seen that obvious scaling, thickening and erythematous lesions appeared on the back skin of mice, and the epidermal thickness was significantly thickened compared with the normal group. Topical dexamethasone and the amlexanox gel of the present invention can significantly improve these pathological symptoms, and the epidermal thickness is restored. After adding fish bladder oligopeptides and brown algae oligosaccharide gel, the effect is better than that of amlexanox gel alone, and the effect is best after adding a penetration-enhancing gel. The above results show that topical amlexanox gel protects psoriasis-like skin lesions induced by imiquimod in mice.

[0213] according to Figure 5 and Figure 6 It can be seen from the pathology that after applying imiquimod cream to the back skin of mice for 7 days, the thickness of the mouse epidermis was significantly thickened compared with the normal group. External use of dexamethasone and the amlexanox gel of the present invention can significantly reduce the thickness of the epidermis, and after adding fish bladder oligopeptides and brown algae oligosaccharide gel, the effect is better than that of the amlexanox gel alone. The effect is best after adding the penetration-enhancing gel, and the epidermal thickness is close to that of the normal group. It shows that external use of amlexanox gel improves the psoriasis-like skin thickening phenomenon induced by imiquimod in mice.

[0214] Example 16 Effect of Amlexanox on Spleen Function in Imiquimod-Induced Psoriasis Mice

[0215] At the end of the experiment in Example 15, the spleen of the mice was weighed, and the spleen index was calculated based on the weight of the mice.

[0216] Figure 7 The effect of topical application of Amlexanox cream and its preparations on the spleen index (spleen weight / body weight) of psoriasis mice. Data are expressed as mean±SD (n=8), *p<0.05, **p<0.01, ***p<0.001, compared with the normal group; #p<0.05, ##p<0.01, ###p<0.001, compared with the model group.

[0217] like Figure 7As shown in the results, the use of imiquimod can significantly increase the spleen index of mice and increase the inflammatory response of the spleen, while the external use of amlexanox gel can significantly reduce the spleen index. After adding fish bladder oligopeptides and brown algae oligosaccharide gel, the effect is better than that of amlexanox gel alone, and the effect is best after adding the penetration-enhancing gel. This shows that amlexanox can regulate the inflammatory response caused by psoriasis and play a certain anti-inflammatory role.

[0218] Example 17 Effect of Amlexanox on the Expression of Skin Inflammatory Factors and Epidermal Barrier-Related Proteins in Imiquimod-Induced Psoriasis Mice

[0219] At the end of the experiment in Example 15, a small piece of mouse skin was taken and fixed, and the expression of inflammatory factors TNF-α, IL-17A, IL-6, IL-1βand IL-22 in the skin was detected by immunohistochemistry; a small piece of skin was taken, RNA was extracted, reverse transcribed, and qPCR was used to detect the expression of inflammatory factors TNFα, IL-17A, IL-17F, IL-22, IL-23, IL-1β, and IL-6; a small piece of skin was taken, RNA was extracted, reverse transcribed, and qPCR was used to detect the expression of epidermal barrier-related proteins aquaporin 3 (AQP3) and filaggrin (FLG).

[0220] Figure 8 The effect of topical application of Amlexanox gel on inflammatory factors in the skin of psoriasis mice. The figure shows the results of immunohistochemical staining of TNF-α, IL-17A, IL-6, IL-22 and IL-1β in the back skin on the 8th day, magnified 200 times. Fig. 9 The effect of topical application of amlexanox gel on the expression of related inflammatory factors and epidermal barrier-related proteins in psoriasis mice, where A is the level of TNF-α, IL-17A, IL-17F, IL-6, IL-22, IL-1β and IL-23 in skin tissue detected by Real-time PCR, and B is the level of AQP3 and FLG in skin detected by Real-time PCR. Data are expressed as mean±SD (n=8), *p<0.05, **p<0.01, ***p<0.001, compared with the normal group; #p<0.05, ##p<0.01, ###p<0.001, compared with the model group.

[0221] according to Figure 8 , Fig. 9 The results showed that immunohistochemistry results showed that the use of imiquimod could upregulate the expression of inflammatory factors in the mouse skin and increase the inflammatory response, while the topical application of amlexanox gel could significantly reduce the expression of inflammatory factors. qPCR results showed that the transcription levels of TNFα, IL-17A, IL-17F, IL-22, IL-23, IL-1β, IL-6 and other genes induced by imiquimod were significantly increased, while topical amlexanox gel could significantly reduce the expression of each factor; the expression of AQP3 and FLG induced by imiquimod was significantly reduced, while topical amlexanox gel could significantly increase the expression of AQP3 and FLG. Compared with the single amlexanox gel, the addition of fish bladder oligopeptides and brown algae oligosaccharides significantly reduced the inflammatory factors, and the effect was best after the addition of penetration enhancers. In summary, amlexanox can regulate the inflammatory response of psoriatic skin and exert anti-inflammatory effects; regulate the skin barrier dysfunction caused by psoriasis and enhance the barrier function of the skin.

[0222] Example 18 Effect of Amlexanox on the Activation of MAPK / NF-κB Signaling Pathway in Imiquimod-Induced Psoriasis Mouse Lesions

[0223] At the end of the experiment in Example 15, a small piece of mouse skin was taken to extract protein, and the protein concentration was detected by BCA kit. The expression levels of p-p38 MAPK / p38 MAPK and p-NF-κB p65 / NF-κB p65 proteins were detected by Western blot, and grayscale analysis was performed using Image J.

[0224] Fig.10 The effect of topical application of amlexanox gel on the activation of MAPK / NF-κB signaling pathway in psoriasis mouse lesions. A: Western blot analysis of the back skin of mice. GAPDH was used as an internal reference. B-C: p-p38MAPK / p38 MAPK and p-NF-κB p65 / NF-κB p65 were quantitatively analyzed by Image J. Data are expressed as mean ± SD (n = 3), *p < 0.05, **p < 0.01, ***p < 0.001, compared with the normal group; #p < 0.05, ##p < 0.01, ###p < 0.001, compared with the model group.

[0225] according to Fig.10 The results showed that the use of imiquimod significantly enhanced the expression levels of p-p38 MAPK / p38 MAPK and p-NF-κB p65 / NF-κB p65 proteins, and topical amlexanox gel significantly reduced the expression levels of inflammation-related proteins. The above results indicate that topical amlexanox gel can inhibit the activation of the MAPK / NF-κB inflammatory signaling pathway.

[0226] Example 19 Exploration of therapeutic effects of marine active ingredients

[0227] The experimenters explored the different therapeutic effects of different combinations of marine active ingredients on psoriasis, wherein the sample components are basically the same as those in Example 7, except that the types of marine active ingredients are different. For specific differences, see Table 7 below.

[0228] Table 7 Types and dosages of marine active ingredients

[0229] Sample No. Types of marine active ingredients and their dosage Example 7 Brown algae oligosaccharide 0.5g, fish maw oligopeptide 0.5g Embodiment 11 Brown algae oligosaccharide 0.05g, fish maw oligopeptide 0.05g Example 12 Brown algae oligosaccharide 1g, fish maw oligopeptide 1g Embodiment 13 Brown algae oligosaccharide 0.3g, fish maw oligopeptide 0.3g Experimental sample 1# Brown algae oligosaccharide 0.5g, chitosan 0.5g Experimental sample 2# Brown algae oligosaccharide 0.5g, Fucoidan 0.5g Experimental sample 3# Brown algae oligosaccharide 0.5g, chitin 0.5g Experimental sample 4# Chitosan 0.5g, fish bladder oligopeptide 0.5g Experimental sample 5# Fucoidan 0.5g, fish maw oligopeptide 0.5g Experimental sample 6# Chitin 0.5g, fish bladder oligopeptide 0.5g Experimental sample 7# Chitosan 0.5g, chitin 0.5g Experimental sample 8# Fish Maw Oligopeptide 1.0g Experimental sample 9# Brown algae oligosaccharide 1.0g

[0230] The same detection method as before was used to detect the therapeutic effects of experimental samples 1#~9# on psoriasis mice. The calculation results of the mean values ​​of scaling, thickening, erythema and PASI total score are shown in Table 8 below.

[0231] Table 8 Calculation results of mean values ​​of scaling, thickening, erythema and PASI total score of test samples

[0232] serial number Scaly Thickening erythema PASI total score Example 7 0.25 0.25 0 0.5 Embodiment 11 0.75 0.625 0.5 1.875 Example 12 0.25 0.375 0 0.625 Embodiment 13 0.32 0.39 0.125 0.835 Experimental sample 1# 0.625 0.5 0.375 1.5 Experimental sample 2# 0.625 0.5 0.25 1.375 Experimental sample 3# 0.625 0.5 0.375 1.5 Experimental sample 4# 0.75 0.625 0.5 1.875 Experimental sample 5# 0.75 0.625 0.375 1.75 Experimental sample 6# 0.75 0.625 0.5 1.875 Experimental sample 7# 0.875 0.75 0.5 2.125 Experimental sample 8# 0.5 0.375 0.125 1 Experimental sample 9# 0.375 0.375 0.125 0.875

[0233] According to the results in the above table, among the selection of marine active ingredients, the solution of adding brown algae oligosaccharides and fish bladder oligopeptides has the best effect on improving the scaling, thickening and erythematous lesions on the back skin of mice induced by imiquimod, and according to the results of experimental samples 8# and 9#, it can be seen that the combination of brown algae oligosaccharides and fish bladder oligopeptides has a good synergistic effect.

[0234] The above is only the embodiment of the present application, and the protection scope of the present application is not limited by these specific embodiments, but is determined by the claims of the present application. For those skilled in the art, the present application can have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the technical ideas and principles of the present application should be included in the protection scope of the present application.

Claims

1. An amlexanox topical preparation for treating psoriasis, characterized in that The external preparation contains amlexanox.

2. The amlexanox external preparation according to claim 1, characterized in that The external preparation is an ointment, a cream, a gel, a paste, a spray, an aerosol, a microneedle, a medicated patch or a lotion.

3. The amlexanox external preparation according to claim 2, characterized in that: The external preparation is a gel, which contains a matrix material, a moisturizer, a pH regulator and water; the weight percentage of each component in the amlexanox gel is: 0.1-5% of amlexanox, 0.3-3% of the matrix material, 1-10% of the moisturizer, 0.1-2% of the pH regulator, and the balance is water.

4. The amlexanox external preparation according to claim 3, characterized in that: The matrix material is one or more of Carbomer 910, Carbomer 934, Carbomer 940, Carbomer 941 and Carbomer 1342; the moisturizing agent is glycerol; and the pH regulator is triethanolamine or sodium hydroxide.

5. The amlexanox external preparation according to claim 3, characterized in that: The gel also contains marine active ingredients, and the weight percentage of the marine active ingredients in the amlexanox gel is 0.1-2%.

6. The amlexanox external preparation according to claim 5, characterized in that: The marine component is one or more of fucoidan, chitosan, brown algae oligosaccharide, chitin and fish bladder oligopeptide.

7. The amlexanox external preparation according to claim 6, characterized in that: The topical preparation comprises 0.1-5% of amlexanox, 0.3-3% of a matrix material, 1-10% of a moisturizer, 0.1-2% of a pH regulator, 0.1-2% of a marine active ingredient and water; The marine active ingredients are brown algae oligosaccharides and fish maw oligopeptides; or, the marine active ingredients are brown algae oligosaccharides and fucoidan; or, the marine active ingredients are fucoidan and fish maw oligosaccharides.

8. The amlexanox external preparation according to claim 1, characterized in that: The particle size and particle size distribution of the amlexanox are D90=15-25 microns, D50=5-9 microns, and D10=1.5-3 microns.

9. The method for preparing the amlexanox gel according to claim 5, characterized in that The steps include: (1) Weigh the prescribed amount of matrix material into sufficient water and allow it to fully swell at room temperature; (2) dissolving another marine active ingredient in water and stirring evenly; (3) adding a moisturizer to the solution obtained in step (2) and stirring thoroughly; (4) After the solution of step (3) is fully mixed with the solution of step (1), a pH adjuster is added dropwise to adjust the pH to 4.5-5, and amlexanox is added in small amounts and multiple times, and the mixture is stirred thoroughly after each addition; (5) Add preservatives and continue to stir evenly, add water and stir, add triethanolamine dropwise until pH = 7, add remaining purified water to a sufficient amount, mix evenly, and obtain Amlexanox gel.

10. Use of the amlexanox external preparation according to any one of claims 1 to 8 in the preparation of a medicament for treating psoriasis.

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