Specific drug for treating psoriasis, compound and preparation method thereof

Abstract

The invention belongs to the field of medicines, and relates to a special-effect medicine for treating psoriasis, a compound and a preparation method thereof. The invention provides an application of Neo-A and a composition thereof in treating autoimmune related skin diseases, and a preparation method of the composition, and particularly provides a new drug for treating psoriasis, which is remarkable in curative effect.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, relates to new uses of known compounds, and particularly relates to the use of neo-A of Salvia przewalskii Maxim. and its composition in the preparation of drugs for treating autoimmune-related skin diseases. Background Art

[0002] Psoriasis is an autoimmune-mediated inflammatory skin disease induced by both internal genetics and external environment, and is mainly clinically characterized by erythema, papules, and scale lesions. According to relevant statistical data, about 120 million people worldwide suffer from psoriasis, accounting for 2% of the global population. The long-term disease management has brought serious troubles to the lives of patients, and it is also a key research disease worldwide. The specific pathogenesis of psoriasis is not yet clear, and it still cannot be completely cured at present. The increase in pro-inflammatory cytokines released by immune cells and the chronic activation of the innate immune system and the acquired immune system are the main reasons for the long-term damage of various tissues and organs. At present, the commonly used drugs for the clinical treatment of psoriasis mainly include immunosuppressants, retinoids, cyclosporine, tacrolimus, glucocorticoids, antibiotics, etc. These drugs can relieve symptoms to a certain extent, but are prone to drug resistance and adverse reactions. Therefore, it is very necessary to develop new drugs for the treatment of psoriasis with significant efficacy and high safety.

[0003] Neo-A of Salvia przewalskii Maxim. is a phenanthraquinone derivative isolated from the Chinese herbal medicine Salvia miltiorrhiza Bunge, and its molecular formula is C 36 H 28 O 6 as shown in formula (1). Modern pharmacological studies have shown that it has obvious immunomodulatory functions.

[0004] Summary of the Invention

[0005] The new use of Neo-A of the present invention relates to the use of Neo-A and its composition in the preparation of drugs for treating autoimmune-related skin diseases, especially in the preparation of drugs for treating psoriasis.

[0006] On the one hand, the present invention provides the use of neo-A of Salvia przewalskii Maxim. in the preparation of drugs or medicine kits for treating autoimmune-related skin diseases.

[0007] In some embodiments, the autoimmune-related skin diseases include psoriasis, rheumatoid arthritis, systemic lupus erythematosus, atopic dermatitis, vitiligo, pemphigus or dermatomyositis; preferably psoriasis.

[0008] In some embodiments, the drug or medicine kit further comprises tacrolimus (TACR).

[0009] In some embodiments, the drug or the medicine box contains a pharmaceutical composition with Neo-A and / or TACR as the active ingredient.

[0010] In some embodiments, the Neo-A can be used in combination with tacrolimus.

[0011] In some embodiments, the pharmaceutical composition further contains pharmaceutically acceptable auxiliary components, such as humectants, lubricants, solubilizers, emulsifiers, antibacterial agents, solvents, etc.

[0012] In some embodiments, the dosage forms of the pharmaceutical composition include one or more of topical application dosage forms and solution dosage forms.

[0013] In some embodiments, the dosage form of the pharmaceutical composition is a topical application dosage form.

[0014] In some embodiments, the dosage form of the pharmaceutical composition is a gel, cream or ointment.

[0015] In some embodiments, the pharmaceutical composition is a topical application dosage form, and the auxiliary components are selected from one or more of humectants, lubricants, solubilizers, emulsifiers, antibacterial agents, solvents.

[0016] In some embodiments, the humectant is petrolatum, glycerol and / or hexylene glycol.

[0017] In some embodiments, three different humectants are used, wherein the first humectant is glycerol, the second humectant is petrolatum, and the third humectant is hexylene glycol.

[0018] In some embodiments, the lubricant is dimethicone.

[0019] In some embodiments, the solubilizer is PEG.

[0020] In some embodiments, the emulsifier is cetyl alcohol.

[0021] In some embodiments, the antibacterial agent is p-hydroxyacetophenone.

[0022] In some embodiments, the solvent is water.

[0023] In some embodiments, the auxiliary components are as follows: the first humectant is glycerol, the second humectant is petrolatum, the third humectant is hexylene glycol, the lubricant is dimethicone, the solubilizer is PEG, the emulsifier is cetyl alcohol, the antibacterial agent is p-hydroxyacetophenone, and the solvent is water.

[0024] In some embodiments, calculated based on the total weight of the pharmaceutical composition, the weight percentage of the humectant is 10.0 - 20.0%, such as 10.0%, 10.5%, 11.0%, 11.5%, 12.0%, 12.5%, 13.0%, 13.5%, 14.0%, 14.5%, 15.0%, 15.5%, 16.0%, 16.5%, 17.0%, 17.5%, 18.0%, 18.5%, 19.0%, 19.5% or 20.0%, preferably 15.0 - 17.0%, more preferably 16.5%.

[0025] In some embodiments, calculated based on the total weight of the pharmaceutical composition, the weight percentage of the lubricant is 1.0 - 10.0%, such as 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 7.5%, 8.0%, 8.5%, 9.0%, 9.5% or 10.0%, preferably 2.0 - 6.0%, more preferably 4.0%.

[0026] In some embodiments, calculated based on the total weight of the pharmaceutical composition, the weight percentage of the solubilizer is 1.0 - 5.0%, such as 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5% or 5.0%, preferably 1.0 - 3.0%, more preferably 2.0%.

[0027] In some embodiments, calculated based on the total weight of the pharmaceutical composition, the weight percentage of the emulsifier is 1.0 - 5.0%, such as 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5% or 5.0%, preferably 1.0 - 3.0%, more preferably 1.5%.

[0028] In some embodiments, calculated based on the total weight of the pharmaceutical composition, the weight percentage of the antibacterial agent is 0.05 - 1.0%, such as 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95% or 1.0%, preferably 0.1 - 0.5%, more preferably 0.3%.

[0029] In some embodiments, in the pharmaceutical composition, calculated based on the total weight of the composition, the ratio of each component is: humectant 16.5%, lubricant 4%, solubilizer 2%, emulsifier 1.5%, antibacterial agent 0.3%.

[0030] In some embodiments, the content of Neo-A in the pharmaceutical composition is 0.2 - 8.0 μM, and can be, for example, 0.2, 0.3, 0.4, 0.5, 0.54, 0.6, 0.7, 0.8, 0.9, 1.0, 1.8, 2.0, 3.0, 4.0, 5.0, 5.4, 6.0, 7.0, or 8.0 μM.

[0031] In some embodiments, the content of Neo-A is 0.54 - 5.4 μM.

[0032] In some embodiments, the content of Neo-A is 0.54 - 1.0 μM, or 1.0 - 3.0 μM, or 4.0 - 5.4 μM.

[0033] In some embodiments, the content of Neo-A is 0.54 μM.

[0034] In some embodiments, the content of Neo-A is 1.8 μM.

[0035] In some embodiments, the content of Neo-A is 5.4 μM.

[0036] In some embodiments, the product dose of the pharmaceutical composition is 160 - 200 mg, and can be, for example, 160, 165, 170, 175, 180, 185, 190, 195, 200 mg, preferably 180 mg.

[0037] In some embodiments, the composition is prepared as follows:

[0038] (1) Mix the solvent, the first humectant, and the solubilizer and heat.

[0039] (2) Add the second humectant and the emulsifier in sequence.

[0040] (3) Keep the heating state and stir.

[0041] (4) Stir and cool down, and add the antibacterial agent.

[0042] (5) Stir the lubricant and the therapeutic drug evenly and add them after cooling.

[0043] (6) Add the third humectant after cooling.

[0044] In some embodiments, the heating in step (1) and / or step (3) is heating to 75 - 85 °C, and can be, for example, 75 °C, 76 °C, 77 °C, 78 °C, 79 °C, 80 °C, 81 °C, 82 °C, 83 °C, 84 °C, 85 °C, preferably 80 °C.

[0045] In some embodiments, the stirring in step (3) is carried out for 20 - 40 min, for example, it can be 20, 25, 30, 35, or 40 min, preferably 30 min.

[0046] In some embodiments, the temperature reduction in step (4) is to reduce the temperature to 45 - 55 °C, for example, it can be 45 °C, 46 °C, 47 °C, 48 °C, 49 °C, 50 °C, 51 °C, 52 °C, 53 °C, 54 °C, 55 °C, preferably 50 °C.

[0047] In some embodiments, the stirring in step (5) is carried out at room temperature.

[0048] In some embodiments, the temperature reduction in step (5) is to reduce the temperature to 35 - 45 °C, for example, it can be 35 °C, 36 °C, 37 °C, 38 °C, 39 °C, 40 °C, 41 °C, 42 °C, 43 °C, 44 °C, 45 °C, preferably 40 °C.

[0049] In some embodiments, the temperature reduction in step (6) is to reduce the temperature to room temperature.

[0050] In some embodiments, the room temperature is 20 - 25 °C, for example, it can be 20 °C, 21 °C, 22 °C, 23 °C, 24 °C, 25 °C.

[0051] In some embodiments, the first humectant is glycerol.

[0052] In some embodiments, the second humectant is petrolatum.

[0053] In some embodiments, the third humectant is hexylene glycol.

[0054] In some embodiments, the lubricant is dimethyl silicone oil.

[0055] In some embodiments, the solubilizer is polyethylene glycol (PEG).

[0056] In some embodiments, the emulsifier is cetyl alcohol.

[0057] In some embodiments, the antibacterial agent is p - hydroxyacetophenone.

[0058] In some embodiments, the solvent is water.

[0059] In some embodiments, the therapeutic drug is Neo - A.

[0060] In some embodiments, the first humectant is glycerol, the second humectant is petrolatum, the third humectant is hexylene glycol, the lubricant is dimethyl silicone oil, the solubilizer is PEG, the emulsifier is cetyl alcohol, the antibacterial agent is p - hydroxyacetophenone, and the solvent is water.

[0061] In some embodiments, the therapeutic drug is Neo-A combined with TCAR.

[0062] In some embodiments, the composition is prepared as follows:

[0063] (1) Mix water, PEG and glycerol and heat to 80 °C;

[0064] (2) Add petrolatum and cetyl alcohol in sequence;

[0065] (3) Stir at 80 °C for 30 min;

[0066] (4) Stir and cool down to 50 °C and add p-hydroxyacetophenone;

[0067] (5) Stir dimethicone and Neo-A evenly at room temperature and add them when cooled down to 40 °C;

[0068] (6) Cool down to room temperature and add hexylene glycol.

[0069] On the other hand, the present invention provides a drug or medicine kit for treating autoimmune-related skin diseases, which comprises Neo-A and / or TACR.

[0070] In some embodiments, the autoimmune-related skin diseases include psoriasis, rheumatoid arthritis, systemic lupus erythematosus, atopic dermatitis, vitiligo, pemphigus or dermatomyositis; preferably psoriasis.

[0071] On the other hand, the present invention provides a method for treating autoimmune-related skin diseases, the steps of which include administering the above-mentioned pharmaceutical composition to a subject.

[0072] In some embodiments, the autoimmune-related skin diseases include psoriasis, rheumatoid arthritis, systemic lupus erythematosus, atopic dermatitis, vitiligo, pemphigus or dermatomyositis; preferably psoriasis.

[0073] Advantages of the present invention

[0074] (1) Provide the use of Neo-A in the treatment of autoimmune-related skin diseases, including its composition and preparation method;

[0075] (2) The Neo-A cream of the present invention has a good therapeutic effect on IMIQ-induced psoriasis mice at different administration doses, and the effect is equivalent to that of the positive drug TACR. The difference is that Neo-A has no effect on the spleen index;

[0076] (3) The therapeutic effect of the high-dose Neo-A cream combined with TACR of the present invention on IMIQ-induced psoriasis mice is better than that of Neo-A cream and TACR administered alone;

[0077] (4) The cream formulation of the medicament for treating skin diseases of the present invention is more easily absorbed compared with conventional spraying administration. Description of the Drawings

[0078] Figure 1 The experimental flow chart showing the therapeutic effects of Neo-A and its formulation (Neo-A and TACR) on IMIQ (imiquimod)-induced psoriasis model mice.

[0079] Figure 2 Showing the effects of Neo-A and its formulation (Neo-A and TACR) on the dorsal skin morphology of IMIQ-induced psoriasis model mice in Example 3. Among them, Figure A is a photograph of typical lesioned mice in each group on the 7th day of drug treatment; Figure B is the change in the severity score of skin lesions in each group of mice every day after drug treatment; Figure C is the severity score of skin lesions in each group of mice after drug treatment on the 7th day (N = 7-14, mean ± SEM, **p < 0.01, ***p < 0.001, ****p < 0.0001).

[0080] Figure 3 Showing the effects of Neo-A and its formulation (Neo-A and TACR) on the dorsal skin histopathology of IMIQ-induced psoriasis model mice in Example 4. Among them, Figure A is the histological morphology of the dorsal skin of typical mice in each group on the 7th day of drug treatment; Figure B is the Baker score of the dorsal skin of each group of mice on the 7th day of drug treatment; Figure C is the dorsal skin thickness of each group of mice on the 7th day of drug treatment (N = 7-14, mean ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).

[0081] Figure 4 Showing the effects of Neo-A and its formulation (Neo-A and TACR) on the body weight (A) and spleen index (B) of IMIQ-induced psoriasis model mice in Example 5 (N = 7-14, mean ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001). Detailed Description of the Invention

[0082] For the purposes of clear and concise description, features are described herein as part of the same or separate embodiments. However, it will be understood that the scope of the present disclosure may include embodiments having combinations of all or some of the described features. The technical solutions of the present invention will be described clearly and completely below. Obviously, all other embodiments obtained by those of ordinary skill in the art based on the specific embodiments of the present invention without creative efforts shall fall within the scope of protection of the present invention.

[0083] I. Definitions

[0084] Unless otherwise specified, the following terms used in the specification and claims have the following meanings:

[0085] The numerical ranges used in the present disclosure should be understood to have enumerated all the numbers within that range. For example, the range of 1 to 20 should be understood to include any number, combination of numbers, or sub-range from the following group: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.

[0086] As shown in the present disclosure, the term "comprises" or "comprising" means "including but not limited to". This term is intended to be open-ended, specifying the presence of any of the stated features, elements, integers, steps, or components, but not excluding the presence or addition of one or more other features, elements, integers, steps, components, or groups thereof. Thus, the term "comprising" includes the more restrictive terms "consisting of" and "consisting essentially of". In one embodiment, the term "comprising" used throughout the application, particularly in the claims, may be replaced by the term "consisting of".

[0087] As shown in the present disclosure, the terms "optionally", "any one", "any", or "any item" mean that the subsequently described event or circumstance may or may not occur, and this description includes the cases where the event or circumstance occurs or does not occur. As used in the present invention, "a" and "an" are used in the present invention to refer to one or more than one grammatical object.

[0088] As shown in the present disclosure, "and / or" should be understood to mean any one of the options or a combination of any two or more of the options.

[0089] As used herein, the term "auxiliary ingredient" refers to an important ingredient that has important functions such as endowing properties, acting as a carrier, enhancing the stability of drugs, solubilizing, cosolubilizing, sustained and controlled release, etc. when producing drugs or formulating prescriptions, such as humectants, solubilizers, lubricants, emulsifiers, antibacterial agents, solvents, antioxidants, etc. In some embodiments, the auxiliary ingredient is selected from one or more of humectants, lubricants, solubilizers, emulsifiers, humectants, antibacterial agents, solvents.

[0090] As used herein, the term "emulsifier" refers to a substance that can form a stable emulsion from a mixed liquid of two or more immiscible components (such as oil and water). In some embodiments, the emulsifier is cetyl alcohol.

[0091] As used herein, the term "humectant" refers to a substance that can attract or lock in moisture and help the skin retain moisture, which is crucial for maintaining the hydration state of the skin and preventing dryness. Dry skin is a common symptom often associated with psoriasis, and humectants are commonly used as a basic treatment regimen throughout the course of treatment. In some embodiments, the humectant is one or more of petrolatum, glycerol, and / or hexylene glycol; in some embodiments, three different humectants are used, wherein the first humectant is glycerol,

[0092] the second humectant is petrolatum, and the third humectant is hexylene glycol.

[0093] As used herein, the term "lubricant" refers to a substance that can reduce the friction of powders or particles during processing. In some embodiments, the lubricant is dimethicone.

[0094] As used herein, the term "solubilizer" refers to a surfactant that can increase the solubility of poorly soluble drugs in a solvent. By increasing the solubility of the drug, the content of the main drug in the preparation can be increased. In some embodiments, the solubilizer is PEG (polyethylene glycol).

[0095] As used herein, the term "antibacterial agent" refers to a substance that has inhibitory and killing effects on bacteria and other microorganisms. In some embodiments, the antibacterial agent is p - hydroxyacetophenone.

[0096] II. Examples

[0097] The present invention will be described in detail below through specific examples. It should be understood that the following examples are only for explanation and illustration, and do not limit the scope of the present invention in any form.

[0098] In the following embodiments, biochemical reagents not specifically described are conventional reagents in the art, which can be prepared according to conventional methods in the art or obtained commercially, and the specification is laboratory pure grade. Neo-A, CAS number: 630057-39-5, trade name: Neoprzewaquinone A, was purchased from Chengdu Push Bio-Technology Co., Ltd., China; 5% imiquimod was purchased from Sichuan Meixin Pharmaceutical Co., Ltd., China; 0.1% Protopic tacrolimus ointment was purchased from Astellas Pharma (China) Co., Ltd. Before the administration of Neo-A, it was prepared into a cream form for application.

[0099] Example 1: Preparation of Neo-A Cream

[0100] The preparation method of Neo-A cream is as follows:

[0101] (1) Mix water, PEG and glycerol and heat to 80 °C;

[0102] (2) Add petrolatum and cetyl alcohol in turn;

[0103] (3) Stir at 80 °C for 30 min;

[0104] (4) Stir and cool down to 50 °C and add p-hydroxyacetophenone;

[0105] (5) Stir dimethicone and Neo-A evenly at room temperature and add them when cooled down to 40 °C;

[0106] (6) Cool down to room temperature and add hexylene glycol.

[0107] The proportions of various auxiliary components in the cream are shown in Table 1 below:

[0108] Table 1. Preparation of Neo-A cream with different doses

[0109] Serial number Raw material name Percentage Purpose of use 1 Vaseline 13 Humectant 2 Dimethicone 4 Lubricant 3 PEG 2 Solubilizer 4 Cetyl alcohol 1.5 Emulsifier 5 Glycerol 3 Humectant 6 p-Hydroxyacetophenone 0.3 Antibacterial agent 7 Hexylene glycol 0.5 Humectant 8 Water Add up to 100 Solvent 9 Neo-A 0.54, 1.8 and 5.4 μM Therapeutic drug

[0110] Example 2: Experimental grouping and drug treatment

[0111] SPF-grade male BALB / c mice (7-9 weeks old, weighing 20-25 g) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. License number: SCXK (Jing) 2016-0011, Medical Laboratory Animal Qualification Certificate number: AEEI-2024-145. During the experiment, the mice were housed in a standard animal house at the Capital Medical University. The environmental settings in the animal house were a 12-hour day-night cycle, the room temperature was controlled at 24 ± 2 °C, and the relative humidity was maintained at 50 ± 10%. Sufficient water and food were provided. The animals were adaptively fed for 2-3 days before the experiment. All experiments were approved by the Animal Ethics Committee of the Capital Medical University.

[0112] BALB / C mice were randomly assigned to 8 groups to evaluate the efficacy of Neo-A alone and in combination with TACR in the prevention and treatment of psoriasis. The back skin of the mice was selected as the experimental observation area. One day before the experiment, the hair on the back of the mice was shaved off to expose a hairless area of 3 cm × 4 cm. 5% IMIQ at 75 mg / day was applied to the dorsal skin of the mice every day. The specific grouping and drug treatment are as follows (the specific implementation plan is shown in Table 2):

[0113] Control group - Only the blank cream (120 mg / day) was applied to the back skin of the mice every day to evaluate the effect of the blank cream on the back skin of the mice;

[0114] Neo-A group - Neo-A cream was applied to the back skin of the mice every day to evaluate the effect of Neo-A cream alone on the back skin of the mice;

[0115] Model group - Only 5% IMIQ at 75 mg / day was applied to the dorsal skin of the mice every day to evaluate the natural progression of the IMIQ-induced psoriasis model and to be used as a control for the drug treatment group;

[0116] The mice in the drug treatment group were given drug intervention 1 hour after IMIQ treatment every day, that is, the positive drug group (TACR) - TACR was applied to the external skin of the mice every day to explore the therapeutic effect of TACR treatment on improving psoriasis symptoms; the low, medium, and high dose groups of Neo-A (Neo-A-L, Neo-A-M, Neo-A-H) - Neo-A was applied to the external skin of the mice every day to explore the therapeutic effect of Neo-A treatment on improving psoriasis symptoms; the combined group (Neo-A+TCAR) - Neo-A+TCAR was applied to the external skin of the mice every day to explore the synergistic therapeutic effect when Neo-A and TCAR were used in combination.

[0117] The above were all administered continuously for 6 days. After applying the cream, the mice were placed in a cage without bedding for at least ten minutes to interrupt the licking behavior and let the cream dry. The experimental procedure is shown in Figure 1 and the experimental data obtained were all processed using Graphpad analysis software. Measurement data were expressed as mean ± SEM, and the non-parametric unpaired independent sample T-test method was used for data analysis, and the Mann Whitney test was used for comparison between groups. p<0.05 indicated significant differences.

[0118] Table 2. Experimental grouping and drug administration

[0119]

[0120] Example 3: Observation and evaluation results of psoriasis-like skin lesions in mice

[0121] During the process of modeling with IMIQ, the skin damage of mice was observed and recorded every day. According to the psoriasis severity index scoring standard, the erythema, scales and thickness of the mice's skin were scored at the experimental observation site. The scoring standard is shown in Table 3. The psoriasis severity scoring has a certain degree of subjectivity. To improve the scoring accuracy of this experiment, the skin lesions on the backs of the mice in each group were photographed and numbered every day. After removing the group information, random double-blind scoring was performed.

[0122] Table 3. Psoriasis Severity Index Scoring Standard

[0123]

[0124] The results of skin lesions showed that on the 2nd day of IMIQ modeling, erythema began to appear on the backs of the mice, initially light red. As the modeling time extended, the erythema, scales and skin thickening on the backs of the mice in the model group gradually worsened. By the 5th day of psoriasis modeling, large-scale desquamation and skin thickening began to appear on the backs of the mice in the model group, and the condition further deteriorated on the 7th day of modeling. Different drug treatment groups had varying degrees of alleviating effects ( Figure 2 A); Statistical analysis of the skin lesion severity index scores on the 7th day found that compared with the Control group, the skin lesion severity index scores of the mice in the model group increased significantly (p < 0.0001), while the groups given Neo-A (0.54, 1.8, 5.4 μM, 120 mg / day) and TACR (60 mg / day) all had varying degrees of improvement effects (p < 0.001, p < 0.01, p < 0.0001, p < 0.01); The combined drug treatment group of the high-dose TACR and Neo-A groups had significantly reduced skin lesion severity index scores compared with the model group (p < 0.0001), and the effect was significantly better than that of different doses of Neo-A (p < 0.001, p < 0.001, p < 0.001) and TACR (p < 0.0001) alone ( Figure 2 B and C), the erythema and scales disappeared, almost approaching normal skin; The above results indicate that these two treatments can effectively reduce the severity of psoriasis skin lesions, and the combined use of the high-dose TACR and Neo-A groups has an obvious synergistic effect.

[0125] Example 4: Detection Results of Histopathological Changes in Mouse Skin Lesions

[0126] On the 7th day of model establishment, the back skin tissues of the mice were taken for tissue fixation, paraffin embedding, sectioning, and HE staining. The tissues were observed under a microscope and scanned for photography for histopathological analysis. At the same time, the thickening of the spinous layer and the infiltration of inflammatory cells in the dermis were evaluated. The evaluation of spinous layer thickening was carried out through the following steps: measurement was performed under a 400-fold magnification field of view of the HE-stained microscope. The selected image was opened using Image J software. The pixel area of the epidermal region within this field of view was measured using the straight-line measurement tool. The length of the line was viewed in the software, which would represent the pixel length of the epidermal region, and the Baker score was performed.

[0127] The specific criteria for the Baker score are as follows:

[0128] The presence of Munro microabscesses in the epidermal layer is 2.0 points; hyperkeratosis is 0.5 points; parakeratosis is 1.0 points; thinning or disappearance of the granular layer is 1.0 points; thickening of the spinous layer is 1.0 points; elongation and undulation of the rete ridges are scored 0.5 points, 1.0 points, and 1.5 points according to mild, moderate, and severe degrees respectively. In the dermis, the infiltration of mononuclear or multinuclear cells is scored 0.5 points, 1.0 points, and 1.5 points according to mild, moderate, and severe degrees respectively; the upthrust of the papilla is 0.5 points; telangiectasia is 0.5 points.

[0129] The results of the Baker score showed that compared with the Control group, the mice in the model group showed pathological manifestations such as thickening of the epidermal spinous layer, hyperkeratosis or parakeratosis, upthrust of the papilla, and telangiectasia in the skin lesion area 6 days after model establishment (p<0.0001), while the groups treated with Neo-A (0.54, 1.8, 5.4 μM, 120 mg / day) and TACR (60 mg / day) all had varying degrees of improvement effects (p<0.05, p<0.01, p<0.001, p<0.0001); the combined treatment group of the high-dose TACR and Neo-A groups showed significantly reduced pathological phenotypes compared with the model group (p<0.001), and the effect was significantly better than that of different doses of Neo-A and TACR (p<0.001) alone ([ Figure 3 A and B); in addition, measurement of the back skin thickness found that compared with the Control group, the skin thickness of the mice in the model group increased significantly 7 days after model establishment (p<0.0001), while the groups treated with Neo-A (1.8, 5.4 μM, 120 mg / day) and TACR (60 mg / day) all had varying degrees of improvement effects (p<0.01, p<0.01, p<0.001); the combined treatment group of the high-dose TACR and Neo-A groups showed significantly reduced pathological phenotypes compared with the model group (p<0.001), and the effect was significantly better than that of different doses of Neo-A (p<0.01, p<0.01, p<0.01) and TACR (p<0.001) alone ([ Figure 3 C).

[0130] Example 5: Calculation of Mouse Spleen Index

[0131] We observed the body weight of each group of mice every day. After treatment with IMIQ (75 mg / day), the body weight of each group of mice showed a decrease compared with that of the Control and Neo-A (5.4 μM, 120 mg / day) alone. In addition to the effect of modeling on body weight, there were no significant changes in the body weight of the Neo-A (0.54, 1.8, 5.4 μM, 120 mg / day) and TACR (60 mg / day) groups ( Figure 2 A); On the 7th day, after sacrificing the mice, the intact spleens of the mice were carefully and quickly dissected. After drying the surface moisture or liquid, the spleen weight was weighed with a high-precision electronic balance, and the spleen index was calculated. Spleen index (%) = mouse spleen weight (mg) / mouse body weight (g).

[0132] As shown in the results, the spleen was significantly enlarged after IMIQ induction (p < 0.0001), while the combined treatment group of TACR and high-dose Neo-A was significantly reduced compared with it (p < 0.001, p < 0.01); after treatment with IMIQ, different doses of Neo-A were given, and there was no significant difference in the spleen index of mice; after treatment with high-dose Neo-A alone, there was no obvious effect on the spleen index of normal mice( Figure 2 B); The above results indicate that Neo-A has no toxic and immunosuppressive effects on mice.

[0133] The specific embodiments of the present invention have been described in detail above, but they are only examples, and the present invention is not limited to the specific embodiments described above. For those skilled in the art, any equivalent modifications and substitutions to the present invention are also within the scope of the present invention. Therefore, equivalent transformations and modifications made without departing from the spirit and scope of the present invention are all covered by the present invention.

Claims

1. Use of Neo-A in the preparation of a drug or a medicine kit for treating autoimmune related skin diseases.

2. The use according to claim 1, wherein the autoimmune-related skin disease comprises psoriasis, rheumatoid arthritis, systemic lupus erythematosus, atopic dermatitis, vitiligo, pemphigus or dermatomyositis; preferably psoriasis.

3. The use according to claim 1 or 2, wherein The medicament or kit further comprises tacrolimus (TACR); Preferably, the medicine or medicine kit comprises a pharmaceutical composition having Neo-A and / or TACR as an active ingredient.

4. The use according to claim 3, wherein The pharmaceutical composition further comprises pharmaceutically acceptable auxiliary ingredients; Preferably, the pharmaceutical composition is in the form of a topical application preparation, a solution preparation or more; More preferably, the pharmaceutical composition is in the form of a topical application preparation; More preferably, the pharmaceutical composition is in the form of a gel, cream or ointment.

5. The use according to claim 4, wherein The pharmaceutical composition is a topical smear preparation, and the auxiliary component is selected from one or more of a moisturizer, a lubricant, a solubilizer, an emulsifier, an antibacterial agent, and a solvent; Preferably, the moisturizer is petrolatum, glycerin and / or hexylene glycol; Preferably, the lubricant is dimethyl silicone oil; Preferably, the solubilizing agent is polyethylene glycol (PEG); Preferably, the emulsifier is cetyl alcohol; Preferably, the antibacterial agent is p-hydroxyacetophenone; Preferably, the solvent is water.

6. The use according to claim 5, wherein Calculated based on the total weight of the pharmaceutical composition, the weight percentage of the moisturizer is 10.0-20.0%, preferably 15.0-17.0%, more preferably 16.5%; Preferably, the weight percentage of the lubricant is 1.0-10.0%, preferably 2.0-6.0%, more preferably 4.0%, calculated based on the total weight of the pharmaceutical composition; Preferably, the weight percentage of the solubilizer is 1.0-5.0%, preferably 1.0-3.0%, more preferably 2.0%, calculated based on the total weight of the pharmaceutical composition; Preferably, the weight percentage of the emulsifier is 1.0-5.0%, preferably 1.0-3.0%, more preferably 1.5%, calculated based on the total weight of the pharmaceutical composition; Preferably, the weight percentage of the antibacterial agent is 0.05-1.0%, preferably 0.1-0.5%, more preferably 0.3%, calculated based on the total weight of the pharmaceutical composition; Preferably, the proportions of the components in the pharmaceutical composition, calculated based on the total weight of the composition, are: 16.5% moisturizer, 4% lubricant, 2% solubilizer, 1.5% emulsifier, and 0.3% antibacterial agent.

7. The use according to any one of claims 3 to 6, wherein The content of Neo-A in the pharmaceutical composition is 0.2-8.0 μM, preferably 0.54-5.4 μM; Preferably, the content of Neo-A is 0.54-1.0 μM, or 1.0-3.0 μM, or 4.0-5.4 μM; Preferably, the content of Neo-A is 0.54 μM; Preferably, the content of Neo-A is 1.8 μM; Preferably, the content of Neo-A is 5.4 μM.

8. The use according to any one of claims 3 to 7, wherein The dosage of the pharmaceutical composition product is 160-200 mg, preferably 180 mg.

9. The use according to any one of claims 3 to 8, wherein The pharmaceutical composition is prepared as follows: (1) mixing and heating a solvent, a humectant and a solubilizing agent; (2) adding moisturizer 2 and emulsifier in sequence; (3) Keep heating and stirring; (4) stirring, cooling, and adding an antibacterial agent; (5) Stir the lubricant and therapeutic drug evenly and add them after cooling; (6) Cooling and adding moisturizer 3; Preferably, the heating in step (1) and / or step (3) is heating to 75-85° C., preferably 80° C.; Preferably, the stirring in step (3) is for 20-40 min, preferably 30 min; Preferably, the cooling in step (4) is to 45-55°C, preferably 50°C; Preferably, the stirring in step (5) is stirring at room temperature; Preferably, the cooling in step (5) is to cool to 35-45°C, preferably 40°C; Preferably, the cooling in step (6) is cooling to room temperature; Preferably, the humectant is glycerin; Preferably, the second moisturizer is vaseline; Preferably, the moisturizing agent is hexylene glycol; Preferably, the lubricant is dimethyl silicone oil; Preferably, the solubilizing agent is polyethylene glycol (PEG); Preferably, the emulsifier is cetyl alcohol; Preferably, the antibacterial agent is p-hydroxyacetophenone; Preferably, the solvent is water; Preferably, the therapeutic drug is Neo-A; More preferably, the first moisturizer is glycerin, the second moisturizer is vaseline, the third moisturizer is hexylene glycol, the lubricant is dimethyl silicone oil, the solubilizer is PEG, the emulsifier is cetyl alcohol, the antibacterial agent is p-hydroxyacetophenone, and the solvent is water; More preferably, the therapeutic drug is Neo-A combined with TCAR.

10. A drug or kit for treating autoimmune-related skin diseases, comprising Neo-A and / or TACR; Preferably, the autoimmune-related skin disease comprises psoriasis, rheumatoid arthritis, systemic lupus erythematosus, atopic dermatitis, vitiligo, pemphigus or dermatomyositis; more preferably psoriasis.

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