Application of 6-amino-1H-pyrazolo [3, 4-d] pyrimidine JAK3 kinase inhibitor in treatment of atopic dermatitis

Abstract

The invention relates to the technical field of medicines, in particular to application of a JAK3 inhibitor N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazole-4-yl) amino)-1H-pyrazolo [3, 4-d] pyrimidine-1-yl) methyl) phenyl) acrylamide and pharmaceutically acceptable salts thereof in preparation of medicines, and a medicine for treating atopic dermatitis. According to the embodiment of the invention, an atopic dermatitis mouse model is established, a JAK3 inhibitor is orally administered to the mouse, the skin lesion of the mouse is reduced, and further research finds that the pathological injury of the mouse is reduced, the spleen index of the mouse is reduced, the IgE concentration in the blood of the mouse and the secretion of inflammatory factors such as IL-4 in skin lesion tissues are reduced, which indicates that the JAK3 inhibitor inhibits the occurrence of allergic reaction, and the skin lesion of the mouse is reduced. The injury of immune response to the skin is relieved, so that the skin state is improved, and a safe and effective treatment effect on the skin lesion caused by atopic dermatitis is achieved.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and is characterized by use of N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)acrylamide and a pharmaceutically acceptable salt thereof as an active ingredient in the preparation of a drug for treating atopic dermatitis, and also relates to a drug for treating atopic dermatitis. Background Art

[0002] Atopic dermatitis (AD) is a chronic, recurrent, inflammatory skin disease. Most AD patients develop the disease in infancy, and severe cases may even persist into adulthood. The recurrence of AD seriously affects the quality of life of patients. The onset of AD is closely related to genetic and environmental factors. It is generally believed that on the basis of genetic factors, patients are exposed to allergens, which leads to abnormal skin immune reactions and inflammation, causing rashes and itching. There are multiple abnormal immune response links in the occurrence and development of AD, such as: presentation of allergens by Langerhans cells and skin dendritic cells, abnormal immune response dominated by Th2, dysfunction of regulatory T cells, excessive production of IgE and increased eosinophils.

[0003] At present, the symptoms of AD are mainly relieved by topical drugs such as glucocorticoids (TCS), calcineurin inhibitors (TCI), PDE4 inhibitors, antihistamines, and immunosuppressants. AD chooses different treatment options according to the severity of the disease. Mild patients can be treated with topical calcineurin inhibitors, glucocorticoids, etc.; moderate patients are recommended to continue to use TCS / TCI for active maintenance treatment after using topical drugs to control skin lesions, and oral antihistamines and anti-infective drugs or combined with phototherapy when necessary; severe patients should consider using systemic immunosuppressants (for example, cyclosporine, methotrexate), biological agents, and phototherapy. In the treatment methods for moderate to severe AD, both topical and systemic treatments have certain limitations and adverse reactions. For example, long-term and large-area use of TCS may cause skin and systemic adverse reactions; TCI may cause local burning and irritation; systemic immunosuppressants may cause leukopenia and abnormal liver and kidney function; systemic glucocorticoids may cause electrolyte disorders, infection and other adverse reactions, and the long-term efficacy is limited, and recurrence or rebound may occur after discontinuation of medication. Therefore, there is still a need for a drug or treatment method that does not cause side effects to patients, can also be used to combat the onset of allergies and the development of skin inflammation, and can be applied to various categories of patients.

[0004] Janus kinase-signal transducer and activator of transcription (JAK-STAT) pathway signals are closely related to the onset of AD. JAK kinases are involved in regulating multiple immune inflammatory pathways in the course of AD, including Th2, Th22, Th1, and Th17 responses involved in the onset of AD. Importantly, cytokines IL-4, IL-13, IL-31, and IL-22 require JAK-STAT downstream signals to exert their biological functions. JAK inhibitors inhibit AD inflammation and itch-related molecular signaling pathways through multiple targets, which can quickly relieve itching and improve skin lesions. They can be used as early rapid induction of remission treatment during the onset of the disease or as a rescue treatment during the exacerbation period. Highly selective small molecule inhibitors of JAK3 have obvious advantages in reducing adverse reactions compared to non-selective JAK inhibitors and other immunosuppressants. Another major advantage of JAK inhibitor drug therapy is that it can be applied topically.

[0005] N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)acrylamide is a highly selective small molecule inhibitor of JAK3, which inhibits JAK3 phosphorylation with IC 50 The selectivity of JAK3 to other isoforms is more than 4500 times, showing a very high selectivity. Studies on signaling pathways at the cellular level have shown that N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)acrylamide only has a clear inhibitory effect on STAT phosphorylation induced by six γc cytokines regulated by the JAK3 subtype, but has no inhibitory effect on STAT phosphorylation induced by other subtype-related cytokines, further confirming the highly selective inhibition of JAK3 by N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)acrylamide (Chen CJ, et al. Science advances, 2022, 8(33):eabo4363).

[0006] Currently, there is no report on the JAK3 inhibitor N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)acrylamide in the treatment of atopic dermatitis. Summary of the invention

[0007] The present invention aims to provide a new medical use of N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)acrylamide represented by general formula I, specifically, to provide N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]

[0008] The invention discloses use of (1-pyrimidin-1-yl)methyl)phenyl)acrylamide and pharmaceutically acceptable salts thereof in preparing a drug for treating atopic dermatitis, and provides a drug for treating atopic dermatitis.

[0009]

[0010] In a first aspect, the present invention provides an application of a pharmaceutical composition in the preparation of a drug for preventing and / or treating atopic dermatitis, wherein the pharmaceutical composition is the application of N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)acrylamide and a pharmaceutically acceptable salt thereof as shown in Formula I in the preparation of a drug for treating atopic dermatitis.

[0011] Furthermore, in the above-mentioned use, the above-mentioned medicine is a medicine for alleviating skin lesions caused by atopic dermatitis.

[0012] Furthermore, in the above-mentioned use, the above-mentioned medicine is a medicine for enhancing the skin barrier function.

[0013] Furthermore, in the use, the drug is a drug for inhibiting the secretion of IgE and cytokines, wherein the cytokines include one or more of IL-4 and IL-13.

[0014] In a second aspect, the present invention provides a drug for treating atopic dermatitis, comprising N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)acrylamide and a pharmaceutically acceptable salt thereof.

[0015] Furthermore, the drug is a clinically acceptable preparation made from N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)acrylamide and a pharmaceutically acceptable salt thereof.

[0016] Furthermore, the oral preparations include solid powders, tablets, dispersible granules, capsules, and pills; preferably, the powders and tablets contain about 0.1% to about 99.9% of active ingredients; solid excipients are selected from magnesium carbonate, magnesium stearate, talc, sugar or lactose; liquid preparations include solutions, suspensions and emulsions, preferably, the liquid is an aqueous solution or a water-propylene glycol solution for parenteral injection, or an oral solution with added sweeteners and contrast agents.

[0017] Furthermore, the preparation is an external preparation, including an external ointment, a liniment, a film or an external spray preparation.

[0018] In summary, compared with the prior art, the present invention achieves the following technical effects:

[0019] The present invention adopts a classic atopic dermatitis mouse modeling method to replicate an oxazolone-induced atopic dermatitis model mouse, and administers N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)acrylamide to the model mice for intervention treatment. The results show that N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)acrylamide has a significant anti-atopic dermatitis effect and can be used to prepare a drug for treating atopic dermatitis. The present invention expands the medical use of the JAK3 inhibitor N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)acrylamide, and provides a new direction for the treatment of atopic dermatitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0021] Figure 1 The weight of each group of mice in the back stimulation mouse model of AD model in Experimental Example 1.

[0022] Figure 2 These are representative photos of the skin lesions on AD model mice stimulated on the back in Experimental Example 1.

[0023] Figure 3 These are the results of pathological examination of the skin lesions of AD mice in Experimental Example 1.

[0024] Figure 4 The spleen index results of AD model mice stimulated on the back in Experimental Example 1. ## =(P<0.01) compared with the blank control group mice, ***(P<0.001) compared with the model control group mice.

[0025] Figure 5 This is a photo of the lymph nodes of mice stimulated on the back of the AD model in Experimental Example 1.

[0026] Figure 6 The results of IgE, IL-4, and IL-13 detection in the blood and skin lesions of mice in Experimental Example 1: (a) IgE and IL-4 levels in mouse serum; (b) IgE and IL-4, IL-13 expression levels in the back skin of mice. #### Compared with the blank control group mice, P<0.0001. ### =(P<0.001) compared with the blank control group mice, **** represents P<0.0001 compared with the model control group mice, *** represents P<0.001 compared with the model control group mice, and ** represents P<0.01 compared with the model control group mice).

[0027] Figure 7 This is the body weight of AD mice stimulated by ear in Experimental Example 2.

[0028] Figure 8 Representative photos of the ears of the mouse AD model subjected to ear stimulation in Experimental Example 2.

[0029] Fig. 9 is the ear thickness of the mouse AD model in Experimental Example 2.

[0030] Fig.10 These are the results of pathological examination of mouse ear pieces in Experimental Example 2.

[0031] Fig.11 This is a photo of the cervical draining lymph nodes of AD mice after ear stimulation in Experimental Example 2.

[0032] Fig.12 This is the result of IgE detection in the blood of AD mice stimulated by ear in Experimental Example 2. #### =(P<0.0001) compared with the blank control group mice, **** represents P<0.0001 compared with the model control group mice, *** represents P<0.001 compared with the model control group mice, ** represents P<0.01 compared with the model control group mice, and * represents P<0.05 compared with the model control group mice). DETAILED DESCRIPTION

[0033] The following experimental examples are provided for a better understanding of the present invention, but are not limited to the best implementation mode and do not limit the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior arts shall fall within the protection scope of the present invention.

[0034] The medicine provided herein can be formulated into an oral dosage form suitable for producing a systemic effect, and the oral dosage provided herein can be provided in a solid, semisolid or liquid dosage form for oral administration. Oral administration also includes buccal administration, tongue or sublingual administration. Suitable oral dosage forms include but are not limited to tablets, instant tablets, chewable tablets, capsules, pills, strips, lozenges, lozenges, pastilles, cachets, granules, medical chewing gums, bulk powders, effervescent or non-effervescent powders or granules, oral aerosols, emulsions, suspensions, wafers, powders, elixirs and syrups. The medicine may contain one or more pharmaceutically acceptable excipients, including but not limited to adhesives, fillers, diluents, disintegrants, wetting agents, lubricants, glidants, colorants, dye migration inhibitors, sweeteners, flavoring agents, emulsifiers, suspensions and dispersants, preservatives, solvents, non-aqueous liquids, organic acids and carbon dioxide sources.

[0035] The agents provided herein can be formulated into any dosage form suitable for topical administration to produce a local effect, including emulsions, creams, gels, hydrogels, ointments, powders, lotions, suspensions, tinctures, pastes, foams, films, aerosols, rinses, sprays, suppositories, bandages and skin patches.

[0036] Example 1. Pharmacodynamic study of JAK3 selective inhibitor on back stimulation AD model

[0037] 1. Experimental Materials

[0038] 1.1 Experimental animals

[0039] 6-8 weeks old SPF female BALB / c mice, 18-20 g, were purchased from Beijing Huafukang Biotechnology Co., Ltd., certificate number: No.110322221103000916.

[0040] 1.2 Reagents and consumables

[0041] Test substance: N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)acrylamide; Modeling agent: oxazolone.

[0042] IL-4, IL-13, IgE ELISA kits, mouse IL-4 immunoassay kit (ProQuantum) were purchased from Thermo Fisher; Minute skin total protein extraction kit.

[0043] If no specific experimental steps or conditions are specified in the experimental examples, the conventional experimental steps or conditions described in the literature in the field can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be obtained commercially.

[0044] The N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)acrylamide of the present invention has a structure as shown in Formula I:

[0045]

[0046] 2. Experimental Methods

[0047] 2.1 Experimental Grouping

[0048] BALB / c mice were housed in the Experimental Animal Center of the Institute of Materia Medica, Chinese Academy of Medical Sciences, with free access to food and water, room temperature of 22-25°C, relative humidity of 55-65%, and 12h light cycle. All animal experiments were strictly carried out in accordance with the guidelines of the Experimental Animal Ethics Committee of the Chinese Academy of Medical Sciences and Peking Union Medical College and approved by the Ethics Committee. Thirty experimental mice were randomly divided into 5 groups, 6 mice in each group, namely blank control, model control group, high, medium and low dose groups of compound (I).

[0049] 2.2 Establishment of AD mouse model

[0050] On day 0 of the experiment, the hair on the back of the mice was shaved, with an area of ​​2.5 cm × 2.5 cm. On days 1 and 2 of the experiment, the skin of the mice in the model group was sensitized and stimulated, and the depilated area on the back of the mice was smeared once with 50 μL of a mixed solution of 5% oxazolone in acetone / soybean oil (4:1). Starting from day 7, 50 μL of 0.5% oxazolone was applied to the back of the model group and the drug-treated group to induce AD-like lesions once every two days for three weeks (10 times). AD manifestations such as itching, erythema, papules, and scabs appeared on the skin in the depilated area of ​​the mice, and the model was successfully established.

[0051] 2.3 Administration

[0052] The dosage settings of the administration groups are as follows: high dose of compound (I) (30 mg / kg), medium dose of compound (I) (10 mg / kg), and low dose of compound (I) (3 mg / kg).

[0053] Drug preparation: Prepare suspension of corresponding concentration with 0.5% CMC-Na.

[0054] From the first day of the experiment, each group of model mice was given oral administration, and the normal control group and model control group were given an equal volume of solvent once a day. The oral administration volume of mice was 10 μL / g.

[0055] 3. Evaluation indicators

[0056] 3.1 Weight

[0057] From day 0 to the end of the experiment, the body weight of mice in each group was measured every 2 days.

[0058] 3.2 Skin lesions

[0059] The back skin lesions of mice in each group were observed and recorded, and the lesions were photographed every 2 days. The back skin lesions were scored according to the severity of erythema / hemorrhage, edema, scaling and lichenification, with each sign scored as 0 (none), 1 (mild), 2 (moderate) and 3 (severe).

[0060] 3.3 Spleen index and lymph node size

[0061] After killing, the spleen of the mouse was separated, the surface moisture was dried, and the weight was recorded according to the formula: spleen index =

[0062] The spleen weight (mg) / mouse body weight (g) was calculated. The inguinal lymph nodes of mice were separated and photographed to observe the size of lymph nodes in each group.

[0063] 3.4 Determination of cytokine levels in serum

[0064] 24 hours after the last sensitization, the mice were anesthetized, the right eyeball was removed and blood was collected, and serum was applied

[0065] Elisa kit was used to detect the cytokine content in serum.

[0066] 3.5 Pathological examination of skin lesions

[0067] At the end of the experiment, the corresponding skin lesion tissues were cut from each group, fixed in 10% formaldehyde solution, embedded in paraffin, and stained with HE.

[0068] 4. Statistical Analysis

[0069] The experimental data are expressed as Mean ± SEM. Continuous data were compared using analysis of variance (ANOVA) followed by Duncan's test; discrete data were compared using nonparametric tests such as Kruscal-Wallis ANOVA and

[0070] Median Test was used to analyze the experimental results and present them in graphs using GraphPad Prism version 8.0 software.

[0071] 5. Experimental Results

[0072] 5.1 Animal weight

[0073] Figure 1 The results showed that the compound (I) groups had no significant effect on the body weight of mice compared with the model group during the administration period, indicating that the mice had good tolerance to oral administration of compound (I) and no significant changes in body weight were observed due to administration. That is, oral administration of 30 mg / kg of compound (I) had no toxic side effects on mice.

[0074] 5.2 Skin lesion symptoms

[0075] like Figure 2 As shown, oral administration of JAK3 selective inhibitor compound (I) at the end of the test (after ten stimulations) can effectively alleviate the atopic dermatitis-like symptoms of mice induced by oxazolone. The backs of the model group and each drug-treated group showed different degrees of inflammatory reactions, while the normal control group did not show inflammatory reactions. Under a magnifying glass, the skin of the normal control group was ruddy, without erythema, papules and other inflammatory reactions; the skin of the model group was obviously swollen and raised, with more papules on the erythema, erosion surface, and yellow scabs; the skin of the compound (I) group was mildly flushed, with a small amount of light red papules, and no obvious erosion or scabs.

[0076] Compound (I) alleviated the dermatitis symptoms of erythema, edema, scaling and lichenification on the back of mice in a dose-dependent manner.

[0077] Table 1 Scoring of clinical features of atopic dermatitis: scaling, lichenification, edema, and erythema.

[0078]

[0079]

[0080]

[0081]

[0082]

[0083] #### represents P<0.0001 compared with the blank control group mice), **** represents P<0.0001 compared with the model control group mice

[0084] 5.3 Pathological examination of skin lesions

[0085] Histopathological results showed that the model group mice had epidermal hyperplasia, thickening of the spinous cell layer, tissue edema and a large number of inflammatory cells infiltrating the dermis. Compound (I) reduced the histopathological changes in a dose-dependent manner. Figure 3 It can be seen that the inflammatory cells in the high-dose and medium-dose groups of compound (I) were reduced, the structural integrity of cells in each layer of the dermis was improved, and the pathological damage was significantly alleviated.

[0086] 5.4 Immunosuppressive activity detection

[0087] The spleen index results are shown in Figure 4. Compared with the normal control group, the spleen index of the atopic dermatitis model mice was significantly higher than that of the normal control group mice ( ## P<0.01), indicating that the atopic dermatitis model mice were successfully established; compared with the model group, the spleen index of the mice in the high-dose group of compound (I) was significantly lower than that in the model group (***P<0.001), indicating that compound (I) can significantly reduce the spleen index of atopic dermatitis model mice, thereby indicating that compound (I) has a significant immunosuppressive effect. Figure 5 The compound (I) also has a certain inhibitory effect on lymph node enlargement in dermatitis mice.

[0088] 5.5 IgE and cytokine levels in serum and skin lesions

[0089] The concentrations of IL-4 and Ig E in serum were determined by enzyme-linked immunosorbent assay. Figure 6 The JAK3 selective inhibitor compound (I) can inhibit the level of Th2-related cytokines and IgE secretion. Compared with the normal control group, the concentrations of IL-4 and IgE in the serum of atopic dermatitis model mice were significantly higher than those in the normal control group mice ( #### P<0.0001), indicating that the atopic dermatitis model mice were successfully established; compared with the model group, the concentrations of IL-4 and Ig E in the serum of mice in each compound (I) administration group were significantly lower than those in the model group (***P

[0090] <0.001), which shows that compound (I) can significantly reduce the concentration of Ig E in the serum of atopic dermatitis model mice, that is, compound (I) has a significant anti-atopic dermatitis effect. At the same time, the IgE level in the skin lesions of mice treated with compound (I) also decreased significantly; in addition, compound (I) also effectively inhibited the expression of Th2 type factors IL-4 and IL-13 in the skin lesions.

[0091] Example 2: Pharmacodynamic study of JAK3 selective inhibitors on ear stimulation AD model

[0092] 1. Experimental Materials

[0093] Same as Example 1

[0094] 2. Experimental Methods

[0095] 2.1 Experimental Grouping

[0096] Same as Example 1

[0097] 2.2 Model establishment

[0098] On the first and second days of the experiment, the abdomen and right ears of the model group mice were sensitized and stimulated, and a mixed solution of 5% oxazolone in acetone / soybean oil (4:1) was applied to the abdomen and the inside and outside of the right ear, with a dosage of 50 μL and 20 μL, respectively. Starting from the seventh day, 20 μL of 0.2% oxazolone was applied to the inside and outside of the right ear in the model group and the test drug group to induce AD-like lesions, once every two days for three weeks (10 times).

[0099] 2.3 Administration

[0100] Same as Example 1

[0101] 3. Evaluation indicators

[0102] 3.1 Weight

[0103] From day 0 to the end of the experiment, the body weight of mice in each group was measured every 2 days.

[0104] 3.2AD Performance

[0105] Ear swelling: The thickness of the ear was measured with a thickness gauge 24 hours after each application of oxazolidinone to the right ear.

[0106] 3.3 Lymph node size

[0107] The draining lymph nodes on the right ear of the mice were separated, and photos were taken to observe the size of the lymph nodes in each group.

[0108] 3.4 Determination of cytokine levels in serum

[0109] Same as Example 1

[0110] 3.5 Pathological examination of skin lesions

[0111] At the end of the experiment, the right auricle of each group was cut, fixed in 10% formaldehyde solution, embedded in paraffin, and stained with HE.

[0112] 4. Statistical Analysis

[0113] Same as Example 1

[0114] 5. Experimental Results

[0115] 5.1 Animal weight

[0116] Figure 7The results showed that the compound (I) groups had no significant effect on the body weight of mice compared with the model group during the administration period, indicating that the mice had good tolerance to oral administration of compound (I) and no significant changes in body weight were observed due to administration. That is, oral administration of 30 mg / kg of compound (I) had no toxic side effects on mice.

[0117] 5.2 Symptoms of ear swelling

[0118] like Figure 8 The compound (I) improved the wrinkling or desquamation of the ear skin of mice in a dose-dependent manner, and the redness and swelling of the ear skin were significantly improved compared with the model group. Fig. 9 As shown. Compared with the normal control group, the ear swelling of the atopic dermatitis model mice was obvious, indicating that the atopic dermatitis model mice were successfully modeled; compared with the model group, the ear swelling of the mice in each group of compound (I) was significantly lower than that in the model group. It shows that compound (I) can significantly reduce the ear swelling of the atopic dermatitis model mice, thereby indicating that compound (I) has a significant anti-atopic dermatitis effect.

[0119] 5.3 Pathological examination of skin lesions

[0120] like Fig.10 The histopathological results of the ear stimulation groups all showed that the model group mice had epidermal hyperplasia, thickening of the spinous cell layer, tissue edema and a large number of inflammatory cells infiltrating the dermis. Compound (I) reduced the severity of histopathological changes in a dose-dependent manner.

[0121] 5.4 Lymph node size

[0122] like Fig.11 The compound (I) also has a certain inhibitory effect on lymph node enlargement in dermatitis mice, which indicates that the compound (I) has a significant immunosuppressive effect.

[0123] 5.5 IgE and cytokine levels in serum and skin lesions

[0124] like Fig.12 The JAK3 selective inhibitor compound (I) can inhibit IgE secretion. Compared with the model group, the concentration of Ig E in the serum of mice in each compound (I) administration group was significantly lower than that in the model group, which shows that compound (I) can significantly reduce the concentration of IgE in the serum of atopic dermatitis model mice, that is, compound (I) has a significant anti-atopic dermatitis effect.

[0125] The experimental example of the present invention established an atopic dermatitis mouse model, and found that the skin lesion area of ​​the model mice administered with compound (I) was significantly reduced. Further studies found that compound (I) can significantly reduce the pathological damage of mice, inhibit the secretion of cytokines such as IgE in the blood of mice and IL-4 in the skin lesion tissue, thereby improving the skin condition, enhancing the skin barrier function, inhibiting the occurrence of allergic reactions, alleviating skin damage caused by immune response, and playing a safe and effective therapeutic role on the skin lesions caused by atopic dermatitis.

[0126] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. Use of N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)acrylamide as shown in formula I or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating atopic dermatitis, 2. Use of a pharmaceutical composition in the preparation of a drug for preventing and / or treating atopic dermatitis, characterized in that: The pharmaceutical composition is made of N-(3-((6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)acrylamide shown in formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

3. The use according to claim 2, characterized in that The preparation of the pharmaceutical composition is an oral preparation.

4. The use according to claim 3, characterized in that The oral preparations include solid powders, tablets, dispersible granules, capsules, and pills; preferably, the powders and tablets contain about 0.1% to about 99.9% of active ingredients; solid excipients are selected from magnesium carbonate, magnesium stearate, talcum powder, sugar or lactose; liquid preparations include solutions, suspensions and emulsions, preferably, the liquid is an aqueous solution or a water-propylene glycol solution for parenteral injection, or an oral solution with added sweeteners and contrast agents.

5. The use according to claim 2, characterized in that The pharmaceutical composition is in the form of a topical preparation.

6. Use according to claim 5, characterized in that The topical preparation is a spray preparation of a cream, an ointment, a gel, an emulsion, a film, or an external solution or suspension.

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