New use of FN-1501 in the preparation of drugs for treating and preventing autoimmune diseases

Abstract

The present invention discloses a new use of FN-1501 in the preparation of drugs for treating and preventing autoimmune diseases. Experiments show that the compound FN-1501 can significantly inhibit Th17 cell differentiation in vitro and alleviate many symptoms of psoriasis and rheumatoid arthritis mouse models. FN-1501 has good application prospects in the preparation of drugs for preventing and treating autoimmune diseases such as psoriasis and rheumatoid arthritis.

Description

Technical Field

[0001] The present invention relates to a new use of a compound, and in particular to a new use of FN-1501 in preparing a drug for treating and preventing autoimmune diseases. Background Art

[0002] Autoimmune diseases refer to diseases caused by abnormal immune responses to self-antigens. The pathogenesis of autoimmune diseases mainly involves the effects of cytokines, chemokines, adhesion molecules and immune cells. Helper T cells 17 (Th17) are associated with the pathogenesis of most autoimmune diseases, including psoriasis, rheumatoid arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, etc. Th17 cells are the third type of helper T cells in the Th family. They induce inflammation and damage by producing signature cytokines such as IL-17A, IL-17F and IL-22, and thus participate in the pathogenesis of autoimmune diseases. Some biologics targeting Th17 cell effector cytokines have been approved for the treatment of autoimmune diseases.

[0003] Th17 cells and their mediated signaling pathways play a key role in autoimmune diseases such as psoriasis, rheumatoid arthritis, and multiple sclerosis. The cytokines secreted by Thv7 cells are the main pathogenic factors that promote abnormal differentiation and proliferation of keratinocytes, and can participate in the induction of the formation of skin pathological plaques and damage to the skin barrier. At the same time, clinical data show that the number of pathogenic Th17 cells in peripheral blood mononuclear cells of patients with rheumatoid arthritis is significantly increased. IL-17 secreted by Th17 cells induces osteoclast growth, angiogenesis, and recruitment of innate immune cells to diseased joints, further aggravating the inflammatory response. Similarly, Th17 cells have also been identified as a key pathogenic cell population in multiple sclerosis. Anti-IL-17 monoclonal antibodies can effectively inhibit the pathogenesis of multiple sclerosis. Therefore, targeting Th17 cells and their mediated signaling pathways may be an effective strategy for the treatment of various autoimmune diseases such as psoriasis, rheumatoid arthritis, and multiple sclerosis.

[0004] FN-1501(C 22 H 25 N 9 FN-1501 (CAS No.: 1429515-59-2) is a small molecule compound jointly developed by China Pharmaceutical University and Shanghai Fosun Pharmaceutical Industry Development Co., Ltd., which has anti-tumor activity. Currently, there is no functional research on FN-1501 in the treatment and prevention of Th17 cells involved in autoimmune diseases. Summary of the invention

[0005] Purpose of the invention: The present invention provides a new use of FN-1501 in the preparation of drugs for treating and preventing autoimmune diseases such as psoriasis, rheumatoid arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, etc. Specifically, the drug of the present invention is made of FN-1501 as a single ingredient; or FN-1501 as an active ingredient, plus pharmaceutically acceptable excipients and salt forms or crystal forms; the pharmaceutically acceptable salt form is one or more of chloride, phosphate, sulfate, nitrate, nitrite, phosphate, citrate, tartrate, fumarate, mesylate, and maleate. The pharmaceutically acceptable crystal form is the crystal form corresponding to the free base. The drug dosage form is tablets, capsules, granules, powders, powder injections, pills, emulsions, suspensions, solutions, sprays, patches or sublingual tablets, etc.; the FN-1501 has the effect of inhibiting Th17 cell differentiation; the autoimmune disease is characterized by pathogenic cell subpopulations containing Th17 cells; the autoimmune disease is psoriasis, rheumatoid arthritis, multiple sclerosis, inflammatory bowel disease, ankylosing spondylitis, etc.; the use of the above-mentioned pharmaceutical composition in the preparation of drugs for treating and preventing autoimmune diseases is also within the protection scope of the present invention.

[0006] Technical solution: Use of FN-1501 in the preparation of drugs for treating and preventing autoimmune diseases.

[0007] The use described herein, the autoimmune diseases include psoriasis, rheumatoid arthritis, multiple sclerosis, inflammatory bowel disease, and ankylosing spondylitis.

[0008] The CAS number of the FN-1501 is: 1429515-59-2.

[0009] The invention relates to the use of a pharmaceutical composition in preparing a drug for treating and preventing autoimmune diseases; the pharmaceutical composition is prepared by using FN-1501 as an active ingredient and adding pharmaceutically acceptable excipients and salt forms, crystal forms or pharmaceutically acceptable carriers.

[0010] The pharmaceutically acceptable salt type is one or more of chloride, phosphate, sulfate, nitrate, nitrite, phosphate, citrate, tartrate, fumarate, methanesulfonate, and maleate.

[0011] The use described, the pharmaceutically acceptable crystal form is the crystal form corresponding to the free base.

[0012] The use described herein, the autoimmune diseases include psoriasis, rheumatoid arthritis, multiple sclerosis, inflammatory bowel disease, and ankylosing spondylitis.

[0013] The CAS number of the FN-1501 is: 1429515-59-2.

[0014] The use described, the dosage form of the pharmaceutical composition includes tablets, capsules, granules, powders, powder injections, pills, emulsions, suspensions, solutions, sprays, patches or sublingual tablets.

[0015] The pharmaceutically acceptable excipients include diluents, adhesives, disintegrants, glidants, lubricants, flavoring agents, inclusion materials, and adsorption materials.

[0016] In the described use, FN-1501 targets helper T cells 17 and the signaling pathways mediated by them.

[0017] In the above-mentioned use, FN-1501 has the effect of inhibiting Th17 cell differentiation.

[0018] The experiment of the present invention shows that FN-1501 has a significant inhibitory effect on Th17 cell differentiation in vitro, and reduces the mRNA expression levels of transcription factors and inflammatory factors such as IL-17A, IL-17F, IL-22, RORγt, IL-6, and TNFα. In the imiquimod-induced psoriasis mouse model, FN-1501 can effectively alleviate obvious psoriasis-like lesions such as erythema, skin thickening, and desquamation on the back of the mouse skin, and significantly improve epidermal hyperplasia and inflammatory cell infiltration. In the collagen-induced mouse arthritis model, FN-1501 can significantly reduce the ankle clinical score index of mice, inhibit synovial inflammation and fibrous tissue hyperplasia, cartilage and bone damage in mice. In summary, FN-1501 can be used to treat autoimmune diseases such as psoriasis and rheumatoid arthritis by inhibiting Th17 cell differentiation. FN-1501 has application value in the clinical treatment and prevention of autoimmune diseases related to Th17 cell differentiation.

[0019] The key technical points of the present invention are: 1. FN-1501 has a significant inhibitory effect on Th17 cell differentiation; 2. FN-1501 can be used to prepare drugs for treating and preventing autoimmune diseases such as psoriasis and rheumatoid arthritis.

[0020] Beneficial effects: Compared with the prior art, the compound of the present invention, FN-1501, can be used as an active ingredient in drugs for treating and preventing autoimmune diseases, providing a new solution for the treatment of autoimmune diseases. Experiments have shown that the compound FN-1501 can significantly inhibit Th17 cell differentiation in vitro and alleviate many symptoms of psoriasis and rheumatoid arthritis mouse models. FN-1501 has good application prospects in the preparation of drugs for preventing and treating autoimmune diseases such as psoriasis and rheumatoid arthritis. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 FN-1501 inhibits the differentiation of Th17 cells in vitro;

[0022] Figure 2 FN-1501 inhibits the mRNA expression of Th17 cell-related transcription factors and inflammatory factors;

[0023] Figure 3 FN-1501 improves imiquimod-induced psoriasis-like lesions in mouse skin;

[0024] Figure 4 FN-1501 reduces the PASI score in the imiquimod-induced psoriasis mouse model;

[0025] Figure 5 This is the result of hematoxylin-eosin staining (HE staining) of mouse skin tissue sections;

[0026] Figure 6 FN-1501 improves joint swelling in mice with collagen-induced arthritis;

[0027] Figure 7 This is the HE staining result of mouse ankle joint tissue. DETAILED DESCRIPTION

[0028] The structural formula of FN-1501 is:

[0029]

[0030] Example 1: FN-1501 inhibits Th17 cell differentiation in vitro

[0031] Female C57BL / 6J mice, 6-8 weeks old, weighing 18-22g, were given free access to water and food, with a 12h light-dark cycle per day and an ambient temperature of 25±2℃. The mice were euthanized and the spleens were removed. The spleen tissue was washed three times with PBS solution containing 10% double antibody, then transferred to a 70μm cell sieve and ground into a single cell suspension. Centrifuged at 1500rpm for 5min, and the supernatant was discarded. CD4 was enriched by immunomagnetic beads + T cells. The cells in the Th0 group were resuspended in IMDM complete medium, and the cells in the Th17 group and FN-1501 group were resuspended in Th17 differentiation medium (IMDM complete medium, containing IL-6, IL-23, TGF-B, anti-IFN-γ, anti-IL-4, anti-CD28). At the same time, 0.1 μM FN-1501 was added to the cells in the FN-1501 group, and no intervention was performed in the Th17 group. Finally, the cells were inoculated in a cell culture plate pre-coated with anti-CD3 and incubated at 37°C and 5% CO 2Cultured in an incubator for 72 hours. The cells were stimulated with IMDM complete medium containing intracellular stimulation reagents, then stained with membrane marker antibody CD4-FITC, fixed and permeated with IL-17A-APC for intracellular staining, and flow cytometry was performed. Alternatively, cells were directly collected without staining and the mRNA expression levels of Th17 cell-related transcription factors and inflammatory factors were detected using real-time fluorescence quantitative PCR (RT-qPCR) technology.

[0032] The results are as follows Figure 1 As shown in the flow cytometry results, FN-1501 significantly reduced CD4 + IL-17A + cell ratio, inhibiting the differentiation of Th17 cells in vitro.

[0033] like Figure 2 As shown, FN-1501 can inhibit Th17 cell differentiation in vitro and inhibit the mRNA expression of Th17 cell-related transcription factors and inflammatory factors. The results are expressed as Mean ± SEM, and one-way ANOVA was used for variance analysis. Compared with the Th0 group, ## P<0.01, ### P<0.001; compared with Th17 group, *** P<0.001.

[0034] Example 2: Pharmacological study of FN-1501 in a mouse psoriasis model

[0035] Female C57BL / 6J mice, 6-8 weeks old, weighing 18-22g, were given free water and food, with a 12h light-dark cycle per day and an ambient temperature of 25±2℃. Adaptive feeding was performed for 2 weeks before the start of the study. The mice were randomly divided into 3 groups: normal control group (Control), model control group (Model), and FN-1501 group (FN-1501). The day before modeling, the back of the mice was shaved to create a bare area of ​​about 2cm*3cm on the skin. In the Model and FN-1501 groups, a mouse psoriasis model was established by applying 5% imiquimod cream 62.5mg / mouse / day on the back of the mice for 7 days; the Control group was applied with an equal amount of vaseline. The FN-1501 group was given FN-1501 (2.4mg / kg, iv) on days 1, 3, and 5. The erythema, scaling, and skin thickness of the back skin of the mice were observed every day, and the PASI score was calculated. The specific scoring criteria are shown in Table 1. After the modeling and drug administration, photos of the back skin of the mice were taken, and then the mice were euthanized and the back skin tissues were taken for HE staining.

[0036] Table 1

[0037]

[0038] The results are as follows Figure 3 As shown, the mice in the Model group showed obvious psoriasis-like lesions such as back skin erythema, skin thickening, and desquamation, while the symptoms of the mice in the FN-1501 group were significantly alleviated.

[0039] like Figure 4 As shown in the figure, under the induction of imiquimod, the PASI scores of mice in the Model group gradually increased and skin damage worsened. FN-1501 significantly inhibited the increase in PASI scores of psoriasis mice induced by imiquimod. The results are expressed as Mean ± SEM, and one-way ANOVA was used for variance analysis. Compared with the Control group, # P<0.05, ## P<0.01, ### P<0.001; compared with the Model group, * P<0.05, ** P<0.01, *** P<0.001.

[0040] like Figure 5 As shown, HE staining results showed that compared with the Control group, the proliferation of keratinocytes in the Model group mice increased, and the infiltration of inflammatory cells in the dermis increased. The epidermal hyperplasia and inflammatory cell infiltration in the FN-1501 group were alleviated. Bar = 50μm.

[0041] Example 3: Pharmacological study of FN-1501 in a mouse rheumatoid arthritis model

[0042] Male DBA / 1 mice, 6-8 weeks old, weighing 18-22g, were fed with free water and food, with a 12h light-dark cycle per day and an ambient temperature of 25±2℃. Adaptive feeding was performed for 2 weeks before the start of the study. The mice were randomly divided into 3 groups: normal control group (Control), model control group (Model), and FN-1501 group (FN-1501). Collagen-induced mouse arthritis model (CIA) was established by subcutaneous injection of type II collagen and an equal amount of Freund's complete adjuvant. On the 21st day, type II collagen and an equal amount of Freund's complete adjuvant were injected again to enhance the immune effect. Clinical scoring of the ankle joints of mice was performed from the 21st day, and the specific scoring criteria were as shown in Table 2. FN-1501 (10 mg / kg, iv) was given from the 26th day after modeling, twice a week for 3 weeks. The ankle joint tissue was taken for HE staining on the 47th day after modeling.

[0043] Table 2

[0044]

[0045] The results are as follows Figure 6As shown, FN-1501 significantly reduced the ankle clinical score index of CIA mice. The results are expressed as Mean ± SEM, and one-way ANOVA was used for variance analysis. Compared with the Control group, ## P<0.01, ### P<0.001; compared with the Model group, ** P<0.01.

[0046] like Figure 7 As shown, the joint surface of the mice in the Control group was smooth, and no pathological changes were found in the synovial tissue and cartilage tissue; the synovial tissue of the mice in the Model group showed a large number of inflammatory cell infiltration, hyperplasia and pannus, and local cartilage and bone damage in the joints; compared with the Model group, FN-1501 significantly inhibited synovial inflammation and fibrous tissue hyperplasia, cartilage and bone damage in CIA mice. Bar = 50μm.

Claims

1. Use of FN-1501 in the preparation of a drug for treating and / or preventing autoimmune diseases, wherein the autoimmune diseases include psoriasis and rheumatoid arthritis; the CAS number of the FN-1501 is: 1429515-59-2.

2. Use of a pharmaceutical composition in preparing a drug for treating and / or preventing an autoimmune disease; the pharmaceutical composition is prepared by using FN-1501 or a pharmaceutically acceptable salt or crystal form thereof as an active ingredient and adding a pharmaceutically acceptable carrier; the autoimmune disease includes psoriasis and rheumatoid arthritis; the CAS number of FN-1501 is: 1429515-59-2.

3. The use according to claim 2, characterized in that: The pharmaceutically acceptable salt is a salt formed by FN-1501 and one or more of the following acids: hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, nitrous acid, citric acid, tartaric acid, fumaric acid, methanesulfonic acid, and maleic acid.

4. The use according to claim 2, characterized in that: The pharmaceutically acceptable crystal form is the crystal form corresponding to the free base.

5. The use according to claim 2, characterized in that: The dosage forms of the pharmaceutical composition include tablets, capsules, granules, powders, powder injections, pills, emulsions, suspensions, solutions, sprays, and patches.

6. The use according to claim 2, characterized in that: The pharmaceutically acceptable carrier includes a diluent, a binder, a disintegrant, a glidant, a lubricant, a flavoring agent, an inclusion material, and an adsorption material.

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