Application of C35H39N3O3 small molecule compound or derivative thereof in preparation of medicine for treating autoimmune diseases

Abstract

The invention discloses an application of a small molecule compound with a molecular formula of C35H39N3O3 or a derivative thereof in preparation of a medicine for treating autoimmune diseases. The structural formula of the small molecule compound with the molecular formula of C35H39N3O3 is # imgabs0. The compound has a remarkable effect on treating the autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis and sicca syndrome.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine and relates to a C 35 H 39 Application of N3O3 small molecule compounds or their derivatives in the preparation of drugs for treating autoimmune diseases. Background Art

[0002] Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by loss of self-tolerance, formation of nuclear autoantigens and immune complexes, and multi-organ inflammation. The clinical manifestations of SLE are diverse and can affect one or more organs, including the skin, kidneys, joints, and nervous system. The etiology of SLE is still unclear, but it may be related to many factors, including genetics, environment, immune system, sex hormones, and epigenetics. It is well known that autoreactive CD4+ T cells are essential for the development of SLE, and impaired metabolic homeostasis is the basis of pathogenic CD4+ T cells in SLE. However, the key metabolic nodes that promote the development of autoreactive CD4+ T cells in SLE remain poorly understood. The standardized mortality rate of SLE is estimated to be 2.5% to 5.9%.

[0003] There are many treatment methods currently, mainly immunomodulation and immunosuppression, but they are also limited by drug toxicity and are not effective in improving patient survival rate and prognosis. These are urgent issues that need to be addressed in the treatment of SLE.

[0004] The molecular formula is C 35 H 39 The small molecule compound of N3O3 has the structural formula: Studies have shown that the compound has an inhibitory effect on breast cancer cell growth. At different concentrations, the compound has the effect of promoting oxidative stress levels and inducing autophagy in breast cancer cells, thereby achieving the effect of inhibiting cancer cell proliferation. The present invention is completely different from this. Although the increased differentiation of TFH is not the direct cause of body damage, the present invention explores the use of the compound and its derivatives in the treatment of autoimmune diseases and obtains significant results. Summary of the invention

[0005] The purpose of the present invention is to provide a medicine for treating autoimmune diseases in view of the shortcomings of the prior art.

[0006] In order to solve the above technical problems, the technical solution adopted by the present invention is as follows:

[0007] A molecular formula is C 35 H 39 Application of a small molecule compound of N3O3 or a derivative thereof in the preparation of a drug for treating autoimmune diseases, wherein the C 35 H 39The structural formula of the small molecule compound of N3O3 is:

[0008]

[0009] Preferably, the molecular formula is C 35 H 39 The structural formula of the derivative of the small molecule compound of N3O3 is:

[0010]

[0011] Any one of .

[0012] The autoimmune disease can be systemic lupus erythematosus, rheumatoid arthritis, syndrome, systemic sclerosis, pemphigus, inflammatory bowel disease, myasthenia gravis, Graves' disease, Hashimoto's thyroiditis, primary antiphospholipid antibody syndrome, multiple sclerosis, idiopathic optic neuritis, and neuromyelitis optica spectrum disorder.

[0013] Preferably, the autoimmune disease may also be a TFH cell-related autoimmune disease. Most preferably, the TFH cell-related autoimmune disease is systemic lupus erythematosus.

[0014] Preferably, the above-mentioned drug dosage form includes any one of injection, infusion, powder, tablet, capsule or ointment.

[0015] The molecular formula is C 35 H 39 Small molecule compounds of N3O3 or their derivatives can be used in the preparation of autoimmune disease detection, diagnosis or prognosis products, especially in the preparation of TFH cell-related autoimmune disease detection, diagnosis or prognosis products.

[0016] The present invention uses a dosage of about 50 mg / kg / day for intervention therapy. After drug intervention, the effect is significant, the lupus phenotype is inhibited, and the disease progression of mice is effectively alleviated. The molecular formula is C 35 H 39 Small molecule compounds of N3O3 can inhibit lupus disease by inhibiting TFH differentiation and inhibiting the proliferation of B cells.

[0017] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0018] Excessive TFH cell response drives the production of pathogenic autoantibodies in autoimmune diseases. 35 H 39 Administration of small molecule compounds of N3O3 to lupus model mice can inhibit lupus disease symptoms and inhibit TFH cell differentiation, which can effectively alleviate lupus symptoms. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The results of using flow cytometry to determine the proportion of immune cells in MRL / lpr lupus model mice, using hematoxylin-eosin staining to observe the kidney and skin tissue structure, and using immunofluorescence analysis to determine the expression levels of C3 and IgG in kidney tissue.

[0020] In the figure, A shows the results of weight changes of mice in different groups after ZR001 was used to treat MRL / lpr lupus model mice in an embodiment of the present invention.

[0021] B is the result of reducing the uric acid level in lupus mice after using ZR001 to treat MRL / lpr lupus model mice in an example of the present invention.

[0022] C is the result of reducing the anti-ds-DNA antibody level of lupus mice after using ZR001 to treat MRL / lpr lupus model mice in an example of the present invention.

[0023] D is the result of reducing the antinuclear antibody level of lupus mice after using ZR001 to treat MRL / lpr lupus model mice in an example of the present invention.

[0024] E is the result of the change in the survival rate of mice after ZR001 was used to treat MRL / lpr lupus model mice in an example of the present invention.

[0025] FG is the result of using ZR001 to treat MRL / lpr lupus model mice in an example of the present invention, which inhibited the abnormal enlargement of the spleen and the abnormal increase of spleen cells in lupus mice.

[0026] HI is the result of using ZR001 to treat MRL / lpr lupus model mice in an example of the present invention, which inhibited the abnormal enlargement of lymph nodes and the abnormal increase of lymph node cells in lupus mice.

[0027] J is the result of reducing the kidney organ coefficient after using ZR001 to treat MRL / lpr lupus model mice in the example.

[0028] K is the result of reducing the incidence of skin lesions after using ZR001 to treat MRL / lpr lupus model mice in an example of the present invention.

[0029] L is the result of alleviating kidney damage after using ZR001 to treat MRL / lpr lupus model mice in an example of the present invention.

[0030] M is the result of inhibiting skin thickening after using ZR001 to treat MRL / lpr lupus model mice in an example of the present invention.

[0031] NO is the result of inhibiting the expression of C3 and IgG in kidney tissue after ZR001 was used to treat MRL / lpr lupus model mice in the examples of the present invention.

[0032] PQ is the result of reducing the number of CD4+T cells after using ZR001 to treat MRL / lpr lupus model mice in an example of the present invention.

[0033] R is the result of inhibiting CD4+T cell proliferation after using ZR001 to treat MRL / lpr lupus model mice in an example of the present invention.

[0034] S is the result of reducing the proportion of Tfh cells after using ZR001 to treat MRL / lpr lupus model mice in an example of the present invention. DETAILED DESCRIPTION

[0035] The present invention is further described below in conjunction with specific examples, which are carried out under the preferred conditions of the present invention. The experimental methods described in the following examples are conventional methods unless otherwise specified, and are carried out according to the techniques or conditions described in the literature in the art or according to the product instructions. The materials, reagents, etc. described in the following examples, unless otherwise specified, can all be obtained from commercial sources.

[0036] In the following examples, the flow cytometry, hematoxylin-eosin staining, immunofluorescence analysis and other experimental methods are all conventional experimental methods well known to those skilled in the art. If the experimental method does not specify specific conditions, it is usually performed under conventional conditions.

[0037] The molecular formula used in the following examples is C 35 H 39 The small molecule compound of N3O3 (abbreviated as: ZR001) was prepared according to the method of Example 1 in CN116410152 A patent.

[0038] Example 1: ZR001 alleviates disease progression in MRL / lpr lupus model mice

[0039] MRL / lpr mice of approximately 11 weeks of age were randomly divided into five groups, including a model group, a cyclophosphamide group, a ZR001 low-dose group, a ZR001 medium-dose group, and a ZR001 high-dose group, with 10 mice in each group. The ZR001 low, medium, and high groups were administered 25 mg / kg / day, 50 mg / kg / day, and 100 mg / kg / day, respectively, with a positive drug dose of 15 mg / kg, twice a week. The mice were weighed weekly, the urine protein concentration of the mice was determined using a protein test strip, the anti-dsDNA and antinuclear antibody levels were determined using a kit, and the death of the mice was observed. After 14 weeks of administration, the body weight was weighed at the end of the total experiment, and the urine protein concentration of the mice was determined using a urine protein test strip. After anesthesia, blood, spleen, and lymph nodes were then collected for further analysis. Flow cytometry was used to determine the proportion of immune cells, hematoxylin-eosin staining was used to observe the kidney and skin tissue structure, and immunofluorescence analysis was used to determine the expression levels of C3 and IgG in kidney tissue. The results are shown in the figure. Figure 1 AS shown.

[0040] Figure 1 A is the result of the weight changes of mice in different groups after ZR001 was used to treat MRL / lpr lupus model mice. After using ZR001, there was no significant difference in the weight of mice in each group, indicating that ZR001 had no obvious toxicity at the therapeutic dose.

[0041] Figure 1 B is the result of reducing the uric acid level of lupus mice after using ZR001 to treat MRL / lpr lupus model mice in the example. The high, medium and low dose treatment groups can significantly alleviate the increase of uric acid in lupus mice. After four weeks, the uric acid level of the treatment group showed a significant statistical difference compared with the model group, indicating that the abnormal increase of uric acid in lupus mice was significantly inhibited. This result shows that the administration of ZR001 can effectively inhibit the increase of uric acid and alleviate the progression of lupus disease.

[0042] Figure 1 C is the result of reducing the anti-ds-DNA antibody level of lupus mice after using ZR001 to treat MRL / lpr lupus model mice in the example. The high, medium and low dose treatment groups can significantly alleviate the increase in anti-ds-DNA antibody levels in lupus mice. In the last two weeks, the uric acid level in the treatment group showed a significant statistical difference compared with the model group, indicating that the abnormal increase in anti-ds-DNA antibodies in lupus mice was significantly inhibited. This result shows that the administration of ZR001 can effectively inhibit the increase of anti-ds-DNA antibodies and alleviate the progression of lupus disease.

[0043] Figure 1D is the result of reducing the antinuclear antibody level of lupus mice after using ZR001 to treat MRL / lpr lupus model mice in the example. In the last week, the uric acid level in the treatment group showed a significant statistical difference compared with the model group, indicating that the abnormal increase of antinuclear antibodies in lupus mice was significantly inhibited. This result shows that the administration of ZR001 can effectively inhibit the increase of antinuclear antibodies and alleviate the progression of lupus disease.

[0044] Figure 1 E is the result of the change in the survival rate of mice after ZR001 was used to treat MRL / lpr lupus model mice in the example. The survival rate of mice in the high-dose group was higher than that in the model group, indicating that ZR001 can alleviate the progression of lupus disease and reduce the mortality rate.

[0045] Figure 1 FG is the result of using ZR001 to treat MRL / lpr lupus model mice in the example, which inhibited the abnormal enlargement of the spleen and abnormal increase of spleen cells in lupus mice. The spleen weight and total spleen cell count of mice in the high-dose group were significantly different from those in the model group, indicating that the high-dose group significantly inhibited abnormal enlargement of the spleen and abnormal cell proliferation, which shows that the administration of ZR001 can significantly inhibit the progression of lupus disease.

[0046] Figure 1 HI is the result of inhibiting the abnormal enlargement of lymph nodes and abnormal increase of lymph node cells in lupus mice after ZR001 was used to treat MRL / lpr lupus model mice in the example. The spleen weight and total spleen cell count of mice in the high-dose group were significantly different from those in the model group, indicating that the high-dose group significantly inhibited abnormal enlargement of spleen and abnormal cell proliferation, which shows that administration of ZR001 can significantly inhibit the progression of lupus disease.

[0047] Figure 1 J is the result of reducing the kidney organ coefficient after using ZR001 to treat MRL / lpr lupus model mice in the example. Compared with the model group, the high-dose treatment group significantly inhibited the abnormal increase in kidney mass, indicating that the use of ZR001 treatment can effectively alleviate kidney damage caused by lupus.

[0048] Figure 1 L is the result of alleviating kidney damage after using ZR001 to treat MRL / lpr lupus model mice in the example. Compared with the model group, the glomeruli in the treatment group were significantly reduced. The results show that the use of ZR001 can effectively alleviate kidney damage.

[0049] Figure 1K is the result of reducing the incidence of skin lesions after using ZR001 to treat MRL / lpr lupus model mice in the example. Compared with the model group, the incidence of skin lesions in the treatment group was reduced from 36.4% to 9.1%. The results show that treatment with ZR001 can effectively inhibit the occurrence of skin lesions caused by lupus.

[0050] Figure 1 M is the result of inhibiting skin thickening after using ZR001 to treat MRL / lpr lupus model mice in the example. Compared with the model group, the skin thickening in the treatment group was significantly reduced. The results show that the use of ZR001 can effectively alleviate skin lesions.

[0051] Figure 1 NO is the result of inhibiting the expression of C3 and IgG in kidney tissue after ZR001 was used to treat MRL / lpr lupus model mice in the example. Compared with the model group, the fluorescence signals of C3 and IgG in the treatment group were significantly weakened, indicating that renal inflammation was significantly alleviated after treatment with ZR001, and ZR001 can effectively alleviate kidney damage caused by lupus.

[0052] Figure 1 PQ is the reduction of CD4 in MRL / lpr lupus model mice treated with ZR001 in the example. + Results of T cell counts: Compared with the model group, the ratio of CD4+T cells to the total number of lymph node cells in the treatment group did not change significantly, and the total number of CD4+T cells decreased significantly, indicating that ZR001 treatment can significantly inhibit the proliferation of CD4+T cells.

[0053] Figure 1 R is the inhibition of CD4 in the MRL / lpr lupus model mice treated with ZR001 in the example. + Compared with the model group, the CD4 + The fluorescence intensity of T ki67 decreased significantly, indicating that ZR001 treatment could significantly inhibit CD4 + Proliferation of T cells.

[0054] Figure 1 S is the result of using ZR001 to treat MRL / lpr lupus model mice in the example to reduce the proportion of Tfh cells. Compared with the model group, the proportion of Tfh cells in the treatment group accounted for CD4 + The proportion of T cells was significantly reduced, indicating that the use of ZR001 to treat lupus mice can inhibit Tfh differentiation and thus treat lupus.

[0055] The above data confirm that ZR001 plays an important role in Tfh cell differentiation and Tfh cell-related autoimmune diseases.

Claims

1. A compound with the molecular formula C 35 H 39 Application of a small molecule compound of N3O3 or a derivative thereof in the preparation of a drug for treating autoimmune diseases, wherein the C 35 H 39 The structural formula of the small molecule compound of N3O3 is:

2. The use according to claim 1, characterized in that The molecular formula is C 35 H 39 The structural formula of the derivative of the small molecule compound of N3O3 is: Any one of .

3. The use according to claim 1, characterized in that The autoimmune disease is a TFH cell-related autoimmune disease.

4. The use according to claim 1, characterized in that The autoimmune diseases are systemic lupus erythematosus, rheumatoid arthritis, syndrome, systemic sclerosis, pemphigus, inflammatory bowel disease, myasthenia gravis, Graves' disease, Hashimoto's thyroiditis, primary antiphospholipid antibody syndrome, multiple sclerosis, idiopathic optic neuritis, and neuromyelitis optica spectrum disorder.

5. The use according to claim 4, characterized in that: The autoimmune disease is systemic lupus erythematosus.

6. The use according to claim 1, characterized in that The drug dosage form includes any one of injection, infusion, powder, tablet, capsule or ointment.

7. Molecular formula is C 35 H 39 Application of small molecule compounds of N3O3 or their derivatives in the preparation of products for the detection, diagnosis or prognosis of autoimmune diseases.

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