Compounds and their use in treating autoimmune diseases

Abstract

The present invention relates to: a novel compound and its use in treating autoimmune diseases. The pharmaceutical composition for treating or preventing autoimmune diseases comprising the novel compound of the present invention can not only control inflammation, but also repair tissue homeostasis by repairing immune balance and damaged tissues, thus having excellent effects in treating and preventing autoimmune diseases, and also having excellent effects in treating and preventing cancer.

Description

Technical Field

[0001] The present invention relates to novel compounds and their use in treating autoimmune diseases. Background Art

[0002] The human body can be protected from pathogens by immune responses. Biological defense mechanisms against foreign microorganisms such as viruses and bacteria are generally divided into innate immunity and specific immunity, which are mediated by cytokines secreted mainly by immune-related cells.

[0003] The immune system is used to protect the body from antigens (i.e., harmful foreign substances). Types of these antigens include bacteria, viruses, toxins, cancer cells, and blood and tissues from other humans or animals. The immune system produces antibodies to destroy these harmful substances. If an autoimmune condition is present, the immune system cannot distinguish between its body organs and harmful antigens, and may destroy normal tissues. Diseases induced by such responses as described above are autoimmune diseases.

[0004] Aryl Hydrocarbon Receptor (AHR) is a ligand-dependent transcription factor belonging to the PER-ARNT-SIM (PAS) superfamily and is mainly expressed in immune cells, epithelial cells, endothelial cells, and stromal cells of barrier tissues. AHR is an environmental sensor and detects not only xenobiotic ligands such as environmental pollutants (e.g., dioxins) but also physiological ligands produced by cells, microorganisms, and foods.

[0005] The inactivated form of AHR forms a complex (AHR chaperone complex) with the Hsp90:XAP2:p23:Src chaperone in the cytoplasm and maintains a structure with high affinity for the ligand. When AHR is activated after ligand binding, the complex moves to the nucleus, and AHR separates from the chaperone complex and binds to the AHR responsive DNA element (xenobiotic response element, XRE) located in the upstream regulatory region of the target gene to regulate the expression of the target gene. Non-toxic immunomodulatory ligands that can activate AHR in vivo can be developed as new therapeutic agents for autoimmune diseases. Summary of the invention

[0006] Problem that the invention aims to solve

[0007] The object of the present invention is to provide novel compounds, stereoisomers or pharmaceutically acceptable salts thereof.

[0008] Furthermore, another object of the present invention is to provide a novel compound useful for preventing and treating autoimmune diseases, or a stereoisomer or a pharmaceutically acceptable salt thereof.

[0009] In addition, another object of the present invention is to provide a pharmaceutical composition for preventing or treating autoimmune diseases, which comprises the novel compound, its stereoisomer or pharmaceutically acceptable salt.

[0010] Solutions for solving problems

[0011] 1. A compound represented by the following formula 1, a stereoisomer or a pharmaceutically acceptable salt thereof:

[0012] [Formula 1]

[0013]

[0014] (where R 1 To R 4 are each independently hydrogen or halogen, R 5 and R 6 are independently hydrogen or C 1- C 5 alkyl,

[0015] A is C 5 -C 12 A monocyclic group or a bicyclic group,

[0016] Each ring of the cyclic group is substituted by 1 to 3 heteroatoms, and

[0017] Cyclic groups are composed of halogen, C 1 -C 5 Alkyl or C 1 -C 5 alkoxy substituted).

[0018] 2. The compound according to 1 above, its stereoisomer or pharmaceutically acceptable salt, wherein A is selected from the group consisting of the following cyclic groups:

[0019]

[0020]

[0021] (where Q 1 To Q 15 are each independently C, N or S, and R 7 To R 30 are independently hydrogen, halogen, C 1 -C3 Alkyl or C 1 -C 3 Alkoxy, and if Q 4 is N, then R 11 does not exist).

[0022] 3. The compound according to 1 above, its stereoisomer or pharmaceutically acceptable salt, wherein A is selected from the group consisting of the following cyclic groups:

[0023]

[0024] (where R 7 To R 30 are independently hydrogen, halogen, C 1 -C 3 Alkyl or C 1 -C 3 alkoxy).

[0025] 4. The compound according to 1 above, its stereoisomer or pharmaceutically acceptable salt, wherein A is selected from the group consisting of the following cyclic groups:

[0026]

[0027] (where R 7 To R 24 are independently hydrogen, halogen, C 1 -C 3 Alkyl or C 1 -C 3 alkoxy).

[0028] 5. The compound according to 1 above, its stereoisomer or pharmaceutically acceptable salt, wherein A is selected from the group consisting of the following cyclic groups:

[0029]

[0030] (where R 9 To R 16 are independently hydrogen, halogen, C 1 -C 3 Alkyl or C 1 -C 3 alkoxy).

[0031] 6. The compound according to 1 above, its stereoisomer or pharmaceutically acceptable salt, wherein R 2 and R 3 Each is independently F, Cl or Br.

[0032] 7. The compound according to 1 above, its stereoisomer or pharmaceutically acceptable salt, wherein the compound is selected from the group consisting of the following compounds:

[0033] N-(5-bromo-6-methylpyridin-2-yl)-2-(1-methyl-1H-indol-3-yl)acetamide,

[0034] N-(5-bromo-6-methylpyridin-2-yl)-2-(1H-indol-3-yl)acetamide;

[0035] N-(5-bromo-6-methylpyridin-2-yl)-2-(5-chloro-1H-indol-3-yl)acetamide;

[0036] N-(Benzo[di]thiazol-2-yl)-2-(1H-indol-3-yl)acetamide;

[0037] N-(Benzo[di]thiazol-2-yl)-2-(5-chloro-1H-indol-3-yl)acetamide;

[0038] N-(5-chloro-6-fluoropyridin-2-yl)-2-(1H-indol-3-yl)acetamide;

[0039] 2-(5-chloro-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)acetamide;

[0040] 2-(1H-Indol-3-yl)-N-(3,4,5-trimethoxyphenyl)acetamide;

[0041] 2-(5-Chloro-1H-indol-3-yl)-N-(3,4,5-trimethoxyphenyl)acetamide;

[0042] N-(3,5-dichlorophenyl)-2-(1H-indol-3-yl)acetamide;

[0043] 2-(5-Chloro-1H-indol-3-yl)-N-(3,5-dichlorophenyl)acetamide;

[0044] N-(5-bromo-6-methylpyridin-2-yl)-2-(5-fluoro-1H-indol-3-yl)acetamide;

[0045] 2-(5-Chloro-1H-indol-3-yl)-N-(pyridin-4-yl)acetamide;

[0046] N-(Benzo[di]thiazol-2-yl)-N-methyl-2-(1-methyl-1H-indol-3-yl)acetamide;

[0047] N-(Benzo[di]thiazol-2-yl)-2-(1H-indol-3-yl)-N-methylacetamide;

[0048] N-(Benzo[di]thiazol-2-yl)-2-(5-chloro-1H-indol-3-yl)-N-methylacetamide;

[0049] N-(5-chloro-6-fluoropyridin-2-yl)-2-(1H-indol-3-yl)-N-methylacetamide;

[0050] 2-(5-chloro-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)-N-methylacetamide;

[0051] N-(Benzo[di]thiazol-2-yl)-2-(5-chloro-1-methyl-1H-indol-3-yl)-N-methylacetamide;

[0052] N-(5-chloro-6-fluoropyridin-2-yl)-N-methyl-2-(1-methyl-1H-indol-3-yl)acetamide;

[0053] 2-(5-chloro-1-methyl-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)-N-methylacetamide;

[0054] 2-(5-chloro-1-methyl-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)acetamide;

[0055] N-(Benzo[di]thiazol-2-yl)-2-(1-methyl-1H-indol-3-yl)acetamide;

[0056] N-(Benzo[di]thiazol-2-yl)-2-(5-fluoro-1H-indol-3-yl)-N-methylacetamide;

[0057] N-(Benzo[di]thiazol-2-yl)-2-(5-chloro-1-methyl-1H-indol-3-yl)acetamide;

[0058] N-(Benzo[di]thiazol-2-yl)-2-(5-fluoro-1H-indol-3-yl)acetamide;

[0059] N-(Benzo[di]thiazol-2-yl)-2-(6-chloro-1H-indol-3-yl)acetamide;

[0060] 2-(5-Chloro-1H-indol-3-yl)-N-(thiazol-2-yl)acetamide;

[0061] 2-(5-chloro-1H-indol-3-yl)-N-(quinolin-2-yl)acetamide; and

[0062] 2-(5-Chloro-1H-indol-3-yl)-N-(4,5,6,7-tetrahydrobenzo[di]thiazol-2-yl)acetamide.

[0063] 8. A pharmaceutical composition comprising the compound according to any one of 1 to 7 above, a stereoisomer or a pharmaceutically acceptable salt thereof.

[0064] 9. The pharmaceutical composition according to 8 above, wherein the composition is used to treat or prevent autoimmune diseases.

[0065] 10. The pharmaceutical composition according to 8 above, wherein the autoimmune disease is any one selected from the group consisting of multiple sclerosis, inflammatory bowel disease, graft-versus-host disease, asthma, atopy, psoriasis, rheumatoid arthritis, systemic lupus erythematosus and type 1 diabetes.

[0066] 11. The pharmaceutical composition according to 8 above, wherein the composition is used for treating or preventing cancer.

[0067] 12. The pharmaceutical composition according to 11 above, wherein the cancer is selected from the group consisting of melanoma, colon cancer, liver cancer, glioma, ovarian cancer, colon cancer, head and neck cancer, bladder cancer, renal cell carcinoma, gastric cancer, breast cancer, metastatic cancer, prostate cancer, gallbladder cancer, pancreatic cancer, blood cancer, skin cancer and lung cancer.

[0068] 13. A method for treating an autoimmune disease, comprising administering the compound according to any one of 1 to 7 above, a stereoisomer or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0069] 14. The method for treating an autoimmune disease according to 13 above, wherein the autoimmune disease is selected from the group consisting of multiple sclerosis, inflammatory bowel disease, graft-versus-host disease, asthma, atopy, psoriasis, rheumatoid arthritis, systemic lupus erythematosus and type 1 diabetes.

[0070] 15. A method for inducing AHR activity, comprising administering the compound according to any one of 1 to 7 above, a stereoisomer or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0071] 16. A method for inhibiting the production of IL-6, comprising administering the compound according to any one of 1 to 7 above, a stereoisomer or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0072] 17. A method for treating cancer, comprising administering the compound according to any one of 1 to 7 above, its stereoisomer or pharmaceutically acceptable salt to a subject in need thereof.

[0073] 18. The method for treating cancer according to 17 above, wherein the cancer is selected from the group consisting of melanoma, colorectal cancer, liver cancer, glioma, ovarian cancer, colorectal cancer, head and neck cancer, bladder cancer, renal cell carcinoma, gastric cancer, breast cancer, metastatic cancer, prostate cancer, gallbladder cancer, pancreatic cancer, blood cancer, skin cancer and lung cancer.

[0074] Effects of the Invention

[0075] The novel compound, stereoisomer or pharmaceutically acceptable salt thereof according to the present invention can induce the activity of AHR as an immunomodulatory transcription factor, thereby achieving the effect of not only controlling inflammation but also repairing immune balance and damaged tissues.

[0076] The novel compound, stereoisomer or pharmaceutically acceptable salt thereof according to the present invention can inhibit the production of IL-6, which is an inflammatory factor, thereby achieving the effect of regulating excessive immune response, especially autoimmune response.

[0077] The novel compound, stereoisomer or pharmaceutically acceptable salt thereof according to the present invention may exhibit an effect of inducing the activity of regulatory T cells (Treg).

[0078] In addition, the novel compound, stereoisomer or pharmaceutically acceptable salt thereof according to the present invention can exhibit the effect of preventing and treating autoimmune diseases by regulating the above inflammatory factors. BRIEF DESCRIPTION OF THE DRAWINGS

[0079] Figure 1 and Figure 2 Measurement of CYP1A1 expression levels is shown to confirm that the compounds of the invention are AHR ligands under cell culture conditions.

[0080] Figure 3 and Figure 4 The results show the measurement of the inhibitory effect of the compounds of the present invention on the production of inflammatory factor IL-6.

[0081] Figure 5 Shown is the measurement of the Foxp3+ regulatory T cell generating effect of the compounds of the invention.

[0082] Figure 6 and Figure 7 The present invention shows the therapeutic effect of the compound of the present invention on inflammatory bowel disease in an animal model of inflammatory bowel disease induced by dextran sodium sulfate (DSS). Specifically, Figure 6 showed that the lower the severity index, the more treatment was completed compared with the control (vehicle), while Figure 7 It was shown that the smaller the weight of the large intestine compared to its length, the higher the treatment effect of inflammatory bowel disease.

[0083] Figure 8and Fig. 9 The compounds of the present invention were shown to inhibit the expression of inflammatory factors (IL-1β, IL-6, IL-17a and TNF-α) and increase the expression of immunoregulatory factors (IL-10 and Foxp3) in an animal model of DSS-induced inflammatory bowel disease.

[0084] Fig.10 The compounds of the present invention are shown to have a mucosal healing effect in an animal model of DSS-induced inflammatory bowel disease using FITC-dextran. Herein, this means that the lower the degree of detection, the higher the mucosal healing effect.

[0085] Fig.11 The compounds of the present invention are shown to be effective in preventing inflammation-induced colorectal cancer in the AOM / DSS colorectal cancer animal model. Here, this means that the fewer the number of tumors per colon, the more effective it is in preventing colorectal cancer.

[0086] Fig.12 The effect of the compounds of the present invention in treating multiple sclerosis in an experimental autoimmune encephalomyelitis (EAE) animal model is shown. Specifically, in order to confirm the therapeutic effect of multiple sclerosis, the severity index is shown as a graph over time. In this article, this means that the lower the severity index is compared to the control, the more treatment is completed.

[0087] Fig.13 and Fig.14 It is shown that the compounds of the present invention Fig.12 It inhibits the expression of inflammatory factors (IFN-γ, IL-17a and IL-1β) and increases the expression of immunoregulatory factors (IL-10 and Foxp3) in the EAE animal model.

[0088] Fig.15 Graph showing the severity index measured to confirm the therapeutic effect of graft-versus-host disease (GVHD) in an animal model with pulmonary GVHD.

[0089] Fig.16 Shown in Fig.15 The expression levels of IL-6, IL-17a and IL-10 factors were measured by the compounds of the present invention in an animal model. DETAILED DESCRIPTION

[0090] Hereinafter, the present invention will be described in detail.

[0091] Unless otherwise defined, all technical terms used in the present invention are used with the same meaning as those generally understood by those skilled in the art in the relevant field of the present invention. In addition, although preferred methods or samples will be described in this specification, similar or equivalent ones are also included in the scope of the present invention.

[0092] The present invention relates to a compound represented by the following formula 1, a stereoisomer or a pharmaceutically acceptable salt thereof:

[0093] [Formula 1]

[0094]

[0095] In the above formula, if a position requires a substituent but no substituent is specified at that position, this means that the hydrogen substituent is omitted, and this shall apply to all formulae in the present invention.

[0096] In the above formula, R 1 To R 4 Each may be independently hydrogen or a halogen, and specifically hydrogen, fluorine or chlorine, but they are not limited thereto.

[0097] In the above formula, R 5 and R 6 can be each independently hydrogen or C 1 -C 5 The alkyl group is specifically hydrogen, methyl or ethyl, and more specifically hydrogen or methyl, but they are not limited thereto.

[0098] In the above formula, A can be C 5 -C 12 The monocyclic group or bicyclic group is specifically cyclopenta-1,3-diene, benzene, cyclohexane, indene, 4,5,6,7-tetrahydroindene, naphthalene, 1,2,3,4-tetrahydronaphthalene, 1,6-dihydropentalene, etc., but is not limited thereto.

[0099] Each ring of the cyclic group may be substituted with 1 to 3 heteroatoms, and, for example, 1 to 3 heteroatoms may each be independently substituted with N, S, O, etc., but they are not limited thereto. The heteroatom means an atom other than carbon or hydrogen.

[0100] In addition, specifically, the sites where the heteroatom can be substituted may include Q in the structures listed below: 1 To Q 15 , but not limited to this.

[0101]

[0102] In the above formula, if Q 4 is N, then in Q 4 No further substitution can occur at this position, so this can be 11 A situation that does not exist.

[0103] The cyclic group can be composed of halogen, C 1 -C5 Alkyl or C 1 -C 5 Alkoxy groups, for example, F, Cl, methyl, ethyl, methoxy, ethoxy, etc., are substituted, but are not limited thereto.

[0104] The cyclic group can be composed of halogen, C 1 -C 5 Alkyl or C 1 -C 5 The alkoxy substituted sites may specifically include R 7 To R 30 , but not limited to this.

[0105] According to an embodiment of the present invention, A may be selected from the following cyclic groups.

[0106]

[0107] (where R 7 To R 30 are independently hydrogen, halogen, C 1 -C 3 Alkyl or C 1 -C 3 alkoxy).

[0108] According to an embodiment of the present invention, A may be specifically selected from the following cyclic groups.

[0109]

[0110] (where R 7 To R 24 are independently hydrogen, halogen, C 1 -C 3 Alkyl or C 1 -C 3 alkoxy).

[0111] Furthermore, according to an embodiment of the present invention, A may be more specifically selected from the following cyclic groups.

[0112]

[0113] (where R 9 To R 16 are independently hydrogen, halogen, C 1 -C 3 Alkyl or C 1 -C 3 alkoxy).

[0114] Table 1 below shows examples of the structure of the compound represented by Formula 1, which is represented by R 1 To R 6 The combination with A is specifically defined.

[0115] [Table 1]

[0116]

[0117]

[0118]

[0119] The present invention relates to a compound selected from the group consisting of the following compounds and stereoisomers or pharmaceutically acceptable salts thereof.

[0120] N-(5-bromo-6-methylpyridin-2-yl)-2-(1-methyl-1H-indol-3-yl)acetamide;

[0121] N-(5-bromo-6-methylpyridin-2-yl)-2-(1H-indol-3-yl)acetamide;

[0122] N-(5-bromo-6-methylpyridin-2-yl)-2-(5-chloro-1H-indol-3-yl)acetamide;

[0123] N-(Benzo[di]thiazol-2-yl)-2-(1H-indol-3-yl)acetamide;

[0124] N-(Benzo[di]thiazol-2-yl)-2-(5-chloro-1H-indol-3-yl)acetamide;

[0125] N-(5-chloro-6-fluoropyridin-2-yl)-2-(1H-indol-3-yl)acetamide;

[0126] 2-(5-chloro-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)acetamide;

[0127] 2-(1H-Indol-3-yl)-N-(3,4,5-trimethoxyphenyl)acetamide;

[0128] 2-(5-Chloro-1H-indol-3-yl)-N-(3,4,5-trimethoxyphenyl)acetamide;

[0129] N-(3,5-dichlorophenyl)-2-(1H-indol-3-yl)acetamide;

[0130] 2-(5-Chloro-1H-indol-3-yl)-N-(3,5-dichlorophenyl)acetamide;

[0131] N-(5-bromo-6-methylpyridin-2-yl)-2-(5-fluoro-1H-indol-3-yl)acetamide;

[0132] 2-(5-Chloro-1H-indol-3-yl)-N-(pyridin-4-yl)acetamide;

[0133] N-(Benzo[di]thiazol-2-yl)-N-methyl-2-(1-methyl-1H-indol-3-yl)acetamide;

[0134] N-(Benzo[di]thiazol-2-yl)-2-(1H-indol-3-yl)-N-methylacetamide;

[0135] N-(Benzo[di]thiazol-2-yl)-2-(5-chloro-1H-indol-3-yl)-N-methylacetamide;

[0136] N-(5-chloro-6-fluoropyridin-2-yl)-2-(1H-indol-3-yl)-N-methylacetamide;

[0137] 2-(5-chloro-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)-N-methylacetamide;

[0138] N-(Benzo[di]thiazol-2-yl)-2-(5-chloro-1-methyl-1H-indol-3-yl)-N-methylacetamide;

[0139] N-(5-chloro-6-fluoropyridin-2-yl)-N-methyl-2-(1-methyl-1H-indol-3-yl)acetamide;

[0140] 2-(5-chloro-1-methyl-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)-N-methylacetamide;

[0141] 2-(5-chloro-1-methyl-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)acetamide;

[0142] N-(Benzo[di]thiazol-2-yl)-2-(1-methyl-1H-indol-3-yl)acetamide;

[0143] N-(Benzo[di]thiazol-2-yl)-2-(5-fluoro-1H-indol-3-yl)-N-methylacetamide;

[0144] N-(Benzo[di]thiazol-2-yl)-2-(5-chloro-1-methyl-1H-indol-3-yl)acetamide;

[0145] N-(Benzo[di]thiazol-2-yl)-2-(5-fluoro-1H-indol-3-yl)acetamide;

[0146] N-(Benzo[di]thiazol-2-yl)-2-(6-chloro-1H-indol-3-yl)acetamide;

[0147] 2-(5-Chloro-1H-indol-3-yl)-N-(thiazol-2-yl)acetamide;

[0148] 2-(5-chloro-1H-indol-3-yl)-N-(quinolin-2-yl)acetamide; and

[0149] 2-(5-Chloro-1H-indol-3-yl)-N-(4,5,6,7-tetrahydrobenzo[di]thiazol-2-yl)acetamide;

[0150] Furthermore, the present invention relates to a pharmaceutical composition comprising the compound, its stereoisomer or a pharmaceutically acceptable salt.

[0151] The pharmaceutical composition can be a pharmaceutical composition for treating or preventing an autoimmune disease. Specifically, the disease can be multiple sclerosis (MS), inflammatory bowel disease (IBD), graft versus host disease (GVHD), asthma, atopy, psoriasis, rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), type 1 diabetes (T1D), Behcet's disease or Sjögren's syndrome. More specifically, the disease can be multiple sclerosis, inflammatory bowel disease, graft versus host disease, asthma, atopy, psoriasis, rheumatoid arthritis, systemic lupus erythematosus, type 1 diabetes, but is not limited thereto.

[0152] In the present invention, "autoimmune disease" may cause damage to cells or tissues through humoral immunity, cellular immunity, or both. That is, autoimmune disease is a disease in which the immune system produces an inappropriate response to self-antigens, thereby inducing autoimmune responses systemically or specifically in specific organs, which may cause chronic inflammation.

[0153] "Multiple sclerosis" broadly refers to an inflammatory disease that causes demyelination and scarring as signs and symptoms, caused by damage and / or consumption of the fatty myelin sheaths around the axons of the brain and spinal cord. Types of multiple sclerosis may include relapsing-remitting multiple sclerosis (RRMS), secondary progressive multiple sclerosis (SPMS), primary progressive multiple sclerosis (PPMS), and progressive recurrent multiple sclerosis (PRMS), but they are not limited thereto.

[0154] “Inflammatory bowel disease” refers to a disease in which abnormal chronic inflammation in the intestine repeatedly improves and relapses, and may correspond to one selected from the group consisting of Crohn's disease, ulcerative colitis, and intestinal Behcet's disease, but is not limited thereto.

[0155] "Graft-versus-host disease" is a disease in which lymphocytes infused during hematopoietic stem cell transplantation attack a host with reduced immune function to cause symptoms such as fever, rash, and abnormal liver function, and may affect the skin, lungs, intestines, liver, etc., but is not limited thereto.

[0156] "Asthma" refers to a disease in which symptoms such as coughing and breathing difficulties recur due to inflammation of the bronchial tubes when exposed to a specific pathogen, and may be caused by infection, smoking, allergens, etc., but is not limited thereto.

[0157] “Atopy” means atopic dermatitis, and is a representative allergic disease that presents symptoms such as itching and dry skin as a chronic recurring inflammatory skin disease.

[0158] "Psoriasis" refers to an inflammatory disease that occurs in the skin or joints due to abnormalities in the immune system and causes problems such as the appearance of ugly, increased keratin or erythematous plaques accompanied by pain. Psoriasis may include any one or more diseases selected from psoriatic arthritis, guttate psoriasis, pustular psoriasis, red skin psoriasis, scalp psoriasis, nail psoriasis, and enthesitis.

[0159] "Rheumatoid arthritis" refers to a systemic autoimmune disease characterized by chronic inflammation of the joints.

[0160] "Systemic lupus erythematosus" is also called "lupus," and refers to a systemic disease that is a chronic inflammatory autoimmune disease that attacks various organs of the body, such as connective tissue, skin, joints, blood, and kidneys. The exact cause is unknown, but according to previous studies, genetic factors are known to be involved in the development of the disease. To help diagnose lupus, the American College of Rheumatology (ACR) publishes 11 symptoms, signs, and test results to help distinguish the disease from other diseases. According to published research, if more than 4 of the 11 symptoms are present, lupus can be diagnosed.

[0161] "Type 1 diabetes" is an immune-mediated disease in which the insulin-secreting beta cells are destroyed by an autoimmune response. The causes of the disease can include a number of genetic and environmental factors that specifically target the insulin-secreting beta cells. The disease may be accompanied by progressive inflammatory infiltration of the pancreatic islets by immune cells.

[0162] In the present invention, in addition to the active ingredient of the compound of the present invention, pharmaceutically suitable and physiologically acceptable additives can also be used to prepare a pharmaceutical composition. The composition can be administered to a mammal. As the above-mentioned additives, for example, excipients, disintegrants, sweeteners, binders, coating agents, leavening agents, lubricants, glidants or flavoring agents can be used.

[0163] Furthermore, the pharmaceutical composition of the present invention may be preferably formulated as a pharmaceutical composition comprising at least one pharmaceutically acceptable carrier in addition to the above-mentioned pharmaceutically effective amount of the active ingredient for administration.

[0164] "Pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio suitable for drug therapy, and can be based on the type and severity of the patient's disease, drug activity, drug sensitivity, administration time, route of administration and excretion rate, treatment duration, factors including drugs used simultaneously, and other factors known to the medical field to determine the effective dose level. Pharmaceutical compositions according to the present invention can be administered as a single therapeutic agent or in combination with other therapeutic agents. In addition, compositions can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered in single doses or multiple doses. Taking all of the above factors into account, it is important to give the minimum amount that can achieve maximum effect without side effects, and such an amount can be easily determined by those skilled in the art.

[0165] Particularly, the effective amount of the pharmaceutical composition according to the present invention can depend on the patient's age, sex, patient's condition and / or body weight, absorption of active ingredient in vivo, inactivation rate and excretion rate, disease type and the medicine to be used in combination and change.Usually, 0.001 to 150mg, preferably 0.01 to 100mg per 1kg body weight can be administered every day or every other day, or can be divided into 1 to 3 times a day.However, dosage can increase or decrease depending on route of administration, severity of obesity, sex, body weight, age, etc., therefore, will not limit the scope of the present invention in any way.

[0166] Furthermore, "pharmaceutically acceptable" refers to compositions that are physiologically acceptable and do not generally cause allergic reactions or the like, such as gastrointestinal disturbances and dizziness, when administered to humans.

[0167] The example of carrier, excipient and diluent can comprise lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil. In addition, filler, anti-aggregating agent, lubricant, wetting agent, flavoring agent, emulsifier and preservative can be additionally included.

[0168] In addition, the compositions of the present invention can be formulated using any method known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a subject, including a human, who needs to be treated with the pharmaceutical composition of the present invention. The preparation can be a powder, granules, tablet, emulsion, syrup, aerosol, soft or hard gelatin capsule, sterile injection, sterile powder.

[0169] The present invention may relate to a method for treating an autoimmune disease, comprising administering the compound, a stereoisomer or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0170] Furthermore, the present invention may relate to a method for inducing the activity of AHR, which comprises administering the compound, a stereoisomer or a pharmaceutically acceptable salt thereof.

[0171] Specifically, the compounds of the present invention can target the aryl hydrocarbon receptor (AHR) as the immunomodulatory transcription factor of the present invention, and can be used as an agent for inducing AHR activity, thereby controlling inflammation, regulating immune balance and repairing damaged tissues. Therefore, the compounds can be used for therapeutic purposes, but are not limited to this. Existing ligands are toxic, have low affinity and structural stability and high target non-specificity, which brings about the problem that these compounds are not suitable for development into pharmaceutical compositions. On the other hand, when AHR activity is induced by a compound of the present invention having "drug-like properties", it can be effectively used to treat and prevent autoimmune diseases.

[0172] The present invention may relate to a method for inhibiting the production of IL-6, which comprises administering the compound, a stereoisomer or a pharmaceutically acceptable salt thereof.

[0173] Specifically, since it is known that IL-6 (an inflammatory factor) can cause autoimmune diseases, the compounds of the present invention can be used to treat autoimmune diseases by inhibiting the mechanism of IL-6 production. In fact, there are many known therapeutic agents for autoimmune diseases that inhibit the inhibition of IL-6, as well as related papers. According to the following experimental data, the compounds of the present invention are also confirmed to inhibit the production of IL-6 and are therefore expected to have the effect of reducing autoimmune responses, thus, the compositions of the present invention can be used to treat and prevent autoimmune diseases.

[0174] Furthermore, the present invention relates to a composition for preventing or treating cancer, which comprises the compound, a stereoisomer or a pharmaceutically acceptable salt thereof.

[0175] In the present invention, "cancer" refers to the uncontrolled abnormal growth of the host's own cells, which invades the surrounding tissues of the initial abnormal cell growth site in the host and potential tissues away from these sites. In addition, it can include carcinoma as a cancer of epithelial tissue (e.g., skin, squamous cells); sarcoma as a cancer of connective tissue (e.g., bone, cartilage, fat, muscle, blood vessels, etc.); leukemia as a cancer of hematopoietic tissue (e.g., bone marrow tissue); lymphoma and myeloma as cancer of immune cells; cancer of the central nervous system, including cancer in brain and spinal cord tissue.

[0176] Specifically, the cancer can be selected from the group consisting of melanoma, colorectal cancer, liver cancer, glioma, ovarian cancer, colon cancer, head and neck cancer, bladder cancer, renal cell carcinoma, gastric cancer, breast cancer, metastatic cancer, prostate cancer, gallbladder cancer, pancreatic cancer, blood cancer, skin cancer and lung cancer, but is not limited thereto.

[0177] The present invention relates to a method for treating cancer, which comprises administering the compound, its stereoisomer or pharmaceutically acceptable salt to a subject in need thereof.

[0178] The treatment method may include administering the compound, a stereoisomer or a pharmaceutically acceptable salt thereof to a patient diagnosed with cancer at any stage of chemotherapy and is not limited to a specific stage.

[0179] Furthermore, the compound, its stereoisomer or pharmaceutically acceptable salt may be administered in the form of the aforementioned pharmaceutical composition, but is not limited thereto.

[0180] The compound represented by Formula 1 according to the present invention can be prepared by any method known in various literatures. In the following preparation examples, the synthesis methods of some compounds listed in Table 1 are briefly described, however, they are not limited thereto.

[0181] Hereinafter, the present invention will be described in detail through Preparation Examples and Examples of the present invention.

[0182] Preparation Example

[0183] 1. Synthesis of N-(5-bromo-6-methylpyridin-2-yl)-2-(5-chloro-1H-indol-3-yl)acetamide (Compound 8)

[0184] [Process 1]

[0185]

[0186] 2-(5-Chloro-1H-indol-3-yl)acetic acid (1.00 g, 4.77 mmol) was stirred in CH 2 Cl 2 (30mL) solution, 5-bromo-6-methylpyridin-2-amine (892mg, 4.77mmol), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide hexafluorophosphate (HATU, 2.18g, 5.72mmol) and trimethylamine (1.33mL, 9.54mmol) were added dropwise in sequence. The reaction mixture was stirred at room temperature for 3 days, and distilled water (10mL) was added to the mixture to terminate the reaction. The layers were separated, and the organic layer was washed with distilled water, washed with anhydrous Na 2 SO 4 After the filtrate was concentrated under reduced pressure, the concentrate was purified by column chromatography (SiO 2 , hexane:EtOAc=4:1-2:1) to give a light grey compound (970 mg, 54%).

[0187] 1 H NMR (CDCl 3 ,400MHz): δ8.67(br s,1H),8.00(d,J=8.0Hz,1H),7.97(br s,1H),7.55(d,J=4.0Hz,1H),7.23(d,J=8.0Hz,1H),7.14(m,2H),3.84(s,2H),2.45(s,3H).

[0188] 2. Synthesis of N-(Benzo[di]thiazol-2-yl)-2-(5-chloro-1H-indol-3-yl)acetamide (Compound 40)

[0189] [Process 2]

[0190]

[0191] The title compound (1.31 g, 80%) was obtained as a white color by the same experimental procedure as in Preparation Example 1, except that the amine in Preparation Example 1 was changed to benzo[di]thiazol-2-amine.

[0192] 1 H NMR(DMSO-d6,400MHz): δ12.58(br s,1H),11.20(br s,1H),7.95(m,1H),7.74(d,J=8.0Hz,1H),7.68(d,J=4.0Hz,1H),7.43(ddd,J=8.0,8.0,2.0Hz ,1H),7.39(m,2H),7.29(ddd,J=8.0,8.0,2.0Hz,1H),7.09(dd,J=8.0,4.0Hz,1H),3.91(s,2H).

[0193] 3. Synthesis of 2-(5-chloro-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)acetamide (Compound 26)

[0194] [Process 3]

[0195]

[0196] The pale yellow title compound (560 mg, 35%) was obtained by the same experimental procedure as in Preparation Example 1, except that the amine in Preparation Example 1 was changed to 5-chloro-6-fluoropyridin-2-amine.

[0197] 1 H NMR (CDCl 3 ,400MHz):δ8.38(br s,1H),8.14(dd,J=8.0,2.0Hz,1H),7.87(brs,1H),7.77(dd,J=8.0,8.0Hz,1H),7 .54(dd,J=4.0,2.0Hz,1H),7.33(dd,J=8.0,0.8Hz,1H),7.21(m,2H),3.87(s,2H).

[0198] 4. Synthesis of N-(5-bromo-6-methylpyridin-2-yl)-2-(5-fluoro-1H-indol-3-yl)acetamide (Compound 2)

[0199] [Process 4]

[0200]

[0201] The title compound (107 mg, 44%) was obtained as a light brown color by the same experimental procedure as in Preparation Example 1, except that the acetic acid in Preparation Example 1 was changed to 2-(5-chloro-1H-indol-3-yl)acetic acid.

[0202] 1 H NMR (CDCl 3 ,400MHz): δ9.09(br s,1H),8.20(br s,1H),8.02(d,J=8.0Hz,1H),7.76(d,J=8.0Hz,1H),7.20(dd,J=8.0,4.0Hz,1H),7.13 (dd,J=8.0,4.0Hz,1H),6.98(d,J=4.0Hz,1H),6.89(m,1H),3.83(s,2H),2.44(s,3H).

[0203] 5. Synthesis of N-(Benzo[di]thiazol-2-yl)-N-methyl-2-(1-methyl-1H-indol-3-yl)acetamide (Compound 45)

[0204] While a solution of N-(benzo[di]thiazol-2-yl)-2-(1H-indol-3-yl)acetamide (70.0 mg, 0.228 mmol) in DMF (1 mL) was stirred at room temperature under an argon atmosphere, t-BuOK (74.0 mg, 0.456 mmol) was added dropwise and stirred for 5 minutes. MeI (28.4 μL, 0.456 mmol) was added dropwise to the mixture and stirred for 30 minutes. Distilled water (1 mL) was added to the mixture to terminate the reaction. The layers were separated, and the organic layer was washed with distilled water, filtered, and washed with anhydrous MgSO 4 After the filtrate was concentrated under reduced pressure, the concentrate was purified by column chromatography (SiO 2 , hexane:EtOAc=4:1) to give the title compound (39.0 mg, 51%) as a white color.

[0205] 1 H NMR (CDCl 3 ,500MHz): δ7.83(d,J=8.1Hz,1H),7.80(d,J=7.9Hz,1H),7.64(d,J=8.0Hz,1H),7.43(td,J= 7.8,1.0Hz,1H),7.29(m,3H),7.16(t,J=7.5Hz,1H),4.17(s,2H),3.85(s,3H),3.77(s,3H).

[0206] 13 C NMR (CDCl 3,125MHz): δ171.87,160.26,148.19,137.08,133.59,127.73,127.56,126.01,123. 92,122.20,121.36,121.18,119.61,118.76,109.60,105.76,35.73,32.93,32.87.

[0207] 6. Synthesis of N-(5-chloro-6-fluoropyridin-2-yl)-2-(1H-indol-3-yl)-N-methylacetamide (Compound 35)

[0208] (1) Stage 1: Synthesis of 2-(1H-indol-3-yl)acetyl chloride

[0209] Indole-3-acetic acid (39.3 mg, 0.224 mmol) was stirred in CH at 0 °C under argon atmosphere. 2 Cl 2 (1.5 mL), oxalyl chloride (96.0 μL, 1.12 mmol) and DMF (1 drop) were added dropwise in sequence. The reaction mixture was stirred for 1 hour. The mixture was concentrated under reduced pressure, dried in vacuo and used in the next reaction without further purification.

[0210] (2) Stage 2: Synthesis of N-(5-chloro-6-fluoropyridin-2-yl)-2-(1H-indol-3-yl)-N-methylacetamide

[0211] While stirring a solution of 5-chloro-6-fluoro-N-methylpyridin-2-amine (30.0 mg, 0.187 mmol) in THF (1 mL) at 0°C under argon atmosphere, n-BuLi (116 μL, 0.187 mmol) was added dropwise. The reaction mixture was stirred for 1 hour. 2-(1H-indol-3-yl)acetyl chloride from stage 1 was added in CH 2 Cl 2 The solution in (0.5 mL) was added dropwise to the mixture. After the mixture was stirred for 5 minutes, distilled water (1 mL) was added to terminate the reaction. The layers were separated, and the organic layer was washed with distilled water, washed with anhydrous MgSO 4 After the filtrate was concentrated under reduced pressure, the concentrate was purified by column chromatography (SiO 2 , Hexane:EtOAc:CH 2 Cl 2 =3:3:1) to give the brown title compound (19.0 mg, 27%).

[0212] 1 H NMR (CDCl 3,500MHz): δ8.15(s,1H),7.69(t,J=8.7Hz,1H),7.53(d,J=7.9Hz,1H),7.34(s,1H),7.31(s ,1H),7.19(t,J=7.5Hz,1H),7.11(t,J=7.4Hz,1H),7.04(s,1H),3.98(s,2H),3.43(s,3H).

[0213] 13 C NMR (CDCl 3 ,125MHz): δ172.17,159.28,157.85,142.29,141.91,141.90,136.20,127.17,122.94,1 22.44,119.86,118.77,118.01,117.97,117.77,117.73,111.36,108.76,35.60,29.83.

[0214] 7. Synthesis of 2-(5-chloro-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)-N-methylacetamide (Compound 32)

[0215] The title compound (11.6 mg, 15%) in brown color was obtained by the same experimental procedure as in Preparation Example 6, except that the acetic acid in Preparation Example 6 was changed to 2-(5-chloro-1H-indol-3-yl)acetic acid.

[0216] 1 H NMR (CDCl 3 ,500MHz): δ8.25(s,1H),7.82(t,J=8.6Hz,1H),7.74(t,J=8.7Hz,1H),7.46(s,1H),7.2 2(d,J=8.6Hz,1H),7.12(dd,J=8.6,1.6Hz,1H),7.06(s,1H),3.92(s,2H),3.43(s,3H).

[0217] 13 C NMR (CDCl 3 ,125MHz): δ159.06,157.93,156.01,151.70,151.60,142.09,134.54,128.35,125. 61,124.52,122.72,119.05,118.34,117.99,117.95,117.26,112.39,35.67,29.82.

[0218] 8. Synthesis of N-(Benzo[di]thiazol-2-yl)-2-(5-chloro-1-methyl-1H-indol-3-yl)-N-methylacetamide (Compound 51)

[0219] By the same experimental procedure as in Preparation Example 5, while also using N-(benzo[di]thiazol-2-yl)-2-(5-chloro-1H-indol-3-yl)acetamide, the white title compound (26.4 mg, 54%) was obtained.

[0220] 1 H NMR (CDCl 3 ,500MHz): δ7.83(d,J=8.1Hz,1H),7.80(d,J=7.9Hz,1H),7.58(d,J=1.0Hz,1H),7.43(t,J=7.6Hz,1H),7.30(t,J=7 .5Hz,1H),7.23(d,J=8.7Hz,1H),7.20(dd,J=8.7,1.5Hz,1H),7.08(s,1H),4.11(s,2H),3.87(s,3H),3.75(s,3H).

[0221] 13 C NMR (CDCl 3 ,125MHz): δ171.51,160.22,148.12,135.51,133.56,129.19,128.65,126.09,125. 58,124.03,122.55,121.42,121.23,118.28,110.71,105.57,35.73,33.16,32.49.

[0222] 9. Synthesis of N-(5-chloro-6-fluoropyridin-2-yl)-N-methyl-2-(1-methyl-1H-indol-3-yl)acetamide (Compound 36)

[0223] By the same experimental procedure as in Preparation Example 5, while also using N-(5-chloro-6-fluoropyridin-2-yl)-2-(1H-indol-3-yl)acetamide, the yellow title compound (7.2 mg, 54%) was obtained.

[0224] 1 H NMR (CDCl 3,500MHz): δ7.69(t,J=8.7Hz,1H),7.50(d,J=7.9Hz,1H),7.35(d,J=7.0Hz,1H),7.29(d,J=8.2Hz,1H ),7.23(t,J=7.6Hz,1H),7.11(t,J=7.4Hz,1H),6.95(s,1H),3.96(s,2H),3.75(s,3H),3.43(s,3H).

[0225] 13 C NMR (CDCl 3 ,125MHz): δ172.27,157.85,155.94,151.78,151.69,141.87,141.86,136.99,127.65,127.61, 122.00,119.36,118.82,118.03,117.98,112.96,112.72,109.46,107.06,35.56,32.87,32.82.

[0226] 10. Synthesis of 2-(5-chloro-1-methyl-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)-N-methylacetamide (Compound 37)

[0227] By the same experimental procedure as in Preparation Example 5, while also using 2-(5-chloro-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)acetamide, the yellow title compound (17.7 mg, 56%) was obtained.

[0228] 1 H NMR (CDCl 3 ,500MHz): δ7.74(t,J=8.7Hz,1H),7.44(s,1H),7.35(br s,1H),7.19(d,J=8.6Hz,1H),7.15(dd,J=8.7,1.4Hz,1H),6.98(s,1H),3.90(s,2H),3.72(s,3H),3.43(s,3H).

[0229] 13 C NMR (CDCl 3,125MHz): δ171.82,157.89,155.98,151.69,151.64,142.02,141.98,135.39,129.12,128.67, 125.28,122.27,118.36,117.98,117.94,113.23,112.96,110.54,106.88,35.61,33.06,32.43.

[0230] 11. Synthesis of 2-(5-chloro-1-methyl-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)acetamide (Compound 38)

[0231] By the same experimental procedure as in Preparation Example 5, while also using 2-(5-chloro-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)acetamide, a white title compound (8.20 mg, 27%) was obtained.

[0232] 1 H NMR (CDCl 3 ,500MHz): δ8.14(d,J=8.5Hz,1H),7.86(s,1H),7.76(t,J=8.8Hz,1H),7.51(s,1H),7.2 7(d,J=8.2Hz,1H),7.22(dd,J=8.7,1.5Hz,1H),7.09(s,1H),3.84(s,2H),3.81(s,3H).

[0233] 13 C NMR (CDCl 3 ,125MHz): δ169.98,157.71,155.80,147.54,147.44,142.93,135.85,129.96,128.47, 126.07,123.10,118.22,111.79,111.75,111.00,110.94,110.68,105.82,34.50,33.27

[0234] 12. Synthesis of N-(5-bromo-6-methylpyridin-2-yl)-2-(1-methyl-1H-indol-3-yl)acetamide (Compound 6)

[0235] [Process 5]

[0236]

[0237] While a solution of 2-(1-methyl-1H-indol-3-yl)acetic acid (200 mg, 1.06 mmol) in DMF (5 mL) was stirred at room temperature, 5-bromo-6-methylpyridin-2-amine (197 mg, 1.06 mmol), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide hexafluorophosphate (HATU, 402 mg, 1.06 mmol) and trimethylamine (0.3 mL, 2.11 mmol) were added in sequence. The reaction mixture was stirred at room temperature for 3 days. Distilled water was added to the mixture to terminate the reaction. The layers were separated with ethyl acetate, and the organic layer was washed with distilled water, washed with anhydrous Na 2 SO 4 Dried and filtered. After the filtrate was concentrated under reduced pressure, the concentrate was purified by column chromatography to give the title compound (120 mg, 31%).

[0238] 1 H NMR (CDCl 3 ,400MHz): δ7.99(m,2H),7.72(d,1H,J=12.0Hz),7.58(d,1H,J=12.0Hz),7.35(d,1H,J =8.0Hz),7.27(m,1H),7.15(m,1H),7.06(s,1H),3.87(s,2H),3.80(s,3H),2.42(s,3H)

[0239] 13. Synthesis of N-(5-bromo-6-methylpyridin-2-yl)-2-(1H-indol-3-yl)acetamide (Compound 5)

[0240] [Process 6]

[0241]

[0242] The title compound (110 mg, 30%) was obtained by the same experimental procedure as in Preparation Example 12, except that acetic acid in Preparation Example 12 was changed to 2-(1H-indol-3-yl)acetic acid.

[0243] 1 H NMR (CDCl 3 ,400MHz): δ8.37(s,1H),7.99(d,1H,J=12.0Hz),7.74(d,1H,J=8.0Hz),7.60(d,1H,J=8.0H z),7.40(d,1H,J=8.0Hz),7.25(m,1H),7.16(m,1H),3.87(s,2H),3.90(s,3H),2.43(s,3H)

[0244] 14. Synthesis of N-(Benzo[di]thiazol-2-yl)-2-(1H-indol-3-yl)acetamide (Compound 43)

[0245] [Process 7]

[0246]

[0247] While a solution of 2-(1H-indol-3-yl)acetic acid (100 mg, 0.57 mmol) in DMF (3 mL) was stirred at room temperature, benzo[di]thiazol-2-amine (197 mg, 0.57 mmol), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide hexafluorophosphate (HATU, 260 mg, 0.68 mmol) and trimethylamine (0.16 mL, 1.14 mmol) were added in sequence. The reaction mixture was stirred at room temperature for 3 days. Distilled water was added to the mixture to terminate the reaction. The layers were separated with ethyl acetate, and the organic layer was washed with distilled water, filtered through anhydrous Na 2 SO 4 Dried and filtered. After the filtrate was concentrated under reduced pressure, the concentrate was purified by column chromatography to give the title compound (10 mg, 6%).

[0248] 1 H NMR (CDCl 3 ,400MHz): δ8.99(s,1H),8.32(s,1H)7.80(d,1H,J=8.0Hz),7.64(d,1H,J=8.0H z),7.56(d,1H,J=12.0Hz),7.40(m,2H),7.29(m,3H),7.16(m,1H),4.03(s,2H)

[0249] 15. Synthesis of N-(5-chloro-6-fluoropyridin-2-yl)-2-(1H-indol-3-yl)acetamide (Compound 23)

[0250] [Process 8]

[0251]

[0252] The title compound (6 mg, 3%) was obtained by the same experimental procedure as in Preparation Example 14, except that the amine of Preparation Example 14 was changed to 5-chloro-6-fluoropyridin-2-amine.

[0253] 1 H NMR (CDCl 3,400MHz): δ8.14(dd,1H,J=8.0Hz and 2.0Hz),7.95(s,1H)7.75(m,1H),7.57(d,1H,J=8.0Hz),7.44(m,1H),7.24(m,2H),7.16(m,1H),3.92(s,2H)

[0254] 16. Synthesis of 2-(1H-indol-3-yl)-N-(3,4,5-trimethoxyphenyl)acetamide (Compound 71)

[0255] [Process 9]

[0256]

[0257] The title compound (10 mg, 5%) was obtained by the same experimental procedure as in Preparation Example 14, except that the amine in Preparation Example 14 was changed to 3,4,5-trimethoxyaniline.

[0258] 1 H NMR (DMSO-d6, 400MHz): δ7.59 (d, 1H, J = 8.0Hz), 7.35 (d, 1H, J = 8.0Hz), 7.25 (m, 1 H)7.07(m,1H),7.00(s,1H),6.97(m,1H),3.71(s,6H),3.69(s,2H),3.59(s,3H)

[0259] 17. Synthesis of 2-(5-chloro-1H-indol-3-yl)-N-(3,4,5-trimethoxyphenyl)acetamide (Compound 68)

[0260] [Process 10]

[0261]

[0262] While a solution of 2-(5-chloro-1H-indol-3-yl)acetic acid (100 mg, 0.47 mmol) in DMF (3 mL) was stirred at room temperature, 3,4,5-trimethoxyaniline (87 mg, 0.47 mmol), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide hexafluorophosphate (HATU, 217 mg, 0.57 mmol) and trimethylamine (0.13 mL, 0.95 mmol) were added in sequence. The reaction mixture was stirred at room temperature for 3 days. Distilled water was added to the mixture to terminate the reaction. The layers were separated with ethyl acetate, and the organic layer was washed with distilled water, washed with anhydrous Na 2 SO 4Dried and filtered. After the filtrate was concentrated under reduced pressure, the concentrate was purified by column chromatography to give the title compound (10 mg, 5%).

[0263] 1 H NMR (DMSO-d6, 400MHz): δ11.12 (s, 1H), 10.05 (s, 1H), 7.65 (d, 1H, J = 4.0Hz), 7.38 (s ,1H),7.36(s,1H),7.07(m,1H),6.99(s,2H),3.72(s,6H),3.68(s,2H),3.33(s,3H)

[0264] 18. Synthesis of N-(3,5-dichlorophenyl)-2-(1H-indol-3-yl)acetamide (Compound 81)

[0265] [Process 11]

[0266]

[0267] The title compound (8 mg, 5%) was obtained by the same experimental procedure as in Preparation Example 14, except that the amine in Preparation Example 14 was changed to 3,5-dichloroaniline.

[0268] 1 H NMR (CDCl 3 ,400MHz): δ8.50(s,1H),7.60(s,1H),7.55(d,1H,J=8.0Hz),7.41(d,1H,J=8.0 Hz),7.30(d,1H,J=4.0Hz),7.24(m,1H),7.16(m,2H),7.01(m,1H),3.86(s,2H)

[0269] 19. Synthesis of 2-(5-chloro-1H-indol-3-yl)-N-(3,5-dichlorophenyl)acetamide (Compound 78)

[0270] [Process 12]

[0271]

[0272] The title compound (10 mg, 6%) was obtained by the same experimental procedure as in Preparation Example 17, except that the amine in Preparation Example 17 was changed to 3,5-dichloroaniline.

[0273] 1 H NMR (CDCl 3,400MHz): δ8.33(s,1H),7.56(s,1H),7.36(d,1H,J=8.0Hz),7.34(d,1H,J =2.0Hz),7.30(bs,1H),7.26(m,1H),7.23(m,2H),7.06(m,1H),3.85(s,2H)

[0274] 20. Synthesis of 2-(5-chloro-1H-indol-3-yl)-N-(pyridin-4-yl)acetamide (Compound 88)

[0275] [Process 13]

[0276]

[0277] The title compound (10 mg, 7%) was obtained by the same experimental procedure as in Preparation Example 17, except that the amine of Preparation Example 17 was changed to pyridin-4-amine.

[0278] 1 H NMR(DMSO-d6,400MHz): δ11.15(s,1H),10.49(s,1H),8.40(m,1H),7.63(d,1H,J=2.0 Hz),7.57(m,1H)7.37(d,1H,J=8.0Hz),7.34(d,1H,J=4.0Hz),7.07(dd,1H,J=12.0Hz and 2.0Hz),3.72(s,6H),3.77(s,2H)

[0279] 21. Synthesis of N-(Benzo[di]thiazol-2-yl)-2-(1H-indol-3-yl)-N-methylacetamide (Compound 44)

[0280] [Process 14]

[0281]

[0282] While stirring a solution of 2-(1H-indol-3-yl)acetic acid (960 mg, 5.48 mmol) in DMF (35 mL) at room temperature, N-methylbenzo[di]thiazole-2-amine (600 mg, 3.65 mmol), N,N,N',N'-tetramethyl-O-(1H-benzotriazole-1-yl)uronium hexafluorophosphate (HBTU, 2.77 g, 7.31 mmol) and N,N-diisopropylethylamine (2.55 mL, 14.61 mmol) were added in sequence. The reaction mixture was stirred at room temperature for 3 days. Distilled water was added to the mixture to terminate the reaction. The layers were separated with ethyl acetate, and the organic layer was washed with distilled water, filtered through anhydrous Na 2 SO 4Dried and filtered. After the filtrate was concentrated under reduced pressure, the concentrate was purified by column chromatography to give the title compound (460 mg, 39%).

[0283] 1 H NMR(DMSO-d6,400MHz): δ11.02(s,1H),7.93(m,1H),7.79(m,1H)7.57(m,1H), 7.38(m,2H),7.31(m,2H),7.08(m,1H),6.98(m,1H),4.23(s,2H),3.84(s,3H)

[0284] 22. Synthesis of N-(Benzo[di]thiazol-2-yl)-2-(5-chloro-1H-indol-3-yl)-N-methylacetamide (Compound 47)

[0285] [Process 15]

[0286]

[0287] N-Methylbenzo[di]thiazol-2-amine (100 mg, 0.61 mmol) was stirred in CH 2 Cl 2 (12mL) solution, add triethylamine (0.65mL, 3.68mmol), and then further add 2-(5-chloro-1H-indol-3-yl)acetyl chloride (280mg, 1.23mmol). The reaction mixture was stirred at room temperature for 1 day. Distilled water was added to the mixture to terminate the reaction. The layers were separated, and the organic layer was washed with distilled water, washed with anhydrous Na 2 SO 4 Dried and filtered. After the filtrate was concentrated under reduced pressure, the concentrate was purified by column chromatography to give the title compound (30 mg, 7%).

[0288] 1 H NMR (DMSO-d6, 400MHz): δ11.22(s,1H),7.95(d,1H,J=8.0Hz),7.80(d,1H,J= 8.0Hz),7.66(d,1H,J=4.0Hz),7.42(m,3H)7.31(m,1H),7.09(dd,1H,J=8.0Hz and2.0Hz),4.25(s,2H),3.87(s,3H)

[0289] 23. Synthesis of N-(Benzo[di]thiazol-2-yl)-2-(1-methyl-1H-indol-3-yl)acetamide (Compound 54)

[0290] [Process 16]

[0291]

[0292] Benzo[di]thiazol-2-amine (214 mg, 1.43 mmol) was stirred in CH 2 Cl 2 (15mL) solution, add triethylamine (0.66mL, 4.75mmol), and then further add 2-(1-methyl-1H-indol-3-yl)acetyl chloride (329mg, 1.58mmol). The reaction mixture was stirred at room temperature for 1 day. Distilled water was added to the mixture to terminate the reaction. The layers were separated, and the organic layer was washed with distilled water, washed with anhydrous Na 2 SO 4 Dried and filtered. After the filtrate was concentrated under reduced pressure, the concentrate was purified by column chromatography to give the title compound (120 mg, 23%).

[0293] 1 H NMR (CDCl 3 ,400MHz): δ8.96(s,1H),7.80(d,1H,J=8.0Hz),7.63(d,1H,J=8.0Hz),7.54(d ,1H,J=8.0Hz),7.30(m,2H)7.16(m,1H),7.10(s,1H),4.01(s,2H),3.83(s,3H)

[0294] 24. Synthesis of N-(Benzo[di]thiazol-2-yl)-2-(5-fluoro-1H-indol-3-yl)-N-methylacetamide (Compound 60)

[0295] [Process 17]

[0296]

[0297] While stirring a solution of 2-(5-fluoro-1H-indol-3-yl)acetic acid (352 mg, 1.83 mmol) in DMF (12 mL) at room temperature, N-methylbenzo[di]thiazole-2-amine (200 mg, 1.22 mmol), N,N,N',N'-tetramethyl-O-(1H-benzotriazole-1-yl)uronium hexafluorophosphate (HBTU, 923 mg, 2.44 mmol) and N,N-diisopropylethylamine (0.9 mL, 4.87 mmol) were added in sequence. The reaction mixture was stirred at room temperature for 3 days. Distilled water was added to the mixture to terminate the reaction. The layers were separated with ethyl acetate, and the organic layer was washed with distilled water, filtered through anhydrous Na 2 SO 4Dried and filtered. After the filtrate was concentrated under reduced pressure, the concentrate was purified by column chromatography to give the title compound (140 mg, 33%).

[0298] 1 H NMR (DMSO-d6, 400MHz): δ11.12(s,1H),7.95(d,1H,J=8.0Hz),7.80(d,1H,J=8.0Hz),7.37(m,5H)6.93(m,1H),4.23(s,2H),3.86(s,3H)

[0299] 25. Synthesis of N-(Benzo[di]thiazol-2-yl)-2-(5-chloro-1-methyl-1H-indol-3-yl)acetamide (Compound 56)

[0300] [Process 18]

[0301]

[0302] While stirring a solution of 2-(5-chloro-1-methyl-1H-indol-3-yl)acetic acid (300 mg, 1.34 mmol) in DMF (13 mL) at room temperature, benzo[di]thiazole-2-amine (161 mg, 1.07 mmol), N,N,N',N'-tetramethyl-O-(1H-benzotriazole-1-yl)uronium hexafluorophosphate (HBTU, 1.02 g, 2.68 mmol) and N,N-diisopropylethylamine (0.9 mL, 5.37 mmol) were added in sequence. The reaction mixture was stirred at room temperature for 3 days. Distilled water was added to the mixture to terminate the reaction. The layers were separated with ethyl acetate, and the organic layer was washed with distilled water, filtered through anhydrous Na 2 SO 4 Dried and filtered. After the filtrate was concentrated under reduced pressure, the concentrate was purified by column chromatography to give the title compound (170 mg, 35%).

[0303] 1 H NMR (CDCl 3 ,400MHz): δ9.35(s,1H),7.81(d,1H,J=8.0Hz),7.65(d,1H,J=8.0Hz),7.4 8(m,1H),7.39(m,1H),7.25(m,3H),7.04(s,1H),3.94(s,2H),3.76(s,3H)

[0304] 26. Synthesis of N-(Benzo[di]thiazol-2-yl)-2-(5-fluoro-1H-indol-3-yl)acetamide (Compound 64)

[0305] [Process 19]

[0306]

[0307] While stirring a solution of 2-(5-fluoro-1H-indol-3-yl)acetic acid (50 mg, 0.25 mmol) in DMF (3 mL) at room temperature, benzo[di]thiazole-2-amine (31 mg, 0.20 mmol), N,N,N',N'-tetramethyl-O-(1H-benzotriazole-1-yl)uronium hexafluorophosphate (HBTU, 196 mg, 0.51 mmol) and N,N-diisopropylethylamine (0.2 mL, 1.04 mmol) were added in sequence. The reaction mixture was stirred at room temperature for 3 days. Distilled water was added to the mixture to terminate the reaction. The layers were separated with ethyl acetate, and the organic layer was washed with distilled water, filtered through anhydrous Na 2 SO 4 Dried and filtered. After the filtrate was concentrated under reduced pressure, the concentrate was purified by column chromatography to give the title compound (10 mg, 12%).

[0308] 1 H NMR (DMSO-d6, 400MHz): δ12.65 (s, 1H), 11.10 (s, 1H), 7.95 (d, 1H, J = 8.0Hz) ,7.74(d,1H,J=8.0Hz),7.38(m,4H),7.28(m,1H),6.93(m,1H),3.90(s,2H)

[0309] 27. Synthesis of N-(Benzo[di]thiazol-2-yl)-2-(6-chloro-1H-indol-3-yl)acetamide (Compound 41)

[0310] [Process 20]

[0311]

[0312] The title compound (6 mg, 3%) was obtained by the same experimental procedure as in Preparation Example 26, except that acetic acid in Preparation Example 26 was changed to 2-(6-chloro-1H-indol-3-yl)acetic acid.

[0313] 1 H NMR (MeOD-d4, 400MHz): δ11.65 (s, 1H), 11.10 (s, 1H), 7.85 (d, 1H, J = 8.0Hz), 7.74 (d, 1H,J=8.0Hz),7.58(d,1H,J=8.0Hz),7.43(m,2H),7.31(m,2H),7.05(dd,1H,J=8.0Hz and 4.0Hz),4.87(s,2H)

[0314] 28. Synthesis of 2-(5-chloro-1H-indol-3-yl)-N-(thiazol-2-yl)acetamide (Compound 99)

[0315] [Process 21]

[0316]

[0317] While stirring a solution of 2-(5-chloro-1H-indol-3-yl)acetic acid (125 mg, 0.49 mmol) in DMF (5 mL) at room temperature, thiazole-2-amine (50 mg, 0.59 mmol), N,N,N',N'-tetramethyl-O-(1H-benzotriazole-1-yl)uronium hexafluorophosphate (HBTU, 378 mg, 0.99 mmol) and N,N-diisopropylethylamine (0.4 mL, 2.00 mmol) were added in sequence. The reaction mixture was stirred at room temperature for 3 days. Distilled water was added to the mixture to terminate the reaction. The layers were separated with ethyl acetate, and the organic layer was washed with distilled water, filtered through anhydrous Na 2 SO 4 Dried and filtered. After the filtrate was concentrated under reduced pressure, the concentrate was purified by column chromatography to give the title compound (19 mg, 13%).

[0318] 1 H NMR (DMSO-d6, 400MHz): δ12.31 (s, 1H), 11.18 (s, 1H), 7.66 (d, 1H, J = 2.0Hz), 7.46 (d, 1H,J=4.0Hz),7.38(s,1H),7.36(m,1H),7.18(d,2H,J=4.0Hz),7.07(dd,1H,J=8.0Hz and 4.0Hz),3.84(s,2H)

[0319] 29. Synthesis of 2-(5-chloro-1H-indol-3-yl)-N-(quinolin-2-yl)acetamide (Compound 109)

[0320] [Process 22]

[0321]

[0322] The title compound (18 mg, 15%) was obtained by the same experimental procedure as in Preparation Example 28, except that the amine in Preparation Example 28 was changed to quinolin-2-amine.

[0323] 1H NMR(DMSO-d6,400MHz): δ11.16(s,1H),10.98(s,1H),8.30(m,1H),7.89(dd,1H,J=8.0Hz and 2.0Hz),7.82(d,1H,J=4.0Hz),7.72(m,2H),7.48(m,1H),7.40(d,1H,J=2.0Hz),7.37(d,1H,J=12.0Hz),7.07(dd,1H,J=8.0Hz and 4.0Hz),3.86(s,2H)

[0324] 30. Synthesis of 2-(5-chloro-1H-indol-3-yl)-N-(4,5,6,7-tetrahydrobenzo[di]thiazol-2-yl)acetamide (Compound 104)

[0325] [Process 23]

[0326]

[0327] The title compound (19 mg, 17%) was obtained by the same experimental procedure as in Preparation Example 28, except that the amine in Preparation Example 28 was changed to 4,5,6,7-tetrahydrobenzo[di]thiazol-2-amine.

[0328] 1 H NMR (DMSO-d6, 400MHz): δ12.07 (s, 1H), 11.17 (s, 1H), 7.63 (d, 1H, J = 4.0Hz), 7.37 (d, 1H, J = 8.0Hz and 2.0Hz), 7.07 (dd, 1H, J = 8.0Hz and 4.0Hz),3.78(s,2H),2.52(m,8H)

[0329] The structures of the above compounds are shown in Tables 2a to 2c, and the molecular weights of the compounds are listed in Table 2d.

[0330] [Table 2a]

[0331]

[0332] [Table 2b]

[0333]

[0334] [Table 2c]

[0335]

[0336] [Table 2d]

[0337] Compound No. Molecular weight Compound No. Molecular weight 6 358.23 88 285.73 5 344.21 44 321.40 8 378.65 47 355.84 43 307.37 54 321.40 40 341.81 60 339.39 23 303.72 56 355.84 26 338.16 64 325.36 71 340.37 41 341.81 68 374.82 99 291.76 81 319.19 109 335.79 78 353.63 104 345.85 2 362.20

[0338] Example: Measurement of the activity of compounds - Experimental protocol

[0339] 1. Search and preparation of compounds

[0340] In order to confirm the target specificity of the prepared compounds, evaluation was performed by the following method.

[0341] After HepG2 cells cultured in DMEM-fetal bovine serum (FBS) 10% medium were recovered and the survival rate was confirmed to be 97% or more by trypan blue staining, the recovered product was centrifuged at 1200 rpm for 5 minutes at room temperature, and the cells were centrifuged at 3×10 5 The cells were prepared by resuspending the cells at 100 cells / ml in DMEM-10% fetal bovine serum medium. Thereafter, the cells were dispensed into 60 mm culture dishes in 3 mL portions, and each dish was treated with 50 μl of a 5 μM compound diluted in DMEM medium, and then incubated in a cell culture incubator (5% CO 2 As a control, 50 μl of 0.05% dimethyl sulfoxide (DMSO) / DMEM medium was used for treatment.

[0342] Recover cultured cells to prepare mRNA samples. Specifically, mRNA was extracted from the recovered cells by phenol-chloroform precipitation using Trizol reagent (Invitrogen, catalog number 15596018). From the isolated RNA, cDNA was synthesized by reverse transcription, and the expression of CYP1A1 was confirmed by real-time polymerase chain reaction (PCR) using iQ SYBR-Green Supermix (Bio-rad) in the CFX96 (Bio-rad) detection system. GAPDH was used as a control enzyme to compare the relative values ​​of enzyme expression levels by the ΔΔct method. Herein, one (1) times was set using a control.

[0343] Real-time polymerase chain reaction was performed at an annealing temperature of 58° C. for 45 cycles using the following primer sequences.

[0344] Human CYP1A1 forward, 5'-CAC CCT CAT CAG TAA TGG TCA GA-3' (SEQ ID NO: 1) and reverse, 5'-AAC GTG CTT ATC AGG ACC TC-3' (SEQ ID NO: 2); human GAPDH forward, 5'-TGA TGA CAT CAAGAA GGT GG-3' (SEQ ID NO: 3) and reverse, 5'-TTA CTC CTT GGA GGC CAT GT-3' (SEQ ID NO: 4).

[0345] As a result, it can be seen that the expression level of CYP1A1 in the test compound is higher than that in the control (solvent), and it can be further seen that the expression of CYP1A1 is significantly induced ( Figure 1 and 2 ).

[0346] 2. Inhibitory effect on the production of inflammatory factor IL-6

[0347] In order to evaluate the inhibitory effect of the compounds according to the present invention on the production of macrophage IL-6, the following experiment was performed.

[0348] After THP-1 cells cultured in RPMI-fetal bovine serum (FBS) 10% + 2-ME (mercaptoethanol) medium were recovered and the survival rate was confirmed to be 97% or more by trypan blue staining, the recovered product was centrifuged at 1200 rpm for 5 minutes at room temperature, and the cells were centrifuged at 5×10 5 Cells were prepared by resuspending the cells at 100 cells / ml in RPMI-10% fetal bovine serum + 2-ME medium. Thereafter, the cells were dispensed into 24-well plates (3 wells per sample) in 500 μl portions, and then 0.5 μl of PMA was added to 200 ng / ml per well, and each well was treated with 10 μl of a 5 μM compound diluted in RPMI + 2ME medium. The cells were incubated in a cell culture incubator (5% CO 2 After incubation for 48 hours in a cell culture incubator (5% CO), 5 μl of LPS dissolved in dPBS was added at 100 ng / ml for treatment, and then the cells were incubated in a cell culture incubator (5% CO 2 incubator) for 24 hours.

[0349] As a control, 10 μl 0.05% dimethyl sulfoxide (DMSO) / RPMI culture medium was used for treatment. A new microtube was used to reclaim the culture medium of the cultured cells, and the cells were recovered with 1 mL Trizol (invitrogen) and stored at -80 ° C. The sample was diluted to 1 / 5 by placing 10 μl of the culture medium recovered and 40 μl of analysis dilution buffer in a FACS tube (BD falcon), and then the capture magnetic beads (capture bead) in each sample were vortexed, and then 1 μl of the vortexed solution and the capture magnetic bead diluent were taken. Then, 49 μl of capture magnetic bead diluent was added to prepare 50 μl of capture magnetic bead solution per sample. After the capture magnetic bead solution was mixed by vortexing, 50 μl of capture magnetic bead solution was placed in a FACS tube containing each sample, vortexed again, and left to stand at room temperature for 1 hour.

[0350] After 1 hour, 1 μl PE detection reagent and 49 μl PE detection reagent diluent were added to prepare 50 μl PE detection solution for each sample. After vortexing, 50 μl PE detection solution for each sample was added to the FACS tube containing the capture magnetic bead solution and the sample. After vortexing, the FACS tube was left to stand at room temperature for 1 hour. After 1 hour, 1 mL CBA washing buffer was added to each tube, centrifuged at 400 g for 5 minutes, and the supernatant was removed. After vortexing gently, 150 μl fixing buffer was added, vortexed gently, and then analyzed using flow cytometry.

[0351] Results, such as Figure 3 and Figure 4 As shown, the IL-6 produced by THP-1 stimulated by LPS was significantly reduced by treatment with the compound. Specifically, all the compounds of the present invention showed low results compared with the results of the control (vehicle), and this means that the compounds effectively inhibited the production of IL-6.

[0352] 3. Effect of regulatory T cell generation

[0353] In order to investigate the effect of the compounds according to the present invention on inducing immune tolerance, an in vitro regulatory T cell (Foxp3+Treg) generation experiment was performed as follows.

[0354] Human peripheral blood (AllCells) and phosphate buffered saline (PBS) were mixed at a ratio of 1:1 to prepare a mixture, and then the mixture was slowly placed so as not to mix into the upper layer of Histopaque (Sigma). After centrifugation at 350 g for 20 minutes, only the mononuclear cell layer in the middle layer was collected and washed with HBSS (Hanks' balanced salt solution, ) was washed. After washing again with MACS buffer (Miltenyi Biotec) to obtain T cells, positive selection was performed using CD4 microbeads (Miltenyi Biotec) (autoMACS separator, Miltenyi Biotec). T cells collected by the above method were separated by centrifugation at 5×10 5 10 μg / ml of anti-CD3 (eBioscience TM ) were dispensed into 48-well plates in 150 μl portions and incubated in a cell culture incubator (37°C, 5% CO 2 The cells were reacted in an incubator for 3 hours and washed with phosphate buffered saline to prepare a plate. 250 μl of the resuspended T cells were dispensed into the prepared plate and incubated with 2 μg / ml of anti-CD28 (eBioscience TM), 5ng / ml TGFβ-1 (R&Dsystems) and 50U / ml IL-2 (Miltenyi Biotec) were used to treat each well. 5μl of each compound at a concentration of 2.5μM diluted in RPMI+2ME medium was used for treatment, and then the cells were incubated in a cell culture incubator (37°C, 5% CO 2 The cells were cultured in an incubator for 7 days. As a control, 5 μl of 0.05% dimethyl sulfoxide (DMSO) / RPMI medium was used for treatment. After 7 days, in order to confirm the effect of generating regulatory T cells, the cultured cells were recovered to determine the presence of Foxp3 protein.

[0355] The recovered cells were placed in a 5 mL FACS tube (BD Falcon) and washed with 1 mL of phosphate buffered saline. The cells were resuspended in 0.1 mL of FACS buffer (0.1% NaN 3 , 1% FBS) and treated with 1 μg human immunoglobulin G (human IgG, Sigma) to prevent nonspecific binding of antibodies. After reacting at 4°C for 15 minutes, the cells were washed with FACS buffer. Then, 1 mL of fixation / permeabilization solution (eBioscience TM ) was added to the FACS tube containing each sample, and then reacted at 4°C for 1 hour. Thereafter, the product was washed with permeabilization buffer (eBioscience TM ) were washed twice. Then, 0.25 μg Foxp3 monoclonal antibody (eBioscience TM ) were treated, and then the samples were stained for 4 to 30 minutes. The cells were washed twice with permeabilization buffer, suspended in 0.3 mL of FACS buffer, and measured by flow cytometry.

[0356] As a result, it was confirmed that the generation of Foxp3+ regulatory T cells was promoted by treatment with test compound Nos. 5, 8, 43, 40, 23, 26, 71, 81, 2, 44, 47, 56, 64 and 41 (see Figure 5 ). It can be seen that the compound effectively induces the generation and proliferation of regulatory T cells.

[0357] 4. Confirmation of therapeutic effects on inflammatory bowel disease

[0358] In order to study the therapeutic effect of the compounds according to the present invention on inflammatory bowel disease, inflammatory bowel disease was induced in C57BL / 6 mice, and compounds (No. 8, 43, 40, 26, 44, 54 and 60) were administered as follows to evaluate the efficacy ( Figures 6 to 7 ).

[0359] On day 0 of the experiment, a 1.5% DSS solution prepared by dissolving DSS (dextran sulfate sodium, MP biomedicals, catalog number 160110) in 1.5% sterile distilled water was given to C57BL / 6 mice (8 weeks old, female, 18±2 g) for drinking for 7 days. The 1.5% DSS solution was replaced at intervals of 2 days. Sterile distilled water was provided for drinking from day 8 of the experiment. Body weight and severity index were measured at intervals of 2 days from day 0 of the experiment to confirm the onset of inflammatory bowel disease.

[0360] The compound at 20 mg / kg per mouse was completely dissolved in 10% (v / v) DMSO corresponding to the administered dose, then diluted in a Cremophor EL-phosphate buffered saline mixture to produce a final DMSO:Cremophor EL:phosphate buffered saline (1:1:8, v / v / v), and then 200 μl was orally administered daily for a total of 10 times from day 2 to day 11 of the experiment. The inflammatory bowel disease severity index was visually observed and recorded at 2-day intervals according to a severity index system divided into 0 to 10 levels.

[0361] According to the following items (Table 3), inflammatory bowel disease symptoms were evaluated by summing the scores of the three items.

[0362] [Table 3]

[0363]

[0364] As a result of the analysis, it was confirmed that the body weight of the solvent control group began to decrease from the 6th day of the experiment, decreased by more than 10% on the 10th day of the experiment, and 100% induced enteritis, accompanied by an increase in the severity index of more than 5. The mice in the solvent control group showed a severity index of 7.29±2.29 on the 10th day of the experiment when the severity index reached the maximum. On the other hand, on the 10th day of the experiment, the experimental group administered with 20 mg / kg of the compound No. 8, 43, 40, 26, 44, 54 or 60 of the present invention showed a statistically significant therapeutic effect compared with the solvent control group. In addition, comparison of the large intestine weight: length ratio (weight: length ratio, mg / cm) on the 15th day of the experiment demonstrated that in terms of morphology, intestinal inflammation could be significantly suppressed (compared with the solvent control group: *, p<0.05; **, p<0.01; ***, p<0.001, see Figure 6 and 7 ). Specifically, compounds No. 8, 43, 40, 26, 44, 54 and 60 of the present invention showed excellent anti-inflammatory effects when administered at an amount of 20 mg / kg.

[0365] On the 15th day of the experiment, the large intestine of the mice was removed to prepare mRNA samples. In order to extract mRNA, the large intestine tissue was ground with a homogenizer to obtain a homogenous suspension. From the homogenous suspension, easy-spin TM (DNA-free) Total RNA extraction kit (Intron biotechnology, catalog number 17221) was used to extract mRNA by phenol-chloroform precipitation. From the isolated RNA, cDNA was synthesized by reverse transcription, and then the expression of inflammatory cytokines was confirmed by real-time polymerase chain reaction (PCR) using iQ SYBR-Green Supermix (Bio-rad) in the CFX96 (Bio-rad) detection system. GAPDH was used as a control enzyme and the relative values ​​of enzyme expression levels were compared by the ΔΔct method. Herein, one (1) times was set using normal mouse colon as a control.

[0366] Real-time polymerase chain reaction was performed at an annealing temperature of 58° C., under conditions of 45 cycles, and using the following primer sequences.

[0367] mouse IL-1β forward, 5′-CTC GTG CTG TCG GAC CCA TAT-3′ (SEQ ID NO: 5) and reverse, 5′-TTG AAG ACA AAC CGC TTT TCC A-3′ (SEQ ID NO: 6);

[0368] mouse IL-6 forward, 5′-CAT GTT CTC TGC GAA ATC GTG G-3′ (SEQ ID NO: 7) and reverse, 5′-AAC GCA CTA GGT TTG CCG AGT A-3′ (SEQ ID NO: 8);

[0369] mouse IL-17A forward, 5′-TTT AAC TCC CTT GGC GCA AAA-3′ (SEQ ID NO: 9) and reverse, 5′-CTT TCC CTC CGC ATT GAC AC-3′ (SEQ ID NO: 10);

[0370] mouse TNF-α forward, 5′-CCA CAC CGT CAG CCG ATT TG-3′ (SEQ ID NO: 11) and reverse, 5′-CAC CCA TTC CCT TCA CAG AGC-3′ (SEQ ID NO: 12);

[0371] mouse IL-10 forward, 5′-CAA GGC AGT GGA GCA GGT GAA-3′ (SEQ ID NO: 13) and reverse, 5′-CGG AGA GAG GTA CAA ACG AGG TT-3′ (SEQ ID NO: 14);

[0372] mouse Foxp3 forward, 5′-CCC ATC CCC AGG AGT CTT G-3′ (SEQ ID NO: 15) and reverse, 5′-ACC ATG ACT AGG GGC ACT GTA-3′ (SEQ ID NO: 16);

[0373] Mouse GAPDH forward, 5'-TTC ACC ACC ATG GAG AAG GC-3' (SEQ ID NO: 17) and reverse, 5'-GGC ATG GAC TGT GGT CAT GA-3' (SEQ ID NO: 18).

[0374] Administration of compound No. 8, 40, 26 or 54 significantly reduced the expression levels of inflammatory cytokines IL-1β, IL-6, IL-17A and TNF-α in colorectal lesions compared with the solvent control group (compared with the solvent control group: **, p<0.01; ***, p<0.001, see Figure 8 In addition, the administration of compound No. 8, 40, 26 or 54 significantly increased the expression levels of immunomodulatory factors IL-10 and Foxp3 in colorectal lesions compared with the solvent control group (compared with the solvent control group: ***, p < 0.001, see Fig. 9 ). From these results, it can be seen that the compounds No. 8, 40, 26 and 54 of the present invention can significantly reduce the expression of inflammatory factors in the intestine and significantly increase the expression of immune regulatory factors in the intestine.

[0375] In order to investigate the mucosal healing effect of the compounds according to the present invention, inflammatory bowel disease was induced in C57BL / 6 mice, and the degree of restoration of intestinal epithelial barrier integrity was evaluated by administering the compounds (No. 40, 26, 54) as follows.

[0376] On day 0 of the experiment, a 1.5% DSS solution prepared by dissolving 1.5% DSS in sterile distilled water was given to C57BL / 6 mice (8 weeks old, female, 18±2 g) for drinking for 7 days. The 1.5% DSS solution was replaced at intervals of 2 days. Sterile distilled water was provided for drinking from day 8 of the experiment. From day 0 of the experiment, body weight and severity index were measured at intervals of 2 days to confirm the onset of inflammatory bowel disease.

[0377] In the compound No. 40, 26 or 54 administration group according to the present invention, 20 mg / kg of the compound per mouse was completely dissolved in 10% (v / v) DMSO corresponding to the administration dose, and then diluted in a Cremophor EL-phosphate buffered saline mixture to prepare a final DMSO:Cremophor EL:phosphate buffered saline (1:1:8, v / v / v), and then, from the 2nd day to the 9th day of the experiment, 200 μl of the prepared solution was orally administered daily for a total of 8 times.

[0378] The day before FITC-dextran administration, mice were deprived of water overnight. On the 10th day of the experiment, 600mg / kg FITC-dextran (fluorescein isothiocyanate-dextran, Sigma Aldrich, catalog number FD40) was diluted in phosphate buffered saline and orally administered to mice once with 200 μl. After oral administration 4 hours, fluorescence was measured in the serum extracted from the heart (fluorimeter, excitation 485-490nm, emission 528-530nm).

[0379] Administration of compound No. 40, 26 or 54 significantly reduced serum FITC-dextran compared with the solvent control group (compared with the solvent control group: ***, p<0.001, see Fig.10 ). From the results, it can be seen that the compounds No. 40, 26 and 54 of the present invention exhibit significant mucosal healing effects.

[0380] Therefore, compounds 8, 43, 40, 26, 44, 54 and 60 of the present invention have therapeutic effects when orally administered in a mouse model with inflammatory bowel disease. Therefore, these compounds can be suggested as a useful therapeutic strategy as a novel orally administered therapeutic agent for inflammatory bowel disease.

[0381] 5. Confirmation of the effect of preventing inflammation-induced colorectal cancer

[0382] In order to study the effect of the compounds according to the present invention on preventing inflammation-induced colorectal cancer, the medical efficacy was evaluated by administering Compound No. 40 and Compound No. 26 to the AOM / DSS-induced colorectal cancer mouse model in C57BL / 6 mice as follows: Fig.11 ). Azoxymethane (AOM), a carcinogen, did not cause colorectal cancer when given alone to mice. However, when DSS was used to induce inflammation, colorectal cancer occurred.

[0383] AOM (Sigma Aldrich, catalog number A5486) was diluted with physiological saline to a concentration of 10 mg / kg and then intraperitoneally administered 3 times at intervals of 7 days (experimental day 0, day 7, day 14). On the 7th day of the experiment, a 1.5% DSS solution prepared by dissolving 1.5% DSS in sterile distilled water was given to C57BL / 6 mice (8 weeks old, female, 18±2 g) for drinking for 7 days. In addition, the 1.5% DSS solution was replaced at intervals of 2 days. Sterile distilled water was provided for drinking from the 8th day of the experiment.

[0384] In the administration groups of compound No. 40 and 26 according to the present invention, 20 mg / kg of the compound per mouse was completely dissolved in 10% (v / v) of DMSO corresponding to the administration dose, respectively, and then diluted in a Cremophor EL-phosphate buffered saline mixture to prepare a final DMSO: Cremophor EL: phosphate buffered saline (1:1:8, v / v / v), and then 200 μl of the prepared solution was orally administered daily from the 7th day to the 21st day of the experiment, for a total of 14 times. When the body weight of the solvent control group was reduced by 15% compared with the 56th day of the experiment when the peak body weight was reached, the colorectal cancer preventive effect of the compound was confirmed.

[0385] On the 93rd day of the experiment, the large intestine of the mice was removed to confirm the occurrence of tumors in the large intestine. The number of tumors in the large intestine was 11.33±4.33 in the solvent control group, 3.00±2.00 in the case of compound No. 40, and 3.17±1.17 in the case of compound No. 26. Thus, the number of tumors was significantly reduced by administration of compound No. 40 and 26 compared with the solvent control group (compared with the solvent control group: **, p<0.01, see Fig.11 ).

[0386] Therefore, the compounds No. 40 and 26 of the present invention have an inhibitory effect on the development of colorectal cancer induced by inflammation. Therefore, these compounds can be suggested as a useful therapeutic strategy as a novel oral therapeutic agent for inflammatory bowel disease with a colorectal cancer preventive effect.

[0387] 6. Confirmation of the effectiveness of treatment for multiple sclerosis

[0388] In order to study the therapeutic effect of the compound according to the present invention on multiple sclerosis, autoimmune encephalomyelitis (EAE) was induced in C57BL / 6 mice and the therapeutic efficacy was evaluated by administering the compound (8, 43, 40, 26) Figures 12 to 14 ).

[0389] On day 0, myelin oligodendrocyte glycoprotein 35-55 (MOG 35-55, Peptron) (200 μg), heat-killed Mycobacterium tuberculosis (Difco, catalog number 231141) (500 μg) and adjuvant (complete Freund's adjuvant, Sigma Aldrich, catalog number F5506) were mixed together and then soaked for 7 minutes. After 100 μl of submerged peptide was subcutaneously injected into the bilateral abdomen of each C57BL / 6 mouse (7 weeks old, female, 17±2g), 100 μl of pertussis toxin (Sigma Aldrich, catalog number P2980) (200ng) was given to the tail intravenous injection.

[0390] On day 2 of the experiment, the same amount of pertussis toxin was administered intravenously. The mice were examined for immersion leaking from the injection site and visually observed from day 7 of the experiment to confirm the onset of multiple sclerosis.

[0391] In the groups treated with compound No. 8, 43, 40 and 26, 20 mg / kg of the compound per mouse was completely dissolved in 10% (v / v) DMSO corresponding to the administered dose, then diluted in a Cremophor EL-phosphate buffered saline mixture to prepare a final DMSO:Cremophor EL:phosphate buffered saline (1:1:8, v / v / v), and then 200 μl of the prepared solution was intraperitoneally administered daily from day 12 to day 17 of the experiment, for a total of 6 times. In the severity index system divided into 0-5 levels, the multiple sclerosis index was visually observed and recorded at intervals of 2 days from day 7 of the experiment, and the symptoms of autoimmune encephalomyelitis were index-evaluated according to the following items (see Table 4).

[0392] [Table 4]

[0393] score symptom 0 Asymptomatic 1 Tail weakness 2 Tail weakness and hind limb weakness 3 Hind limb paralysis 4 Paralysis of the hind limbs and weakness of the front limbs 5 Dead or dying

[0394] As a result of the analysis, it was confirmed that multiple sclerosis developed in all experimental groups from the 7th day of the experiment, and acute reactions with a severity index of 3.0 or more were induced in 100% of the groups on the 18th day of the experiment.

[0395] The severity index of mice in the solvent control group was 3.33±0.17 on day 18 of the experiment (which was the acute reaction period) and 3.33±0.17 on day 36 of the experiment (which was the chronic reaction period). In addition, the solvent control group showed a relapse-remitting pattern and a high severity index throughout the experiment.

[0396] On the other hand, the severity index of the compound treatment group was: 1.17±0.56 in the compound No.8 treatment group, 1.83±0.17 in the compound No.43 treatment group, 1.17±0.22 in the compound No.40 treatment group, and 1.33±0.56 in the compound No.26 treatment group on the 18th day of the experiment; and, 1.17±0.56 in the compound No.8 treatment group, 2.00±0.33 in the compound No.43 treatment group, 1.33±0.62 in the compound No.40 treatment group, and 1.33±0.22 in the compound No.26 treatment group on the 36th day of the experiment, showing a therapeutic effect of alleviating acute reactions and chronic reactions compared with the solvent control group.

[0397] In addition, it was confirmed that the groups treated with Compound No. 8, 43, 40 and 26 had a statistically significant therapeutic effect compared with the solvent control group from the 16th day of the experiment, and even after the administration was stopped, the statistically significant therapeutic effect was maintained compared with the solvent control group (compared with the solvent control group: ***, p < 0.001, see Fig.12 ). From the above results, it can be seen that administration of 20 mg / kg to mice showed excellent initial therapeutic effect and sustained relapse prevention effect.

[0398] On the 42nd day of the experiment, the spinal cord of mice was extracted to prepare mRNA samples. In order to extract mRNA, the spinal cord tissue was ground with a homogenizer to obtain a homogenous suspension. Trizol reagent (Invitrogen, catalog number 15596018) was used to extract mRNA from the homogenous suspension by phenol-chloroform precipitation. Then, cDNA was synthesized from the isolated RNA by reverse transcription, and the iQ SYBR-Green Supermix (Bio-rad) in the CFX96 (Bio-rad) detection system was used to study the expression of inflammatory cytokines by real-time polymerase chain reaction (PCR). GAPDH was used as a control enzyme to compare the relative values ​​of enzyme expression levels by the ΔΔct method. Herein, one (1) times was set using the WT mouse spinal cord as a control.

[0399] Real-time polymerase chain reaction was performed at an annealing temperature of 58° C., under conditions of 45 cycles, and using the following primer sequences.

[0400] mouse IFN-γ forward, 5′-ATG AAC GCT ACA CAC TGC ATC-3” (SEQ ID NO: 19) and reverse, 5′-CCA TCC TTT TGC CAG TTC CTC-3” (SEQ ID NO: 20);

[0401] mouse IL-17A forward, 5′-TTT AAC TCC CTT GGC GCA AAA-3” (SEQ ID NO: 21) and reverse, 5′-CTT TCC CTC CGC ATT GAC AC-3” (SEQ ID NO: 22);

[0402] mouse IL-1β forward, 5′-CTC GTG CTG TCG GAC CCA TAT-3” (SEQ ID NO: 23) and reverse, 5′-TTG AAG ACA AAC CGC TTT TCC A-3” (SEQ ID NO: 24);

[0403] mouse GAPDH forward, 5′-TTC ACC ACC ATG GAG AAG GC-3″ (SEQ ID NO: 25) and reverse, 5′-GGC ATG GAC TGT GGT CAT GA-3″ (SEQ ID NO: 26);

[0404] mouse IL-10 forward, 5′-CAA GGC AGT GGA GCA GGT GAA-3′ (SEQ ID NO: 27) and reverse, 5′-CGG AGA GAG GTA CAA ACG AGG TT-3″ (SEQ ID NO: 28);

[0405] Mouse Foxp3 forward, 5'-CCC ATC CCC AGG AGT CTT G-3" (SEQ ID NO: 29) and reverse, 5'-ACC ATG ACT AGG GGC ACT GTA-3" (SEQ ID NO: 30).

[0406] In addition, the expression levels of inflammatory cytokines IFN-γ, IL-17A and IL-1β in spinal cord lesions were significantly reduced by administration of compounds No. 8, 43, 40 and 26 compared with the solvent control group (compared with the solvent control group: *, p<0.05; **, p<0.01; ***, p<0.001, see Fig.13 ). By administering compounds No.8, 43, 40 and 26, the expression levels of immunomodulatory factors IL-10 and Foxp3 in spinal cord lesions were significantly increased compared with the solvent control group (compared with the solvent control group: *, p<0.05; **, p<0.01, see Fig.14 ).

[0407] Therefore, the compounds No. 8, 43, 40 and 26 of the present invention have therapeutic efficacy in the multiple sclerosis mouse model, and the relapse prevention effect is continuously maintained even after the administration is stopped. Therefore, these compounds can be suggested as a useful therapeutic strategy for multiple sclerosis as a novel oral therapeutic agent.

[0408] 7. Confirmation of the therapeutic effect of graft-versus-host disease

[0409] In order to study the inhibitory effect of the compounds of the present invention on graft-versus-host disease (GVHD), acute graft-versus-host disease was induced in C57BL / 6 mice by allogeneic bone marrow transplantation as follows, and the compound (No. 40 or 26) was administered to evaluate its efficacy ( Fig.15 and 16 ).

[0410] The spleen of Balb / c IFN-γ knockout mice (8 to 12 weeks old, female, 18 ± 3 g) was removed, pulverized by adding RPMI medium and then passed through a 40 μm cell strainer (BD Falcon) to obtain a single cell suspension. The single cell suspension was centrifuged (1200 rpm, 5 minutes), and after discarding the supernatant, 1 mL of ACK (ammonium chloride / potassium bicarbonate) lysis buffer (0.15 M NH 4 Cl, 1 mM KHCO 3 , 0.1 mM Na 2 EDTA), followed by stirring for 1 min, and then washing with RPMI medium.

[0411] After centrifugation, the cell suspension was reacted on mouse CD90.2 microbeads (Miltenyi Biotec, catalog number 130-121-278) at 4°C for 20 minutes. After the reaction was completed, the cell suspension was centrifuged and washed with 10 mL Running buffer (Miltenyi Biotec, catalog number 130-091-221) was then used to wash with 3 mL Then, CD90.2 was obtained from the cell suspension using Auto MACS pro (Miltenyi Biotec). + T cells (positive selection). + The bone marrow cells transplanted together with T cells were aseptically obtained from the femur and tibia of wild-type Balb / c mice (8-12 weeks old, female, 18 ± 3g). The ends of the femur and tibia were cut off, and the bone marrow was extracted by perfusing RPMI culture medium to bone tissue with a syringe (femur 21G, tibia 26G). The extracted bone marrow was passed through a 40 μm cell strainer to obtain a single cell suspension.

[0412] The bone marrow single cell suspension was centrifuged, and after discarding the supernatant, 500 μl of ACK lysis buffer was added, followed by stirring for 30 seconds and washing the solution with RPMI medium. After centrifugation, the suspension was reacted on mouse CD90.2 microbeads at 4°C for 20 minutes. After the reaction was completed, the cell suspension was centrifuged and washed with 10 mL of Wash with running buffer and then 3 mL Then, CD90.2 was obtained from the cell suspension by Auto MACS pro. - T cell-depleted bone marrow cells (TCD-BM) (negative selection) were washed with phosphate-buffered saline to obtain IFN-γ knockout CD90.2 + T cells and normal TCD-BM. T cells were isolated at a concentration of 1 × 10 7 T cells were prepared by resuspending TCD-BM at 5 × 10 7 The cells were resuspended in phosphate-buffered saline to prepare TCD-BM.

[0413] Normal C57BL / 6 mice (9 to 11 weeks old, female, 19±3 g) were irradiated with 850 cGy of radiation at 3-hour intervals using a radiation irradiator. + The graft prepared by mixing T cells and TCD-BM in a ratio of 1:1 was injected into the tail vein of C57BL / 6 mice in an amount of 100 μl. In the administration group of compound No.40 or 26 according to the present invention, 20 mg / kg of the compound per mouse was completely dissolved in 10% (v / v) DMSO corresponding to the administration dose, and then diluted in a Cremophor EL-phosphate buffered saline mixture to prepare a final DMSO: Cremophor EL: phosphate buffered saline (1:1:8, v / v / v), and then 200 μl of the solution was intraperitoneally administered daily from the 4th day to the 9th day after transplantation, for a total of 6 times. The severity index of graft-versus-host disease was evaluated by visual observation at intervals of 2 days in the following severity index system, which is divided into a total of 10 points, wherein each item is 0 to 2 points, including weight loss, hair condition, posture, activity and skin changes.

[0414] As a result of the analysis, on the 7th day after transplantation, graft-versus-host disease was induced in 100% of the mice in the solvent control group, and the severity index was 4.12±1.20. In addition, on the 14th day after transplantation, the severity index was: 6.20±1.20 in the solvent control group; 0.60±0.60 in the compound No.40 treatment group; and 2.80±0.80 in the compound No.26 treatment group, showing significantly alleviated graft-versus-host disease and therapeutic effect compared with the solvent control group (compared with the solvent control group: ***, p<0.001, see Fig.15 ).

[0415] On the 15th day of the experiment, the lungs of mice were removed to prepare mRNA samples. In order to extract mRNA, spinal cord tissue was ground with a homogenizer to obtain a homogenous suspension. Trizol reagent (Invitrogen, catalog number 15596018) was used to extract mRNA from the homogenous suspension by phenol-chloroform precipitation. Then, cDNA was synthesized from the isolated RNA by reverse transcription, and the expression of inflammatory cytokines was confirmed by real-time polymerase chain reaction (PCR) using iQ SYBR-Green Supermix (Bio-rad) in the CFX96 (Bio-rad) detection system. GAPDH was used as a control enzyme to compare the relative values ​​of enzyme expression levels by the ΔΔct method. Herein, one (1) times was set using normal mouse spinal cord as a control.

[0416] Real-time polymerase chain reaction was performed at an annealing temperature of 58° C., under conditions of 45 cycles, and using the following primer sequences.

[0417] mouse IL-17A forward, 5′-TTT AAC TCC CTT GGC GCA AAA-3′ (SEQ ID NO: 31) and reverse, 5′-CTT TCC CTC CGC ATT GAC AC-3′ (SEQ ID NO: 32);

[0418] mouse IL-6 forward, 5′-CAT GTT CTC TGC GAA ATC GTG G-3′ (SEQ ID NO: 33) and reverse, 5′-AAC GCA CTA GGT TTG CCG AGT A-3′ (SEQ ID NO: 34);

[0419] mouse IL-10 forward, 5′-CAA GGC AGT GGA GCA GGT GAA-3′ (SEQ ID NO: 35) and reverse, 5′-CGG AGA GAG GTA CAA ACG AGG TT-3′ (SEQ ID NO: 36);

[0420] Mouse GAPDH forward, 5'-TTC ACC ACC ATG GAG AAG GC-3' (SEQ ID NO:37) and reverse, 5'-GGC ATG GAC TGT GGT CAT GA-3' (SEQ ID NO:38).

[0421] By administering compounds No. 40 and 26, the expression levels of inflammatory cytokines IL-6 and IL-17A in lung tissue were significantly reduced compared with the solvent control group. In addition, by administering compounds No. 40 and 26, the expression level of immunomodulatory factor IL-10 in lung tissue was significantly increased compared with the solvent control group (compared with the solvent control group: **, p < 0.01; ***, p < 0.001, see Fig.16 ).

[0422] Therefore, the compounds No. 40 and 26 of the present invention have therapeutic efficacy in a mouse model with graft-versus-host disease, and even after the administration is stopped, the therapeutic efficacy is continuously maintained due to the increase of immunomodulatory factors. Therefore, these compounds can be suggested as a useful therapeutic strategy for novel oral therapeutic agents for graft-versus-host disease. <110> Parenchyma Biotech Inc. <120> Compounds and pharmaceutical compositions for treating autoimmune diseases <130> 20P09015 <150> US 62 / 910,537 <151> 2019-10-04 <160> 38 <170> KoPatentIn 3.0 <210> 1 <211> twenty three <212> RNA <213> Homo sapiens <400> 1 caccctcatc agtaatggtc aga 23 <210> 2 <211> 20 <212> RNA <213> Homo sapiens <400> 2 aacgtgctta tcaggacctc 20 <210> 3 <211> 20 <212> RNA <213> Homo sapiens <400> 3 tgatgacatc aagaaggtgg 20 <210> 4 <211> 20 <212> RNA <213> Homo sapiens <400> 4 ttactccttg gaggccatgt 20 <210> 5 <211> twenty one <212> RNA <213> House mouse (Mus musculus) <400> 5 ctcgtgctgt cggacccata t 21 <210> 6 <211> twenty two <212> RNA <213> House mouse <400> 6 ttgaagacaa accgcttttc ca 22 <210> 7 <211> twenty two <212> RNA <213> House mouse <400> 7 catgttctct gcgaaatcgt gg 22 <210> 8 <211> twenty two <212> RNA <213> House mouse <400> 8 aacgcactag gtttgccgag ta 22 <210> 9 <211> twenty one <212> RNA <213> House mouse <400> 9 tttaactccc ttggcgcaaa a 21 <210> 10 <211> 20 <212> RNA <213> House mouse <400> 10 ctttccctcc gcattgacac 20 <210> 11 <211> 20 <212> RNA <213> House mouse <400> 11 ccacaccgtc agccgatttg 20 <210> 12 <211> twenty one <212> RNA <213> House mouse <400> 12 cacccattcc cttcacagag c 21 <210> 13 <211> twenty one <212> RNA <213> House mouse <400> 13 caaggcagtg gagcaggtga a 21 <210> 14 <211> twenty three <212> RNA <213> House mouse <400> 14 cggagagagg tacaaacgag gtt 23 <210> 15 <211> 19 <212> RNA <213> House mouse <400> 15 cccatcccca ggagtcttg 19 <210> 16 <211> twenty one <212> RNA <213> House mouse <400> 16 accatgacta ggggcactgt a 21 <210> 17 <211> 20 <212> RNA <213> House mouse <400> 17 ttcaccacca tggagaaggc 20 <210> 18 <211> 20 <212> RNA <213> House mouse <400> 18 ggcatggact gtggtcatga 20 <210> 19 <211> twenty one <212> RNA <213> House mouse <400> 19 atgaacgcta cacactgcat c 21 <210> 20 <211> twenty one <212> RNA <213> House mouse <400> 20 ccatcctttt gccagttcct c 21 <210> twenty one <211> twenty one <212> RNA <213> House mouse <400> twenty one tttaactccc ttggcgcaaa a 21 <210> twenty two <211> 20 <212> RNA <213> House mouse <400> twenty two ctttccctcc gcattgacac 20 <210> twenty three <211> twenty one <212> RNA <213> House mouse <400> twenty three ctcgtgctgt cggacccata t 21 <210> twenty four <211> twenty two <212> RNA <213> House mouse <400> twenty four ttgaagacaa accgcttttc ca 22 <210> 25 <211> 20 <212> RNA <213> House mouse <400> 25 ttcaccacca tggagaaggc 20 <210> 26 <211> 20 <212> RNA <213> House mouse <400> 26 ggcatggact gtggtcatga 20 <210> 27 <211> twenty one <212> RNA <213> House mouse <400> 27 caaggcagtg gagcaggtga a 21 <210> 28 <211> twenty three <212> RNA <213> House mouse <400> 28 cggagagagg tacaaacgag gtt 23 <210> 29 <211> 19 <212> RNA <213> House mouse <400> 29 cccatcccca ggagtcttg 19 <210> 30 <211> twenty one <212> RNA <213> House mouse <400> 30 accatgacta ggggcactgt a 21 <210> 31 <211> twenty one <212> RNA <213> House mouse <400> 31 tttaactccc ttggcgcaaa a 21 <210> 32 <211> 20 <212> RNA <213> House mouse <400> 32 ctttccctcc gcattgacac 20 <210> 33 <211> twenty two <212> RNA <213> House mouse <400> 33 catgttctct gcgaaatcgt gg 22 <210> 34 <211> twenty two <212> RNA <213> House mouse <400> 34 aacgcactag gtttgccgag ta 22 <210> 35 <211> twenty one <212> RNA <213> House mouse <400> 35 caaggcagtg gagcaggtga a 21 <210> 36 <211> twenty three <212> RNA <213> House mouse <400> 36 cggagagagg tacaaacgag gtt 23 <210> 37 <211> 20 <212> RNA <213> House mouse <400> 37 ttcaccacca tggagaaggc 20 <210> 38 <211> 20 <212> RNA <213> House mouse <400> 38 ggcatggact gtggtcatga 20

Claims

1. A compound represented by the following formula 1 or a pharmaceutically acceptable salt thereof: [Formula 1] Wherein A and R1 to R6 are listed in Table T1 below: [Table T1] 2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of the following compounds: N-(5-bromo-6-methylpyridin-2-yl)-2-(5-chloro-1H-indol-3-yl)acetamide; N-(Benzo[d]thiazol-2-yl)-2-(5-chloro-1H-indol-3-yl)acetamide; and 2-(5-Chloro-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)acetamide.

3. Use of a compound represented by the following formula 1 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating an autoimmune disease, wherein the autoimmune disease is any one selected from the group consisting of multiple sclerosis, inflammatory bowel disease, graft-versus-host disease, asthma, atopy, psoriasis, rheumatoid arthritis, systemic lupus erythematosus and type 1 diabetes: [Formula 1] Wherein A and R1 to R6 are listed in Table T2 below: [Table T2] 4. The use according to claim 3, wherein the compound is selected from the group consisting of: N-(5-bromo-6-methylpyridin-2-yl)-2-(5-chloro-1H-indol-3-yl)acetamide; N-(Benzo[d]thiazol-2-yl)-2-(5-chloro-1H-indol-3-yl)acetamide; 2-(5-chloro-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)acetamide; and N-(Benzo[d]thiazol-2-yl)-2-(1H-indol-3-yl)acetamide.

5. Use of a compound represented by the following formula 1 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating colon cancer: [Formula 1] Wherein A and R1 to R6 are listed in Table T2 below: [Table T2] 6. The use according to claim 5, wherein the compound is selected from the group consisting of: N-(5-bromo-6-methylpyridin-2-yl)-2-(5-chloro-1H-indol-3-yl)acetamide; N-(Benzo[d]thiazol-2-yl)-2-(1H-indol-3-yl)acetamide; N-(Benzo[d]thiazol-2-yl)-2-(5-chloro-1H-indol-3-yl)acetamide; and 2-(5-Chloro-1H-indol-3-yl)-N-(5-chloro-6-fluoropyridin-2-yl)acetamide.

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