Aryl receptor modulators and methods of making and using the same

Abstract

The present invention relates generally to compounds capable of binding to and modulating the activity of the aryl hydrocarbon receptor, methods of using these compounds to treat inflammation such as Crohn's disease, and pharmaceutical compositions comprising these compounds. Also provided are methods of increasing IL-22 levels and / or decreasing IFN-g levels in a subject.

Description

[0001] This application is a divisional application of Chinese patent application No. 202010710069.9, filed on June 26, 2015, entitled "Aromatic Receptor Modulator and Preparation and Use Method Thereof".

[0002] Related applications

[0003] This application claims the benefits of U.S. Provisional Application No. 62 / 017,959 (filed June 27, 2014) and U.S. Provisional Application No. 62 / 056,054 (filed September 26, 2014), each of which is incorporated herein by reference in its entirety. Background of the Invention

[0004] Aryl hydrocarbon receptors (AhRs) are basic helical-loop-helical proteins and members of the Per-ARNT-Sim (PAS) superfamily of proteins. Physiologically, many of these proteins function by sensing molecular and stimuli from the cellular / tissue microenvironment, thereby initiating necessary signaling cascades to elicit appropriate cellular responses.

[0005] In its inactivated state, AhR resides in the cytosol, binding to several co-protein chaperones. Upon activation, however, it migrates to the nucleus and binds to its dimer partner, ARNT (another bHLH-PAS protein), thereby initiating transcription of multiple genes with promoters containing dioxin (DRE) or heterogeneous conserved sequences (XME). Pioneer studies in AhR-deficient mice have highlighted the role of AhR in the development and function of multiple organs. Recent research has shown that AhR controls specific immune responses (see, e.g., Stockinger et al., Semin. Immunol. 2011, 23, 99-105).

[0006] AhR is highly expressed by T cells and controls Th1 / Th2 / Th17 cell-associated immunity. In humans and mouse models of inflammatory bowel disease, AhR activation leads to attenuation of IFN-γ and T-bet expression, key transcription factors driving Th1 polarization. The fundamental mechanism by which AhR inhibits IFN-γ production remains to be determined; however, AhR signaling has been shown to enhance Aiolos, a member of the Ikaros family, which negatively regulates IFN-γ production and colitis in mice. AhR activation in lymphocytes can also regulate the production of interleukin-22 (IL-22), a cytokine that exerts protective effects in multiple organs, and time-course studies have shown that inhibition of IFN-γ and T-bet occurs prior to IL-22 induction.

[0007] Given that selective AhR modulators may potentially affect immunity and inflammation, thereby treating various inflammatory conditions, there is a need for effective selective compounds that modulate AhR activity. Invention Overview

[0008] This disclosure, in one embodiment, provides a compound that effectively and selectively binds to AhR. In one aspect, this disclosure provides a compound according to formula I:

[0009]

[0010] The variables are defined below.

[0011] In one aspect, this disclosure relates to compounds according to formula IA:

[0012]

[0013] The variables are defined below.

[0014] In one aspect, this disclosure relates to compounds according to formula II:

[0015]

[0016] The variables are defined below.

[0017] In one aspect, this disclosure relates to compounds according to formula III:

[0018]

[0019] The variables are defined below.

[0020] In one aspect, this disclosure relates to compounds according to formula IV:

[0021]

[0022] The variables are defined below.

[0023] This disclosure also provides methods for treating inflammatory diseases or conditions, including administering a therapeutically effective amount of the above-mentioned compound to a subject in need. In some embodiments, the inflammatory disease or condition is selected from inflammatory bowel disease, chondritis, osteomalacia, ulcerative colitis, psoriasis, arthritis, psoriatic arthritis, rheumatoid arthritis, juvenile arthritis, juvenile rheumatoid arthritis, autoimmune hepatitis, Crohn's disease, lupus erythematosus, multiple sclerosis, Alzheimer's disease, dermatitis, atopic dermatitis, acne, type I diabetes, Raynaud's phenomenon, Graves' disease, and Addison's disease.

[0024] In some embodiments, the inflammatory disease or condition is selected from Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, diverticulitis, Becton's disease (BSC). Diseases including idiopathic inflammatory bowel disease, irritable bowel syndrome, regional enteritis, spastic colon, microscopic colitis, Crohn's colitis, perianal diseases, indeterminate colitis, lymphocytic gastritis, and eosinophilic enteritis.

[0025] In some embodiments, the inflammatory disease or condition is Crohn's disease. In some embodiments, Crohn's disease is selected from ileocolitis, ileitis, gastroduodenal Crohn's disease, jejunoileitis, and granulomatous ileocolitis. In some embodiments, Crohn's disease includes intestinal fibrosis. In some embodiments, Crohn's disease is fibrotic stricture Crohn's disease.

[0026] In another aspect, this disclosure provides methods for preventing, treating, or reducing fibrotic stenosis or intestinal fibrosis in subjects, comprising administering a therapeutically effective amount of the disclosed compound to a subject in need. In some embodiments, the fibrotic stenosis or intestinal fibrosis is associated with Crohn's disease.

[0027] In another aspect, the present invention provides compounds as described above for use as pharmaceuticals. The present invention further provides a method for using the above-described compounds to treat inflammatory diseases or conditions. The method comprises administering a therapeutically effective amount of the compound to a subject in need. In some embodiments, the inflammatory disease or condition is selected from inflammatory bowel disease, chondritis, osteomalacia, ulcerative colitis, psoriasis, arthritis, psoriatic arthritis, rheumatoid arthritis, juvenile arthritis, juvenile rheumatoid arthritis, autoimmune hepatitis, Crohn's disease, lupus erythematosus, multiple sclerosis, Alzheimer's disease, dermatitis, atopic dermatitis, acne, type I diabetes, Raynaud's phenomenon, Graves' disease, and Addison's disease. In some embodiments, the inflammatory disease or condition is selected from Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, diverted colitis, Behcet's disease, idiopathic inflammatory bowel disease, irritable bowel syndrome, regional enteritis, spastic colon, microscopic colitis, Crohn's colitis, perianal disease, indeterminate colitis, lymphocytic gastritis, and eosinophilic enteritis. In some embodiments, the inflammatory disease or condition is Crohn's disease. In some embodiments, Crohn's disease is selected from ileocolitis, ileitis, gastroduodenal Crohn's disease, jejunoileitis, and granulomatous ileocolitis. In some embodiments, Crohn's disease includes intestinal fibrosis. In some embodiments, Crohn's disease is fibrotic stricture Crohn's disease.

[0028] In another aspect, the present invention provides the use of the above-described compound in a method for preventing, treating, or reducing fibrostenosis or intestinal fibrosis in a subject. The method comprises administering an effective amount of the compound to a subject in need. In some embodiments, the fibrostenosis or intestinal fibrosis is associated with Crohn's disease. Brief description of the attached diagram

[0029] Figure 1 It shows a series of 1-aryl-1,2,3,4-tetrahydro-β-carboline derivatives.

[0030] Figure 2 Modifications to the 1-aryl-1,2,3,4-tetrahydro-β-carboline skeleton were shown to investigate SAR and provide improved properties.

[0031] Figure 3 Modifications to the 1-aryl-1,2,3,4-tetrahydro-β-carboline skeleton were shown to investigate SAR and provide improved properties.

[0032] Figure 4 Modifications to the 1-aryl-1,2,3,4-tetrahydro-β-carboline skeleton were shown to investigate SAR and provide improved properties.

[0033] Figure 5 Modifications to the leflunomide framework were shown to study SAR and provide improved properties.

[0034] Figure 6 The chemical structures of various compounds used for evaluating ADMET and other properties in this disclosure are shown.

[0035] Figure 7 The chemical structures of various compounds used for evaluating ADMET and other properties in this disclosure are shown.

[0036] Figure 8 The chemical structures of various compounds used for evaluating ADMET and other properties in this disclosure are shown.

[0037] Figure 9 The chemical structures of various compounds used for evaluating ADMET and other properties in this disclosure are shown.

[0038] Figure 10The calculated bioavailability of the compounds disclosed herein is displayed. All ADME data were calculated using ADME software suite v4.95.3. Human oral bioavailability (%F) is the portion of the compound that reaches systemic circulation after oral administration. A drug must meet the following requirements to be bioavailable: dissolve in the stomach or intestine under varying pH conditions; withstand acid hydrolysis at pH <2; permeate the intestinal membrane via passive or active transport; withstand efflux of P-gp in synergy with metabolic enzymes in the intestine; and withstand first-pass metabolism in the liver.

[0039] Figure 11 The calculated water solubility (logarithmic Sw) of various compounds disclosed herein is displayed. The solubility (logarithmic Sw) of compounds in pure water at 25°C and at various important physiological pH values ​​is predicted.

[0040] Figure 12 The pH-dependent solubility of the various compounds disclosed herein is shown.

[0041] Figure 13 The passive absorption in the human gut for the various compounds of this disclosure is shown. Human intestinal permeability is assessed by taking into account permeability across cellular and paracellular pathways, estimating the maximum intestinal passive absorption of the compounds. Absorption-related properties, such as permeability of the jejunum and Caco-2, and values ​​for absorption rate (ka) are also calculated using constants for lipophilicity (logarithm of P) and ionization (pKa value).

[0042] Figure 14 The calculations show active transport across the intestinal barrier for various compounds of this disclosure. The calculated transports include estimates of substrate predictions for PepT1 (oligopeptide transporter) and ASBT (bile acid transporter).

[0043] Figure 15 The algorithm displays the calculated P-glycoprotein specificity for various compounds disclosed herein. It identifies P-gp substrates and / or inhibitors. Substrates are compounds that are transported (exported) by P-gp. Inhibitors are compounds that block the transport of standard substrates (calcein and others) by P-gp.

[0044] Figure 16 The software displays the calculated tissue distribution of various compounds of this disclosure. It predicts the degree of protein binding in plasma to obtain the percentage of compounds circulating in their free pharmaceutically active form and calculates the apparent volume of distribution to estimate the distribution of compounds between plasma and body tissues.

[0045] Figure 17The software displays calculated interactions with cytochrome P450 for various compounds of this disclosure. The software calculates how the compounds interact with five cytochrome P450 (CYP) isomers: 3A4, 2D6, 2C9, 2C19, and 1A2, which are responsible for most metabolic reactions.

[0046] Figure 18 The data displayed, calculated using Tox software v2.95, represent the probability of genotoxicity from various compounds disclosed herein. Genotoxicity = probability of a positive Ames test. The Ames test is the most popular test for assessing the mutagenic properties of compounds. It is a short-term bacterial reverse mutagenesis test. This test is performed on various strains of Salmonella typhimurium and Escherichia coli. The Ames test is widely used worldwide as a primary screening tool to determine the mutagenic properties of NCEs in the pharmaceutical and chemical industries.

[0047] Figure 19 The data presented indicate the calculated probability of various compounds in this disclosure inhibiting hERG. Research on hERG potassium channel inhibition is forming an emerging field in drug safety studies. Drug interactions with hERG channels can lead to chronic QT syndrome, characterized by torsades de pointes, which can result in occasional death due to ventricular fibrillation. Several promising drugs have been withdrawn from the market in recent years due to several cases of sudden cardiac death caused by hERG channel inhibition. Therefore, early identification of potential safety concerns is extremely important to prevent the failure of costly research and development projects.

[0048] Figure 20 This displays the calculated probabilities of health effects for the various compounds disclosed herein. The predictions of health effects are based on long-term toxicity studies combined with reported adverse effects on specific organs or organ systems.

[0049] Figure 21 The calculated LD values ​​for the various compounds of this disclosure are shown. 50 Value. LD 50 The value can be regarded as an indicator of the "cumulative potential" to cause a variety of acute effects and death in animals, and is the most widely used measure of the "acute systemic toxicity" of a chemical.

[0050] Figure 22 This demonstrates the effects of various compounds of this disclosure at a final concentration of 200 nM on IL-22 and IFN-γ levels.

[0051] Figure 23 The effects of various compounds disclosed herein on IL-22 and IFN-γ levels, as well as the chemical structure of the known AhR conjugate Ficz, are shown.

[0052] Figure 24 This illustrates the effects of various compounds of this disclosure on IL-22 and IFN-γ levels. For each item, the first number indicates the compound number, and the second number indicates the concentration in nM; thus, "02 100" indicates that compound 2 was applied at a concentration of 100 nM, "04 200" indicates that compound 4 was applied at a concentration of 200 nM, and so on.

[0053] Figure 25(A) shows the evaluation of AhR transcripts in fibroblasts isolated from the intestinal mucosa of 6 normal patients (control group; CTR), 7 patients with ulcerative colitis (UC), and 7 patients with Crohn's disease (CD), normalized by real-time PCR and relative β-actin. Figure 25(B) shows flow cytometry analysis of AhR in fibroblasts isolated from 5 normal patients (CTR), 5 UC patients, and 5 CD patients. The right inset shows a representative histogram of aryl hydrocarbon receptor-expressing fibroblasts isolated from 1 CTR, 1 UC, and 1 CD patient. Staining with isotype control IgG is also shown. Data represent the mean + / - SD for all experiments. In summary, these data indicate constitutive AhR expression in intestinal fibroblasts.

[0054] Figure 26(A) shows the results of stimulation of Crohn's disease (CD) intestinal fibroblasts with TGF-β (1 ng / mL) in the presence or absence of Ficz (100, 200, or 400 nM). Figure 26(B) shows the results of stimulation of Crohn's disease (CD) intestinal fibroblasts with TNF-α (15 ng / mL) in the presence or absence of Ficz (100, 200, or 400 nM). Col1A1, Col3A1, and α-SMA were examined by real-time PCR 24 hours later. Data represent the mean + / - SD of three experiments. In summary, these data indicate that activation of AhR inhibits fibroblast collagen expression induced by pro-fibrotic cytokines.

[0055] Figure 27(A) shows the results of stimulation of Crohn's disease (CD) intestinal fibroblasts with TGF-β (1 ng / mL) in the presence or absence of CH223191 (a-AhR, final concentration 10 μM). Figure 27(B) shows the results of stimulation of Crohn's disease (CD) intestinal fibroblasts with TNF-α (15 ng / mL) in the presence or absence of CH223191 (a-AhR, final concentration 10 μM). Col1A1, Col3A1, and α-SMA were examined by real-time PCR 24 hours later. Data represent the mean + / - SD of three experiments.

[0056] Figures 28(A)-(D) show the total collagen analysis of the supernatant 48 hours after stimulation of Crohn's disease (CD) fibroblasts with TGF-β (1 ng / mL) ((A) and (C)) or TNF-α (15 ng / mL) ((B) and (D)) in the presence or absence of Ficz ((A) and (B)) or CH223191 (a-AhR, final concentration 10 μM) ((C) and (D)). Data represent the mean + / - SD of three experiments. In summary, these data indicate that AhR controls collagen secretion.

[0057] Figure 29(A) shows the percentage of CD intestinal fibroblasts expressing p-p38+, p-ERK1 / 2+, p-NF-κBp65+, or p-Smad2 / 3+ after 24 hours of unstimulation or stimulation with TGF-β (1 ng / mL) or TNF-α (15 ng / mL) in the presence or absence of Ficz (final concentration 200 nM) or CH223191 (a-AhR, final concentration 10 μM). Assessment of p-P38 (pT180 / pY182), p-ERK1 / 2 (pT202 / pY204)(pT184 / pY186), p-NF-κBp65, and p-Smad2 / 3 was performed by flow cytometry. The figures represent the percentage of p-p38+, p-ERK1 / 2+, p-NF-kBp65+, or p-Smad2 / 3+ cells in the specified phylum. Isotype control staining is also shown. One of three representative experiments using cells from three patients is shown. Figure 29(B) shows the percentage of p-p38+, p-ERK1 / 2+, p-NF-kBp65+, or p-Smad2 / 3+ fibroblasts isolated from three CD patients after 24 hours of stimulation with TGF-β (1 ng / mL) or TNF-α (15 ng / mL) in the presence or absence of Ficz (final concentration 200 nM) or CH223191 (a-AhR, final concentration 10 μM). Data represent the mean + / - SD of the three experiments. * p < 0.04 for unstimulated cells; ** p < 0.001 for unstimulated cells; # p < 0.03 for TGF-β; + p < 0.02 for TNF-α. In summary, these data demonstrate that AhR activation leads to the inactivation of p38 and ERK1 / 2 in Crohn's disease (CD) fibroblasts.

[0058] Figure 30(A) shows a schematic diagram of a TNBS-induced intestinal fibrosis model. Balb / c mice were treated with TNBS weekly, and Ficz or CH223191(a-AhR) was administered after 5 TNBS treatments. Figure 30(B) shows Masson's trichrome staining results of representative colonic cross sections from control (CTR) mice and TNBS-treated mice receiving Ficz or CH223191(a-AhR) at hour 12. The percentage of animals with mild, moderate, and severe fibrosis is also shown in the table. Figure 30(C) shows the relative RNA expression data of Col1A2 in colonic samples from CTR mice and TNBS-treated mice receiving Ficz or CH223191(a-AhR) injections, obtained by real-time PCR analysis. Figure 30(D) shows the total collagen content data (μg / mg) analyzed by colorimetric assay. Data represent the mean + / - SD of 3 independent experiments (n = 12 mice in each group). In summary, these data suggest that Ficz-treated mice are largely protected against TNBS-induced intestinal fibrosis. Invention Details

[0059] In one respect, this disclosure provides compounds according to formula I:

[0060]

[0061] Or its medication may accept salt, wherein:

[0062] A is

[0063] Ar by represent;

[0064] G 1 It is CR 4 Or N;

[0065] G 2 G 3 and G 4 Each is CR independently 4 2 or NR 1 ;

[0066] X 1 Each occurrence is independent of H, halogen, -OH, -CN, or C. 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 alkynyl group, C 3-6 cycloalkyl, C 1-6 Alkoxy, -O-phenyl, -N(R) 1 )2、-NO2、-C 1-6 Alkylene-N(R) 1 )2、-C(O)N(R3 )2、-CO2R 3 -C(O)R 3 -SR 3 -SO2R 3 -SO3R 3 or -SO2N(R) 3 )2;

[0067] X 2 Each occurrence is independent of H, halogen, -OH, -CN, or C. 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 alkynyl group, C 3-6 cycloalkyl, C 1-6 Alkoxy, -O-phenyl, -N(R) 1 )2、-NO2、-C 1-6 Alkylene-N(R) 1 )2、-C(O)N(R 3 )2、-CO2R 3 -SR 3 -SO2R 3 -SO3R 3 or -SO2N(R) 3 )2;

[0068] R 1 Each time it appears, it is independently either H or C. 1-6 alkyl;

[0069] R 2 Each time it appears, it is independently either H or C. 1-6 alkyl;

[0070] R 3 Each time it appears, it is independently selected from H and C. 1-6 Alkyl, phenyl, or heteroaryl;

[0071] R 4 Each time it appears, it is independently selected from H and C. 1-6 Alkyl and halogen;

[0072] Each occurrence of n is independently 0, 1, 2, or 3; and

[0073] Each time m appears, it is independently 0, 1, 2, 3, or 4;

[0074] Where C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 alkynyl group, C 3-6 cycloalkyl, C 1-6Each of the alkoxy or -O-phenyl groups may be optionally substituted by one, two or three substituents, each independently selected from halogen, -OH, -CN, -NR'R”-C(O)N(R'R”) and -C(O)R' (wherein R' and R” are each independently selected from H, methyl, ethyl, propyl or butyl, or R' and R” together form a 4-6 membered heterocycle);

[0075] Where A is At that time, G 1 It's CH, G 3 and G 4 It is CH2, and G 2 It is NR 1 X 2 It is not H (i.e., it does not exist, n is 0).

[0076] In some implementations, G 1 It is N.

[0077] In some implementations, G 2 It is NH.

[0078] In some implementations, G 1 It's CH, G 2 It is NH and G 3 and G 4 It is CH2.

[0079] In some implementations, G 1 It is N and G 2 G 3 and G 4 It is CH2.

[0080] In some implementations, G 1 It is N, G 2 and G 3 It is CH2 and G 4 It is NH.

[0081] In some implementations, G 1 It is N, G 2 and G 4 It is CH2 and G 3 It is NH.

[0082] In some implementations, G 1 It's CH, G 2 and G 4 It is NH and G 3 It is CH2.

[0083] In some implementations, Ar is And m is 1 or 2.

[0084] In some implementations, Ar is

[0085] In some implementations, X 1 It is H, halogen, -CN, or -OMe.

[0086] In some implementations, X 1 Yes-OMe.

[0087] In some implementations, X 2 Selected from H, halogens, -OH, -CN, C 1-6 Alkyl groups and CF3.

[0088] In some implementations, X 2 It is CF3 and n is 1.

[0089] In some embodiments, the compound is selected from...

[0090]

[0091] In one respect, this disclosure relates to compounds according to formula IA:

[0092]

[0093] Or its medication may accept salt, wherein:

[0094] X 1 If it exists, then each occurrence is C independently. 1-6 Alkyl, C 1-6 Alkoxy, -OC 1-6 alkylene-phenyl, C 1-6 Acyl group, -CO2R 3 -NO2, -OH or -N(R) 1 )2; or X 1 The two together can be -O(CH2)2O-;

[0095] X 2 Selected from H, C 1-6 Alkyl, C 1-6 Alkoxy, -OC 1-6 alkylene-phenyl and -OH;

[0096] R 1 Each time it appears, it is independently either H or C. 1-6 alkyl;

[0097] R 3 Each time it appears, it is independently H or C. 1-6 Alkyl, phenyl, or heteroaryl;

[0098] R 5 Selected from H, C1-6 Alkyl, C 1-6 Aryl and -CO2R 3 ;and

[0099] n is 0, 1, 2, or 3;

[0100] Where C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 alkynyl group, C 3-6 cycloalkyl, C 1-6 Each of the alkoxy and -O-phenyl groups is optionally substituted by one, two, or three substituents, each of which is independently selected from halogen, -OH, -CN, -NR'R”-C(O)N(R'R”) and -C(O)R' (wherein R' and R” are each independently selected from H, methyl, ethyl, propyl, or butyl, or R' and R” together form a 4-6 membered heterocycle);

[0101] In some implementation schemes, X 2 Selected from H, -OMe, ethyl, -OH and -OCH2Ph.

[0102] In some implementations, R 5 Selected from H, acetyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl and -CO2CH2Ph.

[0103] In some implementations, X 1 Each time it appears independently, it is H, -OMe, -NO2, -CO2Me, methyl, -OC(O)Me, -OCH2Ph, -OH, -NH2, or tert-butyl; or X 1 The two together can be -O(CH2)2O-.

[0104] In some implementations, n is 3.

[0105] In one respect, this disclosure relates to compounds according to formula II:

[0106]

[0107] Or its medication may accept salt, wherein:

[0108] B is

[0109] C is

[0110] Ar by represent;

[0111] X 1 If present, each occurrence is independently selected from halogen, -OH, -CN, C. 1-6Alkyl, C 2-6 Alkenyl, C 2-6 alkynyl group, C 3-6 cycloalkyl, C 1-6 Alkoxy, -O-phenyl, -N(R) 1 )2、-NO2、-C 1-6 Alkylene-N(R) 1 )2、-C(O)N(R 3 )2、-CO2R 3 -C(O)R 3 -SR 3 -SO2R 3 -SO3R 3 or -SO2N(R) 3 )2;

[0112] X 2 If present, each occurrence is independently selected from halogen, -OH, -CN, C. 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 alkynyl group, C 3-6 cycloalkyl, C 1-6 Alkoxy, -O-phenyl, -N(R) 1 )2、-NO2、-C 1-6 Alkylene-N(R) 1 )2、-C(O)N(R 3 )2、-CO2R 3 -SR 3 -SO2R 3 -SO3R 3 or -SO2N(R) 3 )2;

[0113] R 1 Each time it appears, it is independently either H or C. 1-6 alkyl;

[0114] R 3 Each time it appears, it is independently H or C. 1-6 Alkyl, phenyl, or heteroaryl;

[0115] R 4 Each time it appears, it is independently selected from H and C. 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 alkynyl group, -N(R) 1 )2、-C 1-6 Alkylene-N(R) 1 )2、-C(O)N(R 3 )2 and -CO2R 3 ;

[0116] Each time n appears, it is independently 0, 1, 2, or 3;

[0117] Each time m appears, it is independently 0, 1, 2, 3, or 4;

[0118] Where C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 alkynyl group, C 3-6 cycloalkyl, C 1-6 Each of the alkoxy and -O-phenyl groups is optionally substituted by one, two, or three substituents, each of which is independently selected from halogen, -OH, -CN, -NR'R”-C(O)N(R'R”) and -C(O)R' (wherein R' and R” are each independently selected from H, methyl, ethyl, propyl, or butyl, or R' and R” together form a 4-6 membered heterocycle);

[0119] X 3 Is it N or CR? 4 ;

[0120] X 4 It is NR 1 , O or S; and

[0121] Y represents a chemical bond, C 1-6 Alkylene or -N(R) 1 )-;

[0122] Where B is And X 4 When it is S, X 2 It is not H (i.e., it does not exist).

[0123] In some embodiments of Formula II, Y is CH2 or NH.

[0124] In some implementations, R 4 Selected from C 2-6 alkenyl, C 2-6 alkynyl group, -C 1-6 Alkylene-N(R) 1 )2 and -C(O)N(R 3 )2.

[0125] In some implementations, R 4 Is it allyl or

[0126] In some implementations, Ar is And m is 1 or 2.

[0127] In some implementations, Ar is

[0128] In some implementations, X 1It is H (i.e., n is 0), halogen, -CN, or -OMe.

[0129] In some embodiments, A, B, and L together represent compounds as follows:

[0130]

[0131] In some implementations, X 2 Selected from H, halogens, -OH, -CN, C 1-6 Alkyl groups and CF3.

[0132] In some implementations, X 2 It is CF3 and n is 1.

[0133] In one respect, this disclosure relates to compounds according to formula III:

[0134]

[0135] Or its medication may accept salt, in which

[0136] D is

[0137] Ar by represent;

[0138] X 1 It can exist, and each time it appears, it is independently selected from halogen, -OH, -CN, C. 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 alkynyl group, C 3-6 cycloalkyl, C 1-6 Alkoxy, -O-phenyl, -N(R) 1 )2、-NO2、-C 1-6 Alkylene-N(R) 1 )2、-C(O)N(R 3 )2、-CO2R 3 -C(O)R 3 -SR 3 -SO2R 3 -SO3R 3 or -SO2N(R) 3 )2;

[0139] X 2 It can exist, and each time it appears, it is independently selected from halogen, -OH, -CN, C. 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 alkynyl group, C 3-6 cycloalkyl, C 1-6 Alkoxy, -O-phenyl, -N(R)1 )2、-NO2、-C 1-6 Alkylene-N(R) 1 )2、-C(O)N(R 3 )2、-CO2R 3 -SR 3 -SO2R 3 -SO3R 3 or -SO2N(R) 3 )2;

[0140] R 1 Each time it appears, it is independently either H or C. 1-6 alkyl;

[0141] R 3 Each time it appears, it is independently H or C. 1-6 Alkyl, phenyl, or heteroaryl;

[0142] Each occurrence of n is independently 0, 1, 2, or 3; and

[0143] Each time m appears, it is independently 0, 1, 2, 3, or 4;

[0144] Where C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 alkynyl group, C 3-6 cycloalkyl, C 1-6 Each of the alkoxy and -O-phenyl groups is optionally substituted by one, two, or three substituents, each of which is independently selected from halogen, -OH, -CN, -NR'R”-C(O)N(R'R”) and -C(O)R' (wherein R' and R” are each independently selected from H, methyl, ethyl, propyl, or butyl, or R' and R” together form a 4-6 membered heterocycle);

[0145] Where D is At that time, X 2 Not H (i.e., X) 2 (Does not exist and n is 0).

[0146] In some implementations, Ar is And m is 1 or 2.

[0147] In some implementations, Ar is

[0148] In some implementations, X 1 It is H, halogen, -CN, or -OMe.

[0149] In some implementations, X 1 Yes-OMe.

[0150] In some implementations, X 2 Selected from H, halogens, -OH, -CN, C 1-6 Alkyl groups and CF3.

[0151] In some implementations, X 2 It is CF3 and n is 1.

[0152] In one respect, this disclosure relates to compounds according to formula IV:

[0153]

[0154] Or its medication may accept salt, wherein:

[0155] E is selected from heterocyclic groups, 5-membered heteroaryl groups, 6-membered heteroaryl groups, aryl groups, and C. 3-6 Cycloalkyl rings, wherein the rings are optionally composed of 1, 2 or 3 X's 2 Replaced;

[0156] Ar by represent;

[0157] X 1 If present, each occurrence is independently selected from H, halogen, -OH, -CN, C. 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 alkynyl group, C 3-6 cycloalkyl, C 1-6 Alkoxy, -O-phenyl, -N(R) 1 )2、-NO2、-C 1-6 Alkylene-N(R) 1 )2、-C(O)N(R 3 )2、-CO2R 3 -C(O)R 3 -SR 3 -SO2R 3 -SO3R 3 or -SO2N(R) 3 )2;

[0158] X 2 Each occurrence is independently selected from halogens, -OH, -CN, and C. 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 alkynyl group, C 3-6 cycloalkyl, C 1-6 Alkoxy, -O-phenyl, -N(R) 1 )2、-NO2、-C 1-6 Alkylene-N(R) 1 )2、-C(O)N(R 3 )2、-CO2R3 -SR 3 -SO2R 3 -SO3R 3 or -SO2N(R) 3 )2.

[0159] Y is selected from chemical bonds, NR', or C. 1-6 Alkylene (e.g., chemical bond or C) 1-2 Alkylene);

[0160] R 1 Each time it appears, it is independently either H or C. 1-6 alkyl;

[0161] R 3 Each time it appears, it is independently selected from H and C. 1-6 Alkyl, phenyl, or heteroaryl; and

[0162] Each time m appears, it is independently 0, 1, 2, 3, or 4;

[0163] Where C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 alkynyl group, C 3-6 cycloalkyl, C 1-6 Each of the alkoxy and -O-phenyl groups is optionally substituted by one, two, or three substituents, each of which is independently selected from halogen, -OH, -CN, -NR'R”-C(O)N(R'R”) and -C(O)R' (wherein R' and R” are each independently selected from H, methyl, ethyl, propyl, or butyl, or R' and R” together form a 4-6 membered heterocycle);

[0164] In some embodiments of Formula IV, Y is a chemical bond, CH2 or NH; (e.g., Y can be a chemical bond).

[0165] In some implementations, Ar is And m is 1 or 2.

[0166] In some implementations, Ar is

[0167] In some implementations, X 1 It is H, halogen, -CN, or -OMe.

[0168] In some implementations, X 1 Yes-OMe.

[0169] In some implementations, X 2 Selected from H, halogens, -OH, -CN, C 1-6 Alkyl groups and CF3.

[0170] In some implementations, X 2 It's CF3.

[0171] In one respect, this disclosure relates to pharmaceutical compositions comprising the aforementioned compounds; and pharmaceutically acceptable carriers.

[0172] definition

[0173] As used herein, the term "saturated" means that a compound is modified to have neither carbon-carbon double bonds nor carbon-carbon triple bonds, except as otherwise noted below. This term does not exclude carbon-heteroatom multiple bonds, such as carbon-oxygen double bonds or carbon-nitrogen double bonds. Furthermore, it does not exclude the presence of carbon-carbon double bonds in moieties such as keto-enol tautomers or imine-enamine tautomers.

[0174] The term "alkyl" refers to a monovalent saturated aliphatic group having a carbon atom as a linker, a straight or branched chain, a cyclic or acyclic structure, and no atoms other than carbon and hydrogen. Therefore, cycloalkyl, as used herein, is a subset of alkyl groups. The groups -CH3(Me), -CH2CH3(Et), -CH2CH2CH3(n-Pr), -CH(CH3)2(i-Pr), -CH(CH2)2(cyclopropyl), -CH2CH2CH2CH3(n-Bu), -CH(CH3)CH2CH3(sec-butyl), -CH2CH(CH3)2(isobutyl), -C(CH3)3(tert-butyl), -CH2C(CH3)3(neopentyl), cyclobutyl, cyclopentyl, cyclohexyl, and cyclohexylmethyl are non-limiting examples of alkyl groups. The term "alkylene" refers to a divalent saturated aliphatic group with one or two saturated carbon atoms as bonding points, a straight or branched chain, a cyclic, cyclic, or acyclic structure, lacking carbon-carbon double or triple bonds, and containing no atoms other than carbon and hydrogen. Examples of such groups include -CH2- (methylene), -CH2CH2-, -CH2C(CH3)2CH2-, and -CH2CH2CH2-. These are non-limiting examples of alkylene groups.

[0175] The term "alkenyl" refers to a monovalent unsaturated aliphatic group having a carbon atom as a linker, a straight or branched chain, a cyclic or acyclic structure, at least one non-aromatic carbon-carbon double bond, no carbon-carbon triple bonds, and no atoms other than carbon and hydrogen. Non-limiting examples of alkenyl groups include: -CH═CH2 (vinyl), -CH═CHCH3, -CH═CHCH2CH3, -CH2CH═CH2 (allyl), -CH2CH═CHCH3, and -CH═CH-C6H5.

[0176] The term "alkynyl" refers to a monovalent unsaturated aliphatic group having a carbon atom as a linker, a straight or branched chain, a cyclic or acyclic structure, at least one non-aromatic carbon-carbon triple bond, and no atoms other than carbon and hydrogen. As used herein, the term alkynyl does not exclude the presence of one or more non-aromatic carbon-carbon double bonds. Non-limiting examples of alkynyl groups include the groups -C≡CH, -C≡CCH3, and -CH2C≡CCH3.

[0177] The term "aryl" refers to a monovalent group having an aromatic carbon atom as a connecting point, the carbon atom forming one or more six-membered aromatic ring structures, wherein all ring atoms are carbon, and the monovalent group contains no atoms other than carbon and hydrogen. Non-limiting examples of aryl groups include phenyl (Ph), benzyl, (dimethyl)phenyl, -C6H4-CH2CH3 (phenylethyl), -C6H4-CH2CH2CH3 (phenylpropyl), -C6H4-CH(CH3)2, -C6H4-CH(CH2)2, -C6H3(CH3)CH2CH3 (methylethylphenyl), -C6H4-CH═CH2 (vinylphenyl), -C6H4-CH═CHCH3, -C6H4C≡CH, -C6H4C≡CCH3, naphthyl, and monovalent groups derived from biphenyl.

[0178] The term "heteroaryl" refers to a monovalent aromatic group having an aromatic carbon or nitrogen atom as a connecting point, the carbon or nitrogen atom forming part of one or more aromatic ring structures, wherein at least one ring atom is nitrogen, oxygen, or sulfur, and wherein the heteroaryl group does not contain any atoms other than carbon, hydrogen, aromatic nitrogen, aromatic oxygen, and aromatic sulfur. As used herein, this term does not exclude the presence of one or more alkyl, aryl, and / or aralkyl groups (where the carbon number limit allows) connected to the aromatic ring or aromatic ring system. If more than one ring is present, these rings may be fused or non-fused. Non-limiting examples of heteroaryl groups include furanyl, imidazolyl, indolyl, indazole, isoxazolyl, methylpyridinyl, oxazolyl, phenylpyridinyl, pyridinyl, pyrroleyl, pyrimidinyl, pyrazinyl, quinolinyl, quinazolinyl, quinoxalinyl, triazinyl, tetrazolyl, thiazolyl, thiopheneyl, and triazolyl.

[0179] The term "heterocyclic group" refers to a monovalent non-aromatic group having a carbon or nitrogen atom as a connecting point, wherein the carbon or nitrogen atom forms part of one or more non-aromatic ring structures, wherein at least one ring atom is nitrogen, oxygen, or sulfur, and wherein the heterocyclic group contains no atoms other than carbon, hydrogen, nitrogen, oxygen, and sulfur. As used herein, the term does not exclude the presence of one or more alkyl groups (where the carbon number limit allows) attached to the ring or ring system. If more than one ring is present, these rings may be fused or unfused. Non-limiting examples of heterocyclic groups include azirropropyl, pyrrolidinyl, piperidinyl, piperazine, morpholinyl, thiomorpholinyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, and pyranyl.

[0180] The term "acyl" refers to the group -C(O)R, where R is hydrogen, alkyl, aryl, aralkyl, or heteroaryl, as defined above. The groups -CHO, -C(O)CH3 (acetyl, Ac), -C(O)CH2CH3, -C(O)CH2CH2CH3, -C(O)CH(CH3)2, -C(O)CH(CH2)2, -C(O)C6H5, -C(O)C6H4-CH3, -C(O)CH2C6H5, and -C(O) (imidazolyl) are non-limiting examples of acyl groups. "Thioacyl" is defined similarly, except that the oxygen atom in the -C(O)R group is replaced by a sulfur atom.

[0181] The term "alkoxy" refers to the group -OR, where R is an alkyl group, as defined above. Non-limiting examples of alkoxy groups include: -OCH3, -OCH2CH3, -OCH2CH2CH3, -OCH(CH3)2, -OCH(CH2)2, -O-cyclopentyl, and -O-cyclohexyl.

[0182] The definition of "drug-acceptable" refers to any carrier, salt form, or other reagent that does not interfere with the efficacy of the biological activity of the active ingredient and is non-toxic to the host to which it is administered. As used herein, "drug-acceptable carrier" means a buffer, carrier, diluent, or excipient suitable for contact with human and animal tissues without producing excessive toxicity, radioactivity, immune response, or other problems or complications, while having a reasonable benefit / risk ratio. The carrier should be "acceptable" in the sense of compatibility with other components of the formulation and should not be harmful to the recipient. Pharmaceutically acceptable carriers include buffers, solvents, dispersion media, coatings, isotonic agents, and absorption delay agents compatible with drug administration. "Drug-acceptable salt" refers to a salt of the compounds of this disclosure that is pharmaceutically acceptable and has the desired pharmaceutical activity. Such salts include acid addition salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; or acid addition salts formed with organic acids, such as 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, 2-naphthalenesulfonic acid, 3-phenylpropionic acid, 4,4'-methylenebis(3-hydroxy-2-en-1-carboxylic acid), 4-methylbicyclo[2.2.2]oct-2-en-1-carboxylic acid, acetic acid, aliphatic monocarboxylic acids and dicarboxylic acids, aliphatic sulfuric acid, aromatic sulfuric acid, benzenesulfonic acid, benzoic acid, etc. Camphor sulfonic acid, carbonic acid, cinnamic acid, citric acid, cyclopentanepropionic acid, ethanesulfonic acid, fumaric acid, glucoheponic acid, gluconic acid, glutamic acid, glycolic acid, heptanoic acid, hexanoic acid, hydroxynaphthenic acid, lactic acid, lauryl sulfate, maleic acid, malic acid, malonic acid, mandelic acid, mesylic acid, mucoconic acid, o-(4-hydroxybenzoyl)benzoic acid, oxalic acid, p-chlorobenzenesulfonic acid, phenyl-substituted alkanic acids, propionic acid, p-toluenesulfonic acid, pyruvic acid, salicylic acid, stearic acid, succinic acid, tartaric acid, tert-butylacetic acid, trimethylacetic acid, etc. Acceptable drug salts also include base addition salts that can be formed when the present acidic protons can react with inorganic or organic bases. Acceptable inorganic bases include sodium hydroxide, sodium carbonate, potassium hydroxide, aluminum hydroxide, and calcium hydroxide. Acceptable organic bases include ethanolamine, diethanolamine, triethanolamine, aminobutanetriol, N-methylglucosamine, etc. It should be recognized that the specific anion or cation forming any part of the salt of the present invention is not critical, as long as the salt as a whole is pharmaceutically acceptable. Other examples of pharmaceutically acceptable salts, their preparation and use are given in the Handbook of Pharmaceutical Salts Properties, and Use (PHStahl & C.G. Wermutheds., Verlag Helvetica Chimica Acta, 2002), which is incorporated herein by reference in its entirety.

[0183] The compounds disclosed herein may contain one or more chiral centers and thus exist as stereoisomers. As used herein, the term "stereoisomer" comprises all enantiomers or diastereomers. Depending on the configuration of the substituents surrounding the stereocarbon atom, these compounds may be designated by the symbols "(+)", "(-)", "R", or "S", but those skilled in the art will recognize that certain structures implicitly represent a chiral center. The present invention includes a variety of stereoisomers of these compounds and mixtures thereof. Mixtures of enantiomers or diastereomers may be designated as "(±)" in the nomenclature, but those skilled in the art will recognize that certain structures implicitly represent a chiral center.

[0184] The compounds disclosed herein may contain one or more double bonds and thus exist as geometric isomers resulting from the arrangement of substituents around the carbon-carbon double bonds. (Symbols) The bonds described herein can be single, double, or triple bonds. Substituents surrounding the carbon-carbon double bond are designated as being in the "Z" or "E" configuration, where the terms "Z" and "E" are used according to IUPAC standards. Unless otherwise stated, the structure of the double bond is described including both "E" and "Z" isomers. Substituents around the carbon-carbon double bond can be referred to as "cis" or "trans," where "cis" indicates a substituent on the same side of the double bond, and "trans" indicates a substituent on the opposite side of the double bond.

[0185] The compounds disclosed herein may contain a carbocyclic or heterocyclic ring and thus exist as geometric isomers resulting from the arrangement of substituents around the ring. The arrangement of substituents around the carbocyclic or heterocyclic ring is designated as a “Z” or “E” configuration, wherein the terms “Z” and “E” are used according to IUPAC standards. Unless otherwise stated, the description of the structure of the carbocyclic or heterocyclic ring includes both “Z” and “E” isomers. Substituents around the carbocyclic or heterocyclic ring may also be referred to as “cis” or “trans”, wherein the term “cis” indicates a substituent on the same side of the ring plane, and the term “trans” indicates a substituent on the opposite side of the ring plane. A mixture of compounds in which substituents are located on the same and opposite sides of the ring plane is called a “cis / trans” compound.

[0186] Individual enantiomers and diastereomers of the compounds of the present invention can be synthesized from commercially available starting materials containing asymmetric or stereocenters, or prepared by preparing a racemic mixture and then by analytical methods well known to those skilled in the art. Examples of such analytical methods include: (1) attaching a mixture of enantiomers to a chiral auxiliary, separating the resulting diastereomer mixture by recrystallization or chromatography, and releasing an optically pure product from the auxiliary; (2) salt formation using an optically active analytical agent; (3) direct separation of the optically enantiomer mixture on a chiral liquid chromatography column; or (4) kinetic analysis using stereoselective chemical or enzymatic reagents. Racemic mixtures can also be resolved to their component enantiomers by well-known methods, such as by chiral liquid chromatography or by crystallizing the compound in a chiral solvent. Stereoselective synthesis is a chemical or enzymatic reaction in which a single reactant forms an unequal mixture of stereoisomers during the generation of a new stereocenter or during the transformation of a previously existing stereocenter. Stereoselective synthesis includes enantioselective and diastereoselective conversions and may involve the use of chiral auxiliaries. For example, see Carreira and Kvaerno, Classics in Stereoselective Synthesis, Wiley-VCH:Weinheim, 2009.

[0187] The compounds disclosed herein can exist in both solvated and non-solvated forms with pharmaceutically acceptable solvents such as water, ethanol, etc., and the invention is intended to include both solvated and non-solvated forms. In one embodiment, the compound is amorphous. In one embodiment, the compound is a single polymorph. In another embodiment, the compound is a mixture of polymorphs. In yet another embodiment, the compound is in crystalline form.

[0188] This invention also includes isotopically labeled compounds of the invention, which are identical to those described herein, except that one or more atoms are replaced by atoms with atomic masses or mass numbers different from those normally found in nature. Examples of isotopes that may be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, respectively, for example... 2 H, 3 H, 13 C 14 C 15 N、 18 O、 17 O、 31 P, 32 P, 35 S, 18 F and 36 Cl. For example, the compounds of the present invention may have one or more deuterium-substituted H atoms.

[0189] Disclosed compounds labeled with certain isotopes (e.g., using...) 3 H and 14 Those labeled with C can be used for the determination of the tissue distribution of compounds and / or substrates. Tritium (i.e., 3 H) and carbon-14 (i.e. 14 C) Isotopes are particularly preferred because they are easy to prepare and detect. Furthermore, heavier isotopes such as deuterium (i.e., 2 Replacing non-isotopically labeled reagents with isotopically labeled reagents can provide greater metabolic stability (e.g., increased in vivo half-life or reduced dose requirements) and thus may be preferred in some cases. The isotopically labeled compounds of the present invention can generally be prepared by replacing non-isotopically labeled reagents with isotopically labeled reagents using methods similar to those disclosed in the embodiments herein.

[0190] As used herein, “patient” or “subject” means any animal at risk, suffering from, or diagnosed with an inflammatory condition, including but not limited to mammals, primates, and humans. In some embodiments, a patient may be a non-human mammal, such as a cat, dog, or horse. A patient may be an individual diagnosed as being at high risk of developing an inflammatory condition, an individual already diagnosed with an inflammatory condition, an individual with a prior history of an inflammatory condition, or an individual assessed as having symptoms or indications for an inflammatory condition.

[0191] As used herein, "patient in need" means a patient who has any symptoms or manifestations of an inflammatory condition, a patient who may have any symptoms or manifestations of an inflammatory condition, or a patient who may benefit from the method of the present invention for treating an inflammatory condition, such as inflammatory bowel disease. Patients in need may include patients diagnosed as being at risk of developing an inflammatory condition (e.g., inflammatory bowel disease), patients who already have an inflammatory condition, or patients who have previously received treatment for such a condition.

[0192] As used herein, the terms “treatment,” “management,” “disposition,” etc., generally refer to achieving the desired pharmaceutical and / or physiological effect. Such effect may be preventative in terms of complete or partial prevention of the disease or its symptoms, and / or therapeutic in terms of partial or complete cure of the disease and / or adverse effects attributable to the disease. The term “treatment” as used herein encompasses any treatment of a disease in mammals, particularly humans, including: (a) prevention of the onset of the disease in subjects who may be susceptible to the disease but have not yet been diagnosed with it; (b) suppression of the disease, i.e., prevention of an increase in the severity or extent of the disease; (c) relief of the disease, i.e., achieving partial or complete improvement of the disease; or (d) prevention of relapse of the disease, i.e., prevention of the disease from returning to an active state following previous successful treatment of the disease symptoms or treatment of the disease.

[0193] As used herein, "effective amount" refers to an amount of agent sufficient to at least partially treat a condition when administered to a patient. Therapeutic effective amounts will vary depending on the severity of the condition, the route of administration of the component, and the age, weight, etc., of the patient being treated. Therefore, the effective amount of a compound disclosed herein is the amount of such compound required in a patient to treat one or more conditions or diseases considered herein, such that administration of the drug prevents the occurrence of the condition in the subject, prevents the progression of the condition (e.g., prevents the onset or increase in the severity of symptoms), or alleviates or completely improves all associated symptoms of the condition, even to the point of remission. An effective amount can also be the amount of such compound necessary to produce the desired biological effect, for example, lowering INF-γ levels.

[0194] The efficacy of treatment can be evaluated by assessing overall symptoms associated with the inflammatory condition, histological analysis of tissues, biochemical assays, imaging methods such as magnetic resonance imaging, or other known methods. For example, the efficacy of treatment can be assessed by analyzing overall symptoms of the condition following administration of the compounds described herein, such as changes in tissue inflammation, abdominal pain, vomiting, diarrhea, rectal bleeding, cramps, muscle spasms, weight loss, malnutrition, fever, anemia, or other overall pathological aspects associated with the inflammatory condition.

[0195] Treatment efficacy can also be evaluated at the tissue or cellular level, for example, by obtaining tissue biopsy specimens (e.g., gastrointestinal tissue biopsy specimens) and assessing overall tissue or cell morphology or staining characteristics. Biochemical assays examining protein or RNA expression can also be used to evaluate treatment efficacy. For example, the levels of IL-22, IFN-γ, or other proteins or gene products indicating one or more inflammatory conditions, inflammatory cytokine production, or IL-22-mediated inflammatory responses in dissociated cells or non-dissociated tissues can be evaluated using immunocytochemistry, immunohistochemistry, Western blotting or Northern blotting methods, or methods for assessing RNA levels such as quantitative or semi-quantitative polymerase chain reaction. The presence or expression levels of useful biomarkers found in feces, plasma, or serum can also be assessed to evaluate disease status and treatment efficacy.

[0196] method

[0197] This disclosure provides methods for treating inflammatory conditions in patients in need, including administering an effective amount of the compounds disclosed herein to the patient. Exemplary inflammatory conditions considered herein include Crohn's disease, gastroduodenal Crohn's disease, Crohn's disease (granulomatous) colitis, ulcerative colitis, collagenous colitis, lymphocytic colitis, ischemic colitis, metastatic colitis, Behcet's disease, microscopic colitis, ulcerative proctitis, rectal colitis, jejunal bend, left colitis, panenteritis, ileocolic colitis, ileitis, indeterminate colitis, and eosinophilic colitis. Other anticipated inflammatory conditions include cartilage inflammation, bone degeneration, rheumatoid arthritis, arthritis, psoriatic arthritis, allergic pneumonia, liver disease (such as fatty liver, hepatitis, hepatic steatosis, and non-alcoholic steatohepatitis (NASH)), fibrosis (e.g., intestinal fibrosis, pulmonary fibrosis, or liver fibrosis), autoimmune polyendocrine syndrome, Addison's disease, Goodpassch's syndrome, Graves' disease, Guillain-Barré syndrome, Hashimoto's encephalopathy, psoriasis, Hashimoto's thyroiditis, juvenile arthritis (e.g., juvenile idiopathic arthritis), lupus erythematosus, multiple sclerosis, Alzheimer's disease, dermatitis, atopic dermatitis, acne, type I diabetes, autoimmune hepatitis, Meniere's disease, Raynaud's phenomenon, Sjögren's syndrome, ankylosing spondylitis, chronic fatigue syndrome, polymyalgia rheumatica, osteoarthritis, prostatitis, and Reiter's syndrome. In some implementations, the patient is a mammal. In some other implementations, the patient is a human.

[0198] In some embodiments, a method for treating an inflammatory disease or condition is provided, wherein said disease or condition is selected from inflammatory bowel disease, chondritis, bone degeneration, ulcerative colitis, psoriasis, arthritis, psoriatic arthritis, rheumatoid arthritis, juvenile arthritis, juvenile rheumatoid arthritis, autoimmune hepatitis, Crohn's disease, lupus erythematosus, multiple sclerosis, Alzheimer's disease, dermatitis, atopic dermatitis, acne, type 1 diabetes mellitus, Raynaud's phenomenon, Graves' disease, and Addison's disease. And said method includes administering an effective amount of the disclosed compound to a patient in need. In some embodiments, the patient is a mammal. In some other embodiments, the patient is a human.

[0199] Methods for treating Crohn's disease and / or ulcerative colitis and / or inflammatory bowel disease are provided, comprising administering an effective amount of the disclosed compound to a patient in need. In some embodiments, the patient is a mammal. In some other embodiments, the patient is a human.

[0200] In some implementations, the inflammatory disease or condition described is selected from Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, metastatic colitis, Behcet's disease, idiopathic inflammatory bowel disease, irritable bowel syndrome, regional enteritis, spastic colon, microscopic colitis, Crohn's colitis, perianal disease, and indeterminate colitis.

[0201] In some embodiments, the inflammatory disease or condition is Crohn's disease. In some embodiments, Crohn's disease is selected from ileocolitis, ileitis, gastroduodenal Crohn's disease, jejunoileitis, and granulomatous ileocolitis. In some embodiments, Crohn's disease includes intestinal fibrosis. In some embodiments, Crohn's disease is fibrotic stricture Crohn's disease.

[0202] On the other hand, this disclosure provides methods for preventing, treating, or alleviating fibrotic stenosis or intestinal fibrosis in subjects, including administering a therapeutically effective amount of the disclosed compound to a subject in need. In some embodiments, the fibrotic stenosis or intestinal fibrosis is associated with Crohn's disease.

[0203] It is understood that inflammatory bowel disease can be associated with a number of symptoms. Therefore, this disclosure provides a method for relieving one or more symptoms of inflammatory bowel disease, comprising administering an effective amount of the disclosed compound to a patient in need, the symptoms being selected from abdominal pain, vomiting, diarrhea, rectal bleeding, severe cramps, muscle spasms, weight loss, malnutrition, fever, anemia, skin lesions, joint pain, eye inflammation, liver disease, arthritis, pyoderma gangrenosa, primary sclerosing cholangitis, nonthyroidal syndromes, and growth defects in children.

[0204] In one aspect, this disclosure relates to methods for reducing IFN-γ levels and / or inhibiting IFN-γ, including administering a therapeutically effective amount of the disclosed compound to a subject in need. As used herein, “inhibition of IFN-γ” can refer to a complete or partial reduction in IFN-γ expression or activity. Therefore, inhibition of IFN-γ can refer to an alteration in the state of the IFN-γ gene or chromatin, or an alteration in the interaction with regulators of gene transcription or gene accessibility, resulting in a complete or partial reduction in the expression of IFN-γ gene products such as IFN-γ RNA, IFN-γ protein, or IFN-γ peptide sequences. Inhibition of IFN-γ can also refer to the inhibition of processes essential to the expression of IFN-γ gene products, including but not limited to IFN-γ transcription, IFN-γ RNA processing, IFN-γ protein translation, or post-translational modification of IFN-γ. Additionally, inhibition of IFN-γ can refer to the inhibition of the activity of IFN-γ gene products (including IFN-γ peptides, IFN-γ nucleotide products (e.g., IFN-γ mRNA), and IFN-γ proteins). Inhibition of IFN-γ gene product activity can include a reduction in IFN-γ signaling or a reduction in the direct or indirect interactions of IFN-γ with other cellular components (e.g., proteins, peptides, DNA, RNA, lipids, or signaling molecules), including the nucleus, organelles, cytosol, membranes, and extracellular components. For example, inhibition of IFN-γ activity can include inhibiting IFN-γ binding to or activation of CSF1R or inhibiting downstream signaling effects of CSF1R (e.g., MAP kinase phosphorylation or macrophage proliferation).

[0205] In one aspect, this disclosure relates to a method for increasing IL-22 levels, comprising administering a therapeutically effective amount of the disclosed compound to a subject in need. This increase can be used to enhance or strengthen immunity or provide immune protection.

[0206] This invention also provides a method for increasing IL-22 production in the cells of patients with inflammatory conditions, comprising administering an effective amount of the disclosed compound to the patient in need. IL-22 production can be increased in any cells in which IL-22 expression or activity occurs, including cells in the gastrointestinal tract, immune system, and blood. Cells of the gastrointestinal tract (including cells of the stomach, duodenum, jejunum, ileum, colon, rectum, and anal canal) include columnar epithelial cells, mucosal epithelial cells, zymogen cells, neck mucus cells, parietal cells, gastrinocytes, goblet cells, paneth cells, oligomucosa cells, and villous absorptive cells. Cells of the immune system include leukocytes, phagocytes (e.g., macrophages, neutrophils, and dendritic cells), monocytes, mast cells, eosinophils, basophils, natural killer cells, innate cells, lymphocytes, B cells, and T cells. Blood cells include red blood cells (erythrocytes) and white blood cells (leukocytes, monocytes, and platelets).

[0207] In one aspect, this disclosure relates to methods for increasing IL-22 levels and decreasing IFN-γ levels, comprising administering a therapeutically effective amount of the aforementioned compound to a subject in need.

[0208] In one aspect, this disclosure relates to a method of inhibiting lipid peroxidation, comprising administering a therapeutically effective amount of the aforementioned compound to a subject in need.

[0209] In one aspect, this disclosure relates to a method of modulating an aryl hydrocarbon receptor (AhR), comprising administering a selective AhR modulator; wherein the selective AhR modulator is the aforementioned compound.

[0210] In another aspect, the present invention provides compounds as described herein for use as medicines. In another aspect, the present invention provides methods for using compounds as described herein for treating inflammatory diseases or conditions. Inflammatory diseases or conditions can be as defined above. In another aspect, the present invention provides methods for using compounds as described herein for treating Crohn's disease and / or ulcerative colitis and / or inflammatory bowel disease. Crohn's disease can be as defined above. In another aspect, this disclosure provides methods for using compounds as described herein for preventing, treating, or reducing fibrotic strictures or intestinal fibrosis. In some embodiments, fibrotic strictures or intestinal fibrosis are associated with Crohn's disease. In another embodiment, the present invention provides methods for using compounds as described herein to relieve one or more symptoms of inflammatory bowel disease. The symptoms can be as defined above. In one aspect, the present invention provides compounds as described herein relating to methods for reducing IFN-γ levels and / or inhibiting IFN-γ in subjects of need. Inhibition of IFN-γ can be as defined above. In another aspect, the present invention provides methods for using compounds as described herein to increase IL-22 levels in an individual. The increase can be used to enhance or strengthen immunity or provide immune protection. The present invention also provides a method for using the compounds described above to increase IL-22 production in cells of patients with inflammatory conditions. Inflammatory conditions can be as defined above. In another aspect, the present invention provides a method for using the compounds described herein to increase IL-22 levels and decrease IFN-γ levels in an individual. The present invention also provides a method for using the compounds described herein to inhibit lipid peroxidation in a subject. The present invention further provides a method for using the compounds to regulate aryl hydrocarbon receptors (AhR) in a subject.

[0211] Pharmaceutical Compositions and Routes of Administration

[0212] The present invention also provides pharmaceutical compositions comprising the compounds described herein. In another aspect, the present invention provides pharmaceutical compositions for treating inflammatory conditions. These pharmaceutical compositions may comprise the compounds described herein and a drug-acceptable carrier. As used herein, the term "pharmaceutical composition" refers, for example, a mixture containing a specified amount of a therapeutic compound, such as a therapeutically effective amount of a therapeutic compound mixed with a drug-acceptable carrier, to be administered to a mammal, such as a human, to treat an inflammatory condition.

[0213] The compounds disclosed herein can be specifically formulated for administration to a subject in solid, liquid, or gel form, including forms suitable for: (1) oral administration, such as oral enemas (aqueous or non-aqueous solutions or suspensions), lozenges, sugar-coated pills, capsules, pellets, tablets (e.g., those for oral, sublingual, and systemic absorption), boluses, powders, granules, pastes for application to the tongue; (2) parenteral administration, such as by subcutaneous, intramuscular, intravenous, or epidural injection, such as sterile solutions or suspensions, or sustained-release formulations; (3) topical administration, such as as creams, ointments, controlled-release patches, or sprays applied to the skin; (4) intravaginal or rectal administration, such as as vaginal suppositories, creams, or foams; (5) sublingual administration; (6) ocular administration; (7) transdermal administration; (8) transmucosal administration; or (9) nasal administration. Examples of dosage forms include, but are not limited to: tablets; capsule tablets; capsules, such as hard gelatin capsules and soft elastic gelatin capsules; flat capsules; lozenges; tablets; dispersants; suppositories; ointments; poultices (mud dressings); pastes; powders; dressings; creams; plasters; solutions; patches; aerosols (e.g., nasal sprays or inhalers); gels; liquids such as suspensions (e.g., aqueous or non-aqueous liquid suspensions, oil-in-water emulsions, or water-in-oil emulsions), solutions, and elixirs; and sterile solids (e.g., crystalline or amorphous solids) that can be reconfigured to provide liquid dosage forms.

[0214] Pharmaceutical compositions containing the compounds described herein may be available in dosage units and may be prepared by any suitable method. The pharmaceutical composition shall be formulated to be compatible with its intended route of administration. Available formulations may be prepared by methods well known in the pharmaceutical industry. See, for example, Remington's Pharmaceutical Sciences, 18th ed. (Mack Publishing Company, 1990).

[0215] The pharmaceutical formulation is preferably sterile. Sterilization can be achieved, for example, by filtration through a sterile filter membrane. When the composition is in lyophilized form, filtration sterilization can be performed before or after lyophilization and reconstitution.

[0216] The pharmaceutical compositions of the present invention can be formulated for parenteral administration, for example, for injection via intravenous, intramuscular, subcutaneous, intra-wound, or intraperitoneal routes. According to this disclosure, the preparation of aqueous compositions, such as aqueous pharmaceutical compositions containing the compounds described herein, is known to those skilled in the art. Typically, such compositions can be prepared as injections, as liquid solutions or suspensions; they can also be prepared in a solid form suitable for addition to a liquid prior to injection to prepare a solution or suspension; and the preparations can also be emulsified.

[0217] Suitable forms of drugs for injection include sterile aqueous solutions or dispersants; formulations containing sesame oil, peanut oil, or aqueous propylene glycol; and sterile powders for the ad hoc preparation of sterile injectable solutions or dispersants. In all cases, the form must be sterile and must be a fluid to facilitate injection. It must be stable under manufacturing and storage conditions and must be protected against contamination by microorganisms such as bacteria and fungi.

[0218] Solutions of active compounds or pharmaceutically acceptable salts as free bases can be prepared in water by suitably mixing with surfactants such as hydroxypropyl cellulose. Dispersants can also be prepared in glycerol, liquid polyethylene glycol and mixtures thereof, and in oils. Furthermore, sterile, non-volatile oils can be used as solvents or suspension media. For this purpose, any mild, non-volatile oil can be used, including synthetic monoglycerides or diglycerides of glycerol. Additionally, fatty acids such as oleic acid can be used to prepare injectable preparations. Sterile injectable preparations can also be sterile injectable solutions, suspensions, or emulsions present in non-toxic, parenteral-acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable carriers and solvents that can be used are water, Ringer's solution, USP, and isotonic sodium chloride solution. In one embodiment, the compound can be suspended in a solution containing 1% (w / v) sodium carboxymethyl cellulose and 0.1% (v / v) TWEEN. TM In a carrier fluid of 80%. Under normal storage and use conditions, these preparations contain preservatives to inhibit microbial growth.

[0219] Injectable formulations, such as sterile injectable aqueous or oily suspensions, can be formulated using suitable dispersants or wetting agents and suspending agents according to known techniques. Typically, dispersants are prepared by incorporating various sterile active ingredients into a sterile carrier containing a basic dispersion medium and other desired components from those listed above. The sterile injectable solutions of the present invention can be prepared by incorporating the compounds described herein with appropriate amounts of the aforementioned other components as needed, followed by filtration and sterilization. In the case of preparing sterile powders for sterile injectable solutions, preferred methods of preparation include vacuum drying and freeze-drying techniques, which generate powders of the active ingredients plus any additional desired components from their previously sterilized and filtered solutions. Injectable formulations can be sterilized, for example, by filtration through a bacterial retention filter.

[0220] It is also considered to prepare larger or highly concentrated solutions for intramuscular injection. In this regard, the use of DMSO as a solvent is preferred because it will result in very rapid penetration, delivering high concentrations of the compound to a small area.

[0221] Suitable preservatives for such solutions include benzalkonium chloride, benzyl chloride, chlorobutanol, thimerosal, etc. Suitable buffers include boric acid, sodium bicarbonate and potassium bicarbonate, sodium borate and potassium borate, sodium carbonate and potassium carbonate, sodium acetate, disodium hydrogen phosphate, etc., in amounts sufficient to maintain the pH between approximately pH 6 and pH 8, preferably between approximately pH 7 and pH 7.5. Suitable osmolarity modifiers are dextran 40, dextran 70, glucose, glycerol, potassium chloride, propylene glycol, sodium chloride, etc., such that the sodium chloride equivalent of the solution is in the range of 0.9 ± 0.2%. Suitable antioxidants and stabilizers include sodium bisulfite, sodium metabisulfite, sodium thiosulfite, thiourea, etc. Suitable wetting and clarifying agents include polysorbate 80, polysorbate 20, poloxamer 282, and tyloxapine. Suitable thickeners include dextran 40, dextran 70, gelatin, glycerin, hydroxyethyl cellulose, hydroxymethylpropyl cellulose, lanolin, methylcellulose, petrolatum, polyethylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, carboxymethyl cellulose, etc.

[0222] In some embodiments, this document includes compositions suitable for oral delivery of the compounds described herein, such as tablets comprising an enteric coating, such as an acid-resistant coating, such that the composition can deliver the compounds to, for example, the gastrointestinal tract of a patient.

[0223] For example, tablets for oral administration are provided, comprising particles (e.g., at least partially formed of particles) and one or more pharmaceutically acceptable excipients, said particles including the compounds described herein. Such tablets may be coated with enteric coating. The contemplated tablets may include pharmaceutically acceptable excipients such as fillers, binders, disintegrants and / or lubricants, as well as colorants, release agents, coating agents, sweeteners, flavoring agents (such as wintergreen, orange, xylitol, sorbitol, fructose, and maltodextrin), and flavoring agents, preservatives, and / or antioxidants.

[0224] In some embodiments, the contemplated pharmaceutical formulation includes an internal phase and a pharmaceutically acceptable filler, said internal phase comprising the compounds described herein or pharmaceutically acceptable salts thereof. For example, the compounds and fillers may optionally be mixed with other excipients to form granules. In some embodiments, the internal phase may be formed using a wet granulation process, such as adding a liquid (e.g., water) to a blend of compounds and fillers, followed by drying, grinding, and / or sieving the composition to produce granules. Those skilled in the art will understand that other methods can be used to achieve the internal phase.

[0225] In some embodiments, the intended formulation includes an exophase that may include one or more pharmaceutically acceptable excipients and may be blended with the inophase to form the disclosed formulation.

[0226] The disclosed formulation may include an intraparticle phase containing a filler. Exemplary fillers include, but are not limited to, cellulose, gelatin, calcium phosphate, lactose, sucrose, glucose, mannitol, sorbitol, microcrystalline cellulose, pectin, polyacrylate, glucose, cellulose acetate, hydroxypropyl methylcellulose, partially pregelatinized starch, calcium carbonate, and others, including combinations thereof.

[0227] In some embodiments, the disclosed formulation may include an internal and / or external phase containing a binder, which is typically used to hold the components of a pharmaceutical formulation together. Exemplary binders of the present invention may include, but are not limited to, substances such as starch, sugar, cellulose or modified cellulose such as hydroxypropyl cellulose, lactose, pregelatinized corn starch, polyvinylpyrrolidone, hydroxypropyl cellulose, hydroxypropyl methylcellulose, low-substituted hydroxypropyl cellulose, sodium carboxymethyl cellulose, methylcellulose, ethyl cellulose, sugar alcohols, and others, including combinations thereof.

[0228] The intended formulation, such as a formulation comprising an internal phase and / or an external phase, may include a disintegrant, such as, but not limited to, starch, cellulose, cross-linked polyvinylpyrrolidone, sodium starch glycolate, sodium carboxymethyl cellulose, alginate, corn starch, cross-linked sodium carboxymethyl cellulose, cross-linked carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, gum arabic, and others, including combinations thereof. For example, the internal phase and / or external phase may include a disintegrant.

[0229] In some embodiments, the intended formulation comprises an internal phase and an external phase, the internal phase comprising the compounds and excipients described herein, the excipients being selected from mannitol, microcrystalline cellulose, hydroxypropyl methylcellulose, and sodium glycolate, or combinations thereof, and the external phase comprising one or more of microcrystalline cellulose, sodium glycolate starch, and magnesium stearate, or mixtures thereof.

[0230] In some embodiments, the contemplated formulation may include a lubricant, for example, the particulate phase may contain a lubricant. Lubricants include, but are not limited to, talc, silica, fats, stearin, magnesium stearate, calcium phosphate, silica, calcium silicate, calcium phosphate, colloidal silica, metal stearates, hydrogenated vegetable oils, corn starch, sodium benzoate, polyethylene glycol, sodium acetate, calcium stearate, sodium lauryl sulfate, sodium chloride, magnesium lauryl sulfate, talc, and stearic acid.

[0231] In some embodiments, the pharmaceutical formulation comprises an enteric coating. Typically, an enteric coating creates a barrier for orally administered drugs, controlling where the drug is absorbed along the digestive tract. Enteric coatings can include polymers that decompose at different rates depending on pH. Examples of enteric coatings include, for instance, cellulose acetate phthalate, methyl acrylate-methacrylic acid copolymer, cellulose acetate succinate, hydroxypropyl methylcellulose phthalate, methyl methacrylate-methacrylic acid copolymer, ethyl acrylate-methacrylic acid copolymer, C-type methacrylic acid copolymer, polyvinyl acetate-phthalate, and cellulose acetate phthalate.

[0232] Exemplary enteric coatings include AMB, Acryl- These levels. In some embodiments, the enteric coating may comprise about 5% to about 10%, about 5% to about 20%, 8% to about 15%, about 8% to about 20%, about 10% to about 20%, or about 12% to about 20% or about 18% of the expected tablet weight. For example, the enteric coating may comprise an ethyl acrylate-methacrylic acid copolymer.

[0233] For example, in one anticipated embodiment, a tablet is provided comprising or substantially composed of: about 0.5% to about 70%, for example about 0.5% to about 10%, or about 1% to about 20% by weight of the compound described herein or a pharmaceutically acceptable salt thereof. Such a tablet may comprise, for example, about 0.5% to about 60% by weight of mannitol, for example about 30% to about 50% by weight of mannitol, for example about 40% by weight of mannitol; and / or about 20% to about 40% by weight of microcrystalline cellulose, or about 10% to about 30% by weight of microcrystalline cellulose. For example, the disclosed tablets may include an intraparticle phase comprising about 30% to about 60% by weight, for example about 45% to about 65% by weight, or about 5% to about 10% by weight of the compounds described herein, about 30% to about 50% by weight, or alternatively about 5% to about 15% by weight of mannitol, about 5% to about 15% by weight of microcrystalline cellulose, about 0% to about 4% or about 1% to about 7% by weight of hydroxypropyl methylcellulose, and about 0% to about 4% by weight, for example about 2% to about 4% by weight of sodium glycolate starch.

[0234] In another anticipated embodiment, a pharmaceutical tablet formulation for oral administration of the compound described herein comprises an internal phase comprising the compound described herein or a pharmaceutically acceptable salt (e.g., a sodium salt) and a pharmaceutically acceptable filler, and may further comprise an external phase comprising pharmaceutically acceptable excipients, such as disintegrants. The external phase may comprise components selected from microcrystalline cellulose, magnesium stearate, and mixtures thereof. The pharmaceutical composition may also comprise an enteric coating of about 12% to 20% by weight of the tablet. For example, a pharmaceutically acceptable tablet for oral use may comprise about 0.5% to 10% by weight of the compound described herein or a pharmaceutically acceptable salt thereof, about 30% to 50% by weight of mannitol, about 10% to 30% by weight of microcrystalline cellulose, and an enteric coating comprising ethyl acrylate-methacrylic acid copolymer.

[0235] In another example, a pharmaceutically acceptable tablet for oral use may comprise an internal phase comprising about 5 to about 10% by weight of the compound described herein or a pharmaceutically acceptable salt thereof, about 40% by weight of mannitol, about 8% by weight of microcrystalline cellulose, about 5% by weight of hydroxypropyl methylcellulose, and about 2% by weight of sodium starch glycolate; an external phase comprising about 17% by weight of microcrystalline cellulose, about 2% by weight of sodium starch glycolate, and about 0.4% by weight of magnesium stearate; and an enteric coating comprising ethyl acrylate-methacrylic acid copolymer on the tablet.

[0236] In some embodiments, the pharmaceutical composition may contain an enteric coating comprising about 13% or about 15%, 16%, 17% or 18% by weight, for example... (See, for example, PCT Publication No. WO2010 / 054826, which is incorporated herein by reference in its entirety.)

[0237] The rate at which the coating dissolves and the active ingredient is released at a given point is its dissolution rate. In one embodiment, the intended tablet may have a dissolution characteristic, for example, when tested in a Type 2 USP / EP device (paddle) at 100 rpm and 37°C in phosphate buffer at pH 7.2, about 50% to about 100% of the compound described herein is released after about 120 minutes to about 240 minutes, e.g., 180 minutes. In another embodiment, the intended tablet may have a dissolution characteristic, for example, when tested in a Type 2 USP / EP device (paddle) at 100 rpm and 37°C in diluted HCl at pH 1.0, where substantially no compound is released after 120 minutes. In yet another embodiment, the intended tablet may have a dissolution characteristic, for example, when tested in a Type 2 USP / EP device (paddle) at 100 rpm and 37°C in phosphate buffer at pH 6.6, about 10% to about 30%, or no more than about 50%, of the compound is released after 30 minutes.

[0238] In some embodiments, the methods provided herein may further include administering at least one other agent for the treatment of the conditions, diseases, and illnesses disclosed herein. In one embodiment, the contemplated other agents may be administered (e.g., sequentially or simultaneously) together.

[0239] Non-limiting examples of such combination therapies include combining one or more of the compounds of the present invention with anti-inflammatory agents, antibiotics, immunosuppressants, immunomodulators, or analgesics.

[0240] The anticipated agents include immunosuppressants, such as glucocorticoids, cytosolic inhibitors, antibodies, agents acting on immunosuppressants, interferons, opioids, TNF-binding proteins, mycophenolate mofetil, and small biological agents. For example, anticipated immunosuppressants include, but are not limited to: tacrolimus, cyclosporine, pimecrolimus, sirolimus, everolimus, mycophenolate mofetil, fingolimod, dexamethasone, fludarabine, cyclophosphamide, methotrexate, azathioprine, leflunomide, teriflunomide, anaraxine, anti-thymocyte globulin, anti-lymphocyte globulin, moromuzumab-CD3, afutocilizumab, rituximab, tibolizumab, efazolin, dalizumab, baliximab, adalimumab, infliximab, sertocilizumab-butazol, natezil, and etanercept. Other anticipated medications include antidiarrheal drugs, laxatives, iron supplements, and calcium or vitamin D or B-12 supplements.

[0241] Exemplary formulations include dosage forms that comprise or consist substantially of the compound described herein, from about 35 mg to about 500 mg. For example, formulations may comprise about 35 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, or 250 mg of the compound described herein. In one embodiment, the formulation may comprise about 40 mg, 80 mg, or 160 mg of the compound described herein. In some embodiments, the formulation may comprise at least 100 μg of the compound described herein. For example, the formulation may comprise about 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, or 25 mg of the compound described herein. The dosage will depend on variables such as the type and severity of the disease or indication to be treated, the patient's overall health and body size, the compound's efficacy in vivo, the drug formulation, and the route of administration. The initial dose may be increased beyond the upper limit to rapidly reach the desired blood or tissue levels. Alternatively, the initial dose may be less than the optimal dose, and the dose may be gradually increased during treatment. Dosing frequency can vary depending on factors such as the route of administration, dosage, and the disease being treated. Exemplary dosing frequencies are once daily, once weekly, and once every two weeks. In some embodiments, dosing is once daily for seven consecutive days. Example

[0242] The present invention is further illustrated by the following embodiments. These embodiments are provided for illustrative purposes only and should not be construed as limiting the scope or content of the invention in any way.

[0243] Example 1: Synthesis of Sample 2

[0244] Sample 2 was prepared according to Scheme 1 as shown below. The compound numbers used in this embodiment are only applicable to this embodiment; sample 2 is referred to as compound 2 elsewhere.

[0245] Option 1

[0246]

[0247] Synthesis of 2-(2-(triethylsilyl)-5-(trifluoromethyl)-1H-indol-3-yl)ethanol (3)

[0248] A mixture of 50 mL of N,N-dimethylformamide (DMF) containing 2-iodo-4-(trifluoromethyl)aniline (1) (5.00 g, 17.4 mmol), 4-(triethylsilyl)but-3-yn-1-ol (2) (3.85 g, 1.2 mmol), [bis(diphenylphosphino)ferrocene]palladium(II) chloride (0.64 g, 0.87 mmol), lithium chloride (0.732 g, 17.4 mmol), and sodium carbonate (3.7 g, 34.8 mmol) was stirred at 100 °C for 15 hours. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic extract was dried over Na₂SO₄ and concentrated under reduced pressure. The crude material was purified by silica gel chromatography to give 7.00 g of the title compound as a yellow oil containing ~20% of the starting material (2). The product was used in subsequent steps without further purification. 1 HNMR (300MHz, CDCl3) δ8.2 (bs, 1H), 7.9 (s, 1H), 7.45 (s, 2H), 3.85 (q, J = 8.0Hz, 2H), 3.15(t,J=6.4Hz,2H), 1.46(t,J=6.0Hz,1H), 1.2(m,15H); MS(APCI+)m / z=344(M+H).

[0249] Synthesis of 2-(5-(trifluoromethyl)-1H-indol-3-yl)ethanol (4)

[0250] A solution of 15 mL of 2-(2-(triethylsilyl)-5-(trifluoromethyl)-1H-indol-3-yl)ethanol (3) (2.00 g, 5.83 mmol) in tetrahydrofuran (THF) was added to tetrabutylammonium fluoride (7.0 mL, 1 M, soluble in THF), and the reaction mixture was stirred at room temperature (rt) for 72 hours. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic extract was dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by silica gel chromatography to give 1.02 g (75%) of the title compound as a pale yellow oil. 1 HNMR (300MHz, CDCl3) δ8.3 (bs, 1H), 7.96 (s, 1H), 7.45 (s, 2H), 7.2 (s, 1H), 3.90 (t, J = 6.8Hz, 2H), 3.05 (t, J = 6.8Hz, 2H); MS (APCI+) m / z = 230 (M + H).

[0251] Synthesis of 3-(2-bromoethyl)-5-(trifluoromethyl)-1H-indole (5)

[0252] A THF solution (10 mL) of 2-(5-(trifluoromethyl)-1H-indol-3-yl)ethanol (4) (1.00 g, 4.36 mmol) was added to a THF solution (10 mL) of triphenylphosphine (2.30 g, 8.72 mmol) and perbromomethane (4.40 g, 13.1 mmol) pre-stirred for 1 hour. The resulting mixture was stirred at room temperature for 3 hours. The reaction mixture was then filtered and concentrated under reduced pressure. The crude material was purified by silica gel chromatography to give 0.66 g (51%) of the title compound as a yellow oil. 1 HNMR (300MHz, CDCl3) δ8.25 (bs, 1H), 7.9 (s, 1H), 7.4 (s, 2H), 7.2 (s, 1H), 3.7 (t, J = 7.8Hz, 2H), 3.27 (t, J = 7.8Hz, 2H).

[0253] Synthesis of 3-(2-azidoethyl)-5-(trifluoromethyl)-1H-indole (6)

[0254] A mixture of 3-(2-bromoethyl)-5-(trifluoromethyl)-1H-indole (5) (0.66 g, 2.26 mmol) and sodium azide (0.44 g; 6.8 mmol) in DMF (10 mL) was stirred at 70 °C for 4 hours. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic layer was washed successively with brine and sodium thiosulfate, dried, and concentrated under reduced pressure. The crude substance was purified by silica gel chromatography to give 0.61 g (100%) of the title compound as a brown oil. 1 HNMR (300MHz, CDCl3) δ8.15 (bs, 1H), 7.87 (s, 1H), 7.44 (d, J = 1.5Hz, 2H), 7.19 (s, 1H), 3.55-3.61 (t, J = 7Hz, 2H), 3.08 (t, J = 7.2Hz, 2H).

[0255] Synthesis of 2-(5-(trifluoromethyl)-(1H-indol-3-yl))ethylamine (7)

[0256] A mixture of 3-(2-azidoethyl)-5-(trifluoromethyl)-H-indole (6) (0.61 g, 2.45 mmol) and triphenylphosphine (1.93 g, 7.41 mmol) in methanol (10 mL) was stirred at 70 °C for 2 hours. The reaction mixture was concentrated under reduced pressure, and the crude substance was purified by silica gel chromatography to give 0.44 g (75%) of the title compound as a brown oil. 1HNMR (300MHz, CDCl3) δ8.4 (bs, 1H), 7.9 (s, 1H), 7.4 (s, 2H), 7.25 (s, 1H), 7.13-7.14 (d, J = 2.4Hz, 1H), 3.04 (m, 4H); MS (APCI-) m / z = 326 (MH).

[0257] Synthesis of 1-(3,4-dimethoxyphenyl)-6-(trifluoromethyl)-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]in Dole (sample 2)

[0258] A mixture of 2-(5-(trifluoromethyl)-1H-indol-3-yl))ethylamine (7) (0.40 g, 1.76 mmol) and 3,4-dimethoxybenzaldehyde (8) (0.322 g, 1.93 mmol) in acetic acid solution (8 ml) was stirred at 80 °C for 24 hours. The reaction mixture was concentrated under reduced pressure, and the crude product was purified by silica gel chromatography to give 0.16 g (33%) of the title compound as a white solid. 1 HNMR (300MHz, CDCl3) δ7.9 (s, 1H), 7.71 (bs, 1H), 7.33 (m, 1H), 6.84 (d, J = 3.8Hz, 2H), 5.1 2(s,1H),3.88(s,3H),3.81(s,3H),3.15(m,1H),2.88(m,2H); MS(APCI+)m / z=377(M+H).

[0259] Example 2: Synthesis of Sample 4a

[0260] Sample 4a was prepared according to Scheme 2 as shown below. The compound numbers used in this embodiment are only applicable to this embodiment; sample 4a is referred to as compound 4a elsewhere.

[0261] Option 2

[0262]

[0263] Synthesis of 2-iodo-1-methyl-1H-indole (2)

[0264] At 0 °C, a THF solution (20 mL) of 2-iodo-1H-indole (1) (1.50 g, 6.17 mmol) was added to a suspension of 60% NaH (0.37 g, 9.25 mmol), and the resulting solution was stirred for 10 minutes. Methyl iodine (1.75 g, 12.3 mmol) was added dropwise, and the reaction mixture was slowly heated from 0 °C to room temperature over 1 hour. The reaction was quenched with saturated NH4Cl solution (15 mL) and extracted with ethyl acetate. The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by silica gel chromatography to give 1.27 g (80%) of the title compound as a pale yellow oil.1 HNMR (400MHz, CDCl3) δ7.71(d,J=7.6Hz,1H),7.29(d,J=8Hz,1H),7.14(t,J=7.6Hz,1H),7.06(t,J=7.6Hz,1H),6.78(s,1H),3.74(s,3H).

[0265] Synthesis of 3-(2-iodo-1-methyl-1H-indol-3-yl)propionaldehyde (4)

[0266] TFA (0.16 g, 1.48 mmol) was added dropwise to a stirred solution of acrolein (3) (1.38 g, 24.7 mmol) and N-methylaniline (0.16 g, 1.48 mmol) at 0 °C, and the reaction mixture was stirred at 0 °C for 10 min. 2-Iodo-1-methyl-1H-indole (2) (1.27 g, 4.94 mmol) was added to CH₂Cl₂ (4 mL), and the reaction mixture was stirred slowly for 3 h, then brought to room temperature from 0 °C. The reaction mixture was diluted with water and extracted with dichloromethane (DCM). The organic layer was dried over Na₂SO₄ and concentrated under reduced pressure. The crude material was purified by silica gel chromatography to give 920 mg (59%) of the title compound as a pale yellow foam. 1 HNMR (400MHz, CDCl3) δ9.85(s,2H),7.51(d,J=8.0Hz,1H),7.30(d,J=8.4Hz,1H),7.16(t,J=9.8Hz,1H),7.08( t,J=8Hz,1H),3.75(s,3H),3.75(s,3H),3.09(t,J=7.6Hz,2H),2.76(t,J=8.2Hz,2H); MS(ESI+)m / z=314(M+H).

[0267] Synthesis of N-(3-(2-iodo-1-methyl-1H-indol-3-yl))propyl-3,4,5-trimethoxyaniline (6)

[0268] One drop of acetic acid was added to a methanol solution (15 mL) of 3-(2-iodo-1-methyl-1H-indol-3-yl)propionaldehyde (4) (920 mg, 2.93 mmol) and 3,4,5-trimethoxyaniline (5) (806 mg, 4.40 mmol), and the reaction mixture was stirred for 10 minutes. NaCNBH3 (0.46 g, 7.32 mmol) was added, and stirring was continued for 16 hours. The reaction mixture was diluted with water and extracted with DCM. The organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by silica gel chromatography to give 1.07 g (74%) of the title compound as a grayish-white foam. 1HNMR (400MHz, CDCl3) δ7.53(d,J=7.8Hz,1H),7.31(d,J=8.1Hz,1H),7.16(t,J=9Hz,1H),7.06(t,J=7.8Hz,1H), 5.77 (s, 2H), 3.75 (s, 3H), 3.14 (t, J = 7.05Hz, 2H), 2.88 (t, J = 7.2Hz, 2H), 1.26 (m, 2H); MS (ESI+) m / z = 481 (M + H).

[0269] Synthesis of 9-methyl-1-(3,4,5-trimethoxyphenyl)-2,3,4,9-tetrahydro-1H-pyrido[2,3-b]indole (Sample 4a)

[0270] To a toluene solution (5 mL) of N-(3-(2-iodo-1-methyl-1H-indol-3-yl))propyl)-3,4,5-trimethoxyaniline (6) (1.07 g, 2.22 mmol), tris(dibenzylacetone)dipalladium (0) (0.1 g, 0.108 mmol), 2,2'-bis(diphenylphosphino)-1,1”-binaphthyl (0.12 g, 0.216 mmol), and t-BuONa (0.21 g, 2.16 mmol) were added, and the reaction mixture was stirred at 100 °C for 2 hours. The mixture was concentrated under reduced pressure, and the crude product was purified by silica gel chromatography to give 280 mg (36%) of the title compound as a white solid. 1 HNMR(400MHz, CDCl3)δ7.45(d,1H),7.15(m,3H),6.19(s,2H),3.82(s,3H),3.71( m, 8H), 3.26 (s, 3H), 2.79 (t, J = 6.6Hz, 2H), 1.87 (m, 2H); MS (ESI+) m / z = 353 (M+H).

[0271] Example 3: Synthesis of Sample 13

[0272] Sample 13 was prepared according to Scheme 3 as shown below. The compound numbers used in this embodiment are only applicable to this embodiment; sample 13 is referred to as compound 13 elsewhere.

[0273] Option 3

[0274]

[0275] Synthesis of 3-bromothiophene-2-carboxaldehyde (3)

[0276] 3-Bromothiophene (3.0 g, 31 mmol) was added to 50 mL of a THF solution of lithium diisopropylamino (31.6 mmol) at -78 °C (1). After stirring for 1 hour, formylpiperidine (3.50 g, 31.6 mmol) was added (2), and the reaction was heated to 0 °C. After 12 hours, the reaction mixture was partitioned between DCM and saturated NH4Cl, and the organic layer was separated, dried over MgSO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography to give 1.90 g (54%) of the title compound as a yellow oil. 1 HNMR (400MHz, CDCl3) δ9.99 (s, 1H), 7.71 (d, J = 5.2Hz, 1H), 7.15 (d, J = 4.8Hz, 1H).

[0277] Synthesis of 3-azidothiophene-2-carboxaldehyde (4)

[0278] A solution of 3-bromothiophene-2-carboxaldehyde (3) (1.90 g, 0.99 mmol) and sodium azide (3.23 g, 50 mmol) in 50 mL of 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU) was heated to 50 °C and stirred for 36 hours. The reaction mixture was poured over ice water and then extracted with DCM. The organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography to give 1.00 g (66%) of the title compound as a yellow oil. 1 HNMR (400MHz, CDCl3) δ9.93 (s, 1H), 7.72 (d, J = 5.2 Hz, 1H), 7.04 (d, J = 5.2 Hz, 1H).

[0279] Synthesis of (E)-3-(3-azidothiophen-2-yl)-1-(3,4,5-trimethoxyphenyl)prop-2-en-1-one (6)

[0280] To a mixture of 3-azidothiophene-2-carboxaldehyde (4) (0.80 g, 5.22 mmol) and 1-(3,4,5-trimethoxyphenyl)ethyl ketone (5) (1.64 g, 7.84 mmol) in MeOH (5 mL), 2 mL of an aqueous solution of NaOH (0.62 g, 15.66 mmol) was added, and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with water and extracted with DCM. The organic extract was dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude material was purified by silica gel chromatography to give 1.10 g (61%) of the title compound as a yellow solid. 1HNMR(400MHz, CDCl3) δ7.88(d,J=15.6Hz,1H),7.42(d,J=5.2Hz,1H),7.21(d,J=1 5.2Hz, 3H), 7.00 (d, J = 5.2Hz, 1H), 3.94 (d, J = 5.2Hz, 9H); MS (ESI+) m / z = 346 (M+H).

[0281] Synthesis of (4H-thieno[3,2-b]pyrrolo-5-yl)(3,4,5-trimethoxyphenyl) methyl ketone (7)

[0282] A 10 mL xylene solution of (E)-3-(3-azidothiophen-2-yl)-1-(3,4,5-trimethoxyphenyl)prop-2-en-1-one (6) (1.20 g, 3.48 mmol) was stirred at 150 °C for 30 min. The reaction mixture was concentrated under reduced pressure, and the crude product was purified by silica gel chromatography to give 1.00 g (91%) of the title compound as a grayish-white solid. 1 HNMR (300MHz, CDCl3) δ9.48 (bs, 1H), 7.42 (d, J = 5.4Hz, 1H), 7.20 (s, 2H), 7.11 (s, 1H), 7.01 (d, J = 5.7Hz, 1H), 3.94 (d, J = 1.8Hz, 9H); MS (ESI+) m / z = 318 (M + H).

[0283] Synthesis of 5-(3,4,5-trimethoxybenzyl)-4H-thieno[3,2-b]pyrrole (Sample 13)

[0284] At 100 °C, a mixture of 2-propanol (10 mL) of (4H-thieno[3,2-b]pyrrolo-5-yl)(3,4,5-trimethoxyphenyl) methyl ketone (7) (0.80 g; 2.52 mmol) and sodium borohydride (480 mg, 12.7 mmol) was stirred in a sealed tube for 2 hours. The reaction mixture was cooled, diluted with water, and extracted with DCM. The organic extract was dried over Na2SO4 and concentrated. The crude material was purified by silica gel chromatography to give 280 mg (37%) of the title compound as a white solid. 1 HNMR (300MHz, CDCl3) δ7.92 (bs, 1H), 6.99 (d, J = 5.1Hz, 1H), 6.86 (d, J = 5.1Hz, 1H), 6. 46 (s, 2H), 6.26 (s, 1H), 4.02 (s, 2H), 3.83 (t, J = 3.75Hz, 9H); MS (ESI+) m / z = 304 (M + H).

[0285] Example 4: Synthesis of Sample 15

[0286] Sample 15 was prepared according to Scheme 4 as shown below. The compound numbers used in this embodiment are only applicable to this embodiment; sample 15 is referred to as compound 15 elsewhere.

[0287] Option 4

[0288]

[0289] Synthesis of 6-(trifluoromethyl)benzo[d]thiazole (3)

[0290] Amyl nitrite (3.22 g, 27.5 mmol) was added to 20 mL of a THF solution of 6-(trifluoromethyl)benzothiazol-2-amine (1) (2.00 g, 9.17 mmol). The mixture was heated to reflux for 30 min, quenched with water, and extracted with ethyl acetate. The organic extract was dried over Na2SO4 and concentrated under reduced pressure. The crude substance was purified by silica gel chromatography to give 860 mg (46%) of the title compound as a yellow solid. 1 HNMR (400MHz, CDCl3) δ9.16 (s, 1H), 8.26 (m, 2H), 7.78 (d, J = 1.2Hz, 1H).

[0291] Synthesis of 2-amino-5-(trifluoromethyl)thiophenol (4)

[0292] A solution of 6-(trifluoromethyl)benzo[d]thiazole (3) (830 mg, 4.08 mmol) and hydrazine monohydrate (1.52 g, 30.6 mmol) in ethanol (20 mL) was heated to reflux for 1.5 hours. The mixture was added to an aqueous solution of acetic acid (3 mL) (100 mL) and extracted with DCM. The organic extract was dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by silica gel chromatography to give 670 mg (84%) of the title compound as a yellow oil. 1 HNMR (400MHz, CDCl3) δ7.62 (d, J = 1.2 Hz, 1H), 7.39 (dd, J = 8.4, 2.0 Hz, 1H), 6.75 (d, J = 8.4 Hz, 1H), 4.49 (s, 2H), 2.95 (s, 1H).

[0293] Synthesis of 2-(3,4-dimethoxybenzyl)-6-(trifluoromethyl)benzo[d]thiazole (Sample 15)

[0294] 2-Amino-5-(trifluoromethyl)benzylthiol (4) (400 mg, 2.07 mmol), 2-(3,4-dimethoxyphenyl)acetic acid (5) and Lawson's reagent (0.29 g, 0.72 mmol) were placed in a sealed tube and microwaved at 190 °C for 5 minutes. The crude substance was purified by silica gel chromatography to give 320 mg (43%) of the title compound as a white solid. 1HNMR (400MHz, CDCl3) δ8.08 (s, 1H), 8.06 (s, 1H), 7.69 (d, J = 7.5Hz, 1H), 6.91 (m, 3H), 4.40 (s, 1H), 3.87 (d, J = 6.0Hz, 6H); MS (ESI+) m / z = 354 (M + H).

[0295] Example 5: Synthesis of Sample 17

[0296] Sample 17 was prepared according to Scheme 5 as shown below. The compound numbers used in this embodiment are only applicable to this embodiment; sample 17 is referred to as compound 17 elsewhere.

[0297] Option 5

[0298]

[0299] Synthesis of N-(3,4,5-trimethoxyphenyl)-1H-benzo[d]imidazol-2-amine (Sample 17)

[0300] A mixture of 2-chloro-1H-benzis[d]imidazole (1) (0.40 g, 2.63 mmol), potassium dihydrogen phosphate (0.36 g, 2.63 mmol), and 3,4,5-trimethoxyaniline (0.48 g, 2.63 mmol) in 20 mL of n-BuOH was stirred at 90 °C for 16 hours. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic extract was dried over Na₂SO₄ and concentrated under reduced pressure. The crude product was purified by silica gel chromatography to give 0.43 g (54%) of the title compound as a grayish-white solid. 1 HNMR (300MHz, CDCl3) δ10.8(s,1H),9.29(s,1H),7.28(bs,2H),7.12(s,2H),6.97(bs,2H),3.79(s,6H),3.62(s,3H); MS(ESI+)m / z=300(M+H).

[0301] Example 6: Synthesis of Sample 24

[0302] Sample 24 was prepared according to Scheme 6 as shown below. The compound numbers used in this embodiment are only applicable to this embodiment; sample 24 is referred to as compound 24 elsewhere.

[0303] Option 6

[0304]

[0305] Synthesis of thiophene-2-amine·TFA salt (2)

[0306] 2,2,2-trifluoroacetic acid (1.43 g, 12.55 mmol) was added to 10 mL of a DCM solution of tert-butylthiophene-2-ylcarbamate (1) (0.50 g, 2.51 mmol). The mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure to give ~0.50 g of the title compound, which was used directly in the next step without purification.

[0307] Synthesis of 2-(3,4,5-trimethoxyphenyl)acetyl chloride (4)

[0308] Oxaloyl chloride (0.67 g, 5.30 mmol) was added to an ice-cold solution of 2-(3,4,5-trimethoxyphenyl)acetic acid (3) (0.40 g, 1.77 mmol) in 10 mL of DCM, followed by 1 drop of DMF. The mixture was stirred for 2 hours from 0 °C to room temperature. The reaction mixture was concentrated under reduced pressure to give ~0.50 g of the title compound, which was used in the next step without purification.

[0309] Synthesis of N-(thiophen-2-yl)-2-(3,4,5-trimethoxyphenyl)acetamide (Sample 24)

[0310] To an ice-cold solution of 10 mL of DCM containing thiophene-2-amine·TFA salt (2) (~0.50 g, crude) and diisopropylethylamine (DIPEA) (0.99 g, 7.65 mmol), 5 mL of DCM solution of 2-(3,4,5-trimethoxyphenyl)acetyl chloride (4) (~0.50 g, crude) was added, and the reaction mixture was stirred for 2 hours from 0 °C to room temperature. The reaction mixture was diluted with water and extracted with DCM. The organic extract was dried over Na2SO4 and concentrated under reduced pressure. The crude substance was purified by silica gel chromatography to give 0.25 g of the title compound as a white solid. 1 HNMR (300MHz, CDCl3) δ7.80 (bs, 1H), 6.87 (m, 1H), 6.81 (m, 1H), 6.56 (m, 1H), 6.51 (s, 2H), 3.86 (s, 9H), 3.70 (s, 2H); MS (ESI+) m / z = 308 (M+H).

[0311] Example 7: Design and in vitro activity of a selective Ah receptor modulator (SAhRM) exhibiting anti-inflammatory properties

[0312] As a starting point for developing the compounds disclosed in this invention, the compounds belong to a series of 1-aryl-1,2,3,4-tetrahydro-β-carbaline derivatives ( Figure 1 A group of 47 compounds were evaluated for their ability to increase IL-22 production and decrease IFN-γ production in vitro. Figure 23 ). Figure 24In vitro results for some of the above-mentioned compounds are shown. These compounds reduce the production of IFN-γ. Notably, in addition to reducing IFN-γ production, compounds 15 and 17, for example, also strongly increase the production of IL-22.

[0313] Conformational analysis

[0314] Conformational analysis was performed on all compounds in these series to determine the spatial positions of their substituents and to identify their putative bioactive conformations among the lowest-energy conformational isomers.

[0315] Molecular modeling was performed using SYBYL software version 6.92. Three-dimensional models of all compounds were built from the standard fragment library, and their geometries were subsequently optimized using a Tripos force field incorporating electrostatic terms calculated from Gasteiger and Hückel atomic charges. The Powell method, available in the Maximin2 program, was used for energy minimization until the gradient value was less than 0.0001 kcal / mol.

[0316] For each compound, a conformational search was performed using a randomized search process implemented in SYBYL to identify its lowest-energy conformation. Randomized conformational search is a technique for locating the minimum energy value of a molecule. It involves randomly twisting selected chemical bonds, followed by minimization. This cycle of random twisting and minimization is repeated multiple times. After each cycle, the new conformation is compared with other conformations discovered so far to see if it is unique. The primary option used for the randomized search is maximum hit (n=6), which defines the minimum number of times each conformation must be found to stop the search for new conformations, and an RMS threshold. It is defined as the maximum RMS difference between two conformations before they are considered different.

[0317] The conformations generated by the random conformation search were fully optimized, and their geometries could be immediately re-optimized using these conformations. A semi-empirical MOPAC package of version 6.0 with Hamiltonian AM1 (keywords: PRECISE, NOMM, PARASOK) was used, and the Coulson partial atomic charges were calculated using the same method. Table 1 shows the conformational space of the selected compounds.

[0318] Table 1

[0319] compound n <![CDATA[E min (kcal)]]> <![CDATA[E max (kcal)]]> CTL-01-07-L-A03 2 80.76 83.82 CTL-01-07-L-A04 16 43.15 47.70 CTL-01-07-L-B05 157 26.34 61.71 CTL-10-26-L-C08 126 -5.12 30.49 CTL-10-26-L-C09 66 -27.40 -10.60 CTL-10-26-L-C10 26 31.33 40.74 CTL-10-26-L-F07 30 29.75 47.35 CTL-10-26-L-F09 97 26.64 51.86 CTL-10-26-L-G10 135 -40.40 -3.65 CTL-10-26-L-H04 202 -8.80 20.56 CTL-10-26-L-H05 106 26.05 45.80 CTL-10-26-L-H06 49 8.01 13.62 CTL-10-29-L-A05 253 -43.79 -7.10 SGA360 853 -95.57 -63.71 Leflunomide 29 -124.59 -121.31

[0320] n: Number of conformational isomers, E min The lowest energy conformational isomer, E max Highest energy conformational isomer

[0321] Leflunomide activates AhR (scheme 7), but its active metabolite A771726 does not. SGA 360 is a SAhRM that exhibits in vivo anti-inflammatory properties.

[0322] Option 7

[0323]

[0324] Example 8: AhR Combination

[0325] Peripheral blood mononuclear cells (PBMCs) were isolated from 5 mL heparinized blood samples (from 3 healthy subjects) by density gradient centrifugation (Lymphoprep; Nycomed Pharma). The PBMCs were centrifuged at 1 × 10⁻⁶. 6 Cells were resuspended at a concentration of 10% fetal bovine serum in RPMI 1640 medium supplemented with 10% fetal bovine serum and cultured in 24-well Falcon Plastic plates. The cells were stimulated with or without 100, 200, or 400 nM of AhR-binding compounds (02, 04, 13, 15, 17, or 24), dimethyl sulfoxide, or 6-carboxylic acid indodo[3,2-b]carbazole (Ficz, 200 nM) using culture medium alone or with anti-CD3 / anti-CD28 antibody (Miltenyi Biotec). Total RNA was extracted from the cells after 24 hours of culture. Cells were lysed in 1 mL of guanidine thiocyanate buffer and RNA was prepared by phenol / chloroform extraction using TRizol reagent (Invitrigen). The obtained samples were quantified by absorbance at 260 nm, and complementary DNA (cDNA) was synthesized from 1 mg of total RNA. cDNA was amplified under the following conditions: denaturation at 95°C for 1 minute; annealing at 58°C (for IFN-γ) or 60°C (for β-actin) for 30 seconds; followed by extension at 72°C for 30 seconds. Primer sequences were as follows: human IFN-γ, forward primer (SEQ ID NO.1) 5′-TGGAGACCATCAAGGAAGAC-3′, reverse primer (SEQ ID NO.2) 5′-GCGTTGGACATTCAAGTCAG-3′. IL-22 levels were assessed using commercially available TaqMan probes (Applied Biosystems). β-actin (forward primer (SEQ ID NO.3) 5′-AAGATGACCCAGATCATGTTTGAGACC-3′, reverse primer (SEQ ID NO.4) 5′-AGCCAGTCCAGACGCAGGAT-3′) was used as a housekeeping gene. Gene expression was calculated using the ΔΔCt algorithm.

[0326] Example 9: Aryl hydrocarbon receptor-driven signaling inhibits collagen synthesis in the gut

[0327] Using in vitro and in vivo fibrosis models, we have identified AhR as a modulator of collagen synthesis in the gut, as described below.

[0328] Materials and methods

[0329] Patients and samples

[0330] Mucosal samples were obtained from surgical specimens of 10 patients with fibrous stenosis colitis (CD) (median age, 37 years; range: 27–56 years); 7 of these patients were receiving corticosteroids, and the remaining patients had received corticosteroids and azathioprine. Mucosal samples were also obtained from 3 patients with ulcerative colitis (UC) who underwent colectomy for a medically unresponsive chronic disease, and 6 UC patients who underwent endoscopy for recent flare-ups (median age, 38 years; range: 29–55 years). Four UC patients were receiving corticosteroids, while the rest had received mesalazine. Normal controls included samples from 4 patients with irritable bowel syndrome and 6 patients who underwent colectomy for colon cancer, showing unaffected areas as observed macroscopically and microscopically (median age, 49 years; range: 33–68 years).

[0331] Isolation and culture of intestinal fibroblasts

[0332] Unless otherwise specified, all reagents were purchased from Sigma-Aldrich (Milan, Italy). Intestinal fibroblasts were isolated and phenotypically characterized as described elsewhere. Fibroblasts between passage 3 and 8 were used in all experiments. To examine whether AhR regulates collagen production, fibroblasts isolated from CD patients were starved overnight and then stimulated for 24–48 hours with or without Ficz (final concentration 100–400 nM; Alexis, Milan, Italy) or 2-methyl-2H-pyrazole-3-carboxylic acid (CH223191; final concentration 10 μM; Calbiochem, Nottingham, England; an AhR antagonist) with TGF-β1 (TGF-β; 1 ng / mL; Peprotech EC, London, UK) or TNF-α (15 ng / mL; R&D Systems, Abingdon, UK). Finally, RNA was extracted from the cells, and the collagen content in the cell-free supernatant was analyzed.

[0333] Induction of colonic fibrosis

[0334] Trinitrobenzenesulfonic acid (TNBS) was dissolved in 45% ethanol and administered rectally to 8-week-old female balb / c mice for 7 weeks, as previously described. Ficz (1 μg / mouse) or an AhR antagonist (CH223191; 10 μg / mouse) was dissolved in phosphate-buffered saline (PBS) and administered intraperitoneally every 48 hours after week 5 of TNBS administration. Control mice received PBS only. Signs of colitis, including weight loss, were examined in mice three weeks weekly, and mice were euthanized at week 8; tissues were then collected for histological, RNA, and collagen analysis. Colonic sections were stained with H&E and Masson trichrome to detect connective deposits. Fibrosis was scored as mild, moderate, or severe as previously reported.

[0335] RNA extraction, complementary DNA preparation and real-time polymerase chain reaction

[0336] RNA isolation, reverse transcription, and real-time PCR were performed as described above. RNA was extracted using TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer's instructions. A constant amount of RNA (1 μg per sample) was reverse transcribed into complementary DNA and amplified under the following conditions: denaturation at 95°C for 1 min; annealing at 60°C (for human collagen I (Col1A1), human Col3A1, human α-smooth muscle actin (α-SMA), and β-actin) or 58°C (for human AhR and mouse Col1A2) for 30 sec, followed by extension at 72°C for 30 sec. The primer sequences are as follows: Human Col1A1 (SEQ ID NO.5) 5′-GGACACAGAGGTTTCAGTGG-3′, (SEQ ID NO.5) 3′-GGTGACTTTGGAGACACAGG-5′; Col3A1 (SEQ ID NO.6) 5′-GGAGAATGTTGTGCAGTTTGC-3′, (SEQ ID NO.6) 3′-CGTTTGACGTGTTGTAAGAGG-5′; Human α-SMA (SEQ ID NO.7) 5′-TCTGGAGATGGTGTCACCCA-3′, (SEQ ID NO.7) 3′-ACCCACTGTGGTAGAGGTCT-5′; Human AhR (SEQ ID NO.8) 5′-GAGCACAAATCAGAGACTGG-3′, (SEQ ID NO.9) 5′-TGGAGGAAGCATAGAAGACC-3′; Mouse Col1A2 (SEQ ID NO.5) 5′-GGACACAGAGGTTTCAGTGG-3′; Mouse Col1A2 (SEQ ID NO.5) 5′-GGACACAGAGGTTTCAGTGG-3′; Mouse Col1A2 (SEQ ID NO.5) 5′-GGACACAGAGGTTTCAGTGG-3′; Mouse Col1A1 ... (SEQ ID NO.10) 5′-ACACAGTGGTATGGATGGAC-3′, (SEQ ID NO.10) 3′-CAGGTAGGTATGGTGACACA-5′; β-actin ((SEQ ID NO.3) 5′-AAGATGACCCAGATCATGTTTGAGACC-3′, (SEQ ID NO.4) 5′-AGCCAGTCCAGACGCAGGAT-3′) were used as housekeeping genes. Gene expression was calculated using the ΔΔCt algorithm.

[0337] Flow cytometry

[0338] To assess intracellular expression of AhR and phosphorylated (p) forms of p38, Erk1 / 2, NF-κB / p65, and Smad2 / 3, cells were fixed with 1% formaldehyde for 20 min, then perforated with 0.5% saponin in 1% bovine serum albumin, and treated with anti-AhR (final dilution 1:50; Abcam, Cambridge, UK), anti-p-p38 (pT180 / pY182)-PE (final dilution 1:50; BD Biosciences, San Jose, CA), anti-p-ERK1 / 2 (pT202 / pY204; pT184 / pY186)-PE (final dilution 1:50; BD Biosciences), anti-p-NF-κB / p65-FITC (final dilution 1:50; eBioscience, San Diego, CA), and anti-p-Smad2 / 3 (final dilution 1:50; Cell). Staining was performed using Signaling, Danvers, MA. Appropriate secondary antibodies and allotype-matched controls were included in all experiments (BD Biosciences). Cells were analyzed using a FACS Verseflow flow cytometer and FACS Suite software (BD Biosciences).

[0339] Collagen assay

[0340] The SircolCollagen Assay Kit (Biocolor Ltd, Belfast, UK) was used to measure fibroblast-free supernatant and mouse tissue samples according to the manufacturer's instructions.

[0341] Statistical analysis

[0342] Use Student's t-test to compare differences between groups.

[0343] result

[0344] AhR activation negatively regulates collagen production in intestinal fibroblasts.

[0345] AhR RNA transcripts were constitutively expressed in fibroblasts isolated from the intestines of patients with CD, UC, and normal controls, with no significant differences between groups (Fig. 25(A)). Flow cytometry analysis showed that nearly 50% of intestinal fibroblasts expressed AhR in both IBD and controls (Fig. 25(B)). To determine whether AhR activation regulates collagen production, fibroblasts isolated from FS of CD patients were treated with TGF-β1 or TNF-α (two known collagen inducers) for 24 hours, with or without Ficz. As expected, stimulation of fibroblasts with TGF-β1 or TNF-α induced a significant increase in the transcripts of Col1A1, Col3A1, and α-SMA (markers of fibroblast activation). Figures 26(A)-26(B) Treatment of fibroblasts with Ficz did not alter the basal RNA expression of Col1A1, Col3A1, and α-SMA, but significantly reduced the RNA transcripts of Col1A1, Col3A1, and α-SMA driven by TGF-β1 or TNF-α. Figures 26(A)-26(B) To further evaluate the role of AhR in controlling collagen expression, CD fibroblasts were stimulated with TGF-β1 or TNF-α with or without CH223191. CH223191 significantly enhanced Col1A1 RNA transcripts in unstimulated fibroblasts and significantly enhanced the RNA expression of Col1A1, Col1A3, and α-SMA in fibroblasts stimulated with TGF-β1 or TNF-α. Figures 27(A)-27(B) Analysis of soluble collagen in fibroblast culture supernatants confirmed that CH223191 significantly upregulated collagen secretion in unstimulated cells, while Ficz did not (Fig. 28(A)-28(D)). Furthermore, Ficz dose-dependently inhibited TGF-β1 and TNF-α-induced collagen secretion, while CH223191 inhibited this synthesis (Fig. 28(A)-28(D)). Neither Ficz nor CH223191 altered fibroblast viability or proliferation (data not shown).

[0346] AhR controls the activation of Map kinase in CD fibroblasts.

[0347] Activation of p38 and ERK1 / 2 MAP kinases is involved in TGF-β1 and TNF-α-driven collagen induction. Therefore, we then investigated whether AhR-mediated control of collagen synthesis is associated with changes in this intracellular pathway. To this end, we monitored p38 and ERK1 / 2 activation using flow cytometry with specific antibodies that recognize the phosphorylated / active forms of these proteins.

[0348] Under unstimulated conditions, cells expressing p-p38 or p-ERK1 / 2 were partially unaffected by Ficz, but significantly increased by CH223191. Figures 29(A)-29(B) TGF-β1 and TNF-α significantly increased the percentage of fibroblasts expressing p-p38 and p-ERK1 / 2, and this effect was decreased or increased by Ficz or CH223191, respectively. Figures 29(A)-29(B) TGF-β1 and TNF-α also enhanced the fractions of cells expressing p-Smad2 / 3 or NF-κB / p65, respectively, but Ficz and CH223191 did not change these percentages. Figures 29(A)-29(B) ).

[0349] AhR controls TNBS-induced intestinal fibrosis in mice.

[0350] To interpret these data in vivo, we used an experimental model of intestinal fibrosis induced in Balb / c mice by repeated intrarectal administration of low-dose TNBS. To determine whether AhR activation interfered with collagen synthesis and fibrosis development, mice were administered Ficz or CH223191 intraperitoneally five weeks after TNBS administration (Figures 30(A)-30(B)). Previous studies have shown that collagen deposition begins at week 4 after the first TNBS administration, and this time point was chosen based on the above results. The degree and severity of inflammation and fibrosis were assessed in animals sacrificed at week 8. As expected, mice treated with repeated doses of TNBS exhibited minimal intestinal inflammation, but significant thickening of the colonic wall. Masson's trichrome staining of colon sections and collagen RNA, and protein analysis using whole colon samples, confirmed increased collagen induction in TNBS-treated mice compared to controls. Compared with TNBS-treated mice, Ficz-treated mice showed a significant reduction in collagen expression, while mice receiving CH223191 produced more collagen (Fig. 30(B)-30(C)).

[0351] discuss

[0352] This study investigated the role of AhR in controlling intestinal fibrosis. AhR was constitutively expressed in intestinal fibroblasts isolated from FS cells of CD patients, as well as from intestinal fibroblasts isolated from UC patients and normal controls. Although treatment of CD fibroblasts with Ficz did not alter the baseline expression of collagen, inhibition of AhR with CH223191 led to increased collagen production, suggesting that constitutive AhR activation in these cell types is essential for maintaining regulated collagen synthesis. Fibroblasts isolated from the FS site in CD showed enhanced responsiveness to pro-fibrotic cytokines by producing collagen. Since studies in other systems have shown that AhR negatively regulates intracellular pathways activated by pro-fibrotic cytokines, the next step was to evaluate whether AhR activation is involved in TGF-β and TNF-α-induced collagen production. CD fibroblasts exhibited different abilities to synthesize collagen when treated with Ficz or CH223191. Specifically, Ficz dose-dependently reduced collagen RNA and protein expression, while inhibition of AhR was followed by enhanced collagen production in response to TGF-β and TNF-α. Interestingly, even at the maximum dose used in our system, Ficz did not completely eliminate cytokine-induced collagen synthesis, increasing the likelihood that AhR does not control all cytokine-driven intracellular pathways leading to collagen production. In fact, analysis of these signals revealed that Ficz completely eliminated p38 and ERK1 / 2 activation without affecting Smad2 / 3 and NF-κB activation in fibroblasts stimulated with TGF-β or TNF-α, respectively. The fact that the complete elimination of Ficz-mediated p38 and ERK1 / 2 activation was accompanied by a 60% reduction in cytokine-driven collagen synthesis suggests that MAP kinases play a major role in the control of collagen production in CD fibroblasts. Notably, Ficz and CH223191 showed no effect on Smad2 / 3 and NF-κB activation in response to TGF-β or TNF-α. These findings suggest that AhR activation does not induce a globally unresponsive state in intestinal fibroblasts, which may explain why Ficz or CH223191 does not affect the proliferation and survival of these cells.

[0353] In vivo studies in mice confirmed AhR-mediated negative regulation of collagen production, demonstrating that Ficz is effective in minimizing fibrosis associated with chronic, long-term inflammation. In contrast, mice receiving CH223191 showed stronger collagen deposition compared to control mice. In these studies, treatment with both Ficz and CH223191 began at a time point (week 5), characterized by pathological collagen accumulation in the colon of TNBS-treated mice. Therefore, AhR-mediated inhibition of collagen production is unlikely to be secondary to suppression of ongoing colitis.

[0354] In summary, these results show that AhR activation negatively controls collagen synthesis in the gut. These new findings suggest that AhR-related compounds may help prevent and / or restore FS in CD patients.

[0355] References merged

[0356] For all purposes, the full disclosure of each patent document and scientific literature mentioned herein is incorporated herein by reference.

[0357] equivalent

[0358] This invention may be embodied in other specific forms without departing from its essential characteristics. Therefore, the foregoing embodiments are to be considered illustrative and not limiting of the invention described herein. The scope of the invention is indicated by the appended claims, not by the foregoing description, and all changes falling within the meaning and scope of the equivalents of the claims are intended to be included therein.

Claims

1. Compounds represented by the following formula: ， Or its medication may be acceptable with salt.

2. A pharmaceutical composition comprising the compound of claim 1; and a pharmaceutically acceptable carrier.

3. Use of a therapeutically effective amount of the compound according to claim 1 in the preparation of a medicament for treating arthritis, wherein the arthritis is selected from psoriatic arthritis, rheumatoid arthritis, and juvenile arthritis.

4. The use of claim 3, wherein the juvenile arthritis is juvenile rheumatoid arthritis.

5. Use of a therapeutically effective amount of the compound according to claim 1 in the preparation of a medicament for treating Crohn's disease.

6. The use of claim 5, wherein the Crohn's disease is selected from ileocolic inflammation, ileitis, and gastroduodenal Crohn's disease.

7. The use of claim 6, wherein the ileocolitis is granulomatous ileocolitis.

8. The use of claim 6, wherein the ileitis is jejunostomy.

9. The use of claim 5, wherein the Crohn's disease is intestinal fibrosis.

10. The use of claim 5, wherein the Crohn's disease is fibrostenotic Crohn's disease.

11. Use of a therapeutically effective amount of the compound of claim 1 in the preparation of a medicament for the prevention, treatment or reduction of fibrotic stenosis or colonic fibrosis in a subject.

12. Use of claim 11, wherein the fibrous stricture or colonic fibrosis is associated with Crohn's disease.

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