Novel antifibrotic drugs

Abstract

The present invention relates to novel cinnamic acid amides that can be used in the treatment of fibrosis and neoplasms, and cinnamic acid amides for use in the treatment of fibrosis, neoplasms, arthrolithiasis, familial Mediterranean fever and pericarditis.Furthermore, the present invention relates to pharmaceutical compositions comprising said cinnamic acid amides, and screening assays for identifying compounds suitable for the treatment of fibrosis.

Description

[Technical field]

[0001] TECHNICAL FIELD OF THEINVENTION The present invention relates to novel cinnamic acid amides that can be used in the treatment of fibrosis and neoplasms, and cinnamic acid amides for use in the treatment of fibrosis, neoplasms, arthrolithiasis, familial Mediterranean fever and pericarditis.Furthermore, the present invention relates to pharmaceutical compositions comprising said cinnamic acid amides, and screening assays for identifying compounds suitable for the treatment of fibrosis. [Background technology]

[0002] Fibrotic diseases affect nearly every tissue in the body and account for more than 45% of all deaths in industrialized countries, with progressive disease rapidly leading to organ damage, organ failure, and eventual death (1-3). Due to its ubiquitous presence and high mortality rate, there is a growing medical need for novel drug discovery strategies against fibrosis, or “scarring” (3, 4). However, no effective antifibrotic agents exist in the clinic. The lack of antifibrotic therapies and the associated high medical need are best exemplified by idiopathic pulmonary fibrosis (IPF), a rapidly progressive and fatal fibrotic disorder. Patients with this common form of interstitial fibrotic lung disease have a median survival of 3-5 years (5-7). Currently, there are only two approved antifibrotic drugs on the market for IPF, pirfenidone and nintedanib, but both substances partially slow the rate of decline in lung function but do not halt disease progression (8-10). Thus, novel treatment strategies and approaches are urgently needed. In the pathogenesis of fibrosis, repeated and continuous injury leads to persistent and self-proliferating activation of fibroblasts, resulting in their transdifferentiation into synthetic, highly contractile α-smooth muscle actin (αSMA)-expressing myofibroblasts that deposit large amounts of extracellular matrix (ECM), causing lung stiffening and disrupting normal lung architecture (3, 6, 11, 12). Matrisomes in the fibrotic ECM have been shown to harbor disease- and progression-specific signatures of fibrillar collagens (types I, III, and V), proteoglycans, fibronectin, glycosaminoglycans, matrix Gla protein, and microfibril-associated proteins (11, 13-16).

[0003] Of all the reported profibrotic signals, the multifunctional TGF-β1 is the most intensively studied and central in various fibrotic diseases where it can trigger the transdifferentiation of fibroblasts into myofibroblasts (17-21). TGF-β1 binds to its TGF-β1 receptor, and downstream signaling occurs through post-translational modification of cytoplasmic members of the SMAD family, which function as transcription factors in the cell nucleus and regulate the expression of common profibrotic genes, including ECM proteins (22-25). Plasminogen activator inhibitor-1 (PAI-1), an essential downstream target of the TGF-β1 pathway, suppresses the fibrinolytic system and is considered as a therapeutic target of choice for fibrosis (26). Moreover, in IPF, profibrotic IL8 has recently been found to be secreted by a specialized population of fibrogenic mesenchymal progenitor cells with autocrine effects on proliferation and motility as well as paracrine effects on macrophage recruitment (27).

[0004] Tranilast is known as a mast cell degranulation inhibitor developed by Kissei Pharmaceutical Co., Ltd., and has already been approved in Japan and Korea in 1982 for the treatment of bronchial asthma, keloids and hypertrophic scars. The drug appears to act by inhibiting the release of histamine from mast cells, but its molecular target remains unclear. The antifibrotic properties of tranilast have also been reported in the prior art, but its efficacy is very low (IC 50 The therapeutic efficacy of tranilast is approximately 150 μM, but high doses may be required in humans, which have been reported to cause liver toxicity. Thus far, medicinal chemistry optimization efforts have failed to significantly improve the antifibrotic activity of tranilast (28).

[0005] Thus, there is a general need for antifibrotic drugs and assays for the identification of suitable antifibrotic drugs. Tranilast may be a suitable lead compound for further medicinal chemistry optimization. Summary of the Invention

[0006] The present invention is directed to compounds for use in the treatment of fibrosis and neoplasms, preferably fibrosis or neoplasms located in the heart, lungs, renal tract, liver, skin, pleura and retroperitoneum, more preferably, the fibrosis is selected from pleural fibrosis, retroperitoneal fibrosis, atrial fibrillation, myocardial interstitial fibrosis, idiopathic pulmonary fibrosis (IPF), interstitial lung disease, chronic kidney disease, non-alcoholic fatty liver disease, skin scars, keloids, tumor-associated fibrotic responses, wherein the compound has the formula (I): A compound according to TIFF2024527473000001.tif42128, During the ceremony R 1 -OR 12 , -O(CH2) u (C3~C 10 ) Aryl, -O(CH2) u (C3~C 10 ) Cycloalkyl, -O(CH2) u (C2) Alkynyl; -(CH2) u (C3~C 10 )Aryl, -O(CH2) u (C3~C 10 ) cycloalkyl, -(CH2) u (C3~C 10 ) cycloalkyl, TIFF2024527473000002.tif15128, -(CH2) u (C2) alkynyl; u is 0 to 6; R 2 ~R 5 are independently H, -OR 12 , -(C1~C 10 )Alkyl, halogen, cyano, isocyano, cyanato, isocyanato, thiocyanato, isothiocyanato, azido, -(C2-C 10 ) alkenyl, -(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 )Heterocyclyl, -(C3~C 10 )Aryl, -(C3~C 10)heteroaryl, -CHZ2, -CZ3-CH2Z, -OCHZ2, -OCZ3, -OCH2Z- -N(R 13 )(R 14 ), -N(R 15 )(OR 16 ), -S(O) 0~2 R 17 , -S(O) 1~2 OR 18 , -OS(O) 1~2 R 19 , -OS(O) 1~2 OR 20 , -S(O) 1~2 N(R 21 )(R 22 ), -OS(O) 1~2 N(R 23 )(R 24 ), -N(R 25 )S(O) 1~2 R 26 , -NR 27 S(O) 1~2 OR 28 , -NR 29 S(O) 1~2 N(R 30 )(R 31 ), -C(=X)R 32 , -C(=X)XR 33 , -XC(=X)R 34 , and -XC(=X)XR 35 , -OR 36 , -O(CH2) v (C3~C 10 )Aryl, -O(CH2) v (C3~C 10 ) Cycloalkyl, -O(CH2) v (C2) alkynyl; R 6 H, -(C1~C 10 ) alkyl, benzyl and -(CH2) 1~5 (C3~C 10 )cycloalkyl; where -(C1-C 10 ) alkyl, benzyl and -(CH2) 1~5 (C3~C 10) cycloalkyl may be further substituted with at least one substituent selected from the group consisting of halogen, preferably F; R 7 ~R 11 are independently H, -OR 12 , -SR 12 , -(C1~C 10 )Alkyl, halogen, -(C1-C 10 )AlkylO(C1-C 10 )Alkyl, cyano, isocyano, cyanato, isocyanato, thiocyanato, isothiocyanato, azido, -(C2-C 10 ) alkenyl, -(C2-C 10 ) alkynyl, -O(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 )Heterocyclyl, -(C3~C 10 )Aryl, -(C3~C 10 ) Heteroaryl, -(CH2) v CHZ2, -CZ3-CH2Z, -OCHZ2, -OCZ3, -OCH2Z, -N(R 13 )(R 14 ), -N(R 15 )(OR 16 ), -S(O) 0~2 R 17 , -S(O) 1~2 OR 18 , -OS(O) 1~2 R 19 , -OS(O) 1~2 OR 20 , -S(O) 1~2 N(R 21 )(R 22 ), -OS(O) 1~2 N(R 23 )(R 24 ), -N(R 25 )S(O) 1~2 R 26 , -NR 27 S(O) 1~2 OR 28 , -NR 29 S(O) 1~2 N(R 30 )(R 31 ), -C(=X)R32 , -C(=X)XR 33 , -XC(=X)R 34 , and -XC(=X)XR 35 , -O(CH2) v (C3~C 10 ) Cycloalkyl, -O(CH2) v (C1~C 10 ) alkyl and -O(CH2) v (C3~C 10 ) aryl; Here, R 1 ~R 5 and R 7 ~R 11 The two adjacent remaining moieties of the formula (III) to (XI): Optionally, a ring may be formed attached to the basic aromatic ring of formula (I) according to TIFF2024527473000003.tif26160; In the formula, T 1 and T 2 are independently H, -(C1 to C 10 ) selected from the group consisting of alkyl and halogen; Here, each hydrogen in the formulas (III) to (IX) is a halogen or -(C 10 ) optionally substituted with aryl, -(C1-C3)alkyl, preferably F; Het is O, S, NH, N(C1-C 10 ) alkyl; G is selected from CH and N; J1 to J4 are independently selected from C or N, preferably J1 to J4 are C; Here, when any one of J1 to J4 is N, the corresponding R bonded to each of J1 to J4 which is N 1 ~R 4 does not exist; R 12 ~R 36 are independently H, -(C1 to C 10 )Alkyl, -(C2-C 10 ) alkenyl, -(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C10 )Heterocyclyl, -(C3~C 10 )Aryl, -(C3~C 10 ) heteroaryl; R 38 are independently H, -(C1 to C 10 ) alkyl; Independently - (C1~C 10 )Alkyl, -(C2-C 10 ) alkenyl, -(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 )Heterocyclyl, -(C3~C 10 )Aryl, -O(CH2) v (C3~C 10 ) Cycloalkyl, -O(CH2) v (C1~C 10 ) alkyl and -O(CH2) v (C3~C 10 ) R selected from the group consisting of aryl 1 ~R 11 , and R 12 ~R 35 OR 12 , -(C1~C 10 )Alkyl, halogen, cyano, isocyano, cyanato, isocyanato, thiocyanato, isothiocyanato, azido, -(C2-C 10 ) alkenyl, -(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 )Heterocyclyl, -(C3~C 10 )Aryl, -(C3-C 10 )heteroaryl, -CHZ2, -CZ3-CH2Z, -OCHZ2, -OCZ3, -OCH2Z-, -N(R 13 )(R 14 ), -N(R 15 )(OR 16 ), -NHC(O)(C1-C 10 ) alkyl, -S(O) 0~2 R 17 , -S(O) 1~2 OR 18, -OS(O) 1~2 R 19 , -OS(O) 1~2 OR 20 , -S(O) 1~2 N(R 21 )(R 22 ), -OS(O) 1~2 N(R 23 )(R 24 ), -N(R 25 )S(O) 1~2 R 26 , -NR 27 S(O) 1~2 OR 28 , -NR 29 S(O) 1~2 N(R 30 )(R 31 ), -C(=X)R 32 , -C(=X)XR 33 , -XC(=X)R 34 , and -XC(=X)XR 35 , -OR 36 , and -O(CH2) v (C3~C 10 ) optionally further substituted with at least one substituent selected from the group consisting of aryl; v is 0 to 5; Z is a halogen; X is selected from the group consisting of O, -NH- or S; A is TIFF2024527473000004.tif36128; n is 1, 2, or 3, preferably 1; o is 1, 2, or 3, preferably 1; R is H, (C1-C6) alkyl, cyano, -(C3-C 10 ) cycloalkyl, benzyl, or R is TIFF2024527473000005.tif15128 by R 7 or R 11 is part of a ring connecting R 37 is H or -CF3; However, if n is 2 or 3, A is Could be TIFF2024527473000006.tif15128; However, R 5 is not -COOH.

[0007] The present invention further relates to a compound according to formula (II) TIFF2024527473000007.tif47128 expression R 1 -OR 12 , -O(C5~C 10 ) Heteroaryl, -O(C3-C 10 )Aryl, -O(CH2) u (C3~C 10 ) cycloalkyl, TIFF2024527473000008.tif14128, -O(CH2) u (C3~C 10 ) aryl; R 2 ~R 5 and R 7 , and R 11 are independently H, -(C1 to C 10 ) Alkyl, halogen, azido, cyano, -O(C1-C 10 ) alkyl, -(CH2) u (C3~C 10 ) Aryl, -(CH2) u (C3~C 10 )Cycloalkyl, -(C2-C 10 ) alkenyl, -(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 ) aryl, which may be further substituted with at least one substituent selected from the group consisting of halogen, -OH, -NH2, -NHC(O)CH3, -CN, -N3, and -COOH, -C(O)NH2; R 6 is H, -(C1~C 10 ) alkyl, benzyl and -(CH2) 1~5 (C3~C10 ) cycloalkyl; 10 ) alkyl, benzyl and -(CH2) 1~5 (C3~C 10 ) cycloalkyl may be further substituted with at least one substituent selected from the group consisting of halogen, preferably F; R 8 , R 9 and H, -O(C1-C 10 ) alkyl, -SR 12 , -O(CH2) u (C3~C 10 )Aryl, -O(CH2) u (C3~C 10 )Cycloalkyl, -O(C3-C 10 ) cycloalkyl, or -O(C2-C 10 ) alkenyl; R 10 is H, halogen, -O(C1-C 10 ) alkyl, -O(CH2) u (C3~C 10 )Aryl, -O(CH2) u (C3~C 10 )Cycloalkyl, -O(C3-C 10 ) cycloalkyl, or -O(C2-C 10 ) alkenyl; However, if R9=H, R8 or R 10 -OR 12 and; u is 0 to 6; R is H, (C1-C6) alkyl, cyano, -(C3-C 10 ) cycloalkyl, benzyl, or R is TIFF2024527473000009.tif14128 by R 7 or R 11 is part of a ring connecting R 37 is H or -CF3; R 12 are independently H, -(C1 to C 10)Alkyl, -(C2-C 10 ) alkenyl, -(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 )Heterocyclyl, -(C3~C 10 )Aryl, -(C3~C 10 )heteroaryl, (CH2) u (C3~C 10 ) Aryl, -(CH2) u (C3~C 10 ) Heteroaryl, -(CH2) u (C3~C 10 )cycloalkyl; preferably -(C1-C 10 )alkyl, more preferably -(C1-C4)alkyl; Here, R 8 ~R 10 The two adjacent remaining parts of Optionally, a ring may be formed attached to the basic aromatic ring of formula (II) according to TIFF2024527473000010.tif27157; In the formula, T 1 and T 2 are independently H, -(C1 to C 10 ) selected from the group consisting of alkyl and halogen; Here, each hydrogen in the formulas (III) to (XI) is a halogen, -(C 10 ) optionally substituted with -(C1-C3)aryl, or -(C1-C3)alkyl, preferably F; Het is O, S, N (C1-C 10 ) alkyl or NH; preferably O; R 38 are independently H, -(C1 to C 10 ) alkyl; G is selected from CH, N; J1 to J4 are independently selected from C or N, preferably J1 to J4 are C; Here, when any one of J1 to J4 is N, the corresponding R bonded to each of J1 to J4 which is N 1 ~R 4does not exist; however, When J1 to J4 are C, and I)R 5 is -(CH2)3CH3; R 9 is -OCH3, -OCH2CH3, -O(CH2)2CH3, -OCH2phenyl or -O(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R is H; and (a)R 8 and R 10 is H; or (b)R 8 is -OCH3 or -OCH2CH3, and R 10 is H; or (c)R 10 is -OCH3 or -OCH2CH3, and R 8 is H Case;R 6 is not H; II) R 9 is -OCH3, -O(CH2)2CH3, -O(2-propyl), -O(CH2)4CH3, -O(CH2)5CH3, -OCH2(4-chlorophenyl), -O(CH2)2CH(CH3)2, -OCH2(2,6-dichlorophenyl), or -OCH2phenyl; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R is H; R 8 is -OCH3 and R 10 is H or R 10 is -OCH3 and R 8 is H;R 5 is -(CH2)3CH3 Case;R 6 is not H; III) R 9 is -OCH3; R 8 is Br and R 10is H or R 10 is Br and R 8 is H;R 5 is -(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 11 and R is H In this case, R 1 is not H; IV) R 5 is -(CH2)3CH3, and R 9 is -OCH3, R 7 is -OCH3 and R 11 is H or R 11 is -OCH3 and R 7 is H;R 1 , R 2 , R 3 , R 4 , R 8 , R 10 , R 11 and R is H Case;R 6 is not H; V)R 9 is -OCH3, -OCH2phenyl, or -OCH2(2-fluorophenyl); R 8 is Br and R 10 is -OCH3 or R 10 is Br and R 8 is -OCH3; R 5 is -O(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R is H In this case, R 6 is not H; VI)R 9 is -O(CH2)3CH3; R 8 is -OCH2CH3 and R 10 is H or R 10 is -OCH2CH3 and R 8 is H;R5 is -O(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R is H In this case, R 6 is not H; VII) R 9 is -OCH2 (2-chlorophenyl); R 8 is Br and R 10 is -CH2CH3 or R 10 is Br and R 8 is -OCH2CH3; R 5 is -(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R is H In this case, R 6 is not H; VIII)R 9 is -O(2-octenyl); R 8 is Cl and R 10 is H or R 10 is Cl and R 8 is H;R 5 is -(CH2)3COOH; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R is H In this case, instead of H; IX)R 9 is -OCH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 8 , R 10 , R 11 , R is H; and R 5 is -(2-fluorophenyl), -phenyl Case;R6 is not H; X)R 9 is -OCH2CH3; R 5 is -(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 8 , R 10 , R 11 and R is H In this case, R 6 is not H; XI)R 9 is -OCH3; R 8 is -OCH3 and R 10 is H or R 10 is -OCH3 and R 8 is H;R 5 is -(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 8 , R 7 and R is H In this case, R 6 is not H; XII) R 9 is -OCH3; R 5 is -O(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R is H and R 8 is -OCH2CH3 and R 10 is H or R 10 is -OCH2CH3 and R 8 is H In this case, R 6 is not H; XIII)R 5 is -(CH2)3CH3; R 9 is -O(CH2)3CH3 or -OCH3; R 1 , R 2 , R 3 , R4 , R 7 , R 8 , R 10 , R 11 and R is H In this case, R 6 is not H; XIV)R 5 is -CH3, -(CH2)2CH3 or -CH2CH3; R 9 is -OCH3; R 8 is -OCH3 and R 10 is H or R 8 is H and R 10 is -OCH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R is H Case;R 6 is not H; XV)R 5 is -(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 8 , R 9 , R 10 and R 11 is H Case;R 6 is not H.

[0008] In a further aspect, the present invention relates to a pharmaceutical composition comprising a compound as defined above.

[0009] In a further aspect, the present invention is directed to compounds and pharmaceutical compositions as defined above for use in medicine, in particular for use in the treatment of fibrosis and neoplasms, preferably fibrosis or neoplasms located in the heart, lungs, renal tract, liver, skin, pleura and retroperitoneum, more preferably, the fibrosis is selected from pleural fibrosis, retroperitoneal fibrosis, atrial fibrillation, myocardial interstitial fibrosis, idiopathic pulmonary fibrosis (IPF), interstitial lung disease, chronic kidney disease, non-alcoholic fatty liver disease, skin scars, keloids, tumor-associated fibrotic reactions.

[0010] In a further aspect, the present invention is directed to compounds as defined above for use in medicine, in particular for use in the treatment of inflammatory diseases such as arthritic disease, familial Mediterranean fever and pericarditis, and to pharmaceutical compositions.

[0011] Further, the present invention relates to a method for producing a pharmaceutical composition comprising the steps of: (a) culturing adherent cells that deposit at least one protein in the presence of at least one test compound; (b) staining of at least one protein deposited by adherent cells; (c) fixation of the adherent cells and the at least one protein; (d) microscopic detection of the at least one stained deposited protein signal; (e) data analysis of the signals detected in step (d), comprising quantifying the amount of said at least one protein deposited in the presence of said at least one test compound. wherein step (b) is performed before step (c).

[0012] In dose-response studies, the compounds of the present invention proved to be >100-fold more potent in inhibiting ECM deposition compared to tranilast when applying the assay of the present invention (see Figures 14A-B). The compounds of the present invention were found to exhibit dynamic inhibition of ECM deposition without cell death (see Figure 14E). Unlike tranilast, exposure of patient-derived primary human lung fibroblasts (phLF) to the compounds of the present invention was found to switch cell morphology from elongated to round cells, including extensive reorganization of the actin cytoskeleton (see Figure 14F). These substantial morphological changes observed with the compounds of the present invention but not with tranilast strongly indicate a distinct mode of action (see Figure 12H).

[0013] For the assay of the present invention, it has been found that performing immunostaining according to step (b) before fixation according to step (c) identifies only proteins deposited outside the cells and does not result in "false positive" hits due to staining of intracellular ECM precursor proteins. The assay can be used for the quantification of deposited ECM of any adherent cell (patient-derived primary, animal-derived primary, human cell line, animal cell line) that produces ECM, in any organ (healthy or diseased) or from various animal species and / or animal disease models. In one embodiment, human primary cells from patients (e.g., lung fibroblasts from IPF patients) are used. This produces the most clinically relevant efficacy and potency data possible in vitro, especially when compared to assays using immortalized cell lines or cells from different animal species. [Brief description of the drawings]

[0014] [Figure 1A] Figure 1: Live labeling of phLFs ensures extracellular fluorescent staining of only ECM proteins. (A) Untreated phLFs were either PFA fixed, PFA fixed or Triton permeabilized, or stained alive. Only unfixed live staining resulted in characteristic extracellular staining (white arrows) without cytoplasmic signal. Scale = 100 μm. (B) Western blot analysis of culture supernatants of phLFs treated or not with Brefeldin A demonstrated that treatment with Brefeldin A inhibited the secretion of both ECM proteins collagen I and fibronectin. (C) Confocal fluorescence microscopy of Brefeldin A-treated phLFs that were either PFA fixed, PFA fixed or Triton permeabilized, or stained alive. Immunostaining for collagen I (red) demonstrated that PFA fixation, with or without Triton permeabilization, positively labels intracellular collagen I (white arrowheads), whereas staining of live phLF characteristically labels only extracellular collagen I (white arrows). Phalloidin (green) stains intracellular actin stress fibers, and nuclei were stained with Hoechst (blue). Scale = 100 μm. [Figure 1B] See legend to Figure 1A. [Figure 1C] See legend to Figure 1A. [Figure 2A] Figure 2: Protein analysis of intracellular and secreted ECM proteins in a human 3D fibrosis model. (A) Treatment of phLF with TGFβ1 resulted in a significant increase in soluble intracellular and secreted collagen I protein. (B) Treatment of phLF with TGFβ1 resulted in a significant increase in soluble intracellular and secreted collagen V protein. (C) Co-treatment of TGFβ1 with ethyl 3,4-dihydroxybenzoate (EDHB), a prolyl-4-hydroxylase inhibitor, resulted in a statistically significant suppression of intracellular collagen I, collagen V, and surprisingly not collagen fibulin 1 expression. (D) Co-treatment of TGFβ1 with EDHB resulted in a significant suppression of secreted collagen I. ns = not significant. * p<0.05, ** p<0.01 and *** p<0.001. Statistics: one-way ANOVA with Bonferroni correction. All quantitative data represent the mean ± SEM. 3 (phLF of 3 different patients). [Figure 2B] See legend to Figure 2A. [Figure 2C] See legend to Figure 2A. [Figure 2D] See legend to Figure 2A. [Figure 3A]Figure 3: Image analysis by FANTAIL applying deep convolutional neural networks (CNNs), CNN training and hyperparameter optimization. (A) For data augmentation in the training set, each original image was fragmented into tiles of size np×np with 3 / 4 overlap and stored at 0°, 90°, 180° and 270° rotation orientations. (B) For classification of original images, each image was fragmented into non-overlapping tiles of size np×np. Each tile was classified separately as "hit" or "other" (see also Figure 3A). (C) Learning curves of deep CNNs for training accuracy depending on np (Figure 10A). (D) Learning curves of deep CNNs for validation accuracy depending on np (Figure 10A). (E) Receiver operating characteristic analysis as a performance evaluation per classification problem based on the proportion of tiles classified as hits using the deep CNNs identified in Figure 10A depending on np and the number of training iterations (= epochs). (F) Image clustering is achieved by flattening the image's pixel matrix into a linear vector and projecting it into two-dimensional space using UMAP. [Figure 3B] See legend to Figure 3A. [Figure 3C] See legend to Figure 3A. [Figure 3D] See legend to Figure 3A. [Figure 3E] See legend to Figure 3A. [Figure 3F] See legend to Figure 3A. [Figure 4A]Figure 4: Cytotoxicity of tranilast in phLF and its dose-response relationship in inhibiting ECM deposition. (A) MTT assay showing cell viability in phLF+TGFβ1 and phLF, both treated with various tranilast concentrations (75 μM, 150 μM, 300 μM). Cell death was mimicked by treating phLF with 10% EtOH. (B) Protein analysis by Western blotting demonstrating a significant dose-dependent decrease in cytosolic αSMA in phLF+TGFβ1+tranilast. (C) Protein analysis by Western blotting demonstrating a significant dose-dependent decrease in soluble fibulin 1 in phLF+TGFβ1+tranilast. (D) Protein analysis by Western blotting showing a significant dose-dependent decrease in soluble collagen I and collagen V in phLF+TGFβ1+tranilast. (E) Analysis of gene expression by qPCR showing a significant dose-dependent downregulation of αSMA and collagen I, but surprisingly not fibulin 1 and collagen V transcripts. ns = not significant. * p<0.05 and ** p<0.01. Statistics: unpaired t-test combined with Bonferroni's multiple comparison test. All quantitative data represent the mean ± SEM. n = 3 (phLF of 3 different patients). [Figure 4B] See legend to Figure 4A. [Figure 4C] See legend to Figure 4A. [Figure 4D] See legend to Figure 4A. [Figure 4E] See legend to Figure 4A. [Figure 5A]Figure 5: UMAP Regulatory Pattern Clustering (UMAP-RPC) of Transcriptome Data and Network Analysis (A-C). The 20 most highly and least highly deregulated genes of phLF+TGFβ1 (A), phLF+TGFβ1+Example 84 (B) and phLF+Example 84 (C) compared to all untreated phLF controls are shown as heatmaps of three different patient samples (n=3). (D) 279 overlapping genes (>2-fold, FDR<10%) counter-regulated in phLF+TGFβ1+Example 84 compared to phLF+TGFβ1. (E) Diagram of UMAP-RCP based on differential transcript abundance ("a,b,c,d") for different conditions (phLF, phLF+TGFβ1, phLF+TGFβ1+Example 84, phLF+Example 84) illustrated for one arbitrary gene. The normalized 4-dimensional vectors are subsequently reduced to 2-dimensional vectors by UMAP. (F) Two examples as in (A) for MMP1 abundant in condition "d" but not elsewhere and MYH abundant in condition "c" but not elsewhere. (G) UMAP clustering of all differentially expressed genes (>2-fold, FDR<10%) for similarity of gene expression patterns between the four different conditions in phLF, phLF+TGFβ1, phLF+TGFβ1+Example 84, and phLF+Example 84. Thus, green and red boxes highlight clustering of genes with similar expression, while blue and red color codes indicate low and high transcript abundance, respectively. [Figure 5B] See legend to Figure 5A. [Figure 5C] See legend to Figure 5A. [Figure 5D] See legend to Figure 5A. [Figure 5E] See legend to Figure 5A. [Figure 5F] See legend to Figure 5A. [Figure 5G] See legend to Figure 5A. [Figure 6A]Figure 6: Transcriptomic cluster and network analysis. (A) List of deregulated genes in cluster A, which were found to form an interacting protein network based on analysis in the STRING database. [Figure 6B] Figure 6: Transcriptomic cluster and network analysis. (B) List of deregulated genes in cluster B, which were found to form an interacting protein network based on the analysis in the STRING database. [Figure 7A] Figure 7: Proteomic analysis of human precision-cut lung sections (hPCLS) treated with N23P identifies upregulated pro-fibrotic target networks. (A) Heatmap showing abundance of the 20 most (red) and least (blue) deregulated proteins in fibrosis cocktail (FC)-treated PCLS as fold change in FC to CC ratio (n=3, 3 different patients: P1, P2, P3). Proteins related to tissue fibrosis are indicated with red asterisks. (B) Diagram of UMAP clustering based on abundance of differential proteins ("a, b, c") per different conditions (hPCLS, hPCLS+FC, hPCLS+FC+Example 84, and hPCLS+FC+Tranilast). (C) Based on STRING DB analysis, cluster A contained functional sub-networks involved in extracellular matrix organization (green), actin cytoskeleton (pink), and interleukin signaling (yellow). (D) List of deregulated proteins in cluster A that were found to form an interacting protein network based on analysis in the STRING database. [Figure 7B] See legend to Figure 7A. [Figure 7C] See legend to Figure 7A. [Figure 7D] See legend to Figure 7A. [Figure 8A]Figure 8: 3D assessment of ECM deposition by using primary human lung fibroblasts derived from IPF patients. (A) Primary human lung fibroblasts (phLF) are derived from explanted IPF lungs, grown in cell culture, and used for high-throughput drug screening and hit validation. (B) Clinical data of the patients from whom phLF were derived. (C) Diagram of the actual workflow used in the ECM deposition assay. (D) Software-based volume rendering of confocal Z-stacks of immunostained ECM (collagen I in red) used for quantification of ECM volume and automated cell counting (Hoechst-stained cell nuclei in blue). Scale = 500 µm. (E) Orthoview of a confocal Z-stack of phLF (red) depositing collagen V (green) only outside (indicated by white arrows) and on the surface of the cells. Cell nuclei are stained with Hoechst (blue). Scale = 50 µm and 25 µm. (F) Complex 3D ECM network of collagen I (red) and collagen V (green) stained fibers showing areas of colocalized fibers (white arrows) and single fibers (white arrowheads). Cell nuclei are stained with Hoechst (blue). Confocal Z-stacks are shown as maximum intensity projection images. Scale = 50 μm. (G) 4D confocal time-lapse imaging of phLF for 16 h showing the various dynamic processes occurring upon assembly of a single collagen I-ECM fiber (red). White arrows indicate single ECM fibers. Colored asterisks indicate single cell nuclei (blue Hoechst) of individual cells. Scale = 50 μm. [Figure 8B] See legend to Figure 8A. [Figure 8C] See legend to Figure 8A. [Figure 8D] See legend to Figure 8A. [Figure 8E] See legend to Figure 8A. [Figure 8F] See legend to Figure 8A. [Figure 8G] See legend to Figure 8A. [Figure 9-1]Figure 9: 3D fibrotic disease model using primary human lung fibroblasts derived from IPF patients. (A) phLFs treated with 1 ng / ml TGFβ1 transdifferentiated into myofibroblasts that incorporated αSMA (green) into actin stress fibers. Cell nuclei are stained by Hoechst (blue). Scale = 200 μm. Quantification of mean fluorescence intensity (MFI) of αSMA expression in phLFs of three different patients after TGFβ1 treatment (n = 3). (B) Venn diagram showing the overlap of 17 ECM proteins between the myofibroblast surface proteome (pink) and the published "core matrisome" (blue). (C) Heatmap of protein expression levels of ECM proteins on the surface of myofibroblasts (phLF+TGFβ1), identifying collagen I and fibulin 1 among the most upregulated ECM proteins. Red and blue indicate high and low protein expression levels, respectively. (D) 3D confocal immunofluorescence microscopy of phLFs and phLF+TGFβ1. phLF+TGFβ1 showed increased ECM deposition of collagen I (red), collagen V (green) and fibrin 1 (yellow). Cell nuclei were stained with Hoechst (blue). Confocal Z-stacks are shown as maximum intensity projection images. Scale = 500 μm. (E) Software-based quantification of deposited ECM volume shows a significant increase in the amount of deposited ECM in phLF+TGFβ1, while the amount of cells remains unchanged. n = 4 (phLF from 4 different patients). Statistics: one-way ANOVA with Bonferroni correction. (F) 3D confocal immunofluorescence microscopy evaluating ECM deposition in phLF, phLF+TGFβ1, and phLF+TGFβ1+EDHB. Ethyl 3,4-dihydroxybenzoate (EDHB) treatment inhibits ECM deposition of collagen I (red), collagen V (green) and fibrin 1 (yellow). Cell nuclei were stained with Hoechst (blue). Confocal Z-stacks are shown as maximum intensity projection images. Scale = 500 μm. (G) Software-based quantification of deposited ECM volume for data shown in (F). Data are presented as mean ± SEM. Differences between groups were assessed by paired t-test. * p<0.05. [Figure 9-2] See description of Figure 9-1. [Figure 9-3]See description of Figure 9-1. [Figure 10A] Figure 10: Fibrotic pattern detection by artificial intelligence using CNN for hit identification within ECM deposition screening data of 1509 FDA-approved compounds (FANTAIL). (A) Overview of the supervised multi-layer deep convolutional neural network (CNN) developed to detect fibrotic and non-fibrotic patterns within images containing deposited ECM derived from 3D confocal microscopy of immunofluorescently labeled collagen I, collagen V and fibrin 1. (B) The training dataset consisted of assay controls and additional samples treated with phLF+TGFβ1 supplemented with 5% ethanol. The CNN network was uniquely trained to detect false positive hits due to inhibitors of ECM deposition and cytotoxic effects. (C) False positive hits due to immunofluorescence artifacts were resolved by UMAP clustering of the predicted hits. After this final filtering, N-(3',4'-dimethoxycinnamoyl)-anthranilic acid (tranilast) was determined to be a promising candidate for repurposing in IPF. (D) Pie chart showing the classification of detected hits into groups of similar molecular functions and biological processes. Tranilast is classified as a GPCR target molecule. (E) 3D confocal microscopy of immunofluorescently labeled collagen I (red), collagen V (green) and fibulin 1 (yellow) to verify the inhibitory effect of tranilast-treated phLF+TGFβ1 on ECM deposition in a dose-dependent manner (untreated, vehicle, 75 μM, 150 μM, 300 μM). Cell nuclei were stained by Hoechst (blue). Confocal Z-stacks are shown as maximum intensity projection images. Scale = 500 μm. (F) Quantitative and statistical analysis of the inhibitory effect of tranilast on phLF+TGFβ1 on ECM deposition of collagen I, collagen V and fibulin 1 normalized to cell number. ns = not significant. * p<0.05 and ** p<0.01. Statistics: one-way ANOVA with Bonferroni correction. All quantitative data represent the mean ± SEM. n = 3 (phLF of 3 different patients). [Figure 10B] See legend to Figure 10A. [Figure 10C]See legend to Figure 10A. [Figure 10D] See legend to Figure 10A. [Figure 10E] See legend to Figure 10A. [Figure 10F] See legend to Figure 10A. [Figure 11-1] Figure 11: Genome-wide transcriptome analysis of N23P identifying novel anti-fibrotic target networks. (A) Experimental summary of transcriptional analysis of a human fibrosis model and treatment with ECM deposition inhibitor N23P. (B) Volcano plot showing all significantly differentially expressed genes (>2-fold, FDR<10%) in phLF+TGFβ1, phLF+TGFβ1+Example 84, and phLF+Example 84, highlighting the top 10 most (red) and least (blue) abundant transcripts. (C) Venn diagram showing 362 overlapping genes between 2076 deregulated genes in phLF+TGFβ1 and 661 deregulated genes in phLF+TGFβ1+Example 84. (D) Gene set enrichment analysis (GSEA) of phLF+TGFβ1+Example 84 showing negative enrichment of pro-fibrotic gene signatures such as collagen formation, extracellular matrix organization, and smooth muscle contraction. (E) UMAP-RPC overlaid with each gene colored as fold change in transcript abundance within its cluster. Upregulated genes are shown in red and downregulated genes are shown in blue. Boxed cluster A (red) shows genes found mostly upregulated in phLF+TGFβ1 only, boxed cluster B (green) shows genes upregulated in phLF+TGFβ1+Example 84 and phLF+Example 84. (F) Based on STRING DB analysis, cluster A contained functional sub-networks of molecular components involved in extracellular matrix organization (green) and actin cytoskeleton (pink). (G) Based on STRING DB analysis, cluster B contained functional sub-networks of deubiquitination (yellow), laminin interaction (red), Rho GTPase effector (green), and ECM receptor interaction (blue). [Figure 11-2] See description of Figure 11-1. [Figure 11-3] See description of Figure 11-1. [Figure 11-4] See description of Figure 11-1. [Figure 12A] FIG. 12: Inhibition of myofibroblast transdifferentiation and contractility in a SMURF2-dependent manner. (A) Confocal microscopy images of phLFs treated simultaneously with TGFβ1 and active N23P and stained with αSMA (red) and Hoechst (blue) with depletion of αSMA-positive myofibroblasts. Scale = 500 μm. (B) Quantification of myofibroblasts by mean fluorescence intensity (MFI) shown in (A) showing significant inhibition of myofibroblast transdifferentiation. n = 3 (phLFs of three different patients). (C) 3D collagen gel contractility assay (scale = 2000 μm) and its quantification demonstrating significant inhibition of cell contractility by (D) Example 84. n = 3 (phLFs of three different patients). (E) Depletion of SMURF2 in IPF-phLFs by siRNA knockdown showing significant reduction of gene expression by >70%. n = 3 (phLFs of three different patients). (F) Protein expression analysis by Western blotting showing significant upregulation of αSMA protein expression in SMURF2-depleted IPF-phLF+TGFβ1+Example 84. n=4 (phLFs of 4 different patients). (G) Confocal microscopy images of SMURF2-depleted IPF-phLF+TGFβ1+Example 84 stained with αSMA (red) and Hoechst (blue). Scale=200 μm. (H) Diagram showing possible mechanism of action of N23P by SMURF2 inhibiting TGFβ1 signaling and preventing fibroblast-myofibroblast transdifferentiation. ns=not significant. *p<0.05, **p<0.01 and ***p<0.001. Statistics: one-way ANOVA with Bonferroni correction. All quantitative data represent mean±SEM. [Figure 12B] See legend to Figure 12A. [Figure 12C] See legend to Figure 12A. [Figure 12D] See legend to Figure 12A. [Figure 12E] See legend to Figure 12A. [Figure 12F] See legend to Figure 12A. [Figure 12G] See legend to Figure 12A. [Figure 12H] See legend to Figure 12A. [Figure 13A] Figure 13: Human ex vivo fibrosis model of precision-cut lung slices (PCLS) supports the anti-fibrotic effect of N23P. (A) Human ex vivo fibrosis model derived from human lung resection. (B) Volcano plot showing significantly (p < 0.05) differentially expressed proteins (pink) in fibrosis cocktail (FC)-treated PCLS and their inhibition by Example 84 and Tranilast. The top 10 most and least abundant proteins are highlighted in red and blue, respectively. (C) Heatmap showing the abundance of proteins listed in (B) and demonstrating protein deregulation as a result of Example 84 and Tranilast treatment. (D) UMAP-RPC clustering of 580 differentially expressed proteins (p < 0.1) according to similarity in expression patterns of the various conditions tested, i.e., fibrosis cocktail (FC), control cocktail (CC), FC + Example 84, and FC + Tranilast (TL). Colored boxes indicate clusters of proteins showing common abundance between the different conditions tested. (E) Analysis of commonly regulated protein abundance clusters seen in (D). (F) Immunoplots and quantification showing reduction of secreted profibrotic PAI-1 / SERPINE1 by Example 84 in live human PCLS. (G) ELISA data showing reduction of profibrotic CXCL8 / IL-8 by Example 84 in live human PCLS. * p<0.05 and ** p<0.01. Statistics: One-way ANOVA with Bonferroni correction. All quantitative data represent mean ± SEM. n = 3 (PCLS from 3 different patients). [Figure 13B] See legend to Figure 13A. [Figure 13C] See legend to Figure 13A. [Figure 13D] See legend to Figure 13A. [Figure 13E] See legend to Figure 13A. [Figure 13F]See legend to Figure 13A. [Figure 13G] See legend to Figure 13A. [Figure 14A]Figure 14: Dose-response relationship of N23P, live imaging of ECM deposition, and cell morphology changes. (A) Structural formulas and IC50 values ​​of tranilast and N23P (Example 84 and Example 85) showing 2-butoxy substitution at R1. (B) Dose-response curves of collagen V and fibrin 1 ECM deposition of N23P (Example 84 and Example 85) compared to tranilast to determine IC50 values. n = 3 (phLF of three different patients. (C) Table of IC50 of active N23P compared to tranilast. (C) 3D confocal images shown as maximum intensity projection images of deposited ECM acquired as frames from live cell experiments demonstrating dynamic ECM deposition in phLF, phLF+TGFβ1, and phLF+TGFβ1+Example 84 at 48 h. Immunostained collagen I and fibrin 1 shown as one common signal (=ECM deposition) as gold look-up table (LUT). Cell nuclei are in blue (Hoechst). Scale = 500 μm. (D) Quantification by mean fluorescence intensity (MFI) of dynamic deposition of collagen I and fibrin 1 in phLF, phLF+TGFβ1, and phLF+TGFβ1+Example 84, highlighting inhibition of ECM deposition over time by Example 84. (E) Confocal images of calcein stained confluent phLF+TGFβ1 demonstrating viability and identifying morphological changes from elongated to round cells in untreated and Example 84 treated cells. Cell nuclei were stained with Hoechst (blue). Scale = 50 μm. (F) Confocal images of subconfluent phLF+TGFβ1 stained with Hoechst (blue), calcein (green), and phalloidin (red) showing round cell morphology and extensive actin cytoskeleton reorganization (red) after treatment with Example 84. Software-based segmentation of cells within the images by CellProfiler allowed statistical analysis showing significant changes in cell shape and eccentricity to round cells after treatment with Example 84. Scale = 500 μm. Data are presented as mean ± SEM. Differences between groups were evaluated by paired t-test. * p<0.05. n = 3 (phLF of 3 different patients). [Figure 14B] See legend to Figure 14A. [Figure 14C]See legend to Figure 14A. [Figure 14D] See legend to Figure 14A. [Figure 14E] See legend to Figure 14A. [Figure 14F] See legend to Figure 14A. [Figure 15D] Figure 15: (D) Dose response curves of collagen V and fibulin 1 ECM deposition for N23P (Example 86 and Example 87) compared to tranilast to determine IC50 values. n = 3 (phLF of 3 different patients). (E) Dose response curves of collagen V and fibulin 1 ECM deposition for N23P (Example 61 and Example 88) compared to tranilast to determine IC50 values. n = 3 (phLF of 3 different patients). (F) Human dermal fibroblasts (n = 3) treated with N23P at a working concentration of 50 nM demonstrating reduced ECM deposition. Scale = 500 μm. Fibulin 1 is stained red and cell nuclei are stained with Hoechst (blue). (G) Quantification of 3D fibulin 1 deposition shown in Figure 15D normalized to cell number. ns = not significant. ** p < 0.01 and *** p < 0.001. Statistics: One-way ANOVA with Bonferroni correction. All quantitative data represent the mean ± SEM. n = 3 (human skin fibroblasts from 3 different individuals). (H) Overlay of the chemical structures of active N23P discovered by "catalog" SAR, all characterized by 2-butoxy substitution at R1. (I) Overlay of the chemical structures of active and custom-synthesized N23P with 2-butoxy or 2-o-benzyl substitution at R1. [Figure 15E] See legend to Figure 15D. [Figure 15F] See legend to Figure 15D. [Figure 15G] See legend to Figure 15D. [Figure 15H] See legend to Figure 15D. [Figure 15I] See legend to Figure 15D. [Figure 16A]FIG. 16: (A) Fluorescent wide-field microscopy images (10x objective at low magnification and resolution) of phLFs treated with 5 μM DMSO, 50 μM Example 84, and 100 μM Tranilast and stained with Hoechst (blue), α-tubulin (green), and filamentous actin (red, phalloidin). As already noted in FIG. 14(F), cells treated with Example 84 showed a cell shape conversion to a round cell morphology. phLFs treated with Tranilast or DMSO alone showed elongated cell morphology, not round, with filamentous actin (red) and microtubule filaments (green). Importantly, phLFs treated with Example 84 showed depolymerization, i.e., non-filamentous microtubules (green), without affecting actin filaments. Scale = 100 μm. (B) Fluorescent widefield microscopy images (63x objective with high magnification and resolution) of phLFs treated with 5 μM DMSO or 50 μM Example 84 and stained with Hoechst (blue), α-tubulin (green), and filamentous actin (red, phalloidin). These images clearly show intact microtubule filaments (green) in DMSO-treated phLFs, whereas Example 84-treated phLFs show mostly depolymerized (nonfilamentous) α-tubulin staining (green dots). Scale = 10 μm. In conclusion, this means that N23P (here Example 84) interferes with the regulation of microtubule dynamics and / or polymerization, whereas phLFs treated with Tranilast showed no effect on the microtubule cytoskeleton. This further speaks to the unique mechanism of action of N23P. (C) Brightfield microscopy of human lung organoids derived from human lung progenitor cells, showing the growth of organoids after 14 days of culture, either when cultured in cell culture medium alone or with DMSO. However, treatment of human organoids with 10 μM of Example 84 inhibited the growth of organoids. Quantification was performed by counting the amount of organoids found in one well divided by the amount of human lung progenitor cells seeded, and colony formation efficiency was expressed as %. Scale = 700 μm. iPSC = induced pluripotent stem cells.(D) Confocal fluorescence microscopy images of calcein (green represents live cells or organoids) and ethidium homodimer III (red represents dead cells or organoids) demonstrating that many lung progenitor cells are still viable after treatment with 10 μM of Example 84 and 14 days of culture. Scale = 350 μm. In conclusion, N23P (here Example 84) inhibits the growth of human lung organoids derived from human lung progenitor cells or stem cells, implying inhibition of stem cell differentiation and / or proliferation. Thus, the compounds of the present invention show a similar colchicine-like effect on microtubules (41). Thus, the compounds of the present invention are also used for use in the treatment of inflammatory diseases, particularly arthrolithiasis, familial Mediterranean fever and pericarditis, by inhibiting polymorphonuclear leukocyte and macrophage motility and intracellular vesicle transport mechanisms. [Figure 16B] See legend to Figure 16A. [Figure 16C] See legend to Figure 16A. [Figure 16D] See legend to Figure 16A. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] Detailed Description of the Invention The solutions of the present invention are described below, exemplified in the accompanying embodiments, shown in the drawings and reflected in the claims.

[0016] definition The term "alkyl" refers to a monoradical of a saturated straight chain or branched hydrocarbon. Preferably, the alkyl group contains 1 to 10 carbon atoms, i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms), more preferably 1 to 8 carbon atoms, e.g., 1 to 6 or 1 to 4 carbon atoms. Exemplary alkyl groups include methyl, ethyl, propyl, iso-propyl, butyl, iso-butyl, tert-butyl, n-pentyl, iso-pentyl, sec-pentyl, neo-pentyl, 1,2-dimethyl-propyl, isoamyl, n-hexyl, isohexyl, sec-hexyl, n-heptyl, isoheptyl, n-octyl, 2-ethyl-hexyl, n-nonyl, n-decyl, and the like.

[0017] The term "cycloalkyl" refers to the cyclic, non-aromatic forms of "alkyl" and "alkenyl," preferably having 3 to 10 carbon atoms, i.e., 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, more preferably 3 to 8 carbon atoms, and even more preferably 3 to 7 carbon atoms. Exemplary cycloalkyl groups include cyclopropyl, cyclopropenyl, cyclobutyl, cyclobutenyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclooctenyl, cyclononyl, cyclononenyl, and cyclodecyl. Preferred examples of cycloalkyl include (C3-C 8) -Cycloalkyl, including in particular cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl.

[0018] In the alkyl or cycloalkyl group, one or more hydrogen atoms may be substituted by halogen atoms such as Cl, Br, F, preferably F.

[0019] The term "alkenyl" refers to a monoradical of an unsaturated straight-chain or branched hydrocarbon having at least one carbon-carbon double bond. In general, the maximum number of carbon-carbon double bonds in an alkenyl group can be equal to an integer calculated by dividing the number of carbon atoms in the alkenyl group by 2, and rounding down the result of the division to the next integer if the number of carbon atoms in the alkenyl group is not an even number. For example, for an alkenyl group having 9 carbon atoms, the maximum number of carbon-carbon double bonds is 4. Preferably, the alkenyl group has 1 to 4, i.e., 1, 2, 3, or 4 carbon-carbon double bonds. Preferably, the alkenyl group contains 2 to 10 carbon atoms, i.e., 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, more preferably 2 to 8 carbon atoms, e.g., 2 to 6 carbon atoms or 2 to 4 carbon atoms. Thus, in preferred embodiments, an alkenyl group contains from 2 to 10 carbon atoms and 1, 2, 3, 4, or 5 carbon-carbon double bonds, more preferably from 2 to 8 carbon atoms and 1, 2, 3, or 4 carbon-carbon double bonds, e.g., from 2 to 6 carbon atoms and 1, 2, or 3 carbon-carbon double bonds, or from 2 to 4 carbon atoms and 1 or 2 carbon-carbon double bonds. The carbon-carbon double bonds may be in the cis (Z) or trans (E) configuration. Exemplary alkenyl groups are vinyl, 1-propenyl, 2-propenyl (i.e., allyl), 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 5-heptenyl, 6-heptenyl, Examples of alkenyl groups include 1-octenyl, 1-octenyl, 2-octenyl, 3-octenyl, 4-octenyl, 5-octenyl, 6-octenyl, 7-octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, 4-nonenyl, 5-nonenyl, 6-nonenyl, 7-nonenyl, 8-nonenyl, 1-decenyl, 2-decenyl, 3-decenyl, 4-decenyl, 5-decenyl, 6-decenyl, 7-decenyl, 8-decenyl, 9-decenyl, etc. If the alkenyl group is attached to a nitrogen atom, the double bond cannot be alpha to the nitrogen atom.

[0020] The term "alkenylene" refers to an unsaturated linear or branched hydrocarbon diradical having at least one carbon-carbon double bond. In general, the maximum number of carbon-carbon double bonds in an alkenylene group can be equal to an integer calculated by dividing the number of carbon atoms in the alkenylene group by 2, and rounding down the result of the division to the next integer if the number of carbon atoms in the alkenylene group is not an even number. For example, for an alkenylene group having 9 carbon atoms, the maximum number of carbon-carbon double bonds is 4. Preferably, the alkenylene group has 1 to 4, i.e., 1, 2, 3, or 4 carbon-carbon double bonds. Preferably, the alkenylene group contains 2 to 10 carbon atoms, i.e., 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, more preferably 2 to 8 carbon atoms, e.g., 2 to 6 carbon atoms or 2 to 4 carbon atoms. Thus, in preferred embodiments, the alkenylene group contains 2 to 10 carbon atoms and 1, 2, 3, 4, or 5 carbon-carbon double bonds, more preferably 2 to 8 carbon atoms and 1, 2, 3, or 4 carbon-carbon double bonds, e.g., 2 to 6 carbon atoms and 1, 2, or 3 carbon-carbon double bonds, or 2 to 4 carbon atoms and 1 or 2 carbon-carbon double bonds. The carbon-carbon double bonds may be in the cis (Z) or trans (E) configuration. Exemplary alkenylene groups include ethene-1,2-diyl, vinylidene, 1-propene-1,2-diyl, 1-propene-1,3-diyl, 1-propene-2,3-diyl, allylidene, 1-butene-1,2-diyl, 1-butene-1,3-diyl, 1-butene-1,4-diyl, 1-butene-2,3-diyl, 1-butene-2,4-diyl, 1-butene-3,4-diyl, 2-butene-1,2-diyl, 2-butene-1,3-diyl, 2-butene-1,4-diyl, 2-butene-2,3-diyl, 2-butene-2,4-diyl, 2-butene-3,4-diyl, and the like. When an alkenylene group is attached to a nitrogen atom, the double bond cannot be alpha to the nitrogen atom.

[0021] The term "alkynyl" refers to an unsaturated linear or branched hydrocarbon monoradical having at least one carbon-carbon triple bond. In general, the maximum number of carbon-carbon triple bonds in an alkynyl group can be equal to an integer calculated by dividing the number of carbon atoms in the alkynyl group by 2, and rounding down the result of the division to the next integer if the number of carbon atoms in the alkynyl group is not an even number. For example, for an alkynyl group having 9 carbon atoms, the maximum number of carbon-carbon triple bonds is 4. Preferably, the alkynyl group has 1 to 4 carbon-carbon triple bonds, i.e., 1, 2, 3, or 4, more preferably 1 or 2 carbon-carbon triple bonds. Preferably, the alkynyl group contains 2 to 10 carbon atoms, i.e., 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, more preferably 2 to 8 carbon atoms, e.g., 2 to 6 carbon atoms or 2 to 4 carbon atoms. Thus, in preferred embodiments, an alkynyl group contains 2 to 10 carbon atoms and 1, 2, 3, 4, or 5 (preferably 1, 2, or 3) carbon-carbon triple bonds, more preferably 2 to 8 carbon atoms and 1, 2, 3, or 4 (preferably 1 or 2) carbon-carbon triple bonds, for example, 2 to 6 carbon atoms and 1, 2, or 3 carbon-carbon triple bonds, or 2 to 4 carbon atoms and 1 or 2 carbon-carbon triple bonds. Exemplary alkynyl groups are ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, 5-hexynyl, 1-heptynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 5-heptynyl, 6-heptynyl, 1-octynyl. , 2-octynyl, 3-octynyl, 4-octynyl, 5-octynyl, 6-octynyl, 7-octynyl, 1-nonylyl, 2-nonynyl, 3-nonynyl, 4-nonynyl, 5-nonynyl, 6-nonynyl, 7-nonynyl, 8-nonynyl, 1-decynyl, 2-decynyl, 3-decynyl, 4-decynyl, 5-decynyl, 6-decynyl, 7-decynyl, 8-decynyl, 9-decynyl, etc. If the alkynyl group is attached to a nitrogen atom, the triple bond cannot be alpha to the nitrogen atom.

[0022] The term "heterocyclyl" means a cycloalkyl group, as defined above, in which 1, 2, 3, or 4 carbon atoms in the cycloalkyl group are replaced with an O, S, or N heteroatom. Preferably, in each ring of a heterocyclyl group, the maximum number of O atoms is 1, the maximum number of S atoms is 1, and the maximum total number of O and S atoms is 2. The term "heterocyclyl" is also meant to include partially or fully hydrogenated forms (e.g., dihydro, tetrahydro or perhydro forms) of the above heteroaryl groups. Exemplary heterocyclyl groups are morpholino, isochromanyl, chromanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperidinyl, piperazinyl, indolinyl, isoindolinyl, di- and tetrahydrofuranyl, di- and tetrahydrothienyl, di- and tetrahydrooxazolyl, di- and tetrahydroisoxazolyl, di- and tetrahydrooxadiazolyl (1,2,5- and 1,2,3-), dihydropyrrolyl, dihydroimidazole, dihydroisoxazolyl ... Dazolyl, dihydropyrazolyl, di- and tetrahydrotriazolyl (1,2,3- and 1,2,4-), di- and tetrahydrothiazolyl, di- and tetrahydrothiazolyl, di- and tetrahydrothiadiazolyl (1,2,3- and 1,2,5-), di- and tetrahydropyridyl, di- and tetrahydropyrimidinyl, di- and tetrahydropyrazinyl, di- and tetrahydrotriazinyl (1,2,3-, 1,2,4-, and 1,3,5-), di- and tetrahydrobenzofuranyl (1- and 2-), di- and tetrahydroindolyl, di- and tetrahydroisoindolyl, di- and tetrahydrobenzothienyl (1- and 2), di- and tetrahydro-1H-indazolyl, di- and tetrahydrobenzimidazolyl, di- and tetrahydrobenzoxazolyl, di- and tetrahydroindoxazinyl, di- and tetrahydrobenzisoxazolyl, di- and tetrahydrobenzothiazolyl, di- and tetrahydro dihydrobenzoisothiazolyl, di- and tetrahydrobenzotriazolyl, di- and tetrahydroquinolinyl, di- and tetrahydroisoquinolinyl, di- and tetrahydrobenzodiazinyl, di- and tetrahydroquinoxalinyl, di- and tetrahydroquinazolinyl, di- and tetrahydrobenzotriazinyl (1,2,3- and 1,2,4-), di- and tetrahydropyridazinyl, di- and tetrahydrophenoxazinyl, di- and tetrahydrothiazolopyridinyl (e.g. 4,5,6 -7-tetrahydro[1,3]thiazolo[5,4-c]pyridinyl or 4,5,6-7-tetrahydro[1,3]thiazolo[4,5-c]pyridinyl, for example 4,5,6-7-tetrahydro[1,3]thiazolo[5,4-c]pyridin-2-yl or 4,5,6-7-tetrahydro[1,3]thiazolo[4,5-c]pyridin-2-yl), di- and tetrahydropyrrolothiazolyl (for example 5,6-dihydro-4H-pyrrolo[3,4-d][1,3]thiazolyl), di- and tetrahydrophenothiazinyl di- and tetrahydroisobenzofuranyl, di- and tetrahydrochromenyl, di- and tetrahydroxanthenyl, di- and tetrahydrophenoxathiinyl, di- and tetrahydropyrrolidinyl, di- and tetrahydroindolizinyl, di- and tetrahydroindazolyl, di- and tetrahydropurinyl, di- and tetrahydroquinolizinyl, di- and tetrahydrophthalazinyl, di- and tetrahydronaphthyridinyl (1,5-, 1,6-, 1,7-, 1,8-, and 2,di- and tetrahydrocinnolinyl, di- and tetrahydropteridinyl, di- and tetrahydrocarbazolyl, di- and tetrahydrophenanthridinyl, di- and tetrahydroacridinyl, di- and tetrahydroperimidinyl, di- and tetrahydrophenanthrolinyl (1,7-, 1,8-, 1,10-, 3,8-, and 4,7-), di- and tetrahydrophenazinyl, di- and tetrahydrooxazolopyridinyl, di- and tetrahydroisoxazolopyridinyl, di- and tetrahydropyrrolooxazolyl, and di- and tetrahydropyrrolopyrrolyl. Exemplary 5- or 6-membered heterocyclyl groups are morpholino, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperidinyl, piperazinyl, di- and tetrahydrofuranyl, di- and tetrahydrothienyl, di- and tetrahydrooxazolyl, di- and tetrahydroisoxazolyl, di- and tetrahydrooxadiazolyl (1,2,5- and 1,2,3-), dihydropyrrolyl, dihydroimidazolyl, dihydropyrazolyl, di- and tetrahydrothionyl, These include thiazolyl (1,2,3- and 1,2,4-), di- and tetrahydrothiazolyl, di- and tetrahydroisothiazolyl, di- and tetrahydrothiadiazolyl (1,2,3- and 1,2,5-), di- and tetrahydropyridyl, di- and tetrahydropyrimidinyl, di- and tetrahydropyrazinyl, di- and tetrahydrotriazinyl (1,2,3-, 1,2,4-, and 1,3,5-), and di- and tetrahydropyridazinyl.

[0023] The term "aryl" refers to a monoradical of an aromatic cyclic hydrocarbon. Preferably, an aryl group contains 3 to 10 (e.g., 5 to 10, e.g., 5, 6, or 10) carbon atoms which may be arranged in one ring (e.g., phenyl) or two or more fused rings (e.g., naphthyl). Exemplary aryl groups include cyclopropenylium, cyclopentadienyl, phenyl, indenyl, naphthyl, azulenyl, fluorenyl, anthryl, and phenanthryl. Preferably, "aryl" refers to a monocyclic ring containing 6 carbon atoms or an aromatic bicyclic ring system containing 10 carbon atoms. Preferred examples are phenyl and naphthyl.

[0024] The term "heteroaryl" refers to an aryl group as defined above, in which one or more carbon atoms in the aryl group are replaced by a heteroatom of O, S, or N. Preferably, heteroaryl refers to a 5- or 6-membered aromatic monocyclic ring in which 1, 2, or 3 carbon atoms are replaced by the same or different heteroatoms of O, N, or S. Alternatively, it refers to an aromatic bicyclic or tricyclic ring system in which 1, 2, 3, 4, or 5 carbon atoms are replaced by the same or different heteroatoms of O, N, or S. Preferably, in each ring of the heteroaryl group, the maximum number of O atoms is 1, the maximum number of S atoms is 1, and the maximum total number of O and S atoms is 2. Exemplary heteroaryl groups are furanyl, thienyl, oxazolyl, isoxazolyl, oxadiazolyl (1,2,5- and 1,2,3-), pyrrolyl, imidazolyl, pyrazolyl, triazolyl (1,2,3- and 1,2,4-), tetrazolyl, thiazolyl, isothiazolyl, thiadiazolyl (1,2,3- and 1,2,5-), pyridyl, pyrimidinyl, pyrazinyl, triazinyl (1,2,3-, 1,2,4-, and 1,3,5-), benzofuranyl (1- and 2-), indolyl, isoindolyl, benzothienyl (1- and 2-), 1H-indazolyl, benzimidazolyl, benzoxazolyl, indoxazinyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzotriazolyl, quinolinyl, isoquinolinyl, benzodiaryl, and the like. Azinyl, quinoxalinyl, quinazolinyl, benzotriazinyl (1,2,3- and 1,2,4-benzotriazinyl), pyridazinyl, phenoxazinyl, thiazolopyridinyl, pyrrolothiazolyl, phenothiazinyl, isobenzofuranyl, chromenyl, xanthenyl, phenoxathiinyl, pyrrolidinyl, indolizinyl, indazolyl, purinyl, quinolizinyl, phthalazinyl, naphthalenyl, These include phthyridinyl (1,5-, 1,6-, 1,7-, 1,8-, and 2,6-), cinnolinyl, pteridinyl, carbazolyl, phenanthridinyl, acridinyl, perimidinyl, phenanthrolinyl (1,7-, 1,8-, 1,10-, 3,8-, and 4,7-), phenazinyl, oxazolopyridinyl, isoxazolopyridinyl, pyrrolooxazolyl, and pyrrolopyrrolyl.Exemplary 5- or 6-membered heteroaryl groups include furanyl, thienyl, oxazolyl, isoxazolyl, oxadiazolyl (1,2,5- and 1,2,3-), pyrrolyl, imidazolyl, pyrazolyl, triazolyl (1,2,3- and 1,2,4-), thiazolyl, isothiazolyl, thiadiazolyl (1,2,3- and 1,2,5-), pyridyl, pyrimidinyl, pyrazinyl, triazinyl (1,2,3-, 1,2,4-, and 1,3,5-), and pyridazinyl.

[0025] The term "azido" refers to N3.

[0026] In an alkyl, cycloalkyl, heterocyclyl, alkenyl, alkenylene, aryl, or heteroaryl group, one or more hydrogen atoms may be replaced by a halogen atom such as Cl, Br, F, preferably F, -OH, -NH2, -NHC(O)CH3, -CN, -N3, -COOH, and / or -C(O)NH.

[0027] The present invention includes compounds according to formula (I): TIFF2024527473000011.tif42128 expression R 1 -OR 12 , -O(CH2) u (C3~C 10 )Aryl, -O(CH2) u (C2) Alkynyl; -(CH2) u (C3~C 10 )Aryl, -O(CH2) u (C3~C 10 ) cycloalkyl, -(CH2) u (C3~C 10 ) cycloalkyl, TIFF2024527473000012.tif17128, -(CH2) u (C2) alkynyl.

[0028] Preferably, For TIFF2024527473000013.tif17128, u is 1 and (C1-C6) alkyl is methyl.

[0029] u is 0 to 6.

[0030] R 2 ~R 5 are independently H, -OR 12 , -(C1~C 10 )Alkyl, halogen, cyano, isocyano, cyanato, isocyanato, thiocyanato, isothiocyanato, azido, -(C2-C 10 ) alkenyl, -(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 )Heterocyclyl, -(C3~C 10 )Aryl, -(C3~C 10 )heteroaryl, -CHZ2, -CZ3-CH2Z, -OCHZ2, -OCZ3, -OCH2Z- -N(R 13 )(R 14 ), -N(R 15 )(OR 16 ), -S(O) 0~2 R 17 , -S(O) 1~2 OR 18 , -OS(O) 1~2 R 19 , -OS(O) 1~2 OR 20 , -S(O) 1~2 N(R 21 )(R 22 ), -OS(O) 1~2 N(R 23 )(R 24 ), -N(R 25 )S(O) 1~2 R 26 , -NR 27 S(O) 1~2 OR 28 , -NR 29 S(O) 1~2 N(R 30 )(R 31 ), -C(=X)R 32 , -C(=X)XR 33 , -XC(=X)R34 , and -XC(=X)XR 35 , -OR 36 , -O(CH2) v (C3~C 10 )Aryl, -O(CH2) v (C3~C 10 ) Cycloalkyl, -O(CH2) v (C2) alkynyl.

[0031] R 6 H, -(C1~C 10 ) alkyl, and -(CH2) 1~5 (C3~C 10 )cycloalkyl; where -(C1-C 10 ) alkyl, benzyl and -(CH2) 1~5 (C3~C 10 ) Cycloalkyl may be further substituted with at least one substituent selected from the group consisting of halogen, preferably F.

[0032] R 7 ~R 11 are independently H, -OR 12 , -SR 12 , -(C1~C 10 )Alkyl, halogen, -(C1-C 10 )AlkylO(C1-C 10 )Alkyl, cyano, isocyano, cyanato, isocyanato, thiocyanato, isothiocyanato, azido, -(C2-C 10 ) alkenyl, -(C2-C 10 ) alkynyl, -O(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 )Heterocyclyl, -(C3~C 10 )Aryl, -(C3~C 10 ) Heteroaryl, -(CH2) v CHZ2, -CZ3-CH2Z, -OCHZ2, -OCZ3, -OCH2Z, -N(R 13 )(R 14 ), -N(R 15 )(OR16 ), -S(O) 0~2 R 17 , -S(O) 1~2 OR 18 , -OS(O) 1~2 R 19 , -OS(O) 1~2 OR 20 , -S(O) 1~2 N(R 21 )(R 22 ), -OS(O) 1~2 N(R 23 )(R 24 ), -N(R 25 )S(O) 1~2 R 26 , -NR 27 S(O) 1~2 OR 28 , -NR 29 S(O) 1~2 N(R 30 )(R 31 ), -C(=X)R 32 , -C(=X)XR 33 , -XC(=X)R 34 , and -XC(=X)XR 35 , -O(CH2) v (C3~C 10 ) Cycloalkyl, -O(CH2) v (C1~C 10 ) alkyl and -O(CH2) v (C3~C 10 ) aryl.

[0033] R 1 ~R 5 and R 7 ~R 11 , preferably R 8 ~R 10 The two adjacent remaining moieties of the formula (III) to (XI): Optionally, a ring may be formed attached to the basic aromatic ring of formula (I) according to TIFF2024527473000014.tif25162; In the formula, T 1 and T 2 are independently H, -(C1 to C 10 ) selected from the group consisting of alkyl and halogen; Here, each hydrogen in the formulas (III) to (XI) is a halogen or -(C 10 ) aryl, preferably F. R 38 are independently H, -(C1 to C 10 ) alkyl; Het is selected from O, S, NH. G is selected from CH and NH.

[0034] J1 to J4 are independently selected from C or N, and preferably J1 to J4 are C; where, when any one of J1 to J4 is N, the corresponding R bonded to each of J1 to J4 which is N is 1 ~R 4 does not exist.

[0035] R 12 ~R 36 are independently H, -(C1 to C 10 )Alkyl, -(C2-C 10 ) alkenyl, -(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 )Heterocyclyl, -(C3~C 10 )Aryl, -(C3-C 10 ) heteroaryl.

[0036] Independently - (C1~C 10 )Alkyl, -(C2-C 10 ) alkenyl, -(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 )Heterocyclyl, -(C3~C 10 )Aryl, -O(CH2) v (C3~C 10 ) Cycloalkyl, -O(CH2) v (C1~C 10 ) alkyl and -O(CH2) v (C3~C 10 ) R selected from the group consisting of aryl1 ~R 11 , and R 12 ~R 35 OR 12 , -(C1~C 10 )Alkyl, halogen, cyano, isocyano, cyanato, isocyanato, thiocyanato, isothiocyanato, azido, -(C2-C 10 ) alkenyl, -(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 )Heterocyclyl, -(C3~C 10 )Aryl, -(C3~C 10 )heteroaryl, -CHZ2, -CZ3-CH2Z, -OCHZ2, -OCZ3, -OCH2Z-, -N(R 13 )(R 14 ), -N(R 15 )(OR 16 ), -NHC(O)(C1-C 10 ) alkyl, -S(O) 0~2 R 17 , -S(O) 1~2 OR 18 , -OS(O) 1~2 R 19 , -OS(O) 1~2 OR 20 , -S(O) 1~2 N(R 21 )(R 22 ), -OS(O) 1~2 N(R 23 )(R 24 ), -N(R 25 )S(O) 1~2 R 26 , -NR 27 S(O) 1~2 OR 28 , -NR 29 S(O) 1~2 N(R 30 )(R 31 ), -C(=X)R 32 , -C(=X)XR 33 , -XC(=X)R 34 , and -XC(=X)XR 35 , -OR 36 , and -O(CH2) v (C3~C10 ) may be further substituted with at least one substituent selected from the group consisting of aryl.

[0037] v is 0 to 5.

[0038] Z is a halogen.

[0039] X is selected from the group consisting of O, -NH- or S.

[0040] A is TIFF2024527473000015.tif36128; n is 1, 2, or 3, preferably 1; o is 1, 2, or 3, preferably 1; R is H, (C1-C6) alkyl, cyano, -(C3-C 10 ) cycloalkyl, benzyl, or R is TIFF2024527473000016.tif15128 by R 7 or R 11 is part of a ring connecting However, if n is 2 or 3, A is Could be TIFF2024527473000017.tif15128; R 37 is H or -CF3.

[0041] However, R 5 is not -COOH.

[0042] In one embodiment, I)R 1 -OR 12 , -O(CH2) u (C3~C 10 )Aryl, -O(CH2) u (C3~C 10 ) Cycloalkyl, -O(CH2) u (C2) the group consisting of alkynyl; preferably -OR12 More selected; u is 0 to 5; R 12 Ha-(C1~C 10 ) alkyl, preferably -(C3-C5) alkyl, more preferably -(C4) alkyl, and / or II) A is Selected from TIFF2024527473000018.tif9128.

[0043] In another embodiment, R 2 ~R 5 are independently H, -OR 12 , -(C1~C 10 )Alkyl, halogen, cyano, isocyano, cyanato, isocyanato, thiocyanato, isothiocyanato, azido, -(C2-C 10 ) alkenyl, -(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 )Heterocyclyl, -(C3~C 10 )Aryl, -(C3~C 10 )heteroaryl, -CHZ2, -CZ3-CH2Z, -OCHZ2, -OCZ3, -OCH2Z, -N(R 13 )(R 14 ), -N(R 15 )(OR 16 ), -C(=X)R 32 , -C(=X)XR 33 , -XC(=X)R 34 , and -XC(=X)XR 35 , -O(CH2) v (C3~C 10 )Aryl, -O(CH2) v (C3~C 10 ) Cycloalkyl, -O(CH2) v (C2) alkynyl.

[0044] In another embodiment, R 7 ~R 11 are independently H, -SR 12 , -OR 12 , -(C1~C10 )Alkyl, halogen, cyano, isocyano, cyanato, isocyanato, thiocyanato, isothiocyanato, azido, -(C2-C 10 ) alkenyl, -(C2-C 10 ) alkynyl, -O(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 )Heterocyclyl, -(C3~C 10 )Aryl, -(C3-C 10 ) Heteroaryl, -(CH2) v CHZ2, -CZ3-CH2Z, -OCHZ2, -OCZ3, -OCH2Z, -N(R 13 )(R 14 ), -N(R 15 )(OR 16 ), -C(=X)R 32 , -C(=X)XR 33 , -XC(=X)R 34 , and -XC(=X)XR 35 , -O(CH2) v (C3~C 10 ) Cycloalkyl, -O(CH2) v (C1~C 10 ) alkyl and -O(CH2) v (C3~C 10 ) aryl.

[0045] Here, R 1 ~R 5 and R 7 ~R 11 The two adjacent remaining parts of Optionally, a ring may be formed attached to the basic aromatic ring of formula (II) according to TIFF2024527473000019.tif25162; In the formula, T 1 and T 2 are independently H, -(C1 to C 10 ) selected from the group consisting of alkyl and halogen; Here, each hydrogen in the formulas (III) to (XI) is a halogen or -(C 10 ) aryl, preferably F. Het is selected from O, S, NH. G is selected from CH and NH. R 38 are independently H, -(C1 to C 10 ) alkyl.

[0046] In a further embodiment, -(C 10 )Alkyl, -(C2-C 10 ) alkenyl, -(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 )Heterocyclyl, -(C3~C 10 )Aryl, -O(CH2) v (C3~C 10 ) Cycloalkyl, -O(CH2) v (C1~C 10 ) alkyl and -O(CH2) v (C3~C 10 ) R selected from the group consisting of aryl 1 ~R 11 , and R 12 ~R 35 -OR 12 , -(C1~C 10 )Alkyl, halogen, cyano, isocyano, cyanato, isocyanato, thiocyanato, isothiocyanato, azido, -(C2-C 10 ) alkenyl, -(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 )Heterocyclyl, -(C3~C 10 )Aryl, -(C3-C 10 )heteroaryl, -CHZ2, -CZ3-CH2Z, -OCHZ2, -OCZ3, -OCH2Z, -N(R 13 )(R 14 ), -N(R 15 )(OR 16 ), -C(=X)R 32 , -C(=X)XR 33 , -XC(=X)R 34 , and -XC(=X)XR 35 , -OR36 , and -O(CH2) v (C3~C 10 R may be further substituted with a substituent selected from the group consisting of aryl. 12 -OR 12 When substituted with further substituents such as 12 is basic R 12 and its substituent -OR 12 may be different in

[0047] In a further aspect, R 9 -OR 12 , halogen, -O(C2~C 10 ) Alkynyl, -CZ3, -OCHZ2, -OCZ3, -OCH2Z, -O(CH2) v (C3~C 10 ) Cycloalkyl, -O(CH2) v (C1~C 10 ) alkyl and -O(CH2) v (C3~C 10 )aryl, where -O(C1-C 10 )Alkyl, -OCH2Z, -O(CH2) v (C3~C 10 ) Cycloalkyl, -O(CH2) v (C1~C 10 ) alkyl and -O(CH2) v (C3~C 10 ) R selected from the group consisting of aryl 9 may be further substituted with at least one substituent selected from the group consisting of halogen, -OH, -NH2, -NHC(O)CH3, -CN, -N3, and -COOH, -C(O)NH2.

[0048] Additionally, the present invention includes compounds according to formula (II). TIFF2024527473000020.tif47128

[0049] R 1 ~R 4 and R 6 , R7 , R 9 and R 10 are independently H, -(C1 to C 10 ) alkyl, -SR 12 , halogen, azide, cyano, -O(C1-C 10 ) alkyl, -(CH2) u (C3~C 10 )Aryl, -O(CH2) u (C3~C 10 ) cycloalkyl, -(CH2) u (C3~C 10 )Cycloalkyl, -(C2-C 10 ) alkenyl, TIFF2024527473000021.tif17128-(C2~C 10 ) alkynyl.

[0050] R 5 Ha-(C1~C 10 ) alkyl, -(C5-C6) heteroaryl, -(C3-C 10 ) Aryl, -(CH2) u (C3~C 10 ) cycloalkyl, preferably -(C4-C6) alkyl, or benzyl, and most preferably -(C4) alkyl.

[0051] R 8 is H, -O(C1-C 10 ) alkyl, -O(CH2) u (C3~C 10 )Aryl, -O(CH2) u (C3~C 10 )Cycloalkyl, -O(C3-C 10 ) cycloalkyl, or -O(C2-C 10 ) alkenyl, preferably -OCH3.

[0052] u is 0 to 6.

[0053] R is H, (C1-C6) alkyl, cyano, -(C3-C 10 ) cycloalkyl, benzyl, or R is TIFF2024527473000022.tif14128 by R 7 or R 11 and is preferably H or benzyl, most preferably H.

[0054] R 11 is H, -(C1~C 10 ) alkyl, and -(CH2) 1~5 (C3~C 10 ) cycloalkyl.

[0055] Independently - (C1~C 10 ) alkyl, -O(C1-C 10 )Alkyl, -(C2-C 10 ) alkenyl, -(C2-C 10 )Alkynyl, -(C3-C 10 ) cycloalkyl, -(CH2) u (C3~C 10 )Cycloalkyl, -(C3-C 10 ) aryl, and -(CH2) u (C3~C 10 ) R selected from the group consisting of aryl 1 ~R 11 , and R 12 ~R 35 may be further substituted with at least one substituent selected from the group consisting of halogen, -OH, -NH2, -NHC(O)CH3, -CN, -N3, and -COOH, -C(O)NH2.

[0056] R 12 Ha-(C1~C 10 ) alkyl, preferably -(C3 to C5) alkyl, and more preferably -(C4) alkyl.

[0057] R 8 ~R 10 The two adjacent remaining parts of Optionally, a ring may be formed attached to the basic aromatic ring of formula (II) according to TIFF2024527473000023.tif26162; Here, R8 where the oxygen to which it is attached may correspond to one of the oxygen / Het atoms shown in formulae (III)-(XI); In the formula, T 1 and T 2 are independently H, -(C1 to C 10 ) selected from the group consisting of alkyl and halogen; Here, each hydrogen in the formulas (III) to (XI) is a halogen or -(C 10 ) aryl, preferably F.

[0058] Het is selected from O.

[0059] G is selected from CH and NH.

[0060] R 38 are independently H, -(C1 to C 10 ) alkyl.

[0061] J1 to J4 are independently selected from C or N, and preferably J1 to J4 are C.

[0062] Here, when any one of J1 to J4 is N, the corresponding R bonded to each of J1 to J4 which is N 1 ~R 4 does not exist.

[0063] In one embodiment, one of J1 to J4 is N. In another embodiment, two of J1 to J4 are N.

[0064] however, When J1 to J4 are C, and I)R 5 is -(CH2)3CH3; R 9 is -OCH3, -OCH2CH3, -O(CH2)2CH3, -OCH2phenyl or -O(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R7 , R 11 and R is H; and (a)R 8 and R 10 is H; or (b)R 8 is -OCH3 or -OCH2CH3, and R 10 is H; or (c)R 10 is -OCH3 or -OCH2CH3, and R 8 is H Case;R 6 is not H; II) R 9 is -OCH3, -O(CH2)2CH3, -O(2-propyl), -O(CH2)4CH3, -O(CH2)5CH3, -OCH2(4-chlorophenyl), -O(CH2)2CH(CH3)2, -OCH2(2,6-dichlorophenyl), or -OCH2phenyl; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R is H; R 8 is -OCH3 and R 10 is H or R 10 is -OCH3 and R 8 is H;R 5 is -(CH2)3CH3 Case;R 6 is not H; III) R 9 is -OCH3; R 8 is Br and R 10 is H or R 10 is Br and R 8 is H;R 5 is -(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 11 and R is H In this case, R 1 is not H; IV) R 5is -(CH2)3CH3, and R 9 is -OCH3, R 7 is -OCH3 and R 11 is H or R 11 is -OCH3 and R 7 is H;R 1 , R 2 , R 3 , R 4 , R 8 , R 10 , R 11 and R is H Case;R 6 is not H; V)R 9 is -OCH3, -OCH2phenyl, or -OCH2(2-fluorophenyl); R 8 is Br and R 10 is -OCH3 or R 10 is Br and R 8 is -OCH3; R 5 is -O(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R is H In this case, R 6 is not H; VI)R 9 is -O(CH2)3CH3; R 8 is -OCH2CH3 and R 10 is H or R 10 is -OCH2CH3 and R 8 is H;R 5 is -O(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R is H In this case, R 6 is not H; VII) R 9 is -OCH2 (2-chlorophenyl); R 8is Br and R 10 is -CH2CH3 or R 10 is Br and R 8 is -OCH2CH3; R 5 is -(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R is H In this case, R 6 is not H; VIII)R 9 is -O(2-octenyl); R 8 is Cl and R 10 is H or R 10 is Cl and R 8 is H;R 5 is -(CH2)3COOH; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R is H In this case, instead of H; IX)R 9 is -OCH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 8 , R 10 , R 11 , R is H; and R 5 is -(2-fluorophenyl), -phenyl Case;R 6 is not H; X)R 9 is -OCH2CH3; R 5 is -(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 8 , R 10 , R 11 and R is H In this case, R6 is not H; XI)R 9 is -OCH3; R 8 is -OCH3 and R 10 is H or R 10 is -OCH3 and R 8 is H;R 5 is -(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 8 , R 7 and R is H In this case, R 6 is not H; XII) R 9 is -OCH3; R 5 is -O(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R is H and R 8 is -OCH2CH3 and R 10 is H or R 10 is -OCH2CH3 and R 8 is H In this case, R 6 is not H; XIII)R 5 is -(CH2)3CH3; R 9 is -O(CH2)3CH3 or -OCH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 8 , R 10 , R 11 and R is H In this case, R 6 is not H; XIV)R 5 is -CH3, -(CH2)2CH3 or -CH2CH3; R 9 is -OCH3; R 8 is -OCH3 and R 10is H or R 8 is H and R 10 is -OCH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R is H Case;R 6 is not H; XV)R 5 is -(CH2)3CH3; R 1 , R 2 , R 3 , R 4 , R 7 , R 8 , R 9 , R 10 and R 11 is H Case;R 6 is not H; and / or However, when J1 to J4 are C, the following compounds are not included in the compounds according to formula (II). TIFF2024527473000024.tif91163TIFF2024527473000025.tif223163In a further embodiment, when J1 to J4 are C, the following compounds, or the following compounds, are also not included in the compounds according to formula (II): TIFF2024527473000026.tif23128

[0065] With respect to the compounds according to formula (I) or (II), the present invention further includes the following embodiments (i) to (xxiv): (i)R 9 -OR 12 and R 8 is -H and R 10 is H or -OR 12 is; or (ii)R 9 is H;R 8 is H and R 10 -OR 12 or (iii)R 9 and R10 but, forming a ring linked to the basic aromatic ring of formula (II), preferably according to formula (III), according to TIFF2024527473000027.tif24128, In the formula, T 1 and T 2 are independently H, -(C1 to C 10 ) selected from the group consisting of alkyl and halogen; Here, each hydrogen in the formulas (III) to (VII) is a halogen or -(C 10 ) optionally substituted with aryl, preferably F; R 38 are independently H, -(C1 to C 10 ) alkyl; Het is O, S or NH, preferably O; G is selected from CH, N; and / or (iv)J 1~4 is CH; and / or (v)R 2 ~R 5 , R 7 or R 11 are independent, H, -OR 12 , -(C1~C 10 )Alkyl, halogen, cyano, azide, -(C2-C 10 ) alkenyl, -(C2-C 10 )Alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 )Heterocyclyl, -(C3~C 10 )Aryl, -(C3-C 10 ) Heteroaryl, -CHZ2, -CZ3-CH2Z, -OCHZ2, -OCZ3, -OCH2Z, -OR 36 , -O(CH2) v (C3~C 10 )Aryl, -O(CH2) v (C3~C 10 ) Cycloalkyl, -O(CH2) v (C2) alkynyl; and / or (vi) u = 0 to 3; and / or (vii)R 12 is -O(C4-C6)alkyl, -OCH2(C3-C5)cycloalkyl, -Ophenyl or -OCH2phenyl; and / or (viii)R 6 = H, or C1-C4 alkyl or (CH2) (1~3) alkyl-(C1-C6)cycloalkyl; and / or (ix) R 2 ~R 5 , R 7 or R 11 are independent, H, -OR 12 , -(C1~C3) alkyl, halogen, cyano, azide, -CHZ2, -CZ3-CH2Z, -OCHZ2, -OCZ3, -OCH2Z, -O(CH2) v (C3~C 10 )Aryl, -O(CH2) v (C3~C 10 ) Cycloalkyl, -O(CH2) v (C2) alkynyl; and / or (x)R 12 is H, -(C1~C 10 ) alkyl, -(C2-C4) alkynyl, -(C3-C 10 )Cycloalkyl, -(C3-C 10 )Heterocyclyl, -(C3~C 10 )Aryl, -(C3-C 10 ) heteroaryl; and / or (xi)R 2 ~R 5 , R 7 or R 11 is independently selected from the group consisting of H, -(C1-C3)alkyl, halogen, cyano, azido, -CHZ2, -CZ3-CH2Z, -OCHZ2, -OCZ3, -OCH2Z, -(C3-C5)cycloalkyl; (xii)R 2 is H; and / or (xiii)R 7 is H; and / or (xiv)R 11 is H; and / or (xv)R 3 is H; and / or (xvi)R 4 is H; and / or (xvii)R 5 is H. (xviii)R 9 is OCH3 and R 10 is H or R 10 is OCH3, and / or (xix)R 6 is H; and / or (xx)R 1 -O(C -1 -C6)) alkyl; and / or (xxi)R 10 is H or -OCH3; and / or (xxii)R 1 is -O(C4-C6) alkyl; and / or (xxiii)R 9 is -OCH3, R 10 = H or -OCH3; and / or (xxiv)R 10 is H.

[0066] With respect to the compounds according to formula (II), the present invention further includes the following aspects: (i)R 1 -O(C1-C4) alkyl, -O(C5-C 10 ) Heteroaryl, -O(C3-C 10 )Aryl, -O(CH2) u (C3~C 10 ) aryl, preferably -O-butyl; Here, (a)(C5~C 10 Heteroaryl is preferably TIFF2024527473000028.tif19128; and / or (b) (C3~C 10 ) aryl is preferably phenyl, optionally substituted with halogen and / or (C1-C3) alkyl; and / or (ii)R 2 is selected from the group consisting of hydrogen, halogen, preferably hydrogen; wherein halogen is preferably Cl or F; (c)R 3 is selected from the group consisting of hydrogen, halogen, preferably hydrogen; wherein halogen is preferably Cl or F; and / or (iii)R 4 is selected from the group consisting of hydrogen, halogen, preferably hydrogen; wherein halogen is preferably Cl or F; and / or (iv) R 5 is selected from the group consisting of hydrogen, halogen, preferably hydrogen; wherein halogen is preferably Cl or F; and / or (v)R 6 is H, -(C1~C 10 ) alkyl, and -(CH2) 1~5 is cyclopropyl; and / or (vi)R 7 is H; and / or (vii)R 8 is H; and / or (viii)R 9 is H, -O(C1-C4) alkyl, -OCH2CF3, -O(CH2) cyclopropyl, -OCF3, -OCHF2, or -O(C2-C 10 ) alkenyl, -O(CH2)3C≡CH; and / or (ix) R 10 is H, halogen, or -O(C1-C2)alkyl; and / or (x) where R 9 and R 10 The two adjacent remaining parts of forming a ring bonded to the basic aromatic ring of formula (II) according to TIFF2024527473000029.tif27145; In the formula, T 1 and T 2 is independently selected from the group consisting of H, -methyl and F; wherein each hydrogen in formulae (III) to (IX) may be substituted with methyl; Het is O; G is selected from CH; R 38 are independently H, -(C1 to C 10 ) alkyl; (xi)R 11 is H or -OCH3; and / or (xii) R is H, C1-C6 alkyl, or benzyl, most preferably H; and / or (xiii) J1 to J4 are C or N.

[0067] A selection of compounds within the scope of the present invention are listed in the table below.

[0068] [Table 1] TIFF2024527473000031.tif224142TIFF2024527473000032.tif217142TIFF2024527473000033.tif243161TIFF20245274730 00034.tif250161TIFF2024527473000035.tif250161TIFF2024527473000036.tif243161TIFF2024527473000037.tif135162

[0069] The present invention is further directed to compounds according to Formula (I), (II) or according to Table 1 for use in medicine.

[0070] The present invention is further directed to the compounds according to formula (I), (II) or according to Table 1 for use in the treatment of fibrosis and neoplasms, preferably fibrosis or neoplasms located in the heart, lung, renal tract, liver, skin, pleura and retroperitoneum, more preferably, the fibrosis is selected from pleural fibrosis, retroperitoneal fibrosis, atrial fibrillation, myocardial interstitial fibrosis, idiopathic pulmonary fibrosis (IPF), interstitial lung disease, chronic kidney disease, non-alcoholic fatty liver disease, skin scars, keloids, tumor-associated fibrotic responses.

[0071] The present invention is further directed to compounds according to Formula (I), (II) or according to Table 1 for use in the treatment of inflammatory diseases such as arthritic disease, familial Mediterranean fever and pericarditis.

[0072] Compounds of the present invention that contain a basic functional group can form salts with various inorganic or organic acids. Exemplary inorganic and organic acids / bases, as well as exemplary acid / base addition salts of compounds of the present invention, are provided in the definition of "Pharmaceutically Acceptable Salts" in the "Pharmaceutical Compositions" section below. Compounds of the present invention that contain an acidic functional group can form salts with various inorganic or organic bases. Compounds of the present invention that contain both basic and acidic functional groups can be converted into either base or acid addition salts. The neutral forms of the compounds of the present invention can be regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner.

[0073] Synthesis of the compounds of the present invention Many of the compounds of the present invention are commercially available. Compounds that are not commercially available can generally be obtained as follows, and specific examples for the preparation of the compounds of the present invention are described in the Examples section.

[0074] As shown in Scheme 1, the compounds of the present invention can be synthesized by amide formation between the respective aromatic acid and aniline. TIFF2024527473000038.tif34163

[0075] Several reagents and methods are known for the formation of amides from acids and amines. Exemplary methods and / or reagents include: (a) carbodiimide-based reagents, such as, for example, N,N'-dicyclohexylcarbodiimide (DCC), N,N'-diisopropylcarbodiimide (DIC), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), optionally in combination with a weak organic base, such as, for example, triethylamine, ethyldiisopropylamine; (b) benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate, optionally in combination with a weak organic base, such as, for example, triethylamine, ethyldiisopropylamine, etc.; (c) In a first step, a carboxylic acid is converted to the corresponding acid chloride with oxalyl chloride, optionally in the presence of dimethylformamide, followed in a second step by conversion of the acid chloride to the desired amide with aniline.

[0076] Pharmaceutical Compositions Furthermore, the present invention is directed to pharmaceutical compositions comprising the compounds described above and at least one carrier.

[0077] A "pharmaceutical composition" refers to one or more active ingredients, one or more inactive ingredients that make up the carrier, and any product that results directly or indirectly from the combination, complexation, or aggregation of any two or more of the ingredients, or from the dissociation of one or more of the ingredients, or from another type of reaction or interaction of one or more of the ingredients. Thus, the pharmaceutical composition of the present invention encompasses any composition made by mixing a compound of the present invention with a pharma- ceutical acceptable carrier.

[0078] "Carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, including, but not limited to, peanut oil, soybean oil, mineral oil, sesame oil, and the like. Water is a preferred carrier when the pharmaceutical composition is administered orally. Saline and aqueous dextrose solutions are preferred carriers when the pharmaceutical composition is administered intravenously. Saline and aqueous dextrose and glycerol solutions are preferably utilized as liquid carriers for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, nonfat dry milk, glycerol, propylene, glycol, water, ethanol, and the like. The composition can contain minor amounts of wetting or emulsifying agents, or pH buffering agents, if desired. These compositions may take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained release formulations, and the like. The compositions may be formulated as suppositories with conventional binders and carriers, such as triglycerides. Oral formulations may contain standard carriers, such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, and the like. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by EW Martin. Such compositions will contain a therapeutically effective amount of the therapeutic agent, preferably in purified form, together with a suitable amount of carrier, so as to provide the form for proper administration to the patient. The formulation should be compatible with the mode of administration.

[0079] The composition of the present invention can be administered by various methods known in the art. As will be understood by those skilled in the art, the route and / or mode of administration will vary depending on the desired results. The active compound can be prepared with a carrier that protects the compound from rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable and biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid can be used. Methods for the preparation of such formulations are generally known to those skilled in the art. For example, see Sustained and Controlled Release Drug Delivery Systems, JR Robinson, ed., Marcel Dekker, Inc., New York, 1978.

[0080] In order to administer the compounds of the present invention by certain routes of administration, it may be necessary to coat the compounds with or co-administer them with a material to prevent their inactivation.For example, the compounds can be administered to an individual in a suitable carrier, such as liposomes or diluents.Pharmaceutically acceptable diluents include saline and aqueous buffer solutions.Liposomes include water-in-oil-in-water CGF emulsions and conventional liposomes (Strejan et al., J. Neuroimmunol. 7: 27 (1984)).

[0081] Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions, and sterile powders for extemporaneous preparation of sterile injectable solutions or dispersions.The use of such media and agents for pharmaceutically active substances is known in the art.Except where any conventional media or agent is incompatible with active compound, its use in the pharmaceutical composition of the present invention is contemplated.Auxiliary active compounds can also be incorporated into the composition.

[0082] Pharmaceutical compositions must typically be sterile and stable under the conditions of manufacture and storage. The composition may be formulated as a solution, microemulsion, liposome, or other ordered structure suitable for high drug concentration. The carrier may be, for example, a solvent or dispersion medium containing water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, etc.), and suitable mixtures thereof. Proper fluidity may be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion, and by the use of surfactants. In many cases, it will be preferable to include an isotonic agent in the composition, for example, sugar, polyalcohol, for example, mannitol, sorbitol, or sodium chloride. Prolonged absorption of the injectable composition may be brought about by including an agent that delays absorption, for example, monostearate salts and gelatin in the composition.

[0083] Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in the appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by sterilization microfiltration.

[0084] Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle containing a basic dispersion medium and the required other ingredients from those listed above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying (lyophilization) which produce a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

[0085] The dosage regimen is adjusted to provide the optimum desired response (e.g., therapeutic response). For example, it may be a single bolus administration, or several divided doses may be administered over time, or the dose may be relatively decreased or increased as indicated by the exigencies of the therapeutic situation. It is particularly advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. As used herein, dosage unit form refers to a physically discrete unit suitable as a unitary dosage for the individual to be treated; each unit contains a predetermined amount of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specifications for each dosage unit form of the present invention are determined and directly depend on (a) the unique properties of the active compound and the specific therapeutic effect to be achieved, and (b) the limitations inherent in the technology of compounding such active compound for the treatment of susceptibility in an individual.

[0086] Examples of pharma- ceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal chelators, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0087] For therapeutic / pharmaceutical formulations, compositions of the present invention include those suitable for enteral administration (e.g., oral or rectal) or parenteral administration (e.g., nasal, topical (including vaginal, buccal and sublingual)). The compositions may conveniently be presented in unit dosage form and may be prepared by any method known in the art of pharmacy. The amount of active ingredient (particularly the amount of a compound of the present invention) that may be combined with a carrier material to produce a pharmaceutical composition (e.g., a single dosage form) will vary depending on the individual treated and the particular mode of administration. The amount of active ingredient that may be combined with a carrier material to produce a single dosage form will generally be that amount of the composition which produces a therapeutic effect.

[0088] Generally, out of 100% (in the case of a pharmaceutical formulation / composition), the amount of active ingredient (particularly the amount of a compound of the present invention, optionally combined with other therapeutically active agents, if present in a pharmaceutical formulation / composition) will range from about 0.01% to about 99%, preferably from about 0.1% to about 70%, and most preferably from about 1% to about 30%, with the remainder preferably consisting of one or more pharma- ceutically acceptable excipients.

[0089] The amount of active ingredient, e.g., a compound of the invention, in a unit dosage form and / or when administered to an individual or used in a treatment regimen can range from about 0.1 mg to about 1000 mg (e.g., about 1 mg to about 500 mg, e.g., about 10 mg to about 200 mg) per unit, administration, or treatment regimen. In certain embodiments, suitable amounts of such active ingredients can be calculated using the mass or body surface area of ​​the individual, and can range from about 1 mg / Kg to 10 mg / Kg (e.g., about 2 mg / Kg to 5 mg / Kg), or about 1 mg / m 2 ~about 400mg / m 2 (For example, about 3 mg / m 2 ~about 350mg / m 2 or about 10 mg / m 2 ~about 200mg / m 2 ).

[0090] Compositions of the present invention suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams, or spray formulations containing carriers known in the art to be appropriate. Dosage forms for topical or transdermal administration of compositions of the present invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active compound may be mixed under sterile conditions with a pharma- ceutically acceptable carrier, and with any preservatives, buffers, or propellants which may be required.

[0091] As used herein, the phrase "enteral administration" and "administered enterally" means that the administered drug is taken up by stomach and / or intestine. Examples of enteral administration include oral and rectal administration. As used herein, the phrase "parenteral administration" and "administered parenterally" means a mode of administration other than enteral administration, usually by injection or topical application, including but not limited to intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraosseous, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, intracerebral, intraventricular, subarachnoid, intraspinal, epidural and intrasternal administration (such as by injection and / or infusion) and topical administration (e.g., on the skin, by inhalation, or transmucosal (such as buccal, sublingual or vaginal)).

[0092] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0093] These compositions may also contain auxiliary agents such as preservatives, wetting agents, emulsifiers, pH buffering agents, and dispersing agents. Prevention of the presence of microorganisms can be ensured both by sterilization procedures and by including various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol sorbic acid, etc. It may also be desirable to include isotonic agents in the composition, such as sugars, sodium chloride, etc. In addition, the absorption of injectable pharmaceutical forms can be prolonged by including agents that delay absorption, such as aluminum monostearate and gelatin.

[0094] Regardless of the route of administration selected, the compounds of the present invention, which may be used in a suitable hydrated form, and / or the pharmaceutical compositions of the present invention, are formulated into pharma- ceutically acceptable dosage forms by conventional methods known to those of skill in the art (see, e.g., Remington, "The Science and Practice of Pharmacy" edited by Allen, Loyd V., Jr., 22nd edition, Pharmaceutical Sciences, September 2012; Ansel et al., "Pharmaceutical Dosage Forms and Drug Delivery Systems", 7th edition, Lippincott Williams & Wilkins Publishers, 1999).

[0095] The actual dosage level of the active ingredient in the pharmaceutical compositions of the present invention may vary for each particular patient, composition, and mode of administration to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response and is not toxic to the patient. The dosage level selected will depend on a variety of pharmacokinetic factors, including the activity of the particular composition of the present invention employed, the route of administration, the time of administration, the rate of excretion of the particular compound employed, the duration of treatment, other drugs, compounds and / or materials used in combination with the particular composition employed, the age, sex, weight, condition, general health and medical history of the patient being treated, and similar factors well known in the medical arts.

[0096] A physician or veterinarian of ordinary skill in the art can easily determine and prescribe the effective amount of pharmaceutical composition required. For example, a physician or veterinarian can start with a dose of the compound of the invention employed in the pharmaceutical composition at a level lower than that required to achieve the desired therapeutic effect, and gradually increase the dose until the desired effect is achieved. Generally, a suitable daily dose of the composition of the invention will be the amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective amount generally varies depending on the factors listed above. Administration is preferably oral, intravenous, intramuscular, intraperitoneal, or subcutaneous, and is preferably administered proximal to the target site. If desired, the effective daily dose of the pharmaceutical composition can be administered as 2, 3, 4, 5, 6 or more divided doses administered separately at appropriate intervals throughout the day, optionally in unit dosage form. Although it is possible to administer the compound of the invention alone, it is preferred to administer the compound as a pharmaceutical formulation / composition.

[0097] In one embodiment, the compound or composition of the present invention may be administered by infusion, preferably slow continuous infusion over an extended period of time, for example, more than 24 hours, to reduce toxic side effects. Administration may also be by continuous infusion over 2-24 hours, for example, 2-12 hours. Such a regimen may be repeated one or more times as needed, for example, after 6 or 12 months.

[0098] In yet another embodiment, a compound or composition of the invention is administered by maintenance therapy, such as, for example, once a week for a period of six months or more.

[0099] For oral administration, the pharmaceutical compositions of the invention may take the form of, for example, tablets or capsules prepared by conventional means with pharma- ceutically acceptable excipients, such as binders (e.g., pregelatinized maize starch, polyvinylpyrrolidone, hydroxypropylmethylcellulose), fillers (e.g., lactose, microcrystalline cellulose, calcium hydrogen phosphate), lubricants (e.g., magnesium stearate, talc, silica), disintegrants (e.g., potato starch, sodium starch glycolate), or wetting agents (e.g., sodium lauryl sulfate). Liquid preparations for oral administration may be in the form of, for example, solutions, syrups, or suspensions, or may be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations may be prepared by conventional means with pharma- ceutically acceptable additives, such as suspending agents (e.g., sorbitol, syrups, cellulose derivatives, hydrogenated edible fats), emulsifying agents (e.g., lecithin, acacia), non-aqueous vehicles (e.g., almond oil, oily esters, ethyl alcohol, fractionated vegetable oils), preservatives (e.g., methyl- or propyl-p-hydroxycarbonate, sorbic acid), and the like. Preparations may also contain buffer salts, flavoring, coloring, and sweetening agents as deemed appropriate. Preparations for oral administration may be suitably formulated to give controlled release of the pharmaceutical compositions of the present invention.

[0100] The pharmaceutical composition may be formulated as a suppository, with traditional binders and carriers such as triglycerides. Oral formulations can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc.

[0101] For administration by inhalation, the pharmaceutical composition of the present invention is conveniently delivered in the form of an aerosol spray presentation from a pressurized pack or nebulizer with the use of a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, nitrogen, or other suitable gas). In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges of, for example, gelatin for use in an inhaler or insufflator can be formulated containing a powder mix of the pharmaceutical composition of the present invention and a suitable powder base, such as lactose or starch.

[0102] The pharmaceutical compositions of the present invention may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Injectable preparations may be provided in unit dosage form (e.g., in vials or in multi-dose containers) with the addition of a preservative. The pharmaceutical compositions of the present invention may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles and may contain formulating agents such as suspending, stabilizing or dispersing agents. Alternatively, the agent may be in powder form for constitution with a suitable vehicle (e.g., sterile pyrogen-free water) before use. Typically, compositions for intravenous administration are solutions in sterile isotonic aqueous buffer. If necessary, the composition may also include a solubilizing agent and a local anesthetic such as lignocaine to ease pain at the site of the injection. Generally, the ingredients are supplied, either individually or mixed in unit dosage form, for example, as a dry lyophilized powder or water-free concentrate in an airtight container, such as an ampule or sachet indicating the quantity of active agent. When the composition is administered by injection, it may be dispensed with an infusion bottle containing sterile pharmaceutical grade water or saline. Where the composition is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients may be mixed prior to administration.

[0103] The therapeutic / pharmaceutical composition can be administered using medical devices known in the art.For example, in a preferred embodiment, the therapeutic / pharmaceutical composition of the present invention can be administered using needleless hypodermic injection devices, such as those disclosed in US 5,399,163; US 5,383,851; US ​​5,312,335; US 5,064,413; US 4,941,880; US 4,790,824; or US 4,596,556. Examples of well-known implants and modules useful in the present invention include those described in US 4,487,603 which discloses an implantable microinfusion pump for dispensing drugs at a controlled rate, US 4,486,194 which discloses a therapeutic device for administering drugs through the skin, US 4,447,233 which discloses a drug infusion pump for delivering drugs at precise infusion rates, US 4,447,224 which discloses a variable flow rate implantable infusion device for continuous drug delivery, US 4,439,196 which discloses an osmotic drug delivery system having multiple chamber compartments, and US 4,475,196 which discloses an osmotic drug delivery system.

[0104] Many other such implants, delivery systems, and modules are known to those skilled in the art. In certain embodiments, the compounds of the present invention can be formulated to ensure proper distribution in vivo. For example, the blood-brain barrier (BBB) ​​excludes many highly hydrophilic compounds. To ensure that the compounds of the present invention cross the BBB (if desired), they can be formulated, for example, in liposomes. For methods of producing liposomes, see, for example, US 4,522,811; US ​​5,374,548; and US 5,399,331. Liposomes can contain one or more moieties that selectively transport to specific cells or organs, thus improving targeted drug delivery (see, for example, VV Ranade (1989) J. Clin. Pharmacol. 29: 685). Exemplary targeting moieties include folate or biotin (see, e.g., Low et al., U.S. Pat. No. 5,416,016), mannosides (Umezawa et al., (1988) Biochem. Biophys. Res. Commun. 153: 1038); antibodies (PG Bloeman et al. (1995) FEBS Lett. 357: 140; M. Owais et al. (1995) Antimicrob. Agents Chemother. 39: 180); and surfactant protein A receptor (Briscoe et al. (1995) Am. J. Physiol. 1233: 134).

[0105] In one embodiment of the present invention, the compound of the present invention is formulated into liposome.In a more preferred embodiment, the liposome comprises a targeting moiety.In the most preferred embodiment, the compound in liposome is delivered by bolus injection to a site proximal to the desired area.Such liposome-based composition should be fluid enough to be easily syringable, stable under the manufacturing and storage conditions, and protected from the contaminating action of microorganisms such as bacteria and fungi.

[0106] A "therapeutically effective dose" for a treatment / treatment may be measured by either complete or partial objective remission. Complete remission (CR) is defined as the absence of clinical, radiological, or other evidence of the condition, disorder, or disease. Partial remission (PR) is achieved by greater than 50% reduction in disease. The mean time to progression is a measure that characterizes the durability of objective remission.

[0107] A "therapeutically effective dose" for treatment / treatment may also be measured by its ability to keep the progression of a condition, disorder or disease constant. The ability of a compound to inhibit, reduce or ameliorate non-apoptotic cell death and / or reduce oxidative stress may be evaluated in an appropriate animal model system, such as one or more of those described below. Alternatively, these properties of the compounds of the present invention may be evaluated by examining the ability of the compound to use in vitro assays known to those of skill in the art, such as one or more of those described below. A therapeutically effective amount of a compound of the present invention may cure, heal, alleviate, ameliorate, alter, treat, ameliorate, improve or affect a condition, disorder or disease, or a symptom of a condition, disorder or disease, or a predisposition to a condition, disorder or disease in an individual. One of skill in the art will be able to determine such amounts based on factors such as the size of the individual, the severity of the individual's symptoms, and the specific composition or route of administration selected.

[0108] Injectable compositions should be sterile and fluid to the extent that the composition can be delivered by syringe. In addition to water, the carrier can be an isotonic buffered saline solution, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof.

[0109] The pharmaceutical compositions of the present invention may, if desired, be presented in a pack or dispenser device which may contain one or more unit dosage forms containing the agents. The pack may, for example, comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration.

[0110] The pharmaceutical compositions of the present invention may be administered as the sole active agent or may be administered in combination with other therapeutic and / or cosmetic active agents.

[0111] Further, the present invention relates to a method for producing a method for manufacturing a semiconductor device comprising the steps of: (a) culturing adherent cells that deposit at least one protein in the presence of at least one test compound; (b) staining of at least one protein deposited by adherent cells; (c) fixation of the adherent cells and the at least one protein; (d) microscopic detection of the at least one stained deposited protein signal; (e) data analysis of the signals detected in step (d), comprising quantifying the amount of said at least one protein deposited in the presence of said at least one test compound. wherein step (b) is performed before step (c).

[0112] In step (a), the cells are cultured in adherent cell culture. The cells may be cultured in adherent cell culture using any conditions known to the skilled artisan suitable for culturing the respective cell type. Optionally, the cells are starved for preferably 5 to 48 hours, more preferably 10 to 30 hours, most preferably 20 to 26 hours prior to applying step (a).

[0113] Preferably, the adherent cells are primary cells. In one embodiment, the adherent cells are primary human cells from patients, most preferably human lung fibroblasts. In another embodiment, the cells are primary animal cells or any adherent immortalized cells.

[0114] Preferably, at least one protein in step (a) is an extracellular matrix protein. More preferably, at least one extracellular matrix protein is selected from the group consisting of type 5 collagen, type 1 collagen, and fibulin 1. In further embodiments, more than one protein is deposited. For example, at least two, at least three, or at least four proteins are deposited.

[0115] Preferably, step (a) is carried out for at least 24 hours, more preferably from 60 to 90 hours, particularly preferably from 65 to 80 hours, most preferably from 70 to 75 hours.

[0116] At least one test compound is preferably a compound that is expected to inhibit the deposition of at least one protein.The purpose of the assay is to identify the compound that is likely to inhibit the deposition of at least one protein, and to quantify the degree of inhibition by each test compound.Optionally, at least one test compound in step a) is a small molecule; and / or an oligonucleotide, a peptide, a protein, a protac, an anticarin, an antibody, or a CRISPR.

[0117] Optionally, in step (a) at least one growth factor is present, preferably at least one transforming growth factor beta (TGFβ), more preferably TGFβ1.

[0118] In one embodiment, the staining in step (b) comprises binding of at least one antibody to the at least one protein or to at least one probe bound to the at least one protein.

[0119] Optionally, the antibody or probe comprises at least one detectable label directly conjugated to the antibody, and, optionally, the detectable label has fluorescent properties, preferably the detectable label is a fluorophore selected from AlexaFluor 488, AlexaFluor 555, AlexaFluor 637; AlexaFluor 647, AlexaFluor 568, AlexaFluor 568 and / or Qdot.

[0120] In a further embodiment, the staining in step (b) comprises binding of at least one first antibody (FA) to the at least one protein, followed by binding of the at least one first antibody (FA) with at least one secondary antibody (SA), wherein the at least one second antibody (SA) comprises at least one detectable label conjugated to the antibody, and wherein, optionally, the detectable label has fluorescent properties, preferably the detectable label is a fluorophore selected from AlexaFluor 488, AlexaFluor 555, AlexaFluor 637; AlexaFluor 647, AlexaFluor 568, AlexaFluor 568 and / or Qdot.

[0121] Optionally, in step (b), there is at least one additional co-stain, selected from the group consisting of a cell nucleus stain, a live / dead cell stain, a myofibroblast marker (e.g., αSMA stain), an apoptosis marker (e.g., caspase 3 / 7 stain).

[0122] In step (c), fixation may be performed using any reagent suitable for the purpose and known to those skilled in the art. The respective conditions are known in the art. Exemplary conditions used in certain embodiments of the present invention are 4% PFA at 37° C. for 30 minutes, or 100% methanol at −20° C. for 2 minutes. Staining in step (b) is performed before fixation in step (b).

[0123] Preferably, in step (d), 2D, 3D or 4D imaging is performed. More preferably, step (d) is performed by conventional or confocal imaging equipment.

[0124] The data analysis in step (e) may involve using a machine learning model, such as a neural network. EXAMPLES

[0125] Example of the invention 1. Compound synthesis Example 1: (E)-N-(2-butoxyphenyl)-3-(4-(cyclopropylmethoxy)phenyl)acrylamide TIFF2024527473000039.tif24128

[0126] (a) (E)-N-(2-butoxyphenyl)-3-(4-hydroxyphenyl)acrylamide (E)-3-(4-hydroxyphenyl)acrylic acid (120 mg, 0.73 mmol) was dissolved in 2 mL of dry DMF. The solution was cooled in an ice bath and 2-butoxyaniline (0.87 mmol, 1.2 eq) was added followed by a solution of benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate (1.1 mmol, 1.5 eq) in 2 mL of dichloromethane. This was followed by triethylamine (1.46 mmol, 2 eq). The mixture was stirred at 0° C. for 30 min and then at room temperature overnight. The mixture was diluted with dichloromethane and washed with 1 M aqueous hydrochloric acid, saturated sodium carbonate and brine. The organic layer was dried over magnesium sulfate and concentrated in vacuo. Purification by silica gel chromatography gave (E)-N-(2-butoxyphenyl)-3-(4-hydroxyphenyl)acrylamide (87 mg).

[0127] (b) (E)-N-(2-butoxyphenyl)-3-(4-(cyclopropylmethoxy)phenyl)acrylamide (Example 1) To a solution of (E)-N-(2-butoxyphenyl)-3-(4-hydroxyphenyl)acrylamide (1 eq), cyclopropylmethanol (2 eq), and triphenylphosphine (2 eq) in anhydrous tetrahydrofuran (15 mL) was added diisopropyl azodicarboxylate (2 eq) over 10 min at 0° C. After 30 min, the cooling bath was removed and the solution was stirred at room temperature for 16 h. The solvent was concentrated in vacuo and the remaining residue was purified by flash chromatography to give (E)-N-(2-butoxyphenyl)-3-(4-(cyclopropylmethoxy)phenyl)acrylamide in 77% yield. TIFF2024527473000040.tif51170

[0128] Example 2: (E)-N-(2-butoxyphenyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)acrylamide TIFF2024527473000041.tif22128

[0129] (a) (E)-3-(4-hydroxyphenyl)acrylate methyl ester To a solution of (E)-3-(4-hydroxyphenyl)acrylic acid (0.8 g) in methanol (40 mL) was added concentrated sulfuric acid (0.67 mL). The solution was heated to reflux for 5 h, cooled to room temperature, and then quenched by the addition of saturated aqueous sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate and the combined organic fractions were washed with water, brine, dried over magnesium sulfate, and concentrated in vacuo to give methyl (E)-3-(4-hydroxyphenyl)acrylate (800 mg), which was sufficiently pure for further transformation. TIFF2024527473000042.tif11164HLM-01-046, 90% yield.

[0130] (b) (E)-3-(4-(2,2,2-trifluoroethoxy)phenyl)methyl acrylate A dried flask was charged with NaH (1.2 equiv.) under argon atmosphere. Dry dimethyl sulfoxide (4 mL) was added to the reaction flask and stirred at 0° C. for 15 min. A solution of (E)-3-(4-(2,2,2-trifluoroethoxy)phenyl)methyl acrylate (890 mg, 5 mmol, 1 equiv.) in DMSO (2 mL) was added slowly to the suspension over 10 min. The reaction mixture was stirred at 0° C. for 30 min. 2,2,2-trifluoroethyl iodide (1.5 mL, 15 mmol, 3 equiv.) was added to the reaction flask. The reaction was then stirred at 80° C. for 24 h. After completion of the reaction, it was quenched by addition of water and extracted with ethyl acetate. The organic phase was evaporated to dryness and purified by silica gel chromatography.

[0131] (c) (E)-3-(4-(2,2,2-trifluoroethoxy)phenyl)acrylic acid To a solution of methyl (E)-3-(4-(2,2,2-trifluoroethoxy)phenyl)acrylate (1 eq.) in methanol (4 mL) was added potassium carbonate (5 eq.) dissolved in water (4 mL). The reaction mixture was refluxed for 3 h, after which the methanol was removed under reduced pressure. The solution was then cooled to 0° C. and acidified to pH 2 by addition of hydrochloric acid (1 M). The mixture was extracted with diethyl ether and the combined organic layers were washed with brine, dried over sodium sulfate and evaporated in vacuo to give the product as a white solid in 90% yield.

[0132] (d) (E)-N-(2-butoxyphenyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)acrylamide (E)-N-(2-butoxyphenyl)-3-(4-(2,2,2-trifluoroethoxy)phenyl)acrylamide was prepared from (E)-3-(4-(2,2,2-trifluoroethoxy)phenyl)acrylic acid and 2-butoxyaniline in a similar manner as described for (E)-N-(2-butoxyphenyl)-3-(4-hydroxyphenyl)acrylamide (Example 1, step (a)). The product was obtained in 63% yield. TIFF2024527473000043.tif45168

[0133] Example 3: (E)-3-(4-methoxyphenyl)-N-(2-phenoxyphenyl)acrylamide TIFF2024527473000044.tif22128

[0134] A suspension of (E)-3-(4-methoxyphenyl)acrylic acid (150 mg, 0.84 mmol, 1 eq) in dry dichloromethane (3 mL) was treated with oxalyl chloride (1.68 mmol, 2 eq) and a catalytic amount of dimethylformamide at 0° C. under argon. After 5 min, the solution was warmed and stirred at ambient temperature for 1 h. The solvent was then removed under reduced pressure to give the acid chloride as a yellow solid. A solution of the acid chloride in dry dichloromethane (4 mL) was added to a solution of 2-phenoxyaniline (1.01 mmol, 1.2 eq) and triethylamine (1.01 mmol 1.2 eq) in dichloromethane (3 mL) at 0° C. The suspension was stirred at room temperature for 16 h and quenched by the addition of water. The mixture was diluted with dichloromethane, washed with aqueous ammonium chloride, saturated sodium bicarbonate, then dried over magnesium sulfate and concentrated in vacuo. The crude product was purified by silica gel chromatography to give 139 mg of (E)-3-(4-methoxyphenyl)-N-(2-phenoxyphenyl)acrylamide in 48% yield (139 mg). TIFF2024527473000045.tif38164

[0135] Example 4: (E)-N-(4-butoxypyridin-3-yl)-3-(4-methoxyphenyl)acrylamide TIFF2024527473000046.tif22128

[0136] (a) 4-Butoxy-3-nitropyridine To a solution of 3-nitropyridin-4-ol (1.0 g, 7.13 mmol) and triphenylphosphine (2.8 g, 10.71 mmol, 1.5 eq) in anhydrous tetrahydrofuran (10 mL) were added diisopropyl azodicarboxylate (2.16 g, 10.71 mmol, 1.5 eq) in tetrahydrofuran (2.5 mL) and 1-butanol (794 mg, 10.71 mmol, 1.5 eq) in tetrahydrofuran (2.5 mL) at 0 °C simultaneously. The temperature of the reaction mixture was allowed to slowly rise to room temperature and stirring was continued overnight at room temperature. At the end of this period it was diluted with ethyl acetate (25 mL), washed with water (2 x 50 mL), dried over Na2SO4, filtered and the solvent was evaporated. The residue was purified by silica column (dichloromethane / methanol = 50 / 1) to give 4-butoxy-3-nitropyridine as a light yellow solid (400 mg). TIFF2024527473000047.tif18164

[0137] (b) 4-Butoxypyridin-3-amine A suspension of 4-butoxy-3-nitropyridine (386 mg, 2.0 mmol) in ethanol (9.2 ml) and acetic acid (0.51 ml) was heated at 60° C. After addition of iron powder (678 mg, 12.0 mmol, 6.0 eq) and iron(III) chloride hexahydrate (55 mg), the mixture was stirred under reflux for 18 h. After cooling to room temperature, the mixture was diluted with ethyl acetate (100 ml) and filtered through Celite. The filtrate and washings were combined and washed with water. The organic phase was dried over MgSO4 and then concentrated in vacuo to give 4-butoxypyridin-3-amine as a pale yellow solid (310 mg). The crude product can be used in the next step without further purification.

[0138] (c) (E)-N-(4-butoxypyridin-3-yl)-3-(4-methoxyphenyl)acrylamide (Example 4) A suspension of (E)-3-(4-methoxyphenyl)but-2-enoic acid (1.4 mmol, 250 mg) in dry dichloromethane (5 mL) was treated with oxalyl chloride (1.5 eq) and catalytic amount of dimethylformamide (2 drops) under argon atmosphere at 0°C. After 5 min, the solution was warmed to room temperature and stirred at ambient temperature for 1 h. The solvent was removed under reduced pressure to give the acid chloride as a yellow solid. A solution of the acid chloride in additionally dried dichloromethane (7 mL) was added to a solution of 4-butoxypyridin-3-amine (300 mg, 1.2 eq) and triethylamine (1.2 eq) in dichloromethane (5 mL) at 0°C. The suspension was stirred at ambient temperature for 4 h and quenched with water. The mixture was diluted with dichloromethane, washed with aqueous ammonium chloride, saturated sodium bicarbonate, and then dried over MgSO4. After concentration in vacuo, the residue was purified by silica gel column (dichloromethane / methanol 15:1) to give (E)-N-(4-butoxypyridin-3-yl)-3-(4-methoxyphenyl)acrylamide as a white solid (530 mg). TIFF2024527473000048.tif45169

[0139] Example 5: (E)-N-(2-((4-fluorobenzyl)oxy)phenyl)-3-(4-methoxyphenyl)acrylamide TIFF2024527473000049.tif23128

[0140] (a) 1-((4-fluorobenzyl)oxy)-2-nitrobenzene To a solution of 2-nitrophenol in acetonitrile was added anhydrous potassium carbonate. The mixture was stirred at room temperature for 30 minutes. Catalytic amounts of potassium iodide and 1-(bromomethyl)-4-fluorobenzene were added. The reaction was stirred at 75° C. for 8 hours. Water (125 mL) was added to the reaction mixture and the reaction mixture was extracted with ethyl acetate. The combined organic layers were washed with brine and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure and the residue was purified by flash chromatography. TIFF2024527473000050.tif12164

[0141] (b) 2-((4-fluorobenzyl)oxy)aniline To a solution of 1-((4-fluorobenzyl)oxy)-2-nitrobenzene (1 eq) in methanol (7.5 mL) and water (7.5 mL) was added ammonium chloride (10 eq) and iron powder (5 eq). The reaction mixture was stirred at 90° C. for 16 h. The mixture was filtered and water (50 mL) was added to the filtrate. The filtrate was extracted with ethyl acetate and the combined organic layers were extracted with brine, dried over magnesium sulfate and concentrated in vacuo to give crude 2-((4-fluorobenzyl)oxy)aniline (80% yield), which was used without purification for further transformations. TIFF2024527473000051.tif38164

[0142] (c) (E)-N-(2-((4-fluorobenzyl)oxy)phenyl)-3-(4-methoxyphenyl)acrylamide (Example 5) (E)-3-(4-Methoxyphenyl)acrylic acid and 2-((4-fluorobenzyl)oxy)aniline were reacted in a similar manner as described for the synthesis of (E)-3-(4-methoxyphenyl)-N-(2-phenoxyphenyl)acrylamide (Example 3) to provide (E)-N-(2-((4-fluorobenzyl)oxy)phenyl)-3-(4-methoxyphenyl)acrylamide in 57% yield. TIFF2024527473000052.tif38164

[0143] Example 6: (E)-3-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-N-(2-methoxyphenyl)acrylamide TIFF2024527473000053.tif22128

[0144] (a) (E)-3-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)acrylic acid A mixture of 2.2 g (11.8 mmol) of 2,2-difluoro-benzo[1,3]dioxole-5-carbaldehyde, 2.71 g (26.0 mmol) of malonic acid, 0.2 g (2.4 mmol) of piperidine, and 9 ml of pyridine was maintained at reflux temperature until carbon dioxide evolution ceased (3 h). After cooling to room temperature, the reaction mixture was poured into 100 g of ice and 30 ml of 6N HCl. The precipitate was isolated, washed with water, and dried to give the desired product HLM1319 (2.53 g, 11.1 mmol, 94% yield) as a white solid, which was used in the next step without further purification.

[0145] (b) (E)-3-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-N-(2-methoxyphenyl)acrylamide (Example 6) Following the synthesis procedure of Example 3 ((E)-3-(4-methoxyphenyl)-N-(2-phenoxyphenyl)acrylamide), (E)-3-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-N-(2-methoxyphenyl)acrylamide was obtained as a brown solid (400 mg, 1.2 mmol, 68% yield) from (E)-3-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)acrylic acid (400 mg, 1.75 mmol) after purification by silica gel chromatography (PE / EA=8 / 1). TIFF2024527473000054.tif45168

[0146] The following examples, listed in Table 2 below, were obtained using methods similar to those used for the synthesis of Examples 1-6.

[0147] [Table 2] TIFF2024527473000056.tif230148TIFF2024527473000057.tif230162TIFF2024527 473000058.tif230152TIFF2024527473000059.tif230144TIFF2024527473000060.t if230148TIFF2024527473000061.tif230148TIFF2024527473000062.tif230151TIF F2024527473000063.tif230150TIFF2024527473000064.tif230137TIFF2024527473 000065.tif230148TIFF2024527473000066.tif230137TIFF2024527473000067.tif2 30157TIFF2024527473000068.tif230144TIFF2024527473000069.tif230146TIFF20 24527473000070.tif230133TIFF2024527473000071.tif230157TIFF2024527473000 072.tif230157TIFF2024527473000073.tif230152TIFF2024527473000074.tif23035

[0148] Example 66: (E)-3-(4-(3-(1-benzyl-1H-1,2,3-triazol-4-yl)propoxy)phenyl)-N-(2-butoxyphenyl)acrylamide TIFF2024527473000075.tif23128

[0149] (Azidomethyl)benzene (77 mg, 0.58 mmol) and (E)-N-(2-butoxyphenyl)-3-(4-(pent-4-yn-1-yloxy)phenyl)acrylamide (220 mg, 0.58 mmol) (1.00 equiv.) were dissolved in dichloromethane. To this solution, a freshly prepared aqueous solution of copper(II) sulfate pentahydrate (12 mg, 0.047 mmol, 0.08 equiv.) and sodium ascorbate (23 mg, 0.12 mmol, 0.20 equiv.) was added while the mixture was vigorously stirred. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction mixture was extracted with dichloromethane and the organic phase was dried over magnesium sulfate. After the organic layer was evaporated to dryness, the residue was purified by silica gel chromatography (PE / EA=1 / 1) to give (E)-3-(4-(3-(1-benzyl-1H-1,2,3-triazol-4-yl)propoxy)phenyl)-N-(2-butoxyphenyl)acrylamide (153 mg, 0.3 mmol, 51%) as a white solid. TIFF2024527473000076.tif65170

[0150] Example 67: E) -N-(2-butoxyphenyl)-3-(3,4-dimethoxyphenyl)-N-methylacrylamide TIFF2024527473000077.tif23128

[0151] (E)-N-(2-butoxyphenyl)-3-(3,4-dimethoxyphenyl)acrylamide (Example 84, 1 eq) was added to a suspension of sodium hydride (1.4 eq) in dry THF (8 mL). The mixture was stirred for 5 min until hydrogen evolution ceased. Methyl iodide (1.4 eq) was added and stirring was continued for 16 h. The mixture was poured into ether, washed with brine, dried over magnesium sulfate and evaporated to dryness in vacuo. Purification of the crude material by flash chromatography afforded (E)-N-(2-butoxyphenyl)-3-(3,4-dimethoxyphenyl)-N-methylacrylamide in 76% yield. TIFF2024527473000078.tif45169

[0152] The following compounds, listed in Table 3 below, may be prepared in a similar manner as described for Example 67.

[0153] [Table 3] TIFF2024527473000080.tif221143TIFF2024527473000081.tif221129

[0154] Example 78: (E)-N-(2-butoxyphenyl)-3-(4-methoxyphenyl)-2-methylacrylamide TIFF2024527473000082.tif20128

[0155] (a) (E)-3-(4-methoxyphenyl)-2-methylacrylic acid ethyl ester To a solution of p-anisaldehyde (32 g, 0.24 mol) and ethyl (2-dimethoxyphosphinyl)-2-propanoate (58.8 g, 0.28 mol) in toluene (160 mL) was added NaOtBu (33.6 g, 0.35 mol) at 0° C. over 30 min under N2 atmosphere. The reaction mixture was warmed to room temperature and stirred for 15 min. After completion of the reaction, the reaction mixture was neutralized with 10% aqueous HCl. The toluene layer was separated, washed with water, dried over sodium sulfate, and concentrated in vacuum. The residue was purified by silica gel chromatography to give 22.7 g of ethyl (E)-3-(4-methoxyphenyl)-2-methylacrylate. TIFF2024527473000083.tif12164

[0156] (b) (E)-3-(4-methoxyphenyl)-2-methylacrylic acid A 50 mL round-bottom flask was charged with ethyl (E)-3-(4-methoxyphenyl)-2-methylacrylate (10 mmol, 1 eq) and LiOH·H2O (50 mmol, 5 eq) in a mixture of tetrahydrofuran / water (1:1, 0.25 M). The reaction flask was heated to 80 °C and stirred for 3 h. After cooling to room temperature, the mixture was extracted with diethyl ether. The aqueous phase was acidified by the addition of 2N hydrochloric acid and extracted with ethyl acetate. The combined organic layers were dried over magnesium sulfate and subsequently evaporated to dryness (18.6 g).

[0157] (c) (E)-N-(2-butoxyphenyl)-3-(4-methoxyphenyl)-2-methylacrylamide (Example 78) Following the synthesis procedure of Example 3 ((E)-3-(4-methoxyphenyl)-N-(2-phenoxyphenyl)acrylamide), (E)-3-(4-methoxyphenyl)-2-methylacrylic acid and 2.butoxyaniline were utilized to obtain (E)-N-(2-butoxyphenyl)-3-(4-methoxyphenyl)-2-methylacrylamide in 92% yield. TIFF2024527473000084.tif38169 The examples listed in Table 4 below were obtained in a similar manner as described for Example 78.

[0158] [Table 4] TIFF2024527473000086.tif21645

[0159] Example 167. (E)-N-(2-(2-(3-methyl-3H-diazirin-3-yl)ethoxy)phenyl)-3-(4-(prop-2-yn-1-yloxy)phenyl)acrylamide TIFF2024527473000087.tif48128

[0160] (a) (E)-3-(4-(prop-2-yn-1-yloxy)phenyl)acrylic acid methyl ester To a solution of (E)-3-(4-hydroxyphenyl)methyl acrylate (2.8 mmol, 500 mg, 1.0 equiv.) and propargyl bromide (3.4 mmol, 401 mg, 1.2 equiv.) in acetone (5.6 mL) was added K2CO3 (3.4 mmol, 465 mg, 1.2 equiv.) and the mixture was stirred at 60 °C for 24 h. The reaction mixture was cooled to room temperature, filtered to remove solids, and volatiles were removed under reduced pressure. The crude product was purified by column chromatography on silica gel to give (E)-3-(4-(prop-2-yn-1-yloxy)phenyl)methyl acrylate (573 mg, 94%). TIFF2024527473000088.tif28163

[0161] (b) (E)-3-(4-(prop-2-yn-1-yloxy)phenyl)acrylic acid Methyl (E)-3-(4-(prop-2-yn-1-yloxy)phenyl)acrylate (573 mg, 2.7 mmol) was dissolved in methanol (10 ml). 1 M aqueous sodium hydroxide solution (10 mL) was added. The reaction mixture was stirred at room temperature for 2 h. The product was acidified (pH=3) with 2N HCl and the aqueous layer was extracted with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 510 mg of (E)-3-(4-(prop-2-yn-1-yloxy)phenyl)acrylic acid (yield: 95%, white solid), which was used in the next step without purification.

[0162] (c) (E)-N-(2-hydroxyphenyl)-3-(4-(prop-2-yn-1-yloxy)phenyl)acrylamide (E)-3-(4-(prop-2-yn-1-yloxy)phenyl)acrylic acid (293 mg, 1 mmol), DIPEA (1 mmol, 174 μL) and HATU (418 mg, 1.1 mmol) were added in 10 mL of DCM at 0° C. and stirred for 30 min. Then 2-aminophenol (2 mmol, 218 mg) was added and stirring was continued overnight. After quenching with 1M HCl, it was extracted with ethyl acetate and the combined organic phase was dried over Na2SO4. After concentration in vacuum, the residue was purified by silica column to give (E)-N-(2-hydroxyphenyl)-3-(4-(prop-2-yn-1-yloxy)phenyl)acrylamide (270 mg, 92%). TIFF2024527473000089.tif28156

[0163] (d) (E)-N-(2-(2-(3-methyl-3H-diazirin-3-yl)ethoxy)phenyl)-3-(4-(prop-2-yn-1-yloxy)phenyl)acrylamide (HLM-01-543) To a mixture of (E)-N-(2-hydroxyphenyl)-3-(4-(prop-2-yn-1-yloxy)phenyl)acrylamide (0.27 mmol, 80 mg), triphenylphosphine (1.2 eq, 0.33 mmol, 86 mg) and (3-methyl-3H-diazirin-3-yl)ethanol [prepared according to ref. 42] (1.5 eq, 0.41 mmol, 41 mg) in THF (3 ml) was added diethyl azodicarboxylate (1.2 eq, 0.33 mmol, 51 μL) at 0 °C. After the solution was stirred at room temperature overnight, the solvent was removed in vacuo and the residue was purified by silica column to give (E)-N-(2-(2-(3-methyl-3H-diazirin-3-yl)ethoxy)phenyl)-3-(4-(prop-2-yn-1-yloxy)phenyl)acrylamide (HLM-01-543, 37 mg, 36%). TIFF2024527473000090.tif52160

[0164] Example 168: (E)-N-(2-(2-(3-methyl-3H-diazirin-3-yl)ethoxy)phenyl)-3-(4-(pent-4-yn-1-yloxy)phenyl)acrylamide TIFF2024527473000091.tif48128

[0165] Following the procedure for the synthesis of (E)-N-(2-(2-(3-methyl-3H-diazirin-3-yl)ethoxy)phenyl)-3-(4-(prop-2-yn-1-yloxy)phenyl)acrylamide (HLM-01-543), (E)-N-(2-(2-(3-methyl-3H-diazirin-3-yl)ethoxy)phenyl)-3-(4-(pent-4-yn-1-yloxy)phenyl)acrylamide (HLM-02-544) was obtained as a white solid (51 mg, 53%). TIFF2024527473000092.tif59161

[0166] 2. Screening Assays material and method primary cell culture Primary human lung fibroblasts (phLF) were isolated by outgrowth from human lung tissue derived from lung explants or tumor-free areas of lung resections as previously described (29, 30). Cells were cultured in Dulbecco's modified Eagle's medium F-12 supplemented with 20% (v / v) specially treated fetal bovine serum (PAN Biotech, catalog number 100 International Units of penicillin per mL and 100 μg / mL streptomycin. The medium was changed every 2–3 days and cells were passaged at 80–90% confluence at a ratio of 1:5 or 1:6. Cells were used for experiments up to passage 7. For screening of ECM deposition drugs, 0.5–1 × 106 cells were expanded from passage 1 to passage 5 at a ratio of 1:6 each time. More than 100 × 106 cells were trypsinized at passage 5 and cryopreserved in 90% (v / v) fetal bovine serum and 10% (v / v) dimethyl sulfoxide. Mr. Frosty (ThermoFisher Cells were slowly frozen using a freezing container (Agilent Scientific). For reseeding, phLF was thawed in a 37°C water bath and cells were washed with medium before plating. After reaching confluence at passage 6, cells were used for ECM deposition assay. Primary human dermal fibroblasts (catalog no. DF-F) were purchased from ZenBio Inc. and cultured according to the manufacturer's instructions.

[0167] ECM deposition assay phLF were cultured in DMEM F-12 medium supplemented with 20% fetal bovine serum (FBS) and antibiotic supplements as described above. Cells were seeded at 6000 cells / well in 384-well CellCarrier plates (Perkin Elmer, Cat. No. 6007550). After overnight culture, cells were starved for 24 hours in serum-reduced medium (1% FBS with 0.1 mM 2-phosphoascorbic acid (Sigma, Cat. No. 49752)). Cells were then treated with TGFβ1 (1 ng / ml) or vehicle, followed by addition of small molecules or appropriate vehicle controls. After 72 h of culture, the medium was replaced with starvation medium supplemented with 1 μg / mL AlexaFluor-488 fluorophore-conjugated anti-collagen type 5 antibody (SantaCruz, Clone C-5, Cat. No. sc-166155 AF488), 0.66 μg / mL AlexaFluor-555 fluorophore-conjugated anti-collagen type 1 antibody (Rockland, Cat. No. 600-401-103-0.1), and 1 μg / mL AlexaFluor-637 fluorophore-conjugated anti-fibulin 1 antibody (SantaCruz, Clone C-5, Cat. No. sc-25281 AF647) and 1 μg / mL Hoechst H33342 (Sigma). Fluorescent conjugation of collagen type 1 antibody was performed using the AlexaFluor-555 Protein Labeling Kit (Invitrogen, Cat. No. A20174) according to the manufacturer's instructions. The labeling effect was controlled by photometric means.

[0168] After 4 h of incubation, cells were washed three times with PBS and fixed with paraformaldehyde (PFA). For automated liquid handling in 384-well plates, an INTEGRA Viaflo II pipette (INTEGRA, Zizers, Switzerland, Cat. No. 4642), an INTEGRA Assist Plus (INTEGRA, Zizers, Switzerland) equipped with 125 μL GripTips™ pipette tips (INTEGRA, Zizers, Switzerland, Cat. No. 6464) and a sterile reagent reservoir (INTEGRA, Cat. No. 4311) was applied. All automated pipetting steps with the INTEGRA Assist Plus were performed at 9.5 μL / s to ensure proper integrity of the deposited ECM and its attachment to the culture surface in the wells of the 384-well plate. Automated liquid handling during cell seeding was performed at 89.3 μL / s. Removal of liquid from the well plate was performed by manually inverting the plate. After fixation, automated imaging was achieved by using a confocal laser scanning microscope (LSM710, Zeiss) equipped with an automatic focus detection function for three-dimensional image acquisition (1024px x 1024px x 9px, which corresponds to dimensions of 1417 μm x 1417 μm x 16 μm). For post-acquisition analysis, images were imported into IMARIS software (Bitplane) and volume detection or alternatively quantification of mean fluorescence intensity was performed, and Hoechst-stained cell nuclei were automatically counted by using the spot detection algorithm of Imaris.

[0169] Human precision-cut lung slices (PCLS) and fibrotic cocktail treatment PCLS were prepared as described previously (31, 32). Briefly, PCLS were prepared from tumor-free peritumoral tissue. Lung tissue was inflated with 3% agarose solution and solidified at 4 °C. Tissue blocks were cut into μm-thick PCLS using a vibrating microtome Hyrax V50 (Zeiss). PCLS were cultured in DMEM F-12 medium and treated with a profibrotic cocktail (31) or vehicle, as well as small molecules or vehicle, for 7 days as described previously. Supernatants were collected after culture and treatment. PCLS were washed 500 times in PBS and proteins were extracted as described previously (33). Briefly, PCLS were pooled in Eppendorf tubes and lysed in 500 μl ice-cold RIPA buffer (50 mM Tris-Cl pH 7.4, 150 mM NaCl, 1% NP40, 0.25% sodium deoxycholate) containing 1× Roche complete mini protease inhibitor cocktail (Roche, Cat. No. 11697498001). After 2 h incubation at 4°C with rotation, lung sections were removed from the lysate and protein content was measured.

[0170] Cytotoxicity assay Viability / cytotoxicity assay kits for live and dead animal cells were obtained from Biotium, catalog number 3002). CellEvent™ Caspase 3 / 7 Green Detection Reagent was obtained from Invitrogen, catalog number C10423. For the MTT assay, thiazolyl blue tetrazolium was purchased from SigmaAldrich (M5655-1G). All of these kits and assays were used according to the manufacturer's instructions.

[0171] Antibodies and dyes for immunofluorescence The following antibodies were used for immunofluorescence microscopy: monoclonal mouse anti-collagen type 5 (1 mg / mL) from Sigma Aldrich (catalog number sc-166155), monoclonal mouse anti-collagen type 5 AlexaFluor-488 conjugate from Sigma Aldrich (catalog number sc-166-155AF488), polyclonal rabbit anti-collagen type 1 (catalog number 600-401-103-0.5) from Rockland, monoclonal mouse anti-fibrin 1 (catalog number sc-25281) from SantaCruz, monoclonal mouse anti-fibrin 1 AlexaFluor-647 conjugate (1 mg / mL) from SantaCruz (catalog number sc-25281AF647), and polyclonal rabbit anti-fibronectin (1 mg / mL) from SantaCruz (catalog number sc-9068). Hoechst-33342 was obtained from Sigma (catalog number B2261). The following secondary antibodies were used: AF488 donkey anti-mouse Ab (Invitrogen, Catalog No. A21202), AF568 donkey anti-mouse Ab (Invitrogen, Catalog No. A11004), and AF568 donkey anti-mouse Ab (Invitrogen, Catalog No. A11011). Alexa Fluor 568 phalloidin (Invitrogen, A12380) was used for immunofluorescence staining of actin stress fibers. 4',6-diamidino-2-phenylindole (DAPI) was obtained from Sigma-Aldrich (Cat. No. D9564).

[0172] immunocytochemistry For standard immunofluorescence staining, 5000 phLFs were seeded in a flat-bottom 96-well imaging plate (catalog no. 353376, BD Biosciences). After incubation, cells were fixed with either 4% PFA for 30 min at 37°C or 100% methanol for 2 min at -20°C. If necessary, phLFs were permeabilized with 0.25% (v / v) Triton X-100 in PBS for 15 min. After washing with 100 μL of PBS, blocking was performed by incubation with 5% (w / v) BSA in PBS for 1 h. Primary antibodies were diluted in 1% bovine serum albumin (BSA, Sigma) in PBS and incubated for 16 h at 4°C, followed by three washes with PBS for 20 min each. Secondary antibodies were diluted in 1% bovine serum albumin (BSA, Sigma) in PBS and incubated for 1 h at room temperature, followed by three washes with PBS for 20 min each. 4% paraformaldehyde (w / v) in phosphate buffered saline was prepared from paraformaldehyde from Sigma (catalog number 15,812-7). Bovine serum albumin was obtained from Sigma (catalog number A3059). Triton X-100 was obtained from AppliChem (catalog number A1388).

[0173] Confocal 3D and 4D Imaging Confocal time-lapse microscopy was performed on an LSM710 system (Carl Zeiss) including an inverted AxioObserver.Z1 stand with phase contrast and epi-illumination optics and operated by ZEN2009 software (Carl Zeiss). The following objectives were used for imaging: EC Plan-Neofluar 20x / 0.8NA (Carl Zeiss), LD C-Apochromat 40x / 1.1NA water immersion objective (Carl Zeiss), and LCI PLN-NEOF DICIII 63x / 1.30NA water immersion objective (Carl Zeiss). For 4D imaging, cells were maintained in an incubation chamber (Carl Zeiss) under standard culture conditions (37 °C and 5% CO2). The thickness of a single confocal layer in the Z-stack was set according to the optimal value suggested by the ZEN2009 software. Confocal data sets were either maximum intensity projected in ZEN2009 software (Carl Zeiss) and / or imported into Imaris 9.0.0-9.3.1 software (Bitplane) for analysis.

[0174] Protein isolation, SDS-PAGE, Western blotting and ELISA Cells were scraped off the plastic dishes and placed directly into 200 μl ice-cold RIPA buffer containing 1× Roche complete mini protease inhibitor cocktail. Samples were incubated on ice for 30 min, after which insoluble material was removed by centrifugation at 14.000 g for 15 min at 4°C, and the supernatant was further processed. Samples were mixed with 50 mM Tris-HCl, pH 6.8, 100 mM DTT, 2% SDS, 1% bromphenol blue, and 10% glycerol, and proteins were separated using standard SDS-10% PAGE. For immunoblotting, proteins were transferred to PVDF (Millipore (Billerica, MA, (USA)), 0.45 μm or 0.2 μm) membranes, which were blocked with 5% milk in TBST (0.1% Tween 20 in TBS) and incubated with primary antibodies followed by HRP-conjugated secondary antibodies overnight at 4°C and for 1 h at room temperature, respectively. The following primary antibodies were used for immunoblotting: monoclonal mouse anti-collagen type 5 (1 mg / mL) from Sigma Aldrich (catalog number sc-166155), polyclonal rabbit anti-collagen type 3 (1 mg / mL) from Rockland (catalog number 600-401-105), polyclonal rabbit anti-collagen type 1 (1 mg / mL) from Rockland (catalog number 600-401-103-0.5), monoclonal mouse anti-fibrin 1 (1 mg / mL) from SantaCruz (catalog number sc-25281), polyclonal rabbit anti-fibronectin (1 mg / mL) from SantaCruz (catalog number sc-9068), and monoclonal mouse anti-β-actin peroxidase (AC-15, Sigma, 1:10000). Goat anti-rabbit and goat anti-mouse IgG conjugated to horseradish peroxidase (Cell Signaling, 1:10000) were applied as secondary antibodies. CXCL / IL-8 concentrations were determined using the Human IL-8 / CXCL8 DuoSet ELISA (DY208-05) according to the manufacturer's protocol.

[0175] mRNA isolation, cDNA synthesis and qRT-PCR RNA extraction from cultured phLF was performed using the PeqGold RNA kit (Peqlab) according to the manufacturer's instructions. The concentration of isolated RNA was determined spectrophotometrically at a wavelength of 260 nm (NanoDrop1000). cDNA was synthesized with the GeneAMP PCR kit (Applied Biosystems, Foster City, CA, USA) using random hexamers, using 1 μg of isolated RNA per reaction. Denaturation was performed in an Eppendorf Mastercycler with the following settings: lid=45°C, 70°C for 10 min, and 4°C for 5 min. Reverse transcription was performed in an Eppendorf Mastercycler with the following settings: lid=105°C, 20°C for 10 min, 42°C for 60 min, and 99°C for 5 min. qRT-PCR reactions were performed in a LightCycler® 480II (Roche (Risch, Switzerland)) using SYBR Green I Master under standard conditions: 95°C for 5 min followed by 45 cycles of 95°C for 5 s (denaturation), 59°C for 5 s (annealing) and 72°C for 20 s (extension) in triplicate. Target genes were normalized to HPRT expression.

[0176] Microarray and UMAP-Regulatory Pattern Clustering (UMAP-RPC) Total RNA was isolated with the PEQGold Total RNA Kit (PeqLab) according to the manufacturer's instructions, including gDNA removal. RNA quality was assessed using an Agilent 2100 Bioanalyzer, and RNA with an RIN >7 was used for microarray analysis. Total RNA (150 ng) was amplified using the WT PLUS Reagent Kit (Thermo Fisher Scientific Inc., Waltham, USA). Amplified cDNA was hybridized on human ClariomS arrays (Thermo Fisher Scientific). Staining and scanning (GeneChip Scanner 3000 7G) were performed according to the manufacturer's instructions. The Transcriptome Analysis Console (TAC: version 4.0.0.25; Thermo Fisher Scientific) was used for quality control and to obtain annotated normalized SST-RMA gene-level data. Statistical analysis was performed using the statistical programming environment R (R Development Core Team, ref. 1). Genewise testing for expression changes was performed using paired limma t-tests and Benjamini-Hochberg multiple testing correction (FDR < 10%). To reduce background, gene sets were filtered using DABG p-values ​​< 0.05 in at least one sample per pair and at least two of the three pairs per analysis. Heatmaps were generated using GraphPad Prism v7. Regulatory pattern clustering (RPC) was based on uniform manifold approximation projection (UMAP) (35). mRNA abundance from microarray data was normalized (as shown as an example in Fig. F), and abundances of all four different conditions were summarized into linear vectors (Fig. 5E) projected into a two-dimensional space (Fig. 5G) using UMAP. Clusters of genes were then extracted. Gene / protein interactions were visualized using the String Database (www.stringdb.org).

[0177] Fibrotic Pattern Detection with Artificial Intelligence (FANTAIL) - An Inferential Classification and Detection Model for Inhibition of ECM Deposition A convolutional neural network (CNN) was trained for a complex image detection and classification task using KERAS high-level API (https: / / github.com / fchollet / keras / ) implementing TensorFlow. The CNN design (Figure 10A) followed the most accepted guidelines as in (34). The best convolutional processing was achieved with three convolutional layers, convolving the image with 24 filters per layer and pooling out the data with a 2 × 2 pooling matrix in the convolutional layers. The specific image classification and detection task was based on the detection of scattered fibrotic and cellular patterns, where the image edge patterns were frequently interrupted. In large-dimension images (1024 × 1024 × RGB), the dimensional orientation of the fibrotic patterns seemed to be randomly oriented with various shapes, sizes, and clustering. Therefore, rectified linear unit (ReLU) activation was used as the activation mode to detect pattern edges, and the adadelta optimizer was selected for an efficient CNN learning process. The image control dataset was used to train the classifier. The dataset consisted of 295 immunofluorescence images annotated as "toxic" (treated with 5% ethanol), 390 images annotated as "fibrotic" (treated with TGFβ1), and 390 images annotated as "normal" (untreated). Images annotated as "normal" were labeled as "hits", while images annotated as "toxic" and "fibrotic" were combined under the label "other". Images were randomly assigned to a training dataset (75%) and a validation dataset (25%). As images of size 1024×1024 are an unusually large input for a CNN, we aimed to test the CNN efficiency of a large np×np subset of each image with np ∈ 128, 256, 512 pixels. Each image of dimension m×m (m = 1024) was split into 3 / 4 overlapping tiles. Aggressive data augmentation was performed by fragmenting into 9128 tiles to increase redundancy and pattern fragmentation (Figure 3A). For each image M of dimensions m × m, a tile T of size np × np was generated as follows: Each data tile T is rotated by θ ∈ {0°, 90°, 180°, 270°} to represent a different spatial orientation of the ECM (Figure 3A). Thus, from each original image TIFF2024527473000095.tif9128 tiles were saved, resulting in significant data expansion (100x for np = 512, 676x for np = 256, and 3364x for np = 128). The convolutional neural network shown in Fig. 11A was trained on all fragments of the training set for fragment sizes np ∈ 128, 256, 512 and n = 3. The code is provided in Code Sections S1, S2. The learning curves are shown in Fig. 3C and 3D. The most rapid and stable convergence was observed for 512 × 512 pixel imaging, which is not surprising considering the large amount of information per fragment. The trained model was then evaluated to predict annotations for the original images of the validation set. Thus, each original image was TIFF2024527473000096.tif10128 tiles were fragmented into non-overlapping tiles of size np×np (Figure 3B) and the percentage of tiles classified as "hits" was calculated. To determine the accuracy of prediction of the original image independent of the cutoff value of the percentage of tiles classified as "hits", we used receiver operating characteristic analysis (GraphPad Prism v7) and calculated the area under the curve (AUC) for each deep learning model. Different numbers of learning iterations (epochs) and different tile sizes np×np were tested. In general, the accuracy (given as AUC) increased with the number of epochs. After 10,000 epochs, models with np = 256 and np = 128 gave similarly high AUC=0.999 (Figure 3E). Therefore, we selected model 7 with np = 128, n = 3 for further experiments.

[0178] Uniform Manifold Approximation and Projection (UMAP) for Image Clustering Selected image clustering was performed using UMAP (Uniform Manifold Approximation and Projection), a widely used manifold learning method for dimensionality reduction. UMAP is constructed from a theoretical framework based on Riemannian geometry and algebraic topology (35). Each m × m dimensional image pixel matrix (m = 1024) in size is flattened as a linear vector (Figure 3F) and projected into a two-dimensional space using UMAP (Figure 10C). Indeed, in this study, we found that the UMAP approach can generate a 2D data map that logically respects the AI ​​image classification performed in this study, plotting images in two dimensions, which can help us to understand the concept of true positive hits graphically and identify images containing artifacts not recognized by the AI.

[0179] Quantification of αSMA content in primary human lung fibroblasts 6000 cells / well of phLF were seeded in 384-well CellCarrier plates. After overnight culture, cells were starved for 24 h in serum-reduced medium (1% FBS). Then, cells were treated with TGFβ1 (1 ng / ml) and different compounds. After 48 h, cells were fixed with 100% ice-cold methanol. Cells were stained with DAPI and αSMA antibody conjugated to Cy3 (catalog no. C6198-2ML, Sigma). INTEGRA Assist Plus was used for automated liquid handling in 384-well plates. After fixation, automated imaging was achieved using a confocal laser scanning microscope (LSM710, Zeiss) with autofocus detection for three-dimensional image acquisition (ECM deposition assay). Images were analyzed by measuring the mean fluorescence intensity (MFI) of the αSMA signal in Zen Blue v2.5 (Zeiss).

[0180] Contraction assay in primary human lung fibroblasts 50 μl of 3D collagen gel was cast per well in a 96-well imaging plate as previously described (30) and 20.000 phLFs were seeded on top per well. Cells were treated with 1 ng / mL TGFβ1 and / or Example 84. After 72 h, cells were fixed with 4% paraformaldehyde. Collagen gels were imaged using an AxioImager2 (Zeiss) and gel diameters were determined using Zen Blue v2.5 (Zeiss).

[0181] LC-MS / MS of precision-cut lung slices (PCLS) Ten micrograms of each protein extract was digested using a modified FASP protocol (36, 37). Briefly, proteins were reduced and alkylated with dithiothreitol and iodoacetamide, diluted in 4 M urea, and then centrifuged on a 30 kDa filter device (Sartorius). After several washing steps with 8 M urea and 50 mM ammonium bicarbonate, proteins were digested on the filter overnight with Lys-C and trypsin. The resulting peptides were eluted by centrifugation, acidified with TFA, and stored at -20°C. Samples were measured on a QExactive HF-X mass spectrometer (Thermo Scientific) coupled online to an Ultimate 3000 nano-RSLC (Dionex). Tryptic peptides were automatically loaded onto a trapping column (inner diameter (ID) 300 μm × 5 mm, Acclaim PepMap100 C18, 5 μm, 100 Å, LC Packing) followed by C18 reversed-phase chromatography on an analytical column (nanoEase MZ HSS T3 column, 100 Å, 1.8 μm, 75 μm × 250 mm, Waters) at a flow rate of 250 nl / min with a nonlinear acetonitrile gradient from 3% to 40% in 0.1% formic acid over a period of 95 min. Profile precursor spectra from 300 to 1500 m / z were recorded at a resolution of 60000 with an automatic gain control (AGC) target of 3e6 and a maximum injection time of 30 ms. Subsequently, the top 15 fragment spectra of charges 2–7 were recorded at a resolution of 15000 with an AGC target of 1e5, a maximum injection time of 50 ms, an isolation window of 1.6 m / z, a normalized collision energy of 28, and a dynamic exclusion of 30 s. The generated raw files were analyzed with Progenesis QI for proteomics (version 4.1, Nonlinear Dynamics, part of Waters) for label-free quantification as described (38, 39). All MSMS spectra were exported as mgf files using features of charges 2–7. Peptide searches were performed with the Mascot search engine (version 2.6.2) against the Swissprot human protein database (20237 sequences, 11451954 residues).

[0182] Search settings were: precursor tolerance of 10 ppm, fragment tolerance of 0.02 Da, one missed cleavage allowed, carbamidomethylated cysteine ​​as fixed modification, deamidation of glutamine and asparagine allowed as variable modifications, and oxidation of methionine. The peptide false discovery rate (FDR) was 0.46% when applying the percolator algorithm (40). Search results were re-imported into Progenesis QI software. Proteins were quantified by summing the abundance of all unique peptides per protein after normalization to the identified GAPDH and ACTB peptides. The resulting protein abundances were used to calculate fold changes between conditions and repeated measures ANOVA within Progenesis QI software. Proteomic expression data are provided as Table S4.

[0183] Smurf2 siRNA-mediated silencing phLFs were reverse transfected with 2 nM or 10 nM Silencer® pre-designed Smurf2 siRNA (Cat. No.: AM16708, Ambion, ThermoFisher Scientific, Carlsbad, USA) or 10 nM scrambled Silencer® negative control No. 1 siRNA (AM4611, Ambion, ThermoFisher Scientific, Carlsbad, USA) in Lipofectamine® RNAiMax Transfection Reagent (13778-150, ThermoFisher Scientific, Carlsbad, 130 USA) as indicated, and subsequently treated with 1 ng / ml TGFβ1 for 48 h, unless otherwise indicated.

[0184] 3. Evaluation of N23P activity in inhibiting ECM deposition in IPF fibroblasts Experiment Summary:Primary human pulmonary IPF fibroblasts (phLF) derived from three different idiopathic pulmonary fibrosis (IPF) patients (n = 3) were cultured in DMEM F-12 medium supplemented with 20% fetal bovine serum (FBS) and antibiotic supplements. Cells were seeded at 6000 cells / well in 384-well CellCarrier plates. After overnight culture, cells were starved in serum-reduced medium (1% FBS supplemented with 0.1 mM 2-phosphoascorbic acid for 24 hours). phLF were then treated with TGFβ1 (1 ng / ml) or vehicle and cultured with synthetic N23P (10 μM) or appropriate vehicle control. After 72 h of culture, the medium was replaced with starvation medium supplemented with 1 μg / mL AlexaFluor-637 fluorophore-conjugated anti-fibulin 1 antibody (SantaCruz, Clone C-5, Cat. No. sc-25281 AF647) and 1 μg / mL Hoechst H33342 (Sigma). After 4 h of culture, cells were washed three times with PBS and fixed with paraformaldehyde (PFA). After fixation, automated imaging was achieved by using a confocal laser scanning microscope (LSM710, Zeiss) with an automatic focus detection function for three-dimensional image collection (1024px × 1024px × 9px, which corresponds to a dimension of 1417 μm × 1417 μm × 16 μm). For post-acquisition analysis, images were imported into IMARIS software (Bitplane), and the mean fluorescence intensity was quantified and Hoechst-stained cell nuclei were automatically counted by using the spot detection algorithm of Imaris.

[0185] Assessment of ECM Inhibition I:Mean fluorescence intensity (MFI) values ​​represented the extent of fibulin-1 ECM deposition in three different IPF-phLFs (n=3). The inhibitory activity of the tested N23P compounds was graded; we defined the compound as "inactive" if all TGFβ1-induced ECM deposition was preserved (= 0% inhibition by the compound). If no increase in TGFβ1-induced ECM deposition was detected, the compound was assessed as active (100% inhibition by the compound). Based on this definition, three different classes of inhibitory activity were generated: (+++) compounds that showed >_90% inhibition of TGFβ1-induced ECM deposition, (++) compounds that showed 60-90% inhibition, and (+) compounds that showed 20-60% inhibition. Compounds that showed <20% inhibition were classified as inactive. Tranilast shows no inhibition at 10 μM.

[0186] [Table 5.1]

[0187] Assessment of ECM Inhibition II: Mean fluorescence intensity (MFI) values ​​represented the extent of fibulin-1 ECM deposition in one or two different IPF-phLFs (n=1-2). The inhibitory activity of the tested N23P compounds was graded; we defined the compound as "inactive" if all TGFβ1-induced ECM deposition was preserved (= 0% inhibition by the compound). If no increase in TGFβ1-induced ECM deposition was detected, the compound was assessed as active (100% inhibition by the compound). Based on this definition, three different classes of inhibitory activity were generated: (+++) compounds that showed more than 90% inhibition of TGFβ1-induced ECM deposition, (++) compounds that showed 60-90% inhibition, and (+) compounds that showed 20-60% inhibition. Compounds that showed <20% inhibition were classified as inactive.

[0188] [Table 5.2]

[0189] References TIFF2024527473000099.tif224165TIFF2024527473000100.tif250165TIFF2024527473000101.tif226165TIFF2024527473000102.tif236165TIFF2024527473000103.tif52164

Claims

1. A pharmaceutical composition for use in the treatment of fibrosis and / or neoplasms, preferably fibrosis or neoplasms located in the heart, lung, renal tubules, liver, skin, pleura and retroperitoneum, more preferably, the fibrosis is selected from pleural fibrosis, retroperitoneal fibrosis, atrial fibrillation, myocardial interstitial fibrosis, idiopathic pulmonary fibrosis (IPF), interstitial lung disease, chronic kidney disease, non-alcoholic fatty liver disease, skin scar, keloid, tumor-related fibrotic reaction, The pharmaceutical composition comprises a compound according to formula (I): Including the compound according to: Wherein R 1 is -OR 12 , -O(CH 2 ) u (C 3 to C 10 )aryl, -O(CH 2 ) u (C 3 to C 10 )cycloalkyl, -O(CH 2 ) u (C 2 )alkynyl; -(CH 2 ) u (C 3 to C 10 )aryl, -O(CH 2 ) u (C 3 to C 10 )cycloalkyl, -(CH 2 ) u (C 3 to C 10 )cycloalkyl, , and -(CH 2 ) u (C 2 ) alkynyl selected from the group consisting of, u is from 0 to 6; R 2 ~R 5 are independently H, -OR 12 、-(C 1 ~C 10 )alkyl, halogen, cyano, isocyano, cyanato, isocyanato, thiocyanato, isothiocyanato, azido, -(C 2 ~C 10 )alkenyl, -(C 2 ~C 10 )alkynyl, -(C 3 ~C 10 )cycloalkyl, -(C 3 ~C 10 )heterocyclyl, -(C 3 ~C 10 )aryl, -(C 3 ~C 10 )heteroaryl, -CHZ 2 、-CZ 3 -CH 2 Z、-OCHZ 2 、-OCZ 3 、-OCH 2 Z- -N(R 13 )(R 14 )、-N(R 15 )(OR 16 )、-S(O) 0～2 R 17 、-S(O) 1～2 OR 18 、-OS(O) 1～2 R 19 、-OS(O) 1～2 OR 20 、-S(O) 1～2 N(R 21 )(R 22 )、-OS(O) 1～2 N(R 23 )(R 24 )、-N(R 25 )S(O) 1～2 R 26 、-NR 27 S(O) 1～2 OR 28 、-NR 29 S(O) 1～2 N(R 30 )(R 31 )、-C(=X)R 32 、-C(=X)XR 33 , -XC(=X)R 34 , -XC(=X)XR 35 , -OR 36 , -O(CH 2 ) v (C 3 ~C 10 )aryl, -O(CH 2 ) v (C 3 ~C 10 )cycloalkyl, and -O(CH 2 ) v (C 2 )alkynyl selected from the group consisting of; R 6 is selected from the group consisting of H, -(C 1 ~C 10 )alkyl, benzyl and -(CH 2 ) 1～5 (C 3 ~C 10 )cycloalkyl; wherein, -(C 1 ~C 10 )alkyl, benzyl and -(CH 2 ) 1～5 (C 3 ~C 10 )cycloalkyl may be further substituted with at least one substituent selected from the group consisting of halogen, preferably F; R 7 to R 11 are each independently H, -OR 12 -SR 12 -(C 1 to C 10 )alkyl, halogen, -(C 1 to C 10 )alkyloxy(C 1 to C 10 )alkyl, cyano, isocyano, cyanato, isocyanato, thiocyanato, isothiocyanato, azido, -(C 2 to C 10 )alkenyl, -(C 2 to C 10 )alkynyl, -O(C 2 to C 10 )alkynyl, -(C 3 to C 10 )cycloalkyl, -(C 3 to C 10 )heterocyclyl, -(C 3 to C 10 )aryl, -(C 3 to C 10 )heteroaryl, -(CH 2 ) v CHZ 2 -CZ 3 -CH 2 Z, -OCHZ 2 -OCZ 3 -OCH 2 Z, -N(R 13 )(R 14 ), -N(R 15 )(OR 16 ), -S(O) 0～2 R 17 -S(O) 1～2 OR 18 -OS(O) 1～2 R 19 -OS(O) 1～2 OR 20 -S(O) 1～2 N(R 21 )(R 22 ), -OS(O) 1～2 N(R 23 )(R 24 ), -N(R 25 )S(O) 1～2 R 26 , -NR 27 S(O) 1～2 OR 28 , -NR 29 S(O) 1～2 N(R 30 )(R 31 ), -C(=X)R 32 , -C(=X)XR 33 , -XC(=X)R 34 , -XC(=X)XR 35 , -O(CH 2 ) v (C 3 ~C 10 ) cycloalkyl, -O(CH 2 ) v (C 1 ~C 10 ) alkyl and -O(CH 2 ) v (C 3 ~C 10 ) aryl, and is selected from the group consisting of Here, R 1 ~R 5 and R 7 ~R 11 Of the two remaining adjacent portions of and, the formulas (III) to (XI): Optionally form a ring bonded to the basic aromatic ring of formula (I) according to: wherein, T 1 and T 2 are independently selected from the group consisting of H, -(C 1 ~C 10 ) alkyl and halogen; Here, each hydrogen in formulas (III) to (IX) may be halogen, -(C 3 ~C 10 )aryl, or -(C 1 ~C 3 )alkyl, and may preferably be substituted with F; It is selected from O, S, NH, N(C 1 ~C 10 )alkyl; G is selected from CH, N; J 1 to J 4 is independently selected from C or N, preferably J 1 to J 4 is C; Here, J 1 ~J 4 If any one of them is N, each J that is N 1 ~J 4 is not coupled to the corresponding R 1 ~R 4 exists; R 12 to R 36 are independently selected from the group consisting of H, -(C 1 to C 10 ) alkyl, -(C 2 to C 10 ) alkenyl, -(C 2 to C 10 ) alkynyl, -(C 3 to C 10 ) cycloalkyl, -(C 3 to C 10 ) heterocyclyl, -(C 3 to C 10 ) aryl, and -(C 3 to C 10 ) heteroaryl; R 38 is independently selected from the group consisting of H and -(C 1 ~C 10 ) alkyl; Independently -(C 1 ~C 10 )alkyl, -(C 2 ~C 10 )alkenyl, -(C 2 ~C 10 )alkynyl, -(C 3 ~C 10 )cycloalkyl, -(C 3 ~C 10 )heterocyclyl, -(C 3 ~C 10 )aryl, -O(CH 2 ) v (C 3 ~C 10 )cycloalkyl, -O(CH 2 ) v (C 1 ~C 10 )alkyl and -O(CH 2 ) v (C 3 ~C 10 )aryl selected from the group consisting of R 1 ~R 11 , and R 12 ~R 35 is OR 12 , -(C 1 ~C 10 )alkyl, halogen, cyano, isocyano, cyanato, isocyanato, thiocyanato, isothiocyanato, azide, -(C 2 ~C 10 )alkenyl, -(C 2 ~C 10 )alkynyl, -(C 3 ~C 10 )cycloalkyl, -(C 3 ~C 10 )heterocyclyl, -(C 3 ~C 10 )aryl, -(C 3 ~C 10 )heteroaryl, -CHZ 2 , -CZ 3 -CH 2 Z, -OCHZ 2 , -OCZ 3 , -OCH 2 Z-, -N(R 13 )(R 14 )、 -N(R 15 )(OR 16 )、 -NHC(O)(C 1 ~C 10 ) alkyl, -S(O) 0～2 R 17 、 -S(O) 1～2 OR 18 、 -OS(O) 1～2 R 19 、 -OS(O) 1～2 OR 20 、 -S(O) 1～2 N(R 21 )(R 22 )、 -OS(O) 1～2 N(R 23 )(R 24 )、 -N(R 25 )S(O) 1～2 R 26 、 -NR 27 S(O) 1～2 OR 28 、 -NR 29 S(O) 1～2 N(R 30 )(R 31 )、 -C(=X)R 32 、 -C(=X)XR 33 、 -XC(=X)R 34 、 -XC(=X)XR 35 、 -OR 36 、 and -O(CH 2 ) v (C 3 ~C 10 ) aryl, and may be further substituted with at least one substituent selected from the group consisting of; v is from 0 to 5; Z is halogen; X is selected from the group consisting of O, -NH- and S; A is Selected from the group consisting of; n is 1, 2, or 3, preferably 1; o is 1, 2, or 3, preferably 1; R is H, (C 1 - C 6 ) alkyl, cyano, -(C 3 - C 10 ) cycloalkyl, benzyl, or R is by R 7 or R 11 is part of the ring connected thereto, preferably H or benzyl, most preferably H; R 37 is H or -CF 3 ; However, when n is 2 or 3, A may be ; However, R 5 is not -COOH, Pharmaceutical composition.

2. Compound according to formula (II): Wherein R 1 is -OR 12 , -O(C 5 ~C 10 ) heteroaryl, -O(C 3 ~C 10 ) aryl, -O(CH 2 ) u (C 3 ~C 10 ) cycloalkyl, -O(CH 2 ) u (C 3 ~C 10 ) aryl, or ; R 2 to R 5 and R 7 and, and R 11 are independently selected from the group consisting of H, -(C 1 to C 10 )alkyl, halogen, azide, cyano, -O(C 1 to C 10 )alkyl, -(CH 2 ) u (C 3 to C 10 )aryl, -(CH 2 ) u (C 3 to C 10 )cycloalkyl, -(C 2 to C 10 )alkenyl, -(C 2 to C 10 )alkynyl, -(C 3 to C 10 )cycloalkyl, and -(C 3 to C 10 )aryl, and may be further substituted with at least one substituent selected from the group consisting of halogen, -OH, -NH 2 , -NHC(O)CH 3 , -CN, -N 3 , -COOH, and -C(O)NH 2 ; R 6 is H, -(C 1 ~C 10 )alkyl, benzyl or -(CH 2 ) 1～5 (C 3 ~C 10 )cycloalkyl; wherein -(C 1 ~C 10 )alkyl, benzyl and -(CH 2 ) 1～5 (C 3 ~C 10 )cycloalkyl may be further substituted with at least one substituent selected from the group consisting of halogen, preferably F; R 8 and R 9 is H, -O(C 1 to C 10 )alkyl, -SR 12 , -O(CH 2 ) u (C 3 to C 10 )aryl, -O(CH 2 ) u (C 3 to C 10 )cycloalkyl, -O(C 3 to C 10 )cycloalkyl, or -O(C 2 to C 10 )alkenyl; R 10 is H, halogen, -O(C 1 ~C 10 )alkyl, -O(CH 2 ) u (C 3 ~C 10 )aryl, -O(CH 2 ) u (C 3 ~C 10 )cycloalkyl, -O(C 3 ~C 10 )cycloalkyl, or -O(C 2 ~C 10 )alkenyl; However, R 9 when = H, either R 8 or R 10 is -OR 12 ; u is from 0 to 6; R is H, (C 1 ~C 6 )alkyl, cyano, -(C 3 ~C 10 )cycloalkyl, benzyl, or R is by R 7 or R 11 is part of the ring connected thereto, preferably H or benzyl, most preferably H; R 37 is H or -CF 3 ; R 12 is independently H, -(C 1 ~C 10 ) alkyl, -(C 2 ~C 10 ) alkenyl, -(C 2 ~C 10 ) alkynyl, -(C 3 ~C 10 ) cycloalkyl, -(C 3 ~C 10 ) heterocyclyl, -(C 3 ~C 10 ) aryl, -(C 3 ~C 10 ) heteroaryl, (CH 2 ) u (C 3 ~C 10 ) aryl, -(CH 2 ) u (C 3 ~C 10 ) heteroaryl, and -(CH 2 ) u (C 3 ~C 10 ) cycloalkyl; preferably selected from -(C 1 ~C 10 ) alkyl, more preferably from -(C 1 ~C 4 ) alkyl; Here, R 8 ~R 10 Of the two adjacent remaining parts of Optionally form a ring bonded to the basic aromatic ring of formula (II) according to; wherein, T 1 and T 2 are independently selected from the group consisting of H, -(C 1 ~C 10 ) alkyl and halogen; Here, each hydrogen in formulas (III) to (XI) may be halogen, -(C 3 ~C 10 )aryl, or -(C 1 ~C 3 )alkyl, preferably substituted with F; It is O, S, N(C 1 ~C 10 )alkyl or NH; preferably O; R 38 is independently selected from the group consisting of H and -(C 1 ~C 10 ) alkyl; G is selected from CH, N; J 1 ~J 4 is independently selected from C or N, preferably J 1 ~J 4 is C; Here, J 1 ~J 4 If any one of them is N, each J that is N 1 ~J 4 is not coupled to the corresponding R 1 ~R 4 exists; However, J 1 to J 4 when it is C, and I) R 5 is -(CH 2 ) 3 CH 3 ; and R 9 is -OCH 3 , -OCH 2 CH 3 , -O(CH 2 ) 2 CH 3 , -OCH 2 phenyl or -O(CH 2 ) 3 CH 3 ; and R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R are H; and (a)R 8 and R 10 is H; or (b)R 8 is -OCH 3 or -OCH 2 CH 3 and R 10 is H; or (c)R 10 is -OCH 3 or -OCH 2 CH 3 and R 8 is H Case; R 6 is not H; II) R 9 is -OCH 3 ,-O(CH 2 ) 2 CH 3 ,-O(2 - propyl),-O(CH 2 ) 4 CH 3 ,-O(CH 2 ) 5 CH 3 ,-OCH 2 (4 - chlorophenyl),-O(CH 2 ) 2 CH(CH 3 ) 2 ,-OCH 2 (2,6 - dichlorophenyl), or -OCH 2 phenyl; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 , and R are H; R 8 is -OCH 3 and R 10 is H, or R 10 is -OCH 3 and R 8 is H; R 5 is -(CH 2 ) 3 CH 3 is Case; R 6 is not H; III) R 9 is -OCH 3 ; R 8 is Br and R 10 is H, or R 10 is Br and R 8 is H; R 5 is -(CH 2 ) 3 CH 3 ; R 1 , R 2 , R 3 , R 4 , R 11 and R are H In the case where R 1 is not H; IV)R 5 is -(CH 2 ) 3 CH 3 and R 9 is -OCH 3 and R 7 is -OCH 3 and R 11 is H, or R 11 is -OCH 3 and R 7 is H; R 1 , R 2 , R 3 , R 4 , R 8 , R 10 , R 11 and R are H Case; R 6 is not H; V)R 9 is -OCH 3 、-OCH 2 phenyl, or -OCH 2 (2-fluorophenyl); R 8 is Br and R 10 is -OCH 3 or R 10 is Br and R 8 is -OCH 3 ; R 5 is -O(CH 2 ) 3 CH 3 ; R 1 、R 2 、R 3 、R 4 、R 7 、R 11 and R is H In the case, R 6 is not H; VI)R 9 is -O(CH 2 ) 3 CH 3 ; and R 8 is -OCH 2 CH 3 ; and R 10 is H; or R 10 is -OCH 2 CH 3 ; and R 8 is H; R 5 is -O(CH 2 ) 3 CH 3 ; and R 1 , R 2 , R 3 , R 4 , R 7 , R 11 , and R is H In the case where R 6 is not H; VII) R 9 is - OCH 2 (2 - chlorophenyl); R 8 is Br and R 10 is - CH 2 CH 3 or R 10 is Br and R 8 is - OCH 2 CH 3 ; R 5 is -(CH 2 ) 3 CH 3 ; R 1 R 2 R 3 R 4 R 7 R 11 and R is H In the case where R 6 is not H; VIII) R 9 is -O(2-octenyl); R 8 is Cl and R 10 is H, or R 10 is Cl and R 8 is H; R 5 is -(CH 2 ) 3 COOH; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 , and R is H In the case, R6 is not H; IX)R 9 is -OCH 3 ; and R 1 , R 2 , R 3 , R 4 , R 7 , R 8 , R 10 , R 11 , R is H; and R 5 is -(2-fluorophenyl), -phenyl In the case where R 6 is not H; X)R 9 is -OCH 2 CH 3 ; and R 5 is -(CH 2 ) 3 CH 3 ; and R 1 , R 2 , R 3 , R 4 , R 7 , R 8 , R 10 , R 11 , and R is H In the case where R 6 is not H; XI)R 9 is -OCH 3 ; R 8 is -OCH 3 and R 10 is H, or R 10 is -OCH 3 and R 8 is H; R 5 is -(CH 2 ) 3 CH 3 ; R 1 , R 2 , R 3 , R 4 , R 8 , R 7 , and R is H In the case, R 6 is not H; XII)R 9 is -OCH 3 ; R 5 is -O(CH 2 ) 3 CH 3 ; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 , and R is H, and R 8 is -OCH 2 CH 3 and R 10 is H, or R 10 is -OCH 2 CH 3 and R 8 is H In the case, R 6 is not H; XIII)R 5 is -(CH 2 ) 3 CH 3 ; and R 9 is -O(CH 2 ) 3 CH 3 or -OCH 3 ; and R 1 , R 2 , R 3 , R 4 , R 7 , R 8 , R 10 , R 11 , and R is H In the case where R 6 is not H; XIV)R 5 is -CH 3 ,-(CH 2 ) 2 CH 3 or -CH 2 CH 3 ; R 9 is -OCH 3 ; R 8 is -OCH 3 and R 10 is H, or R 8 is H and R 10 is -OCH 3 ; R 1 , R 2 , R 3 , R 4 , R 7 , R 11 and R is H Case; R 6 is not H; XV)R 5 is -(CH 2 ) 3 CH 3 ; and R 1 , R 2 , R 3 , R 4 , R 7 , R 8 , R 9 , R 10 and R 11 are H Case; R 6 is not H.

3. I) R 1 is -OR 12 , -O(CH 2 ) u (C 3 ~C 10 ) aryl, -O(CH 2 ) u (C 3 ~C 10 ) cycloalkyl, and -O(CH 2 ) u (C 2 ) alkynyl; preferably selected from -OR 12 ; u is from 0 to 5; R 12 is -(C 1 ~C 10 )alkyl, preferably -(C 3 ~C 5 )alkyl, more preferably -(C 4 )alkyl, and / or II) A is Selected from, preferably n = 1, 2, more preferably n = 1, and / or III)R 2 ~R 5 is independently H, -OR 12 、-(C 1 ~C 10 )alkyl, halogen, cyano, isocyano, cyanato, isocyanato, thiocyanato, isothiocyanato, azido, -(C 2 ~C 10 )alkenyl, -(C 2 ~C 10 )alkynyl, -(C 3 ~C 10 )cycloalkyl, -(C 3 ~C 10 )heterocyclyl, -(C 3 ~C 10 )aryl, -(C 3 ~C 10 )heteroaryl, -CH(Z) 2 、-C(Z) 3 、-CH 2 Z, -OCH(Z) 2 、-OC(Z) 3 、-OCH 2 Z, -N(R 13 )(R 14 )、-N(R 15 )(OR 16 )、-C(=X)R 32 、-C(=X)XR 33 、-XC(=X)R 34 、-XC(=X)XR 35 、-O(CH 2 ) v (C 3 ~C 10 )aryl, -O(CH 2 ) v (C 3 ~C 10 )cycloalkyl, and -O(CH 2 ) v (C 2 )alkynyl selected from the group consisting of, The pharmaceutical composition according to claim 1.

4. R 7 to R 11 are independently H, -OR 12 , -SR 12 -(C 1 to C 10 ) alkyl, halogen, cyano, isocyano, cyanato, isocyanato, thiocyanato, isothiocyanato, azido, -(C 2 to C 10 ) alkenyl, -(C 2 to C 10 ) alkynyl, -O(C 2 to C 10 ) alkynyl, -(C 3 to C 10 ) cycloalkyl, -(C 3 to C 10 ) heterocyclyl, -(C 3 to C 10 ) aryl, -(C 3 to C 10 ) heteroaryl, -(CH 2 ) v CH(Z) 2 , -C(Z) 3 , -CH 2 Z, -OCH(Z) 2 , -OC(Z) 3 , -OCH 2 Z, -N(R 13 )(R 14 ), -N(R 15 )(OR 16 ), -C(=X)R 32 , -C(=X)XR 33 , -XC(=X)R 34 , -XC(=X)XR 35 , -O(CH 2 ) v (C 3 to C 10 ) cycloalkyl, -O(CH 2 ) v (C 1 to C 10 ) alkyl and -O(CH 2 ) v (C 3 to C 10 ) aryl, and are selected from the group consisting of, Here, R 1 ~R 5 and R 7 ~R 11 Of the remaining two adjacent portions of Optionally form a ring according to, preferably a ring according to formula (III); wherein, T 1 and T 2 are independently selected from the group consisting of H, -(C 1 ~C 10 ) alkyl and halogen; Here, each hydrogen in formulas (III) to (XI) may be substituted with a halogen or -(C 3 ~C 10 )aryl, preferably F; R 38 is independently selected from the group consisting of H and -(C 1 ~C 10 ) alkyl The pharmaceutical composition according to claim 1.

5. R 1 to R 11 is -(C 1 to C 10 )alkyl, -(C 2 to C 10 )alkenyl, -(C 2 to C 10 )alkynyl, -(C 3 to C 10 )cycloalkyl, -(C 3 to C 10 )heterocyclyl, -(C 3 to C 10 )aryl, -O(CH 2 ) v (C 3 to C 10 )cycloalkyl, -O(CH 2 ) v (C 1 to C 10 )alkyl and -O(CH 2 ) v (C 3 to C 10 )aryl and is selected from the group consisting of, and R 12 to R 35 is OR 12 , -(C 1 to C 10 )alkyl, halogen, cyano, isocyano, cyanato, isocyanato, thiocyanato, isothiocyanato, azide, -(C 2 to C 10 )alkenyl, -(C 2 to C 10 )alkynyl, -(C 3 to C 10 )cycloalkyl, -(C 3 to C 10 )heterocyclyl, -(C 3 to C 10 )aryl, -(C 3 to C 10 )heteroaryl, -CHZ 2 , -CZ 3 , -CH 2 Z, -OCHZ 2 , -OCZ 3 , -OCH 2 Z, -N(R 13 )(R 14 ), -N(R 15 )(OR 16 )、 -C(=X)R 32 、 -C(=X)XR 33 、 -XC(=X)R 34 、 and -XC(=X)XR 35 、 -OR 36 、 and -O(CH 2 ) v (C 3 ~C 10 )aryl, and may be further substituted with a substituent selected from the group consisting of: The pharmaceutical composition according to claim 1.

6. R 9 is -OR 12 、-SR 12 、halogen, -O(C 2 ~C 10 ) alkynyl, -CZ 3 、-OCHZ 2 、-OCZ 3 、-OCH 2 Z, -O(CH 2 ) v (C 3 ~C 10 ) cycloalkyl, -O(CH 2 ) v (C 1 ~C 10 ) alkyl and -O(CH 2 ) v (C 3 ~C 10 ) aryl selected from the group consisting of, where -O(C 1 ~C 10 ) alkyl, -OCH 2 Z, -O(CH 2 ) v (C 3 ~C 10 ) cycloalkyl, -O(CH 2 ) v (C 1 ~C 10 ) alkyl and -O(CH 2 ) v (C 3 ~C 10 ) aryl selected from the group consisting of, the pharmaceutical composition according to claim 1, wherein R 9 is optionally further substituted with at least one substituent selected from the group consisting of halogen, -OH, -NH 2 、-NHC(O)CH 3 、-CN、-N 3 、-COOH, and -C(O)NH 2 .

7. (i)R 2 ~R 5 、R 7 or R 11 is independently H, -OR 12 、-(C 1 ~C 10 )alkyl, halogen, cyano, azido, -(C 2 ~C 10 )alkenyl, -(C 2 ~C 10 )alkynyl, -(C 3 ~C 10 )cycloalkyl, -(C 3 ~C 10 )heterocyclyl, -(C 3 ~C 10 )aryl, -(C 3 ~C 10 )heteroaryl, -CHZ 2 、-CZ 3 、-CH 2 Z, -OCHZ 2 、-OCZ 3 、-OCH 2 Z, -OR 36 、-O(CH 2 ) v (C 3 ~C 10 )aryl, -O(CH 2 ) v (C 3 ~C 10 )cycloalkyl, and -O(CH 2 ) v (C 2 )alkynyl, selected from the group consisting of; and / or (ii) R 9 is -OR 12 and R 8 is -H, and R 10 is H or -OR 12 and / or (iii) R 12 is H, -(C 1 ~C 10 ) alkyl, -(C 2 ~C 4 ) alkynyl, -(C 3 ~C 10 ) cycloalkyl, -(C 3 ~C 10 ) heterocyclyl, -(C 3 ~C 10 ) aryl, and -(C 3 ~C 10 ) heteroaryl, selected from the group consisting of The pharmaceutical composition according to claim 1 or the compound according to claim 2.

8. (i)R 9 is H and R 8 is H and R 10 is -OR 12 or; or (ii) R 9 and R 10 are Form a ring bonded to the basic aromatic ring of formula (II) according to, preferably a ring according to formula (III); wherein, T 1 and T 2 are independently selected from the group consisting of H, -(C 1 ~C 10 ) alkyl and halogen; Here, each hydrogen in formulas (III) to (XI) may be substituted with a halogen or -(C 3 ~C 10 )aryl, preferably F; R 38 is independently selected from the group consisting of H, and -(C 1 ~C 10 ) alkyl; Het is O, S or NH, preferably O; G is selected from CH, N; and / or (iii) J 1～4 is CH; and / or (iv) u = 0 to 3; and / or (v)R 12 is -O(C 4 ~C 6 )alkyl, -OCH 2 (C 3 ~C 5 )cycloalkyl, -Ophenyl or -OCH 2 phenyl; and / or (vi)R 6 = H or C 1 ~C 4 alkyl or (CH 2 ) (1～3) alkyl-(C 1 ~C 6 ) cycloalkyl; these may be further substituted with at least one substituent selected from the group consisting of halogen, preferably F, and / or (vii)R 2 ~R 5 、R 7 or R 11 is independently selected from the group consisting of H, -(C 1 ~C 3 )alkyl, halogen, cyano, azido, -CHZ 2 , -CZ 3 , -CH 2 Z, -OCHZ 2 , -OCZ 3 , -OCH 2 Z, and -(C 3 ~C 5 )cycloalkyl; (viii) R 2 is H; and / or (ix)R 7 is H; and / or (x)R 11 is H; and / or (xi) R 3 is H; and / or (xii)R 4 is H; and / or (xiii)R 5 is H and / or (xiv) R 9 is OCH 3 and R 10 is H or R 10 is OCH 3 and / or (xv) R 6 is H; and / or (xvi)R 1 is -O(C -1 ~C 6 ))alkyl; and / or, (xvii)R 10 is H or -OCH 3 ; and / or (xviii) R 1 is -O(C 4 ~C 6 ) alkyl; and / or (xix) R 9 is -OCH 3 and R 10 = H or -OCH 3 ; and / or (xx)R 10 is H, The compound according to claim 2.

9. (i)R 1 is -O(C 1 ~C 4 )alkyl, -O(C 5 ~C 10 )heteroaryl, -O(C 3 ~C 10 )aryl, and -O(CH 2 ) u (C 3 ~C 10 )aryl, selected from the group consisting of, preferably -O butyl; Here, (a) (C 5 ~C 10 ) heteroaryl is preferably Selected from the group consisting of; and / or (b)(C 3 ~C 10 ) The aryl is preferably phenyl and may be substituted with halogen and / or (C 1 ~C 3 ) alkyl; and / or (ii) R 2 is selected from the group consisting of hydrogen and halogen, preferably hydrogen; wherein the halogen is preferably Cl or F; (c)R 3 is selected from the group consisting of hydrogen and halogen, preferably hydrogen; wherein the halogen is preferably Cl or F; and / or (iii)R 4 is selected from the group consisting of hydrogen and halogen, preferably hydrogen; wherein the halogen is preferably Cl or F; and / or (iv) R 5 is selected from the group consisting of hydrogen and halogen, preferably hydrogen; wherein the halogen is preferably Cl or F; and / or (v)R 6 is H, -(C 1 ~C 10 ) alkyl, or -(CH 2 ) 1～5 is cyclopropyl; and / or (vi)R 7 is H; and / or (vii)R 8 is H; and / or (viii)R 9 is H, -O(C 1 ~C 4 )alkyl, -OCH 2 CF 3 , -O(CH 2 )cyclopropyl, -OCF 3 , -OCHF 2 , -O(C 2 ~C 10 )alkenyl, or -O(CH 2 ) 3 is C≡CH; and / or (ix)R 10 is H, halogen, or -O(C 1 ~C 2 ) alkyl; and / or (x) Here, R 9 and the two remaining adjacent parts of R 10 are Form a ring bonded to the basic aromatic ring of formula (II) according to; wherein, T 1 and T 2 are independently selected from the group consisting of H, -methyl and F; Here, each hydrogen in formulas (III) to (XI) may be substituted with methyl; R 38 is independently selected from the group consisting of H, and -(C 1 ~C 10 ) alkyl; Het is O; G is selected from CH; (xi) R 11 is H or -OCH 3 and / or (xii) R is H, C 1 ~C 6 alkyl, or benzyl, most preferably H; and / or (xiii) J 1 ~J 4 is C or N, The compound according to claim 2.

10. A compound selected from the group consisting of, preferably, the compound is Selected from the group consisting of, compound.

11. A pharmaceutical composition for use in the treatment of fibrosis or neoplasms, preferably fibrosis or neoplasms located in the heart, lung, renal tubules, liver, dermis, pleura and retroperitoneum, more preferably, the fibrosis is selected from pleural fibrosis, retroperitoneal fibrosis, atrial fibrillation, myocardial interstitial fibrosis, idiopathic pulmonary fibrosis (IPF), interstitial lung disease, chronic kidney disease, non-alcoholic fatty liver disease, skin scars, keloids, tumor-related fibrotic reactions, The pharmaceutical composition is A pharmaceutical composition comprising a compound selected from the group consisting of

12. A pharmaceutical composition comprising the compound according to claim 2 and at least one carrier.

13. More preferably, the fibrosis is selected from pleural fibrosis, retroperitoneal fibrosis, atrial fibrillation, myocardial interstitial fibrosis, idiopathic pulmonary fibrosis (IPF), interstitial lung disease, chronic kidney disease, non-alcoholic fatty liver disease, skin scars, keloids, tumor-related fibrotic reactions, the pharmaceutical composition according to claim 12 for use in the treatment of fibrosis or neoplasms, preferably fibrosis or neoplasms located in the heart, lung, renal tubules, liver, dermis, pleura and retroperitoneum.

14. The pharmaceutical composition according to claim 1, 11, or 12 for use in the treatment of inflammatory diseases such as articular calcinosis, familial Mediterranean fever and pericarditis.

15. The following steps: (a) Culturing adherent cells that deposit at least one protein in the presence of at least one test compound; (b) Staining at least one protein deposited by the adherent cells; (c) Fixing the adherent cells and the at least one protein; (d) Microscopic detection of the signal of the at least one stained deposited protein; (e) Data analysis of the signal detected in step (d) including quantification of the amount of the at least one protein deposited in the presence of the at least one test compound A screening assay comprising Step (b) is carried out before step (c), Optionally (i) The adherent cells are primary cells and / or (ii) The adherent cells are patient-derived primary human cells, preferably human lung fibroblasts; or animal-derived primary cells; or any adherent immortalized cell line, (iii) Step (a) is carried out for at least 24 hours, preferably 60 - 90 hours, more preferably 65 - 80 hours, most preferably 70 - 75 hours, and / or (iv) Here, at least one protein is an extracellular matrix protein, preferably, at least one extracellular matrix protein is selected from the group consisting of type V collagen, type I collagen, and fibrin 1, and / or (v) Step (b) involves the binding of at least one antibody to at least one protein or to at least one probe bound to at least one protein, where optionally the antibody or probe includes at least one detectable label conjugated directly to the antibody, and where optionally, the detectable label has fluorescence properties, preferably, the detectable label is a fluorophore selected from AlexaFluor 488, AlexaFluor 555, and AlexaFluor 637; AlexaFluor 647, AlexaFluor 568, AlexaFluor 568, and / or Qdot, and / or (vi) Step (b) involves the binding of at least one primary antibody (FA) to at least one protein, and subsequently, the binding of at least one primary antibody (FA) to at least one secondary antibody (SA), where the at least one secondary antibody (SA) includes at least one detectable label conjugated to the antibody, and where optionally, the detectable label has fluorescence properties, preferably, the detectable label is a fluorophore selected from AlexaFluor 488, AlexaFluor 555, and AlexaFluor 637; AlexaFluor 647, AlexaFluor 568, AlexaFluor 568, and / or Qdot, and / or (vii) In step (b), there is at least one additional co-staining and is selected from the group consisting of cell nucleus staining, viability staining, myofibroblast marker (e.g., αSMA staining), apoptosis marker (e.g., caspase 3 / 7 staining), and / or (viii) At least one test compound in step (a) is a small molecule; and / or an oligonucleotide, peptide, protein, proteac, anticalin, antibody, CRISPR, and / or (ix) In step (d), 2D or 3D imaging is performed by a conventional or confocal imaging device, and / or (x) The data analysis in step (e) includes using a machine learning model such as a neural network, a screening assay.

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