Novel anti-fibrotic drugs

A compound according to formula (II)

• wherein,
• R1 is —OR12, —O(C5-C10)heteroaryl, —O(C3-C10)aryl, —O(CH2)u(C3-C1a)cycloalkyl, —O(CH2)u(C3-C10)aryl, or
• R2 to R5 and R7, and R11 are independently selected from the group consisting of H, —(C1-C10)alkyl, halogen, azido, cyano, —O(C1-C10)alkyl, —(CH2)u(C3-C10)aryl, —(CH2)u(C3-C10)cycloalkyl, —(C2-C10)alkenyl, —(C2-C10)alkynyl, —(C3-C10)cycloalkyl, —(C3-C10)aryl, which optionally are 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;
• R6 is H, —(C1-C10)alkyl, benzyl and —(CH2)1-5(C3-C10)cycloalkyl;
• wherein —(C1-C10)alkyl, benzyl and —(CH2)1-5(C3-C10)cycloalkyl optionally are further substituted with at least one substituent selected from the group consisting of Halogen, preferably F;
• R8, R9 and are H, —O(C1-C10)alkyl, —SR12, —O(CH2)u(C3-C1c)aryl, —O(CH2)u(C3-C10)cycloalkyl, —O(C3-C10)cycloalkyl, or, —O(C2-C10)alkenyl;
• R10 is H, halogen, —O(C1-C10)alkyl, —O(CH2)u(C3-C10)aryl, —O(CH2)u(C3-C10)cycloalkyl, —O(C3-C10)cycloalkyl, or, —O(C2-C10)alkenyl;
• with the proviso that if R9=H either Ra or R10 is —OR12
• u is 0 to 6;
• R is H, (C1-C6)alkyl, cyano, —(C3-C10)cycloalkyl, benzyl or part of a ring wherein R is connected with R7 or R11 by
• preferably H or benzyl, most preferably H;
• R37 is H or —CF3;
• R12 are independently selected from the group consisting of H, —(C1-C10)alkyl, —(C2-C10)alkenyl, —(C2-C10)alkynyl, —(C3-C10)cycloalkyl, —(C3-C10)heterocyclyl, —(C3-C10)aryl, —(C3-C10)heteroaryl, (CH2)u(C3-C10)aryl, —(CH2)u(C3-C10)heteroaryl —(CH2)u(C3-C10)cycloalkyl;
• preferably —(C1-C10)alkyl, more preferably —(C1-C4)alkyl;
• wherein two adjacent rests of R8 to R10 optionally may form a ring, attached to the underlying aromatic ring of formula (II) according to
• wherein T1 and T2 are independently selected from the group consisting of H, —(C1-C10)alkyl and halogen;
• wherein each hydrogen in formula (III) to (XI) is optionally substituted with halogen, —(C3-C10)aryl, or —(C1-C3)alkyl, preferably F;
• Het is O, S, N(C1-C10)alkyl or NH;
• preferably O;
• R38 is independently selected from the group consisting of H, —(C1-C10)alkyl;
• G is selected from CH, N;
• J1 to J4 are independently selected from C or N, preferably J1 to J4 are C;
• wherein if any one of J1 to J4 is N, the corresponding R1 to R4 attached to the respective J1 to J4 which is (are) N is absent;
• with the proviso that
• if J1 to J4 are C and
• I) if R5 is —(CH2)3CH3;
• R9 is —OCH3, —OCH2CH3, —O(CH2)2CH3, —OCH2phenyl or —O(CH2)3CH3;
• R1, R2, R, R4, R, R11 and R are H;
• and
• a) R8 and R10 are H;
• or
• b) R8 is —OCH3, or —OCH2CH3 and R10 is H;
• or
• c) R10 is —OCH3, or —OCH2CH3 and R8 is of H;
• then R6 is not H;
• II) if R9 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;
• R1, R2, R, R4, R7, R11, and R are H;
• R8 is —OCH3 and R10 is H, or R10 is —OCH3 and R8 is H;
• R5 is —(CH2)3CH3;
• then R6 is not H;
• III) if R9 is —OCH3;
• R8 is Brand R10 is H, or R10 is Br and R8 is H;
• R5 is —(CH2)3CH3;
• R1, R2, R3, R4, R11 and R are H, then R1 is not H;
• IV) if R5 is —(CH2)3CH3, R9 is —OCH3, R7 is —OCH3 and R11 is H or R11 is —OCH3 and R7 is H;
• R1, R2, R3, R4, R8, R10, R11 and R are H;
• then R6 is not H;
• V) if R9 is —OCH3, —OCH2phenyl, or —OCH2(2-fluorophenyl);
• R8 is Brand R10 is —OCH3, or R10 is Br and R8 is —OCH3;
• R5 is —O(CH2)3CH3;
• R1, R2, R3, R4, R7, R11, and R are H, then R6 is not H;
• VI) if R9 is —O(CH2)3CH3;
• R8 is —OCH2CH3 and R10 is H, or R10 is —OCH2CH3 and R8 is H;
• R5 is —O(CH2)3CH3;
• R1, R2, R3, R4, R7, R11, and R are H, then R6 is not H;
• VII) if R9 is —OCH2(2-chlorophenyl);
• R8 is Br and R10 is —CH2CH3, or R10 is Br and R8 is —OCH2CH3;
• R5 is —(CH2)3CH3;
• R1, R2, R3, R4, R7, R11, and R, are H, then R6 is not H;
• VIII) if R9 is —O(2-octenyl);
• R8 is Cl and R10 is H, or R10 is CI and R8 is H;
• R is —(CH2)3COOH;
• R1, R2, R3, R4, R7, R11, and R are H, then is not H;
• IX) if R9 is —OCH3;
• R1, R2, R3, R4, R7, R8, R10, R11, R are H;
• and
• R5 is -(2-fluorophenyl), -phenyl;
• then R6 is not H;
• X) if R9 is —OCH2CH3;
• R5 is —(CH2)3CH3;
• R1, R2, R3, R4, R7, R8, R10, R11, and R are H;
• then R6 is not H;
• XI) if R9 is —OCH3;
• R8 is —OCH3 and R10 is H, or R10 is —OCH3 and R8 is H;
• R5 is —(CH2)3CH3;
• R1, R2R3, R4, R8, R7, and R are H;
• then R6 is not H;
• XII) if R9 is —OCH3;
• R5 is —O(CH2)3CH3;
• R1, R2, R3, R4, R7, R11, and R are H, and R8 is —OCH2CH3 and R10 is H, or R10 is —OCH2CH3 and R8 is H;
• then R6 is not H;
• XIII) if R5 is —(CH2)3CH3;
• R9 is —O(CH2)3CH3, or —OCH3;
• R1, R2, R3, R4, R7, R8, R10, R11, and R are H;
• then R6 is not H;
• XIV) if R5 is —CH3, —(CH2)2CH3 or —CH2CH3;
• R9 is —OCH3;
• R8 is —OCH3 and R10 is H or R8 is H and R10 is —OCH3;
• R1, R2, R3, R4, R7, R11, and R are H;
• then R6 is not H.
• XV) if R5 is —(CH2)3CH3;
• R1, R2, R3, R4, R7, R8, R9, R10 and R11 are H;
• then R6 is not H.

A screening assay, comprising the steps

• a) culturing adherent cells which deposit at least one protein in the presence of at least one test compound;
• b) staining of at least one protein deposited by the adherent cells;
• c) fixation of adherent cells and the at least one protein;
• d) microscopic detection of a signal of the at least one stained deposited protein;
• e) data analysis of signals detected in step d) comprising quantification of the amount of the at least one protein deposited in the presence of the at least one test compound.
• wherein step b) is carried out before step c)
• wherein optionally
• i) the adherent cells are primary cells and/or
• ii) the adherent cells are primary patient derived human cells, preferably human lung fibroblasts;
• or primary animal derived cells;
• or any adherent immortalized cell lines.
• iii) step a) is carried out for at least 24 h, preferably 60 to 90 hours, more preferably 65 to 80 hours, most preferably for 70 to 75 hours and/or
• iv) wherein at least one protein is an extracellular matrix protein, preferably the at least one extracellular matrix protein is selected from the group consisting of collagen type 5, collagen type 1, and fibulin 1 and/or
• v) step b) comprises the binding of at least one antibody to at least one protein or at least one probe binding to at least one protein and wherein optionally
• the antibody or probe comprises at least one detectable label that is directly conjugated to the antibody and wherein optionally the detectable label has fluorescence property, preferably the detectable lable is a fluorophore selected from AlexaFluor 488, AlexaFluor 555, and AlexaFluor 637;
• AlexaFluor 647, AlexaFluor 568, AlexaFluor 568, and/or Qdots and/or
• vi) step b) comprises the binding of at least one first antibody (FA) to at least one protein and the subsequent binding of 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 that is conjugated to the antibody and wherein optionally the detectable label has fluorescence property, preferably the detectable label is a fluorophore selected from AlexaFluor 488, AlexaFluor 555, and AlexaFluor 637;
• AlexaFluor 647, AlexaFluor 568, AlexaFluor 568, and/or Qdots and/or
• vii) in step b) at least one further co-staining is present and selected from the group consisting of cell-nuclei staining, live-dead staining, myofibroblast markers (e.g. αSMA-staining), apoptosis markers (e.g. Caspase3/7 staining) and/or
• viii) the at least one test compound in step a) is a small molecule;
• and/or oligonucleotides, peptides, proteins, protacs, anticalins, antibodies, CRISPRs and/or
• ix) in step d) 2D or 3D imaging is carried out by a conventional or confocal imaging apparatus and/or
• x) the data analysis in step e) comprises using a machine learning model, such as neural networks.