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SUBSTITUTED PYRIDO[3,4-B]INDOLES FOR THE TREATMENT OF CARTILAGE DISORDERS

20220041596 · 2022-02-10

    Inventors

    Cpc classification

    International classification

    Abstract

    The present invention relates to 8-aryl-substituted and 8-heteroaryl-substituted 9H-pyrido[3,4-b]indoles of the formula (I), in which A, E, G, R.sup.1 to R.sup.6 and R.sup.10 are as defined in the claims, which stimulate chondrogenesis and cartilage matrix synthesis and can be used in the treatment of cartilage disorders and conditions in which a regeneration of damaged cartilage is desired, for example joint diseases such as osteoarthritis. The invention furthermore relates to processes for the synthesis of the compounds of the formula (I), their use as pharmaceuticals, and pharmaceutical compositions comprising them.

    ##STR00001##

    Claims

    1-13. (canceled)

    14: A method of stimulating chondrogenesis or cartilage formation in a subject in need thereof, comprising administering an effective amount of the compound of formula I: ##STR00330## wherein: A is selected from the group consisting of phenyl and a monocyclic or bicyclic, 5-membered to 10-membered, aromatic heterocyclic group, wherein the aromatic heterocyclic group comprises 1 or 2 identical or different hetero ring members selected from the group consisting of N, N(R.sup.20), O and S, and is bonded via a ring carbon atom, and wherein the phenyl and the aromatic heterocyclic group are unsubstituted or substituted on ring carbon atoms by one or more identical or different substituents R.sup.21; E is a direct bond or a chain consisting of 1 to 5 chain members of which 0, 1 or 2 chain members are identical or different hetero chain members selected from the group consisting of N(R.sup.25), O and S(O).sub.m, and the other chain members are identical or different groups C(R.sup.26)(R.sup.27); G is selected from the group consisting of hydrogen, halogen, (C.sub.1-C.sub.4)-alkyl, cyano and R.sup.30; R.sup.1, R.sup.3, R.sup.4 and R.sup.6 are independently selected from the group consisting of hydrogen, halogen and (C.sub.1-C.sub.4)-alkyl; R.sup.2 is selected from the group consisting of hydrogen, halogen, (C.sub.1-C.sub.4)-alkyl and (C.sub.1-C.sub.4)-alkyl-O—C(O)—; R.sup.5 is selected from the group consisting of hydrogen, halogen, (C.sub.1-C.sub.4)-alkyl, (C.sub.1-C.sub.4)-alkyl-O—, cyano, R.sup.7—O—C(O)— and R.sup.8—N(R.sup.9)—C(O)—; R.sup.7, R.sup.1, R.sup.9, R.sup.20, R.sup.22, R.sup.25, R.sup.31, R.sup.33, R.sup.34 and R.sup.40 are independently selected from the group consisting of hydrogen and (C.sub.1-C.sub.4)-alkyl; R.sup.10 is selected from the group consisting of hydrogen, (C.sub.1-C.sub.6)-alkyl, (C.sub.2-C.sub.6)-alkenyl, (C.sub.2-C.sub.6)-alkynyl and (C.sub.3-C.sub.7)-cycloalkyl, wherein the (C.sub.1-C.sub.6)-alkyl group is unsubstituted or substituted by 1 or 2 identical or different substituents selected from the group consisting of (C.sub.3-C.sub.7)-cycloalkyl, Het, cyano and (C.sub.1-C.sub.4)-alkyl-O—, and wherein each (C.sub.3-C.sub.7)-cycloalkyl group is unsubstituted or substituted by one or more identical or different substituents selected from the group consisting of fluorine and (C.sub.1-C.sub.4)-alkyl; R.sup.21 is selected from the group consisting of halogen, (C.sub.1-C.sub.4)-alkyl, (C.sub.1-C.sub.4)-alkyl-O— and cyano, or two groups R.sup.21 attached to adjacent ring carbon atoms in group A are taken together with the carbon atoms to which they are attached to form a 5-membered to 7-membered mono-unsaturated ring comprising 0, 1 or 2 identical or different hetero ring members selected from the group consisting of N(R.sup.22), O and S(O).sub.m, and which is unsubstituted or substituted on ring carbon atoms by one or more identical or different substituents selected from the group consisting of fluorine and (C.sub.1-C.sub.4)-alkyl; R.sup.26 and R.sup.27 are independently selected from the group consisting of hydrogen, fluorine, (C.sub.1-C.sub.4)-alkyl and hydroxy, and in one or two groups C(R.sup.26)(R.sup.27), the substituents R.sup.26 and R.sup.27 attached to the same carbon atom are optionally taken together to form oxo; R.sup.30 is a monocyclic or bicyclic, 3-membered to 10-membered ring, which is saturated or unsaturated and comprises 0, 1, 2 or 3 identical or different hetero ring members selected from the group consisting of N, N(R.sup.31), O and S(O).sub.m, and which is unsubstituted or substituted on ring carbon atoms by one or more identical or different substituents R.sup.32; R.sup.32 is selected from the group consisting of halogen, (C.sub.1-C.sub.4)-alkyl, hydroxy, oxo, (C.sub.1-C.sub.4)-alkyl-O—, cyano, R.sup.33—N(R.sup.34)— and Het; and m is selected from the group consisting of 0, 1 and 2, wherein all numbers m are independent of each other and are identical or different: wherein Het is a monocyclic, 4-membered to 7-membered, saturated heterocyclic group comprising 1 or 2 identical or different hetero ring members selected from the group consisting of N, N(R.sup.40), O and S(O).sub.m, and which is unsubstituted or substituted on ring carbon atoms by one or more identical or different substituents selected from the group consisting of fluorine and (C.sub.1-C.sub.4)-alkyl; and wherein each alkyl group, independently of any other substituents which can be present on an alkyl group, is optionally substituted by one or more fluorine substituents, provided that the compound of formula I is not 8-phenyl-9H-pyrido[3,4-b]indole, or a pharmaceutically acceptable salt thereof, to the subject.

    15: A method of inducing SOX transcription factors in a subject in need thereof, comprising administering an effective amount of the compound of formula I: ##STR00331## wherein: A is selected from the group consisting of phenyl and a monocyclic or bicyclic, 5-membered to 10-membered, aromatic heterocyclic group, wherein the aromatic heterocyclic group comprises 1 or 2 identical or different hetero ring members selected from the group consisting of N, N(R.sup.20), O and S, and is bonded via a ring carbon atom, and wherein the phenyl and the aromatic heterocyclic group are unsubstituted or substituted on ring carbon atoms by one or more identical or different substituents R.sup.21; E is a direct bond or a chain consisting of 1 to 5 chain members of which 0, 1 or 2 chain members are identical or different hetero chain members selected from the group consisting of N(R.sup.25), O and S(O).sub.m, and the other chain members are identical or different groups C(R.sup.26)(R.sup.27); G is selected from the group consisting of hydrogen, halogen, (C.sub.1-C.sub.4)-alkyl, cyano and R.sup.30; R.sup.1, R.sup.3, R.sup.4 and R.sup.6 are independently selected from the group consisting of hydrogen, halogen and (C.sub.1-C.sub.4)-alkyl; R.sup.2 is selected from the group consisting of hydrogen, halogen, (C.sub.1-C.sub.4)-alkyl and (C.sub.1-C.sub.4)-alkyl-O—C(O)—; R.sup.5 is selected from the group consisting of hydrogen, halogen, (C.sub.1-C.sub.4)-alkyl, (C.sub.1-C.sub.4)-alkyl-O—, cyano, R.sup.7—O—C(O)— and R.sup.8—N(R.sup.9)—C(O)—; R.sup.7, R.sup.8, R.sup.9, R.sup.20, R.sup.22, R.sup.25, R.sup.31, R.sup.33, R.sup.34 and R.sup.40 are independently selected from the group consisting of hydrogen and (C.sub.1-C.sub.4)-alkyl; R.sup.10 is selected from the group consisting of hydrogen, (C.sub.1-C.sub.6)-alkyl, (C.sub.2-C.sub.6)-alkenyl, (C.sub.2-C.sub.6)-alkynyl and (C.sub.3-C.sub.7)-cycloalkyl, wherein the (C.sub.1-C.sub.6)-alkyl group is unsubstituted or substituted by 1 or 2 identical or different substituents selected from the group consisting of (C.sub.3-C.sub.7)-cycloalkyl, Het, cyano and (C.sub.1-C.sub.4)-alkyl-O—, and wherein each (C.sub.3-C.sub.7)-cycloalkyl group is unsubstituted or substituted by one or more identical or different substituents selected from the group consisting of fluorine and (C.sub.1-C.sub.4)-alkyl; R.sup.21 is selected from the group consisting of halogen, (C.sub.1-C.sub.4)-alkyl, (C.sub.1-C.sub.4)-alkyl-O— and cyano, or two groups R.sup.21 attached to adjacent ring carbon atoms in group A are taken together with the carbon atoms to which they are attached to form a 5-membered to 7-membered mono-unsaturated ring comprising 0, 1 or 2 identical or different hetero ring members selected from the group consisting of N(R.sup.22), O and S(O).sub.m, and which is unsubstituted or substituted on ring carbon atoms by one or more identical or different substituents selected from the group consisting of fluorine and (C.sub.1-C.sub.4)-alkyl; R.sup.26 and R.sup.27 are independently selected from the group consisting of hydrogen, fluorine, (C.sub.1-C.sub.4)-alkyl and hydroxy, and in one or two groups C(R.sup.26)(R.sup.27), the substituents R.sup.26 and R.sup.27 attached to the same carbon atom are optionally taken together to form oxo; R.sup.30 is a monocyclic or bicyclic, 3-membered to 10-membered ring, which is saturated or unsaturated and comprises 0, 1, 2 or 3 identical or different hetero ring members selected from the group consisting of N, N(R.sup.31), O and S(O).sub.m, and which is unsubstituted or substituted on ring carbon atoms by one or more identical or different substituents R.sup.32; R.sup.32 is selected from the group consisting of halogen, (C.sub.1-C.sub.4)-alkyl, hydroxy, oxo, (C.sub.1-C.sub.4)-alkyl-O—, cyano, R.sup.33—N(R.sup.34)— and Het; and m is selected from the group consisting of 0, 1 and 2, wherein all numbers m are independent of each other and are identical or different; wherein Het is a monocyclic, 4-membered to 7-membered, saturated heterocyclic group comprising 1 or 2 identical or different hetero ring members selected from the group consisting of N, N(R.sup.40), O and S(O).sub.m, and which is unsubstituted or substituted on ring carbon atoms by one or more identical or different substituents selected from the group consisting of fluorine and (C.sub.1-C.sub.4)-alkyl; and wherein each alkyl group, independently of any other substituents which can be present on an alkyl group, is optionally substituted by one or more fluorine substituents, provided that the compound of formula I is not 8-phenyl-9H-pyrido[3,4-b]indole, or a pharmaceutically acceptable salt thereof, to the subject.

    16: A method for treatment of degenerative joint disorders, degenerative cartilage changes, fibrosis, inflammatory processes or pain in a subject in need thereof, comprising administering an effective amount of the compound of formula I: ##STR00332## wherein: A is selected from the group consisting of phenyl and a monocyclic or bicyclic, 5-membered to 10-membered, aromatic heterocyclic group, wherein the aromatic heterocyclic group comprises 1 or 2 identical or different hetero ring members selected from the group consisting of N, N(R.sup.20), O and S, and is bonded via a ring carbon atom, and wherein the phenyl and the aromatic heterocyclic group are unsubstituted or substituted on ring carbon atoms by one or more identical or different substituents R.sup.21; E is a direct bond or a chain consisting of 1 to 5 chain members of which 0, 1 or 2 chain members are identical or different hetero chain members selected from the group consisting of N(R.sup.25), O and S(O).sub.m, and the other chain members are identical or different groups C(R.sup.26)(R.sup.27); G is selected from the group consisting of hydrogen, halogen, (C.sub.1-C.sub.4)-alkyl, cyano and R.sup.30; R.sup.1, R.sup.3, R.sup.4 and R.sup.6 are independently selected from the group consisting of hydrogen, halogen and (C.sub.1-C.sub.4)-alkyl; R.sup.2 is selected from the group consisting of hydrogen, halogen, (C.sub.1-C.sub.4)-alkyl and (C.sub.1-C.sub.4)-alkyl-O—C(O)—; R.sup.5 is selected from the group consisting of hydrogen, halogen, (C.sub.1-C.sub.4)-alkyl, (C.sub.1-C.sub.4)-alkyl-O—, cyano, R.sup.7—O—C(O)— and R.sup.8—N(R.sup.9)—C(O)—; R.sup.7, R.sup.1, R.sup.9, R.sup.20, R.sup.22, R.sup.25, R.sup.31, R.sup.33, R.sup.34 and R.sup.40 are independently selected from the group consisting of hydrogen and (C.sub.1-C.sub.4)-alkyl; R.sup.10 is selected from the group consisting of hydrogen, (C.sub.1-C.sub.6)-alkyl, (C.sub.2-C.sub.6)-alkenyl, (C.sub.2-C.sub.6)-alkynyl and (C.sub.3-C.sub.7)-cycloalkyl, wherein the (C.sub.1-C.sub.6)-alkyl group is unsubstituted or substituted by 1 or 2 identical or different substituents selected from the group consisting of (C.sub.3-C.sub.7)-cycloalkyl, Het, cyano and (C.sub.1-C.sub.4)-alkyl-O—, and wherein each (C.sub.3-C.sub.7)-cycloalkyl group is unsubstituted or substituted by one or more identical or different substituents selected from the group consisting of fluorine and (C.sub.1-C.sub.4)-alkyl; R.sup.21 is selected from the group consisting of halogen, (C.sub.1-C.sub.4)-alkyl, (C.sub.1-C.sub.4)-alkyl-O— and cyano, or two groups R.sup.21 attached to adjacent ring carbon atoms in group A are taken together with the carbon atoms to which they are attached to form a 5-membered to 7-membered mono-unsaturated ring comprising 0, 1 or 2 identical or different hetero ring members selected from the group consisting of N(R.sup.22), O and S(O).sub.m, and which is unsubstituted or substituted on ring carbon atoms by one or more identical or different substituents selected from the group consisting of fluorine and (C.sub.1-C.sub.4)-alkyl; R.sup.26 and R.sup.27 are independently selected from the group consisting of hydrogen, fluorine, (C.sub.1-C.sub.4)-alkyl and hydroxy, and in one or two groups C(R.sup.26)(R.sup.27), the substituents R.sup.26 and R.sup.27 attached to the same carbon atom are optionally taken together to form oxo; R.sup.30 is a monocyclic or bicyclic, 3-membered to 10-membered ring, which is saturated or unsaturated and comprises 0, 1, 2 or 3 identical or different hetero ring members selected from the group consisting of N, N(R.sup.31), O and S(O).sub.m, and which is unsubstituted or substituted on ring carbon atoms by one or more identical or different substituents R.sup.32; R.sup.32 is selected from the group consisting of halogen, (C.sub.1-C.sub.4)-alkyl, hydroxy, oxo, (C.sub.1-C.sub.4)-alkyl-O—, cyano, R.sup.33—N(R.sup.34)— and Het; and m is selected from the group consisting of 0, 1 and 2, wherein all numbers m are independent of each other and are identical or different; wherein Het is a monocyclic, 4-membered to 7-membered, saturated heterocyclic group comprising 1 or 2 identical or different hetero ring members selected from the group consisting of N, N(R.sup.40), O and S(O).sub.m, and which is unsubstituted or substituted on ring carbon atoms by one or more identical or different substituents selected from the group consisting of fluorine and (C.sub.1-C.sub.4)-alkyl; and wherein each alkyl group, independently of any other substituents which can be present on an alkyl group, is optionally substituted by one or more fluorine substituents, provided that the compound of formula I is not 8-phenyl-9H-pyrido[3,4-b]indole, or a pharmaceutically acceptable salt thereof, to the subject.

    Description

    EXAMPLES

    [0247] Abbreviations used are explained below or correspond to the usual conventions. [0248] ACN acetonitrile [0249] BDFP 1,1′-bis(diphenylphosphino)ferrocene-palladium(II) dichloride dichloromethane complex [0250] DCM dichloromethane [0251] DMF N,N-dimethylformamide [0252] DMSO dimethyl sulfoxide [0253] EA ethyl acetate [0254] FA formic acid [0255] HEP n-heptane [0256] MeOH methanol [0257] RT retention time [0258] TFA trifluoroacetic acid [0259] THE tetrahydrofuran

    [0260] When example compounds containing a basic group were purified by preparative high pressure liquid chromatography (HPLC) on reversed phase (RP) column material and, as customary, the eluent was a gradient mixture of water and acetonitrile containing an acid such as trifluoroacetic acid, they were usually obtained in part or completely in the form of their acid addition salts such as the salt with trifluoroacetic acid, depending on the details of the workup such as evaporation or lyophilization conditions. In the names in the heading of the examples and the structural formulae such a trifluoroacetic acid component of an example compound, as well as the acid component of other acid addition salts such as hydrochlorides, for example, in the form of which part of the example compounds have been isolated, is generally not specified.

    [0261] Reactions were generally performed under argon as protective gas. Solvents such as dichloromethane, ethanol, dimethylformamide, methanol, tetrahydrofuran and the like were generally employed as commercially available dry solvents. “Room temperature” means a temperature of 20° C. to 25° C. Reactions under microwave irradiation were carried out in a Personal Chemistry Emrys Optimizer microwave synthesizer in vessels of capacities from 0.5 ml to 20 ml. Solvents were generally evaporated under reduced pressure at temperatures ranging from 35° C. to 45° C. on a rotary evaporator. Chromatography over silica gel was carried out manually (flash chromatography) or supported by semiautomatic cartridge systems such as Companion (CombiFlash) or Flashmaster II (Jones Chromatography). Purifications by preparative RP HPLC were generally performed with columns of a diameter of 25 mm or 30 mm and a length of 250 mm filled with RP18 silica gel of 10 μm particle size, eluting with a gradient of water and acetonitrile containing trifluoroacetic acid or hydrochloric acid.

    [0262] The example compounds were generally characterized by analytical HPLC with ultraviolet detection at 220 nm and 254 nm and mass spectrometry (MS) detection with electrospray ionization (ESI) (LCUV/ESI-MS coupling; LC/MS), and by .sup.1H nuclear magnetic resonance spectroscopy (.sup.1H NMR). The LC/MS analyses were based on the UV chromatograms at 220 nm and 254 nm and the ion current from the mass spectrometer at different ionisation modes (e.g. ESI+, ESI−) with the help of ion extracts of the expected ion masses. .sup.1H NMR spectra were recorded at 400 MHz or 500 MHz or 600 MHz in DMSO-d.sub.6 as solvent at 298 K, unless specified otherwise. In the NMR characterization, the chemical shift 6 (in ppm) and the multiplicities (s=singlet, d=doublet, t=triplet, q=quartet, dd=doublet of doublets, br=broad) and the number of hydrogen atoms (H) of the peaks are given. In the LC/MS characterization, the HPLC method specified below, the retention time (RT) in minutes, and generally the mass-to-charge-ratio m/z of the peak of the molecular ion representing the monoisotopic mass, or of a related ion which was formed depending on the ionization mode, is given. In most cases, the ionization mode was positive electrospray ionization (ESI+), and the mass-to-charge-ratio of the ion [M+H].sup.+ is given. When no significant [M+H].sup.− peak was obtained, the mass-to-charge-ratio of another characteristic mass signal such as [M+2H].sup.++ or an ion of an addition compound with a solvent molecule or [M−H].sup.−, which was formed depending on the ionization mode, such as negative electrospray ionization (ESI−) in the case of the latter ion, is given.

    [0263] The particulars of the HPLC methods in the LC/MS characterization were as follows.

    [0264] Method LC1

    [0265] Column: Merck Chromolith FastGrad RP-18e, 2×50 mm, monolithic; flow: 2.0 ml/min; eluent A: water+0.05% TFA, eluent B: ACN+0.05% TFA; gradient: 98% A: 2% B (0.0 min) to 98% A: 2% B (0.2 min) to 2% A: 98% B (2.4 min) to 2% A: 98% B (3.2 min) to 98% A: 2% B (3.3 min) to 98% A: 2% B (4.0 min)

    [0266] Method LC2

    [0267] Column: Merck Chromolith FastGrad RP-18e, 2×50 mm, monolithic; flow: 2.4 ml/min; eluent A: water+0.05% TFA, eluent B: ACN+0.05% TFA; gradient: 98% A: 2% B (0.0 min) to 98% A: 2% B (0.2 min) to 2% A: 98% B (2.4 min) to 2% A: 98% B (3.2 min) to 98% A: 2% B (3.3 min) to 98% A: 2% B (4.0 min)

    [0268] Method LC3

    [0269] Column: Waters UPLC BEH C18, 2.1×50 mm, 1.7 μm; flow: 0.9 ml/min; temperature 55° C.; eluent A: water+0.05% FA, eluent B: ACN+0.035% FA; gradient: 95% A: 5% B (0.0 min) to 5% A: 95% B (1.1 min) to 5% A: 95% B (1.7 min) to 95% A: 5% B (1.8 min) to 95% A: 5% B (2.0 min)

    [0270] Method LC4

    [0271] Column: Waters UPLC BEH C18, 2.1×50 mm, 1.7 μm; flow: 0.9 ml/min; temperature 55° C.; eluent A: water+0.05% FA, eluent B: ACN+0.035% FA; gradient: 95% A: 5% B (0.0 min) to 5% A: 95% B (2.0 min) to 5% A: 95% B (2.6 min) to 95% A: 5% B (2.7 min) to 95% A: 5% B (3.0 min)

    [0272] Method LC5

    [0273] Column: Waters UPLC BEH C18, 2.1×50 mm, 1.7 μm; flow: 0.9 ml/min; temperature 55° C.; eluent A: water+0.05% FA, eluent B: ACN+0.035% FA; gradient: 98% A: 2% B (0.0 min) to 5% A: 95% B (2.0 min) to 5% A: 95% B (2.6 min) to 98% A: 2% B (2.7 min) to 98% A: 2% B (3.0 min)

    [0274] Method LC6

    [0275] Column: Waters UPLC BEH C18, 2.1×50 mm, 1.7 μm; flow: 0.9 ml/min; temperature 55° C.; eluent A: water+0.1% FA, eluent B: ACN+0.08% FA; gradient: 95% A: 5% B (0.0 min) to 5% A: 95% B (1.1 min) to 5% A: 95% B (1.7 min) to 95% A: 5% B (1.8 min) to 95% A: 5% B (2.0 min)

    [0276] Method LC7

    [0277] Column: Waters XBridge C18, 4.6×50 mm, 2.5 μm; flow: 1.6 ml/min; temperature 30° C.; eluent A: water+0.1% FA, eluent B: ACN+0.08% FA; gradient: 97% A: 3% B (0.0 min) to 2% A: 98% B (18.0 min) to 2% A: 98% B (19.0 min) to 97% A: 3% B (19.5 min) to 97% A: 3% B (20.0 min)

    [0278] Method LC8

    [0279] Column: Waters XBridge C18, 4.6×50 mm, 2.5 μm; flow: 1.3 ml/min; temperature 30° C.; eluent A: water+0.1% FA, eluent B: ACN+0.1% FA; gradient: 97% A: 3% B (0.0 min) to 40% A: 60% B (3.5 min) to 2% A: 98% B (4.0 min) to 2% A: 98% B (5.0 min) to 97% A: 3% B (5.2 min) to 97% A: 3% B (6.5 min)

    [0280] Method LC9

    [0281] Column: Waters XBridge C18, 4.6×50 mm, 2.5 μm; flow: 1.7 ml/min; temperature 50° C.; eluent A: water+0.05% TFA, eluent B: ACN+0.05% TFA; gradient: 95% A: 5% B (0.0 min) to 95% A: 5% B (0.2 min) to 5% A: 95% B (2.4 min) to 5% A: 95% B (3.5 min) to 95% A: 5% B (3.6 min) to 95% A: 5% B (4.5 min)

    [0282] Method LC10

    [0283] Column: Waters XBridge C18, 4.6×50 mm, 2.5 μm; flow: 1.3 ml/min; eluent A: water+0.05% TFA, eluent B: ACN+0.05% TFA; gradient: 95% A: 5% B (0.0 min) to 95% A: 5% B (0.3 min) to 5% A: 95% B (3.5 min) to 5% A:95% B (4.0 min) to 95% A: 5% B (4.5 min)

    [0284] Method LC11

    [0285] Column: YMC-Pack Jsphere H80, 2.1×33 mm, 4.0 μm; flow: 1.0 ml/min; eluent A: water+0.05% TFA, eluent B: ACN+0.05% TFA; gradient: 98% A: 2% B (0.0 min) to 98% A: 2% B (1.0 min) to 5% A: 95% B (5.0 min) to 5% A: 95% B (6.25 min)

    [0286] Method LC12

    [0287] Column: YMC-Pack Jsphere H80, 2.1×33 mm, 4.0 μm; flow: 0.9 ml/min; eluent A: water+0.05% TFA, eluent B: MeOH+0.05% TFA; gradient: 98% A: 2% B (0.0 min) to 98% A: 2% B (1.0 min) to 5% A: 95% B (5.0 min) to 5% A: 95% B (6.25 min)

    [0288] Method LC13

    [0289] Column: Phenomenex Luna C18, 2.0×10 mm, 3.0 μm; flow: 1.1 ml/min; room temperature; eluent A: water+0.05% TFA, eluent B: ACN; gradient: 93% A: 7% B (0.0 min) to 5% A: 95% B (1.2 min) to 5% A: 95% B (1.4 min) to 93% A: 7% B (1.45 min)

    [0290] Method LC14

    [0291] Column: Phenomenex Luna C18, 2.0×10 mm, 3.0 μm; flow: 1.1 ml/min; room temperature; eluent A: water+0.05% TFA, eluent B: ACN; gradient: 93% A: 7% B (0.0 min) to 5% A: 95% B (1.0 min) to 5% A: 95% B (1.45 min) to 93% A: 7% B (1.5 min)

    Exemplary Procedures for the Synthesis of Intermediates

    Intermediate 1. 8-Bromo-6-chloro-9H-pyrido[3,4-b]indole

    [0292] ##STR00011##

    a) 6-Chloro-9H-pyrido[3,4-b]indole

    [0293] N-Chlorosuccinimide (9.78 g, 73.29 mmol) was added in portions with exclusion of light to a solution of norharmane hydrochloride (10.0 g, 48.86 mmol) in water (100 ml) and 1 M hydrochloric acid (100 ml). The mixture was stirred at room temperature overnight and subsequently for 2 h with cooling in ice (0° C. to 5° C.). After dilution with water (50 ml), the precipitate was filtered off with suction, washed with water and dried in a drying cabinet. 7.1 g (76%) of the title compound was obtained.

    [0294] LC/MS (Method LC10): RT=2.26 min; m/z=203.1 [M+H].sup.+

    b) 8-Bromo-6-chloro-9H-pyrido[3,4-b]indole

    [0295] 6-Chloro-9H-pyrido[3,4-b]indole (0.5 g, 2.09 mmol) was placed in water (10 ml) and 1 M hydrochloric acid (10 ml). N-Bromosuccinimide (0.37 g, 2.09 mmol) was added in portions with exclusion of light. The mixture was stirred at room temperature. After 1.5 days conversion to the product was complete, as shown by reaction monitoring by LC/MS. The precipitate was filtered off with suction, washed with water and dried in a drying cabinet to yield 642 mg of the title compound in the form its hydrochloride salt.

    [0296] LC/MS (Method LC10): RT=2.16 min; m/z=281.0 [M+H].sup.+

    Intermediate 2. 8-Bromo-6-chloro-9-cyclopropylmethyl-9H-pyrido[3,4-b]indole

    [0297] ##STR00012##

    a) 6-Chloro-9-cyclopropylmethyl-9H-pyrido[3,4-b]indole

    [0298] 6-Chloro-9H-pyrido[3,4-b]indole (2.0 g, 9.87 mmol) was placed in DMF (40 ml) and treated with cesium carbonate (8.04 g, 24.68 mmol) and cyclopropylmethyl bromide (1.33 g, 0.965 ml, 9.87 mmol). The mixture was stirred at room temperature overnight. The mixture was admixed with water (20 ml) and extracted with EA (3×50 ml). The combined organic phases were washed with a saturated sodium chloride solution, dried over magnesium sulfate and filtered, and the solvent was removed under reduced pressure. 2.5 g (99%) of the title compound was obtained.

    [0299] LC/MS (Method LC6): RT=0.93 min; m/z=257.2 [M+H].sup.+

    b) 8-Bromo-6-chloro-9-cyclopropylmethyl-9H-pyrido[3,4-b]indole

    [0300] 6-Chloro-9-(cyclopropylmethyl)-9H-pyrido[3,4-b]indole (0.5 g, 1.94 mmol) was placed in water (4.78 ml) and 1 M hydrochloric acid (4.78 ml). N-Bromosuccinimide (0.52 g, 2.92 mmol) was added in portions with exclusion of light. The mixture was stirred at room temperature overnight. Further N-bromosuccinimide (0.52 g, 2.92 mmol) was added and the mixture stirred at room temperature for 1 day, when reaction monitoring showed complete conversion to the product. The mixture was extracted with EA (3×20 ml), the combined organic phases were shaken with saturated sodium chloride solution, the organic phase was dried over magnesium sulfate and filtered, and the solvent was removed under reduced pressure. 516 mg of crude product was obtained, which was purified by preparative RP HPLC. The fractions containing the product were combined and concentrated, and the residue freeze-dried to yield 169 mg (19%) of the title compound in the form of its salt with trifluoroacetic acid.

    [0301] LC/MS (Method LC6): RT=1.21 min; m/z=334.9 [M+H].sup.+

    Intermediate 3. 8-Bromo-6-chloro-1-methyl-9H-pyrido[3,4-b]indole

    [0302] ##STR00013##

    a) 6-Chloro-1-methyl-9H-pyrido[3,4-b]indole

    [0303] N-Chlorosuccinimide (1.92 g, 54.32 mmol) was added in portions with exclusion of light to a solution of harmane (2.50 g, 13.72 mmol) in water (60 ml) and 1 M hydrochloric acid (60 ml). The mixture was stirred at room temperature overnight and subsequently for 2 h with cooling in ice (0 to 5° C.). Reaction monitoring by LC/MS showed complete conversion to the product. The precipitate was filtered off with suction, washed with water and dried at 50° C. in a drying cabinet to give 2.2 g (64%) of the title compound in the form of its hydrochloride.

    [0304] LC/MS (Method LC10): RT=2.35 min; m/z=217.0 [M+H].sup.+

    b) 8-Bromo-6-chloro-1-methyl-9H-pyrido[3,4-b]indole

    [0305] 6-Chloro-1-methyl-9H-pyrido[3,4-b]indole hydrochloride (10.00 g, 39.51 mmol) was placed in water (250 ml) and 1 M hydrochloric acid (250 ml). N-Bromosuccinimide (7.03 g, 39.51 mmol) was added in portions with exclusion of light. The mixture was stirred at room temperature overnight. Further N-bromosuccinimide (0.52 g, 2.92 mmol) was added and the mixture stirred at room temperature for 1 day. Reaction monitoring showed complete conversion to the product, and a fine pale yellow precipitate had formed. After cooling for 2 h in ice-water, the precipitate was filtered off with suction and dried to constant weight at 45° C. under reduced pressure to give 13.00 g (99%) of the title compound in the form of its hydrochloride.

    [0306] LC/MS (Method LC11): RT=2.52 min; m/z=295.1 [M+H].sup.+

    Intermediate 4. 8-Bromo-6-chloro-9-cyclopropylmethyl-1-methyl-9H-pyrido[3,4-b]indole

    [0307] ##STR00014##

    [0308] 8-Bromo-6-chloro-1-methyl-9H-pyrido[3,4-b]indole (4.22 g, 12.71 mmol) was placed in DMF (40 ml) and treated with cesium carbonate (10.35 g, 31.78 mmol) and cyclopropylmethyl bromide (1.72 g, 12.71 mmol). The mixture was stirred at room temperature overnight. Reaction monitoring by LC/MS showed complete conversion to the product. The mixture was admixed with water (20 ml) and extracted with EA (3×50 ml). The combined organic phases were washed with saturated sodium chloride solution, dried over magnesium sulfate and filtered, and the solvent was removed under reduced pressure. 4.2 g of crude product was obtained, which was purified by preparative RP HPLC. The fractions containing the product were pooled and concentrated, and the residue freeze-dried to yield 2.22 g (50%) of the title compound in the form of its salt with trifluoroacetic acid.

    [0309] LC/MS (Method LC6): RT=1.13 min; m/z=349.0 [M+H].sup.+

    Intermediate 5. 6-Chloro-8-iodo-1-methyl-9H-pyrido[3,4-b]indole

    [0310] ##STR00015##

    a) 6-Chloro-8-iodo-1-methyl-9H-pyrido[3,4-b]indole

    [0311] 85% phosphoric acid (155 ml) was added to 6-chloro-1-methyl-9H-pyrido[3,4-b]indole hydrochloride (13.1 g). After stirring for 30 min additional phosphoric acid (60 ml) was added. After cooling to 0° C. N-iodosuccinimide (12.8 g) was added in 3 portions within 6 h. After stirring for 16 h in the dark at room temperature further N-iodosuccinimide (3.0 g) was added. After stirring for 24 h the reaction mixture was added to a stirred mixture of ice and water (600 ml). After 30 min the precipitate was filtered off and washed with ice-water. Then 10 N sodium hydroxide solution was added to the filtrate and the pH adjusted to 10. The newly formed precipitate was filtered off with suction and combined with the first precipitate. Water was added to the combined precipitates, and the pH adjusted to 9 with 10 N sodium hydroxide solution. After stirring for 1 h, the solid was filtered off with suction, treated with acetone (250 ml) and filtered off with suction again. This procedure was repeated twice with diethyl ether, and the obtained solid was dried in vacuo at 38° C. Then the solid was dissolved in MeOH with addition of some DMF, and adsorbed to silica gel. After removal of the solvent the silica gel was given on top of a Buchner funnel filled with silica gel. The silica gel was first washed with DCM to remove impurities, and then with a mixture of DCM and MeOH (20:1). The DCM/MeOH filtrate was concentrated in vacuo and the residue was treated with diethyl ether containing some acetone. The solid was filtered off with suction and dried in vacuo to yield 10 g of the title compound.

    [0312] LC/MS (Method LC13): RT=0.74 min; m/z=343.0 [M+H].sup.+

    Intermediate 6. 6-Chloro-9-ethyl-8-iodo-1-methyl-9H-pyrido[3,4-b]indole

    [0313] ##STR00016##

    b) 6-Chloro-9-ethyl-8-iodo-1-methyl-9H-pyrido[3,4-b]indole

    [0314] 6-chloro-8-iodo-1-methyl-9H-pyrido[3,4-b]indole (3.00 g, 8.76 mmol) was dissolved in DMF (25 ml), and cesium carbonate (7.13 g, 21.89 mmol) and iodoethane (858 μl, 10.51 mmol) were added with stirring. After stirring for 16 h under an argon atmosphere, water and DCM were added. After phase separation the aqueous phase was extracted 3 times with DCM. The combined organic layers were washed with brine, dried over sodium sulfate and concentrated in vacuo. The residue was purified by chromatography over silica gel with DCM/MeOH (95:5) to yield 2.1 g of the title compound.

    [0315] LC/MS (Method LC14): RT=0.87 min; m/z=371.1 [M+H].sup.+

    Intermediate 7. 6-Bromo-8-iodo-1-methyl-9H-pyrido[3,4-b]indole

    [0316] ##STR00017##

    a) 6-Bromo-1-methyl-9H-pyrido[3,4-b]indole

    [0317] Harmane (2 g) was suspended in 2 M hydrochloric acid (60 ml) and N-bromosuccinimide (2.15 g) was added with stirring. After stirring for 16 h the reaction mixture was set to pH 9 with 2 N sodium hydroxide solution under cooling. Then EA was added, the phases were separated, and the aqueous phase was extracted 3 times with EA. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography over silica gel with DCM/MeOH (gradient) to yield 1.69 g of the title compound.

    [0318] LC/MS (Method LC4): RT=1.35 min; m/z=261.1 [M+H].sup.+

    b) 6-Bromo-8-iodo-1-methyl-9H-pyrido[3,4-b]indole

    [0319] Phosphoric acid (25 ml) was added to 6-chloro-1-methyl-9H-pyrido[3,4-b]indole (1.56 g), followed by N-iodosuccinimide (1.61 g). The mixture was stirred for 16 h at room temperature in the dark. Then the mixture was adjusted to pH 9 with 10 M sodium hydroxide solution under cooling. EA was added and the phases were separated, and the aqueous phase was extracted 3 times with EA. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography over silica gel with HEP/EA (1:0 to 0:1, gradient) to yield 1.39 g of the title compound.

    [0320] LC/MS (Method LC8): RT=3.05 min; m/z=387.0 [M+H].sup.+

    Intermediate 8. 6-Bromo-9-ethyl-8-iodo-1-methyl-9H-pyrido[3,4-b]indole

    [0321] ##STR00018##

    6-Bromo-9-ethyl-8-iodo-1-methyl-9H-pyrido[3,4-b]indole

    [0322] 6-Bromo-8-iodo-1-methyl-9H-pyrido[3,4-b]indole (700 mg) was dissolved in DMF (10 ml), and cesium carbonate (1.47 g) and iodoethane (180 μl) were added with stirring. After stirring for 2 h under an argon atmosphere additional iodoethane (180 μl) was added and stirring was continued for an additional 2 h. Then water and EA were added. The phases were separated, and the aqueous phase was extracted 3 times with EA. The combined organic phases were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography over silica gel with HEP/EA (gradient)) to yield 650 mg of the title compound.

    [0323] LC/MS (Method LC4): RT=1.65 min; m/z=415.0 [M+H].sup.+

    Intermediate 9. 6-Chloro-8-iodo-1,9-dimethyl-9H-pyrido[3,4-b]indole

    [0324] ##STR00019##

    [0325] 6-Chloro-8-iodo-1-methyl-9H-pyrido[3,4-b]indole (1 g) was dissolved in DMF (10 ml), and cesium carbonate (2.38 g) and iodomethane (220 μl) were added with stirring. After stirring for 16 h under argon atmosphere water and DCM were added. After phase separation, the aqueous phase was extracted 3 times with DCM. The combined organic phases were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography over silica gel with DCM/MeOH (gradient) to yield 440 mg of the title compound.

    [0326] LC/MS (Method LC5): RT=1.38 min; m/z=357.0 [M+H].sup.+

    Intermediate 10. 8-Bromo-6-chloro-9-ethyl-1-methyl-9H-pyrido[3,4-b]indole

    [0327] ##STR00020##

    [0328] The title compound was synthesized from harmane analogously to the synthesis of intermediates 3 and 4 using bromoethane.

    [0329] LC/MS (Method LC12): RT=3.32 min; m/z=323.0 [M+H].sup.+

    Intermediate 11. 8-Bromo-6-chloro-9-ethyl-9H-pyrido[3,4-b]indole

    [0330] ##STR00021##

    [0331] The title compound was synthesized from norharmane analogously to the synthesis of intermediates 3 and 4 using bromoethane.

    [0332] LC/MS (Method LC10): RT=2.73 min; m/z=309.0 [M+H].sup.+

    Intermediate 12. 8-Bromo-6-chloro-1-isopropyl-9H-pyrido[3,4-b]indole

    [0333] ##STR00022##

    a) 1-isopropyl-9H-pyrido[3,4-b]indole

    [0334] A catalytic amount of palladium on charcoal (ca. 100 mg) was added to a solution of 1-isopropyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole (3.00 g, 14 mmol) in xylene (20 ml) and the mixture was stirred at 150° C. for 7 days. The catalyst was separated from the reaction mixture while hot by filtration through a silica gel layer, and the silica gel layer was washed with a small amount of MeOH. The combined organic phases were concentrated and afforded 2.280 g (77%) of the title compound, which was used in the next step without further purification.

    [0335] LC/MS (Method LC6): RT=0.89 min; m/z=209.1 [M−H].sup.−

    b) 6-Chloro-1-isopropyl-9H-pyrido[3,4-b]indole

    [0336] N-Chlorosuccinimide (1.74 g, 12.00 mmol) was added in portions with exclusion of light to a solution of 1-isopropyl-9H-pyrido[3,4-b]indole (2.28 g, 13.72 mmol) in 2 M hydrochloric acid (100 ml). The mixture was stirred at room temperature overnight. Further N-chlorosuccinimide (0.5 g, 3.82 mmol) was added in portions, and the mixture stirred for 1 day. Reaction monitoring by LC/MS showed complete conversion to the product. The mixture was diluted with water (200 ml), neutralized with conc. aqueous sodium hydroxide solution and shaken with EA. The organic phase was separated, dried over magnesium sulfate and concentrated to give 2.60 g (quantitative yield) of the title compound.

    [0337] LC/MS (Method LC6): RT=0.98 min; m/z=245.1 [M+H].sup.+

    c) 8-Bromo-6-chloro-1-isopropyl-9H-pyrido[3,4-b]indole

    [0338] N-Bromosuccinimide (2.73 g, 15.33 mmol) was added in portions with exclusion of light to a solution of 6-chloro-1-isopropyl-9H-pyrido[3,4-b]indole (2.50 g, 10.22 mmol) in 2 M hydrochloric acid (40 ml). The mixture was stirred at room temperature overnight. Reaction monitoring by LC/MS showed complete conversion to the product. The mixture was neutralized with 2 M aqueous sodium hydroxide solution and shaken with EA. The organic phase was separated, dried over magnesium sulfate and concentrated to give 3.30 g (quantitative) of the crude title compound, which was used in the next step without further purification.

    [0339] LC/MS (Method LC6): RT=1.06 min; m/z=323.0 [M+H].sup.+

    Intermediate 13. 8-Bromo-6-chloro-1-ethyl-9H-pyrido[3,4-b]indole

    [0340] ##STR00023##

    [0341] The title compound was synthesized from 1-ethyl-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole analogously to the synthesis of intermediate 12.

    [0342] LC/MS (Method LC6): RT=1.04 min; m/z=309.0 [M+H].sup.+

    Intermediate 14. 8-Bromo-9-but-2-ynyl-6-chloro-9H-pyrido[3,4-b]indole

    [0343] ##STR00024##

    [0344] 8-Bromo-6-chloro-9H-pyrido[3,4-b]indole (2.0 g, 7.1 mmol) in DMF (28 ml) was treated with cesium carbonate (5.79 g, 17.76 mmol) and 1-bromo-but-2-yne (0.95 g, 7.1 mmol). The mixture was stirred at room temperature overnight. Reaction monitoring by LC/MS showed no conversion to the product. Two further additions of the 1-bromo-2-butyne compound, one equivalent each time, were made and the mixture was stirred over several days. The mixture was admixed with water (10 ml) and extracted with EA (3×50 ml). The combined organic phases were washed with saturated sodium chloride solution, dried over magnesium sulfate and filtered, and the solvent was removed under reduced pressure. The product was purified by preparative RP HPLC. The fractions containing the product were pooled and concentrated, and the residue freeze-dried. 809 mg (25%) of the title compound was obtained in the form of its salt with trifluoroacetic acid.

    [0345] LC/MS (Method LC8): RT=3.74 min; m/z=333.1 [M+H].sup.+

    Intermediate 15. 8-Bromo-6-chloro-9-(2,2,2-trifluoroethyl)-9H-pyrido[3,4-b]indole

    [0346] ##STR00025##

    [0347] 8-Bromo-6-chloro-9H-pyrido[3,4-b]indole (5.2 g, 21.75 mmol) in DMF (41 ml) was treated with cesium carbonate (17.72 g, 54.38 mmol) and 2,2,2-trifluoroethyl trifluoromethanesulfonate (5.30 g, 22.84 mmol). The mixture was stirred at room temperature overnight. Reaction monitoring by LC/MS showed complete conversion to the product. The mixture was admixed with water (20 ml) and extracted with EA (3×50 ml). The combined organic phases were washed with saturated sodium chloride solution, dried over magnesium sulfate and filtered, and the solvent was removed under reduced pressure. The crude product was purified by preparative RP HPLC. The fractions containing the product were combined and concentrated, and the residue freeze-dried. 1 g (13%) of the title compound were obtained in the form of its salt with trifluoroacetic acid.

    [0348] LC/MS (Method LC2): RT=1.33 min; m/z=363.0 [M+H].sup.+

    Intermediate 16. 8-Bromo-6-chloro-9-(2-methoxyethyl)-9H-pyrido[3,4-b]indole

    [0349] ##STR00026##

    [0350] 8-Bromo-6-chloro-9H-pyrido[3,4-b]indole (1.0 g, 3.55 mmol) in DMF (10 ml) was treated with cesium carbonate (3.4 g, 17.76 mmol) and 2-bromoethyl methyl ether (0.59 g, 4.26 mmol). The mixture was treated in an ultrasonic bath for 1 h and then stirred at room temperature for 3 days. The solid was filtered off with suction, and the filtrate was concentrated. The crude product was purified by preparative RP HPLC. The fractions containing the product were combined and concentrated, and the residue freeze-dried. 0.56 g (35%) of the title compound was obtained in the form of its salt with trifluoroacetic acid.

    [0351] LC/MS (Method LC8): RT=3.14 min; m/z=339.0 [M+H].sup.+

    Intermediate 17. 6-Chloro-8-iodo-1,5-dimethyl-9H-pyrido[3,4-b]indole

    [0352] ##STR00027##

    a) 1,5-Dimethyl-9H-pyrido[3,4-b]indole

    [0353] Water (150 ml) was added to 4-methyl-DL-tryptophan (1.5 g) at room temperature. Under ice cooling concentrated sulfuric acid (400 μl) and acetaldehyde (585 μl) were added. The mixture was heated to 65° C. for 1.5 h. Then acetic acid (12 ml) was added and the first portion of potassium dichromate (30 mg). After heating to reflux additional 6 portions of potassium dichromate (30 mg) were added until LC/MS control showed complete disappearance of the starting material. After cooling, a saturated sodium carbonate solution was added, followed by solid sodium carbonate to neutralise the solution. Then EA was added, the phases were separated and the aqueous phase was extracted twice with EA. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo to yield 677 mg of the title compound.

    [0354] LC/MS (Method LC5): RT=1.03 min; m/z=197.1 [M+H].sup.+

    b) 6-Chloro-1,5-dimethyl-9H-pyrido[3,4-b]indole

    [0355] 2 N HCl (30 ml) was added to 1,5-dimethyl-9H-pyrido[3,4-b]indole (677 mg) under stirring at room temperature, followed by N-chlorosuccinimide (517 mg). Stirring was continued for 1 h. After standing overnight the pH of the reaction mixture was adjusted to pH 9 by 10 M sodium hydroxide solution under ice cooling. Then EA was added, the phases were separated and the aqueous phase was extracted twice with EA. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography over silica gel with HEP/EA to yield 515 mg of the title compound.

    [0356] LC/MS (Method LC5): RT=1.20 min; m/z=231.1 [M+H].sup.+

    c) 6-Chloro-8-iodo-1,5-dimethyl-9H-pyrido[3,4-b]indole

    [0357] Phosphoric acid (18 ml) was added to 6-chloro-1,5-dimethyl-9H-pyrido[3,4-b]indole (500 mg), followed by N-iodosuccinimide (512 mg), and the mixture stirred for 2.5 h at room temperature in the dark. Then further N-iodosuccinimide (51 mg) was added and stirring was continued for 20 h. The reaction mixture was poured into ice water and the pH adjusted to 9 by 10 M sodium hydroxide solution. The precipitate was filtered off with suction and EA added to the filtrate. The phases were separated and the organic layer dried over sodium sulfate and concentrated in vacuo. The residue was purified by chromatography over silica gel with DCM/ethanol (gradient). The fractions containing the product were combined and concentrated, and the residue freeze-dried to yield 683 mg of the title compound.

    [0358] LC/MS (Method LC5): RT=1.38 min; m/z=356.9 [M+H].sup.+

    Intermediate 18. 8-Bromo-1,6-dimethyl-9H-pyrido[3,4-b]indole

    [0359] ##STR00028##

    a) N-(2-(7-Bromo-5-methyl-1H-indol-3-yl)ethyl)acetamide

    [0360] 2-(7-Bromo-5-methyl-1H-indol-3-yl)ethylamine hydrochloride (3.6 g) was converted into the free base by treatment with 1 M sodium hydroxide solution and DCM, separation of the phases, extraction of the aqueous phase with DCM, and drying of the combined DCM phases over sodium sulfate, filtration and concentration in vacuo. The amine was suspended in dry DCM (60 ml), and triethylamine (2.42 ml) was added. After cooling of the mixture to −40° C., acetyl chloride (1.03 ml) was added with stirring. After 30 min at −30° C. the reaction mixture was poured into ice water (100 ml). The DCM was removed in vacuo, and the remaining aqueous phase was extracted three times with EA. The combined EA phases were dried over sodium sulfate, filtrated and concentrated in vacuo to yield 4.74 g of the crude title compound.

    [0361] LC/MS (Method LC5): RT=1.74 min; m/z=293.2 [M−H].sup.−

    b) 8-Bromo-1,6-dimethyl-4,9-dihydro-3H-pyrido[3,4-b]indole

    [0362] N-(2-(7-Bromo-5-methyl-1H-indol-3-yl)ethyl)acetamide (4.27 g) was dissolved in dry ACN (50 ml), and phosphorus oxychloride (6.62 ml) and phosphorus pentoxide (14.38 g) were added. After heating to 80° C. the reaction mixture was stirred at this temperature for 2 h. Then ice was added and the pH of the mixture was adjusted to 9 with 2 M sodium hydroxide solution. This aqueous mixture was extracted with EA (three times), and the combined EA phases were dried, filtered and concentrated in vacuo. The residue was dissolved in DCM, and the organic phase was extracted with a saturated sodium hydrogencarbonate solution, dried, filtered and concentrated in vacuo to yield 2.1 g of the title compound. The original aqueous phase was additionally extracted with DCM (three times), and the combined DCM phases were dried, filtered and concentrated in vacuo to yield an additional 0.91 g of the title compound.

    [0363] LC/MS (Method LC8): RT=2.81 min; m/z=277.1 [M+H].sup.+

    c) 8-Bromo-1,6-dimethyl-9H-pyrido[3,4-b]indole

    [0364] 8-Bromo-1,6-dimethyl-4,9-dihydro-3H-pyrido[3,4-b]indole (3 g) was suspended in nitrobenzene (25 ml) and heated to 220° C. After 30 min the reaction mixture was cooled to room temperature and purified by chromatography over silica gel, first with HEP, then with DCM/MeOH 9:1. The fractions containing the product were combined and concentrated in vacuo. The residue was subject to a further chromatography over silica gel with DCM/MeOH (gradient). The fractions containing the product combined and concentrated in vacuo to yield 1 g of the title compound.

    [0365] LC/MS (Method LC5): RT=1.44 min; m/z=275.1 [M+H].sup.+

    Intermediate 19. 8-Bromo-9-ethyl-1,6-dimethyl-9H-pyrido[3,4-b]indole

    [0366] ##STR00029##

    [0367] 8-Bromo-1,6-dimethyl-9H-pyrido[3,4-b]indole (1 g) was dissolved in DMF (8 ml), and cesium carbonate (2.96 g) and iodoethane (350 μl) were added with stirring. After stirring for 3 h under an argon atmosphere, water and EA were added. The phases were separated and the aqueous phase was extracted 3 times with EA. The combined organic phases were washed with brine, dried over sodium sulfate and concentrated in vacuo. The residue was purified by chromatography over silica gel with DCM/MeOH to yield: 740 mg of the title compound.

    [0368] LC/MS (Method LC5): RT=1.59 min; m/z=303.2 [M+H].sup.+

    Intermediate 20. 6-Bromo-8-iodo-1,3-dimethyl-9H-pyrido[3,4-b]indole

    [0369] ##STR00030##

    a) 1-(1H-Indol-3-yl)propan-2-one

    [0370] Under argon 2-(1H-indol-3-yl)-N-methoxy-N-methylacetamide (3.00 g, 13.75 mmol) was dissolved in THF (60 ml) and the solution cooled to 0° C. A methylmagnesium bromide solution in THE (27.49 ml, 27.49 mmol) was slowly added with stirring. After 2 h a second portion of methylmagnesium bromide solution (27.49 ml, 27.49 mmol) and after 3 h a third portion of methylmagnesium bromide solution (27.49 ml, 27.49 mmol) were added. Then an aqueous ammonium chloride solution was added, followed by EA. The phases were separated, and the organic phase was washed with water and brine, dried, filtered and concentrated in vacuo. The residue was purified by chromatography over silica gel with HEP/EA (gradient) to yield 2.35 g of the title compound.

    [0371] LC/MS (Method LC5): RT=1.56 min; m/z=174.1 [M+H].sup.+

    b) 1-(2-Acetyl-1H-indol-3-yl)propan-2-one

    [0372] Under an argon atmosphere 1-(1H-indol-3-yl)propan-2-one (2.34 g, 13.5 mmol) was dissolved in diethyl ether (35 ml). The solution was slowly added to zinc chloride (2.76 g, 20.26 mmol) in diethyl ether (50 ml) with stirring at 0° C. Stirring was continued for 30 min and then acetyl chloride (1.92 ml, 27.02 mmol) was added. After stirring for 3 h, ice water was added, followed by an aqueous ammonium chloride solution and EA. The phases were separated, and the organic phase was washed with saturated sodium hydrogencarbonate solution and brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography over silica gel with DCM/MeOH (gradient) to yield 1.39 g of the title compound.

    [0373] LC/MS (Method LC5): RT=1.58 min; m/z=216.1 [M+H].sup.+

    c) 1,3-Dimethyl-9H-pyrido[3,4-b]indole

    [0374] 1-(2-Acetyl-1H-indol-3-yl)propan-2-one (1.38 g, 6.41 mmol) was dissolved in acetic acid (15 ml) and ammonium acetate (988 mg) was added. After stirring for 1 h at 60° C. the reaction mixture was cooled to 0° C. in an ice bath, the pH was set to 9 with 2 M sodium hydroxide solution, and the mixture was extracted three times with DCM. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography over silica gel with DCM/MeOH (gradient) to yield 810 mg of the title compound.

    [0375] LC/MS (Method LC5): RT=1.06 min; m/z=197.1 [M+H].sup.+

    d) 6-Bromo-1,3-dimethyl-9H-pyrido[3,4-b]indole

    [0376] 2 N Hydrochloric acid (35 ml) was added to 1,3-dimethyl-9H-pyrido[3,4-b]indole (812 mg, 4.14 mmol) under stirring at room temperature, followed by N-bromosuccinimide (810 mg, 4.55 mmol). Stirring was continued for 16 h. Then the pH of the reaction mixture was adjusted to pH 9 with 10 M sodium hydroxide solution under ice cooling, EA was added, the phases were separated, and the aqueous phase was extracted twice with EA. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo to yield 1.17 g of the title compound.

    e) 6-Bromo-8-iodo-1,3-dimethyl-9H-pyrido[3,4-b]indole

    [0377] Phosphoric acid (18 ml) was added to 6-bromo-1,3-dimethyl-9H-pyrido[3,4-b]indole (1.14 g, 4.14 mmol), followed by N-iodosuccinimide (1.05 g, 4.56 mmol), and the reaction mixture stirred overnight at room temperature in the dark. Then the mixture was poured into ice water and the pH adjusted to 9 with 10 M sodium hydroxide solution. The precipitate was filtered off with suction, and EA was added to the filtrate. The phases were separated and the aqueous layer was extracted three times with EA. The precipitate was stirred with EA for 15 min, filtered off with suction and washed with further EA. The combined EA phases solutions were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography over silica gel with HEP/EA (gradient) to yield 480 mg of the title compound.

    [0378] LC/MS (Method LC8): RT=3.25 min; m/z=400.9 [M+H].sup.+

    Intermediate 21. 6-Bromo-9-ethyl-8-iodo-1,3-dimethyl-9H-pyrido[3,4-b]indole

    [0379] ##STR00031##

    [0380] 6-Bromo-8-iodo-1,3-dimethyl-9H-pyrido[3,4-b]indole (475 mg, 1.18 mmol) was dissolved in DMF (5 ml), and cesium carbonate (965 mg) and iodoethane (114 μl) were added with stirring. After stirring for 16 h under an argon atmosphere, water and DCM were added. The phases were separated, and the aqueous phase was extracted 3 times with DCM. The combined organic phases were washed with brine, dried over sodium sulfate and concentrated in vacuo. The residue was purified by chromatography over silica gel with DCM/MeOH (gradient) to yield 470 mg of the title compound.

    [0381] LC/MS (Method LC5): RT=1.66 min; m/z=428.9 [M+H].sup.+

    Intermediate 22. 8-Bromo-6-chloro-9-(3-methyloxetan-3-ylmethyl)-9H-pyrido[3,4-b]indole

    [0382] ##STR00032##

    a) 2-Bromo-4-chloro-6-(3-fluoropyridin-4-yl)aniline

    [0383] 2,6-Dibromo-4-chloroaniline (6.5 g) was dissolved in a mixture of DME (180 ml) and water (60 ml). After addition of sodium carbonate (9.66 g) the flask was flushed with argon and the mixture was heated to reflux. 3-Fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (5.13 g) and BDFP (1.86 g) were suspended in dry DMF (40 ml) and added to the reaction mixture via a syringe pump over 5 h. After 2 h further BDFP (0.186 g) was added separately to the reaction mixture. When the addition was finished the mixture was cooled, filtered and concentrated in vacuo, and EA and a saturated sodium hydrogencarbonate solution were added to the residue. The phases were separated, the organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography over silica gel with HEP/EA (1:0 to 2:1). 2.02 g of the title compound was obtained.

    [0384] LC/MS (Method LC5): RT=1.95 min; m/z=301.0 [M+H].sup.+

    b) 2-Bromo-4-chloro-6-(3-fluoropyridin-4-yl)-N-(3-methyloxetan-3-ylmethyl)aniline

    [0385] 2-Bromo-4-chloro-6-(3-fluoropyridin-4-yl)aniline (500 mg) was dissolved in DMF (10 ml), and cesium hydroxide (750 mg) was added. After flushing with argon 3-bromomethyl-3-methyloxetane (330 mg) was added and the reaction mixture was stirred for 64 h at room temperature. Then a saturated sodium hydrogencarbonate solution and EA were added to the mixture. The phases were separated and the aqueous phase was extracted three times with EA. The combined organic phases were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography over silica gel with HEP/EA (gradient) to yield 284 mg of the title compound and 235 mg of 8-bromo-6-chloro-9-(3-methyloxetan-3-ylmethyl)-9H-pyrido[3,4-b]indole (compound of step c)).

    [0386] LC/MS (Method LC5): RT=2.04 min; m/z=385.1 [M+H].sup.+

    c) 8-Bromo-6-chloro-9-(3-methyloxetan-3-ylmethyl)-9H-pyrido[3,4-b]indole

    [0387] 2-Bromo-4-chloro-6-(3-fluoropyridin-4-yl)-N-(3-methyloxetan-3-ylmethyl)aniline (282 mg) was dissolved in THE (20 ml), flushed with argon, and a lithium bis(trimethylsilyl)amide solution (0.73 ml, 0.73 mmol in THF) was added with stirring. After 2 h further lithium bis(trimethylsilyl)amide solution (0.73 ml) was added, and stirring was continued for 16 h. Then a saturated ammonium chloride solution was added, followed by EA, and the phases were separated. The organic phase was washed with a saturated sodium hydrogencarbonate solution and brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography over silica gel with HEP/EA (gradient) to yield 176 mg of the title compound.

    [0388] LC/MS (Method LC5): RT=1.63 min; m/z=365.1 [M+H].sup.+

    Exemplary Procedures for the Synthesis of Compounds of the Formula I

    Example 1. 6-Chloro-9-ethyl-8-pyridin-3-yl-9H-pyrido[3,4-b]indole

    [0389] ##STR00033##

    [0390] Tetrakis(triphenylphosphine)palladium(0) (40.45 mg) was added to a solution of 8-bromo-6-chloro-9-ethyl-pyrido[3,4-b]indole (0.31 g, 1 mmol) in degassed toluene (5 ml) under an argon atmosphere in a 25 ml two-necked flask with reflux condenser. The mixture was stirred for 10 min at room temperature, then treated with a solution of 3-pyridine-boronic acid (147.5 mg, 1.2 mmol) in ethanol and an aqueous sodium carbonate solution (2 M, 0.7 ml), and stirred for 8 h at 100° C. After addition of water (10 ml) the mixture was extracted with EA (3×20 ml). The combined organic layers were washed with brine, dried over potassium sulfate, filtered and concentrated in vacuo. The remaining solid was treated with ACN/TFA (9:1) and an insoluble portion filtered off. The filtrate was concentrated and the residue purified by preparative RP HPLC. The fractions containing the product were combined and lyophilized to yield 65 mg of the title compound in the form of its salt with trifluoroacetic acid.

    [0391] LC/MS (Method LC10): RT=2.15 min; m/z=308.0 [M+H].sup.+

    Example 2. 6-Chloro-8-(1-(pyridin-3-ylmethyl)-1H-pyrazol-4-yl)-9-(2,2,2-trifluoroethyl)-9H-pyrido[3,4-b]indole

    [0392] ##STR00034##

    [0393] Degassed DME (25 ml) and degassed water (8 ml) were charged in a 25 ml microwave reaction flask under argon. 8-Bromo-6-chloro-9-(2,2,2-trifluoroethyl)-9H-pyrido[3,4-b]indole (500 mg, 1.38 mmol), sodium carbonate (583 mg, 5.50 mmol), 1-pyridin-3-ylmethyl-1H-pyrazole-4-boronic acid pinacol ester (588 mg, 2.06 mmol), and BDFP (224 mg, 0.28 mmol) were added, and the mixture was treated in a microwave device at 130° C. for 11 min. The reaction mixture was concentrated and the residue purified by preparative RP HPLC. 510 mg (67%) of the title compound was obtained in the form of its salt with trifluoroacetic acid.

    [0394] LC/MS (Method LC3): RT=1.08 min; m/z=442.0 [M+H].sup.+

    Example 3. 9-(But-2-ynyl)-6-chloro-8-(2,6-dichloropyridin-3-yl)-9H-pyrido[3,4-b]indole

    [0395] ##STR00035##

    [0396] 8-Bromo-9-but-2-ynyl-6-chloro-pyrido[3,4-b]indole (385 mg, 0.86 mmol), cesium carbonate (560 mg, 1.72 mmol), 2,6-dichloropyridin-3-boronic acid pinacol ester (471 mg, 1.72 mmol) and BDFP (201 mg, 0.25 mmol) were reacted and the reaction mixture worked-up analogously as described for the compound of example 47 to yield 84 mg (16%) of the title compound in the form of its salt with trifluoroacetic acid.

    [0397] LC/MS (Method LC12): RT=3.62 min; m/z=400.1 [M+H].sup.+

    Example 4. 2-(6-Chloro-8-(2,6-dichloropyridin-3-yl)-9H-pyrido[3,4-b]indol-9-yl)acetonitrile

    [0398] ##STR00036##

    [0399] 6-Chloro-8-(2,6-dichloropyridin-3-yl)-9H-pyrido[3,4-b]indole (324 mg, 0.7 mmol) was placed in DMF (2 ml) and treated with potassium carbonate (242 mg, 1.75 mmol) and bromoacetonitrile (85 mg, 0.7 mmol). The mixture was stirred at room temperature overnight, then admixed with water (5 ml) and extracted with EA (3×10 ml). The combined organic phases were washed with a saturated sodium chloride solution, dried over magnesium sulfate and filtered, and the solvent was removed under reduced pressure. The crude product was purified by preparative RP HPLC. The fractions containing the product were pooled, concentrated and freeze-dried. 36 mg (10%) of the title compound was obtained in the form of its salt with trifluoroacetic acid.

    [0400] LC/MS (Method LC6): RT=0.99 min; m/z=387.1 [M+H].sup.+

    Example 5. 6-Bromo-9-ethyl-1-methyl-8-(1-pyridin-3-ylmethyl-1H-pyrazol-4-yl)-9H-pyrido[3,4-b]indole

    [0401] ##STR00037##

    [0402] 6-Bromo-9-ethyl-8-iodo-1-methyl-9H-pyrido[3,4-b]indole (200 mg) was dissolved in DME (6 ml) and water (2 ml) in a microwave vessel, and sodium carbonate (204 mg), 3-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)methyl)pyridine (137 mg) and BDFP (79 mg) were added. The mixture was treated for 10 min at 100° C. in a microwave oven. Then further 3-((4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)methyl)pyridine (69 mg) was added and the mixture again treated for 10 min at 100° C. in a microwave oven. After cooling the mixture was filtered and the filtrate concentrated in vacuo. After addition of a saturated sodium hydrogencarbonate solution the mixture was extracted three times with DCM. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. After preparative RP HPLC the fractions containing the product were combined and lyophilized. 145 mg of the title compound were obtained in the form of its salt with trifluoroacetic acid. After addition of a saturated sodium hydrogencarbonate solution to 60 mg of this salt the mixture was extracted three times with DCM. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was treated with water and some ACN added. After lyophilization 48 mg of the title compound were obtained.

    [0403] LC/MS (Method LC5): RT=1.27 min; m/z=446.2 [M+H].sup.+

    Examples 6 and 7. 6-Chloro-9-ethyl-1-methyl-8-[1-(2-methyl-pyridin-3-ylmethyl)-2H-pyrazol-3-yl]-9H-pyrido[3,4-b]indole and 6-chloro-9-ethyl-1-methyl-8-[2-(2-methyl-pyridin-3-ylmethyl)-1H-pyrazol-3-yl]-9H-pyrido[3,4-b]indole

    [0404] 6-Chloro-9-ethyl-1-methyl-8-(1H-pyrazol-3-yl)-9H-pyrido[3,4-b]indole (150 mg) was dissolved in dry DMF (3 ml) in a microwave vessel and cesium carbonate (470 mg) and 3-(bromomethyl)-2-methylpyridine hydrochloride (161 mg) were added with stirring. After treating this mixture for 1 h at 100° C. in a microwave oven the mixture was cooled and further 3-(bromomethyl)-2-methylpyridine hydrochloride (54 mg) was added. After further 1.5 h at 100° C. in the microwave oven the mixture was cooled, filtered and concentrated in vacuo. After addition of a saturated sodium hydrogencarbonate solution the mixture was extracted four times with DCM. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. After preparative RP HPLC the fractions containing each of the two isomeric products were combined and lyophilized.

    Example 6. 6-Chloro-9-ethyl-1-methyl-8-[1-(2-methyl-pyridin-3-ylmethyl)-1H-pyrazol-3-yl]-9H-pyrido[3,4-b]indole

    [0405] ##STR00038##

    [0406] 91 mg of the title compound were obtained in the form of its salt with trifluoroacetic acid, of which 60 mg were treated with sodium hydrogencarbonate as described in example 5 to yield 41 mg of the free base.

    [0407] LC/MS (Method LC4): RT=1.16 min; m/z=416.2 [M+H].sup.+

    Example 7. 6-Chloro-9-ethyl-1-methyl-8-[2-(2-methyl-pyridin-3-ylmethyl)-2H-pyrazol-3-yl]-9H-pyrido[3,4-b]indole

    [0408] ##STR00039##

    [0409] 69 mg of the title compound were obtained in the form of its salt with trifluoroacetic acid, of which 51 mg were treated with sodium hydrogencarbonate as described in example 5 to yield 24 mg of the free base.

    [0410] LC/MS (Method LC4): RT=1.16 min; m/z=416.2 [M+H].sup.+

    Example 8. 6-Chloro-1,5-dimethyl-8-[4-(3-methyl-oxetan-3-ylmethoxy)-phenyl]-9H-pyrido[3,4-b]indole

    [0411] ##STR00040##

    [0412] The title compound was synthesized analogously to the synthesis of the compound of example 27 in two microwave vessels. 170 mg of 6-chloro-8-iodo-1,5-dimethyl-9H-pyrido[3,4-b]indole and 116 mg of 4-(3-methyloxetan-3-ylmethoxy)-phenylboronic acid were used in each run. The reaction mixture was treated for 10 min at 100° C. in a microwave oven. After HPLC purification the fractions containing the product were combined and concentrated to remove the ACN, and then neutralised with a saturated sodium hydrogencarbonate solution. The mixture was extracted twice with EA and the combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was treated with ethanol and water to form a milky suspension which was stirred for 1 h. Then the slurry was concentrated in vacuo and dried in high vacuum overnight. 190 mg of the title compound was obtained.

    [0413] LC/MS (Method LC5): RT=1.56 min; m/z=407.1 [M+H].sup.+

    Example 9. 6-Chloro-8-(2,5-dimethyl-2H-pyrazol-3-yl)-9-ethyl-1-methyl-9H-pyrido[3,4-b]indole

    [0414] ##STR00041##

    [0415] 6-Chloro-9-ethyl-8-iodo-1-methyl-9H-pyrido[3,4-b]indole (320 mg) was dissolved in a mixture of DME (9 ml) and water (3 ml). After addition of sodium carbonate (370 mg) the reaction mixture was flushed with argon. After heating to reflux, 1,3-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (192 mg) and BDFP (70 mg) in dry DMF (3 ml) were added to the reaction mixture via a syringe pump over 4 h. After 1 h an extra portion of 7 mg of BDFP was added to the reaction mixture. When the addition via the syringe pump was finished, the mixture was cooled, filtered and concentrated in vacuo. The crude product was first purified by chromatography over silica gel with DCM/MeOH (gradient) and then by preparative RP HPLC. The fractions containing the product were combined and lyophilized. 187 mg of the title compound were obtained in the form of its salt with trifluoroacetic acid. After addition of a saturated sodium hydrogencarbonate solution to 157 mg of this salt the mixture was extracted three times with DCM. The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo to yield 80 mg of the title compound.

    [0416] LC/MS (Method LC5): RT=1.40 min; m/z=339.2 [M+H].sup.+

    Example 10. 2-(4-[6-Chloro-9-(2,2,2-trifluoroethyl)-9H-pyrido[3,4-b]indol-8-yl]pyrazol-1-yl)ethanol

    [0417] ##STR00042##

    [0418] 2-(4-(6-Chloro-9-(2,2,2-trifluoroethyl)-9H-pyrido[3,4-b]indol-8-yl)-pyrazol-1-yl]-ethyl acetate (238.1 mg, 0.55 mmol) was dissolved in MeOH (4 ml) and 2 equivalents of sodium methoxide (25% solution in MeOH) was added. The reaction mixture was stirred for 2 h at room temperature. The solvent was removed in vacuo and the residue was purified by HPLC. The obtained product was dissolved in EA, washed with a saturated sodium hydrogencarbonate solution, and the organic phase was dried and concentrated in vacuo. The residue was recrystallized from HEP/EA to yield 72.5 mg of the title compound.

    [0419] LC/MS (Method LC4): RT=1.32 min; m/z=395.3 [M+H].sup.+

    Example 11. 6-Chloro-8-(1-phenyl-1H-pyrazol-4-yl)-9-(2,2,2-trifluoroethyl)-9H-pyrido[3,4-b]indole

    [0420] ##STR00043##

    [0421] 6-Chloro-8-(1H-pyrazol-4-yl)-9-(2,2,2-trifluoroethyl)-9H-pyrido[3,4-b]indole (130 mg) was dissolved in dry DMF (10 ml), phenylboronic acid (68 mg), copper(II) acetate (76 mg) and pyridine (66 mg) were added and the resulting mixture was stirred for 2 h. After standing overnight at room temperature the mixture was filtered and concentrated in vacuo. The residue was purified by RP HPLC. The fractions containing the product were combined and lyophilized. 41 mg of the title compound were obtained in the form of its salt with trifluoroacetic acid.

    [0422] LC/MS (Method LC4): RT=1.77 min; m/z=427.2 [M+H].sup.+

    Example 12. tert-Butyl 4-(6-chloro-9-(2,2,2-trifluoroethyl)-9H-pyrido[3,4-b]indol-8-yl)-1H-pyrazole-1-carboxylate

    [0423] ##STR00044##

    [0424] 6-Chloro-8-(1H-pyrazol-4-yl)-9-(2,2,2-trifluoroethyl)-9H-pyrido[3,4-b]indole (94.0 mg, 0.27 mmol) was dissolved in dry DCM (5 ml) and N-ethyl-diisopropylamine (34.64 mg, 0.27 mmol, 50.0 μl), di-tert-butyl dicarbonate (58.49 mg, 0.27 mmol) and 4-dimethylaminopyridine (3.27 mg, 30.0 μmol) were added. The reaction mixture was stirred 2 h at room temperature. Then additional 0.5 equivalents each of N-ethyl-diisopropylamine, di-tert-butyl dicarbonate and 4-dimethylaminopyridine were added, and the reaction mixture was stirred overnight at room temperature. To the reaction mixture water was added, the organic phase was separated, dried, and the solvent was removed in vacuo. The residue was purified by MPLC with HEP/EA. 68.0 mg of the title compound were obtained.

    [0425] LC/MS (Method LC6): RT=1.24 min; m/z=451.0 [M+H].sup.+

    Example 13. 6-Chloro-1-methyl-8-[4-(2-(1H-pyrazol-1-yl)ethoxy)-phenyl]-9H-pyrido[3,4-b]indole

    [0426] ##STR00045##

    [0427] A microwave reaction vessel was charged with 6-chloro-8-iodo-1-methyl-9H-pyrido[3,4-b]indole (250 mg), sodium carbonate (310 mg), 4-[2-(1H-pyrazol-1-yl)ethoxy]-benzeneboronic acid pinacol ester (229 mg), BDFP (119 mg), DME (7.5 ml) and water (2.5 ml). After 10 min at 100° C. in a microwave oven the reaction mixture was filtered and a saturated solution of sodium hydrogencarbonate and DCM were added to the filtrate. After phase separation the aqueous phase was extracted twice with DCM. The combined DCM phases were dried over sodium sulphate, filtered and concentrated in vacuo. The residue was purified by RP HPLC. The fractions containing the product were combined, the ACN was removed in vacuo and the remaining aqueous solution was lyophilized. The residue was treated with a saturated solution of sodium hydrogencarbonate and DCM. After phase separation the aqueous phase was extracted twice with DCM. The combined DCM phases were dried over sodium sulphate, filtered and concentrated in vacuo to yield 207 mg of the title compound.

    [0428] LC/MS (Method LC5): RT=1.63 min; m/z=403.2 [M+H].sup.+

    Example 14. 8-(4-Methoxy-phenyl)-1,6-dimethyl-9H-pyrido[3,4-b]indole

    [0429] ##STR00046##

    [0430] 8-Bromo-1,6-dimethyl-9H-pyrido[3,4-b]indole (200 mg) was dissolved in a mixture of DME (9 ml) and water (3 ml). After addition of sodium carbonate (310 mg) the reaction mixture was flushed with argon. After heating to reflux a mixture of 4-methoxyphenylboronic acid pinacol ester (255 mg) and BDFP (119 mg) in dry DMF (4 ml) were given to the reaction mixture via a syringe pump over 3 h. After 1 h an extra portion of 59 mg of BDFP was added to the reaction mixture. When the addition via the syringe pump was finished, the mixture was cooled, filtered and concentrated in vacuo. The crude product was dissolved in EA, and the EA phase was washed with a saturated sodium hydrogencarbonate solution, dried, filtered and concentrated in vacuo. The residue was purified by a preparative RP HPLC. The fractions containing the product were combined and lyophilized. 30 mg of the title compound were obtained in the form of its salt with trifluoroacetic acid.

    [0431] LC/MS (Method LC5): RT=1.63 min; m/z=303.2 [M+H].sup.+

    Example 15. 9-Ethyl-1,6-dimethyl-8-(1-methyl-1H-pyrazol-4-yl)-9H-pyrido[3,4-b]indole

    [0432] ##STR00047##

    [0433] A microwave reaction vessel was charged with 8-bromo-9-ethyl-1,6-dimethyl-9H-pyrido[3,4-b]indole (200 mg), sodium carbonate (280 mg), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (206 mg), BDFP (108 mg), DME (9 ml) and water (3 ml). After 12 min at 130° C. in a microwave oven the mixture was filtered and the filtrate concentrated in vacuo. The residue was dissolved in EA. The resulting solution was washed with a saturated sodium hydrogencarbonate solution, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing the product were combined and lyophilized. 7 mg of the title compound were obtained in the form of its salt with trifluoroacetic acid.

    [0434] LC/MS (Method LC5): RT=1.43 min; m/z=305.2 [M+H].sup.+

    Example 16. 6-Bromo-1,3-dimethyl-8-[4-(2-pyrazol-1-yl-ethoxy)-phenyl]-9H-pyrido[3,4-b]indole

    [0435] ##STR00048##

    [0436] A microwave reaction vessel was charged with 6-bromo-8-iodo-1,3-dimethyl-9H-pyrido[3,4-b]indole (200 mg), 1-(2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)ethyl)-1H-pyrazole (157 mg), BDFP (81 mg), sodium carbonate (211 mg), DME (6 ml) and water (2 ml). After 10 min at 100° C. in a microwave oven the mixture was cooled and a saturated sodium hydrogencarbonate solution followed by DCM was added. The phases were separated and the organic phase was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing the product were combined and lyophilized. 162 mg of the title compound was obtained in the form of its salt with trifluoroacetic acid.

    [0437] LC/MS (Method LC5): RT=1.67 min; m/z=461.2 [M+H].sup.+

    Example 17. 6-(6-Chloro-9-ethyl-1-methyl-9H-pyrido[3,4-b]indol-8-yl)-3-methoxy-pyridin-2-ylamine

    [0438] ##STR00049##

    [0439] A microwave reaction vessel was charged with 6-chloro-9-ethyl-8-iodo-1-methyl-9H-pyrido[3,4-b]indole (250 mg), 6-amino-5-methoxypyridin-2-ylboronic acid (860 mg), BDFP (111 mg), sodium carbonate (71 mg), DME (6 ml) and water (2 ml). After 10 min at 100° C. in a microwave oven the mixture was cooled and filtered. 1 N hydrochloric acid was added to the filtrate, which was washed twice with DCM. The aqueous phase was set to pH 9 with 1 N sodium hydroxide solution and extracted three times with DCM. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing the product were combined, the ACN was removed in vacuo and the aqueous residue lyophilized to yield 50 mg of the title compound in the form of its salt with trifluoroacetic acid.

    [0440] LC/MS (Method LC5): RT=1.42 min; m/z=367.2 [M+H].sup.+

    Example 18. 6-Chloro-9-ethyl-1-methyl-8-(3-phenyl-isoxazol-5-yl)-9H-pyrido[3,4-b]indole

    [0441] ##STR00050##

    [0442] A microwave reaction vessel was charged with 6-chloro-9-ethyl-8-iodo-1-methyl-9H-pyrido[3,4-b]indole (200 mg), 3-phenyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole (176 mg), BDFP (89 mg), sodium carbonate (286 mg), DME (8 ml) and water (3 ml). After 15 min at 100° C. in a microwave oven the mixture was cooled, filtered, and a saturated sodium hydrogencarbonate solution followed by DCM were added to the filtrate. The phases were separated and the aqueous phase was extracted three times with DCM. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing the product were combined, the ACN was removed in vacuo and the remaining aqueous solution was lyophilized to yield 178 mg of the title compound in the form of its salt with trifluoroacetic acid. 135 mg of this salt was treated with a saturated sodium hydrogencarbonate solution and DCM. The phases were separated and the aqueous phase was extracted twice with DCM. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was treated with water, filtered off with suction and dried in high vacuum at 40° C. to yield 106 mg of the title compound.

    [0443] LC/MS (Method LC8): RT=3.79 min; m/z=388.1 [M+H].sup.+

    Examples 19 and 20. 6-Chloro-9-ethyl-1-methyl-8-(2-methyl-2H-pyrazol-3-yl)-9H-pyrido[3,4-b]indole and 6-chloro-9-ethyl-1-methyl-8-(1-methyl-1H-pyrazol-3-yl)-9H-pyrido[3,4-b]indole

    [0444] 6-Chloro-9-ethyl-1-methyl-8-(1H-pyrazol-3-yl)-9H-pyrido[3,4-b]indole (95 mg) was dissolved in dry DMF (3 ml), and sodium hydride (15 mg) was added with stirring. After stirring for 30 min, iodomethane (48 mg) was added and stirring was continued for an additional 16 h. Then EA was added and the solution was washed with water and brine. The organic phase was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing each of the two isomeric products were combined, the ACN was removed in vacuo, the aqueous residues were set basic with a saturated hydrogencarbonate solution and extracted three times with DCM. The combined organic phases were dried over sodium sulfate, filtered, concentrated in vacuo, and the residue dissolved in water/ACN and lyophilized.

    Example 19. 6-Chloro-9-ethyl-1-methyl-8-(2-methyl-2H-pyrazol-3-yl)-9H-pyrido[3,4-b]indole

    [0445] ##STR00051##

    [0446] 20 mg of the title compound, the more polar of the two isomers, were obtained.

    [0447] LC/MS (Method LC4): RT=1.30 min; m/z=325.2 [M+H].sup.+

    Example 20. 6-Chloro-9-ethyl-1-methyl-8-(1-methyl-1H-pyrazol-3-yl)-9H-pyrido[3,4-b]indole

    [0448] ##STR00052##

    [0449] 32 mg of the title compound, the less polar of the two isomers, was obtained.

    [0450] LC/MS (Method LC4): RT=1.32 min; m/z=325.2 [M+H].sup.+

    Example 21. 6-Bromo-9-ethyl-1,3-dimethyl-8-(1-methyl-1H-pyrazol-4-yl)-9H-pyrido[3,4-b]indole

    [0451] ##STR00053##

    [0452] In a microwave reaction vessel (20 ml) 6-bromo-9-ethyl-8-iodo-1,3-dimethyl-9H-pyrido[3,4-b]indole (465 mg, 1.08 mmol) was dissolved in a mixture of DME (12 ml) and water (4 ml). Then 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (225.48 mg, 1.08 mmol), sodium carbonate (459.43 mg, 4.33 mmol) and BDFP (177 mg) were added and the mixture was treated for 10 min at 100° C. in a microwave oven. Then further 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (112 mg) and BDFP (89 mg) were added and the mixture was heated for 12 min at 120° C., followed by additional 15 min at 130° C., in a microwave oven. After cooling a saturated sodium hydrogencarbonate solution and DCM were added and the phases were separated. The organic phase was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing the product were combined and lyophilized to yield 280 mg of the title compound in the form of its salt with trifluoroacetic acid.

    [0453] LC/MS (Method LC5): RT=1.52 min; m/z=383.1 [M+H].sup.+

    Example 22. 6-Chloro-8-(4-methoxy-phenyl)-1,9-dimethyl-9H-pyrido[3,4-b]indole

    [0454] ##STR00054##

    [0455] In a microwave reaction vessel (10 ml) 6-chloro-8-iodo-1,9-dimethyl-9H-pyrido[3,4-b]indole (200 mg, 561 μmol) was dissolved in a mixture of DME (6 ml) and water (2 ml). Then 2-(4-methoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (131 mg, 561 μmol), sodium carbonate (238 mg, 2.24 mmol) and BDFP (92 mg, 110 μmol) were added, and the mixture was treated for 10 min at 100° C. and then for 15 min at 120° C. in a microwave oven. After cooling, a sodium hydrogencarbonate solution and DCM were added and the phases were separated. The aqueous phase was extracted three times with DCM. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing the product were combined and lyophilized. 47 mg of the title compound in the form of its salt with trifluoroacetic acid was obtained.

    [0456] LC/MS (Method LC4): RT=1.52 min; m/z=337.2 [M+H].sup.+

    Example 23. 6-Chloro-1-methyl-8-(2-methyl-2,3-dihydro-benzofuran-5-yl)-9H-pyrido[3,4-b]indole

    [0457] ##STR00055##

    [0458] In a microwave reaction vessel (10 ml) 6-chloro-8-iodo-1-methyl-9H-pyrido[3,4-b]indole (200 mg, 580 μmol) was dissolved in a mixture of DME (6 ml) and water (2 ml). Then 2-methyl-2,3-dihydrobenzofuran-5-ylboronic acid (103.92 mg, 583.83 μmol), sodium carbonate (248 mg) and BDFP (95 mg) were added and the mixture was treated for 10 min at 100° C. in a microwave oven. After cooling, a saturated sodium hydrogencarbonate solution and DCM were added and the phases were separated. The aqueous phase was extracted three times with DCM. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing the product were combined and lyophilized to yield 120 mg of the title compound in the form of its salt with trifluoroacetic acid. 88 mg of this salt were treated with a saturated sodium hydrogencarbonate solution and DCM. The aqueous phase was removed by means of a Chem Elut cartridge, and the organic phase was concentrated in vacuo to yield 52 mg of the title compound.

    [0459] LC/MS (Method LC4): RT=1.56 min; m/z=349.2 [M+H].sup.+

    Example 24. [4-(6-Chloro-1-methyl-9H-pyrido[3,4-b]indol-8-yl)-phenyl]-phenyl-methanol

    [0460] ##STR00056##

    [0461] Under an argon atmosphere 4-(6-chloro-1-methyl-9H-pyrido[3,4-b]indol-8-yl)-benzaldehyde (60 mg) was dissolved in in dry THF with stirring. The solution was cooled to 0 00 and a phenyl magnesium bromide solution (0.41 ml; 1 M in THF) was added with stirring. After the addition was complete the ice bath was removed. After 20 h water was added, and the aqueous phase was extracted three times with EA. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography over silica gel with DCM/MeOH (gradient). The fractions containing the product were combined and concentrated in vacuo to yield 38 mg of the title compound.

    [0462] LC/MS (Method LC4): RT=1.54 min; m/z=399.2 [M+H].sup.+

    Example 25. 6-Chloro-1,9-dimethyl-8-(1-methyl-1H-pyrazol-4-yl)-9H-pyrido[3,4-b]indole

    [0463] ##STR00057##

    [0464] In a microwave reaction vessel (10 ml) 6-chloro-8-iodo-1,9-dimethyl-9H-pyri do[3,4-b]indole (206 mg, 577 μmol) was dissolved in a mixture of DME (6 ml) and water (2 ml). Then 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (120 mg, 577.70 μmol), sodium carbonate (244.92 mg, 2.31 mmol) and BDFP 94 mg were added and the mixture was treated for 15 min at 120° C. in a microwave oven. To complete the conversion, the mixture was treated for another 15 min at 120° C. in a microwave oven. After cooling, a sodium hydrogencarbonate solution and DCM were added and the phases were separated. The aqueous phase was extracted three times with DCM. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing the product were combined and lyophilized. The obtained product was further purified by another preparative RP HPLC, followed by chromatography over silica gel with DCM/MeOH (gradient), to yield 12 mg of the title compound.

    [0465] LC/MS (Method LC4): RT=1.24 min; m/z=311.2 [M+H].sup.+

    Example 26. (2-[4-(6-Chloro-9-ethyl-1-methyl-9H-pyrido[3,4-b]indol-8-yl)-phenoxy]-ethyl)-diisopropyl-amine

    [0466] ##STR00058##

    a) 8-(4-(2-Bromoethoxy)phenyl)-6-chloro-9-ethyl-1-methyl-9H-pyrido[3,4-b]indole

    [0467] ##STR00059##

    [0468] The title compound was synthesized analogously to the synthesis of the compound of example 31, using 4-(2-bromoethoxy)phenylboronic acid.

    [0469] LC/MS (Method LC8): RT=3.89 min; m/z=443.2 [M+H].sup.+

    b) (2-[4-(6-Chloro-9-ethyl-1-methyl-9H-pyrido[3,4-b]indol-8-yl)-phenoxy]-ethyl)-diisopropyl-amine

    [0470] In a microwave vessel 8-(4-(2-bromoethoxy)phenyl)-6-chloro-9-ethyl-1-methyl-9H-pyrido[3,4-b]indole (70 mg) was dissolved in diisopropylamine (3 ml). The mixture was treated for 10 h at 100° C. in a microwave oven. After cooling the amine was removed in vacuo and the residue was purified by chromatography over silica gel with DCM/MeOH (gradient). The fractions containing the product were combined and concentrated in vacuo. The residue was dissolved in a mixture of ACN and water containing 0.05% hydrogen chloride, and the solution lyophilized. 14 mg of the title compound was obtained in the form of (2-[4-(6-chloro-9-ethyl-1-methyl-9H-pyrido[3,4-b]indol-8-yl)-phenoxy]-ethyl)-diisopropyl-amine dihydrochloride.

    [0471] LC/MS (Method LC8): RT=2.74 min; m/z=464.3 [M+H].sup.+

    Example 27. 6-Chloro-8-(4-methoxy-phenyl)-1,5-dimethyl-9H-pyrido[3,4-b]indole

    [0472] ##STR00060##

    [0473] In a microwave reaction vessel (10 ml) 6-chloro-8-iodo-1,5-dimethyl-9H-pyri do[3,4-b]indole (180 mg, 504.78 μmol) was dissolved in a mixture of DME (6 ml) and water (2 ml). Then 2-(4-methoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (118.17 mg, 504.78 μmol), sodium carbonate (214.00 mg, 2.02 mmol) and BDFP (82.44 mg, 100.96 μmol) were added, and the mixture was treated for 15 min at 120° C. in a microwave oven. After cooling, a sodium hydrogencarbonate solution and DCM were added and the phases were separated. The aqueous phase was extracted three times with DCM. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing the product were combined and lyophilised. 92 mg of the title compound in the form of its salt with trifluoroacetic acid was obtained.

    [0474] LC/MS (Method LC4): RT=1.56 min; m/z=337.2 [M+H].sup.+

    Example 28. 6-Chloro-9-ethyl-1-methyl-8-[4-(2-morpholin-4-yl-ethoxy)-phenyl]-9H-pyrido[3,4-b]indole

    [0475] ##STR00061##

    [0476] In a microwave vessel 8-(4-(2-bromoethoxy)phenyl)-6-chloro-9-ethyl-1-methyl-9H-pyrido[3,4-b]indole (31 mg) was dissolved in morpholine (2 ml). The mixture was treated for 15 min at 100° C. in a microwave oven. After cooling the amine was removed in vacuo and the residue was purified by chromatography over silica gel with DCM/MeOH (gradient). The fractions containing the product were combined and concentrated in vacuo. The residue was dissolved in a mixture of ACN and water containing 0.05% hydrochloric acid and lyophilized. 25 mg of the title compound in the form of 6-chloro-9-ethyl-1-methyl-8-[4-(2-morpholin-4-yl-ethoxy)-phenyl]-9H-pyrido[3,4-b]indole dihydrochloride was obtained.

    [0477] LC/MS (Method LC8): RT=2.48 min; m/z=450.3 [M+H].sup.+

    Example 29. [4-(6-Chloro-1-methyl-9H-pyrido[3,4-b]indol-8-yl)-benzyl]-cyclopentyl-amine

    [0478] ##STR00062##

    [0479] 4-(6-Chloro-1-methyl-9H-pyrido[3,4-b]indol-8-yl)-benzylamine (80 mg) was dissolved in DME (2 ml) and acetic acid (0.250 ml). After the addition of cyclopentanone (127 mg) the reaction mixture war stirred for 15 min at room temperature. Then sodium triacetoxyborohydride (111 mg) was added. After stirring for 2 h, DCM was added to the reaction mixture and the solution washed with a saturated sodium hydrogencarbonate solution and brine. The organic phase was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing the product were combined, the ACN was removed in vacuo and the residue was lyophilized to yield 88 mg of the title compound in the form of its salt with trifluoroacetic acid. 65 mg of this salt was treated a with saturated sodium hydrogencarbonate solution and EA. The phases were separated, and the aqueous phase was extracted twice with EA. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo to yield 44 mg of the title compound.

    [0480] LC/MS (Method LC4): RT=1.15 min; m/z=390.2 [M+H].sup.+

    Example 30. 6-Chloro-9-ethyl-1-methyl-8-(4-morpholin-4-ylmethyl-phenyl)-9H-pyrido[3,4-b]indole

    [0481] ##STR00063##

    [0482] 4-(6-Chloro-9-ethyl-1-methyl-9H-pyrido[3,4-b]indol-8-yl)benzaldehyde (94 mg) was dissolved in DME (2 ml) and acetic acid (0.16 ml). After addition of morpholine (28 mg) the reaction mixture was stirred for 15 min at room temperature. Then sodium triacetoxyborohydride (144 mg) was added. After stirring for 16 h, DCM was added to the reaction mixture, the solution washed with a saturated sodium hydrogencarbonate solution and brine. The organic phase was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing the product were combined, the ACN was removed in vacuo and the residue was lyophilised to yield 45 mg of the title compound in the form of its salt with trifluoroacetic acid.

    [0483] LC/MS (Method LC4): RT=1.07 min; m/z=420.3 [M+H].sup.+

    Example 31. 4-(6-Chloro-1-methyl-9H-pyrido[3,4-b]indol-8-yl)-benzaldehyde

    [0484] ##STR00064##

    [0485] In a microwave reaction vessel (10 ml) 6-chloro-8-iodo-1-methyl-9H-pyrido[3,4-b]indole (150 mg) was dissolved in a mixture of DME (5 ml) and water (1.5 ml). Then 4-formylphenylboronic acid (66 mg), sodium carbonate (185 mg) and BDFP (72 mg) were added and the mixture was treated for 10 min at 100° C. in a microwave oven. After cooling, water and DCM were added and the phases were separated. The aqueous phase was extracted three times with DCM. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing the product were combined, the ACN was removed in vacuo and the residue was neutralised with a saturated sodium hydrogencarbonate solution. After extraction with DCM (three times) the organic phases were combined, dried over sodium sulfate, filtered and concentrated in vacuo to yield 60 mg of the title compound. A part of this product was dissolved in a mixture of water, ACN and TFA and lyophilised to yield 10 mg of the title compound in the form of its salt with trifluoroacetic acid.

    [0486] LC/MS (Method LC4): RT=1.40 min; m/z=321.1 [M+H].sup.+

    Example 32. [4-(6-Chloro-1-methyl-9H-pyrido[3,4-b]indol-8-yl)-phenyl]-pyridin-2-yl-methanol

    [0487] ##STR00065##

    [0488] Under an argon atmosphere 4-(6-chloro-9-ethyl-1-methyl-9H-pyrido[3,4-b]indol-8-yl)benzaldehyde (60 mg) was dissolved in dry THE (10 ml) with stirring. The solution was cooled to 0° C. and 2-pyridylmagnesium bromide (2.24 ml; 0.25 M in THF) was added with stirring. After the addition was complete, the ice bath was removed. After 1 h, further 2-pyridylmagnesium bromide (1.12 ml) was added. After 1.5 h a saturated ammonium chloride solution was added, and the aqueous phase was extracted three times with EA. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography over silica gel with HEP/EA (gradient). The fractions containing the product were combined and concentrated in vacuo. The product was purified by preparative RP HPLC. The fractions containing the product were combined, the ACN removed in vacuo, and the residue was lyophilized to yield 60 mg of the title compound in the form of its salt with trifluoroacetic acid.

    [0489] LC/MS (Method LC4): RT=1.30 min; m/z=400.2 [M+H].sup.+

    Example 33. 8-(4-Methoxy-phenyl)-1-methyl-9H-pyrido[3,4-b]indole-6-carbonitrile

    [0490] ##STR00066##

    [0491] In a microwave vessel 6-bromo-8-(4-methoxyphenyl)-1-methyl-9H-pyrido[3,4-b]indole (100 mg, 272.30 μmol) was dissolved in NMP (5 ml) and copper(I) cyanide (487.77 mg, 5.45 mmol) was added. The mixture was treated for 2 h at 200° C. in a microwave oven. After cooling, a saturated ammonium chloride solution was added and the aqueous phase was extracted three times with EA. The combined organic phases were dried with sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing the product were combined, the ACN was removed in vacuo and the residue was lyophilised to yield 30 mg of the title compound in the form of its salt with trifluoroacetic acid.

    [0492] LC/MS (Method LC4): RT=1.32 min; m/z=314.2 [M+H].sup.+

    Example 34. 6-Chloro-1-methyl-8-[4-(quinolin-2-ylmethoxy)-phenyl]-9H-pyrido[3,4-b]indole

    [0493] ##STR00067##

    a) 4-(6-Chloro-1-methyl-9H-pyrido[3,4-b]indol-8-yl)phenol

    [0494] ##STR00068##

    [0495] A microwave reaction vessel was charged with 6-chloro-8-iodo-1-methyl-9H-pyrido[3,4-b]indole (200 mg), 4-hydroxyphenylboronic acid (201 mg), BDFP (96 mg), sodium carbonate (248 mg), DME (8 ml) and water (3 ml). After 15 min at 100° C. in a microwave oven the mixture was cooled, filtered and concentrated in vacuo. EA was added to the residue, and the organic phase was washed twice with water, dried over sodium sulfate, filtered and concentrated in vacuo to yield 167 mg of the title compound, which was directly used in the next step.

    b) 6-Chloro-1-methyl-8-[4-(quinolin-2-ylmethoxy)-phenyl]-9H-pyrido[3,4-b]indole

    [0496] 4-(6-Chloro-1-methyl-9H-pyrido[3,4-b]indol-8-yl)phenol (167 mg) was dissolved in DMF (5 ml). After addition of 2-(chloromethyl)quinoline hydrochloride (125 mg) the reaction mixture war stirred for 3 h at 60° C. Then water was added and the aqueous phase was extracted three times with EA. The combined organic phases were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing the product were combined, the ACN was removed in vacuo, and the precipitate formed was filtered off with suction and dried at 40° C. to yield 50 mg of the title compound in the form of its salt with trifluoroacetic acid. 36 mg of this salt was treated with a saturated sodium hydrogencarbonate solution, the solid was filtered off with suction, washed with water and dried at 40° C. to yield 20 mg of the title compound.

    [0497] LC/MS (Method LC4): RT=1.62 min; m/z=450.3 [M+H].sup.+

    Examples 35 and 36. 8-(4-Methoxy-phenyl)-1-methyl-9H-pyrido[3,4-b]indole-6-carboxylic acid and 8-(4-Methoxy-phenyl)-1-methyl-9H-pyrido[3,4-b]indole-6-carboxamide

    [0498] 8-(4-Methoxy-phenyl)-1-methyl-9H-pyrido[3,4-b]indole-6-carbonitrile trifluoroacetic acid salt (21 mg) was suspended in concentrated sulfuric acid (3 ml) under cooling in an ice bath, and heated for 4 h at 90° C. After cooling, the mixture was concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing each of the two products were combined, the ACN was removed in vacuo, and the residue was lyophilised.

    Example 35. 8-(4-Methoxy-phenyl)-1-methyl-9H-pyrido[3,4-b]indole-6-carboxylic acid

    [0499] ##STR00069##

    [0500] 4 mg of the title compound were obtained in the form of its salt with trifluoroacetic acid.

    [0501] LC/MS (Method LC4): RT=1.15 min; m/z=333.2 [M+H])

    Example 36. 8-(4-Methoxy-phenyl)-1-methyl-9H-pyrido[3,4-b]indole-6-carboxamide

    [0502] ##STR00070##

    [0503] 7 mg of the title compound were obtained in the form of its salt with trifluoroacetic acid.

    [0504] LC/MS (Method LC8): RT=2.33 min; m/z=332.2 [M+H].sup.+

    Example 37. 8-[3-(5-Bromo-pyrimidin-2-yloxy)-phenyl]-6-chloro-1-methyl-9H-pyrido[3,4-b]indole

    [0505] ##STR00071##

    a) 3-(6-Chloro-1-methyl-9H-pyrido[3,4-b]indol-8-yl)phenol

    [0506] ##STR00072##

    [0507] Three microwave reaction vessels were each charged with 6-chloro-8-iodo-1-methyl-9H-pyrido[3,4-b]indole (200 mg), 3-hydroxyphenylboronic acid (200 mg), sodium carbonate (247 mg), BDFP (96 mg), DME (8 ml) and water (3 ml). After 15 min at 100° C. in a microwave oven the mixtures of the three vessels were combined, filtered and concentrated in vacuo. EA was added to the residue and the organic phase was washed twice with water, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was treated with diethyl ether and filtered off with suction to yield 541 mg of the title compound, which was directly used in the next step.

    b) 8-[3-(5-Bromo-pyrimidin-2-yloxy)-phenyl]-6-chloro-1-methyl-9H-pyrido[3,4-b]indole

    [0508] 4-(6-Chloro-1-methyl-9H-pyrido[3,4-b]indol-8-yl)phenol (135 mg) was dissolved in DMF (5 ml). After addition of 5-bromo-2-chloropyrimidine (101 mg), the reaction mixture war stirred for 3 h at 60° C. Then water was added and the aqueous phase was extracted three times with DCM. The combined organic phases were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fraction containing the product were combined, the ACN was removed in vacuo and the residue was lyophilized. The product was further purified by chromatography over silica gel with EA/HEP (1:1 to 1:0). The fractions containing the product were combined and concentrated in vacuo. The residue was lyophilized in water/TFA to yield 48 mg of the title compound in the form of its salt with trifluoroacetic acid.

    [0509] LC/MS (Method LC5): RT=1.54 min; m/z=465.1 [M+H].sup.+

    Example 38. 6-Bromo-9-ethyl-1-methyl-8-(1-quinolin-2-ylmethyl-1H-pyrazol-4-yl)-9H-pyrido[3,4-b]indole

    [0510] ##STR00073##

    [0511] In a microwave reaction vessel (10 ml) dry DMF (5 ml) was added to 6-bromo-9-ethyl-1-methyl-8-(1H-pyrazol-4-yl)-9H-pyrido[3,4-b]indole (50 mg, 140.75 μmol) and 2-(chloromethyl)quinoline hydrochloride (45.20 mg, 211.13 μmol), followed by cesium carbonate (138 mg) The mixture was treated for 2 h at 120° C. in a microwave oven. Then further cesium carbonate (69 mg) was added and the mixture treated for another 2 h at 120° C. in a microwave oven. After filtration, the mixture was concentrated in vacuo, and the residue was purified by preparative RP HPLC. The fractions containing the product were combined, the ACN was removed in vacuo and the residue lyophilised to yield 42 mg of the title compound in the form of its salt with trifluoroacetic acid.

    [0512] LC/MS (Method LC4): RT=1.50 min; m/z=496.3 [M+H].sup.+

    Example 39. 6-Chloro-1-methyl-8-[4-(1-methyl-1H-imidazol-2-ylmethoxy)-phenyl]-9H-pyrido[3,4-b]indole

    [0513] ##STR00074##

    [0514] Dry DMF (4 ml) was added to 4-(6-chloro-1-methyl-9H-pyrido[3,4-b]indol-8-yl)phenol (90 mg), 2-(chloromethyl)-1-methyl-1H-imidazole hydrochloride (49 mg) and potassium carbonate (201 mg), and the mixture was stirred for 2 h at 60° C. Then water was added, and the precipitate was filtered off with suction and purified by chromatography over silica gel with EA/HEP (2:3 to 1:0), followed by DCM. The fractions containing the product were combined and concentrated in vacuo. The residue was dissolved in 1 N hydrochloric acid and the solution washed with DCM. Then a 1 N sodium hydroxide solution was added and the precipitate was filtered off. After washing with water the precipitate was dried at 40° C. to yield 23 mg of the title compound.

    [0515] LC/MS (Method LC5): RT=1.11 min; m/z=403.2 [M+H].sup.+

    Example 40. 6-Chloro-8-[4-([1,4]dioxan-2-ylmethoxy)-phenyl]-1-methyl-9H-pyrido[3,4-b]indole

    [0516] ##STR00075##

    [0517] Dry DMF (4 ml) was added to 4-(6-chloro-1-methyl-9H-pyrido[3,4-b]indol-8-yl)phenol (100 mg), 1,4-dioxan-2-ylmethyl 4-methylbenzenesulfonate (97 mg) and cesium carbonate (530 mg), and the mixture was stirred for 4 h at 80° C. Then a saturated sodium hydrogencarbonate solution was added, and the aqueous phase was extracted three times with DCM. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing the product were combined, the ACN was removed in vacuo and the residue was lyophilized to yield 62 mg of the title compound in the form of its salt with trifluoroacetic acid. 33 mg of this salt was treated with a saturated sodium hydrogencarbonate solution and DCM. After phase separation the aqueous phase was extracted twice with DCM. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo to yield 12 mg of the title compound.

    [0518] LC/MS (Method LC5): RT=1.45 min; m/z=409.3 [M+H].sup.+

    Example 41. 6-Chloro-1-methyl-8-[4-(quinazolin-2-ylmethoxy)-phenyl]-9H-pyrido[3,4-b]indole

    [0519] ##STR00076##

    [0520] Dry DMF (4 ml) was added to 4-(6-chloro-1-methyl-9H-pyrido[3,4-b]indol-8-yl)phenol (100 mg), 2-(chloromethyl)quinazoline (61 mg) and potassium carbonate (224 mg), and the mixture was stirred for 4 h at 80° C. Then further potassium carbonate (10 mg) was added and the mixture stirred for another 3 h at 80° C. After cooling, a saturated sodium hydrogencarbonate solution was added and the aqueous phase was extracted three times with DCM. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fractions containing the product were combined, the ACN was removed in vacuo and the residue was lyophilized to yield 37 mg of the title compound in the form of its salt with trifluoroacetic acid. 29 mg of this salt was treated with a saturated sodium hydrogencarbonate solution and the mixture stirred for 2 h. Then the solid was filtered off, washed with water and dried at 40° C. to yield 18 mg of the title compound.

    [0521] LC/MS (Method LC5): RT=1.51 min; m/z=451.2 [M+H].sup.+

    Examples 42 and 43. 8-(4-[2-(4-Bromo-1H-pyrazol-1-yl)-ethoxy]-phenyl)-6-chloro-1-methyl-9H-pyrido[3,4-b]indole and 3-bromo-8-(4-[2-(4-bromo-1H-pyrazol-1-yl)-ethoxy]-phenyl)-6-chloro-1-methyl-9H-pyrido[3,4-b]indole

    [0522] Dry DMF (4 ml) was added to 8-(4-(2-(1H-pyrazol-1-yl)ethoxy)phenyl)-6-chloro-1-methyl-9H-pyrido[3,4-b]indole (52 mg), followed by N-bromosuccinimide (44 mg) The mixture was stirred for 3 h at 40° C. Further N-bromosuccinimide (22 mg) was added and the mixture stirred for another 3 h at 40° C. After standing overnight at room temperature, water was added and the aqueous phase was extracted three times with DCM. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by preparative RP HPLC. The fraction containing each of the two products were combined, the ACN was removed in vacuo and the residue was lyophilized.

    Example 42. 8-(4-[2-(4-Bromo-pyrazol-1-yl)-ethoxy]-phenyl)-6-chloro-1-methyl-9H-pyrido[3,4-b]indole

    [0523] ##STR00077##

    [0524] 7 mg of the title compound were obtained in the form of its salt with trifluoroacetic acid.

    [0525] LC/MS (Method LC5): RT=1.57 min; m/z=481.1 [M+H].sup.+

    Example 43. 3-Bromo-8-(4-[2-(4-bromo-pyrazol-1-yl)-ethoxy]-phenyl)-6-chloro-1-methyl-9H-pyrido[3,4-b]indole

    [0526] ##STR00078##

    [0527] 5 mg of the title compound were obtained in the form of its salt with trifluoroacetic acid.

    [0528] LC/MS (Method LC5): RT=2.13 min; m/z=559.0 [M+H].sup.+

    Example 44. 2-[4-(6-Chloro-9-ethyl-1-methyl-9H-pyrido[3,4-b]indol-8-yl)-pyrazol-1-yl]-ethanol

    [0529] ##STR00079##

    [0530] 8-(1-((1,3-Dioxolan-2-yl)methyl)-1H-pyrazol-4-yl)-6-chloro-9-ethyl-1-methyl-9H-pyrido[3,4-b]indole trifluoroacetic acid salt (55 mg) was stirred in a mixture of ACN (1.5 ml) and 2 N hydrochloric acid (0.5 ml) for 16 h. After removal of the solvent in vacuo the residue was purified by preparative RP HPLC. The fractions containing the product were combined, the ACN was removed in vacuo and the residue was lyophilized to yield 18 mg of the title compound in the form of its salt with trifluoroacetic acid.

    [0531] LC/MS (Method LC4): RT=1.16 min; m/z=355.2 [M+H].sup.+

    Example 45. 6-Chloro-1,5-dimethyl-8-[4-(2-pyrazol-1-yl-ethoxy)-phenyl]-9H-pyrido[3,4-b]pyridine

    [0532] ##STR00080##

    [0533] The title compound was synthesized analogously to the synthesis of the compound of example 8, using 185 mg of 6-chloro-8-iodo-1,5-dimethyl-9H-pyrido[3,4-b]indole and 170 mg of 1-(2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)ethyl)-1H-pyrazole and treating the reaction mixture in a microwave oven for 15 min at 100° C. After purification by HPLC, the residue of the extraction with EA was directly lyophilized, using water/ACN as the solvent. 129 mg of the title compound was obtained.

    [0534] LC/MS (Method LC5): RT=1.52 min; m/z=417.1 [M+H].sup.+

    Example 46. 6-Chloro-9-cyclopropylmethyl-8-(2,6-dichloro-pyridin-3-yl)-pyrido[3,4-b]indole

    [0535] ##STR00081##

    [0536] 8-Bromo-6-chloro-9-cyclopropylmethyl-pyrido[3,4-b]indole (3 g, 8.94 mmol) was dissolved in degassed DME (150 ml) and degassed water (48 ml). After addition of sodium carbonate (3.8 g, 35.75 mmol) the reaction mixture was flushed with argon. After heating to reflux, 2,6-dichloro-3-pyridinylboronic acid (3.4 g, 17.72 mmol) and BDFP (1.46 g, 1.79 mmol) were dissolved in dry DMF (45 ml), and the solution added to the reaction mixture via a syringe pump over 8 h. After 2.5 h an extra amount of 1.46 g (1.79 mmol) of BDFP was added to the reaction mixture. When the addition via the syringe pump was finished, the mixture was cooled, filtered, the precipitate washed with DCM and the filtrate concentrated in vacuo. The crude product was purified by preparative HPLC. The fractions containing the product were combined and lyophilized. 1.33 g of the title compound were obtained in the form of 6-chloro-9-cyclopropylmethyl-8-(2,6-dichloro-pyridin-3-yl)-pyrido[3,4-b]indole trifluoroacetic acid salt. This salt was dissolved in EA, and the solution washed with a saturated sodium hydrogencarbonate solution and water. The organic phase was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by chromatography over a 30 g SiO.sub.2 cartridge (EA:HEP 4:1). The fractions containing the product were concentrated in vacuo and the residue was treated with a HEP/EA mixture (15 ml, 4:1) and the mixture treated in a sonication bath. The solvent was removed in vacuo and the obtained solid dried under high vacuum to yield 711 mg of the title compound.

    [0537] LC/MS (Method LC3): RT=1.08 min; m/z=402.0 [M+H].sup.+

    Example 47. 5-(6-Chloro-9-cyclopropylmethyl-1-methyl-9H-pyrido[3,4-b]indol-8-yl)-pyridine-2-carbonitrile

    [0538] ##STR00082##

    [0539] Degassed dioxane (15 ml, abs.) and degassed water (4 ml) were charged in a 25 ml two-necked flask under argon. 8-Bromo-6-chloro-9-cyclopropylmethyl-1-methyl-pyrido[3,4-b]indole (300 mg, 0.86 mmol), sodium carbonate (272.8 mg, 2.57 mmol), 2-cyanopyridine-5-boronic acid pinacol ester (217.0 mg, 0.94 mmol) and BDFP (175.2 mg, 0.21 mmol) were added, and the mixture was heated under reflux for 12 h. EA (5 ml) was added, the reaction mixture was filtered through a kieselgur cartridge and eluted with EA (4×10 ml). The combined organic phases were concentrated and the residue was purified by preparative HPLC. 149 mg (36%) of the title compound was obtained in the form of its salt with trifluoroacetic acid.

    [0540] LC/MS (Method LC6): RT=1.10 min; m/z=373.2 [M+H].sup.+

    Example 48. 5-(6-Chloro-9-cyclopropylmethyl-1-methyl-9H-pyrido[3,4-b]indol-8-yl)-3-methyl-pyridine-2-carbonitrile

    [0541] ##STR00083##

    [0542] 8-Bromo-6-chloro-9-cyclopropylmethyl-1-methyl-pyrido[3,4-b]indole (300.0 mg, 0.86 mmol), cesium carbonate (559.0 mg, 1.72 mmol), BDFP (201.0 mg, 0.25 mmol) and 2-cyano-3-methylpyridine-5-boronic acid pinacol ester (419 mg, 1.72 mmol) were reacted and worked up analogously as described for the compound of example 47. 273 mg (64%) of the title compound was obtained in the form of its salt with trifluoroacetic acid.

    [0543] LC/MS (Method LC6): RT=1.13 min; m/z=387.1 [M+H].sup.+

    Example 49. 6-Chloro-9-cyclopropymethyl-1-methyl-8-(4-methyl-3,4-dihydro-2H-pyrido[3,2-b][1,4]oxazin-7-yl)-9H-pyrido[3,4-b]indole

    [0544] ##STR00084##

    [0545] 8-Bromo-6-chloro-9-cyclopropylmethyl-1-methyl-pyrido[3,4-b]indole (250.0 mg, 0.71 mmol), sodium carbonate (227.3 mg, 2.15 mmol), BDFP (146.0 mg, 0.18 mmol) and 4-methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydro-2H-pyrido[3,2-b][1,4]oxazine (197 mg, 0.72 mmol) were reacted and worked up analogously as described for the compound of example 47. 335 mg (88%) of the title compound was obtained in the form of its salt with trifluoroacetic acid.

    [0546] LC/MS (Method LC6): RT=1.09 min; m/z=419.2 [M+H].sup.+

    Example 50. 6-Chloro-9-cyclopropylmethyl-1-methyl-8-(6-pyrrolidin-1-ylpyridin-3-yl)-9H-pyrido[3,4-b]indole

    [0547] ##STR00085##

    [0548] 8-Bromo-6-chloro-9-cyclopropylmethyl-1-methyl-pyrido[34-b]indole (270 mg, 0.77 mmol), cesium carbonate (403 mg, 1.54 mmol), 2-(pyrrolidin-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (424 mg, 1.55 mmol) and BDFP (45 mg) were reacted and worked up analogously as described for the compound of example 47. 213 mg (52%) of the title compound was obtained in the form of its salt with trifluoroacetic acid.

    [0549] LC/MS (Method LC6): RT=1.01 min; m/z=417.2 [M+H].sup.+

    Example 51. 6-Chloro-9-cyclopropylmethyl-1-methyl-8-[6-(4-methyl-piperazin-1-yl)-pyridin-3-yl]-pyrido[3,4-b]indole

    [0550] ##STR00086##

    [0551] 8-Bromo-6-chloro-9-cyclopropylmethyl-1-methyl-pyrido[3,4-b]indole (270 mg, 0.77 mmol), cesium carbonate (504 mg, 1.55 mmol), 2-(4-methylpiperazin-1-yl)pyridine-5-boronic acid pinacol ester (469 mg, 1.55 mmol) and BDFP (181 mg, 0.22 mmol) were reacted and worked up analogously as described for the compound of example 47. 320 mg (74%) of the title compound was obtained in the form of its salt with trifluoroacetic acid.

    [0552] LC/MS (Method LC6): RT=0.93 min; m/z=446.2 [M+H].sup.+

    Example 52. 6-Chloro-8-(4-methoxy-phenyl)-9-(3-methyl-oxetan-3-ylmethyl)-9H-pyrido[3,4-b]indole

    [0553] ##STR00087##

    [0554] A microwave reaction vessel was charged with 8-bromo-6-chloro-9-(3-methyl-oxetan-3-ylmethyl)-9H-pyrido[3,4-b]indole (173 mg), sodium carbonate (201 mg), 2-(4-methoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (166 mg), BDFP (77 mg), DME (6 ml) and water (2 ml). After 12 min at 130° C. in a microwave oven the mixture was filtered and the filtrate concentrated in vacuo. The residue was first purified by chromatography over silica gel followed by a further purification by preparative RP HPLC. The fractions containing the product were combined, the ACN was removed in vacuo, the aqueous phase set to a basic pH with saturated sodium hydrogencarbonate solution, and extracted three times with EA. The combined organic phases were dried over sodium sulfate, filtered and concentrated in vacuo to yield 103 mg of the title compound.

    [0555] LC/MS (Method LC5): RT=1.70 min; m/z=393.3 [M+H].sup.+

    Example 53. 6-Chloro-5-(6-chloro-9-cyclopropylmethyl-9H-pyrido[3,4-b]indol-8-yl)-pyridin-2-ylamine

    [0556] ##STR00088##

    [0557] In a microwave vessel 6-chloro-9-cyclopropylmethyl-8-(2,6-dichloro-pyridin-3-yl)-pyrido[3,4-b]indole (20 mg) was dissolved in N-methyl-2-pyrrolidinone (0.5 ml), and an ammonia solution (0.1 ml, 25% in water) was added. After 2 h at 160° C. in a microwave oven further ammonia solution (0.1 ml) was added and heating was continued for 1.75 h at 200° C. After cooling, the mixture was concentrated in vacuo and the residue purified by preparative RP HPLC. The fractions containing the product were combined, the ACN was removed in vacuo and the residue was lyophilized to yield 30 mg of the title compound in the form of its salt with trifluoroacetic acid.

    [0558] LC/MS (Method LC5): RT=1.59 min; m/z=383.1 [M+H]

    [0559] The example compounds of the formula I listed in Table 1 were synthesized analogously to the syntheses of example compounds of the formula I described above. In Table 1, in the column “Ex. No.” the number of the example is given, in the column “LC/MS” the number of the HPLC method specified above which was used in the LC/MS characterization of the example compound is given, in the column “RT” the observed HPLC retention time in minutes is given, and in the column “MS” the mass-to-charge ratio m/z of the observed molecular ion or a related ion and the kind of the ion is given. Like in the case of the compounds of the formula I whose synthesis is described in detail above, the ionization method in the MS characterization was ES+ if the specified ion is [M+H].sup.+ or another positive ion, and ES− if the specified ion is [M−H].sup.− or another negative ion.

    TABLE-US-00001 TABLE 1 Ex. RT No. Structure Name LC/MS [min] MS  54 [00089]embedded image 6-Chloro-8-(6- chloro-pyridin-3- yl)-9-ethyl-9H- pyrido[3,4- b]indole LC10 2.77 342.0 [M + H].sup.+  55 [00090]embedded image 6-Chloro-8-(2,6- dichloro-pyridin-3- yl)-9-ethyl-9H- pyrido[3,4- b]indole LC9  2.10 376.1 [M + H].sup.+  56 [00091]embedded image 6-Chloro-8-(2- chloro-pyridin-3- yl)-9-ethyl-9H- pyrido[3,4- b]indole LC11 2.62 342.1 [M + H].sup.+  57 [00092]embedded image 6-Chloro-8- pyridin-3-yl-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC1  1.21 362.1 [M + H].sup.+  58 [00093]embedded image 6-Chloro-8-(2- chloro-pyridin-3- yl)-9-ethyl-1- methyl-9H- pyrido[3,4- b]indole LC1  1.36 356.1 [M + H].sup.+  59 [00094]embedded image 6-Chloro-8-(2,6- dichloro-pyridin-3- yl)-9-ethyl-1- methyl-9H- pyrido[3,4- b]indole LC6  1.15 390.1 [M + H].sup.+  60 [00095]embedded image 5-[6-Chloro-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4-b]indol- 8-yl]-pyridine-2- carbonitrile LC8  3.86 387.2 [M + H].sup.+  61 [00096]embedded image 6-Chloro-8- quinolin-3-yl-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC3  1.45 412.2 [M + H].sup.+  62 [00097]embedded image 6-Chloro-8-(6- methoxy-pyridin- 3-yl)-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4- b]indole LC11 2.85 392.1 [M + H].sup.+  63 [00098]embedded image 6-Chloro-8-(2,6- difluoro-pyridin-3- yl)-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4- b]indole LC11 2.84 398.2 [M + H].sup.+  64 [00099]embedded image 6-Chloro-8-(2,6- dichloro-pyridin-3- yl)-1-methyl-9H- pyrido[3,4- b]indole LC6  1.05 362.0 [M + H].sup.+  65 [00100]embedded image 6-Chloro-1- methyl-8-pyridin- 3-yl-9H- pyrido[3,4- b]indole LC11 2.12 294.2 [M + H].sup.+  66 [00101]embedded image 6-Chloro-8-(6- chloro-pyridin-3- yl)-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4- b]indole LC8  3.89 396.2 [M + H].sup.+  67 [00102]embedded image 6-Chloro-9-ethyl- 8-(6-methyl- pyridin-3-yl)-9H- pyrido[3,4- b]indole LC11 2.10 322.2 [M + H].sup.+  68 [00103]embedded image 6-Chloro-8-(6- methyl-pyridin-3- yl)-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4- b]indole LC11 2.22 376.2 [M + H].sup.+  69 [00104]embedded image 6-Chloro-8-(4- chloro-pyridin-3- yl)-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4- b]indole LC1  1.46 396.0 [M + H].sup.+  70 [00105]embedded image 6-Chloro-9-ethyl- 8-(6-methoxy- pyridin-3-yl)-9H- pyrido[3,4- b]indole LC11 2.77 338.2 [M + H].sup.+  71 [00106]embedded image 6-Chloro-8-(1H- pyrrolo[2,3- b]pyridin-5-yl)-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC4  1.52 401.2 [M + H].sup.+  72 [00107]embedded image 6-Chloro-8-(6- methoxy-pyridin- 3-yl)-9H- pyrido[3,4- b]indole LC6  1.35 310.1 [M + H].sup.+  73 [00108]embedded image 6-Chloro-8-(6- chloro-pyridin-3- yl)-9H-pyrido[3,4- b]indole LC1  1.34 314.0 [M + H].sup.+  74 [00109]embedded image 6-Chloro-8-(2,6- dichloro-pyridin-3- yl)-9H-pyrido[3,4- b]indole LC1  1.43 348.0 [M + H].sup.+  75 [00110]embedded image 6-Chloro-8-(6- methyl-pyridin-3- yl)-9H-pyrido[3,4- b]indole LC1  1.06 294.1 [M + H].sup.+  76 [00111]embedded image 6-Chloro-8-(5- chloro-thiophen-3- yl)-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4- b]indole LC6  1.29 401.1 [M + H].sup.+  77 [00112]embedded image 6-Chloro-8-(6- chloro-pyridin-3- yl)-1-methyl-9H- pyrido[3,4- b]indole LC3  1.03 328.0 [M + H].sup.+  78 [00113]embedded image 6-Chloro-8-(5- chloro-pyridin-3- yl)-9-ethyl-1- methyl-9H- pyrido[3,4- b]indole LC1  1.40 356.1 [M + H].sup.+  79 [00114]embedded image 6-Chloro-8-(6- chloro-pyridin-3- yl)-9-ethyl-1- methyl-9H- pyrido[3,4- b]indole LC3  1.09 356.0 [M + H].sup.+  80 [00115]embedded image (5-[6-Chloro-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4-b]indol- 8-yl]-pyridin-2-yl)- cyclohexyl-amine LC6  1.11 459.3 [M + H].sup.+  81 [00116]embedded image 6-Chloro-8-(6- pyrrolidin-1-yl- pyridin-3-yl)-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC12 2.68 431.1 [M + H].sup.+  82 [00117]embedded image 6-Chloro-8-(5- fluoro-pyridin-3- yl)-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4- b]indole LC12 3.29 380.0 [M + H].sup.+  83 [00118]embedded image 6-Chloro-8-(6- chloro-2-methyl- pyridin-3-yl)-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC12 3.62 410.0 [M + H].sup.+  84 [00119]embedded image (5-[6-Chloro-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4-b]indol- 8-yl]-pyridin-2-yl)- dimethyl-amine LC12 2.57 405.1 [M + H].sup.+  85 [00120]embedded image 6-Chloro-8-(5- fluoro-6-methoxy- pyridin-3-yl)-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC12 3.82 410.2 [M + H].sup.+  86 [00121]embedded image 6-Chloro-8-(6- morpholin-4-yl- pyridin-3-yl)-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC12 3.55 447.1 [M + H].sup.+  87 [00122]embedded image 6-Chloro-8-[6-(4- methyl-piperazin- 1-yl)-pyridin-3-yl]- 9-(2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC12 2.82 460.1 [M + H].sup.+  88 [00123]embedded image 6-Chloro-9-(2,2,2- trifluoro-ethyl)-8- (6-trifluoromethyl- pyridin-3-yl)-9H- pyrido[3,4- b]indole LC12 3.75 430.0 [M + H].sup.+  89 [00124]embedded image 6-Chloro-8- (3,4,5,6- tetrahydro-2H- [1,2′]bipyridinyl-5′- yl)-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4- b]indole LC4  1.72 445.2 [M + H].sup.+  90 [00125]embedded image 9-But-2-ynyl-6- chloro-8-(6- chloro-pyridin-3- yl)-9H-pyrido[3,4- b]indole LC12 3.35 366.1 [M + H].sup.+  91 [00126]embedded image 6-Chloro-9-ethyl- 8-(6-methoxy- pyridin-3-yl)-1- methyl-9H- pyrido[3,4- b]indole LC8  3.25 352.2 [M + H].sup.+  92 [00127]embedded image 6-Chloro-9-ethyl- 1-methyl-8-(1H- pyrrolo[2,3- b]pyridin-5-yl)-9H- pyrido[3,4- b]indole LC6  1.06 361.2 [M + H].sup.+  93 [00128]embedded image 6-Chloro-9-ethyl- 1-methyl-8-(6- morpholin-4-yl- pyridln-3-yl)-9H- pyrido[3,4- b]indole LC6  0.97 407.2 [M + H].sup.+  94 [00129]embedded image 6-Chloro-8-(6- methoxy-pyridin- 3-yl)-1-methyl-9H- pyrido[3,4- b]indole LC6  1.07 324.2 [M + H].sup.+  95 [00130]embedded image 6-Chloro-1- methyl-8-(6- methyl-pyridin-3- yl)-9H-pyrido[3,4- b]indole LC6  1.07 308.2 [M + H].sup.+  96 [00131]embedded image 6-Chloro-1- methyl-8-(6- morpholin-4-yl- pyridin-3-yl)-9H- pyrido[3,4- b]indole LC6  1.02 377.3 [M − H].sup.−  97 [00132]embedded image 6-Chloro-8-(4- chloro-pyridin-3- yl)-1-methyl-9H- pyrido[3,4- b]indole LC4  1.52 328.0 [M + H].sup.+  98 [00133]embedded image 6-Bromo-8-(2,6- dichloro-pyridin-3- yl)-1-methyl-9H- pyrido[3,4- b]indole LC8  3.21 406.1 [M + H].sup.+  99 [00134]embedded image 6-Chloro-8-(5- fluoro-6-methoxy- pyridin-3-yl)-1- methyl-9H- pyrido[3,4- b]indole LC6  1.09 342.1 [M + H].sup.+ 100 [00135]embedded image 5-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-pyridine-2- carbonitrile LC6  1.01 319.1 [M + H].sup.+ 101 [00136]embedded image 6-Chloro-1- methyl-8-(3,4,5,6- tetrahydro-2H- [1,2′]bipyridinyl-5′- yl)-9H-pyrido[3,4- b]indole LC6  1.32 377.3 [M + H].sup.+ 102 [00137]embedded image 6-Chloro-8-(6- isopropoxy- pyridin-3-yl)-1- methyl-9H- pyrido[3,4- b]indole LC3  1.02 352.1 [M + H].sup.+ 103 [00138]embedded image 6-Chloro-8-(2- chloro-6-methoxy- pyridin-3-yl)-1- methyl-9H- pyrido[3,4- b]indole LC3  0.99 358.1 [M + H].sup.+ 104 [00139]embedded image 6-Chloro-8-(6- methoxy-2- methyl-pyridin-3- yl)-1-methyl-9H- pyrido[3,4- b]indole LC3  0.98 338.1 [M + H].sup.+ 105 [00140]embedded image 6-Chloro-8-(2,6- dichloro-pyridin-3- yl)-1-ethyl-9H- pyrido[3,4- b]indole LC8  3.54 376.1 [M + H].sup.+ 106 [00141]embedded image 6-Chloro-8-(2,6- dichloro-pyridin-3- yl)-1-isopropyl- 9H-pyrido[3,4- b]indole LC3  1.04 390.1 [M + H].sup.+ 107 [00142]embedded image 5′-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)- [1,2′]bipyridinyl-2- one LC3  1.00 387.0 [M + H].sup.+ 108 [00143]embedded image 8-(2,6-Dichloro- pyridin-3-yl)-1,6- dimethyl-9H- pyrido[3,4- b]indole LC8  3.27 342.1 [M + H].sup.+ 109 [00144]embedded image 4-[5-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-pyridin-2-yl]- piperazin-2-one LC3  0.94 392.0 [M + H].sup.+ 110 [00145]embedded image 3-[5-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-pyridin-2-yl]- oxazolidin-2-one LC3  1.02 379.0 [M + H].sup.+ 111 [00146]embedded image 6-Chloro-8-(2,6- dichloro-pyridin-3- yl)-9-(2-methoxy- ethyl)-9H- pyrido[3,4- b]indole LC3  1.14 406.0 [M + H].sup.+ 112 [00147]embedded image 6-Chloro-8-(1- pyridin-4- ylmethyl-1H- pyrazol-4-yl)-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC3  1.01 442.0 [M + H].sup.+ 113 [00148]embedded image 3-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-6-methoxy- pyridin-2-ylamine LC8  3.03 339.1 [M + H].sup.+ 114 [00149]embedded image 6-Chloro-8-(2- chloro-3-methyl- 3H-imidazol-4-yl)- 9-(2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC3  1.12 399.1 [M + H].sup.+ 115 [00150]embedded image 6-Chloro-1- methyl-8-(1- pyridin-3- ylmethyl-1H- pyrazol-4-yl)-9H- pyrido[3,4- b]indole LC4  1.17 374.3 [M + H].sup.+ 116 [00151]embedded image 6-Chloro-9-ethyl- 1-methyl-8-(1- pyridin-3- ylmethyl-1H- pyrazol-4-yl)-9H- pyrido[3,4- b]indole LC4  1.22 402.3 [M + H].sup.+ 117 [00152]embedded image 6-Chloro-8-(2,6- dichloro-pyridin-3- yl)-1-methyl-9H- pyrido[3,4- b]indole-3- carboxylic acid methyl ester LC4  2.06 420.1 [M + H].sup.+ 118 [00153]embedded image 6-Chloro-1- methyl-8-[6-(4- methyl-piperazin- 1-yl)-pyridin-3-yl]- 9H-pyrido[3,4- b]indole LC4  1.24 390.4 [M − H].sup.− 119 [00154]embedded image 8-(2-Benzyloxy- pyridin-3-yl)-6- chloro-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.73 400.1 [M + H].sup.+ 120 [00155]embedded image 6-Chloro-1- methyl-8-(6- methylsulfanyl- pyridin-3-yl)-9H- pyrido[3,4- b]indole LC4  1.45 340.1 [M + H].sup.+ 121 [00156]embedded image 6-Chloro-8-[1- (2,6-dimethyl- pyridin-3- ylmethyl)-1H- pyrazol-4-yl)-9- ethyl-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.08 430.3 [M + H].sup.+ 122 [00157]embedded image 6-Chloro-8-(6- chloro-2-methoxy- pyridin-3-yl)-1- methyl-9H- pyrido(3,4- b]indole LC4  1.52 358.2 [M + H].sup.+ 123 [00158]embedded image 6-Chloro-9-ethyl- 8-pyridin-4-yl-9H- pyrido[3,4- b]indole LC9  1.49 308.2 [M + H].sup.+ 124 [00159]embedded image 6-Chloro-9-ethyl- 8-furan-2-yl-9H- pyrido[3,4- b]indole LC2  1.35 297.1 [M + H].sup.+ 125 [00160]embedded image 6-Chloro-9-ethyl- 1-methyl-8- pyridin-4-yl-9H- pyrido[3,4- b]indole LC3  0.95 322.1 [M + H].sup.+ 126 [00161]embedded image 6-Chloro-8- pyrimidin-5-yl-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC1  1.30 363.1 [M + H].sup.+ 127 [00162]embedded image 6-Chloro-8- pyridin-4-yl-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC1  1.20 362.1 [M + H].sup.+ 128 [00163]embedded image 6-Chloro-8- phenyl-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4- b]indole LC8  4.54 361.0 [M + H].sup.+ 129 [00164]embedded image 6-Chloro-8-(4- chloro-phenyl)-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC3  1.35 394.9 [M + H].sup.+ 130 [00165]embedded image 6-Chloro-1- methyl-8-pyridin- 4-yl-9H- pyrido[3,4- b]indole LC3  0.86 294.0 [M + H].sup.+ 131 [00166]embedded image 6-Chloro-8-(4- chloro-phenyl)-9- ethyl-9H- pyrido[3,4- b]indole LC11 3.10 341.2 [M + H].sup.+ 132 [00167]embedded image 5-[6-Chloro-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4-b]indol- 8-yl]-pyrimidin-2- ylamine LC11 2.39 378.2 [M + H].sup.+ 133 [00168]embedded image 6-Chloro-9-ethyl- 8-pyrimidin-5-yl- 9H-pyrido[3,4- b]indole LC1  1.20 309.1 [M + H].sup.+ 134 [00169]embedded image 6-Chloro-8-(5- chloro-thiophen-2- yl)-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4- b]indole LC12 4.22 401.0 [M + H].sup.+ 135 [00170]embedded image 6-Chloro-8-(1- methyl-1H- pyrazol-4-yl)-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC12 3.17 365.1 [M + H].sup.+ 136 [00171]embedded image 6-Chloro-8-(3- phenyl-isoxazol-5- yl)-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4- b]indole LC8  4.60 428.2 [M + H].sup.+ 137 [00172]embedded image 6-Chloro-8-(1- isobutyl-1H- pyrazol-4-yl)-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC12 3.79 407.1 [M + H].sup.+ 138 [00173]embedded image 6-Chloro-9-(2,2,2- trifluoro-ethyl)-8- (1,3,5-trimethyl- 1H-pyrazol-4-yl)- 9H-pyrido[3,4- b]indole LC12 3.40 393.1 [M + H].sup.+ 139 [00174]embedded image 6-Chloro-8- pyrazolo[1,5- a]pyridin-3-yl-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC12 3.47 401.1 [M + H].sup.+ 140 [00175]embedded image 6-Chloro-8-(2,5- dimethyl-2H- pyrazol-3-yl)-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC3  1.16 379.0 [M + H].sup.+ 141 [00176]embedded image 6-Chloro-8- pyrimidin-5-yl-9H- pyrido[3,4- b]indole LC6  0.88 281.1 [M + H].sup.+ 142 [00177]embedded image 6-Chloro-9-ethyl- 8-(4-methoxy- phenyl)-9H- pyrido[3,4- b]indole LC6  1.14 337.20 [M + H].sup.+ 143 [00178]embedded image 6-Chloro-9-ethyl- 8-p-tolyl-9H- pyrido[3,4- b]indole LC6  1.22 321.1 [M + H].sup.+ 144 [00179]embedded image 6-Chloro-8-(1H- pyrazol-4-yl)-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC12 3.09 351.1 [M + H].sup.+ 145 [00180]embedded image 6-Chloro-8-(2- chloro-phenyl)-9- ethyl-9H- pyrido[3,4- b]indole LC8  3.96 341.1 [M + H].sup.+ 146 [00181]embedded image 6-Chloro-9-ethyl- 8-(3-methoxy- phenyl)-9H- pyrido[3,4- b]indole LC8  3.91 337.1 [M + H].sup.+ 147 [00182]embedded image 6-Chloro-9-ethyl- 8-(4-methoxy- phenyl)-1-methyl- 9H-pyrido[3,4- b]indole LC8  3.70 351.2 [M + H].sup.+ 148 [00183]embedded image 6-Chloro-8-(4- chloro-phenyl)-9- ethyl-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.64 355.1 [M + H].sup.+ 149 [00184]embedded image 6-Chloro-8-(4- methoxy-phenyl)- 1-methyl-9H- pyrido[3,4- b]indole LC3  1.10 323.1 [M + H].sup.+ 150 [00185]embedded image 6-Chloro-1- methyl-8-p-tolyl- 9H-pyrido[3,4- b]indole LC6  1.15 305.3 [M − H].sup.− 151 [00186]embedded image 6-Chloro-1- methyl-8-(1- methyl-1H- pyrazol-4-yl)-9H- pyrido[3,4- b]indole LC8  2.80 297.2 [M + H].sup.+ 152 [00187]embedded image 6-Chloro-8-(4- methoxy-phenyl)- 1-methyl-9H- pyrido[3,4- b]indole-3- carboxylic acid methyl ester LC6  1.36 381.1 [M + H].sup.+ 153 [00188]embedded image 6-Chloro-1-ethyl- 8-(4-methoxy- phenyl)-9H- pyrido[3,4- b]indole LC3  1.01 337.1 [M + H].sup.+ 154 [00189]embedded image 5-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-pyrimidin-2- ylamine LC3  0.80 310.1 [M + H].sup.+ 155 [00190]embedded image 6-Chloro-8-(4- methoxy-phenyl)- 1-trifluoromethyl- 9H-pyrido[3,4- b]indole LC3  1.32 377.1 [M + H].sup.+ 156 [00191]embedded image 6-Chloro-1- methyl-8-(2- pyrrol-1-yl- pyrimidin-5-yl)- 9H-pyrido[3,4- b]indole LC3  1.01 360.1 [M + H].sup.+ 157 [00192]embedded image [5-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-pyrimidin-2- yl]-methyl-amine LC6  0.99 324.1 [M + H].sup.+ 158 [00193]embedded image 2,6-Dichloro-3-(6- chloro-1-methyl- 9H-pyrido[3,4- b]indol-8-yl)- benzonitrile LC8  3.64 386.1 [M + H].sup.+ 159 [00194]embedded image 6-Chloro-8-(1- ethyl-1H-pyrazol- 4-yl)-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4- b]indole LC3  1.12 379.0 [M + H].sup.+ 160 [00195]embedded image 6-Chloro-8-(1- isopropyl-1H- pyrazol-4-yl)-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC3  1.18 393.0 [M + H].sup.+ 161 [00196]embedded image 6-Chloro-8-(1- propyl-1H- pyrazol-4-yl)-9- (2,2,2-trifIuoro- ethyl)-9H- pyrido[3,4- b]indole LC3  1.16 393.0 [M + H].sup.+ 162 [00197]embedded image 8-(1-Benzyl-1H- pyrazol-4-yl)-6- chloro-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4- b]indole LC8  3.91 441.2 [M + H].sup.+ 163 [00198]embedded image Acetic acid 2-(4- [6-chloro-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4-b]indol- 8-yl]-pyrazol-1-yl)- ethyl ester LC3  1.12 437.0 [M + H].sup.+ 164 [00199]embedded image 6-Chloro-9-ethyl- 1-methyl-8-(1- methyl-1H- pyrazol-4-yl)-9H- pyrido[3,4- b]indole LC8  2.87 325.1 [M + H].sup.+ 165 [00200]embedded image 6-Chloro-8-(1- methyl-1H-pyrrol- 3-yl)-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4- b]indole LC3  1.25 364.0 [M + H].sup.+ 166 [00201]embedded image 6-Chloro-8-(1- thiophen-2- ylmethyl-1H- pyrazol-4-yl)-9- (2,2,2-trifiuoro- ethyl)-9H- pyrido[3,4- b]indole LC3  1.21 491.1 [M − H + FA].sup.− 167 [00202]embedded image 6-Chloro-8-[1- (2,6-dichloro- phenyl)-1H- pyrazol-4-yl]-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC3  1.28 495.1 [M + H].sup.+ 168 [00203]embedded image 6-Chloro-8-(1- pyridin-2- ylmethyl-1H- pyrazol-4-yl)-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC3  1.10 442.1 [M + H].sup.+ 169 [00204]embedded image 6-Chloro-8-[1-(2- methoxy-ethyl)- 1H-pyrazol-4-yl]- 9-(2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC3  1.09 409.2 [M + H].sup.+ 170 [00205]embedded image 6-Chloro-8-(2- phenyl-thiazol-5- yl)-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4- b]indole LC3  1.37 444.1 [M + H].sup.+ 171 [00206]embedded image 6-Chloro-8-(2- methyl-thiazol-5- yl)-9-(2,2,2- trifluoro-ethyl)-9H- pyrido[3,4- b]indole LC3  1.17 382.1 [M + H].sup.+ 172 [00207]embedded image 6-Chloro-8-(3,4- dimethoxy- phenyl)-1-methyl- 9H-pyrido[3,4- b]indole LC3  0.95 353.3 [M + H].sup.+ 173 [00208]embedded image 6-Chloro-8-(1- cyclopropylmethyl- 1H-pyrazol-4-yl)- 9-(2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC3  1.18 405.2 [M + H].sup.+ 174 [00209]embedded image 6-Chloro-8-(2- cyclopropyl- thiazol-5-yl)-9- (2,2,2-trifluoro- ethyl)-9H- pyrido[3,4- b]indole LC3  1.26 408.2 [M + H].sup.+ 175 [00210]embedded image 6-Chloro-1- methyl-8-(2,4,6- trimethoxy- phenyl)-9H- pyrido[3,4- b]indole LC3  1.10 383.2 [M + H].sup.+ 176 [00211]embedded image 6-Chloro-8-(4- isopropoxy-2- methyl-phenyl)-1- methyl-9H- pyrido[3,4- b]indole LC8  4.05 365.2 [M + H].sup.+ 177 [00212]embedded image 8-(4-Benzyloxy- phenyl)-6-chloro- 1-methyl-9H- pyrido[3,4- b]indole LC8  4.13 399.1 [M + H].sup.+ 178 [00213]embedded image 6-Chloro-1- methyl-8-(2,3,4- trimethoxy- phenyl)-9H- pyrido[3,4- b]indole LC4  1.64 381.2 [M − H].sup.− 179 [00214]embedded image 6-Chloro-8-(2,3- dihydro- benzofuran-5-yl)- 1-methyl-9H- pyrido(3,4- b]indole LC4  1.64 335.2 [M + H].sup.+ 180 [00215]embedded image 5-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-2-methoxy- phenol LC4  1.54 339.2 [M + H].sup.+ 181 [00216]embedded image 6-Chloro-8-(4- cyclopropyl- methoxy-phenyl)- 1- methyl-9H- pyrido[3,4- b]indole LC4  1.77 363.2 [M + H].sup.+ 182 [00217]embedded image 6-Chloro-1- methyl-8-[4-(3- methyl-oxetan-3- ylmethoxy)- phenyl]-9H- pyrido[3,4- b]indole LC4  1.67 393.2 [M + H].sup.+ 183 [00218]embedded image 6-Chloro-8-(4- cyclopropoxy- phenyl)-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.73 349.2 [M + H].sup.+ 184 [00219]embedded image 6-Chloro-8- chroman-6-yl-1- methyl-9H- pyrido[3,4- b]indole LC4  1.69 349.2 [M + H].sup.+ 185 [00220]embedded image 8-(3-Benzyloxy-4- methoxy-phenyl)- 6-chloro-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.65 429.3 [M + H].sup.+ 186 [00221]embedded image 5-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-2-methoxy- phenylamine LC4  1.52 338.2 [M + H].sup.+ 187 [00222]embedded image 6-Chloro-8-(4- methoxy-2,3- dimethyl-phenyl)- 1-methyl-9H- pyrido[3,4- b]indole LC4  1.75 351.2 [M + H].sup.+ 188 [00223]embedded image 6-Bromo-8-(4- methoxy-phenyl)- 1-methyl-9H- pyrido[3,4- b]indole LC3  1.10 367.1 [M + H].sup.+ 189 [00224]embedded image 6-Chloro-8-(4- chloro-phenyl)-1- methyl-9H- pyrido[3,4- b]indole LC4  1.70 327.2 [M + H].sup.+ 190 [00225]embedded image 6-Chloro-8-(2,4- dichloro-phenyl)- 1-methyl-9H- pyrido[3,4- b]indole LC4  1.61 361.2 [M + H].sup.+ 191 [00226]embedded image 6-Chloro-8-(4- methoxy-phenyl)- 9H-pyrido[3,4- b]indole LC4  1.48 309.3 [M + H].sup.+ 192 [00227]embedded image 6-Chloro-8-(4- ethoxy-phenyl)-1- methyl-9H- pyrido[3,4- b]indole LC4  1.57 337.31 [M + H].sup.+ 193 [00228]embedded image 6-Chloro-8-(2,4- dimethoxy- phenyl)-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.67 353.3 [M + H].sup.+ 194 [00229]embedded image 6-Chloro-8-[4-(2- imidazol-1-yl- ethoxy)-phenyl]-1- methyl-9H- pyrido[3,4- b]indole LC4  1.13 403.3 [M + H].sup.+ 195 [00230]embedded image 6-Chloro-8-(2,2- dimethyl-2,3- dihydro- benzofuran-6-yl)- 1-methyl-9H- pyrido[3,4- b]indole LC4  1.76 363.2 [M + H].sup.+ 196 [00231]embedded image 6-Chloro-1- methyl-8-(2- methyl-2,3- dihydro- benzofuran-6-yl)- 9H-pyrido[3,4- b]indole LC4  1.56 349.2 [M + H].sup.+ 197 [00232]embedded image 6-Chloro-1- methyl-8-[4- (tetrahydro-furan- 3-yloxy)-phenyl]- 9H-pyrido[3,4- b]indole LC4  1.64 379.3 [M + H].sup.+ 198 [00233]embedded image 6-Chloro-1- methyl-8-[3- methyl-4- (tetrahydrofuran- 3-yloxy)-phenyl]- 9H-pyrido[3,4- b]indole LC4  1.72 393.3 [M + H].sup.+ 199 [00234]embedded image 8-(3-Benzyl-1H- pyrazol-4-yl)-6- chloro-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.57 373.2 [M + H].sup.+ 200 [00235]embedded image 6-Chloro-8-(4- methoxy-3- methoxymethyl- phenyl)-1-methyl- 9H-pyrido[3,4- b]indole LC8  3.53 367.2 [M + H].sup.+ 201 [00236]embedded image 6-Chloro-9-(2- methoxy-ethyl)-8- (4-methoxy- phenyl)-9H- pyrido[3,4- b]indole LC4  1.71 367.2 [M + H].sup.+ 202 [00237]embedded image 6-Chloro-1- methyl-8-[4- (pyridin-2- ylmethoxy)- phenyl]-9H- pyrido[3,4- b]indole LC8  3.30 400.2 [M + H].sup.+ 203 [00238]embedded image 2-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-1H-indole-5- carboxylic acid ethyl ester LC4  1.71 402.3 [M − H].sup.− 204 [00239]embedded image 2-[4-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-phenyl]-1-(4- pyrrolidin-1-yl- piperidin-1-yl)- ethanone LC8  2.51 487.3 [M + H].sup.+ 205 [00240]embedded image 6-Chloro-1- methyl-8-(4- phenoxy-phenyl)- 9H-pyrido[3,4- b]indole (a) LC4  1.68 385.2 [M + H].sup.+ 206 [00241]embedded image 6-Chloro-8-(3- chloro-4-methoxy- phenyl)-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.70 357.2 [M + H].sup.+ 207 [00242]embedded image 8-(1-Benzyl-1H- pyrazol-4-yl)-6- chloro-9-ethyl-1- methyl-9H- pyrido[3,4- b]indole LC4  1.57 401.2 [M + H].sup.+ 208 [00243]embedded image 8-(1-Benzyl-1H- pyrazol-4-yl)-6- bromo-9-ethyl-1- methyl-9H- pyrido[3,4- b]indole LC4  1.58 445.2 [M + H].sup.+ 209 [00244]embedded image 6-Bromo-9-ethyl- 1-methyl-8-(1- methyl-1H- pyrazol-4-yl)-9H- pyrido[3,4- b]indole LC4  1.34 369.1 [M + H].sup.+ 210 [00245]embedded image 6-Bromo-1,9- diethyl-3-methyl- 8-(1-methyl-1H- pyrazol-4-yl)-9H- pyrido[3,4- b]indole LC4  1.55 397.1 [M + H].sup.+ 211 [00246]embedded image 6-Bromo-3-ethyl- 8-(4-methoxy- phenyl)-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.80 395.1 [M + H].sup.+ 212 [00247]embedded image 6-Bromo-3-ethyl- 1-methyl-8-[4-(2- pyrazol-1-yl- ethoxy)-phenyl]- 9H-pyrido[3,4- b]indole LC4  1.60 475.2 [M + H].sup.+ 213 [00248]embedded image 6-Chloro-1- methyl-8-(2- piperazin-1-yl- pyridin-4-yl)-9H- pyrido[3,4- b]indole LC4  1.03 376.1 [M − H].sup.− 214 [00249]embedded image 6-Chloro-1- methyl-8-[2-(4- methyl-piperazin- 1-yl)-pyridin-4-yl]- 9H-pyrido[3,4- b]indole LC4  1.04 390.1 [M − H].sup.− 215 [00250]embedded image 4-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-pyridin-2- ylamine LC4  1.06 309.2 [M + H].sup.+ 216 [00251]embedded image 6-Bromo-3,9- diethyl-1-methyl- 8-(1-methyl-1H- pyrazol-4-yl)-9H- pyrido[3,4- b]indole LC4  1.43 397.2 [M + H].sup.+ 217 [00252]embedded image 6-Bromo-8-(4- methoxy-phenyl)- 1,3-dimethyl-9H- pyrido[3,4- b]indole LC4  1.56 381.1 [M + H].sup.+ 218 [00253]embedded image 6-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-3-methoxy- pyridin-2-ylamine LC4  1.51 339.2 [M + H].sup.+ 219 [00254]embedded image 6-Chloro-9-ethyl- 1-methyl-8-[2-(4- methyl-piperazin- 1-yl)-pyridin-4-yl]- 9H-pyrido[3,4- b]indole LC4  1.24 420.2 [M + H].sup.+ 220 [00255]embedded image 3-(6-Chloro-9- ethyl-1-methyl- 9H-pyrido[3,4- b]indol-8-yl)- benzamide LC8  2.88 364.1 [M + H].sup.+ 221 [00256]embedded image 6-Chloro-8-[3- methoxy-4- (pyridin-2- ylmethoxy)- phenyl]-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.62 428.2 [M − H].sup.− 222 [00257]embedded image 8-(4-Benzyloxy-3- methoxy-phenyl)- 6-chloro-1-methyl- 9H-pyrido[3,4- b]indole LC8  3.98 429.2 [M + H].sup.+ 223 [00258]embedded image 6-Chloro-8-[4-(6- fluoro-pyridin-2- ylmethoxy)-3- methoxy-phenyl]- 1-methyl-9H- pyrido[3,4- b]indole LC4  1.71 446.2 [M − H].sup.− 224 [00259]embedded image 6-Chloro-1- methyl-8-(4- morpholin-4- ylmethyl-phenyl)- 9H-pyrido[3,4- b]indole LC8  2.29 392.1 [M + H].sup.+ 225 [00260]embedded image 6-Chloro-8-(4- cyclopentyloxy- phenyl)-1-methyl- 9H-pyrido(3,4- b]indole LC8  4.18 377.2 [M + H].sup.+ 226 [00261]embedded image 6-Chloro-8-[4-(6- fluoro-pyridin-2- ylmethoxy)- phenyl]-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.75 418.1 [M + H].sup.+ 227 [00262]embedded image 6-Chloro-1- methyl-8-(2- methyl-2H- pyrazol-3-yl)-9H- pyrido[3,4- b]indole LC4  1.44 295.2 [M − H].sup.− 228 [00263]embedded image 6-Chloro-1- methyl-8-[4-(1- methyl-pyrrolidin- 3-ylmethoxy)- phenyl]-9H- pyrido[3,4- b]indole LC4  1.36 406.2 [M + H].sup.+ 229 [00264]embedded image 1-[4-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-phenoxy]-3- piperidin-1-yl- propan-2-ol LC4  1.36 450.1 [M + H].sup.+ 230 [00265]embedded image 1-[3-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-phenyl]-3- (2,4-difluoro- phenyl)-urea LC4  1.72 463.1 [M + H].sup.+ 231 [00266]embedded image 6-Chloro-1- methyl-8-(4- phenethyloxy- phenyl)-9H- pyrido[3,4- b]indole LC4  1.87 413.1 [M + H].sup.+ 232 [00267]embedded image 6-Chloro-8-(1H- indazol-5-yl)-1- methyl-9H- pyrido[3,4- b]indole LC4  1.53 333.1 [M + H].sup.+ 233 [00268]embedded image 6-Chloro-9-ethyl- 1-methyl-8-[1-(2- pyrazol-1-yl- ethyl)-1H-pyrazol- 3-yl]-9H- pyrido(3,4- b]indole LC4  1.35 405.2 [M + H].sup.+ 234 [00269]embedded image 6-Chloro-9-ethyl- 1-methyl-8-(1H- pyrazol-3-yl)-9H- pyrido[3,4- b]indole LC8  2.83 311.0 [M + H].sup.+ 235 [00270]embedded image 6-Chloro-9-ethyl- 1-methyl-8-[1-(2- pyrazol-1-yl- ethyl)-1H-pyrazol- 3-yl]-9H- pyrido[3,4- b]indole LC4  1.49 405.1 [M + H].sup.+ 236 [00271]embedded image 6-Chloro-8-(2,2- dimethyl-2,3- dihydro- benzofuran-5-yl)- 1-methyl-9H- pyrido[3,4- b]indole LC4  1.61 363.2 [M + H].sup.+ 237 [00272]embedded image 6-Chloro-1- methyl-8-(2- pyrazol-1-yl- pyrimidin-5-yl)- 9H-pyrido[3,4- b]indole LC4  1.31 361.14 [M + H].sup.+ 238 [00273]embedded image [4-(6-Chloro-9- ethyl-1-methyl- 9H-pyrido[3,4- b]indol-8-yl)- phenyl]-phenyl- methanol LC4  1.58 427.2 [M + H].sup.+ 239 [00274]embedded image 6-Chloro-9-ethyl- 1-methyl-8-(1H- pyrazol-4-yl)-9H- pyrido[3,4- b]indole LC4  1.20 311.1 [M + H].sup.+ 240 [00275]embedded image 6-(6-Chloro-1,9- dimethyl-9H- pyrido[3,4-b]indol- 8-yl)-3-methoxy- pyridin-2-ylamine LC4  1.17 353.1 [M + H].sup.+ 241 [00276]embedded image 4-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-benzylamine LC7  4.51 161.6 [M + 2H].sup.++ 242 [00277]embedded image 6-Chloro-1- methyl-8-(1- pyridin-2- ylmethyl-1H- pyrazol-4-yl)-9H- pyrido[3,4- b]indole LC4  1.32 374.2 [M + H].sup.+ 243 [00278]embedded image 6-Chloro-8-(4- methanesulfinyl- phenyl)-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.27 355.1 [M + H].sup.+ 244 [00279]embedded image 6-Chloro-9-ethyl- 1-methyl-8-(1- pyridin-2- ylmethyl-1H- pyrazol-4-yl)-9H- pyrido[3,4- b]indole LC4  1.34 402.2 [M + H].sup.+ 245 [00280]embedded image 8-(3-Bromo-2- fluoro-pyridin-4- yl)-6-chloro-1- methyl-9H- pyrido[3,4- b]indole LC4  1.44 390.0 [M + H].sup.+ 246 [00281]embedded image 6-Chloro-9-ethyl- 1-methyl-8-(4- phenoxy-phenyl)- 9H-pyrido[3,4- b]indole LC4  1.74 413.2 [M + H].sup.+ 247 [00282]embedded image 6-Chloro-1- methyl-8-(3- morpholin-4-yl- phenyl)-9H- pyrido[3,4- b]indole LC4  1.48 378.2 [M + H].sup.+ 248 [00283]embedded image 2-[3-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-phenyl]- ethanol LC4  1.39 337.2 [M + H].sup.+ 249 [00284]embedded image 6-Chloro-1- methyl-8-[3-(2- morpholin-4-yl- ethoxy)-phenyl]- 9H-pyrido[3,4- b]indole LC4  1.11 422.1 [M + H].sup.+ 250 [00285]embedded image 3-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)- benzaldehyde LC4  1.40 321.2 [M + H].sup.+ 251 [00286]embedded image 8-(3,5-Bis- trifluoromethyl- phenyl)-6-chloro- 1-methyl-9H- pyrido[3,4- b]indole LC4  1.69 429.1 [M + H].sup.+ 252 [00287]embedded image 6-Chloro-1- methyl-8-(4- trifluoromethyl- phenyl)-9H- pyrido[3,4- b]indole LC4  1.60 361.1 [M + H].sup.+ 253 [00288]embedded image 6-Chloro-9-ethyl- 1-methyl-8-(3- morpholin-4-yl- phenyl)-9H- pyrido[3,4- b]indole LC4  1.54 406.3 [M + H].sup.+ 254 [00289]embedded image 6-Chloro-9-ethyl- 1-methyl-8-(3- morpholin-4- ylmethyl-phenyl)- 9H-pyrido[3,4- b]indole LC4  1.06 420.3 [M + H].sup.+ 255 [00290]embedded image 6-Chloro-1- methyl-8-(3- morpholin-4- ylmethyl-phenyl)- 9H-pyrido[3,4- b]indole LC4  1.01 392.2 [M + H].sup.+ 256 [00291]embedded image 6-Chloro-9-ethyl- 1-methyl-8-[4-(3- methyl-oxetan-3- ylmethoxy)- phenyl]-9H- pyrido[3,4- b]indole LC4  1.57 421.3 [M + H].sup.+ 257 [00292]embedded image 6-Bromo-9-ethyl- 1-methyl-8-[4-(3- methyl-oxetan-3- ylmethoxy)- phenyl]-9H- pyrido[3,4- b]indole LC4  1.63 465.1 [M + H].sup.+ 258 [00293]embedded image [3-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-phenyl]- pyridin-2-yl- methanol LC4  1.29 398.1 [M − H].sup.− 259 [00294]embedded image 6-Bromo-1- methyl-8-[4-(2- pyrazol-1-yl- ethoxy)-phenyl]- 9H-pyrido[3,4- b]indole LC4  1.49 447.3 [M + H].sup.+ 260 [00295]embedded image 6-Chloro-1- methyl-8-[3- (quinolin-2- ylmethoxy)- phenyl]-9H- pyrido[3,4- b]indole LC4  1.61 450.3 [M + H].sup.+ 261 [00296]embedded image 6-Bromo-9-ethyl- 1-methyl-8-(1- pyridin-2- ylmethyl-1H- pyrazol-4-yl)-9H- pyrido[3,4- b]indole LC4  1.34 446.3 [M + H].sup.+ 262 [00297]embedded image 6-Chloro-1- methyl-8-(4- trifluoromethoxy- phenyl)-9H- pyrido[3,4- b]indole LC4  1.61 377.2 [M + H].sup.+ 263 [00298]embedded image Acetic acid 2-[4- (6-chloro-9-ethyl- 1-methyl-9H- pyrido[3,4-b]indol- 8-yl)-pyrazol-1-yl]- ethyl ester LC14 0.83 397.2 [M + H].sup.+ 264 [00299]embedded image 4-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-N-(2- dimethylamino- ethyl)-benzamide LC4  1.03 407.2 [M + H].sup.+ 265 [00300]embedded image 3-(6-Chloro-1- methyl-9H- pyrido[3,4-b]indol- 8-yl)-N-(2- dimethylamino- ethyl)-benzamide LC4  1.01 407.2 [M + H].sup.+ 266 [00301]embedded image 6-Chloro-1- methyl-8-[4- (pyrimidin-2- ylmethoxy)- phenyl]-9H- pyrido[3,4- b]indole LC4  1.38 401.2 [M + H].sup.+ 267 [00302]embedded image 6-Chloro-1- methyl-8-[3- (pyrimidin-2- ylmethoxy)- phenyl]-9H- pyrido[3,4- b]indole LC4  1.42 401.2 [M + H].sup.+ 268 [00303]embedded image 8-[4-(5-Bromo- pyrimidin-2- yloxy)-phenyl]-6- chloro-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.53 465.2 [M + H].sup.+ 269 [00304]embedded image 6-Chloro-8-[3- (isoquinolin-1- ylmethoxy)- phenyl]-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.57 450.2 [M + H].sup.+ 270 [00305]embedded image 6-Chloro-8-[4- (isoquinolin-1- ylmethoxy)- phenyl]-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.57 450.2 [M + H].sup.+ 271 [00306]embedded image 6-Chloro-1- methyl-8-[3-(1- methyl-1H- imidazol-2- ylmethoxy)- phenyl]-9H- pyrido[3,4- b]indole LC4  1.14 401.2 [M − H].sup.− 272 [00307]embedded image 6-Chloro-8-[3- (4,6-dimethoxy- pyrimidin-2- ylmethoxy)- phenyl]-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.58 461.2 [M + H].sup.+ 273 [00308]embedded image 6-Chloro-1- methyl-8-[3- (thiazol-2- ylmethoxy)- phenyl]-9H- pyrido[3,4- b]indole LC4  1.51 406.2 [M + H].sup.+ 274 [00309]embedded image 6-Chloro-1- methyl-8-[3- (quinazolin-2- ylmethoxy)- phenyl]-9H- pyrido[3,4- b]indole LC4  1.51 451.4 [M + H].sup.+ 275 [00310]embedded image 6-Chloro-8-[4- (4,6-dimethoxy- pyrimidin-2- ylmethoxy)- phenyl]-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.58 461.3 [M + H].sup.+ 276 [00311]embedded image 6-Chloro-8-(1- [1,3]dioxolan-2- ylmethyl-1H- pyrazol-4-yl)-9- ethyl-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.30 397.2 [M + H].sup.+ 277 [00312]embedded image 6-Chloro-1- methyl-8-[3-(5- trifluoromethyl- furan-2- ylmethoxy)- phenyl]-9H- pyrido[3,4- b]indole LC8  4.01 457.3 [M + H].sup.+ 278 [00313]embedded image 6-Chloro-8-[4- (2,3-dihydro- benzo[1,4]dioxin- 2-ylmethoxy)- phenyl]-1-methyl- 9H-pyrido[3,4- b]indole LC4  1.66 457.3 [M + H].sup.+ 279 [00314]embedded image 6-Chloro-8-(6- chloro-pyridin-3- yl)-9- cyclobutylmethyl- 9H-pyrido[3,4- b]indole LC6  1.08 382.1 [M + H].sup.+ 280 [00315]embedded image 6-Chloro-9- cyclopropylmethyl- 8-pyridin-3-yl-9H- pyrido[3,4- b]indole LC6  0.92 334.1 [M + H].sup.+ 281 [00316]embedded image 6-Chloro-9- cyclopropylmethyl- 1-methyl-8- pyrimidin-5-yl-9H- pyrido[3,4- b]indole LC8  2.82 349.2 [M + H].sup.+ 282 [00317]embedded image 6-Chloro-9- cyclopropylmethyl- 8-(2,4-dichloro- phenyl)-1-methyl- 9H-pyrido[3,4- b]indole LC6  1.24 415.1 [M + H].sup.+ 283 [00318]embedded image 6-Chloro-8-(4- chloro-pyridin-3- yl)-9- cyclopropylmethyl- 1-methyl-9H- pyrido[3,4- b]indole LC6  1.08 382.1 [M + H].sup.+ 284 [00319]embedded image 6-Chloro-8-(2- chloro-pyridin-3- yl)-9- cyclopropylmethyl- 1-methyl-9H- pyrido[3,4- b]indole LC6  1.11 382.1 [M + H].sup.+ 285 [00320]embedded image 6-Chloro-9- cyclopropylmethyl- 8-(2,6-dichloro- pyridin-3-yl)-1- methyl-9H- pyrido[3,4- b]indole LC6  1.17 416.1 [M + H].sup.+ 286 [00321]embedded image [5-(6-Chloro-9- cyclopropylmethyl- 1-methyl-9H- pyrido[3,4-b]indol- 8-yl)-pyrimidin-2- yl]-dimethyl-amine LC8  3.45 392.2 [M + H].sup.+ 287 [00322]embedded image 6-Chloro-9- cyclopropylmethyl- 8-(1-methyl-1H- pyrazol-4-yl)-9H- pyrido[3,4- b]indole LC3  1.03 337.1 [M + H].sup.+ 288 [00323]embedded image 6-Chloro-9- cyclopropylmethyl- 8-(1-pyridin-3- ylmethyl-1H- pyrazol-4-yl)-9H- pyrido[3,4- b]indole LC4  1.46 414.2 [M + H].sup.+ 289 [00324]embedded image 6-Chloro-9- cyclopropyl-8-(4- methoxy-phenyl)- 9H-pyrido[3,4- b]indole LC4  1.59 349.2 [M + H].sup.+ 290 [00325]embedded image 6-Chloro-9- cyclopropylmethyl- 1-methyl-8-(1- pyridin-3- ylmethyl-1H- pyrazol-4-yl)-9H- pyrido[3,4- b]indole LC4  1.46 428.3 [M + H].sup.+ 291 [00326]embedded image 6-Chloro-8-(4- chloro-phenyl)-9- cyclopropylmethyl- 9H-pyrido[3,4- b]indole LC4  1.75 367.3 [M + H].sup.+ 292 [00327]embedded image 2-(6-Chloro-9- cyclopropylmethyl- 1-methyl-9H- pyrido[3,4-b]indol- 8-yl)-benzamide LC4  1.52 390.2 [M + H].sup.+ 293 [00328]embedded image 6-Chloro-9- cyclopropylmethyl- 8-(4-methoxy- phenyl)-1-methyl- 9H-pyrido[3,4- b]indole LC8  3.88 377.2 [M + H].sup.+ 294 [00329]embedded image [6-Chloro-5-(6- chloro-9- cyclopropylmethyl- 9H-pyrido[3,4- b]indol-8-yl)- pyridin-2-yl]- dimethyl-amine LC4  1.85 411.1 [M + H].sup.+

    [0560] (a) Isolated in the form of 6-chloro-1-methyl-8-(4-phenoxy-phenyl)-9H-pyrido[3,4-b]indole hydrochloride

    [0561] Exemplary .sup.1H NMR data of example compounds

    Example 6

    [0562] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=8.45 (d, 1H), 8.41 (dd, 1H), 8.27 (d, 1H), 8.10 (d, 1H), 8.06 (d, 1H), 7.42-7.46 (m, 2H), 7.25 (dd, 1H), 6.68 (d, 1H), 5.52 (s, 2H), 4.29 (q, 2H), 2.89 (s, 3H), 2.55 (s, 3H), 0.68 (t, 3H)

    Example 7

    [0563] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=8.54 (d, 1H), 8.31 (d, 1H), 8.17 (t, 1H), 8.13 (d, 1H), 7.74 (d, 1H), 7.27 (d, 1H), 6.88 (d, 2H), 6.69 (d, 1H), 5.16-5.28 (m, 1H), 5.04-5.15 (m, 1H), 4.13 (br s, 1H), 3.37 (br s, 1), 2.85 (s, 3H), 2.06 (s, 3H), 0.80 (t, 3H)

    Example 8

    [0564] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=11.14 (s, 1H), 8.26 (d, 1H), 8.01 (d, 1H), 7.54-7.75 (m, 2H), 7.46 (s, 1H), 7.16-7.21 (m, 2H), 4.53 (d, 2H), 4.36 (d, 2H), 4.16 (s, 2H), 2.90 (s, 3H), 2.79 (s, 3H), 1.41 (s, 3H)

    Example 10

    [0565] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=9.22 (s, 1H), 8.48-8.53 (m, 2H), 8.28 (d, 1H), 8.04 (s, 1H), 7.71 (s, 1H), 7.42 (d, 1H), 5.21 (q, 2H), 4.95 (t, 1H), 4.26 (t, 2H), 3.80 (q, 2H)

    Example 13

    [0566] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=11.17 (s, 1H), 8.31 (d, 1H), 8.23 (d, 1H), 8.01 (d, 1H), 7.84 (d, 1H), 7.62-7.68 (m, 2H), 7.49 (d, 1H), 7.43 (d, 1H), 7.10-7.15 (m, 2H), 6.28 (t, 1H), 4.56 (t, 2H), 4.44 (t, 2H), 2.77 (s, 3H)

    Example 21

    [0567] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=15.26 (br s, 1H), 8.74 (d, 1H), 8.59 (s, 1H), 8.11 (s, 1H), 7.76 (s, 1H), 7.67 (d, 1H), 4.41 (q, 2H), 3.96 (s, 3H), 3.18 (s, 3H), 2.80 (s, 3H), 0.89 (t, 3H)

    Example 22

    [0568] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=8.61-8.73 (m, 2H), 8.51 (br d, 1H), 7.58 (d, 1H), 7.49-7.54 (m, 2H), 7.10-7.15 (m, 2H), 3.86 (s, 3H), 3.68 (s, 3H), 3.14 (s, 3H)

    Example 27

    [0569] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=12.31 (br s, 1H), 8.57 (d, 1H), 8.48 (d, 1H), 7.60-7.74 (m, 3H), 7.13-7.21 (m, 2H), 3.87 (s, 3H), 3.05 (s, 3H), 2.98 (s, 3H)

    Example 33

    [0570] .sup.1H NMR (600 MHz, DMSO-d.sub.6): δ (ppm)=12.66 (br s, 1H), 9.11 (s, 1H), 8.70 (br d, 1H), 8.59 (d, 1H), 8.01 (d, 1H), 7.68-7.73 (m, 2H), 7.18-7.21 (m, 2H), 3.88 (s, 3H), 3.06 (s, 3H)

    Example 39

    [0571] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=11.18 (s, 1H), 8.31 (d, 1H), 8.24 (d, 1H), 8.01 (d, 1H), 7.68 (m, 2H), 7.44 (d, 1H), 7.27-7.33 (m, 2H), 7.22 (s, 1H), 6.91 (s, 1H), 5.25 (s, 2H), 3.73 (s, 3H), 2.78 (s, 3H)

    Example 45

    [0572] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=11.11 (br s, 1H), 8.25 (d, 1H), 8.00 (d, 1H), 7.83 (d, 1H), 7.58-7.64 (m, 2H), 7.48 (d, 1H), 7.44 (s, 1H), 7.08-7.14 (m, 2H), 6.27 (t, 1H), 4.56 (t, 2H), 4.44 (t, 2H), 2.89 (s, 3H), 2.78 (s, 3H)

    Example 46

    [0573] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=9.11 (s, 1H), 8.61 (d, 1H), 8.46 (d, 1H), 8.32 (d, 1H), 8.27 (d, 1H), 7.82 (d, 1H), 7.54 (d, 1H), 4.07 (dd, 1H), 3.76 (dd, 1H), 0.69-0.80 (m, 1H), −0.17-0.28 (m, 2H), 0.03-0.16 (m, 2H)

    Example 55

    [0574] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ (ppm)=9.47 (s, 1H), 8.85 (d, 1H), 8.77 (d, 1H), 8.69 (br d, 1H), 833 (d, 1H), 7.84 (d, 1H), 7.76 (d, 1H), 4.18-4.36 (m, 1H), 3.74-3.95 (m, 1H), 0.99 (t, 3H)

    Example 116

    [0575] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=8.58 (d, 1H), 8.54 (dd, 1H), 8.40 (d, 1H), 8.29 (s, 1H), 8.28 (d, 1H), 8.10 (d, 1H), 7.82 (s, 1H), 7.69-7.74 (m, 1H), 7.40-7.46 (m, 1H), 7.38 (d, 1H), 5.51 (s, 2H), 4.34 (q, 2H), 2.91 (s, 3H), 0.74 (t, 3H)

    Example 148

    [0576] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=8.81 (d, 1H), 8.77 (d, 1H), 8.60 (br d, 1H), 7.62-7.68 (m, 4H), 7.61 (d, 1H), 4.18 (q, 2H), 3.12 (s, 3H), 0.86 (t, 3H)

    Example 149

    [0577] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ (ppm)=12.45 (s, 1H), 8.71 (d, 1H), 8.65 (d, 1H), 8.54 (br d, 1H), 7.66-7.72 (m, 3H), 7.15-7.22 (m, 2H), 3.88 (s, 3H), 3.06 (s, 3H)

    Example 184

    [0578] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ (ppm)=11.18 (s, 1H), 8.29 (d, 1H), 8.23 (d, 1H), 8.00 (d, 1H), 7.40-7.45 (m, 3H), 6.93 (d, 1H), 4.22 (t, 2H), 2.87 (t, 2H), 2.78 (s, 3H), 1.96-2.03 (m, 2H)

    Example 209

    [0579] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=8.87 (d, 1H), 8.76 (d, 1H), 8.57 (d, 1H), 8.11 (s, 1H), 7.76 (d, 1H), 7.68 (d, 1H), 4.45 (q, 2H), 3.97 (s, 3H), 3.15 (s, 3H), 0.92 (t, 3H)

    Example 215

    [0580] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=11.30 (s, 1H), 8.40 (d, 1H), 8.25 (d, 1H), 8.08 (d, 1H), 8.03 (d, 1H), 7.48 (d, 1H), 6.75-6.84 (m, 2H), 6.10 (s, 2H), 2.79 (s, 3H)

    Example 228

    [0581] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=11.18 (s, 1H), 8.31 (d, 1H), 8.24 (d, 1H), 8.01 (d, 1H), 7.63-7.68 (m, 2H), 7.44 (d, 1H), 7.12-7.16 (m, 2H), 3.92-4.00 (m, 2H), 2.78 (s, 3H), 2.53-2.67 (m, 3H), 2.32-2.44 (m, 2H), 2.26 (s, 3H), 1.93-2.03 (m, 1H), 1.49-1.58 (m, 1H)

    Example 256

    [0582] .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ (ppm)=8.66-8.78 (m, 2H), 8.54 (d, 1H), 7.46-7.59 (m, 3H), 7.14-7.21 (m, 2H), 4.54 (d, 2H), 4.35 (d, 2H), 4.33 (q, 2H), 4.16 (s, 2H), 3.10 (s, 3H), 1.42 (s, 3H), 0.84 (t, 3H)

    Biologic Examples

    [0583] A) Chondrogenesis Activity Assay in ATDC5 Cells

    [0584] The chondrogenic potential of the compounds of the invention was determined using clonal mouse chondrogenic ATDC5 cells (T. Atsumi et al., Cell Differ. Dev. 1990, 30, 109-116). ATDC5 cells are derived from mouse embryonic AT805 teratocarcinoma cells and are used frequently to study the multistep chondrogenic differentiation process from precursor cells into chondrocytes (C. Shukunami et al., Exp. Cell Res. 1998, 241, 1-11; H. Akiyami et al., J. Bone Miner. Res. 1996, 11, 22-28; H. Akiyami et al., Biochem. Biophys. Res. Commun. 1997, 235, 142-147; C. Shukunami et al., J. Cell Biol. 1996, 133, 457-468; C. Shukunami et al., J. Bone Miner. Res. 1997, 12, 1174-1188). Undifferentiated ATDC5 cells grow in vitro until confluence, showing a fibroblast-like morphology. In the presence of insulin, cells undergo transient condensation and form numerous nodular structures (cartilage nodules). The cartilagenous nature of these nodules was shown by Alcian Blue staining as evidence of the production of proteoglycan (aggrecan) and collagen type II expression (by expression analysis), both molecular markers of chondrocytes (C. Shukunami et al., J. Cell Biol. 1996, 133, 457-468).

    [0585] Chondrogenic differentiation of ATDC5 cells into chondrocytes by the compounds of the invention was determined by measuring the induction of type II collagen protein as a marker of chondrocytes, a structural component of the extracellular matrix which constitutes more than 80% of cartilage mass (D. R. Eyre, Clin. Orthop. Relat. Res. 2004, 427 Suppl, S118-S122). ATDC5 cells were obtained from RIKEN and cultured as monolayer in basal culture medium (Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F12, Invitrogen, #31331-093) supplemented with 10 μg/ml Transferrin (Roche, #10652 202001), 3×10E-8 M sodium selenite (Sigma, #S-5261) and 5% Fetal Calf Serum (FCS Gold, PAA, #A15-251)), at 37° C. and 5% CO.sub.2 in plastic flasks of 75 cm.sup.2 or 300 cm.sup.2 subconfluently for propagation. ATDC5 cells were harvested and resuspended in differentiation culture medium that consisted of basal culture medium complemented with 10 μg/ml insulin (Sigma, #19278), to initiate chondrogenic differentiation. For studying the effect of test compounds, the ATDC5 cells were plated in 96-well plates (3.0×10E4 cells per well in 200 μl of differentiation culture medium). Test compounds dissolved in DMSO were added to yield increasing compound concentrations, typically from 40 nM to 10 μM, and a final DMSO concentration of 0.1%. On each 96-well plate, control wells containing no test compound but the same concentration of DMSO, i.e. untreated cells, were included and used as reference for the determination of the effect of the test compounds on the cells, and wells containing an internal reference compound at a concentration of 10 μM were included and used as positive control and for standardizing the results. Cells in the wells were incubated for 4 days at 37° C. and 5% CO.sub.2, followed by quantification of intracellular collagen type II.

    [0586] Quantification of collagen type II in the ATDC5-cells was done by immunofluorescence staining of collagen type II, and fluorescence intensity was measured using a cellular high content imaging system (ImageXpress, Molecular Device). For immunofluorescence staining of intracellular collagen, medium was removed after compound incubation and cells were fixed with 200 μl/well methanol/water (95%/5%). Cells were permeabilized in 200 μl/well phosphate-buffered saline (PBS) containing 0.2% Triton X100 for 15 min and, after removal of the solution, blocked for 30 min with 200 μl/well PBS/0.2% Triton X100 containing 1% bovine serum albumin (BSA) to avoid unspecific binding. After the blocking step, the solution was removed and collagen type II staining solution was added. A primary antibody mouse anti-collagen type II solution (Quartett Immundiagnostika, #031502302) was diluted 1:100 in PBS/1% BSA, and 50 μl of the solution was added to each well and incubated for 1 hour at room temperature. After washing three times with PBS, 50 μl per well of a second antibody solution was added and incubated for 1 hour. The second antibody solution contained PBS/1% BSA with a 1:250 dilution of goat anti-mouse IgG (H+L) antibody coupled with fluorescent dye Alexa Fluor 488 (Invitrogen, #A11029), and fluorescent dye Hoe 33342 for staining nuclei with a final concentration of 2 μg/ml. Fluorescence signal intensity was measured for fluorescent dye Alexa Fluor 488 and for dye Hoe 33342, and the signals integrated over 9 fields within each well of a 96-well plate.

    [0587] The change of the collagen type II-derived fluorescence signal intensity was calculated for each compound concentration relative to the control (untreated cells, i.e. no test compound added), and an EC.sub.50 value (effective concentration 50 (in μM (micromol/liter)), i.e. the compound concentration at which the effect of the compound on collagen type II induction reaches 50% of the maximum induction) was calculated using a sigmoidal signal fit procedure. To allow the comparison of compound activities determined in different experiments, given the natural biological variation of the chondrogenic response between different experiments, an internal reference compound at a concentration of 10 μM was included in all experiments, and for each concentration collagen type II induction was calculated in percent in relation to the internal reference compound at 10 μM. The maximum percent induction (relative to the internal reference compound at a concentration of 10 μM) of a compound is termed E.sub.max. EC.sub.50 values (in μM) and E.sub.max values (in percent) obtained for compounds of the invention in this test are given in Table 2. In Table 2 the E.sub.max value “a” denotes a maximum percent induction of less than 20%, the E.sub.max value “b” denotes a maximum percent induction from 20% to less than 50%, the E.sub.max value “c” denotes a maximum percent induction from 50% to less than 80%, and the E.sub.max value “d” denotes a maximum percent induction of more than 80%, in each case relative to the internal reference compound at a concentration of 10 μM.

    TABLE-US-00002 TABLE 2 Example no. EC.sub.50 [μM] E.sub.max 1 1.7 a 2 0.71 d 3 >10 b 4 0.83 d 5 0.49 d 6 2.2 c 7 >3.3 a 8 0.12 d 9 1.4 d 10 0.69 d 11 3.9 c 12 >10 b 13 0.12 d 14 0.86 d 15 1.0 c 16 2.8 c 17 3.6 d 18 1.0 d 19 >10 c 20 3.9 c 21 >3.3 b 22 0.63 d 23 1.5 c 24 2.0 c 25 0.17 d 26 0.53 c 27 0.097 d 28 0.19 d 29 >1.1 a 30 0.27 d 31 0.25 d 32 0.37 c 33 0.12 d 34 >1.1 b 35 3.1 d 36 0.10 d 37 >3.3 b 38 2.6 c 39 0.17 d 40 0.15 d 41 0.24 c 42 0.51 c 43 >6.6 a 44 0.67 d 45 0.13 d 46 1.4 d 47 >10 b 48 0.041 a 49 0.041 a 50 0.13 a 51 >10 a 52 1.2 c 53 2.1 c 54 1.1 b 55 0.65 d 56 0.46 b 57 1.6 a 58 1.5 b 59 1.3 d 60 2.5 b 61 5.6 d 62 1.2 d 63 0.59 b 64 3.9 d 65 2.3 b 66 2.1 c 67 0.74 b 68 7.6 c 69 2.0 c 70 0.57 d 71 1.5 a 72 5.8 b 73 0.44 b 74 0.92 b 75 1.5 b 76 >10 b 77 0.58 d 78 >10 a 79 3.9 d 80 1.2 b 81 4.1 d 82 2 b 83 2.1 b 84 >10 b 85 >10 b 86 0.75 d 87 1.5 c 88 1.7 a 89 >10 d 90 >10 a 91 1.5 d 92 2.5 d 93 2.2 d 94 0.75 d 95 0.58 c 96 0.85 c 97 >1.1 a 98 2.4 d 99 >10 a 100 1.0 a 101 8.2 b 102 2.1 b 103 1.9 d 104 3.5 d 105 1.0 c 106 >10 b 107 0.56 b 108 1.4 d 109 0.72 c 110 >10 b 111 1.8 c 112 0.49 d 113 0.78 d 114 5.7 b 115 0.49 d 116 0.39 d 117 >10 a 118 >1.1 b 119 >3.3 a 120 1.4 c 121 0.59 d 122 >3.3 b 123 0.26 b 124 2.6 a 125 1.0 d 126 >10 b 127 0.38 c 128 3.9 a 129 0.86 d 130 0.38 d 131 0.78 d 132 4.5 b 133 >10 b 134 >10 a 135 0.33 c 136 4.1 c 137 1.4 b 138 >10 a 139 >10 a 140 0.91 b 141 >10 a 142 0.52 d 143 0.97 b 144 0.58 a 145 0.48 a 146 0.041 a 147 0.84 d 148 0.046 d 149 0.33 d 150 0.51 d 151 0.54 c 152 0.35 a 153 4.5 c 154 8.8 b 155 >3.3 a 156 1.5 a 157 >10 b 158 1.9 d 159 1.3 c 160 >10 a 161 2.0 d 162 0.59 d 163 0.69 d 164 0.23 d 165 >10 a 166 0.84 d 167 >10 b 168 0.99 d 169 1.1 d 170 >10 b 171 1.9 d 172 > 10 b 173 1.9 d 174 7.3 c 175 >10 a 176 > 10 b 177 >10 b 178 >10 b 179 >10 d 180 0.33 d 181 2.1 d 182 0.22 d 183 2.3 c 184 3.0 c 185 >10 b 186 0.10 d 187 2.2 d 188 0.64 d 189 0.50 d 190 0.72 d 191 0.55 d 192 0.64 d 193 >10 b 194 0.14 d 195 >10 a 196 0.80 c 197 0.63 d 198 1.8 b 199 > 10 a 200 >10 b 201 0.60 d 202 0.64 d 203 >10 a 204 0.19 c 205 3.3 d 206 >10 b 207 1.6 d 208 1.9 d 209 0.28 d 210 >3.3 a 211 >3.3 a 212 >3.3 a 213 >1.1 a 214 > 3.3 a 215 0.23 d 216 >10 a 217 >3.3 b 218 >1.1 a 219 >3.3 a 220 >10 b 221 > 3.3 b 222 >10 a 223 >3.3 b 224 0.31 d 225 1.7 d 226 0.19 c 227 0.33 d 228 0.15 c 229 0.28 c 230 > 3.3 a 231 >10 b 232 0.33 d 233 >10 b 234 5.8 c 235 >3.3 b 236 >3.3 a 237 0.50 d 238 1.4 d 239 0.25 c 240 0.49 c 241 >1.1 b 242 0.57 d 243 0.66 d 244 1.1 d 245 >1.1 a 246 2.9 c 247 >3.3 a 248 >3.3 b 249 > 3.3 b 250 >3.3 c 251 >10 a 252 1.6 d 253 >10 b 254 > 3.3 a 255 > 3.3 a 256 0.16 d 257 0.31 d 258 >3.3 a 259 0.43 d 260 >1.1 a 261 1.5 d 262 2.3 d 263 0.55 d 264 >1.1 b 265 >1.1 a 266 0.11 d 267 > 3.3 c 268 0.34 d 269 >3.3 b 270 >1.1 b 271 > 3.3 b 272 1.9 d 273 0.17 a 274 >3.3 a 275 0.30 d 276 0.42 d 277 > 3.3 a 278 >3.3 b 279 1.8 b 280 >10 a 281 0.11 a 282 3.6 a 283 0.41 a 284 4.2 a 285 6.1 b 286 0.041 a 287 0.23 c 288 0.52 d 289 0.75 d 290 >10 b 291 1.7 c 292 >6.6 a 293 4.5 c 294 1.2 d

    [0588] B) Chondrogenesis Activity Assay in Primary Human Chondrocyte Pellet Cultures

    [0589] In this assay human articular chondrocytes are harvested by enzymatic digestion from articular cartilage and passaged several times to dedifferentiate the chondrocytes and to propagate the cells. Cells are cultured as cell pellets in the presence of the compounds of the invention over 2 weeks, and the chondrogenic differentiation is quantified by the production of the chondrogenic marker aggrecan (proteoglycan).

    [0590] In detail, primary chondrocytes were harvested by enzymatic digestion from cartilage of osteoarthritis patients undergoing knee joint replacement surgery, and cultured in vitro. Cells were passaged one or two times and aliquots were cryopreserved. To initiate chondrogenesis experiments, cell aliquots were thawn, cultured in Chondrocyte Growth Medium (CGM, Lonza, #CC-3216) and passaged twice to further propagate and dedifferentiate cells. Pellet cultures were initiated by seeding 2.5×10E5 cells per well into a 96-well deep well plate in 600 μl chondrocyte differentiation medium consisting of Dulbecco's Modified Eagle Medium (DMEM, Invitrogen, #41966), 1×ITS-solution (Insulin/Transferrin/sodium selenite; 100× solution: Invitrogen, #51500056), 5 μg/mL linoleic acid (Sigma, #L1012-1G), 1× nonessential amino acids (100× solution: Invitrogen, #11140); 10 nM Dexamethasone, 2 ng/ml TGF-β1 (transforming growth factor β1) and 10 μg/ml ascorbic acid. Cells were spun to pellets by centrifugation (10 min, 400×g). Test compounds dissolved in DMSO were added to yield increasing compound concentrations, typically from 40 nM to 10 μM, and a final DMSO concentration of 0.1%, and the cell pellets cultured at 37° C. and 5% CO.sub.2 for 2 weeks. On each 96-well plate, control wells containing no test compound but the same concentration of DMSO, i.e. untreated cells, were included and used as reference for the determination of the effect of the test compounds on the cells, and wells containing an internal reference compound at a concentration of 10 μM were included and used as positive control and for standardizing the results. Chondrocyte differentiation medium and the compounds were exchanged twice weekly until harvest of the pellets.

    [0591] For harvest of the pellets, medium was removed and the pellet was homogenized by enzymatic digestion. 100 μl of protease solution containing 0.4 mg/ml papain, 50 mM sodium phosphate, 4 mM EDTA and 0.48 mg/ml L-cysteine was added to the pellet in each well of a 96-well plate, the plate was sealed with a plate sealer foil (SILVERseal, Greiner Bio-One) and incubated for 4 to 6 hours at 65° C. with agitation. The concentration of proteoglycan (aggrecan) was determined by quantification of the sulfated glycosaminoglycan side chains of aggrecan using the Blyscan assay (Kit from Biocolor). Aliquots of the papain-digest were transferred to 400 μl of Blyscan dye solution and incubated at room temperature for 45 min with agitation (1200 rpm). The mixture was centrifuged at 3760×g for 45 min, the supernatant was discarded, and the stained pellet was redissolved in 400 μl of Blyscan dissociation reagent by rotation until the precipitate completely resolved. Optical density was measured at 656 nm using a Tecan Saphire2 instrument, and the amount of proteoglycan determined against a standard curve with chondroitin-4-sulfate.

    [0592] Induction of proteoglycan (aggrecan) was calculated for each compound concentration relative to the control (untreated cells, i.e. no test compound added), and an EC.sub.50 value (effective concentration 50 (in μM (micromol/liter)), i.e. the compound concentration at which the effect of the compound on proteoglycan (aggrecan) induction reaches 50% of the maximum induction) was calculated using a sigmoidal signal fit procedure. To allow the comparison of compound activities determined in different experiments, given the natural biological variation of the chondrogenic response between different experiments, an internal reference compound at a concentration of 10 μM was included in all experiments, and for each concentration proteoglycan (aggrecan) induction was calculated in percent in relation to the internal reference compound at 10 μM. The maximum percent induction (relative to the internal reference compound at a concentration of 10 μM) of a compound is termed E.sub.max. EC.sub.50 values (in μM) and E.sub.max values (in percent) obtained for compounds of the invention in this test are given in Table 3. In Table 3 the E.sub.max value “a” denotes a maximum percent induction of less than 40%, the E.sub.max value “b” denotes a maximum percent induction from 40% to less than 100%, the E.sub.max value “c” denotes a maximum percent induction from 100% to less than 200%, and the E.sub.max value “d” denotes a maximum percent induction of more than 200%, in each case relative to the internal reference compound at a concentration of 10 μM.

    TABLE-US-00003 TABLE 3 Example no. EC.sub.50 [μM] E.sub.max 2 4.4 d 5 1.3 d 8 0.26 c 10 2.4 b 11 7.9 b 13 1.1 d 19 6.9 b 21 4.3 d 22 4.0 d 27 3.5 d 29 1.5 c 33 0.83 c 34 3.8 d 37 4.3 c 39 3.7 d 45 0.22 b 46 1.0 c 55 0.95 c 97 5.1 d 116 1.3 c 117 27 a 118 1.3 c 119 21 b 122 3.6 c 149 1.1 c 167 13 a 171 4.0 d 184 1.9 b 194 0.83 c 195 14 a 199 22 a 200 4.4 c 206 19 a 209 2.6 d 212 8.9 a 213 1.2 c 214 1.3 c 215 0.99 d 217 4.9 d 218 40 a 223 27 a 228 0.14 c 230 21 a 235 5.1 d 236 4.6 d 241 1.5 c 244 4.9 d 245 10 b 247 3.8 d 250 4.9 c 251 9.1 b 256 1.3 d 258 8.2 b 264 1.5 b 265 3.8 a 270 4.3 b 271 4.9 d 288 1.2 d

    [0593] The compounds of the invention can also be tested in the animal models described in biological examples C) and D), which are in vivo models of osteoarthritis (OA) in rodents.

    [0594] C) Joint Instability Induced OA in Rats after Anterior Cruciate Ligament Transection and Partial Meniscectomy (ACLT-pMx)

    [0595] In this model osteoarthritis is induced via ACLT-pMx surgery in rats and assessment of histopathological joint damage is conducted as primary readout. Under general anesthesia by isoflurane (4%-5% in 3 L/min O.sub.2) the right leg of the rats is shaved and disinfected with Cutasept® (Beiersdorf, Germany). Then, with the leg in extension, a para-patellar skin incision is made on the medial side of the joint. After dislocating the patella laterally, an incision of the joint capsule on the medial side of the patellar tendon is made to access the joint space. The anterior cruciate ligament is transected using a modified sharpened hook (“Ohrhebel nach Wagener”; Aesculap, #OF 285 R). Then the medial meniscus is gently retracted and the cranial part of the meniscus (30%) carefully excised by using an Aesculap microscalpel to ensure that the cartilage of the femur and the tibia is not damaged. During the surgery the joint space is lavaged with 0.9% sterile saline to remove all blood from the joint and to prevent damage by drying of the tissue. After repositioning of the patella the joint capsule is closed with Safil® absorbable sutures (B. Braun Melsungen, Germany). The skin is closed with Dafilon® 3/0 sutures (B. Braun Melsungen, Germany). Buprenorphine hydrochloride is given subcutaneously (0.06 mg/kg) as a post-surgical analgesic treatment.

    [0596] Treatment onset with a test compound is seven days after surgery. The animals receive intra-articular injections of 0.1 to 1 mg/joint of the test compound suspended in 50 μl of vehicle into the operated knee joint in up to weekly intervals, whereas the control animals receive injections of 50 μl of the vehicle. At day 28 post surgery all animals are sacrificed for histology and histopathological analysis, which is performed as described in biological example D).

    [0597] D) Spontaneous Model of OA in Dunkin Hartley Guinea Pigs

    [0598] In this model Dunkin Hartley guinea pigs of strain HsdDhl:DH (Harlan Laboratories, The Netherlands), which is a widely used strain for spontaneous animal models of OA since their histological and biochemical changes resemble that of human OA (A. M. Bendele et al., Arthritis Rheum. 1988, 31, 561-565), are used at the age of 6 months. It is known that histological changes start at the age of about 3 months and disease severity increases with age (P. A. Jimenez et al., Lab. Anim. Sci. 1997, 47, 598-601). Therefore treatment onset in this model is at the age of 6 months and continued until animals reach an age of a minimum of 12 months.

    [0599] The animals receive intra-articular injections of 0.1 to 3 mg/joint of the test compound suspended in 100 μl of vehicle into the right knee joint in up to weekly intervals, whereas the control animals receive injections of 100 μl of the vehicle. After a minimum of 6 months treatment all animals are sacrificed for histology and histopathological analysis.

    [0600] Evaluation of the tests described in examples C) and D) is done in the following way.

    [0601] For histological processing of the tissue the right knee joints of the animals are excised at the mid-shaft of femur and tibia and placed in 10% formalin for 3 days, to fix the tissue. After fixing the knees are decalcified in formic acid (Immunocal®, Decal Chemical Corp., NY, USA) for 11 days, dehydrated in the Tissue Processor TP 1020@ (Leica, Germany) and embedded in paraffin. The paraffin-embedded complete knee is serially sectioned (coronal sections) on a rotary microtome at a thickness of 7 μm and the sections are stained with Hematoxylin/Eosin (H&E) or Safranin O/Fast Green (SO). For each knee 4 subregions of the knee joint (medial or lateral tibia or femur) are defined. From each subregion 5 sections with the most severe damage are selected and evaluated by two observers blinded to the treatment by using a modified Mankin score.

    [0602] Digital images from histological H&E-stained as well as SO-stained coronal sections of the whole knee joint are taken using a Zeiss AxioScanner®. After conversion into tif files and transfer into the digital image analysis software Visiopharm Integrator System (VIS; Version Nr. 3.0.15.0; Visiopharm, Denmark), the cartilage tissue, the chondrocytes, the SO-stained cartilage area and the subchondral bone are segmented. As region of interest (ROI) a rectangle of 1.2×0.5 mm covering the most affected area of cartilage and the underlying subchondral bone in the medialtibial plateau are chosen. The degree of cartilage destruction and subchondral bone sclerosis is then quantified by measuring the following parameters: fibrillation index (FI; the width of the region of interest (box) divided by the cartilage surface curvature length, i.e. measures of cartilage surface irregularity); cartilage area; chondrocyte number (cell number per residual cartilage area); absolute number of residual chondrocytes; proteoglycan containing (SO-stained) cartilage area; subchondral solid bone area.

    [0603] For statistical analysis of the data, results are given as median, inter-quartile and complete data range (histopathological scoring) or mean±SEM (histomorphometry). The statistical significance of the effect of a compound on the histomorphometrically assessed joint pathology is determined by a one way analysis of variance followed by Dunnett's test for multiple comparisons versus the vehicle-treated group. Kruskal-Wallis test and multiple comparisons by Dunn's Test versus the vehicle-treated group are applied for the semi-quantitative histopathological scores. SAS® v 8.2 via Everstat software v 5.0 interface is used for the statistical analyses.