Compounds and compositions for targeting the TP53-Y220C mutation

Abstract

Formula (I): Compounds of TIFF2026500291000179.tif2032 and pharmaceutically acceptable salts thereof, and compositions thereof are provided, which are useful for treating a variety of conditions associated with activation of p53.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 432,074, filed December 13, 2022, the entire disclosure of which is incorporated herein by reference. [Background technology]

[0002] The transcription factor TP53 functions as a tumor suppressor and is inactivated via mutation in approximately 50% of all tumors. TP53 regulates many intracellular metabolic pathways, including DNA damage repair, apoptosis, and senescence. The Y220C mutation, a common TP53 missense variant, is associated with more than 100,000 new cancer cases annually worldwide, primarily breast and ovarian cancer. The Y220C mutation is known to cause a major conformational change in the TP53 protein, creating a new protein pocket that could potentially provide a small-molecule drug candidate. Crucially, the cavity created by the mutation is distant from the TP53 surface involved in DNA recognition or protein-protein interactions, enabling the development of targeted chemical agents that stabilize the DNA-binding domain without inhibiting binding to its natural substrates. Stabilization allows for the restoration of TP53 function, thereby reactivating the TP53 tumor suppressor pathway and shrinking or eliminating tumors. Summary of the Invention [Means for solving the problem]

[0003] Provided herein are compounds of formula I:

[0004] [ka]

[0005] (In the formula, R 1 , R 2 , R 3 , R4 , X and p are as described herein. and pharmaceutically acceptable salts thereof, and compositions thereof. In one embodiment, the disclosed compounds of Formula I and pharmaceutically acceptable salts thereof act as covalent modifiers of TP53 Y220C. In one embodiment, the disclosed compounds of Formula I and pharmaceutically acceptable salts thereof stabilize TP53 Y220C. In one embodiment, the disclosed compounds of Formula I and pharmaceutically acceptable salts thereof restore wild-type tumor suppressor protein TP53 (WT TP53) function. In one embodiment, it is believed that the disclosed compounds are covalent modifiers of TP53 Y220C, which leads to stabilization of the mutant protein and can restore wild-type function to dysfunctional mutant proteins. See, e.g., the Exemplification section below.

[0006] Also included are pharmaceutical compositions containing the compounds and pharmaceutically acceptable salts of the disclosed compounds of Formula I, as well as methods for their preparation. DETAILED DESCRIPTION OF THE INVENTION

[0007] 1. General description of the compound In a first embodiment, provided herein is a compound of formula I:

[0008] [ka]

[0009] [In the formula, R 1 is selected from optionally substituted alkyl; R 2 is hydrogen, halo, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted heterocyclyl, -NR a R b , -ORc , -NHC(O)R c , -C(O)NR d R e , -C(O)R f and -SR g is selected from R 3 is hydrogen, halo, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted heterocyclyl, -NR a1 R b1 , -OR c1 , -NHC(O)R c1 , -C(O)NR d1 R e1 , -C(O)R f1 and -SR g1 is selected from R 4 is halo, cyano, optionally substituted alkyl and optionally substituted alkoxy; X is selected from halo, —S(O)alkyl, and —S(O)alkyl; R a , R a1 , R b , R b1 , R c and R c1 are each independently selected from hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted aryl, and optionally substituted heterocyclyl; R d , R e and R g are each independently selected from hydrogen, optionally substituted alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted heterocyclyl, —C(O)R′, —C(O)OR′, —C(O)NR′R″, —S(O)R′, and —S(O)R′; or R d and Re together with the nitrogen atom to which they are attached form an optionally substituted heterocyclyl or an optionally substituted heteroaryl; R d1 , R e1 and R g1 are each independently hydrogen, optionally substituted alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted heterocyclyl, -C(O)R' 1 , -C(O)OR' 1 , -C(O)NR' 1 R'' 1 , -S(O)R' 1 and -S(O)R' 1 or R d1 and R e1 together with the nitrogen atom to which they are attached form an optionally substituted heterocyclyl or an optionally substituted heteroaryl; R f and R f1 are each independently selected from hydrogen, optionally substituted alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, and optionally substituted heterocyclyl; R', R' 1 , R'' and R'' 1 are each independently selected from hydrogen and optionally substituted (C1-C4) alkyl; p is 0, 1, or 2. is a compound of

[0010] 2.Definition When used in the context of describing a chemical group that may have multiple points of attachment, the hyphen (-) designates the point of attachment of that group to the variable where it is defined. For example, -NR a R b means that the point of attachment for that group is on the nitrogen atom.

[0011] The terms "halo" and "halogen" refer to an atom selected from fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br), and iodine (iodo, -I).

[0012] The term "alkyl," when used alone or as part of a larger moiety such as "haloalkyl," means a saturated, straight-chain or branched, monovalent hydrocarbon radical.

[0013]

[0009] "Alkoxy" means an alkyl group attached through an oxygen linking atom represented by -O-alkyl. For example, "(C1-C4)alkoxy" includes methoxy, ethoxy, propoxy, and butoxy.

[0014] The term "haloalkyl" includes mono-, poly-, and perhaloalkyl groups, where the halogens are independently selected from fluorine, chlorine, bromine, and iodine.

[0011] A "haloalkoxy" is a haloalkyl group that is attached to another moiety through an oxygen atom, for example, --OCHF2 or --OCF3.

[0015] The term oxo refers to the group =O. The term "heteroaryl," used alone or as part of a larger moiety, refers to a 5- to 12-membered aromatic group containing 1 to 4 heteroatoms selected from N, O, and S. Heteroaryl groups can be monocyclic or bicyclic. Monocyclic heteroaryls include, for example, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, triazinyl, tetrazinyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, and the like. Bicyclic heteroaryls include groups in which a monocyclic heteroaryl ring is fused to one or more aryl or heteroaryl rings. Non-limiting examples include indolyl, imidazopyridinyl, benzoxazolyl, benzoxodiazolyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, quinazolinyl, quinoxalinyl, pyrrolopyridinyl, pyrrolopyrimidinyl, pyrazolopyridinyl, thienopyridinyl, thienopyrimidinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. When specified, it will be understood that optional substituents on a heteroaryl group may be present on any substitutable position, and include, for example, the position at which the heteroaryl is attached.

[0016] The term "heterocyclyl" refers to a 5- to 12-membered saturated or partially unsaturated heterocyclic ring containing 1 to 4 heteroatoms independently selected from N, O, and S. It can be monocyclic, bicyclic (e.g., bridged, fused, or spiro bicyclic rings), or tricyclic. The heterocyclyl ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure. Examples of such saturated or partially unsaturated heterocyclic groups include, without limitation, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, pyrrolidinyl, pyridinonyl, pyrrolidonyl, piperidinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, morpholinyl, dihydrofuranyl, dihydropyranyl, dihydropyridinyl, tetrahydropyridinyl, dihydropyrimidinyl, oxetanyl, azetidinyl, and tetrahydropyrimidinyl. Heterocyclyl groups can be monocyclic or bicyclic. The term "heterocyclyl" also includes, for example, another unsaturated heterocyclic group or an unsaturated heterocyclic group fused to an aryl or heteroaryl ring, such as tetrahydronaphthyridine, indolinone, dihydropyrrolotriazole, imidazopyrimidinequinolinone, dioxaspirodecane. When specified, it is also understood that optional substituents on a heterocyclic group may be present at any substitutable position and include, for example, the position to which the heterocyclyl is attached (e.g., in the case of an optionally substituted heterocyclyl or an optionally substituted heterocyclyl).

[0017] The term "spiro" refers to two rings that share one ring atom (eg, carbon). The term "fused" refers to two rings that share two adjacent ring atoms. The term "bridged" refers to two rings which share three ring atoms with each other.

[0018] The term "cycloalkyl," as used alone or as part of a larger moiety, refers to a saturated cycloaliphatic monocyclic or bicyclic ring system having 3 to 10 carbon ring atoms, as described herein, unless otherwise specified. Monocyclic cycloalkyl groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, and cyclooctyl. When specified, it will be understood that any substituent on a cycloalkyl group or alicyclic group may be present on any substitutable position and includes, for example, the position at which the cycloalkyl group is attached.

[0019] The term "optionally substituted" means that one or more hydrogens of the specified moiety may be replaced with a suitable substituent. Unless otherwise specified, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, as valence allows. Optional substituents include cyano (-CN), halo, imino (=NH), nitro (-NO2), oxo (=O), -C(O)R i , -C(O)OR i , -C(O)NR ii R iii , -C(O)SR i , -C(NR i )NR ii R iii , -C(S)R i , -C(S)OR i , -C(S)NR ii R iii , -OR i , -OC(O)R i , -OC(O)OR i , -OC(O)NR ii R iii , -OC(O)SR i , -OC(NR i )NR ii R iii , -OC(S)R i , -OC(S)OR i , -OC(S)NR ii R iii, -OP(O)(OR ii ) OR iii , -OS(O)R i , -OS(O)2R i , -OS(O)NR ii R iii , -OS(O)2NR ii R iii , -NR ii R iii , -NR i C(O)R iv , -NR i C(O)OR iv , -NR i C(O)NR ii R iii , -NR a C(O)SR iv , -NR i C(NR iv )NR ii R iii , -NR i C(S)R iv , -NR i C(S)OR iv , -NR i C(S)NR ii R iii , -NR i S(O)R iv , -NR i S(O)2R iv , -NR i S(O)NR ii R iii , -NR i S(O)NR ii R iv , -SR i , -S(O)R i , -S(O)2R i , -S(O)NR ii R iv , -S(O)NR ii R iv , alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, and heterocyclyl, wherein said alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, and heterocyclyl may each optionally contain one or more, in one embodiment one, two, three, or four, substituents Qa each optionally further substituted, where each R i , R ii , R iii and R iv is independently alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, or heterocyclyl, each of which is substituted with one or more, in one embodiment one, two, three, or four, substituents Q a and optionally substituted; or R ii and R iii together with the N atom to which they are attached, one or more, in one embodiment one, two, three or four, substituents Q a wherein each Q a are independently cyano, halo, imino, nitro, oxo, alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, C 6~14 Aryl, heteroaryl, heterocyclyl, -C(O)R v , -C(O)OR v , -C(O)NR vi R vii , -C(O)SR v , -C(NR v )NR vi R vii , -C(S)R v , -C(S)OR v , -C(S)NR vi R vii , -OR v , -OC(O)R v , -OC(O)OR v , -OC(O)NR vi R vii , -OC(O)SR v , -OC(NR v )NR vi R vii , -OC(S)R v , -OC(S)OR v , -OC(S)NR vi R vii , -OP(O)(OR v ) OR vi , -OS(O)Rv , -OS(O)2R v , -OS(O)NR vi R vii , -OS(O)2NR v R vii , -NR vi R vii , -NR v C(O)R viii , -NR e C(O)OR vi , -NR v C(O)NR vi R vii , -NR v C(O)SR vi , -NR v C(NR viii )NR vi R vii , -NR v C(S)R viii , -NR v C(S)OR vi , -NR v C(S)NR vi R vii , -NR v S(O)R viii , -NR v S(O)2R viii , -NR v S(O)NR vi R vii , -NR v S(O)NR vi R vii , -SR v , -S(O)R v , -S(O)2R v , -S(O)NR vi R vii , and -S(O)NR vi R vii where each R v , R vi , R vii and R viii is independently alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heteroaryl, or heterocyclyl; or (iii) R vi and R viiitogether with the N atom to which they are attached form a heterocyclyl.

[0020] In certain embodiments, where specified, one or more hydrogen atoms on the disclosed compounds may be replaced with deuterium. Such deuterated compounds may have one or more improved pharmacokinetic or pharmacodynamic properties (e.g., longer half-life) compared to the equivalent "non-deuterated" compound.

[0021] One or more of the compounds described herein may exist in various tautomeric forms and are part of this disclosure. The term "tautomer" or "tautomeric" refers to two or more interconvertible compounds / substituents resulting from the formal migration of at least one hydrogen atom and at least one change in valence. All such isomeric forms of such compounds are expressly included. Thus, when a compound herein is represented by a structural formula or designated herein by a chemical name, all tautomeric forms that may exist for that compound are encompassed by that structural formula.

[0022] Compounds with one or more chiral centers can exist in various stereoisomeric forms. Stereoisomers are compounds that differ only in their spatial arrangement. Stereoisomers include all diastereomeric, enantiomeric, and epimeric forms, as well as racemates and mixtures thereof. "Geometric isomer" refers to isomers that differ in the orientation of substituents in relationship to a carbon-carbon double bond, a cycloalkyl ring, or a bridged bicyclic system. The atoms (other than H) on each side of a carbon-carbon double bond can be in the E (substituents on opposite sides of the carbon-carbon double bond) or Z (substituents pointing up the same side) configuration. "Cis" refers to substituents pointing up the same side of the ring, while "trans" refers to substituents pointing up on opposite sides of the ring.

[0023]

[0023] When the stereochemical configuration at a chiral center in a compound having one or more chiral centers is depicted in its chemical name (e.g., where the configuration is indicated in the chemical name by "R" or "S") or structure (e.g., where the configuration is indicated by a "wedge" bond), the enrichment of the depicted configuration relative to the opposite configuration is greater than 50%, greater than 60%, greater than 70%, greater than 80%, greater than 90%, greater than 99%, or greater than 99.9%. "Enrichment of the depicted configuration relative to the opposite configuration" is a mole percent and is determined by dividing the number of compounds with the depicted stereochemical configuration at the chiral center by the total number of all compounds with the same or opposite stereochemical configuration in the mixture.

[0024]

[0024] When geometric isomers are depicted by name or structure, the enrichment of the depicted isomer relative to the opposite isomer is greater than 50%, greater than 60%, greater than 70%, greater than 80%, greater than 90%, greater than 99%, or greater than 99.9%. "Enrichment of the depicted isomer relative to the opposite isomer" is a mole percent and is determined by dividing the number of compounds with the depicted geometric configuration by the total number of all compounds with the same or opposite geometric configuration in the mixture.

[0025]

[0025] When a disclosed compound is named or depicted by structure without indicating stereochemistry, it is understood that the name or structure encompasses one of the possible stereoisomers or geometric isomers, free from the others, or a mixture of the stereoisomers or geometric isomers encompassed.

[0026] The terms "subject" and "patient" may be used interchangeably and refer to a mammal in need of treatment, such as pets (e.g., dogs, cats, etc.), livestock (e.g., cows, pigs, horses, sheep, goats, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, etc.). Typically, the subject is a human in need of treatment.

[0027]

[0027] The terms "inhibit," "inhibition," or "inhibiting" include a reduction in the baseline activity of a biological activity or process.

[0028] As used herein, the terms "treatment," "treat," and "treating" refer to reversing, alleviating, delaying the onset of, or inhibiting the progression of, a disease or disorder described herein or one or more symptoms thereof. In some embodiments, treatment may be administered after one or more symptoms have developed, i.e., therapeutic treatment. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual (e.g., in light of a history of symptoms and / or exposure to a particular microorganism or other susceptibility factor) prior to the onset of symptoms, i.e., prophylactic treatment. Treatment may also be continued after symptoms have resolved, e.g., to delay their recurrence.

[0029]

[0029] The term "pharmaceutically acceptable carrier" refers to a non-toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants, or vehicles that can be used in the compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts, or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and wool fat.

[0030] For use in medicine, salts of the compounds described herein refer to non-toxic "pharmaceutically acceptable salts." Pharmaceutically acceptable salt forms include pharmaceutically acceptable acidic / anionic salts or basic / cationic salts. Suitable pharmaceutically acceptable acid addition salts of the compounds described herein include, for example, salts of inorganic acids (e.g., hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, and sulfuric acid) and salts of organic acids (e.g., acetic acid, benzenesulfonic acid, benzoic acid, methanesulfonic acid, and p-toluenesulfonic acid). Compounds of the present teachings having an acidic group, such as a carboxylic acid, can form pharmaceutically acceptable salts with pharmaceutically acceptable bases. Suitable pharmaceutically acceptable base salts include, for example, ammonium salts, alkali metal salts (e.g., sodium salts and potassium salts), and alkaline earth metal salts (e.g., magnesium salts and calcium salts). Compounds having a quaternary ammonium group also contain counterions such as chloride, bromide, iodide, acetate, perchlorate, and the like. Other examples of such salts include hydrochlorides, hydrobromides, sulfates, methanesulfonates, nitrates, benzoates, and salts with amino acids such as glutamic acid.

[0031]

[0031] The term "effective amount" or "therapeutically effective amount" refers to an amount of a compound described herein that will induce a desired or beneficial biological or medical response in a subject, for example, a dosage of between 0.01 mg / kg body weight / day and 100 mg / kg body weight / day.

[0032] 3.Compound In a second embodiment, p in the compound of Formula I, or a pharmaceutically acceptable salt thereof, is 0, wherein the remaining variables are as described above for Formula I.

[0033] In a third embodiment, X in the compound of Formula I, or a pharmaceutically acceptable salt thereof, is selected from chloro, bromo, fluoro, -SO2CH3, and -SOCH, where the remaining variables are as described above for Formula I or in the second embodiment. Alternatively, as part of the third embodiment, X in the compound of Formula I, or a pharmaceutically acceptable salt thereof, is selected from chloro, bromo, fluoro, and -SO2CH3, where the remaining variables are as described above for Formula I or in the second embodiment. In another alternative, as part of the third embodiment, X in the compound of Formula I, or a pharmaceutically acceptable salt thereof, is chloro, where the remaining variables are as described above for Formula I or in the second embodiment.

[0034] In a fourth embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof is 1 is selected from (C1-C4)alkyl, halo(C1-C4)alkyl, cyano(C1-C4)alkyl, and —(C1-C4)alkyl(C3-C6)cycloalkyl, where the remaining variables are as described above for Formula I, or as described in the second or third embodiment. Alternatively, as part of the fourth embodiment, R in a compound of Formula I or a pharmaceutically acceptable salt thereof 1 is selected from (C1-C4)alkyl and halo(C1-C4)alkyl, where the remaining variables are as described above for Formula I, or as described in the second or third embodiment. In another alternative, as part of the fourth embodiment, R in a compound of Formula I or a pharmaceutically acceptable salt thereof 1is selected from -CH2CH3, -CH2CF3, -CH2CF2CH3, -CH2CF2CF3, -CH2CN and -CH2-cyclopropyl, where the remaining variables are as described above for Formula I, or as described in the second or third embodiment. In another alternative, as part of the fourth embodiment, R in a compound of Formula I or a pharmaceutically acceptable salt thereof 1 is selected from -CH2CH3, -CH2CF3, -CH2CF2CH3 and -CH2CF2CF3, where the remaining variables are as described above for Formula I, or as described in the second or third embodiment. In yet another alternative, as part of the fourth embodiment, R in a compound of Formula I or a pharmaceutically acceptable salt thereof 1 is selected from -CH2CF3, where the remaining variables are as described above for Formula I, or as described in the second or third embodiment.

[0035] In a fifth embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof 2 is hydrogen, halo, (C1-C4)alkyl, halo(C1-C4)alkyl, (C3-C6)cycloalkyl, -(C1-C4)alkyl(C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, -(C1-C4)alkyl[5- to 7-membered heteroaryl], phenyl, -(C1-C4)alkylphenyl, 4- to 6-membered heterocyclyl, -(C1-C4)alkyl[4- to 6-membered heterocyclyl], -NR a R b , -OR c , -(C1-C4) alkyl OR c , -C(O)R f , -C(O)NR d R e , -(C1-C4) alkylNR d R e , -(C1-C4) alkylC(O)R f , -(C1-C4) alkylC(O)NR d R e and -SR gwherein each occurrence of (C-C)cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclyl is selected from R 5 and optionally substituted with 1 to 3 groups selected from (or R 2 is hydrogen, halo, (C1-C4)alkyl, halo(C1-C4)alkyl, (C3-C6)cycloalkyl, -(C1-C4)alkyl(C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, -(C1-C4)alkyl[5- to 7-membered heteroaryl], phenyl, -(C1-C4)alkylphenyl, 4- to 6-membered heterocyclyl, -(C1-C4)alkyl[4- to 6-membered heterocyclyl], -NR a R b , -OR c , -(C1-C4) alkyl OR c , -C(O)R f , -C(O)NR d R e , -(C1-C4) alkylNR d R e , -(C1-C4) alkylNR a C(O)R b , -(C1-C4) alkylC(O)R f , -(C1-C4) alkylC(O)NR d R e and -SR g wherein each occurrence of (C-C)cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclyl is selected from R 5 and optionally substituted with 1 to 3 groups selected from R a , R b and R care each independently selected from hydrogen, (C1-C4)alkyl, halo(C1-C4)alkyl, (C3-C6)cycloalkyl, -(C1-C4)alkyl(C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, -(C1-C4)alkyl[5- to 7-membered heteroaryl], phenyl, -(C1-C4)alkylphenyl, 4- to 6-membered heterocyclyl, -(C1-C4)alkyl[4- to 6-membered heterocyclyl], wherein at each occurrence of (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclyl, said groups each may be selected from R 6 and optionally substituted with 1 to 3 groups selected from R d , R e and R g are each independently hydrogen, (C1-C4) alkyl, halo(C1-C4) alkyl, -(C1-C4) alkyl OR', -(C1-C4) alkyl NR'R'', -(C1-C4) alkyl C(O)NR'R'', -(C1-C4) alkyl C(O)R', -(C1-C4) alkyl C(O)OR', -(C1-C4) alkyl S(O)R', -(C1-C4) alkyl S(O)2R', (C1-C4) alkyl phenyl, phenyl, (C3-C6) cycloalkyl and -S(O)R', wherein each occurrence of (C3-C6)cycloalkyl, phenyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclyl is selected from -(C1-C4)alkyl[5- to 7-membered heteroaryl], 5- to 7-membered heteroaryl, 4- to 6-membered heterocyclyl, -(C1-C4)alkyl[4- to 6-membered heterocyclyl], -C(O)R', -C(O)OR', -C(O)NR'R'', -S(O)R', and -S(O)R', 7 or R d and R e together with the nitrogen atom to which they are attached, R 7 forming a 4- to 6-membered heterocyclyl or a 5- to 7-membered heteroaryl, each optionally substituted with 1 to 3 groups selected from R fis selected from hydrogen, (C1-C4)alkyl, halo(C1-C4)alkyl, phenyl, (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclyl, wherein said phenyl, (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclyl are each selected from R 8 and optionally substituted with 1 to 3 groups selected from R a , R 6 , R 7 and R 8 are each independently selected from (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, (C3-C6)cycloalkyl, -NR'R'', -(C1-C4)alkylNR'R''-(C1-C4)alkylC(O)NR'R'', oxo, -(C1-C4)alkylOR', -C(O)R', -S(O)R' and -S(O)R' (or R' and R'' are each independently selected from hydrogen, (C1-C4) alkyl, and (C3-C6) cycloalkyl, where the remaining variables are as described above for Formula I or in any one of the second through fourth embodiments.

[0036] In a sixth embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof 2 is hydrogen, halo, (C1-C4) alkyl, (C3-C6) cycloalkyl, 5- to 7-membered heteroaryl, phenyl, 4- to 6-membered heterocyclyl, -NR a R b , -OR c , -C(O)NR d R e , -(C1-C4) alkylNR d R e and -(C1-C4) alkylC(O)NR d R ewherein each occurrence of 5- to 7-membered heteroaryl, phenyl, (C-C)cycloalkyl, and 4- to 6-membered heterocyclyl is selected from R 5 and optionally substituted with 1 to 3 groups selected from: 2 is hydrogen, halo, (C1-C4) alkyl, (C3-C6) cycloalkyl, 5- to 7-membered heteroaryl, phenyl, 4- to 6-membered heterocyclyl, -NR a R b , -OR c , -C(O)NR d R e , -(C1-C4) alkylNR d R e , -(C1-C4) alkylNR a C(O)R b and -(C1-C4) alkylC(O)NR d R e wherein each occurrence of 5- to 7-membered heteroaryl, phenyl, (C-C)cycloalkyl, and 4- to 6-membered heterocyclyl is selected from R 5 and optionally substituted with 1 to 3 groups selected from: 2 is hydrogen, halo, (C1-C4) alkyl, cyclopropyl, pyrazolyl, phenyl, pyrrolidinyl, piperazinyl, piperidinyl, -NR a R b , -OR c , -C(O)NR d R e , -(C1-C4) alkylNR d R e , -(C1-C4) alkylNR a C(O)R band -(C1-C4) alkylC(O)NR d R e wherein said cyclopropyl, pyrazolyl, phenyl, pyrrolidinyl, piperazinyl and piperidinyl are each selected from R 5 and optionally substituted with 1 to 3 groups selected from: 2 is hydrogen, halo, (C1-C4) alkyl, cyclopropyl, pyrazolyl, phenyl, pyrrolidinyl, piperazinyl, piperidinyl, -NR a R b , -OR c , -C(O)NR d R e , -(C1-C4) alkylNR d R e and -(C1-C4) alkylC(O)NR d R e wherein said cyclopropyl, pyrazolyl, phenyl, pyrrolidinyl, piperazinyl and piperidinyl are each selected from R 5 wherein the remaining variables are as described above for Formula I or in any one of the second through fifth embodiments.

[0037] In a seventh embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof a , R b and R c are each independently selected from hydrogen, (C1-C4) alkyl, -(C1-C4) alkyl[5- to 7-membered heteroaryl], phenyl, -(C1-C4) alkylphenyl, 4- to 6-membered heterocyclyl, and -(C1-C4) alkyl[4- to 6-membered heterocyclyl], wherein at each occurrence of 5- to 7-membered heteroaryl, 4- to 6-membered heterocyclyl, and phenyl, said groups are each independently selected from R 6and optionally substituted with 1 to 3 groups selected from: a , R b and R c are each independently selected from hydrogen, (C1-C4)alkyl, -(C1-C4)alkyl[5- to 7-membered heteroaryl], (C3-C6)cycloalkyl, phenyl, -(C1-C4)alkylphenyl, 4- to 6-membered heterocyclyl, and -(C1-C4)alkyl[4- to 6-membered heterocyclyl], wherein for each occurrence of 5- to 7-membered heteroaryl, 4- to 6-membered heterocyclyl, and phenyl, said groups are each independently selected from R 6 wherein the remaining variables are as described above for Formula I or in any one of the second through sixth embodiments.

[0038] In an eighth embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof 5 and R 6 are each independently selected from (C1-C4)alkyl and halo(C1-C4)alkyl, where the remaining variables are as described above for Formula I or in any one of the second through seventh embodiments. Alternatively, as part of the eighth embodiment, R in a compound of Formula I or a pharmaceutically acceptable salt thereof 5 and R 6 are each independently selected from halo, (C1-C4)alkyl, halo(C1-C4)alkyl, —(C1-C4)alkylOR′, and S(O)R′, where the remaining variables are as described above for Formula I or in any one of the second through seventh embodiments.

[0039] In a ninth embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof 2 is hydrogen, chloro, methyl, cyclopropyl, -OCH2CF3,

[0040] [ka]

[0041] wherein the remaining variables are as described above for Formula I or as described in any one of the second through eighth embodiments. Alternatively, as part of the ninth embodiment, R in a compound of Formula I or a pharmaceutically acceptable salt thereof is selected from: 2 is hydrogen, chloro, methyl, cyclopropyl, -OCH2CF3, -CH2NHC(O)CH3, -CH2NHSO2CH3, -CH2C(O)NHCH3, -CH2C(O)N(CH3)2, -CH2C(O)NHCH2CF3, -CH2C(O)NHCH2OCH3, -CH2C(O)NH2, -CH2C(O)N(CH3)(CH2CF3), -CH2C(O)NH(CH2)2SO2CH3, -C(O)N(CH3)2, -C(O)NHCH3,

[0042] [ka]

[0043] wherein the remaining variables are as described above for Formula I or as described in any one of the second through eighth embodiments. Alternatively, as part of the ninth embodiment, R in a compound of Formula I or a pharmaceutically acceptable salt thereof is selected from: 2 is selected from hydrogen, chloro, and —OCH2CF3, where the remaining variables are as described above for Formula I or in any one of the second through eighth embodiments. In another alternative, as part of the ninth embodiment, R in a compound of Formula I or a pharmaceutically acceptable salt thereof 2is hydrogen, where the remaining variables are as described above for Formula I or in any one of the second through eighth embodiments.

[0044] In a tenth embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof 3 is halo, (C1-C4) alkyl, halo(C1-C4) alkyl, (C3-C6) cycloalkyl, -(C1-C4) alkyl(C3-C6) cycloalkyl, 5- to 7-membered heteroaryl, -(C1-C4) alkyl [5- to 7-membered heteroaryl], phenyl, -(C1-C4) alkyl OR c1 , -(C1-C4) alkylphenyl, 4- to 6-membered heterocyclyl, -(C1-C4) alkyl[4- to 9-membered heterocyclyl], -NR a1 R b1 , -NHC(O)R c1 , -OR c1 , -(C1-C4) alkyl OR c1 , -C(O)R f1 , -C(O)NR d1 R e1 , -(C1-C4) alkylNR g1 R h1 , -(C1-C4) alkylC(O)R f1 , -(C1-C4) alkylC(O)NR d1 R e1 and -SR g1 wherein each occurrence of (C-C)cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclyl is selected from R 5a and optionally substituted with 1 to 3 groups selected from R a1 , R b1 and R c1are each independently selected from hydrogen, (C1-C4)alkyl, halo(C1-C4)alkyl, (C3-C6)cycloalkyl, -(C1-C4)alkyl(C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, -(C1-C4)alkyl[5- to 7-membered heteroaryl], phenyl, -(C1-C4)alkylphenyl, 4- to 6-membered heterocyclyl, -(C1-C4)alkyl[4- to 6-membered heterocyclyl], wherein at each occurrence of (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclyl, said groups each may be selected from R 6a and optionally substituted with 1 to 3 groups selected from R d1 , R e1 , R g1 and R h1 are each independently hydrogen, (C1-C4) alkyl, halo(C1-C4) alkyl, -(C1-C4) alkyl OR' 1 , -(C1-C4) alkylNR' 1 R'' 1 , -(C1-C4) alkylC(O)NR' 1 R'' 1 , -(C1-C4) alkylC(O)R' 1 , -(C1-C4) alkylC(O)OR' 1 , -(C1-C4) alkylS(O)R' 1 , -(C1-C4) alkylS(O)2R' 1 , (C1-C4) alkylphenyl, phenyl, (C3-C6) cycloalkyl, -(C1-C4) alkyl [5- to 7-membered heteroaryl], 5- to 7-membered heteroaryl, 4- to 6-membered heterocyclyl, -(C1-C4) alkyl [4- to 6-membered heterocyclyl], -C(O)R' 1 , -C(O)OR' 1 , -C(O)NR' 1 R'' 1 , -S(O)R' 1 and -S(O)R' 1 wherein each occurrence of (C-C)cycloalkyl, phenyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclyl is selected from R7a or R d1 and R e1 together with the nitrogen atom to which they are attached, R 7a forming a 4- to 6-membered heterocyclyl or a 5- to 7-membered heteroaryl, each optionally substituted with 1 to 3 groups selected from R f1 is selected from hydrogen, (C1-C4)alkyl, halo(C1-C4)alkyl, phenyl, (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclyl, wherein said phenyl, (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclyl are each selected from R 8a and optionally substituted with 1 to 3 groups selected from R 5a , R 6a , R 7a and R 8a are each independently halo, (C1-C4) alkyl, halo(C1-C4) alkyl, (C1-C4) alkoxy, halo(C1-C4) alkoxy, (C3-C6) cycloalkyl, cyano, or -NR' 1 R'' 1 , -(C1-C4) alkylNR' 1 R'' 1 , -(C1-C4) alkylC(O)NR' 1 R'' 1 , oxo, -(C1-C4) alkyl OR' 1 , -C(O)R' 1 , -S(O)R' 1 and -S(O)R' 1 is selected from R' 1 and R'' 1are each independently selected from hydrogen, (C1-C4) alkyl, (C3-C6) cycloalkyl, and 4- to 7-membered heterocyclyl, where the remaining variables are as described above for Formula I or in any one of the second through ninth embodiments. Alternatively, as part of the tenth embodiment, R in a compound of Formula I or a pharmaceutically acceptable salt thereof 3 is halo, (C1-C4) alkyl, halo(C1-C4) alkyl, (C3-C6) cycloalkyl, -(C1-C4) alkyl(C3-C6) cycloalkyl, 5- to 7-membered heteroaryl, -(C1-C4) alkyl [5- to 7-membered heteroaryl], phenyl, -(C1-C4) alkyl OR c1 , -(C1-C4) alkylphenyl, 4- to 10-membered heterocyclyl, -(C1-C4) alkyl[4- to 10-membered heterocyclyl], -NR a1 R b1 , -NHC(O)R c1 , -OR c1 , -(C1-C4) alkyl OR c1 , -C(O)R f1 , -C(O)NR d1 R e1 , -(C1-C4) alkylNR g1 R h1 , -(C1-C4) alkylC(O)R f1 , -(C1-C4) alkylC(O)NR d1 R e1 and -SR g1 wherein each occurrence of (C-C)cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 10-membered heterocyclyl is selected from R 5a and optionally substituted with 1 to 3 groups selected from R a1 , R b1 and R c1are each independently selected from hydrogen, (C1-C4)alkyl, halo(C1-C4)alkyl, (C3-C6)cycloalkyl, -(C1-C4)alkyl(C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, -(C1-C4)alkyl[5- to 7-membered heteroaryl], phenyl, -(C1-C4)alkylphenyl, 4- to 6-membered heterocyclyl, -(C1-C4)alkyl[4- to 6-membered heterocyclyl], wherein at each occurrence of (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclyl, said groups each may be selected from R 6a and optionally substituted with 1 to 3 groups selected from R d1 , R e1 , R g1 and R h1 are each independently hydrogen, (C1-C4) alkyl, halo(C1-C4) alkyl, -(C1-C4) alkyl OR' 1 , -(C1-C4) alkylNR' 1 R'' 1 , -(C1-C4) alkylC(O)NR' 1 R'' 1 , -(C1-C4) alkylC(O)R' 1 , -(C1-C4) alkylC(O)OR' 1 , -(C1-C4) alkylS(O)R' 1 , -(C1-C4) alkylS(O)2R' 1 , (C1-C4) alkylphenyl, phenyl, (C3-C6) cycloalkyl, -(C1-C4) alkyl [5- to 7-membered heteroaryl], 5- to 7-membered heteroaryl, 4- to 8-membered heterocyclyl, -(C1-C4) alkyl [4- to 8-membered heterocyclyl], -C(O)R' 1 , -C(O)OR' 1 , -C(O)NR' 1 R'' 1 , -S(O)R' 1 and -S(O)R' 1 wherein each occurrence of (C-C)cycloalkyl, phenyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclyl is selected from R7a or R d1 and R e1 together with the nitrogen atom to which they are attached, R 7a forming a 4- to 8-membered heterocyclyl or a 5- to 7-membered heteroaryl, each optionally substituted with 1 to 3 groups selected from R f1 is selected from hydrogen, (C1-C4)alkyl, halo(C1-C4)alkyl, phenyl, (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclyl, wherein said phenyl, (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclyl are each selected from R 8a and optionally substituted with 1 to 3 groups selected from R 5a , R 6a , R 7a and R 8a are each independently halo, (C1-C4) alkyl, halo(C1-C4) alkyl, (C1-C4) alkoxy, halo(C1-C4) alkoxy, (C3-C6) cycloalkyl, 5- to 7-membered heteroaryl, cyano, or -NR' 1 R'' 1 , -(C1-C4) alkylNR' 1 R'' 1 , -(C1-C4) alkylC(O)OR' 1 , -(C1-C4) alkylC(O)NR' 1 R'' 1 , oxo, -(C1-C4) alkyl OR' 1 , -C(O)NR' 1 R'' 1 , -OR' 1 , C(O)OR' 1 , -C(O)R' 1 , -S(O)R' 1 , -S(O)2R' 1 , R' 1 and 4- to 6-membered heterocyclyl optionally substituted with 1 to 3 groups selected from R' 1and R'' 1 are each independently selected from hydrogen, (C1-C4) alkyl, (C3-C6) cycloalkyl, and 4- to 7-membered heterocyclyl, where the remaining variables are as described above for Formula I or in any one of the second through ninth embodiments.

[0045] In an eleventh embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof 3 is -(C1-C4) alkyl [4- to 9-membered heterocyclyl], -(C1-C4) alkyl [5- to 7-membered heteroaryl], 4- to 6-membered heterocyclyl, -(C1-C4) alkyl OR c1 , -NR a1 R b1 , -NHC(O)R c1 , -C(O)R f1 , -C(O)NR d1 R e1 and -(C1-C4) alkylNR g1 R h1 wherein the (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, 4- to 6-membered heterocyclyl, and 4- to 9-membered heterocyclyl are each selected from R 5a and R in the compound of Formula I or a pharmaceutically acceptable salt thereof is optionally substituted with one to three groups selected from: 3 is -(C1-C4) alkyl [4- to 10-membered heterocyclyl], -(C1-C4) alkyl [5- to 7-membered heteroaryl], 4- to 10-membered heterocyclyl, -(C1-C4) alkyl OR c1 , -NR a1 R b1 , -NHC(O)R c1 , -C(O)R f1 , -C(O)NR d1 R e1 and -(C1-C4) alkylNR g1 R h1wherein the (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, 4- to 10-membered heterocyclyl, and 4- to 9-membered heterocyclyl are each selected from R 5a and R in the compound of Formula I or a pharmaceutically acceptable salt thereof is optionally substituted with one to three groups selected from: 3 is -(C1-C4) alkyl OR c1 , 4- to 10-membered heterocyclyl, -(C1-C4)alkyl[4- to 10-membered heterocyclyl], -NR a1 R b1 , -NHC(O)R c1 , -C(O)R f1 , -C(O)NR d1 R e1 , -(C1-C4) alkylNR g1 R h1 wherein each 4- to 10-membered heterocyclyl is selected from R 5a and R in the compound of Formula I or a pharmaceutically acceptable salt thereof is optionally substituted with one to three groups selected from: 3 is -(C1-C4) alkyl OR c1 , 4- to 6-membered heterocyclyl, -(C1-C4)alkyl[4- to 9-membered heterocyclyl], -NR a1 R b1 , -NHC(O)R c1 , -C(O)R f1 , -C(O)NR d1 R e1 , -(C1-C4) alkylNR g1 R h1 wherein the 4- to 6-membered heterocyclyl and the 4- to 9-membered heterocyclyl are each selected from R 5aand optionally substituted with 1 to 3 groups selected from: 3 is -(C1-C4) alkyl OR c1 , -NR a1 R b1 , -NHC(O)R c1 , -C(O)R f1 , -C(O)NR d1 R e1 , -(C1-C4) alkylNR g1 R h1 , -(C1-C4)alkyl[piperazinyl], -(C1-C4)alkyl[piperidinyl], -(C1-C4)alkyl[morpholinyl], -(C1-C4)alkyl[pyrrolidinyl], -(C1-C4)alkyl[diazepanyl], -(C1-C4)alkyl[azetidinyl], piperazinyl, and tetrahydropyridinyl, wherein said piperidinyl, morpholinyl, pyrrolidinyl, diazepanyl, tetrahydropyridinyl, azetidinyl, and each occurrence of piperazinyl is selected from R 5a wherein the remaining variables are as described above for Formula I or in any one of the second through tenth embodiments.

[0046] In a twelfth embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof 5a is (C1-C4) alkyl, halo(C1-C4) alkyl, (C3-C6) cycloalkyl, -NR' 1 R'' 1 , -(C1-C4) alkylNR' 1 R'' 1 , -(C1-C4) alkyl OR' 1 and -C(O)R' 1wherein the remaining variables are as described above for Formula I or as described in any one of the second through eleventh embodiments. Alternatively, as part of the twelfth embodiment, R in a compound of Formula I or a pharmaceutically acceptable salt thereof is selected from: 5a is selected from (C1-C4)alkyl, halo(C1-C4)alkyl, cyclopropyl, —N[(C1-C4)alkyl]2, —(C1-C4)alkylN[(C1-C4)alkyl]2, —(C1-C4)alkylOH, —(C1-C4)alkylO(C1-C4)alkyl, and —C(O)(C1-C4)alkyl, where the remaining variables are as described above for Formula I or as described in any one of the second through eleventh embodiments.

[0047] In a thirteenth embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof c1 is selected from 5- to 7-membered heteroaryl and 4- to 6-membered heterocyclyl, where the 5- to 7-membered heteroaryl and the 4- to 6-membered heterocyclyl are, respectively, R 6a and optionally substituted with 1 to 3 groups selected from: c1 is R 6a wherein the remaining variables are as described above for Formula I or in any one of the second through twelfth embodiments.

[0048] In a fourteenth embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof 6a is (C1-C4) alkyl, where the remaining variables are as described above for Formula I or in any one of the second through thirteenth embodiments.

[0049] In a fifteenth embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof a1 and R b1 are each independently selected from hydrogen, —(C1-C4)alkyl[5- to 7-membered heteroaryl], and 4- to 6-membered heterocyclyl, wherein at each occurrence of 5- to 7-membered heteroaryl and 4- to 6-membered heterocyclyl, said groups are each independently selected from R 6a and optionally substituted with 1 to 3 groups selected from: a1 and R b1 are each independently selected from hydrogen and -(C1-C4) alkyl [5- to 7-membered heteroaryl], wherein said 5- to 7-membered heteroaryl is R 6a and optionally substituted with 1 to 3 groups selected from: a1 is hydrogen and R b1 is -(C1-C4) alkyl[pyridinyl], where the pyridinyl is R 6a wherein the remaining variables are as described above for Formula I or in any one of the second through fourteenth embodiments.

[0050] In a sixteenth embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof f1 is R 8aand R is a 4- to 6-membered heterocyclyl optionally substituted with 1 to 3 groups selected from: f1 is R 8a wherein the remaining variables are as described above for Formula I or in any one of the second through fifteenth embodiments.

[0051] In a seventeenth embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof 8a is (C1-C4) alkyl, where the remaining variables are as described above for Formula I or in any one of the second through sixteenth embodiments.

[0052] In an eighteenth embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof d1 and R e1 are each independently hydrogen, -(C1-C4) alkylNR' 1 R'' 1 , (C1-C4) alkyl [5- to 7-membered heteroaryl] and 4- to 6-membered heterocyclyl, wherein said 5- to 7-membered heteroaryl and 4- to 6-membered heterocyclyl are, respectively, selected from R 7a and optionally substituted with 1 to 3 groups selected from: d1 and R e1 are each independently hydrogen, -(C1-C4) alkylNR' 1 R'' 1, (C1-C4) alkyl [5- to 7-membered heteroaryl] and 4- to 8-membered heterocyclyl, wherein said 5- to 7-membered heteroaryl and 4- to 8-membered heterocyclyl are each selected from R 7a and optionally substituted with 1 to 3 groups selected from: d1 and R e1 are each independently selected from hydrogen, —(C1-C4)alkylN[(C1-C4)alkyl]2, (C1-C4)alkyl[pyridinyl], and piperidinyl, wherein said pyridinyl and piperidinyl are each independently selected from R 7a wherein the remaining variables are as described above for Formula I or in any one of the second through seventeenth embodiments.

[0053] In a nineteenth embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof g1 and R h1 are each independently hydrogen, (C1-C4) alkyl, halo(C1-C4) alkyl, -(C1-C4) alkyl OR' 1 , -(C1-C4) alkylNR' 1 R'' 1 , -(C1-C4) alkylC(O)NR' 1 R'' 1 , -(C1-C4) alkylC(O)R' 1 , (C1-C4) alkylphenyl, -(C1-C4) alkyl[5- to 7-membered heteroaryl], and 4- to 6-membered heterocyclyl, wherein said phenyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclyl are each selected from R 7aand optionally substituted with 1 to 3 groups selected from: g1 and R h1 are each independently hydrogen, (C1-C4) alkyl, halo(C1-C4) alkyl, -(C1-C4) alkyl OR' 1 , -(C1-C4) alkylNR' 1 R'' 1 , -(C1-C4) alkylC(O)NR' 1 R'' 1 , -(C1-C4) alkylC(O)R' 1 , (C1-C4) alkylphenyl, -(C1-C4) alkyl[5- to 7-membered heteroaryl], and 4- to 8-membered heterocyclyl, wherein said phenyl, 5- to 7-membered heteroaryl, and 4- to 8-membered heterocyclyl are each selected from R 7a and optionally substituted with 1 to 3 groups selected from: g1 and R h1 are each independently selected from hydrogen, (C1-C4)alkyl, halo(C1-C4)alkyl, -(C1-C4)alkyl[pyridinyl], -(C1-C4)alkylO(C1-C4)alkyl, (C1-C4)alkylphenyl, -(C1-C4)alkylN[(C1-C4)alkyl]2, pyridinyl, piperidinyl, pyrrolidinyl, -(C1-C4)alkylC(O)N[(C1-C4)alkyl]2, halo(C1-C4)alkyl, -(C1-C4)alkylC(O)(morpholinyl), wherein said phenyl, pyridinyl, piperidinyl, pyrrolidinyl and morpholinyl are each independently selected from R 7aand optionally substituted with 1 to 3 groups selected from: g1 and R h1 are each independently selected from hydrogen, (C1-C4)alkyl, halo(C1-C4)alkyl, -(C1-C4)alkyl[pyridinyl], -(C1-C4)alkylO(C1-C4)alkyl, (C1-C4)alkylphenyl, -(C1-C4)alkylN[(C1-C4)alkyl]2, piperidinyl, pyrrolidinyl, -(C1-C4)alkylC(O)N[(C1-C4)alkyl]2, halo(C1-C4)alkyl, -(C1-C4)alkylC(O)(morpholinyl), wherein said phenyl, pyridinyl, piperidinyl, pyrrolidinyl and morpholinyl are each independently selected from R 7a and optionally substituted with 1 to 3 groups selected from: 7a is selected from (C1-C4)alkyl, halo(C1-C4)alkyl, halo, (C1-C4)alkoxy, and cyano, where the remaining variables are as described above for Formula I or in any one of the second through eighteenth embodiments.

[0054] In a twentieth embodiment, R in a compound of formula I or a pharmaceutically acceptable salt thereof 3 -CH2NHCH3, -CH2N(CH3)2,

[0055] [ka]

[0056] , -CH2N(CH3)(CH2)2OCH3, -CH2N(CH3)(CH2)2N(CH3)2, -CH2N(CH3)(CH2)3N(CH3)2, -CH2N(CH3)CH2C(O)N(CH3)2, -C(O)NHCH2N(CH3)2,

[0057] [ka]

[0058] wherein the remaining variables are as described above for Formula I or as described in any one of the second through nineteenth embodiments. Alternatively, as part of the twentieth embodiment, R in a compound of Formula I or a pharmaceutically acceptable salt thereof is selected from: 3 is -CH2NHCH3, -CH2N(CH3)2, -(CH2)2N(CH3)2, -CH2N(CH3)(CH2)2OCH3, -CH2N(CH3)(CH2)2N(CH3)2, -CH2NH(CH2CH3), -C H2N(CH3)(CH2)3N(CH3)2, -CH2NH(CH2)2N(CH2CH3)2, CH2NHC(CH3)3, -CH2N(CH3)CH2C(O)N(CH3)2, -C(O)NHCH2N(CH3)2,

[0059] [ka]

[0060] [ka]

[0061] [ka]

[0062] [ka]

[0063] [ka]

[0064] wherein the remaining variables are as described above for Formula I or in any one of the second through nineteenth embodiments. Compounds having Formula I are further disclosed in the examples and are included in this disclosure, including pharmaceutically acceptable salts thereof, as well as neutral forms.

[0065] 4. Use, Formulation and Administration The compounds and compositions described herein are generally useful for modifying the activity of TP53. In some embodiments, the compounds, pharmaceutically acceptable salts, and pharmaceutical compositions described herein are covalent modifiers of Y220C. In some embodiments, the compounds, pharmaceutically acceptable salts, and pharmaceutical compositions described herein stabilize TP53. In some embodiments, the compounds, pharmaceutically acceptable salts, and pharmaceutical compositions described herein restore wild-type tumor suppressor protein p53 (WT TP53) function.

[0066] In some embodiments, the compounds and pharmaceutical compositions described herein are useful for treating conditions associated with TP53 function. In some embodiments, the compounds and pharmaceutical compositions described herein are useful for treating conditions responsive to activation of TP53 function. In some embodiments, the compounds and pharmaceutical compositions described herein are useful for treating conditions responsive to restoration of TP53 function where protein function is inactivated due to a mutation, such as the Y220C mutation.

[0067] In some embodiments, the compounds and pharmaceutical compositions described herein are useful for treating cancer. In some embodiments, the compounds and pharmaceutical compositions described herein are useful for treating cancers that express TP53 mutants (e.g., cancers harboring the Y220C mutation).

[0068]

[0055] Particular cancers treatable by the present compounds, salts and compositions include, but are not limited to, solid tumors, hemolytic malignancies, ovarian cancer, esophageal cancer, colon cancer, head and neck cancer, laryngeal cancer, lung cancer, leukemia (e.g., acute myeloid leukemia (AML)), sarcoma, testicular cancer, melanoma, cervical cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, endometrial cancer, esophageal cancer, gastric cancer, prostate cancer, bladder cancer, myelodysplastic syndrome (MDS), sarcoma and melanoma.

[0069]

[0056] Also provided is the use of a compound disclosed herein or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the disclosed compound or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating the described conditions. Further provided is a compound disclosed herein or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the disclosed compound or a pharmaceutically acceptable salt thereof, for use in treating the described conditions.

[0070] In certain aspects, the pharmaceutical compositions described herein are formulated for administration to a patient in need of such compositions. The pharmaceutical compositions described herein may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or via an implanted reservoir. As used herein, the term "parenteral" includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. In some embodiments, the compositions are administered orally, intraperitoneally, or intravenously. Sterile injectable forms of the pharmaceutical compositions described herein may be aqueous or oleaginous suspensions. These suspensions may be formulated according to techniques known to those skilled in the art using suitable dispersing or wetting agents and suspending agents.

[0071] In some embodiments, the pharmaceutical composition is administered orally. The specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the particular compound utilized, age, body weight, general health, sex, diet, time of administration, excretion rate, drug combination, and the judgment of the treating physician, the severity of the particular disease being treated. The amount of a compound described herein in a composition will also depend on the particular compound in the pharmaceutical composition. Example chemical synthesis

[0060] The following representative examples are intended to help illustrate the present disclosure and are not intended to, nor should they be construed to, limit the scope of the invention.

[0072]

[0061]

[0073] [ka]

[0074] 4-Chloropyrimidoindoles such as S10 can be prepared via a nine-step process beginning with the appropriate fluorocyanobenzoate or fluorocyanonicotinate ester, represented by S1. Nucleophilic aromatic substitution of S1 with a glycinate ester provides intermediate S2, which can be converted to the 3-amino-indole derivative S3. Concentration of S3 with dimethylformamide dimethylacetyl followed by treatment with ammonia produces the hydroxypyrimidoindole intermediate S5. S5 can be converted to the chloride intermediate S6 with phosphorus oxychloride. Alkylation of S6 with an appropriate electrophile provides S7. Reduction of S7 provides the benzyl alcohol intermediate S8, which can be converted to the benzyl chloride intermediate S9 with thionyl chloride. Alkylation of an appropriate amine with S9 provides the target compound, such as S10.

[0075]

[0063]

[0076] [ka]

[0077] 4-Chloropyrimidindoles such as S14 can be prepared from intermediates such as S11 via a three-step process. Saponification of S11 provides the acid intermediate S12, which can be converted to the amide target compounds such as S14 via conversion to the acid chloride intermediate S13 followed by reaction with an amine.

[0078]

[0065]

[0079] [ka]

[0080] 4-Chloropyrimidindoles such as S17 can be prepared from intermediates such as S15 via a two-step process. Oxidation of S15 leads to intermediate S16, which can be converted to target compounds such as S17 via reductive amination.

[0081]

[0067]

[0082] [ka]

[0083] 4-Chloropyrimidindoles such as S22 can be prepared from intermediates such as S18 via a four-step process. Treatment of S18 with trimethylhydroxytin affords acid intermediates such as S19, which can be converted to amine intermediates S21 via a two-step process involving Curtius rearrangement and acid-mediated removal of the Boc protecting group. Derivatization of S21 by reductive amination or amide coupling affords the target compounds represented by S22.

[0084]

[0069]

[0085] [ka]

[0086] Chloropyrimidindoles such as S32 can be prepared via a nine-step process starting with an appropriate cyano-fluoro-iodo-benzene such as S23. Nucleophilic aromatic substitution of S23 with ethyl glycinate provides S24, which can be converted to the 3-amino-indole intermediate S25 by treatment with BocO and DMAP. In a manner similar to Scheme 1, S25 can be converted to intermediate S29 via a four-step process. Treatment of S29 with sodium methoxide produces a methoxypyrimidoindole intermediate such as S30. Palladium-catalyzed amination provides the iodide intermediate S31, which can be converted to the target compound such as S32 by treatment with phosphorus oxychloride.

[0087]

[0071]

[0088] [ka]

[0089] Dichloropyrimidoindoles such as S40 can be prepared via a seven-step process starting with the appropriate bromo-cyano-fluoro-benzene, such as S33. Treatment of S33 with aminoacetamide followed by base-mediated cyclization affords 3-amino-indole intermediates, such as S35. Treatment of S35 with triphosgene affords dihydroxypyrimidoindole intermediates, such as S36, which can be converted to the desired target compounds, such as S40, via a four-step sequence similar to that described in Scheme 1.

[0090]

[0073]

[0091] [ka]

[0092] Chloropyrimidindoles such as S46 can be prepared via a five-step sequence starting with intermediate S41. Alkylation of S41 with an electrophile provides intermediate S42. Selective displacement of the chloride with sodium methoxide provides intermediate S43. S43 can be converted to S45 via Suzuki coupling with a boronic acid or ester to provide intermediate S45, which can then be converted to the target compound S46 by treatment with phenylphosphonic acid dichloride.

[0093]

[0075]

[0094] [ka]

[0095] Chloropyrimidoindoles such as S55 can be prepared via an eight-step sequence starting with intermediate S47. Diamide S48 can be prepared by coupling Boc-glycine with 3-aminoindole S47. Base-mediated cyclization of S48 provides hydroxypyrimidinol intermediate S49, which can be converted to chloropyrimidine S50 with POCl. After protecting the free amine in S50 with a Boc group, the indole can be alkylated with an electrophile to provide intermediate S52. Suzuki coupling of S52 with potassium ((dimethylamino)methyl)trifluoroborate followed by treatment with POCl provides intermediate S54, which can be converted to target compounds such as S55 by amide coupling with a carboxylic acid or reaction with an acid chloride.

[0096]

[0077]

[0097] [ka]

[0098] Chloropyrimidoindoles such as S64 can be prepared via an eight-step sequence starting with intermediate S56. Reaction of 3-amino-2-carboxamidoindole intermediate S56 with diethyl malonate provides intermediate S57, which can be converted to chloropyrimidine intermediate S58 using POCl. Alkylation of S58 with an electrophile provides intermediate S59. Displacement of the chloride with sodium methoxide provides S60, which can be converted to 7-diaminomethylpyrimidoindole intermediate S61 via Suzuki coupling with potassium ((dimethylamino)methyl)trifluoroborate. Treatment of S61 with POCl provides intermediate S62. Hydrolysis of ester S62 with trimethyltin hydroxide provides carboxylic acid intermediate S63, which can be coupled with an amine nucleophile to provide target compounds such as S64.

[0099]

[0079]

[0100] [ka]

[0101] Chloropyrimidindoles such as S73 can be prepared via a nine-step sequence starting from intermediate S65. Reaction of 3-amino-2-carboxamidoindole intermediate S65 with diethyl oxalate provides intermediate S66, which can be converted to methoxypyrimidine intermediate S67 by sequential treatment with POCl and sodium methoxide. Alkylation of S67 with electrophiles provides intermediate S68, which can be converted to 7-diaminomethylpyrimidoindole intermediate S69 via Suzuki coupling with potassium ((dimethylamino)methyl)trifluoroborate. Sequential treatment of S69 with oxalyl chloride followed by methanol provides methyl ester intermediate S70, which can be selectively demethylated with trimethylsilyl chloride and sodium iodide to provide hydroxypyrimidoindole intermediate S71. Sequential treatment of S71 with POCl3 and trimethylhydroxytin affords the carboxylic acid intermediate S73, which can be converted to target compounds such as S74 via amide coupling.

[0102]

[0081]

[0103] [ka]

[0104] Chloropyrimidindoles such as S77 can be prepared via a two-step sequence starting with intermediate S75. Alkylation of the amine with intermediate S75 leads to intermediate S76, which can be converted to the target compound such as S77 via treatment with hydrochloric acid.

[0105]

[0083]

[0106] [ka]

[0107] Chloropyrimidindoles such as S80 can be prepared via a two-step sequence starting with intermediate S78. Reductive amination with intermediate S75 affords intermediate S79, which can be converted to target compounds such as S80 via treatment with hydrochloric acid.

[0108]

[0085]

[0109] [ka]

[0110] Chloropyrimidindoles such as S90 can be prepared via a nine-step sequence starting with intermediate S81. N Ar reaction followed by deprotection with TFA affords the aniline intermediate S83. Reductive amination of S83 with ethyl glyoxalate affords S84. Treatment of S84 with Boc-anhydride and 4-dimethylaminopyridine induces a cyclization reaction, which produces the indole intermediate S85. Sequential treatment of S85 with DMF-DMA and ammonia affords the hydropyrimidinol intermediate S87. Treatment of S87 with POCl3 affords the chloropyrimidoindole intermediate S88. Alkylation of S88 with electrophiles followed by Suzuki coupling with potassium ((dimethylamino)methyl)trifluoroborate affords the target compounds such as S90.

[0111]

[0087]

[0112] [ka]

[0113] Chloropyrimidindoles such as S94 can be prepared via one of two different two-step sequences starting with intermediates such as S91. When X = OM, palladium-catalyzed cross-coupling of S91 with a boronic acid or boronic ester provides intermediate S92, which can be converted to target compounds such as S94 by treatment with POCl3. When X = Cl, palladium-catalyzed borylation of S91 followed by Suzuki coupling with a boronic acid or boronic ester provides target compounds such as S94.

[0114]

[0089] Abbreviation: ACN = acetonitrile AcOH = acetic acid Boc2O = di-tert-butyl dicarbonate DCM = dichloromethane DCE = 1,2-dichloroethane DIEA = N,N-diisopropylethylamine DMAP = 4-dimethylaminopyridine DMF = dimethylformamide DMF-DMA = N,N-dimethylformamide dimethyl acetal DPPA = diphenylphosphoryl azide EtOAc = ethyl acetate EtOH = ethanol IPA = Isopropanol MeOH = methanol TEA = triethylamine TFA = trifluoroacetic acid THF = tetrahydrofuran TMSCl = trimethylsilyl chloride TMSBr = trimethylsilyl bromide

[0115] Example 1 1-(4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)-N,N-dimethylmethanamine

[0116] [ka]

[0117] Scheme 1, Step 1. Methyl 3-cyano-4-[(2-ethoxy-2-oxo-ethyl)amino]benzoate: To a solution of ethyl 2-aminoacetate hydrochloride (8.6 g, 62.2 mmol, 1.0 equiv.) in ACN (110 mL) was added K2CO3 (20.4 g, 148.1 mmol, 2.5 equiv.) and methyl 3-cyano-4-fluorobenzoate (10.6 g, 59.1 mmol, 1.0 equiv.). The mixture was stirred at 90 °C for 4 h. The reaction mixture was poured into HO (80 mL), and the mixture was extracted with ethyl acetate (2 × 100 mL). The organic extract was washed with brine (100 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (3:1 petroleum ether:ethyl acetate) to afford the title compound (3.7 g, 14.1 mmol, 24% yield) as a white solid.

[0118]

[0092] 1 H NMR (400 MHz, chloroform-d) δ 8.09 - 8.06 (m, 1H), 7.99 (dd, J = 2.0, 8.8 Hz, 1H), 6.49 (d, J = 8.8 Hz, 1H), 5.53 (t, J = 4.4 Hz, 1H), 4.22 (q, J = 7.2 Hz, 2H), 3.97 (d, J = 5.2 Hz, 2H), 3.81 (s, 3H), 1.25 (t, J = 7.2 Hz, 3H).

[0093] LCMS [M-1] = 261.1.

[0119] Scheme 1, Step 2. 1-(tert-butyl) 2-ethyl 5-methyl 3-amino-1H-indole-1,2,5-tricarboxylate: To a solution of methyl 3-cyano-4-[(2-ethoxy-2-oxo-ethyl)amino]benzoate (2.7 g, 10.3 mmol, 1.0 equiv) in DCM (1.0 mL) was added TEA (1.06 g, 10.4 mmol, 1.4 mL, 1.0 equiv), BocO (2.7 g, 12.4 mmol, 1.2 equiv), and DMAP (125.7 mg, 1.0 mmol, 0.1 equiv). The reaction mixture was stirred at 20° C. for 2 hours. The reaction mixture was poured into water (20 mL) and extracted with EtOAc (2×20 mL). The organic extract was washed with brine (20 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (4.5:1 petroleum ether: EtOAc) to afford the title compound (4.4 g, crude) as a white solid.

[0120]

[0096] 1 HNMR (400 MHz, chloroform-d) δ 8.37–8.30 (m, 1H), 8.23 ​​(dd, J = 2.0, 8.4 Hz, 1H), 7.61 (s, 1H), 4.36 (s, 2H), 4.24 (q, J = 7.2 Hz, 2H), 3.96 (s, 3H), 1.54–1.40 (m, 9H), 1.34–1.27 (m, 3H).

[0097] LCMS [M-55] = 307.2.

[0121] Scheme 1, Step 3. 1-(tert-butyl) 2-ethyl 5-methyl 3-(((dimethylamino)methylene)amino)-1H-indole-1,2,5-tricarboxylate: To a solution of 1-(tert-butyl) 2-ethyl 5-methyl 3-amino-1H-indole-1,2,5-tricarboxylate (3.4 g, 9.3 mmol, 1.0 equiv.) in DMF (34 mL) was added DMF-DMA (1.6 mL, 12.2 mmol, 1.3 equiv.), and the mixture was stirred at 100° C. for 4 h. The reaction mixture was poured into HO (50 mL) and extracted with EtOAc (3×30 mL). The organic extract was washed with brine (20 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (4.5:1 petroleum ether:EtOAc) to afford the title compound (4 g, crude) as a white solid.

[0122]

[0100] 1 H NMR (400 MHz, chloroform-d) δ 8.39 (s, 1H), 8.12–8.03 (m, 2H), 7.93 (s, 1H), 4.30 (q, J = 6.8 Hz, 2H), 3.94 (s, 3H), 3.11 (d, J = 9.4 Hz, 6H), 1.63 (s, 9H), 1.33 (t, J = 7.2Hz, 3H).

[0101] LCMS [M+1] = 418.4.

[0123] Scheme 1, Step 4. Methyl 4-oxo-3,5-dihydropyrimido[5,4-b]indole-8-carboxylate: To a solution of 1-(tert-butyl)-2-ethyl-5-methyl-3-(((dimethylamino)methylene)amino)-1H-indole-1,2,5-tricarboxylate (3.6 g, 8.6 mmol, 1.0 equiv) in EtOH (40 mL) was added NH3·H2O (40 mL). The mixture was stirred at 70 °C for 16 h. The reaction mixture was filtered and the filter cake was concentrated in vacuo to give the title compound (1.2 g, crude) as a white solid. This material was used in the next step without further purification.

[0124]

[0104] 1 H NMR (400 MHz, DMSO-d6) δ 12.5 (s, 2H), 8.62 (s, 1H), 8.11 - 7.99 (m, 2H), 7.60 (d, J = 8.8 Hz, 1H), 3.88 (s, 3H). LCMS [M+1] = 244.2.

[0125] Scheme 1, Step 5. Methyl 4-chloro-5H-pyrimido[5,4-b]indole-8-carboxylate: A solution of methyl 4-hydroxy-5H-pyrimido[5,4-b]indole-8-carboxylate (900.0 mg, 3.7 mmol, 1.0 equiv) in POCl3 (18.0 mL) was stirred at 110°C for 20 h. The reaction mixture was cooled to RT and concentrated under reduced pressure. The crude product was triturated with ACN at 20°C for 10 min to give a residue. The residue was triturated with HO at 20°C for 2 min to give the title compound (600 mg, 62% yield) as a yellow solid.

[0126]

[0108] 1 H NMR (400 MHz, chloroform-d) δ 12.8 (s, 1H), 8.93 (s, 1H), 8.53 (s, 1H), 8.27 (d, J = 8.8 Hz 1H), 7.85 (d, J = 8.8 Hz, 1H), 3.92 (s, 3H).

[0109] LCMS[M+1, M+3]=262.2, 264.2.

[0127] Scheme 1, Step 6. Methyl 4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole-8-carboxylate: To a solution of methyl 4-chloro-5H-pyrimido[5,4-b]indole-8-carboxylate (500.0 mg, 1.9 mmol, 1.0 equiv) in dry DMF (5.0 mL) in a dry round-bottom flask was added NaH (115 mg, 2.8 mmol, 60% purity, 1.5 equiv) at 0° C. The reaction mixture was stirred under nitrogen at 20° C. for 0.5 h, and then 2,2,2-trifluoroethyl trifluoromethanesulfonate (887 mg, 3.8 mmol, 2.0 equiv) was added to the reaction mixture. After stirring at 20° C. for 16 h, the reaction mixture was quenched by the addition of HO (10 mL) at 0° C. The mixture was extracted with EtOAc (20 mL), and the organic extract was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (9:1 petroleum ether: EtOAc) to afford the title compound (330 mg, 50% yield) as a white solid.

[0128]

[0112] 1 H NMR (400 MHz, chloroform-d) δ 9.30 (s, 1H), 8.99 (s, 1H), 8.46 (dd, J = 1.6, 8.8 Hz, 1H), 7.63 (d, J = 8.8 Hz, 1H), 5.39 (q, J = 8.0 Hz, 2H), 4.01 (s, 3H). LCMS [M+1] = 344.2.

[0129] Scheme 1, Step 7. (4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)methanol: A 100 mL three-neck flask equipped with a stir bar was charged with methyl 4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole-8-carboxylate (5.0 g, 14.5 mmol, 1.0 equiv) and toluene (60 mL). The suspension was degassed under vacuum and purged with N2 three times. The solution was cooled to -60 °C, and then a solution of DIBAL-H (1 M, 29.1 mL, 2.0 equiv) was added dropwise over 15 min. The resulting solution was stirred under N2 at -60 °C for 1 h. The reaction mixture was quenched by the addition of EtOAc (500 mL) and then diluted with a saturated aqueous solution of potassium sodium tartrate (500 mL) and extracted with EtOAc (2 x 100 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (100:1 to 0:1 petroleum ether:EtOAc) to afford the title compound (3.0 g, 65% yield) as an off-white solid.

[0130]

[0116] 1 H NMR (400 MHz, chloroform-d) δ 8.78 (s, 1H), 8.27 (s, 1H), 7.71 (dd, J = 1.2, 8.4 Hz, 1H), 7.48 (d, J = 8.4 Hz, 1H), 5.26 (q, J = 8.4 Hz, 2H), 4.75 (s, 2H). LCMS: [M+1]=316.1.

[0131] Scheme 1, Step 8. 4-Chloro-8-(chloromethyl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole: To a solution of (4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)methanol (2.5 g, 7.9 mmol, 1.0 equiv) in toluene (40 mL) was added SOCl (3.8 g, 31.6 mmol, 4.0 equiv) under a nitrogen atmosphere at 0 °C. The reaction mixture was stirred at RT for 1 h. Most of the SOCl was removed under reduced pressure, and then the mixture was quenched by the dropwise addition of HO (20 mL). The mixture was extracted with EtOAc (3 × 20 mL). The combined organic extracts were dried over NaSO, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (100:1 to 0:1 petroleum ether:EtOAc) to afford the title compound (2.0 g, 76% yield) as an off-white solid.

[0132]

[0120] 1 H NMR (400 MHz, chloroform-d) δ 8.94 (s, 1H), 8.42 (d, J =1.2 Hz, 1H), 7.82 (dd, J = 1.6, 8.8 Hz, 1H), 7.58 (d, J = 8.8 Hz, 1H), 5.34 (q, J = 8.0 Hz, 2H), 4.81 (s, 2H). LCMS: [M+1]=334.1.

[0133] Scheme 1, Step 9. 1-(4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)-N,N-dimethylmethanamine: To a solution of 4-chloro-8-(chloromethyl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole (500.0 mg, 1.5 mmol, 1.0 equiv) in ACN (10.0 mL) was added dimethylamine (2 M, 1.5 mL, 2.0 equiv) dropwise, and the mixture was then stirred at 30° C. for 2 hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was triturated with ACN (3×10 mL) at 25° C. for 30 minutes to afford the title compound (1.1 g, 72% yield) as a white solid. Further purification of a 100 mg sample was carried out by preparative HPLC (column: Phenomenex luna (80 × 30 mm × 3 μm); mobile phase: 5% to 35% ACN in water (HCl)) to give the HCl salt of the title compound (50.0 mg) as a white solid. The HCl salt of the title compound (50.0 mg) was converted to the formate salt by treatment with formic acid (6.3 mg, 132 μmol, 1.0 equiv). The mixture was stirred at 25 °C for 5 minutes. The white solid was recrystallized from water and dried by lyophilization to give the formate salt of the title compound (35.0 mg, 66% yield) as a white solid.

[0134]

[0124] 1 H NMR (400 MHz, chloroform-d) δ 8.94 (s, 1H), 8.50 (s, 1H), 8.34 (s, 1H), 7.91 (dd, J = 1.6, 8.8 Hz, 1H), 7.61 (d, J = 8.8 Hz, 1H), 5.36 (q, J = 8.0 Hz, 2H), 3.92 (s, 2H), 2.47 (s, 6H). LCMS: [M+1]=343.0.

[0135] Example 2 1-[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N-methyl-N-(3-pyridylmethyl)methanamine

[0136] [ka]

[0137] Scheme 1, Step 9. 1-[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N-methyl-N-(3-pyridylmethyl)methanamine: To a solution of 4-chloro-8-(chloromethyl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole (100 mg, 299 μmol, 1.0 equiv) in ACN (5 mL) was added TEA (83 μL, 599 μmol, 2.0 equiv) and N-methyl-1-(3-pyridyl)methanamine (73.1 mg, 599 μmol, 2.0 equiv). The mixture was stirred at 50° C. for 6 hours and concentrated. The resulting residue was purified by preparative TLC (10:1 DCM:MeOH) to afford the title compound (20.0 mg, 15% yield) as a yellow solid.

[0138]

[0128] 1 H NMR (400 MHz, chloroform-d) δ 8.94 (s, 1H), 8.61 (s, 1H), 8.54 (d, J = 2.8 Hz, 1H), 8.37 (s, 1H), 7.92 - 7.74 (m, 2H), 7.57 (d, J = 8.8 Hz, 1H), 7.33 - 7.28 (m, 1H), 5.35 (q, J = 8.0 Hz, 2H), 3.78 (s, 2H), 3.64 (s, 2H), 2.26 (s, 3H).

[0129] LCMS: [M+1, M+3]=420.1, 422.2.

[0139] Example 65 1-[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N-methyl-N-(3-pyridylmethyl)methanamine

[0140] [ka]

[0141] Scheme 1, Step 9. 4-Chloro-8-[(2-methylpyrimidin-5-yl)oxymethyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole: A mixture of 4-chloro-8-(chloromethyl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole (80.0 mg, 239.4 μmol, 1.0 equiv.), 2-methylpyrimidin-5-ol (21.0 mg, 191.5 μmol, 0.8 equiv.), and CsCO (234 mg, 718 μmol, 3.0 equiv.) in acetonitrile (1.0 mL) was stirred at RT for 2 h. The mixture was poured into water (5 mL) and extracted with ethyl acetate (2 × 5 mL). The combined organic layers were dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Welch diol 150 mm x 25 mm x 5 μm; mobile phase: 5-50% ethanol in heptane) to give the title compound (15.7 mg, 15% yield) as a white solid.

[0142]

[0132] LCMS: [M+1, M+3]=408.1, 410.1.

[0133] 1 H NMR (400 MHz, DMSO-d6) δ 8.83 (s, 2H), 8.67 (s, 1H), 8.38 (s, 1H), 8.05 (d, J = 8.4 Hz, 1H), 7.87 (dd, J = 1.6, 8.8 Hz, 1H), 5.70 (q, J = 8.8 Hz, 2H), 5.04 (s, 2H), 2.70 (s, 3H). Example 66 4-chloro-8-[(4-methylimidazol-1-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole

[0143] [ka]

[0144] Scheme 1, Step 9. 4-Chloro-8-[(4-methylimidazol-1-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole: To a solution of 4-methyl-1H-imidazole (29.4 mg, 359 μmol, 0.6 equiv) in THF (2.0 mL) was added NaH (72 mg, 1.80 mmol, 3.0 equiv; 60% dispersion in oil) at 0° C. After hydrogen evolution was stopped, 4-chloro-8-(chloromethyl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole (200 mg, 599 μmol, 1.0 equiv) was added, and the mixture was allowed to warm to RT and stirred under a nitrogen atmosphere for 2 h. The reaction mixture was quenched with HO (5.0 mL), then diluted with EtOAc (6.0 mL), and extracted with EtOAc (3×3 mL). The combined organic layers were washed with brine and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Welch Ultimate XB NH2 10 x 100 x 30 mm; mobile phase: 10-15% isopropanol in heptane) to give the title compound (17.5 mg, 7.3% yield) as a white solid.

[0145]

[0136] LCMS[M+1, M+3]=380.1, 382.1.

[0137] 1 H NMR (400 MHz, chloroform-d) δ = 9.14 (s, 1H), 8.49 (d, J = 1.2 Hz, 1H), 7.89 (s, 1H), 7.84 (dd, J = 2.0, 8.4 Hz, 1H), 7.59 (d, J = 8.8 Hz, 1H), 7.16 (s, 1H), 4.84 (s, 2H), 4.70 - 4.62 (q, J = 8.0 Hz, 2H), 2.42 - 2.38 (s, 3H). Example 67 1-[[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]methyl]-4-methylpiperazin-2-one

[0146] [ka]

[0147] The title compound was prepared in a manner similar to that described for Example 66.

[0139] LCMS: [M+1, M+3]=412.1, 414.1.

[0148]

[0140] 1 H NMR (400 MHz, DMSO-d6) δ 8.94 (s, 1H), 8.18 (s, 1H), 8.03 (d, J = 8.4 Hz, 1H), 7.73 (dd, J = 1.6, 8.8 Hz, 1H), 5.74 (q, J = 8.8 Hz, 2H), 4.73 (s, 2H), 3.26 (t, J = 4.8 Hz, 2H), 3.05 (s, 2H), 2.58 (br d, J = 5.2 Hz, 2H), 2.21 (s, 3H).

[0141]

[0149] [Table 1-1]

[0150] [Table 1-2]

[0151] [Table 1-3]

[0152] [Table 1-4]

[0153] [Table 1-5]

[0154] [Table 1-6]

[0155] [Table 1-7]

[0156] [Table 1-8]

[0157] [Table 1-9]

[0158] [Table 1-10]

[0159] Example 35 [4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-(4-methylpiperazin-1-yl)methanone

[0160] [ka]

[0161] Scheme 2, Step 1. 4-Hydroxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carboxylic acid: To a solution of methyl 4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carboxylate (500.0 mg, 1.5 mmol, 1 equiv.) in THF (4 mL) and HO (1 mL) was added LiOH·HO (67.2 mg, 1.6 mmol, 1.1 equiv.), and the reaction mixture was stirred at 25° C. for 15 h. The pH of the mixture was adjusted to approximately 1 with aqueous HCl (1 M, 1 mL), and the mixture was extracted with EtOAc (3×5 mL). The combined organic extracts were dried over NaSO, filtered, and concentrated under reduced pressure to afford the title compound (500 mg, crude) as a yellow solid. This material was used in the next step without further purification.

[0162] LCMS: [M+1]=312.0. Scheme 2, Step 2. 4-Chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carbonyl chloride: A solution of 4-hydroxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carboxylic acid (50.0 mg, 160.7 μmol, 1 equiv) in SOCl (1 mL) was stirred at 60° C. for 0.5 h. The reaction mixture was concentrated under reduced pressure to give the title compound (50 mg, 89% yield) as a yellow solid. This material was used in the next step without further purification.

[0163] LCMS: [M+1] = 344.0 (the reaction was quenched with MeOH and analyzed by LCMS). Scheme 2, Step 3. [4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-(4-methylpiperazin-1-yl)methanone: To a solution of 4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carbonyl chloride (80.0 mg, 229.8 μmol, 1 equiv.) in THF (1.5 mL) was added TEA (128 μL, 919 μmol, 4 equiv.) and 1-methylpiperazine (23 μL, 207 μmol, 0.9 equiv.), and the reaction was then stirred at 25° C. for 1 h. The reaction mixture was quenched with water (1 mL) and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Waters Xbridge Prep OBD C18 150 mm × 40 mm × 10 μm; mobile phase: 25-55% ACN in water (+NH4HCO3 modifier)) to give the title compound (24.5 mg, 26% yield) as a white solid.

[0164]

[0150] 1 H NMR (400MHz, DMSO-d6) δ 8.98 (s, 1H), 8.31 (d, J = 1.6 Hz, 1H), 8.12 (d, J = 8.8 Hz, 1H), 7.88 (dd, J = 1.6, 8.8 Hz, 1H), 5.79 (q, J = 8.8 Hz, 2H), 3.55 (s, 4H), 2.35 (s, 4H), 2.21 (s, 3H).

[0151] LCMS: [M+1, M+3]=412.1, 414.1.

[0165]

[0152]

[0166] [Table 2-1]

[0167] [Table 2-2]

[0168] [Table 2-3]

[0169] Example 43 1-[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N-methyl-methanamine

[0170] [ka]

[0171] Scheme 3, Step 1. 4-Chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carbaldehyde: To a mixture of [4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]methanol (0.2 g, 634 μmol, 1 equiv.) in DCE (2 mL) was added MnO (550.8 mg, 6.34 mmol, 10 equiv.). The mixture was stirred at 80° C. for 12 h. The reaction mixture was filtered, and the solution was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (1:0 to 10:1 petroleum ether:EtOAc) to afford the title compound (0.3 g, crude) as a yellow solid. This material was used in the next step without further purification.

[0172] LCMS: [M+H]=314.0. Scheme 3, Step 2. 1-[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N-methyl-methanamine: To a solution of 4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carbaldehyde (90.0 mg, 287 μmol, 1 equiv.) in DCE (1 mL) was added methanamine (2 M, 287 μL, 2 equiv.) and AcOH (33 μL, 574 μmol, 2 equiv.), and the mixture was stirred at RT for 0.5 h. NaBH(OAc) (152 mg, 717 μmol, 2.5 equiv.) was added to the mixture, and the mixture was stirred at 25° C. for 1 h. The reaction was filtered, and the solution was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Waters Xbridge Prep OBD C18 (150 mm × 40 mm × 10 μm); mobile phase: 15-50% ACN in water (+NH4HCO3 modifier)) to give the title compound (54.6 mg, 57% yield) as a white solid.

[0173]

[0158] 1 H NMR (400MHz, chloroform-d) δ 8.92 (s, 1H), 8.34 (s, 1H), 7.79 (dd, J = 1.6, 8.8 Hz, 1H), 7.56 (d, J = 8.4 Hz, 1H), 5.35 (q, J = 8.0 Hz, 2H), 3.98 (s, 2H), 2.51 (s, 3H).

[0159] LCMS: [M+H, M+3]=329.0, 331.1.

[0174]

[0160]

[0175] [Table 3-1]

[0176] [Table 3-2]

[0177] [Table 3-3]

[0178] Table 3-4

[0179] Table 3-5

[0180] Table 3-6

[0181] Table 3-7

[0182] Table 3-8

[0183] Table 3-9

[0184] Table 3-10

[0185] Table 3-11

[0186] Table 3-12

[0187] Table 3-13

[0188] [Table 3-14]

[0189] [Table 3-15]

[0190] [Table 3-16]

[0191] Example 51 N,N-Dimethyl-1-[4-methylsulfonyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]methanamine

[0192] [ka]

[0193] To a solution of 1-[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine (30.0 mg, 87.5 μmol, 1.0 equiv.) in DMA (1.0 mL) was added NaSOMe (60.0 mg, 587.7 μmol, 6.7 equiv.). The mixture was stirred at 50° C. for 2 h. The reaction mixture was cooled to RT and purified by preparative HPLC (column: Phenomenex luna C18 80 cm × 40 mm × 3 mm; mobile phase: 15-55% ACN in water (+ formic acid modifier)) to give the title compound (4.2 mg, 12% yield) as a white solid.

[0194]

[0162] 1H NMR (400 MHz, DMSO-d6) δ 9.28 (s, 1H), 8.27 (s, 1H), 8.01 (d, J = 8.8 Hz, 1H), 7.85 (dd, J = 1.6, 8.8 Hz, 1H), 5.95 (q, J = 8.8 Hz, 2H), 3.65 (s, 3H), 3.62 (s, 2H), 2.20 (s, 6H).

[0163] LCMS [M+1] = 387.0.

[0195] Example 52 1-(4-bromo-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)-N,N-dimethylmethanamine

[0196] [ka]

[0197] A mixture of 1-[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine (30.0 mg, 87.5 μmol, 1.0 equiv) and TMSBr (79.5 μL, 612.7 μmol, 7.0 equiv) in ACN (2.0 mL) was degassed and purged with N three times, then the mixture was stirred under N atmosphere at 40° C. for 16 h. The reaction mixture was quenched by the addition of saturated aqueous NaHCO (5 mL), diluted with HO (5 mL), and extracted with EtOAc (3×5 mL). The combined organic extracts were washed with brine (3×10 mL), dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna (80 mm x 30 mm x 3 μm); mobile phase: 1-40% ACN in water (+TFA modifier)) to give the title compound (8.5 mg, 25% yield) as a white solid.

[0198]

[0165] 1H NMR (400 MHz, DMSO-d6) δ 8.90 - 8.85 (m, 1H), 8.22 (s, 1H), 8.19 - 8.15 (m, 1H), 8.03 - 7.96 (m, 1H), 7.83 - 7.76 (m, 1H), 5.84 - 5.73 (m, 2H), 3.67 (s, 2H), 2.23 (s, 6H).

[0166] LCMS: [M+1, M+3]=387.1, 389.1.

[0199] Example 53 1-[4-Fluoro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine

[0200] [ka]

[0201] To a solution of 1-[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine (50.0 mg, 146 μmol, 1.0 equiv.) in ACN (1.0 mL) was added 1,4,7,10,13,16-hexaoxacyclooctadecane (3.9 mg, 14.6 μmol, 0.1 equiv.), cesium fluoride (66.5 mg, 438 μmol, 3.0 equiv.), and tetramethylammonium chloride (1.6 mg, 14.6 μmol, 0.1 equiv.). The mixture was stirred at 60° C. for 0.5 hours. The mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Waters Xbridge Prep OBD C18 (150 × 40 mm × 10 μm); mobile phase: 30-55% ACN in water (+NH4HCO3 modifier)) to give the title compound (20.4 mg, 21% yield) as a white solid.

[0202]

[0168] 1H NMR (400 MHz, DMSO-d6) δ 8.81 (s, 1H), 8.18 (s, 1H), 7.99 (d, J = 8.8 Hz, 1H), 7.78 (d, J = 8.8 Hz, 1H), 5.52 (q, J = 9.2 Hz, 2H), 3.60 (s, 2H), 2.19 (s, 6H).

[0169] LCMS [M+1] = 327.1.

[0203] Example 54 4-Fluoro-8-((4-methylpiperazin-1-yl)methyl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole

[0204] [ka]

[0205] To a mixture of 1,4,7,10,13,16-hexaoxacyclooctadecane (10.6 mg, 40.2 μmol, 0.1 equiv.), cesium fluoride (183 mg, 1.2 mmol, 3.0 equiv.), and tetramethylammonium chloride (4.4 mg, 40 μmol, 0.1 equiv.) in ACN (4 mL) was added 4-chloro-8-[(4-methylpiperazin-1-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole (160 mg, 402 μmol, 1.0 equiv.) in one portion under a N atmosphere at 25° C. The mixture was stirred at 60° C. for 0.5 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC (Waters Xbridge BEH C18 (100 mm × 25 mm × 5 μm); mobile phase: 5-35% ACN in water (+NH4HCO3 modifier)) to give the title compound (37.0 mg, 24% yield) as a yellow solid.

[0206]

[0171] 1H NMR (400 MHz, chloroform-d) δ 8.81 (s, 1H), 8.34 (s, 1H), 7.77 (dd, J = 1.2, 8.4 Hz, 1H), 7.53 (d, J = 8.8 Hz, 1H), 5.03 (q, J = 8.2 Hz, 2H), 3.74 (s, 2H), 2.82–2.42 (m, 8H), 2.37 (s, 3H).

[0172] LCMS [M+1] = 382.4.

[0207] Example 120 4-Fluoro-8-[(2-methyl-2,7-diazaspiro[3.5]nonan-7-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole

[0208] [ka]

[0209] Step 1. 4-Chloro-8-[(2-methyl-2,7-diazaspiro[3.5]nonan-7-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole: 4-Chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carbaldehyde (250 mg, 797 μmol, 1.0 equiv.), 2-methyl-2,7-diazaspiro[3.5]nonane hydrochloride (211 mg, 1.2 mmol, 1.5 equiv.), and AcOH (91 μL, 1.5 mmol, 2 equiv.) were combined in DCE (3.0 mL) and stirred at RT for 30 min. NaBH(OAc) (507 mg, 2.3 mmol, 3.0 equiv.) was then added, and the mixture was stirred at RT for 2 h. The pH of the mixture was adjusted to pH = 9 with solid NaHCO. The reaction mixture was diluted with HO (20 mL) and extracted with ethyl acetate (2 × 25 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure to give a residue that was purified by preparative TLC (SiO, 10:1 DCM:MeOH) to give the title compound (130 mg, 37% yield) as a pale yellow solid.

[0210]

[0175] LCMS: [M+1, M+3]=438.3, 440.3. Step 2. 4-Fluoro-8-[(2-methyl-2,7-diazaspiro[3.5]nonan-7-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole: A mixture of 4-chloro-8-[(2-methyl-2,7-diazaspiro[3.5]nonan-7-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole (75.0 mg, 171.2 μmol, 1.0 equiv.), 1,4,7,10,13,16-hexaoxacyclooctadecane (4.5 mg, 17.1 μmol, 0.1 equiv.), cesium fluoride (78 mg, 514 μmol, 18.9 μL, 3.0 equiv.), and tetramethylammonium chloride (1.8 mg, 17.1 μmol, 0.1 equiv.) in acetonitrile (0.5 mL) was degassed with nitrogen, and the mixture was then stirred at 60° C. under a nitrogen atmosphere for 2 h. The reaction mixture was purified by preparative HPLC (Waters Xbridge BEH C18 column). 100 mm x 30 mm x 10 μm; mobile phase: 25-40% acetonitrile in water (+10 mM NH 4 HCO 3 )) to afford the title compound (2.0 mg, 1.3% yield) as a white solid.

[0211] LCMS: [M+1]=442.3.

[0179] 1 H NMR (400 MHz, chloroform-d) δ = 8.81 (s, 1H), 8.30 (s, 1H), 7.76 (d, J = 8.8 Hz, 1H), 7.52 (d, J = 8.4 Hz, 1H), 5.03 (q, J = 8.0 Hz, 2H), 3.66 (s, 2H), 3.17 (s, 4H), 2.43 (s, 3H), 2.39 (br s, 3H), 1.81 (m, 5H).

[0212] Example 121 N-[[4-Fluoro-8-[(4-methylpiperazin-1-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methyl]-1-methyl-pyrazole-4-carboxamide

[0213] [ka]

[0214] Example 121 was prepared in a manner similar to that described in Example 54 and Example 120. LCMS: [M+1]=519.2.

[0215]

[0182] 1 H NMR (400 MHz, DMSO-d6): δ 8.78 (t, J = 6.0 Hz, 1H), 8.17 (s, 1H), 8.13 (s, 1H), 7.96 (d, J = 8.8 Hz, 1H), 7.91 (s, 1H), 7.73 (d, J = 8.4 Hz, 1H), 5.49 (q, J = 8.8 Hz, 2H), 4.70 (d, J = 6.0 Hz, 2H), 3.87 (s, 3H), 3.65 (s, 2H), 2.49 - 2.17 (m, 8H), 2.14 (s, 3H).

[0216] Example 55 4-chloro-8-((pyridin-4-yloxy)methyl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole

[0217] [ka]

[0218] To a solution of pyridin-4-ol (8.5 mg, 89.7 μmol, 0.5 equiv.) and KCO (74.4 mg, 539 μmol, 3.0 equiv.) in DMF (0.5 mL) was added 4-chloro-8-(chloromethyl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole (60.0 mg, 178 μmol, 1.0 equiv.) at 25° C. The mixture was stirred at 25° C. for 16 h. The mixture was filtered, and the filtrate was purified by preparative HPLC (column: Phenomenex luna C18 150 mm × 25 mm × 10 μm; mobile phase: 15–45% ACN in water (+ formic acid modifier)) to give the title compound (23.7 mg, 32% yield) as a white solid.

[0219]

[0184] 1 H NMR (400 MHz, chloroform-d) δ 8.97 (s, 1H), 8.32 (s, 1H), 7.69 - 7.59 (m, 2H), 7.44 (d, J = 7.5 Hz, 2H), 6.47 (d, J = 7.6 Hz, 2H), 5.38 (q, J = 8.1 Hz, 2H), 5.17 (s, 2H).

[0185] LCMS[M+1, M+3]=393.0, 395.0.

[0220] Example 56 1-[4-chloro-2-(2,2,2-trifluoroethoxy)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine

[0221] [ka]

[0222] Step 1. 8-[(Dimethylamino)methyl]-5H-pyrimido[5,4-b]indole-2,4-diol: To a solution of 8-bromo-5H-pyrimido[5,4-b]indole-2,4-diol (2.0 g, 7.1 mmol, 1.0 equiv.) in 2-methylbutan-2-ol (24.0 mL) and HO (6.0 mL) was added CsCO (4.6 g, 14.2 mmol, 2.0 equiv.), (2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) methanesulfonate (614.4 mg, 714.0 μmol, 0.1 equiv.), and potassium [(dimethylamino)methyl]trifluoroborate (2.3 g, 14.2 mmol, 2.0 equiv.) under a nitrogen atmosphere. The mixture was stirred at 80 °C for 12 h. The reaction mixture was concentrated under reduced pressure. The resulting residue was diluted with EtOAc (100 mL) and HO (100 mL), and the aqueous layer was extracted with EtOAc (2 x 100 mL). The combined organic extracts were washed with brine (100 mL), dried over NaSO, filtered, and concentrated under reduced pressure. The resulting residue was triturated with EtOAc (5 mL) at 20 °C for 30 minutes to afford the title compound (1.1 g, 60% yield) as a white solid.

[0223]

[0188] 1 H NMR (400 MHz, DMSO-d6) δ 11.69 - 11.04 (m, 3H), 7.83 (s, 1H), 7.53 - 7.16 (m, 2H), 3.42 (s, 2H), 2.14 (s, 6H). Step 2. 1-(2,4-Dichloro-5H-pyrimido[5,4-b]indol-8-yl)-N,N-dimethyl-methanamine: To a solution of 8-[(dimethylamino)methyl]-5H-pyrimido[5,4-b]indole-2,4-diol (1.1 g, 4.2 mmol, 1.0 equiv.) was added dichlorophosphorylbenzene (8.0 mL, 57.1 mmol, 13.4 equiv.). The mixture was stirred at 140° C. for 3 h. The reaction mixture was quenched by the addition of saturated aqueous NaHCO (50 mL). The reaction mixture was concentrated under reduced pressure. The residue was diluted with EtOAc (100 mL) and HO (100 mL), and the aqueous layer was extracted with EtOAc (2×100 mL). The combined organic extracts were washed with brine (50 mL), dried over NaSO, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative TLC (SiO, 10:1 THF:MeOH) to afford the title compound (280.0 mg, crude) as a yellow oil. This material was used in the next step without further purification.

[0224]

[0191] LCMS[M+1, M+3]=295.3, 297.3. Step 3. 1-[4-chloro-2-(2,2,2-trifluoroethoxy)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine: To a solution of 1-(2,4-dichloro-5H-pyrimido[5,4-b]indol-8-yl)-N,N-dimethyl-methanamine (35.0 mg, 118.5 μmol, 1.0 equiv.) in DMF (0.5 mL) was added NaH (14.2 mg, 355.7 μmol, 60.0% purity, 3.0 equiv.) at 0° C., followed by 2,2,2-trifluoroethyl trifluoromethanesulfonate (55.0 mg, 237.1 μmol, 2.0 equiv.) at 20° C. for 2 h. LC-MS showed that 1 / 3 of the starting material remained, and one major peak with the desired mass was detected. The reaction mixture was quenched with 5 mL of HO at 0° C., extracted with EtOAc (8 mL), and the organic layer was concentrated in vacuo. The residue was purified by prep-HPLC (column: Waters Xbridge BEH C18 100 mm × 30 mm × 10 μm; mobile phase: 40-70% ACN in water (+NH4HCO3 modifier)) to give the title compound (3.1 mg, 5.8% yield) as a white solid.

[0225]

[0194] 1 H NMR (400 MHz, DMSO-d6) δ 8.09 (s, 1H), 7.92 (d, J = 8.8 Hz, 1H), 7.74 (dd, J = 1.2, 8.8 Hz, 1H), 5.46 (q, J = 8.8 Hz, 2H), 5.32 (q, J = 8.8 Hz, 2H), 3.59 (s, 2H), 2.19 (s, 6H).

[0195] LCMS[M+1, M+3]=440.8, 442.

[0226] Example 57 4-chloro-N-(4-pyridylmethyl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-amine

[0227] [ka]

[0228] Scheme 4, Step 1. 4-Chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carboxylic acid: To a solution of methyl 4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carboxylate (1.4 g, 422 μmol, 1 equiv.) in DCE (15 mL) was added hydroxy(trimethyl)stannane (6.1 g, 3,375 μmol, 8 equiv.), and the mixture was stirred at 80° C. for 12 h. The reaction mixture was quenched by the addition of an aqueous solution of FK (10 mL) and then extracted with EtOAc (3×30 mL). The combined organic extracts were washed with brine (30 mL), dried over NaSO, filtered, and concentrated under reduced pressure. The resulting residue was triturated with HCl (1 M, 50 mL), which gave a precipitate that was filtered to afford the title compound (600 mg, 43% yield) as a white solid.

[0229]

[0198] 1 H NMR (400 MHz, DMSO-d6) δ 13.15 (s, 1H), 9.00 (s, 1H), 8.84 (s, 1H), 8.36 (dd, J = 1.6, 8.8 Hz, 1H), 8.13 (d, J = 8.8 Hz, 1H), 5.79 (q, J = 8.4 Hz, 2H).

[0199] LCMS: [M+1, M+3]=330.0, 331.9.

[0230]

[0200] Example 58, Scheme 4, Step 2. tert-Butyl N-[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]carbamate: To a solution of 4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carboxylic acid (200 mg, 607 μmol, 1 equiv.) in t-BuOH (4 mL), TEA (253 μL, 182 μmol, 3 equiv.) and DPPA (250.44 mg, 910.0 μmol, 197.20 μL, 1.5 equiv.) were added. The mixture was stirred at 90°C for 1.5 h. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by preparative HPLC (Column: Waters Xbridge BEH C18 100 mm × 30 mm × 1 μm; Mobile phase: 50-70% ACN in water (+NH4HCO3 modifier)) to give the title compound (23.5 mg, 21% yield) as a white solid.

[0231]

[0202] 1 H NMR (400 MHz, DMSO-d6) δ 9.64 (s, 1H), 8.90 (s, 1H), 8.48 (s, 1H), 7.94 (d, J = 9.2 Hz, 1H), 7.83 (dd, J = 2.0, 8.8 Hz, 1H), 5.68 (q, J = 8.6 Hz, 2H), 1.52 (s, 9H).

[0203] LCMS:[M+1, M+3]=401.4, 403.4.

[0232]

[0204] Example 59, Scheme 4, Step 3. 4-Chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-amine: To a solution of tert-butyl N-[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]carbamate (80 mg, 200 μmol, 1 equiv.) in DCM (1.5 mL) was added TFA (0.5 mL). The mixture was stirred at 20° C. for 3 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex luna C18 100 mm × 40 mm × 5 μm; mobile phase: 5-35% ACN in water (+ formic acid modifier)) to give the title compound (19.8 mg, 29% yield) as a yellow solid (formate salt).

[0233]

[0206] 1 H NMR (400 MHz, DMSO-d6) δ 8.80 (s, 1H), 7.71 (d, J = 8.8 Hz, 1H), 7.37 (d, J = 2.0 Hz, 1H), 7.16 (dd, J = 2.0, 8.8 Hz, 1H), 5.60 (q, J = 8.8 Hz, 2H), 5.27 (br s, 2H).

[0207] LCMS:[M+1, M+3]=301.0, 302.8.

[0234] Scheme 4, Step 4A. 4-Chloro-N-(4-pyridylmethyl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-amine: To a solution of 4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-amine (80.0 mg, 266.0 μmol, 1 equiv.), pyridine-4-carbaldehyde (57 mg, 532 μmol, 2 equiv.), and AcOH (30 μL, 532 μmol, 2 equiv.) in DCE (1 mL) was added NaBH(OAc) (197 mg, 931 μmol, 3.5 equiv.). The mixture was stirred at 25° C. for 1 h. The pH was adjusted to approximately 9 by adding a saturated aqueous solution of NaHCO. The mixture was extracted with EtOAc (2×40 mL). The combined organic extracts were washed with water, dried over NaSO, filtered, and concentrated. The resulting residue was purified by preparative HPLC (column: Waters Xbridge BEH C18 100 mm × 30 mm × 10 μm; mobile phase: 35-55% ACN in water (+NH4HCO3 modifier)) to afford the title compound (40.0 mg, 38% yield) as a yellow solid.

[0235]

[0210] 1 H NMR (400 MHz, DMSO-d6) δ 8.77 (s, 1H), 8.50 (d, J = 6.0 Hz, 2H), 7.78 (d, J = 8.8 Hz, 1H), 7.41 (d, J = 6.0 Hz, 2H), 7.29 (dd, J = 2.0, 8.8 Hz, 1H), 7.13 (d, J = 2.4 Hz, 1H), 6.66 (t, J = 6.4 Hz, 1H), 5.60 (q, J = 8.8 Hz, 2H), 4.46 (d, J = 6.4 Hz, 2H).

[0211] LCMS: [M+1, M+3]=391.8, 393.8.

[0236] Example 60 N-[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-1-methyl-piperidine-4-carboxamide

[0237] [ka]

[0238] Scheme 4, Step 4B. N-[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-1-methyl-piperidine-4-carboxamide: To a mixture of 1-methylpiperidine-4-carboxylic acid (41.9 mg, 293 μmol, 1.1 equiv.) in DMF (1.0 mL), HATU (152 mg, 399 μmol, 1.5 equiv.), DIEA (185 μL, 1.1 mmol, 4.0 equiv.), and 4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-amine (80.0 mg, 266 μmol, 1.0 equiv.) were added, and the mixture was stirred at RT for 2 hours. The mixture was filtered to remove insoluble material and concentrated in vacuo. The resulting residue was purified by preparative HPLC (column: Waters Xbridge Prep OBD C18 150 mm × 40 mm × 10 μm; mobile phase: 15-45% ACN in water (+NH4HCO3 modifier)) to afford the title compound (22.1 mg, 19% yield) as a white solid.

[0239]

[0214] 1 H NMR (400 MHz, DMSO-d6) δ 10.14 (s, 1H), 8.91 (s, 1H), 8.70 (d, J = 1.6 Hz, 1H), 8.02 - 7.87 (m, 2H), 5.69 (q, J = 8.8 Hz, 2H), 2.88 - 2.79 (m, 2H), 2.35 - 2.28 (m, 1H), 2.16 (s, 3H), 1.88 (dt, J = 2.0, 11.2 Hz, 2H), 1.81 - 1.63 (m, 4H).

[0215] LCMS: [M+1, M+3]=425.8, 427.8.

[0240]

[0216]

[0241] [Table 4-1]

[0242] [Table 4-2]

[0243] [Table 4-3]

[0244] [Table 4-4]

[0245] Example 62 4-chloro-8-(4-methylpiperazin-1-yl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole

[0246] [ka]

[0247] Scheme 5, Step 1. Ethyl (2-cyano-4-iodophenyl)glycinate: To a solution of 2-fluoro-5-iodobenzonitrile (2.0 g, 8.1 mmol, 1.0 equiv) in NMP (20.0 mL) was added ethyl glycinate-HCl (2.3 g, 16.1 mmol, 2.0 equiv) and DIEA (4.2 mL, 24.2 mmol, 3.0 equiv) at 25° C. The mixture was stirred at 110° C. for 12 h. The mixture was cooled to 25° C. and poured into HO (50 mL). The aqueous phase was extracted with EtOAc (3×50 mL). The combined organic extracts were washed with brine, dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (100:0 to 9:1 petroleum ether:EtOAc) to afford the title compound (1.2 g, 45% yield) as a white solid.

[0248]

[0219] 1 H NMR (400 MHz, chloroform-d) δ 7.69 (d, J = 2.0 Hz, 1H), 7.64 (dd, J = 1.8, 8.8 Hz, 1H), 6.35 (d, J = 8.8 Hz, 1H), 5.21 (br s, 1H), 4.28 (q, J = 7.2 Hz, 2H), 3.96 (d, J = 5.4 Hz, 2H), 1.31 (t, J = 7.2 Hz, 3H).

[0220] LCMS [M+1] = 331.1.

[0249] Scheme 5, Step 2. 1-(tert-butyl) 2-ethyl 3-amino-5-iodo-1H-indole-1,2-dicarboxylate: A mixture of ethyl (2-cyano-4-iodophenyl)glycinate (1.0 g, 3.0 mmol, 1.0 equiv), BocO (1.3 mL, 6.0 mmol, 2.0 equiv), DMAP (37.0 mg, 303 μmol, 0.1 equiv), and TEA (422 μL, 3.0 mmol, 1.0 equiv) in DMF (10.0 mL) was degassed and purged with N three times at 25° C., then the mixture was stirred at 50° C. under a nitrogen atmosphere for 2 h. The mixture was cooled to 25° C. and poured into HO (50 mL). The aqueous phase was extracted with EtOAc (3×50 mL). The combined organic extracts were washed with brine, dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (100:0 to 10:1 petroleum ether:EtOAc) to afford the title compound (1.2 g, 92% yield) as a colorless oil.

[0250]

[0223] 1H NMR (400 MHz, chloroform-d) δ 7.96 (s, 1H), 7.89 (br d, J = 8.4 Hz, 1H), 7.38–7.28 (m, 1H), 4.24 (m, 2H), 1.63–1.49 (m, 4H), 1.44–1.39 (m, 5H), 1.33–1.24 (m, 3H). Scheme 5, Step 3. 1-(tert-butyl) 2-ethyl 3-(((dimethylamino)methylene)amino)-5-iodo-1H-indole-1,2-dicarboxylate: To a solution of 1-(tert-butyl) 2-ethyl 3-amino-5-iodo-1H-indole-1,2-dicarboxylate (800 mg, 1.86 mmol, 1.0 equiv) in DMF (8.0 mL) was added 1,1-dimethoxy-N,N-dimethylmethanamine (321 μL, 2.42 mmol, 1.3 equiv) at 25° C. The mixture was stirred at 100° C. for 16 h. The mixture was cooled to 25° C. and poured into HO (30 mL). The aqueous phase was extracted with EtOAc (3 × 50 mL). The combined organic extracts were washed with brine (3 × 30 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (100:0 to 5:1 petroleum ether:EtOAc) to afford the title compound (530 mg, 59% yield) as a yellow oil.

[0251]

[0226] 1 H NMR (400 MHz, chloroform-d) δ 8.00 (s, 1H), 7.90 (s, 1H), 7.79 (d, J = 8.8 Hz, 1H), 7.66 (dd, J = 1.8, 8.8 Hz, 1H), 4.29 (q, J = 7.0 Hz, 2H), 3.10 (br d, J = 5.6 Hz, 6H), 1.61 (s, 9H), 1.35 - 1.28 (m, 3H).

[0227] LCMS [M+1] = 486.3.

[0252] Scheme 5, Step 4. 8-Iodo-5H-pyrimido[5,4-b]indol-4-ol: To a solution of 1-(tert-butyl) 2-ethyl 3-(((dimethylamino)methylene)amino)-5-iodo-1H-indole-1,2-dicarboxylate (400 mg, 824 μmol, 1.0 equiv) in EtOH (5.0 mL) was added NH 3 HO (5.0 mL) at 25° C. The mixture was stirred at 70° C. for 16 h. The reaction mixture was filtered, and the filter cake was washed with HO (30 mL) and dried under vacuum to give the title compound (180 mg, 70% yield) as a white solid.

[0253]

[0230] 1 H NMR (400 MHz, DMSO-d6) δ 12.46 - 12.21 (m, 2H), 8.29 (d, J = 1.4 Hz, 1H), 8.01 (s, 1H), 7.71 (dd, J = 1.6, 8.6 Hz, 1H), 7.38 (d, J = 8.6 Hz, 1H). Scheme 5, Step 5. 4-Chloro-8-iodo-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole: To a solution of 4-chloro-8-iodo-5H-pyrimido[5,4-b]indole (130 mg, 395 μmol, 1.0 equiv) in DMF (2.5 mL) was added NaH (47.34 mg, 1.1 mmol, 3.0 equiv; 60% dispersion in oil) at 0° C. and stirred under a nitrogen atmosphere at 20° C. for 0.5 h. Then, 2,2,2-trifluoroethyl trifluoromethanesulfonate (183.1 mg, 789.0 μmol, 2.0 equiv) was added to the reaction mixture. The mixture was stirred under a nitrogen atmosphere at 25° C. for 16 h. The reaction mixture was quenched by the addition of a saturated aqueous solution of NH4Cl (10 mL) at 0° C. and extracted with EtOAc (3×20 mL). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative HPLC (column: Phenomenex luna C18 150 mm × 25 mm × 10 μm; mobile phase: 54% to 84% ACN in water (+ formic acid modifier)) to give the title compound (17.7 mg, 11% yield) as a white solid.

[0254]

[0233] 1 H NMR (400 MHz, chloroform-d) δ 8.95 (s, 1H), 8.76 (d, J = 1.8 Hz, 1H), 8.02 (dd, J = 1.6, 8.8 Hz, 1H), 7.38 (d, J = 8.8 Hz, 1H), 5.33 (q, J = 8.2 Hz, 2H).

[0234] LCMS[M+1, M+3]=411.8, 413.8.

[0255] Scheme 5, Step 6. 8-Iodo-4-methoxy-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole: To a solution of 4-chloro-8-iodo-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole (100 mg, 243 μmol, 1.0 equiv) in MeOH (1.0 mL) was added NaOMe (0.6 mL) at 25° C. The mixture was stirred at 25° C. for 1 h. The reaction mixture was concentrated under reduced pressure, and the residue was diluted with HO (15 mL). The aqueous phase was extracted with EtOAc (3×15 mL). The combined organic extracts were washed with brine (3×10 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to give the title compound (90.0 mg, 91% yield) as a white solid.

[0256]

[0237] 1 H NMR (400 MHz, chloroform-d) δ 8.75 - 8.67 (m, 2H), 7.91 (dd, J = 1.8, 8.8 Hz, 1H), 7.32 (d, J = 8.6 Hz, 1H), 5.14 (q, J = 8.3 Hz, 2H), 4.24 (s, 3H).

[0238] LCMS [M+1] = 408.0.

[0257] Scheme 5, Step 7. 4-Methoxy-8-(4-methylpiperazin-1-yl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole: To a solution of 8-iodo-4-methoxy-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole (80.0 mg, 196.5 μmol, 1.0 equiv.) and 1-methylpiperazine (43.5 μL, 393.0 μmol, 2.0 equiv.) in dioxane (1.0 mL) was added sodium tert-butoxide (197 μL, 2.0 equiv.; 2 M in dioxane) and Xantphos Pd G4 (18.9 mg, 19.6 μmol, 0.1 equiv.) under a nitrogen atmosphere at 25° C. The mixture was stirred at 90° C. for 1 h. The mixture was cooled to 25° C. and poured into HO (20 mL). The aqueous phase was extracted with EtOAc (3×20 mL). The combined organic extracts were washed with brine, dried over anhydrous Na.sub.2SO.sub.4, filtered, and the filtrate was concentrated under reduced pressure to give the title compound (60.0 mg, 80% yield) as a yellow solid.

[0258]

[0241] 1 H NMR (400 MHz, chloroform-d) δ 8.69 (s, 1H), 7.79 (s, 1H), 7.44–7.41 (m, 2H), 5.11 (q, J = 8.3 Hz, 2H), 4.22 (s, 3H), 3.32–3.21 (m, 4H), 2.67–2.57 (m, 4H), 2.39 (s, 3H). Scheme 5, Step 8. 4-Chloro-8-(4-methylpiperazin-1-yl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole: A solution of 4-methoxy-8-(4-methylpiperazin-1-yl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole (50.0 mg, 131.7 μmol, 1.0 equiv) in POCl (1 mL) was stirred at 110° C. for 16 h. The reaction mixture was cooled to RT and then concentrated under reduced pressure. The resulting residue was purified by preparative HPLC (column: Phenomenex Luna C18 150 mm × 25 mm × 10 μm; mobile phase: 39–89% ACN in water (+ formic acid modifier)) to afford the title compound (21.4 mg, 41% yield) as a yellow solid.

[0259]

[0244] 1 H NMR (400 MHz, chloroform-d) δ 8.90 (s, 1H), 7.87 (s, 1H), 7.55 - 7.44 (m, 2H), 5.31 (q, J = 8.2 Hz, 2H), 3.62 - 3.42 (m, 4H), 3.06 (br d, J = 4.4 Hz, 4H), 2.66 (s, 3H).

[0245] LCMS[M+1, M+3]=384.0, 386.0.

[0260]

[0246]

[0261] [Table 5-1]

[0262] [Table 5-2]

[0263] [Table 5-3]

[0264] [Table 5-4]

[0265] [Table 5-5]

[0266] Example 63 4-chloro-8-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole

[0267] [ka]

[0268] To a solution of 4-chloro-8-iodo-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole (80 mg, 194 μmol, 1.0 equiv) and 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine (52 mg, 233 μmol, 1.2 equiv) in dioxane (1 mL) and HO (0.2 mL) was added NaCO (61.8 mg, 583 μmol, 3 equiv) and Pd(dppf)Cl (14.2 mg, 19.4 μmol, 0.1 equiv) under a nitrogen atmosphere at 20° C. The mixture was stirred at 100° C. for 1 h. The mixture was cooled to RT, then poured into water (10 mL) and extracted with ethyl acetate (3 × 10 mL). The combined organic extracts were washed with brine (2 × 10 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative HPLC (column: Phenomenex luna C18 150 mm × 25 mm × 10 μm; mobile phase: 11–41% ACN in water (+ formic acid modifier)) to afford the title compound (18.6 mg, 14% yield) as a white solid.

[0269]

[0248] 1H NMR (400 MHz, methanol-d4) δ 8.88 (s, 1H), 8.44 (d, J = 1.6 Hz, 1H), 8.00 (dd, J = 1.8, 8.8 Hz, 1H), 7.87 (d, J = 9.0 Hz, 1H), 6.30 (td, J = 1.8, 3.3 Hz, 1H), 5.63 (q, J = 8.6 Hz, 2H), 3.74 (br d, J = 2.8 Hz, 2H), 3.35 (t, J = 6.0 Hz, 2H), 3.00 - 2.93 (m, 2H), 2.84 (s, 3H).

[0249] LCMS[M+1, M+3]=381.0, 383.0.

[0270] Example 64 1-[2,4-Dichloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine

[0271] [ka]

[0272] Scheme 6, Step 1. 2-(4-Bromo-2-cyano-anilino)acetamide: To a mixture of 5-bromo-2-fluorobenzonitrile (5 g, 25.0 mmol, 1 equiv.) and 2-aminoacetamide, hydrochloride (3.04 g, 27.5 mmol, 1.1 equiv.) in DMSO (50 mL) was added KCO (10.37 g, 75.0 mmol, 3 equiv.) at RT. The mixture was stirred at 100 °C for 3 h. The mixture was quenched by the addition of HO (150 mL) and extracted with EtOAc (3 × 100 mL). The combined organic extracts were washed with HO (3 × 50 mL), dried over NaSO, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (1:0 to 0:1 petroleum ether:EtOAc) to afford the title compound (2.3 g, 36% yield) as a yellow solid.

[0273]

[0252] 1 H NMR (400 MHz, DMSO-d6) δ 7.72 (d, J = 2.4 Hz, 1H), 7.56 (dd, J = 2.4, 9.2 Hz, 1H), 7.50 (s, 1H), 7.18 (s, 1H), 6.51 (d, J = 8.8 Hz, 1H), 6.39 (t, J = 5.6 Hz, 1H), 3.75 (d, J = 5.6 Hz, 2H). Scheme 6, Step 2. 3-Amino-5-bromo-1H-indole-2-carboxamide: A mixture of 2-(4-bromo-2-cyano-anilino)acetamide (5 g, 19.7 mmol, 1 equiv.) and sodium isopropoxide (1.62 g, 19.7 mmol, 1 equiv.) in IPA (50 mL) was stirred at 95° C. for 1 h. The mixture was concentrated and diluted with HO (30 mL). The pH of the mixture was adjusted to 4 with 2 N HCl and then extracted with EtOAc (15 mL). The organic layer was discarded. The pH of the aqueous layer was adjusted to pH = 8 with solid NaCO and then extracted with EtOAc (3 × 15 mL). The combined organic extracts were washed with brine (15 mL), dried over NaSO, filtered, and concentrated under reduced pressure to give the title compound (3.0 g, 53% yield) as a red solid.

[0274]

[0255] 1 H NMR (400 MHz, DMSO-d6) δ 10.39 (s, 1H), 7.93 (d, J = 1.2 Hz, 1H), 7.30 - 7.18 (m, 2H), 7.12 (s, 2H), 5.61 (s, 2H).

[0256] LCMS:[M+1, M+3]=254.0, 256.0.

[0275] Scheme 6, Step 3. 8-Bromo-5H-pyrimido[5,4-b]indole-2,4-diol: To a solution of 3-amino-5-bromo-1H-indole-2-carboxamide (4 g, 15.7 mmol, 1 equiv) in dioxane (100 mL) was added trichloromethyl chloroformate (4.52 g, 22.8 mmol, 1.45 equiv) at 25° C. The mixture was stirred at 110° C. for 2 h. The mixture was quenched by the addition of HO (40 mL). The resulting precipitate was filtered, and the solid was concentrated under reduced pressure to give the title compound (3 g, crude) as a yellow solid.

[0276]

[0259] 1 H NMR (400 MHz, DMSO-d6) δ 11.96 (s, 1H), 11.56 (s, 1H), 11.14 (s, 1H), 8.13 (d, J = 1.6 Hz, 1H), 7.46 (d, J = 1.6, 8.8 Hz, 1 H), 7.36 (d, J =8.8 Hz, 1H). Scheme 6, Step 4. 8-Bromo-2,4-dichloro-5H-pyrimido[5,4-b]indole: A mixture of 8-bromo-5H-pyrimido[5,4-b]indole-2,4-diol (1.0 g, 3.57 mmol, 1.0 equiv) in dichlorophosphorylbenzene (6.96 g, 35.7 mmol, 5.01 mL, 10.0 equiv) was degassed and purged with nitrogen three times, then the mixture was stirred at 180 °C under a nitrogen atmosphere for 3 h. The reaction mixture was quenched by the addition of a saturated aqueous solution of NaHCO (100 mL) at RT. The mixture was extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (5:1 to 1:1 petroleum ether:EtOAc) to afford the title compound (600 mg, 52% yield) as a pale yellow solid.

[0277]

[0262] 1H NMR (400 MHz, DMSO-d6) δ 12.78 (s, 1H), 8.40 (d, J = 2.0 Hz, 1H), 7.86 (dd, J = 2.0, 8.8 Hz, 1H), 7.65 (d, J = 8.8 Hz, 1H).

[0263] LCMS[M+1, M+3]=316.1, 318.1.

[0278] Scheme 6, Step 5. 8-Bromo-2,4-dichloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole: To a mixture of 8-bromo-2,4-dichloro-5H-pyrimido[5,4-b]indole (550 mg, 1.7 mmol, 1 equiv) in DMF (5.5 mL) was added NaH (174 mg, 4.3 mmol, 2.5 equiv; 60% dispersion in oil) at 0° C. The mixture was stirred at 25° C. for 0.5 h. Then, 2,2,2-trifluoroethyl trifluoromethanesulfonate (1.0 g, 4.3 mmol, 2.5 equiv) was added, and the mixture was stirred at 25° C. for 2 h. The mixture was poured into saturated aqueous NH4Cl (50 mL) and extracted with EtOAc (3 × 10 mL). The combined organic extracts were washed with brine (3 × 10 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (1:0 to 0:1 petroleum ether:EtOAc) to afford the title compound (400 mg, 58% yield) as a yellow solid.

[0279]

[0266] 1 H NMR (400 MHz, DMSO-d6) δ = 8.49 (d, J = 1.2 Hz, 1H), 8.10 - 8.01 (m, 2H), 5.75 (q, J = 8.8 Hz, 2H).

[0267] LCMS:[M+1, M+3]=398.0, 400.0.

[0280] Scheme 6, Step 6. 2-Chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-4-ol: To a mixture of 8-bromo-2,4-dichloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole (500 mg, 1.25 mmol, 1 equiv.) and potassium [(dimethylamino)methyl]trifluoroborate (414 mg, 2.51 mmol, 2 equiv.) in 2-methylbutan-2-ol (5 mL), CsCO (816.6 mg, 2.51 mmol, 2 equiv.) and [1,1′-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (81.6 mg, 125.3 μmol, 0.1 equiv.) in HO (1.3 mL) were added under a nitrogen atmosphere at 25° C. The mixture was stirred at 80° C. for 16 h. The mixture was cooled to RT, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative TLC (SiO, 4:1 DCM:MeOH) to afford the title compound (120 mg, crude) as a brown solid, which was used directly in the next step.

[0281]

[0270] LCMS:[M+1, M+3]=359.3, 361.2. Scheme 6, Step 7. 1-[2,4-Dichloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine: A solution of 2-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-4-ol (110 mg, 307 μmol, 1 equiv.) in POCl (2 mL) was stirred at 110° C. for 16 h. The mixture was cooled to RT, poured into saturated aqueous NaHCO (20 mL), and extracted with EtOAc (3 × 10 mL). The combined organic extracts were washed with brine (5 mL), dried over NaSO, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative HPLC (Column: Waters Xbridge BEH C18 100 mm × 30 mm × 10 μm; Mobile phase: 40–70% ACN in water (+NHHCO modifier)) to afford the title compound (14.2 mg, 12% yield) as a brown solid.

[0282]

[0273] 1 H NMR (400 MHz, DMSO-d) δ 8.18 (s, 1H), 8.02 (d, J = 8.4 Hz, 1H), 7.82 (d, J = 8.4 Hz, 1H), 5.72 (q, J = 8.8 Hz, 2H), 3.62 (s, 2H), 2.21 (s, 6H).

[0274] LCMS: [M+1, M+3]=376.8, 378.7.

[0283] Example 148 1-[4-chloro-5-(2,2,2-trifluoroethyl)-2-[4-(trifluoromethyl)phenyl]pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine

[0284] [ka]

[0285] Step 1. 8-[(Dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)-2-[4-(trifluoromethyl)-phenyl]pyrimido[5,4-b]indol-4-ol: To a mixture of 2-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-4-ol (100 mg, 279 μmol, 1 equiv.) and (114 mg, 418 μmol, 1.5 equiv.) in dioxane (1 mL) and HO (0.1 mL) was added KCO (116 mg, 836 μmol, 3 equiv.). The mixture was degassed with nitrogen, and Pd(PPh) (10.3 mg, 27.9 μmol, 0.1 equiv.) was added. The mixture was stirred at 100° C. for 2 h. The mixture was concentrated under reduced pressure to give a residue that was purified by preparative TLC (SiO, 10:1 ethyl acetate:methanol) to give the title compound (100 mg, crude) as a white solid.

[0286] LCMS: [M+1]=469.2. Step 2. 1-[4-chloro-5-(2,2,2-trifluoroethyl)-2-[4-(trifluoromethyl)phenyl]-pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine: A solution of 8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)-2-[4-(trifluoromethyl)phenyl]pyrimido[5,4-b]indol-4-ol (100 mg, 213.5 μmol, equiv.) in POCl3 (1 mL) was stirred at 110°C for 1 h. The residue was concentrated under reduced pressure to give a residue. The reaction mixture was cooled to RT and concentrated. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 100 mm × 30 mm × 5 μm; mobile phase: 30-60% acetonitrile in water (+0.2% formic acid)) to give the title compound (18.8 mg, 18% yield) as a white solid.

[0287]

[0280] LCMS:[M+1, M+3]=487.2, 489.2.

[0281] 1H NMR (400 MHz, DMSO-d6) δ 8.68 (d, J = 8.4 Hz, 2H), 8.37 (s, 1H), 8.04 (d, J = 8.4 Hz, 1H), 7.94 (d, J = 8.4 Hz, 2H), 7.83 (d, J = 8.4 Hz, 1H), 5.77 (q, J = 8.8 Hz, 2H), 3.76 (s, 2H), 2.31 (s, 6H). Example 149 1-(4-chloro-2-(1-methyl-1H-pyrazol-4-yl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)-N,N-dimethylmethanamine

[0288] [ka]

[0289] Scheme 7, Step 1. 8-Bromo-2,4-dichloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole: To a solution of 8-bromo-2,4-dichloro-5H-pyrimido[5,4-b]indole (27 g, 85.2 mmol, 1 equiv.) in DMF (270 mL) was added K2CO3 (23.5 g, 170 mmol, 2 equiv.) and 2,2,2-trifluoroethyl trifluoromethanesulfonate (39.5 g, 170 mmol, 2 equiv.) at RT. The mixture was stirred at RT for 4 h. The mixture was diluted with water and extracted with ethyl acetate. The combined organic extracts were combined, washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography (3:1 petroleum ether:ethyl acetate) to afford the title compound (28 g, 82% yield) as a yellow solid.

[0290]

[0284] LCMS:[M+1, M+3]=398.1, 400.1. Scheme 7, Step 2. 8-Bromo-2-chloro-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole: To a solution of 8-bromo-2,4-dichloro-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole in methanol (50 mL) was added NaOMe (1.13 g, 6.27 mmol, 1 equivalent; 30% solution in methanol). The mixture was heated to 60° C. for 30 minutes. The reaction was cooled to RT, diluted with water, and the mixture was extracted with ethyl acetate. The combined organic layers were washed with brine (5 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to give the title compound (5.1 g, crude) as a pale yellow solid.

[0291]

[0287] LCMS:[M+1, M+3]=393.9, 395.9. Step 3. 1-[2-chloro-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine: To a mixture of 8-bromo-2-chloro-4-methoxy-5-(2,2,2-trifluoroethyl)-pyrimido[5,4-b]indole (4.6 g, 11.7 mmol, 1 equiv.) and potassium ((dimethylamino)methyl)trifluoroborate (2.31 g, 14.0 mmol, 1.2 equiv.) in 2-methylbutan-2-ol (46 mL), CsCO (7.60 g, 23.3 mmol, 2 equiv.) and [1,1′-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (760 mg, 1.17 mmol, 0.1 equiv.) in HO (10 mL) were added at RT under a nitrogen atmosphere. The mixture was stirred at 80° C. for 5 h. The mixture was cooled to RT and extracted with EtOAc (3×10 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography (0-100% ethyl acetate in petroleum ether) to give the title compound (3.2 g, 68% yield) as a peach-colored solid.

[0292]

[0290] LCMS:[M+1, M+3]=373.1, 375.1. Scheme 7, Step 4. 1-[4-Methoxy-2-(1-methylpyrazol-4-yl)-5-(2,2,2-trifluoroethyl)-2,3-dihydropyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine: To a mixture of 1-[2-chloro-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine (200 mg, 537 μmol, 1 equiv.) and 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole (167 mg, 805 μmol, 1.5 equiv.) in HO (0.2 mL) and dioxane (2 mL), Pd(PPh) (62.0 mg, 53.7 μmol, 0.1 equiv.) and KCO (222 mg, 1.61 mmol, 3 equiv.) were added under a nitrogen atmosphere at RT. The mixture was stirred at 100° C. for 12 h. The mixture was cooled to RT, water (1 mL) was added, and the mixture was extracted with EtOAc (3 × 1 mL). The combined organic layers were washed with brine, dried over NaSO, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO, 10:1 ethyl acetate:methanol) to give the title compound (120 mg, 53% yield) as a yellow solid.

[0293] LCMS: [M+1]=419.3.

[0294] Scheme 7, Step 5. 1-(4-chloro-2-(1-methyl-1H-pyrazol-4-yl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)-N,N-dimethylmethanamine:

[0295] A mixture of 1-[4-methoxy-2-(1-methylpyrazol-4-yl)-5-(2,2,2-trifluoroethyl)-2,3-dihydropyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine (100 mg, 238 μmol, 1 equiv.) in dichlorophosphorylbenzene (1 mL, 7.14 mmol, 30.0 equiv.) was stirred at 140° C. for 2 h. The mixture was cooled to RT, poured into saturated aqueous NaHCO (5 mL), and extracted with EtOAc (3×2 mL). The combined organic layers were washed with brine, dried over NaSO, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 100 mm × 30 mm × 5 μm; mobile phase: 15-45% acetonitrile in water (+0.2% formic acid)) to afford the title compound (49.4 mg, 49% yield) as a white solid.

[0294]

[0296] LCMS: [M+1, M+3]=423.2, 425.3.

[0297] 1 H NMR (400 MHz, DMSO-d6) δ 8.47 (s, 1H), 8.19 (s, 1H), 8.10 (s, 1H), 7.95 (d, J = 8.4 Hz, 1H), 7.74 (d, J = 8.0 Hz, 1H), 5.68 (q, J = 8.8 Hz, 2H), 3.93 (s, 3H), 3.60 (s, 2H), 2.20 (s, 6H).

[0298]

[0295] [Table 6]

[0296] Example 152 4-chloro-2-methyl-8-[(4-methylpiperazin-1-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole

[0297] [ka]

[0298]

[0299] Step 1. 2-Chloro-4-methoxy-5-(2,2,2-trifluoroethyl)-8-vinyl-pyrimido[5,4-b]indole:

[0300] To a mixture of 8-bromo-2-chloro-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole (2.0 g, 5.0 mmol, 1.0 equiv.) and 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (859 mg, 5.58 mmol, 1.1 equiv.) in 2-methylbutan-2-ol (32 mL) and HO (8 mL), CsCO (1.82 g, 5.58 mmol, 1.1 equiv.) and [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (330 mg, 507 μmol, 0.1 equiv.) were added under a nitrogen atmosphere at RT. The mixture was stirred at 80 °C for 1 h. The reaction mixture was cooled to RT and extracted with CHCl (3 × 20 mL). The combined organic layers were washed with brine, dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (15-50% ethyl acetate in petroleum ether) to afford the title compound (540 mg, 31% yield) as a white solid.

[0299]

[0301] LCMS: [M+1, M+3]=342.0, 344.0.

[0302] 1 H NMR (400 MHz, chloroform-d) δ 8.31 (s, 1H), 7.74 (dd, J = 1.6, 8.8 Hz, 1H), 7.43 (d, J = 8.8 Hz, 1H), 6.86 (dd, J = 10.8, 17.6 Hz, 1H), 5.83 (d, J = 17.6 Hz, 1H), 5.31 (d, J = 10.8 Hz, 1H), 5.08 (q, J = 8.4 Hz, 2H),4.24 (s, 3H).

[0303] Step 2. 4-Methoxy-2-methyl-5-(2,2,2-trifluoroethyl)-8-vinyl-pyrimido[5,4-b]indole:

[0304] To a mixture of 2-chloro-4-methoxy-5-(2,2,2-trifluoroethyl)-8-vinyl-pyrimido[5,4-b]indole (540 mg, 1.58 mmol, 1.0 equiv.) and 2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane (397 mg, 3.16 mmol, 2.0 equiv.) in toluene (5.0 mL) and HO (1.0 mL), Pd(OAc) (71 mg, 316 μmol, 0.2 equiv.), tricyclohexylphosphine (89 mg, 316 μmol, 0.2 equiv.), and KPO (1.68 g, 7.90 mmol, 5.0 equiv.) were added under a nitrogen atmosphere at RT. The mixture was stirred at 120 °C for 16 h. The reaction mixture was cooled to RT and extracted with CHCl (3 × 25 ml). The combined organic layers were washed with brine, dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (3–5% ethyl acetate in petroleum ether) to afford the title compound (170 mg, 33% yield) as a green solid.

[0300]

[0305] LCMS: [M+1]=322.1.

[0306] Step 3. 4-Methoxy-2-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carbaldehyde:

[0307] To a mixture of 4-methoxy-2-methyl-5-(2,2,2-trifluoroethyl)-8-vinyl-pyrimido[5,4-b]indole (170 mg, 529 μmol, 1.0 equiv) in dioxane (2.0 mL) and HO (1.0 mL), KOsO·2HO (4.9 mg, 13.2 μmol, 0.025 equiv), NaIO (566 mg, 2.65 mmol, 5.0 equiv), and 2,6-dimethylpyridine (185 μL, 1.59 mmol, 3.0 equiv) were added under a nitrogen atmosphere at RT. The mixture was stirred at RT for 1 h. The reaction mixture was poured into HO (5 mL) and extracted with ethyl acetate (3 × 5 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was washed with ethyl acetate to give the title compound (150 mg, 88% yield) as a white solid.

[0301]

[0308] LCMS: [M+1]=324.2.

[0309] 1 H NMR (400 MHz, chloroform-d) δ 10.13 (s, 1H), 8.85 (d, J = 1.2 Hz, 1H), 8.20 (dd, J = 1.6, 8.8 Hz, 1H), 7.59 (d, J = 8.8 Hz, 1H), 5.18 (q, J = 8.4 Hz, 2H), 4.23 (s, 3H), 2.84 - 2.77 (m, 3H).

[0310] Step 4. 4-Methoxy-2-methyl-8-[(4-methylpiperazin-1-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole:

[0311] To a mixture of 4-methoxy-2-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carbaldehyde (100 mg, 309 μmol, 1.0 equiv.) and 1-methylpiperazine (69 μL, 619 μmol, 2.0 equiv.) in DCE (1.0 mL) was added AcOH (17.7 μL, 309 μmol, 1.0 equiv.) at RT. NaBH(OAc) (196.6 mg, 928.0 μmol, 3.0 equiv.) was then added, and the mixture was stirred at RT for 2 h. Water (10 mL) was added dropwise to the reaction mixture, and the pH was adjusted to pH = 8 with a saturated aqueous solution of NaHCO. The mixture was extracted with ethyl acetate (2 × 10 mL). The combined organic extracts were dried over Na.sub.2SO.sub.4, filtered, and concentrated to give the title compound (100 mg, 79% yield) as a yellow solid.

[0302]

[0312] LCMS: [M+1]=408.2.

[0313] 1 H NMR (400 MHz, chloroform-d) δ 8.25 (s, 1H), 7.67–7.61 (m, 1H), 7.44 (d, J = 8.8 Hz, 1H), 5.17–5.06 (m, 2H), 4.20 (s, 3H), 3.68 (s, 2H), 2.77 (s, 3H), 2.62–2.43 (m, 8H), 2.30 (s, 3H).

[0314] Step 5. 4-chloro-2-methyl-8-[(4-methylpiperazin-1-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole:

[0315] POCl3 (1.0 mL) was added in one portion to a flask containing 4-methoxy-2-methyl-8-[(4-methylpiperazin-1-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole (80 mg, 196 μmol, 1.0 equiv.) under a nitrogen atmosphere at RT. The mixture was stirred at 120 °C for 18 h. The mixture was cooled to RT and quenched by the addition of saturated aqueous NaHCO3 (1 mL). The mixture was extracted with ethyl acetate (3 × 10 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 75 mm x 30 mm x 3 μm; mobile phase: 20-45% acetonitrile in water (+0.04% HCl)) to give the title compound (10.5 mg) as a white solid.

[0303]

[0316] LCMS: [M+1, M+3]=412.2, 414.2.

[0317] 1 H NMR (400 MHz, DMSO-d6) δ 8.55 (s, 1H), 8.14 - 7.96 (m, 2H), 5.74 (q, J = 8.0 Hz, 2H), 4.75 - 4.44 (m, 2H), 3.72 - 3.17 (m, 8H), 2.90 - 2.73 (m, 6H).

[0304] Example 153 1-[4-chloro-2-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N-methyl-methanamine

[0305] [ka]

[0306]

[0318] Step 1. 4-Chloro-2-methyl-5-(2,2,2-trifluoroethyl)-8-vinyl-pyrimido[5,4-b]indole:

[0319] Dichlorophosphoryloxybenzene (10 mL) was added to a flask containing 4-methoxy-2-methyl-5-(2,2,2-trifluoroethyl)-8-vinyl-pyrimido[5,4-b]indole (800 mg, 2.49 mmol, 1.0 equiv.), and the mixture was stirred at 110° C. for 2 h. The reaction mixture was cooled to RT and quenched by the addition of a saturated aqueous solution of NaHCO. The mixture was extracted with ethyl acetate (3×50 mL). The combined organic layers were washed with brine, dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to give the title compound (800 mg, crude) as a brown solid.

[0307]

[0320] LCMS: [M+1]=326.0.

[0321] Step 2. 4-Chloro-2-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carbaldehyde:

[0322] To a mixture of 4-chloro-2-methyl-5-(2,2,2-trifluoroethyl)-8-vinyl-pyrimido[5,4-b]indole (800 mg, 2.46 mmol, 1.0 equiv) in dioxane (8.0 mL) and HO (4.0 mL), KOsO·2HO (22.6 mg, 61.4 μmol, 0.025 equiv), NaIO (2.63 g, 12.28 mmol, 5.0 equiv), and 2,6-dimethylpyridine (859 μL, 7.37 mmol, 3.0 equiv) were added under a nitrogen atmosphere at RT. The reaction was stirred at RT for 1 h. The reaction mixture was poured into water (5 mL) and extracted with ethyl acetate (3 × 5 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (10-20% ethyl acetate in petroleum ether) to give the title compound (350 mg, 43% yield) as a white solid.

[0308]

[0323] LCMS: [M+1]=326.0.

[0324] Step 3. 1-[4-chloro-2-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N-methyl-methanamine:

[0325] To a mixture of 4-chloro-2-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carbaldehyde (40.0 mg, 122 μmol, 1.0 equiv.) and methylamine (122.0 μL, 244 μmol, 2.0 equiv.; 2 M in THF) in DCM (0.4 mL) was added AcOH (21 μL, 366 μmol, 3.0 equiv.). The mixture was stirred at RT for 30 min, then NaBH(OAc) (78 mg, 366 μmol, 3 equiv.) was added and the reaction was stirred at RT for 2 h. The reaction was diluted with water and DCM, and the layers were separated. The organic layer was dried over NaSO, filtered, and concentrated. The residue was purified by preparative HPLC (column: Phenomenex luna C18 80 mm × 30 mm × 3 μm; mobile phase: 5 to 40% acetonitrile in water (0.04% HCl)) to give the title compound (12.2 mg, 29% yield; hydrochloride salt) as a white solid.

[0309]

[0326] LCMS: [M+1, M+3]=343.2, 345.2.

[0327] 1 H NMR (400 MHz, DMSO-d6) δ 9.04 (br s, 2H), 8.50 (d, J = 1.2 Hz, 1H), 8.12 - 8.07 (m, 1H), 7.93 (dd, J = 1.2, 8.8 Hz, 1H), 5.74 (q, J = 8.8 Hz, 2H), 4.34 (s, 2H), 2.78 (s, 3H), 2.59 (s, 3H).

[0310] Example 154 4-chloro-8-(3,8-diazabicyclo[3.2.1]octan-3-ylmethyl)-2-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole

[0311] [ka]

[0312]

[0328] Step 1. tert-Butyl 3-[[4-chloro-2-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]methyl]-3,8-diazabicyclo[3.2.1]octane-8-carboxylate:

[0329] To a mixture of 4-chloro-2-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carbaldehyde (80 mg, 244 μmol, 1 equiv.) and tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate (104 mg, 488 μmol, 2 equiv.) in DMF (1 mL) was added TMSCl (186 μL, 1.46 mmol, 6 equiv.). The mixture was stirred at RT for 20 min, after which NaBH(OAc) (155 mg, 732 μmol, 3 equiv.) was added. The reaction was then stirred at RT for 2 h. The reaction mixture was quenched with HO (1 mL) and extracted with EtOAc (3 × 1 mL). The combined organic layers were dried over NaSO and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO 2 , 1:1 ethyl acetate:petroleum ether) to afford the title compound (80 mg, 61% yield) as a yellow oil.

[0313]

[0330] LCMS: [M+1, M+3]=524.3, 526.3.

[0331] Step 2. 4-chloro-8-(3,8-diazabicyclo[3.2.1]octan-3-ylmethyl)-2-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole:

[0332] A solution of tert-butyl 3-[[4-chloro-2-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]methyl]-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (100 mg, 191 μmol, 1 equiv.) in HCl / dioxane (1 mL) was stirred at RT for 1 h. The mixture was concentrated under reduced pressure to give the title compound (80 mg, 89% yield; hydrochloride salt) as a white solid.

[0314]

[0333] LCMS: [M+1, M+3]=424.2, 426.2.

[0334] 1 H NMR (400 MHz, DMSO-d6) δ 9.64 (br s, 1H), 8.51 - 8.48 (m, 1H), 8.23 ​​- 7.87 (m, 2H), 5.74 (q, J = 8.8 Hz, 2H), 4.75 (br s, 4H), 4.56 - 4.28 (m, 2H), 4.14 (s, 2H), 2.78 (s, 3H), 2.45 - 2.26 (m, 2H), 2.10 - 1.91 (m, 2H). Example 155 4-chloro-8-(3,6-diazabicyclo[3.1.1]heptan-3-ylmethyl)-2-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole

[0315] [ka]

[0316]

[0335] Example 155 was prepared in a manner similar to that described for Example 154.

[0336] LCMS: [M+1, M+3]=410.1, 412.1.

[0317]

[0337] 1H NMR (400 MHz, DMSO-d6) δ 12.12 (br s, 1H), 9.90 (br s, 1H), 9.25 (br s, 1H), 8.62 - 7.70 (m, 3H), 5.86 - 5.61 (m, 2H), 4.96 - 4.60 (m, 1H), 4.38 - 4.22 (m, 2H), 4.19 - 3.93 (m, 2H), 3.88 - 3.62 (m, 2H), 3.22 - 3.01 (m, 1H), 2.77 (s, 3H).

[0318] Example 156 N-((4-chloro-8-((dimethylamino)methyl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-2-yl)methyl)-1-methyl-1H-pyrazole-4-carboxamide

[0319] [ka]

[0320]

[0338] Scheme 8, Step 1. tert-Butyl N-[2-[(5-bromo-2-carbamoyl-1H-indol-3-yl)amino]-2-oxo-ethyl]carbamate:

[0339] To a solution of 2-(tert-butoxycarbonylamino)acetic acid (6.1 g, 35.2 mmol, 1.5 equiv.) in DMF (40.0 mL) was added TEA (9.5 g, 94.0 mmol, 13.1 mL, 4.0 equiv.), EDCI (9.0 g, 47.0 mmol, 2.0 equiv.), and 1-oxidopyridin-1-ium-2-ol (5.2 g, 47.0 mmol, 2.0 equiv.). 3-Amino-5-bromo-1H-indole-2-carboxamide (5.9 g, 23.5 mmol, 1.0 equiv.) was then added, and the mixture was stirred at 30 °C for 2 hours. The reaction mixture was quenched with HO (100 mL). The resulting suspension was filtered through a pad of Celite, and the filter cake was washed with ethyl acetate to give the crude product. The filtrate was extracted with ethyl acetate (100 mL × 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was triturated with ethyl acetate at RT for 20 min to give the title compound (6.1 g, 58% yield) as a brown solid.

[0321]

[0340] LCMS: [M+1, M+3]=411.2, 413.2.

[0341] 1 H NMR (400 MHz, DMSO-d6) δ 11.63 (s, 1H), 9.94 (s, 1H), 7.83 (s, 1H), 7.73 (br s, 1H), 7.40 - 7.22 (m, 4H), 3.78 (d, J = 6.0 Hz, 2H), 1.42 (s, 9H).

[0342] Scheme 8, Step 2. tert-Butyl N-[(8-bromo-4-hydroxy-5H-pyrimido[5,4-b]indol-2-yl)methyl]carbamate:

[0343] To a solution of tert-butyl N-[2-[(5-bromo-2-carbamoyl-1H-indol-3-yl)amino]-2-oxo-ethyl]carbamate (4.0 g, 9.8 mmol, 1.0 equiv) in EtOH (50 mL) and HO (2.5 mL) was added NaOH (8.0 M, 2.5 mL, 2.0 equiv) at 0 °C. The mixture was stirred at RT for 16 h. The reaction mixture was quenched with water (80 mL). The suspension was then filtered through a pad of Celite, and the filter cake was washed with ethyl acetate. The filtrate was extracted with EtOAc (150 mL × 3). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was triturated with ethyl acetate at RT for 20 min to give the title compound (3.2 g, 76% yield) as a brown solid.

[0322]

[0344] LCMS: [M+1, M+3]=393.1, 395.1.

[0345] 1 H NMR (400 MHz, DMSO-d6) δ 12.20 (br s, 2H), 8.05 (d, J = 1.6 Hz, 1H), 7.55 (dd, J = 1.6, 8.8 Hz, 1H), 7.48 (d, J = 8.8 Hz, 1H), 7.20 (br t, J = 5.4 Hz, 1H), 4.16 (d, J = 5.6 Hz, 2H), 1.42 (s, 9H).

[0346] Scheme 8, Step 3. (8-Bromo-4-chloro-5H-pyrimido[5,4-b]indol-2-yl)methanamine hydrochloride:

[0347] A solution of tert-butyl N-[(8-bromo-4-hydroxy-5H-pyrimido[5,4-b]indol-2-yl)methyl]carbamate (4.0 g, 10.1 mmol, 1.0 equiv) in POCl (20.0 mL) was stirred at 100° C. for 16 h. The mixture was cooled to RT, and the solvent was removed under reduced pressure. The residue was dissolved in THF and quenched by the addition of aqueous NaHCO at RT. Once the pH was determined to be 7, the mixture was extracted with ethyl acetate (3×10 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure to give the title compound (3.1 g) as a black solid.

[0323]

[0348] LCMS: [M+1, M+3]=311.0, 312.9.

[0349] Scheme 8, Step 4. tert-Butyl N-[(8-bromo-4-chloro-5H-pyrimido[5,4-b]indol-2-yl)methyl]carbamate:

[0350] To a solution of (8-bromo-4-chloro-5H-pyrimido[5,4-b]indol-2-yl)methanamine (3.4 g, 10.9 mmol, 1.0 equiv.) in THF (20 mL) and HO (20 mL) was added NaHCO (1.8 g, 21.8 mmol, 2.0 equiv.) and BocO (1.2 g, 5.4 mmol, 0.5 equiv.). The mixture was stirred at RT for 2 h. The reaction mixture was diluted with HO (20 mL) and extracted with ethyl acetate (3 × 20 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0–75% ethyl acetate in petroleum ether) to afford the title compound (2.9 g, 55% yield) as a brown solid. LCMS: [M+1, M+3]=411.2, 413.2.

[0324]

[0351] 1H NMR (400 MHz, DMSO-d6) δ 12.48 (s, 1H), 8.34 (d, J = 1.6 Hz, 1H), 7.82 (dd, J = 2.0, 8.8 Hz, 1H), 7.64 (d, J = 8.8 Hz, 1H), 7.38 (t, J = 6.2 Hz, 1H), 4.45 (d, J = 6.0 Hz, 2H), 1.43 (s, 9H).

[0352] Scheme 8, Step 5. tert-Butyl N-[[8-bromo-4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methyl]carbamate:

[0353] To a 100 mL three-neck flask was added a solution of tert-butyl N-[(8-bromo-4-chloro-5H-pyrimido[5,4-b]indol-2-yl)methyl]carbamate (2.1 g, 5.1 mmol, 1.0 equiv.) in DMF (20 mL). The solution was degassed with nitrogen and then cooled to 0 °C. NaH (306.0 mg, 7.6 mmol, 60% purity, 1.5 equiv.) was added to the mixture in 5 portions under a nitrogen atmosphere at 0 °C and stirred at 0 °C for 1 h. A solution of 2,2,2-trifluoroethyl trifluoromethanesulfonate (1.8 g, 7.6 mmol, 1.5 equiv.) in DMF (2.0 mL) was added dropwise to the mixture, and the resulting mixture was stirred at 0 °C for 1 h. The mixture was quenched by the addition of a saturated aqueous solution of NH4Cl (50 mL). The mixture was extracted with ethyl acetate (3 × 30 mL), and the combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (50:1 to 10:1 petroleum ether:ethyl acetate) to afford the title compound (1.9 g, 76% yield) as a white solid.

[0325]

[0354] LCMS: [M+1, M+3]=493.2, 495.2.

[0355] 1H NMR (400 MHz, chloroform-d) δ 8.54 (d, J = 1.6 Hz, 1H), 7.84 (dd, J = 2.0, 8.8 Hz, 1H), 7.46 (d, J = 8.8 Hz, 1H), 5.73 (br s, 1H), 5.31 (q, J = 8.0 Hz, 2H), 4.73 (d, J = 4.8 Hz, 2H), 1.52 (s, 9H).

[0356] Scheme 8, Step 6. tert-Butyl N-[[8-[(dimethylamino)methyl]-4-hydroxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methyl]carbamate.

[0326]

[0357] 2-(2-methyl-3-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methyl)carbamate (600 mg, 1.2 mmol, 1.0 equiv.), potassium ((dimethylamino)methyl)trifluoroborate (401 mg, 2.4 mmol, 2.0 equiv.), and CsCO (792 mg, 2.4 mmol, 2.0 equiv.). To a mixture of methylbutan-2-ol (5.0 mL) and HO (1.25 mL), dicyclohexyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane; methanesulfonate; [2-[2-(methylamino)phenyl]phenyl]palladium(II) (104.5 mg, 121.5 μmol, 0.1 equiv.) was added in one portion at RT under a nitrogen atmosphere. The mixture was stirred at 80 °C for 16 h. The reaction mixture was poured into water (10 mL) and extracted with EtOAc (2 × 10 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (50:1 to 10:1 petroleum ether:ethyl acetate) to afford the title compound (350 mg, 64% yield) as a white solid.

[0327]

[0358] LCMS: [M+1]=454.4.

[0359] 1 H NMR (400 MHz, methanol-d4) δ 8.20 (s, 1H), 7.76 (d, J = 8.8 Hz, 1H), 7.64 (d, J = 8.8 Hz, 1H), 5.55 (q, J = 8.8 Hz, 2H), 4.32 (s, 2H), 4.07 (s, 2H), 2.59 (s, 6H), 1.49 (s, 9H).

[0360] Scheme 8, Step 7. [4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methanamine hydrochloride:

[0361] A mixture of tert-butyl N-[[8-[(dimethylamino)methyl]-4-hydroxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methyl]carbamate (40.0 mg, 88.2 μmol, 1 equiv.) in POCl3 (0.3 mL) was degassed with nitrogen, and the mixture was stirred at 60 °C under a nitrogen atmosphere for 16 h. The mixture was cooled to RT and then poured into HO (5 mL). The mixture was stirred for 2 min and then concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 75 mm × 0 mm × 3 μm; mobile phase: 5–30% acetonitrile in water (0.04% HCl)) to give the title compound (20.0 mg, 55% yield) as a white solid.

[0328]

[0362] LCMS: [M+1]=372.1, 374.0.

[0363] 1H NMR (400 MHz, DMSO-d6) δ 11.16 (br s, 1H), 8.70 (br s, 3H), 8.53 (s, 1H), 8.18 (d, J = 8.8 Hz, 1H), 8.12 (d, J = 8.8 Hz, 1H), 5.82 (q, J = 8.4 Hz, 2H), 4.52 (d, J = 5.2 Hz, 2H), 4.45 (q, J = 5.6 Hz, 2H), 2.72 (d, J = 4.8 Hz, 6H).

[0364] Scheme 8, Step 8. N-((4-chloro-8-((dimethylamino)methyl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-2-yl)methyl)-1-methyl-1H-pyrazole-4-carboxamide:

[0365] [4-Chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methanamine (200 mg, 538 μmol, 1.0 equiv.) and 1-methylpyrazole-4-carbonyl chloride (156 mg, 1.1 mmol, 2.0 equiv.) were combined in pyridine (2 mL), and the mixture was stirred at RT under a nitrogen atmosphere for 30 minutes. The reaction mixture was poured into water (20 mL). The aqueous phase was extracted with ethyl acetate (2 × 20 mL). The organic extract was concentrated, and the residue was purified by preparative HPLC (column: Waters Xbridge C18 150 mm × 50 mm × 10 μm; mobile phase: 15–45% acetonitrile in H2O (10 mM NH4HCO3)) to give the title compound (25.4 mg, 9.2% yield) as a white solid.

[0329]

[0366] LCMS: [M+1, M+3]=480.1, 482.2.

[0367] 1H NMR (400 MHz, DMSO-d6) δ 8.80 (t, J = 5.8 Hz, 1H), 8.17 (s, 1H), 8.16 (s, 1H), 7.98 (d, J = 8.8 Hz, 1H), 7.91 (s, 1H), 7.81 - 7.72 (dd, J = 1.2, 8.4 Hz, 1H), 5.71 (q, J = 8.4 Hz, 2H), 4.72 (d, J = 6.0 Hz, 2H), 3.87 (s, 3H), 3.63 (s, 2H), 2.21 (s, 6H).

[0368]

[0330] [Table 7-1]

[0331] [Table 7-2]

[0332] [Table 7-3]

[0333] Example 167 N-[[4-chloro-8-[(4-methylpiperazin-1-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methyl]-1-methyl-pyrazole-4-carboxamide

[0334] [ka]

[0335]

[0369] Step 1. tert-Butyl N-[[4-chloro-5-(2,2,2-trifluoroethyl)-8-vinyl-pyrimido[5,4-b]indol-2-yl]methyl]carbamate:

[0370] A mixture of 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (1.1 g, 7.2 mmol, 1.2 mL, 1.2 equiv.), tert-butyl N-[[8-bromo-4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methyl]carbamate (3.0 g, 6.0 mmol, 1.0 equiv.), CsCO (2.1 g, 6.6 mmol, 1.1 equiv.), [1,1′-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (396 mg, 608 μmol, 0.1 equiv.) in 2-methylbutan-2-ol (120 mL) and HO (30 mL) was degassed with nitrogen, and the mixture was then stirred at 80° C. under a nitrogen atmosphere for 2 h. The reaction mixture was cooled to RT, then poured into HO (50 mL) and extracted with ethyl acetate (3 × 30 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (1–10% ethyl acetate in petroleum ether) to afford the title compound (2.8 g, crude) as a white solid.

[0336]

[0371] LCMS[M+1, M+3]=441.2, 443.2.

[0372] 1 H NMR (400 MHz, chloroform-d) δ = 8.40 (s, 1H), 7.85 (dd, J = 1.6, 8.8 Hz, 1H), 7.52 (d, J = 8.8 Hz, 1H), 6.91 (dd, J = 10.8, 17.6 Hz, 1H), 5.89 (d, J = 18.0 Hz, 1H), 5.76 (br s, 1H), 5.38 - 5.28 (m, 3H), 4.74 (br d, J = 4.8 Hz, 2H), 1.53 (s, 9H).

[0373] Step 2. tert-Butyl N-[[4-chloro-8-formyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methyl]carbamate.

[0337]

[0374] To a solution of tert-butyl N-[[4-chloro-5-(2,2,2-trifluoroethyl)-8-vinyl-pyrimido[5,4-b]indol-2-yl]methyl]carbamate (2.2 g, 4.9 mmol, 1.0 equiv.) in 1,4-dioxane (80 mL) and HO (40 mL), KOsO·HO (45.9 mg, 124.7 μmol, 0.025 equiv.), NaIO (4.2 g, 19.9 mmol, 4.0 equiv.), and 2,6-dimethylpyridine (1.1 mL, 9.9 mmol, 2.0 equiv.) were added, and the mixture was stirred at RT for 3 h. The reaction mixture was poured into HO (50 mL) and extracted with ethyl acetate (3 × 50 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (1-50% ethyl acetate in petroleum ether) to give the title compound (2.5 g, crude) as a white solid.

[0338]

[0375] LCMS[M+1-56, M+3-56]=387.0, 389.0.

[0376] 1 H NMR (400 MHz, chloroform-d) δ = 10.19 (s, 1H), 8.93 (s, 1H), 8.32 (dd, J = 0.8, 8.4 Hz, 1H), 7.70 (d, J = 8.4 Hz, 1H), 5.73 (br s, 1H), 5.39 (q, J = 7.6 Hz, 2H), 4.76 (br d, J = 3.6 Hz, 2H), 1.53 (s, 9H).

[0377] Step 3. tert-Butyl N-[[4-chloro-8-[(4-methylpiperazin-1-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methyl]carbamate:

[0378] To a solution of tert-butyl N-[[4-chloro-8-formyl-5-(2,2,2-trifluoroethyl)-pyrimido[5,4-b]indol-2-yl]methyl]carbamate (500 mg, 1.1 mmol, 1.0 equiv.) and 1-methylpiperazine (226.1 mg, 2.2 mmol, 250.4 μL, 2.0 equiv.) in DCE (5.0 mL) was added AcOH (129 μL, 2.2 mmol, 2.0 equiv.) and NaBH(OAc) (598 mg, 2.8 mmol, 2.5 equiv.), and the mixture was stirred at RT for 1 h. The reaction mixture was poured into saturated aqueous NaHCO (20 mL) and extracted with ethyl acetate (3 × 5 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (10:1 DCM:MeOH) to afford the title compound (1.1 g, 92% yield) as a white solid.

[0339]

[0379] LCMS[M+1, M+3]=527.4, 529.3.

[0380] 1 H NMR (400 MHz, chloroform-d) δ = 8.33 (s, 1H), 7.76 (d, J = 8.8 Hz, 1H), 7.52 (d, J = 8.4 Hz, 1H), 5.78 (br s, 1H), 5.32 (q, J = 8.0 Hz, 2H), 4.73 (d, J = 4.8 Hz, 2H), 3.72 (s, 2H), 2.56 (br s, 8H), 2.34 (s, 3H), 1.53 (s, 9H).

[0381] Step 4. [4-chloro-8-[(4-methylpiperazin-1-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methanamine hydrochloride:

[0382] To a solution of tert-butyl N-[[4-chloro-8-[(4-methylpiperazin-1-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methyl]carbamate (1.1 g, 2.0 mmol, 1.0 equiv.) in dioxane was added HCl (10.0 mL, 19.1 equiv.; 4 M solution in dioxane), and the mixture was stirred at RT for 3 h. The reaction mixture was concentrated under reduced pressure to give the title compound (1.1 g, crude) as a white solid.

[0340]

[0383] LCMS[M+1, M+3]=427.4, 429.4.

[0384] 1 H NMR (400 MHz, methanol-d4) δ = 8.76 (s, 1H), 8.13–8.02 (m, 2H), 5.71 (q, J = 8.4 Hz, 2H), 4.75 (s, 2H), 4.55 (s, 2H), 3.87–3.67 (m, 8H), 3.02 (s, 3H).

[0385] Step 5. N-[[4-chloro-8-[(4-methylpiperazin-1-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methyl]-1-methyl-pyrazole-4-carboxamide:

[0386] A solution of 1-methylpyrazole-4-carboxylic acid (3.0 g, 23.7 mmol, 1.0 equiv) in SOCl (30.0 mL, 413 mmol, 17.3 equiv) was stirred at 80° C. for 30 min. The reaction mixture was concentrated under reduced pressure to give 1-methylpyrazole-4-carbonyl chloride (3.5 g, crude) as a white solid. This material was used in the next phase of this step without further purification.

[0341]

[0387] To a solution of [4-chloro-8-[(4-methylpiperazin-1-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methanamine hydrochloride (500 mg, 1.0 mmol, 1.0 equiv) in THF (0.5 mL) was added TEA (600 μL, 4.3 mmol, 4.0 equiv). The mixture was stirred at RT for 30 minutes, and then 1-methylpyrazole-4-carbonyl chloride (156.0 mg, 1.0 mmol, 1.0 equiv) was added. The mixture was stirred at RT for 10.5 hours. The reaction mixture was poured into HO (5 mL) and extracted with ethyl acetate (3 × 5 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Waters Xbridge BEH C18 100 mm × 30 mm × 10 μm; mobile phase: 20-35% acetonitrile in water (+10 mM NH4HCO3)) to give the title compound (300 mg, 50% yield) as a white solid.

[0342]

[0388] LCMS[M+1, M+3]=535.2, 537.2.

[0389] 1 H NMR (400 MHz, DMSO-d6) δ 8.81 (t, J = 5.6 Hz, 1H), 8.17 (s, 1H), 8.14 (s, 1H), 7.97 (d, J = 8.8 Hz, 1H), 7.91 (s, 1H), 7.82 - 7.66 (m, 1H), 5.70 (q, J = 8.8 Hz, 2H), 4.72 (d, J = 6.0 Hz, 2H), 3.87 (s, 3H), 3.64 (s, 2H), 2.47 - 2.19 (m, 8H), 2.15 (s, 3H).

[0343] Example 168 N-[[4-chloro-8-[[3-(dimethylamino)azetidin-1-yl]methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methyl]-1-methyl-pyrazole-4-carboxamide

[0344] [ka]

[0345]

[0390] Example 168 was prepared in a manner similar to that described for Example 167.

[0391] LCMS: [M+1, M+3]=535.2, 537.2.

[0346]

[0392] 1 H NMR (400 MHz, chloroform-d) δ 8.28 (s, 1H), 7.93 (s, 2H), 7.73 (dd, J = 1.2, 8.4 Hz, 1H), 7.52 (d, J = 8.4 Hz, 1H), 7.12 (s, 1H), 5.32 (q, J = 8.0 Hz, 2H), 4.99 (d, J = 4.8 Hz, 2H), 3.99 (s, 3H), 3.83 (s, 2H), 3.59 - 3.49 (m, 2H), 3.03 - 2.95 (m, 2H), 2.95 - 2.84 (m, 1H), 2.13 (s, 6H).

[0347] Example 169 2-[4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]-N-methyl-acetamide

[0348] [ka]

[0349]

[0393] Scheme 9, Step 1. Ethyl 3-[(5-bromo-2-carbamoyl-1H-indol-3-yl)amino]-3-oxo-propanoate:

[0394] 3-Amino-5-bromo-1H-indole-2-carboxamide (10.0 g, 39.3 mmol, 1.0 equiv.) and diethyl propanedioate (74.6 mL, 492 mmol, 12.5 equiv.) were combined and heated at 200° C. for 30 min under solvent-free conditions. EtOH (300 mL) was then added, refluxed, and continued for an additional 2 h. The mixture was cooled to RT, and the suspension was filtered through filter paper to give the title compound (12.5 g, 84% yield) as a brown solid.

[0350]

[0395] LCMS[M+1, M+3]=368.2, 370.2.

[0396] Scheme 9, Step 2. Ethyl 2-(8-bromo-4-chloro-5H-pyrimido[5,4-b]indol-2-yl)acetate:

[0397] To a solution of ethyl 3-[(5-bromo-2-carbamoyl-1H-indol-3-yl)amino]-3-oxo-propanoate (16.2 g, 44.0 mmol, 1.0 equiv.) in dioxane (160 mL) was added POCl3 (61.5 mL, 660.0 mmol, 15.0 equiv.). The mixture was stirred at 90 °C for 12 h. The reaction mixture was cooled to RT and concentrated under reduced pressure. The residue was then treated with water and the pH was adjusted to pH = 8. The suspension was filtered through filter paper to give the title compound (16.0 g, 89% yield) as a brown solid.

[0351]

[0398] LCMS[M+1, M+3]=368.1, 370.1.

[0399] Scheme 9, Step 3. Ethyl 2-[8-bromo-4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]acetate:

[0400] To a solution of ethyl 2-(8-bromo-4-chloro-5H-pyrimido[5,4-b]indol-2-yl)acetate (12.0 g, 32.5 mmol, 1.0 equiv.) and 2,2,2-trifluoroethyl trifluoromethanesulfonate (7.5 g, 32.5 mmol, 1.0 equiv.) in DMF (150 mL) was added K2CO3 (13.5 g, 97.6 mmol, 3.0 equiv.). The mixture was stirred at RT for 12 h. The reaction mixture was diluted with water (200 mL), and the suspension was filtered through filter paper. The filter cake was triturated with ethyl acetate for 30 min to give the title compound (12.0 g, 69% yield) as a brown solid.

[0352]

[0401] LCMS[M+1, M+3]=450.1, 452.1.

[0402] Scheme 9, Step 4. Methyl 2-[8-bromo-4-methoxy-5-(2,2,2-trifluoroethyl)-pyrimido[5,4-b]indol-2-yl]acetate:

[0403] To a solution of ethyl 2-[8-bromo-4-chloro-5-(2,2,2-trifluoroethyl)-pyrimido[5,4-b]indol-2-yl]acetate (14.0 g, 31.0 mmol, 1.0 equiv) in MeOH (140.0 mL) was added CHONa (5.5 g, 31.0 mmol, 1.0 equiv; 30% in MeOH). The mixture was heated to 60 °C for 30 minutes. The reaction mixture was diluted with water (500 mL) and extracted with ethyl acetate (3 × 500 mL × 3). The combined organic layers were washed with brine, dried over NaSO, filtered, and the filtrate was concentrated under reduced pressure to give the title compound (12.0 g, 65% yield) as a brown solid.

[0353]

[0404] LCMS[M+1, M+3]=432.1, 434.1.

[0405] Scheme 9, Step 5. Methyl 2-[8-[(dimethylamino)methyl]-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]acetate:

[0406] To a solution of methyl 2-[8-bromo-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]acetate (11.5 g, 26.6 mmol, 1.0 equiv.) and potassium ((dimethylamino)methyl)trifluoroborate (8.7 g, 53.2 mmol, 2.0 equiv.) in 2-methylbutan-2-ol (120 mL) and HO (30 mL) was added CsCO (17.3 g, 53.2 mmol, 2.0 equiv.). Then, [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (1.70 g, 2.60 mmol, 0.1 equiv.) was added, and the reaction mixture was purged with nitrogen. The mixture was stirred under a nitrogen atmosphere at 90 °C for 1 hour. The reaction mixture was diluted with water (200 mL) and extracted with EtOAc (3 × 200 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (5–100% ethyl acetate in petroleum ether) to afford the title compound (6.5 g, 52% yield) as a brown solid.

[0354]

[0407] LCMS[M+1]=411.1.

[0408] Scheme 9, Step 6. Methyl 2-[4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]acetate:

[0409] A mixture of methyl 2-[8-[(dimethylamino)methyl]-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]acetate (1.50 g, 3.60 mmol, 1.0 equiv) in POCl (60 mL) was degassed and purged with nitrogen, and the mixture was then stirred at 110 °C under a nitrogen atmosphere for 12 h. The reaction mixture was diluted with a saturated aqueous solution of NaHCO (200 mL) and extracted with EtOAc (3 × 200 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure to give the title compound (6.00 g, 81% yield) as a brown solid.

[0355]

[0410] LCMS[M+1, M+3]=415.3, 417.3.

[0411] Scheme 9, Step 7. 2-[4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]acetic acid:

[0412] To a solution of methyl 2-[4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]acetate (2.00 g, 4.80 mmol, 1.0 equiv.) in DCE (20 mL) was added hydroxy(trimethyl)stannane (5.20 g, 28.9 mmol, 6.0 equiv.). The mixture was stirred at 80 °C for 12 h. The reaction mixture was diluted with EtOAc (30 mL) and washed with HO (3 × 50 mL). The combined aqueous layers were lyophilized to give the residue. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 100 mm × 30 mm × 5 μm; mobile phase: 5–30% acetonitrile in HO (+0.2% formic acid)) to give the title compound (200 mg, 10% yield) as a white solid.

[0356]

[0413] LCMS[M+1, M+3]=401.2, 403.1.

[0414] 1 H NMR (400 MHz, DMSO-d6) δ 13.61 - 11.73 (m, 1H), 8.18 (s, 1H), 7.98 (d, J = 8.4 Hz, 1H), 7.77 (dd, J = 1.2, 8.8 Hz, 1H), 5.71 (q, J = 8.8 Hz, 2H), 4.02 (s, 2H), 3.62 (s, 2H), 2.20 (s, 6H).

[0415] Scheme 9, Step 8. 2-[4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]-N-methyl-acetamide:

[0416] To a solution of 2-[4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]acetic acid (100.0 mg, 249.5 μmol, 1.0 equiv.) in acetonitrile (0.5 mL) was added chloro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate (77.0 mg, 274.4 μmol, 1.1 equiv.), N-methylimidazole (61.6 μL, 773.4 μmol, 3.1 equiv.), and methanamine hydrochloride (20.2 mg, 299 μmol, 1.2 equiv.). The mixture was stirred at RT for 1 h. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (2 × 15 mL). The combined organic layers were washed with 15 mL of brine, dried over NaSO, filtered, and concentrated under reduced pressure to give a residue that was purified by preparative HPLC (column: Waters Xbridge BEH C18 100 mm × 30 mm × 10 μm; mobile phase: 15–45% acetonitrile in water (+10 mM NHHCO)) to give the title compound (15.8 mg, 15% yield) as a white solid.

[0357]

[0417] LCMS: [M+1, M+3]=413.9, 415.9.

[0418] 1H NMR (400 MHz, DMSO-d6) δ = 8.16 (s, 1H), 8.05 (br d, J = 3.6 Hz, 1H), 7.97 (d, J = 8.8 Hz, 1H), 7.76 (d, J = 8.4 Hz, 1H), 5.70 (q, J = 8.8 Hz, 2H), 3.88 (s, 2H), 3.59 (s, 2H), 2.63 (d, J = 4.8 Hz, 3H), 2.19 (s, 6H).

[0358] Example 170 2-[4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]-N-methyl-acetamide

[0359] [ka]

[0360]

[0419] Scheme 9, Step 8. 2-[4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]-N-(oxetan-3-yl)acetamide:

[0420] To a solution of 2-[4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]acetic acid (60.0 mg, 149.7 μmol, 1.0 equiv.) and oxetan-3-amine (13.1 mg, 179.6 μmol, 1.2 equiv.) in THF (1.0 mL) was added DIEA (78.2 μL, 449.1 μmol, 3.0 equiv.) and T4P (216 mg, 299 μmol, 2.0 equiv.; 50% solution in EtOAc). The mixture was stirred at RT for 2 h. The reaction mixture was poured into HO (3 mL) and extracted with EtOAc (3 × 5 mL). The combined organic layers were washed with HO (15 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Waters Xbridge C18 150 mm × 50 mm × 10 μm; mobile phase: 20-60% acetonitrile in H2O (+10 mM NH4HCO3)) to give the title compound (11.4 mg, 16% yield) as a white solid.

[0361]

[0421] LCMS: [M+1, M+3]=456.1, 458.1.

[0422] 1H NMR (400 MHz, DMSO-d6) δ 8.96 (d, J = 6.4 Hz, 1H), 8.16 (s, 1H), 7.98 (d, J = 8.8 Hz, 1H), 7.77 (d, J = 8.8 Hz, 1H), 5.71 (q, J = 8.8 Hz, 2H), 4.83 (q, J = 6.8 Hz, 1H), 4.77 - 4.70 (m, 2H), 4.48 (t, J = 6.4 Hz, 2H), 3.94 (s, 2H), 3.60 (s, 2H), 2.19 (s, 6H).

[0423]

[0362] [Table 8-1]

[0363] [Table 8-2]

[0364] [Table 8-3]

[0365] Example 181 2-[4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]-N-methyl-acetamide

[0366] [ka]

[0367]

[0424] Scheme 10, Step 1. Methyl 8-bromo-4-hydroxy-5H-pyrimido[5,4-b]indole-2-carboxylate:

[0425] To a solution of 3-amino-5-bromo-1H-indole-2-carboxamide (27 g, 106 mmol, 1 equiv.) in MeOH (270 mL) was added dimethyl oxalate (37.6 g, 319 mmol, 3 equiv.) and sodium methoxide (28.7 g, 159.4 mmol, 1.5 equiv.; 30% solution in MeOH). The reaction was stirred at 70° C. for 12 hours. Water (1,000 mL) was added to the reaction mixture, and the resulting precipitate was filtered and concentrated under reduced pressure to give the title compound (47.8 g, crude) as a brown solid.

[0368]

[0426] LCMS: [M+H, M+3]=322.1, 324.1.

[0427] Scheme 10, Step 2. Methyl 8-bromo-4-methoxy-5H-pyrimido[5,4-b]indole-2-carboxylate:

[0428] Methyl 8-bromo-4-hydroxy-5H-pyrimido[5,4-b]indole-2-carboxylate (20 g, 62.0 mmol, 1 equiv.) was added to POCl (200 mL) at 20° C. The solution was then stirred at 110° C. for 16 h. The reaction mixture was concentrated under reduced pressure to give the title compound (33 g, crude) as a brown solid.

[0369]

[0429] A solution of methyl 8-bromo-4-chloro-5H-pyrimido[5,4-b]indole-2-carboxylate (33 g, 96.9 mmol, 1 equiv.) in MeOH (350 mL) was then treated with NaOMe (26.1 g, 145.3 mmol, 1.5 equiv.; 30% solution in MeOH), and the reaction mixture was stirred at 70° C. for 2 h. The reaction was cooled to RT, diluted with EtOAc (500 mL), and then quenched with water. The layers were separated, and the aqueous layer was extracted with EtOAc (3×100 ml). The combined organic layers were dried over NaSO, filtered, and concentrated under reduced pressure. The residue was triturated with ACN (1000 mL), then filtered and dried to give the title compound (5.8 g, 18% yield) as a brown solid.

[0370]

[0430] LCMS: [M+H, M+3]=336.0, 338.1.

[0431] Scheme 10, Step 3. Methyl 8-bromo-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-2-carboxylate:

[0432] To a solution of methyl 8-bromo-4-methoxy-5H-pyrimido[5,4-b]indole-2-carboxylate (5.8 g, 17.2 mmol, 1 equiv.) and KCO (4.7 g, 34.5 mmol, 2 equiv.) in DMF (58 mL) was added 2,2,2-trifluoroethyl trifluoromethanesulfonate (4.81 g, 20.7 mmol, 1.2 equiv.). The reaction mixture was warmed to RT and stirred for 2 h. The reaction mixture was cooled to 0 °C and then quenched with HO (20 mL). The mixture was extracted with EtOAc (3 × 20 mL), and the combined organic layers were concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0–25% ethyl acetate in petroleum ether) to afford the title compound (2.3 g, 32% yield) as a brown solid.

[0371]

[0433] LCMS: [M+H, M+3]=418.0, 420.0.

[0434] Scheme 10, Step 4. 8-[(Dimethylamino)methyl]-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-2-carboxylic acid:

[0435] To a solution of methyl 8-bromo-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-2-carboxylate (2.3 g, 5.5 mmol, 1 equiv.) and potassium ((dimethylamino)methyl)trifluoroborate (3.6 g, 22.0 mmol, 4 equiv.) in 2-methylbutan-2-ol (23 mL) was added a solution of CsCO (3.5 g, 11.0 mmol, 2 equiv.) in HO (4.6 mL) and [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (358.4 mg, 550.0 μmol, 0.1 equiv.) under a nitrogen atmosphere at RT. The mixture was stirred at 90 °C for 1 h. The reaction mixture was diluted with water and extracted with EtOAc. The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The product was purified by preparative HPLC (Column: Waters Xbridge BEH C18 250 mm × 70 mm × 10 μm; Mobile phase: 1-40% acetonitrile in water (+10 mM NHHCO)) to give the title compound (2.0 g, 95% yield) as a white solid.

[0372]

[0436] LCMS: [M+H]=383.2.

[0437] Scheme 10, Step 5. Methyl 8-[(dimethylamino)methyl]-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-2-carboxylate:

[0438] 8-[(Dimethylamino)methyl]-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-2-carboxylic acid (1.5 g, 3.9 mmol, 1 equiv.) was dissolved in oxalyl chloride (15.0 mL, 171.3 mmol, 43.6 equiv.) and the reaction was stirred at RT for 1 h. The reaction mixture was concentrated and methanol was added. Removal of the solvent under reduced pressure afforded the title compound (1.1 g, 71% yield) as a yellow solid.

[0373]

[0439] LCMS: [M+H]=397.5.

[0440] Scheme 10, Step 6. Methyl 8-[(dimethylamino)methyl]-4-hydroxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-2-carboxylate:

[0441] To a solution of methyl 8-[(dimethylamino)methyl]-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-2-carboxylate (1 g, 2.5 mmol, 1 equiv.) in ACN (10 mL) was added TMSCl (480 μL, 3.7 mmol, 1.5 equiv.) and NaI (567 mg, 3.7 mmol, 1.5 equiv.) at RT. The reaction was stirred at RT for 1 h. Water (14 μL, 757 μmol, 0.3 equiv.) was then added, and the reaction mixture was stirred at 65° C. for 3 h. The reaction mixture was diluted with water (10 mL) and extracted with EtOAc (3×10 mL). The combined organic layers were dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (5:1 DCM:MeOH) to afford the title compound (800 mg, 83% yield) as a white solid.

[0374]

[0442] LCMS: [M+H]=383.1.

[0443] Scheme 10, Step 7. Methyl 4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-2-carboxylate:

[0444] Methyl 8-[(dimethylamino)methyl]-4-hydroxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-2-carboxylate (1 g, 2.6 mmol, 1 equiv.) was dissolved in POCl (20 mL, 214.5 mmol, 82.0 equiv.) and the mixture was stirred at 110 °C for 2 h. The solvent was removed under reduced pressure. The flask was cooled to 0 °C and a saturated aqueous solution of NaHCO was added to adjust the pH to pH = 8. The mixture was extracted with EtOAc (3 × 20 mL) and the combined organic layers were concentrated under reduced pressure. The reaction was purified by preparative TLC (100% THF) to afford the title compound (650 mg, 62% yield) as a yellow solid.

[0375]

[0445] LCMS: [M+H]=401.3.

[0446] Scheme 10, Step 8. 4-Chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-2-carboxylic acid:

[0447] To a solution of methyl 4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-2-carboxylate (100 mg, 250 μmol, 1 equiv.) in DCE (2 mL) was added hydroxy(trimethyl)stannane (271 mg, 1.5 mmol, 6 equiv.). The mixture was stirred at 80°C for 2 h. The mixture was concentrated under reduced pressure to give a residue, which was purified by preparative HPLC (column: Waters Xbridge BEH C18 100 mm × 30 mm × 10 μm; mobile phase: 1-40% acetonitrile in water (+10 mM NH4HCO3)) to give the title compound (20.6 mg, 20% yield) as a white solid.

[0376]

[0448] LCMS: [M+H]=387.1.

[0449] 1 H NMR (400 MHz, DMSO-d6) δ = 8.34 (s, 1H), 8.03 (d, J = 8.8 Hz, 1H), 7.82 (dd, J = 1.2 8.8 Hz 1H), 5.75 (q, J = 8.7 Hz, 2H), 3.87 (s, 2H), 2.38 (s, 6H).

[0450] Scheme 10, Step 9. 4-Chloro-8-[(dimethylamino)methyl]-N,N-dimethyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-2-carboxamide:

[0451] To a solution of 4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-2-carboxylic acid (30 mg, 70.9 μmol, 1.0 equiv., HCl) and dimethylamine (28.35 μL, 0.8 equiv.; 2 M solution in THF) in DMF (1 mL) was added DIEA (37 μL, 213 μmol, 3.0 equiv.) and HATU (54 mg, 142 μmol, 2.0 equiv.). The mixture was stirred at RT for 2 h. The reaction was quenched with (2 mL). The aqueous layer was extracted with ethyl acetate (3 × 1 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex luna C18 80 mm × 30 mm × 3 μm; mobile phase: 5-35% acetonitrile in water (+0.04% HCl)) to give the title compound (9 mg, 29% yield) as a white solid.

[0377]

[0452] LCMS[M+1, M+3]=414.3, 416.3.

[0453] 1 H NMR (400 MHz, DMSO-d6) δ = 8.58 (s, 1H), 8.18 (d, J = 8.8 Hz, 1H), 8.05 (d, J = 8.4 Hz, 1H), 5.81 (q, J = 8.8 Hz, 2H), 4.49 (s, 2H), 3.07 (s, 3H), 2.89 (s, 3H), 2.74 (s, 6H).

[0454]

[0378] [Table 9]

[0379] Example 185 4-chloro-8-(2,5-diazaspiro[3.4]octan-2-ylmethyl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole

[0380] [ka]

[0381]

[0455] Scheme 11, Step 1. tert-Butyl 2-[[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]methyl]-2,5-diazaspiro[3.4]octane-5-carboxylate:

[0456] To a solution of 4-chloro-8-(chloromethyl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole (200 mg, 599 μmol, 1 equiv.) in acetonitrile (4.0 mL) was added TEA (500 μL, 3.6 mmol, 6.0 equiv.) and tert-butyl 2,5-diazaspiro[3.4]octane-5-carboxylate-oxalate (154.0 mg, 299.3 μmol, 0.5 equiv.). The mixture was stirred at RT for 7 h. The reaction mixture was diluted with HO (10 mL) and extracted with EtOAc (3 × 5 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO, 10:1 DCM:MeOH) to afford the title compound (46 mg, 13% yield) as a white solid.

[0382]

[0457] LCMS[M+1, M+3]=510.2, 512.2.

[0458] Scheme 11, Step 2. 4-Chloro-8-(2,5-diazaspiro[3.4]octan-2-ylmethyl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole:

[0459] A solution of tert-butyl 2-[[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]methyl]-2,5-diazaspiro[3.4]octane-5-carboxylate (130 mg, 255 μmol, 1.0 equiv.) in TFA (0.4 mL) and DCM (2.0 mL) was stirred at RT for 1 h. The reaction mixture was diluted with saturated aqueous Na2CO3 (15 mL) and extracted with EtOAc (8 mL). The layers were separated, and the organic extract was filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Waters Xbridge C18 150 mm × 50 mm × 10 μm; mobile phase: 25–55% acetonitrile in water (+10 mM NH4HCO3)) to afford the title compound (85 mg, 80% yield) as a white solid.

[0383]

[0460] LCMS[M+1, M+3]=410.1, 412.1.

[0461] 1 H NMR (400 MHz, chloroform-d) δ 8.93 (s, 1H), 8.30 (s, 1H), 7.75 (d, J = 8.8 Hz, 1H), 7.53 (d, J = 8.4 Hz, 1H), 5.34 (q, J = 8.0 Hz, 2H), 3.82 (s, 2H), 3.41 (d, J = 8.0 Hz, 2H), 3.09 (d, J = 8.0 Hz, 2H), 2.93 (t, J = 6.8 Hz, 2H), 2.01 (t, J = 6.8Hz, 2H), 1.78 - 1.73 (m, 2H). Example 186 4-chloro-8-(piperazin-1-ylmethyl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole

[0384] [ka]

[0385]

[0462] Scheme 12, Step 1. tert-Butyl 4-[[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]methyl]piperazine-1-carboxylate:

[0463] To a mixture of 4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-8-carbaldehyde (55.0 mg, 175 μmol, 1.0 equiv.) and tert-butyl piperazine-1-carboxylate (65.3 mg, 351 μmol, 2.0 equiv.) in DCE (1 mL) was added AcOH (10.0 μL, 175.3 μmol, 1.0 equiv.), and the mixture was stirred for 15 min. NaBH(OAc) (37.1 mg, 175.3 μmol, 1.0 equiv.) was then added, and the reaction was stirred at RT for 1 h. The reaction was quenched by the addition of HO (10 mL), and the mixture was then extracted with DCM (3 × 5 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO 2 , 1:1 ethyl acetate:petroleum ether) to afford the title compound (38.0 mg, 45% yield) as a white solid.

[0386]

[0464] LCMS: [M+1, M+3]=484.4, 486.4.

[0465] Scheme 12, Step 2. 4-Chloro-8-(piperazin-1-ylmethyl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole:

[0466] A solution of HCl in dioxane (0.2 mL, 4 M) was added to a flask containing tert-butyl 4-[[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]methyl]piperazine-1-carboxylate (35.0 mg, 72.3 μmol, 1.0 equiv.), and the reaction was stirred at RT for 30 min. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 75 mm × 30 mm × 3 μm; mobile phase: 5–35% acetonitrile in water (+0.04% HCl)) to afford the title compound (10.0 mg, 36% yield) as a white solid (hydrochloride salt).

[0387]

[0467] LCMS: [M+1, M+3]=384.1, 386.1.

[0468] 1 H NMR (400 MHz, DMSO-d6) δ 12.49 - 11.69 (br s, 1H), 9.57 (br s, 2H), 8.99 (s, 1H), 8.65 (s, 1H), 8.14 (q, J = 3.6 Hz, 2H), 5.79 (q, J = 8.8 Hz, 2H), 4.63 (br s, 2H), 3.72 - 3.12 (m, 8H).

[0469]

[0388] [Table 10-1]

[0389] [Table 10-2]

[0390] [Table 10-3]

[0391] [Table 10-4]

[0392] Example 199 4-chloro-8-[(5-methyl-2,5-diazaspiro[3.4]octan-2-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole

[0393] [ka]

[0394]

[0470] 4-chloro-8-[(5-methyl-2,5-diazaspiro[3.4]octan-2-yl)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole:

[0471] To a solution of 4-chloro-8-(2,5-diazaspiro[3.4]octan-2-ylmethyl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole (35.0 mg, 85.4 μmol, 1.0 equiv.) in DCE (1.0 mL) was added formaldehyde (3.2 μL, 42.7 μmol, 0.5 equiv.; 37% solution in water) and HOAc (4.9 μL, 85.4 μmol, 1.0 equiv.). The mixture was stirred at RT for 1 h. An additional portion of NaBH(OAc) (36.2 mg, 170.8 μmol, 2.0 equiv.) was added, and the mixture was stirred at RT for 2 h. The reaction mixture was diluted with HO (5 mL) and extracted with EtOAc (3 × 3 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Waters Xbridge BEH C18 100 mm × 30 mm × 10 μm; mobile phase: 30-60% acetonitrile in water (+10 mM NH4HCO3)) to give the title compound (8.0 mg, 22% yield) as a white solid.

[0395]

[0472] LCMS[M+1, M+3]=424.1, 426.1.

[0473] 1H NMR (400 MHz, chloroform-d) δ 8.93 (s, 1H), 8.31 (s, 1H), 7.75 (s, 1H), 7.55 (s, 1H), 5.34 (br d, J = 8.0 Hz, 2H), 3.83 (s, 2H), 3.34 (s, 2H), 3.16 (s, 2H), 2.77 - 2.61 (m, 2H), 2.48 (s, 3H), 2.22 - 2.05 (m, 2H), 1.77 - 1.73 (m, 2H).

[0474]

[0396] [Table 11]

[0397] Example 203 1-[4-chloro-7-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine

[0398] [ka]

[0399]

[0475] Scheme 13, Step 1. 5-Bromo-2-[(4-methoxyphenyl)methylamino]-4-methyl-benzonitrile:

[0476] To a mixture of 5-bromo-2-fluoro-4-methyl-benzonitrile (15.0 g, 70.0 mmol, 1.0 equiv) and para-methoxybenzylamine (18.1 mL, 140.1 mmol, 2.0 equiv) in DMSO (150 mL) was added K2CO3 (29.0 g, 210.2 mmol, 3.0 equiv). The mixture was stirred at 80 °C for 2 h. The reaction mixture was cooled to RT and quenched with water (300 mL). The mixture was extracted with EtOAc (3 × 150 mL). The combined organic layers were washed with brine (3 × 150 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the title compound (12.2 g, 53% yield) as a white solid.

[0400]

[0477] 1 H NMR (400 MHz, DMSO-d6) δ 7.74 (s, 1H), 7.35 (d, J = 8.5 Hz, 2H), 6.96 (d, J = 8.6 Hz, 2H), 6.75 (s, 1H), 4.41 (d, J = 6.0 Hz, 2H), 3.79 (s, 3H), 2.60 - 2.55 (m, 2H), 2.28 (s, 3H).

[0478] Scheme 13, Step 2. 2-Amino-5-bromo-4-methyl-benzonitrile:

[0479] A mixture of 5-bromo-2-[(4-methoxyphenyl)methylamino]-4-methyl-benzonitrile (12.0 g, 36.2 mmol, 1.0 equiv) in DCM (60 mL) and TFA (30 mL) was stirred at RT for 30 min. The reaction mixture was concentrated and then quenched by the addition of a saturated aqueous solution of NaHCO3 (90 mL) at RT. The mixture was extracted with DCM. The extract was dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography (0-10% ethyl acetate in petroleum ether) to afford the title compound (7.0 g, 90% yield) as a yellow solid.

[0401]

[0480] 1 H NMR (400 MHz, DMSO-d6) δ 7.58 (s, 1H), 6.74 (s, 1H), 6.14 (br s, 2H), 2.23 (s, 3H).

[0481] Scheme 13, Step 3. Ethyl 2-(4-bromo-2-cyano-5-methyl-anilino)acetate:

[0482] To a mixture of 2-amino-5-bromo-4-methyl-benzonitrile (4.0 g, 18.9 mmol, 1.0 equiv) and ethyl 2-oxoacetate (11.6 g, 56.8 mmol, 3.0 equiv) in DMF (50 mL) was added TMSCl (12.3 g, 113.7 mmol, 14.43 mL, 6.0 equiv), and the mixture was stirred at RT for 2 h. NaBH(OAc) (10.0 g, 47.3 mmol, 2.5 equiv) was then added, and the mixture was stirred at RT for 12 h. The reaction mixture was quenched by the addition of a saturated aqueous solution of NaHCO (150 mL) at RT and then extracted with DCM (3 × 60 mL). The combined organic layers were washed with brine (3 × 100 mL), dried over NaSO, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex luna C18 250 mm × 150 mm × 15 μm; mobile phase: 50-85% acetonitrile in water (+0.05% HCl)) to give the title compound (2.2 g, 39% yield) as a yellow solid.

[0402]

[0483] LCMS: [M+1, M+3]=296.9, 298.8.

[0484] 1 H NMR (400 MHz, DMSO-d6) δ 7.72 (s, 1H), 6.73 (s, 1H), 6.43 (t, J = 6.0 Hz, 1H), 4.18 - 4.10 (m, 2H), 4.05 (d, J = 6.0 Hz, 2H), 2.29 (s, 3H), 1.24 - 1.17 (m, 3H).

[0485] Scheme 13, Step 4. 1-(tert-butyl) 2-ethyl 3-amino-5-bromo-6-methyl-1H-indole-1,2-dicarboxylate:

[0486] To a mixture of ethyl 2-(4-bromo-2-cyano-5-methyl-anilino)acetate (2.2 g, 7.4 mmol, 1.0 equiv) in DCM (20 mL) was added TEA (5.1 mL, 37.0 mmol, 5.0 equiv), BocO (9.7 g, 44.4 mmol, 6.0 equiv), and DMAP (90 mg, 740 μmol, 0.1 equiv). The mixture was stirred at RT for 12 h. The reaction mixture was quenched by the addition of a saturated aqueous solution of NaHCO (60 mL). The mixture was extracted with EtOAc (3 × 30 mL), and the combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0–7% ethyl acetate in petroleum ether) to afford the title compound (2.6 g, 83% yield) as a yellow solid.

[0403]

[0487] LCMS: [M-55, M-53]=397.2, 399.2.

[0488] 1 H NMR (400 MHz, DMSO-d6) δ 8.16 (s, 1H), 7.50 (d, J = 15.6 Hz, 1H), 4.38 (d, J = 16.4 Hz, 2H), 4.23 - 4.03 (m, 2H), 2.40 (s, 3H), 1.43 - 1.40 (m, 3H), 1.38 (s, 9H).

[0489] Scheme 13, Step 5. 1-(tert-butyl) 2-ethyl-5-bromo-3-(((dimethylamino)methylene)amino)-6-methyl-1H-indole-1,2-dicarboxylate:

[0490] To a mixture of 1-(tert-butyl) 2-ethyl 3-amino-5-bromo-6-methyl-1H-indole-1,2-dicarboxylate (2.6 g, 6.5 mmol, 1.0 equiv.) in DMF (25 mL) was added 1,1-dimethoxy-N,N-dimethyl-methanamine (1.1 mL, 8.5 mmol, 1.3 equiv.). The mixture was stirred at 100° C. for 4 h. The reaction mixture was cooled to RT, diluted with water, and then extracted with EtOAc (3×100 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure to give the title compound (2.9 g, 98% yield) as a yellow oil.

[0404]

[0491] LCMS: [M+1, M+3]=452.2, 454.2.

[0492] Scheme 13, Step 6. 8-Bromo-7-methyl-5H-pyrimido[5,4-b]indol-4-ol:

[0493] A mixture of 1-(tert-butyl) 2-ethyl-5-bromo-3-(((dimethylamino)methylene)amino)-6-methyl-1H-indole-1,2-dicarboxylate (2.9 g, 6.4 mmol, 1.0 equiv) in MeOH (10 mL) and NH HO (20 mL) was stirred at 70 °C for 48 h. The mixture was cooled to RT, and the solid was filtered and dried under reduced pressure to give the title compound (900 mg, 48% yield) as a gray solid.

[0405]

[0494] LCMS: [M+1, M+3]=278.0, 279.9.

[0495] Scheme 13, Step 7. 8-Bromo-4-chloro-7-methyl-5H-pyrimido[5,4-b]indole:

[0496] A mixture of 8-bromo-7-methyl-5H-pyrimido[5,4-b]indol-4-ol (900.0 mg, 3.2 mmol, 1.0 equiv) in POCl (9 mL) was stirred for 12 h at 120° C. The reaction mixture was concentrated under reduced pressure to give the title compound (700 mg, 73% yield) as a yellow solid.

[0406]

[0497] LCMS: [M+1, M+3]=296.1, 298.2.

[0498] 1 H NMR (400 MHz, DMSO-d6) δ 12.47 (s, 1H), 8.85 (s, 1H), 8.41 (s, 1H), 7.67 (s, 1H), 2.57 (s, 3H).

[0499] Scheme 13, Step 8. 8-Bromo-4-chloro-7-methyl-5-(2,2,2-trifluoroethyl)-pyrimido[5,4-b]indole:

[0500] To a solution of 8-bromo-4-chloro-7-methyl-5H-pyrimido[5,4-b]indole (200 mg, 674 μmol, 1.0 equiv.) in DMF (2 mL) was added KCO (373 mg, 2.7 mmol, 4.0 equiv.) and 2,2,2-trifluoroethyl trifluoromethanesulfonate (313 mg, 1.3 mmol, 2.0 equiv.), and the mixture was stirred at RT for 20 h. The reaction mixture was quenched with a saturated aqueous solution of NHCl (10 mL), and then the mixture was extracted with EtOAc (3 × 8 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO, 1:1 ethyl acetate:petroleum ether) to afford the title compound (100.0 mg, 38% yield) as a yellow solid.

[0407]

[0501] LCMS: [M+1, M+3]=378.1, 380.1.

[0502] Scheme 13, Step 9. 1-[4-chloro-7-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine:

[0503] To a mixture of 8-bromo-4-chloro-7-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole (100 mg, 264 μmol, 1.0 equiv.) and potassium ((dimethylamino)methyl)trifluoroborate (174 mg, 1.0 mmol, 4.0 equiv.) in 2-methylbutan-2-ol (0.8 mL), a solution of CsCO (172 mg, 528 μmol, 2.0 equiv.) in HO (0.2 mL) and [1,1′-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (17.2 mg, 26.4 μmol, 0.1 equiv.) was added under a nitrogen atmosphere. The mixture was stirred at 80° C. for 1 h. The reaction mixture was cooled to RT, diluted with HO (10 mL), and then extracted with EtOAc (3×8 mL). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex luna C18 80 mm × 30 mm × 3 μm; mobile phase: 10–40% acetonitrile in water (+0.04% HCl)) to give the title compound (11.1 mg, 11% yield; hydrochloride salt) as a yellow solid.

[0408]

[0504] LCMS: [M+1, M+3]=357.1, 359.1.

[0505] 1 H NMR (400 MHz, DMSO-d6) δ 10.04 (br s, 1H), 8.96 (s, 1H), 8.55 (s, 1H), 8.02 (s, 1H), 5.73 (q, J = 8.8 Hz, 2H), 4.53 (d, J = 5.6 Hz, 2H), 2.81 (d, J = 4.8 Hz, 6H), 2.69 (s, 3H).

[0409] Example 204 2-[4-[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]pyrazol-1-yl]-N,N-dimethyl-ethanamine

[0410] [ka]

[0411]

[0506] Scheme 14, Step 1. 2-[4-[4-Methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]pyrazol-1-yl]-N,N-dimethyl-ethanamine:

[0507] To a solution of 8-iodo-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole (180 mg, 442 μmol, 1.0 equiv.) and N,N-dimethyl-2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl]ethanamine (176 mg, 663 μmol, 1.5 equiv.) in DMF (2.0 mL) and HO (0.4 mL) was added KCO (183.3 mg, 1.3 mmol, 3.0 equiv.) and Pd(dppf)Cl·CHCl (36.1 mg, 44.2 μmol, 0.1 equiv.) under a nitrogen atmosphere at RT. The mixture was stirred at 110 °C for 1 h. The mixture was cooled to RT and poured into HO (5 mL). The aqueous phase was extracted with ethyl acetate (3 × 5 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO, 10:1 DCM:methanol) to afford the title compound (170 mg, 90% yield) as a black solid.

[0412]

[0508] LCMS[M+1]=419.1.

[0509] 1H NMR (400 MHz, methanol-d4) δ 8.66 (s, 1H), 8.41 (s, 1H), 8.16 (s, 1H), 7.96 (s, 1H), 7.92 (dd, J = 1.2, 8.8 Hz, 1H), 7.75 (d, J = 8.4 Hz, 1H), 5.39 (q, J = 8.8 Hz, 2H), 4.36 (t, J = 6.4 Hz, 2H), 4.26 (s, 3H), 2.90 (t, J = 6.8 Hz, 2H), 2.34 (s, 6H).

[0510] Scheme 14, Step 2. 2-[4-[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]pyrazol-1-yl]-N,N-dimethyl-ethanamine:

[0511] 2-[4-[4-Methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]pyrazol-1-yl]-N,N-dimethyl-ethanamine (0.1 g, 239 μmol, 1.0 equiv.) and dichlorophosphorylbenzene (1.0 mL) were combined at RT, and the mixture was stirred at 140 °C for 1 h. The reaction mixture was filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150 mm × 25 mm × 10 μm; mobile phase: 15–45% acetonitrile in water (+0.2% formic acid)) to afford the title compound (33.8 mg, 33% yield) as a white solid.

[0413]

[0512] LCMS[M+1, M+3]=423.3, 425.2.

[0513] 1 H NMR (400 MHz, DMSO-d6) δ 8.94 (s, 1H), 8.48 (s, 1H), 8.37 (s, 1H), 8.10 - 8.03 (m, 3H), 5.75 (q, J = 8.8 Hz, 2H), 4.23 (t, J = 6.4 Hz, 2H), 2.71 (t, J = 6.4 Hz, 2H), 2.20 (s, 6H).

[0414] Example 205 4-chloro-8-(1,2,3,4-tetrahydroisoquinolin-7-yl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole

[0415] [ka]

[0416]

[0514] Scheme 14, Step 1. tert-Butyl 7-[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-3,4-dihydro-1H-isoquinoline-2-carboxylate:

[0515] To a solution of 4-chloro-8-iodo-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole (50.0 mg, 121.4 μmol, 1.0 equiv.) and tert-butyl 7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydro-1H-isoquinoline-2-carboxylate (39.2 mg, 109.3 μmol, 0.9 equiv.) in DMF (0.5 mL) and HO (0.1 mL) was added Pd(PPh)Cl (8.5 mg, 12.1 μmol, 0.1 equiv.) and KCO (50 mg, 364 μmol, 3.0 equiv.). The mixture was stirred at 50° C. for 30 min. The mixture was cooled to RT and poured into HO (5 mL). The aqueous phase was extracted with ethyl acetate (3 × 5 mL). The combined organic layers were washed with brine, dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO, 3:1 petroleum ether:ethyl acetate) to give the title compound (50.0 mg, 80% yield) as a white solid.

[0417]

[0516] LCMS[M+1, M+3]=517.3, 519.4.

[0517] 4-chloro-8-(1,2,3,4-tetrahydroisoquinolin-7-yl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole:

[0518] tert-Butyl 7-[4-chloro-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-3,4-dihydro-1H-isoquinoline-2-carboxylate (45.0 mg, 87.0 μmol, 1.0 equiv.) was treated with HCl (2.0 mL; 4 M solution in dioxane), and the mixture was stirred at RT for 1 h. The reaction mixture was concentrated under reduced pressure. The crude product was triturated with acetonitrile, the mixture was filtered, and the filter cake was concentrated under reduced pressure to afford the title compound (30.0 mg, 83% yield; hydrochloride salt) as a white solid.

[0418]

[0519] LCMS[M+1, M+3]=417.0, 419.0.

[0520] 1 H NMR (400 MHz, DMSO-d6) δ = 9.43 (br s, 2H), 8.98 (s, 1H), 8.57 (s, 1H), 8.17 (s, 2H), 7.79 - 7.73 (m, 2H), 7.37 (br d, J = 8.4 Hz, 1H), 5.86 - 5.73 (m, 2H), 4.36 (br s, 2H), 3.42 (br d, J = 3.6 Hz, 2H), 3.07 (br t, J = 6.0 Hz, 2H).

[0419] Example 206 4-chloro-8-(6-methylpyridazin-3-yl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole

[0420] [ka]

[0421]

[0521] Scheme 14, Step 1. 4-Chloro-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole:

[0522] To a solution of 4-chloro-8-iodo-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole (200 mg, 486 μmol, 1.0 equiv.) and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (148 mg, 583 μmol, 1.2 equiv.) in dioxane (2.0 mL) was added KOAc (143 mg, 1.4 mmol, 3.0 equiv.) and Pd(dppf)Cl2·CHCl2 (39.6 mg, 48.6 μmol, 0.1 equiv.). The mixture was stirred at 100 °C under a nitrogen atmosphere for 16 h. The mixture was cooled to RT and poured into HO (4 mL). The aqueous phase was extracted with EtOAc (3 × 4 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The crude product was purified by preparative TLC (SiO, 2:1 petroleum ether:ethyl acetate) to give the title compound (80.0 mg, 40% yield) as a yellow solid.

[0422]

[0523] LCMS[M+1, M+3]=412.1, 414.1.

[0524] 1 H NMR (400 MHz, chloroform-d) δ 8.95 (s, 2H), 8.19 (dd, J = 0.8, 8.8 Hz, 1H), 7.57 (d, J = 8.0Hz, 1H), 5.37 (q, J = 8.0 Hz, 2H), 1.28 - 1.25 (m, 12H).

[0525] Scheme 14, Step 2. 4-Chloro-8-(6-methylpyridazin-3-yl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole:

[0526] To a mixture of 3-bromo-6-methylpyridazine (42 mg, 243 μmol, 2.0 equiv.), K2CO3 (50 mg, 364 μmol, 3.0 equiv.), 4-chloro-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole (50.0 mg, 121.4 μmol, 1.0 equiv.) in dioxane (0.5 mL) and HO (0.1 mL) was added Pd(dppf)Cl2·CHCl2 (9.9 mg, 12.1 μmol, 0.1 equiv.). The mixture was degassed with nitrogen and then stirred at 90 °C under a nitrogen atmosphere for 4 h. The reaction mixture was cooled to RT and filtered. The residue was triturated with acetonitrile and filtered to give the title compound (20.0 mg, 44% yield) as a gray solid.

[0423]

[0527] LCMS[M+1, M+3]=378.1, 380.1.

[0528] 1 H NMR (400 MHz, DMSO-d6) δ 9.07 (d, J = 1.2 Hz, 1H), 9.01 (s, 1H), 8.66 (dd, J = 1.6, 8.8 Hz, 1H), 8.37 (d, J = 8.8 Hz, 1H), 8.23 ​​(d, J = 8.8 Hz, 1H), 7.70 (d, J = 9.2 Hz, 1H), 5.82 (q, J = 8.8 Hz, 2H), 2.69 (s, 3H).

[0529]

[0424] [Table 12-1]

[0425] [Table 12-2]

[0426] [Table 12-3]

[0427] [Table 12-4]

[0428] Example 221 N-[[4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methyl]-2-methoxy-4-methylsulfonyl-aniline

[0429] [ka]

[0430]

[0530] Step 1. 1-[4-Methoxy 5-(2,2,2-trifluoroethyl)-2-vinyl-pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine:

[0531] A mixture of 1-[2-chloro-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine (2.0 g, 5.3 mmol, 1.0 equiv.), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (3.3 g, 21.4 mmol, 4 equiv.), KCO (742 mg, 5.3 mmol, 1.0 equiv.) in dioxane (10.0 mL) and HO (1.0 mL) was degassed with nitrogen. Pd(PPh) (620 mg, 537 μmol, 0.1 equiv.) was then added, and the mixture was again degassed with nitrogen. The mixture was stirred at 100 °C under a nitrogen atmosphere for 2 h. The reaction mixture was cooled to RT and quenched with water. The mixture was diluted with EtOAc and extracted with EtOAc (3 × 10 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (1–30% ethyl acetate in petroleum ether) to afford the title compound (1.0 g, 36% yield) as a yellow solid.

[0431]

[0532] LCMS[M+1]=365.4.

[0533] Step 2. 1-[4-chloro-5-(2,2,2-trifluoroethyl)-2-vinyl-pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine:

[0534] A solution of 1-[4-methoxy-5-(2,2,2-trifluoroethyl)-2-vinyl-pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine (200 mg, 549 μmol, 1 equiv.) in dichlorophosphorylbenzene (2 mL) was stirred at 120° C. for 1 h. The reaction mixture was cooled to RT and quenched with a saturated aqueous solution of NaHCO. The mixture was then diluted with HO (30 mL) and extracted with EtOAc (3×30 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The crude title compound (200 mg, 69% yield) was obtained as a yellow oil, which was used in the next step without further purification.

[0432]

[0535] LCMS[M+1, M+3]=369.5, 371.5.

[0536] Step 3. 4-Chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-2-carbaldehyde:

[0537] To a solution of 1-[4-chloro-5-(2,2,2-trifluoroethyl)-2-vinyl-pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine (450 mg, 1.2 mmol, 1.0 equiv.) in dioxane (5.0 mL) and HO (2.5 mL), KOsO·HO (22.5 mg, 61 μmol, 0.05 equiv.), NaIO (3.1 g, 14.6 mmol, 12 equiv.), and 2,6-dimethylpyridine (1.14 mL, 9.7 mmol, 8 equiv.) were added. The mixture was stirred at 20 °C for 1 h. The reaction was diluted with ethyl acetate, filtered, and concentrated under reduced pressure to afford the title compound (450.0 mg, 99% yield) as a yellow solid, which was used in the next step without further purification.

[0433]

[0538] LCMS[M+1, M+3]=371.3, 373.3.

[0539] Step 4. N-[[4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methyl]-2-methoxy 4-methylsulfonyl-aniline:

[0540] To a solution of 4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole-2-carbaldehyde (200.0 mg, 539.4 μmol, 1 equiv.) and 2-methoxy-4-methylsulfonyl-aniline (76 mg, 378 μmol, 0.7 equiv.) in DCE (0.5 mL) was added AcOH (62 μL, 1.0 mmol, 2 equiv.) and NaBH(OAc) (286 mg, 1.35 mmol, 2.5 equiv.). The mixture was stirred at RT for 2 h. The mixture was diluted with water, and the mixture was extracted with ethyl acetate. The extract was dried over NaSO, filtered, and concentrated. The residue was purified by preparative HPLC (Column 1: Phenomenex Luna C18 80 mm × 30 mm × 3 μm; Mobile phase: 15-45% acetonitrile in water (+0.04% HCl)), followed by a second purification by Column 2: Waters Xbridge BEH C18 100 mm × 30 mm × 10 μm; Mobile phase: 30-60% acetonitrile in water (+10 mM NH4HCO3)) to afford the title compound (10.4 mg, 50% yield) as a white solid.

[0434]

[0541] LCMS[M+1, M+3]=556.0, 558.0.

[0542] 1H NMR (400 MHz, DMSO-d6) δ = 8.16 (s, 1H), 7.98 (d, J = 8.8 Hz, 1H), 7.77 (dd, J = 1.6, 8.8 Hz, 1H), 7.29 (dd, J = 2.0, 8.4 Hz, 1H), 7.25 (d, J = 1.6 Hz, 1H), 6.72 (d, J = 8.4 Hz, 1H), 6.65 (t, J = 6.0 Hz, 1H), 5.70 (q, J = 8.8 Hz, 2H), 4.74 (d, J = 5.6 Hz, 2H), 3.96 (s, 3H), 3.60 (s, 2H), 3.07 (s, 3H), 2.19 (s, 6H).

[0435] Example 222 N-[[4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methyl]-1-methyl-pyrazol-4-amine

[0436] [ka]

[0437]

[0543] The title compound was prepared in a manner similar to that described for Example 221.

[0544] LCMS[M+1, M+3]=452.0, 454.0.

[0438]

[0545] 1H NMR (400 MHz, DMSO-d6) δ = 8.20 (s, 1H), 7.97 (d, J = 8.4 Hz, 1H), 7.75 (dd, J = 1.6, 8.8 Hz, 1H), 7.11 (s, 1H), 7.04 (s, 1H), 5.69 (q, J = 8.8 Hz, 2H), 5.08 (t, J = 6.8 Hz, 1H), 4.37 (d, J = 6.4 Hz, 2H), 3.65 (s, 3H), 3.59 (s, 2H), 2.19 (s, 6H).

[0439] Example 223 4-chloro-8-[(dimethylamino)methyl]-N-[2-(1-methylpyrazol-4-yl)ethyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-amine

[0440] [ka]

[0441]

[0546] Step 1. 8-[(dimethylamino)methyl]-2-[2-(1-methylpyrazol-4-yl)ethylamino]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-4-ol:

[0547] A mixture of 1-[2-chloro-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine (200 mg, 537 μmol, 1.0 equivalent) and 2-(1-methyl-1H-pyrazol-4-yl)ethan-1-amine (560 mg, 4.4 mmol, 8.3 equivalents) was stirred at 120 °C for 2 hours. The reaction was cooled to RT and quenched with HO (10 mL). The mixture was filtered, and the filter cake was washed with HO (30 mL) and dried under vacuum to give the title compound (200 mg, 59% yield) as a brown oil. This material was used in the next step without further purification.

[0442]

[0548] LCMS: [M+1]=448.2.

[0549] Step 2. 4-chloro-8-[(dimethylamino)methyl]-N-[2-(1-methylpyrazol-4-yl)ethyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-amine:

[0550] POCl3 (1.5 mL) was added to a flask containing 8-[(dimethylamino)methyl]-2-[2-(1-methylpyrazol-4-yl)ethylamino]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-4-ol (90.0 mg, 201 μmol, 1.0 equiv.), and the mixture was stirred at 110 °C for 2 h. The reaction mixture was cooled to RT and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 100 mm × 40 mm × 3 μm; mobile phase: 15–45% acetonitrile in water (+0.04% HCl)) to afford the title compound (15.1 mg, 15% yield; hydrochloride salt) as a green solid.

[0443]

[0551] LCMS: [M+1, M+3]=466.2, 468.

[0552] 1 H NMR (400 MHz, DMSO-d6) δ 8.31 (s, 1H), 7.96 - 7.91 (m, 1H), 7.89 - 7.84 (m, 1H), 7.58 (s, 1H), 7.34 (s, 1H), 5.57 (q, J = 8.8 Hz, 2H), 4.44 (br d, J = 5.6 Hz, 2H), 3.78 (s, 3H), 3.51 (br t, J = 7.2 Hz, 2H), 2.75 - 2.69 (m, 8H).

[0444] Example 224 N-benzyl-4-chloro-8-[(dimethylamino)methyl]-N-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-amine

[0445] [ka]

[0446]

[0553] Step 1. 2-[benzyl(methyl)amino]-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-4-ol:

[0554] N-Methyl-1-phenyl-methanamine (2.0 mL, 15.5 mmol, 28.8 equiv.) and 1-[2-chloro-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine (200.0 mg, 536.5 μmol, 1 equiv.) were combined in a flask, and the mixture was stirred at 120° C. for 4 h. The reaction mixture was cooled to RT and quenched with HO (5 mL). The mixture was extracted with EtOAc (3×10 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure to give the title compound (200 mg, 84% yield) as a white solid.

[0447]

[0555] LCMS: [M+H]=444.4.

[0556] Step 2. N-benzyl-4-chloro-8-[(dimethylamino)methyl]-N-methyl-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-amine:

[0557] POCl3 (2 mL) was added to a flask containing 2-[benzyl(methyl)amino]-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-4-ol (200 mg, 451 μmol, 1.0 equiv.), and the mixture was stirred at 110 °C for 2 h. The reaction mixture was cooled to RT, and the pH was adjusted to pH = 9 by adding a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with ethyl acetate (2 × 5 mL). The combined organic layers were washed with water, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Waters Xbridge C18 150 mm × 50 mm × 10 μm; mobile phase: 60–95% acetonitrile in water (+10 mM NH4HCO3)) to afford the title compound (25.0 mg, 12% yield) as a white solid.

[0448]

[0558] LCMS: [M+H, M+3]=462.2, 464.1.

[0559] 1 HNMR (400 MHz, DMSO-d6) δ 8.00 (s, 1H), 7.80 (d, J = 8.4 Hz, 1H), 7.63 (d, J = 8.8 Hz, 1H), 7.37 - 7.19 (m, 5H), 5.52 (q, J = 8.4 Hz, 2H), 4.95 (s, 2H), 3.53 (s, 2H), 3.17 (s, 3H), 2.17 (s, 6H).

[0560]

[0449] [Table 13-1]

[0450] [Table 13-2]

[0451] Example 229 4-chloro-8-[(dimethylamino)methyl]-N-(1-methylpyrazol-4-yl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-amine

[0452] [ka]

[0453]

[0561] Step 1. 8-[(dimethylamino)methyl]-4-methoxy-N-(1-methylpyrazol-4-yl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-amine:

[0562] To a mixture of 1-[2-chloro-4-methoxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-N,N-dimethyl-methanamine (200 mg, 537 μmol, 1 equiv.), 1-methylpyrazol-4-amine (156 mg, 1.61 mmol, 3 equiv.), and CsCO (350 mg, 1.07 mmol, 2 equiv.) in 2-methylbutan-2-ol (2 mL), [2-(2-aminophenyl)phenyl]palladium(II)-dicyclohexyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane methanesulfonate (45 mg, 53.7 μmol, 0.1 equiv.) was added under a nitrogen atmosphere at RT. The mixture was stirred at 90 °C for 12 h. The reaction mixture was cooled to RT and quenched with HO (1 mL). The mixture was extracted with EtOAc (3 × 1 mL). The combined organic layers were washed with brine, dried over NaSO, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO, 10:1 ethyl acetate:methanol) to give the title compound (145 mg, 62% yield) as a yellow solid.

[0454]

[0563] LCMS: [M+H]=434.3.

[0564] Step 2. 4-chloro-8-[(dimethylamino)methyl]-N-(1-methylpyrazol-4-yl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-amine:

[0565] POCl3 (2.0 mL, 21.5 mmol, 71.5 equiv.) was added to a flask containing 8-[(dimethylamino)methyl]-4-methoxy-N-(1-methylpyrazol-4-yl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-amine (130 mg, 300 μmol, 1 equiv.), and the mixture was stirred at 110 °C for 1 h. The mixture was cooled to RT and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 100 mm × 40 mm × 3 μm; mobile phase: 10–40% acetonitrile in water (+0.04% HCl)) to afford the title compound (24.8 mg, 19% yield; hydrochloride salt) as a white solid.

[0455]

[0566] LCMS: [M+1, M+3]=438.2, 440.1.

[0567] 1 H NMR (400 MHz, DMSO-d6) δ 9.77 (s, 1H), 8.52 (br s, 1H), 8.04 (s, 1H), 7.98 (d, J = 8.4 Hz, 1H), 7.86 (d, J = 7.2 Hz, 1H), 7.67 (s, 1H), 5.61 (q, J = 8.8 Hz, 2H), 4.48 (d, J = 1.6 Hz, 2H), 3.88 (s, 3H), 2.76 (s, 6H).

[0568]

[0456] [Table 14]

[0457] Example 233 2-(4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)-N,N-dimethylethan-1-amine

[0458] [ka]

[0459]

[0569] Step 1. 4-Methoxy-5-(2,2,2-trifluoroethyl)-8-vinyl-5H-pyrimido[5,4-b]indole:

[0570] To a solution of 8-iodo-4-methoxy-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole (930 mg, 2.2 mmol, 1.0 equiv.) in dioxane (10.0 mL) and HO (2.0 mL) was added KCO (631 mg, 4.5 mmol, 2.0 equiv.), Pd(dppf)Cl·CHCl (187 mg, 228 μmol, 0.1 equiv.), and 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (7.0 g, 45.6 mmol, 7.7 mL, 20.0 equiv.). The mixture was stirred at 100 °C under a nitrogen atmosphere for 1 h. The mixture was cooled to RT and poured into HO (100 mL). The aqueous phase was extracted with EtOAc (3 × 100 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0-15% ethyl acetate in petroleum ether) to afford the title compound (520 mg, 74% yield) as a white solid.

[0460]

[0571] LCMS[M+1]=308.2.

[0572] 1 H NMR (400 MHz, chloroform-d) δ 8.73 (s, 1H), 8.37 (s, 1H), 7.76 (dd, J = 1.4, 8.6 Hz, 1H), 7.48 (d, J = 8.6 Hz, 1H), 6.91 (dd, J = 10.8, 17.6 Hz, 1H), 5.85 (d, J = 17.6 Hz, 1H), 5.31 (d, J = 10.8 Hz, 1H), 5.15 (q, J = 8.4 Hz, 2H), 4.24 (s, 3H).

[0573] Step 2. 2-(4-Methoxy-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)ethan-1-ol:

[0574] To a solution of 4-methoxy-5-(2,2,2-trifluoroethyl)-8-vinyl-5H-pyrimido[5,4-b]indole (110 mg, 358 μmol, 1.0 equiv.) in THF (1.0 mL) was added 9-BBN (2.1 mL, 3.0 equiv.; 0.5 M solution in THF). The mixture was then stirred at RT for 16 h. The mixture was then cooled to 0° C., and a 1 M aqueous solution of NaOH (220.0 μL, 6.2 equiv.) and HO (241 μL, 2.5 mmol, 7.0 equiv.; 30% solution in water) was added. The mixture was stirred at RT for 30 min. The mixture was quenched with a saturated aqueous solution of NaSO (10 mL) at 0° C. The aqueous phase was extracted with ethyl acetate (3 × 20 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated. The residue was purified by silica gel column chromatography (50-100% ethyl acetate in petroleum ether) to give the title compound (510 mg) as a white solid.

[0461]

[0575] LCMS[M+1]=326.1.

[0576] 1 H NMR (400 MHz, chloroform-d) δ 8.71 (s, 1H), 8.22 (s, 1H), 7.58–7.54 (m, 1H), 7.49–7.45 (m, 1H), 5.14 (q, J = 8.4 Hz, 2H), 4.24 (s, 3H), 3.98 (t, J = 6.6 Hz, 2H), 3.09 (t, J = 6.4 Hz, 3H).

[0577] Step 3. 2-(4-Methoxy-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)acetaldehyde:

[0578] To a solution of 2-(4-methoxy5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)ethan-1-ol (400 mg, 1.2 mmol, 1.0 equiv) in acetonitrile (8.0 mL) was added 2-iodylbenzoic acid (999 mg, 3.5 mmol, 2.9 equiv) at RT. The mixture was stirred at 80 °C for 1 h. The mixture was filtered, and the filtrate was concentrated in vacuo. The residue was purified by silica gel column chromatography (0-25% ethyl acetate in petroleum ether) to afford the title compound (120 mg, 30% yield) as a white solid.

[0462]

[0579] LCMS[M+1]=310.1.

[0580] 1 H NMR (400 MHz, chloroform-d) δ 9.86 (t, J = 2.0 Hz, 1H), 8.75 (s, 1H), 8.27 (br s, 1H), 7.57–7.49 (m, 2H), 5.17 (q, J = 8.4 Hz, 2H), 4.26 (s, 3H), 3.92 (d, J = 2.0 Hz, 3H).

[0581] Step 4. 2-(4-Methoxy-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)-N,N-dimethylethan-1-amine:

[0582] To a solution of 2-(4-methoxy-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)acetaldehyde (100 mg, 309 μmol, 1.0 equiv) in THF (1.5 mL) was added N-methylmethanamine (310 μL, 2.0 equiv; 2 M in THF) and AcOH (35 μL, 619 μmol, 2.0 equiv). The mixture was stirred at RT for 1 h, after which NaBHCN (23 mg, 371 μmol, 1.2 equiv) was added. The mixture was stirred at RT for 1 h. The mixture was poured into HO (30 mL) and extracted with ethyl acetate (3 × 30 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO 2 , 5:1 ethyl acetate:MeOH) to afford the title compound (40.0 mg, 37% yield) as a white solid.

[0463]

[0583] LCMS[M+1]=353.1.

[0584] 1 H NMR (400 MHz, chloroform-d) δ 8.72 (s, 1H), 8.16 (s, 1H), 7.61–7.54 (m, 1H), 7.51–7.45 (m, 1H), 5.14 (q, J = 8.2 Hz, 2H), 4.24 (s, 3H), 3.23–3.13 (m, 2H), 2.93 (br s, 2H), 2.57 (br s, 6H).

[0585] Step 5. 2-(4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)-N,N-dimethylethan-1-amine:

[0586] A solution of 2-(4-methoxy5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)-N,N-dimethylethan-1-amine (40.0 mg, 113.5 μmol, 1.0 equiv) in dichlorophosphorylbenzene (1.5 mL) was stirred at 160° C. for 2 h. The mixture was cooled to RT and quenched with a saturated aqueous solution of NaHCO (50 mL). The mixture was extracted with EtOAc (3×20 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Welch ultimate C18 150 mm×25 mm×7 μm; mobile phase: 10–40% acetonitrile in water (+0.2% formic acid)) to give the title compound (5.3 mg, 13% yield) as a white solid.

[0464]

[0587] LCMS[M+1, M+3]=357.0, 359.0.

[0588] 1 H NMR (400 MHz, chloroform-d) δ 8.93 (s, 1H), 8.53 (br s, 1H), 8.24 (s, 1H), 7.66 (d, J = 8.6 Hz, 1H), 7.53 (d, J = 8.6 Hz, 1H), 5.34 (q, J = 8.0 Hz, 2H), 3.12 - 3.05 (m, 2H), 2.85 - 2.76 (m, 2H), 2.45 (s, 6H).

[0465] Example 234 N-[[4-chloro-8-[(dimethylamino)methyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-2-yl]methyl]-1-methyl-pyrazol-4-amine

[0466] [ka]

[0467]

[0589] The title compound was prepared in a manner similar to that described for Example 233.

[0590] LCMS[M+1, M+3]=412.0, 414.0.

[0468]

[0591] 1 H NMR (400 MHz, chloroform-d) δ 8.92 (s, 1H), 8.22 (s, 1H), 7.62 (br d, J = 8.4 Hz, 1H), 7.51 (d, J = 8.4 Hz, 1H), 5.33 (q, J = 8.0 Hz, 2H), 3.08 - 3.01 (m, 2H), 2.91 - 2.69 (m, 10H), 2.50 (s, 3H).

[0469] Example 235 4-chloro-8-(1-methyl-4-piperidyl)-5-(2,2,2-trifluoroethyl)-pyrimido[5,4-b]indole

[0470] [ka]

[0471]

[0592] Step 1. 8-iodo-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-4-ol:

[0593] To a solution of 4-chloro-8-iodo-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole (1.1 g, 2.67 mmol, 1 equiv.) in dioxane (15 mL) was added an aqueous solution of NaOH (8.02 mL, 3 equiv.; 1 M). The mixture was stirred at 80° C. for 18 hours. The reaction mixture was cooled to RT and poured into water (20 mL). The mixture was extracted with ethyl acetate (3×15 mL), and the combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated in vacuo to give the title compound (1.0 g) as a white solid. This material was used in the next step without further purification.

[0472]

[0594] LCMS[M+1]=394.0.

[0595] 1 H NMR (400 MHz, DMSO-d6) δ 8.34 (d, J = 1.6 Hz, 1H), 8.10 (s, 1H), 7.86 (d, J = 8.8 Hz, 1H), 7.72 (d, J = 8.8 Hz, 1H), 5.64 (q, J = 9.2 Hz, 2H).

[0596] Step 2. 8-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-4-ol:

[0597] To a solution of 8-iodo-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-4-ol (150 mg, 382 μmol, 1 equiv.) and 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine (102 mg, 458 μmol, 1.2 equiv.) in dioxane (1.5 mL) and HO (0.1 mL) was added NaCO (121 mg, 1.14 mmol, 3 equiv.) and Pd(dppf)Cl (28 mg, 38.2 μmol, 0.1 equiv.). The mixture was stirred at 100° C. for 1.5 h. The reaction mixture was cooled to RT and poured into water (10 mL). The mixture was extracted with ethyl acetate (3×10 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150 mm × 25 mm × 10 μm; mobile phase: 1–31% acetonitrile in water (+ formic acid modifier)) to afford the title compound (86 mg, 62% yield) as a white solid.

[0473]

[0598] LCMS[M+1]=363.3.

[0599] 1H NMR (400 MHz, methanol-d4) δ 8.18 (s, 1H), 8.10 (s, 1H), 7.78 (d, J = 8.8 Hz, 1H), 7.68 (d, J = 8.0 Hz, 1H), 6.27 (s, 1H), 5.54 (q, J = 8.8 Hz, 2H), 2.94 - 2.83 (m, 2H), 2.82 - 2.74 (m, 2H), 2.54 - 2.42 (m, 3H), 1.34 - 1.23 (m, 2H).

[0600] Step 3. 8-(1-methylpiperidin-4-yl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-4-ol:

[0601] To a solution of 8-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-4-ol (83.0 mg, 229.0 μmol, 1.0 equiv.) in MeOH (1 mL) was added PtO (260 mg, 1.1 mmol, 5.0 equiv.) under a nitrogen atmosphere. The suspension was degassed under vacuum and purged with H several times. The mixture was stirred under a hydrogen atmosphere (15 psi) at RT for 2 hours. The reaction mixture was filtered and concentrated to give the title compound (65.0 mg, crude) as a brown solid.

[0474]

[0602] LCMS[M+1]=365.2.

[0603] 1 H NMR (400 MHz, chloroform-d) δ 8.05–8.03 (m, 1H), 8.02 (s, 1H), 7.59–7.39 (m, 2H), 5.41–5.33 (m, 2H), 3.07–2.97 (m, 1H), 2.73 (s, 1H), 2.46–2.43 (m, 1H), 2.39–2.32 (m, 2H), 1.98–1.87 (m, 3H), 1.30–1.24 (m, 4H).

[0604] Step 4. 4-Chloro-8-(1-methyl-4-piperidyl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole:

[0605] A solution of 8-(1-methyl-4-piperidyl)-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-4-ol (60 mg, 165 μmol, 1 equiv.) in POCl3 (0.5 mL) was stirred at 110 °C for 16 h. The reaction mixture was cooled to RT and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150 mm × 25 mm × 10 μm; mobile phase: 0–27% acetonitrile in water (+ formic acid modifier)) to afford the title compound (10.4 mg, 17% yield) as a yellow solid.

[0475]

[0606] LCMS[M+1, M+3]=383.0, 384.9.

[0607] 1 H NMR (400 MHz, chloroform-d) δ 8.92 (s, 1H), 8.26 (s, 1H), 7.68 (d, J = 8.4 Hz, 1H), 7.53 (d, J = 8.4 Hz, 1H), 5.40 - 5.27 (m, 2H), 3.07 (d, J = 11.2 Hz, 2H), 2.82 - 2.66 (m, 1H), 2.39 (s, 3H), 2.24 - 2.11 (m, 2H), 1.97 (s, 4H).

[0476] Example 236 and Example 237 4-chloro-8-[(3R)-1-methyl-3-piperidyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole and 4-chloro-8-[(3S)-1-methyl-3-piperidyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole

[0477] [ka]

[0478]

[0608] Step 1. tert-Butyl 5-[4-hydroxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-3,6-dihydro-2H-pyridine-1-carboxylate:

[0609] To a solution of tert-butyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-1-carboxylate (189 mg, 611 μmol, 1.2 equiv.) and 8-iodo-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-4-ol (200 mg, 509 μmol, 1.0 equiv.) in dioxane (2.0 mL) and HO (0.4 mL), NaCO (161.7 mg, 1.5 mmol, 3.0 equiv.) and Pd(dppf)Cl (37.2 mg, 50.9 μmol, 0.1 equiv.) were added under a nitrogen atmosphere. The mixture was stirred at 100° C. for 1 hour. The mixture was cooled to RT and poured into water (5 mL). The mixture was extracted with ethyl acetate (3 × 5 mL). The combined organic layers were washed with brine, dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0–100% ethyl acetate in petroleum ether) to afford the title compound (350 mg, 63% yield) as a yellow solid.

[0479]

[0610] LCMS[M+1]=449.1.

[0611] 1 H NMR (400 MHz, DMSO-d6) δ 12.62 (s, 1H), 8.09 (d, J = 2.4 Hz, 1H), 7.95 (s, 1H), 7.83 (d, J = 8.8 Hz, 1H), 7.73 (d, J = 8.8 Hz, 1H), 6.37 (s, 1H), 5.64 (q, J = 8.8 Hz, 2H), 4.32 (s, 2H), 3.50 (t, J = 4.8 Hz, 2H), 2.34 - 2.26 (s, 2H), 1.44 (s, 9H).

[0612] Step 2. tert-Butyl 3-(4-hydroxy-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)piperidine-1-carboxylate:

[0613] 10% Pd on C (831 mg, 781 μmol, 1.0 equiv.) was added to the reaction flask under argon, and methanol (2.0 mL) was added to quench the catalyst. A solution of tert-butyl 5-[4-hydroxy-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-8-yl]-3,6-dihydro-2H-pyridine-1-carboxylate (350 mg, 781 μmol, 1.0 equiv.) in methanol (4.0 mL) was then added. The suspension was degassed under vacuum and purged with hydrogen gas three times. The mixture was stirred under a hydrogen atmosphere at 40° C. for 12 hours. The reaction mixture was filtered and concentrated to give the title compound (300 mg, 75% yield) as a yellow solid.

[0480]

[0614] LCMS[M+1]=451.1.

[0615] 1 H NMR (400 MHz, methanol-d4) δ 8.09 (s, 1H), 8.04 (s, 1H), 7.67 (d, J = 8.8 Hz, 1H), 7.56 (d, J = 9.2 Hz, 1H), 5.53 (q, J = 8.8 Hz, 2H), 4.22 - 4.13 (m, 2H), 2.92 - 2.85 (m, 3H), 2.10 (d, J = 10.4 Hz, 1H), 1.85 (d, J = 11.2 Hz, 2H), 1.68 - 1.58 (m, 1H), 1.48 (s, 9H).

[0616] Step 3. 8-(piperidin-3-yl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-4-ol:

[0617] HCl (3.0 mL, 18.0 equiv.; 4 M solution in dioxane) was added to a flask containing tert-butyl 3-(4-hydroxy-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)piperidine-1-carboxylate (300 mg, 666 μmol, 1.0 equiv.). The mixture was stirred at RT for 30 min. The reaction mixture was concentrated under reduced pressure to provide the title compound (230 mg, 80% yield; hydrochloride salt) as a white solid.

[0481]

[0618] LCMS[M+1]=351.2.

[0619] 1 H NMR (400 MHz, methanol-d4) δ 8.15 (s, 1H), 7.97 (s, 1H), 7.47 (d, J = 8.4 Hz, 1H), 7.36 (d, J = 8.4 Hz, 1H), 5.59 - 5.53 (m, 2H), 3.17 - 3.06 (m, 2H), 2.88 - 2.82 (m, 1H), 2.73 - 2.60 (m, 2H), 2.05 (d, J = 10.8 Hz, 1H), 1.86 - 1.65 (m, 4H).

[0620] Step 4. 8-[(3R)-1-Methyl-3-piperidyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-4-ol and 8-[(3S)-1-Methyl-3-piperidyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-4-ol:

[0621] To a solution of 8-(piperidin-3-yl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-4-ol (230 mg, 657 μmol, 1.0 equiv.) in DCM (11.5 mL) and methanol (9.2 mL) was added formaldehyde (145 μL, 5.2 mmol, 8.0 equiv.; 37% solution in water) at RT. After 1 h, NaBH(OAc) (1.1 g, 5.6 mmol, 8.0 equiv.) was added, and the mixture was stirred at RT for 2 h. A saturated aqueous solution of NaHCO was added, and the mixture was extracted with ethyl acetate (3 × 20 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative SFC (column: DAICEL CHIRALPAK IG (250 mm × 30 mm × 10 μm); mobile phase: 30% methanol in CO (+0.1% NH H O)) to give 8-[(3R)-1-methyl-3-piperidyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-4-ol (85.0 mg, 35% yield).

[0482]

[0622] LCMS[M+1]=365.1.

[0623] 1 H NMR (400 MHz, methanol-d4) δ 8.08 (s, 1H), 8.02 (s, 1H), 7.65 (d, J = 8.8 Hz, 1H), 7.54 (dd, J = 1.6, 8.8 Hz, 1H), 5.52 (q, J = 8.8 Hz, 2H), 3.11 - 2.98 (m, 3H), 2.36 (s, 3H), 2.20 (t, J = 11.2 Hz, 1H), 2.12 (dt, J = 2.4, 12.0 Hz, 1H), 2.04 - 2.00 (m, 1H), 1.93 - 1.87 (m, 1H), 1.86 - 1.75 (m, 1H), 1.66 - 1.56 (m, 1H).

[0624] and 8-[(3S)-1-methyl-3-piperidyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-4-ol (88.0 mg, 36% yield).

[0483]

[0625] LCMS[M+1]=365.1.

[0626] 1 H NMR (400 MHz, methanol-d4) δ 8.08 (s, 1H), 8.03 (s, 1H), 7.66 (d, J = 8.8 Hz, 1H), 7.54 (dd, J = 1.6, 8.8 Hz, 1H), 5.52 (q, J = 8.8 Hz, 2H), 3.14 - 2.98 (m, 3H), 2.39 (s, 3H), 2.27 - 2.13 (m, 2H), 2.06 - 2.01 (m, 1H), 1.95 - 1.76 (m, 2H), 1.68 - 1.57 (m, 1H).

[0627] Step 5. 4-chloro-8-[(3R)-1-methyl-3-piperidyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indole:

[0628] POCl3 (1.0 mL) was added to a flask containing 8-[(3R)-1-methyl-3-piperidyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-4-ol (50.0 mg, 137 μmol, 1.0 equiv.), and the mixture was stirred at 110 °C for 2 h. The reaction mixture was cooled to RT and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150 mm × 25 mm × 10 μm; mobile phase: 12–42% acetonitrile in water (+ formic acid modifier)) to afford the title compound (22.5 mg, 42% yield; formate salt) as a white solid.

[0484]

[0629] LCMS[M+1, M+3]=383.0, 385.0.

[0630] 1H NMR (400 MHz, methanol-d4) δ 8.88 (s, 1H), 8.49 (s, 1H), 8.32 (s, 1H), 7.90 - 7.87 (m, 1H), 7.80 (dd, J = 1.6, 8.8 Hz, 1H), 5.63 (q, J = 8.4 Hz, 2H), 3.62 - 3.53 (m, 2H), 3.27 - 3.26 (m, 1H), 3.18 - 3.12 (m, 1H), 3.03 - 2.97 (m, 1H), 2.89 (s, 3H), 2.14 (d, J = 11.6 Hz, 2H), 2.05 - 1.84 (m, 2H).

[0631] Step 5. 4-Chloro-8-[(3S)-1-methyl-3-piperidyl]-5-(2,2,2-trifluoroethyl)-pyrimido[5,4-b]indole:

[0632] POCl3 (1.0 mL) was added to a flask containing 8-[(3S)-1-methyl-3-piperidyl]-5-(2,2,2-trifluoroethyl)pyrimido[5,4-b]indol-4-ol (50.0 mg, 137.2 μmol, 1.0 equiv.), and the mixture was stirred at 110 °C for 2 h. The reaction mixture was cooled to RT and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150 mm × 25 mm × 10 μm; mobile phase: 12–42% acetonitrile in water (+ formic acid modifier)) to afford the title compound (18.8 mg, 35% yield; formate salt) as a white solid.

[0485]

[0633] LCMS[M+1, M+3]=383.0, 385.0.

[0634] 1H NMR (400 MHz, methanol-d4) δ 8.89 (s, 1H), 8.48 (s, 1H), 8.32 (s, 1H), 7.90 - 7.87 (m, 1H), 7.81 - 7.79 (m, 1H), 5.63 (q, J = 8.4 Hz, 2H), 3.62 - 3.52 (m, 2H), 3.29 - 3.25 (m, 1H), 3.17 - 3.11 (m, 1H), 2.99 (dt, J = 2.4, 12.4 Hz, 1H), 2.88 (s, 3H), 2.14 (d, J = 11.4 Hz, 2H), 2.04 - 1.84 (m, 2H).

[0486] Example 238 and Example 239 (R)-4-chloro-8-(1-(4-methylpiperazin-1-yl)ethyl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole and (S)-4-chloro-8-(1-(4-methylpiperazin-1-yl)ethyl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole

[0487] [ka]

[0488]

[0635] Step 1. 1-(4-Methoxy-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)ethan-1-one:

[0636] To a mixture of 8-iodo-4-methoxy-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole (500 mg, 1.2 mmol, 1.0 equiv.) and tributyl(1-ethoxyvinyl)stannane (830 μL, 2.4 mmol, 2.0 equiv.) in toluene (5.0 mL) was added Pd(PPh3)4 (141.9 mg, 122.8 μmol, 0.1 equiv.) under a nitrogen atmosphere at RT. The reaction mixture was stirred at 80 °C for 2 h. The mixture was cooled to RT and poured into a saturated aqueous solution of KF (15 mL). The mixture was then extracted with ethyl acetate (3 × 15 mL). The combined organic extracts were washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0-100% ethyl acetate in petroleum ether) to afford the title compound (350 mg, 79% yield) as a yellow solid.

[0489]

[0637] LCMS: [M+1]=324.1.

[0638] 1 H NMR (400 MHz, methanol-d4) δ 9.13 (s, 1H), 9.03 (s, 1H), 8.47 (s, 1H), 8.03 (d, J = 9.2 Hz, 1H), 5.58 (q, J = 8.4 Hz, 2H), 4.45 (s, 3H), 2.76 (s, 3H).

[0639] Step 2. 4-Methoxy-8-(1-(4-methylpiperazin-1-yl)ethyl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole:

[0640] To a solution of 1-methylpiperazine (137 μL, 1.2 mmol, 2.0 equiv) in titanium(IV) isopropoxide (2.0 mL) was added 1-(4-methoxy-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indol-8-yl)ethan-1-one (200 mg, 619 μmol, 1.0 equiv) at RT. The mixture was stirred at 90° C. for 1 h, and then NaBHCN (311 mg, 4.9 mmol, 8.0 equiv) was added. The mixture was stirred at 90° C. for 12 h. The reaction mixture was cooled to RT, and the mixture was diluted with water and ethyl acetate. The filtrate was extracted with ethyl acetate (5×15 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (10:1 EtOAc:MeOH) to afford the title compound (170 mg, 65% yield) as a yellow oil.

[0490]

[0641] LCMS: [M+1]=408.3.

[0642] The racemic title compound was separated by SFC (column: DAICEL CHIRALPAK IG (250 mm × 30 mm × 10 μm); mobile phase: 35% methanol in CO (+0.1% NH·H O)) to give (R)-4-methoxy 8-(1-(4-methylpiperazin-1-yl)ethyl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole (80.0 mg, 45% yield):

[0643] 1H NMR (400 MHz, chloroform-d) δ 8.72 (s, 1H), 8.26 (s, 1H), 7.67 (dd, J = 1.6, 8.8 Hz, 1H), 7.47 (d, J = 8.8 Hz, 1H), 5.20 - 5.08 (m, 2H), 4.24 (s, 3H), 3.61 (d, J = 6.8 Hz, 1H), 2.73 - 2.31 (m, 8H), 2.26 (s, 3H), 1.47 (d, J = 6.8 Hz, 3H). and (S)-4-methoxy-8-(1-(4-methylpiperazin-1-yl)ethyl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole (60.0 mg, 34% yield):

[0644] 1 H NMR (400 MHz, chloroform-d) δ 8.73 (s, 1H), 8.30 (s, 1H), 7.66–7.59 (m, 1H), 7.54 (d, J = 8.0 Hz, 1H), 5.18 (q, J = 8.4 Hz, 2H), 4.25 (s, 3H), 3.94–3.72 (m, 1H), 3.47–2.85 (m, 8H), 2.72 (d, J = 18.4 Hz, 3H), 1.26 (s, 3H).

[0645] Step 3. (R)-4-chloro-8-(1-(4-methylpiperazin-1-yl)ethyl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole:

[0646] Phenylphosphonic acid dichloride (0.8 mL) was added to a flask containing (R)-4-methoxy-8-(1-(4-methylpiperazin-1-yl)ethyl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole (80.0 mg, 196.3 μmol, 1.0 equiv.), and the mixture was stirred at 140° C. for 2 h. The mixture was cooled to RT and poured into a saturated aqueous solution of NaHCO. The pH was adjusted to 7 by adding solid NaHCO. The aqueous phase was extracted with ethyl acetate (5×15 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150 x 25 mm x 10 um; mobile phase: 10-40% acetonitrile in water (+ formic acid modifier)) to give the title compound (6.8 mg, 8% yield) as a colorless oil.

[0491]

[0647] LCMS: [M+1, M+3]=412.0, 414.0.

[0648] 1 H NMR (400 MHz, chloroform-d) δ 8.93 (s, 1H), 8.36 (s, 1H), 7.75 (dd, J = 1.2, 8.8 Hz, 1H), 7.54 (d, J = 8.8 Hz, 1H), 5.35 (q, J = 8.4 Hz, 2H), 3.72 - 3.68 (m, 1H), 2.86 (s, 5H), 2.66 (d, J = 5.6 Hz, 3H), 2.53 (s, 3H), 1.47 (d, J = 6.8 Hz, 3H).

[0649] Step 3. (S)-4-Chloro-8-(1-(4-methylpiperazin-1-yl)ethyl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole:

[0650] Phenylphosphonic acid dichloride (0.8 mL) was added to a flask containing (S)-4-methoxy8-(1-(4-methylpiperazin-1-yl)ethyl)-5-(2,2,2-trifluoroethyl)-5H-pyrimido[5,4-b]indole (60.0 mg, 147.2 μmol, 1.0 equiv.), and the mixture was stirred at 140° C. for 2 h. The mixture was cooled to RT and poured into a saturated aqueous solution of NaHCO. The pH was adjusted to pH=7 by adding solid NaHCO. The aqueous phase was extracted with ethyl acetate (5×15 mL). The combined organic layers were washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150 mm × 25 mm × 10 μm; mobile phase: 10-40% acetonitrile in water (+ formic acid modifier)) to give the title compound (6.6 mg, 10% yield) as a colorless oil.

[0492]

[0651] LCMS: [M+1, M+3]=412.3, 414.3.

[0652] 1 H NMR (400 MHz, chloroform-d) δ 8.94 (s, 1H), 8.35 (s, 1H), 7.73 (d, J = 8.4 Hz, 1H), 7.54 (d, J = 8.4 Hz, 1H), 5.34 (q, J = 8.0 Hz, 2H), 3.70 (d, J = 4.8 Hz, 1H), 3.13 - 2.51 (m, 11H), 1.45 (d, J = 6.0 Hz, 3H).

[0653] Example 240 2-[4-chloro-8-[(dimethylamino)methyl]pyrimido[5,4-b]indol-5-yl]acetonitrile

[0493] [ka]

[0494]

[0654] Step 1. (4-chloro-5H-pyrimido[5,4-b]indol-8-yl)methanol:

[0655] To a solution of methyl 4-chloro-5H-pyrimido[5,4-b]indole-8-carboxylate (1.3 g, 4.97 mmol, 1.0 equiv) in THF (13.0 mL) was added LiAlH (377 mg, 9.9 mmol, 2.0 equiv) under a nitrogen atmosphere at 0 °C. The mixture was stirred at RT for 2 h. The reaction mixture was cooled to 0 °C and quenched by the sequential addition of HO (1.3 mL), 15% aqueous NaOH (1.3 mL), and HO (3.9 mL). After stirring at RT for 10 min, the solid was removed by filtration. The filtrate was concentrated, and the residue was purified by silica gel column chromatography (10:1 DCM:methanol) to afford the title compound (800 mg, 69% yield) as a yellow solid.

[0495]

[0656] LCMS: [M+1, M+3]=234.1, 236.1.

[0657] 1 H NMR (400 MHz, methanol-d4) δ 8.79 (s, 1H), 8.29 (d, J = 0.8 Hz, 1H), 7.76–7.71 (m, 1H), 7.67–7.63 (m, 1H), 4.79 (s, 2H).

[0658] Step 2. 4-Chloro-5H-pyrimido[5,4-b]indole-8-carbaldehyde:

[0659] A mixture of (4-chloro-5H-pyrimido[5,4-b]indol-8-yl)methanol (200.0 mg, 0.86 mmol, 1.0 equiv) and MnO (744.1 mg, 8.5 mmol, 10 equiv) in DCE (6.0 mL) was degassed with nitrogen, and then the mixture was stirred under a nitrogen atmosphere at 70 °C for 1 h. The reaction mixture was filtered and concentrated under reduced pressure to give the title compound (120 mg, 61% yield) as a yellow solid.

[0496]

[0660] LCMS: [M+1, M+3]=232.1, 234.1.

[0661] 1 H NMR (400 MHz, DMSO-d6) δ 12.96 - 12.86 (m, 1H), 10.15 (s, 1H), 8.96 (s, 1H), 8.88 (s, 1H), 8.20 (br d, J = 8.4 Hz, 1H), 7.83 (d, J = 8.4 Hz, 1H).

[0662] Step 3. 2-(4-chloro-8-formyl-pyrimido[5,4-b]indol-5-yl)acetonitrile:

[0663] 4-Chloro-5H-pyrimido[5,4-b]indole-8-carbaldehyde (100 mg, 432 μmol, 1.0 equiv), 2-iodoacetonitrile (79 mg, 475 μmol, 1.1 equiv), and KCO (179 mg, 1.3 mmol, 3.0 equiv) were combined in DMF (1.0 mL), and the mixture was stirred at 50 °C for 3 h. The reaction mixture was poured into HO (5 mL) and extracted with ethyl acetate (3 × 10 mL). The organic layer was washed with brine, dried over NaSO, and concentrated under reduced pressure to give the title compound (60 mg, 51% yield) as a white solid.

[0497]

[0664] LCMS: [M+1, M+3]=271.1, 273.1.

[0665] 1 H NMR (400 MHz, DMSO-d6) δ 10.28 - 10.12 (m, 1H), 9.05 (s, 1H), 8.94 (s, 1H), 8.36 (d, J = 9.2. Hz, 1H), 8.25 (d, J = 9.2 Hz, 1H), 6.07 (s, 2H).

[0666] Step 4. 2-[4-chloro-8-[(dimethylamino)methyl]pyrimido[5,4-b]indol-5-yl]acetonitrile:

[0667] A solution of 2-(4-chloro-8-formyl-pyrimido[5,4-b]indol-5-yl)acetonitrile (40.0 mg, 148 μmol, 1.0 equiv.), dimethylamine (81 μL, 1.1 equiv.; 2.0 M in THF), and AcOH (17 μL, 296 μmol, 2.0 equiv.) in DCE (2.0 mL) was stirred at RT for 30 min. NaBH(OAc)3 (78.3 mg, 369 μmol, 2.5 equiv.) was then added, and the mixture was stirred at RT for 2 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150 mm × 25 mm × 10 μm; mobile phase: 0–27% acetonitrile in water (+ formic acid modifier)) to afford the title compound (13.7 mg, 30% yield) as a yellow solid.

[0498]

[0668] LCMS: [M+1, M+3]=300.1, 302.0.

[0669] 1 H NMR (400 MHz, DMSO-d6) δ 8.94 (s, 1H), 8.21 (s, 1H), 8.02 (d, J = 8.8 Hz, 1H), 7.82 (d, J = 8.8 Hz, 1H), 5.99 (s, 2H), 3.63 (s, 2H), 2.20 (s, 6H). Preparation of Y220C TP53 TR-FRET binding probe, BP1. 5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]-N-[2-[2-[2-[[3-methoxy-4-[3-[4-(tetrahydropyran-4-ylamino)-1-(2,2,2-trifluoroethyl)indol-2-yl]prop-2-ynylamino]phenyl]sulfonylamino]ethoxy]ethoxy]ethoxy]ethyl]pentanamide

[0499] [ka]

[0500]

[0670] BP1 synthesis, step 1a. 2-iodo-4-nitro-1-(2,2,2-trifluoroethyl)indole: To a solution of 2-iodo-4-nitro-1H-indole (3.0 g, 10.4 mmol, 1.0 equiv) in THF (20 mL), NaH (2.0 g, 52.0 mmol, 5.0 equiv; 60.0% dispersion in oil) was added portionwise at 0 °C and stirred at 0 °C for 30 min. 2,2,2-Trifluoroethyl trifluoromethanesulfonate (9.6 g, 41.0 mmol, 4.0 equiv) was added portionwise to the reaction mixture at 0 °C. The mixture was stirred at 25 °C for 2 h. The reaction mixture was quenched with HO (20 mL), and the resulting mixture was partitioned between EtOAc (200 mL) and HO (200 mL), and the aqueous layer was extracted with EtOAc (2 × 200 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (15:1 to 12:1 petroleum ether:EtOAc) to afford the title compound (5.0 g, crude) as a yellow solid.

[0501]

[0671] 1 H NMR (400 MHz, chloroform-d) δ 8.15 (d, J = 8.2 Hz, 1H), 7.80–7.63 (m, 2H), 7.33 (t, J = 8.2 Hz, 1H), 4.85 (q, J = 8.2 Hz, 2H).

[0672] BP1 synthesis, step 2a. 2-iodo-1-(2,2,2-trifluoroethyl)indol-4-amine: To a solution of 2-iodo-4-nitro-1-(2,2,2-trifluoroethyl)indole (1.9 g, 5.1 mmol, 1.0 equiv) in EtOH (20.0 mL) and HO (5.0 mL) was added Fe (717 mg, 12.8 mmol, 2.5 equiv) and NHCl (687 mg, 12.8 mmol, 2.5 equiv). The mixture was stirred at 80 °C for 2 h. The reaction solution was filtered through a pad of diatomaceous earth, and the filtrate was partitioned between EtOAc (200 mL) and HO (200 mL). The aqueous layer was extracted with EtOAc (2 × 200 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to give the title compound (1.7 g, 97% yield) as a yellow solid.

[0502]

[0673] LCMS[M+1]=341.1.

[0674] BP1 synthesis, step 3a. 2-iodo-N-tetrahydropyran-4-yl-1-(2,2,2-trifluoroethyl)indol-4-amine: To a solution of 2-iodo-1-(2,2,2-trifluoroethyl)indol-4-amine (600 mg, 1.7 mmol, 1.0 equiv) in DMF (10 mL) was added chloro(trimethyl)silane (559.8 μL, 4.4 mmol, 2.5 equiv) and tetrahydropyran-4-one (648.1 μL, 7.0 mmol, 4.0 equiv). The mixture was stirred at 0°C for 2 h. Borane-tetrahydrofuran complex (1 M, 8.8 mL, 5.0 equiv) was added to the mixture under N2, and the resulting mixture was stirred at 0-20°C for 12 h. The reaction mixture was poured into a saturated aqueous solution of NH4Cl (1.5 mL), extracted with EtOAc (3 × 5 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the title compound (600 mg, 80% yield) as a white solid.

[0503]

[0675] 1H NMR (400 MHz, chloroform-d) δ 8.03 (s, 1H), 7.07 (t, J = 8.0 Hz, 1H), 6.90 - 6.70 (m, 2H), 6.34 (d, J = 7.8 Hz, 1H), 4.69 (q, J = 8.4 Hz, 2H), 4.11 - 4.00 (m, 2H), 3.92 - 3.71 (m, 1H), 3.73 - 3.63 (m, 1H), 3.56 (t, J = 10.6 Hz, 2H), 2.97 (s, 2H), 2.89 (s, 1H), 2.68 (d, J = 9.4Hz, 1H), 2.24 - 2.01 (m, 2H).

[0676] LCMS[M+1]=425.1.

[0504]

[0677] BP1 synthesis, step 1b. tert-butyl N-[2-[2-[2-[2-[(3-methoxy-4-nitro-phenyl)sulfonylamino]ethoxy]ethoxy]-ethoxy]ethyl]carbamate: To a solution of tert-butyl N-[2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]-ethyl]carbamate (906 mg, 3.1 mmol, 1.3 equiv) in DCM (15 mL) was added TEA (1.6 mL, 11.9 mmol, 5.0 equiv) and 3-methoxy-4-nitro-benzenesulfonyl chloride (600 mg, 2.4 mmol, 1.0 equiv). The mixture was stirred at 15 °C for 1 h. The reaction mixture was concentrated under reduced pressure, and the resulting residue was partitioned between EtOAc (50 mL) and HO (30 mL), and the aqueous layer was extracted with EtOAc (2 × 50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to give the title compound (1.1 g, 91% yield) as a brown oil.

[0505]

[0678] 1H NMR (400 MHz, DMSO-d6) δ 8.07 (d, J = 8.4 Hz, 1H), 7.67 (s, 1H), 7.51 (d, J = 8.4 Hz, 1H), 6.75 (s, 1H), 4.00 (s, 3H), 3.53 - 3.41 (m, 10H), 3.15 - 2.90 (m, 6H), 2.65 (t, J = 5.7 Hz, 1H), 1.36 (s, 9H).

[0679] LCMS[M+1]=408.2.

[0506]

[0680] BP1 synthesis, step 2b. tert-butyl N-[2-[2-[2-[2-[(4-amino-3-methoxyphenyl)sulfonylamino]ethoxy]ethoxy]-ethoxy]ethyl]carbamate: To a solution of tert-butyl N-[2-[2-[2-[2-[(3-methoxy-4-nitrophenyl)sulfonylamino]ethoxy]ethoxy]ethoxy]-ethyl]carbamate (2.0 g, 3.9 mmol, 1.0 equiv.) in EtOH (12.0 mL) and HO (3.0 mL) was added Fe (1.1 g, 19.7 mmol, 5.0 equiv.) and NH4Cl (1.1 g, 19.7 mmol, 5.0 equiv.). The mixture was stirred at 80 °C for 1 h. The suspension was filtered through a pad of Celite, and the pad cake was washed with EtOH (3 × 20 mL). The filtrate was concentrated under reduced pressure, and the resulting residue was partitioned between EtOAc (50 mL) and HO (30 mL). The aqueous layer was then re-extracted with EtOAc (2 × 50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure to give the title compound (1.5 g, 80% yield) as a brown oil.

[0507]

[0681] 1H NMR (400 MHz, DMSO-d6) δ 7.20 (t, J = 6.0 Hz, 1H), 7.16 - 7.13 (m, 1H), 7.12 (s, 1H), 6.76 (br t, J = 5.6 Hz, 1H), 6.69 - 6.63 (m, 1H), 6.73 - 6.60 (m, 1H), 5.56 (s, 2H), 3.80 (s, 3H), 3.50 - 3.42 (m, 7H), 3.40 - 3.35 (m, 3H), 3.05 (q, J = 6.0 Hz, 2H), 2.80 (q, J = 6.0 Hz, 2H), 1.36 (s, 9H).

[0682] LCMS[M+1]=378.3.

[0508]

[0683] BP1 synthesis, step 3b. tert-butyl N-[2-[2-[2-[2-[[3-methoxy4-(3-trimethylsilylprop-2-ynylamino)phenyl]-sulfonylamino]ethoxy]ethoxy]ethoxy]ethyl]carbamate: To a solution of tert-butyl N-[2-[2-[2-[2-[(4-amino-3-methoxyphenyl)sulfonylamino]ethoxy]ethoxy]ethoxy]-ethyl]carbamate (1.5 g, 3.1 mmol, 1.0 equiv) in DCM (15 mL) and acetic acid (3 mL) was added 3-trimethylsilylprop-2-ynal (396 mg, 3.1 mmol, 1.0 equiv). The mixture was stirred at 35° C. for 17 hours, and then sodium triacetoxyborohydride (2.6 g, 12.5 mmol, 4.0 equiv) was added to the mixture. The mixture was stirred at 35° C. for 17 hours. The reaction mixture was diluted with EtOAc (50 mL) and HO (30 mL), and the aqueous layer was extracted with EtOAc (2×50 ml). The combined organic extracts were washed with brine (50 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative HPLC (column: Phenomenex Luna C18 (250 mm × 70 mm × 15 μm); mobile phase: 40-75% ACN in water (+NHHCO modifier)) to afford the title compound (480 mg, 26% yield) as a white solid.

[0509]

[0684] 1 H NMR (400 MHz, DMSO-d6) δ 7.33 - 7.23 (m, 2H), 7.16 (d, J = 1.8 Hz, 1H), 6.73 (br t, J = 5.2 Hz, 1H), 6.68 (d, J = 8.4 Hz, 1H), 6.11 (t, J = 6.0 Hz, 1H), 4.01 (d, J = 6.0 Hz, 2H), 3.83 (s, 3H), 3.54 - 3.40 (m, 8H), 3.36 (br t, J = 5.8 Hz, 4H), 3.31 (s, 2H), 3.05 (q, J = 6.0 Hz, 2H), 2.82 (q, J = 6.0 Hz, 2H), 2.07 (s, 1H), 1.36 (s, 9H).

[0685] LCMS[M+1]=488.4.

[0510]

[0686] BP1 synthesis, step 4b. tert-butyl N-[2-[2-[2-[2-[[3-methoxy-4-(prop-2-ynylamino)phenyl]sulfonylamino]-ethoxy]ethoxy]ethoxy]ethyl]carbamate: To a solution of tert-butyl N-[2-[2-[2-[2-[[3-methoxy-4-(3-trimethylsilylprop-2-ynylamino)phenyl]sulfonylamino]ethoxy]ethoxy]ethoxy]ethyl]-carbamate (580.0 mg, 986.7 μmol, 1.0 equiv) in MeOH (6.0 mL) was added KCO (272.7 mg, 1.9 mmol, 2.0 equiv). The mixture was stirred at 20 °C for 1 h. The reaction mixture was partitioned between EtOAc (50 mL) and HO (30 mL), and the aqueous layer was extracted with EtOAc (2 × 50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to afford the title compound (400 mg, 79% yield) as an off-white oil.

[0511]

[0687] 1 H NMR (400 MHz, DMSO-d6) δ 7.34 - 7.24 (m, 2H), 7.15 (d, J = 1.8 Hz, 1H), 6.74 (t, J = 5.4 Hz, 1H), 6.69 (d, J = 8.4 Hz, 1H), 6.13 (t, J = 6.0 Hz, 1H), 3.97 (dd, J = 2.0, 6.2 Hz, 2H), 3.83 (s, 3H), 3.50 - 3.41 (m, 8H), 3.39 - 3.33 (m, 4H), 3.11 - 3.00 (m, 3H), 2.81 (q, J = 6.0 Hz, 2H), 2.07 (s, 3H), 1.36 (s, 9H).

[0688] LCMS[M+1]=416.3.

[0512]

[0689] BP1 synthesis, step 5b. tert-butyl N-[2-[2-[2-[2-[[3-methoxy-4-[3-[4-(tetrahydropyran-4-ylamino)-1-(2,2,2-trifluoroethyl)indol-2-yl]prop-2-ynylamino]phenyl]sulfonylamino]ethoxy]ethoxy]ethoxy]-ethyl]carbamate: To a solution of tert-butyl N-[2-[2-[2-[2-[[3-methoxy-4-(prop-2-ynylamino)phenyl]sulfonylamino]ethoxy]-ethoxy]ethoxy]ethyl]carbamate (200 mg, 388 μmol, 1.0 equiv.) in ACN (2.0 mL), dichloropalladium-triphenylphosphane (27.2 mg, 38.7 μmol, 0.1 equiv.) and copper(I) iodide (7.4 mg, 38.8 μmol, 0.1 equiv.), 2-iodo-N-tetrahydropyran-4-yl-1-(2,2,2-trifluoroethyl)indol-4-amine (165 mg, 388 μmol, 1.0 equiv.) and triethylamine (162 μL, 1.1 mmol, 3.0 equiv.) were added under a nitrogen atmosphere. The mixture was stirred at 70° C. for 2 hours. The reaction mixture was partitioned between EtOAc (10 mL) and HO (10 mL), and the aqueous layer was extracted with EtOAc (2×10 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to give the title compound (200 mg, 64% yield) as a white solid.

[0513]

[0690] 1H NMR (400 MHz, DMSO-d6) δ 7.47 (s, 1H), 7.32 - 7.24 (m, 1H), 7.19 (d, J = 1.6 Hz, 2H), 7.08 (s, 1H), 7.00 (t, J = 8.0 Hz, 1H), 6.83 (d, J = 8.4 Hz, 1H), 6.80 - 6.63 (m, 2H), 6.32 (t, J = 6.4 Hz, 1H), 6.21 (d, J = 7.8 Hz, 1H), 5.96 - 5.96 (m, 1H), 5.54 (d, J = 8.4 Hz, 1H), 5.01 - 4.84 (m, 2H), 4.33 (d, J = 6.2 Hz, 1H), 3.93 - 3.83 (m, 4H), 3.51 - 3.39 (m, 9H), 3.38 - 3.34 (m, 1H), 3.38 - 3.29 (m, 7H), 3.04 (q, J = 5.8 Hz, 2H), 2.81 (q, J = 5.8 Hz, 2H), 2.50 (d, J = 1.6 Hz, 72H), 2.04 - 1.97 (m, 1H), 1.91 (br d, J = 13.4 Hz, 1H), 1.95 - 1.85 (m, 1H), , 1.36 (s, 9H).

[0691] LCMS[M+1]=812.3.

[0514]

[0692] BP1 synthesis, step 6b. N-[2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]ethyl]-3-methoxy-4-[3-[4-(tetrahydropyran-4-ylamino)-1-(2,2,2-trifluoroethyl)indol-2-yl]prop-2-ynylamino]benzenesulfonamide: To a solution of tert-butyl N-[2-[2-[2-[2-[[3-methoxy 4-[3-[4-(tetrahydropyran-4-ylamino)-1-(2,2,2-trifluoroethyl)indol-2-yl]prop-2-ynylamino]phenyl]sulfonylamino]ethoxy]ethoxy]ethoxy]-ethyl]carbamate (200 mg, 246 μmol, 1.0 equiv) in DCM (0.6 mL) was added TFA (0.8 mL). The mixture was stirred at 20° C. for 1 hour. The reaction mixture was concentrated under reduced pressure to give the title compound (150 mg, 86% yield) as a brown oil. The crude product was used directly in the next step.

[0515]

[0693] LCMS[M+1]=712.4.

[0694] BP1 synthesis, step 7b, TR-FRET conjugated probe, BP1. 5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]-N-[2-[2-[2-[2-[[3-methoxy-4-[3-[4-(tetrahydropyran-4-ylamino)-1-(2,2,2-trifluoroethyl)indol-2-yl]prop-2-ynylamino]phenyl]-sulfonylamino]ethoxy]ethoxy]ethoxy]ethyl]pentanamide: N-[2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]ethyl]-3-methoxy-4-[3-[4-(tetrahydropyran-4-ylamino)-1-(2,2,2-trifluoroethyl)indol-2-yl]prop-2-ynylamino]benzenesulfonamide (40.0 mg, 56.2 μmol, 1.0 equiv) in DCM (1 mL) To the solution was added TEA (28.4 mg, 281 μmol, 39.1 μL, 5.0 equiv.) and (2,5-dioxopyrrolidin-1-yl) 5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoate (19.1 mg, 56.2 μmol, 1.0 equiv.). The mixture was stirred at 20 °C for 1 h. The reaction mixture was concentrated under reduced pressure to give a residue, which was purified by preparative HPLC (column: Phenomenex Luna C18 200 mm × 40 mm × 10 μm; mobile phase: 35–70% ACN in water (+ formic acid modifier)) to give the title compound (11.2 mg, 21% yield) as a white solid.

[0516]

[0695] 1H NMR (400 MHz, DMSO-d6) δ 7.80 (t, J = 5.6 Hz, 1H), 7.35 - 7.24 (m, 2H), 7.19 (d, J = 1.8 Hz, 1H), 7.08 (s, 1H), 7.01 (t, J = 8.0 Hz, 1H), 6.83 (d, J = 8.4 Hz, 1H), 6.70 (d, J = 8.4 Hz, 1H), 6.40 (s, 1H), 6.36 - 6.28 (m, 2H), 6.22 (d, J = 7.8 Hz, 1H), 5.55 (d, J = 2.2 Hz, 1H), 4.93 (q, J = 9.2 Hz, 2H), 4.41 - 4.22 (m, 3H), 4.18 - 4.07 (m, 1H), 3.95 - 3.79 (m, 5H), 3.52 - 3.36 (m, 14H), 3.23 - 3.13 (m, 2H), 3.11 - 3.02 (m, 1H), 2.90 - 2.74 (m, 3H), 2.57 (d, J = 12.6 Hz, 1H), 2.05 (t, J = 7.4 Hz, 2H), 1.91 (d, J = 12.4 Hz, 2H), 1.57 - 1.39 (m, 5H), 1.70 - 1.38 (m, 2H), 1.36 - 1.20 (m, 2H).

[0696] LCMS[M+1]=938.3.

[0517] Biotechnology

[0697] Compound potency (IC 50The binding of the biotinylated small molecule probe BP1 to the Y220C TP53 mutant DBD was evaluated in a TR-FRET binding assay. Specifically, 5 microliters of a mixture consisting of 32 nM BP1 and 48 nM streptavidin-d2 (Cisbio) in 10 mM KH2PO4 pH 7.2, 150 mM NaCl, 0.01% BSA, 0.01% Tween-20, and 0.1 mM TCEP was added to a 384-well plate containing a 10-point dose-response titration of test compound in duplicate in 60 nL DMSO (0.6%, fcDMSO (v / v)). An additional 5 microliters of a mixture consisting of 10 nM Y220C TP53 DBD (E. coli-expressed His-TEV-P89-T312-FLAG; Uniprot ID P04637-1) and 0.3 nM MAb Anti FLAG M2-Tb cryptate (Perkin Elmer) in 10 mM KH2PO4 pH 7.2, 150 mM NaCl, 0.01% BSA, 0.01% Tween-20, and 0.1 mM TCEP was added to the 384-well plate. The mixture was incubated at 20°C. TR-FRET responses were monitored 4 and 24 hours after incubation of the BP1 and compound mixtures. Plates were read on an EnVision plate reader (Perkin Elmer) equipped with Ex / Em 615 / 665. The potency of the test compounds (IC 50 To determine competitive binding to TP53 Y220C protein in the presence of biotinylated probe BP1, TR-FRET ratios were normalized to the mean ratio of DMSO control wells (0% inhibition) and the mean minimum ratio obtained with 5 micromolar BP1 positive control compound (100% inhibition). The dose response of test compounds was calculated as IC 50 and nHill slopes were fitted using nonlinear regression with a four parameter fit. The results are shown in Table 15 below and are expressed as ranges as described here: A: IC 50 <0.100 μM, inhibition %>90%; B: IC 50 = 0.100-1.00 μM, inhibition % = 70-90%; C: IC 50= 1.00-10.0 μM, inhibition % = 50-70%; D: IC 50 = 10.0-60.0 μM, inhibition % = 10-50%; E: IC 50 >60.0μM, inhibition%<10%.

[0518]

[0698] The TR-FRET assay described above can be used to evaluate the binding affinity of both reversible and covalent TP53 Y220C ligands, stabilizers, and correctors, and can also distinguish between reversible and covalent TP53 Y220C ligands, stabilizers, and correctors. To validate the TR-FRET assay described above, we characterized a selection of TP53 Y220C ligands, stabilizers, and correctors previously described in the scientific literature. PK9301 and PK9323 are compounds described by Joerger et al. (ACS Chem. Biology 2020, 15, 657-668), which have been shown by X-ray crystallography to bind to TP53 Y220C and stabilize the mutant protein as determined by differential scanning fluorometry (DSF), and each exhibited an IC of 1.0 or higher in the TR-FRET binding assay described above. 50 PK5196 is a compound described by Jorger, Boeckler et al. (J. Am. Chem. Soc. 2012, 134, 6810-6818) that has been shown by X-ray crystallography to bind to TP53 Y220C and stabilize the mutant protein as determined by differential scanning fluorometry (DSF), nuclear magnetic resonance (NMR), and isothermal calorimetry (ITC), and has an IC of 1.5 μM and 4.1 μM in the TR-FRET assay described above. 50 = 9.7 μM. PK9301, PK9323, and PK5196 are all reversible TP53 Y220C ligands / stabilizers and therefore exhibit time-independent activity in the TR-FRET assay described above. Identical or nearly identical IC 50Values ​​are observed at both 4 and 24 hours. Generally, covalent ligands / modifiers have time-dependent activity because the covalent modification of a protein produces a sustained, cumulative effect as the reaction with the target protein progresses. Compounds of Formula I often exhibit time-dependent displacement of the TR-FRET binding probe BP1 from TP53 Y220C, distinguishing them from reversible TP53 Y220C ligands / stabilizers / correctors. In certain instances, we believe that covalent modification of mutant TP53 Y220C leads to more sustained stabilization of the mutant protein, resulting in more robust restoration of wild-type TP53 function compared to reversible ligands. This covalent mechanism of action may be advantageous for small molecule therapeutics targeted to pathologies, such as cancer, associated with or resulting from the TP53 Y220C mutant protein.

[0519] [Table 15-1]

[0520] [Table 15-2]

[0521] [Table 15-3]

[0522]

[0001] While numerous embodiments have been described, it will be apparent that the basic examples may be modified to provide other embodiments which utilize the compounds and methods of this invention. It will therefore be appreciated that the scope of this invention will be defined by the appended claims rather than by the specific embodiments that have been represented by way of example.

[0523]

[0002] The contents of all references cited throughout this application (including literature references, published patents, published patent applications, and co-pending patent applications) are expressly incorporated herein by reference in their entirety. Unless otherwise defined, all technical and scientific terms used herein are to be given the meaning commonly known to those of ordinary skill in the art.

Claims

1. Formula I: 【Chemistry 1】 or a pharmaceutically acceptable salt thereof. [In the formula, R １ is selected from optionally substituted alkyl; R ２ is hydrogen, halo, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted heterocyclyl, —NR ａ R ｂ , -OR ｃ , -NHC(O)R ｃ , —C(O)NR ｄ R ｅ , -C(O)R ｆ and -SR ｇ is selected from R ３ is hydrogen, halo, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted heterocyclyl, —NR ａ１ R ｂ１ , -OR ｃ１ , -NHC(O)R ｃ１ , —C(O)NR ｄ１ R ｅ１ , -C(O)R ｆ１ and -SR ｇ１ is selected from R ４ is halo, cyano, optionally substituted alkyl and optionally substituted alkoxy; X is halo, -S(O) ２ alkyl and —S(O)alkyl; R ａ , R ａ１ , R ｂ , R ｂ１ , R ｃ and R ｃ１ are each independently selected from hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted aryl, and optionally substituted heterocyclyl; R ｄ , R ｅ and R ｇ are each independently hydrogen, optionally substituted alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted heterocyclyl, —C(O)R′, —C(O)OR′, —C(O)NR′R″, —S(O)R′, and —S(O) ２ R'; or R ｄ and R ｅ together with the nitrogen atom to which they are attached form an optionally substituted heterocyclyl or an optionally substituted heteroaryl; R ｄ１ , R ｅ１ and R ｇ１ are each independently hydrogen, optionally substituted alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted heterocyclyl, —C(O)R′ １ , —C(O)OR′ １ , —C(O)NR′ １ R'' １ , —S(O)R′ １ and -S(O) ２ R' １ or R ｄ１ and R ｅ１ together with the nitrogen atom to which they are attached form an optionally substituted heterocyclyl or an optionally substituted heteroaryl; R ｆ and R ｆ１ are each independently selected from hydrogen, optionally substituted alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, and optionally substituted heterocyclyl; R', R' １ , R″ and R″ １ are each independently hydrogen and optionally substituted (C １ ~C ４ ) alkyl; p is 0, 1 or 2.

2. 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein p is 0.

3. X is chloro, bromo, fluoro, or —SO ２ CH ３ and -SOCH ３ 3. The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, selected from:

4. X is chloro, bromo, fluoro and —SO ２ CH ３ 4. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, selected from:

5. 5. The compound of any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, wherein X is chloro.

6. R １ is (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, cyano (C １ ~C ４ ) alkyl and —(C １ ~C ４ ) alkyl(C ３ ~C ６ 6. The compound of any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein:

7. R １ is (C １ ~C ４ ) alkyl and halo(C １ ~C ４ 7. The compound of any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, wherein:

8. R １ is -CH ２ CH ３ , -CH ２ CF ３ , -CH ２ CF ２ CH ３ , -CH ２ CF ２ CF ３ , -CH ２ CN and -CH ２ 7. The compound of any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, selected from: -cyclopropyl.

9. R １ is -CH ２ CF ３ 9. The compound according to any one of claims 1 to 8, wherein:

10. R ２ is hydrogen, halo, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, (C ３ ~C ６ ) cycloalkyl, -(C １ ~C ４ ) alkyl(C ３ ~C ６ ) cycloalkyl, 5- to 7-membered heteroaryl, —(C １ ~C ４ ) alkyl [5- to 7-membered heteroaryl], phenyl, -(C １ ~C ４ ) alkylphenyl, 4- to 6-membered heterocyclyl, -(C １ ~C ４ ) alkyl [4- to 6-membered heterocyclyl], —NR ａ R ｂ , -OR ｃ , -(C １ ~C ４ ) alkyl OR ｃ , -C(O)R ｆ , —C(O)NR ｄ R ｅ , -(C １ ~C ４ ) alkylNR ｄ R ｅ , -(C １ ~C ４ ) alkylNR ａ C(O)R ｂ , -(C １ ~C ４ ) alkylC(O)R ｆ , -(C １ ~C ４ ) alkylC(O)NR ｄ R ｅ and -SR ｇ is selected from the group consisting of: ３ ~C ６ ) For each occurrence of cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclyl, said groups each may be selected from the group R ５ and optionally substituted with 1 to 3 groups selected from R ａ , R ｂ and R ｃ are each independently hydrogen, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, (C ３ ~C ６ ) cycloalkyl, -(C １ ~C ４ ) alkyl(C ３ ~C ６ ) cycloalkyl, 5- to 7-membered heteroaryl, —(C １ ~C ４ ) alkyl [5- to 7-membered heteroaryl], phenyl, —(C １ ~C ４ ) alkylphenyl, 4- to 6-membered heterocyclyl, -(C １ ~C ４ ) alkyl [4- to 6-membered heterocyclyl], wherein (C ３ ~C ６ ) For each occurrence of cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclyl, said groups each may be selected from the group R ６ and optionally substituted with 1 to 3 groups selected from R ｄ , R ｅ and R ｇ are each independently hydrogen, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, -(C １ ~C ４ ) alkyl OR', -(C １ ~C ４ ) alkylNR'R'', -(C １ ~C ４ ) alkylC(O)NR′R″, —(C １ ~C ４ ) alkylC(O)R', -(C １ ~C ４ ) alkylC(O)OR', -(C １ ~C ４ ) alkylS(O)R', -(C １ ~C ４ ) alkylS(O) ２ R', (C １ ~C ４ ) alkylphenyl, phenyl, (C ３ ~C ６ ) cycloalkyl, -(C １ ~C ４ ) alkyl [5- to 7-membered heteroaryl], 5- to 7-membered heteroaryl, 4- to 6-membered heterocyclyl, -(C １ ~C ４ ) alkyl [4- to 6-membered heterocyclyl], —C(O)R′, —C(O)OR′, —C(O)NR′R″, —S(O)R′ and —S(O) ２ R′, where (C ３ ~C ６ ) For each occurrence of cycloalkyl, phenyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclyl, said groups each may be selected from the group R ７ or R ｄ and R ｅ together with the nitrogen atom to which they are attached, R ７ forming a 4- to 6-membered heterocyclyl or a 5- to 7-membered heteroaryl, each optionally substituted with 1 to 3 groups selected from R ｆ is hydrogen, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, phenyl, (C ３ ~C ６ ) cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclyl, wherein said phenyl, (C ３ ~C ６ ) Cycloalkyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclyl are each R ８ and optionally substituted with 1 to 3 groups selected from R ａ , R ５ , R ６ , R ７ and R ８ are each independently a halo, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkoxy, halo (C １ ~C ４ ) alkoxy, (C ３ ~C ６ ) cycloalkyl, —OR′, —NR′R″, —(C １ ~C ４ ) alkylNR'R''-(C １ ~C ４ ) alkylC(O)NR'R'', oxo, -(C １ ~C ４ ) alkylOR', -C(O)R', -S(O)R' and -S(O) ２ R' is selected from R′ and R″ are each independently hydrogen, (C １ ~C ４ ) alkyl and (C ３ ~C ６ 10. The compound of any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, wherein:

11. R ２ is hydrogen, halo, (C １ ~C ４ ) alkyl, (C ３ ~C ６ ) cycloalkyl, 5- to 7-membered heteroaryl, phenyl, 4- to 6-membered heterocyclyl, —NR ａ R ｂ , -OR ｃ , —C(O)NR ｄ R ｅ , -(C １ ~C ４ ) alkylNR ｄ R ｅ , -(C １ ~C ４ ) alkylNR ａ C(O)R ｂ and -(C １ ~C ４ ) alkylC(O)NR ｄ R ｅ wherein 5- to 7-membered heteroaryl, phenyl, (C ３ ~C ６ ) For each occurrence of cycloalkyl and 4- to 6-membered heterocyclyl, said groups each may be selected from R ５ 11. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

12. R ２ is hydrogen, halo, (C １ ~C ４ ) alkyl, cyclopropyl, pyrazolyl, phenyl, pyrrolidinyl, piperazinyl, piperidinyl, -NR ａ R ｂ , -OR ｃ , —C(O)NR ｄ R ｅ , -(C １ ~C ４ ) alkylNR ｄ R ｅ , -(C １ ~C ４ ) alkylNR ａ C(O)R ｂ and -(C １ ~C ４ ) alkylC(O)NR ｄ R ｅ wherein said cyclopropyl, pyrazolyl, phenyl, pyrrolidinyl, piperazinyl and piperidinyl are each selected from R ５ 12. The compound of any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

13. R ａ , R ｂ and R ｃ are each independently hydrogen, (C １ ~C ４ ) alkyl, -(C １ ~C ４ ) alkyl [5- to 7-membered heteroaryl], (C ３ ~C ６ ) cycloalkyl, phenyl, -(C １ ~C ４ ) alkylphenyl, 4- to 6-membered heterocyclyl and -(C １ ~C ４ ) alkyl[4- to 6-membered heterocyclyl], wherein for each occurrence of 5- to 7-membered heteroaryl, 4- to 6-membered heterocyclyl, and phenyl, said groups are each selected from R ６ 11. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

14. R ５ and R ６ are each independently a halo, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, -(C １ ~C ４ ) alkyl OR' and S(O) ２ R' is selected from the group consisting of: R', ...

15. R ２ is hydrogen, chloro, methyl, cyclopropyl, —OCH ２ CF ３ , -CH ２ NHC(O)CH ３ , -CH ２ NHSO ２ CH ３ , -CH ２ C(O)NHCH ３ , -CH ２ C(O)N(CH ３ ) ２ , -CH ２ C(O)NHCH ２ CF ３ , -CH ２ C(O)NHCH ２ OCH ３ , -CH ２ C(O)NH ２ , -CH ２ C(O)N(CH ３ ) (CH ２ CF ３ ), -CH ２ C(O)NH(CH ２ ) ２ SO ２ CH ３ , -C(O)N(CH ３ ) ２ , -C(O)NHCH ３ , 【Chemistry 2】 15. The compound of any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, selected from:

16. R ２ is hydrogen, chloro and -OCH ２ CF ３ 16. The compound of any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof, selected from:

17. R ２ 17. The compound of any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, wherein is hydrogen.

18. R ３ Halo, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, (C ３ ~C ６ ) cycloalkyl, -(C １ ~C ４ ) alkyl(C ３ ~C ６ ) cycloalkyl, 5- to 7-membered heteroaryl, —(C １ ~C ４ ) alkyl [5- to 7-membered heteroaryl], phenyl, —(C １ ~C ４ ) alkyl OR ｃ１ , -(C １ ~C ４ ) alkylphenyl, 4- to 10-membered heterocyclyl, -(C １ ~C ４ ) alkyl [4- to 10-membered heterocyclyl], —NR ａ１ R ｂ１ , -NHC(O)R ｃ１ , -OR ｃ１ , -(C １ ~C ４ ) alkyl OR ｃ１ , -C(O)R ｆ１ , —C(O)NR ｄ１ R ｅ１ , -(C １ ~C ４ ) alkylNR ｇ１ R ｈ１ , -(C １ ~C ４ ) alkylC(O)R ｆ１ , -(C １ ~C ４ ) alkylC(O)NR ｄ１ R ｅ１ and -SR ｇ１ is selected from the group consisting of: ３ ~C ６ ) For each occurrence of cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 10-membered heterocyclyl, said groups each may be selected from the group R ５ａ and optionally substituted with 1 to 3 groups selected from R ａ１ , R ｂ１ and R ｃ１ are each independently hydrogen, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, (C ３ ~C ６ ) cycloalkyl, -(C １ ~C ４ ) alkyl(C ３ ~C ６ ) cycloalkyl, 5- to 7-membered heteroaryl, —(C １ ~C ４ ) alkyl [5- to 7-membered heteroaryl], phenyl, —(C １ ~C ４ ) alkylphenyl, 4- to 6-membered heterocyclyl, -(C １ ~C ４ ) alkyl [4- to 6-membered heterocyclyl], wherein (C ３ ~C ６ ) For each occurrence of cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclyl, said groups each may be selected from the group R ６ａ and optionally substituted with 1 to 3 groups selected from R ｄ１ , R ｅ１ , R ｇ１ and R ｈ１ are each independently hydrogen, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, -(C １ ~C ４ ) alkyl OR' １ , -(C １ ~C ４ ) alkylNR' １ R'' １ , -(C １ ~C ４ ) alkylC(O)NR' １ R'' １ , -(C １ ~C ４ ) alkylC(O)R' １ , -(C １ ~C ４ ) alkylC(O)OR' １ , -(C １ ~C ４ ) alkylS(O)R' １ , -(C １ ~C ４ ) alkylS(O) ２ R' １ , (C １ ~C ４ ) alkylphenyl, phenyl, (C ３ ~C ６ ) cycloalkyl, -(C １ ~C ４ ) alkyl [5- to 7-membered heteroaryl], 5- to 7-membered heteroaryl, 4- to 8-membered heterocyclyl, -(C １ ~C ４ ) alkyl [4- to 8-membered heterocyclyl], —C(O)R′ １ , —C(O)OR′ １ , —C(O)NR′ １ R'' １ , —S(O)R′ １ and -S(O) ２ R' １ is selected from the group consisting of: ３ ~C ６ ) For each occurrence of cycloalkyl, phenyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclyl, said groups each may be selected from the group R ７ａ or R ｄ１ and R ｅ１ together with the nitrogen atom to which they are attached, R ７ａ forming a 4- to 8-membered heterocyclyl or a 5- to 7-membered heteroaryl, each optionally substituted with 1 to 3 groups selected from R ｆ１ is hydrogen, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, phenyl, (C ３ ~C ６ ) cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclyl, wherein said phenyl, (C ３ ~C ６ ) Cycloalkyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclyl are each R ８ａ and optionally substituted with 1 to 3 groups selected from R ５ａ , R ６ａ , R ７ａ and R ８ａ are each independently halo, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkoxy, halo (C １ ~C ４ ) alkoxy, (C ３ ~C ６ ) cycloalkyl, 5- to 7-membered heteroaryl, cyano, —NR′ １ R'' １ , -(C １ ~C ４ ) alkylNR' １ R'' １ , -(C １ ~C ４ ) alkylC(O)OR' １ , -(C １ ~C ４ ) alkylC(O)NR' １ R'' １ , oxo, -(C １ ~C ４ ) alkyl OR' １ , —C(O)NR′ １ R'' １ , -OR' １ , C(O)OR' １ , —C(O)R′ １ , —S(O)R′ １ , -S(O) ２ R' １ , R' １ and 4- to 6-membered heterocyclyl optionally substituted with 1 to 3 groups selected from R' １ and R'' １ are each independently hydrogen, (C １ ~C ４ ) alkyl, (C ３ ~C ６ 18. The compound of any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, wherein R is selected from cycloalkyl and 4- to 7-membered heterocyclyl.

19. R ３ is -(C １ ~C ４ ) alkyl [4- to 10-membered heterocyclyl], -(C １ ~C ４ ) alkyl [5- to 7-membered heteroaryl], 4- to 10-membered heterocyclyl, -(C １ ~C ４ ) alkyl OR ｃ１ , -NR ａ１ R ｂ１ , -NHC(O)R ｃ１ , -C(O)R ｆ１ , —C(O)NR ｄ１ R ｅ１ and -(C １ ~C ４ ) alkylNR ｇ１ R ｈ１ wherein (C ３ ~C ６ ) Cycloalkyl, 5- to 7-membered heteroaryl, 4- to 10-membered heterocyclyl, and 4- to 9-membered heterocyclyl are each R ５ａ 19. The compound of any one of claims 1 to 18, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

20. R ３ is -(C １ ~C ４ ) alkyl OR ｃ１ , 4- to 10-membered heterocyclyl, -(C １ ~C ４ ) alkyl [4- to 10-membered heterocyclyl], —NR ａ１ R ｂ１ , -NHC(O)R ｃ１ , -C(O)R ｆ１ , —C(O)NR ｄ１ R ｅ１ , -(C １ ~C ４ ) alkylNR ｇ１ R ｈ１ wherein each 4- to 10-membered heterocyclyl is selected from R ５ａ 20. The compound of any one of claims 1 to 19, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

21. R ３ is -(C １ ~C ４ ) alkyl OR ｃ１ , -NR ａ１ R ｂ１ , -NHC(O)R ｃ１ , -C(O)R ｆ１ , —C(O)NR ｄ１ R ｅ１ , -(C １ ~C ４ ) alkylNR ｇ１ R ｈ１ , -(C １ ~C ４ ) alkyl[piperazinyl], -(C １ ~C ４ ) alkyl[piperidinyl], -(C １ ~C ４ ) alkyl [morpholinyl], -(C １ ~C ４ ) alkyl[pyrrolidinyl], -(C １ ~C ４ ) alkyl [diazepanyl], -(C １ ~C ４ ) alkyl[azetidinyl], piperazinyl, and tetrahydropyridinyl, wherein said piperidinyl, morpholinyl, pyrrolidinyl, diazepanyl, tetrahydropyridinyl, azetidinyl, and each occurrence of piperazinyl is selected from R ５ａ 21. The compound of any one of claims 1 to 20, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

22. R ５ａ is (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, (C ３ ~C ６ ) cycloalkyl, 5- to 7-membered heteroaryl, —NR′ １ R'' １ , -(C １ ~C ４ ) alkylNR' １ R'' １ , -(C １ ~C ４ ) alkylC(O)OR' １ , oxo, -(C １ ~C ４ ) alkyl OR' １ , —C(O)NR′ １ R'' １ , -OR' １ , C(O)OR' １ , —C(O)R′ １ , and R' １ 22. The compound of any one of claims 1 to 21, or a pharmaceutically acceptable salt thereof, wherein R is selected from 4- to 6-membered heterocyclyl optionally substituted with 1 to 3 groups selected from:

23. R ５ａ is (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, cyclopropyl, imidazolyl, piperazinyl, pyrazolyl, triazolyl, terazolyl, azetidinyl, oxo, -N[(C １ ~C ４ ) alkyl] ２ , -(C １ ~C ４ ) alkyl NH ２ , -(C １ ~C ４ ) alkylNH[(C １ ~C ４ ) alkyl], -(C １ ~C ４ ) alkylN[(C １ ~C ４ ) alkyl] ２ , -(C １ ~C ４ ) alkylC(O)N[(C １ ~C ４ ) alkyl] ２ , -(C １ ~C ４ ) alkylOH, —(C １ ~C ４ ) alkylO(C １ ~C ４ ) alkyl, -(C １ ~C ４ ) alkyl C(O)OH, C(O)OH, —OH, C(O)NH ２ , and —C(O)(C １ ~C ４ 23. The compound of any one of claims 1 to 22, or a pharmaceutically acceptable salt thereof, wherein:

24. R ｃ１ is selected from 5- to 7-membered heteroaryl and 4- to 6-membered heterocyclyl, where the 5- to 7-membered heteroaryl and the 4- to 6-membered heterocyclyl are each selected from R ６ａ 24. The compound of any one of claims 1 to 23, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

25. R ｃ１ are respectively R ６ａ 25. The compound of any one of claims 1 to 24, or a pharmaceutically acceptable salt thereof, which is piperidinyl or pyridinyl optionally substituted with one to three groups selected from:

26. R ６ａ is (C １ ~C ４ 26. The compound of any one of claims 1 to 25, or a pharmaceutically acceptable salt thereof, wherein:

27. R ａ１ and R ｂ１ are each independently hydrogen, -(C １ ~C ４ ) alkyl [5- to 7-membered heteroaryl] and 4- to 6-membered heterocyclyl, wherein at each occurrence of 5- to 7-membered heteroaryl and 4- to 6-membered heterocyclyl, said groups are each selected from R ６ａ 27. The compound of any one of claims 1 to 26, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

28. R ａ１ and R ｂ１ are each independently hydrogen and -(C １ ~C ４ ) alkyl [5- to 7-membered heteroaryl], wherein said 5- to 7-membered heteroaryl is selected from R ６ａ 28. The compound of any one of claims 1 to 27, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

29. R ａ１ is hydrogen, and R ｂ１ is -(C １ ~C ４ ) alkyl[pyridinyl], wherein said pyridinyl is R ６ａ 29. The compound of any one of claims 1 to 28, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

30. R ｆ１ is R ８ａ 30. The compound of any one of claims 1 to 29, or a pharmaceutically acceptable salt thereof, which is a 4- to 6-membered heterocyclyl optionally substituted with 1 to 3 groups selected from:

31. R ｆ１ is R ８ａ 31. The compound of any one of claims 1 to 30, or a pharmaceutically acceptable salt thereof, which is piperazinyl optionally substituted with 1 to 3 groups selected from:

32. R ８ａ is (C １ ~C ４ 32. The compound of any one of claims 1 to 31, or a pharmaceutically acceptable salt thereof, wherein:

33. R ｄ１ and R ｅ１ are each independently hydrogen, -(C １ ~C ４ ) alkylNR' １ R'' １ , (C １ ~C ４ ) alkyl [5- to 7-membered heteroaryl] and 4- to 8-membered heterocyclyl, wherein said 5- to 7-membered heteroaryl and 4- to 8-membered heterocyclyl are each selected from R ７ａ 33. The compound of any one of claims 1 to 32, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

34. R ｄ１ and R ｅ１ are each independently hydrogen, -(C １ ~C ４ ) alkylN[(C １ ~C ４ ) alkyl] ２ , (C １ ~C ４ ) alkyl[pyridinyl] and piperidinyl, wherein said pyridinyl and piperidinyl are each selected from R ７ａ 34. The compound of any one of claims 1 to 33, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

35. R ｇ１ and R ｈ１ are each independently hydrogen, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, -(C １ ~C ４ ) alkyl OR' １ , -(C １ ~C ４ ) alkylNR' １ R'' １ , -(C １ ~C ４ ) alkylC(O)NR' １ R'' １ , -(C １ ~C ４ ) alkylC(O)R' １ , (C １ ~C ４ ) alkylphenyl, —(C １ ~C ４ ) alkyl [5- to 7-membered heteroaryl] and 4- to 8-membered heterocyclyl, wherein said phenyl, 5- to 7-membered heteroaryl and 4- to 8-membered heterocyclyl are each selected from R ７ａ 35. The compound of any one of claims 1 to 34, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

36. R ｇ１ and R ｈ１ are each independently hydrogen, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, -(C １ ~C ４ ) alkyl[pyridinyl], -(C １ ~C ４ ) alkylO(C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkylphenyl, —(C １ ~C ４ ) alkylN[(C １ ~C ４ ) alkyl] ２ , pyridinyl, piperidinyl, pyrrolidinyl, -(C １ ~C ４ ) alkylC(O)N[(C １ ~C ４ ) alkyl] ２ , Halo (C １ ~C ４ ) alkyl, -(C １ ~C ４ ) alkylC(O)(morpholinyl), wherein said phenyl, pyridinyl, piperidinyl, pyrrolidinyl and morpholinyl are each selected from R ７ａ 36. The compound of any one of claims 1 to 35, or a pharmaceutically acceptable salt thereof, optionally substituted with 1 to 3 groups selected from:

37. R ７ａ is (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, halo, (C １ ~C ４ 37. The compound of any one of claims 1 to 36, or a pharmaceutically acceptable salt thereof, wherein: R is selected from alkoxy and cyano.

38. R ３ は、-CH ２ NHCH ３ 、-CH ２ N(CH) ３ ) ２ 、-(CH ２ ) ２ N(CH) ３ ) ２ 、-CH ２ N(CH) ３ (CH) ２ ) ２ OCH ３ 、-CH ２ N(CH) ３ (CH) ２ ) ２ N(CH) ３ ) ２ 、-CH ２ NH(CH) ２ CH ３ )、-CH ２ N(CH) ３ (CH) ２ ) ３ N(CH) ３ ) ２ 、-CH ２ NH(CH) ２ ) ２ N(CH) ２ CH ３ ) ２ CH ２ NHC(CH) ３ ) ３ 、-CH ２ N(CH) ３ )CH ２ C(O)N(CH) ３ ) ２ 、-C(O)NHCH ２ N(CH) ３ ) ２ 、 【Chemistry 3-1】 【Chemistry 3-2】 【Chemistry 3-3】 [Chemistry 3-4] [Transformation 3-5] 38. The compound of any one of claims 1 to 37, or a pharmaceutically acceptable salt thereof, selected from: 【Request Item 39】 【Chemistry 4-1】 【Chemistry 4-2】 【Chemistry 4-3】 【Chemistry 4-4】 [Chemistry 4-5] [Chemistry 4-6] 【Chemistry 4-7】 【Chemistry 4-8】 【Chemistry 4-9】 【Chemistry 4-10】 【Chemistry 4-11】 【Chemistry 4-12】 【Chemistry 4-13】 【Chemistry 4-14】 【Chemistry 4-15】 【Chemistry 4-16】 【Chemistry 4-17】 【Chemistry 4-18】 【Chemistry 4-19】 【Chemistry 4-20】 【Chemistry 4-21】 【Chemistry 4-22】 【Chemistry 4-23】 【Chemistry 4-24】 【Chemistry 4-25】 【Chemistry 4-26】 【Chemistry 4-27】 【Chemistry 4-28】 [Chemistry 4-29] 【Chemistry 4-30】 【Chemistry 4-31】 【Chemistry 4-32】 【Chemistry 4-33】 Selected from:

2. The compound of claim 1 or a pharmaceutically acceptable salt thereof.

40. 40. A compound according to any one of claims 1 to 39, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

41. 41. A method of treating a condition responsive to activation of wild-type tumor suppressor protein TP53 function in a patient in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 39, or a composition according to claim 40.

42. 42. The method of claim 41, wherein the condition is cancer.

43. 43. The method of claim 41 or 42, wherein the condition is a cancer harboring a Y220C mutation.

44. 44. The method of any one of claims 41 to 43, wherein the cancer is a solid tumor or a hemolytic malignancy.

45. 45. The method of any one of claims 41 to 44, wherein the cancer is selected from lung cancer, ovarian cancer, breast cancer, colorectal cancer, pancreatic cancer, glioma, glioblastoma, endometrial cancer, esophageal cancer, gastric cancer, prostate cancer, head and neck cancer, bladder cancer, acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), sarcoma, and melanoma.

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