Compounds and compositions targeting tp53-y220c mutants

Abstract

Provided are compounds of Formula (I) (I); and pharmaceutically acceptable salts and compositions thereof, which are useful in the treatment of various conditions associated with activation of p53.

Description

Related applications

[0001] This application claims priority to U.S. Provisional Application No. 63 / 619,000, filed January 9, 2024, the entire contents of which are incorporated herein by reference. Background Technology

[0002] The transcription factor TP53 acts as a tumor suppressor gene and is inactivated by mutations in approximately 50% of all tumors. TP53 regulates multiple intracellular metabolic pathways, including DNA damage repair, apoptosis, and senescence. The Y220C mutation is a common TP53 missense mutant, associated with over 100,000 new cancer cases globally each year, primarily breast and ovarian cancer. The Y220C mutation results in a significant structural change in the TP53 protein and is known to form a novel protein cavity estimated to accommodate small molecule drug candidates. Crucially, the mutation-induced cleft is located away from the TP53 surface involved in DNA recognition or protein-protein interactions, enabling the development of targeted chemotherapeutic agents that stabilize the DNA-binding domain without interfering with its natural substrate binding. Stabilization can restore TP53 function, thereby reactivating the TP53 tumor suppressor gene pathway and shrinking or killing tumors. Summary of the Invention

[0003] This article provides compounds having formula I: (I); and pharmaceutically acceptable salts and compositions thereof, wherein R 1 R 2 R 3 A, X, Y, and Z are as described herein. In one aspect, the disclosed Formula I compound and its pharmaceutically acceptable salt are used as covalent modifiers of TP53 Y220C. In one aspect, the disclosed Formula I compound and its pharmaceutically acceptable salt stabilize TP53 Y220C. In one aspect, the disclosed Formula I compound and its pharmaceutically acceptable salt restore the function of the wild-type tumor suppressor protein TP53 (WT TP53). In one aspect, the compound described is believed to be a covalent modifier of TP53 Y220C, which causes stabilization of the mutant protein and can restore the wild-type function of the dysfunctional mutant protein. See, for example, the Examples section below.

[0004] It also includes pharmaceutical compositions comprising the disclosed Formula I compound and its pharmaceutically acceptable salts, and methods for preparing the same. Detailed Implementation

[0005] 1. General description of compounds In the first embodiment, a compound of formula I is provided herein: (I); R 1 Selected from alkyl groups with optional substitution; R 2 Selected from hydrogen, halogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted heterocyclic, -NR a R b -OR c -NHC(O)R c -C(O)NR d R e -C(O)R f and -SR g ; R 3 Selected from hydrogen, halogen, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted heterocyclic, -NR a1 R b1 -OR c1 -NHC(O)R c1 -C(O)NR d1 R e1 -C(O)R f1 and -SR g1 ; X is selected from halogens, -S(O)2 alkyl groups, and –S(O) alkyl groups; The variables that are N if one of A, Y, or Z is not N are -CH-; R a R a1 R b R b1 R c and R c1 Each is independently selected from hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted aryl and optionally substituted heterocyclic groups; R d and R e Each is independently selected from hydrogen, optionally substituted alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted heterocyclic, -C(O)R', -C(O)OR', -C(O)NR'R'', -S(O)R' and -S(O)2R'; or R d and R e Together with the nitrogen atom to which it is attached, it forms an optionally substituted heterocyclic group or an optionally substituted heteroaryl group; R d1 and R e1Each is independently selected from hydrogen, optionally substituted alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted heterocyclic, -C(O)R' 1 -C(O)OR' 1 -C(O)NR' 1 R'' 1 -S(O)R' 1 and -S(O)2R' 1 Or R d1 and R e1 Together with the nitrogen atom to which it is attached, it forms an optionally substituted heterocyclic group or an optionally substituted heteroaryl group; R f and R f1 Each is independently selected from hydrogen, optionally substituted alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, and optionally substituted heterocyclic; and R'、R' 1 , R'' and R'' 1 Each is independently selected from hydrogen and optionally substituted (C1-C4) alkyl groups.

[0006] 2. definition When used to describe chemical groups that may have multiple connection points, the hyphen (-) indicates the connection point between the group and the variable it defines. For example, -NR a R b This indicates that the bonding point of the group is located on a nitrogen atom.

[0007] The terms “halogen” and “halogen” refer to atoms selected from fluorine (fluorinated, -F), chlorine (chlorinated, -Cl), bromine (brominated, -Br), and iodine (iodinated, -I).

[0008] When the term "alkyl" is used alone or as part of a larger part, such as "halogenated alkyl", it refers to a saturated straight-chain or branched monovalent hydrocarbon group.

[0009] "Alkoxy" refers to an alkyl group linked by an oxygen atom and is represented by -O-alkyl. For example, "(C1-C4)alkoxy" includes methoxy, ethoxy, propoxy, and butoxy.

[0010] The term "haloalkyl" includes monohaloalkyl, polyhaloalkyl, and perhaloalkyl, wherein the halogen is independently selected from fluorine, chlorine, bromine, and iodine.

[0011] "Haloalkoxy" is a haloalkyl group connected to another part via an oxygen atom, such as -OCHF2 or -OCF3.

[0012] The term "oxo" indicates that the =O group is present.

[0013] The term "heteroaryl," used alone or as part of a larger part, refers to a 5- to 12-membered aromatic group containing 1 to 4 heteroatoms selected from N, O, and S. Heteroaryls can be monocyclic or bicyclic. Monocyclic heteroaryls include, for example, thienyl, furanyl, pyrroleyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, triazinyl, tetraazinyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrazinyl, etc. Bicyclic heteroaryl compounds include groups in which a monocyclic heteroaryl ring is fused with one or more aryl or heteroaryl rings. Non-limiting examples include indolyl, imidazopyridyl, benzoxazolyl, benzoxodiazolyl, indazole, benzimidazolyl, benzothiazolyl, quinolinyl, quinazolinyl, quinoxalinyl, pyrrolopyridyl, pyrrolopyrimidyl, pyrazolopyridyl, thienopyridyl, thienopyrimidyl, indolazinyl, purinyl, naphridyl, and pteridinyl. It should be understood that, when specifically specified, optional substituents on the heteroaryl group may be present at any substituted position, including, for example, the position to which the heteroaryl group is attached.

[0014] The term "heterocyclic group" refers to a 5- to 12-membered saturated or partially unsaturated heterocycle containing 1 to 4 heteroatoms independently selected from N, O, and S. It can be monocyclic, bicyclic (e.g., bridged, fused, or spirobicyclic), or tricyclic. The heterocyclic ring can be attached to any heteroatom or carbon atom of its side group, thereby forming a stable structure. Examples of such saturated or partially unsaturated heterocyclic groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, pyrrolylalkyl, pyridoneyl, pyrrolidoneyl, piperidinyl, oxazolyl, piperazineyl, dioxalyl, dioxopentyl, morpholinyl, dihydrofuranyl, dihydropyranyl, dihydropyridinyl, tetrahydropyridinyl, dihydropyrimidinyl, oxoheterocyclic butyl, azoheterocyclic butyl, and tetrahydropyrimidinyl. Heterocyclic groups can be monocyclic or bicyclic. The term "heterocyclic group" also includes, for example, an unsaturated heterocyclic group fused with another unsaturated heterocyclic group or aryl or heteroaromatic ring, such as, for example, tetrahydronaphthidine, indolinone, dihydropyrrolotriazole, imidazopyrimidine, quinolinone, and dioxaspirodecane. It should also be understood that, when specifically specified, optional substituents on the heterocyclic group may be present at any substituted position, including, for example, positions where the heterocyclic group is attached (e.g., in the case of an optionally substituted heterocyclic group or an optionally substituted heterocyclic group).

[0015] The term "spiral" refers to two rings sharing a single ring atom (e.g., carbon).

[0016] The term "fusion" refers to two rings sharing two adjacent ring atoms.

[0017] The term "bridging" refers to two rings sharing three ring atoms.

[0018] The term "cycloalkyl," used alone or as part of a larger part, refers to a saturated cyclic aliphatic monocyclic or bicyclic system as described herein, having 3 to 10 carbon ring atoms unless otherwise specified. Monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, and cyclooctyl. It is understood that, when specified, optional substituents on the cycloalkyl or cycloaliphatic group may be present at any substituted position, including, for example, the position to which the cycloalkyl group is attached.

[0019] The term "optionally substituted" means that one or more hydrogen atoms in the specified moiety can be replaced by suitable substituents. Unless otherwise stated, the "optionally substituted" group may have suitable substituents at each substituted position of the group, provided that the valence allows. Optional substituents include, but are not limited to, one or more groups selected from: cyano (–CN), halogen, 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 Riii –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 Riii、–NR i S(O)2NR ii R iv –SR i –S(O)R i –S(O)2R i –S(O)NR ii R iv –S(O)2NR ii R iv Alkyl, heteroalkyl, alkenyl, ynyl, cycloalkyl, aryl, heteroaryl, and heterocyclic groups, wherein each of the alkyl, heteroalkyl, alkenyl, ynyl, cycloalkyl, aryl, heteroaryl, and heterocyclic groups is further optionally substituented by one or more (in one embodiment, one, two, three, or four) substituents Q. a Replace, where each R i R ii R iii and R iv Independently alkyl, heteroalkyl, alkenyl, ynyl, cycloalkyl, aryl, heteroaryl, or heterocyclic, each optionally substituent Q by one or more (in one embodiment, one, two, three, or four) substituents. a Replace, or R ii and R iii Together with the N atoms they are attached to, they form Q groups that are optionally substituented by one or more (in one embodiment, one, two, three, or four) substituents. aSubstituted heterocyclic groups, where each Q a Independently selected from cyano, halogen, imino, nitro, oxo, alkyl, heteroalkyl, alkenyl, alkynyl, cycloalkyl, C 6-14 Aryl, heteroaryl, heterocyclic, –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)R v –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) Rviii –NRv 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)2NR vi R vii –SR v –S(O)R v –S(O)2R v –S(O)NR vi R vii and –S(O)2NR vi R vii ;where R v R vi R vii and R viii Each is independently alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heteroaryl, or heterocyclic; or (iii) R vi and R viii Together with the N atoms they are attached to, they form heterocyclic groups.

[0020] In some respects, when specified, one or more hydrogen atoms on the disclosed compound may be substituted with deuterium. Such deuterated compounds may have one or more improved pharmacokinetic or pharmacodynamic properties (e.g., longer half-life) compared to equivalent “non-deuterated” compounds.

[0021] One or more compounds described herein can exist in a variety of tautomeric forms and are part of this disclosure. The terms "tautomer" or "tautomeric" refer to two or more interconvertible compounds / substituents resulting from at least one form of hydrogen atom migration and at least one change in valence. All isomers of such compounds are explicitly included. Therefore, when a compound herein is represented by a structural formula or designated by a chemical name, all possible tautomeric forms of that compound are encompassed within that structural formula.

[0022] Compounds with one or more chiral centers can exist in a variety of stereoisomers. Stereoisomers are compounds that differ only in spatial arrangement. Stereoisomers include all diastereomers, enantiomers, and epimers, as well as racemates and mixtures thereof. "Geometric isomers" refer to isomers in which the substituents are oriented differently relative to the carbon-carbon double bond, cycloalkyl ring, or bridged bicyclic system. The atoms on either side of the carbon-carbon double bond (except H) can be E (substituents on opposite sides of the carbon-carbon double bond) or Z (substituents on the same side) configurations. "cis" means the substituents are on the same side of the ring, while "trans" means the substituents are on opposite sides of the ring.

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

[0024] When geometric isomers are described by name or structure, the indicated isomer has an enrichment relative to the opposite isomer greater than 50%, 60%, 70%, 80%, 90%, 99%, or 99.9%. "Enrichment of the indicated isomer relative to the opposite isomer" is a molar percentage, determined by dividing the number of compounds having the indicated geometry by the total number of all compounds in the mixture having the same or opposite geometry.

[0025] When a disclosed compound is named or described by its structure without indicating its stereochemistry, it should be understood that the name or structure covers one of the possible stereoisomers or geometric isomers without other isomers, or a mixture of the stereoisomers or geometric isomers covered.

[0026] The terms "subject" and "patient" are used interchangeably and refer to mammals in need of treatment, such as companion animals (e.g., dogs, cats, etc.), farm animals (e.g., cattle, pigs, horses, sheep, goats, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, etc.). Typically, a subject is a person in need of treatment.

[0027] The terms “inhibition,” “hindrance,” or “inhibitory” include a reduction in the baseline activity of a biological activity or process.

[0028] As used herein, the terms “treatment,” “management,” and “curing” refer to reversing, alleviating, delaying the onset of a disease or disorder or one or more symptoms thereof as described herein, or inhibiting its progression. In some respects, treatment may be administered after one or more symptoms have appeared, i.e., therapeutic treatment. In other respects, treatment may be administered even when there are no symptoms. For example, susceptible individuals may be treated before symptoms appear (e.g., based on a history of symptoms and / or based on exposure to a specific organism or other susceptibility factor), i.e., preventative treatment. Treatment may also continue after symptoms have disappeared, for example, to delay recurrence.

[0029] The term "pharmaceutically acceptable carrier" refers to a non-toxic carrier, adjuvant, or loading agent that does not impair the pharmacological activity of the compound formulated with it. Pharmaceutically acceptable carriers, adjuvants, or loading agents 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), buffering substances (such as phosphates, glycine), sorbic acid, potassium sorbate, mixtures of partial glycerides 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 carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and lanolin.

[0030] For use in pharmaceuticals, the salts of the compounds described herein refer to non-toxic, pharmaceutically acceptable salts. Pharmaceutically acceptable salt forms include pharmaceutically acceptable acidic / anionic or basic / cationic salts. Suitable pharmaceutically acceptable acid addition salts of the compounds described herein include, for example, salts of inorganic acids (such as hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, and sulfuric acid) and salts of organic acids (such as acetic acid, benzenesulfonic acid, benzoic acid, methanesulfonic acid, and p-toluenesulfonic acid). The compounds of this teaching, having an acidic group (such as a carboxylic acid), can form pharmaceutically acceptable salts with a pharmaceutically acceptable base. Suitable pharmaceutically acceptable basic salts include, for example, ammonium salts, alkali metal salts (such as sodium and potassium salts), and alkaline earth metal salts (such as magnesium and calcium salts). Compounds having a quaternary ammonium group also contain counter anions, such as chlorides, bromides, iodides, acetates, perchlorates, etc. Other examples of such salts include hydrochlorides, hydrobromic acids, sulfates, methanesulfonates, nitrates, benzoates, and salts with amino acids (such as glutamic acid).

[0031] The term “effective amount” or “therapeutic effective amount” refers to the amount of the compound described herein that is capable of evoking a desired or beneficial biological or medical response in a subject, for example, a dose between 0.01 and 100 mg / kg body weight / day.

[0032] 3. compound In the second embodiment, X is a halogen in the compound of formula I or its pharmaceutically acceptable salt, wherein the remaining variables are as described above for formula I.

[0033] In a third embodiment, in the compound of formula I or its pharmaceutically acceptable salt, X is selected from chlorine, bromine, fluorine, -SO2CH3, and -SOCH, wherein the remaining variables are as described above for formula I or the second embodiment. Alternatively, as part of a third embodiment, in the compound of formula I or its pharmaceutically acceptable salt, X is selected from chlorine, bromine, fluorine, and -SO2CH3, wherein the remaining variables are as described above for formula I or the second embodiment. In another alternative, as part of a third embodiment, in the compound of formula I or its pharmaceutically acceptable salt, X is chlorine, wherein the remaining variables are as described above for formula I or the second embodiment.

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

[0035] In the fifth embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. 2 Selected from hydrogen, halogen, (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 heterocyclic, -(C1-C4)alkyl[4- to 6-membered heterocyclic], -NR a R b-OR c -(C1-C4)alkyl OR c -C(O)R f -C(O)NR d R e -(C1-C4)alkyl NR d R e -(C1-C4)alkylC(O)R f -(C1-C4)alkylC(O)NR d R e and -SR g In each occurrence of (C3-C6) cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclic groups, each group is optionally represented by 1 to 3 groups selected from R 5 Substitution of groups; R a R b and R c Each group is 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 heterocyclic, -(C1-C4)alkyl[4- to 6-membered heterocyclic], wherein for each occurrence of (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclic, each group is optionally composed of 1 to 3 groups selected from R 6 Substitution of groups; R d and R e Each is independently selected from hydrogen, (C1-C4)alkyl, halo(C1-C4)alkyl, -(C1-C4)alkylOR', -(C1-C4)alkylNR'R'', -(C1-C4)alkylC(O)NR'R'', -(C1-C4)alkylC(O)R', -(C1-C4)alkylC(O)OR', -(C1-C4)alkylS(O)R', -(C1-C4)alkylS(O)2R', (C1-C4)alkylphenyl, phenyl, (C3-C6)cycloalkyl, -(C1-C4)alkyl[5 to 7-membered heteroaryl], 5 to 7-membered heteroaryl, 4 to 6-membered heterocyclic, -(C1-C4)alkyl[4 to 6-membered heterocyclic], -C(O)R', -C(O)OR', -C(O)NR'R'', -S(O)R' and -S(O)2R', wherein for each occurrence of (C3-C6)cycloalkyl, phenyl, 5 to 7-membered heteroaryl and 4 to 6-membered heterocyclic, the group is optionally composed of 1 to 3 groups selected from R 7 Substitution of the group; or R d and R eTogether with the nitrogen atom to which it is attached, they form each optionally composed of 1 to 3 atoms selected from R 7 substituted with 4- to 6-membered heterocyclic groups or 5- to 7-membered heteroaryl groups; R f The group 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 heterocyclic groups, wherein the phenyl, (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclic groups are each optionally composed of 1 to 3 groups selected from R 8 Substitution of groups; R a R 6 R 7 and R 8 Each is 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)2R'; and R' and R'' are each independently selected from hydrogen, (C1-C4)alkyl and (C3-C6)cycloalkyl, wherein the remaining variables are as described above for any of the second to fourth embodiments.

[0036] In the sixth embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. 2 Selected from hydrogen, halogen, (C1-C4)alkyl, (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, phenyl, 4- to 6-membered heterocyclic, -NR a R b -OR c -C(O)NR d R e -(C1-C4)alkyl NR d R e and -(C1-C4)alkylC(O)NR d R e In each occurrence of a 5- to 7-membered heteroaryl, phenyl, (C3-C6)cycloalkyl, and 4- to 6-membered heterocyclic group, each group is optionally represented by 1 to 3 groups selected from R 5 The group substitution, wherein the remaining variables are as described above for Formula I or any of the second to fifth embodiments. Alternatively, as part of a sixth embodiment, in the compound of Formula I or its pharmaceutically acceptable salt, R 2Selected from hydrogen, halogen, (C1-C4)alkyl, cyclopropyl, pyrazolyl, phenyl, pyrrolidinyl, piperazine, piperidinyl, -NR a R b -OR c -C(O)NR d R e -(C1-C4)alkyl NR d R e and -(C1-C4)alkylC(O)NR d R e The cyclopropyl, pyrazolyl, phenyl, pyrrolidinyl, piperazineyl, and piperidinyl groups are each optionally selected from R... 5 The group substitutions, wherein the remaining variables are as described above for Formula I or any of the second to fifth embodiments.

[0037] In the seventh embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. a R b and R c Each group is independently selected from hydrogen, (C1-C4)alkyl, -(C1-C4)alkyl [5- to 7-membered heteroaryl], phenyl, and -(C1-C4)alkylphenyl, 4- to 6-membered heterocyclic, -(C1-C4)alkyl [4- to 6-membered heterocyclic], wherein for each occurrence of 5- to 7-membered heteroaryl, 4- to 6-membered heterocyclic, and phenyl, each group is optionally composed of 1 to 3 groups selected from R 6 The group substitutions, wherein the remaining variables are as described above for Formula I or any of the second to sixth embodiments.

[0038] In the eighth embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. 5 and R 6 Each is independently selected from (C1-C4)alkyl and halo(C1-C4)alkyl, wherein the remaining variables are as described above for any of the second to seventh embodiments.

[0039] In the ninth embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. 2 Selected from hydrogen, chlorine, methyl, cyclopropyl, -OCH2CF3, , , , , , , , , , , , , , , , , , , , , and The remaining variables are as described above for Formula I or any of the second through eighth embodiments. Alternatively, as part of a ninth embodiment, in the compound of Formula I or its pharmaceutically acceptable salt, R 2 Selected from hydrogen, chlorine, and -OCH2CF3, wherein the remaining variables are as described above for Formula I or any of the second to eighth embodiments. In another alternative, as part of a ninth embodiment, R is selected from the compound of Formula I or a pharmaceutically acceptable salt thereof. 2 It is hydrogen, wherein the other variables are as described above for Formula I or any of the second to eighth embodiments.

[0040] In the tenth embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. 3 Selected from halogens, (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 heterocyclic, -(C1-C4)alkyl[4- to 9-membered heterocyclic], -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)alkyl NR g1 R h1 -(C1-C4)alkylC(O)R f1 -(C1-C4)alkylC(O)NR d1 R e1 and -SR g1 In each occurrence of (C3-C6) cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclic groups, each group is optionally represented by 1 to 3 groups selected from R 5a Substitution of groups; R a1 R b1 and R c1Each group is 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 heterocyclic, -(C1-C4)alkyl[4- to 6-membered heterocyclic], wherein for each occurrence of (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclic, each group is optionally composed of 1 to 3 groups selected from R 6a Substitution of groups; R d1 R e1 R g1 and R h1 Each is independently selected from 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 heterocyclic, -(C1-C4)alkyl[4 to 6-membered heterocyclic], -C(O)R' 1 -C(O)OR' 1 -C(O)NR' 1 R'' 1 -S(O)R' 1 and -S(O)2R' 1 In each occurrence of (C3-C6) cycloalkyl, phenyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclic groups, each group is optionally represented by 1 to 3 groups selected from R 7a Substitution of the group; or R d1 and R e1 Together with the nitrogen atom to which it is attached, they form each optionally composed of 1 to 3 atoms selected from R 7a substituted with 4- to 6-membered heterocyclic groups or 5- to 7-membered heteroaryl groups; R f1The group 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 heterocyclic groups, wherein the phenyl, (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclic groups are each optionally composed of 1 to 3 groups selected from R 8a Substitution of groups; R 5a R 6a R 7a and R 8a Each is independently selected from halogen, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, (C3-C6)cycloalkyl, cyano, -NR' 1 R'' 1 -(C1-C4)alkylNR' 1 R'' 1 -(C1-C4)alkylC(O)NR' 1 R'' 1 ,Oxyl,-(C1-C4)alkyl OR' 1 -C(O)R' 1 -S(O)R' 1 and -S(O)2R' 1 ;as well as R' 1 and R'' 1 Each is independently selected from hydrogen, (C1-C4)alkyl, (C3-C6)cycloalkyl and 4 to 7-membered heterocyclic groups, wherein the remaining variables are as described above for any of the second to ninth embodiments.

[0041] In the eleventh embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. 3 Selected from -(C1-C4)alkyl [4- to 9-membered heterocyclic], -(C1-C4)alkyl [5- to 7-membered heteroaryl], 4- to 6-membered heterocyclic, -(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)alkyl NR g1 R h1 The (C3-C6) cycloalkyl, 5- to 7-membered heteroaryl, 4- to 6-membered heterocyclic and 4- to 9-membered heterocyclic groups are each optionally composed of 1 to 3 groups selected from R 5aThe group substitution, wherein the remaining variables are as described above for Formula I or any of the second to tenth embodiments. Alternatively, in a compound of Formula I or a pharmaceutically acceptable salt thereof, R 3 Selected from -(C1-C4)alkyl OR c1 , 4 to 6-membered heterocyclic groups, -(C1-C4)alkyl [4 to 9-membered heterocyclic groups], -NR a1 R b1 -NHC(O)R c1 -C(O)R f1 -C(O)NR d1 R e1 -(C1-C4)alkyl NR g1 R h1 The 4- to 6-membered heterocyclic groups and the 4- to 9-membered heterocyclic groups are each optionally composed of 1 to 3 groups selected from R 5a The group substitution, wherein the remaining variables are as described above for Formula I or any of the second to tenth embodiments. In another embodiment, in the compound of Formula I or its pharmaceutically acceptable salt, R 3 Selected from -(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)alkyl NR g1 R h1 -(C1-C4)alkyl[piperazinyl], -(C1-C4)alkyl[piperidinyl], -(C1-C4)alkyl[morpholinyl], -(C1-C4)alkyl[pyrrolidinyl], -(C1-C4)alkyl[diazazyl], -(C1-C4)alkyl[azacyclobutyl], piperazinyl, and tetrahydropyridinyl, wherein the piperidinyl, morpholinyl, pyrrolidinyl, diazazyl, tetrahydropyridinyl, azacyclobutyl, and each occurrence of piperazinyl are optionally selected from 1 to 3 of R 5a The group substitutions, wherein the remaining variables are as described above for Formula I or any of the second to tenth embodiments.

[0042] In the twelfth embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. 5a Selected from (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' 1 The remaining variables are as described above for Formula I or any of the second to eleventh embodiments. Alternatively, as part of the twelfth embodiment, in the compound of Formula I or its pharmaceutically acceptable salt, R 5a The components are 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, wherein the remaining variables are as described above for any of the embodiments of Formula I or the second to eleventh embodiments.

[0043] In the thirteenth embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. c1 Selected from 5- to 7-membered heteroaryl and 4- to 6-membered heterocyclic groups, wherein each of the 5- to 7-membered heteroaryl and 4- to 6-membered heterocyclic groups is optionally composed of 1 to 3 groups selected from R 6a The group substitution, wherein the remaining variables are as described above for Formula I or any of the second to twelfth embodiments. Alternatively, as part of a thirteenth embodiment, in the compound of Formula I or its pharmaceutically acceptable salt, R c1 It is piperidinyl or pyridinyl, each optionally composed of 1 to 3 groups selected from R 6a The group substitutions, wherein the remaining variables are as described above for Formula I or any of the second to twelfth embodiments.

[0044] In the fourteenth embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. 6a It is a (C1-C4) alkyl group, wherein the remaining variables are as described above for any of the second to thirteenth embodiments.

[0045] In the fifteenth embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. a1 and R b1 Each group is independently selected from hydrogen, -(C1-C4)alkyl [5- to 7-membered heteroaryl] and 4- to 6-membered heterocyclic groups, wherein for each occurrence of the 5- to 7-membered heteroaryl and the 4- to 6-membered heterocyclic group, each group is optionally composed of 1 to 3 groups selected from R 6a The group substitution, wherein the remaining variables are as described above for Formula I or any of the second to fourteenth embodiments. Alternatively, as part of the fifteenth embodiment, in the compound of Formula I or its pharmaceutically acceptable salt, R a1 and R b1Each is independently selected from hydrogen and -(C1-C4)alkyl [5 to 7-membered heteroaryl], wherein the 5 to 7-membered heteroaryl group is optionally composed of 1 to 3 atoms selected from R 6a The group substitution, wherein the remaining variables are as described above for Formula I or any of the second to fourteenth embodiments. In another alternative, as part of the fifteenth embodiment, in the compound of Formula I or its pharmaceutically acceptable salt, R a1 It is hydrogen and R b1 It is -(C1-C4)alkyl[pyridinyl], wherein the pyridinyl group is optionally composed of 1 to 3 atoms selected from R 6a The group substitutions, wherein the remaining variables are as described above for Formula I or any of the second to fourteenth embodiments.

[0046] In the sixteenth embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. f1 It is optional to be selected from 1 to 3 R 8a The group substituted with a 4- to 6-membered heterocyclic group, wherein the remaining variables are as described above for any of the second to fifteenth embodiments of Formula I. Alternatively, as part of the sixteenth embodiment, in a compound of Formula I or a pharmaceutically acceptable salt thereof, R f1 It is optional to be selected from 1 to 3 R 8a The piperazine group is substituted with a group, wherein the remaining variables are as described above for any of the second to fifteenth embodiments.

[0047] In the seventeenth embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. 8a It is a (C1-C4) alkyl group, wherein the remaining variables are as described above for any of the second to sixteenth embodiments.

[0048] In the eighteenth embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. d1 and R e1 Each is independently selected from hydrogen, -(C1-C4)alkyl NR' 1 R'' 1 (C1-C4)alkyl [5- to 7-membered heteroaryl] and 4- to 6-membered heterocyclic groups, wherein the 5- to 7-membered heteroaryl and the 4- to 6-membered heterocyclic groups are each optionally composed of 1 to 3 groups selected from R 7a The group substitution, wherein the remaining variables are as described above for Formula I or any of the second to seventeenth embodiments. Alternatively, as part of the eighteenth embodiment, in the compound of Formula I or its pharmaceutically acceptable salt, R d1 and R e1Each is independently selected from hydrogen, -(C1-C4)alkylN[(C1-C4)alkyl]2, (C1-C4)alkyl[pyridinyl] and piperidinyl, wherein the pyridinyl and piperidinyl are each optionally composed of 1 to 3 elements selected from R 7a The group substitutions, wherein the remaining variables are as described above for Formula I or any of the second to seventeenth embodiments.

[0049] In the nineteenth embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. g1 and R h1 Each is independently selected from 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 heterocyclic, wherein the phenyl, 5- to 7-membered heteroaryl and 4- to 6-membered heterocyclic are each optionally composed of 1 to 3 groups selected from R 7a The group substitution, wherein the remaining variables are as described above for Formula I or any of the second to eighteenth embodiments. Alternatively, as part of the nineteenth embodiment, in the compound of Formula I or its pharmaceutically acceptable salt, R g1 and R h1 Each of the following is independently selected from hydrogen, (C1-C4)alkyl, halo(C1-C4)alkyl, -(C1-C4)alkyl[pyridyl], -(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 the phenyl, pyridinyl, piperidinyl, pyrrolidinyl and morpholinyl are each optionally selected from 1 to 3 of R 7a The group substitution, wherein the remaining variables are as described above for Formula I or any of the second to eighteenth embodiments. In another alternative, as part of the nineteenth embodiment, in the Formula I compound or its pharmaceutically acceptable salt, R 7a The group is selected from (C1-C4)alkyl, halo(C1-C4)alkyl, halogen, (C1-C4)alkoxy and cyano, wherein the remaining variables are as described above for any of the embodiments of Formula I or the second to the eighteenth embodiments.

[0050] In the twentieth embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. 3 Selected from –CH2NHCH3, –CH2N(CH3)2, , -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, , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and The remaining variables are as described above for Equation I or any of the second to nineteenth embodiments.

[0051] In the twenty-first embodiment, A is N in the compound of formula I or its pharmaceutically acceptable salt, wherein the remaining variables are as described above for formula I. Alternatively, as part of the twenty-first embodiment, Y is N in the compound of formula I or its pharmaceutically acceptable salt, wherein the remaining variables are as described above for formula I. Alternatively, as part of the twenty-first embodiment, Z is N in the compound of formula I or its pharmaceutically acceptable salt, wherein the remaining variables are as described above for formula I.

[0052] In the twenty-second embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. 2 It is hydrogen, wherein the other variables are as described above for Formula I or the twenty-first embodiment.

[0053] In the twenty-third embodiment, X is a halogen in the compound of formula I or a pharmaceutically acceptable salt thereof, wherein the remaining variables are as described above for formula I or the twenty-first or twenty-second embodiment. Alternatively, as part of the twenty-third embodiment, X is bromine or chlorine in the compound of formula I or a pharmaceutically acceptable salt thereof, wherein the remaining variables are as described above for formula I or the twenty-first or twenty-second embodiment.

[0054] In the twenty-fourth embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. 1 It is a haloalkyl (C1-C4)alkyl or cyanoalkyl, wherein the remaining variables are as described above for any of the twenty-first to twenty-third embodiments. Alternatively, as part of the twenty-fourth embodiment, in a compound of formula I or a pharmaceutically acceptable salt thereof, R 1 It is –CH2CH3 or –CH2CN, wherein the remaining variables are as described above for formula I or any of the twenty-first to twenty-third embodiments.

[0055] In the twenty-fifth embodiment, R is in a compound of formula I or a pharmaceutically acceptable salt thereof. 3 It is an optionally substituted alkyl group, wherein the remaining variables are as described above for Formula I or any of the twenty-first to twenty-fourth embodiments. Alternatively, as part of the twenty-fifth embodiment, in a compound of Formula I or a pharmaceutically acceptable salt thereof, R 3 It is a -(C1-C4)alkyl heterocyclic group or a -(C1-C4)alkyl NR g1 R h1 , where R g1 and R h1Each is independently selected from hydrogen and (C1-C4) alkyl groups, wherein the remaining variables are as described above for Formula I or any of the twenty-first to twenty-fourth embodiments. Alternatively, as part of the twenty-fifth embodiment, in a compound of Formula I or a pharmaceutically acceptable salt thereof, R 3 It is a -(C1-C4)bicyclic heterocyclic group or a -(C1-C4)alkyl NR g1 R h1 , where R g1 and R h1 Each is independently selected from hydrogen and (C1-C4) alkyl groups, wherein the remaining variables are as described above for Formula I or any of the twenty-first to twenty-fourth embodiments. Alternatively, as part of the twenty-fifth embodiment, in a compound of Formula I or a pharmaceutically acceptable salt thereof, R 3 Selected from , , , , , and The remaining variables are as described above for Equation I or any of the twenty-first to twenty-fourth embodiments.

[0056] Compounds having Formula I are further disclosed in the examples and are included in this disclosure. This includes pharmaceutically acceptable salts and neutral forms.

[0057] 4. Uses, formulations and application The compounds and compositions described herein are generally used to modulate the activity of TP53. In some aspects, the compounds, pharmaceutically acceptable salts, and pharmaceutical compositions described herein are covalent modifiers of Y220C. In some aspects, the compounds, pharmaceutically acceptable salts, and pharmaceutical compositions described herein stabilize TP53. In some aspects, the compounds, pharmaceutically acceptable salts, and pharmaceutical compositions described herein restore the function of wild-type tumor suppressor protein p53 (WT TP53).

[0058] In some aspects, the compounds and pharmaceutical compositions described herein can be used to treat conditions related to TP53 function. In some aspects, the compounds and pharmaceutical compositions described herein can be used to treat conditions that respond to TP53 functional activation. In some aspects, the compounds and pharmaceutical compositions described herein can be used to treat conditions that respond to TP53 functional recovery, for example, conditions resulting from protein loss of function due to mutations such as the Y220C mutation.

[0059] In some aspects, the compounds and pharmaceutical compositions described herein can be used to treat cancer. In some aspects, the compounds and pharmaceutical compositions described herein can be used to treat cancers expressing TP53 mutants (e.g., cancers carrying the Y220C mutation).

[0060] The compounds, salts, and compositions of the present invention treat specific cancers, including but not limited to solid tumors, hematologic malignancies, ovarian cancer, esophageal cancer, colorectal 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.

[0061] Also provided is the use of the compounds described herein or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the disclosed compounds or pharmaceutically acceptable salts thereof, in the preparation of a medicament for treating the condition. Further provided are the compounds described herein or pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the disclosed compounds or pharmaceutically acceptable salts thereof, for treating the condition.

[0062] In some aspects, the pharmaceutical compositions described herein are formulated for administration to patients in need of such compositions. The pharmaceutical compositions described herein can be administered orally, parenterally, via inhalation spray, topically, rectally, nasally, buccally, vaginally, or via an implanted reservoir. As used herein, the term "parenterical" 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. The sterile injectable formulations of the compositions described herein can be aqueous or oily suspensions. These suspensions can be formulated using suitable dispersants or wetting agents and suspending agents according to techniques known in the art.

[0063] In some respects, the pharmaceutical composition is administered orally.

[0064] The specific dosage and treatment regimen for any given patient will depend on a number of factors, including the activity of the specific compound used, age, weight, general health condition, sex, diet, timing of administration, excretion rate, drug combination, physician's judgment, and the severity of the specific disease being treated. The amount of the compounds described herein in the composition also depends on the specific compounds in the drug combination.

[0065] Example Chemical synthesis The following representative embodiments are intended to help illustrate the present disclosure and are not intended to, nor should they be construed as, limiting the scope of the invention.

[0066] Option 1: 4-Chloropyridine-pyrrolopyrimidine, such as S10, can be prepared by a nine-step method starting with a suitable fluorocyanopyridine, S1. S1 is alkylated with ethyl bromoacetate to provide intermediate S2, which can be cyclized using Boc2O and DMAP to provide indole intermediate S3. S3 is condensed with DMF-DMA, followed by treatment with ammonia, to provide pyridine-pyrrolopyrimidine intermediate S5. Chlorination with POCl3 followed by alkylation with ethyl trifluoromethanesulfonate yields intermediate S7. Treatment of S7 with sodium methoxide yields intermediate S8, which can be converted to the target compound, such as S10, via a two-step sequential process using Suzuki coupling with potassium ((dimethylamino)methyl)trifluoroborate and treatment with POCl3.

[0067] Option 2: 4-Chloropyridine-pyrrolopyrimidine, such as S15, can also be prepared using a four-step method starting with intermediate S11. S11 is alkylated with ethyl trifluoromethanesulfonate to give intermediate S12. Suzuki coupling is performed using 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborane, followed by ozonolysis to provide the aldehyde intermediate S14. Reductive amination is then performed using an amine to provide the target compound, such as S15.

[0068] Option 3: 4-Chloropyridopyrrolopyrimidines such as S23 can be prepared by a seven-step method starting from a suitable fluorocyanopyridine S16. S16 is alkylated with ethyl bromoacetate to provide intermediate S17, which can be cyclized using Boc2O and DMAP to provide indole intermediate S18. S18 is condensed with DMF-DMA, followed by treatment with ammonia, to provide pyridopyrrolopyrimidine intermediate S20. Chlorination with POCl3 followed by alkylation with ethyl trifluoromethanesulfonate yields intermediate S22. S22 is Suzuki coupled using potassium ((dimethylamino)methyl)trifluoroborate to provide the target compound such as S23.

[0069] abbreviation: ACN = Acetonitrile AcOH = Acetic acid Boc2O = ditert-butyl dicarbonate DCM = dichloromethane DCE = 1,2-dichloroethane DIEA = N,N-diisopropylethylamine DMAP = 4-Dimethylaminopyridine DPPA = Diphenylphosphoazide DMF = dimethylformamide DMF-DMA = N,N-dimethylformamide dimethyl acetal DPPA = Diphenylphosphoazide EtOAc = Ethyl acetate EtOH = ethanol IPA = Isopropanol MeOH = Methanol TEA = Triethylamine TFA = Trifluoroacetic acid THF = Tetrahydrofuran TMSCl = Trimethylchlorosilane TMSBr = Trimethylbromosilane Example 1: 1-(4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d] Pyrimidin-8-yl)-N,N-dimethylformamide Scheme 1, Step 1: Ethyl 2-[(6-bromo-4-cyano-3-pyridyl)amino] Add 2-aminoethyl ethyl hydrochloride (2.78 g, 19.90 mmol, 2 equivalents) and DIEA (3.86 g, 29.85 mmol, 5.20 mL, 3 equivalents) to a solution of 2-bromo-5-fluoro-pyridin-4-carboxynitrile (2.0 g, 9.95 mmol, 1 equivalent) in ACN (25 mL). Stir the mixture at 60 °C for 12 hours. Cool the reaction to room temperature and dilute with EtOAc (30.0 mL). Wash the mixture with H₂O and brine, dry with Na₂SO₄, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography (petroleum ether containing 10-25% ethyl acetate) to give the title compound (1.95 g, 69% yield) as a yellow solid.

[0070] LCMS [M+1, M+3] = 284.0, 286.0.

[0071] Scheme 1, Step 2: 3-Amino-5-bromo-1H-pyrrolo[2,3-c]pyridine-1,2-1-(tert-butyl)2-ethyldicarboxylic acid Ester: Add Boc₂O (1.34 g, 6.12 mmol, 1.20 equivalence), TEA (710 μL, 5.10 mmol, 1 equivalence), and DMAP (62 mg, 510 μmol, 0.1 equivalence) to a solution of ethyl 2-[(6-bromo-4-cyano-3-pyridyl)amino]ethyl acetate (1.45 g, 5.10 mmol, 1 equivalence) in DCM (15 mL). Stir the mixture at room temperature for 2 hours. Dilute the reaction mixture with DCM (30.0 mL). Wash the mixture with H₂O and brine, dry it with Na₂SO₄, filter, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography (petroleum ether containing 10-25% ethyl acetate) to give the title compound (2.58 g, crude) as a yellow solid.

[0072] LCMS [M+1, M+3] = 384.1, 386.0.

[0073] Scheme 1, Step 3: 5-Bromo-3-[(E)-dimethylaminomethyleneamino]-pyrrolo[2,3-c]pyridine-1,2- Ethyl dicarboxylate: To a solution of 3-amino-5-bromo-1H-pyrrolo[2,3-c]pyridine-1,2-1-(tert-butyl)-2-ethyl dicarboxylic acid ester (2.3 g, 5.99 mmol, 1 equivalent) in DMF (25 mL), 1,1-dimethoxy-N,N-dimethyl-methylamine (1.03 mL, 7.78 mmol, 1.3 equivalent) was added. The mixture was then stirred at 100 °C for 4 hours. The reaction was cooled to room temperature and diluted with EtOAc (30.0 mL). The mixture was washed with H2O and brine, dried over Na2SO4, filtered, and concentrated to give the title compound (2.68 g, crude) as a green solid.

[0074] LCMS [M+1, M+3] = 439.0, 441.0.

[0075] Scheme 1, Step 4: 8-bromo-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidin-4-ol: NH3·H2O (20 mL) was added to a solution of ethyl 5-bromo-3-[(E)-dimethylaminomethyleneamino]-pyrrolo[2,3-c]pyridine-1,2-dicarboxylic acid (2.2 g, 5.01 mmol, 1 equivalent) in MeOH (20 mL). The mixture was then stirred at 70 °C for 16 hours. The reaction was cooled to room temperature and the mixture was concentrated under reduced pressure to give the title compound (1.45 g, crude) as a brown solid.

[0076] LCMS [M+1, M+3] = 265.1, 267.1.

[0077] Scheme 1, Step 5: 8-bromo-4-chloro-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine: A solution of 8-bromo-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidin-4-ol (1.45 g, 5.47 mmol, 1 equivalent) in POCl3 (15 mL) was stirred at 110 °C for 16 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was then ground with ACN (5 mL) at room temperature to give the title compound (1.12 g, yield 61%, purity 84%) as a brown solid.

[0078] LCMS [M+1, M+3] = 282.9, 284.9.

[0079] Scheme 1, Step 6: 8-bromo-4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]-pyrrolo [3,2-d]pyrimidine: To a solution of 8-bromo-4-chloro-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine (1.12 g, 3.95 mmol, 1 equivalent) in DMF (12 mL), 2,2,2-trifluoroethyltrifluoromethanesulfonate (1.83 g, 7.90 mmol, 2 equivalents) and K₂CO₃ (1.09 g, 7.90 mmol, 2 equivalents) were added. The reaction mixture was stirred at room temperature for 20 hours. The reaction mixture was diluted with EtOAc (30.0 mL), washed with H₂O and brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether containing 10-25% ethyl acetate) to give the title compound (850 mg, 56% yield) as a white solid.

[0080] LCMS [M+1, M+3] = 364.8, 366.9.

[0081] Scheme 1, Step 7: 8-bromo-4-methoxy-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrole [3,2-d]pyrimidine: Sodium methoxide (39.4 mg, 219 μmol, 0.8 equivalent; 30% methanol solution) was added to a MeOH solution of 8-bromo-4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]-pyrrolo[3,2-d]pyrimidine (100 mg, 273.57 μmol, 1 equivalent). The mixture was then stirred at 40 °C for 1 hour. The reaction mixture was cooled to room temperature and diluted with ethyl acetate (30.0 mL). The mixture was washed with H2O and brine, dried over Na2SO4, filtered, and concentrated to give the title compound as a white solid (95 mg, 96% yield).

[0082] LCMS [M+1, M+3] = 360.9, 362.9.

[0083] Scheme 1, Step 8: 1-(4-methoxy-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]-pyrrole [3,2-d]pyrimidin-8-yl)-N,N-dimethylformamide: To a solution of 8-bromo-4-methoxy-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine (95 mg, 263 μmol, 1 equivalent) in 2-methylbut-2-ol (1 mL) and H₂O (0.25 mL), potassium ((dimethylamino)methyl)trifluoroborate (86.8 mg, 526 μmol, 2 equivalents), Cs₂CO₃ (171 mg, 526 μmol, 2 equivalents), and [1,1'-bis(di-tert-butylphosphine)ferrocene]palladium(II) dichloride (17.1 mg, 26.3 μmol, 0.1 equivalents) were added. The suspension was degassed with nitrogen and then stirred at 80 °C for 8 hours. The reaction was cooled to room temperature and diluted with EtOAc (30.0 mL). The organic layer was washed with H2O, then with brine, dried with Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO2, 10:1 DCM: MeOH) to give the title compound (65 mg, 73% yield) as a brown solid.

[0084] LCMS [M+1] = 340.1.

[0085] Scheme 1, Step 9: 1-(4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3, 2-d]pyrimidin-8-yl)-N,N-dimethylformamide: A mixture of 1-(4-methoxy-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]-pyrrolo[3,2-d]pyrimidin-8-yl)-N,N-dimethylformamide (15 mg, 44.21 μmol, 1 equivalent) and POCl3 (0.5 mL) was degassed with nitrogen and then stirred at 110 °C for 20 h. The reaction mixture was cooled to room temperature and then a saturated aqueous solution of NaHCO3 was added dropwise. The mixture was extracted with EtOAc (3 x 10.0 mL). The combined organic layers were washed with brine (2 x 10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO2, 10:1 DCM: MeOH) to give the title compound (8.2 mg, 53.48% yield) as a brown solid.

[0086] LCMS [M+1, M+3] = 343.9, 345.9.

[0087] 1H NMR (400 MHz, chloroform-) d ) δ 9.12 (s, 1H), 9.04 (s, 1H), 8.40 (s, 1H), 5.45 (q, J = 8.0 Hz, 2H), 3.97 (br s, 2H), 2.49 (br s, 6H).

[0088] Example 2: 1-(4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d] Pyrimidin-8-yl)-N-methylmethylamine Scheme 2, Step 1: 8-bromo-4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]-pyrrolo [3,2-d]pyrimidine: To a solution of 8-bromo-4-chloro-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine (1.12 g, 3.95 mmol, 1 equivalent) in DMF (12 mL), 2,2,2-trifluoroethyltrifluoromethanesulfonate (1.83 g, 7.90 mmol, 2 equivalents) and K₂CO₃ (1.09 g, 7.90 mmol, 2 equivalents) were added. The reaction mixture was stirred at room temperature for 20 hours. The reaction mixture was diluted with EtOAc (30.0 mL), washed with H₂O and brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether containing 10-25% ethyl acetate) to give the title compound (850 mg, 56% yield) as a white solid.

[0089] LCMS [M+1, M+3] = 364.8, 366.9.

[0090] Scheme 2, Step 2: 4-chloro-5-(2,2,2-trifluoroethyl)-8-vinyl-5H-pyrido[4',3':4,5]pyrrole [3,2-d]pyrimidine: The mixture of 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborane (195 μL, 1.15 mmol, 1.2 equivalents), 8-bromo-4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine (350 mg, 958 μmol, 1 equivalent), Cs2CO3 (343 mg, 1.05 mmol, 1.1 equivalents) and [1,1'-bis(di-tert-butylphosphine)ferrocene]palladium(II) dichloride (62.4 mg, 95.8 μmol, 0.1 equivalents) in H2O (4 mL) and 2-methylbut-2-ol (16 mL) was degassed with nitrogen and then stirred at 80 °C for 2 hours under a nitrogen atmosphere. The reaction mixture was cooled to room temperature, poured into H2O (5 mL), and extracted with ethyl acetate (3 x 5 mL). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO2, petroleum ether:ethyl acetate 3:1) to give the title compound as a white solid (150 mg, 50% yield).

[0091] LCMS: [M+1, M+3] = 313.1, 315.2.

[0092] Scheme 2, Step 3: 4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d] Pyrimidine-8-carbaldehyde: To a solution of 4-chloro-5-(2,2,2-trifluoroethyl)-8-vinyl-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine (150 mg, 480 μmol, 1 equivalent) in dioxane (6 mL) and H₂O (3 mL), K₂O₄·H₂O (4.4 mg, 11.9 μmol, 0.025 equivalent), NaIO₄ (410 mg, 1.92 mmol, 4 equivalent), and 2,6-dimethylpyridine (112 μL, 959 μmol, 2 equivalent) were added. The mixture was stirred at room temperature for 2 hours. The reaction mixture was poured into H₂O (5 mL) and extracted with ethyl acetate (3 x 5 mL). The combined organic extracts were washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether containing 1-50% ethyl acetate) to give the title compound as a white solid (140 mg, 93% yield).

[0093] LCMS: [M+1, M+3] = 315.1, 317.1.

[0094] Scheme 2, Step 4: 1-(4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]-pyrrolo[3, 2-d]pyrimidin-8-yl)-N-methylmethylamine To a solution of 4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidin-8-carboxaldehyde (90 mg, 286 μmol, 1 equivalent) and methylamine (286.0 μL, 592 μmol, 2 equivalent; 2M, THF) in DCE (0.1 mL), NaBH(OAc)3 (152 mg, 715 μmol, 2.5 equivalent) and AcOH (33 μL, 572 μmol, 2 equivalent) were added, and the mixture was stirred at room temperature for 30 min. At this point, additional NaBH(OAc)3 (151.5 mg, 715.1 μmol, 2.5 equivalent) was added, and the mixture was stirred at room temperature for 2 h. The reaction mixture was poured into H2O (5 mL) and extracted with ethyl acetate (3 x 5 mL). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residues were purified by preparative HPLC (column: Phenomenex Luna C18 75 mm x 30 mm x 3 µm; mobile phase: water containing 5–35% acetonitrile (+0.04% HCl)) to give the title compound (36.7 mg, 35% yield; HCl salt) as a white solid.

[0095] LCMS: [M+1, M+3] = 330.1, 332.1.

[0096] 1 H NMR (400 MHz, DMSO- d 6 ) δ 9.60 (s, 1H), 9.45 (m, 1H), 9.11 (s, 1H), 8.52 (s, 1H), 5.93 (q, J = 8.8 Hz, 2H), 4.49 (t, J = 6.0 Hz, 2H), 2.63 (t, J = 5.6Hz, 3H).

[0097] Example 3: 2-(4-chloro-8-((8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)methyl)-5H-pyridine [4',3':4,5]pyrrolo[3,2-d]pyrimidin-5-yl)acetonitrile Scheme 2, Step 1: 2-(8-bromo-4-chloro-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidin-5-yl) Acetonitrile: A solution of 8-bromo-4-chloro-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine (100.0 mg, 352.7 μmol, 1 equivalent) in DMF (3.0 mL) was cooled to 0 °C, and Cs₂CO₃ (459.6 mg, 1.4 mmol, 4.0 equivalent) and 2-chloroacetonitrile (45 μL, 705 μmol, 2.0 equivalent) were added. The solution was warmed to room temperature and then stirred for 12 hours. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (3 x 20 mL). The combined organic extracts were washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether containing 0-25% ethyl acetate) to give the title compound (100.0 mg, 88% yield) as a yellow solid.

[0098] LCMS [M +1, M +3] = 322.0, 324.0.

[0099] Scheme 2, Step 2: 2-(4-chloro-8-vinyl-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine-5- Acetonitrile (based on) To a solution of 2-(8-bromo-4-chloro-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidin-5-yl)acetonitrile (50.0 mg, 155.0 μmol, 1.0 equivalent) in H₂O (1.0 mL) and 2-methylbut-2-ol (4.0 mL), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborane (19.1 mg, 124.0 μmol, 0.8 equivalent), Cs₂CO₃ (56 mg, 171 μmol, 1.1 equivalent), and [1,1'-bis(di-tert-butylphosphine)ferrocene]palladium(II) dichloride (10.1 mg, 15.5 μmol, 0.1 equivalent) were added. The reaction mixture was stirred at 80 °C for 1 hour under a nitrogen atmosphere. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (3 x 20 mL). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether containing 0-100% ethyl acetate) to give the title compound as a yellow solid (40.0 mg, 96% yield).

[0100] LCMS [M +1, M+3] = 270.1, 272.1.

[0101] Scheme 2, Step 3: 2-(4-chloro-8-formyl-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine-5- Acetonitrile (based on) To a solution of 2-(4-chloro-8-vinyl-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidin-5-yl)acetonitrile (100.0 mg, 370.8 μmol, 1.0 equivalent) in 1,4-dioxane (10.0 mL) and H₂O (5.0 mL), 2,6-dimethylpyridine (86 μL, 742 μmol, 2 equivalents), NaIO₄ (317 mg, 1.4 mmol, 4.0 equivalents), and K₂O₅SO₄·H₂O (3.4 mg, 9.2 μmol, 0.025 equivalents) were added. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (3 x 20 mL). The combined organic extracts were washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether containing 0-100% ethyl acetate) to give the title compound as a yellow solid (60.0 mg, 60% yield).

[0102] LCMS [M +1, M+3] = 272.1, 274.1.

[0103] Scheme 2, Step 4: 2-(4-chloro-8-((8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)methyl)-5H- Pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidin-5-yl)acetonitrile: Add 2-(4-chloro-8-formyl-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidin-5-yl)acetonitrile (10.0 mg, 36.8 μmol, 1.0 equivalent) and isopropoxytitanium(IV) (65.1 μL, 220.8 μmol, 6.0 equivalent) to a THF (0.5 mL) solution of 8-methyl-3,8-diazabicyclo[3.2.1]octane (10.9 mg, 55.2 μmol, 1.5 equivalent) and isopropoxytitanium(IV) (65.1 μL, 220.8 μmol, 6.0 equivalent). Stir the mixture at room temperature for 30 min, then add NaBH(OAc)3 (23.4 mg, 110.4 μmol, 3 equivalent). Stir the reaction mixture at room temperature for 12 h. Dilute the reaction mixture with water (10 mL) and extract with dichloromethane (3 x 10 mL). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residues were purified by preparative HPLC (column: Phenomenex luna C18 100 mm x 40 mm x 5 µm; mobile phase: water containing 1-25% acetonitrile (+0.04% HCl)) to give the title compound (15.3 mg, 54% yield; HCl salt) as a white solid.

[0104] LCMS [M +1, M+3] = 382.2, 384.1.

[0105] 1 H NMR (400 MHz, methanol-) d 4 ) δ 9.84 (s, 1H), 9.17 (s, 1H), 8.92 (s, 1H),6.17 (s, 2H), 4.30 (s, 2H), 4.00 (s, 2H), 3.19 - 3.07 (m, 4H), 2.84 (s, 3H),2.39 - 2.29 (m, 4H).

[0106] Example 4: 4-chloro-8-(((8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)methyl)-5-(2,2,2- Trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine Scheme 2, Step 3: 4-chloro-8-(((8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)methyl)-5-(2, 2,2-Trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine: At room temperature, 8-methyl-3,8-diazabicyclo[3.2.1]octane (232.6 mg, 1.4 mmol, 1.5 equivalence; HCl salt) and AcOH (109 μL, 1.9 mmol, 2.0 equivalence) were added to a DCE (3.0 mL) solution of 4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine-8-carboxaldehyde (300.0 mg, 953.4 μmol, 1.0 equivalence) and DCE (3.0 mL). The mixture was stirred at room temperature for 30 min, then NaBH(OAc)3 (606 mg, 2.8 mmol, 3.0 equivalence) was added, and the solution was stirred at room temperature for 12 h. The reaction mixture was diluted with water (10.0 mL) and extracted with dichloromethane (3 x 10.0 mL). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residues were purified by silica gel column chromatography (ethyl acetate containing 0-15% methanol) to give the title compound as a yellow solid (180.0 mg, 44% yield).

[0107] LCMS[M +1, M+3] = 425.0, 427.0.

[0108] 1 H NMR (400 MHz, DMSO- d 6 ) δ 9.41 (s, 1H), 9.06 (s, 1H), 8.25 (s, 1H), 5.85 (q, J= 8.8 Hz, 2H), 3.80 (s, 2H), 3.13 - 3.04 (m, 2H), 2.59 (d, J = 10.0Hz, 2H), 2.38 (d, J = 10.0 Hz, 2H), 2.23 - 2.15 (m, 3H), 1.96 - 1.87 (m, 2H), 1.87 - 1.78 (m, 2H).

[0109] Example 5: 4-Chloro-8-((2-methyl-2,7-diazaspiro[3.5]non-7-yl)methyl)-5-(2,2,2-trifluoro Ethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine Scheme 2, Step 3: 4-chloro-8-((2-methyl-2,7-diazaspiro[3.5]non-7-yl)methyl)-5-(2,2,2- Trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine: TEA (44 μL, 318 μmol, 2 equivalents) was added to a DCE (0.5 mL) solution of 4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidin-8-carboxaldehyde (50 mg, 159 μmol, 1 equivalent) and 2-methyl-2,7-diazaspiro[3.5]nonane dihydrochloride (33.9 mg, 158.9 μmol, 1 equivalent), and the mixture was stirred at room temperature for 30 min. Then NaBH(OAc)3 (84.2 mg, 397.3 μmol, 2.5 equivalents) was added, and the mixture was stirred at room temperature for 2 h. The reaction mixture was poured into H2O (5 mL) and extracted with ethyl acetate (3 x 5 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: Waters Xbridge BEH C18 100 mm x 30 mm x 10 µm; mobile phase: water containing 15-60% acetonitrile (+10 mMNH4HCO3)) to give the title compound as a white solid (9.7 mg, yield 6.68%).

[0110] LCMS: [M+1, M+3] = 439.2, 441.1.

[0111] 1 H NMR (400 MHz, methanol-) d 4 ) δ = 9.27 (s, 1H), 8.99 (s, 1H), 8.39 (s, 1H), 5.76 (q, J= 8.4 Hz, 2H), 3.85 (s, 2H), 3.14 (s, 4H), 2.50 (br s, 4H), 2.38 (s,3H), 1.81 (br t, J = 5.2 Hz, 4H).

[0112] Example 6: 4-chloro-8-((7-methyl-2,7-diazaspiro[4.4]non-2-yl)methyl)-5-(2,2,2-trifluoro Ethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine Scheme 2, Step 3: 4-chloro-8-((7-methyl-2,7-diazaspiro[4.4]non-2-yl)methyl)-5-(2,2,2- Trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine: AcOH (200 µL 3.4 mmol, 7.0 equivalence) was added to a DCE (2.0 mL) solution of 4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidin-8-carboxaldehyde (155 mg, 493 μmol, 1.0 equivalence) and 2-methyl-2,7-diazaspiro[4.4]nonane (104 mg, 739 μmol, 1.5 equivalence), and the mixture was stirred at room temperature for 30 min. Then, NaBH(OAc)3 (313.2 mg, 1.4 mmol, 3.0 equivalence) was added to the reaction mixture, and the mixture was stirred at room temperature for 1 h. The reaction mixture was poured into H2O (5 mL) and extracted with ethyl acetate (3 x 5 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 (Phenomenex Luna C18 75 mm x 30 mm x 3 µm; mobile phase: water containing 1-43% acetonitrile (+0.04% HCl)). A second purification was performed by preparative HPLC (Agela DuraShell NH2 150 mm x 30 mm x 5 µm; mobile phase: heptane containing 50-95% ethanol) to give the title compound as a brown solid (9.6 mg, yield 4.3%).

[0113] LCMS [M+1, M+3] = 439.2, 441.1.

[0114] 1 H NMR (400 MHz, MeOD) δ 9.27 (s, 1H), 9.00 (s, 1H), 8.41 (s, 1H), 5.77 (q, J= 8.4 Hz, 2H), 4.03 (s, 2H), 2.84 - 2.71 (m, 5H), 2.69 - 2.62 (m,3H), 2.43 (s, 3H), 2.05 - 1.87 (m, 4H).

[0115] Example 7: 4-Chloro-8-((8-methyl-2,8-diazaspiro[4.5]dec-2-yl)methyl)-5-(2,2,2-trifluoro Ethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine Scheme 2, Step 3: 4-chloro-8-((8-methyl-2,8-diazaspiro[4.5]dec-2-yl)methyl)-5-(2,2,2- Trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine: To a DCE (1.0 mL) solution of 4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidin-8-carboxaldehyde (100.0 mg, 317.8 μmol, 1.0 equivalent), 8-methyl-2,8-diazaspiro[4.5]decane (73.5 mg, 476.7 μmol, 1.5 equivalent) and AcOH (0.1 mL) were added, and the mixture was stirred at room temperature for 30 min. Then, NaBH(OAc)3 (202.0 mg, 953.4 μmol, 3.0 equivalent) was added, and the reaction mixture was stirred at room temperature for 1 h. The reaction mixture was poured into H2O (5 mL) and extracted with ethyl acetate (3 x 5 mL). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by preparative HPLC (Agela DuraShell NH2 150mm x 30mm x 5µm; mobile phase: [heptane-EtOH]; gradient: 50% - 95% B, duration 10.0 min) to give the title compound as a brown solid (16.8 mg, yield 10.9%).

[0116] LCMS [M+1, M+3] = 453.2, 455.1.

[0117] 1 H NMR (400 MHz, MeOD) δ 9.27 (s, 1H), 9.00 (s, 1H), 8.40 (s, 1H), 5.76 (q, J = 8.4 Hz, 2H), 3.99 (s, 2H), 2.76 (t, J = 6.8 Hz, 2H), 2.57 (s, 2H), 2.44 (s, 3H), 2.27 (s, 3H), 1.73 - 1.67 (m, 7H).

[0118] Example 8: 8-((3,8-diazabicyclo[3.2.1]oct-3-yl)methyl)-4-chloro-5-(2,2,2-trifluoroethyl) )-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine Scheme 2, Step 3: 3-((4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3, 2-d]pyrimidin-8-yl)methyl)-3,8-diazabicyclo[3.2.1]oct-8-carboxylic acid tert-butyl ester: To a DCE (0.5 mL) solution of 4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidin-8-carboxaldehyde (50 mg, 159 μmol, 1 equivalent) and 3,8-diazabicyclo[3.2.1]octyl-8-carboxylic acid tert-butyl ester (67 mg, 318 μmol, 2 equivalents), NaBH(OAc)3 (84.20 mg, 397.27 μmol, 2.5 equivalents) and AcOH (19.09 mg, 317.81 μmol, 18.19 μL, 2 equivalents) was added, and the mixture was stirred at room temperature for 30 minutes. Additional NaBH(OAc)3 (84.2 mg, 397 μmol, 2.5 equivalents) was added, and the mixture was stirred for another 2 hours. The reaction mixture was poured into H2O (5 mL) and extracted with ethyl acetate (3 x 5 mL). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative TLC (SiO2, petroleum ether:ethyl acetate 1:1) to give the title compound as a white solid (80 mg, 99% yield).

[0119] LCMS: [M+1, M+3] = 511.3, 513.3.

[0120] Scheme 2, Step 3a: 8-((3,8-diazabicyclo[3.2.1]oct-3-yl)methyl)-4-chloro-5-(2,2,2-tri) (Fluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine: To a flask containing 3-((4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidin-8-yl)methyl)-3,8-diazabicyclo[3.2.1]octyl-8-carboxylic acid tert-butyl ester (80 mg, 156.57 μmol, 1 equivalent), HCl (8.00 mL, 204 equivalents; 4 M, dioxane) was added. The mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 75 mm x 30 mm x 3 µm; mobile phase: water containing 5–35% acetonitrile (+0.04% HCl)) to give the title compound (38.1 mg, 54% yield; HCl salt) as a white solid.

[0121] LCMS: [M+1, M+3] = 411.1, 413.1.

[0122] 1 H NMR (400 MHz, DMSO- d 6 ) δ 9.54 (s, 1H), 9.51 - 9.26 (m, 1H), 9.11 (s, 1H), 8.58 (br s, 1H), 5.91 (q, J = 8.4 Hz, 2H), 4.28 (br s, 2H), 4.09 (brs, 2H), 3.19 (br s, 4H), 2.22 (br d, J = 6.8 Hz, 2H), 2.09 - 1.90 (m, 2H).

[0123] Example 9: 4-Fluoro-8-((8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)methyl)-5-(2,2,2- Trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine 4-Fluoro-8-((8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)methyl)-5-(2,2,2-trifluoroethyl )-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine: At room temperature, 1,4,7,10,13,16-hexaoxane (1.2 mg, 4.7 μmol, 0.1 equivalent) and tetramethylammonium chloride (515.9 μg, 4.7 μmol, 0.1 equivalent) were added to a solution of cesium fluoride (21.4 mg, 141.2 μmol, 5.2 μL, 3.0 equivalent). Then, 4-chloro-8-((8-methyl-3,8-diazabicyclo[3.2.1]oct-3-yl)methyl)-5-(2,2,2-trifluoroethyl)-5H-pyrido[4',3':4,5]pyrrolo[3,2-d]pyrimidine (20.0 mg, 47.0 μmol, 1.0 equivalent) was added in solution of acetonitrile (0.3 mL), and the reaction mixture was stirred at 45 °C for 25 min. The reaction mixture was cooled to room temperature, diluted with water (10 mL), and extracted with dichloromethane (3 x 10.0 mL). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residues were purified by preparative HPLC (column: Waters XbridgePrep OBD C18 150 mm x 40 mm x 10 μm; mobile phase: water containing 20–50% acetonitrile (+10 mM NH4HCO3)) to give the title compound as a white solid (5.0 mg, yield 5.1%).

[0124] LCMS [M+1] = 409.1.

[0125] 1H NMR (400 MHz, acetonitrile- d 3 ) δ 9.14 (s, 1H), 8.83 (s, 1H), 8.29 (s, 1H), 5.28 (q, J = 8.8 Hz, 2H), 3.81 (s, 2H), 3.04 (s, 2H), 2.62 (dd, J = 2.4, 10.4 Hz, 2H), 2.41 (d, J = 10.0 Hz, 2H), 2.20 (s, 3H), 1.92 - 1.80 (m, 4H).

[0126] Example 10: 1-(4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[2',3':4,5]pyrrolo[3,2-d] Pyrimidin-8-yl)-N,N-dimethylformamide Option 3, Step 1: Ethyl 2-[(6-bromo-2-cyano-3-pyridyl)amino]: DIEA (1.30 mL, 7.46 mmol, 3 equivalents) was added to a solution of 6-bromo-3-fluoro-pyridine-2-carboxynitrile (500 mg, 2.49 mmol, 1 equivalent) and ethyl aminoethyl hydrochloride (694 mg, 4.98 mmol, 2 equivalents) in acetonitrile (5 mL). The mixture was stirred at 65 °C for 12 hours. The reaction was carried out three times on this scale, and the combined solutions were then post-processed. The reaction mixture was poured into H2O (50 mL) and extracted with ethyl acetate (3 x 50 mL). The combined organic extracts were washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether containing 1-10% ethyl acetate) to give the title compound (2.7 g, crude) as a white solid.

[0127] LCMS: [M+1, M+3] = 284.5, 286.4.

[0128] Scheme 3, Step 2: 3-Amino-5-bromo-1H-pyrrolo[3,2-b]pyridine-1,2-1-(tert-butyl)2-ethyldicarboxylic acid Ester: DMAP (39 mg, 317 μmol, 0.1 equivalence), Boc₂O (830 mg, 3.80 mmol, 1.2 equivalence), and DIEA (441 μL, 3.17 mmol, 1 equivalence) were added to a solution of ethyl 2-[(6-bromo-2-cyano-3-pyridyl)amino]ethyl acetate (900 mg, 3.17 mmol, 1 equivalence) in DCM (10 mL). The mixture was stirred at room temperature for 2 hours. The reaction was carried out three times on this scale, and the combined solutions were then post-processed. The reaction mixture was poured into H₂O (50 mL) and extracted with ethyl acetate (3 x 50 mL). The combined organic extracts were washed with brine (50 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether containing 1-10% ethyl acetate) to give the title compound (2.6 g, 71% yield) as a white solid.

[0129] LCMS: [M+1, M+3] = 384.2, 386.3.

[0130] Scheme 3, Step 3: (E)-5-bromo-3-(((dimethylamino)methylene)amino)-1H-pyrrolo[3,2-b]pyrrolo Pyridine-1,2-1-(tert-butyl)2-ethyl dicarboxylic acid ester: DMF-DMA (944 μL, 7.11 mmol, 1.3 equivalents) was added to a DMF (20 mL) solution of 3-amino-5-bromo-1H-pyrrolo[3,2-b]pyridine-1,2-1-(tert-butyl)-2-ethyl dicarboxylic acid ester (2.1 g, 5.47 mmol, 1 equivalent). The mixture was stirred at 100 °C for 4 hours. The reaction mixture was cooled to room temperature, poured into H2O (20 mL), and extracted with ethyl acetate (3 x 20 mL). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to give the title compound (2.2 g, 92% yield) as a white solid.

[0131] Scheme 3, Step 4: 8-bromo-5H-pyrido[2',3':4,5]pyrrolo[3,2-d]pyrimidin-4-ol: To a solution of (E)-5-bromo-3-(((dimethylamino)methylene)amino)-1H-pyrrolo[3,2-b]pyridine-1,2-1-(tert-butyl)2-ethyl dicarboxylic acid ester (1.8 g, 4.10 mmol, 1 equivalent) in MeOH (18 mL), NH3·H2O (18.00 mL, 140.22 mmol, 34.2 equivalents; 30% aqueous solution) was added. The mixture was stirred at 70 °C for 72 hours. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give the title compound (1.1 g, crude) as a white solid.

[0132] Scheme 3, Step 5: 8-bromo-4-chloro-5H-pyrido[2',3':4,5]pyrrolo[3,2-d]pyrimidine: POCl3 (1.20 mL, 12.9 mmol, 34 equivalents) was added to a solution of 8-bromo-5H-pyrido[2',3':4,5]pyrrolo[3,2-d]pyrimidin-4-ol (100 mg, 377 μmol, 1 equivalent). The mixture was stirred at 110 °C for 12 hours. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give the title compound (100 mg, crude) as a white solid.

[0133] LCMS: [M+1, M+3] = 284.9, 286.9.

[0134] Scheme 3, Step 6: 8-bromo-4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[2',3':4,5]pyrrolo [3,2-d]pyrimidine: K₂CO₃ (293 mg, 2.12 mmol, 3 equivalents) was added to a DMF (3 mL) solution of 2,2,2-trifluoroethyltrifluoromethanesulfonate (327 mg, 1.41 mmol, 2 equivalents) and 8-bromo-4-chloro-5H-pyrido[2',3':4,5]pyrrolo[3,2-d]pyrimidine (200 mg, 705 μmol, 1 equivalent). The mixture was stirred at room temperature for 2 hours. The reaction mixture was poured into H₂O (5 mL) and extracted with ethyl acetate (3 x 5 mL). The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether containing 1-10% ethyl acetate) to give the title compound (300 mg, crude) as a white solid.

[0135] LCMS: [M+1, M+3] = 321.0, 322.9.

[0136] Scheme 3, Step 7: 1-(4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[2',3':4,5]pyrrolo[3, 2-d]pyrimidin-8-yl)-N,N-dimethylformamide: A mixture of 8-bromo-4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[2',3':4,5]pyrrolo[3,2-d]pyrimidine (270 mg, 739 μmol, 1 equivalent), potassium ((dimethylamino)methyl)trifluoroborate (244 mg, 1.48 mmol, 2 equivalents), Cs₂CO₃ (481.33 mg, 1.4 mmol, 2 equivalents), [1,1'-bis(di-tert-butylphosphine)ferrocene]palladium(II) dichloride (48.14 mg, 73.86 μmol, 0.1 equivalents) in 2-methylbut-2-ol (3 mL) and H₂O (0.8 mL) was degassed with nitrogen, and then the mixture was stirred at 80 °C for 2 hours. The reaction mixture was cooled to room temperature, diluted with H₂O, and extracted with EtOAc (3 x 20 mL). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residues were purified by preparative HPLC (column: Phenomenex luna C18 80 mm x 30 mm x 3 µm; mobile phase: water containing 1–30% acetonitrile (+0.04% HCl)) to give the title compound as a white solid (14.4 mg, yield 5.1%; HCl salt).

[0137] LCMS: [M+1, M+3] = 344.1, 346.1.

[0138] 1 H NMR (400 MHz, DMSO- d 6 ) δ 9.09 (s, 1H), 8.68 (d, J = 8.4 Hz, 1H), 8.08(d, J = 8.8 Hz, 1H), 5.85 (q, J = 8.8 Hz, 2H), 4.70 (br s, 2H), 2.87 (s, 6H).

[0139] Example 11: 1-(4-chloro-5-(2,2,2-trifluoroethyl)-5H-pyrido[3',2':4,5]pyrrolo[3,2-d] Pyrimidin-8-yl)-N,N-dimethylformamide Example 11 was prepared using the same sequence as Scheme 3 described in Example 10, but in step 1, 5-bromo-2-fluoro-pyridin-3-onitrile was used instead of 6-bromo-3-fluoro-pyridin-2-onitrile.

[0140] LCMS [M+1, M+3] = 344.1, 346.1.

[0141] 1 H NMR (400 MHz, chloroform-) d ) δ 8.98 (s, 1H), 8.76 (d, J = 1.8 Hz, 1H), 8.65(d, J = 1.8 Hz, 1H), 5.57 (d, J = 8.0 Hz, 2H), 3.71 (s, 2H), 2.34 (s, 6H).

[0142] Y220C TP53 TR-FRET binding probe, BP1, 5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a- Hexahydrothiophene[3,4-d]imidazol-4-yl]-N-[2-[2-[2-[2-[3-methoxy-4-[3-[4-(tetrahydropyran-4-ylamino)] [2,2,2-trifluoroethyl]indol-2-yl]prop-2-ynylamino]phenyl]sulfonylamino]ethoxy]ethoxy] Preparation of ethoxy[ethyl]pentanamide BP1 synthesis, step 1a, 2-iodo-4-nitro-1-(2,2,2-trifluoroethyl)indole: At 0 °C, NaH (2.0 g, 52.0 mmol, 5.0 equivalent; 60.0%, dispersed in oil) was added in portions to a THF (20 mL) solution of 2-iodo-4-nitro-1H-indole (3.0 g, 10.4 mmol, 1.0 equivalent) and stirred at 0 °C for 30 min. At 0 °C, 2,2,2-trifluoroethyltrifluoromethanesulfonate (9.6 g, 41.0 mmol, 4.0 equivalent) was added in portions to the reaction mixture. The mixture was stirred at 25 °C for 2 h. The reaction mixture was quenched with H₂O (20 mL), and the resulting mixture was partitioned between EtOAc (200 mL) and H₂O (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 Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether: EtOAc 15:1 to 12:1) to give the title compound (5.0 g, crude) as a yellow solid.

[0143] 1 ¹H 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).

[0144] BP1 synthesis, step 2a, 2-iodo-1-(2,2,2-trifluoroethyl)indole-4-amine:Fe (717 mg, 12.8 mmol, 2.5 equivalents) and NH4Cl (687 mg, 12.8 mmol, 2.5 equivalents) were added to a solution of 2-iodo-4-nitro-1-(2,2,2-trifluoroethyl)indole (1.9 g, 5.1 mmol, 1.0 equivalent) in EtOH (20.0 mL) and H2O (5.0 mL). The mixture was stirred at 80 °C for 2 h. The reaction solution was filtered through a diatomaceous earth mat, and the filtrate was partitioned between EtOAc (200 mL) and H2O (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 Na2SO4, filtered, and concentrated under reduced pressure to give the title compound (1.7 g, 97% yield) as a yellow solid.

[0145] LCMS [M+1] = 341.1.

[0146] BP1 synthesis, step 3a, 2-iodo-N-tetrahydropyran-4-yl-1-(2,2,2-trifluoroethyl)indole-4-amine: Chloro(trimethyl)silane (559.8 μL, 4.4 mmol, 2.5 equivalence) and tetrahydropyran-4-one (648.1 μL, 7.0 mmol, 4.0 equivalence) were added to a solution of 2-iodo-1-(2,2,2-trifluoroethyl)indole-4-amine (600 mg, 1.7 mmol, 1.0 equivalence) in 10 mL of DMF. The mixture was stirred at 0 °C for 2 h. Borane-tetrahydrofuran complex (1 M, 8.8 mL, 5.0 equivalence) was added to the mixture under N2, and the resulting mixture was stirred at 0–20 °C for 12 h. The reactants were poured into a saturated aqueous solution of NH4Cl (1.5 mL) and 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.

[0147] 1 H 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 = 9.4 Hz, 1H), 2.24 - 2.01 (m, 2H).

[0148] LCMS [M+1] = 425.1.

[0149] BP1 synthesis, step 1b, N-[2-[2-[2-[2-[(3-methoxy-4-nitro-phenyl)sulfonylamino]ethoxy [B-ethyl] tert-butyl carbamate: TEA (1.6 mL, 11.9 mmol, 5.0 equivalence) and 3-methoxy-4-nitrobenzenesulfonyl chloride (600 mg, 2.4 mmol, 1.0 equivalence) were added to a DCM (15 mL) solution of N-[2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]-ethyl]carbamate (906 mg, 3.1 mmol, 1.3 equivalence). The mixture was stirred at 15 °C for 1 hour. The reaction mixture was concentrated under reduced pressure, and the resulting residue was partitioned between EtOAc (50 mL) and H2O (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 Na2SO4, filtered, and concentrated under reduced pressure to give the title compound (1.1 g, 91% yield) as a brown oil.

[0150] 1 H NMR (400 MHz, DMSO- d 6 ) 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).

[0151] LCMS [M+1] = 408.2. BP1 synthesis, step 2b, N-[2-[2-[2-[2-[(4-amino-3-methoxy-phenyl)sulfonylamino]ethoxy] [B-ethyl] tert-butyl carbamate:Fe (1.1 g, 19.7 mmol, 5.0 equivalent) and NH4Cl (1.1 g, 19.7 mmol, 5.0 equivalent) were added to a solution of N-[2-[2-[2-[2-[(3-methoxy-4-nitro-phenyl)sulfonylamino]ethoxy]ethoxy]ethoxy]-ethyl]carbamate tert-butyl ester (2.0 g, 3.9 mmol, 1.0 equivalent) in EtOH (12.0 mL) and H2O (3.0 mL). The mixture was stirred at 80 °C for 1 hour. The suspension was filtered through a diatomaceous earth mat, and the mat 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 H2O (30 mL). The aqueous layer was then extracted again with EtOAc (2 × 50 mL). The combined organic layers were washed with brine (50 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the title compound (1.5 g, 80% yield) in a brown oil.

[0152] 1 H NMR (400 MHz, DMSO- d 6 ) 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).

[0153] LCMS [M+1] = 378.3.

[0154] BP1 synthesis, step 3b, N-[2-[2-[2-[2-[3-methoxy-4-(3-trimethylsilylprop-2-ynylamino)] [Phenyl]-sulfonylamino]ethoxy]ethoxy]ethoxy]ethyl]tert-butyl carbamate:Add 3-trimethylsilylpropion-2-acetylenealdehyde (396 mg, 3.1 mmol, 1.0 equivalent) to a solution of N-[2-[2-[2-[2-[(4-amino-3-methoxy-phenyl)sulfonylamino]ethoxy]ethoxy]ethoxy]-ethyl]carbamate tert-butyl ester (1.5 g, 3.1 mmol, 1.0 equivalent) in DCM (15 mL) and acetic acid (3 mL). Stir the mixture at 35 °C for 17 hours, then add sodium triacetoxyborohydride (2.6 g, 12.5 mmol, 4.0 equivalent) to the mixture. Stir the mixture at 35 °C for 17 hours. Dilute the reaction mixture with EtOAc (50 mL) and H2O (30 mL), and extract the aqueous layer with EtOAc (2 × 50 mL). The combined organic extracts were washed with brine (50 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex luna C18 (250 mm × 70 mm × 15 μm); mobile phase: water containing 40–75% ACN (+NH₄HCO₃ modifier)) to give the title compound (480 mg, 26% yield) as a white solid.

[0155] 1 H NMR (400 MHz, DMSO- d 6 ) 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).

[0156] LCMS [M+1] = 488.4.

[0157] BP1 synthesis, step 4b, N-[2-[2-[2-[2-[[3-methoxy-4-(prop-2-ynylamino)phenyl]sulfonyl] [Amino]-ethoxy]ethoxy]ethoxy]ethyl]tert-butyl carbamate: K₂CO₃ (272.7 mg, 1.9 mmol, 2.0 equivalent) was added to a solution of N-[2-[2-[2-[[3-methoxy-4-(3-trimethylsilylprop-2-ynylamino)phenyl]sulfonylamino]ethoxy]ethoxy]ethoxy]ethyl]-tert-butyl carbamate (580.0 mg, 986.7 μmol, 1.0 equivalent) in MeOH (6.0 mL). The mixture was stirred at 20 °C for 1 hour. The reaction mixture was partitioned between EtOAc (50 mL) and H₂O (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 Na₂SO₄, filtered, and concentrated under reduced pressure to give the title compound (400 mg, 79% yield) as an off-white oil.

[0158] 1 H NMR (400 MHz, DMSO- d 6 ) 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).

[0159] LCMS [M+1] = 416.3.

[0160] BP1 synthesis, step 5b, 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 [B-ethyl] tert-butyl carbamate:Under a nitrogen atmosphere, triphenylphosphine dichloropalladium (27.2 mg, 38.7 μmol, 0.1 equivalent) and cuprous(I) iodide (7.4 mg, 38.8 μmol, 0.1 equivalent), 2-iodo-N-tetrahydropyran-4-yl-1-(2,2,2-trifluoroethyl)indole-4-amine (165 mg, 388 μmol, 1.0 equivalent) and triethylamine (162 µL, 1.1 mmol, 3.0 equivalent) were added to a solution of N-[2-[2-[2-[2-[2-[[3-methoxy-4-(prop-2-ynylamino)phenyl]sulfonylamino]ethoxy]-ethoxy]ethoxy]ethyl]carbamate tert-butyl (200 mg, 388 μmol, 1.0 equivalent) in ACN (2.0 mL). The mixture was stirred at 70 °C for 2 hours. The reaction mixture was partitioned between EtOAc (10 mL) and H2O (10 mL), and the aqueous layer was extracted with EtOAc (2 × 10 mL). The combined organic extracts were washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the title compound as a white solid (200 mg, 64% yield).

[0161] 1 H NMR (400 MHz, DMSO- d 6 ) 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).

[0162] LCMS [M+1] = 812.3.

[0163] 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]benzenesulfonyl amine: TFA (0.8 mL) was added to a solution of 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]tert-butyl carbamate (200 mg, 246 μmol, 1.0 equivalent) in DCM (0.6 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 for the next step.

[0164] LCMS [M+1] = 712.4.

[0165] BP1 synthesis, step 7b, TR-FRET binding probe, BP1, 5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6, 6a-Hexahydrothiophene[3,4-d]imidazol-4-yl]-N-[2-[2-[2-[2-[3-methoxy-4-[3-[4-(tetrahydropyran-4-) [2,2,2-trifluoroethyl]indol-2-yl]prop-2-ynylamino]phenyl]-sulfonylamino]ethoxy]ethyl [Oxygen]ethoxy]ethyl]pentanamide:Add TEA (28.4 mg, 281 μmol, 39.1 μL, 5.0 equivalent) and 5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothiophene[3,4-d]imidazol-4-yl]valerate (2,5-dioxopyrrolidine-1-yl) ester (19.1 mg, 56.2 μmol, 1.0 equivalent) to a DCM (1 mL) solution of N-[2-[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 equivalent)] in 5 mL of DCM. The mixture was stirred at 20°C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a residue, which was purified by preparative HPLC (column: Phenomenex Luna C18 200 mm × 40 mm × 10 μm; mobile phase: water containing 35–70% ACN (+ formic acid modifier)) to give the title compound as a white solid (11.2 mg, yield 21%).

[0166] 1 H NMR (400 MHz, DMSO- d 6 ) 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).

[0167] LCMS [M+1] = 938.3.

[0168] Bioassay The efficacy of the compound (IC50) was evaluated in a TR-FRET binding assay measuring the binding of the biotinylated small molecule probe BP1 to the Y220C TP53 mutant DBD. 50 Specifically, 5 μL of a mixture (consisting of 32 nM BP1 and 48 nM streptavidin-d2 (Cisbio) at pH 7.2, 10 mM KH2PO4, 150 mM NaCl, 0.01% BSA, 0.01% Tween-20, and 0.1 mM TCEP) was added to a 384-well plate containing the test compound in 60 nL DMSO (0.6% fc .DMSO (v / v)) for repeated 10-point dose-response titrations. An additional 5 μL of mixture was added to a 384-well plate. This mixture consisted of 10 nM Y220C TP53 DBD (expressed in *E. coli*, His-TEV-P89-T312-FLAG; Uniprot ID P04637-1) in 10 mM KH2PO4 at pH 7.2, 0.3 nM Mab anti-FLAG M2-Tb cavitation compound (Perkin Elmer), 150 mM NaCl, 0.01% BSA, 0.01% Tween-20, and 0.1 mM TCEP. The mixture was incubated at 20 °C. The TR-FRET reaction was monitored after 4 h and 24 h of incubation with the BP1 mixture and the compound mixture. The plates were read using an EnVision plate reader (Perkin Elmer) at Ex / Em 615 / 665. To determine the potency (IC50) of the test compound...50 (Competitively binding to TP53 Y220C protein in the presence of the biotinylated probe BP1), the TR-FRET ratio was normalized to the mean ratio (0% inhibition) of the DMSO control wells and the mean minimum ratio (100% inhibition) obtained using the 5 μmol BP1 positive control compound. The dose-response of the test compounds was fitted using a nonlinear regression with 4-parameter fitting, providing IC50. 50 Values ​​and nHill slope. The results are shown in Table 1 below and expressed in the ranges 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 >60.0 μM; % inhibition <10%.

[0169] The TR-FRET assay described above can be used to evaluate the binding affinity of reversible and covalent TP53 Y220C ligands, stabilizers, and correctors, and can also distinguish between them. To validate the TR-FRET assay described above, we characterized TP53 Y220C ligands, stabilizers, and correctors previously described in the scientific literature. (By Joerger et al.) ACS Chem. Biology Compounds PK9301 and PK9323, described in (2020, 15, 657-668), have been shown by X-ray crystallography to bind to TP53 Y220C, and their stability in the mutant protein was determined by differential scanning fluorometry (DSF). In the aforementioned TR-FRET binding assay, their IC50 values ​​were [missing information]. 50 =1.5 μM and 4.1 μM respectively. Jeorger, Boeckler et al. ( J. Am. Chem. Soc. Compound PK5196, described in 2012, 134, 6810-6818, has been shown by X-ray crystallography to bind to TP53 Y220C, and its stability to the mutant protein has been determined by differential scanning fluorometry (DSF), nuclear magnetic resonance (NMR), and isothermal calorimetry (ITC). In the aforementioned TR-FRET assays, IC5196 showed a high efficiency of 100%. 50 =9.7 μM. PK9301, PK9323, and PK5196 are all reversible TP53 Y220C ligands / stabilizers, and therefore exhibit time-independent activity in the above TR-FRET assays. The same or nearly identical IC50 values ​​were observed at 4 h and 24 h time points. 50Value. Covalent ligands / modifiers typically exhibit time-dependent activity because covalent modification of a protein produces a persistent, cumulative effect as the reaction with the target protein proceeds. Compounds of Formula I typically exhibit a time-dependent shift of the TR-FRET binding probe BP1 from TP53 Y220C, thus distinguishing it from reversible TP53 Y220C ligands / stabilizers / correctors. We believe that, in certain cases, covalent modification of mutant TP53Y220C may lead to more persistent stabilization of the mutant protein and result in a more robust recovery of wild-type TP53 function relative to reversible ligands. This covalent mechanism of action may be advantageous for small molecule therapies targeting medical conditions such as cancer associated with or attributable to the TP53 Y220C mutant protein.

[0170] Table 1

[0171] While we have described many embodiments, it will be apparent that our basic embodiments can be modified to provide other embodiments utilizing the compounds and methods of the present invention. Therefore, it should be understood that the scope of the invention is defined by the appended claims, and not by the specific embodiments shown by way of example.

[0172] All references cited in this application (including bibliographic references, granted 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 have the meanings commonly understood by one of ordinary skill in the art.

Claims

1. A compound having formula I: (I)； Or its pharmaceutically acceptable salt, in which R 1 Selected from alkyl groups with optional substitution; R 2 Selected from hydrogen, halogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted heterocyclic, -NR a R b -OR c -NHC(O)R c -C(O)NR d R e -C(O)R f and -SR g ; R 3 Selected from hydrogen, halogen, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted heterocyclic, -NR a1 R b1 -OR c1 -NHC(O)R c1 -C(O)NR d1 R e1 -C(O)R f1 and -SR g1 ; X is selected from halogens, -S(O)2 alkyl groups, and -S(O) alkyl groups; The variables that are N if one of A, Y, or Z is not N are -CH-; R a R a1 R b R b1 R c and R c1 Each is independently selected from hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted aryl and optionally substituted heterocyclic groups; R d and R e Each is independently selected from hydrogen, optionally substituted alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted heterocyclic, -C(O)R', -C(O)OR', -C(O)NR'R'', -S(O)R' and -S(O)2R'; or R d and R e Together with the nitrogen atom to which it is attached, it forms an optionally substituted heterocyclic group or an optionally substituted heteroaryl group; R d1 and R e1 Each is independently selected from hydrogen, optionally substituted alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, optionally substituted heterocyclic, -C(O)R' 1 -C(O)OR' 1 -C(O)NR' 1 R'' 1 -S(O)R' 1 and -S(O)2R' 1 Or R d1 and R e1 Together with the nitrogen atom to which it is attached, it forms an optionally substituted heterocyclic group or an optionally substituted heteroaryl group; R f and R f1 Each is independently selected from hydrogen, optionally substituted alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted heteroaryl, and optionally substituted heterocyclic; and R'、R' 1 , R'' and R'' 1 Each is independently selected from hydrogen and optionally substituted (C1-C4) alkyl groups.

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

3. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein Y is N.

4. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein Z is N.

5. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein R 2 It is hydrogen.

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

7. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein X is bromine or chlorine.

8. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein R 1 It is a haloalkyl (C1-C4) or cyanoalkyl (C1-C4) alkyl.

9. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 8, wherein R 1 It is -CH2CH3 or -CH2CN.

10. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, wherein R 3 It is an alkyl group that is optionally substituted.

11. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 10, wherein R 3 It is a -(C1-C4)alkyl heterocyclic group or a -(C1-C4)alkyl NR g1 R h1 , where R g1 and R h1 Each is independently selected from hydrogen and (C1-C4) alkyl groups.

12. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 11, wherein R 3 It is a -(C1-C4)bicyclic heterocyclic group or a -(C1-C4)alkyl NR g1 R h1 , where R g1 and R h1 Each is independently selected from hydrogen and (C1-C4) alkyl groups.

13. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 12, wherein R 3 Selected from , , , , , and .

14. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein X is selected from chlorine, bromine, fluorine, -SO2(C1-C3)alkyl and -SO(C1-C3)alkyl.

15. The compound of claim 1 or claim 14, or a pharmaceutically acceptable salt thereof, wherein X is selected from chlorine, bromine, fluorine, -SO2CH3, and -SOCH3.

16. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1, 14 and 15, wherein X is selected from chlorine, bromine, fluorine and -SO2CH3.

17. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 and 14 to 16, wherein X is chlorine.

18. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 17, wherein R 1 It is selected from (C1-C4)alkyl, halo(C1-C4)alkyl, cyano(C1-C4)alkyl and -(C1-C4)alkyl(C3-C6)cycloalkyl.

19. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 18, wherein R 1 Selected from (C1-C4)alkyl and halo(C1-C4)alkyl.

20. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 19, wherein R 1 Selected from -CH2CH3, -CH2CF3, -CH2CF2CH3 and -CH2CF2CF3.

21. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 20, wherein R 1 It is -CH2CF3.

22. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 21, wherein... R 2 Selected from hydrogen, halogen, (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 heterocyclic, -(C1-C4)alkyl[4- to 6-membered heterocyclic], -NR a R b -OR c -(C1-C4)alkyl OR c -C(O)R f -C(O)NR d R e -(C1-C4)alkyl NR d R e -(C1-C4)alkylC(O)R f -(C1-C4)alkylC(O)NR d R e and -SR g In each occurrence of (C3-C6) cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclic groups, each group is optionally represented by 1 to 3 groups selected from R 5 Substitution of groups; R a R b and R c Each group is 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 heterocyclic, -(C1-C4)alkyl[4- to 6-membered heterocyclic], wherein for each occurrence of (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclic, each group is optionally composed of 1 to 3 groups selected from R 6 Substitution of groups; R d and R e Each is independently selected from hydrogen, (C1-C4)alkyl, halo(C1-C4)alkyl, -(C1-C4)alkylOR', -(C1-C4)alkylNR'R'', -(C1-C4)alkylC(O)NR'R'', -(C1-C4)alkylC(O)R', -(C1-C4)alkylC(O)OR', -(C1-C4)alkylS(O)R', -(C1-C4)alkylS(O)2R', (C1-C4)alkylphenyl, phenyl, (C3-C6)cycloalkyl, -(C1-C4)alkyl[5 to 7-membered heteroaryl], 5 to 7-membered heteroaryl, 4 to 6-membered heterocyclic, -(C1-C4)alkyl[4 to 6-membered heterocyclic], -C(O)R', -C(O)OR', -C(O)NR'R'', -S(O)R' and -S(O)2R', wherein for each occurrence of (C3-C6)cycloalkyl, phenyl, 5 to 7-membered heteroaryl and 4 to 6-membered heterocyclic, the group is optionally composed of 1 to 3 groups selected from R 7 Substitution of the group; or R d and R e Together with the nitrogen atom to which it is attached, they form each optionally composed of 1 to 3 atoms selected from R 7 substituted with 4- to 6-membered heterocyclic groups or 5- to 7-membered heteroaryl groups; R f The group 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 heterocyclic groups, wherein the phenyl, (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclic groups are each optionally composed of 1 to 3 groups selected from R 8 Substitution of groups; R a R 6 R 7 and R 8 Each is 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)2R'; and R' and R'' are each independently selected from hydrogen, (C1-C4)alkyl, and (C3-C6)cycloalkyl.

23. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 22, wherein R 2 Selected from hydrogen, halogen, (C1-C4)alkyl, (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, phenyl, 4- to 6-membered heterocyclic, -NR a R b -OR c -C(O)NR d R e -(C1-C4)alkyl NR d R e and -(C1-C4)alkylC(O)NR d R e In each occurrence of a 5- to 7-membered heteroaryl, phenyl, (C3-C6)cycloalkyl, and 4- to 6-membered heterocyclic group, each group is optionally represented by 1 to 3 groups selected from R 5 The group is substituted.

24. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 23, wherein R 2 Selected from hydrogen, halogen, (C1-C4)alkyl, cyclopropyl, pyrazolyl, phenyl, pyrrolidinyl, piperazine, piperidinyl, -NR a R b -OR c -C(O)NR d Re, -(C1-C4)alkyl NR d R e and -(C1-C4)alkylC(O)NR d R e The cyclopropyl, pyrazolyl, phenyl, pyrrolidinyl, piperazineyl, and piperidinyl groups are each optionally selected from R... 5 The group is substituted.

25. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 24, wherein R a R b and R c Each group is independently selected from hydrogen, (C1-C4)alkyl, -(C1-C4)alkyl [5- to 7-membered heteroaryl], phenyl, and -(C1-C4)alkylphenyl, 4- to 6-membered heterocyclic, -(C1-C4)alkyl [4- to 6-membered heterocyclic], wherein for each occurrence of 5- to 7-membered heteroaryl, 4- to 6-membered heterocyclic, and phenyl, each group is optionally composed of 1 to 3 groups selected from R 6 The group is substituted.

26. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 25, wherein R 5 and R 6 Each is independently selected from (C1-C4)alkyl and halo(C1-C4)alkyl.

27. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 26, wherein R 2 Selected from hydrogen, chlorine, methyl, cyclopropyl, -OCH2CF3, , , , , , , , , , , , , , , , , , , , , and .

28. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 27, wherein R 2 Selected from hydrogen, chlorine and -OCH2CF3.

29. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 28, wherein R 2 It is hydrogen.

30. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 29, wherein R 3 Selected from halogens, (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 heterocyclic, -(C1-C4)alkyl[4- to 9-membered heterocyclic], -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)alkyl NR g1 R h1 -(C1-C4)alkylC(O)R f1 -(C1-C4)alkylC(O)NR d1 R e1 and -SR g1 In each occurrence of (C3-C6) cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclic groups, each group is optionally represented by 1 to 3 groups selected from R 5a Substitution of groups; R a1 R b1 and R c1 Each group is 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 heterocyclic, -(C1-C4)alkyl[4- to 6-membered heterocyclic], wherein for each occurrence of (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, phenyl, and 4- to 6-membered heterocyclic, each group is optionally composed of 1 to 3 groups selected from R 6a Substitution of groups; R d1 R e1 R g1 and R h1 Each is independently selected from 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 heterocyclic, -(C1-C4)alkyl[4 to 6-membered heterocyclic], -C(O)R' 1 -C(O)OR' 1 -C(O)NR' 1 R'' 1 -S(O)R' 1 and -S(O)2R' 1 In each occurrence of (C3-C6) cycloalkyl, phenyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclic groups, each group is optionally represented by 1 to 3 groups selected from R 7a Substitution of the group; or R d1 and R e1 Together with the nitrogen atom to which it is attached, they form each optionally composed of 1 to 3 atoms selected from R 7a substituted with 4- to 6-membered heterocyclic groups or 5- to 7-membered heteroaryl groups; R f1 The group 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 heterocyclic groups, wherein the phenyl, (C3-C6)cycloalkyl, 5- to 7-membered heteroaryl, and 4- to 6-membered heterocyclic groups are each optionally composed of 1 to 3 groups selected from R 8a Substitution of groups; R 5a R 6a R 7a and R 8a Each is independently selected from halogen, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, (C3-C6)cycloalkyl, cyano, -NR' 1 R'' 1 -(C1-C4)alkylNR' 1 R'' 1 -(C1-C4)alkylC(O)NR' 1 R'' 1 ,Oxyl,-(C1-C4)alkyl OR' 1 -C(O)R' 1 -S(O)R' 1 and -S(O)2R' 1 ;as well as R' 1 and R'' 1 Each is independently selected from hydrogen, (C1-C4)alkyl, (C3-C6)cycloalkyl and 4 to 7-membered heterocyclic groups.

31. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 30, wherein R 3 Selected from -(C1-C4)alkyl [4- to 9-membered heterocyclic], -(C1-C4)alkyl [5- to 7-membered heteroaryl], 4- to 6-membered heterocyclic, -(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)alkyl NR g1 R h1 The (C3-C6) cycloalkyl, 5- to 7-membered heteroaryl, 4- to 6-membered heterocyclic and 4- to 9-membered heterocyclic groups are each optionally composed of 1 to 3 groups selected from R 5a The group is substituted.

32. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 31, wherein R 3 Selected from -(C1-C4)alkyl OR c1 , 4 to 6-membered heterocyclic groups, -(C1-C4)alkyl [4 to 9-membered heterocyclic groups], -NR a1 R b1 -NHC(O)R c1 -C(O)R f1 -C(O)NR d1 R e1 -(C1-C4)alkyl NR g1 R h1 The 4- to 6-membered heterocyclic groups and the 4- to 9-membered heterocyclic groups are each optionally composed of 1 to 3 groups selected from R 5a The group is substituted.

33. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 32, wherein R 3 Selected from -(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)alkyl NR g1 R h1 -(C1-C4)alkyl[piperazinyl], -(C1-C4)alkyl[piperidinyl], -(C1-C4)alkyl[morpholinyl], -(C1-C4)alkyl[pyrrolidinyl], -(C1-C4)alkyl[diazazyl], -(C1-C4)alkyl[azacyclobutyl], piperazinyl, and tetrahydropyridinyl, wherein the piperidinyl, morpholinyl, pyrrolidinyl, diazazyl, tetrahydropyridinyl, azacyclobutyl, and each occurrence of piperazinyl are optionally selected from 1 to 3 of R 5a The group is substituted.

34. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 34, wherein R 5a Selected from (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' 1 .

35. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 34, wherein R 5a It 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.

36. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 35, wherein R c1 Selected from 5- to 7-membered heteroaryl and 4- to 6-membered heterocyclic groups, wherein each of the 5- to 7-membered heteroaryl and 4- to 6-membered heterocyclic groups is optionally composed of 1 to 3 groups selected from R 6a The group is substituted.

37. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 36, wherein R c1 It is piperidinyl or pyridinyl, each optionally composed of 1 to 3 groups selected from R 6a The group is substituted.

38. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 37, wherein R 6a It is a (C1-C4) alkyl group.

39. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 38, wherein R a1 and R b1 Each group is independently selected from hydrogen, -(C1-C4)alkyl [5- to 7-membered heteroaryl] and 4- to 6-membered heterocyclic groups, wherein for each occurrence of the 5- to 7-membered heteroaryl and the 4- to 6-membered heterocyclic group, each group is optionally composed of 1 to 3 groups selected from R 6a The group is substituted.

40. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 39, wherein R a1 and R b1 Each is independently selected from hydrogen and -(C1-C4)alkyl [5 to 7-membered heteroaryl], wherein the 5 to 7-membered heteroaryl group is optionally composed of 1 to 3 atoms selected from R 6a The group is substituted.

41. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 40, wherein R a1 It is hydrogen and R b1 It is -(C1-C4)alkyl[pyridinyl], wherein the pyridinyl group is optionally composed of 1 to 3 atoms selected from R 6a The group is substituted.

42. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 41, wherein R f1 It is optional to be selected from 1 to 3 R 8a The group substituted with a 4- to 6-membered heterocyclic group.

43. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 42, wherein R f1 It is optional to be selected from 1 to 3 R 8a The piperazine group is substituted with the group.

44. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 43, wherein R 8a It is a (C1-C4) alkyl group.

45. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 44, wherein R d1 and R e1 Each is independently selected from hydrogen, -(C1-C4)alkyl NR' 1 R'' 1 (C1-C4)alkyl [5- to 7-membered heteroaryl] and 4- to 6-membered heterocyclic groups, wherein the 5- to 7-membered heteroaryl and the 4- to 6-membered heterocyclic groups are each optionally composed of 1 to 3 groups selected from R 7a The group is substituted.

46. ​​The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 45, wherein R d1 and R e1 Each is independently selected from hydrogen, -(C1-C4)alkylN[(C1-C4)alkyl]2, (C1-C4)alkyl[pyridinyl] and piperidinyl, wherein the pyridinyl and piperidinyl are each optionally composed of 1 to 3 elements selected from R 7a The group is substituted.

47. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 46, wherein R g1 and R h1 Each is independently selected from 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 heterocyclic, wherein the phenyl, 5- to 7-membered heteroaryl and 4- to 6-membered heterocyclic are each optionally composed of 1 to 3 groups selected from R 7a The group is substituted.

48. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 47, wherein R g1 and R h1 Each of the following is independently selected from hydrogen, (C1-C4)alkyl, halo(C1-C4)alkyl, -(C1-C4)alkyl[pyridyl], -(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 the phenyl, pyridinyl, piperidinyl, pyrrolidinyl and morpholinyl are each optionally selected from 1 to 3 of R 7a The group is substituted.

49. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 48, wherein R 7a It is selected from (C1-C4)alkyl, halo(C1-C4)alkyl, halogen, (C1-C4)alkoxy and cyano.

50. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 49, wherein R 3 Selected from -CH2NHCH3, -CH2N(CH3)2, , -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, , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and .

51. The compound according to claim 1, wherein the compound is selected from... , , , , , , , , , , Or its pharmaceutically acceptable salt.

52. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 51; and a pharmaceutically acceptable carrier.

53. A method of treating a condition in a subject in need that responds to activation of the function of wild-type tumor suppressor protein TP53, comprising administering to the subject a therapeutically effective amount of the compound according to any one of claims 1 to 51, or a pharmaceutically acceptable salt thereof, or the composition according to claim 52.

54. The method of claim 53, wherein the condition is cancer.

55. The method of claim 53 or claim 54, wherein the condition is cancer carrying a Y220C mutation.

56. The method according to any one of claims 53 to 55, wherein the cancer is a solid tumor or a hematologic malignancy.

57. The method according to any one of claims 53 to 56, 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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