CROSS-REFERENCE TO RELATED APPLICATIONS FOR SMALL MOLECULE INHIBITORS OF KRAS MUTANT PROTEINS

Abstract

The compound of formula (I) or a pharma- ceutically acceptable salt thereof can inhibit the G12C, G12D and / or G12V mutants of the Kirsten Rat Sarcoma (KRAS) protein, and is expected to have utility as a therapeutic agent, for example, for treating cancer. The present disclosure also provides a pharmaceutical composition comprising the compound of formula (I) or a pharma- ceutically acceptable salt thereof. The present disclosure also relates to methods of using the compound or a pharma- ceutically acceptable salt thereof in the treatment and prevention of cancer, and methods of preparing medicaments therefor. [Formula 1] TIFF2024521979000129.tif60140

Description

[Technical Field]

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 194,852, filed May 28, 2021, which is incorporated herein by reference in its entirety. The present invention relates to small molecule inhibitors of Kirsten rat sarcoma (KRAS), for example, that inhibit the G12C, G12D, and G12V mutants of the KRAS protein, and to pharmaceutical compositions comprising compounds of formula (I) and methods of using such compounds for the treatment of diseases such as cancer. [Background technology]

[0002] RAS is a small, monomeric GTP-binding protein with a molecular mass of approximately 21 kDa that functions as a molecular on / off switch. RAS can bind GTP by binding to guanine nucleotide exchange factors (GEFs) (e.g., SOS1), which forces the release of the bound nucleotide and releases GDP. When RAS binds to GTP, it becomes activated (ON) and recruits and activates proteins necessary for signal propagation, such as c-Raf and PI3-kinase, from other receptors. RAS also possesses the enzymatic activity of cleaving the terminal phosphate of GTP nucleotides, converting them to GDP. The conversion rate is usually slow, but can be dramatically accelerated by proteins of the GTPase-activating protein (GAP) class, such as RasGAP. When GTP is converted to GDP, RAS is inactivated (OFF).

[0003] Commonly recognized members of the RAS subfamily include HRAS, KRAS, and NRAS. Among these, KRAS mutations have been observed in many malignancies, including 86% of pancreatic ductal adenocarcinomas (PDAC), 41% of colorectal cancers (CRC), and 32% of lung adenocarcinomas (LUAD; a subtype of non-small cell lung cancer (NSCLC)). Mutations frequently occur at the glycine residue at position 12 ("G12") of KRAS, with G12 mutations accounting for 91% (PDAC), 68% (CRC), and 85% (LUAD) of all KRAS mutations, respectively. The distribution of amino acid substitutions at G12 varies among tissue types. The most prevalent mutation in LUAD is a cysteine ​​("G12C") mutation (46%), whereas the predominant mutation in PDAC (45%) and CRC (45%) is a mutation to aspartic acid ("G12D"). Mutation of G12 to valine ("G12V") is observed in a significant proportion of G12 mutations in PDAC (35%), CRC (30%), and LUAD (23%) (Nature Reviews Drug Discovery, 19, 533-552, 2020). [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Nature Reviews Drug Discovery, 19, 533-552, 2020 Summary of the Invention

[0005] Intensive efforts are underway to develop KRAS-G12C inhibitors. Several covalent inhibitors focusing on cysteine ​​residues have been reported, and some of them, such as AMG510 (NCT03600883), MRTX849 (NCT03785249), and JNJ-74699157 (NCT04006301), are undergoing clinical studies. However, KRAS-G12C mutations represent only a small fraction of all KRAS mutations, primarily observed in LUAD. Effective inhibition of other commonly occurring KRAS mutant proteins, such as KRAS-G12D and KRAS-G12V, requires a different approach because these mutants lack a reactive cysteine ​​in the active site (Nature Reviews Drug Discovery, 19, 533-552, 2020).

[0006] The present disclosure provides small molecule inhibitors that modulate mutant KRAS, HRAS, and / or NRAS proteins and may be valuable pharmaceutically active compounds for the treatment of cancer. In some embodiments, the disclosed compounds selectively inhibit KRAS-G12C, KRAS-G12D, and / or KRAS-G12V proteins. Compounds of formula (I): [ka] and pharmaceutically acceptable salts thereof can modulate the activity of KRAS, HRAS, and / or NRAS, thereby affecting signal transduction pathways that regulate cell growth, differentiation, and proliferation associated with oncology disorders. In certain embodiments, compounds of formula (I) can inhibit KRAS-G12C, KRAS-G12D, and / or KRAS-G12V proteins. The present disclosure further provides methods for making compounds of formula (I), methods for using such compounds to treat oncology disorders, and pharmaceutical compositions comprising compounds of formula (I). DETAILED DESCRIPTION OF THE INVENTION

[0007] Disclosed Compounds In one embodiment, the present disclosure provides a compound having the structural formula (I) shown above, or a pharmaceutically acceptable salt thereof: During the ceremony, X is (i) 6- to 9-membered monocyclic, fused bicyclic, or bridged bicyclic heterocycloalkyl, wherein the heterocycloalkyl is saturated and contains 1 to 2 heteroatoms selected from the group consisting of N, S, and O; (ii) 8- to 10-membered spiroheterocycloalkyl, wherein the spiroheterocycloalkyl is saturated and contains 1 to 2 heteroatoms selected from the group consisting of N and O; (iii) [ka] and [ka] selected from the group consisting of: When X is (i) or (ii), X is unsubstituted or selected from the group consisting of halo, hydroxy, C1-C6 alkyl, C1-C3 hydroxyalkyl, C1-C6 fluoroalkyl, carboxy, carbamoyl, C1-C3 carboxyalkyl, oxo, cyano, cyanomethyl, amino, pyrazolyl, oxadiazolonyl, -NHC(O)C1-C3 alkoxyC1-C3 alkyl, -NHC(O)C1-C3 alkoxyC6-C 10 Aryl, C1-C3 alkoxy, methoxy(C1-C3)alkyl, amino(C1-C3)alkyl, C1-C3 alkylamino(C1-C3)alkyl, C1-C3 dialkylamino, C1-C3 dialkylamino(C1-C3)alkyl, and NHC(O)C5-C 10 1 to 4 R selected from the group consisting of heteroaryl X are independently substituted by substituents, and heteroaryl is optionally substituted by C1-C3 alkyl; Ring Y is a 9-10 membered bicyclic ring system, said ring system being partially unsaturated or aromatic, and Ring Y containing 0-2 nitrogen heteroatoms; Ring Y is unsubstituted or contains 1 to 4 R selected from the group consisting of halo, hydroxy, amino, C1-C3 alkyl, C2-C3 alkynyl, and C1-C3 fluoroalkyl. y are independently substituted by substituents; Z is (i) a 5-8 membered monocyclic or bicyclic heterocycloalkyl, wherein the heterocycloalkyl is saturated and contains one nitrogen heteroatom, and the heterocycloalkyl is unsubstituted or contains one substituent R selected from the group consisting of halo, C-C alkyl, and methylene(C-C alkyl)(C-C alkyl)carbamate; ZHC is replaced by ]; (ii) [ka] wherein M is selected from the group consisting of hydroxy, C1-C3 dialkylamino, and C1-C4 alkylamino, and said cyclopropyl group is unsubstituted or substituted with up to two halo groups; (iii) [ka] wherein P is a 5-8 membered monocyclic, fused bicyclic, or bridged bicyclic heterocycloalkyl, wherein the heterocycloalkyl is saturated and contains 1-2 heteroatoms selected from the group consisting of N and O, and the heterocycloalkyl is unsubstituted or substituted with one R selected from the group consisting of halo, hydroxy, C1-C3 hydroxyalkyl, C1-C3 cyanoalkyl, carbamoyl, C1-C3 alkoxy, cyano, -NHC(O)C1-C3 alkyl, and oxadiazolonyl. P and the cyclopropyl group is unsubstituted or substituted with up to two halo groups. selected from the group consisting of: The subscript m is 0 or 1; The subscript n is 1 or 2.

[0008] In another embodiment, the present disclosure provides a method in which ring Y is [ka] The present invention provides a compound of formula (I),

[0009] In another embodiment, the present disclosure provides a method in which ring Y is [ka] The present invention provides a compound of formula (I),

[0010] In another embodiment, the present disclosure provides a method for treating a cancer, wherein X is [ka] and X is unsubstituted or 1 to 4 R X Compounds of formula (I) are provided, each independently substituted by a substituent.

[0011] In another embodiment, the present disclosure provides an aryl group selected from the group consisting of halo, hydroxy, C1-C6 alkyl, C1-C3 hydroxyalkyl, C1-C6 fluoroalkyl, carboxy, carbamoyl, C1-C3 carboxyalkyl, oxo, cyano, cyanomethyl, amino, pyrazolyl, oxadiazolonyl, —NHC(O)C1-C3 alkoxyC1-C3 alkyl, —NHC(O)C1-C3 alkoxyC6-C 10 Aryl and NHC(O)C5-C 10 1 to 4 R selected from the group consisting of heteroaryl X The present invention provides a compound of formula (I) substituted by a substituent, wherein the heteroaryl is optionally substituted by C1-C3 alkyl.

[0012] In another embodiment, the present disclosure provides a method for treating a cancer, wherein X is [ka] TIFF2024521979000011.tif193153TIFF2024521979000012.tif186151TIFF2024521979000013.tif82150.

[0013] In another embodiment, the present disclosure provides a method for treating a cancer, wherein X is [ka] and the subscript p is 0, 1 or 2.

[0014] In another embodiment, the present disclosure provides a method for treating a cancer, wherein X is [ka] The present invention provides a compound of formula (I) wherein

[0015] In another embodiment, the present disclosure provides a method for treating a cancer, wherein Z is [ka] TIFF2024521979000017.tif224149TIFF2024521979000018.tif205149 provides a compound of formula (I).

[0016] In another embodiment, the disclosure provides compounds of formula (I) wherein subscript m is 1.

[0017] In another embodiment, the disclosure provides compounds of formula (I) wherein the subscript n is 1.

[0018] In another embodiment, the disclosure provides compounds of formula (I) wherein the subscript n is 2.

[0019] In specific embodiments, the present disclosure provides a compound or a pharmaceutically acceptable salt thereof according to any one of Examples 1 to 151 set forth below.

[0020] The present disclosure includes pharmaceutically acceptable salts of the compounds defined herein, including pharmaceutically acceptable salts of all structural formulas, embodiments and classes defined herein.

[0021] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0022] As used throughout this disclosure, "a compound of Formula (I)" should be understood to encompass "a compound of Formula (I) or a pharmaceutically acceptable salt thereof." Similarly, "a compound of Formula (I)," "a compound disclosed herein," "a compound described herein," "a compound of the disclosure," and the like are used interchangeably and include both the compound, as well as a pharmaceutically acceptable salt thereof.

[0023] "Alkyl," as well as other groups having the prefix "alk," such as alkoxy, refer to carbon chains which may be linear or branched, or combinations thereof, containing the designated number of carbon atoms. For example, C1-C6 alkyl refers to an alkyl group having from 1 (i.e., methyl) up to 6 carbon atoms (i.e., hexyl). In certain embodiments, linear alkyl groups have from 1 to 6 carbon atoms, and branched alkyl groups have from 3 to 7 carbon atoms. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec- and tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl, and the like.

[0024] "Alkoxy" and "alkyl-O-" are used interchangeably and refer to an alkyl group linked to oxygen.

[0025] "Alkoxyalkyl" means an alkoxy-alkyl group in which the alkoxy and alkyl groups are as previously defined. The bond to the parent moiety is through a carbon atom of the alkyl component. Non-limiting examples of suitable alkoxyalkyl groups include methoxyalkyl groups, such as methoxymethyl and methoxyethyl.

[0026] "Alkynyl" means an aliphatic hydrocarbon group containing at least one carbon-carbon triple bond and which may be straight-chained or branched. Non-limiting examples include ethynyl, propynyl, and butynyl.

[0027] "Aryl" means a monocyclic, bicyclic, or tricyclic carbocyclic aromatic ring or ring system containing from 5 to 14 carbon atoms and in which at least one ring is aromatic. Non-limiting examples include phenyl and naphthyl.

[0028] "Alkoxyaryl" means an alkoxy-aryl group in which the alkoxy and aryl groups are as previously defined. The bond to the parent moiety is through a carbon atom of the aryl component. Non-limiting examples of suitable alkoxyaryl groups include methoxyphenyl.

[0029] "Aminoalkyl" means an -alkyl-NH group, where alkyl is as defined above. The bond to the parent moiety is through a carbon atom of the alkyl component. Non-limiting examples of suitable aminoalkyl groups include aminomethyl and aminoethyl. "Alkylamino" means an -NH-alkyl group, where alkyl is as defined above. The bond to the parent moiety is through the nitrogen of the amino component.

[0030] "Bicyclic ring system" refers to two linked rings. The rings may be fused, i.e., sharing two adjacent atoms, or "spirocyclic," i.e., sharing only a single atom, or "bridged," i.e., sharing three or more atoms, with two bridgehead atoms connected by a bridge containing at least one atom. Similarly, bicyclic rings may be aryl rings, heterocyclic rings, cycloalkyl rings, etc.

[0031] "Carbamoyl" refers to the monovalent group HN-C(O)- formed by loss of the -OH group of a carbamic acid. The bond to the parent group is through the carbon atom of the carbonyl moiety.

[0032] "Carboxyalkyl" means a carboxy(COOH)-alkyl group, wherein the alkyl group is as defined above. The bond to the parent group is through a carbon atom of the alkyl moiety.

[0033] "Cyanoalkyl" means an -alkyl-CN group, where alkyl is as previously defined. The bond to the parent moiety is through a carbon atom of the alkyl component. Non-limiting examples of suitable cyanoalkyl groups include cyanomethyl and 3-cyanopropyl.

[0034] "Cycloalkyl" refers to a saturated cyclic hydrocarbon group. In certain embodiments, a cycloalkyl group has 3 to 12 carbon atoms, forming 1 to 3 fused carbon rings. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, and the like.

[0035] "Dialkylamino" means an alkylamino as defined above wherein the amino atom is substituted by two alkyl substituents, which may be the same or different, e.g., -N(CH) or -N(CH)(CHCH).

[0036] "Dialkylaminoalkyl" means an aminoalkyl, as defined above, where the amino atom is also substituted with two alkyl substituents. The alkyl groups substituted on the amino atom can be the same or different. Non-limiting examples of suitable dialkylaminoalkyl groups include dimethylaminomethyl [(CH)NCH-] and N-ethyl-N-methylaminoethyl [(CHCH)(CH)N-CHCH-].

[0037] "Fluoroalkyl" includes mono- as well as multiply fluoro-substituted alkyl groups up to perfluoro-substituted alkyl, such as fluoromethyl, 1,1-difluoroethyl, trifluoromethyl, or 1,1,1,2,2-pentafluorobutyl.

[0038] "Halogen" or "halo," unless otherwise specified, includes fluorine (fluoro), chlorine (chloro), bromine (bromo), and iodine (iodo). In one embodiment, halo is fluoro (-F) or chloro (-Cl).

[0039] "Heteroaryl" refers to aromatic monocyclic and bicyclic ring systems in which one or more atoms in the ring is a heteroatom(s) that is an element other than carbon. "Heteroaryl" refers to aromatic monocyclic, bicyclic, and tricyclic ring systems in which one or more atoms in the ring, the heteroatom, is an element other than carbon. Heteroatoms are typically O, S, or N atoms. Examples of heteroaryl groups include pyrazolyl, oxadiazolonyl, pyridinyl, pyrimidinyl, pyrrolyl, pyridazinyl, isoxazolyl, thiazolyl, oxazolyl, indolyl, benzoxazolyl, benzothiazolyl, and imidazolyl.

[0040] "Heterocycloalkyl" or "heterocyclic ring" or "heterocycle" means a non-aromatic monocyclic, bicyclic, tricyclic, or bridged ring system containing about 3 to about 10 ring atoms, preferably about 5 to about 10 ring atoms, in which one or more atoms in the ring system is an element other than carbon, for example, nitrogen, oxygen, phosphorus, or sulfur, alone or in combination. There are no adjacent oxygen and / or sulfur atoms in the ring system. In some embodiments, a heterocycloalkyl contains about 5 to about 6 ring atoms. The prefix aza, oxa, phospha, or thia before the heterocycle root name means that at least a nitrogen, oxygen, phosphorus, or sulfur atom, respectively, is present as a ring atom. In some embodiments, the nitrogen or sulfur atom of a heterocycloalkyl can be optionally oxidized to the corresponding N-oxide, S-oxide, or S,S-dioxide. Non-limiting examples of suitable monocyclic heterocycles include piperidyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, 1,4-dioxanyl, tetrahydrofuranyl, tetrahydrothiophenyl, phosphorinane, phosphinane, 1-oxophosphinan-1-ium, etc. "Spiroheterocycloalkyl" refers to a fused ring system in which the rings share only a single atom and at least one of the rings is a heterocycloalkyl.

[0041] "Hydroxyalkyl" means an HO-alkyl- group, where alkyl is as defined above. The bond to the parent moiety is through a carbon atom of the alkyl group. Preferred hydroxyalkyl groups contain lower alkyl. Non-limiting examples of suitable hydroxyalkyl groups include hydroxymethyl and 2-hydroxyethyl.

[0042] "Methylene(C1-C3 alkyl)(C1-C3 alkyl)carbamate" has the following structure: [ka] In other words, a carbamate group has an alkyl group, as defined above, attached to the nitrogen atom.

[0043] Any variable part (e.g., R y When R occurs more than once in any constituent or in Formula (I) or other general formulas herein, its definition at each occurrence is independent of its definition at every other occurrence. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds. In selecting compounds of the present disclosure, one of skill in the art will recognize various substituents, i.e., R, that are consistent with well-known principles of connectivity and stability of chemical structures. y It will be recognized that the substituents should be selected from the group consisting of aryl, heteroaryl, and saturated heteroaryl. Unless expressly stated otherwise, substitution by the specified substituents is permissible at any atom within the ring (e.g., an aryl, heteroaryl, or saturated heteroaryl ring), provided that such ring substitution is chemically feasible and results in a stable compound. A "stable" compound is one that can be prepared and isolated and whose structure and properties remain essentially unchanged, or can be made to remain essentially unchanged, for a period of time sufficient to permit use of the compound for the purposes described herein (e.g., therapeutic or prophylactic administration to a subject).

[0044] The term "substituted" should be considered to include multiple degrees of substitution with a specified substituent. Where multiple substituent moieties are disclosed or claimed, the substituted compound may be independently substituted one or more times with one or more of the disclosed or claimed substituent moieties. By independently substituted, it is intended that the (two or more) substituents may be the same or different.

[0045] Unless expressly shown or described otherwise, variables having "fluid" bonds depicted in structural formulae, e.g., R Xis allowed at any available carbon atom in the ring to which the variable is attached. When a moiety is described in Formula (I) or any embodiment thereof as "optionally substituted," this means that Formula (I) or any embodiment thereof encompasses compounds that do not contain the substituent(s) described on the moiety, as well as compounds that contain the substituent(s) described on the moiety.

[0046] Wavy line [ka] As used herein, indicates the point of attachment to the remainder of the compound.

[0047] The compounds of formula (I) may contain one or more asymmetric centers and may therefore exist as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures, and individual diastereomers. All asymmetric centers present in the compounds of formula (I) are independent of one another and may have either the S or R configuration. The compounds of formula (I) encompass all possible enantiomers and diastereomers, as well as mixtures of two or more stereoisomers in any ratio, for example, mixtures of enantiomers and / or diastereomers. Thus, enantiomers in enantiomerically pure form, both as levorotatory and dextrorotatory antipodes, in the form of racemates, and in the form of mixtures of the two enantiomers in any ratio, are the subject of the present disclosure. In the case of cis / trans isomerism, the present disclosure encompasses both the cis and trans forms and mixtures of these forms in any ratio. The present disclosure is intended to encompass all such stereoisomeric forms of the compounds of formula (I). When a structural formula or chemical name specifies a specific configuration at a stereocenter, the enantiomer or stereoisomer of the compound resulting from the specified stereocenter is intended. When a straight line is shown at a chiral center in the structural formula of a compound of formula (I), the structural formula encompasses both the S and R stereoisomers associated with that chiral center, as well as mixtures thereof.

[0048] The compounds of formula (I) can be separated into their individual diastereoisomers, for example, by fractional crystallization with suitable solvents, such as methanol or ethyl acetate, or mixtures thereof, or by chiral chromatography using an optically active stationary phase. Absolute stereochemistry can be determined by X-ray crystallography of crystalline products or crystalline intermediates which are derivatized, if necessary, with a reagent containing an asymmetric center of known absolute configuration. Vibrational circular dichroism (VCD) can also be used to determine absolute stereochemistry. Alternatively, any stereoisomer or isomers of the compounds of formula (I) can be obtained by stereospecific synthesis using optically pure starting materials or reagents of known absolute configuration.

[0049] If desired, the racemic mixture of the compound can be separated so that the individual enantiomers are isolated. Separation can be carried out by methods well known in the art, for example, by coupling the racemic mixture of the compound with an enantiomerically pure compound to form a diastereomeric mixture, followed by standard methods, such as fractional crystallization or chromatography, to separate the individual diastereomers. The coupling reaction is often the formation of a salt with an enantiomerically pure acid or base. The diasteromeric derivative can then be converted to the pure enantiomer by cleavage of the added chiral residue. Alternatively, the racemic mixture of the compound can be directly separated by chromatographic methods using a chiral stationary phase, which methods are well known in the art.

[0050] For compounds of Formula (I) containing olefinic double bonds, unless otherwise specified, both E and Z geometric isomers are intended to be encompassed.

[0051] Some compounds described herein may exist as tautomers, with different points of attachment of hydrogen and shifts in one or more double bonds. For example, a ketone and its enol form are keto-enol tautomers. Individual tautomers as well as mixtures thereof are encompassed by the compounds of formula (I).

[0052] Some compounds of Formula (I) described herein may exist as atropisomers when the rotational energy barrier around a single bond is high enough to prevent free rotation at a given temperature, thus allowing for the isolation of individual atropisomers with different properties. Individual atropisomers as well as mixtures thereof are encompassed by the compounds of Formula (I) of the present disclosure. Once resolved, individual atropisomers can be designated by established conventions, such as those set forth in the International Union of Pure and Applied Chemistry (IUPAC) 2013 Recommendations.

[0053] In compounds of formula (I), atoms may be present at their natural isotopic abundance, or one or more atoms may be artificially enriched in a particular isotope having the same atomic number but an atomic mass or mass number different from that found predominantly in nature. The present disclosure, as described and claimed herein, is intended to encompass all suitable isotopic variations of compounds of formula (I) and embodiments thereof. For example, different isotopic forms of hydrogen (H) include protium ( 1 H) and deuterium ( 2 H, also referred to herein as D). Protium is the predominant hydrogen isotope found in nature. Enrichment with deuterium may provide certain therapeutic benefits, such as increased in vivo half-life or reduced dosage requirements, or may result in compounds useful as standards for characterization of biological samples. Isotopically enriched compounds of formula (I) can be prepared without undue experimentation by conventional methods well known to those skilled in the art, or by methods similar to those described in the schemes and examples herein, using appropriate isotopically enriched reagents and / or intermediates.

[0054] The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids. When the compound of formula (I) is acidic, its corresponding salt can be conveniently prepared from pharmaceutically acceptable non-toxic bases, such as inorganic and organic bases. Salts derived from such inorganic bases include aluminum, ammonium, calcium, copper (both ferrous and ferrous), ferric, ferrous, lithium, magnesium, manganese (both ferrous and ferrous), potassium, sodium, zinc, and the like salts. Preferred are ammonium, calcium, magnesium, potassium, and sodium salts. Salts prepared from pharmaceutically acceptable non-toxic organic bases include salts of primary, secondary, and tertiary amines derived from both naturally occurring and synthetic sources. Examples of pharmaceutically acceptable organic non-toxic bases that can form salts include, for example, arginine, betaine, caffeine, choline, N,N ’ -dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethyl-morpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, cyclohexylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like.

[0055] When the compound of formula (I) is basic, its corresponding salt can be easily prepared from pharmaceutically acceptable non-toxic inorganic and organic acids. Examples of such acids include acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, etc. Preferred are citric acid, hydrobromic acid, hydrochloric acid, maleic acid, phosphoric acid, sulfuric acid, and tartaric acid. When the compound of formula (I) contains both an acidic and a basic group in the molecule, the present disclosure also encompasses internal salts or betaines (zwitterions) in addition to the salt forms described above. Salts can be obtained from compounds of formula (I) by conventional methods known to those skilled in the art, for example by combining with organic or inorganic acids or bases in a solvent or dispersant, or by anion or cation exchange with other salts. The present disclosure also encompasses any salts of compounds of formula (I) that are not suitable for direct use in pharmaceuticals due to low physiological compatibility, but can be used, for example, as intermediates for chemical reactions or for the preparation of pharmaceutically acceptable salts.

[0056] Additionally, compounds of formula (I) may exist in amorphous form and / or one or more crystalline forms, and as such, all amorphous and crystalline forms of compounds of formula (I), including the present examples, and mixtures thereof, are intended to be encompassed within the scope of this disclosure. Additionally, some compounds of formula (I) may form solvates with water (i.e., hydrates) or common organic solvents, such as, but not limited to, ethyl acetate. Such solvates and hydrates of the compounds of the present invention, particularly pharmaceutically acceptable solvates and hydrates, are also encompassed within the scope of this disclosure, along with unsolvated and anhydrous forms.

[0057] Also within the scope of this disclosure are any pharmaceutically acceptable prodrug modifications of compounds of formula (I) that result in conversion in vivo to compounds within the scope of this disclosure.

[0058] The term "therapeutically effective (or efficacious) amount" and similar descriptions, such as "amount effective for treatment" or "effective dose," are intended to mean an amount of a compound of Formula (I) that elicits the biological or medical response in a tissue, system, animal, or human that a researcher, veterinarian, physician, or other clinical practitioner is seeking. In a preferred embodiment, the term "therapeutically effective amount" refers to an amount of a compound of Formula (I) that alleviates at least one clinical symptom in a human patient. The term "prophylactically effective (or efficacious) amount" and similar descriptions, such as "prophylactically effective amount," are intended to mean an amount of a compound of Formula (I) that inhibits or reduces the risk of occurrence of the biological or medical event that a researcher, veterinarian, physician, or other clinical practitioner is seeking to prevent in a tissue, system, animal, or human.

[0059] Dosage of Compounds of Formula (I) The administration method using the compound of formula (I) is selected according to various factors, such as the type, species, age, weight, sex, and medical condition of the patient; the severity of the condition to be treated; the efficacy of the compound selected for administration; the route of administration; and the patient's renal and hepatic function. It is well within the ordinary skill of a skilled clinician to consider such factors in order to determine the therapeutically effective or prophylactically effective dosage required to prevent, inhibit, or halt the progression of the condition. It is understood that a specific daily dosage may simultaneously be both a therapeutically effective amount for treating, for example, a neoplastic condition, and a prophylactically effective amount for preventing, for example, a neoplastic condition.

[0060] While individual needs vary, determining the optimal range of effective amounts of a compound of formula (I) is within the skill of the art. For example, for administration to humans in the curative or prophylactic treatment of the conditions and disorders identified herein, a typical dosage of a compound of formula (I) can be about 0.05 mg / kg / day to about 50 mg / kg / day, or at least 0.05 mg / kg, or at least 0.08 mg / kg, or at least 0.1 mg / kg, or at least 0.2 mg / kg, or at least 0.3 mg / kg, or at least 0.4 mg / kg, or at least 0.5 mg / kg, and any amount therebetween, to about 50 mg / kg or less, or about 40 mg / kg or less, or about 30 mg / kg or less, or about 20 mg / kg or less, or about 10 mg / kg or less, and any value therebetween, such as about 2.5 mg / day (0.5 mg / kg x 5 kg) to about 5000 mg / day (50 mg / kg x 100 kg). For example, the dosage of the compound may be from about 0.1 mg / kg / day to about 50 mg / kg / day, or from about 0.05 mg / kg / day to about 10 mg / kg / day, or from about 0.05 mg / kg / day to about 5 mg / kg / day, or from about 0.05 mg / kg / day to about 3 mg / kg / day, or from about 0.07 mg / kg / day to about 3 mg / kg / day, or from about 0.09 mg / kg / day to about 3 mg / kg / day, or from about 0.05 mg / kg / day to about 0.1 mg / kg / day, Alternatively, it may be about 0.1 mg / kg / day to about 1 mg / kg / day, or about 1 mg / kg / day to about 10 mg / kg / day, or about 1 mg / kg / day to about 5 mg / kg / day, or about 1 mg / kg / day to about 3 mg / kg / day, or about 3 mg / day to about 500 mg / day, or about 5 mg / day to about 250 mg / day, or about 10 mg / day to about 100 mg / day, or about 3 mg / day to about 10 mg / day, or about 100 mg / day to about 250 mg / day. Such a dose may be administered in a single dose or divided into multiple doses.

[0061] Pharmaceutical Composition The compounds of formula (I) and their pharmaceutically acceptable salts can be administered to animals, preferably mammals, particularly humans, as pharmaceuticals, either alone, in admixture with one another, or in the form of pharmaceutical compositions. The term "subject" or "patient" includes animals, preferably mammals, particularly humans, who use the active agents of the present invention for the prevention or treatment of a medical condition. Administration of the drug to a subject includes both self-administration and administration to the patient by another. The subject may be a subject who needs or desires treatment for an existing disease or medical condition, or a subject who needs or desires prophylactic treatment to prevent or reduce the risk of developing the disease or medical condition. As used herein, a subject "in need of" treatment or prophylactic treatment for an existing condition includes both a medical professional's determination of the need and the patient's desire for such treatment.

[0062] Accordingly, the present disclosure also provides compounds of formula (I) and pharmaceutically acceptable salts thereof for use as pharmaceuticals, their use for modulating the activity of mutant KRAS, HRAS and / or NRAS proteins, in particular their use in the treatment and prevention of the diseases or disorders described below, and their use for preparing medicaments for such purposes. In certain embodiments, compounds of formula (I) and pharmaceutically acceptable salts thereof inhibit KRAS G12C, KRAS-G12D and / or KRAS-G12V proteins.

[0063] The present disclosure further provides pharmaceutical compositions comprising an effective dose of at least one compound of formula (I) and / or a pharmaceutically acceptable salt thereof as an active ingredient, together with a conventional pharmaceutically acceptable carrier, i.e., one or more pharmaceutically acceptable carrier substances and / or excipients.

[0064] Thus, the present disclosure provides, for example, at least one compound of formula (I) and / or a pharmaceutically acceptable salt thereof for use as a pharmaceutical composition comprising an effective dose of said compound as an active ingredient and a conventional pharmaceutically acceptable carrier, and the use of said compound and / or a pharmaceutically acceptable salt thereof in the treatment or prevention of the diseases or disorders described below, for example cancer, and its use for preparing a medicament for such purposes.

[0065] The pharmaceutical compositions according to the present disclosure can be administered orally, for example, in the form of pills, tablets, lacquered tablets, dragees, granules, hard and soft gelatin capsules, aqueous, alcoholic or oily solutions, syrups, emulsions or suspensions, or rectally, for example, in the form of suppositories. Administration can also be parenteral, for example, subcutaneously, intramuscularly or intravenously in the form of solutions for injection or infusion.

[0066] Other suitable administration forms are, for example, ointments, tinctures, sprays or transdermal therapeutic systems or transdermal or topical administration, for example in the form of microcapsules, implants or rods. The preferred administration form depends, for example, on the disease to be treated and its severity.

[0067] The amount of the active compound of the compound described herein and / or a pharmaceutically acceptable salt thereof in the pharmaceutical composition is typically 0.01 to 200 mg, or 0.1 to 200 mg, or 1 to 200 mg per dose, but may be higher depending on the type of pharmaceutical composition. In some embodiments, the amount of the active compound of the compound of Formula (I) and / or a pharmaceutically acceptable salt thereof in the pharmaceutical composition is 0.01 to 10 mg per dose. The pharmaceutical composition typically contains 0.5 to 90 weight percent of at least one compound of Formula (I) and / or a pharmaceutically acceptable salt thereof. The preparation of pharmaceutical compositions can be carried out in a manner known per se. For this purpose, one or more compounds of Formula (I) and / or a pharmaceutically acceptable salt thereof, together with one or more solid or liquid pharmaceutical carrier materials and / or additives (or auxiliary substances), optionally in combination with other pharmaceutically active compounds having a therapeutic or prophylactic action, are formed into a suitable administration form or dosage form, which can then be used as a medicament in human or veterinary medicine.

[0068] For the preparation of pills, tablets, dragees, and hard gelatin capsules, lactose, starch (e.g., corn starch or starch derivatives), talc, stearic acid, or its salts, can be used. Carriers for soft gelatin capsules and suppositories include fats, waxes, semi-solid and liquid polyols, natural or hardened oils, etc. Suitable carriers for the preparation of liquids, such as injectable solutions, emulsions, or syrups, include, for example, water, physiologically acceptable sodium chloride solution, alcohols, such as ethanol, glycerol, polyols, sucrose, invert sugar, glucose, mannitol, vegetable oils, etc. It is also possible to lyophilize the compound of formula (I) and its pharmaceutically acceptable salts, and use the resulting lyophilized product, for example, to prepare preparations for injection or infusion. Suitable carriers for microcapsules, implants, or rods include, for example, glycolic acid and lactic acid copolymers.

[0069] In addition to the active compound and carrier, the pharmaceutical composition may also contain conventional additives such as fillers, disintegrants, binders, lubricants, wetting agents, stabilizers, emulsifiers, dispersing agents, preservatives, sweeteners, colorants, flavors, fragrances, thickeners, diluents, buffer substances, solvents, solubilizers, agents for obtaining a depot effect, salts for modifying osmotic pressure, coating agents and / or antioxidants.

[0070] Methods of Use of Compounds of Formula (I) The present application provides a method for inhibiting RAS-mediated cell signaling, comprising contacting a cell with a compound of formula (I) or a pharmaceutically acceptable salt thereof. Inhibition of RAS-mediated signaling can be assessed and demonstrated by a variety of methods known in the art. Non-limiting examples include: (a) a decrease in the GTPase activity of RAS; (b) a decrease in GTP binding affinity or an increase in GDP binding affinity; (c) a decrease in the K off Increase in or GDP K off (d) reduced levels of downstream signaling molecules in the RAS pathway, such as reduced levels of pMEK, pERK, or pAKT; and / or (e) reduced binding of the RAS complex to downstream signaling molecules, such as, but not limited to, Raf. Kits and commercially available assays are available to measure one or more of the above.

[0071] The present application also provides methods of using a compound of formula (I) (or a pharmaceutically acceptable salt thereof) or a pharmaceutical composition comprising such a compound to treat disease states, including but not limited to conditions involving mutant KRAS, HRAS and / or NRAS proteins (e.g., cancer), and in some embodiments conditions involving KRAS G12C, KRAS-G12D and / or KRAS-G12V mutants.

[0072] In some embodiments, a method for treating cancer is provided, the method comprising administering a therapeutically effective amount of a compound of Formula (I) (or a pharmaceutically acceptable salt thereof) or any of the aforementioned pharmaceutical compositions containing such a compound to a subject in need of such treatment. In some embodiments, the cancer is mediated by a KRAS, HRAS, or NRAS mutation, such as a KRAS G12C, KRAS-G12D, and / or KRAS-G12V mutation. In various embodiments, the cancer is pancreatic cancer, colorectal cancer, or lung cancer. In some embodiments, the cancer is gallbladder cancer, thyroid cancer, or bile duct cancer.

[0073] In some embodiments, the disclosure provides a method of treating a disorder in a subject in need thereof, the method comprising determining whether the subject has a KRAS, HRAS, or NRAS mutation (e.g., a KRAS G12C, KRAS-G12D, and / or KRAS-G12V mutation), and if the subject is determined to have the KRAS, HRAS, or NRAS mutation, administering to the subject a therapeutically effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

[0074] The compounds of the present disclosure have the potential to inhibit anchorage-independent cell growth and therefore inhibit tumor metastasis. Accordingly, another embodiment of the present disclosure provides a method for inhibiting tumor metastasis, comprising administering an effective amount of a compound of formula (I).

[0075] Mutations in KRAS, HRAS, or NRAS have also been identified in hematopoietic malignancies (e.g., cancers affecting the blood, bone marrow, and / or lymph nodes). Accordingly, certain embodiments relate to administering a compound of Formula (I) (e.g., in the form of a pharmaceutical composition) to a subject in need of treatment for a hematopoietic malignancy. Such malignancies include, but are not limited to, leukemia and lymphoma. For example, compounds of the present disclosure can be used to treat diseases such as acute lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), chronic myelogenous leukemia (CML), acute monocytic leukemia (AMoL), and / or other leukemias. In other embodiments, the compounds are useful for treating lymphomas, such as Hodgkin's lymphoma or non-Hodgkin's lymphoma. In various embodiments, the compounds are useful in the treatment of plasma cell malignancies, such as multiple myeloma, mantle cell lymphoma, and Waldenstrom's macroglobulinemia.

[0076] Determining whether a tumor or cancer contains a KRAS, HRAS, or NRAS mutation (e.g., a KRAS G12C, KRAS-G12D, and / or KRAS-G12V mutation) can be done by assessing the nucleotide sequence encoding the KRAS, HRAS, or NRAS protein, by assessing the amino acid sequence of the KRAS, HRAS, or NRAS protein, or by assessing the characteristics of a putative KRAS, HRAS, or NRAS mutant protein. Wild-type human KRAS, HRAS, or NRAS sequences are known in the art.

[0077] Methods for detecting mutations in the nucleotide sequence of KRAS, HRAS, or NRAS are also known to those skilled in the art. Such methods include, but are not limited to, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay, polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) assay, real-time PCR assay, PCR sequencing, mutant allele-specific PCR amplification (MASA) assay, direct sequencing, primer extension reaction, electrophoresis, oligonucleotide ligation assay, hybridization assay, TaqMan assay, SNP genotyping assay, high-resolution melting curve assay, and microarray analysis. In some embodiments, a sample is evaluated for KRAS, HRAS, or NRAS mutations (e.g., KRAS G12C, KRAS-G12D, and / or KRAS-G12V mutations) by real-time PCR. In real-time PCR, a fluorescent probe specific to the KRAS, HRAS, or NRAS mutation is used. If a mutation is present, the probe binds and fluorescence is detected. In some embodiments, the KRAS, HRAS, or NRAS mutation is identified using direct sequencing of specific regions within the KRAS, HRAS, or NRAS gene (eg, exon 2 and / or exon 3).

[0078] Methods for detecting mutations in KRAS, HRAS, or NRAS proteins (e.g., KRAS G12C, KRAS-G12D, and / or KRAS-G12V mutations) are known to those skilled in the art, including, but not limited to, detection of mutant KRAS, HRAS, or NRAS proteins using binding agents (e.g., antibodies) specific for the mutant forms of the protein, protein electrophoresis and Western blotting, and direct peptide sequencing.

[0079] Many tissue samples can be evaluated to determine whether a tumor or cancer contains a KRAS, HRAS, or NRAS mutation (e.g., a KRAS G12C, KRAS-G12D, and / or KRAS-G12V mutation). In some embodiments, the sample is collected from a subject with a tumor or cancer. In some embodiments, the sample is a freshly collected tumor / cancer sample. In some embodiments, the sample is a frozen tumor / cancer sample. In some embodiments, the sample is a formalin-fixed, paraffin-embedded sample. In some embodiments, the sample is a circulating tumor cell (CTC) sample. In some embodiments, the sample is processed into a cell lysate. In some embodiments, the sample is processed into DNA or RNA.

[0080] The present application also provides a method of treating a hyperproliferative disorder, comprising administering a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof. In some embodiments, the method is directed to treating cancer, such as acute myeloid leukemia, adolescent cancer, childhood adrenocortical carcinoma, AIDS-related cancers (e.g., lymphoma and Kaposi's sarcoma), anal cancer, appendix cancer, astrocytoma, atypical teratoid, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain stem glioma, brain tumor, breast cancer, bronchial tumor, Burkitt's lymphoma, carcinoid tumor, atypical teratoid, embryonal tumor, germ cell tumor, primary lymphoma, cervical cancer, childhood cancer, chordoma, cardiac tumor, chronic lymphocytic leukemia, and the like. Hematologic malignancies include: chronic myeloid leukemia (CLL), chronic myelogenous leukemia (CML), chronic myeloproliferative disorders, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic ductal carcinoma in situ (DCIS), germinoma, CNS cancer, endometrial cancer, ependymoma, esophageal cancer, nasal neuroblastoma, Ewing's sarcoma, extracranial germ cell tumor, extragonadal germ cell tumor, eye cancer, fibrous histiocytoma of bone, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic tumor, hairy cell leukemia, head and neck cancer, Cancer of the heart, liver cancer, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, pancreatic islet cell tumor, pancreatic neuroendocrine tumor, kidney cancer, laryngeal cancer, lip and oral cavity cancer, liver cancer, lobular carcinoma in situ (LCIS), lung cancer, lymphoma, metastatic squamous cell carcinoma of the neck of unknown primary, midline carcinoma, oral cancer; multiple endocrine neoplasia syndrome, multiple myeloma / plasma cell neoplasm, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, multiple myeloma, Merkel cell carcinoma, malignant mesothelioma, malignant fibrous histiocytoma of bone, and Osteosarcoma, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer (NSCLC), oral cavity cancer, lip and oral cavity cancer, oropharyngeal cancer, ovarian cancer, pancreatic cancer, papillomatosis, paraganglioma, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pleuropulmonary blastoma, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, transitional cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, stomach (gastric) cancer, small cell lung cancer;The present invention relates to the treatment of subjects suffering from small intestine cancer, soft tissue sarcoma, T-cell lymphoma, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, trophoblastic tumor, unusual cancer of childhood, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, or virally induced cancer. In some embodiments, the method relates to the treatment of non-cancerous hyperproliferative disorders, such as benign thickening of the skin (e.g., psoriasis), restenosis, or prostate (e.g., benign prostatic hyperplasia (BPH));

[0081] In some embodiments, the method for treatment relates to the treatment of lung cancer, and the method comprises administering a therapeutically effective amount of a compound of formula (I) (or a pharmaceutical composition comprising such a compound) to a subject in need thereof. In some particular embodiments, the lung cancer is non-small cell lung cancer (NSCLC), such as adenocarcinoma, squamous cell lung carcinoma or large cell lung carcinoma. In some embodiments, the lung cancer is small cell lung carcinoma. Other lung cancers that can be therapeutically beneficial by the compound of formula (I) include, but are not limited to, adenocarcinoma, carcinoid tumor and undifferentiated carcinoma.

[0082] The present disclosure also provides methods for modulating the activity of a mutant KRAS, HRAS, or NRAS protein (e.g., activity resulting from KRAS G12C, KRAS-G12D, and / or KRAS-G12V mutations) by contacting the protein with an effective amount of a compound of Formula (I). Modulation can be inhibition or activation of the protein's activity. In some embodiments, the present disclosure provides methods for inhibiting the activity of a mutant KRAS, HRAS, or NRAS protein (e.g., KRAS G12C, KRAS-G12D, and / or KRAS-G12V mutant) by contacting the protein in solution with an effective amount of a compound of Formula (I). In some embodiments, the present disclosure provides methods for inhibiting the activity of a mutant KRAS, HRAS, or NRAS protein by contacting a cell, tissue, or organ expressing the protein of interest. In some embodiments, the present disclosure provides methods of inhibiting the activity of a protein in a subject, including but not limited to, rodents and mammals (e.g., humans), by administering to the subject an effective amount of a compound of Formula (I).

[0083] Combination therapy One or more additional pharmacologically active agents may be administered in combination with a compound of formula (I) (or a pharmaceutically acceptable salt thereof). The term "additional active agent(s)" is intended to mean a pharmaceutically active agent(s) that is / are different from the compound of formula (I) and includes prodrugs that are converted into a pharmaceutically active form after administration. The term "additional active agent(s)" also includes free acids, free bases, and pharmaceutically acceptable salts of the additional active agents. Generally, any suitable one or more additional active agents, such as chemotherapeutic agents or therapeutic antibodies, may be used in any combination with a compound of formula (I) in a single dosage formulation (e.g., a fixed-dose combination) or in one or more separate dosage formulations that allow for concurrent or sequential administration of the active agents (co-administration of separate active agents) to a subject. The compound of formula (I) (or a pharmaceutically acceptable salt thereof) may also be administered in combination with radiation therapy, hormone therapy, surgery, or immunotherapy.

[0084] The present application also provides methods for combination therapy, where the additional active agent is known to modulate other pathways, or other components of the same pathway, or even overlapping sets of target enzymes used in combination with a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In one embodiment, such therapy includes, but is not limited to, the combination of one or more compounds of Formula (I) with chemotherapeutic agents, immunotherapeutic agents, hormonal and antihormonal agents, targeted therapeutic agents, and antiangiogenic agents to provide a synergistic or additive therapeutic effect. In another embodiment, such therapy includes radiation treatment to provide a synergistic or additive therapeutic effect.

[0085] Examples of additional active agents (i.e., additional anti-cancer agents) include chemotherapeutic agents (e.g., cytotoxic agents), immunotherapeutic agents, hormonal and anti-hormonal agents, targeted therapy agents, and anti-angiogenic agents. Many anti-cancer agents can be classified into one or more of these groups. Although some specific anti-cancer agents are categorized herein into specific groups or subgroups, many such agents may also be included in one or more other groups or subgroups, as would be understood in the state of the art. It should be understood that the categorization of specific agents herein into specific groups is not intended to be limiting. Many anti-cancer agents are currently known in the art and can be used in combination with the compounds of the present disclosure.

[0086] Furthermore, an agent may be an agonist, antagonist, allosteric modulator, toxin, or more generally, may act to inhibit or stimulate its target (e.g., activation or inhibition of a receptor or enzyme). For example, suitable for use are one or more agents (e.g., antibodies, antigen-binding regions, or soluble receptors) that specifically bind to and inhibit the activity of a growth factor, such as antagonists of hepatocyte growth factor (HGF, also known as scatter factor), and antibodies or antigen-binding regions that specifically bind to its receptor "c-met."

[0087] In one embodiment, the additional anti-cancer agent is a chemotherapeutic agent, an immunotherapeutic agent, a hormone, an antihormonal agent, a targeted therapy, or an anti-angiogenic agent (or angiogenesis inhibitor). In one embodiment, the additional anti-cancer agent is a chemotherapeutic agent, an immunotherapeutic agent, a hormone, an anti-hormonal agent, a plant alkaloid, an alkylating agent, an antimetabolite, a platinum-based analog, an enzyme, a topoisomerase inhibitor, a retinoid, an aziridine, an antibiotic, a hormone, an anti-hormonal agent, an anti-estrogen, an anti-androgen, an adrenocortical suppressant, an androgenic agent, a targeted therapy, an immunotherapeutic agent, a biological response modifier, a cytokine inhibitor, a tumor vaccine, a monoclonal antibody, an immune checkpoint inhibitor, an anti-PD-1 agent, an anti-PD-L1 agent, a colony-stimulating factor, an immunomodulatory agent, an immunomodulatory imid drug (IMiD), an anti-CTLA4 agent, an anti-LAG1 agent, an anti-OX40 agent, a GITR agonist, a CAR-T cell, a BiTE, a signal transduction inhibitor, an enhancer, an anti-inflammatory drug ... growth factor inhibitors, tyrosine kinase inhibitors, EGFR inhibitors, histone deacetylase (HDAC) inhibitors, proteasome inhibitors, cell cycle inhibitors, anti-angiogenic agents, matrix metalloproteinase (MMP) inhibitors, hepatocyte growth factor inhibitors, TOR inhibitors, KDR inhibitors, VEGF inhibitors, HIF-1α inhibitors, HIF-2α inhibitors, fibroblast growth factor (FGF) inhibitors, RAF inhibitors, MEK inhibitors, ERK inhibitors, PI3K inhibitors, AKT inhibitors, MCL-1 inhibitors, BCL-2 inhibitors, SHP2 inhibitors, HER-2 inhibitors, BRAF-inhibitors, gene expression modulators, autophagy inhibitors, apoptosis inducers, antiproliferative agents and glycolysis inhibitors.

[0088] In one embodiment, the additional anti-cancer agent is a chemotherapeutic agent, non-limiting examples of which include antimitotic agents and plant alkaloids, alkylating agents, antimetabolites, platinum-based analogs, enzymes, topoisomerase inhibitors, retinoids, aziridines, and antibiotics.

[0089] Non-limiting examples of antimitotic drugs and plant alkaloids include taxanes such as cabazitaxel, docetaxel, larotaxel, ortataxel, paclitaxel, and tesetaxel; demecolcine; epothilones; eribulin; etoposide (VP-16); etoposide phosphate; navelbine; noscapine; teniposide; thaliblastine; vinblastine; vincristine; vindesine; vinflunine; and vinorelbine.

[0090] Non-limiting examples of alkylating agents include nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, cytophosphane, estramustine, ifosfamide, mannomustine, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, tris(2-chloroethyl)amine, trofosfamide, and uracil mustard; alkylsulfonic acids such as busulfan, improsulfan, and piposulfan; nitrosoureas such as carmustine, chlorozotocin, These include fotemustine, lomustine, nimustine, ranimustine, streptozotocin, and TA-07; ethyleneimines and methylamelamines such as altretamine, thiotepa, triethylenemelamine, triethylenethiophosphaoramide, triethylenephosphoramide, and trimethylolomelamine; ambamustine; bendamustine; dacarbazine; etoglucide; irofulven; mafosfamide; mitobronitol; mitolactol; pipobroman; procarbazine; temozolomide; treosulfan; and triazicon.

[0091] Non-limiting examples of antimetabolites include folic acid analogs such as aminopterin, denopterin, edatrexate, methotrexate, pteropterin, raltitrexed, and trimetrexate; purine analogs such as 6-mercaptopurine, 6-thioguanine, fludarabine, forodesine, thiamiprine, and thioguanine; pyrimidine analogs such as 5-fluorouracil (5-FU), 6-azauridine, ancitabine, azacitidine, capecitabine, carmofu, 3-aminopyridine-2-carboxaldehyde thiosemicarbazone; broxuridine; cladribine; cyclophosphamide; cytarabine; emitefluridine; hydroxyurea; mercaptopurine; nelarabine; pemetrexed; pentostatin; tegafur; and troxacitabine.

[0092] Non-limiting examples of platinum-based analogs include carboplatin, cisplatin, cycloplatin, heptaplatin, lobaplatin, nedaplatin, oxaliplatin, satraplatin, and triplatin tetranitrate.

[0093] Non-limiting examples of enzymes include asparaginase and pegaspargase.

[0094] Non-limiting examples of topoisomerase inhibitors include acridine carboxamide, amonafide, amsacrine, belotecan, elliptinium acetate, exatecan, indolocarbazole, irinotecan, lurtotecan, mitoxantrone, razoxane, rubitecan, SN-38, sobuzoxane, and topotecan.

[0095] Non-limiting examples of retinoids include alitretinoin, bexarotene, fenretinide, isotretinoin, liarozole, R11 retinamide, and tretinoin.

[0096] Non-limiting examples of aziridines include benzodopa, carboquone, meturedopa, and uredopa.

[0097] Non-limiting examples of antibiotics include intercalating antibiotics; anthracenediones; anthracycline antibiotics such as aclarubicin, amrubicin, daunomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, menogaril, nogalamycin, pirarubicin, and valrubicin; 6-diazo-5-oxo-L-norleucine; aclacinomycin; actinomycin; autramycin; azaserine; bleomycin; cactinomycin; calicheamicin; carabicin; carminomycin; These include carzinophilin; chromomycin; dactinomycin; detorubicin; esorubicin; esperamicin; geldanamycin; marcellomycin; mitomycin; mitomycin C; mycophenolic acid; olivomycin; novantrone; peplomycin; porfiromycin; potfiromycin; puromycin; chelamycin; rebeccamycin; rodorubicin; streptonigrin; streptozocin; tanespimycin; tubercidin; ubenimex; zinostatin; zinostatin stimalamer; and zorubicin.

[0098] In one embodiment, the additional anti-cancer agent is a hormonal agent and / or an anti-hormonal agent (i.e., a hormone therapy drug). Non-limiting examples of hormonal agents and anti-hormonal agents include anti-androgens, such as abiraterone, apalutamide, bicalutamide, darolutamide, enzalutamide, flutamide, goserelin, leuprolide, and nilutamide; anti-estrogens, such as 4-hydroxytamoxifen, aromatase inhibitor 4(5)-imidazole, EM-800, fosfestrol, fulvestrant, keoxifene, LY 117018, onapristone, raloxifene, tamoxifen, toremifene, and trioxifene; adrenocortical suppressants such as aminoglutethimide, dexaminoglutethimide, mitotane, and trilostane; androgens such as calsterone, dromostanolone propionate, epitiostanol, mepitiostane, and testolactone; abarelix; anastrozole; cetrorelix; deslorelin; exemestane; fadrozole; finasteride; formestane; histrelin (RL 0903); human chorionic gonadotropin; lanreotide; LDI 200 (Milkhaus); letrozole; leuprorelin; mifepristone; nafarelin; nafoxidine; osaterone; prednisone; thyrotropin alfa; and triptorelin.

[0099] In one embodiment, the additional anti-cancer agent is an immunotherapeutic agent (i.e., an immunotherapeutic drug). Non-limiting examples of immunotherapeutic agents include biological response modifiers, cytokine inhibitors, tumor vaccines, monoclonal antibodies, immune checkpoint inhibitors, colony-stimulating factors, and immunomodulatory agents.

[0100] Non-limiting examples of biological response modifiers, e.g., cytokine inhibitors (cytokines), such as interferons and interleukins, include interferon alpha / interferon alpha, such as interferon alpha-2, interferon alpha-2a, interferon alpha-2b, interferon alpha-nl, interferon alpha-n3, interferon alfacon-1, pegylated interferon alpha-2a, pegylated interferon alpha-2b, and leukocyte interferon alpha; interferon beta, such as interferon beta-1a and interferon beta-1b; interferon gamma, such as natural interferon gamma-1a and interferon gamma-1b; aldesleukin; interleukin-1 beta; interleukin-2; oprelvekin; sonermin; tasonermin; and virulizin.

[0101] Non-limiting examples of tumor vaccines include APC 8015, AVICINE, bladder cancer vaccine, cancer vaccine (Biomira), gastrin 17 immunogen, Maruyama vaccine, melanoma lysate vaccine, melanoma tumor lysate vaccine (New York Medical College), melanoma vaccine (New York University), melanoma vaccine (Sloan Kettering Institute), TICE® BCG (Bacillus Calmette-Guerin), and viral-based melanoma cell lysate vaccine (Royal Newcastle Hospital).

[0102] Non-limiting examples of monoclonal antibodies include abagovomab, adecatumumab, aflibercept, alemtuzumab, blinatumomab, brentuximab vedotin, CA 125 MAb (Biomira), cancer MAb (Japan Pharmaceutical Development), daclizumab, daratumumab, denosumab, edrecolomab, gemtuzumab zogamicin, HER-2 and Fc MAb (Medarex), ibritumomab tiuxetan, idiotype 105AD7 MAb (CRC Technology), idiotype CEA MAb (Trilex), ipilimumab, lintuzumab, LYM-1-iodine-131 MAb (Techni clone), mitumomab, moxetumomab, ofatumumab, polymorphic epithelial mucin-yttrium 90 Examples include MAb (Antisoma), ranibizumab, rituximab and trastuzumab.

[0103] Non-limiting examples of immune checkpoint inhibitors include anti-PD-1 agents or antibodies, such as cemiplimab, nivolumab, and pembrolizumab; anti-PD-L1 agents or antibodies, such as atezolizumab, avelumab, and durvalumab; anti-CTLA-4 agents or antibodies, such as ipilimumab; anti-LAG1 agents; and anti-OX40 agents.

[0104] Non-limiting examples of colony stimulating factors include darbepoetin alfa, epoetin alfa, epoetin beta, filgrastim, granulocyte macrophage colony stimulating factor, lenograstim, relidistim, millimostim, molgramostim, nartograstim, pegfilgrastim, and sargramostim.

[0105] Non-limiting examples of additional immunotherapeutic agents include BiTEs, CAR-T cells, GITR agonists, imiquimod, immunomodulatory imid drugs (IMiDs), mismatched double-stranded RNA (Ampligen), resiquimod, SRL 172, and thymalfasin.

[0106] In one embodiment, the additional anticancer agent is a targeted therapy (i.e., targeted therapy). Targeted therapy agents include, for example, monoclonal antibodies and small molecule drugs. Non-limiting examples of targeted therapy agents include signal transduction inhibitors, growth factor inhibitors, tyrosine kinase inhibitors, EGFR inhibitors, histone deacetylase (HDAC) inhibitors, proteasome inhibitors, cell cycle inhibitors, angiogenesis inhibitors, matrix metalloproteinase (MMP) inhibitors, hepatocyte growth factor inhibitors, TOR inhibitors, KDR inhibitors, VEGF inhibitors, fibroblast growth factor (FGF) inhibitors, MEK inhibitors, ERK inhibitors, PI3K inhibitors, AKT inhibitors, MCL-1 inhibitors, BCL-2 inhibitors, SHP2 inhibitors, HER-2 inhibitors, BRAF inhibitors, gene expression regulators, autophagy inhibitors, apoptosis inducers, antiproliferative agents, and glycolysis inhibitors.

[0107] Non-limiting examples of signal transduction inhibitors include tyrosine kinase inhibitors, multikinase inhibitors, anlotinib, avapritinib, axitinib, dasatinib, dovitinib, imatinib, lenvatinib, lonidamine, nilotinib, nintedanib, pazopanib, pegvisomant, ponatinib, vandetanib, and EGFR inhibitors.

[0108] Non-limiting examples of EGFR inhibitors include small molecule antagonists of EGFR, such as afatinib, brigatinib, erlotinib, gefitinib, lapatinib, and osimertinib; and antibody-based EGFR inhibitors, such as any anti-EGFR antibody or antibody fragment that can partially or completely block activation of EGFR by its natural ligand. Antibody-based EGFR inhibitors include, for example, those described in Modjtahedi, H., et al., 1993, Br. J. Cancer 67:247-253; Teramoto, T., et al., 1996, Cancer 77:639-645; Goldstein et al., 1995, Clin. Cancer Res. 1:1311-1318; Huang, S. M., et al., 1999, Cancer Res. 15:59(8):1935-40; and Yang, X., et al., 1999, Cancer Res. 59:1236-1243; monoclonal antibody Mab E7.6.3 (Yang, 1999, supra); Mab C225 (ATCC deposit number HB-8508) or an antibody or antibody fragment having the binding properties; specific antisense nucleotides or siRNA; afatinib, cetuximab; matuzumab; necitumumab; nimotuzumab; panitumumab; and zalutumumab.

[0109] Non-limiting examples of histone deacetylase (HDAC) inhibitors include belinostat, panobinostat, romidepsin, and vorinostat.

[0110] Non-limiting examples of proteasome inhibitors include bortezomib, carfilzomib, ixazomib, marizomib (salinosporamide a), and oprozomib.

[0111] Non-limiting examples of cell cycle inhibitors, for example CDK inhibitors, include abemaciclib, alvocidib, palbociclib, and ribociclib.

[0112] In one embodiment, the additional anti-cancer agent is an anti-angiogenic agent (or angiogenesis inhibitor), such as, but not limited to, a matrix-metalloproteinase (MMP) inhibitor; a VEGF inhibitor; an EGFR inhibitor; a TOR inhibitor, such as everolimus and temsirolimus; a PDGFR kinase inhibitor, such as crenolanib; a HIF-1α inhibitor, such as PX 478; a HIF-2α inhibitor, such as velzutifan and the HIF-2α inhibitors described in WO 2015 / 035223; a fibroblast growth factor (FGF) or FGFR inhibitor, such as B-FGF and RG 13577; hepatocyte growth factor inhibitors; KDR inhibitors; anti-Ang1 and anti-Ang2 agents; anti-Tie2 kinase inhibitors; Tek antagonists (U.S. Patent Application Publication No. 2003 / 0162712; U.S. Patent No. 6,413,932); anti-TWEAK agents (U.S. Patent No. 6,727,225); ADAM disintegrin domains that antagonize the binding of integrins to their ligands (U.S. Patent Application Publication No. 2003 / 0162712; U.S. Patent No. 6,413,932); 002 / 0042368); anti-eph receptor and / or anti-ephrin antibodies or antigen binding regions (U.S. Patent Nos. 5,981,245; 5,728,813; 5,969,110; 6,596,852; 6,232,447; and 6,057,124); and anti-PDGF-BB antagonists and antibodies or antigen binding regions that specifically bind to PDGF-BB ligands.

[0113] Non-limiting examples of matrix metalloproteinase (MMP) inhibitors include MMP-2 (matrix metalloproteinase 2) inhibitors, MMP-9 (matrix metalloproteinase 9) inhibitors, prinomastat, RO 32-3555, and RS 13-0830. Examples of useful matrix metalloproteinase inhibitors are described, for example, in WO 96 / 33172, WO 96 / 27583, EP 1004578, WO 98 / 07697, WO 98 / 03516, WO 98 / 34918, WO 98 / 34915, WO 98 / 33768, WO 98 / 30566, EP 0606046, and EP 0931788. , WO 90 / 05719, WO 99 / 52910, WO 99 / 52889, WO 99 / 29667, WO 1999 / 007675, EP 1786785, EP 1181017, U.S. Patent No. 2009 / 0012085, U.S. Patent No. 5,863,949, U.S. Patent No. 5,861,510, and EP 0780386. Preferred MMP-2 and MMP-9 inhibitors are those that have little or no activity inhibiting MMP-1. More preferred are those that selectively inhibit MMP-2 and / or MMP-9 over other matrix metalloproteinases (i.e., MAP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13).

[0114] Non-limiting examples of VEGF inhibitors and VEGFR inhibitors include bevacizumab, cediranib, CEP 7055, CP 547632, KRN 633, orantinib, pazopanib, pegaptanib, pegaptanib octasodium, semaxanib, sorafenib, sunitinib, VEGF antagonist (Borean, Denmark), and VEGF-TRAP™.

[0115] Alternatively, the additional anti-cancer agent may be another anti-angiogenic agent, for example, but not limited to, 2-methoxyestradiol, AE 941, alemtuzumab, alpha-D148 Mab (Amgen, US), alphastatin, anecortave acetate, angiocidin, an angiogenesis inhibitor (SUGEN, US), angiostatin, anti-Vn Mab (Crucell, Netherlands), atiprimod, axitinib, AZD 9935, BAY RES 2690 (Bayer, Germany), BC 1 (Genoa Institute of Cancer Research, Italy), beloranib, benefin (Lane Labs, US), cabozantinib, CDP 791 (Celltech Group, UK), chondroitinase AC, cilengitide, combretastatin A4 prodrug, CP 564959 (OSI, US), CV247, CYC 381 (Harvard University, US), E 7820, EHT 0101, endostatin, enzastaurin hydrochloride, ER-68203-00 (IVAX, US), fibrinogen-E fragment, Flk-1 (ImClone Systems, US), form of FLT-1 (VEGFR-1), FR-111142, GCS-100, GW 2286 (GlaxoSmithKline, UK), IL-8, Ilomastat, IM-862, irsogladine, KM-2550 (Kyowa Hakko, Japan), lenalidomide, lenvatinib, MAb alpha5beta3 integrin, second generation (Applied Molecular Evolution USA and MedImmune, US), MAb VEGF (Xenova, UK), marimastat, maspin (Sosei, Japan), metastatin, motuporamine C, M-PGA, ombrulin, OXI4503, PI 88, platelet factor 4, PPI 2458, ramucirumab, rBPI 21 and BPI-derived antiangiogenic (XOMA, US), regorafenib, SC-236, SD-7784 (Pfizer, US), SDX 103 (University of California, San Diego, US), SG 292 (Telios, US), SU-0879 (Pfizer,US), TAN-1120, TBC-1635, tecevatinib, tetrathiomolybdate, thalidomide, thrombospondin 1 inhibitors, Tie-2 ligand (Regeneron, US), tissue factor pathway inhibitors (EntreMed, US), tumor necrosis factor-alpha inhibitors, tumstatin, TZ 93, urokinase-type plasminogen activator inhibitors, vadimezan, vandetanib, vasostatin, vatalanib, VE-cadherin-2 antagonists, xanthorrhizol, XL 784 (Exelixis, US), ziv-aflibercept, and ZD 6126.

[0116] In some embodiments, the additional anti-cancer agent is an additional active agent that disrupts or inhibits the RAS-RAF-ERK or PI3K-AKT-TOR signaling pathways or is an antagonist of PD-1 and / or PD-L1. In some embodiments, the additional anti-cancer agent(s) is a RAF inhibitor, EGFR inhibitor, MEK inhibitor, ERK inhibitor, PI3K inhibitor, AKT inhibitor, TOR inhibitor, MCL-1 inhibitor, BCL-2 inhibitor, SHP2 inhibitor, proteasome inhibitor, or immunotherapeutic agent, such as a monoclonal antibody, immunomodulatory imid drug (IMiD), anti-PD-1, anti-PDL-1, anti-CTLA4, anti-LAG1, and anti-OX40 agent, GITR agonist, CAR-T cell, and BiTE.

[0117] Non-limiting examples of RAF inhibitors include dabrafenib, encorafenib, regorafenib, sorafenib, and vemurafenib.

[0118] Non-limiting examples of MEK inhibitors include binimetinib, CI-1040, cobimetinib, PD318088, PD325901, PD334581, PD98059, refametinib, selumetinib, and trametinib.

[0119] Non-limiting examples of ERK inhibitors include LY3214996, LTT462, MK-8353, SCH772984, lavoxertinib, ulixertinib, and ERKi's such as those described in WO 2017 / 068412.

[0120] Non-limiting examples of PI3K inhibitors include 17-hydroxywortmannin analogs (e.g., WO 06 / 044453); AEZS-136; alpelisib; AS-252424; buparlisib; CAL263; copanlisib; CUDC-907; dactolisib (WO 06 / 122806); demethoxyviridine; duvelisib; GNE-477; GSK1059615; IC87114; idelalisib; INK1117; LY294002; Palomid 529; paxalisib; perifosine; PI-103; PI-103 hydrochloride; pictilisib (e.g., WO 09 / 036,082; WO 09 / 055,730); PIK 90; PWT33597; SF1126; sonolisib; TGI 00-115; TGX-221; XL147; XL-765; wortmannin; and ZSTK474.

[0121] Non-limiting examples of AKT inhibitors include Akt-1-1 (inhibits Aktl) (Barnett et al. (2005) Biochem. J., 385(Pt. 2), 399-408); Akt-1-1,2 (Barnett et al. (2005) Biochem. J. 385(Pt. 2), 399-408); API-59CJ-Ome (e.g., Jin et al. (2004) Br. J. Cancer 91, 1808-12); 1-H-imidazo[4,5-c]pyridinyl compounds (e.g., WO 05011700); indole-3-carbinol and its derivatives (e.g., U.S. Pat. No. 6,656,963; Sarkar and Li (2004) J Nutr. 134(12) Supplement), 3493S-3498S); perifosine, Dasmahapatra et al. (2004) Clin. Cancer Res. 10(15), 5242-52, 2004); phosphatidylinositol ether lipid analogues (e.g., Gills and Dennis (2004) Expert. Opin. Investig. Drugs 13, 787-97); triciribine (Yang et al. (2004) Cancer Res. 64, 4394-9); imidazooxazone compounds such as trans-3-amino-1-methyl-3-[4-(3-phenyl-5H-imidazo[1,2-c]pyrido[3,4-e][1,3]oxazin-2-yl)phenyl]-cyclobutanol hydrochloride (WO 2012 / 137870); afuresertib; capivasertib; MK2206; patasertib, and those disclosed in WO 2011 / 082270 and WO 2012 / 177844.

[0122] Non-limiting examples of TOR inhibitors include deforolimus; ATP-competitive TORC1 / TORC2 inhibitors, such as PI-103, PP242, PP30, and Torin1; TOR inhibitors at the FKBP12 enhancer, rapamycin and its derivatives, such as temsirolimus, everolimus, WO 9409010; and those described in WO 98 / 02441 and WO 01 / 14387. rapalogs such as those disclosed in US Pat. No. 6,235,739, such as AP23573, AP23464, or AP23841; 40-(2-hydroxyethyl)rapamycin, 40-[3-hydroxy(hydroxymethyl)methylpropanoate]-rapamycin; 40-epi-(tetrazolyl)-rapamycin (also known as ABT578); 32-deoxorapamycin; 16-pentynyloxy-32(S)-dihydrorapamycin; rapanycin, as well as other derivatives disclosed in WO 05 / 005434; U.S. Pat. No. 5,258,389, WO 94 / 090101, WO 92 / 05179, U.S. Pat. No. 5,118,677, U.S. Pat. No. 5,118,678, U.S. Pat. No. 5,100,883, U.S. Pat. No. 5,151,413, U.S. Pat. No. 5,120,842, WO 93 / 11 1130, WO 94 / 02136, WO 94 / 02485, WO 95 / 14023, WO 94 / 02136, WO 95 / 16691, WO 96 / 41807, derivatives disclosed in WO 96 / 41807 and U.S. Pat. No. 5,256,790; and phosphorus-containing rapamycin derivatives (e.g., WO 05 / 016252).

[0123] Non-limiting examples of MCL-1 inhibitors include AMG-176, MIK665, and S63845.

[0124] Non-limiting examples of SHP2 inhibitors include those described in WO 2019 / 167000 and WO 2020 / 022323.

[0125] Further non-limiting examples of anti-cancer agents suitable for use include 2-ethylhydrazide, 2,2',2"-trichlorotriethylamine, ABVD, aceglatone, acemannan, aldophosphamide glycoside, alpharazine, amifostine, aminolevulinic acid, anagrelide, ANCER, ancestim, anti-CD22 immunotoxin, anti-tumorigenic herbs, apaziquone, aruglavin, arsenic trioxide, azathioprine, BAM 002 (Novelos), bcl-2 (Genta), bestrabucil, biricodar, bisantrene, bromocriptine, brostallicin, bryostatin, buthionine sulfoximine, calyculin, cell cycle non-specific antineoplastic agents, celmoleukin, clodronate, clotrimazole, cytarabine ocfosfate, DA 3030 (Dong-A), defofamine, denileukin diftitox, dexrazoxane, diazicon, dichloroacetic acid, dilazep, discodermolide, docosanol, doxercalciferol, edelfosine, eflornithine, EL532 (Elan), elfomithine, elsamitrucin, eniluracil, etanidazole, exisulind, ferruginol, folic acid supplements, e.g., florin (f rolinic acid, gacytosine, gallium nitrate, gimeracil / oteracil / tegafur combination (S-1), glycopine, histamine dihydrochloride, HIT diclofenac, HLA-B7 gene therapy drug (Vical), human fetal alpha-fetoprotein, ibandronate, ibandronic acid, ICE chemotherapy regimen, imexon, iobenguane, IT-101 (CRLX101), laniquidar, LC 9018 (Yakult), leflunomide, lentinan, levamisole + fluorouracil, lovastatin, lucantone, masoprocol, melarsoprol, metoclopramide, miltefosine, miproxifene, mitoguazone, mitozolomide, mopidamol, motexafine gadolinium, MX6 (Galderma), naloxone + pentazocine, nitracrine, nolatrexed, NSC 631570Octreotide (Ukrain), olaparib, P-30 protein, PAC-1, palifermin, pamidronate, pamidronic acid, pentosan polysulfate sodium, phenamet, picibanil, pixantrone, platinum, podophyllic acid, porfimer sodium, PSK (polysaccharide K), rabbit antithymocyte polyclonal antibody, rasburiembodiment, retinoic acid, rhenium Re 186 etidronate, romurtide, samarium (153 Sm) lexidronam, sizofiran, sodium phenylacetate, sparfosic acid, spirogermanium, strontium-89 chloride, suramin, swainsonine, talaporfin, tariquidar, tazarotene, tegafur-uracil, temoporfin, tenuazonic acid, tetrachlorodecaoxide, thrombopoietin, ethyl etiopurinse, tirapazamine, TLC ELL-12, tositumomab-iodine 131, trifluridine and tipiracil combination, troponin I (Harvard University, US), urethane, valspodar, verteporfin, zoledronic acid, and zosuquidar.

[0126] The present disclosure further provides a method for using the compound of formula (I) or pharmaceutical composition provided herein in combination with radiation therapy to treat cancer. Techniques for administering radiation therapy are known in the art, and such techniques can be used in the combination therapy described herein. The administration of the compound of formula (I) in this combination therapy can be determined as described herein.

[0127] Radiation therapy can be administered by one of several methods or a combination of methods, including, but not limited to, external beam radiation therapy, internal radiation therapy, interstitial irradiation, stereotactic radiosurgery, systemic radiation therapy, radiotherapy, and permanent or temporary interstitial brachytherapy. The term "brachytherapy," as used herein, refers to radiation therapy delivered by spatially confined radioactive material inserted at or near the diseased site of a tumor or other proliferative tissue within the body. This term is intended to encompass, but is not limited to, exposure to radioactive isotopes (e.g., At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, and radioactive isotopes of Lu). Suitable radiation sources for use as cell conditioners of the present disclosure include both solid and liquid forms. By way of non-limiting example, the radiation source can be a radionuclide, such as I-125, I-131, Yb-169, Ir-192 as a solid source, I-125 as a solid source, or other radionuclide that emits photons, beta particles, gamma radiation, or other therapeutic rays. The radioactive material can also be a fluid made from any solution of radionuclide(s), such as a solution of I-125 or I-131, or a radioactive fluid can be made using a slurry of a suitable fluid containing small particles of a solid radionuclide, such as Au-198 or Y-90. Furthermore, the radionuclide(s) can be made into a gel or radioactive microspheres.

[0128] The present disclosure further provides methods for combination therapy where the additional active agent is known to modulate other pathways, or other components of the same pathway, or even overlapping sets of target enzymes used in combination with a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In one embodiment, such therapy includes, but is not limited to, the combination of one or more compounds of Formula (I) with chemotherapeutic agents, immunotherapeutic agents, hormonal therapy agents, therapeutic antibodies, targeted therapy agents, and radiation treatment to produce synergistic or additive therapeutic effects.

[0129] The compounds of the present disclosure can be used in combination with the drugs disclosed herein or other suitable drugs, depending on the condition being treated. Thus, in some embodiments, one or more compounds of the present disclosure are co-administered with other drugs as described above. When used in combination therapy, the compounds described herein are administered simultaneously with the second drug or separately. This administration in combination can include simultaneous administration of the two drugs in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, the compound of Formula (I) and any of the drugs described above can be formulated together in the same dosage form and administered simultaneously. Alternatively, the compound of Formula (I) and any of the drugs described above can be administered simultaneously, but both drugs are present in separate formulations. Alternatively, the compound of Formula (I) can be administered immediately after the drug described above, or vice versa. In some embodiments of the separate administration protocol, the compound of Formula (I) and any of the drugs described above are administered minutes, hours, or days apart.

[0130] In one aspect of the present disclosure, since treatment of a disease / condition with a combination of pharmaceutically active compounds that can be administered separately is contemplated, the present disclosure further relates to combining separate pharmaceutical compositions into a kit form. The kit comprises two separate pharmaceutical compositions: a compound of Formula (I) and a second pharmaceutical compound. The kit comprises containers for housing the separate compositions, such as divided bottles or divided foil packets. Further examples of containers include syringes, boxes, and bags. In some embodiments, the kit comprises instructions for use of the separate components. The kit form is particularly advantageous when the separate components are preferably administered in different dosage forms (e.g., oral and parenteral), administered at different dosing intervals, or when the prescribing medical professional desires to titrate the individual components of the combination.

[0131] The present disclosure also provides a compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in therapy, or the use of a compound of Formula (I) or a pharmaceutically acceptable salt thereof in therapy. The present disclosure also provides a compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, or the use of a compound of Formula (I) or a pharmaceutically acceptable salt thereof for treating cancer. The present disclosure also provides a compound of Formula (I) or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of cancer, or the use of a compound of Formula (I) or a pharmaceutically acceptable salt thereof for the preparation of a medicament for the treatment of cancer. The present disclosure also provides a compound of Formula (I) or a pharmaceutically acceptable salt thereof and an additional anticancer agent for use in the treatment of cancer, or the use of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and the additional anticancer agent for treating cancer. The present disclosure also provides use of a compound of formula (I) or a pharmaceutically acceptable salt thereof and an additional anticancer agent for the preparation of a medicament for the treatment of cancer, or use of a compound of formula (I) or a pharmaceutically acceptable salt thereof and an additional anticancer agent for the preparation of a medicament for the treatment of cancer. The present disclosure also provides a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer, or use of a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof for treating cancer. The present disclosure also provides a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and an additional anti-cancer agent for use in treating cancer, or use of a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and the additional anti-cancer agent for treating cancer.

[0132] Methods for preparing compounds of the present disclosure The compounds described herein can be prepared according to the procedures of the following schemes and examples using appropriate materials, and are further exemplified by the following specific examples. The examples further describe the details of the preparation of the compounds of the present disclosure. Those skilled in the art will readily understand that known variations of the conditions and processes of the following preparative procedures can be used to prepare these compounds. For example, in some cases, the order of carrying out the steps of the reaction schemes can be changed to facilitate the reaction or to avoid unwanted reaction products. These examples are provided for the purpose of further illustration only and are not intended to limit the disclosure.

[0133] Throughout the synthetic schemes and examples, the following abbreviations and acronyms may be used unless otherwise specified: s: singlet, d: doublet, t: triplet, q: quartet, sep: septet, dd: double doublet, dt: double triplet, td: triple doublet, tt: triple triplet, ddd: double double doublet, ddt: double double triplet, dtd: double triple doublet, tdd: triple double doublet, m: multiplet, br: broad, brs: broad singlet, tert: tertiary, DMSO-d: deuterated dimethyl sulfoxide, CDCl: deuterated chloroform, CDOD: deuterated methanol, THF: tetrahydrofuran, DMF: N,N-dimethylformamide, DMSO: dimethyl sulfoxide, DCM: dichloromethane, IPE: diisopropyl ether, MTBE : methyl tert-butyl ether, EtOAc: ethyl acetate, AcOH: acetic acid, TFA: trifluoroacetic acid, MeOH: methanol, EtOH: ethanol, DIAD: diisopropyl azodicarboxylate, TMAD: N,N,N',N'-tetramethylazodicarboxamide, EtN: triethylamine, DIEA: N,N-diisopropylethylamine, RT: room temperature, NIS: N-iodosuccinimide, Pd(dppf)Cl2: [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II), and HATU: 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, KOAc: potassium acetate.

[0134] Reagents used in the examples are commercially available unless otherwise noted. Prepacked columns manufactured by Shoko Scientific Co., Ltd. or Biotage were used for silica gel column chromatography and basic silica gel column chromatography. NMR spectra were obtained using an AVANCE NEO 400 spectrometer (400 MHz, BRUKER) and an AVANCE III HD 500 spectrometer (500 MHz, BRUKER). For deuterated solvents containing tetramethylsilane, tetramethylsilane was used as the internal standard. In other cases, measurements were performed using the NMR solvent as the internal standard. All δ values ​​are in ppm. Microwave reactions were performed using an Initiator™ manufactured by Biotage. A Waters XSelect CSH C18 OBD Prep column was used for preparative reversed-phase HPLC.

[0135] Benzyl 2-methyl-5-oxoazepane-1-carboxylate (Int-J003-P1) [ka]

[0136] Step A: 1,4-Dioxaspiro[4.5]decan-8-one O-ethylsulfonyloxime (Int-J001) To a solution of 1,4-dioxaspiro[4.5]decan-8-one oxime (3.00 g, 17.5 mmol) and trimethylamine (2.96 mL, 21.0 mmol) in dichloromethane (30 mL) was added ethanesulfonyl chloride (1.84 mL, 19.3 mmol) at -15 °C, and the mixture was stirred at the same temperature for 0.5 h. Water (15 mL) was added to the reaction mixture, and the organic phase was separated. The organic phase was washed with hydrochloric acid (1 M, 15 mL), saturated sodium bicarbonate solution (15 mL), and brine (15 mL), and the organic phase was dried over sodium sulfate. The product solution was filtered, and the filtrate was used in the next step without further purification. ESI-MS m / z [M+H] + 264.

[0137] Step B: Benzyl 9-methyl-1,4-dioxa-8-azaspiro[4.6]undecane-8-carboxylate (Int-J002) The above solution of 1,4-dioxaspiro[4.5]decan-8-one O-ethylsulfonyloxime (Int-J001) in dichloromethane was added dropwise to a stirred solution of trimethylaluminum (1.40 M in hexane, 25.0 mL, 35.0 mmol) at −78°C. The reaction mixture was warmed to 0°C and stirred for 1 hour. Diisobutylaluminum hydride (1.00 M in hexane, 26.3 mL, 26.3 mmol) was added dropwise to the mixture, and the mixture was stirred at 0°C for 1 hour. The reaction mixture was diluted with dichloromethane (15 mL), and sodium fluoride (10.5 g, 0.25 mol) and water (4.4 mL) were added to the reaction mixture. The mixture was stirred vigorously at room temperature for 1 hour and filtered. The filtrate was concentrated under reduced pressure, and ethyl acetate (50 mL) and water (50 mL) were added to the residue. Sodium bicarbonate (30 g, 0.36 mol) and benzyl chloroformate (11.9 g, 70.0 mmol) were added to the mixture at room temperature, and the reaction mixture was stirred for 1 hour. The organic phase was separated, washed with water, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 40%, ethyl acetate gradient / hexane) to give benzyl 9-methyl-1,4-dioxa-8-azaspiro[4.6]undecane-8-carboxylate (Int-J002) (3.60 g). ESI-MS m / z [M+H] + 306.

[0138] Step C: Benzyl 2-methyl-5-oxoazepane-1-carboxylate (Int-J003) Benzyl 9-methyl-1,4-dioxa-8-azaspiro[4.6]undecane-8-carboxylate ( Int-J002 To a solution of Int-J003 (21.0 g, 68.8 mmol) in acetone (300 mL) and water (100 mL) was added toluene-4-sulfonic acid hydrate (19.6 g, 103 mmol). After stirring the mixture at 40 °C for 7 h, the reaction mixture was extracted with EtOAc, washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 50%, EtOAc gradient / hexanes) to give benzyl 2-methyl-5-oxoazepane-1-carboxylate (Int-J003) (14.7 g). ESI-MS m / z [M+H]+ 262.

[0139] Step D: Benzyl 2-methyl-5-oxoazepane-1-carboxylate (Int-J003-P1 and Int-J003-P2) Racemic benzyl 2-methyl-5-oxoazepane-1-carboxylate ( Int-J003 ) was isolated and chiral benzyl 2-methyl-5-oxoazepane-1-carboxylate ( Int-J003-P1 , under the analytical conditions described below) is used for the synthesis.

[0140] Analytical conditions: column (CHIRALCEL OD-H, 4.6 mm ID x 250 mm L, 5 μL), eluent (hexane / 2-propanol, 700 / 300 (volume ratio)), flow rate (1.0 mL / min), temperature (30°C), concentration (0.5 mg / mL), injection volume (10 μL).

[0141] Benzyl 2-methyl-5-oxoazepane-1-carboxylate ( Int-J003-P1 ) retention time: 6.053 min.

[0142] Benzyl 2-methyl-5-oxoazepane-1-carboxylate ( Int-J003-P2 ) retention time: 7.945 min.

[0143] 5,5-Difluoro-2-methylazepane (Int-K002) [ka]

[0144] To a solution of benzyl 2-methyl-5-oxoazepane-1-carboxylate (Int-J003-P1) (200 mg, 0.765 mmol) in dichloromethane (1.5 mL) was added dimethylaminosulfur trifluoride (0.700 mL, 6.41 mmol) at 0 °C. The mixture was stirred at room temperature for 2 h and then quenched by the addition of saturated aqueous Na2CO3. The reaction mixture was extracted with CHCl3, washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 50%, EtOAc gradient / hexanes) to give the difluorinated compound.

[0145] To a solution of the difluorinated compound in ethanol (10 mL) was added Pd(OH)2 / C (269 mg, 0.383 mmol). The reaction mixture was purged with H2 gas and evacuated three times and stirred at room temperature for 1 h. The reaction mixture was filtered and concentrated under reduced pressure to give 5,5-difluoro-2-methylazepane (Int-K002) (20.0 mg). ESI-MS m / z [M+H] + 150.

[0146] 7-Methylazepan-4-ol hydrochloride (Int-K003-A) [ka]

[0147] Step A: Benzyl 5-hydroxy-2-methylazepane-1-carboxylate (Int-K011-P1 and P2) To a solution of benzyl 2-methyl-5-oxoazepane-1-carboxylate (Int-J003P1) (400 mg, 1.53 mmol) in MeOH (16 mL) was added a 1.0 M solution of lithium tri-sec-butylborohydride in THF (1.53 mL, 1.53 mmol) at 0 °C. The mixture was stirred at room temperature for 1 h and then quenched by the addition of HO. The reaction mixture was extracted with EtOAc, washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 50%, EtOAc gradient / hexanes) to give benzyl 5-hydroxy-2-methylazepane-1-carboxylate (Int-K011-P1, first elution): ESI-MS m / z [M+H] + 264 and benzyl 5-hydroxy-2-methylazepane-1-carboxylate (Int-K011-P2, second elution): ESI-MS m / z [M+H] + I got 264.

[0148] Step B: 7-Methylazepan-4-ol hydrochloride (Int-K003-A)To a solution of benzyl 5-hydroxy-2-methylazepane-1-carboxylate (Int-K011-P1, first elution) (79.0 mg, 0.300 mmol) in ethanol (3.0 mL) was added Pd(OH)2 / C (105 mg, 0.150 mmol). The reaction mixture was purged with H2 gas and evacuated three times and stirred at room temperature for 30 minutes. The reaction mixture was filtered, and a 4.0 M solution of HCl in dioxane (1.0 mL) was added to the filtrate. The filtrate was concentrated under reduced pressure to give 7-methylazepan-4-ol hydrochloride (Int-K003-A). ESI-MS m / z [M+H] + 130.

[0149] 7-Methylazepan-4-ol hydrochloride (Int-K003-B) [ka]

[0150] 7-Methylazepan-4-ol hydrochloride (Int-K003-B) was synthesized via the same route as 7-methylazepan-4-ol hydrochloride (Int-K003-A), using benzyl 5-hydroxy-2-methylazepan-1-carboxylate (Int-K011-P2, second elution) instead of benzyl 5-hydroxy-2-methylazepan-1-carboxylate (Int-K011-P1, first elution). ESI-MS m / z [M+H] + 130.

[0151] 5-Fluoro-2-methylazepane hydrochloride (Int-K004-A) [ka]

[0152] To a solution of benzyl 5-hydroxy-2-methylazepane-1-carboxylate (Int-K011-P1, first elution) (175 mg, 0.665 mmol) in dichloromethane (1.3 mL) was added dimethylaminosulfur trifluoride (0.143 mL, 1.31 mmol) at 0 °C. The mixture was stirred at room temperature for 1 h and then quenched by the addition of saturated aqueous Na2CO3. The reaction mixture was extracted with CHCl3, washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 50%, EtOAc gradient / hexanes) to give the fluorinated compound.

[0153] To a solution of the fluorinated compound in ethanol (5.0 mL) was added Pd(OH)2 / C (69.3 mg, 0.0987 mmol). The reaction mixture was purged with H2 gas and evacuated three times and stirred at room temperature for 1 hour. The reaction mixture was filtered, and a 4.0 M solution of HCl in dioxane (1.0 mL) was added to the filtrate. The filtrate was concentrated under reduced pressure to give 5-fluoro-2-methylazepane hydrochloride (Int-K004-A). ESI-MS m / z [M+H] + 132.

[0154] 5-Fluoro-2-methylazepane hydrochloride (Int-K004-B) [ka]

[0155] 5-Fluoro-2-methylazepane hydrochloride (Int-K004-B) was synthesized via the same route as 5-fluoro-2-methylazepane hydrochloride (Int-K004-A), using benzyl 5-hydroxy-2-methylazepane-1-carboxylate (Int-K011-P2, second elution) instead of benzyl 5-hydroxy-2-methylazepane-1-carboxylate (Int-K011-P1, first elution). ESI-MS m / z [M+H] + 132.

[0156] 2-(3-Hydroxypiperidin-3-yl)acetonitrile hydrochloride (Int-XX023) [ka]

[0157] To a solution of benzyl 1-oxa-5-azaspiro[2.5]octane-5-carboxylate (950 mg, 3.84 mmol) in EtOH (20 mL) and water (20 mL) was added sodium cyanide (282 mg, 5.76 mmol). After stirring the mixture at room temperature for 4 hours, the reaction was extracted with EtOAc, washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 50%, EtOAc gradient / hexanes) to give the cyanide compound.

[0158] To a solution of the cyanide compound (110 mg, 0.401 mmol) in ethanol (2.0 mL) was added Pd(OH)2 / C (141 mg, 0.200 mmol). The reaction mixture was purged with H2 gas and evacuated three times and stirred at room temperature for 1 hour. The reaction mixture was filtered, and a 4.0 M solution of HCl in dioxane (1.0 mL) was added to the filtrate. The filtrate was concentrated under reduced pressure to give 2-(3-hydroxypiperidin-3-yl)acetonitrile hydrochloride (Int-XX023). ESI-MS m / z [M+H] + 141.

[0159] 3-Hydroxy-8-iodo-1-naphthoic acid (Int-T002) [ka]

[0160] Step A: 3-Amino-8-iodo-1-naphthoic acid (Int-T001) To a mixture of 8-iodo-3-nitro-1-naphthoic acid (1 g, 2.9 mmol) in ethyl acetate (40 mL) and ethanol (15 mL) was added 5% Rh / C (0.5 g), and the flask was charged with H. The mixture was stirred at room temperature. After 24 h, the reaction mixture was filtered through a pad of Celite, and the filtrate was concentrated to dryness to give crude 3-amino-8-iodo-1-naphthoic acid (Int-T001) (0.94 g) as a brown solid, which was used in the next step without purification. 1H NMR (400 MHz, DMSO-d6) δ = 5.71 (brs, 2H), 6.87 (d, J=2.4 Hz, 1H), 7.00 (dd, J= 8.1, 7.3 Hz, 1H), 7.12 (d, J=2.4 Hz, 1H), 7.60 (d, J= 8.2 Hz, 1H), 7.8 (d, J= 7.3 Hz, 1H), 13.14 (brs, 1H). LCMS (ESI): m / z [M+H] + 314.

[0161] Step B: 3-Hydroxy-8-iodo-1-naphthoic acid (Int-T002) To an ice-cooled mixture of crude 3-amino-8-iodo-1-naphthoic acid (Int-T001) (0.94 g) in 1 M aqueous sulfuric acid (38 mL) was slowly added dropwise a solution of sodium nitrite (0.228 g, 3.3 mmol) in 1 mL of water. The mixture was stirred for 1 hour and then warmed to room temperature. The reaction mixture was added dropwise to refluxing 40% aqueous sulfuric acid (108 mL). The reaction mixture was heated to reflux for 1 hour and then rapidly filtered hot through a layer of glass wool to remove insoluble char. The filtrate was cooled to room temperature, resulting in the formation of a precipitate. The precipitate was collected by filtration and washed with water to give 3-hydroxy-8-iodo-1-naphthoic acid (Int-T002) (0.54 g). 1 H NMR (400 MHz, DMSO-d6) δ =7.13 (dd, J= 8.1, 7.3 Hz, 1H), 7.22 (d, J=2.8 Hz, 1H), 7.24 (d, J=2.8 Hz, 1H), 7.82 (dd, J= 8.3, 0.8 Hz, 1H), 8.01 (dd, J= 7.3, 1.2 Hz, 1H), 10.22 (brs, 1H). LCMS (ESI): m / z [M+H] + 315.

[0162] (2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-hydroxy-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone dihydrochloride (Int-L004) [ka]

[0163] Step A: tert-Butyl 2-chloro-4-((2-nitrobenzyl)oxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-L001) Cesium carbonate (92.6 g, 0.284 mol) was added to a stirred solution of tert-butyl 2,4-dichloro-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (55.0 g, 0.190 mol) and (2-nitrophenyl)methanol (29.0 g, 0.190 mol) in toluene (550 mL), and the mixture was warmed to 85°C and stirred for 14 hours. The mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. Acetonitrile (400 mL) and water (200 mL) were added to the residue, and the mixture was suspended for 2 days. The solid was filtered and suspended in ethyl acetate (400 mL) at 85°C for 1 hour, then cooled to room temperature. The solid was filtered and dried under vacuum to give tert-butyl 2-chloro-4-((2-nitrobenzyl)oxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate ( Int-L001 ) (21g) was obtained. ESI-MS m / z [M+H] + 407, 409.

[0164] Step B: N,N-Dimethyl-1-(1-(((4-((2-nitrobenzyl)oxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanamine (Int-L002) To a mixture of tert-butyl 2-chloro-4-((2-nitrobenzyl)oxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-L001) (10.0 g, 24.6 mmol), (1-((dimethylamino)methyl)cyclopropyl)methanol (4.13 g, 32.0 mmol), rac-BINAP (2.30 g, 3.69 mmol), and palladium(II) acetate (414 mg, 1.84 mmol) in toluene (100 mL) was added cesium carbonate (20.0 g, 61.5 mmol), and the mixture was warmed to 130 °C and stirred for 3 h. The mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (0% to 100%, ethyl acetate gradient / hexane) to give the coupled product. To a solution of the coupled product in dichloromethane (50 mL) was added trifluoroacetic acid (30 mL), and the mixture was stirred for 1 h. Trifluoroacetic acid was removed under reduced pressure, and the residue was purified by flash NH-silica gel chromatography (0% to 50%, methanol gradient / ethyl acetate) to give N,N-dimethyl-1-(1-(((4-((2-nitrobenzyl)oxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanamine (Int-L002) (7.94 g). ESI-MS m / z [M+H] + 400.

[0165] Step C: (2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-((2-nitrobenzyl)oxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Int-L003) To a mixture of 3-hydroxy-8-iodo-1-naphthoic acid (Int-T002) (5.19 g, 16.5 mmol), 1-hydroxybenzotriazole monohydrate (2.53 g, 16.5 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.26 g, 16.5 mmol), and N,N-diisopropylethylamine (7.02 mL, 41.3 mmol) in N,N-dimethylformamide (55 mL) was added N,N-dimethyl-1-(1-(((4-((2-nitrobenzyl)oxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanamine (Int-L002) (5.50 g, 13.8 mmol), and the mixture was stirred at room temperature for 7 hours. The mixture was diluted with ethyl acetate, washed with water, and concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (0% to 60%, methanol gradient / ethyl acetate) to give (2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-((2-nitrobenzyl)oxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Int-L003) (8.77 g). ESI-MS m / z [M+H] + 696.

[0166] Step D: (2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-hydroxy-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone dihydrochloride (Int-L004) To a stirred suspension of (2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-((2-nitrobenzyl)oxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Int-L003) (7.00 g, 10.1 mmol) and iron (powder, 2.81 g, 50.3 mmol) in tetrahydrofuran (56 mL) was added hydrochloric acid (1 M, 56.0 mL, 5.56 mmol), and the mixture was stirred at room temperature for 2 hours. The mixture was diluted with methanol (100 mL) and ethyl acetate (100 mL), filtered, and concentrated to dryness. To the residue was added ethyl acetate (30 mL) and methanol (3 mL), and the mixture was suspended for 1 hour. The precipitate was filtered and dried under vacuum to give (2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-hydroxy-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone dihydrochloride (Int-L004) (6.80 g). ESI-MS m / z [M+H] + 561.

[0167] (4-Hydroxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone dihydrochloride (Int-L005) [ka]

[0168] (4-Hydroxy-2-((1-(morpholinomethyl)cyclopropyl)methoxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone dihydrochloride (Int-L005) was synthesized via a similar route to (2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-hydroxy-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone dihydrochloride (Int-L004), except that (1-(morpholinomethyl)cyclopropyl)methanol was used instead of (1-(dimethylamino)methyl)cyclopropyl)methanol. ESI-MS m / z [M+H]+ 603

[0169] (R)-(1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methanol (Int-UU8-1) [ka]

[0170] Step A: 2-[(acetyloxy)methyl]prop-2-en-1-yl acetate (Int-UU1) A 5-L four-necked round-bottom flask was charged with 3-chloro-2-(chloromethyl)prop-1-ene (600 g, 4.80 mol), triethylamine (1.46 kg, 14.40 mol), and acetic acid (721 g, 12.0 mol). The resulting solution was stirred at 70 °C overnight. The reaction mixture was cooled to room temperature and quenched by adding 3 L of water. The resulting solution was extracted with ethyl acetate (3 x 1 L), and the combined organic phase was washed with brine solution (2 x 1 L). The organic phase was dried over anhydrous sodium sulfate. The dried solution was filtered, and the filtrate was concentrated. The residue was purified on a silica gel column with ethyl acetate / petroleum ether (1:6) to give 2-[(acetyloxy)methyl]prop-2-en-1-yl acetate (Int-UU1).

[0171] Step B: [1-[(acetyloxy)methyl]-2,2-difluorocyclopropyl]methyl acetate (Int-UU2) A solution of 2-[(acetyloxy)methyl]prop-2-en-1-yl acetate (Int-UU1) (600 g, 3.48 mol) in diglyme (5 L) was placed in a 20 L four-necked round-bottom flask maintained under an inert atmosphere of nitrogen. A solution of ClCF2CO2Na (2.65 kg, 17.4 mol) in diglyme (5 L) was then added dropwise with stirring at 180 °C over 5 hours. The resulting solution was stirred at 180 °C for 1 hour. The reaction mixture was cooled to room temperature and quenched by the addition of H2O (5 L). The resulting solution was extracted with petroleum ether (4 x 2 L), and the organic phases were combined. The combined organic phases were washed with water (3 x 2 L) and dried over anhydrous sodium sulfate. The dried solution was filtered and the filtrate was concentrated to dryness to give [1-[(acetyloxy)methyl]-2,2-difluorocyclopropyl]methyl acetate (Int-UU2), which was used directly in the next step without purification.

[0172] Step C: [2,2-Difluoro-1-(hydroxymethyl)cyclopropyl]methanol (Int-UU3) A 20-liter four-necked round-bottom flask was charged with [1-[(acetyloxy)methyl]-2,2-difluorocyclopropyl]methyl acetate (Int-UU2) (800 g, 3.60 mol), MeOH (10 L), and K2CO3 (995 g, 7.20 mol). The resulting solution was stirred at room temperature overnight. The solids were filtered off. The filtrate was concentrated. The resulting mixture was diluted by adding water (2 L). The resulting solution was extracted with ethyl acetate (5 x 1 L). The organic phases were combined and dried over anhydrous sodium sulfate. The dried solution was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with ethyl acetate / petroleum ether (1:1) to give [2,2-difluoro-1-(hydroxymethyl)cyclopropyl]methanol (Int-UU3). 1 H NMR (300 MHz, DMSO-d6) δ 1.30 (t, J=8.8 Hz, 2H), 3.52 (m, 4H), 4.79 (t, J=5.6 Hz, 2H).

[0173] Step D: (1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methanol (Int-UU4) A 500 mL single-necked round-bottom flask equipped with a nitrogen sparging adapter was purged with nitrogen and then charged with sodium hydride (4.52 g, 113 mmol) and N,N-dimethylformamide (100 mL). The suspension was cooled to 0 °C. Solid [2,2-difluoro-1-(hydroxymethyl)cyclopropyl]methanol (Int-UU3) (12.0 g, 87 mmol) was added in small portions. The mixture was stirred and allowed to warm to room temperature over 1 h. The resulting reaction mixture was cooled to 0 °C and treated with a solution of benzyl bromide (10.3 mL, 87 mmol) in N,N-dimethylformamide (10 mL). The mixture was stirred at room temperature for 1 h and then treated with saturated aqueous ammonium chloride (10 mL) and water (10 mL). The mixture was partitioned between ethyl acetate (75 mL) and water (75 mL). The organic phase was washed with 1 wt% aqueous LiCl (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Purification by column chromatography on silica gel (220 g, 0% to 40% EtOAc / hexane) gave (1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methanol (Int-UU4). 1 H NMR (499 MHz, methanol-d4) δ 7.39 ? 7.32 (m, 4H), 7.32 ? 7.26 (m, 1H), 4.62 ? 4.49 (m, 2H), 3.79 ? 3.64 (m, 3H), 3.60 (dd, J = 10.4, 2.1 Hz, 1H), 1.35 (dddd, J = 29.1, 12.5, 8.0, 4.5 Hz, 2H).

[0174] Step E: (1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methanol (Int-UU5-1) Racemic (1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methanol (Int-UU4) was resolved using SFC chiral chromatography (AD-H (21 mm × 250 mm, 5 μm; conditions: 5% MeOH + 0.1% NH OH and 5% HO)) to give (1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methanol (Int-UU5-1, peak 1). 1 H NMR (499 MHz, methanol-d4) δ 7.39 ? 7.26 (m, 5H), 4.58 ? 4.51 (m, 2H), 3.78 ? 3.67 (m, 3H), 3.60 (dd, J = 10.4, 2.0 Hz, 1H), 1.35 (dddd, J = 28.4, 12.5, 8.0, 4.5 Hz, 2H). (1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methanol (Int-UU5-2, peak 2) was also isolated.

[0175] Step F: (S)-(1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methyl methanesulfonate (Int-UU6-1) (1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methanol (Int-UU5-1) (3.25 g, 14.24 mmol) in DCM (30 mL) was cooled to 0 °C and treated with triethylamine (7.94 mL, 57.0 mmol), followed by a solution of methanesulfonyl chloride (2.22 mL, 28.5 mmol) in DCM (2.2 mL). The reaction mixture was stirred and allowed to warm to room temperature over 3 h. The reaction mixture was purified by column chromatography on silica gel (0% to 100% EtOAc / hexanes) to give (S)-(1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methyl methanesulfonate (Int-UU6-1). MS (ESI): m / z [M+Na] + 329.

[0176] Step G: (R)-1-(1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)-N,N-dimethylmethanamine (Int-UU7-1) (S)-(1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methyl methanesulfonate (Int-UU6-1) (4.10 g, 13.4 mmol) and dimethylamine (2 M solution in THF) (33.5 mL, 66.9 mmol) were treated with potassium carbonate (3.70 g, 26.8 mmol). The flask was capped and heated at 50 °C for 24 h. The reaction mixture was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate. The combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude residue was purified by column chromatography on silica gel (120 g, 0% to 100% [1:3 EtOH / EtOAc] / hexanes) to give (R)-1-(1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)-N,N-dimethylmethanamine (Int-UU7-1). MS(ESI):m / z[M+H] + 256.

[0177] Step H: (R)-(1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methanol (Int-UU8-1) (R)-1-(1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)-N,N-dimethylmethanamine (Int-UU7-1) (2.96 g, 11.6 mmol) and Pd / C (10 wt %, wet support) (0.618 g, 0.580 mmol) in 2,2,2-trifluoroethanol (20.0 mL) were placed in a 100 mL recovery flask and stirred under hydrogen gas (1 atm, balloon) at room temperature for 20 h. The mixture was filtered through a pad of Celite, and the pad was washed with methanol (3 x 10 mL). The combined filtrate and washings were treated with 3 M HCl / methanol (12 mL, 36.0 mmol) and concentrated under reduced pressure to give a clear, colorless, viscous syrup. Diethyl ether (10 mL) was added, and the mixture was stirred to initiate precipitation. The mixture was concentrated under reduced pressure, treated with diethyl ether (10 mL), and sonicated for 1 minute. The diethyl ether was decanted off. The solid was dried under vacuum to give (R)-(1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methanol (Int-UU8-1). MS (ESI): m / z [M+H] +166. (S)-(1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methanol (Int-UU8-2) was synthesized by the same reaction procedure as above using Int-UU5-2 as an intermediate. MS(ESI): m / z [M+H] + 166.

[0178] (2R,7aS)-2-Fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methanol (Int-VV6) [ka]

[0179] Step A: Ethyl 2-(2-(chloromethyl)allyl)-5-oxopyrrolidine-2-carboxylate (Int-VV1) LiHMDS (1.00 M, 2.55 L) was added dropwise under N to a solution of ethyl 5-oxopyrrolidine-2-carboxylate (200 g, 1.27 mol) and 3-chloro-2-(chloromethyl)prop-1-ene (255 g, 2.04 mol, 236 mL) in THF (2.00 L) at −40° C. The mixture was stirred at 20° C. for 20 hours.

[0180] The reaction mixture was poured into saturated NH4Cl solution (1.00 L), and the pH of the mixture was adjusted to 6-7 with 1 N HCl. The resulting biphasic solution was extracted with EtOAc (3 x 500 mL). The organic phases were combined, washed with brine (600 mL), and concentrated under reduced pressure to give a crude residue. The crude material was purified by silica gel column chromatography (eluent: petroleum ether:ethyl acetate = 50:1 to 1:1 gradient) to give ethyl 2-(2-(chloromethyl)allyl)-5-oxopyrrolidine-2-carboxylate (Int-VV1). 1H NMR (400 MHz, CDCl3) δ 5.06 (br d, J = 17 Hz, 2H), 4.14 - 4.38 (m, 3H), 3.73 (br d, J = 16 Hz, 1H), 3.06 (br d, J = 16 Hz, 1H), 2.70 - 2.85 (m, 1H), 2.53 - 2.66 (m, 1H), 2.36 - 2.50 (m, 2H), 2.09 - 2.21 (m, 1H), 1.23 - 1.31 (m, 3H).

[0181] Step B: Ethyl 2-methylene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (Int-VV2) A solution of ethyl 2-(2-(chloromethyl)allyl)-5-oxopyrrolidine-2-carboxylate (Int-VV1) (500 g, 2.03 mol) in THF (500 mL) was added dropwise to a mixture of sodium hydride (97.7 g, 2.44 mol, 60.0% purity) in THF (3.00 L) at 0° C. under nitrogen. The reaction mixture was stirred at 70° C. under nitrogen for 12 hours. The reaction mixture was cooled and poured into saturated ammonium chloride solution (2.00 L) and stirred at 5° C. for 1 hour. The biphasic mixture was extracted with EtOAc (3×600 mL). The combined organic phases were washed with brine (2×500 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The crude residue was purified by silica gel column chromatography (eluent: petroleum ether:ethyl acetate=50:1 to 1:1) to give ethyl 2-methylene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (Int-VV2). 1H NMR (400 MHz, CDCl3) δ 4.96 - 5.13 (m, 2H), 4.27 (br d, J = 16 Hz, 1H), 4.19 (q, J = 7 Hz, 2H), 3.71 (br d, J = 16 Hz, 1H), 3.04 (d, J = 16 Hz, 1H), 2.69 - 2.83 (m, 1H), 2.59 (ddd, J = 2, 9, 13 Hz, 1H), 2.40 - 2.52 (m, 2H), 1.96 - 2.22 (m, 1H), 1.26 (t, J = 7 Hz, 3H).

[0182] Step C: Ethyl 2,5-dioxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (Int-VV3) Ozone (239 mmol) (0.5-1 m 3 1000kJ / h) was bubbled through a solution of ethyl 2-methylene-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (Int-VV2) (160 g, 765 mmol) in DCM (1.60 L) and MeOH (160 mL) at −70° C. for 9 hours. Nitrogen was bubbled through the reaction mixture to purge excess ozone. Dimethyl sulfide (76 g, 1.22 mol) was then added to the mixture at −70° C. The reaction mixture was stirred at 20° C. for 14 hours. The reaction mixture was concentrated under reduced pressure to give a residue. The crude residue was purified by silica gel column chromatography (eluent: petroleum ether:ethyl acetate = 50:1 to 1:1) to give ethyl 2,5-dioxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (Int-VV3). 1 H NMR (400 MHz, CDCl3) δ 4.22 (q, J = 7 Hz, 2H), 4.07 - 4.12 (m, 1H), 3.54 (dd, J = 1, 18 Hz, 1H), 2.92 - 3.03 (m, 2H), 2.74 - 2.88 (m, 1H), 2.42 - 2.51 (m, 2H), 2.12 - 2.23 (m, 1H), 1.27 (t, J = 7 Hz, 3H)

[0183] Step D: Ethyl 2-hydroxy-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (Int-VV4) To a solution of ethyl 2,5-dioxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (Int-VV3) (200 g, 947 mmol) in EtOH (2.00 L) at 0 °C under N was added NaBH (10.8 g, 284 mmol). The reaction mixture was stirred at 0 °C for 10 minutes. The reaction mixture was quenched by the addition of saturated NH Cl (50.0 mL) at 5 °C, and the mixture was stirred at 5 °C for 0.5 hours. The reaction mixture was concentrated under reduced pressure. The crude products from four identical reactions were then combined. The crude residue was purified by silica gel column chromatography (eluent: petroleum ether:ethyl acetate = 50:1 to 1:1) to give ethyl 2-hydroxy-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (Int-VV4). 1 H NMR (400 MHz, CDCl3) δ 4.54 - 4.70 (m, 1H), 4.16 - 4.31 (m, 2H), 3.93 (dd, J = 6.0, 13 Hz, 1H), 3.09 (d, J = 13 Hz, 1H), 2.75 - 2.90 (m, 1H), 2.39 - 2.63 (m, 4H), 2.01 - 2.13 (m, 1H), 1.83 (dd, J = 6, 14 Hz, 1H), 1.29 (t, J = 7 Hz, 3H).

[0184] Step E: Ethyl (2R,7aS)-2-fluoro-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (Int-VV5) To a solution of ethyl 2-hydroxy-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (Int-VV4) in DCM (600 mL) was added DAST (90.7 g, 563 mmol, 74.4 mL) dropwise at −70° C. under N. The reaction mixture was warmed to 20° C. and stirred for 16 h. The reaction was quenched by the addition of EtOH (50.0 mL) at 10° C., then diluted with water (300 mL) and extracted with DCM (2×200 mL). The combined organic phase was washed with brine (200 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure. The crude product was combined from six identical reactions and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 50:1 to 1:1). This material was further purified by preparative HPLC using HCl modifier (MeCN / water + 0.05% HCl modifier). The racemic mixture was resolved using chiral SFC (Daicel Chiralpak AS (50 mm × 250 mm, 10 μm; 0.1% NH4OH / EtOH) to give ethyl (2R,7aS)-2-fluoro-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (Int-VV5, peak 2). 1 H NMR (400 MHz, CDCl3): δ 5.16 - 5.43 (m, 1H), 4.14 - 4.27 (m, 3H), 3.06 - 3.26 (m, 1H), 2.57 - 2.85 (m, 3H), 2.38 - 2.50 (m, 1H), 2.07 - -2.30 (m, 2H), 1.28 (t, J = 7 Hz, 3H).

[0185] Step F: ((2R,7aS)-2-Fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methanol (Int-VV6) A solution of ethyl (2R,7aS)-2-fluoro-5-oxotetrahydro-1H-pyrrolidine-7a(5H)-carboxylate (Int-VV5) (82.0 g, 381 mmol) in THF (300 mL) was added to a mixture of LAH (21.7 g, 571 mmol) in THF (520 mL) at 0 °C under nitrogen. The reaction mixture was warmed to 70 °C and stirred for 3 h. The reaction mixture was cooled to 0 °C and quenched by the addition of NaSO·10H O at 0 °C under nitrogen. The reaction mixture was stirred at 20 °C for 0.5 h and then filtered. The filter cake was washed with EtOAc (5 x 600 mL), and the filtrate was dried over anhydrous MgSO. The mixture was filtered, and the filtrate was concentrated under reduced pressure to give a residue. The crude residue was purified by silica gel column chromatography (SiO 2 , DCM:methanol=100:1 to 10:1) to give ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolidin-7a(5H)-yl)methanol (Int-VV6). 1 H NMR (400 MHz, CDCl3): δ 5.06 - 5.34 (m, 1H), 3.25 (s, 2H), 3.08 - 3.23 (m, 3H), 2.85 - 3.08 (m, 2H), 2.00 - 2.12 (m, 2H), 1.74 - 1.93 (m, 4H).

[0186] 4-Bromo-6-methyl-1-(tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)-1H-indazole (Int-HHH5) [ka]

[0187] Step A: 1-Bromo-5-fluoro-2-iodo-3-methylbenzene (Int-HHH1) 2-Bromo-4-fluoro-6-methylaniline (200 g, 0.983 mol) was dissolved in MeCN (800 mL). The resulting mixture was cooled to 0 °C. Concentrated HCl (12 M, 245 mL) was added to the reaction mixture while maintaining the reaction temperature at 0 °C. A solution of NaNO (81.1 g, 1.18 mol equivalents) in water (400 mL) was added dropwise to the reaction mixture while maintaining the reaction temperature at 0 °C. The resulting mixture was stirred at 0 °C for 0.5 h. Next, a solution of KI (195 g, 1.18 mol) in water (400 mL) was added dropwise to the reaction mixture at 0 °C. The resulting mixture was allowed to warm to room temperature and stirred at 20 °C for 12 h. This reaction was repeated one more time using the conditions described above. The two reaction batches were combined. The product mixture was adjusted to pH 8-9 with aqueous NaOH, and the aqueous phase was extracted with EtOAc (2 x 2.00 L). The organic phase was dried over Na2SO4, filtered, and concentrated, and the resulting residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 1:0 to 0:1) to give 1-bromo-5-fluoro-2-iodo-3-methylbenzene (Int-HHH1). 1 H NMR (400 MHz, CDCl3) δ 7.27 - 7.22 (m, 1H), 6.95 (dd, J = 2.4, 8.8 Hz, 1H), 2.56 (s, 3H).

[0188] Step B: 1-Bromo-5-fluoro-3-methyl-2-(trifluoromethyl)benzene (Int-HHH2) 1-Bromo-5-fluoro-2-iodo-3-methylbenzene (Int-HHH1) (100 g, 0.317 mol) was dissolved in DMF (1.50 L). To this mixture was added CuI (514 g, 2.70 mol) and methyl 2,2-difluoro-2-(fluorosulfonyl)acetate (518 g, 2.70 mol) at 25°C. The reaction mixture was heated and stirred at 60°C for 12 hours. This reaction was repeated three more times using the above conditions. The four reaction batches were combined and quenched with water (24 L). The mixture was extracted with petroleum ether (2 x 8.00 L). The combined organic phase was washed with brine (2 x 4 L) and dried over Na2SO4. The dried solution was filtered and the filtrate was concentrated under reduced pressure to give a crude material containing 1-bromo-5-fluoro-3-methyl-2-(trifluoromethyl)benzene (Int-HHH2), which was used directly in the next step without purification.

[0189] Step C: 2-Bromo-6-fluoro-4-methyl-3-(trifluoromethyl)benzaldehyde (Int-HHH3) 1-Bromo-5-fluoro-3-methyl-2-(trifluoromethyl)benzene (Int-HHH2) (100 g, 0.382 mol) was dissolved in 2-MeTHF (500 mL). The reaction mixture was cooled to −65°C. To the mixture at −65°C was added 2 M LDA solution (213 mL, 426 mmol). The reaction mixture was stirred at −65°C for 0.5 h. To the mixture was added DMF (31.2 g, 0.420 mol) dropwise at −65°C. The reaction mixture was stirred at −65°C for 2 h. This reaction was repeated two more times using the above conditions. The three reaction batches were combined. The pH of the reaction mixture was adjusted to 3–4 using 1 M HCl, and the aqueous phase was extracted with 2-MeTHF (2×500 mL). The organic phase was dried over Na2SO4, filtered and concentrated to give 2-bromo-6-fluoro-4-methyl-3-(trifluoromethyl)benzaldehyde (Int-HHH3), which was used in the next step without further purification.

[0190] Step D: 4-Bromo-6-methyl-5-(trifluoromethyl)-1H-indazole (Int-HHH4) 2-Bromo-6-fluoro-4-methyl-3-(trifluoromethyl)benzaldehyde (Int-HHH3) (100 g, 0.351 mol) was dissolved in THF (800 mL). To this mixture was added N2H4·H2O (53.7 g, 1.05 mol) at 25 °C. The mixture was heated and stirred at 60 °C for 2 h. The product mixture was quenched with water (400 mL) and extracted with EtOAc (2 x 200 mL). The combined organic phases were washed with brine (200 mL) and dried over Na2SO4. The dried solution was filtered, and the filtrate was concentrated under reduced pressure to give a residue. This reaction was repeated two more times using the above conditions. The three reaction batches were combined. The resulting residue was triturated with DCM (100 mL) at 15 °C for 2 h. The solid was collected by filtration to give 4-bromo-6-methyl-5-(trifluoromethyl)-1H-indazole (Int-HHH4). 1 H NMR (400 MHz, CDCl3) δ 10.61 - 10.20 (m, 1H), 8.20 (d, J = 0.8 Hz, 1H), 7.34 (d, J = 0.6 Hz, 1H), 2.67 - 2.63 (m, 3H).

[0191] Step E: 4-Bromo-6-methyl-1-(tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)-1H-indazole (Int-HHH5) 4-Bromo-6-methyl-5-(trifluoromethyl)-1H-indazole (Int-HHH4) (60.0 g, 0.215 mol) was dissolved in DCM (240 mL) and MeCN (240 mL). To the mixture was added DHP (21.7 g, 0.258 mol) and TsOH·HO (8.18 g, 0.043 mol) at 20 °C. The reaction mixture was stirred at 20 °C for 12 h. Water (200 mL) was added to the product mixture. The resulting mixture was extracted with DCM (2 x 200 mL). The combined organic phase was washed with brine (200 mL) and dried over Na2SO4. The dried solution was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 1:0 to 0:1) to give 4-bromo-6-methyl-1-(tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)-1H-indazole (Int-HHH5). 1 H NMR (400 MHz, CDCl3- d) δ 8.12 (s, 1H), 7.44 (s, 1H), 5.69 (dd, J = 3, 9 Hz, 1H), 4.09 - 3.94 (m, 1H), 3.81 - 3.69 (m, 1H), 2.69 - 2.63 (m, 3H), 2.56 - 2.43 (m, 1H), 2.19 - 2.14 (m, 1H), 2.12 - 2.04 (m, 1H), 1.87 - 1.73 (m, 2H), 1.71 - 1.63 (m, 1H).

[0192] 1,8-Dibromo-3-(methoxymethoxy)naphthalene (Int-W002) [ka]

[0193] Step A: 2,4,5-Tribromonaphthalen-1-amine (Int-W001) To a solution of 5-bromonaphthalen-1-amine (63 g, 280 mmol) in DMA (1260 mL) was added NBS (106 g) at 0 °C. The mixture was warmed to room temperature and stirred for 3 h. The reaction mixture was diluted with a solution of NaSO (75 g) in HO (380 mL) and NaHCO (24 g) in HO (1100 mL) and stirred for 1 h. The precipitate was collected by filtration and washed with water to give 2,4,5-tribromonaphthalen-1-amine (Int-W001) (97 g) as a purple solid. 1 H NMR (400 MHz, CDCl3) δ 7.99 (s, 1H), 7.97-7.95 (1H, m), 7.86-7.84 (1H, m), 7.31-7.29 (1H, m), 4.65 (2H, brs). LCMS (ESI): m / z [M+H] + 379.9.

[0194] Step B: 1,8-Dibromo-3-(methoxymethoxy)naphthalene (Int-W002) To a suspension of 2,4,5-tribromonaphthalen-1-amine (Int-W001) (60 g, 160 mmol) in AcOH (780 mL) and propionic acid (300 mL) at 0 °C, NaNO (11 g) was added portionwise and the reaction mixture was stirred for 20 min. The reaction mixture was diluted with HO (1800 mL) at 0 °C and stirred for 1 h. The resulting slurry was filtered and the solid was washed with HO to give 4,5-dibromo-2-hydroxynaphthalene-1-diazonium, which was used without further purification.

[0195] To a suspension of 4,5-dibromo-2-hydroxynaphthalene-1-diazonium in EtOH (1600 mL) at 0 °C, NaBH (15 g) was added portionwise, and the reaction mixture was stirred for 30 min. The mixture was warmed to room temperature and stirred overnight. The reaction mixture was cooled to 0 °C and diluted with water (1500 mL) and 5 M aqueous HCl (79 mL). The mixture was evaporated to remove EtOH and extracted with CHCl. ​​The combined organic phases were washed with brine, dried over NaSO, and evaporated under reduced pressure to give 4,5-dibromonaphthalen-2-ol, which was used without further purification.

[0196] To a solution of 4,5-dibromonaphthalen-2-ol in CHCl (900 mL) was added i-PrNEt (170 mL) and MOMCl (36 m) at 0 °C. After stirring at room temperature for 1 h, the reaction mixture was diluted with EtOAc and saturated NaHCO. The organic phase was separated, and the aqueous layer was extracted with EtOAc. The combined organic phases were washed with brine, dried over NaSO, and evaporated. The resulting residue was purified by silica gel column chromatography to give 1,8-dibromo-3-(methoxymethoxy)naphthalene (Int-W002) (17.4 g). 1 H NMR (400 MHz, CHLOROFORM-d) δ = 7.82 (dd, J = 1.3, 7.4 Hz, 1H), 7.75 - 7.71 (m, 2H), 7.41 (d, J = 2.6 Hz, 1H), 7.26 - 7.20 (m, J = 8.0, 8.0 Hz, 1H), 5.29 (s, 2H), 3.53 (s, 3H).

[0197] Example 1: (4-(azepan-1-yl)-2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 1) [ka]

[0198] To a mixture of (2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-hydroxy-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone dihydrochloride (Int-L004) (15.0 mg, 0.0237 mmol) and p-toluenesulfonyl chloride (9.9 mg, 0.0521 mmol) in DMF (0.3 mL) was added N,N-diisopropylethylamine (0.032 mL, 0.189 mmol), and the mixture was stirred at room temperature for 0.5 h. Hexamethyleneimine (0.0107 mL, 0.0947 mmol) was added, and the mixture was stirred at 70 °C for 1 h. Next, aqueous NaOH (2 M, 0.4 mL) was added to the mixture, and the mixture was stirred at 70 °C for an additional 1 h. The mixture was cooled to room temperature, diluted with DMSO (0.5 mL), and the product mixture was purified by reverse-phase HPLC (MeCN / water with 0.1% formic acid) to give (4-(azepan-1-yl)-2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 1). ESI-MS m / z [M+H] + 642. 1H NMR (400 MHz, DMSO-d6) δ 10.2 (s, 1H), 8.03-7.98 (m, 1H), 7.87-7.82 (m, 1H), 7.29-7.26 (m, 1H), 7.16-7.10 (m, 2H), 4.98 (br s, 1H), 4.70-4.45 (m, 2H), 4.19-4.00 (m, 4H), 3.70-3.62 (m, 1H), 2.35-2.18 (m, 2H), 2.16 (s, 3H), 2.12 (s, 3H), 1.83-1.70 (m, 2H), 1.60-1.34 (m, 8H), 0.59-0.56 (m, 2H), 0.54-0.51 (m, 2H).

[0199] The compounds in the following table were synthesized using (2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-hydroxy-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone dihydrochloride (Int-L004) or (1-((dimethylamino)methyl)cyclopropyl)methanol and the corresponding amine via a route similar to that in Example 1.

[0200] Table 1 [Table 1] TIFF2024521979000037.tif255168TIFF2024521979000038.tif255149TIFF2024521979000039.tif255167TIFF2024521979000040.tif255166TIFF202 4521979000041.tif255153TIFF2024521979000042.tif255166TIFF2024521979000043.tif255162TIFF2024521979000044.tif255157TIFF2024521979 000045.tif255167TIFF2024521979000046.tif255167TIFF2024521979000047.tif255162TIFF2024521979000048.tif255163TIFF2024521979000049. tif255167TIFF2024521979000050.tif255148TIFF2024521979000051.tif255166TIFF2024521979000052.tif255153TIFF2024521979000053.tif25535

[0201] Example 82: (4-(3-amino-5-hydroxypiperidin-1-yl)-2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 82) [ka]

[0202] To a solution of tert-butyl (1-(2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-6-(3-hydroxy-8-iodo-1-naphthoyl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)-5-hydroxypiperidin-3-yl)carbamate (Example 67) (30 mg, 0.0395 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (5 mL) and the mixture was stirred for 1 hour. The trifluoroacetic acid was removed under reduced pressure, and the residue was purified by flash NH-silica gel chromatography (0% to 50%, methanol gradient / ethyl acetate) to give (4-(3-amino-5-hydroxypiperidin-1-yl)-2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 82). ESI-MS m / z [M+H] + 659.

[0203] Example 83: N-(1-(2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-6-(3-hydroxy-8-iodo-1-naphthoyl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)-5-hydroxypiperidin-3-yl)-1-methyl-1H-pyrazole-5-carboxamide (Example 83) [ka]

[0204] A solution of 1-methyl-1H-pyrazole-5-carboxylic acid (3.0 mg, 0.0237 mmol), 1-hydroxybenzotriazole monohydrate (2.9 mg, 0.0189 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.6 mg, 0.0189 mmol), and N,N-diisopropylethylamine (0.006 mL, 0.0316 mmol) in N,N-dimethylformamide was added. To the mixture in 1H-dichloromethane (0.3 mL) was added (4-(3-amino-5-hydroxypiperidin-1-yl)-2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 82) (10 mg, 0.015 mmol) and the mixture was stirred at room temperature for 5 hours. The mixture was diluted with DMSO (0.6 mL) and the product mixture was purified by reverse-phase HPLC (MeCN / water with 0.1% formic acid) to give N-(1-(2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-6-(3-hydroxy-8-iodo-1-naphthoyl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)-5-hydroxypiperidin-3-yl)-1-methyl-1H-pyrazole-5-carboxamide (Example 83). ESI-MS m / z [M+H] + 767. 1 H NMR (400 MHz, DMSO-d6) δ 10.3 (s, 1H), 8.49-8.13 (m, 1H), 8.04-7.96 (m, 1H), 7.89-7.78 (m, 1H), 7.47-7.39 (m, 1H), 7.30-7.20 (m, 1H), 7.18-7.05 (m, 2H), 6.84-6.66 (m, 1H), 5.15-4.52 (m, 2H), 4.28-3.65 (m, 7H), 3.64-3.12 (m, 10H) 2.25-2.18 (m, 1H), 2.16 (s, 3H), 2.11 (s, 3H), 1.93-1.43 (m, 1H), 0.61-0.48 (m, 2H), 0.40-0.30 (m, 2H)

[0205] Example 84-1: (2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 84-1) [ka]

[0206] Step A: tert-Butyl 2-chloro-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-84a) To a stirred solution of tert-butyl 2,4-dichloro-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (3.88 g, 13.4 mol) and 2-methylazepane hydrochloride (2.00 g, 13.4 mol) in DME (30 mL) was added N,N-diisopropylethylamine (6.82 mL, 40.1 mmol), and the mixture was warmed to 50 °C and stirred for 5 h. The mixture was cooled to room temperature and diluted with ethyl acetate. The diluted mixture was washed with water, and the mixture was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 40%, ethyl acetate gradient in hexane) to give tert-butyl 2-chloro-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-84a) (3.52 g). ESI-MS m / z [M+H] + 367, 369.

[0207] Step B: tert-Butyl 2-chloro-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-84a-1 Peak 1 and Int-84a-2 Peak 2) by chiral HPLC Racemic tert-butyl 2-chloro-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-84a) was separated by chiral HPLC (column: Chiral Art SB 4.6 mmφ (YMC), hexane / EtOH 0.1% diethylamine) to give tert-butyl 2-chloro-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-84a-1, first elution): ESI-MS m / z [M+H] +367, 369 and tert-butyl 2-chloro-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-84a-2, second elution): ESI-MS m / z [M+H] + I got 367 and 369.

[0208] Step C: N,N-Dimethyl-1-(1-(((4-(2-methylazepan-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanamine (Int-84b) To a mixture of tert-butyl 2-chloro-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-84a-2 peak 2, second elution) (360 mg, 0.981 mmol), (1-((dimethylamino)methyl)cyclopropyl)methanol (254 mg, 1.96 mmol), and dichloro[9,9-dimethyl-4,5-bis(diphenylphosphino)xanthene]palladium(II) (74 mg, 0.098 mmol) in toluene (10 mL) was added cesium carbonate (959 mg, 2.94 mmol), and the mixture was warmed to 130 °C and stirred for 3 h. The mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 25%, methanol gradient / chloroform) to give the coupled product. To a solution of the coupling product in dichloromethane (2 mL) was added trifluoroacetic acid (2 mL), and the mixture was stirred for 1 h. Trifluoroacetic acid was removed under reduced pressure, and the residue was purified by flash NH-silica gel chromatography (0% to 50%, methanol gradient / ethyl acetate) to give N,N-dimethyl-1-(1-(((4-(2-methylazepan-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanamine (Int-84b) (180 mg). ESI-MS m / z [M+H] + 360.

[0209] Step D: Example 84-1: (2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 84) To a mixture of 3-hydroxy-8-iodo-1-naphthoic acid (Int-T002) (105 mg, 0.334 mmol), 1-hydroxybenzotriazole monohydrate (51 mg, 0.334 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (64 g, 0.334 mmol), and N,N-diisopropylethylamine (0.14 mL, 0.834 mmol) in N,N-dimethylformamide (1 mL) was added N,N-dimethyl-1-(1-(((4-(2-methylazepan-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanamine (Int-84b) (100 mg, 0.278 mmol), and the mixture was stirred at 40 °C for 2 hours. The mixture was diluted with ethyl acetate, washed with water, and concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (0% to 60%, methanol gradient / ethyl acetate) to give (2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 84-1). ESI-MS m / z [M+H] + 656. 1 H NMR (400 MHz, DMSO-d6) δ 10.3 (s, 1H), 8.02-7.98 (m, 1H), 7.88-7.80 (m, 1H), 7.29-7.24 (m, 1H), 7.19-7.09 (m, 2H), 5.15-5.04 (m, 1H), 4.96-4.88 (m, 1H), 4.70-4.38 (m, 1H), 4.22-4.00 (m, 5H), 3.21-2.80 (m, 3H), 2.26-2.10 (m, 1H), 2.15 (s, 3H), 2.11 (s, 3H), 1.85-0.90 (m, 8H), 0.59-0.50 (m, 2H), 0.39-0.31 (m, 2H).

[0210] Example 84-2: (3-hydroxy-8-iodonaphthalen-1-yl)(2-((1-(hydroxymethyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 84-2) [ka]

[0211] Step A: (1-(((4-(2-methylazepan-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanol (Int-84c-1) To a mixture of tert-butyl 2-chloro-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-84a) (1.50 g, 4.09 mmol), 1,1-bis(hydroxymethyl)cyclopropane (1.25 g, 12.3 mmol), and RuPhosPdG (171 mg, 0.204 mmol) in 1,4-dioxane (15 mL) was added cesium carbonate (4.00 g, 12.3 mmol), and the mixture was warmed to 110 °C and stirred for 15 min. The mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (40% to 80%, ethyl acetate gradient / hexanes) to give the coupled product. To a solution of the coupling product in dichloromethane (5 mL) was added trifluoroacetic acid (5 mL), and the mixture was stirred for 1 h. Trifluoroacetic acid was removed under reduced pressure, and the residue was purified by flash NH-silica gel chromatography (0% to 50%, methanol gradient / ethyl acetate) to give (1-(((4-(2-methylazepan-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanol (Int-84c-1) (1.36 g). ESI-MS m / z [M+H] + 333.

[0212] Step B: (3-Hydroxy-8-iodonaphthalen-1-yl)(2-((1-(hydroxymethyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 84-2) To a mixture of 3-hydroxy-8-iodo-1-naphthoic acid (Int-T002) (1.54 g, 4.91 mmol), 1-hydroxybenzotriazole monohydrate (751 mg, 4.91 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (941 g, 4.91 mmol), and N,N-diisopropylethylamine (2.09 mL, 12.3 mmol) in N,N-dimethylformamide (15 mL) was added (1-(((4-(2-methylazepan-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanol (Int-84c-1) (1.36 g, 4.09 mmol), and the mixture was stirred at room temperature for 2 hours. The mixture was diluted with ethyl acetate, washed with water, and concentrated under reduced pressure. The residue was suspended in ethyl acetate (10 mL), and the solid was filtered and dried under vacuum to give (3-hydroxy-8-iodonaphthalen-1-yl)(2-((1-(hydroxymethyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 84-2) (1.45 g). ESI-MS m / z [M+H] + 629.

[0213] Example 85: (3-hydroxy-8-iodonaphthalen-1-yl)(2-((1-(((R)-3-methoxypiperidin-1-yl)methyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 85) [ka]

[0214] To a mixture of (3-hydroxy-8-iodonaphthalen-1-yl)(2-((1-(hydroxymethyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 84-2) (20.0 mg, 0.0318 mmol) and N,N-diisopropylethylamine (27 mL, 0.159 mmol) in DMF (0.3 mL) was added ethanesulfonyl chloride (12.3 mg, 0.0955 mmol) at 0 ° C., and the mixture was stirred at 0 ° C. for 0.5 hours. (R)-3-Methoxypiperidine (18.3 mg, 0.159 mmol) was added to the mixture, and the mixture was stirred at 70 ° C. for 1 hour. Methanol (0.1 mL) and aqueous NaOH (2 M, 0.2 mL) were then added to the mixture, and the mixture was stirred at 70° C. for an additional 0.5 h. The mixture was cooled to room temperature, diluted with DMSO (0.6 mL), and the product mixture was purified by reverse-phase HPLC (MeCN / water with 0.1% formic acid) to give (3-hydroxy-8-iodonaphthalen-1-yl)(2-((1-(((R)-3-methoxypiperidin-1-yl)methyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 85). ESI-MS m / z [M+H] + 726. 1 H NMR (400 MHz, DMSO-d6) δ 10.2 (s, 1H), 8.04-7.98 (m, 1H) 7.88-7.82 (m, 1H), 7.30-7.26 (m, 1H), 7.19-7.09 (m, 2H), 5.15-5.03 (m, 1H), 4.98-4.87 (m, 1H), 4.70-4.35 (m, 2H) 4.21-3.99 (m, 5H), 3.23 (s, 1.5H), 3.28 (s, 1.5H), 3.15-2.78 (m, 2H), 2.31-2.11(m, 2H), 2.00-0.95(m, 24H), 0.61-0.51 (m, 2H), 0.40-0.31 (m, 2H).

[0215] Using Example 84-2 and the corresponding amine, the compounds in the following table were synthesized by the same route as in Example 85.

[0216] Table 2 [Table 2] TIFF2024521979000060.tif255110

[0217] Example 84-2a: (R)-(3-hydroxy-8-iodonaphthalen-1-yl)(2-((1-(hydroxymethyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 84-2a) [ka]

[0218] Step A: tert-Butyl (R)-2-chloro-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-84a-3) To a stirred solution of tert-butyl 2,4-dichloro-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (2.13 g, 7.35 mol) and (R)-2-methylazepane hydrochloride (1.00 g, 6.68 mol) in DME (20 mL) was added N,N-diisopropylethylamine (3.41 mL, 20.0 mmol), and the mixture was warmed to 60° C. and stirred for 4 hours. The mixture was cooled to room temperature and diluted with ethyl acetate. The diluted mixture was washed with water, and the mixture was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 40%, ethyl acetate gradient / hexanes) to give tert-butyl (R)-2-chloro-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-84a-3) (1.92 g). ESI-MS m / z [M+H] + 367, 369.

[0219] Step B: (R)-(1-(((4-(2-methylazepan-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanol (Int-84c-2) To a mixture of tert-butyl (R)-2-chloro-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-84a-3) (920 mg, 2.51 mmol), 1,1-bis(hydroxymethyl)cyclopropane (768 mg, 7.52 mmol), and RuPhosPdG3 (52.4 mg, 0.0627 mmol) in 1,4-dioxane (15 mL) was added cesium carbonate (2.45 g, 7.52 mmol), and the mixture was warmed to 110 °C and stirred for 30 min. The mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (40% to 80%, ethyl acetate gradient / hexanes) to give the coupled product. To a solution of the coupling product in dichloromethane (5 mL) was added trifluoroacetic acid (5 mL), and the mixture was stirred for 1 h. Trifluoroacetic acid was removed under reduced pressure, and the residue was purified by flash NH-silica gel chromatography (0% to 50%, methanol gradient / ethyl acetate) to give (R)-(1-(((4-(2-methylazepan-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanol (Int-84c-2) (500 mg). ESI-MS m / z [M+H] + 333.

[0220] Step C: (R)-(3-hydroxy-8-iodonaphthalen-1-yl)(2-((1-(hydroxymethyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 84-2a) To a mixture of 3-hydroxy-8-iodo-1-naphthoic acid (Int-T002) (471 mg, 1.50 mmol), 1-hydroxybenzotriazole monohydrate (230 mg, 1.50 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (288 mg, 1.50 mmol), and N,N-diisopropylethylamine (0.64 mL, 3.75 mmol) in N,N-dimethylformamide (5 mL) was added (R)-(1-(((4-(2-methylazepan-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanol (Int-84c-2) (416 mg, 1.25 mmol), and the mixture was stirred at room temperature for 5 hours. The mixture was diluted with ethyl acetate, washed with water, and concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (0% to 100%, ethyl acetate gradient / hexane) to give (R)-(3-hydroxy-8-iodonaphthalen-1-yl)(2-((1-(hydroxymethyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 84-2a) (580 mg). ESI-MS m / z [M+H] + 629.

[0221] Example 93: (R)-(3-hydroxy-8-iodonaphthalen-1-yl)(2-((1-((4-methoxypiperidin-1-yl)methyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 93) [ka]

[0222] Example 93 was synthesized using (R)-(3-hydroxy-8-iodonaphthalen-1-yl)(2-((1-(hydroxymethyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 84-2a) and 4-methoxypiperidine by the same procedure as Example 85. ESI-MS m / z [M+H]+ 726. 1H NMR (400 MHz, DMSO-d6) δ 10.3 (s, 1H), 8.03-7.98 (m, 1H), 7.87-7.82 (m, 1H), 7.29-7.25 (m, 1H), 7.19-7.09 (m, 2H), 5.18-5.04 (m, 1H), 4.96-4.88 (m, 1H), 4.69-4.35 (m, 2H), 4.22-4.00 (m, 4H), 3.21 (s, 1.5H), 3.19 (s, 1.5H), 2.80-2.65 (m, 2H), 2.38-2.15 (m, 2H), 2.10-0.95 (m, 19H), 0.59-0.50 (m, 2H), 0.39-0.30 (m, 2H).

[0223] Example 94: (R)-(2-((1-((1,4-oxazepan-4-yl)methyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 94) [ka]

[0224] Example 94 was synthesized using (R)-(3-hydroxy-8-iodonaphthalen-1-yl)(2-((1-(hydroxymethyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 84-2A) and 1,4-oxyazepane by the same procedure as Example 85. ESI-MS m / z [M+H]+ 712.

[0225] Example 95: (R)-(4-(3-hydroxy-3-methylpiperidin-1-yl)-2-((1-(hydroxymethyl)cyclopropyl)methoxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 95) [ka]

[0226] Step A: tert-Butyl (R)-2-chloro-4-(3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-95a) To a solution of tert-butyl 2,4-dichloro-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (900 mg, 3.10 mmol) and (R)-3-methylpiperidin-3-ol hydrochloride (470 mg, 3.10 mmol) in N,N-dimethylacetamide (5.0 mL) was added N,N-diisopropylethylamine (1.62 mL, 9.31 mmol) at room temperature. The mixture was stirred at room temperature for 40 minutes and then quenched by the addition of HO. The reaction mixture was extracted with EtOAc, washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 100%, EtOAc gradient / hexanes) to give tert-butyl (R)-2-chloro-4-(3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate ( Int-95a ) (1.12 g) was obtained. ESI-MS m / z [M+H] + 369, 371.

[0227] Step B: (R)-1-(2-((1-(hydroxymethyl)cyclopropyl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (Int-95b) tert-Butyl (R)-2-chloro-4-(3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate ( Int-95aTo a solution of RuPhosPdG3 (50.0 mg, 0.136 mmol), cyclopropane-1,1-diyldimethanol (41.5 mg, 0.407 mmol), and RuPhosPdG3 (3.40 mg, 0.00407 mmol) in 1,4-dioxane (0.7 mL) was added cesium carbonate (132 mg, 0.407 mmol) at room temperature. After stirring the mixture at 100 °C for 1 h, the reaction mixture was cooled to room temperature, filtered, and concentrated under reduced pressure. The residue was dissolved in EtOAc, washed with HO and brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to give the coupling product. To a solution of the coupling product in dichloromethane (2.6 mL) was added trifluoroacetic acid (1.0 mL). After stirring the mixture at room temperature for 3 h, the reaction mixture was concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (0% to 50%, MeOH gradient / EtOAc) to give (R)-1-(2-((1-(hydroxymethyl)cyclopropyl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol ( Int-95b ) (37.5 mg) was obtained. ESI-MS m / z [M+H] + 335.

[0228] Step C: Example 95 (R)-(4-(3-hydroxy-3-methylpiperidin-1-yl)-2-((1-(hydroxymethyl)cyclopropyl)methoxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 95) To a solution of (R)-1-(2-((1-(hydroxymethyl)cyclopropyl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (Int-95b) (37.5 mg, 0.112 mmol), 3-hydroxy-8-iodo-1-naphthoic acid (Int-T002) (38.7 mg, 0.123 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (23.6 mg, 0.123 mmol), and 1-hydroxybenzotriazole hydrate (18.9 mg, 0.123 mmol) in DMF (1.2 mL) was added N,N-diisopropylethylamine (0.0977 mL, 0.561 mmol) at room temperature. The mixture was stirred at room temperature for 15 h and then quenched by the addition of HO. The reaction mixture was extracted with EtOAc, washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (0% to 50%, MeOH gradient / EtOAc) and reverse-phase HPLC (MeCN / water + 0.1% formic acid) to give (R)-(4-(3-hydroxy-3-methylpiperidin-1-yl)-2-((1-(hydroxymethyl)cyclopropyl)methoxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 95). ESI-MS m / z [M+H] + 631. 1 H NMR (400 MHz, DMSO-d6) δ 10.3-10.2 (m, 1H), 8.02-7.96 (m, 1H), 7.87-7.80 (m, 1H), 7.29-7.24 (m, 1H), 7.16-7.08 (m, 2H), 5.06-4.83 (m, 1H), 4.68-4.34 (m, 4H), 4.26-3.96 (m, 4H), 3.90-3.64 (m, 1H), 3.57-3.44 (m, 1H), 3.25-2.96 (m, 2H), 1.85-1.38 (m, 4H), 1.13 (s, 3H), 0.80 (d, J = 10.6 Hz, 1H), 0.52-0.39 (m, 4H).

[0229] Example 96: (R)-(4-(3-hydroxy-3-methylpiperidin-1-yl)-2-((1-(piperidin-1-ylmethyl)cyclopropyl)methoxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 96) [ka]

[0230] To a solution of (R)-(4-(3-hydroxy-3-methylpiperidin-1-yl)-2-((1-(hydroxymethyl)cyclopropyl)methoxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 95) (10.0 mg, 0.0159 mmol) in N,N-dimethylacetamide (0.3 mL) was added ethanesulfonyl chloride (0.00333 mL, 0.0352 mmol) and triethylamine (0.0111 mL, 0.0793 mmol) at 5° C. The mixture was stirred at 5° C. for 1 hour, and then piperidine (0.00783 mL, 0.0793 mmol) was added to the reaction mixture. After stirring the mixture at 75° C. for 2 hours, 2.0 M aqueous NaOH (0.2 mL) was added to the mixture, and the mixture was stirred at 75° C. for 30 minutes. The reaction mixture was cooled to room temperature, diluted with DMSO (0.5 mL), and purified by reverse-phase HPLC (MeCN / water with 0.1% formic acid) to give (R)-(4-(3-hydroxy-3-methylpiperidin-1-yl)-2-((1-(piperidin-1-ylmethyl)cyclopropyl)methoxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 96). ESI-MS m / z [M+H] + 698. 1H NMR (400 MHz, DMSO-d6) δ 10.3 (s, 1H), 8.02-7.96 (m, 1H), 7.87-7.80 (m, 1H), 7.29-7.24 (m, 1H), 7.16-7.08 (m, 2H), 5.06-4.82 (m, 1H), 4.70-4.33 (m, 3H), 4.19-3.96 (m,4H), 3.87-3.65 (m, 1H), 2.39-2.13 (m, 6H), 1.84-1.25 (m, 11H), 1.13 (s, 3H), 0.80 (d, J = 10.4Hz, 1H), 0.60-0.46 (m, 2H), 0.38-0.27 (m, 2H).

[0231] Using Example 95 and the corresponding amine, the compounds in the following table were synthesized by a similar route to Example 96.

[0232] Table 3 [Table 3] TIFF2024521979000067.tif255153TIFF2024521979000068.tif255116

[0233] (R)-(3-Hydroxy-8-iodonaphthalen-1-yl)(2-((1-(hydroxymethyl)cyclopropyl)methoxy)-4-(3-methyl-3-((trimethylsilyl)oxy)piperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Int-108e) [ka]

[0234] Step A: Benzyl (R)-2-chloro-4-(3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-108a) To a solution of benzyl 2,4-dichloro-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (300 mg, 0.925 mmol) and (R)-3-methylpiperidin-3-ol hydrochloride (140 mg, 0.925 mmol) in N,N-dimethylacetamide (5.0 mL) was added N,N-diisopropylethylamine (0.450 mL, 2.58 mmol) at room temperature. The mixture was stirred at room temperature for 30 minutes and then quenched by the addition of HO. The reaction mixture was extracted with EtOAc, washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 50%, EtOAc gradient / hexanes) to give benzyl (R)-2-chloro-4-(3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-108a) (357 mg). ESI-MS m / z [M+H] + 403, 405.

[0235] Step B: Benzyl (R)-2-chloro-4-(3-methyl-3-((trimethylsilyl)oxy)piperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-108b) To a solution of benzyl (R)-2-chloro-4-(3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-108a) (357 mg, 0.885 mmol) in DMF (3.0 mL) was added imidazole (121 mg, 1.77 mmol) and chlorotrimethylsilane (0.340 mL, 2.68 mmol) at room temperature. The mixture was stirred at room temperature for 15 h and then quenched by the addition of HO. The reaction mixture was extracted with EtOAc, washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 40%, EtOAc gradient / hexanes) to give benzyl (R)-2-chloro-4-(3-methyl-3-((trimethylsilyl)oxy)piperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-108b) (375 mg). ESI-MS m / z [M+H] + 475, 477.

[0236] Step C: (R)-(1-(((4-(3-methyl-3-((trimethylsilyl)oxy)piperidin-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanol (Int-108c) and (R)-(1-(((4-(3-methyl-3-((trimethylsilyl)oxy)piperidin-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl acetate (Int-108d) To a solution of benzyl (R)-2-chloro-4-(3-methyl-3-((trimethylsilyl)oxy)piperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-108b) (375 mg, 0.788 mmol), cyclopropane-1,1-diyldimethanol (242 mg, 2.36 mmol), and RuPhosPdG3 (19.8 mg, 0.0236 mmol) in 1,4-dioxane (5.0 mL) was added cesium carbonate (771 mg, 2.36 mmol) at room temperature. After stirring the mixture at 100 °C for 1 h, the reaction mixture was cooled to room temperature, diluted with EtOAc, filtered, and concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (50% to 100%, EtOAc gradient / hexanes) to give the coupled product.

[0237] To a solution of the coupled product in ethanol (20 mL) was added Pd(OH) / C (277 mg, 0.394 mmol). The reaction mixture was purged with H and evacuated three times and stirred at room temperature for 1 h. The reaction mixture was filtered and concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (0% to 30%, MeOH gradient / EtOAc) to give (R)-(1-(((4-(3-methyl-3-((trimethylsilyl)oxy)piperidin-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanol (Int-108c) (40.5 mg): ESI-MS m / z [M+H] + 407 and (R)-(1-(((4-(3-methyl-3-((trimethylsilyl)oxy)piperidin-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl acetate (Int-108d) (105 mg): ESI-MS m / z [M+H] + I got 449.

[0238] Step D: (R)-(3-Hydroxy-8-iodonaphthalen-1-yl)(2-((1-(hydroxymethyl)cyclopropyl)methoxy)-4-(3-methyl-3-((trimethylsilyl)oxy)piperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Int-108e) (R)-(1-(((4-(3-methyl-3-((trimethylsilyl)oxy)piperidin-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanol (Int-108c) (40.5 mg, 0.0996 mmol), (R)-(1-(((4-(3-methyl-3-((trimethylsilyl)oxy)piperidin-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methyl To a solution of acetate (Int-108d) (105 mg, 0.234 mmol), 3-hydroxy-8-iodo-1-naphthoic acid (Int-T002) (115 mg, 0.366 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (69.3 mg, 0.361 mmol), and 1-hydroxybenzotriazole hydrate (57.0 mg, 0.372 mmol) in DMF (3.4 mL) was added N,N-diisopropylethylamine (0.174 mL, 0.999 mmol) at room temperature. The mixture was stirred at room temperature for 3 hours, and then MeOH (1.0 mL) and 2.0 M aqueous NaOH (1.0 mL) were added to the reaction mixture. The mixture was stirred at room temperature for 20 minutes, and then quenched by the addition of 2.0 M aqueous HCl (1.0 mL). The reaction mixture was extracted with EtOAc, washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (0% to 40%, MeOH gradient / EtOAc) to give (R)-(3-hydroxy-8-iodonaphthalen-1-yl)(2-((1-(hydroxymethyl)cyclopropyl)methoxy)-4-(3-methyl-3-((trimethylsilyl)oxy)piperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Int-108e). ESI-MS m / z [M+H] + 703.

[0239] Example 108: (R)-(4-(3-hydroxy-3-methylpiperidin-1-yl)-2-((1-((4-methoxypiperidin-1-yl)methyl)cyclopropyl)methoxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 108) [ka]

[0240] To a solution of (R)-(3-hydroxy-8-iodonaphthalen-1-yl)(2-((1-(hydroxymethyl)cyclopropyl)methoxy)-4-(3-methyl-3-((trimethylsilyl)oxy)piperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Int-108e) (22.0 mg, 0.0313 mmol) in N,N-dimethylacetamide (0.4 mL) was added ethanesulfonyl chloride (0.00890 mL, 0.0939 mmol) and triethylamine (0.0220 mL, 0.157 mmol) at 5° C. The mixture was stirred at 5° C. for 15 minutes, and then 4-methoxypiperidine (0.0194 mL, 0.157 mmol) was added to the reaction mixture. After stirring the mixture at 60° C. for 3 hours, 2.0 M aqueous NaOH (0.1 mL) was added to the mixture, and the mixture was stirred at 60° C. for 30 minutes. 6.0 M aqueous HCl (0.1 mL) was added to the reaction mixture, and the reaction was stirred at 60° C. for 10 minutes. The reaction mixture was cooled to room temperature, diluted with DMSO (0.5 mL), and purified by reverse-phase HPLC (MeCN / water with 0.1% formic acid) to give (R)-(4-(3-hydroxy-3-methylpiperidin-1-yl)-2-((1-((4-methoxypiperidin-1-yl)methyl)cyclopropyl)methoxy)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 108). ESI-MS m / z [M+H] + 728.

[0241] The compounds in the following table were synthesized using Int-108e and the corresponding amines via a similar route to Example 108.

[0242] Table 4 [Table 4] TIFF2024521979000072.tif255153

[0243] Example 112: (2-((1-(((R)-3-fluoropyrrolidin-1-yl)methyl)cyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 112) [ka]

[0244] Step A: tert-Butyl 2-((1-(((R)-3-fluoropyrrolidin-1-yl)methyl)cyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-112a) To a solution of tert-butyl (R)-2-chloro-4-(3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-95a) (100 mg, 0.271 mmol), (R)-(1-((3-fluoropyrrolidin-1-yl)methyl)cyclopropyl)methanol (93.9 mg, 0.542 mmol), and RuPhosPdG (6.80 mg, 0.00813 mmol) in 1,4-dioxane (1.5 mL) was added cesium carbonate (265 mg, 0.813 mmol) at room temperature. The mixture was stirred at 100° C. for 3 hours, after which the reaction mixture was cooled to room temperature, filtered, and concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (50% to 100%, EtOAc gradient / hexanes) to give tert-butyl 2-((1-(((R)-3-fluoropyrrolidin-1-yl)methyl)cyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate ( Int-112a ) (156 mg) was obtained. ESI-MS m / z [M+H] + 506.

[0245] Step B: Example 112 (2-((1-(((R)-3-fluoropyrrolidin-1-yl)methyl)cyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 112) tert-Butyl 2-((1-(((R)-3-fluoropyrrolidin-1-yl)methyl)cyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate ( Int-112a To a solution of 137 mg (0.271 mmol) of HCl (4.0 M in dioxane) in 3.0 mL of chloroform was added 0.68 mL of HCl. After stirring at room temperature for 4 days, the reaction mixture was concentrated under reduced pressure to give the amine product.

[0246] To a solution of the amine product, 3-hydroxy-8-iodo-1-naphthoic acid (Int-T002) (93.7 mg, 0.298 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (57.2 mg, 0.298 mmol), and 1-hydroxybenzotriazole hydrate (45.7 mg, 0.298 mmol) in DMF (3.0 mL) was added N,N-diisopropylethylamine (0.472 mL, 2.71 mmol) at room temperature. The mixture was stirred at room temperature for 24 h and then quenched by the addition of HO. The reaction mixture was extracted with EtOAc, washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase HPLC (MeCN / water + 0.1% formic acid) to give (2-((1-(((R)-3-fluoropyrrolidin-1-yl)methyl)cyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 112). ESI-MS m / z [M+H] + 702. 1 H NMR (400 MHz, DMSO-d6) δ 10.3 (s, 1H), 8.02-7.96 (m, 1H), 7.87-7.80 (m, 1H), 7.29-7.23 (m, 1H), 7.16-7.07 (m, 2H), 5.26-4.83 (m, 2H), 4.68-4.33 (m, 2H), 4.20-3.93 (m, 4H), 3.87-3.65 (m, 1H), 2.87-2.68 (m, 2H), 2.34-2.22 (m, 2H), 2.16-1.95 (m, 1H), 1.91-1.70 (m, 2H), 1.68-1.40 (m, 3H), 1.34-1.20 (m, 2H), 1.13 (s, 3H), 0.89-0.75 (m, 3H), 0.60-0.47 (m, 2H), 0.45-0.31 (m, 2H).

[0247] The compounds in the following table were synthesized using Int-95a and the corresponding amino alcohols via a route similar to that of Example 112.

[0248] Table 5 [Table 5]

[0249] Example 117: (2-(((R)-2,2-difluoro-1-(hydroxymethyl)cyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 117) [ka]

[0250] Step A: tert-Butyl 2-(((R)-1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-117a) To a solution of tert-butyl (R)-2-chloro-4-(3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-95a) (400 mg, 1.08 mmol), (S)-(1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methanol (Int-UU5-2) (371 mg, 1.63 mmol), and RuPhosPdG (27.2 mg, 0.0325 mmol) in 1,4-dioxane (5.0 mL) was added cesium carbonate (1.06 g, 3.25 mmol) at room temperature. The mixture was stirred at 100 °C for 6 hours, after which the reaction mixture was cooled to room temperature, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (10% to 100%, EtOAc gradient / hexanes) to give tert-butyl 2-(((R)-1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-117a) (466 mg). ESI-MS m / z [M+H] + 561.

[0251] Step B: (R)-1-(2-(((R)-2,2-difluoro-1-(hydroxymethyl)cyclopropyl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (Int-117b) To a solution of tert-butyl 2-(((R)-1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-117a) (466 mg, 0.830 mmol) in ethanol (5.0 mL) was added Pd(OH)2 / C (400 mg, 0.570 mmol). The reaction mixture was purged with H2 gas and evacuated three times and stirred at room temperature for 2 hours. The reaction mixture was filtered and concentrated under reduced pressure. To a solution of the resulting product in dichloromethane (8.0 mL) was added trifluoroacetic acid (1.6 mL). The mixture was stirred at room temperature for 2 hours, and then the reaction mixture was concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (0% to 50%, MeOH gradient / EtOAc) to give (R)-1-(2-(((R)-2,2-difluoro-1-(hydroxymethyl)cyclopropyl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (Int-117b) (259 mg). ESI-MS m / z [M+H] + 371.

[0252] Step C: Example 117 (2-(((R)-2,2-difluoro-1-(hydroxymethyl)cyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 117) To a solution of (R)-1-(2-(((R)-2,2-difluoro-1-(hydroxymethyl)cyclopropyl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (Int-117b) (259 mg, 0.699 mmol), 3-hydroxy-8-iodo-1-naphthoic acid (Int-T002) (241 mg, 0.769 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (147 mg, 0.769 mmol) and 1-hydroxybenzotriazole hydrate (118 mg, 0.769 mmol) in DMF (7.0 mL) was added N,N-diisopropylethylamine (0.609 mL, 3.49 mmol) at room temperature. The mixture was stirred at room temperature for 9 hours and then quenched by the addition of H2O. The reaction mixture was extracted with EtOAc, washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (0% to 50%, MeOH gradient / EtOAc) and reverse-phase HPLC (MeCN / water + 0.1% formic acid) to give (2-(((R)-2,2-difluoro-1-(hydroxymethyl)cyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 117). ESI-MS m / z [M+H] + 667. 1 H NMR (400 MHz, DMSO-d6) δ 10.3 (s, 1H), 8.02-7.96 (m, 1H), 7.87-7.80 (m, 1H), 7.29-7.24 (m, 1H), 7.17-7.07 (m, 2H), 5.10-4.84 (m, 2H), 4.71-4.48 (m, 2H), 4.48-4.30 (m, 2H), 4.27-4.11 (m, 2H), 4.08-3.67 (m, 1H), 3.62-3.49 (m, 2H), 3.10-2.98 (m, 1H), 1.86-1.24 (m, 7H), 1.14 (s, 3H), 0.86-0.76 (m, 1H).

[0253] Example 118: (2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 118) [ka]

[0254] To a solution of (2-(((R)-2,2-difluoro-1-(hydroxymethyl)cyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 117) (29.1 mg, 0.0437 mmol) in N,N-dimethylacetamide (0.45 mL) was added ethanesulfonyl chloride (0.00869 mL, 0.0917 mmol) and triethylamine (0.0614 mL, 0.437 mmol) at 5° C. After stirring the mixture at 5° C. for 15 minutes, a 2.0 M solution of dimethylamine in THF (0.220 mL, 0.440 mmol) was added to the reaction mixture. After stirring the mixture at 75° C. for 3 hours, 2.0 M aqueous NaOH (0.4 mL) was added to the mixture, and the mixture was stirred at 75° C. for 30 minutes. The reaction mixture was cooled to room temperature, diluted with DMSO (0.4 mL), and purified by reverse-phase HPLC (MeCN / water with 0.1% formic acid) to give (2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 118). ESI-MS m / z [M+H] + 694. 1H NMR (400 MHz, DMSO-d6) δ 10.3 (s, 1H), 8.02-7.95 (m, 1H), 7.87-7.80 (m, 1H), 7.29-7.23 (m, 1H), 7.16-7.07 (m, 2H), 5.07-4.86 (m, 1H), 4.72-3.99 (m, 6H), 3.91-3.67 (m, 1H), 3.60-3.47 (m, 1H), 3.13-2.98 (m, 1H), 2.25 (d, J = 12.7 Hz, 1H), 2.19-2.07 (m, 6H), 1.85-1.35 (m, 6H), 1.14 (s, 3H), 0.94-0.75 (m, 2H).

[0255] Using Example 117 and the corresponding amine, the compounds in the following table were synthesized by a similar route to Example 118.

[0256] Table 6 [Table 6] TIFF2024521979000078.tif25535

[0257] Example 124: (2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 124) [ka]

[0258] Step A: tert-Butyl 2-(((R)-1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-124a) To a solution of tert-butyl (R)-2-chloro-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-84a-3) (200 mg, 0.545 mmol), (S)-(1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methanol (Int-UU5-2) (187 mg, 0.818 mmol), and RuPhosPdG (13.7 mg, 0.0164 mmol) in 1,4-dioxane (1.5 mL) was added cesium carbonate (533 mg, 1.64 mmol) at room temperature. The mixture was stirred at 100 °C for 5 h, after which the reaction mixture was cooled to room temperature, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 50%, EtOAc gradient / hexanes) to give tert-butyl 2-(((R)-1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-124a) (305 mg). ESI-MS m / z [M+H] + 559.

[0259] Step B: tert-Butyl 2-(((R)-2,2-difluoro-1-(hydroxymethyl)cyclopropyl)methoxy)-4-((R)-2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-124b) To a solution of tert-butyl 2-(((R)-1-((benzyloxy)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-124a) (305 mg, 0.545 mmol) in ethanol (5.5 mL) was added Pd(OH)2 / C (191 mg, 0.273 mmol). The reaction mixture was purged with H2 gas and evacuated three times and stirred at room temperature for 2 hours. The reaction mixture was filtered and concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (30% to 100%, EtOAc gradient / hexanes) to give tert-butyl 2-(((R)-2,2-difluoro-1-(hydroxymethyl)cyclopropyl)methoxy)-4-((R)-2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-124b) (253 mg). ESI-MS m / z [M+H] + 469.

[0260] Step C: tert-Butyl 2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-124c) To a solution of tert-butyl 2-(((R)-2,2-difluoro-1-(hydroxymethyl)cyclopropyl)methoxy)-4-((R)-2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-124b) (253 mg, 0.539 mmol) in N,N-dimethylacetamide (5.0 mL) was added ethanesulfonyl chloride (0.0562 mL, 0.593 mmol) and triethylamine (0.379 mL, 2.70 mmol) at 5 °C. The mixture was stirred at 5 °C for 10 minutes, and then a 2.0 M solution of dimethylamine in THF (1.35 mL, 2.70 mmol) was added to the reaction mixture. The mixture was stirred at 60 °C for 14 hours and then quenched by the addition of HO. The reaction mixture was extracted with EtOAc, washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (30% to 100%, EtOAc gradient / hexanes) to give tert-butyl 2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-124c) (216 mg). ESI-MS m / z [M+H] + 496.

[0261] Step D: Example 124 (2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 124) To a solution of tert-butyl 2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-124c) (216 mg, 0.435 mmol) in chloroform (4.0 mL) was added 4.0 M HCl in dioxane (2.2 mL). After stirring the mixture at room temperature for 30 minutes, the reaction mixture was concentrated under reduced pressure to provide the amine product.

[0262] To a solution of the amine product, 3-hydroxy-8-iodo-1-naphthoic acid (Int-T002) (97.0 mg, 0.309 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (59.2 mg, 0.309 mmol), and 1-hydroxybenzotriazole hydrate (47.3 mg, 0.309 mmol) in DMF (3.0 mL) at room temperature was added N,N-diisopropylethylamine (0.244 mL, 1.40 mmol). The mixture was stirred at room temperature for 13 h and then quenched by the addition of HO. The reaction mixture was extracted with EtOAc, washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase HPLC (MeCN / water with 0.1% formic acid) to give (2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 124). ESI-MS m / z [M+H] + 692. 1 H NMR (400 MHz, DMSO-d6) δ 10.2 (s, 1H), 8.03-7.96 (m, 1H), 7.87-7.80 (m, 1H), 7.29-7.23 (m, 1H), 7.19-7.07 (m, 2H), 5.16-4.85 (m, 1H), 4.79-4.00 (m, 5H), 3.80-3.55 (m, 1H), 3.05-2.85 (m, 1H), 2.37-2.20 (m, 2H), 2.17-2.06 (m, 6H), 1.96-1.56 (m, 4H), 1.55-1.25 (m, 4H), 1.25-0.52 (m, 6H).

[0263] Example 125-1: (3-amino-8-iodonaphthalen-1-yl)(2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 125) [ka]

[0264] Step A: tert-Butyl 2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-125a) To a solution of tert-butyl (R)-2-chloro-4-(3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-95a) (100 mg, 0.271 mmol), (R)-(1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methanol (Int-UU8-1) hydrochloride (82.0 mg, 0.407 mmol) and RuPhosPdG3 (6.80 mg, 0.00813 mmol) in 1,4-dioxane (1.5 mL) was added cesium carbonate (265 mg, 0.813 mmol) and N,N-diisopropylethylamine (0.0944 mL, 0.542 mmol) at room temperature. After stirring the mixture at 100° C. for 14 hours, the reaction mixture was cooled to room temperature, filtered, and concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (20% to 100%, EtOAc gradient / hexanes) to give tert-butyl 2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-125a) (104 mg). ESI-MS m / z [M+H] + 498.

[0265] Step B: (R)-1-(2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (Int-125b) To a solution of tert-butyl 2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidine-6-carboxylate (Int-125A) (104 mg, 0.210 mmol) in dichloromethane (2.0 mL) was added trifluoroacetic acid (0.4 mL). The mixture was stirred at room temperature for 2 hours, and then the reaction mixture was concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (0% to 50%, MeOH gradient / EtOAc) to give (R)-1-(2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (Int-125b) (72.8 mg). ESI-MS m / z [M+H] + 398.

[0266] Step C: Example 125-1 (3-amino-8-iodonaphthalen-1-yl)(2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 125-1) (R)-1-(2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol (Int-125b) (6.00 mg, 0.0151 mmol), 3-amino-8-iodo-naphthalene-1-carboxylic acid (Int-T001) (5 To a solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.18 mg, 0.0166 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.18 mg, 0.0166 mmol), and 1-hydroxybenzotriazole hydrate (2.54 mg, 0.0166 mmol) in DMF (0.15 mL) was added N,N-diisopropylethylamine (0.0131 mL, 0.0755 mmol) at room temperature. The mixture was stirred at room temperature for 14 h and then quenched by the addition of HO. The reaction mixture was extracted with EtOAc, washed with brine, dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by reverse-phase HPLC (MeCN / water + 0.1% formic acid) to give (3-amino-8-iodonaphthalen-1-yl)(2-(((R)-1-((dimethylamino)methyl)-2,2-difluorocyclopropyl)methoxy)-4-((R)-3-hydroxy-3-methylpiperidin-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 125-1). ESI-MS m / z [M+H] + 693. 1 H NMR (400 MHz, DMSO-d6) δ 7.82-7.77 (m, 1H), 7.67-7.60 (m, 1H), 7.03-6.96 (m, 2H), 6.94-6.89 (m, 1H), 5.69 (s, 2H), 5.06-4.80 (m, 1H), 4.65-4.00 (m, 6H), 3.57-3.47 (m, 2H), 2.18-2.06 (m, 6H), 1.84-1.35 (m, 6H), 1.25-1.19 (m, 2H), 1.14 (s, 3H), 0.89-0.76 (m, 2H).

[0267] Example 125-2: (3-amino-8-iodonaphthalen-1-yl)(2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 125-2) [ka]

[0268] To a mixture of 3-amino-8-iodo-1-naphthoic acid (Int-T001) (52 mg, 0.167 mmol), 1-hydroxybenzotriazole monohydrate (26 mg, 0.167 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (32 mg, 0.167 mmol), and N,N-diisopropylethylamine (0.071 mL, 0.417 mmol) in N,N-dimethylformamide (2 mL) was added N,N-dimethyl-1-(1-(((4-(2-methylazepan-1-yl)-6,7-dihydro-5H-pyrrolo[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanamine (Int-84b) (50 mg, 0.139 mmol), and the mixture was stirred at room temperature for 3 hours. The mixture was diluted with ethyl acetate, the diluted mixture was washed with water, and the mixture was concentrated under reduced pressure. The residue was purified by reverse phase HPLC (MeCN / water + 0.1% formic acid) to give (3-amino-8-iodonaphthalen-1-yl)(2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)methanone (Example 125-2) (30 mg). ESI-MS m / z [M+H] + 655.

[0269] Example 126: (2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(8-ethynyl-3-hydroxynaphthalen-1-yl)methanone (Example 126) [ka]

[0270] To a solution of copper(I) iodide (2.9 mg, 0.0153 mmol), bis(triphenylphosphine)palladium(II) dichloride (11 mg, 0.0153 mmol), (2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(3-hydroxy-8-iodonaphthalen-1-yl)methanone (Example 84) (100 mg, 0.0153 mmol), and triethylamine (0.107 mL, 0.763 mmol) in N,N-dimethylformamide (3 mL) was added triisopropylsilylacetylene (0.102 mL, 0.458 mmol). The vessel was evacuated and backfilled with nitrogen, and the mixture was stirred at 95° C. for 0.5 hours. The mixture was diluted with ethyl acetate, the diluted mixture was washed with water, and the mixture was concentrated under reduced pressure. The residue was purified by flash NH-silica gel chromatography (0% to 40%, methanol gradient / ethyl acetate) to give the coupling product.

[0271] To a solution of the coupling product in tetrahydrofuran (2 mL) was added tetrabutylammonium fluoride (0.229 mL, 0.229 mmol, 1 M solution in tetrahydrofuran), and the mixture was stirred at room temperature for 0.5 hours. The mixture was diluted with ethyl acetate, the diluted mixture was washed with water, and the mixture was concentrated under reduced pressure. The residue was purified by reverse-phase HPLC (MeCN / water with 0.1% formic acid) to give (2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-(2-methylazepan-1-yl)-5,7-dihydro-6H-pyrrolo[3,4-d]pyrimidin-6-yl)(8-ethynyl-3-hydroxynaphthalen-1-yl)methanone (Example 126). ESI-MS m / z [M+H] + 554. 1H NMR (400 MHz, DMSO-d6) δ 10.2 (s, 1H), 7.89-7.84 (m, 1H), 7.58-7.52 (m, 1H), 7.48-7.39 (m, 1H), 7.31-7.26 (m, 1H), 7.12-7.06 (m, 1H), 5.20-4.80 (m, 2H), 4.61-4.30 (m, 2H), 4.17-3.95 (m, 4H), 3.13-2.75 (m, 1H), 2.25-2.08 (m, 2H), 2.16 (s, 3H), 2.11 (s, 3H), 2.00-0.95 (m, 12H), 0.59-0.48 (m, 2H), 0.39-0.31 (m, 2H).

[0272] The compounds in the following table were synthesized by a similar route to Example 126 using the corresponding iodide samples.

[0273] Table 7 [Table 7] TIFF2024521979000084.tif153170TIFF2024521979000085.tif133170TIFF20245219790 00086.tif132170TIFF2024521979000087.tif152170TIFF2024521979000088.tif132170

[0274] 7-(8-Bromo-3-(methoxymethoxy)naphthalen-1-yl)-2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-ol (Int-144e-1) and Int-144e-2 isomers (mixture) [ka]

[0275] Step A: tert-Butyl 2-chloro-4-((2-nitrobenzyl)oxy)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (Int-144a) Cesium carbonate (42.9 g, 0.131 mmol) was added to a stirred solution of tert-butyl 2,4-dichloro-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (20.0 g, 65.7 mmol) and (2-nitrophenyl)methanol (13.1 g, 85.5 mmol) in toluene (200 mL), and the mixture was warmed to 80°C and stirred for 3 hours. The mixture was cooled to room temperature and diluted with ethyl acetate (200 mL). The mixture was washed twice with water (200 mL), and the organic phase was concentrated under reduced pressure. MeCN (300 mL) and water (60 mL) were added to the residue, and the mixture was suspended at 80°C for 1 hour. The suspension was cooled to room temperature, and the solid was filtered and dried under vacuum to give tert-butyl 2-chloro-4-((2-nitrobenzyl)oxy)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (Int-144a) (15.7 g). ESI-MS m / z [M+H] + 421, 423.

[0276] Step B: tert-Butyl 2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-((2-nitrobenzyl)oxy)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (Int-144b) tert-Butyl 2-chloro-4-((2-nitrobenzyl)oxy)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate ( Int-144aTo a mixture of (1.96 g, 4.66 mmol), (1-((dimethylamino)methyl)cyclopropyl)methanol (1.20 g, 9.29 mmol), and dichloro[9,9-dimethyl-4,5-bis(diphenylphosphino)xanthene]palladium(II) (352 mg, 0.466 mmol) in toluene (20 mL) was added cesium carbonate (4.55 g, 14.0 mmol), and the mixture was warmed to 110 °C and stirred for 3 h. The mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 25%, MeOH gradient / chloroform). The resulting residue was purified by flash NH-silica gel chromatography (0% to 80%, ethyl acetate gradient / hexane) to give tert-butyl 2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-((2-nitrobenzyl)oxy)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate ( Int-144b ) (1.90 g) was obtained. ESI-MS m / z [M+H] + 514.

[0277] Step C: N,N-Dimethyl-1-(1-(((4-((2-nitrobenzyl)oxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanamine (Int-144c) tert-Butyl 2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-4-((2-nitrobenzyl)oxy)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate ( Int-144b To a solution of N,N-dimethyl-1-(1-(((4-((2-nitrobenzyl)oxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanamine ( Int-144c ) (1.64 g) was obtained. ESI-MS m / z [M+H] + 414.

[0278] Step D: 1-(1-(((7-(8-bromo-3-(methoxymethoxy)naphthalen-1-yl)-4-((2-nitrobenzyl)oxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)-N,N-dimethylmethanamine (Int-144d-1) and Int-144d-2 isomers (mixture) N,N-Dimethyl-1-(1-(((4-((2-nitrobenzyl)oxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)methanamine (Int-144c) (1.65 g, 3.99 mmol), 1,8-dibromo-3-(methoxymethoxy)naphthalene (Int-W002) (2.76 g, 7.98 mmol), dichloro[9,9-dimethyl-4,5-bis(diphenylphosphino)xanthene]palladium(II) (905 mg, 1.20 mmol), and cesium carbonate (3.90 g, 12.0 mmol) in toluene (16 mL) were placed in a sealed tube. The reaction vessel was evacuated and backfilled with nitrogen three times. The mixture was stirred at 125 °C for 14 h, and the mixture was then cooled to room temperature. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 25%, MeOH gradient / chloroform). The resulting residue was purified by flash NH-silica gel chromatography (0% to 8%, ethyl acetate gradient / hexane) to give 1-(1-(((7-(8-bromo-3-(methoxymethoxy)naphthalen-1-yl)-4-((2-nitrobenzyl)oxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)-N,N-dimethylmethanamine (Int-144d-1) (400 mg) as a mixture of Int-144d-2 isomers. ESI-MS m / z [M+H] + 678, 680.

[0279] Step E: 7-(8-Bromo-3-(methoxymethoxy)naphthalen-1-yl)-2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-ol (Int-144e-1) and Isomer (Mixture) (Int-144e-2) To a mixture of 1-(1-(((7-(8-bromo-3-(methoxymethoxy)naphthalen-1-yl)-4-((2-nitrobenzyl)oxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2-yl)oxy)methyl)cyclopropyl)-N,N-dimethylmethanamine (Int-144d-1) and Int-144d-2 isomers (255 mg, 0.376 mmol) and iron (powder, 210 mg, 3.76 mmol) in tetrahydrofuran (6.0 mL) was added hydrochloric acid (0.5 M, 6.0 mL), and the mixture was stirred at room temperature for 2 hours. The mixture was filtered, and the filtrate was concentrated to dryness under reduced pressure. The residue was purified by flash NH-silica gel chromatography (0% to 100%, MeOH gradient / ethyl acetate) to give 7-(8-bromo-3-(methoxymethoxy)naphthalen-1-yl)-2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-ol (Int-144e-1) as a mixture of Int-144e-2 isomers (180 mg). ESI-MS m / z [M+H] + 543, 545.

[0280] 7-(8-Bromo-3-(methoxymethoxy)naphthalen-1-yl)-2-((1-(morpholinomethyl)cyclopropyl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-ol (Int-144f-1) and Int-144f-2 isomers (mixture) [ka]

[0281] A mixture of isomers, Int-144f-1 and Int-144f-2, was synthesized via a similar route to 7-(8-bromo-3-(methoxymethoxy)naphthalen-1-yl)-2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-ol (Int-144e-1) using (1-(morpholinomethyl)cyclopropyl)methanol. ESI-MS m / z [M+H] + 585, 587.

[0282] Example 144: (R)-1-(7-(8-bromo-3-hydroxynaphthalen-1-yl)-2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperidin-3-ol (Example 144) [ka]

[0283] To a solution of 7-(8-bromo-3-(methoxymethoxy)naphthalen-1-yl)-2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-ol (Int-144e-1) (33 mg, 0.0607 mmol) and N,N-diisopropylethylamine (0.031 mL, 0.182 mmol) in dichloromethane (1 mL) was added p-toluenesulfonyl chloride (18 mg, 0.0911 mmol), and the mixture was stirred at room temperature for 3 hours. (R)-Piperidin-3-ol (20 mg, 0.198 mmol) was added to the mixture, and the mixture was stirred at 60° C. for 3 hours. The solvent was removed under reduced pressure, and to the residue was added hydrochloride (4 M in 1,4-dioxane, 2 mL), and the mixture was evaporated to dryness. The residue was purified by reverse-phase HPLC (MeCN / water with 0.1% formic acid) to give (R)-1-(7-(8-bromo-3-hydroxynaphthalen-1-yl)-2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperidin-3-ol (Example 144). ESI-MS m / z [M+H] + 582, 584. 1H NMR (400 MHz, DMSO-d6) δ 9.94 (s, 1H), 7.73-7.68 (m, 1H), 7.56-7.25 (m, 1H), 7.23-7.18 (m, 1H), 6.98-6.95 (m, 1H), 6.91-6.87 (m, 1H), 4.65 (s, 0.5H), 4.42 (s, 0.5H), 4.15-4.00 (m, 4H), 3.62-3.05 (m, 4H), 2.63-2.52 (m, 2H), 2.25-2.19 (m, 2H), 2.16 (s, 6H), 1.95-1.40 (m, 5H), 1.28-1.22 (m, 1H), 1.15 (s, 1.5H), 1.07 (s, 1.5H), 0.58-0.55 (m, 2H), 0.38-0.35 (m, 2H).

[0284] The compounds in the following table were synthesized using 7-(8-bromo-3-(methoxymethoxy)naphthalen-1-yl)-2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-ol (Int-144e-1) or 7-(8-bromo-3-(methoxymethoxy)naphthalen-1-yl)-2-((1-(morpholinomethyl)cyclopropyl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-ol (Int-144f-1) and the corresponding amine via a route similar to that described in Example 144.

[0285] Table 8 [Table 8] TIFF2024521979000093.tif164118

[0286] (3R)-3-Methyl-1-(7-(6-methyl-1-(tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)-1H-indazol-4-yl)-2-(methylthio)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperidin-3-ol (Int-149e) [ka]

[0287] Step A: tert-Butyl 4-hydroxy-2-(methylthio)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (Int-149a) To a solution of 1-(tert-butyl) 4-ethyl 3-oxopiperidine-1,4-dicarboxylate (10.0 g, 36.9 mmol) and S-methylisothiourea sulfate (20.5 g, 73.7 mmol) in methanol (200 mL) was added sodium methoxide (9.96 g, 184 mmol), and the mixture was stirred at 60 °C for 4 hours. The reaction mixture was cooled to room temperature, and hydrochloric acid (6 M, 20.0 mL, 120 mmol) was added to the mixture. Methanol was removed under reduced pressure, and water (150 mL) and ethyl acetate (150 mL) were added to the residue. The precipitate was filtered, washed with ethyl acetate (30 mL), and dried to give tert-butyl 4-hydroxy-2-(methylthio)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (Int-149a) (4.70 g). ESI-MS m / z [M+H] + 298.

[0288] Step B: tert-Butyl 2-(methylthio)-4-(((trifluoromethyl)sulfonyl)oxy)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (Int-149b) To a solution of tert-butyl 4-hydroxy-2-(methylthio)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (Int-149a) (2.00 g, 6.73 mmol) and N,N-diisopropylethylamine (3.43 mL, 20.2 mmol) in dichloromethane (20 mL) was added trifluoromethanesulfonic anhydride (1.10 mL, 6.73 mmol) at 0° C., and the mixture was stirred at room temperature for 1 hour. The mixture was diluted with ethyl acetate, washed with water, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 30%, ethyl acetate gradient / hexanes) to give tert-butyl 2-(methylthio)-4-(((trifluoromethyl)sulfonyl)oxy)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (Int-149b) (1.50 g). ESI-MS m / z [M+H] + 430.

[0289] Step C: tert-Butyl (R)-4-(3-hydroxy-3-methylpiperidin-1-yl)-2-(methylthio)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (Int-149c) To a solution of tert-butyl 2-(methylthio)-4-(((trifluoromethyl)sulfonyl)oxy)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (Int-149b) (600 mg, 1.40 mmol) and (R)-3-methylpiperidin-3-ol hydrochloride (233 mg, 1.54 mmol) in DMA (10 mL) was added N,N-diisopropylethylamine (3.43 mL, 20.2 mmol), and the mixture was stirred at room temperature for 3 hours. The mixture was diluted with ethyl acetate, the diluted mixture was washed with water, and the mixture was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0% to 80%, ethyl acetate gradient / hexanes) to give tert-butyl (R)-4-(3-hydroxy-3-methylpiperidin-1-yl)-2-(methylthio)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (Int-149c) (570 mg). ESI-MS m / z [M+H]+ 395.

[0290] Step D: (R)-3-Methyl-1-(2-(methylthio)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperidin-3-ol (Int-149d) To a solution of tert-butyl (R)-4-(3-hydroxy-3-methylpiperidin-1-yl)-2-(methylthio)-5,8-dihydropyrido[3,4-d]pyrimidine-7(6H)-carboxylate (Int-149c) (570 mg, 1.44 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (5 mL), and the mixture was stirred at room temperature for 1 hour. The trifluoroacetic acid was removed under reduced pressure, and the residue was purified by flash NH-silica gel chromatography (0% to 20%, MeOH gradient / ethyl acetate) to give (R)-3-methyl-1-(2-(methylthio)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperidin-3-ol (Int-149d) (430 mg). ESI-MS m / z [M+H]+ 295.

[0291] Step E: (3R)-3-Methyl-1-(7-(6-methyl-1-(tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)-1H-indazol-4-yl)-2-(methylthio)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperidin-3-ol (Int-149e) To a mixture of (R)-3-methyl-1-(2-(methylthio)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperidin-3-ol (Int-149d) (150 mg, 0.509 mmol), 4-bromo-6-methyl-1-(tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)-1H-indazole (Int-HHH5) (278 mg, 0.764 mmol), and RuPhosPdG3 (42.6 mg, 0.0509 mmol) in 1,4-dioxane (2.0 mL) was added cesium carbonate (498 mg, 1.53 mmol), and the mixture was warmed to 100 °C and stirred for 2 h. The mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (10% to 70%, ethyl acetate gradient / hexanes) to give (3R)-3-methyl-1-(7-(6-methyl-1-(tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)-1H-indazol-4-yl)-2-(methylthio)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperidin-3-ol (Int-149e) (175 mg). ESI-MS m / z [M+H] + 577.

[0292] Example 149: (R)-1-(2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-7-(6-methyl-5-(trifluoromethyl)-1H-indazol-4-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol formate (Example 149) [ka]

[0293] To a solution of (3R)-3-methyl-1-(7-(6-methyl-1-(tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)-1H-indazol-4-yl)-2-(methylthio)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)piperidin-3-ol (Int-149e) (15 mg, 0.0260 mmol) in ethyl acetate (3 mL) was added 3-chloroperbenzoic acid (6.4 mg, 0.0260 mmol) at 0° C., and the mixture was stirred at 0° C. for 0.5 hours. Water (3 mL) was added to the mixture, and the mixture was washed with saturated sodium bicarbonate solution (3 mL). The solvent was removed under reduced pressure. To a solution of the crude product and (1-((dimethylamino)methyl)cyclopropyl)methanol (10 mg, 0.078 mmol) in tetrahydrofuran (1 mL) was added potassium tert-butoxide (1.0 M solution in THF, 0.039 mL) at 0° C., and the mixture was stirred at 0° C. for 0.5 hours. The mixture was diluted with ethyl acetate, and the diluted mixture was washed with water, and the mixture was concentrated under reduced pressure. Trifluoroacetic acid (0.5 mL) was added to the residue, and the mixture was stirred at room temperature for 1 hour. The trifluoroacetic acid was removed under reduced pressure, and the residue was purified by reverse-phase HPLC (MeCN / water with 0.1% formic acid) to give (R)-1-(2-((1-((dimethylamino)methyl)cyclopropyl)methoxy)-7-(6-methyl-5-(trifluoromethyl)-1H-indazol-4-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-3-methylpiperidin-3-ol formate (Example 149). ESI-MS m / z [M+H] + 574. 1 H NMR (400 MHz, DMSO-d6) δ 13.4 (s, 1H), 8.32 (s, 1H), 7.36 (s, 1H), 4.60-4.44 (m, 1H), 4.24-4.04 (m, 4H), 3.62-3.20 (m, 9H), 2.90-2.60 (m, 2H), 2.58-2.54 (m, 3H), 2.38-2.10 (m, 4H), 1.90-1.75 (m, 1H), 1.62-1.53 ​​(m, 4H), 1.12 (s, 3H), 0.79-0.37 (m, 4H).

[0294] The compounds in the following table were synthesized using the corresponding amino alcohols via a similar route to Example 149.

[0295] Table 9 [Table 9]

[0296] Assay SOS-Catalyzed Nucleotide Exchange Assay Procedure: Recombinant KRAS G12C (amino acids 1-169, SEQ ID NO: 1), KRAS G12D (amino acids 1-169, SEQ ID NO: 2), KRAS G12V (amino acids 1-169, SEQ ID NO: 3), and SOS1 (amino acids 564-1049, SEQ ID NO: 4) proteins were expressed in Escherichia coli (E. coli) and purified by affinity chromatography. To prepare each BODIPY FL GDP-bound KRAS mutant protein, 50 μM KRAS mutant protein was incubated with 0.5 mM BODIPY FL GDP (Invitrogen, G22360) in loading buffer (20 mM Tris-HCl (pH 7.5), 50 mM NaCl, 1 mM DTT, and 2.5 mM EDTA) on ice for 1 h. After incubation, MgCl2 was added to a final concentration of 10 mM, and the mixture was incubated at room temperature for 30 min. The mixture was passed through a NAP-5 column to remove free nucleotides, and the purified BODIPY FL GDP-bound KRAS G12C, G12D, and G12V proteins were used for compound evaluation.

[0297] To measure the inhibitory activity of compounds on the GDP-GTP exchange rate of recombinant KRAS mutants, each BODIPY FL GDP-bound KRAS mutant protein (version 1: 25 nM, version 2: 2.5 nM) was incubated with various concentrations of compounds in reaction buffer (20 mM Tris-HCl (pH 7.5), 100 mM NaCl, 1 mM MgCl2, 2 mM DTT, 0.1% Tween 20) at 25°C for 1 h. After incubation, recombinant SOS1 and GMPPNP (Jena Bioscience GmbH, NU-401) were added and incubated at room temperature for 30 min to allow the SOS1-dependent GDP-GTP exchange reaction in the KRAS mutants to proceed. The displacement of BODIPY FL GDP by guanosine-5'-[(β,γ)-imido]triphosphate tetralithium salt (GMPPNP) was measured by calculating the ratio of the fluorescence intensity of BODIPY FL before and after the exchange reaction. Percent inhibition was calculated by setting the fluorescence ratios from reactions without test compound (DMSO control) and reactions without SOS1 and GMPPNP as 0% and 100% inhibition, respectively. Dose-response curves were analyzed using a four-parameter logistic model to determine IC 50 The values ​​were calculated and the results are shown in Table A.

[0298] Table A: [Table 10] TIFF2024521979000098.tif255167TIFF2024521979000099.tif254167TIFF20245219790 00100.tif255167TIFF2024521979000101.tif254166TIFF2024521979000102.tif150168

[0299] Cytostatic assay procedure SW620 cells (ATCC®, CCL-227™) containing a homozygous KRAS-G12V activating mutation and MIA PaCa-2 cells (provided by Sumitomo Dainippon Pharma Co., Ltd.) containing a homozygous KRAS-G12C activating mutation were cultured in T175 flasks in growth medium (RPMI medium 1640 (Fujifilm Wako Pure Chemical Corporation, 187-02705) containing 10% fetal bovine serum (HyClone, SH30910.03)). After trypsin / EDTA (Nacalai Tesque, 32777-44) digestion, the cells were harvested in growth medium and seeded at a density of 250 cells / well into Prime Surface® 384-well U-bottom microplates (Sumitomo Bakelite Co., Ltd., MS-9384W) and incubated overnight at 37°C and 5% CO.

[0300] The test compound was diluted with DMSO to a concentration 500 times higher than the final concentration. The resulting DMSO solution of the test compound was diluted with the growth medium used to suspend the cells and added to each well of the cell culture plate to a final DMSO concentration of 0.2%, followed by incubation at 37°C, 5% CO2 for 3 days.

[0301] CellTiter-Glo® 3D reagent (Promega, G9683) was added to all cells and mixed for 10 minutes. Thirty minutes after mixing, luminescence was measured on an EnVision plate reader (PerkinElmer). Dose-response curves were analyzed using a four-parameter logistic model to determine the IC 50 The values ​​were calculated and the results are shown in Table B.

[0302] Table B: [Table 11] TIFF2024521979000104.tif255134TIFF2024521979000105.tif255133TIFF2024521979000106.tif254134TIFF2024521979000107.tif114133

[0303] array SEQ ID NO:1 - Recombinant KRASG12C

number

number

number

number

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof. 【Chemical 1】 [Wherein, X is (i)6- to 9-membered monocyclic or fused bicyclic or bridged bicyclic heterocyclic alkyl [said heterocyclic alkyl is saturated and contains 1 to 2 heteroatoms selected from the group consisting of N, S and O.]; (ii)8- to 10-membered spiro heterocyclic alkyl [said spiro heterocyclic alkyl is saturated and contains 1 to 2 heteroatoms selected from the group consisting of N and O.]; (iii) [Chemical Formula 2] and [Chemical Formula 3] selected from the group consisting of; When X is (i) or (ii), X is unsubstituted or is halo, hydroxy, C １ -C ６ alkyl, C １ -C ３ hydroxyalkyl, C １ -C ６ fluoroalkyl, carboxy, carbamoyl, C １ -C ３ carboxyalkyl, oxo, cyano, cyanomethyl, amino, pyrazolyl, oxadiazolonyl, -NHC(O)C １ -C ３ alkoxy C １ -C ３ alkyl, -NHC(O)C １ -C ３ alkoxy C ６ -C １０ aryl, C １ -C ３ alkoxy, methoxy (C １ -C ３ ), alkyl, amino (C １ -C ３ ), alkyl, C １ -C ３ alkylamino (C １ -C ３ ), alkyl, C １ -C ３ dialkylamino, C １ -C ３ dialkylamino (C １ -C ３ ), alkyl, and NHC(O)C ５ -C １０ heteroaryl, and is independently substituted by 1 to 4 R Ｘ substituents selected from the group consisting of; heteroaryl may be substituted by C １ -C ３ alkyl; Ring Y is a 9- to 10-membered bicyclic ring system, said ring system is partially unsaturated or aromatic, and ring Y contains 0 to 2 nitrogen heteroatoms; Ring Y is unsubstituted or is substituted by 1 to 4 R substituents independently selected from the group consisting of halo, hydroxy, amino, C １ -C ３ alkyl, C ２ -C ３ alkynyl, and C １ -C ３ fluoroalkyl; ｙ and is independently substituted by substituents; Z is (i) a 5- to 8-membered monocyclic or bicyclic heterocyclic alkyl [the heterocyclic alkyl is saturated, contains 1 nitrogen heteroatom, and the heterocyclic alkyl is unsubstituted or substituted with one substituent R selected from the group consisting of halo, C １ -C ３ alkyl, and methylene (C １ -C ３ alkyl)(C １ -C ３ alkyl)carbamate.]; ＺＨＣ is substituted with.]]; (ii) 【Chemical 4】 [wherein, M is selected from the group consisting of hydroxy, C １ -C ３ dialkylamino and C １ -C ４ alkylamino, and the cyclopropyl group is unsubstituted or substituted with a maximum of two halo groups.]; (iii) 【Chemical Formula 5】 [wherein, P is a 5- to 8-membered monocyclic, fused bicyclic or bridged bicyclic heterocyclic alkyl, the heterocyclic alkyl is saturated, contains 1 to 2 heteroatoms selected from the group consisting of N and O, and the heterocyclic alkyl is unsubstituted or substituted with halo, hydroxy, C １ -C ３ hydroxyalkyl, C １ -C ３ cyanoalkyl, carbamoyl, C １ -C ３ alkoxy, cyano, -NHC(O)C １ -C ３ alkyl, and one R Ｐ substituent selected from the group consisting of oxadiazolonyl; the cyclopropyl group is unsubstituted or substituted with a maximum of 2 halo groups.], and is selected from the group consisting of; Subscript m is 0 or 1; Subscript n is 1 or 2. ]

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein ring Y is 【Chemical Formula 6】

3. The compound according to claim 2, or a pharmaceutically acceptable salt thereof, wherein ring Y is 【Chemical Formula 7】

4. X is [Chemical Formula 8] X is unsubstituted or is substituted by 1 to 4 Rs Ｘ The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein X is unsubstituted or is independently substituted by a substituent

5. X is halo, hydroxy, C １ -C ６ alkyl, C １ -C ３ hydroxyalkyl, C １ -C ６ fluoroalkyl, carboxy, carbamoyl, C １ -C ３ carboxyalkyl, oxo, cyano, cyanomethyl, amino, pyrazolyl, oxadiazolonyl, -NHC(O)C １ -C ３ alkoxy C １ -C ３ alkyl, -NHC(O)C １ -C ３ alkoxy C ６ -C １０ aryl, and NHC(O)C ５ -C １０ selected from the group consisting of 1 to 4 R Ｘ substituents, wherein heteroaryl may be substituted by C １ -C ３ alkyl, the compound according to claim 4 or a pharmaceutically acceptable salt of the compound.

6. X is 【Chemical Formula 9】 【Chem.】 【Chem.】 【Chem.】

7. X is 【Chemical Formula 10】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein subscript p is 0, 1 or 2.

8. X is 【Chemical Formula 11】

9. Z is 【Chemical 12】 【Chem.】 [Chemical]

10. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein subscript m is 1.

11. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein subscript n is 1.

12. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein subscript n is 2.

13. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, selected from Examples 1 to 151.

14. A pharmaceutical composition comprising the compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

15. A pharmaceutical composition comprising the compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, another anti-cancer agent, and a pharmaceutically acceptable carrier.

16. A pharmaceutical composition for inhibiting KRAS-G12D protein, comprising contacting the KRAS-G12D protein with a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt of the compound to inhibit the activity of the KRAS-G12D protein.

17. A pharmaceutical composition for inhibiting KRAS-G12C protein, comprising contacting the KRAS-G12C protein with a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt of the compound to inhibit the activity of the KRAS-G12C protein.

18. A pharmaceutical composition for inhibiting KRAS-G12V protein, comprising contacting the KRAS-G12V protein with a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt of the compound to inhibit the activity of the KRAS-G12V protein.

19. A pharmaceutical composition for the treatment of cancer, comprising a therapeutically effective amount of a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt of the compound.

20. The pharmaceutical composition according to claim 19, comprising an additional active agent.

21. A compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt of the compound for use in a method of treatment.

22. A compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt of the compound for use in the treatment of cancer.

23. A compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt of the compound for the preparation of a medicament for the treatment of cancer, or the use of a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt of the compound for the preparation of a medicament for the treatment of cancer.

24. A compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt of the compound and an additional anti-cancer agent for use in the treatment of cancer.

25. A compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt of the compound and an additional anti-cancer agent for the preparation of a medicament for the treatment of cancer, or the use of a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt of the compound and the additional anti-cancer agent for the preparation of a medicament for the treatment of cancer.

26. A pharmaceutical composition comprising the compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt of the compound for use in cancer treatment.

27. A pharmaceutical composition comprising the compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt of the compound and an additional anti-cancer agent for use in cancer treatment.

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