Thiazolo[5,4-d]pyrimidine compounds, compositions containing the compounds, and methods of use thereof

Abstract

Compounds, compositions, and their uses in treating proliferative diseases or conditions, where, for example, the proliferative disease or disorder is associated with RAF gene mutations and / or RAS gene mutations. The compounds disclosed have the following formula I or a pharmaceutically acceptable salt and / or solvate thereof, wherein R 1 , R 2 , X 1 , X 2 , X 3 and X 4 is as defined herein. JPEG2025535134000151.jpg63170

Description

[Technical Field]

[0001] The present disclosure relates generally to thiazolo[5,4-d]pyrimidine compounds, pharmaceutical compositions containing the compounds, and their use in the treatment and prevention of diseases characterized by dysregulation of the RAS-ERK pathway (e.g., cancer, rasopathy). [Background technology]

[0002] The RAS-RAF-MEK-ERK (RAS (rat sarcoma): rat sarcoma; RAF (rapidly accelerated fibrosarcoma): rapidly accelerated fibrosarcoma; MEK (mitogen-activated protein kinase): mitogen-activated protein kinase; ERK (extracellular signal-regulated kinase): extracellular signal-regulated kinase) signaling pathway (hereinafter referred to as the RAS-ERK pathway) plays a key role in transmitting growth signals generated by growth factor receptors from the cell membrane to the nucleus. This pathway is dysregulated in the majority of cancers by the activation of receptor tyrosine kinases (RTKs) (e.g., ERBB1, ERBB2, FLT3, RET, KIT) or the activation or inactivation of RAS regulators (SOS1 and NF1), as well as by constitutively activating mutations in the RAS gene (H-, K-, NRAS; 30% of cancers) and the BRAF gene (8% of cancers). The prevalence of KRAS mutations is particularly high in pancreatic cancer (over 90%), colorectal cancer (50%), and lung cancer (30%), whereas BRAF mutations are found at particularly high frequencies in melanoma (70%), thyroid cancer (40%), and colorectal cancer (10%) (mutation frequencies based on COSMIC (Catalogue Of Somatic Mutations In Cancer; Wellcome Trust Sanger Institute) release v95, November 24, 2021).

[0003] RAS proteins are small GTPases that transmit extracellular growth signals to intracellular effectors to regulate important processes such as cell differentiation, proliferation, and survival (Nat. Rev. Cancer 2003, 3, 459). Following RTK stimulation, physiological activation of RAS occurs at the plasma membrane, resulting in the loading of GTP onto the GTPase and its activation. Activated RAS interacts with and activates a series of effector molecules, with RAF kinases being the most important RAS interactor in the context of cancer development (Nature Rev. Drug Discov. 2014, 13, 828). Oncogenic mutations at glycine 12, glycine 13, or glutamine 61 in RAS isoforms cause aberrant and constitutive signaling in human cancers (Nat. Rev. Cancer 2003, 3, 459) (COSMIC release v95, November 24, 2021).

[0004] Downstream of RAS, mammalian cells express three RAF paralogs (ARAF, BRAF, and CRAF), which share a conserved C-terminal kinase domain (KD) and an N-terminal regulatory region (NTR) containing a RAS-binding domain (RBD) (Nat. Rev. Mol. Cell Biol. 2015, 16, 281). In unstimulated cells, RAF proteins are sequestered in the cytoplasm as monomers. Binding of activated GTP-bound RAS to the RBD induces membrane anchoring of RAF kinases (Nat. Rev. Mol. Cell Biol. 2015, 16, 281). Concomitantly, RAF proteins undergo side-to-side dimerization of the kinase domain and catalytic activation (Nature 2009, 461, 542). Activated RAF proteins transmit signals through a phosphorylation cascade from RAF to MEK and then from MEK to ERK, leading to ERK phosphorylation of an array of substrates that trigger cell-specific responses (Nat. Rev. Mol. Cell Biol. 2020, Oct., 21(10), 607).

[0005] Activating mutations in RAF isoforms have so far been mostly restricted to the BRAF gene, but rare mutations have been observed in ARAF and CRAF, highlighting the functional importance of these isoforms (COSMIC release v95, November 24). th The most common cancer mutation in BRAF is a valine to glutamic acid substitution at position 600 (BRAF V600E(V600E allele) enhances BRAF activity by stabilizing its active form (Cell 2004, 116, 855). Apart from the V600E allele, various mutations occur at other residues (e.g., G466V, D594G, etc.) that cause increased RAF signaling through various mechanisms (Nat. Rev. Mol. Cell Biol. 2015, 16, 281). These have been classified into three major classes (1-3) depending on the level of dependence on RAS activity and RAF dimerization (Nature 2017 Aug 10, 548(7666), 234-238). The crucial role of wild-type BRAF and CRAF in mediating RAS-driven oncogenesis by stimulating ERK signaling has been widely validated (Cancer Cell 2011, 19, 652; Cancer Discov. 2012, 2, 685; Nat. Commun. 2017, 8, 15262). Thus, tumor cells rely on elevated and sustained signaling of the RAS-ERK pathway through activation of RAS and RAF, providing strong support for the concept of targeting RAF family kinases in cancer.

[0006] To address existing medical needs, a broad set of ATP-competitive RAF inhibitors has been developed over the past decade (Nat.Rev.Cancer 2017, 17, 76), primarily targeting the most common RAS-independent BRAF mutations (BRAF V600E ), leading to the development and FDA approval of sulfonamide derivatives, such as vemurafenib and dabrafenib. Some of these RAF inhibitors target recurrent BRAF V600E It has shown remarkable efficacy against metastatic melanoma with the BRAF allele and is approved for treating this patient population (N. Engl. J. Med. 2011, 364, 2507; Lancet 2012, 380, 358). V600EClinical responses against BRAF-dependent melanoma are due to potent, ATP-competitive inhibition of the monomeric form of this specific dimerization-independent BRAF mutant protein (Cancer Cell 2015, 28, 370). Unfortunately, acquired resistance to these agents inevitably develops, often due to reactivation of the RAS-ERK pathway, in part through mechanisms that stimulate RAF dimerization. These include upregulation of RTK signaling, RAS mutations, and BRAF V600E This includes amplification or cleavage (Sci. Signal. 2010, 3, ra84; Nature 2010, 468, 973; Nature 2011, 480, 387; Nature Commun. 2012, 3, 724).

[0007] At the same time, tumors that exhibit RAS activity due to activating RAS mutations or enhanced RTK signaling but are otherwise BRAF wild-type may be involved in the development of BRAF V600E RAF inhibitors exhibit primary resistance to RAF inhibitors (Nature 2010, 464, 431). Conversely, RAF inhibitors have been shown to induce ERK signaling under conditions of elevated RAS activity, thereby promoting tumor cell proliferation (Nature 2010, 464, 431). This counterintuitive phenomenon, known as the paradoxical effect, is also observed in normal tissues that depend on physiological RAS activity and is the basis for some of the side effects seen with RAF inhibitors in melanoma patients, such as the development of new secondary tumors (e.g., squamous cell carcinoma and keratoacanthomas) (Nat. Rev. Cancer 2014, 14, 455). Consequently, BRAFV600E inhibitors are ineffective and even contraindicated in RAS-driven cancers. The underlying mechanism results from the ability of compounds to promote dimerization of RAF kinase domains in the presence of activated RAS (Nature, 2010, 464, 431). This event is not limited to BRAF but also involves other RAF family members, and is determined by the binding mode and affinity of the compound (Nat. Chem. Biol. 2013, 9, 428).

[0008] To circumvent the limitations of first-generation RAF inhibitors in RAS-mutant cancers, two strategies have recently been pursued. The first strategy is based on the observation that paradoxical ERK activation is a dose-dependent phenomenon, i.e., induction occurs at subsaturating inhibitor concentrations, but the pathway is suppressed at saturating concentrations when the compound occupies both protomers of the RAF dimer. Therefore, the first strategy focused on developing molecules with high binding affinity to all RAF paralogs to saturate RAF proteins at lower drug concentrations and thereby reduce the induction of paradoxical pathways (Bioorg. Med. Chem. Lett. 2012, 22, 6237; Cancer Res. 2013, 73, 7043; J. Med. Chem. 2015, 58, 4165; Cancer Cell 2017, 31, 466; J Med Chem. 2020, 63, 2013; Clin Cancer Res. 2021, 27, 2061; Nature 2021, 594, 418). However, these compounds retained potent RAF dimer induction and thus paradoxically stimulated RAS-ERK signaling, although with a smaller amplitude than previous generation RAF inhibitors. Although this class of compounds exhibits improved properties, they have recently been found to largely avoid ARAF isoforms, leading to activation of paradoxical pathways and the emergence of primary and acquired resistance in vitro and in the clinic (Clin Cancer Res. 2021, 27, 2061; Nature 2021, 594, 418). A second strategy consisted in designing compounds that conformationally bias the BRAF kinase domain toward an inactive state and thus paradoxically do not induce ERK signaling. This resulted in the "Paradox Breaker" (PB) molecule PLX8394, a derivative of PLX4032 / vemurafenib (Nature 2015, 526, 583). These molecules inhibit BRAF. V600E It retains high efficacy against BRAF V600EThis should prove useful for treating RAS-dependent melanoma. However, although PLX8394 does not induce ERK signaling in the RAS-mutant cell lines tested, it remains ineffective and not useful against RAS-mutant tumors. Summary of the Invention [Problem to be solved by the invention]

[0009] There remains a need for inhibitors that potently and consistently inhibit RAS-ERK signaling and cell proliferation in human tumor cells with various RAS and RAF genotypes. The development of such inhibitors is also highly desirable because it is important that they do not induce paradoxical pathways in various RAS-mutated tumor cell lines. [Means for solving the problem]

[0010] According to one aspect, the present technology relates to compounds of Formula I below, or a pharmaceutically acceptable salt or solvate thereof: [ka] where: R 1 is replaced or not replaced, OR 3 , S.R. 3 , NH2, NHR 3 , N(R 3 )2, C 3~8 Cycloalkyl, C 4~8 Heterocycloalkyl, C 6~10 Aryl and C 5~10 heteroaryl; R 2 substituted C aryl or C 5~10 Heteroaryl, substituted or unsubstituted, C 4~8 Heterocycloalkyl, and N(R 3 )2 is selected; R 3 is independently in each occurrence substituted or unsubstituted, C1~8 Alkyl, C 3~8 Cycloalkyl, C 4~8 Heterocycloalkyl, C 6~10 Aryl and C 5~10 heteroaryl; X 1 is a halogen atom or an electron-withdrawing group; X 2 is selected from H, a halogen atom and an electron-withdrawing group; X 3 and X 4 are H, halogen atoms, electron-withdrawing groups, and C 1~3 Alkyl, C 3~4 Cycloalkyl and OC 1~3 alkyl.

[0011] Compounds of formula I are also defined according to any of the embodiments (alone or in combination) and examples described throughout this specification.

[0012] According to another aspect, the present technology relates to a pharmaceutical composition for use as defined in any one of the foregoing embodiments, wherein the composition comprises a compound as defined herein together with a pharmaceutically acceptable carrier, diluent, or excipient.

[0013] In a further aspect, the technology relates to the use of the compounds defined herein for the treatment of a disease or disorder selected from a proliferative disease or disorder, a developmental anomaly caused by dysregulation of the RAS-ERK signaling cascade (RASopathy), or an inflammatory disease or immune system disorder.

[0014] The present technology also relates to a method for treating a disease or disorder selected from a proliferative disease or disorder, a developmental anomaly caused by dysregulation of the RAS-ERK signaling cascade (RASopathy), or an inflammatory disease or immune system disorder, comprising administering a compound as defined herein to a subject in need of such treatment.A method for inhibiting the abnormal proliferation of cells, comprising contacting the cells with a compound as defined herein, is also contemplated.

[0015] In one embodiment of the above-mentioned uses and methods, the disease or disorder is selected from neoplasms and developmental anomalies, such as diseases or disorders associated with RAF gene mutations (e.g., ARAF, BRAF, or CRAF), diseases or disorders associated with RAS gene mutations (e.g., KRAS), or diseases or disorders associated with both RAF gene mutations and RAS gene mutations. In one embodiment, the disease or disorder is associated with receptor tyrosine kinase mutations or amplifications (e.g., EGFR, HER2), or mutations in downstream regulators of the receptor RAS (e.g., SOS1 gain-of-function, NF1 loss-of-function).

[0016] For example, the disease or disorder is a neoplasm, such as a neoplasm selected from melanoma, thyroid cancer (e.g., papillary thyroid cancer), colorectal cancer, ovarian cancer, breast cancer, endometrial cancer, liver cancer, sarcoma, gastric cancer, pancreatic cancer, Barrett's adenocarcinoma, glioma (e.g., ependymoma), lung cancer (e.g., non-small cell lung cancer), head and neck cancer, acute lymphocytic leukemia, acute myeloid leukemia, non-Hodgkin's lymphoma, and hairy cell leukemia. For example, the neoplasm is selected from colon or colorectal cancer, lung cancer, pancreatic cancer, thyroid cancer, breast cancer, and melanoma. For example, any of the above uses and methods of the present invention comprises inhibiting the RAS-ERK signaling pathway without substantially inducing a paradoxical pathway.

[0017] Further objects and features of the compounds, compositions, methods and uses of the present invention will become apparent upon reading the non-limiting description in the following illustrative embodiments and examples section, which should not be construed as limiting the scope of the invention. [Brief explanation of the drawings]

[0018] [Figure 1] FIG. 1 shows representative IC50 inhibition dose-response curves for compounds described herein that do not induce paradoxical induction of pERK signaling (Ymin >-20%) in RAS-mutated HCT116 cells (Examples 44 and 122) and a compound (PLX4720; CAS No. 918505-84-7) that causes strong induction of the pathway (Ymin ∼-600%) in the same cell line. DETAILED DESCRIPTION OF THE INVENTION

[0019] All technical and scientific terms used herein have the same definitions as commonly understood by those skilled in the art to which this technology pertains. Nevertheless, definitions of some terms and expressions used are provided below. To the extent that the definitions of terms in publications, patents, and patent applications incorporated herein by reference contradict the definitions set forth herein, the definitions herein shall prevail. The section headings used herein are for organizational purposes only and are not to be construed as limiting the disclosed subject matter.

[0020] i.Definition

[0021] The chemical structures described herein are drawn according to conventional standards. Also, when an atom, such as a carbon atom, is drawn with incomplete valences, it is assumed that the valences are satisfied by one or more hydrogen atoms, even if not necessarily explicitly drawn. The hydrogen atoms should be assumed to be part of the compound.

[0022] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting. It should be noted that the singular forms "a," "an," and "the" include the plural forms as well unless the content clearly dictates otherwise. Thus, for example, reference to a composition containing "a compound" also refers to a mixture of two or more compounds. It should also be noted that the word "or" is generally used in its sense to include "and / or" unless the context clearly dictates otherwise. Furthermore, to the extent the words "including," "includes," "having," "has," "with," or variations thereof are used in either the description and / or claims, such words are intended to be inclusive in a manner similar to the word "comprising."

[0023] The words "about" or "approximately" mean within an acceptable error range for a particular value, as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, "about" can mean within one standard deviation or more than one standard deviation, in accordance with practice in the art. Alternatively, "about" can mean a range of up to 20%, preferably up to 10%, more preferably up to 5%, and even more preferably up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, preferably within 5-fold, and more preferably within 2-fold of a value. Where particular values ​​are described in the present specification and claims, unless otherwise specified, the word "about" meaning within an acceptable error range for the particular value should be assumed.

[0024] As used herein, the terms "compound," "compounds described herein," "compounds of the present application," "thiazolo[5,4-d]pyrimidine compounds," "thiazolopyrimidine compounds," and their equivalents refer to compounds described herein, e.g., compounds encompassed by Structural Formula I, optionally with reference to any applicable embodiment, and also include exemplary compounds, e.g., compounds of Examples 1-159, and, where applicable, pharmaceutically acceptable salts, solvates, esters, and prodrugs thereof. Where a zwitterionic form is possible, the compound may be depicted as its neutral form for practical purposes, but it is understood that the compound also includes the zwitterionic form. Embodiments herein may also exclude one or more of the compounds. Compounds may be identified by either their chemical structure or their chemical name. In the event of a conflict between the chemical structure and the chemical name, the chemical structure will control.

[0025] Unless otherwise stated, structures depicted herein are also meant to include, where applicable, all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational) forms of the structure, e.g., the R and S configurations for each asymmetric center. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the compounds of the present application are within the scope of this specification. Unless otherwise specified, therapeutic compounds also include all possible tautomers, if any, of an exemplified compound. The term also includes isotopically labeled compounds where one or more atoms have an atomic mass different from the atomic mass most abundant in nature. Examples of isotopes that may be incorporated within the compounds of the invention are 2 H(D), 3 H(T), 11 C. 13 C. 14 C. 15 N, 18 O. 17The compounds may also exist in unsolvated forms as well as solvated forms, such as the solvated forms described above, including hydrates. The compounds may also exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated herein and are intended to be within the scope of the present invention.

[0026] When a particular enantiomer is preferred, it may, in some embodiments, be provided substantially free of the corresponding enantiomer, or it may be enantiomerically enriched. "Enantiomerically enriched" means that the compound is significantly enriched in one enantiomer. In some embodiments, the compound is made up of at least about 90% by weight of the preferred enantiomer. In other embodiments, the compound is made up of at least about 95%, 98%, or 99% by weight of the preferred enantiomer. The preferred enantiomer may be isolated from a racemic mixture by any method known to those skilled in the art, such as high-pressure liquid chromatography (HPLC) on a chiral support, and the methods described above, including the formation and crystallization of chiral salts, or may be prepared by asymmetric synthesis.

[0027] The expression "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention that is suitable, within the scope of sound medical judgment, for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, and that is commensurate with a reasonable benefit / risk ratio. These pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al. describe these pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66:1-19 (1977). The salts can be prepared in situ during the final isolation and purification of the compounds of the present invention, or can be prepared separately by reacting a free base functional group of the compound with a suitable organic or inorganic acid (acid addition salts), or by reacting an acidic functional group of the compound with a suitable organic or inorganic base (base addition salts).

[0028] The term "solvate" refers to a physical association of one of the compounds of the present invention with one or more solvent molecules, such as those listed above, including water and non-aqueous solvent molecules. This physical association may involve hydrogen bonding. In some cases, a solvate will be capable of isolation, for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. The term "solvate" encompasses both solution-phase and isolatable solvates. Exemplary solvates include, but are not limited to, hydrates, hemihydrates, ethanolates, hemiethanolates, n-propanolates, isopropanolates, 1-butanolates, 2-butanolates, and solvates of other physiologically acceptable solvents, such as Class 3 solvents described in the International Conference on Harmonization (ICH), Guide for Industry, Q3C Impurities: Residual Solvents (1997). Thus, the compounds described herein also encompass their respective solvates and mixtures thereof.

[0029] As used herein, the phrase "pharmaceutically acceptable ester" refers to an ester of a compound formed by a process herein that is hydrolyzable in vivo, including those that readily decompose in the human body to leave the parent compound or a salt thereof. Suitable ester groups include, for example, compounds derived from pharmaceutically acceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic, cycloalkanoic, and alkanedioic acids, where each alkyl or alkenyl moiety advantageously has six or fewer carbon atoms. Examples of specific esters include, but are not limited to, formates, acetates, propionates, butyrates, acrylates, and ethylsuccinates of hydroxyl groups, and alkyl esters of acidic groups. Other ester groups include sulfonates or sulfates.

[0030] As used herein, the phrase "pharmaceutically acceptable prodrug" refers to a prodrug of a compound formed by a process herein, which prodrug is suitable, within the scope of sound medical judgment, for use in contact with the tissues of humans and lower animals, is not associated with undue toxicity, irritation, allergic response, or the like, and is commensurate with a reasonable benefit / risk ratio and is effective for their intended use. As used herein, "prodrug" means a compound that is convertible in vivo by metabolic means (e.g., hydrolysis) to provide any compound depicted by a formula herein.

[0031] Abbreviations may also be used throughout this specification; unless otherwise specified, such abbreviations are intended to have the meaning generally understood in the art. Examples of such abbreviations are Me (methyl), Et (ethyl), Pr (propyl), i-Pr (isopropyl), Bu (butyl), t-Bu (tert-butyl), i-Bu (isobutyl), s-Bu (sec-butyl), c-Bu (cyclobutyl), Ph (phenyl), Bn (benzyl), Bz (benzoyl), CBz or Cbz or Z (carbobenzyloxy), Boc or BOC (tert-butoxycarbonyl), and Su or Suc (succinimide). For greater certainty, additional definitions of certain abbreviations are also included in the introduction to the "Examples" section.

[0032] The number of carbon atoms in the hydrocarbyl substituent is indicated by the prefix "C x ~C y " or "C x~y " where x is the minimum number of carbon atoms in the substituent and y is the maximum number of carbon atoms in the substituent. However, the prefix "C x ~C y " or "C x~y " is associated with a group that by definition incorporates one or more heteroatoms (e.g., heterocycloalkyl, heteroaryl, etc.), x and y define the minimum and maximum number of atoms in the ring, respectively, including carbon atoms and the one or more heteroatoms.

[0033] As used herein, the term "alkyl" refers to a saturated, straight- or branched-chain hydrocarbon moiety typically containing from 1 to 20 carbon atoms. For example, "C 1~8 Alkyl" contains 1 to 8 carbon atoms. Examples of alkyl moieties include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl, heptyl, octyl, and the like.

[0034] As used herein, the term "alkenyl" refers to a straight or branched chain hydrocarbon moiety containing one or more double bonds and typically containing from 2 to 20 carbon atoms. For example, "C 2~8 Alkenyl" contains 2 to 8 carbon atoms. Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, l-methyl-2-buten-l-yl, heptenyl, octenyl, and the like.

[0035] As used herein, the term "alkynyl" refers to a straight or branched chain hydrocarbon radical containing one or more triple bonds and typically containing from 2 to 20 carbon atoms. For example, "C 2~8 Alkynyl" contains from 2 to 8 carbon atoms. Representative alkynyl groups include, but are not limited to, for example, ethynyl, 1-propynyl, 1-butynyl, heptynyl, octynyl, and the like.

[0036] As used herein, the terms "cycloalkyl," "alicyclic," "carbocycle," "carbocyclic," and equivalents refer to groups containing monocyclic or polycyclic saturated or partially unsaturated (non-aromatic) carbocyclic rings, such as those described above, including spiro (sharing one atom), fused (sharing at least one bond), or bridged (sharing two or more bonds) carbocyclic ring systems, having 3 to 15 ring members. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopenten-1-yl, cyclopenten-2-yl, cyclopenten-3-yl, cyclohexyl, cyclohexen-1-yl, cyclohexen-2-yl, cyclohexen-3-yl, cycloheptyl, bicyclo[4,3,0]nonanyl, norbornyl, and the like. The term "cycloalkyl" encompasses both unsubstituted and substituted cycloalkyl groups. For example, the term "C3-n cycloalkyl" refers to a cycloalkyl group having three or more, and the indicated "n" carbon atoms in its ring structure. Unless the number of carbons is otherwise specified, "lower cycloalkyl" groups as used herein have at least three, and no more than eight, carbon atoms in their ring structure.

[0037] As used herein, the terms "heterocycle," "heterocycloalkyl," "heterocyclyl," "heterocyclic radical," and "heterocyclic ring" are used interchangeably and refer to a chemically stable 3- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocyclic moiety that is saturated or partially unsaturated and has, in addition to carbon atoms, one or more, preferably one to four, heteroatoms as defined above. When used in reference to a ring atom of a heterocycle, the term "nitrogen atom" includes substituted nitrogen atoms. For example, in a saturated or partially unsaturated ring having one to three heteroatoms selected from oxygen, sulfur, or nitrogen atoms, the nitrogen atom may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR (as in N-substituted pyrrolidinyl). A heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a chemically stable structure, and any of the ring atoms can be optionally substituted.Examples of heterocycloalkyl groups are 1,3-dioxolanyl, pyrrolidinyl, pyrrolidonyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydrodithienyl, tetrahydrothienyl, thiomorpholino, thioxanyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepaniyl, and the like. Examples include, but are not limited to, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, 3-azabicyclo[3,1,0]hexanyl, 3-azabicyclo[4,1,0]heptanyl, quinolidinyl, quinuclidinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, and the like. Heterocyclyl groups also include groups in which a heterocycle is fused to one or more aryl, heteroaryl, or cycloaliphatic rings, such as indolinyl, 3H-indolyl, chromanyl, chromenyl, phenanthridinyl, 2-azabicyclo[2.2.1]heptanyl, octahydroindolyl, or tetrahydroquinolinyl, where the moiety or point of attachment is on the heterocyclyl ring. Heterocyclyl groups can be monocyclic or bicyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted by a heterocyclyl, where the alkyl and heterocyclyl portions independently may be optionally substituted. For example, the term "C. 3~n "Heterocycloalkyl" refers to a heterocycloalkyl group having three or more, and the indicated "n" atoms in the ring structure, including carbon atoms and heteroatoms.

[0038] As used herein, the term "partially unsaturated" refers to a ring moiety that contains at least one double or triple bond between ring atoms, but that is not aromatic. The term "partially unsaturated" is intended to include rings with multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties, as defined herein.

[0039] The term "aryl," whether used alone or as part of a larger moiety, such as "aralkyl," "aralkoxy," "aryloxy," or "aryloxyalkyl," refers to an aromatic group having 4n+2 (where n is an integer from 1 to 3) conjugated π (pi) electrons in a monocyclic moiety or a bicyclic or tricyclic fused ring system having a total of 6 to 15 ring members, where at least one ring in the system is aromatic and each ring in the system contains 3 to 7 ring members. The term "aryl" may be used interchangeably with the term "aryl ring." In certain embodiments herein, "aryl" refers to an aromatic ring system, including, but not limited to, phenyl, biphenyl, naphthyl, azulenyl, anthracyl, and the like, which may bear one or more substituents. The term "aralkyl" or "arylalkyl" refers to an alkyl residue attached to an aryl ring. Examples of aralkyls include, but are not limited to, benzyl, phenethyl, and the like. Also included within the scope of the term "aryl," as used herein, are aromatic rings fused to one or more non-aromatic rings, such as indanyl, indenyl, phthalimidyl, naphthimidyl, fluorenyl, phenanthridinyl, or tetrahydronaphthyl. For example, the term "C 6~n "Aryl" refers to aryl groups having 6 or more atoms in the ring structure, with the indicated "n" number of atoms.

[0040] The term "heteroaryl," when used alone or as part of a larger moiety, such as "heteroaralkyl" or "heteroaralkoxy," refers to an aromatic group having 4n+2 (where n is an integer from 1 to 3) conjugated π (pi) electrons (e.g., having 5 to 18 ring atoms, preferably 5, 6, or 9 ring atoms; having 6, 10, or 14 π electrons shared in the cyclic array) and having 1 to 5 heteroatoms in addition to carbon atoms. The term "heteroatom" includes, but is not limited to, a nitrogen, oxygen, or sulfur atom, and includes any oxidized form of a nitrogen or sulfur atom and any quaternized form of a basic nitrogen atom. A heteroaryl may be a single ring or two or more fused rings. As used herein, the term "heteroaryl" also includes groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclic rings, where the moiety or point of attachment is on the heteroaromatic ring.Non-limiting examples of heteroaryl groups include thienyl, furanyl (furyl), pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, indolyl, 3H-indolyl, isoindolyl, indolizinyl, benzothienyl (benzothiophenyl), benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzoxazolyl, benzothiazolyl, benzotriazolyl, pyrrolopyridinyl (e.g., pyrrolo[3,2-b] or pyrrolo[3,2-c]pyridinyl), Heteroaryl groups include quinolyl (e.g., pyrazolo[1,5-a]pyridinyl), furopyridinyl, purinyl, imidazopyrazinyl (e.g., imidazo[4,5-b]pyrazinyl), quinolyl (quinolinyl), isoquinolyl (isoquinolinyl), quinolonyl, isoquinolonyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, naphthyridinyl, and pteridinylcarbazolyl, acridinyl, phenanthridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-l,4-oxazin-3(4H)-one. Heteroaryl groups may be monocyclic or bicyclic. Heteroaryl groups include rings that are optionally substituted. The term "heteroaralkyl" refers to an alkyl group substituted by a heteroaryl, where the alkyl and heteroaryl portions independently are optionally substituted. Examples include, but are not limited to, pyridinylmethyl, pyrimidinylethyl, and the like. For example, the term "C. 5~n "Heteroaryl" refers to heteroaryl groups having five or more, and the indicated "n" atoms in the ring structure, including carbon atoms and heteroatoms.

[0041] As described herein, the compounds herein may contain "optionally substituted" moieties. In general, the term "substituted," whether preceded by the term "optionally," means that one or more hydrogen atoms of the specified moiety are replaced with a suitable substituent. Unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents may be the same or different at each position. Combinations of substituents contemplated herein preferably result in the formation of chemically stable or chemically feasible compounds. As used herein, the term "chemically stable" refers to compounds that remain substantially unchanged when subjected to conditions that enable their preparation, detection, and, in some embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein.

[0042] The term "halo" means a halogen atom, ie, a fluorine, chlorine, bromine or iodine atom, preferably a fluorine or chlorine atom.

[0043] The term "optionally substituted" refers to a group that is substituted or unsubstituted by independently replacing one, two, or more of the hydrogen atoms on the group with a substituent, such as those listed above, including, but not limited to, the following: F, CI, Br, I, OH, COH, alkoxy, oxo, thioxo, NO, CN, CF, NH, NH-alkyl, NH-alkenyl, NH-alkynyl, NH-cycloalkyl, NH-aryl, NH-heteroaryl, NH-heterocycle, dialkylamino, diarylamino, diheteroarylamino, O-aryl, O-aryl- ... alkyl, O-alkenyl, O-alkynyl, O-cycloalkyl, O-aryl, O-heteroaryl, O-haloalkyl, O-heterocycle, C(O)alkyl, C(O)alkenyl, C(O)alkynyl, C(O)cycloalkyl, C(O)aryl, C(O)heteroaryl, C(O)heterocycloalkyl, CO2alkyl, CO2alkenyl, CO2alkynyl, CO2cycloalkyl, CO2aryl, CO2heteroaryl, CO2heterocycloalkyl, OC(O)alkyl, OC(O)alkenyl, OC(O)alkynyl, OC(O)cycloalkyl alkyl, OC(O)aryl, OC(O)heteroaryl, OC(O)heterocycloalkyl, C(O)NH2, C(O)NHalkyl, C(O)NHalkenyl, C(O)NHalkynyl, C(O)NHcycloalkyl, C(O)NHaryl, C(O)NHheteroaryl, C(O)NHheterocycloalkyl, OCO2alkyl, OCO2alkenyl, OCO2alkynyl, OCO2cycloalkyl, OCO2aryl, OCO2heteroaryl, OCO2heterocycloalkyl, OC(O)NH2, OC(O)NHalkyl, OC(O)NHalkynyl nyl, OC(O)NHalkynyl, OC(O)NHcycloalkyl, OC(O)NHaryl, OC(O)NHheteroaryl, OC(O)NHheterocycloalkyl, NHC(O)alkyl, NHC(O)alkenyl, NHC(O)alkynyl, NHC(O)cycloalkyl, NHC(O)aryl, NHC(O)heteroaryl, NHC(O)heterocycloalkyl, NHCO2alkyl, NHCO2alkenyl, NHCO2alkynyl, NHCO2cycloalkyl, NHCO2aryl, NHCO2heteroaryl, NHCO2heterocycloalkyl,NHC(O)NH2, NHC(O)NH alkyl, NHC(O)NH alkenyl, NHC(O)NH alkynyl, NHC(O)NH cycloalkyl, NHC(O)NH aryl, NHC(O)NH heteroaryl, NHC(O)NH heterocycloalkyl, NHC(S)NH2, NHC(S)NH alkyl, NHC(S)NH alkenyl, NHC(S)NH alkynyl, NHC(S)NH cycloalkyl, NHC(S)NH aryl, NHC(S)NH heteroaryl, NHC(S)NH heterocycloalkyl, NHC(NH)NH2, NHC(NH NH alkyl, NHC(NH)NH alkenyl, NHC(NH)NH alkynyl, NHC(NH)NH cycloalkyl, NHC(NH)NH aryl, NHC(NH)NH heteroaryl, NHC(NH)NH heterocycloalkyl, NHC(NH) alkyl, NHC(NH) alkenyl, NHC(NH) alkynyl, NHC(NH) cycloalkyl, NHC(NH) aryl, NHC(NH) heteroaryl, NHC(NH) heterocycloalkyl, C(NH)NH alkyl, C(NH)NH alkenyl, C(NH)NH alkynyl, C(NH)NH cycloalkyl Cycloalkyl, C(NH)NHaryl, C(NH)NHheteroaryl, C(NH)NHheterocycloalkyl, P(O)(alkyl)2, P(O)(alkenyl)2, P(O)(alkynyl)2, P(O)(cycloalkyl)2, P(O)(aryl)2, P(O)(heteroaryl)2, P(O)(heterocycloalkyl)2, P(O)(Oalkyl)2, P(O)(OH)2, P(O)(Oalkenyl)2, P(O)(Oalkynyl)2, P(O)(Ocycloalkyl)2, P(O)(Oaryl)2, P(O)(Oheteroaryl)2, P(O )(Oheterocycloalkyl)2, S(O)alkyl, S(O)alkenyl, S(O)alkynyl, S(O)cycloalkyl, S(O)aryl, S(O)2alkyl, S(O)2alkenyl, S(O)2alkynyl, S(O)2cycloalkyl, S(O)2aryl, S(O)heteroaryl, S(O)heterocycloalkyl, SO2NH2, SO2NHalkyl, SO2NHalkenyl, SO2NHalkynyl, SO2NHcycloalkyl, SO2NHaryl, SO2NHheteroaryl, SO2NHheterocycloalkyl, NHSO2alkyl,NHSO2alkenyl, NHSO2alkynyl, NHSO2cycloalkyl, NHSO2aryl, NHSO2heteroaryl, NHSO2heterocycloalkyl, CH2NH2, CH2SO2CH3, alkyl, alkenyl, alkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, cycloalkyl, carbocyclic, heterocyclic, polyalkoxyalkyl, polyalkoxy, methoxymethoxy, methoxyethoxy, SH, S-alkyl, S-alkenyl, S-alkynyl, S-cycloalkyl, S-aryl, S-heteroaryl, S-heterocycloalkyl, or methylthiomethyl.

[0044] ii. compound

[0045] The recitation of a list of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of embodiments of a variable herein includes those embodiments as any single embodiment or in combination with any other embodiment or portion thereof. The recitation of embodiments herein includes those embodiments. Thus, the following embodiments, where applicable, occur alone or in combination.

[0046] The compounds of the present invention exhibit a thiazolo[5,4-d]pyrimidine core structure to which defined substituents are attached to achieve the beneficial activity of the product. Examples of thiazolopyrimidine compounds defined herein, or their pharmaceutically acceptable salts or solvates thereof, are represented by the following general formula I: [ka] where: R 1 is replaced or not replaced, OR 3 , S.R. 3 , NH2, NHR 3 , N(R 3 )2, C 3~8 Cycloalkyl, C4~8 Heterocycloalkyl, C 6~10 Aryl and C 5~10 heteroaryl, e.g., substituted or unsubstituted, SR 3 , N(R 3 )2, C 4~8 Heterocycloalkyl, C 6~10 Aryl and C 5~10 heteroaryl, preferably substituted or unsubstituted, C 6~10 Aryl or C 5~10 heteroaryl; R 2 substituted C aryl or C 5~10 Heteroaryl, substituted or unsubstituted, C 4~8 Heterocycloalkyl, and N(R 3 )2 is selected; R 3 is independently in each occurrence substituted or unsubstituted, C 1~8 Alkyl, C 3~8 Cycloalkyl, C 4~8 Heterocycloalkyl, C 6~10 Aryl and C 5~10 heteroaryl; X 1 is a halogen atom or an electron-withdrawing group; X 2 is selected from H, a halogen atom and an electron-withdrawing group; X 3 and X 4 are H, halogen atoms, electron-withdrawing groups, and C 1~3 Alkyl, C 3~4 Cycloalkyl and OC 1~3 alkyl.

[0047] For example, the electron-withdrawing group is selected from perhaloalkyl (e.g., CF3 or CCl3), CN, NO2, sulfonate, alkylsulfonyl (e.g., SO2Me or SO2CF3), alkylcarbonyl (e.g., C(O)Me), carboxylate, alkoxycarbonyl (e.g., C(O)OMe), and aminocarbonyl (e.g., C(O)NH2). In one embodiment, X 1 is Cl, and X 2 is F, or X 1 is F, and X 2 is H, or X 1 and X 2 are both F. In another embodiment, X 3 and X 4 are each H. In yet another embodiment, X 3 is F, and X 4 is H.

[0048] For example, the aminoarylsulfonamide moiety in Formula I may be designated L and is preferably selected from the following: [ka] where the dashed line (---) represents the bond that serves as the point of attachment between L and the remainder of the molecule.

[0049] In a further embodiment, R 2 substituted C aryl or C 5~10 Heteroaryl, for example, R 2 is F, Cl, Br, CN, NO2 and substituted or unsubstituted, C 1~3 Alkyl, C 3~4 Cycloalkyl or OC 1~3 and C6 aryl substituted with at least one group selected from alkyl. For example, R 2 is a group of the formula: [ka] where: R 4is H, F, Cl, Br, CN, and substituted or unsubstituted, C 1~3 Alkyl, C 3~4 Cycloalkyl or OC 1~3 alkyl, for example, R 4 is selected from H, F, Cl, Br, Me, Et, CN, CHF2 and CF3; R 5 is H, F, Cl, CN, and substituted or unsubstituted, C 1~3 Alkyl, C 3~4 Cycloalkyl or OC 1~3 alkyl, for example, R 5 is selected from H, F, Me, CF3, CN and Cl; R 6 is H, F, Cl, Br, NO2, NH2, and substituted or unsubstituted C 1~3 Alkyl, C 3~4 Cycloalkyl or OC 1~3 alkyl, for example, R 6 is H, F, Cl, Br, and substituted or unsubstituted C 1~3 Alkyl, C 3~4 Cycloalkyl or OC 1~3 alkyl, or R 6 is selected from H, F, Cl, Me, Et, and OMe; R 7 is H, F, Cl, and substituted or unsubstituted C 1~3 alkyl, for example, R 7 is selected from H, Me, F and Cl; R 8 H, F, and substituted or unsubstituted C 1~3 alkyl, for example, R 8 is selected from H, Me, and F; Or R 4 and R 5 Or R 5 and R 6 together with their adjacent carbon atoms form a substituted or unsubstituted carbocyclic or heterocyclic ring, provided that the heterocyclic ring (R 2) is not a benzoxazolinone; and (---) is R 2 represents the bond that serves as the point of attachment between and the rest of the molecule; where R 4 is H or F, then R 5 , R 6 , R 7 or R 8 is other than H or F; and where R 5 If is CN, then R 4 , R 6 , R 7 or R 8 At least one of is other than H.

[0050] In one embodiment, R 8 is H. In another embodiment, R 4 is selected from F, Cl, Et and Me, R 5 , R 7 and R 8 are each H, and R 6 is selected from H, Cl, Me and OMe. In a further embodiment, R 4 is selected from F, Cl and Me, R 6 , R 7 and R 8 are each H, and R 5 is selected from F and Cl.

[0051] In a further embodiment, R 4 is Cl, and substituted or unsubstituted C 1~3 alkyl (e.g., Me); R 5 is H, F, Cl, and substituted or unsubstituted C 1~3 alkyl (e.g., Me); R 6 is H, and substituted or unsubstituted OC 1~3 alkyl (e.g., OCH); and R 7 and R 8 are each H.

[0052] In yet another embodiment, R 4 is selected from H, Cl, Br and methyl; R 5 is selected from H, F and Cl; R 6 is selected from H, F, Cl, Me, and OMe; and R 7 and R 8 are each H.

[0053] In a further embodiment, R 4 is Cl, and substituted or unsubstituted C 1~3 alkyl, preferably R 4 is Cl or Me; R 5 is H, F, Cl, and substituted or unsubstituted C 1~3 alkyl (e.g., Me), preferably R 5 is F, Cl or Me; R 6 is H, F, Cl, substituted or unsubstituted C 1~3 Alkyl (e.g., Me), and substituted or unsubstituted OC 1~3 alkyl (e.g., OCH), preferably R 6 is H or F, or R 6 is Cl, or substituted or unsubstituted C 1~3 Alkyl, or substituted or unsubstituted OC 1~3 alkyl, or R 6 is CH3 or OCH3; and R 7 and R 8 are each H. In still other embodiments, R 6 is the substituted C 1~3 It is alkyl.

[0054] In another example, R 2 is a substituted C5 heteroaryl, such as a group of the formula: [ka] where: X 5 However, NH, NC1~3 Alkyl, NC 3~4 cycloalkyl, selected from O and S; R 9 , R 10 , R 11 are each independently H, F, Cl, CN, and substituted or unsubstituted, C 1~3 Alkyl, C 3~4 Cycloalkyl, C(O)OC 1~3 Alkyl or OC 1~3 alkyl, with the proviso that R 9 and R 11 one of which is H and the other is not H; and (---) is R 2 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

[0055] Alternatively, R 2 is a group of the formula: [ka] where: X 5 However, NH, NC 1~3 Alkyl, NC 3~4 cycloalkyl, selected from O and S; R 9 is F, Cl, CN, and substituted or unsubstituted, C 1~3 Alkyl, C 3~4 Cycloalkyl, C(O)OC 1~3 Alkyl or OC 1~3 alkyl; R 10 and R 12 are each independently H, F, Cl, CN, and substituted or unsubstituted, C 1~3 Alkyl, C 3~4 Cycloalkyl, C(O)OC 1~3 Alkyl or OC 1~3 alkyl; and (---) is R 2 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

[0056] In a preferred embodiment, R 9 and R 10 are each independently selected from F, Cl, CN, and substituted or unsubstituted, C 1~3 Alkyl, C 3~4 Cycloalkyl, C(O)OC 1~3 Alkyl or OC 1~3 alkyl, preferably Cl, and substituted or unsubstituted C 1~3 alkyl, more preferably R 9 and R 10 are both Cl. In another embodiment, X 5 is O or S, preferably S.

[0057] In another embodiment, R 2 is the substituted C 5~10 Heteroaryl, for example, a group of the formula: [ka] where: X 9 , X 10 , X 11 , X 12 and X 13 are independently selected from N and C, where X 9 , X 10 , X 11 , X 12 and X 13 at least one and not more than two of are N; and R 19 , R 20 , R 21 , R 22 and R 23 is H, F, Cl, Br, CN, NO2, NH2, and substituted or unsubstituted C 1~3 Alkyl, C 3~4 Cycloalkyl or OC 1~3 alkyl or their bonded X 9 , X 10 , X 11 , X 12 or X 13does not exist when N; where X 9 and X 13 At least one of is not N; where X 9 and X 13 when one of is N, the other is not N or CH; (---) is R 2 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

[0058] In another example, R 2 is a C5 heterocycloalkyl. For example, R 2 is a group of the formula: [ka] where: R 13 represents independently in each occurrence F, Cl, and substituted or unsubstituted, C 1~3 Alkyl, C 3~4 Cycloalkyl or C 1~3 selected from alkoxy; n is an integer selected from 0 to 8; or n is 2 to 8, and two R 13 together with their adjacent carbon atoms, C 3~4 forming a cycloalkyl; and (---) is R 2 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

[0059] In one embodiment, R 13 is at position 3. In another embodiment, R 13 is selected from F, Me, OMe, and CHOMe, and n is 1 or 2. For example, R 13 is a methoxy group at the 3-position, and n is 1.

[0060] In a further example, R 2 is N(R 3 )2. For example, R2 is N(R 3 )2, and R 3 is substituted or unsubstituted, C 1~8 Alkyl or C 3~8 cycloalkyl.

[0061] In yet another embodiment, the compound of formula I is a compound of formula II: or a pharmaceutically acceptable salt or solvate thereof. [ka] where R 1 , R 4 , R 5 and R 6 are each independently as defined above, preferably R 4 is selected from Cl, Br and methyl; R 5 is selected from H, F, Cl and methyl, and R 6 is selected from H, F, Cl, Me and OMe.

[0062] In a further embodiment, the compound of formula I is a compound of formula III below, or a pharmaceutically acceptable salt or solvate thereof: [ka] where R 1 , R 9 , R 10 , R 12 and X 5 are each independently as defined above.

[0063] Illustrative R 2 Groups are selected from groups B1 to B77 defined below.

[0064] [ka] JPEG2025535134000013.jpg217170JPEG2025535134000014.jpg63170

[0065] where (---) is R 2 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

[0066] In one embodiment, R 2 is selected from the groups B1 to B77, or preferably R 2 is selected from groups B1 to B6.

[0067] In one embodiment of the compound of formula I, R 1 is OR 3 or SR 3 For example, R 1 is SR 3 In various embodiments, R 3 is a substituted or unsubstituted C 1~8 Alkyl (e.g., C 1~3 alkyl).

[0068] In another embodiment, R 1 is a substituted or unsubstituted C aryl group. In another embodiment, R 1 is a substituted or unsubstituted C 4~6 Heterocycloalkyl groups. For example, R 1 are halogen atoms, OH, C 1~6 Alkyl and OC 1~6 C optionally substituted with one or two groups selected from alkyl 4~5 Heterocycloalkyl. For example, R 1 is an N-pyrrolidinyl group substituted with one or two groups selected from F and OH.

[0069] In another embodiment, R 1 is a substituted or unsubstituted C 5~6 In another embodiment, R is a heteroaryl group or a substituted or unsubstituted C heteroaryl group. 1is a substituted or unsubstituted group selected from thienyl, imidazolyl, pyrazolyl, triazolyl, thiazolyl, pyridyl, pyrimidinyl, indolyl, indazolyl, benzimidazolyl, benzotriazolyl, pyrrolopyridinyl (e.g., pyrrolo[3,2-b]pyridinyl or pyrrolo[3,2-c]pyridinyl), pyrazolopyridinyl (e.g., pyrazolo[1,5-a]pyridinyl), purinyl, imidazopyrazinyl (e.g., imidazo[4,5-b]pyrazinyl), and quinolyl (quinolinyl), preferably R 1 is a substituted or unsubstituted group selected from imidazolyl, pyrazolyl, triazolyl, indolyl, indazolyl, benzimidazolyl, benzotriazolyl, pyrrolopyridinyl (e.g., pyrrolo[3,2-b]pyridinyl or pyrrolo[3,2-c]pyridinyl), pyrazolopyridinyl (e.g., pyrazolo[1,5-a]pyridinyl), purinyl, and imidazopyrazinyl (e.g., imidazo[4,5-b]pyrazinyl), more preferably a substituted or unsubstituted group as described above attached to the thiazolopyrimidine core through a nitrogen atom.

[0070] R 1 Examples of include substituted or unsubstituted groups selected from: [ka]

[0071] where (---) is R 1 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

[0072] For example, R 1 is a substituted or unsubstituted group selected from: [ka]

[0073] where (---) is R 1represents the bond that serves as the point of attachment between the group and the rest of the molecule.

[0074] In one embodiment, R 1 OH, halogen atoms, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, OC 1~6 Alkyl, C 5~10 Heteroaryl, C 3~10 Cycloalkyl, C 4~10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 )2, SO2R 15 , SO2N(R 14 )2, N(R 16 )C(O)R 15 , N(R 16 )SO2R 15 , N(R 16 )C(O)N(R 14 )2, N(R 16 )SO2N(R 14 )2, N(R 14 )2, P(O)(R 15 )2, CH2C(O)R 15 , CH2C(O)N(R 14 )2, CH2SO2R 15 , CH2SO2N(R 14 )2, CH2N(R 16 )C(O)R 15 , CH2N(R 16 )SO2R 15 , CH2N(R 16 )C(O)N(R 14 )2, CH2N(R 16 )SO2N(R 14 )2 and CH2N(R 14 ) one of the above groups further substituted with at least one substituent selected from where: R 14 independently in each occurrence, H, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~10 Cycloalkyl, C4~10 Heterocycloalkyl, C6 aryl, and C 5~10 heteroaryl, or two R 14 together with their adjacent nitrogen atoms to form C 4~10 forming a heterocycloalkyl group; R 15 independently in each occurrence, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~10 Cycloalkyl, C6 aryl, and C 5~10 heteroaryl; and R 16 independently in each occurrence, H, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~10 Cycloalkyl, C6 aryl, and C 5~10 heteroaryl; where R 1 (R 14 , R 15 and R 16 The alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl groups contained within (including the definition of) may be further optionally substituted.

[0075] In another embodiment, R 1 is a group of the formula: [ka] where: R 17 However, H, OH, halogen atoms, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, OC 1~6 Alkyl, C 5~10 Heteroaryl, C 3~10 Cycloalkyl, C 4~10 Heterocycloalkyl, C(O)R 15 , C(O)N(R14 )2, SO2R 15 , SO2N(R 14 )2, N(R 16 )C(O)R 15 , N(R 16 )SO2R 15 , N(R 16 )C(O)N(R 14 )2, N(R 16 )SO2N(R 14 )2, N(R 14 )2, P(O)(R 15 )2, CH2C(O)R 15 , CH2C(O)N(R 14 )2, CH2SO2R 15 , CH2SO2N(R 14 )2, CH2N(R 16 )C(O)R 15 , CH2N(R 16 )SO2R 15 , CH2N(R 16 )C(O)N(R 14 )2, CH2N(R 16 )SO2N(R 14 )2 and CH2N(R 14 )2 is selected; R 27 However, H, OH, halogen atoms, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, OC 1~6 Alkyl, C 5~10 Heteroaryl, C 3~10 Cycloalkyl, C 4~10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 )2, SO2R 15 , SO2N(R 14 )2, N(R 16 )C(O)R 15 , N(R 16 )SO2R 15 , N(R 16 )C(O)N(R 14 )2, N(R 16 )SO2N(R 14 )2, N(R 14 )2, P(O)(R 15)2, CH2C(O)R 15 , CH2C(O)N(R 14 )2, CH2SO2R 15 , CH2SO2N(R 14 )2, CH2N(R 16 )C(O)R 15 , CH2N(R 16 )SO2R 15 , CH2N(R 16 )C(O)N(R 14 )2, CH2N(R 16 )SO2N(R 14 )2 and CH2N(R 14 ) 2, preferably H, a halogen atom (e.g., F), optionally substituted C 1~6 alkyl, or optionally substituted OC 1-6 alkyl; X 6 is N or CH; and X 7 is N, and R 18 is not present; or X 7 is C, and R 18 But C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, OC 1~6 Alkyl, C 5~10 Heteroaryl, C 3~10 Cycloalkyl, C 4~10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 )2, SO2R 15 , SO2N(R 14 )2, N(R 16 )C(O)R 15 , N(R 16 )SO2R 15 , N(R 16 )C(O)N(R 14 )2, N(R 16 )SO2N(R 14 )2, N(R 14 )2, P(O)(R 15 )2, CH2C(O)R 15 , CH2C(O)N(R 14)2, CH2SO2R 15 , CH2SO2N(R 14 )2, CH2N(R 16 )C(O)R 15 , CH2N(R 16 )SO2R 15 , CH2N(R 16 )C(O)N(R 14 )2, CH2N(R 16 )SO2N(R 14 )2 and CH2N(R 14 )2 is selected; where R 14 , R 15 and R 16 is as defined above; where R 1 (R 14 , R 15 , R 16 , R 17 and R 18 The alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, or heteroaryl contained in the definition of (which includes the definition of) may be further optionally substituted; and where (---) is R 1 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

[0076] In another embodiment, R 1 is the basis of the formula: [ka] where: X 15 , X 16 , X 17 , and X 18 are independently O, N, S and CR 17 where R 17 is as defined above; where X 15 , X 16 , X 17 and X 18 not more than two of are O, N, or S; and where (---) is R1 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

[0077] In one embodiment, the compound of formula I is a compound of formula IV or V below, or a pharmaceutically acceptable salt or solvate thereof. [ka] [ka] where R 4 , R 5 , R 6 , R 17 , R 18 , R 27 , X 6 , X 7 , X 15 , X 16 , X 17 and X 18 are each independently as defined herein, preferably R 4 is selected from Cl, Br and methyl; R 5 is selected from H, F, Cl and methyl; R 6 is selected from H, Cl, F, Me and OMe.

[0078] In further embodiments, the compound of formula I is a compound of formula VI or formula VII below, or a pharmaceutically acceptable salt or solvate thereof. [ka] [ka]

[0079] where R 9 , R 10 , R 12 , R 17 , R 18 , R 27 , X 5 , X 6 , X 7 , X15 , X 16 , X 17 and X 18 are each independently as defined herein.

[0080] In one embodiment of the above formula, X 6 is N. In another embodiment, X 6 is CH.

[0081] In another embodiment, X 7 is N and R 17 H, OH, halogen atoms, CN, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, OC 1~6 Alkyl, C 5~10 Heteroaryl, C 3~10 Cycloalkyl, C 4~10 Heterocycloalkyl, C(O)R 15 ,C(O)N(R 14 )2, SO2R 15 , SO2N(R 14 )2, N(R 16 )C(O)R 15 , N(R 16 )SO2R 15 , N(R 16 )C(O)N(R 14 )2, N(R 16 )SO2N(R 14 )2, N(R 14 )2, P(O)(R 15 )2, CH2C(O)R 15 , CH2C(O)N(R 14 )2, CH2SO2R 15 , CH2SO2N(R 14 )2, CH2N(R 16 )C(O)R 15 , CH2N(R 16 )SO2R 15 , CH2N(R 16 )C(O)N(R 14 )2, CH2N(R 16 )SO2N(R 14 )2 and CH2N(R 14)2 and R 18 does not exist, where R 14 , R 15 , R 16 or R 17 The alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl or heteroaryl in the formula (I) may be optionally further substituted, and preferably R 17 is C 1~6 Alkyl, C 5~10 Heteroaryl, C 4~10 Heterocycloalkyl, N(R 14 )2, N(R 16 )C(O)R 15 , N(R 16 )SO2R 15 , C(O)N(R 14 )2 and SO2N(R 14 )2, where R 14 , R 15 , R 16 or R 17 The alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, or heteroaryl in the formula (I) may be further optionally substituted. For example, R 17 is H, F, NH2 and optionally substituted, C 5~10 Heteroaryl or C 4~10 heterocycloalkyl, preferably R 17 is an optionally substituted C 5~10 Heteroaryl or C 4~10 It is heterocycloalkyl.

[0082] In a further embodiment, R 17 is an optionally substituted C 4~10 heterocycloalkyl, wherein the heterocycloalkyl may be monocyclic or bicyclic and contains 1 to 3 heteroatoms, preferably wherein X 7 is N. In a preferred embodiment, the heterocycloalkyl is, for example, F, OH, oxo, CN, C 1~4 Alkyl and OC 1~4alkyl, wherein the C 1~4 Alkyl is (e.g., F, OH, OC 1~3 For example, the heterocycloalkyl may be selected from optionally substituted piperidine, piperazine, thiomorpholine, and morpholine groups, or bicyclic structures (bridged or spiro) containing a piperidine, piperazine, thiomorpholine, or morpholine ring.

[0083] In a further embodiment, X 7 is C, for example, X 7 is C and R 18 is C 1~6 Alkyl, C 5~10 Heteroaryl, C 3~10 Cycloalkyl, C 4~10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 )2, SO2R 15 , SO2N(R 14 )2, N(R 16 )C(O)R 15 , N(R 16 )SO2R 15 , N(R 16 )C(O)N(R 14 )2, N(R 16 )SO2N(R 14 )2, N(R 14 )2, P(O)(R 15 )2, CH2C(O)R 15 , CH2C(O)N(R 14 )2, CH2SO2R 15 , CH2SO2N(R 14 )2, CH2N(R 16 )C(O)R 15 , CH2N(R 16 )SO2R 15 , CH2N(R 16 )C(O)N(R 14 )2, CH2N(R 16 )SO2N(R 14 )2 and CH2N(R 14 )2, where R14 , R 15 , R 16 or R 18 The alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl or heteroaryl in the formula (I) may be optionally further substituted, and preferably R 18 is C(O)N(R 14 )2, SO2R 15 and SO2N(R 14 )2. In a subclass of these embodiments, R 17 is H, halogen atoms, OH, C 1~6 Alkyl, N(R 14 )2, and optionally substituted C 5~10 For example, R 17 is H, F, NH2 and optionally substituted C 5~10 Heteroaryl is preferably selected from H, F or NH2.

[0084] In yet another embodiment, R 14 is independently in each occurrence H, optionally substituted C 1~6 Alkyl, optionally substituted C 3~10 Cycloalkyl, optionally substituted C 4~10 Heterocycloalkyl and optionally substituted C 5~6 heteroaryl, or two R 14 together with their adjacent nitrogen atoms, optionally substituted C 4~10 It forms a heterocycloalkyl group.

[0085] In another embodiment, R 17 is N(R 14 )2, wherein the R 14 together with their adjacent nitrogen atoms, C 4~10 and form a heterocycloalkyl group, wherein the heterocycloalkyl may be monocyclic or bicyclic and contains 1 to 3 heteroatoms, preferably wherein X 7is N. In a preferred embodiment, the heterocycloalkyl is, for example, F, OH, oxo, CN, C 1~4 Alkyl and OC 1~4 alkyl, wherein the C 1~4 Alkyl is (e.g., F, OH, OC 1~3 For example, the heterocycloalkyl is selected from optionally substituted piperidine, piperazine, thiomorpholine, and morpholine groups, or bicyclic structures (bridged or spiro) containing a piperidine, piperazine, thiomorpholine, or morpholine ring.

[0086] In another example, R 1 is selected from the following: [ka]

[0087] where R 14 , R 17 and R 27 is as defined herein, and (---) represents R 1 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

[0088] In a further example, R 1 is selected from the following: [ka]

[0089] where R 14 , R 17 and R 27 is as defined herein, and (---) represents R 1 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

[0090] Each substituent R 1 (A group), R 2Further subembodiments, including (group B) and group L, are also presented in the Examples section. Exemplary combinations are also described in Tables 3 and 4 below. Representative preferred compounds of Examples 1 to 159 are also described herein.

[0091] More specifically, the exemplary R 1 The groups are selected from the groups A1 to A550 defined below.

[0092] [ka] JPEG2025535134000026.jpg210170JPEG2025535134000027.jpg231170JPEG2025535134000028.jpg21 1170JPEG2025535134000029.jpg232170JPEG2025535134000030.jpg216170JPEG2025535134000031.j pg225170JPEG2025535134000032.jpg231170JPEG2025535134000033.jpg204170JPEG20255351340000 34.jpg201170JPEG2025535134000035.jpg226170JPEG2025535134000036.jpg213170JPEG20255351340 00037.jpg202170JPEG2025535134000038.jpg205170JPEG2025535134000039.jpg201170JPEG2025535 134000040.jpg207170JPEG2025535134000041.jpg215170JPEG2025535134000042.jpg217170JPEG202 5535134000043.jpg222170JPEG2025535134000044.jpg232170JPEG2025535134000045.jpg229170JPE G2025535134000046.jpg222170JPEG2025535134000047.jpg230170JPEG2025535134000048.jpg153170

[0093] where (---) is R 1 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

[0094] In one embodiment, R 1 is selected from the groups A1 to A550, or R 1 is selected from the groups A1 to A3, A8, A19, A20, A22, A23, A25, A28, A29, A32 to A39, A60, A63 to A66, A69, A72 to A78, A81 to A83, A86, A89, A96, A100, A101, A104, A105, A109 to A111, A113, A115, A118, A121 to A123, A127 and A132, for example, R 1 is selected from the groups A1 to A3, A8, A19, A22, A25, A28, A29, A32, A36, A37, A64, A67, A74, A77, A78, A82, A83, A89, A96, A109, A110 and A111.

[0095] In one embodiment, R 1 is selected from the groups A1 to A3, A8, A19, A20, A22, A23, A25, A28, A29, A32 to A39, A60, A63 to A66, A69, A72 to A78, A81 to A83, A86, A89, A93, A96, A100, A101, A104, A105, A109 to A111, A113, A115, A118, A121 to A123, A127 and A132, for example, R 1 is selected from the groups A1 to A3, A8, A19, A22, A25, A28, A29, A32, A36, A37, A64, A67, A74, A77, A78, A82, A83, A89, A93, A96, A109, A110 and A111.

[0096] The following embodiments are R groups that can be combined to prepare compounds of formula I: 1 (A1~A550) Group, R 2 The combinations of (B1 to B77) groups and L (L1 to L4) groups are shown below. A1-L-B1;A1-L-B2;A1-L-B3;A1-L-B4~B75;A1-L-B76;A1-L-B77; A2-L-B1;A2-L-B2;A2-L-B3;A2-L-B4~B75;A2-L-B76;A2-L-B77; A3-L-B1;A3-L-B2;A3-L-B3;A3-L-B4~B75;A3-L-B76;A3-L-B77; A4~A548-L-B1;A4~A548-L-B2;A4~A548-L-B3;A4~A548-L-B4~B75;A4~A548-L-B76;A4~A548-L-B77; A549-L-B1;A549-L-B2;A549-L-B3;A549-L-B4~B75;A549-L-B76;A549-L-B77; A550-L-B1;A550-L-B2;A550-L-B3;A550-L-B4~B75;A550-L-B76;A550-L-B77.

[0097] Exemplary compounds, as defined herein, include each single compound listed in Tables 3 and 4 below, from Example 1 through Example 159.

[0098] Examples of preferred compounds are namely Examples 2, 4, 6, 7, 14, 16, 18, 30, 31, 33-37, 40, 43-46, 49, 51-60, 81, 84-88, 90, 93-99, 102-105, 108, 111, 112, 116-119, 122, 126, 127, 130, 131, 135-137, 139, 141, 144, 147, 148, 149, 153 and 158, or salts and / or solvates thereof.

[0099] Examples of more preferred compounds are Examples 4, 6, 7, 14, 16, 18, 30, 33, 35, 36, 37, 40, 43-45, 49, 51, 56-58, 85, 88, 95, 98, 99, 103, 104, 105, 111, 112, 116, 122, 135 and 136, or salts and / or solvates thereof.

[0100] It is understood that any of the above compounds may be in any amorphous, crystalline, or polymorphic form, or a combination thereof, including any salt or solvate form. The compounds herein may be further modified by appending various functional groups via any synthetic means defined herein to enhance selective biological properties. Such modifications are known in the art and include those that increase biological penetration into a given biological system (e.g., blood, lymphatic system, central nervous system), increase oral availability, increase solubility to allow administration by injection, alter metabolism, and alter excretion rate.

[0101] These compounds can be prepared by conventional chemical syntheses, such as those described above and illustrated in the schemes and examples of this disclosure. As can be appreciated by those of skill in the art, additional methods of synthesizing compounds of the formulae herein will be apparent to those of ordinary skill in the art. Additionally, the various synthetic steps can be performed in an alternate sequence or order to provide the desired compounds.

[0102] iii. Methods, Uses, Formulations and Administration

[0103] As used herein, the term "effective amount" refers to an amount of a drug or pharmaceutical agent that elicits the biological or medical response of a tissue, system, animal, or human that is being sought, for example, by a researcher or clinician. Moreover, the term "therapeutically effective amount" refers to an amount that results in the treatment, cure, prevention, amelioration of a disease, disorder, or symptoms thereof, or a reduction in the rate of progression of a disease or disorder, compared to a corresponding subject who does not receive such amount. This term also includes within its scope an amount effective to enhance normal physiological function.

[0104] As used herein, the terms "treatment," "treat," and "treating," as used herein, refer to reversing a disease or disorder, or one or more symptoms thereof, alleviating one or more symptoms, delaying the onset of one or more symptoms, or inhibiting the progression of one or more symptoms. In some embodiments, treatment may be administered after one or more symptoms have developed. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and / or in light of genetic or other susceptibility factors). Treatment may also be continued after symptoms have resolved, e.g., to prevent or delay their recurrence.

[0105] In one embodiment, the disease or condition to be treated is a proliferative disease or disorder, or a kinase-mediated disease or disorder. More specifically, the disease or disorder to be treated includes a proliferative disease or disorder, a developmental abnormality caused by dysregulation of the RAS-ERK signaling cascade (RASopathy), an inflammatory disease, or an immune system disorder.

[0106] According to some examples, the proliferative disease or disorder to be treated is a neoplasm, inflammatory disease or condition, or developmental abnormality associated with constitutively activating mutations in the RAS and / or RAF genes (e.g., KRAS and / or ARAF, BRAF, or CRAF mutations). The disease or disorder may also be further associated with receptor tyrosine kinase mutations or amplifications (e.g., EGFR, HER2), or mutations in downstream regulators of the receptors RAS (e.g., SOS1 gain of function, NF1 loss of function). For example, the compounds defined herein may be used to treat RAF mutations (e.g., BRAF V600EThe compounds of the present invention are inhibitors of signaling enzymes (e.g., BRAF and CRAF) that are involved in controlling cell proliferation not only in tumors harboring BRAF (Braf et al., 2002), but also, importantly, in the context of mutated RAS-driven cancers. Thus, the compounds of the present invention can be used, for example, for the treatment of diseases linked to the activity of these signaling enzymes and characterized by excessive or abnormal cell proliferation.

[0107] According to one embodiment, the disease or disorder is characterized by uncontrolled cell proliferation, i.e., a "proliferative disorder" or "proliferative disease." More specifically, these diseases and disorders relate to abnormal conditions characterized by autonomously proliferating cells, i.e., rapidly proliferating cell proliferation that generally forms distinct masses that exhibit a partial or total lack of structural organization and functional coordination with normal tissue.

[0108] For example, proliferative disorders or diseases are defined as "neoplasms," "neoplastic disorders," "neoplasia," "cancer," and "tumors," which are collectively meant to include hematopoietic neoplasms (e.g., lymphomas or leukemias) and solid neoplasms (e.g., sarcomas or carcinomas), which include, for example, precancerous and cancerous growths, or cancerous processes, metastatic tissues, or malignantly transformed cells, tissues, or organs of any kind, regardless of histopathological type or stage of invasiveness. Hematopoietic neoplasms are malignant tumors affecting hematopoietic structures (structures associated with the formation of blood cells) and components of the immune system, and include, for example, leukemias (associated with leukocytes (white blood cells) and their precursors in the blood and bone marrow) and lymphomas (associated with lymphocytes) arising from the myeloid, lymphoid, or erythroid lineages. Solid neoplasms include sarcomas, which are malignant neoplasms arising from connective tissues, such as muscle, cartilage, blood vessels, fibrous tissue, fat, or bone. Solid neoplasms also include carcinomas, which are malignant neoplasms arising from epithelial structures, including external epithelia (e.g., skin and the lining of the digestive tract, lungs, and cervix), and internal epithelia covering various glands (e.g., breast, pancreas, thyroid). Examples of neoplasms include leukemia, hepatocellular carcinoma, sarcoma, hemangioendothelial carcinoma, breast cancer, central nervous system cancers (e.g., astrocytoma, gliosarcoma, neuroblastoma, oligodendroglioma, and glioblastoma), prostate cancer, lung and bronchial cancer, laryngeal cancer, esophageal cancer, colon cancer, colorectal cancer, gastrointestinal cancer, melanoma, ovarian and endometrial cancer, kidney and bladder cancer, liver cancer, endocrine cancer (e.g., thyroid), and pancreatic cancer. For example, the disease or disorder is selected from colon cancer, lung cancer, pancreatic cancer, thyroid cancer, breast cancer, and skin cancer. Examples of neoplasms include melanoma, papillary thyroid carcinoma, colorectal cancer, ovarian cancer, breast cancer, endometrial cancer, liver cancer, sarcoma, gastric cancer, Barrett's adenocarcinoma, glioma (including ependymoma), lung cancer (including non-small cell lung carcinoma), head and neck cancer, acute lymphocytic leukemia, acute myeloid leukemia, non-Hodgkin's lymphoma, and hairy cell leukemia.

[0109] In one embodiment, a patient presenting with one of the above-described hematopoietic or solid neoplasms has previously been treated with, but has acquired resistance to, an inhibitor targeting the RAS-ERK pathway (including an RTK, RAF, MEK, or ERK inhibitor), including standard of care such as vemurafenib, dabrafenib, cobimetinib, trametinib, YERVOY, OPDIVO, or a combination of these agents.

[0110] In one embodiment, the disease to be treated is defined by a developmental abnormality caused by dysregulation of the RAS-ERK signaling cascade (a rasopathy: e.g., Noonan syndrome, Costello syndrome, LEOPARD syndrome, cardiac anomaly syndrome, and hypertrophic cardiomyopathy).

[0111] In one embodiment, the disease to be treated is defined as an inflammatory disease or immune system disorder, examples of which include inflammatory bowel disease, Crohn's disease, ulcerative colitis, systemic lupus erythematosis (SLE), rheumatoid arthritis, multiple sclerosis, thyroiditis, type 1 diabetes, sarcoidosis, psoriasis, allergic rhinitis, asthma, and chronic obstructive pulmonary disease (COPD).

[0112] In one embodiment, the compounds defined herein inhibit RAS-ERK signaling and cell proliferation in tumor cells harboring at least one mutated RAS or RAF genotype without inducing or substantially inducing paradoxical pathways.

[0113] As used herein, the term "patient or subject" refers to an animal, e.g., a mammal. Thus, a subject can refer to, for example, a mouse, rat, dog, cat, horse, cow, pig, guinea pig, a primate, such as those described above, including a human, and the like. Preferably, the subject is a human.

[0114] Thus, the present specification further relates to a method of treating a subject, e.g., a human subject, e.g., a subject suffering from a proliferative disease or disorder, e.g., a subject suffering from a RAF-mutated cancer and / or a mutated RAS-driven cancer, comprising administering to a subject in need of such treatment a therapeutically effective amount of a compound as defined herein.

[0115] In some embodiments, the present disclosure provides a method for treating a disorder (described herein) in a subject, comprising administering a compound of the present disclosure to a subject identified as needing such treatment. Identifying patients in need of treatment for the aforementioned disorders is within the ability and knowledge of those skilled in the art. Certain methods for identifying patients at risk of developing the aforementioned disorders that can be treated by the subject methods are understood in the medical arts, such as family history and the presence of risk factors associated with the development of the disease state in the subject patient. Those skilled in the medical arts can easily identify such candidate patients using, for example, clinical tests, physical examinations, medical / family history, and genetic assessments.

[0116] A method for assessing the effectiveness of a treatment in a subject includes determining pre-treatment symptoms of the disorder by methods known in the art, and then administering a therapeutically effective amount of a compound of the present invention to the subject. After an appropriate period (e.g., 1 week, 2 weeks, 1 month, 6 months) after administration of the compound, the symptoms of the disorder are again determined. Modulation (e.g., a decrease) of symptoms and / or biomarkers of the disorder (e.g., pERK or pMEK) indicates the effectiveness of the treatment. The symptoms and / or biomarkers of the disorder can be determined periodically throughout the treatment period. For example, the symptoms and / or biomarkers of the disorder can be checked every few days, weeks, or months to further evaluate the effectiveness of the treatment. A decrease in the symptoms and / or biomarkers of the disorder indicates that the treatment is effective.

[0117] In some embodiments, a therapeutically effective amount of a compound defined herein can be administered to a patient alone or in a composition in admixture with a pharmaceutically acceptable carrier, adjuvant, or vehicle.

[0118] The expression "pharmaceutically acceptable carrier, adjuvant, or vehicle" and equivalent expressions refer to a non-toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound it is formulated in. Pharmaceutically acceptable carriers, adjuvants, or vehicles that can be used in the compositions of the present disclosure include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and wool fat.

[0119] The compositions described herein can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, or via an implanted reservoir. As used herein, the term "parenteral" includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intracavity, and intracranial injection or infusion techniques. Other modes of administration also include intradermal or transdermal administration.

[0120] Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compound, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, and mixtures thereof. In addition to inert diluents, oral compositions may also include adjuvants, such as wetting agents, emulsifiers and suspending agents, surfactants, sweeteners, flavorings, and fragrances.

[0121] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions, can be formulated according to known techniques using suitable dispersing or wetting agents and suspending agents. Sterile injectable preparations can also be sterile injectable solutions, suspensions, or emulsions in non-toxic parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol. Among the acceptable vehicles and solvents that can be used are water, Ringer's solution, USP, and isotonic sodium chloride solution. In addition, sterile fixed oils are commonly used as solvents or suspending media. For this purpose, any brand of fixed oil, including the above-mentioned fixed oils, including synthetic monoglycerides or diglycerides, can be used. In addition, fatty acids, such as oleic acid, are used in the preparation of injectables.

[0122] Injectable formulations can be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium before use.

[0123] To prolong the effect of a given compound, it is often desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This can be accomplished by using a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the compound then depends on its rate of dissolution, which in turn depends on crystal size and crystalline form. Alternatively, delayed absorption of parenterally administered compounds can be achieved by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are prepared by forming microencapsule matrices of the compound in biodegradable polymers, such as polylactide-polyglycolide. The release rate of the compound can be controlled by the ratio of the compound to the polymer and the nature of the polymer used.

[0124] Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes and microemulsions which are compatible with body tissue.

[0125] Compositions for rectal administration are preferably suppositories, which can be prepared by mixing the compounds herein with a suitable non-irritating excipient or carrier, such as cocoa butter, polyethylene glycol or a suppository wax, which is solid at ambient temperature but liquid at body temperature and therefore melts in the rectum and releases the active compound.

[0126] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate, and / or a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; b) binders, such as carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidinone (PVP), sucrose, and acacia; c) humectants, such as glycerol; d) disintegrants, such as agar ( agar-agar), calcium carbonate, potato starch or tapioca starch, alginic acid, certain silicates and sodium carbonate, e) solution retardants such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as cetyl alcohol and glyceryl monostearate, h) adsorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also contain buffering agents.

[0127] Solid compositions of a similar type can also be used as fillers for soft and hard-filled gelatin capsules using additives such as lactose or milk sugar and high molecular weight polyethylene glycols. Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical formulation art. They may optionally contain opacifying agents and can also be composed so that they release one or more active ingredients only, or preferentially, in a certain part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type can also be used as fillers for soft and hard-filled gelatin capsules using additives such as lactose or milk sugar and high molecular weight polyethylene glycols.

[0128] The compositions may also be in microencapsulated form with one or more additives, as described above. Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings, release-controlling coatings, and other coatings well known in the pharmaceutical formulation art. In such solid dosage forms, the active compound may be admixed with at least one inert diluent, such as sucrose, lactose, or starch. Such dosage forms may also contain, as is common practice, additional substances other than inert diluents, such as tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pills, the dosage forms may also contain buffering agents. They may optionally contain opacifying agents, and may be of a composition that releases one or more active ingredients only, or preferentially, in a certain part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

[0129] Dosage forms for topical or transdermal administration of the compounds herein include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches. The active ingredient is admixed under sterile conditions with a pharmaceutically acceptable carrier and any required preservatives or buffers, as needed. Ophthalmic formulations, ear drops, and eye drops are also contemplated herein. Additionally, the present invention contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of the compound into the body. Such dosage forms can be prepared by dissolving or dispensing the compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0130] The pharmaceutically acceptable compositions provided herein may also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the art of pharmaceutical formulation and may be prepared as solutions in saline using benzyl alcohol or other suitable preservatives, absorption enhancers to enhance bioavailability, fluorocarbons, and / or other conventional solubilizing or dispersing agents.

[0131] The pharmaceutically acceptable compositions provided herein can be formulated for oral administration. Such formulations can be administered with or without food. In some embodiments, the pharmaceutically acceptable compositions of the present disclosure are administered without food. In other embodiments, the pharmaceutically acceptable compositions of the present disclosure are administered with food.

[0132] The amount of compound that can be combined with a carrier material to produce a composition in a single dosage form will vary depending upon the patient to be treated and the particular mode of administration. Provided compositions can be formulated so that a dosage of 0.01 to 100 mg / kg body weight / day of inhibitor can be administered to a patient receiving these compositions.

[0133] It should also be understood that the specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, the judgment of the treating physician, and the severity of symptoms associated with the proliferative disease or disorder. The amount of compound provided in the composition will also depend on the particular compound in the composition.

[0134] The compounds or compositions described herein can be administered in any amount and by any route of administration effective for treating or reducing the severity of symptoms as contemplated herein. The exact amount required will vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the infection, the specific drug, its mode of administration, and the like. The provided compounds are preferably formulated into unit dosage forms for ease of administration and uniformity of dosage. As used herein, the expression "unit dosage form" refers to a physically discrete unit of drug appropriate for the patient to be treated. However, it will be understood that the total daily usage of the compounds and compositions of the present disclosure will be determined by the attending physician within the scope of sound medical judgment.

[0135] The pharmaceutically acceptable compositions of the present disclosure can be administered to humans and other animals orally, rectally, parenterally, intraperitoneally, topically (by powder, ointment, or drops), bucally, as an oral or nasal spray, etc., depending on the severity of the infection being treated. In some embodiments, provided compounds can be administered orally or parenterally at a dosage of about 0.01 mg / kg to about 50 mg / kg, preferably about 1 mg / kg to about 25 mg / kg, of the subject's body weight per day to achieve the desired therapeutic effect, one or more times daily.

[0136] It will be understood that the total daily usage of the compounds and compositions herein can be determined by the attending physician within the scope of sound medical judgment. The total daily inhibitory dose of the compounds herein, administered to a subject in single or divided doses, can be, for example, in an amount of 0.01 to 50 mg / kg body weight, more usually 0.1 to 25 mg / kg body weight. Single dose compositions can contain such amounts or multiples thereof to make up the daily dosage. In one embodiment, a treatment regimen in accordance with the present invention comprises administering to a patient in need of such treatment, in single or multiple doses, from about 10 mg to about 1000 mg of one or more compounds herein per day.

[0137] Depending on the disease or disorder to be treated, additional therapeutic agents may also be present in the compositions of the present disclosure or may be administered separately as part of a dosing regimen, e.g., an additional chemotherapeutic agent. Non-limiting examples of additional therapeutic agents that can be used in combination with the compounds of the invention include antiproliferative compounds, such as aromatase inhibitors; antiestrogens; antiandrogens; gonadorelin agonists; topoisomerase I inhibitors; topoisomerase II inhibitors; microtubule activators; alkylating agents; retinoids, carotenoids, tocopherols; cyclooxygenase inhibitors; MMP inhibitors; antimetabolites; platin compounds; methionine aminopeptidase inhibitors; bisphosphonates; antiproliferative antibodies; heparanase inhibitors; inhibitors of Ras oncogenic isoforms; telomerase inhibitors; proteasome inhibitors; compounds used in the treatment of hematological malignancies; kinesin spindle protein inhibitors; Hsp90 inhibitors; mTOR inhibitors; PI3K inhibitors; Flt-3 inhibitors; CDK4 / 6 inhibitors; HER2 inhalants (Herceptin, EGFR inhibitors (Iressa, Tarceva, Nerlynx, Tykerb, Erbitux); RAS inhibitors; MEK inhibitors (Trametinib, Binimetinib, Cobimetinib); ERK inhibitors (Ulixertinib); anti-PD-1 antibodies (Opdivo), (Keytruda); anti-CTLA4 antibodies (Yervoy); antitumor antibiotics; nitrosoureas; compounds that target / increase protein or lipid kinase activity, or compounds that target / increase protein or lipid phosphatase activity, or further anti-angiogenic compounds.

[0138] The treatment may also be supplemented with other treatments or interventions, such as surgery, radiation therapy (e.g., gamma irradiation, neutron irradiation, electron beam irradiation, proton therapy, brachytherapy, and systemic radioisotopes), biologic response modifiers (e.g., interferons, interleukins, tumor necrosis factors (TNF)), and medications used to reduce side effects.

[0139] The description of an embodiment of a variation herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof. The description of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof.

[0140] Example

[0141] List of abbreviations: Ac: Acetyl AcOEt or EtOAc: ethyl acetate AcOH: acetic acid Ar: aryl ATCC: American Type Culture Collection ATP: adenosine triphosphate BINOL: [1,1'-binaphthalene]-2,2'-diol Boc: tert-butyloxycarbonyl BOP: (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate br: Broad BSA: bovine serum albumin CCL: cancer cell lines CDCl3: deuterated chloroform DCE: 1,2-dichloroethane DCM: dichloromethane DIEA (or DIPEA): N,N-diisopropylethylamine (Huenig's base) DME: 1,2-dimethoxyethane DMF: N,N-dimethylformamide DMSO: dimethyl sulfoxide DMSO-d6: Deuterated dimethyl sulfoxide DTT: dithiothreitol EA: Ethyl acetate EC 50 : Half-maximal effective concentration ECL: enhanced chemiluminescence EDTA: Ethylenediaminetetraacetic acid Et2O: Diethyl ether EtOH: ethanol Eu: Europium FBS: fetal bovine serum GST: glutathione S-transferase HATU: O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate HEPES: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid Het: Heterocycle Hex: Hexane HRMS: high resolution mass spectrometry HPLC: high performance liquid chromatography HRP: horseradish peroxidase I C 50 :half-maximal inhibitory concentration IPA: Isopropanol iPrOH: Isopropanol LCMS: liquid chromatography mass spectrometry MeCN: acetonitrile MS: mass spectrometry NMP: N-methylpyrrolidone NMR: nuclear magnetic resonance ON: overnight PBS: phosphate buffered saline pERK: phosphorylated extracellular signal-regulated kinase PMB: para-methoxybenzyl PMSF: Phenylmethylsulfonyl fluoride Rf: retention factor RPMI-1640: Roswell Park Memorial Institute medium RT or Rt: room temperature SDS: sodium dodecyl sulfate SDS-PAGE: sodium dodecyl sulfate-polyacrylamide gel electrophoresis SEM: Trimethylsilylethoxymethyl SNAr: Nucleophilic aromatic substitution TBST: 0.2% Tween (登録商標) Tris-buffered saline containing -20 TBTU: O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium trifluoroborate TEV: tobacco etch virus protease TFA: Trifluoroacetic acid THF: tetrahydrofuran TLC: Silica gel thin layer chromatography Ts: para-toluenesulfonate Y 最小 :Minimal data point of a dosage-activity curve

[0142] The following non-limiting examples are illustrative embodiments and should not be construed as further limiting the scope of the present invention. These examples will be better understood with reference to the accompanying figures.

[0143] The examples set forth herein below provide synthesis and experimental results obtained for certain exemplary compounds. As is well known to those skilled in the art, reactions are carried out in an inert atmosphere (nitrogen or argon) as necessary to protect reaction components from air and moisture. Temperatures are given in degrees Celsius (°C). Solution percentages and ratios are expressed volume-to-volume unless otherwise noted. Reactants used in the following examples may be obtained as described herein or, if not described herein, are themselves commercially available or may be prepared from commercially available materials by methods known in the art. Flash chromatography is performed on silica (SiO2) at 254 nm using commercially available normal-phase silica on a Teledyne Isco Rf Combiflash instrument. Mass spectrometry is recorded using an electrospray mass spectrometer. NMR was recorded on a 400 MHz Varian instrument or a 600 MHz Bruker instrument.

[0144] Preparative HPLC was performed using Agilent equipment with a Phenomenex-Kinetex C18 (21 x 100 mm, 5 μm) column at a flow rate of 20 mL / min (RT) and UV detection at 220 nm and 254 nm. The mobile phase consisted of solvent A (5% MeOH, 95% water + 0.1% formic acid) and solvent B (95% MeOH, 5% water + 0.1% formic acid) unless otherwise noted. Where specified herein, 0.05% TFA or 0.1% AcOH or other additives were sometimes used in place of 0.1% formic acid in both solvents. As specified herein, for more difficult separations, MeCN was used in place of MeOH in both mobile phases. Specific gradient conditions are provided in the Examples below, but the following are representative: T(0) → T(3 min) was run isocratically with 10-50% solvent B depending on the polarity of the compound, followed by a 12 min gradient to 100% solvent B, with the final 5 min at 100% solvent B.

[0145] LCMS analysis was performed on an Agilent instrument. Liquid chromatography was performed on a Phenomenex Kinetex C18 column (2.6 μm; 100 Å; 3 × 30 mm) at a flow rate of 1.5 mL / min (RT) with UV detection at 220 and 254 nm. The mobile phase consisted of solvent A (95% HO / 5% MeOH / 0.1% AcOH) and solvent B (95% MeOH / 5% HO / 0.1% AcOH) with the following gradient: T(0) 100% A → T(0.5 min) 100% B → isocratic 100% B → T(2 min). MS detection was performed in parallel using APCI detection in both positive and negative modes.

[0146] Unless otherwise indicated, all numerical values ​​expressing amounts of ingredients, reaction conditions, concentrations, properties, stability, and the like used in the specification and appended claims are understood to be modified in all instances by the word "about." At the very least, each numerical parameter should be construed in light of, at least, the number of reported significant digits and by applying ordinary rounding techniques. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and appended claims are approximations that may vary depending upon the properties sought to be obtained. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the embodiments are approximations, the numerical values ​​set forth in the specific examples are reported as precisely as possible. However, any numerical value inherently contains certain errors resulting from variations in experiments, testing measurements, statistical analyses, and the like.

[0147] Synthesis, biological activity and properties of examples:

[0148] All compounds defined herein were prepared according to the methods shown in Tables 3 and 4. Mass spectrometry and NMR characterization data are provided for each example. Compounds were tested in the assays described in the Biological Experiments section. Conventions used for reporting biological data are provided as footnotes to each table.

[0149] Synthesis method A:

[0150] Commercially available 2,6-difluoroaniline A-1 is converted to its acetanilide A-2 using an acetylating agent such as acetic anhydride, as described in International Publication WO 2012 / 101238 A1, and then converted to the monoprotected dianiline A-3. Sulfonylation to sulfonamide A-4 can be achieved using a sulfonylating reagent such as a sulfonyl chloride in the presence of an organic base such as pyridine, with or without a catalyst such as 4-dimethylaminopyridine, and a solvent such as dichloromethane or tetrahydrofuran. Treatment of acetanilide A-4 with aqueous hydrochloric acid in the presence of a cosolvent such as an alcohol provides the aniline salt A-5.

[0151] Commercially available 2-amino-2-cyanoacetamide A-6 can be converted to 5-amino-2-(methylthio)thiazole-4-carboxamide A-8 according to a two-step procedure described in Heterocycl. Commun., 2014, 20, 175. Intermediate A-8 can then be cyclized to the corresponding pyrimidone by heating in formamide, as described in Indian J. Chem., 2010, 49B, 1229. Subsequent treatment with a chlorinating agent, such as thionyl chloride or phosphoryl chloride, in the presence of a catalytic amount of DMF, according to a procedure also described in Indian J. Chem., 2010, 49B, 1229, affords chloro-compound A-10.

[0152] Intermediate A-11 can be obtained by heating chloropyrimidine A-10 with aniline salt A-5 in an organic acid, such as acetic acid. Inhibitors of general formula WI are prepared from intermediates of general formula A-11 by a two-step procedure: first, the thiomethyl group is oxidized, typically to the corresponding methyl sulfoxide and methyl sulfone mixture, which is then reacted with a nucleophile (e.g., a primary or secondary amine, or an NH-containing heterocycle). The latter step is typically carried out in a solvent such as DMSO or NMP in the presence of a base (e.g., an organic base such as DIEA, trimethylamine, pyridine, or an inorganic base such as potassium carbonate, cesium carbonate, and sodium carbonate) at a temperature ranging from 70 to 140°C.

[0153] [ka]

[0154] Synthesis method B:

[0155] Inhibitors of general formula W-II are prepared as described in synthetic method B. Intermediates of general formula A-11 are first prepared according to synthetic method A and then oxidized to a mixture of methyl sulfoxide and methyl sulfone as described in synthetic method A. These are then coupled to 3-indole carboxylic esters (e.g., methyl esters, X = CH) or 3-indazole carboxylic esters (e.g., methyl esters, X = N) according to a protocol similar to that described for the introduction of nucleophiles in synthetic method A. The latter step is typically carried out in the presence of a base (e.g., an organic or inorganic base, such as CsCO, KOtBu, DIEA, trimethylamine, pyridine, etc.) in a solvent such as THF, DMSO, or NMP at temperatures ranging from 60 °C to 140 °C. Deprotection of the ester protecting group using an inorganic base (e.g., NaOH or KOH) in a mixture of water and a miscible organic solvent (e.g., methanol, ethanol, THF, dioxane, etc.) at temperatures ranging from ambient to 100 °C, followed by acidification with a mineral (e.g., aqueous hydrochloric acid or sulfuric acid), an inorganic salt solution (e.g., aqueous NHCl or KHSO), or an organic acid (e.g., aqueous citric acid or acetic acid) provided the corresponding carboxylic acid intermediate B-1 or B-2. Coupling of intermediate B-1 or B-2 with an amine using a standard amide coupling reagent (e.g., TBTU, HATU, DCC, EDC, etc.) provided the amide derivatives of general formula W-II.

[0156] [ka]

[0157] Synthesis method C:

[0158] Cross-coupling of commercially available bromobenzimidazole C-1 to heteroaryl boronic acids or boronic esters can be carried out under palladium-catalyzed Suzuki-Miyaura cross-coupling conditions in a solvent such as dioxane or dimethoxyethane in the presence of a base such as sodium carbonate or potassium carbonate to give intermediate C-2. The substituted benzimidazole derivative C-2 can then be coupled to intermediate A-11 after oxidation of the methyl sulfide moiety as described in the general protocol of Method A to provide inhibitors of general structure W-III.

[0159] [ka]

[0160] Synthesis method D:

[0161] Deprotected 3-indole sulfonyl chloride D-2 can be prepared as described in Org. Lett. 2011, 13, 3588. This reagent can be converted to the corresponding sulfonamide D-3 by reaction with a primary or secondary amine in the presence of an organic base, such as DIEA or triethylamine. The D-3 fragment can then be coupled to intermediate A-11 after oxidation of the methyl sulfide moiety, as described in the general protocol of Method A, to provide inhibitors of general structure W-IV.

[0162] [ka]

[0163] Synthesis method E:

[0164] Alternatively, inhibitors of general structure W-IV can be obtained via N-tosyl-protected indole-3-sulfonyl chloride E-1, prepared according to the procedure described in Chemical and Pharmaceutical Bulletin 2009, 57, 591. E-1 can then be converted to the corresponding sulfonamide E-2 by reaction with a primary or secondary amine in the presence of an organic base, such as DIEA or triethylamine, followed by removal of the tosyl protecting group with an aqueous inorganic base, such as KOH. The E-2 fragment can then be coupled to intermediate A-11 after oxidation of the methyl sulfide moiety as previously described.

[0165] [ka]

[0166] Synthesis method F:

[0167] 3-Indole thiocyanate F-1 (prepared according to the procedure described in Phosphorus, Sulfur, and Silicon and the Related Elements 2014, 189, 1378) is reduced to the corresponding sulfide salt using a reducing agent, such as sodium sulfide anhydrate, and directly alkylated with an alkyl halide without isolation to provide sulfide intermediate F-2. Sulfide intermediate F-2 is then converted to sulfone intermediate F-3 using an oxidizing agent, such as 3-chloroperoxybenzoic acid. The final inhibitor of general structure WV is then obtained by coupling indole sulfone F-3 with the A-11 intermediate after oxidation of the methyl sulfide moiety as previously described.

[0168] [ka]

[0169] Synthesis method G:

[0170] A bright red solution of commercially available 3-fluoro-2-nitroaniline G-1 can be reacted with a primary or secondary amine in a solvent such as MeCN, DMSO, NMP, or DMF in the presence of an inorganic base such as potassium carbonate or an organic base such as DIEA, and heated at temperatures ranging from 40°C to 120°C under thermal or microwave conditions to give intermediate G-2. Reduction of the nitro group in intermediate G-2 and subsequent cyclization of the resulting 1,2-phenylenediamine intermediate to the desired benzimidazole intermediate G-3 can be carried out in a single operation using metallic iron, ammonium chloride, and formic acid in an alcoholic solvent such as isopropanol, and heated at temperatures ranging from 40°C to 90°C. The final inhibitors of general structure W-VI can then be obtained by heating intermediate A-11 to the corresponding substituted benzimidazole G-3, which has been oxidized to its corresponding methyl sulfone / methyl sulfoxide, in the presence of a base under standard conditions as previously described.

[0171] [ka]

[0172] Procedures for preparing inhibitors that do not fall within or only partially fall within the general synthetic procedures described above are specifically described below.

[0173] Sulfonyl chlorides:

[0174] The following sulfonyl chlorides were obtained from commercial sources and used as received: 2-chlorobenzenesulfonyl chloride, 2,3-dichlorobenzenesulfonyl chloride, 3-chloro-2-methylbenzenesulfonyl chloride, 3-fluoro-2-methylbenzenesulfonyl chloride.

[0175] Other sulfonyl chlorides were prepared by using or adapting literature procedures, as described below.

[0176] 3-Fluoro-2-methyl-4-methoxybenzenesulfonyl chloride: [ka]

[0177] Step 1: To a solution of 2-fluoro-3-methylphenol (4.32 mL, 39.7 mmol) in acetone (50 mL) was added potassium carbonate (6.58 g, 47.6 mmol), followed by iodomethane (2.75 mL, 43.7 mmol). The reaction mixture was then refluxed at 60 °C overnight. The reaction mixture was then cooled to room temperature, filtered (washed with 2 x 10 mL of acetone), and concentrated under reduced pressure. The crude product was extracted with water (30 mL) and EtOAc (2 x 50 mL). The organic layer was then separated, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel using 0-5% EtOAc / hexane to give 2-fluoro-3-methylanisole as a clear, colorless liquid (5.30 g, 95% yield): 1 H NMR(CDCl3) δ:6.95(td,J=8.0,1.4Hz,1H),6.85~6.71(m,2H),3.87(s,3H),2.28(d,J=2.3Hz,3H).

[0178] Step 2: To a solution of 2-fluoro-3-methylanisole (1.00 g, 7.13 mmol) from Step 1 in DCM (5.6 mL) was added a solution of chlorosulfonic acid (1.13 mL, 16.5 mmol) in DCM (5.6 mL) over 5 min. The pale brown reaction mixture containing a viscous liquid layer was stirred at room temperature for 10 min and then quenched by pouring into a mixture of water (10 mL) and ice (5 g). The aqueous phase was extracted with DCM (2 x 10 mL), dried over NaSO, filtered, and concentrated under reduced pressure to give the desired sulfonyl chloride (1.70 g, 100% yield) as a colorless liquid. This material was used without further purification: 1H NMR(CDCl3) δ:7.87(dd,J=9.0,1.8Hz,1H),6.97~6.86(m,1H),3.97(s,3H),2.66(d,J=2.8Hz,3H).

[0179] 3-chloro-2-methyl-4-methoxybenzenesulfonyl chloride: [ka]

[0180] Following the same procedure as for 3-fluoro-2-methyl-4-methoxybenzenesulfonyl chloride (step 1), starting from 2-chloro-3-methylphenol, 2-chloro-3-methylanisole was obtained as a colorless liquid in quantitative yield: 1 H NMR(CDCl3) δ:7.12(t,J=7.9Hz,1H),6.87~6.83(m,1H),6.79(d,J=8.2Hz,1H),3.89(s,3H),2.38(s,3H).

[0181] Treatment with chlorosulfonic acid as described for 3-fluoro-2-methyl-4-methoxybenzenesulfonyl chloride (step 2) gave the desired 3-chloro-2-methyl-4-methoxybenzenesulfonyl chloride as a colorless liquid in 96% yield: 1 H NMR(CDCl3) δ:8.02(d,J=9.1Hz,1H),6.90(d,J=9.1Hz,1H),4.00(s,3H),2.83(s,3H).

[0182] General Synthetic Method A: Preparation of Difluoroaniline Hydrochloride Intermediate A-5 (Ar=2,3-Dichlorophenyl) from Acetanilide A-3: [ka]

[0183] Preparation of acetanilide A-2: Acetanilide A-2 can be prepared by acetylating 2,6-difluoroaniline A-1 with acetic anhydride according to the literature procedure described in Bioorg. Med. Chem. 2016, 24, 2215. Intermediate A-2 is converted to acetanilide A-3 by sequential nitration followed by reduction of the nitro group to the aniline as described in International Publication No. WO 2012 / 101238 A1.

[0184] Step 1: Preparation of sulfonamide A-4 (Ar = 2,3-dichlorophenyl): Aniline A-3 (8.50 g, 45.5 mmol) was dissolved in THF (145 mL), and pyridine (4 equiv., 14.7 mL) was added to the brown solution, followed by 2,3-dichlorobenzenesulfonyl chloride (1.2 equiv., 13.45 g). The resulting reaction mixture was stirred at 45 °C for 3.5 h, after which the conversion was judged complete as monitored by LCMS. The reaction mixture was cooled to room temperature and then partitioned between EtOAc and 2-Me-THF (1:1) and water. 1 N HCl solution was added until the pH was slightly acidic. A significant amount of off-white solid was present in the biphasic mixture and filtered off (first crop). The layers of the filtrate were separated, and the aqueous layer was extracted twice more with EtOAc. The combined organic extracts were washed once with water, then once with brine, dried over MgSO4, filtered, and concentrated to approximately 20 mL. The resulting suspension was sonicated, and the solid was collected by filtration and washed with EtOH (second crop). Both crops were combined and dried under reduced pressure. A-4 (15.3 g, 85% yield) was obtained as a beige solid, which was used without further purification: 1H NMR(DMSO-d6) δ:10.61(s,1H),9.67(s,1H),7.95(dd,J=8.0,1.4Hz,1H),7.85(dd,J=8.0,1.4Hz,1H),7.51(t,J=8.0Hz,1H),7.05~7.18(m,2H),2.00(s,3H). MS m / z 395.0(MH + ).

[0185] Step 2: Preparation of aniline hydrochloride A-5 (Ar = 2,3-dichlorophenyl): In a 500 mL round-bottom flask, acetanilide A-4 (7.00 g, 17.7 mmol) was suspended in ethanol (65 mL), and a 1:1 mixture of concentrated HCl and water (65 mL) was added. The flask was fitted with a stoppered reflux condenser and heated to 80 °C with stirring. After 24 h, the conversion was approximately 70%, as determined by LCMS monitoring. Additional EtOH (65 mL) and 6 N HCl (65 mL) were added to the suspension, and stirring was resumed at 80 °C for an additional 7 h, after which LCMS indicated complete conversion to the desired aniline. The reaction mixture was diluted with 50 mL of water while warm and filtered through a cotton plug to remove a small amount of insoluble material. It was then concentrated to dryness under reduced pressure. The residue was azeotropically dried by evaporating toluene under reduced pressure three times and then dried under vacuum to give 7.2 g of the desired product A-5 as its HCl salt in the form of a yellow solid: 1 H NMR(DMSO-d6) δ:10.30(s,1H),7.93(dd,J=8.2,1.2Hz,1H),7.83(dd,J=8.0,1.4Hz,1H),7.49(t,J=8.0Hz,1H),6.68~6.96(m,1H),6.31(td,J=8.6,5.5Hz,1H). MS m / z 350.9(MH).

[0186] The following A-5 intermediates (Table 1) were prepared in a similar manner using the relevant sulfonyl chlorides.

[0187] [Table 1]

[0188] General Synthetic Method A - Preparation of Inhibitor WI from Intermediate A-5: [ka]

[0189] Steps 1 and 2: Preparation of intermediate A-8: These two steps were carried out as described by Wang et al. in Heterocycl. Commun., 2014, 20, 175.

[0190] Step 3: Preparation of Intermediate A-9 (adapted from Indian J. Chem., 2010, 49B, 1229): 5-Amino-2-methylsulfanyl-thiazole-4-carboxamide (3.19 g, 16.855 mmol) was equally divided into two 20 mL microwave-safe vials, and formamide (13.4 mL, 337 mmol) was added to each. The vials were sealed and heated in a microwave oven at 185 °C for 20 minutes, then at 190 °C for an additional 15 minutes. The mixtures were cooled to room temperature, then combined and slowly added to ice-cold water (60 mL) containing 1.3 mL of acetic acid. The resulting yellow solid was collected by filtration and washed with several portions of cold water. After suction drying on the filter, the solid was transferred to a vial and dried under reduced pressure. This gave 2-methylsulfanyl-6,7a-dihydro-3aH-thiazolo[5,4-d]pyrimidin-7-one (2.34 g, 69% yield) as a yellow to beige solid. MS m / z 202.2 (MH + ). 1 H NMR (DMSO-d6) δ: 12.81 (br.s., 1H), 8.15 (s, 1H), 2.75 (s, 3H).

[0191] Step 4: Preparation of intermediate A-10 (adapted from Indian J. Chem., 2010, 49B, 1229): 2-Methylsulfanyl-6H-thiazolo[5,4-d]pyrimidin-7-one (1.00 g, 5.02 mmol) was suspended in thionyl chloride (10 mL, 137 mmol), and then 3 drops of DMF were added. The resulting mixture was refluxed in an oil bath set at 90 °C. After stirring for 3 hours, the mixture became a clear solution. After cooling to room temperature, it was then diluted with some toluene and concentrated to dryness under reduced pressure. The residue was suspended in toluene and once again concentrated to dryness. The product was then dried under reduced pressure. 7-Chloro-2-methylsulfanyl-thiazolo[5,4-d]pyrimidine (1.10 g, 100% yield) was obtained as a beige to brown solid, which was used without further purification. MS m / z 218.0 (MH + ). 1 H NMR (chloroform-d) δ: 8.77 (s, 1H), 2.88 (s, 3H).

[0192] Step 5: Preparation of Intermediate A-11 (Ar = 2,3-dichlorophenyl): N-(3-amino-2,4-difluoro-phenyl)-2,3-dichloro-benzenesulfonamide hydrochloride (811 mg, 2.08 mmol) and 7-chloro-2-methylsulfanyl-thiazolo[5,4-d]pyrimidine (544 mg, 2.50 mmol) were placed in a 20 mL vial and suspended in 10 mL of acetic acid. The mixture was then heated at 65 °C for 23 hours. It was then cooled to room temperature and slowly poured into 30 mL of water containing sodium acetate (370 mg, 4.51 mmol). The resulting suspension was sonicated to homogenize, and the solid was collected by filtration and washed with a small amount of water. The solid was sucked dry on the filter and further dried under reduced pressure. 2,3-Dichloro-N-[2,4-difluoro-3-[(2-methylsulfanylthiazolo[5,4-d]pyrimidin-7-yl)amino]phenyl]benzenesulfonamide (1.17 g, 100% yield) was obtained as a yellow solid, which was used without further purification. MS m / z 534.0 (MH + ). 1 H NMR(DMSO-d6) δ:10.65(s,1H),9.76(s,1H),8.26(s,1H),7.95(dd,J=8.0,1.4Hz,1H),7.88(dd,J=8.0,1.4Hz,1 H),7.52(t,J=8.0Hz,1H),7.24(td,J=8.6,5.9Hz,1H),7.17(td,J=9.4,1.2Hz,1H),2.79(s,3H).

[0193] The following A-11 intermediates were prepared in a similar sequence as described above (Table 2).

[0194] [Table 2]

[0195] Step 6: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 (Ar = 2,3-dichlorophenyl): 2,3-Dichloro-N-[2,4-difluoro-3-[(2-methylsulfanylthiazolo[5,4-d]pyrimidin-7-yl)amino]phenyl]benzenesulfonamide (1.00 g, 1.87 mmol) was suspended in DCM (25 mL). 3-Chloroperoxybenzoic acid (839 mg, 3.74 mmol) was then added in one portion. The mixture was stirred at room temperature for 23 h. A saturated solution of sodium thiosulfate was added (approximately 1 mL), and the mixture was stirred vigorously for 10 min. It was then diluted with DCM and washed three times with a 1:1 mixture of saturated NaHCO3 and water. The aqueous layer was neutralized (pH 7) by slowly adding acetic acid. The aqueous layer was then back-extracted twice with DCM, and the combined organic layers were washed once with water and then with brine. The organic layer was then dried over Na2SO4, filtered, concentrated, and dried under reduced pressure. The resulting beige solid was purified by filtration using Celite®. (登録商標) The compound was adsorbed onto silica gel and then purified by flash chromatography on a 25 g silica gel column using a gradient of 5% to 100% EtOAc in hexanes. The following two main peaks were separated:

[0196] Methyl sulfone: rf=0.72 @ 8:2 EtOAc / hexane, 710 mg (67% yield) as a beige solid. MS m / z 566.2 (MH + ). 1 H NMR(DMSO-d6) δ:10.68(s,1H),10.52(s,1H),8.49(s,1H),7.96(dd,J=8.2,1.6Hz,1H),7.88(dd,J=7.8,1.6Hz, 1H),7.53(t,J=8.0Hz,1H),7.29(td,J=8.6,5.5Hz,1H),7.21(td,J=9.0,1.2Hz,1H),3.58(s,3H)

[0197] Methyl sulfoxide: rf=0.31 @ 8:2 EtOAc / hexane, 229.1 mg (22% yield) as a beige solid, MS m / z 550.2 (MH + ). 1 H NMR(DMSO-d6) δ:10.67(s,1H),10.24(s,1H),8.41(s,1H),7.96(dd,J=8.2,1.6Hz,1H),7.88(dd,J=8.0,1.4Hz, 1H),7.53(t,J=8.0Hz,1H),7.27(td,J=8.6,5.9Hz,1H),7.19(td,J=9.0,0.8Hz,1H),3.11(s,3H)

[0198] NOTE: Chromatographic separation and purification of the two products is not necessary for the subsequent reaction (i.e., step 7 of General Synthetic Method A), which can be used as a crude mixture.

[0199] Step 7: Preparation of WI (Ar = 2,3-dichlorophenyl, Het = 1-benzimidazolyl) Example 4: 2,3-Dichloro-N-[2,4-difluoro-3-[(2-methylsulfonylthiazolo[5,4-d]pyrimidin-7-yl)amino]phenyl]benzenesulfonamide (50 mg, 0.0883 mmol) and 1-H-benzimidazole (26 mg, 0.221 mmol) were dissolved in 0.6 mL of NMP in a 1-dram vial. N,N-Diisopropylethylamine (0.077 mL, 0.441 mmol) was then added, and the mixture was heated with stirring at 105° C. for 22 hours. The reaction was cooled to room temperature and then quenched by the addition of a solution of formic acid (0.1 mL) in methanol (1 mL). The solution was then filtered and purified by reverse-phase preparative HPLC using a 40% to 90% MeOH gradient in water with 0.1% formic acid modifier. Appropriate fractions were pooled and concentrated. The residue was lyophilized from a mixture of water and MeCN to give N-[3-[[2-(benzimidazol-1-yl)thiazolo[5,4-d]pyrimidin-7-yl]amino]-2,4-difluoro-phenyl]-2,3-dichloro-benzenesulfonamide (Example 4: 22 mg, 40% yield) as a beige solid.

[0200] Other examples of inhibitors prepared in a similar manner are listed under Method A in Table 3.

[0201] General Method for the Synthesis of Inhibitors of Formula W-II (Method B, X=CH) - Synthesis of Example 28: [ka]

[0202] Step 1: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 (Ar = 2-methyl-3-fluorophenyl): N-[2,4-difluoro-3-[(2-methylsulfanylthiazolo[5,4-d]pyrimidin-7-yl)amino]phenyl]-3-fluoro-2-methyl-benzenesulfonamide (449 mg, 0.902 mmol) was suspended in DCM (10 mL). 3-Chloroperoxybenzoic acid (384 mg, 1.71 mmol) was then added in one portion. The mixture was stirred at room temperature for 23 hours. A saturated solution of sodium thiosulfate was added (approximately 0.2 mL), and the mixture was stirred vigorously for 10 minutes. It was then diluted with DCM and washed three times with a 1:1 mixture of saturated NaHCO3 solution and water. The combined aqueous layers were back-extracted once with DCM, and the combined organic layers were washed once with water and then with brine. The organic layer was then dried over NaSO, filtered, concentrated, and dried under reduced pressure. The resulting beige solid (445.5 mg, 93% yield, as a 2:1 mixture of sulfone:sulfoxide) was used directly without purification. MS m / z 530.2 and 514.2 (MH + ).

[0203] Steps 2 and 3: Preparation of carboxylic acid B-1: The crude mixture of sulfone and sulfoxide from above (300 mg, 0.567 mmol) and indole-3-carboxylic acid methyl ester (149 mg, 0.850 mmol) were placed in a 20 mL vial and suspended in NMP (3.6 mL). DIPEA (0.49 mL, 2.83 mmol) was then added to the orange solution, and the reaction mixture was stirred at 105° C. for 24 h.

[0204] The reaction mixture was cooled to room temperature, and 4N sodium hydroxide solution (0.85 mL, 3.40 mmol) was added. The resulting mixture was stirred at 50°C for an additional 1 hour. The reaction mixture was cooled to room temperature and then transferred into 100 mL of 1N HCl. After sonicating the resulting suspension, the solids were collected by filtration and washed with water. They were then dried under reduced pressure to give 1-[7-[2,6-difluoro-3-[(3-fluoro-2-methyl-phenyl)sulfonylamino]anilino]thiazolo[5,4-d]pyrimidin-2-yl]indole-3-carboxylic acid (292 mg, 84.3% yield) as a brown solid. The crude material was used for the next step without further purification. MS m / z 611.2 (MH + ).

[0205] Step 4: Preparation of W-II Example 28: 1-[7-[2,6-Difluoro-3-[(3-fluoro-2-methyl-phenyl)sulfonylamino]anilino]thiazolo[5,4-d]pyrimidin-2-yl]indole-3-carboxylic acid (35 mg, 0.0573 mmol) and HATU (44 mg, 0.115 mmol) were dissolved in NMP (1 mL) followed by DIPEA (0.060 mL, 0.344 mmol). The orange solution was stirred at room temperature for 2-3 minutes, and then N-methyl-2-(pyridin-2-yl)ethan-1-amine (0.016 mL, 0.115 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours. The reaction was then quenched by the addition of 0.5 mL of formic acid, diluted with a small amount of DMSO, and then purified by preparative reverse-phase HPLC to give Example 28 (9.8 mg, 23% yield) as a beige solid after lyophilization from a mixture of MeCN / water.

[0206] Other examples of inhibitors prepared in a similar manner are listed under Method B in Table 3.

[0207] General method for the synthesis of inhibitors of formula W-II (Method B, X=N) - Synthesis of Example 23: [ka]

[0208] Step 1: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 was carried out as described above in Example 28 (Step 1).

[0209] Steps 2 and 3: Preparation of Carboxylic Acid B-2: The crude mixture of sulfone and sulfoxide from above (300 mg, 0.567 mmol) and methyl indazolyl-3-carboxylate (150 mg, 0.850 mmol) were placed in a 20 mL vial and suspended in NMP (3.6 mL). DIPEA (0.49 mL, 2.83 mmol) was then added to the orange solution, and the reaction mixture was stirred at 105° C. for 24 hours. It was then cooled to room temperature, and 4 N sodium hydroxide in water (0.85 mL, 3.40 mmol) was added. The resulting mixture was stirred at 50° C. for an additional hour.

[0210] The reaction mixture was cooled to room temperature and then transferred to 100 mL of 1N HCl. After sonicating the resulting suspension, the solid was collected by filtration, washed with water, and dried under reduced pressure to give 1-[7-[2,6-difluoro-3-[(3-fluoro-2-methyl-phenyl)sulfonylamino]anilino]thiazolo[5,4-d]pyrimidin-2-yl]indazole-3-carboxylic acid (300 mg, 87% yield) as a light brown solid. MS m / z 612.2 (MH + ).

[0211] Step 4: Preparation of W-II Example 23: 1-[7-[2,6-Difluoro-3-[(3-fluoro-2-methyl-phenyl)sulfonylamino]anilino]thiazolo[5,4-d]pyrimidin-2-yl]indazole-3-carboxylic acid (35 mg, 0.0572 mmol) and HATU (44 mg, 0.114 mmol) were dissolved in NMP (1 mL). DIPEA (0.060 mL, 0.343 mmol) was then added. The orange solution was stirred at room temperature for 2-3 minutes, and then N-methyl-1-(3-pyridinyl)methanamine (0.013 mL, 0.114 mmol) was added.

[0212] The reaction mixture was stirred overnight at room temperature. The reaction was then quenched by the addition of 0.5 mL of formic acid, diluted with DMSO, and then purified by reverse-phase preparative HPLC using a 15% to 55% MeCN gradient in water with 0.1% formic acid modifier. The appropriate fractions were combined and lyophilized from a MeCN / water mixture to give Example 23 (14 mg, 34% yield) as a beige solid.

[0213] Other examples of inhibitors prepared in a similar manner are listed under Method B in Table 3.

[0214] General Method for the Synthesis of Inhibitors of Formula W-III (Method C) - Synthesis of Example 8: [ka]

[0215] Step 1: Preparation of substituted benzimidazole intermediate C-2: Bromobenzimidazole C-1 (70 mg, 0.355 mmol), potassium carbonate (196 mg, 1.42 mmol), and 3-pyridylboronic acid (57 mg, 0.46 mmol) were placed in a 4 mL vial, and dioxane (2 mL) and water (0.7 mL) were added. Argon gas was bubbled through the mixture for 1 min, and then tetrakis(triphenylphosphine)palladium(0) (16.4 mg, 0.014 mmol) was added. Argon gas was again bubbled through the solution for 3 min, and the vial was sealed and heated at 100 °C for 2 h (conversion to the desired product was complete as judged by LCMS analysis). The reaction mixture was cooled to room temperature, diluted with EtOAc, and washed with brine. After drying over MgSO, the extract was concentrated under reduced pressure and the residue was purified by flash chromatography using EtN pre-treated silica and a DCM-20% iPrOH / DCM gradient to provide the desired benzimidazole intermediate (58 mg, 84% yield): 1 H NMR(DMSO-d6) δ:12.71(bs,1H),9.24(s.1H),8.57(dd,J=5.1,1.6Hz,1H),8.43(broad d,J=5.5Hz,1H),8.31(s,1H),7.61(d,J=7.8Hz,1H),7.52(ddd,J=7.8,4.7,0.8Hz,1H),7.47(d,J=7.4Hz,1H),7.34(t,J=7.8Hz,1H). MS m / z 196.1(MH + ).

[0216] Step 2: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 was carried out as described above in Example 28 (Step 1).

[0217] Step 3: Preparation of W-III (Example 8): The substituted benzimidazole from Step 1 was coupled to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2-methyl-3-fluorophenyl) using DIPEA in NMP as described for Example 4 (Step 7) in General Method A.

[0218] Other compounds (i.e., Examples 9 and 10) were prepared in a similar manner using the appropriate intermediate A-11 (Ar = 2,3-dichlorophenyl or 2-methyl-3-chlorophenyl) and are listed under Method C in Table 3.

[0219] General Method for the Synthesis of Inhibitors of Formula W-IV (Method D) - Synthesis of Example 12: [ka]

[0220] Step 1: 3-Indole sulfonyl chloride D-2 was prepared as described in Org. Lett. 2011, 13, 3588. A solution of indole (3 g, 25.6 mmol) and sulfur trioxide-pyridine (4.08 g, 25.6 mmol) in pyridine (15 mL) was heated to reflux (115 °C) under stirring for 2 h. After 2 h, the reaction was cooled to room temperature and diluted with water (20 mL). The aqueous layer was washed twice with diethyl ether (20 mL). The aqueous layer was evaporated to dryness to give crude pyridinium 1H-indole-3-sulfonate D-1 (5.30 g, 75% yield) as a white solid. The crude material was used directly in the next step.

[0221] Step 2: Crude pyridinium 1H-indole-3-sulfonate (4.60 g, 16.7 mmol) from Step 1 was dissolved in a 1:1 mixture of sulfolane:acetonitrile (50 mL). The white suspension was cooled to 0 °C, and POCl (3.42 mL, 36.7 mmol) was added dropwise with stirring, resulting in a pale brown solution. The reaction was heated at 70 °C for 1 h. After 1 h, the orange solution was cooled to 0 °C. The cooled orange solution was added dropwise to 250 mL of ice water. A white precipitate formed during the addition. The solid was filtered, washed with water, and dried under vacuum to give 1H-indole-3-sulfonyl chloride D-2 (740 mg, 21% yield) as a gray solid.

[0222] Step 3: 1H-Indole-3-sulfonyl chloride (0.40 g, 1.86 mmol) was placed in a 25 mL flask and 6 mL of THF was added. The solution was cooled to 0 °C, and then 1-methylpiperazine (0.41 mL, 3.71 mmol) was added dropwise. The mixture changed from a red solution to a pale yellow solution with a large amount of gummy solid at the bottom of the flask. DIPEA (0.97 mL, 5.56 mmol) was then added, and the mixture was allowed to warm to room temperature. After stirring at room temperature for 30 minutes, the mixture was diluted with EtOAc and water. 3-4 mL of saturated ammonium chloride solution was added (pH of the aqueous layer approximately 7), and the layers were separated. The aqueous layer was extracted twice more with EtOAc, and the combined organic layers were washed once with water and then with brine. They were then dried over MgSO4, filtered, and concentrated to dryness. The resulting viscous oil was treated with a 2:1 mixture of hexane and diethyl ether (5-6 mL), sonicated (to give a slightly gummy solid), and the liquid was decanted. The residue was dried under reduced pressure to give 3-((4-methylpiperazin-1-yl)sulfonyl)-1H-indole D-3 (344 mg, 66% yield) as a beige foam, which was used directly without further purification. MS m / z 280.1 (MH + ). 1H NMR(DMSO-d6) δ:12.20(br.s.,1H),7.95(d,J=2.7Hz,1H),7.78(d,J=8.2Hz,1H),7.53(d,J=8.2 Hz,1H),7.26(td,J=7.6,1.2Hz,1H),7.20(ddd,J=8.2,7.0,1.2Hz,1H),2.89(br. s.,4H),2.34(t,J=4.9Hz,4H),2.10(s,3H).

[0223] Step 4: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 was carried out as described above in Method B for Example 28 (Step 1).

[0224] Step 5: Preparation of W-IV (Example 12): The sulfonamide intermediate D-3 (3-((4-methylpiperazin-1-yl)sulfonyl)-1H-indole) was coupled to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) using DIPEA in NMP as described for Example 4 (Step 7) in General Method A.

[0225] Other examples of inhibitors prepared in a similar manner are listed under Method D in Table 3.

[0226] General Method for the Synthesis of Inhibitors of Formula W-IV (Method E) - Synthesis of Example 70: [ka]

[0227] Step 1: Preparation of 3-indole sulfonamide fragment E-2: To a vial with a stir bar was added 1-(p-tolylsulfonyl)indole-3-sulfonyl chloride (117 mg, 0.316 mmol) (Chemical and Pharmaceutical Bulletin 2009, 57, 591) in anhydrous THF (1.5 mL). The solution was cooled to 0 °C, and DIEA (0.11 mL, 0.633 mmol) was added dropwise. Next, N-(2-methoxyethyl)ethylamine (0.039 mL, 0.316 mmol) was added dropwise, and the reaction mixture was allowed to warm gently to room temperature. The reaction was stirred at room temperature for 1 hour. Upon completion, 10% aqueous KOH was added dropwise (same volume as solvent). The reaction was heated at 60 °C overnight. Upon completion, the reaction mixture was diluted with EtOAc, and aqueous NH₄Cl was added. The layers were separated. The organic layer was washed with brine, then dried over MgSO, filtered, and concentrated to dryness to give the expected sulfonamide derivative (86 mg, 96%) as a pale orange oil. MS m / z 283.2 (MH + ).

[0228] Step 2: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 was carried out as described above in Method B for Example 28 (Step 1).

[0229] Step 3: Preparation of W-IV (Example 70): The sulfonamide intermediate E-2 (N-ethyl-N-(2-methoxyethyl)-1H-indole-3-sulfonamide) was coupled to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) using DIPEA in DMSO as described for Example 4 (Step 7) in General Method A.

[0230] Other examples of inhibitors prepared in a similar manner are listed under Method E in Table 3.

[0231] General Method for the Synthesis of Inhibitors of Formula WV (Method F) - Synthesis of Example 63: [ka]

[0232] Step 1: Preparation of indole-3-thioether (R = Et) fragment F-2: To a solution of 1H-indol-3-yl thiocyanate (Phosphorus, Sulfur and Silicon and the Related Elements 2014, 189, 1378) (175 mg, 1.0 mmol) in IPA (3.0 mL) was added sodium sulfide anhydrate (724 mg, 3.0 mmol) dissolved in water (0.5 mL), and the mixture was stirred at 50 °C for 2 h. Iodoethane (0.12 mL, 1.5 mmol) was then added to the resulting mixture, and the mixture was stirred at 50 °C overnight. The mixture was then diluted with DCM. The organic layer was washed with a saturated aqueous solution of ammonium chloride, followed by brine, dried over MgSO, filtered, and concentrated in vacuo to give crude sulfide F-2 (170 mg, 95%), which was used directly in the next step without further purification. MS m / z 178.2 (MH + ).

[0233] Step 2: Preparation of indole-3-ethylsulfone (R = Et) intermediate F-3: To a solution of 3-ethylsulfanyl-1H-indole (170 mg, 0.96 mmol) in DCM (4.8 mL) at room temperature was added 3-chloroperoxybenzoic acid (537 mg, 2.40 mmol). After stirring at this temperature for 4 h, the mixture was diluted with EtOAc, and the resulting solution was washed twice with a saturated aqueous solution of NaHCO3, followed by brine. The organic layer was then dried over MgSO4, filtered, and then concentrated in vacuo. The residual 3-ethylsulfonyl-1H-indole F-3 (90 mg, 45% yield) was then dissolved in DMSO and used directly in the next step (Step 4) without further purification. MS m / z 210.2 (MH + ).

[0234] Step 3: Preparation of the mixture of methyl sulfone and methyl sulfoxide of intermediate A-11 was carried out as described above in Method B for Example 28 (Step 1).

[0235] Step 4: Preparation of WV (Example 63): 3-Ethylsulfonyl-1H-indole intermediate F-3 was coupled to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) using DIPEA as described for Example 4 (Step 7) in General Method A.

[0236] Other examples of inhibitors prepared in a similar manner are listed under Method F in Table 3.

[0237] General Method for Inhibitors of Formula W-VI (Method G) - Synthesis of Example 113: [ka]

[0238] Step 1: 4-Methylpiperidin-4-ol (0.24 g, 1.84 mmol) and potassium carbonate (0.49 g, 3.52 mmol) were added to a solution of 3-fluoro-2-nitro-aniline (0.25 g, 1.60 mmol) in MeCN (2.6 mL). The resulting mixture was stirred at 85 °C for 10 h. MeCN was removed under reduced pressure, and EtOAc was added. The suspension was centrifuged, and the supernatant was poured into a flask. The solution was concentrated, and crude 1-(3-amino-2-nitro-phenyl)-4-methyl-piperidin-4-ol (0.40 g, 94% yield) was used for the next step without further purification. MS m / z 252.2 (MH + ).

[0239] Step 2: Iron (0.37 g, 6.70 mmol) and ammonium chloride (0.36 g, 6.70 mmol) were added to a mixture of 1-(3-amino-2-nitro-phenyl)-4-methyl-piperidin-4-ol (0.34 g, 1.34 mmol) in iPrOH (6.5 mL) and formic acid (1.9 mL, 49.6 mmol). The resulting mixture was heated to 90° C. and stirred for 10 minutes. The reaction mixture was cooled to room temperature and then filtered over Celite. (登録商標) The solution was concentrated and the crude product was purified by column chromatography (silica gel, 0-15% MeOH in DCM) to give 1-(1H-benzimidazol-4-yl)-4-methyl-piperidin-4-ol (0.17 g, 55% yield) as a reddish foamy solid. MS m / z 232.2 (MH + ). 1 H NMR(400MHz,DMSO-d6) δ:12.21(br.s,1H),8.02(s,1H),6.85-7.20(m,2H),6.33~6.67(m,1H),4.24(s,1H) ,3.15~3.26(m,2H),2.48(td,J=1.66,3.72Hz,2H),1.41~1.74(m,4H),1.16(s,3H).

[0240] Step 3: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 was carried out as described above in Method B for Example 28 (Step 1) (Ar = 2,3-dichlorophenyl).

[0241] Step 4: Preparation of W-VI (Example 113): 1-(1H-benzimidazol-4-yl)-4-methyl-piperidin-4-ol intermediate G-3 was coupled to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using cesium carbonate in DMSO in a similar manner as used for Example 4 (Step 7) in General Method A.

[0242] Other examples of inhibitors prepared in a similar manner are listed under Method G in Table 3.

[0243] Preparation of Examples 17, 47 and 99: [ka]

[0244] Step 1: Preparation of 4-(trifluoromethyl)-1H-benzo[d]imidazole (for Example 17): In a 10 mL vial, 3-(trifluoromethyl)benzene-1,2-diamine (280 mg, 1.59 mmol) in formic acid (2.0 mL) was added. The reaction was heated to 100° C. and stirred for 3 hours. The reaction was cooled to room temperature, and the formic acid was removed under reduced pressure. The residue was taken up in a saturated solution of NaHCO to give a precipitate, which was homogenized by sonication. The solid was collected by filtration, washed with water, briefly dried on the filter, and collected. The product was dried under reduced pressure overnight to give 4-(trifluoromethyl)-1H-benzimidazole (269 mg, 91% yield) as a light brown solid. MS m / z 187.2 (MH + ). 1 H NMR (DMSO-d6) δ:12.93(br.s.,1H),8.39(s,1H),7.88(d,J=8.2Hz,1H),7.54(d,J=7.8Hz,1H),7.36(t,J=7.8Hz,1H).

[0245] 5,6-Dichloro-1H-benzimidazole (for Example 47) was prepared in a similar manner starting from commercially available 4,5-dichloro-1,2-phenylenediamine. The product was obtained as a pale brown solid (100% yield). MS m / z 187.0 (MH + ). 1 H NMR (chloroform-d) δ: 9.66 (br.s., 1H), 8.09 (s, 1H), 7.78 (br.s., 2H).

[0246] 4-Methoxy-1H-benzimidazole (for Example 99) was prepared in a similar manner starting from commercially available 3-methoxybenzene-1,2-diamine. The product was obtained as an off-white solid after silica gel chromatography (EtOAc / heptane) (71% yield). MS m / z 149.2 (MH + ).

[0247] Step 2: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 was carried out as described above in Method B for Example 28 (Step 1) (Ar = 2,3-dichlorophenyl).

[0248] Step 3. Examples 17, 47, and 99 were prepared by coupling the appropriate substituted benzimidazole to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using DIPEA in NMP as described for Example 4 (Step 7) in General Method A.

[0249] Preparation of Examples 49 and 71: [ka]

[0250] Step 1: Preparation of 4,5-difluoro-1H-benzimidazole (for Example 49): 2,3-Difluoro-6-nitro-aniline (800 mg, 4.60 mmol) was placed in a flask along with ammonium chloride (2.46 g, 46.0 mmol) and iron powder (2.57 g, 46.0 mmol). Next, IPA (11.5 mL) was added, followed by formic acid (12 mL, 305 mmol). The flask was fitted with a reflux condenser, and the reaction flask was immersed in an oil bath set at 80° C. and stirred for 5 hours. After cooling, the mixture was transferred to a Celite tube rinsed with IPA. (登録商標)The mixture was filtered through a small pad of hexane, and the filtrate was concentrated to dryness. The residue was then neutralized with a saturated solution of NaHCO3 and extracted three times with EtOAc. The combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated. The resulting residue was purified by silica gel chromatography using a 50% to 100% EtOAc gradient in DCM. The appropriate fractions were pooled, concentrated, and dried under reduced pressure to give 4,5-difluoro-1H-benzimidazole (706 mg, 99% yield). MS m / z 155.2 (MH + ). 1 H NMR (DMSO-d6) δ:13.26(br.s.,0.2H),12.83(br.s.,0.8H),8.32(s,1H),7.43~7.54(m,0.2H),7.34(dd,J=9.0,3.9Hz,0.8H),7.17~7.31(m,1H).

[0251] 4,6-Difluoro-1H-benzimidazole (as in Example 71) was prepared in a similar manner (93% yield) starting from commercially available 2,4-difluoro-6-nitro-aniline. MS m / z 155.2 (MH + ). 1 H NMR (DMSO-d6) δ:12.87(br.s.,1H),8.28(s,1H),7.25(d,J=7.8Hz,1H),7.05(t,J=10.4Hz,1H).

[0252] Step 2: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 was carried out as described above in Method B for Example 28 (Step 1) (Ar = 2,3-dichlorophenyl).

[0253] Step 3: Examples 49 and 71 were prepared by coupling the appropriate substituted benzimidazole described above to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using DIPEA in NMP as described for Example 4 (Step 7) in General Method A.

[0254] Preparation of Example 72: [ka]

[0255] Step 1: Preparation of 3-methylsulfonyl-1H-indole: Indole (182 mg, 1.55 mmol) was dissolved in THF (17 mL) under nitrogen at room temperature. Potassium tert-butoxide (192 mg, 1.71 mmol) was added, and the mixture was stirred for 30 minutes. Triethylborane (0.22 mL, 1.55 mmol) was added dropwise at room temperature and stirred for 30 minutes. Methanesulfonyl chloride (0.13 mL, 1.71 mmol) was added, and the mixture was stirred at −15° C. for 5 days. Next, a saturated aqueous solution of NaHCO was added, and the mixture was extracted with EtOAc. The organic layer was washed with brine, dried over MgSO, filtered, and concentrated in vacuo. The residue was purified by chromatography on silica gel using an EtOAc gradient in hexanes (20-60%) to give 3-methylsulfonyl-1H-indole (30 mg, 10% yield).

[0256] Step 2: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 was carried out as described above in Method B for Example 28 (Step 1) (Ar = 2,3-dichlorophenyl).

[0257] Step 3: Example 72 was prepared by coupling the above-described 3-methylsulfonyl-1H-indole with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using DIPEA in DMSO as described for Example 4 (Step 7) in General Method A.

[0258] Preparation of Examples 79 and 80: [ka]

[0259] Step 1: Preparation of 2-fluoro-3-(2-methoxyethoxy)-6-nitroaniline (Example 79): 2,3-Difluoro-6-nitroaniline (800 mg, 4.60 mmol) was placed in a 25 mL flask and dissolved in 10 mL of DMF at room temperature to give a bright yellow solution. Next, 2-methoxyethanol (2.2 mL, 27.6 mmol) was added, followed by potassium carbonate (2.54 g, 18.4 mmol). The color of the solution changed to bright red. The mixture was heated in an oil bath at 80° C. for 22 hours. After cooling, the mixture was diluted with EtOAc and washed with saturated aqueous ammonium chloride solution. The aqueous layer was extracted twice with EtOAc, and the combined organic layers were washed with brine, dried over MgSO4, filtered, and concentrated. The resulting residue was purified by silica gel chromatography using an EtOAc gradient in hexanes to give 2-fluoro-3-(2-methoxyethoxy)-6-nitro-aniline (1.01 g, 96% yield). 1 H NMR (DMSO-d6) δ:7.87(dd,J=9.8,2.0Hz,1H),7.16(br.s,2H),6.60(dd,J=10.0,8.0Hz,1H),4.24~4.32(m,2H),3.63~3.71(m,2H),3.31(s,3H).

[0260] 2-(3-amino-2-fluoro-4-nitro-phenoxy)ethanol (for Example 80) was prepared in a similar manner using ethylene glycol. The product was obtained in 98% yield. 1 H NMR(DMSO-d6) δ:7.86(dd,J=9.8,2.0Hz,1H),7.15(s,2H),6.60(dd,J=9.8,7.8Hz,1H),4.94(t,J=5.5Hz,1H),4.17(t,J=4.9Hz,2H),3.73(q,J=4.8Hz,2H).

[0261] Step 2: Preparation of substituted benzimidazoles from the 2-nitroanilines described in Step 1 was accomplished using the "one-pot" procedure described in Method G for Example 113 (Step 2).

[0262] 4-Fluoro-5-(2-methoxyethoxy)-1H-benzimidazole (for Example 79): 82% yield. MS m / z 211.2 (MH + ). 1 H NMR(DMSO-d6) δ:12.90(br.s.,0.3H),12.57(br.s.,0.7H),8.20(s,0.7H),8.18(s,0.3H),7.39(d,J=8.6Hz,0.3H),7.25(d,J=8.6Hz,0.7H),7. 09(t,J=8.2Hz,0.7H),7.06(t,J=8.2Hz,0.3H),4.19(t,J=4.3Hz,0.6H),4.16(t,J=4.3Hz,1.4H),3.62~3.70(m,2H),3.32(s,3H).

[0263] 2-[(4-Fluoro-1H-benzimidazol-5-yl)oxy]ethanol (for Example 80): 53% yield. MS m / z 197.2 (MH + ). 1 H NMR(DMSO-d6) δ:12.88(br.s.,0.3H),12.56(br.s.,0.7H),8.19(s,0.7H),8.17(s,0.3H),7.39(d,J=9.0Hz,0.3H),7.24(d,J=8.6Hz,0.7H),7.10(t,J =8.0Hz,0.7H),7.06(t,J=8.2Hz,0.3H),4.87(t,J=5.5Hz,1H),4.09(t,J=4.7Hz,0.6H),4.06(t,J=5.1Hz,1.4H),3.71(q,J=5.2Hz,2H).

[0264] Step 3: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 was carried out as described above in Method B for Example 28 (Step 1) (Ar = 2,3-dichlorophenyl).

[0265] Step 4: Examples 79 and 80 were prepared by coupling the appropriate substituted benzimidazole described above to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using DIPEA in NMP as described for Example 4 (Step 7) in General Method A.

[0266] Preparation of Example 81: [ka]

[0267] Step 1: Preparation of (5-chloro-1H-indol-3-yl)-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)methanone: 5-Chloro-1H-indole-3-carboxylic acid (50 mg, 0.256 mmol) was dissolved in NMP (1.5 mL). 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (98 mg, 0.31 mmol) was added, followed by 3-oxa-8-aza-bicyclo[3.2.1]octane HCl (42 mg, 0.28 mmol) at room temperature. N,N-Diisopropylethylamine (0.18 mL, 1.02 mmol) was finally added, and the mixture was stirred for 1 hour. It was then partitioned between EtOAc and a saturated aqueous solution of ammonium chloride. The layers were separated, and the aqueous layer was further extracted twice with EtOAc. The combined organic layers were washed with brine, then dried over MgSO4, filtered, and concentrated. The resulting residue was purified by silica gel chromatography using a 1:1 EtOAc in DCM to 100% EtOAc gradient, followed by 2% IPA in EtOAc to give (5-chloro-1H-indol-3-yl)-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)methanone (65 mg, 88% yield). MS m / z 291.1 (MH + ). 1H NMR(DMSO-d6) δ:11.83(br.s.,1H),7.89(s,1H),7.83(d,J=2.0Hz,1H),7.46(d,J=8.6Hz,1H),7.17(dd,J=8.6,2. 3Hz, 1H), 4.47 (br.s., 2H), 3.68 (d, J = 11.0Hz, 2H), 3.61 (dd, J = 10.6, 1.2Hz, 2H), 1.80~1.94 (m, 4H).

[0268] Step 2: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 was carried out as described above in Method B for Example 28 (Step 1) (Ar = 2,3-dichlorophenyl).

[0269] Step 3: Example 81 was prepared by coupling the above-described (5-chloro-1H-indol-3-yl)-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)methanone to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using DIPEA in DMSO as described for Example 4 (Step 7) in General Method A.

[0270] Preparation of Examples 91, 92, 106 and 107: [ka]

[0271] Step 1: Preparation of 1-(1H-indol-3-yl)propan-1-one (for Example 91): To a 100 mL round-bottom flask was added indole (200 mg, 1.71 mmol) in 7 mL of DCM. The solution was cooled to 0° C., and then a 1.8 M (25 wt.) solution of EtAlCl in toluene (1.42 mL, 2.56 mmol) was added dropwise. The reaction was stirred for 30 minutes at 0° C. Propionyl chloride in 6 mL of DCM was added dropwise. The reaction was warmed to room temperature and stirred for 2.5 hours. The reaction was quenched with 3 equivalents of NaOAc in 5 mL of water. The reaction was diluted with DCM and evaporated on Celite.(登録商標) The aluminum salts were removed by filtration through a separatory funnel. The filtrate was transferred to a separatory funnel, and then water was added. 1 M NaOH was added to dissolve the remaining aluminum salts. The layers were separated, and the organic layer was washed with water and then brine. The organic layer was then dried over MgSO4, filtered, and then concentrated under reduced pressure. The crude material was purified by silica gel chromatography using a gradient from 100% hexane to 75% EtOAc in hexane to give 1-(1H-indol-3-yl)propan-1-one (228.9 mg, 77% yield) as a pale yellow solid. MS m / z 174.2 (MH + ). 1 H NMR(400MHz,DMSO-d6) δ:11.88(br.s.,1H),8.30(s,1H),8.15~8.23(m,1H),7.41~7.49(m,1H),7.18(qui nt,J=7.2,7.2,7.2,7.2,1.4Hz,2H),2.87(q,J=7.4Hz,2H),1.11(t,J=7.4Hz,3H).

[0272] 1-(1H-indol-3-yl)-2-methyl-propan-1-one (for Example 92) was prepared in a similar manner using isobutyric acid chloride. The product was obtained in 94% yield. MS m / z 188.2 (MH + ). 1 H NMR(400MHz,DMSO-d6) δ:11.90(br.s.,1H),8.35(s,1H),8.20(dd,J=8.0,1.4Hz,1H),7.46(dd,J=7.4,1.2Hz,1H),7 .18(quint,J=7.3,7.3,7.3,7.3,1.2Hz,2H),3.45(spt,J=6.7Hz,1H),1.12(d,J=6.7Hz,6H).

[0273] 1-(1H-indol-3-yl)-3-methoxy-propan-1-one (for Example 106) was prepared in a similar manner using 3-methoxypropanoyl chloride. The product was obtained in 86% yield. MS m / z 204.2 (MH+ ).

[0274] 1H-Indol-3-yl(2-thienyl)methanone (as in Example 107) was prepared in a similar manner using 2-thiophenecarbonyl chloride. The product was obtained in 80% yield. MS m / z 228.0 (MH + ).

[0275] Step 2: Preparation of the mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 was carried out as described above in Method B for Example 28 (Step 1) (Ar = 2,3-dichlorophenyl).

[0276] Step 3: Examples 91, 92, 106, and 107 were prepared by coupling the appropriate substituted indole described above to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using DIPEA in DMSO as described for Example 4 (Step 7) in General Method A.

[0277] Preparation of Example 98: [ka]

[0278] Step 1: tert-Butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate (0.70 g, 3.30 mmol) and potassium carbonate (1.14 g, 8.24 mmol) were placed in a 25 mL flask and suspended in 10 mL of MeCN. Benzyl 2-bromoethyl ether (0.57 mL, 3.63 mmol) was then added dropwise at room temperature. The resulting mixture was stirred at room temperature for 22 hours and then at 40 °C for 4 hours. The reaction mixture was diluted with EtOAc. The solids were removed by filtration, and the filtrate was concentrated to dryness. The resulting residue was purified by flash chromatography on silica gel using a 0% to 40% EtOAc gradient in hexanes. The appropriate fractions were then concentrated and then dried under reduced pressure to give tert-butyl 3-(2-benzyloxyethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (678 mg, 59% yield) as a colorless oil. MS m / z 347.4 (MH + ). 1 H NMR(DMSO-d6) δ:7.24~7.39(m,5H),4.47(s,2H),3.99(br.s.,2H),3.51(t,J=5.7Hz,2H),2.64(d,J=10 .2Hz,2H),2.50(t,J=5.7Hz,2H),2.19(d,J=10.6Hz,2H),1.60~1.82(m,4H),1.39(s,9H).

[0279] Step 2: tert-Butyl 3-(2-benzyloxyethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (673 mg, 1.94 mmol) was placed in a 100 mL flask and dissolved in 5 mL of dioxane. Next, a 4 N solution of HCl in dioxane (1.94 mL, 7.76 mmol) was added dropwise. After stirring for 1 h, another portion of the HCl solution (2.91 mL, 11.64 mmol) was added, and the mixture was warmed to 40 °C. After stirring at that temperature for an additional 2 h, a gum separated at the bottom of the flask. The mixture was cooled to room temperature, diluted with methanol, and concentrated to dryness. The residue was dissolved in methanol and concentrated to dryness, then dried under reduced pressure again. This gave 3-(2-benzyloxyethyl)-3,8-diazabicyclo[3.2.1]octane dihydrochloride (620 mg, 100% yield) as a white gummy solid. MS m / z 247.3 (MH + ). 1 H NMR (DMSO-d6) δ: 11.36 (br.s., 1H), 10.10 (br.s., 1H), 9.64 (br.s., 1H), 7.33–7.40 (m, 4H), 7.27–7.33 (m, 1H), 4.51 (s, 2H), 4.18 (br.s., 2H), 3.88 (br.s., 2H), 3.58–3.75 (m, 2H), 3.21 (br.s., 2H), 2.34 (br.s., 2H), 2.02 (br.s., 2H) (2H hidden underwater peak).

[0280] Step 3: In a 100 mL flask, to a solution of 3-(2-benzyloxyethyl)-3,8-diazabicyclo[3.2.1]octane; dihydrochloride (615 mg, 1.93 mmol) in 15 mL of methanol was added 10% palladium on charcoal (205 mg, 0.193 mmol). The flask was placed under reduced pressure and then filled with hydrogen. This procedure was repeated three more times, and then the mixture was stirred vigorously under a balloon atmosphere of hydrogen over the weekend. The flask was flushed with nitrogen, and the reaction mixture was transferred to Celite. (登録商標)The mixture was filtered through a short pad of hexane. The filtrate was concentrated and dried under reduced pressure to give 2-(3,8-diazabicyclo[3.2.1]octan-3-yl)ethanol dihydrochloride (406 mg, 92% yield). MS m / z 157.2 (MH + ). 1 H NMR (DMSO-d6) δ: 11.14 (br.s., 1H), 10.07 (br.s., 1H), 9.68 (br.s., 1H), 5.08 (br.s., 1H), 4.16 (br.s., 2H), 3.81 (br.s., 2H), 3.03 (br.s., 2H), 2.33 (br.s., 2H), 2.03 (br.s., 2H) (4H hidden underwater peak).

[0281] Step 4: Example 98 was prepared by coupling the above-described amine to intermediate B-1 (Ar=2,3-dichlorophenyl) in a similar manner as described in General Method B for Example 28 (Step 4), using TBTU instead of HATU.

[0282] Preparation of Example 100: [ka]

[0283] Step 1: Preparation of 6-bromo-4-fluoro-1H-benzimidazole: This compound was prepared in a similar manner to the benzimidazole described for Example 49 / 71 (Step 1) using iron, ammonium chloride, and formic acid, starting with commercially available 4-bromo-2-fluoro-6-nitro-aniline. After silica gel chromatography (EtOAc / DCM gradient), 6-bromo-4-fluoro-1H-benzimidazole was obtained as an off-white solid (96% yield). MS m / z 215.0 (MH + ). 1H NMR (DMSO-d6) δ:12.94(br.s.,1H),8.31(s,1H),7.63(s,1H),7.28(d,J=10.6Hz,1H).

[0284] Step 2: Preparation of 2-[(6-bromo-4-fluoro-benzimidazol-1-yl)methoxy]ethyl-trimethyl-silane and 2-[(5-bromo-7-fluoro-benzimidazol-1-yl)methoxy]ethyl-trimethyl-silane: 6-Bromo-4-fluoro-1H-benzimidazole (500 mg, 2.05 mmol) was dissolved in 10 mL of DMF, and then potassium carbonate (848 mg, 6.14 mmol) was added, followed by SEM-Cl (0.44 mL, 2.46 mmol) at room temperature. The mixture was stirred at the same temperature for 18 hours. An additional portion of SEM-Cl (0.18 mL, 1.03 mmol) was added, and the mixture was stirred for another hour. The mixture was poured into a saturated solution of ammonium chloride and then extracted three times with EtOAc. The combined organic layers were washed with water, then brine, dried over MgSO4, filtered, and concentrated. The resulting residue was purified by silica gel chromatography using a gradient of 100% hexane to 40% EtOAc in hexane. The appropriate fractions were pooled, concentrated, and dried under reduced pressure to give a 3:1 mixture of 2-[(6-bromo-4-fluoro-benzimidazol-1-yl)methoxy]ethyl-trimethyl-silane and 2-[(5-bromo-7-fluoro-benzimidazol-1-yl)methoxy]ethyl-trimethyl-silane (689 mg, 97% yield) as a waxy beige solid. MS m / z 385.2 (MH + ). 1H NMR(DMSO-d6) δ:8.50(s,0.25H),8.46(s,0.75H),7.80(d,J=1.6Hz,0.75H),7.77(d,J=1.6Hz,0.25H),7.43(dd,J=10.6,1.6Hz,0.25H),7.36(dd,J=10.6,1.6Hz,0.75H),5.66(s,1.5H),5.64(s,0.5H),3.45~3.54(m,2H),0.78~0.86(m,2H),-0.09(s,6.75H),-0.11(s,2.25H)。

[0285] Step 3: Preparation of 2-[7-fluoro-3-(2-trimethylsilylethoxymethyl)benzimidazol-5-yl]oxyethanol: The 3:1 isomeric mixture of 2-[(6-bromo-4-fluorobenzimidazol-1-yl)methoxy]ethyl-trimethylsilane and 2-[(5-bromo-7-fluorobenzimidazol-1-yl)methoxy]ethyl-trimethylsilane (630 mg, 1.82 mmol) described above was placed in a 20 mL vial, followed by 1,10-phenanthroline (33 mg, 0.18 mmol), copper(I) iodide (35 mg, 0.18 mmol), and cesium carbonate (1.19 g, 3.65 mmol). Ethylene glycol (8.2 mL, 147 mmol) was added. The resulting mixture was stirred at 120 °C for 21 hours. The mixture was cooled to room temperature, and then it was diluted with a saturated solution of ammonium chloride, water, and EtOAc. The layers were separated, and the aqueous layer was further extracted twice with EtOAc. The combined organic layers were diluted with a small amount of hexane and washed once with a saturated aqueous solution of ammonium chloride, twice with water, and once with brine. The organic layer was then dried over MgSO, filtered, and concentrated. The residue was purified by silica gel chromatography using a 30% to 90% EtOAc in hexane gradient. The appropriate fractions were pooled, concentrated, and dried to afford 2-[7-fluoro-3-(2-trimethylsilylethoxymethyl)benzimidazol-5-yl]oxyethanol (307 mg, 52% yield) as a thick, colorless oil, which slowly crystallized after drying. MS m / z 327.3 (MH + ). 1 H NMR(DMSO-d6) δ:8.26(s,1H),7.06(d,J=2.0Hz,1H),6.73(dd,J=12.3,2.2Hz,1H),5.60(s,2H),4.89(t,J=5.5Hz,1H), 4.03(t,J=5.1Hz,2H),3.74(q,J=5.5Hz,2H),3.49(t,J=7.8Hz,2H),0.83(t,J=7.8Hz,2H),-0.09(s,9H).

[0286] Step 4: Preparation of 2-[(7-fluoro-3H-benzimidazol-5-yl)oxy]ethanol: 2-[7-Fluoro-3-(2-trimethylsilylethoxymethyl)benzimidazol-5-yl]oxyethanol (308 mg, 0.94 mmol) was dissolved in 5 mL of THF in a 20 mL vial, and then a 1 M THF solution of tetra-N-butylammonium fluoride (4.72 mL, 4.72 mmol) was added at room temperature. The vial was then sealed, and the mixture was heated at 65° C. for 23 hours. The mixture was diluted with EtOAc and water with a small amount of NaHCO. The layers were separated, and the aqueous layer was further extracted four times with EtOAc. The combined organic layers were washed with brine, then dried over MgSO, filtered, and concentrated. The gummy solid residue was purified by silica gel chromatography using a 2% to 20% IPA gradient in EtOAc. The product-containing fractions were pooled, concentrated, and dried under reduced pressure to give 2-[(7-fluoro-3H-benzimidazol-5-yl)oxy]ethanol (139 mg, 75% yield) as a white solid. 1 H NMR showed an 85:15 mixture of tautomers. MS m / z 197.2 (MH + ). 1 H NMR(DMSO-d6) δ:12.89(br.s.,0.15H),12.51(br.s.,0.85H),8.18(br.s.,0.15H),8.10(s,0.85H),7.03(br.s.,0.15H),6.86(br.s.,0. 85H),6.73(br.s.,0.15H),6.66(d,J=12.1Hz,0.85H),4.88(t,J=5.5Hz,1H),4.01(t,J=4.9Hz,2H),3.72(q,J=5.3Hz,2H).

[0287] Step 5: Preparation of the mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 was carried out as described above in Method B for Example 28 (Step 1) (Ar = 2,3-dichlorophenyl).

[0288] Step 6: Example 100 was prepared by coupling the above-described 2-[(7-fluoro-3H-benzimidazol-5-yl)oxy]ethanol to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using DIPEA in DMSO as described for Example 4 (Step 7) in General Method A.

[0289] Preparation of Examples 104, 112, 116-118: [ka]

[0290] Step 1: To a 20 mL vial was added 1H-benzimidazole-4-carboxylic acid (1.00 g, 6.17 mmol) in MeOH (6.2 mL). Then, sulfuric acid (723 μL, 13.6 mmol) was added. The reaction was heated in an oil bath at 70° C. for 24 hours. It was cooled to room temperature. The solvent was removed under reduced pressure. The crude material was taken up in a saturated aqueous solution of NaHCO and extracted three times with EtOAc. The combined organic layers were washed with water, then brine, dried over MgSO, filtered, and then concentrated under reduced pressure to give methyl 1H-benzimidazole-4-carboxylate (895 mg, 5.08 mmol, 82% yield) as a brown solid, which was used without further purification. MS m / z 177.2 (MH + ).

[0291] Step 2: To a 250 mL round-bottom flask, methyl 1H-benzimidazole-4-carboxylate (750 mg, 4.26 mmol) from the previous step and 2-(trimethylsilyl)ethoxymethyl chloride, stabilized tech. (982 μL, 5.53 mmol) in anhydrous THF (21 mL) were added under a N atmosphere. The mixture was cooled to 0 °C, and then a 1 M solution of lithium bis(trimethylsilyl)amide in THF (5.5 mL, 5.5 mmol) was added slowly over 20 minutes. The reaction was allowed to warm to room temperature and stirred overnight. The reaction was cooled to 0 °C, and then a 1 M solution of lithium aluminum hydride in THF (5.1 mL, 5.1 mmol) was added. The reaction was allowed to warm to room temperature and stirred overnight. The reaction was quenched with MeOH at 0 °C. Next, it is Celite (登録商標) The resulting mixture was adsorbed onto a silica gel column and purified by normal-phase silica gel chromatography using a 0% to 10% IPA gradient in EtOAc. The desired fractions were collected and concentrated under reduced pressure to give [1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]methanol (461 mg, 39% yield) as a light brown solid. MS m / z 279.2 (MH + ). 1 H NMR(400MHz,DMSO-d6) δ:8.31(s,1H),7.50(d,J=9.0Hz,1H),7.24~7.32(m,2H),5.63(s,2H),5.14(t,J=5.5Hz ,1H),4.92(d,J=5.5Hz,2H),3.48(t,J=8.0Hz,2H),0.83(t,J=8.0Hz,2H),-0.09(s,9H).

[0292] Step 3: In a 20 mL vial, [1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]methanol (200 mg, 0.72 mmol) from the previous step and DIPEA (500 μL, 2.87 mmol) in MeCN (8 mL) were added. Methanesulfonyl chloride (167 μL, 2.16 mmol) was then added. The reaction was stirred overnight at room temperature. The reaction was quenched with the addition of a saturated aqueous solution of NaHCO and then extracted three times with EtOAc. The combined organic layers were dried over MgSO, filtered, and then concentrated under reduced pressure.

[0293] The crude material was dissolved in DMF (8 mL), and then potassium cyanide (187 mg, 2.87 mmol) was added. The reaction was again stirred at room temperature overnight. The reaction was quenched with the addition of a 10% aqueous solution of LiCl and then extracted three times with EtOAc. The combined organic layers were dried over MgSO4, filtered, and then concentrated under reduced pressure. The crude material was purified by evaporation over Celite 400°C. (登録商標) The crude product was adsorbed onto hexanes and then purified by normal-phase silica gel chromatography using a 0% to 100% EtOAc gradient in hexanes. The desired fractions were collected and concentrated under reduced pressure to give 2-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]acetonitrile (149 mg, 72% yield) as a clear film. MS m / z 288.2 (MH+).

[0294] Step 4: To sodium hydride (60 mg, 1.50 mmol) in a 20 mL vial was added 4 mL of DMF. The mixture was cooled to 0° C., and then a solution of 2-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]acetonitrile (177 mg, 0.616 mmol) from the previous step in 4 mL of DMF was added slowly. Upon addition, the reaction turned pale orange. The reaction was allowed to warm to room temperature. After approximately 30 minutes, the reaction turned a deeper orange. The reaction was cooled to 0° C., and then iodomethane (115 μL, 1.85 mmol) was added. Upon addition, the reaction turned pale orange. The reaction was allowed to warm to room temperature and stirred overnight. The reaction was quenched with 1M aqueous HCl and then extracted three times with EtOAc. The combined organic layers were washed with water, then brine, dried over MgSO, filtered, and then concentrated under reduced pressure. The crude material was purified by evaporation over Celite. (登録商標) The resulting mixture was adsorbed onto a silica gel column and purified by normal-phase silica gel chromatography using a 0% to 100% EtOAc gradient in hexanes. The desired fractions were collected and concentrated under reduced pressure to give 2-methyl-2-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]propanenitrile (134 mg, 69% yield) as an orange-brown oil. MS m / z 316.2 (MH + ).

[0295] Step 5: To a 20 mL scintillation vial was added 2-methyl-2-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]propanenitrile (134 mg, 0.426 mmol) from the previous step, DCM (2 mL), and TFA (2 mL). The reaction was stirred overnight at room temperature. The solvent was removed under reduced pressure. The material was dissolved in MeOH, and then Amberlite IRA-67 resin was added. The resin was filtered through a cotton plug. The organics were collected and concentrated under reduced pressure to give 2-(1H-benzimidazol-4-yl)-2-methyl-propanenitrile; 2,2,2-trifluoroacetic acid (123 mg, 96% yield) as a clear film. MS m / z 186.0 (MH + ).

[0296] Step 6: Preparation of a mixture of methyl sulfone and methyl sulfoxide from the appropriate intermediate A-11 was carried out as described above in Method B for Example 28 (Step 1).

[0297] Step 7: Examples 104, 112, 116-118 were prepared by coupling the benzimidazole described in Step 5 to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (with the appropriate Ar group) using DIPEA in DMSO as described for Example 4 (Step 7) in General Method A.

[0298] Preparation of Example 105: [ka]

[0299] Step 1: Preparation of 4-methoxy-1H-imidazo[4,5-c]pyridine: In a pressure-resistant tube, sodium hydride (104 mg, 2.60 mmol) was slowly added to 3.3 mL of MeOH at 0° C. The resulting mixture was stirred for 5 minutes, and commercially available 4-chloro-1H-imidazo[4,5-c]pyridine (0.10 g, 0.65 mmol) was added. The tube was sealed, and the reaction mixture was heated to 120° C. and stirred for 12 hours. After cooling to room temperature, the resulting mixture was concentrated under reduced pressure. The crude product was purified by silica gel chromatography using a gradient of 0% to 10% MeOH in DCM to give 4-methoxy-1H-imidazo[4,5-c]pyridine (51 mg, 53% yield) as a tan solid. MS m / z 150.2 (MH + ).

[0300] Step 2: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 (Ar = 2,3-dichlorophenyl) was carried out as described above in Method B for Example 28 (Step 1).

[0301] Step 3: Example 105 was prepared in a manner similar to that used for Example 4 (Step 7) in General Method A by coupling 4-methoxy-1H-imidazo[4,5-c]pyridine described in Step 1 to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using cesium carbonate in DMSO.

[0302] Preparation of Example 108: [ka]

[0303] Step 1: Preparation of 3-ethoxy-2-nitro-aniline: A solution of sodium ethoxide (21% in EtOH, 1.63 mL, 4.36 mmol) was added to a solution of 3-fluoro-2-nitro-aniline (0.23 g, 1.45 mmol) in EtOH (8 mL). The resulting mixture was stirred at 80 °C for 10 h. The reaction mixture was concentrated, and water was added. The aqueous mixture was extracted with EtOAc. The organic layers were combined, washed with brine, dried over NaSO, filtered, and concentrated. The crude product was purified by column chromatography (silica gel, 0-100% EtOAc in hexane) to give 3-ethoxy-2-nitro-aniline (0.25 g, 94% yield). MS m / z 183.0 (MH + ). 1 H NMR(400MHz,DMSO-d6) δ:7.10(t,J=8.4Hz,1H),6.42(dd,J=8.4,1.0Hz,1H),6.28(dd,J=8.2,1.2Hz,1H),5.94(br.s,2H),4.03(q,J=6.9Hz,2H),1.25(t,J=7.0Hz,3H).

[0304] Step 2: Preparation of 4-ethoxy-1H-benzimidazole: Iron (0.37 g, 6.70 mmol) and ammonium chloride (0.36 g, 6.70 mmol) were added to a mixture of 3-ethoxy-2-nitro-aniline (0.24 g, 1.34 mmol) from the previous step and formic acid (1.9 mL, 49.6 mmol) in iPrOH (4.0 mL). The resulting mixture was heated to 90 °C and stirred for 10 h. The reaction mixture was cooled to room temperature and filtered through a Celite (登録商標) The solution was concentrated and the crude product was purified by column chromatography (silica gel, 0-10% MeOH in DCM) to give 4-ethoxy-1H-benzimidazole (133 mg, 61% yield) as an off-white solid. MS m / z 163.0 (MH + ). 1H NMR(400MHz,DMSO-d6) δ:7.98~8.10(m,1H),7.97~8.21(m,2H),7.01~7.11(m,1H),6.70(d,J=7.4Hz,1H),4.23(q,J=7.0Hz,2H),1.27~1.50(m,3H).

[0305] Step 3: Preparation of a mixture of (Ar=2,3-dichlorophenyl)methyl sulfone and methyl sulfoxide from intermediate A-11 was carried out as described above in Method B for Example 28 (Step 1).

[0306] Step 4: Example 108 was prepared by coupling the benzimidazole described in Step 2 to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using cesium carbonate in DMSO in a manner similar to that used for Example 4 (Step 7) in General Method A.

[0307] Preparation of Example 110: [ka]

[0308] Step 1: Preparation of tert-butyl 3-iodoindole-1-carboxylate: In a 200 mL flask, indole (1.00 g, 8.54 mmol) was dissolved in 26 mL of DMF. Next, iodine (2.38 g, 9.39 mmol) and potassium hydroxide (1.20 g, 21.3 mmol) were added. The reaction was stirred at room temperature for 5 hours. Next, it was added to 100 mL of saturated aqueous NaSO solution. The mixture was then extracted three times with EtOAc. The combined organic layers were washed with water, then brine, dried over MgSO, filtered, and then concentrated under reduced pressure. To this crude material, 43 mL of THF and 4-dimethylaminopyridine (10 mg, 0.09 mmol) were added, followed by di-tert-butyl dicarbonate (2.05 g, 9.39 mmol). The resulting mixture was stirred at room temperature for 18 hours and then purified by silica gel chromatography using a 0% to 75% EtOAc gradient in hexanes. The appropriate fractions were pooled and concentrated to give tert-butyl 3-iodoindole-1-carboxylate (2.79 g, 95% yield) as a brown oil. 1 H NMR (400MHz, DMSO-d6) δ:8.07(d,J=8.2Hz,1H),7.89(s,1H),7.39~7.45(m,1H),7.32~7.39(m,2H),1.62(s,9H).

[0309] Step 2: Preparation of 1-(1H-indol-3-yl)pyrrolidin-2-one and tert-butyl 3-(2-oxopyrrolidin-1-yl)indole-1-carboxylate: A flask was charged with tert-butyl 3-iodoindole-1-carboxylate (100 mg, 0.29 mmol), 2-pyrrolidinone (74 mg, 0.87 mmol), cesium carbonate (0.285 g, 0.87 mmol), CuI (28 mg, 0.15 mmol), and N,N'-dimethylethylenediamine (31 μL, 0.29 mmol) in dioxane (1.5 mL). The resulting mixture was heated at 80 °C and stirred for 22 h. The reaction mixture was filtered through a Celite (登録商標)The mixture was filtered through a filtration media and the crude product was purified by column chromatography (silica gel, 0-100% EtOAc gradient in hexanes, followed by 0-10% methanol gradient in EtOAc) to give 1-(1H-indol-3-yl)pyrrolidin-2-one (11.9 mg, 20% yield) as a white solid (MS m / z 201.2) and tert-butyl 3-(2-oxopyrrolidin-1-yl)indole-1-carboxylate (59 mg, 68% yield) as a white solid (MS m / z 301.2 (MH + )) was given.

[0310] Step 3: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 (Ar = 2,3-dichlorophenyl) was carried out as described above in Method B for Example 28 (Step 1).

[0311] Step 4: Preparation of Example 110: 1-(1H-indol-3-yl)pyrrolidin-2-one (10.9 mg, 0.05 mmol) described in Step 2 was placed in a 4 mL vial along with the crude methyl sulfone / methyl sulfoxide mixture (51 mg, 0.09 mmol) from A-11 (Ar = 2,3-dichlorophenyl) and DMSO (1 mL). DIPEA (47 mL, 0.27 mmol) was added, and the mixture was stirred at 105 °C overnight. The mixture was cooled to room temperature, and then sodium bis(trimethylsilyl)amide (68.131 μL, 0.07 mmol) was added as a 1 M solution in THF. The resulting mixture was stirred at 105 °C for a total of 3 days. The mixture was cooled to room temperature and then quenched with 0.2 mL of glacial acetic acid. The product was isolated by preparative HPLC using a 55% to 85% methanol in water gradient (with 0.1% formic acid modifier) ​​over 15 min. After lyophilization from a mixture of water and acetonitrile, 2,3-dichloro-N-[2,4-difluoro-3-[[2-[3-(2-oxopyrrolidin-1-yl)indol-1-yl]thiazolo[5,4-d]pyrimidin-7-yl]amino]phenyl]benzenesulfonamide (Example 110; 10.7 mg, 33% yield) was obtained as a beige solid.

[0312] Preparation of Examples 111, 119 and 131: [ka]

[0313] Step 1 (for R = Me, Example 119): Potassium carbonate (0.35 g, 2.56 mmol) and 2-methoxyethanol (0.40 mL, 5.12 mmol) were added to a solution of 3-fluoro-2-nitro-aniline (0.10 g, 0.641 mmol) in DMF (3.2 mL). The resulting mixture was stirred at 80 °C for 10 h. Water was added, and the aqueous mixture was extracted with EtOAc. The organic layers were combined, washed with brine, dried over NaSO, filtered, and concentrated. The crude product was purified by column chromatography (silica gel, 0-100% EtOAc in hexane) to give 3-(2-methoxyethoxy)-2-nitro-aniline (52 mg, 38% yield). MS m / z 213.1 (MH + ).

[0314] Step 2: Preparation of substituted benzimidazoles from the 2-nitroanilines described in Step 1 was accomplished using the "one-pot" procedure described in Method G for Example 113 (Step 2).

[0315] Step 3: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 (Ar = 2,3-dichlorophenyl) was carried out as described above in Method B for Example 28 (Step 1).

[0316] Step 4: Examples 111, 119, and 131 were prepared by coupling the appropriate substituted benzimidazole from Step 2 to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using either DIPEA (Example 111) or cesium carbonate (Examples 119 and 131) in DMSO in a manner similar to that used for Example 4 (Step 7) in General Method A.

[0317] Preparation of Example 120: [ka]

[0318] Step 1: Preparation of 1H-benzo[d]imidazole-4-carbonitrile: To a nitrogen-purged (x3) 10 mL microwave vial equipped with a magnetic stir bar was added 4-bromo-1H-benzo[d]imidazole (99 mg, 0.502 mmol), dicyanozinc (70.8 mg, 0.603 mmol), palladium tetrakistriphenylphosphine (116 mg, 0.100 mmol) in DMF (4 mL). The reaction was heated to 90 °C and stirred for 16 h. The reaction was cooled to room temperature, diluted with water, and then extracted with EtOAc. The organic layer was washed with NaHCO3, brine, dried over Na2SO4, and then filtered on Celite. (登録商標) The crude material was purified by silica gel chromatography using a gradient of MeOH in DCM to give 1H-benzo[d]imidazole-4-carbonitrile (31 mg, 43% yield) as a pale pink solid. 1 H NMR (400MHz, DMSO-d6) δ:13.04(br.s.,1H),8.45(s,1H),7.90(d,J=7.83Hz,1H),7.68(d,J=7.83Hz,1H),7.20~7.44(m,1H). MS m / z 142.2 (MH + ).

[0319] Step 2: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 (Ar = 2,3-dichlorophenyl) was carried out as described above in Method B for Example 28 (Step 1).

[0320] Step 3: Examples 111, 119, and 131 were prepared by coupling 1H-benzo[d]imidazole-4-carbonitrile from Step 1 to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using DIPEA in DMSO in a manner similar to that used for Example 4 (Step 7) in General Method A.

[0321] Preparation of Example 122: [ka]

[0322] Step 1: The synthesis of 2-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]acetonitrile was described for Examples 104, 112, and 116-118. Sodium hydride (100 mg, 2.51 mmol) was added to anhydrous DMF (5 mL) in a 20 mL vial. The mixture was cooled to 0°C. 2-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]acetonitrile (240 mg, 0.84 mmol) in 5 mL of DMF was added slowly. The reaction was warmed to room temperature and then re-cooled to 0°C. 1,2-Dibromoethane (76 μL, 0.878 mmol) and sodium iodide (138 mg, 0.92 mmol) were added. The reaction was allowed to warm to room temperature and then heated to 105°C. The reaction mixture was stirred at that temperature for 24 hours. The reaction was quenched with 1M HCl at room temperature and then extracted three times with EtOAc. The combined organic layers were washed with water, then brine, dried over MgSO4, filtered, and then concentrated under reduced pressure. The crude material was purified by evaporation over Celite 400°C. (登録商標) The crude product was adsorbed onto hexanes and then purified by normal-phase silica gel chromatography using a 0% to 100% EtOAc gradient in hexanes. The desired fractions were collected and concentrated under reduced pressure to give 1-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]cyclopropanecarbonitrile (111 mg, 42% yield) as a light brown oil. MS m / z 314.2 (MH + ).

[0323] Step 2: 1-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]cyclopropanecarbonitrile (118 mg, 0.376 mmol) was dissolved in a mixture of DCM (2 mL) and TFA (2 mL) in a 20 mL vial. The reaction was stirred at room temperature overnight. The solvent was removed under reduced pressure. The material was dissolved in MeOH, and then Amberlite IRA-67 resin was added. The resin was filtered off, and then the filtrate was concentrated under reduced pressure to give 1-(1H-benzimidazol-4-yl)cyclopropanecarbonitrile; 2,2,2-trifluoroacetic acid (114 mg, 102% yield) as a light brown sticky solid. MS m / z 184.0 (MH + ).

[0324] Step 3: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 (Ar = 2,3-dichlorophenyl) was carried out as described above in the method for Example 28 (Step 1).

[0325] Step 4: Example 122 was prepared in a manner similar to that used for Example 4 (Step 7) in General Method A by coupling 1-(1H-benzimidazol-4-yl)cyclopropanecarbonitrile from Step 2; 2,2,2-trifluoroacetic acid to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using DIPEA in DMSO.

[0326] Preparation of Example 123: [ka]

[0327] Step 1: To a 100 mL round-bottom flask equipped with a magnetic stir bar was added DMF (25 mL), followed by 1H-benzo[d]imidazole-7-carboxylic acid (1.018 g, 6.28 mmol), EDCl (2.407 g, 12.56 mmol), HOBT-HO (1.923 g, 12.56 mmol), and triethylamine (1.750 mL, 12.56 mmol). The reaction was cooled to 0 °C and stirred for 2 h. Concentrated ammonium hydroxide (2 mL, 29.6 mmol) was then added, and the reaction was allowed to warm to room temperature and stirred for 24 h. The reaction was diluted with water and extracted with EtOAc. The organic layer was washed with brine, then dried over NaSO, and then concentrated under reduced pressure. The crude material was purified by filtration on Celite. (登録商標) The resulting mixture was adsorbed onto a silica gel column and purified by silica gel chromatography using a 0% to 15% ethanol in ethyl acetate gradient to give 1H-benzo[d]imidazole-7-carboxamide (281.8 mg, 1.749 mmol, 27.9% yield) as an off-white solid. MS m / z 160.1 (MH - ). 1 H NMR(400MHz,DMSO-d6) δ:12.93(br.s.,0.7H),12.41(br.s.,0.3H),9.28(br.s.,1H),8.44(br.s.,0.7H),8.16(br .s.,0.3H),7.85(d,J=5.1Hz,1H),7.76(d,J=7.4Hz,1H),7.69(br.s.,1H),7.33(br.s.,1H).

[0328] Step 2: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 (Ar = 2,3-dichlorophenyl) was carried out as described above in Method B for Example 28 (Step 1).

[0329] Step 3: Example 123 was prepared by coupling 1H-benzo[d]imidazole-7-carboxamide from Step 1 to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using DIPEA in DMSO in a manner similar to that used for Example 4 (Step 7) in General Method A.

[0330] Preparation of Example 126: [ka]

[0331] Step 1: Cesium carbonate (8.35 g, 25.6 mmol) was placed in a 50 mL flask and suspended in 6 mL of DMSO. Dimethyl malonate (2.7 mL, 30.7 mmol) was then added at room temperature. After stirring the thick slurry for 10 min, 3-fluoro-2-nitroaniline (800 mg, 5.12 mmol) was added, giving a bright orange-red mixture, which was stirred at 70 °C for 1 h. The mixture was cooled to room temperature, then diluted with EtOAc and washed with a saturated solution of ammonium chloride. The aqueous layer was extracted three times with EtOAc, and the combined organic layers were washed with brine, then dried over MgSO, filtered, and concentrated. The residue was purified by flash chromatography on silica gel using a 5% to 50% ethyl acetate in hexanes gradient. Dimethyl 2-(3-amino-2-nitrophenyl)malonate (1.225 g, 89% yield): MS m / z 269.1 (MH + ). 1 H NMR (DMSO-d6) δ:7.29(dd,J=8.2,7.4Hz,1H),7.00(dd,J=8.6,1.2Hz,1H),6.85(s,2H),6.47(dd,J=7.4,1.2Hz,1H),5.09(s,1H),3.67(s,6H).

[0332] Step 2: Dimethyl 2-(3-amino-2-nitrophenyl)malonate (1.37 g, 5.11 mmol) and lithium chloride (260 mg, 6.13 mmol) were placed in a 40 mL vial and dissolved in 6 mL of DMSO and 0.6 mL of water. The bright red mixture was heated at 150° C. for 2.5 hours, after which the color darkened slightly. The mixture was cooled to room temperature and then diluted with water and EtOAc. The biphasic mixture was filtered through a Celite (登録商標) The mixture was filtered through a short pad of hexane to remove the insoluble material that formed an emulsion, and the layers were separated. The aqueous layer was further extracted three times with EtOAc. The combined organic layers were then washed twice with water and once with brine, then dried over MgSO4, filtered, and concentrated. The bright red residue was purified by flash chromatography on silica gel using a 15% to 50% EtOAc in hexane gradient. The appropriate fractions were pooled, concentrated, and dried under reduced pressure to give methyl 2-(3-amino-2-nitro-phenyl)acetate (749 mg, 70% yield) as a bright orange solid: 1 H NMR (DMSO-d6) δ:7.24(dd,J=8.4,7.2Hz,1H),6.93(dd,J=8.4,1.4Hz,1H),6.79(s,2H),6.54(d,J=7.0Hz,1H),3.85(s,2H),3.59(s,3H).

[0333] Step 3: Methyl 2-(3-amino-2-nitrophenyl)acetate (545 mg, 2.59 mmol) was placed in a 100 mL flask equipped with a magnetic stir bar. Next, 5% palladium on charcoal (109 mg, 0.051 mmol) was added, and the mixture was suspended in 14 mL of methanol. Triethyl orthoformate (0.91 mL, 5.45 mmol) was added, followed by two drops of acetic acid. The reaction flask was placed under reduced pressure, and then hydrogen was introduced. This procedure was repeated twice more, and then the mixture was vigorously stirred at room temperature under a balloon atmosphere of hydrogen for 24 hours. 0.2 mL of Et3N was added to the mixture to neutralize the AcOH, and then it was evaporated on Celite. (登録商標) The mixture was filtered through a short pad of hexane and rinsed with methanol. The filtrate was concentrated to dryness, and the residue was then purified by flash chromatography on a silica gel column using a gradient from 1:1 EtOAc / DCM to 100% EtOAc to 10% IPA in EtOAc. The appropriate fractions were pooled, concentrated, and then dried under reduced pressure to give methyl 2-(1H-benzimidazol-4-yl)acetate (299 mg, 61% yield). MS m / z 191.2 (MH + ). 1 H NMR showed a 0.6 to 0.4 mixture of tautomers: 1 H NMR(DMSO-d6) δ:12.50(br.s.,0.4H),12.44(br.s.,0.6H),8.21(s,0.4H),8.17(s,0.6H),7.55(d,J=7.4Hz,0.4H),7.43(d,J=7.8Hz,0.6H),7. 15(t,J=7.4Hz,0.6H),7.12(t,J=7.8Hz,0.4H),7.03~7.09(m,1H),4.00(s,1.2H),3.97(s,0.8H),3.62(s,1.2H),3.60(s,1.8H).

[0334] Step 4: A solution of methyl 2-(1H-benzimidazol-4-yl)acetate (40 mg, 0.21 mmol) in 2 mL of anhydrous THF was cooled in an ice / water bath and stirred for 5 minutes. Next, a 1 M solution of lithium aluminum hydride (0.25 mL, 0.25 mmol) in THF was added dropwise. Some gas evolution was observed, and the clear yellow solution turned into a milky beige suspension. The mixture was allowed to warm to room temperature and stirred overnight. 3-4 drops of saturated ammonium chloride solution were added to the mixture. After stirring for 5 minutes, 100 mg of sodium sulfate decahydrate was added. After stirring for an additional 10 minutes, the mixture was diluted with 2-3 mL of EtOAc and evaporated on Celite. (登録商標) The filtrate was filtered through a plug of silica gel to remove insoluble material. The filtrate was concentrated to a residue, which was passed through a silica gel plug using a 30% IPA solution in EtOAc as the eluent. The filtrate was concentrated to dryness, and the resulting tan oil crystallized upon drying under reduced pressure to give 2-(1H-benzimidazol-4-yl)ethanol (34.5 mg, 100% yield) as a tan solid, which was used without further purification. MS m / z 163.0 (MH + ).

[0335] Step 5: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 (Ar = 2,3-dichlorophenyl) was carried out as described above in Method B for Example 28 (Step 1).

[0336] Step 6: Example 126 was prepared by coupling 2-(1H-benzimidazol-4-yl)ethanol from Step 4 to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using DIPEA in DMSO in a manner similar to that used for Example 4 (Step 7) in General Method A.

[0337] Preparation of Example 133: [ka]

[0338] Step 1: To a 100 mL round-bottom flask was added 4-bromo-1H-benzimidazole (1.00 g, 5.08 mmol), 2-(trimethylsilyl)ethoxymethyl chloride (1.1 mL, 6.09 mmol), and 60% NaH (244 mg, 6.09 mmol) in anhydrous DMF (10 mL). The reaction was stirred at room temperature for 1 hour. Upon completion, the reaction was quenched with saturated NH4Cl and then extracted three times with EtOAc. The organics were collected, washed with brine, separated, dried over MgSO4, filtered, and then concentrated under reduced pressure. The crude material was purified by evaporation over Celite 4000 r.p.m. (登録商標) The resulting mixture was loaded onto a 100°C column and then purified by silica gel chromatography using EtOAc in hexanes to give 2-[(4-bromobenzimidazol-1-yl)methoxy]ethyl-trimethyl-silane (1.18 g, 71% yield) as a brown oil. MS m / z 327.2 (MH + ).

[0339] Step 2: To a flame-dried 5 mL microwave vial, 2-[(4-bromobenzimidazol-1-yl)methoxy]ethyl-trimethyl-silane (500 mg, 1.53 mmol) and bis(pinacolato)diboron (776 mg, 3.06 mmol) were added in DMF (0.30 mL) under a N atmosphere. Next, 1,1'-bis(diphenylphosphino)ferrocene-palladium(II) dichloride dichloromethane complex (279 mg, 0.382 mmol) and potassium acetate (450 mg, 4.58 mmol) were added to the solution. The reaction was sparged with a balloon of argon for several minutes, sealed, and then heated to 100 °C and stirred for 16 h. The reaction was cooled to room temperature. The crude reaction mixture was used directly in the next step.

[0340] Step 3: To the crude reaction mixture from Step 2 was added 4-amino-3-bromopyridine (34 mg, 0.20 mmol), tetrakis(triphenylphosphine)palladium(0) (35 mg, 0.0305 mmol), and sodium carbonate (97 mg, 0.914 mmol). The reaction was sparged with a balloon of argon for about 5 to about 10 minutes, sealed, and then heated at 100° C. for 24 hours. Upon completion, the reaction was cooled to room temperature. The reaction was diluted with brine and extracted three times with EtOAc. The organic layers were combined, dried over MgSO4, filtered, and then concentrated under reduced pressure. The crude material was purified by filtration on Celite. (登録商標) The resulting mixture was loaded onto a SiO2 column and then purified using MeOH in DCM to give 3-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]pyridin-4-amine (72 mg, 69% yield) as a brown film. MS m / z 341.2 (MH + ).

[0341] Step 4: To a 20 mL scintillation vial was added 3-[1-(2-trimethylsilylethoxymethyl)-benzimidazol-4-yl]pyridin-4-amine (72 mg, 0.210 mmol) in DCM (2 mL) and TFA (2 mL). The reaction was stirred at room temperature for 16 hours. Upon completion, the solvent was removed under reduced pressure and then dried under vacuum to give 3-(1H-benzimidazol-4-yl)pyridin-4-amine TFA salt (68 mg, 100% yield) as a brown oil. MS m / z 211.2 (MH + ).

[0342] Step 5: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 (Ar = 2,3-dichlorophenyl) was carried out as described above in Method B for Example 28 (Step 1).

[0343] Step 6: Example 133 was prepared by coupling 3-(1H-benzimidazol-4-yl)pyridin-4-amine TFA salt from Step 4 to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using DIPEA in DMSO in a manner similar to that used for Example 4 (Step 7) in General Method A.

[0344] Preparation of Example 134: [ka]

[0345] Step 1: To a 20 mL scintillation vial was added 4-bromo-1H-benzimidazole (500 mg, 2.54 mmol), 60% NaH (122 mg, 3.05 mmol), and 4-methoxybenzyl chloride (413 μL, 3.05 mmol) in DMF (5 mL). The reaction was stirred at room temperature for 2 hours. The reaction was quenched with saturated NH4Cl and then extracted three times with EtOAc. The organics were collected, dried over MgSO4, filtered, and then concentrated under reduced pressure. The crude material was purified by filtration using Celite. (登録商標) The oil was loaded onto a 100° C. column and then purified by silica gel chromatography using EtOAc in hexanes to give 4-bromo-1-[(4-methoxyphenyl)methyl]benzimidazole (735 mg, 91% yield) as a brown oil. 1 H NMR(400MHz,DMSO-d6) δ:8.50(s,1H),7.57(dd,J=8.2,0.8Hz,1H),7.29(ddd,J=8.6,3.1,2.0Hz ,2H),7.14(t,J=7.8Hz,2H),6.86~6.89(m,2H),5.43(s,2H),3.70(s,3H). Minor isomers: 8.46(s,1H),7.70(dd,J=8.0,1.0Hz,1H),7.42(d,J=7.8Hz,2H),7.04(d,J=9.0Hz,2H),6.90(m,J=3.5Hz,2H),5.72(s,2H),3.70(s,3H). MS m / z 317.2(MH+ ).

[0346] Step 2: To a flame-dried 5 mL microwave vial, 4-bromo-1-[(4-methoxyphenyl)methyl]benzimidazole (200 mg, 0.63 mmol) from Step 1 and bis(pinacolato)diboron (320 mg, 1.26 mmol) were added in DMF (0.30 mL) under a N atmosphere. Next, 1,1'-bis(diphenylphosphino)ferrocene-palladium(II) dichloride dichloromethane complex (231 mg, 0.315 mmol) and then potassium acetate (186 mg, 1.89 mmol) were added to the solution. The reaction was sparged with a balloon of argon for several minutes, sealed, and then heated to 100 °C and stirred for 24 h. The reaction was cooled to room temperature. The reaction was extracted, diluted with brine, and extracted three times with EtOAc. The organics were collected, dried over MgSO4, and filtered through Celite (登録商標) The crude material was filtered through a plug of Celite and then concentrated under reduced pressure. (登録商標) The resulting mixture was loaded onto a silica gel column and then purified by silica gel chromatography using EtOAc in hexanes. The desired fractions were collected and concentrated under reduced pressure to give 1-[(4-methoxyphenyl)methyl]-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzimidazole (82 mg, 36% yield) as a brown oil. MS m / z 365.4 (MH + ).

[0347] Step 3: To a flame-dried 5 mL microwave vial under a N atmosphere, 1-[(4-methoxyphenyl)methyl]-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzimidazole (56 mg, 0.154 mmol) and 2-amino-3-bromopyridine (35 mg, 0.200 mmol) were added. Next, sodium carbonate (49 mg, 0.461 mmol) and then tetrakis(triphenylphosphine)palladium(0) (18 mg, 0.0154 mmol) were added to the solution. The reaction was sparged with a balloon of argon for about 5 to about 10 minutes, sealed, and then heated to 80 °C and stirred for 16 hours. The reaction was cooled to room temperature. The reaction was diluted with brine and extracted three times with EtOAc. The organics were combined, dried over MgSO4, filtered, and then concentrated under reduced pressure. The crude material was purified by evaporation over Celite (登録商標) The resulting mixture was loaded onto a silica gel column and then purified by silica gel chromatography using MeOH in DCM. The desired fractions were collected and concentrated under reduced pressure to give 3-[1-[(4-methoxyphenyl)methyl]benzimidazol-4-yl]pyridin-2-amine (15 mg, 0.0445 mmol, 29% yield) as a white solid. MS m / z 331.2 (MH + ).

[0348] Step 4: To a 20 mL scintillation vial was added 3-[1-[(4-methoxyphenyl)methyl]-benzimidazol-4-yl]pyridin-2-amine (15 mg, 0.0445 mmol) in TFA (2 mL). The reaction was heated to 80° C. and stirred for 96 hours. The solvent was removed under reduced pressure, and the product was then co-evaporated with toluene three times to give 3-(1H-benzimidazol-4-yl)pyridin-2-amine TFA salt (14 mg, 100%). MS m / z 211.2 (MH + ).

[0349] Step 5: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 (Ar = 2,3-dichlorophenyl) was carried out as described above in Method B for Example 28 (Step 1).

[0350] Step 6: Example 134 was prepared in a manner similar to that used for Example 4 (Step 7) in General Method A by coupling 3-(1H-benzimidazol-4-yl)pyridin-2-amine TFA salt from Step 4 to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using DIPEA in DMSO.

[0351] Preparation of Example 143: [ka]

[0352] Step 1: Methyl 2-(1H-benzimidazol-4-yl)acetate (303 mg, 1.59 mmol), prepared as described in Step 3 of Example 126, was dissolved in 10 mL of DMF, and then potassium carbonate (661 mg, 4.78 mmol) was added, followed by the dropwise addition of SEM-Cl (0.42 mL, 2.39 mmol) at room temperature over 10 minutes. The mixture was stirred at the same temperature for 16 hours. The mixture was poured into a saturated solution of ammonium chloride and then extracted three times with EtOAc. The combined organic layers were washed with water, then brine, dried over MgSO4, filtered, and concentrated. The resulting residue was purified by flash chromatography on silica gel using a gradient of 20% ethyl acetate in hexanes to 100% EtOAc. The appropriate fractions were pooled, concentrated, and dried under reduced pressure. A mixture of methyl 2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-4-yl)acetate and methyl 2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-7-yl)acetate (260 mg, 51% yield) was obtained as a pale yellow oil.

[0353] Step 2: A mixture of 2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-4-yl)methyl acetate and 2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-7-yl)methyl acetate (265 mg, 0.83 mmol) was placed in a 25 mL flask. A solution of diphenyl(vinyl)sulfonium trifluoromethanesulfonate (479 mg, 1.32 mmol) in DMSO (4 mL) was added, followed by 1,8-diazabicyclo[5.4.0]undec-7-ene (0.37 mL, 2.48 mmol) at room temperature. The mixture was stirred at that temperature for 17 hours. The reaction mixture was then diluted with water containing a small amount of NH4Cl and EtOAc. The layers were separated, and the aqueous layer was further extracted twice with EtOAc. The combined organic layers were washed twice with water and once with brine. The organic layer was then dried over MgSO, filtered, and concentrated. The resulting residue was purified by flash chromatography on silica gel using a 20% to 100% EtOAc gradient in hexanes, followed by 5% IPA in EtOAc. The appropriate fractions were pooled, concentrated, and dried under reduced pressure to give methyl 1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-4-yl)cyclopropane-1-carboxylate and methyl 1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-7-yl)cyclopropane-1-carboxylate as a pale yellow oil as an inseparable mixture of isomers (250 mg, 87% yield). MS m / z 347.2 (MH + ).

[0354] Step 3: A solution of the methyl ester from step 2 (275 mg, 0.79 mmol) in 4 mL of anhydrous THF was cooled in an ice / water bath and stirred for 5 minutes. Next, a 1 M solution of lithium aluminum hydride (0.95 mL, 0.95 mmol) in THF was added dropwise. Some gas evolution was observed. The mixture was allowed to warm to room temperature and stirred for 23 hours. Sodium sulfate decahydrate (0.7 g) was added to the mixture, which was further stirred for 1 hour. The mixture was then diluted with EtOAc and evaporated on Celite. (登録商標) The mixture was filtered through a pad of hexane and washed with EtOAc. The filtrate was then concentrated to dryness, and the residue was purified by flash chromatography on silica gel using a gradient from 100% EtOAc to 30% IPA in EtOAc.

[0355] The first eluting product was [1-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]cyclopropyl]methanol (132 mg, 52% yield). MS m / z 319.2 (MH + ). 1 H NMR(DMSO-d6) δ:8.33(s,1H),7.48(d,J=8.2Hz,1H),7.18(t,J=7.8Hz,1H),7.10(d,J=7.4Hz,1H),5.61(s,2H),4.98(t,J=5.7H) z,1H),3.70(d,J=5.5Hz,2H),3.50(t,J=8.0Hz,2H),0.91(d,J=2.7Hz,4H),0.83(t,J=8.0Hz,2H),-0.08(s,9H).

[0356] The second eluting product was [1-(1H-benzimidazol-4-yl)cyclopropyl]methanol (47 mg, 31% yield). MS m / z 186.9 (MH - ).

[0357] Step 4: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 (Ar = 2,3-dichlorophenyl) was carried out as described above in Method B for Example 28 (Step 1).

[0358] Step 5: Example 143 was prepared by coupling [1-(1H-benzimidazol-4-yl)cyclopropyl]methanol from Step 3 to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using DIPEA in DMSO in a manner similar to that used for Example 4 (Step 7) in General Method A.

[0359] Preparation of Example 154: [ka]

[0360] Step 1: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 (Ar = 2-methyl-3-fluorophenyl) was carried out as described above in Method B for Example 28 (Step 1).

[0361] Step 2: The methyl sulfone / methyl sulfoxide mixture from Step 1 (82 mg, 0.16 mmol) was suspended in 1.3 mL of 95% EtOH and 0.1 mL of water. Next, a 1 M solution of hydrazine (0.77 mL, 0.77 mmol) in THF was added. The resulting mixture was stirred at 45° C. for 2 hours. The mixture, which became a pale beige suspension, was cooled to room temperature and then diluted with 10 mL of water. The solid from the resulting suspension was collected on a hardened paper filter in a Buchner funnel (very slow filtration). The solid was washed with a small amount of water and then dried under reduced pressure. N-[2,4-difluoro-3-[(2-hydrazinothiazolo[5,4-d]pyrimidin-7-yl)amino]phenyl]-3-fluoro-2-methyl-benzenesulfonamide (71 mg, 95% yield) was obtained as an off-white solid. MS m / z 482.2 (MH + ). 1 H NMR(400MHz,DMSO-d6) δ:10.38(br.s.,1H),9.21(s,1H),8.79(s,1H),7.98(s,1H),7.59(d,J=7.8Hz,1H),7.47(t,J=8.6Hz,1H),7.38 (td,J=7.9,5.7Hz,1H),7.17(td,J=8.9,5.9Hz,1H),7.10(t,J=9.0Hz,1H),5.20(s,2H),2.47(d,J=2.1Hz,3H).

[0362] Step 3: Isopentyl nitrite (0.038 mL, 0.28 mmol) was placed in a 25 mL flask and then diluted with 4 mL of acetonitrile. Copper dibromide (50.7 mg, 0.23 mmol) was then added, and the dark green suspension was degassed by bubbling argon through it for 5 minutes and then ultrasonically degassed for another 1 minute. Next, N-[2,4-difluoro-3-[(2-hydrazinothiazolo[5,4-d]pyrimidin-7-yl)amino]phenyl]-3-fluoro-2-methyl-benzenesulfonamide (91 mg, 0.19 mmol) (from Step 2) was added in four portions at room temperature (some gas evolution was observed). Once the addition was complete, the mixture was stirred for 20 minutes. It was then diluted with EtOAc and washed with water containing 3-4 mL of saturated aqueous EDTA solution. After separation of the layers, the wash was repeated two more times (until the blue color disappeared in the aqueous layer). The organic layer was then washed with water and then brine. It was dried over MgSO4, filtered, and concentrated. The residue was purified by silica gel chromatography using a gradient of 80% DCM, 20% hexane to 100% DCM, followed by a gradient of 45% EtOAc in DCM. The appropriate fractions were pooled, concentrated, and dried to give N-[3-[(2-bromothiazolo[5,4-d]pyrimidin-7-yl)amino]-2,4-difluoro-phenyl]-3-fluoro-2-methyl-benzenesulfonamide (80 mg, 80% yield) as a white solid.

[0363] Step 4: Preparation of Example 154: N-[3-[(2-bromothiazolo[5,4-d]pyrimidin-7-yl)amino]-2,4-difluoro-phenyl]-3-fluoro-2-methyl-benzenesulfonamide (36 mg, 0.07 mmol) from Step 3 was placed in a 10 mL flask along with tributyl(thiazol-5-yl)stannane (38 mg, 0.10 mmol) and a magnetic stir bar. DMF (2 mL) was added, and argon was bubbled through the mixture for 3-4 minutes. Tetrakis(triphenylphosphine)palladium(0) (7.8 mg, 0.007 mmol) was added. Argon was bubbled through the mixture ultrasonically for an additional 3-4 minutes. The yellow solution was immersed in an oil bath preheated to 110 °C. After stirring at that temperature for 6 hours, the reaction was cooled to room temperature and acidified with a few drops of formic acid. It was then concentrated to an oil, which was diluted to 2 mL with 1:1 DMSO and methanol. The solution was filtered through a syringe filter and purified by reverse-phase preparative HPLC (two injections; 50% to 100% MeOH gradient in water, 0.1% formic acid). Appropriate fractions were pooled and concentrated. The product was lyophilized from a mixture of acetonitrile and water to give N-[2,4-difluoro-3-[(2-thiazol-5-ylthiazolo[5,4-d]pyrimidin-7-yl)amino]phenyl]-3-fluoro-2-methyl-benzenesulfonamide (12.7 mg, 35% yield) as an off-white solid.

[0364] Preparation of Example 156: [ka]

[0365] Step 1: Preparation of Example 156: The precursor, tert-butyl 4-(1-(7-((3-((2,3-dichlorophenyl)sulfonamido)-2,6-difluorophenyl)amino)thiazolo[5,4-d]pyrimidin-2-yl)-1H-benzo[d]imidazol-4-yl)piperazine-1-carboxylate, was prepared as described in general procedure G for Example 113 using Boc-protected piperazine for the first step. This precursor (63 mg, 0.08 mmol) was dissolved in 0.8 mL of DCM, and TFA (0.061 mL, 0.80 mmol) was added at room temperature. The mixture was stirred for 20 hours, and then 100 mg of solid cesium carbonate was added. After stirring for several minutes, the solid was filtered off, and the filtrate was concentrated under reduced pressure to a residue that was purified by reverse-phase preparative HPLC (two injections, 50% MeOH in water to 100% MeOH gradient, 0.1% formic acid buffer) to give 2,3-dichloro-N-[2,4-difluoro-3-[[2-(4-piperazin-1-ylbenzimidazol-1-yl)thiazolo[5,4-d]pyrimidin-7-yl]amino]phenyl]benzenesulfonamide (47 mg, 85% yield) (Example 156) as an off-white solid after lyophilization from an acetonitrile / water mixture.

[0366] Preparation of Example 158: [ka]

[0367] Step 1: The starting material, 4-thiomorpholine benzimidazole fragment, was prepared according to Method G. oxone in water (10 mL) (登録商標)A solution of (1.42 g, 2.32 mmol) was added to a solution of 4-(1H-benzimidazol-4-yl)thiomorpholine (0.127 g, 0.579 mmol) in methanol (20 mL). The resulting mixture was stirred at room temperature for 17 h. The mixture was concentrated in vacuo, and the crude product was purified by column chromatography (silica gel, 0-25% MeOH in DCM) to give 4-(1H-benzimidazol-4-yl)-1,4-thiazinane 1,1-dioxide (0.126 g, 87% yield) as a solid. MS m / z 252.2 (MH + ).

[0368] Step 2: Preparation of a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 (Ar = 2,3-dichlorophenyl) was carried out as described above in Method B for Example 28 (Step 1).

[0369] Step 3: Example 158 was prepared by coupling 4-(1H-benzimidazol-4-yl)-1,4-thiazinane 1,1-dioxide from Step 1 to the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar=2,3-dichlorophenyl) using cesium carbonate in DMSO in a manner similar to that used for Example 4 (Step 7) in General Method A.

[0370] Preparation of Example 159: [ka]

[0371] Step 1: Carbomethoxylation of 2,4,5-trifluoroaniline was carried out as described in patent application WO2020 / 261156.

[0372] Step 2: To a solution of methyl 3-amino-2,5,6-trifluorobenzoate (2.72 g, 13.26 mmol) in DCE / pyridine (1:1, 16 mL) at room temperature, 2,3-dichlorobenzenesulfonyl chloride (3.91 g, 15.91 mmol) was added portionwise. The reaction was heated at 70 °C for 16 h. The reaction was monitored by LCMS. Upon completion, it was quenched with 1 M HCl. The aqueous layer was extracted three times with EtOAc (15 mL). The combined organic layers were washed with brine, dried over MgSO4, and evaporated to dryness. The residue was purified by chromatography on a silica gel column using 0 to 30% EtOAc in hexanes. Pure fractions were collected and evaporated to give methyl 3-((2,3-dichlorophenyl)sulfonamido)-2,5,6-trifluorobenzoate (5.14 g, 91% yield) as a light brown solid: MS m / z 412.0 (MH + ).

[0373] Step 3: To a solution of methyl 3-((2,3-dichlorophenyl)sulfonamido)-2,5,6-trifluorobenzoate (5.14 g, 12.41 mmol) from Step 2 in 30 mL of THF:MeOH (5:1) was added 2 M KOH (37 mL, 74.5 mmol) at room temperature. The reaction was stirred overnight at room temperature. Upon completion, it was evaporated to dryness, and to the residue was added water (30 mL) and diethyl ether (30 mL). The aqueous layer was washed twice with ether (20 mL). The aqueous layer was acidified with 1 M hydrochloric acid to pH = 2. The aqueous layer was extracted three times with EtOAc (30 mL). The combined organic layers were washed with brine, dried over MgSO, filtered, and evaporated to give 3-((2,3-dichlorophenyl)sulfonamido)-2,5,6-trifluorobenzoic acid (4.5 g, 91% yield) as a pale orange oil. The compound was used directly in the next step. MS m / z 398.0 (MH + ).

[0374] Step 4: To a solution of 3-((2,3-dichlorophenyl)sulfonamido)-2,5,6-trifluorobenzoic acid (4.50 g, 11.2 mmol) from Step 3 in acetonitrile (30 mL) was added triethylamine (1.71 mL, 12.37 mmol) and diphenylphosphoryl azide (2.91 mL, 13.50 mmol) at room temperature. The reaction was heated at 80 °C overnight. The reaction was cooled to room temperature, and water (30 mL) was added. The aqueous layer was extracted three times with EtOAc (30 mL). The combined organic layers were washed with brine, dried over MgSO, filtered, and evaporated to a dark yellow residue. The crude material was purified by chromatography on a silica gel column using 0 to 50% EtOAc in hexanes. The pure fraction was collected and evaporated to give 2,3-dichloro-N-(2,4,5-trifluoro-3-isocyanatophenyl)benzenesulfonamide (2.29 g, 51% yield) as a brown solid. MS m / z 391.0 (MH + ).

[0375] Step 5: To a solution of 2,3-dichloro-N-(2,4,5-trifluoro-3-isocyanatophenyl)benzenesulfonamide (1.35 g, 3.39 mmol) from Step 4 in THF (17 mL) was added 4 M aqueous LiOH (17 mL). The pressure vessel was sealed and heated in an oil bath at 100 °C for 1 h. When the reaction was complete, it was quenched with saturated NH4Cl, EtOAc was added, and the layers were separated. The aqueous layer was extracted twice with EtOAc (20 mL). The combined organic layers were washed with brine, dried over MgSO4, filtered, and evaporated to give N-(3-amino-2,4,5-trifluorophenyl)-2,3-dichlorobenzenesulfonamide (1.09 g, 86% yield) as a brown solid. The compound was carried on to the next step without further purification: 1H NMR(400MHz,DMSO-d6) δ:10.59(br.s.,1H),7.94(dd,J=8.2,1.6Hz,1H),7.89(dd,J=8.2,1.6Hz,1H),7.51(dd,J=8.0Hz,1H),6.34~6.43(m,1H),5.72(s,2H). MS m / z=369.0.

[0376] Step 6: N-(3-amino-2,4,5-trifluorophenyl)-2,3-dichloro-benzenesulfonamide (300 mg, 0.81 mmol) from Step 5 and 7-chloro-2-methylsulfanyl-thiazolo[5,4-d]pyrimidine (A-10, 194 mg, 0.89 mmol) were added to glacial AcOH (3.2 mL). The reaction was heated at 65° C. overnight and at 85° C. for 2 hours. The remainder of 7-chloro-2-methylsulfanyl-thiazolo[5,4-d]pyrimidine (A-10, 100 mg, 0.46 mmol) was added, and the mixture was stirred at 100° C. for 24 hours. It was then cooled to room temperature and concentrated under reduced pressure to a residue. Water and DCM were added, and the layers were separated. The aqueous layer was further extracted with DCM. The combined organic layers were washed with brine, dried over MgSO, filtered, and concentrated under reduced pressure to a brown solid, which was carried on to the next reaction without further purification. MS m / z 552.0 (MH + ).

[0377] Step 7: To the crude 2,3-dichloro-N-[2,4,5-trifluoro-3-[(2-methylsulfanylthiazolo[5,4-d]pyrimidin-7-yl)amino]phenyl]benzenesulfonamide (238 mg, 0.43 mmol) from Step 6 in DCM (5 mL) was added 3-chloroperoxybenzoic acid (149 mg, 0.862 mmol). The resulting reaction mixture was stirred overnight at room temperature, then diluted with EtOAc and washed with a saturated aqueous solution of NaHCO3, followed by brine. The organic layer was dried over MgSO4, filtered, and concentrated. The resulting crude product (mostly methyl sulfone, MS m / z 584.2 (MH +)) was carried on to the next step without further purification (66 mg, 26% yield).

[0378] Step 8: Crude 2,3-dichloro-N-[2,4,5-trifluoro-3-[(2-methylsulfonylthiazolo[5,4-d]pyrimidin-7-yl)amino]phenyl]benzenesulfonamide (66 mg, 0.11 mmol) was dissolved in 1.1 mL of DMSO. Benzimidazole (13 mg, 0.11 mmol) and cesium carbonate (74 mg, 0.23 mmol) were added, and the reaction mixture was stirred at 100° C. overnight. After cooling, the reaction mixture was separated by silica gel and Celite (登録商標) The filtrate was filtered through a short pad of hexane. The filtrate was purified by reverse-phase chromatography using a methanol-in-water gradient with 0.1% formic acid as a modifier. The appropriate fractions were pooled and concentrated. The residue was lyophilized from an acetonitrile / water mixture to give N-[3-[[2-(benzimidazol-1-yl)thiazolo[5,4-d]pyrimidin-7-yl]amino]-2,4,5-trifluoro-phenyl]-2,3-dichloro-benzenesulfonamide (Example 159, 12 mg, 17% yield) as a fluffy white powder.

[0379] biological activity

[0380] (a) Kinase activity assays for BRAF, CRAF, and ARAF

[0381] Compound preparation: Solid samples of each substance in 1 dram vials were suspended in DMSO (Fisher Scientific) at a stock concentration of 20 mM. Stocks were stored at -20°C and protected from light. If compound solubility at 20 mM was deemed problematic, the initial concentration of the DMSO stock was changed to 10 mM or 5 mM.

[0382] In vitro enzyme reactions were used to assess compound-specific activity against BRAF, CRAF, and ARAF. For BRAF and CRAF, 0.375 nM purified GST-tagged kinase (Millipore Sigma, catalog no. B4062-10UG and catalog no. R1656-10UG, respectively) was incubated with 75 nM kinase-dead MEK1 substrate (catalog no. 40075; BPS Bioscience) in the presence of 10 μM Ultrapure ATP (catalog no. V9102; Promega; part V915A) in a buffer containing 50 mM HEPES (pH 7.5), 10 mM MgCl2, 1 mM EDTA, 0.01% Brij-35, and 2 mM DTT, with or without test compound. Separate reactions were performed using the MEK1 substrate and ATP as a blank control. The ARAF kinase reaction was exactly the same except the kinase concentration was increased to 3.75 nM (catalog no. 1768-0000-1; Reaction Biology).

[0383] For compound treatment, 5 μL / well of the test substance solution was placed in a 384-well proxy plate (Perkin Elmer) and mixed with 2x concentrated kinase reaction solution. A dilution series was selected so that nine concentrations covered the range from 100 nM to 0.01 nM. If necessary (if the compound showed low specific potency), the initial concentration of 100 nM was changed to 1 μM or 0.5 μM, and further dilutions were made accordingly. The final concentration of DMSO in the assay was set to 0.05%.

[0384] BRAF and CRAF kinase reactions were carried out at 30°C for a total of 2 hours and then stopped by diluting the reaction mixture in half with ADP-Glo ​​Reagent (Cat. No. V9102; Promega; part V912C). The reactions were then incubated at room temperature for 1 hour, after which 1 volume of Kinase Detection Reagent (Cat. No. V9102; Promega; part V917A) was added. The plate was then equilibrated at room temperature for 30 minutes before detecting luminescence with a Synergy Neo2 plate reader (Biotek). The effect of each compound dilution on BRAF and CRAF kinase activity was expressed as % inhibition and calculated as follows: First, the internal 100% inhibition control (the average luminescence in kinase reactions containing only kinase-dead MEK1 substrate) was subtracted from each data point. The average of the DMSO (vehicle) controls (set as 0% inhibition) was set, and the following formula was used to calculate % inhibition:

number

[0385] The ARAF kinase reaction was carried out for a total of 2 hours at 30°C and then stopped by adding EDTA to a final concentration of 40 mM. (登録商標) SureFire (登録商標) Ultra 商標 Detection was performed using the p-MEK 1 / 2 (Ser218 / 222) (Perkin Elmer) kit. Reactions were performed in 384-well proxy plates (Perkin Elmer) with 5 μL of kinase according to the manufacturer's specifications, followed by AlphaLISA. (登録商標) The reaction was incubated overnight at room temperature in a humidified chamber. (登録商標)Signals were recorded on a Synergy Neo2 plate reader (Biotek) equipped with a filter. The effect of each compound dilution on the pMEK signal generated by the ARAF reaction was expressed as % inhibition and calculated using the following formula: To measure background pMEK, an internal 100% inhibition control (average luminescence in kinase reactions containing only kinase-dead MEK1 substrate) was included in each plate and subtracted from each data point. The average of the DMSO (vehicle) controls (set as 0% inhibition) was also set and used to calculate % inhibition.

number

[0386] I C 50 Values ​​were obtained by plotting kinase inhibition values ​​and fitting dose-activity curves with a log(agonist) vs. response-variable slope (four parameter) function using GraphPad Prism (v7.0) or the Dotmatics Screening Ultra platform. Standards included in the ARAF kinase assay were Belvarafenib (MedChem Express, catalog number HY-109080; CAS number 1446113-23-0), LXH254 (MedChem Express, catalog number HY-112089; CAS number 1800398-38-2), and BGB283 (catalog number HY-18957; CAS number 1446090-79-4).

[0387] Therefore, all substances reported herein are ATP-competitive kinase inhibitors of BRAF, CRAF, and ARAF, as demonstrated by direct inhibition of enzymatic activity in vitro. The BRAF and CRAF inhibitory potencies of the compounds are listed in Tables 3 and 4 below, while the ARAF kinase inhibitory potencies of representative analogs are listed in Table A below.

[0388] [Table A]

[0389] For the ARAF biochemical kinase assay, * denotes IC 50 indicates IC >10 nM, ** indicates IC between 1 and 10 nM 50 indicates the range of IC 50 <1 nM.

[0390] (b) General cell culture methods

[0391] All cancer cell lines (A375, A101D, A2058, RKO, HT29 SK-MEL 30, IPC298, HepG2, HCT-116, Lovo, SW620, SW480, NCI-H358, NCI-H2122, Calu-6, NCIH2087, NCIH1755, NCIH1666, and Mewo) were obtained from ATCC and cultured in RPMI-1640 medium (Gibco) supplemented with 5% heat-inactivated fetal bovine serum (FBS, Wisent) at 37°C and 5% CO2. Cells were maintained in T175 flasks (Greiner). The culture medium was removed, washed once with 10 mL of room temperature phosphate-buffered saline (PBS; Wisent), and passaged by incubation with 2 mL of 0.05% trypsin (Thermo-Fisher) at 37°C. The trypsin was then inactivated by adding complete growth medium, and the cells were replated in T175 culture dishes at the appropriate dilution. All cell lines were routinely tested for mycoplasma contamination. The tissue type and mutation status of each cell line can be seen in Table B below.

[0392] [Table B]

[0393] (c)AlphaLISA (登録商標) SureFire (登録商標) Ultra 商標 Measurement of phospho-ERK inhibition by p-ERK 1 / 2 (Thr202 / Tyr204) in cultured human cancer cell lines

[0394] AlphaLISA (登録商標) SureFire (登録商標) Ultra 商標 p-ERK 1 / 2 (Thr202 / Tyr204) analysis was performed on cells seeded in 96-well flat-bottom clear dishes (Costar) in 100 μL of complete RPMI-1640 growth medium at the densities indicated in Table C below. Cells were maintained overnight at 37°C under 5% CO2 before being treated with serial dilutions of compounds for 1 hour. Cell density (cells / cm)2 ) is the number of cells per well of a 96-well plate (0.143 cm 2 ) is equivalent to dividing it by

[0395] [Table C]

[0396] In the dilution series, 100 μL / well of test substance dilutions prepared in complete RPMI-1640 growth medium were added to the cells. The dilution series was selected to cover the range of 10 μM to 0.33 nM with 10 concentrations. If necessary (as in the case of A375 and H1666 cells, which are generally more sensitive to compounds), the initial concentration of 10 μM was increased to 100 μM or decreased to 1 μM, and further dilutions were made accordingly. The final concentration of DMSO in the assay was set to 0.5%.

[0397] After treatment, the medium was removed and the cells were lysed in 50 μL of 1× AlphaScreen Ultra Lysis Buffer (Perkin Elmer). (登録商標) SureFire (登録商標) Ultra 商標 p-ERK 1 / 2 (Thr202 / Tyr204) (Perkin Elmer) reactions were performed in 384-well proxy plates (Perkin Elmer) according to the manufacturer's specifications using 5 μL of cell lysate, followed by incubation of the reaction overnight at room temperature in a humidified chamber. After completion of the reaction, the reaction was analyzed by the built-in AlphaLISA on an EnVision plate reader (Perkin Elmer). (登録商標) The signal was recorded using the settings.

[0398] The effect of each compound dilution on the pERK signal was expressed as % inhibition and calculated using the following formula: An internal 100% inhibition control (1 mM trametinib, catalog number HY-10999; MedChem Express; CAS number 871700-17-3) was included in each plate and used as a measure of pERK background. First, the value obtained for trametinib was subtracted from each data point. The mean value of the DMSO (vehicle) control (set as 0% inhibition) was set and used to calculate % inhibition.

number

[0399] The ability of each compound to inhibit pERK signaling was determined by plotting the inhibition value for each data point in the dilution series and fitting the resulting curve with a log(agonist) vs. response-variable slope (four parameter) function using GraphPad Prism (V7.0) or the Dotmatics Screening Ultra platform. 50 expressed as a value.

[0400] If present, paradoxical pERK induction is determined by the pERK IC of the compound. 50 The % inhibition of the lowest data point in the dose-activity curve was estimated from the negative % inhibition values ​​observed in the curve. 最小 %) was set to less than -20%, which is considered within the expected assay variation (e.g., Y 最小 Compounds with % = -30%, -50%, or -150% are considered to produce paradoxical induction of the pathway, while compounds with Y 最小 =-10% IC 50 (Compounds that exhibit a curve are considered not to cause paradoxical activation of the pathway.) Therefore, compounds that meet the following criteria in a given cell line are said to inhibit the pathway without causing paradoxical induction: 1. The percentage of inhibition at the highest dose (30 μM, 10 μM or 1 μM) exceeded 50%. 2.IC 50 Station line Y 最小 % was -20% or more. 最小 is the IC of the compound 50 It corresponds to the data point with the lowest value on the curve.

[0401] It is well known to those skilled in the art that some variability in inhibition values ​​is to be expected in such experiments. 最小 Values ​​of ±20% are considered within experimental error and not significant. Therefore, only compounds with negative values ​​(greater than about 20%) beyond assay variability are considered to induce paradoxical activation of signaling cascades and are not within the scope of this disclosure. Figure 1 shows the IC values ​​of a compound that induces paradoxical pathway activation (PLX4720, commercially available from Selleck Chemical; CAS No. 918505-84-7) and a representative compound described herein that exhibits an unexpected and distinct induction-free profile. 50 Provide a visualization of the curve.

[0402] Figure 1 shows that RAS mutant HCT116 cells (Examples 44 and 122) do not induce paradoxical induction of pERK signaling (Y 最小 >-20%) compared with the compounds described herein, which showed strong induction of the pathway (Y 最小 Representative IC for compound (PLX4720) causing cytotoxicity of 2000-2000% (~-600%) 50 Inhibitory dose-response curves are shown.

[0403] Importantly, the compounds do not induce paradoxical activation of the pathway according to the criteria set out above.

[0404] The compounds of the examples 1-159 exhibit pERK inhibitory activity in the colon G13D Ras mutated HCT-116 cell line, as shown in Tables 3 and 4. In addition, some examples were also shown to exhibit inhibition without paradoxical induction of pERK signaling in the SW480 colon cancer cell line harboring the G12D allele of KRAS (Tables 3 and 4). Moreover, some examples in Tables 3 and 4 exhibit pERK inhibitory activity in the BRAF V600E Driver mutation (BRAF V600E The compounds were also tested for inhibition of pERK in A375 cells containing a pERK driver mutation and were found to be similarly active (Tables D-1 and D-2). All compounds, Examples 1 to 159, had pERK IC values ​​of less than 30 μM in the HCT116 cell line. 50 The values ​​were shown.

[0405] Representative compounds defined herein were also tested for their pERK inhibitory activity against additional tumor cells and showed good to very good pERK inhibitory activity in cancer cell lines harboring various NRAS, KRAS, and NF1 alleles and representing a wide variety of tissue types (i.e., SK-MEL 30, IPC298, HepG2, HCT-116, Lovo, SW620, SW480, NCI-H358, NCI-H2122, Calu-6, and Mewo; see Tables D-1 and D-2 below, and Table B for genotypes). The pERK inhibitory activity of the compounds was stronger in cancer cell lines harboring various BRAF alleles (A375, A101D, A2058, RKO, HT29, NCIH2087, NCIH1755, and NCIH1666) (Tables D-1 and D-2).

[0406] Table D (Table D-1 and Table D-2). Panel of RAS-mutated cancer cell lines (see Table B for genotypes) and BRAF V600E No inducible pERK IC for selected compounds in mutant A375 50 values ​​and antiproliferative EC 50 value.

[0407] [Table D-1]

[0408] In pERK, + is IC 50 >0.3 μM, and ++ indicates IC of 0.03–0.3 μM. 50 indicates the range of IC 50 <0.03 μM. For proliferation, * indicates EC 50 indicates proliferation >3 μM, ** indicates EC 50 indicates the range, and *** indicates EC 50 Indicates <0.3 μM. Belv: Belvarafenib. Values ​​in brackets are % inhibition. Blanks mean that the value was not determined.

[0409] [Table D-2]

[0410] In pERK, + is IC 50 >0.3 μM, and ++ indicates IC of 0.03–0.3 μM. 50 indicates the range of IC 50 <0.03 μM. For proliferation, * indicates EC 50 indicates proliferation >3 μM, ** indicates EC 50 indicates the range, and *** indicates EC 50 Indicates <0.3 μM. N / A: Not available. Belv: Belvarafenib. Values ​​in brackets are % inhibition. Blank means value was not determined.

[0411] In the case of RAS mutant cancer cell lines, pERK IC 50 Curve Y 最小All % values ​​were above -20% and were considered to indicate minimal or no induction, therefore the compound does not cause detectable paradoxical activation of the pathway in this panel of cancer cell lines. In contrast, comparative results for berbalafenib (obtained from MedChem Express, catalog number HY-109080; CAS number 1446113-23-0) show mild to strong pathway induction in the same cell lines (Y in 10 of the 13 RAS mutant cell lines tested). 最小 <-30%).

[0412] (d) CellTiter-Glo (登録商標) Measurement of the growth inhibitory effect of the reagent on cultured human cancer cell lines (CCL)

[0413] CellTiter-Glo (登録商標) Viability analysis was performed on cells seeded in 96-well flat-bottom white opaque plates (Greiner or Croning) in 100 μL of complete RPMI-1640 growth medium at the densities indicated in Table E below (for each CCL, CellTiter-Glo (登録商標) Cell density (cells / cm) (number of cells seeded per well of a 96-well plate to perform cell viability assays) 2 ) is the number of cells per well of a 96-well plate (0.32 cm 2 ) Cells were maintained overnight at 37°C under 5% CO2 before being treated with serial dilutions of compounds for 3 days.

[0414] [Table E]

[0415] In the dilution series, 100 μL / well of test substance dilutions prepared in complete RPMI-1640 growth medium are added to the cells initially seeded in 100 μL of growth medium. The dilution series is selected to cover the range of 10 μM to 0.33 nM with 10 concentrations. If necessary (such as in the case of A375 cells, which are more sensitive to compounds), the initial concentration of 10 μM is reduced to 1 μM, and further dilutions are made accordingly. The final concentration of DMSO in the assay is set to 0.5%.

[0416] After 3 days of incubation, the growth medium was removed by aspiration, and 60 μL of diluted CellTiter-Glo (登録商標) Reagent (10 μL of CellTiter-Glo (登録商標) Reagent + 50 μL of PBS) was added to each well. Cells were lysed and incubated on a plate shaker for 5 minutes. (登録商標) After equilibration in the reagent and subsequent incubation at room temperature for 10 minutes, the luminescence signal was then acquired on a Synergy Neo2 plate reader (Biotek).

[0417] The effect of each compound dilution on the proliferation of cancer cell lines was expressed as % inhibition and calculated using the formula shown below: An internal 100% inhibition control (1 mM trametinib; Catalog No. HY-10999; MedChem Express; CAS No. 871700-17-3) was included in each plate and analyzed using CellTiter-Glo (登録商標) This was used as a measure of the signal background. The value obtained for trametinib was subtracted from each data point. The mean value of the DMSO (vehicle) control (set as 0% inhibition) was set and used to calculate the % inhibition.

number

[0418] The growth inhibitory potency of each compound was determined as an EC200 value by plotting the effect value for each data point in the dilution series using GraphPad Prism (V7.0) or the Dotmatics Screening Ultra platform, and fitting the resulting curve with a log(agonist) vs. response-variable slope (four parameter) function. 50 expressed as a value.

[0419] As shown in Tables D-1 and D-2, the active agents exhibit antiproliferative activity in a variety of NRAS-, KRAS-, and NF1-mutant cancer cell lines (i.e., SK-MEL 30, IPC298, HepG2, HCT-116, Lovo, SW620, SW480, NCI-H358, NCI-H2122, Calu-6, and Mewo; see Tables D-1 and D-2, and for genotypes, see Table B) representing a wide variety of tissue types. Antiproliferative activity is often even stronger in cell lines harboring BRAF driver mutations (A375, A101D, A2058, RKO, HT29, NCIH2087, NCIH1755, and NCIH1666) (Tables D-1 and D-2). Notably, the IC of pERK reduction in KRAS-mutant and BRAF-mutant cell lines was 50 Values ​​and EC of the antiproliferative activity of the substances 50 The values ​​correlate well with each other (Table D). Thus, the compounds of the present invention are effective against several tumor types and may be used for these and other indications. This demonstrates that the compounds described herein are useful in treating various types of tumors.

[0420] (e) Results

[0421] Tables 3 and 4 below summarize the structures and biological results of exemplary compounds, each of which is followed by its respective table summarizing the chemical characterization and synthesis methods of the compounds.

[0422] [Table 3] JPEG2025535134000104.jpg238170JPEG2025535134000105.jpg236170JPEG2025535134000106.jpg236170JPEG2025535134 000107.jpg237170JPEG2025535134000108.jpg241170JPEG2025535134000109.jpg242170JPEG2025535134000110.jpg37170

[0423] For pERK assays, + indicates IC of 1–10 μM. 50 ++ indicates IC range of 0.5-1 μM 50 indicates the range of IC 50 <0.5 μM. 最小 % values ​​are for each IC 50 Indicates the lowest point on the curve. 最小 IC values ​​above -20% 50 Compounds showing a curve show minimal or no induction and are considered not to cause detectable paradoxical activation of the pathway. For the BRAF biochemical kinase assay, * denotes IC 50 indicates IC >10 nM, ** indicates IC between 1 and 10 nM 50 indicates the range of IC 50 <1 nM. For the CRAF biochemical kinase assay, § indicates IC 50 >50 nM, and §§ indicates IC of 10–50 nM. 50 indicates the range of IC 50 <10 nM.

[0424] [Table A3-1] JPEG2025535134000112.jpg208170JPEG2025535134000113.jpg215170JPEG2025535 134000114.jpg195170JPEG2025535134000115.jpg227170JPEG2025535134000116.j pg224170JPEG2025535134000117.jpg203170JPEG2025535134000118.jpg203170JPE G2025535134000119.jpg227170JPEG2025535134000120.jpg207170JPEG20255351340 00121.jpg207170JPEG2025535134000122.jpg203170JPEG2025535134000123.jpg22 5170JPEG2025535134000124.jpg220170JPEG2025535134000125.jpg213170JPEG202 5535134000126.jpg219170JPEG2025535134000127.jpg228170JPEG20255351340001 28.jpg208170JPEG2025535134000129.jpg211170JPEG2025535134000130.jpg123170

[0425] [Table 4]

[0426] For pERK assays, + indicates IC of 1–10 μM. 50 ++ indicates IC of 0.5-1 μM 50 indicates the range of IC 50 <0.5 μM. 最小 % values ​​are for each IC 50 Indicates the lowest point on the curve. 最小 IC values ​​above -20% 50 Compounds showing a curve show minimal or no induction and are considered not to cause detectable paradoxical activation of the pathway. For the BRAF biochemical kinase assay, * denotes IC 50indicates IC >10 nM, ** indicates IC between 1 and 10 nM 50 indicates the range of IC 50 <1 nM. For the CRAF biochemical kinase assay, § indicates IC 50 >50 nM, and §§ indicates IC of 10–50 nM. 50 indicates the range of IC 50 <10 nM.

[0427] [Table 4-1]

[0428] Numerous modifications can be made to any of the above-described embodiments without departing from the scope of the present invention. All literature, patent or scientific literature mentioned herein is incorporated by reference in its entirety for all purposes.

Claims

1. A compound of formula I below, or a pharmaceutically acceptable salt or solvate thereof 【Chemical 1】 where: R 1 is substituted or not substituted, OR 3 , S.R. 3 , N.H. 2 , NHR 3 , N(R 3 ) 2 , C 3～8 Cycloalkyl, C 4～8 Heterocycloalkyl, C 6～10 Aryl and C 5～10 heteroaryl; R 2 is replaced by C 6 Aryl or C 5～10 Heteroaryl, substituted or unsubstituted C 4～8 Heterocycloalkyl, and N(R 3 ) 2 Selected from: R 3 is independently in each occurrence substituted or unsubstituted, C 1～8 Alkyl, C 3～8 Cycloalkyl, C 4～8 Heterocycloalkyl, C 6～10 Aryl and C 5～10 heteroaryl; X 1 is a halogen atom or an electron-withdrawing group; X 2 is selected from H, halogen atoms and electron-withdrawing groups; X 3 and X 4 are H, halogen atoms, electron-withdrawing groups, and C 1～3 Alkyl, C 3～4 Cycloalkyl and OC 1～3 alkyl.

2. R 2 is replaced by C 6 Aryl or C 5～10 The compound of claim 1 which is heteroaryl.

3. R 2 F, Cl, Br, CN, NO 2 , and substituted or unsubstituted, C 1～3 Alkyl, C 3～4 Cycloalkyl or OC 1～3 C substituted with at least one group selected from alkyl 6 The compound of claim 2 which is aryl.

4. R 2 is a group of the formula: 【Chemistry 2】 where: R 4 is H, F, Cl, Br, CN, and substituted or unsubstituted, C 1～3 Alkyl, C 3～4 Cycloalkyl or OC 1～3 alkyl; R 5 is H, F, Cl, CN, and substituted or unsubstituted, C 1～3 Alkyl, C 3～4 Cycloalkyl or OC 1～3 alkyl; R 6 H, F, Cl, Br, NO 2 , N.H. 2 , and substituted or unsubstituted, C 1～3 Alkyl, C 3～4 Cycloalkyl or OC 1～3 alkyl; R 7 is H, F, Cl, and substituted or unsubstituted C 1～3 alkyl; R 8 H, F, and substituted or unsubstituted C 1～3 alkyl; Or R 4 and R 5 or R 5 and R 6 together with their adjacent carbon atoms form a substituted or unsubstituted carbocyclic or heterocyclic ring, provided that the heterocyclic ring is not a benzoxazolinone; and (---) is R 2 represents the bond that serves as the point of attachment between and the rest of the molecule; where R 4 is H or F, then R 5 , R 6 , R 7 or R 8 is other than H or F; and where R 5 If is CN, then R 4 , R 6 , R 7 or R 8 At least one of is other than H.

5. R 4 But H, F, Cl, Br, Me, Et, CN, CHF 2 and CF 3 5. The compound of claim 4, selected from:

6. R 5 But, H, F, Me, CF 3 6. The compound according to claim 4 or 5, wherein the compound is selected from: CN and Cl.

7. R 6 is H, F, Cl, Br, and substituted or unsubstituted C 1～3 Alkyl, C 3～4 Cycloalkyl or OC 1～3 The compound according to any one of claims 4 to 6, wherein the aryl group is selected from alkyl.

8. R 6 The compound according to any one of claims 4 to 7, wherein is selected from H, F, Cl, Me, Et and OMe.

9. R 7 The compound according to any one of claims 4 to 8, wherein is selected from H, Me, F and Cl.

10. R 8 The compound according to any one of claims 4 to 9, wherein is selected from H, Me and F.

11. R 4 is Cl, and substituted or unsubstituted C 1～3 alkyl; R 5 is H, F, Cl, and substituted or unsubstituted C 1～3 alkyl; R 6 H, F, Cl, substituted or unsubstituted C 1～3 Alkyl and substituted or unsubstituted OC 1～3 alkyl; and R 7 and R 8 are H, The compound of claim 4.

12. R 4 But Cl and CH 3 12. The compound of claim 11 selected from:

13. R 5 F, Cl and CH 3 13. The compound according to claim 11 or 12, selected from:

14. R 6 The compound according to any one of claims 11 to 13, wherein is H or F.

15. R 6 Cl, substituted or unsubstituted C 1～3 Alkyl, or substituted or unsubstituted OC 1～3 The compound according to any one of claims 11 to 13, which is alkyl.

16. R 6 But CH 3 or OCH 3 16. The compound of claim 15, wherein:

17. R 2 is a group of the formula 【Chemistry 3】 where: X 5 However, NH, NC 1～3 Alkyl, NC 3～4 cycloalkyl, O and S; R 9 , R 10 , R 11 are each independently H, F, Cl, CN, and substituted or unsubstituted, C 1～3 Alkyl, C 3～4 Cycloalkyl, C(O)OC 1～3 Alkyl or OC 1～3 alkyl, with the proviso that R 9 and R 11 one of is H and the other is different from H; and (---) is R 2 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

18. R 2 is a group of the formula 【Chemistry 4】 where: X 5 However, NH, NC 1～3 Alkyl, NC 3～4 cycloalkyl, O and S; R 9 is F, Cl, CN, and substituted or unsubstituted, C 1～3 Alkyl, C 3～4 Cycloalkyl, C(O)OC 1～3 Alkyl or OC 1～3 alkyl; R 10 and R 12 are each independently H, F, Cl, CN, and substituted or unsubstituted, C 1～3 Alkyl, C 3～4 Cycloalkyl, C(O)OC 1～3 Alkyl or OC 1～3 alkyl; and (---) is R 2 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

19. R 9 and R 10 are each independently selected from F, Cl, CN, and substituted or unsubstituted, C 1～3 Alkyl, C 3～4 Cycloalkyl, C(O)OC 1～3 Alkyl or OC 1～3 19. The compound of claim 17 or 18, wherein the alkyl is selected from the group consisting of aryl, aryl, arylsulfonyl ...

20. R 9 and R 10 each independently represents Cl, and substituted or unsubstituted C 1～3 20. The compound of claim 19, wherein the alkyl is selected from:

21. R 9 and R 10 The compound of claim 19, wherein each of

22. X 5 A compound according to any one of claims 17 to 21, wherein is O or S, preferably S.

23. R 2 is a group of the formula 【Chemistry 5】 where: X 9 , X 10 , X 11 , X 12 and X 13 are independently selected from N and C, where X 9 , X 10 , X 11 , X 12 and X 13 at least one and not more than two of are N; and R 19 , R 20 , R 21 , R 22 and R 23 H, F, Cl, Br, CN, NO 2 , N.H. 2 , and substituted or unsubstituted, C 1～3 Alkyl, C 3～4 Cycloalkyl or OC 1～3 alkyl or their bonded X 9 , X 10 , X 11 , X 12 or X 13 does not exist when is N; where X 9 and X 13 At least one of is not N; where X 9 and X 13 when one of is N, the other is not N or CH; and (---) is R 2 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

24. R 2 is a group of the formula 【Chemistry 6】 where: R 13 represents independently in each occurrence F, Cl, and substituted or unsubstituted C 1～3 Alkyl, C 3～4 Cycloalkyl or C 1～3 alkoxy; n is an integer selected from 0 to 8; or n is 2 to 8, and two R 13 together with their adjacent carbon atoms, C 3～4 forming a cycloalkyl; and (---) is R 2 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

25. R 13 is F, Me, OMe, or CH 2 25. The compound of claim 24, wherein n is OMe and n is 1 or 2.

26. R 13 26. The compound of claim 24 or 25, wherein: is at the 3-position.

27. R 2 is N(R 3 ) 2 2. The compound of claim 1, wherein:

28. R 3 substituted or unsubstituted, C 1～8 Alkyl or C 3～8 28. The compound of claim 27, wherein the alkyl is selected from cycloalkyl.

29. R 2 The compound according to claim 1, wherein is selected from the groups B1 to B77 described in the specification.

30. R 2 The compound of claim 29, wherein is selected from groups B1 to B6 described in the specification.

31. R 1 But, OR 3 or SR 3 The compound according to any one of claims 1 to 30,

32. R 1 But, SR 3 32. The compound of claim 31 , wherein:

33. R 3 substituted or unsubstituted C 1～8 Alkyl (e.g., C 1～3 The compound according to any one of claims 1 to 32, wherein the aryl group is aryl, ...

34. R 1 substituted or unsubstituted C 5～6 The compound according to any one of claims 1 to 30, which is a heteroaryl group.

35. R 1 substituted or unsubstituted C 9 The compound according to any one of claims 1 to 30, which is a heteroaryl group.

36. R 1 is a substituted or unsubstituted group selected from imidazolyl, pyrazolyl, triazolyl, indolyl, indazolyl, benzimidazolyl, benzotriazolyl, pyrrolopyridinyl (e.g., pyrrolo[3,2-b]pyridinyl or pyrrolo[3,2-c]pyridinyl), pyrazolopyridinyl (e.g., pyrazolo[1,5-a]pyridinyl), purinyl, and imidazopyrazinyl (e.g., imidazo[4,5-b]pyrazinyl), preferably a substituted or unsubstituted group linked to the thiazolopyrimidine core through a nitrogen atom.

37. R 1 substituted or unsubstituted C 4～6 The compound according to any one of claims 1 to 30, which is a heterocycloalkyl group.

38. R 1 The compound according to any one of claims 1 to 30, wherein is a substituted or unsubstituted group selected from the following: 【Chemistry 7】 where (---) is R 1 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

39. R 1 39. The compound of claim 38, wherein: is a substituted or unsubstituted group selected from: 【Chemistry 8】 where (---) is R 1 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

40. R 1 OH, halogen atoms, CN, NO 2 , C 1～6 Alkyl, C 2～6 Alkenyl, C 2～6 Alkynyl, OC 1～6 Alkyl, C 5～10 Heteroaryl, C 3～10 Cycloalkyl, C 4～10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 ) 2 , SO 2 R 15 , SO 2 N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , N(R 16 )C(O)N(R 14 ) 2 , N(R 16 )SO 2 N(R 14 ) 2 , N(R 14 ) 2 , P(O)(R 15 ) 2 , C.H. 2 C(O)R 15 , C.H. 2 C(O)N(R 14 ) 2 , C.H. 2 SO 2 R 15 , C.H. 2 SO 2 N(R 14 ) 2 , C.H. 2 N(R 16 )C(O)R 15 , C.H. 2 N(R 16 )SO 2 R 15 , C.H. 2 N(R 16 )C(O)N(R 14 ) 2 , C.H. 2 N(R 16 )SO 2 N(R 14 ) 2 , and C.H. 2 N(R 14 ) 2 and is substituted with at least one substituent selected from where: R 14 independently in each occurrence, H, C 1～6 Alkyl, C 2～6 Alkenyl, C 2～6 Alkynyl, C 3～10 Cycloalkyl, C 4～10 Heterocycloalkyl, C 6 Aryl, and C 5～10 heteroaryl, or two R 14 together with their adjacent nitrogen atoms, form C 4～10 forming a heterocycloalkyl group; R 15 independently in each occurrence, C 1～6 Alkyl, C 2～6 Alkenyl, C 2～6 Alkynyl, C 3～10 Cycloalkyl, C 6 Aryl, and C 5～10 heteroaryl; and R 16 independently in each occurrence, H, C 1～6 Alkyl, C 2～6 Alkenyl, C 2～6 Alkynyl, C 3～10 Cycloalkyl, C 6 Aryl, and C 5～10 heteroaryl; wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group may be further optionally substituted. A compound according to any one of claims 1 to 39.

41. R 1 A compound according to any one of claims 1 to 30, wherein is a group of the formula 【Chemistry 9】 where: R 17 H, OH, halogen atoms, CN, NO 2 , C 1～6 Alkyl, C 2～6 Alkenyl, C 2～6 Alkynyl, OC 1～6 Alkyl, C 5～10 Heteroaryl, C 3～10 Cycloalkyl, C 4～10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 ) 2 , SO 2 R 15 , SO 2 N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , N(R 16 )C(O)N(R 14 ) 2 , N(R 16 )SO 2 N(R 14 ) 2 , N(R 14 ) 2 , P(O)(R 15 ) 2 , C.H. 2 C(O)R 15 , C.H. 2 C(O)N(R 14 ) 2 , C.H. 2 SO 2 R 15 , C.H. 2 SO 2 N(R 14 ) 2 , C.H. 2 N(R 16 )C(O)R 15 , C.H. 2 N(R 16 )SO 2 R 15 , C.H. 2 N(R 16 )C(O)N(R 14 ) 2 , C.H. 2 N(R 16 )SO 2 N(R 14 ) 2 , and C.H. 2 N(R 14 ) 2 Selected from: R 27 However, H, OH, halogen atoms, CN, NO 2 , C 1～6 Alkyl, C 2～6 Alkenyl, C 2～6 Alkynyl, OC 1～6 Alkyl, C 5～10 Heteroaryl, C 3～10 Cycloalkyl, C 4～10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 ) 2 , SO 2 R 15 , SO 2 N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , N(R 16 )C(O)N(R 14 ) 2 , N(R 16 )SO 2 N(R 14 ) 2 , N(R 14 ) 2 , P(O)(R 15 ) 2 , C.H. 2 C(O)R 15 , C.H. 2 C(O)N(R 14 ) 2 , C.H. 2 SO 2 R 15 , C.H. 2 SO 2 N(R 14 ) 2 , C.H. 2 N(R 16 )C(O)R 15 , C.H. 2 N(R 16 )SO 2 R 15 , C.H. 2 N(R 16 )C(O)N(R 14 ) 2 , C.H. 2 N(R 16 )SO 2 N(R 14 ) 2 , and C.H. 2 N(R 14 ) 2 and preferably H, a halogen atom, optionally substituted, C 1～6 alkyl, or optionally substituted OC 1～6 alkyl; X 6 is N or CH; and X 7 is N, and R 18 is not present; or X 7 is C, and R 18 But C 1～6 Alkyl, C 2～6 Alkenyl, C 2～6 Alkynyl, OC 1～6 Alkyl, C 5～10 Heteroaryl, C 3～10 Cycloalkyl, C 4～10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 ) 2 , SO 2 R 15 , SO 2 N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , N(R 16 )C(O)N(R 14 ) 2 , N(R 16 )SO 2 N(R 14 ) 2 , N(R 14 ) 2 , P(O)(R 15 ) 2 , C.H. 2 C(O)R 15 , C.H. 2 C(O)N(R 14 ) 2 , C.H. 2 SO 2 R 15 , C.H. 2 SO 2 N(R 14 ) 2 , C.H. 2 N(R 16 )C(O)R 15 , C.H. 2 N(R 16 )SO 2 R 15 , C.H. 2 N(R 16 )C(O)N(R 14 ) 2 , C.H. 2 N(R 16 )SO 2 N(R 14 ) 2 , and C.H. 2 N(R 14 ) 2 Selected from: where R 14 , R 15 and R 16 is as defined in claim 40; wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, or heteroaryl may be further optionally substituted; and where (---) is R 1 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

42. X 6 42. The compound of claim 41, wherein is N.

43. X 6 42. The compound of claim 41, wherein is CH.

44. X 7 is N and R 17 H, halogen atoms, OH, CN, C 1～6 Alkyl, C 2～6 Alkenyl, C 2～6 Alkynyl, OC 1～6 Alkyl, C 5～10 Heteroaryl, C 3～10 Cycloalkyl, C 4～10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 ) 2 , SO 2 R 15 , SO 2 N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , N(R 16 )C(O)N(R 14 ) 2 , N(R 16 )SO 2 N(R 14 ) 2 , N(R 14 ) 2 , P(O)(R 15 ) 2 , C.H. 2 C(O)R 15 , C.H. 2 C(O)N(R 14 ) 2 , C.H. 2 SO 2 R 15 , C.H. 2 SO 2 N(R 14 ) 2 , C.H. 2 N(R 16 )C(O)R 15 , C.H. 2 N(R 16 )SO 2 R 15 , C.H. 2 N(R 16 )C(O)N(R 14 ) 2 , C.H. 2 N(R 16 )SO 2 N(R 14 ) 2 , and C.H. 2 N(R 14 ) 2 and R 18 is absent, wherein said alkyl, said alkenyl, said alkynyl, said cycloalkyl, said heterocycloalkyl, or said heteroaryl is optionally further substituted.

45. R 17 But C 1～6 Alkyl, C 5～10 Heteroaryl, C 4～10 Heterocycloalkyl, N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , C(O)N(R 14 ) 2 , and SO 2 N(R 14 ) 2 45. The compound of claim 44, wherein said alkyl, said alkenyl, said alkynyl, said cycloalkyl, said heterocycloalkyl, or said heteroaryl is selected from the group consisting of:

46. R 17 But, H, F, NH 2 and optionally substituted C 5～10 Heteroaryl or C 4～10 heterocycloalkyl, preferably optionally substituted; C 5～10 Heteroaryl or C 4～10 45. The compound of claim 44, wherein the compound is selected from heterocycloalkyl.

47. R 17 optionally substituted C 4～10 heterocycloalkyl, wherein the heterocycloalkyl is monocyclic or bicyclic and contains 1 to 3 heteroatoms, preferably wherein X 7 The compound according to any one of claims 41 to 46, wherein is N.

48. The heterocycloalkyl may be F, OH, oxo, CN, C 1～4 Alkyl and OC 1～4 alkyl, wherein said C 1～4 Alkyl is (e.g., F, OH, OC 1～3 48. The compound of claim 47, which may be further optionally substituted (such as with alkyl).

49. 49. The compound of claim 47 or 48, wherein the heterocycloalkyl is selected from piperidine, piperazine, thiomorpholine, and morpholine groups, or bicyclic structures (bridged or spiro) containing a piperidine, piperazine, thiomorpholine, or morpholine ring.

50. X 7 The compound according to any one of claims 41 to 43, wherein is C.

51. R 18 But C 1-6 Alkyl, C 5～10 Heteroaryl, C 3～10 Cycloalkyl, C 4～10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 ) 2 , SO 2 R 15 , SO 2 N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , N(R 16 )C(O)N(R 14 ) 2 , N(R 16 )SO 2 N(R 14 ) 2 , N(R 14 ) 2 , P(O)(R 15 ) 2 , C.H. 2 C(O)R 15 , C.H. 2 C(O)N(R 14 ) 2 , C.H. 2 SO 2 R 15 , C.H. 2 SO 2 N(R 14 ) 2 , C.H. 2 N(R 16 )C(O)R 15 , C.H. 2 N(R 16 )SO 2 R 15 , C.H. 2 N(R 16 )C(O)N(R 14 ) 2 , C.H. 2 N(R 16 )SO 2 N(R 14 ) 2 , and C.H. 2 N(R 14 ) 2 51. The compound of claim 50, wherein said alkyl, said alkenyl, said alkynyl, said cycloalkyl, said heterocycloalkyl, said aryl, or said heteroaryl is further optionally substituted.

52. R 18 But C(O)N(R 14 ) 2 , SO 2 R 15 , and SO 2 N(R 14 ) 2 52. The compound of claim 51 selected from:

53. R 17 H, OH, halogen atoms, C 1～6 Alkyl, N(R 14 ) 2 and optionally substituted C 5～10 53. The compound of any one of claims 50 to 52, selected from heteroaryl.

54. R 17 But, H, F, NH 2 and optionally substituted C 5～10 Heteroaryl, preferably H, F, or NH 2 54. The compound of claim 53, wherein the compound is selected from:

55. R 14 is independently in each occurrence H, optionally substituted C 1～6 Alkyl, optionally substituted C 3～10 Cycloalkyl, optionally substituted C 4～10 Heterocycloalkyl and optionally substituted C 5～6 heteroaryl, or two R 14 together with their adjacent nitrogen atoms, optionally substituted C 4～10 A compound according to any one of claims 40 to 54, which forms a heterocycloalkyl group.

56. Two R's 14 together with their adjacent nitrogen atoms, optionally substituted C 4～10 56. The compound of claim 55, wherein the compound forms a heterocycloalkyl group, wherein the heterocycloalkyl is monocyclic or bicyclic and contains 1 to 3 heteroatoms.

57. The heterocycloalkyl may be F, OH, oxo, CN, C 1～4 Alkyl and OC 1～4 alkyl, wherein said C 1～4 Alkyl is (e.g., F, OH, OC 1～3 57. The compound of claim 56, which may be further optionally substituted (such as with alkyl).

58. 58. The compound of any one of claims 55 to 57, wherein the heterocycloalkyl is selected from piperidine, piperazine, thiomorpholine, and morpholine groups, or bicyclic structures (bridged or spiro) containing a piperidine, piperazine, thiomorpholine, or morpholine ring.

59. R 1 The compound according to any one of claims 1 to 30, wherein is selected from the group of formula 【Chemistry 10】 where R 14 , R 17 and R 27 is as defined above, and (---) is R 1 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

60. R 1 60. The compound of claim 59, wherein: 【Chemistry 11】 where R 14 , R 17 and R 27 is as defined above, and (---) is R 1 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

61. R 1 A compound according to any one of claims 1 to 30, wherein is a group of the formula 【Chemistry 12】 where: X 15 , X 16 , X 17 and X 18 are independently O, N, S and CR 17 where R 17 is as defined above; where X 15 , X 16 , X 17 and X 18 not more than two of are O, N, or S; and where (---) is R 1 represents the bond that serves as the point of attachment between the group and the rest of the molecule.

62. R 1 A compound according to any one of claims 1 to 30, wherein is selected from the groups A1 to A550 described in the specification.

63. R 1 is selected from the groups A1 to A3, A8, A19, A20, A22, A23, A25, A28, A29, A32 to A39, A60, A63 to A66, A69, A72 to A78, A81 to A83, A86, A89, A93, A96, A100, A101, A104, A105, A109 to A111, A113, A115, A118, A121 to A123, A127 and A132 as described herein.

64. R 1 is selected from the groups A1 to A3, A8, A19, A22, A25, A28, A29, A32, A36, A37, A64, A67, A74, A77, A78, A82, A83, A89, A96, A109, A110 and A111 described herein.

65. X 1 is Cl, and X 2 The compound of any one of claims 1 to 64, wherein is F.

66. X 1 is F, and X 2 The compound of any one of claims 1 to 64, wherein is H.

67. X 1 and X 2 The compound according to any one of claims 1 to 64, wherein each of

68. X 3 and X 4 68. The compound of any one of claims 1 to 67, wherein each is H.

69. X 3 is F, and X 4 The compound of any one of claims 1 to 67, wherein is H.

70. 68. The compound of claim 67, wherein the compound is a compound of formula II: 【Chemistry 13】 where R 1 , R 4 , R 5 and R 6 are each independently as defined above, preferably R 4 is selected from Cl, Br and methyl; R 5 is selected from H, F, Cl, and methyl; and R 6 is selected from H, F, Cl, Me and OMe.

71. 71. The compound of claim 70, wherein the compound is a compound of formula IV: 【Chemistry 14】 where X 6 , X 7 , R 4 , R 5 , R 6 , R 17 , R 27 and R 18 are each independently as defined above.

72. 71. The compound of claim 70, wherein the compound is a compound of formula V: 【Chemistry 15】 where R 4 , R 5 , R 6 , X 15 , X 16 , X 17 and X 18 are each independently as defined above.

73. 68. The compound of claim 67, wherein the compound is a compound of formula III: 【Chemistry 16】 where R 1 , R 9 , R 10 , R 12 and X 5 are each independently as defined above.

74. 74. The compound of claim 73, wherein the compound is a compound of formula VI: 【Chemistry 17】 where R 9 , R 10 , R 12 , R 17 , R 18 , R 27 , X 5 , X 6 and X 7 are each independently as defined above.

75. 74. The compound of claim 73, wherein the compound is a compound of formula VII: 【Chemistry 18】 where R 9 , R 10 , R 12 , X 5 , X 15 , X 16 , X 17 and X 18 are each independently as defined above.

76. 2. The compound according to claim 1, or a salt thereof and / or a solvate thereof, wherein the compound is selected from Examples 1 to 159 as defined herein.

77. 77. The compound according to claim 76, or a salt thereof and / or a solvate thereof, wherein the compound is selected from Examples 2, 4, 6, 7, 14, 16, 18, 30, 31, 33-37, 40, 43-46, 49, 51-60, 81, 84-88, 90, 93-99, 102-105, 108, 111, 112, 116-119, 122, 126, 127, 130, 131, 135-137, 139, 141, 144, 147, 148, 149, 153 and 158 described herein.

78. 77. The compound according to claim 76, or a salt thereof and / or a solvate thereof, wherein the compound is selected from Examples 4, 6, 7, 14, 16, 18, 30, 33, 35, 36, 37, 40, 43-45, 49, 51, 56-58, 85, 88, 95, 98, 99, 103, 104, 105, 111, 112, 116, 122, 135 and 136 described herein.

79. A pharmaceutical composition comprising a compound according to any one of claims 1 to 78, together with a pharmaceutically acceptable carrier, diluent or excipient.

80. 80. A method of using a compound according to any one of claims 1 to 78 for the treatment of a disease or disorder selected from a proliferative disease or disorder, a developmental abnormality caused by dysregulation of the RAS-ERK signalling cascade (RASopathy), or an inflammatory disease or immune system disorder.

81. 81. The method of claim 80, wherein the disease or disorder is selected from neoplasias and developmental abnormalities.

82. 82. The use of claim 80 or 81, wherein the disease or disorder is associated with a RAF gene mutation (e.g., ARAF, BRAF, or CRAF).

83. The use of any one of claims 80 to 82, wherein the disease or disorder is associated with a RAS gene mutation (e.g., KRAS).

84. The method of any one of claims 80 to 83, wherein the disease or disorder is associated with a mutation or amplification of a receptor tyrosine kinase (e.g., EGFR, HER2), or a mutation or amplification in a downstream regulator of the receptor, RAS (e.g., gain of function of SOS1, loss of function of NF1).

85. The use according to any one of claims 80 to 84, wherein the disease or disorder is a neoplasm.

86. 86. The method of claim 85, wherein the neoplasm is selected from melanoma, thyroid cancer (e.g., papillary thyroid carcinoma), colorectal cancer, ovarian cancer, breast cancer, endometrial cancer, liver cancer, sarcoma, gastric cancer, pancreatic cancer, Barrett's adenocarcinoma, glioma (e.g., ependymoma), lung cancer (e.g., non-small cell lung carcinoma), head and neck cancer, acute lymphocytic leukemia, acute myeloid leukemia, non-Hodgkin's lymphoma, and hairy cell leukemia.

87. 86. The use of claim 85, wherein the neoplasm is selected from colon or colorectal cancer, lung cancer, pancreatic cancer, thyroid cancer, breast cancer and melanoma.

88. 88. The use of any one of claims 80 to 87, wherein said treatment comprises inhibiting the RAS-ERK signaling pathway without substantial induction of paradoxical pathways.

89. 80. A method for treating a disease or disorder selected from a proliferative disease or disorder, a developmental abnormality caused by dysregulation of the RAS-ERK signaling cascade (RASopathy), or an inflammatory disease or immune system disorder, said method comprising administering to a subject in need of said treatment a compound according to any one of claims 1 to 78.

90. 90. The method of claim 89, wherein the disease or disorder is selected from neoplasia and developmental abnormalities.

91. 91. The method of claim 89 or 90, wherein the disease or disorder is associated with a RAF gene mutation (e.g., ARAF, BRAF, or CRAF).

92. 92. The method of any one of claims 89 to 91, wherein the disease or disorder is associated with a RAS mutation (e.g., KRAS).

93. 93. The method of any one of claims 89 to 92, wherein the disease or disorder is associated with a mutation or amplification of a receptor tyrosine kinase (e.g., EGFR, HER2), or a mutation or amplification in a downstream regulator of the receptor RAS (e.g., gain of function of SOS1, loss of function of NF1).

94. 94. The method of any one of claims 89 to 93, wherein the disease or disorder is a neoplasm.

95. 95. The method of claim 94, wherein the neoplasm is selected from melanoma, thyroid cancer (e.g., papillary thyroid carcinoma), colorectal cancer, ovarian cancer, breast cancer, endometrial cancer, liver cancer, sarcoma, gastric cancer, pancreatic cancer, Barrett's adenocarcinoma, glioma (e.g., ependymoma), lung cancer (e.g., non-small cell lung cancer), head and neck cancer, acute lymphocytic leukemia, acute myeloid leukemia, non-Hodgkin's lymphoma, and hairy cell leukemia.

96. 95. The method of claim 94, wherein the neoplasm is selected from colon or colorectal cancer, lung cancer, pancreatic cancer, thyroid cancer, breast cancer, and melanoma.

97. 97. The method of any one of claims 89 to 96, wherein the method comprises inhibiting the RAS-ERK signaling pathway without substantial induction of paradoxical pathways.

98. A method for inhibiting the abnormal proliferation of a cell, the method comprising contacting said cell with a compound according to any one of claims 1 to 78.

99. 99. The method of claim 98, wherein the cell comprises a mutated RAF protein kinase (e.g., a mutated ARAF, BRAF, or CRAF).

100. 100. The method of claim 98 or 99, wherein the cells contain a mutated RAS gene (e.g., a mutated KRAS).

101. 101. The method of any one of claims 98 to 100, wherein the abnormal proliferation is associated with a mutation or amplification of a receptor tyrosine kinase (e.g., EGFR, HER2), or a mutation or amplification in a RAS downstream regulator of the receptor (e.g., SOS1 gain of function, NF1 loss of function).

102. 102. The method of any one of claims 98 to 101, wherein the cell is selected from a melanoma cell, a thyroid cancer cell (e.g., a papillary thyroid carcinoma cell), a colorectal cancer cell, an ovarian cancer cell, a breast cancer cell, an endometrial cancer cell, a liver cancer cell, a sarcoma cell, a gastric cancer cell, a pancreatic cancer cell, a Barrett's adenocarcinoma cell, a glioma cell (e.g., an ependymoma cell), a lung cancer cell (e.g., a non-small cell lung carcinoma cell), a head and neck cancer cell, an acute lymphocytic leukemia cell, an acute myeloid leukemia cell, a non-Hodgkin's lymphoma cell, and a hairy cell leukemia cell.

103. 103. The method of any one of claims 98 to 102, wherein the cells are selected from colon or colorectal cancer cells, lung cancer cells, pancreatic cancer cells, thyroid cancer cells, breast cancer cells and melanoma cells.

104. 104. The method of any one of claims 98 to 103, wherein the method inhibits the RAS-ERK signaling pathway without substantial induction of paradoxical pathways.

105. 105. The method of any one of claims 98 to 104, wherein said contacting is carried out in vivo.

106. 105. The method of any one of claims 98 to 104, wherein said contacting is carried out ex vivo.

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