Thiazolo [5, 4-d] pyrimidine compounds, compositions comprising same and uses thereof

Abstract

Compounds, compositions and their use in the treatment of proliferative diseases or disorders, e.g., associated with RAF gene mutations and / or RAS gene mutations. The disclosed compounds are compounds of formula (I), or pharmaceutically acceptable salts and / or solvates thereof, where R1, R2, X1, X2, X3 and X4 are as defined herein: # imgabs0 #

Description

Technical Field

[0001] The present invention generally relates to thiazolo[5,4-d]pyrimidine compounds, pharmaceutical compositions comprising the same, and their use in treating and preventing diseases characterized by dysregulation of the RAS-ERK pathway (eg, cancer, RAS pathway diseases). Background Art

[0002] The RAS-RAF-MEK-ERK (RAS: rat sarcoma; RAF: rapidly accelerated fibrosarcoma; MEK: mitogen-activated protein kinase; ERK: extracellular signal-regulated kinase) signaling pathway (hereinafter referred to as the RAS-ERK pathway) plays a key role in transmitting proliferation signals generated by growth factor receptors from the plasma membrane to the nucleus. In most cancers, this pathway is dysregulated by activation of receptor tyrosine kinases (RTKs) (e.g., ERBB1, ERBB2, FLT3, RET, KIT), activation or inactivation of RAS regulators (SOS1 and NF1), and constitutive activating mutations in RAS genes (H-, K-, and NRAS; 30% in total in cancer) or BRAF genes (8% in cancer). The incidence of KRAS mutations is particularly high in pancreatic cancer (>90%), colorectal cancer (50%), and lung cancer (30%). For its part, BRAF mutations are particularly frequent in malignant melanoma (70%), thyroid cancer (40%), and colorectal cancer (10%) (mutation frequencies based on COSMIC (Catalogue of Somatic Mutations in Cancer) C atalogue O f S omatic M utations I n C ancer); Wellcome Trust Sanger Institute) released v95, November 24, 2021).

[0003] RAS protein is a small GTPase, which transmits extracellular growth signals to intracellular effectors to control important processes such as cell differentiation, proliferation and survival (Nat.Rev.Cancer 2003,3,459).After RTK stimulation, physiological activation of RAS occurs on the plasma membrane, resulting in the GTP load of GTPase, thereby causing its activation.Activated RAS interacts and activates a series of effector molecules, among which RAF kinase is the most critical RAS interactor in cancer development (Nature Rev.Drug Discov.2014,13,828).Carcinogenic mutations of glycine 12, glycine 13 or glutamine 61 in RAS isoforms lead to abnormal and constitutive signaling in human cancer (Nat.Rev.Cancer 2003,3,459) (COSMIC releases 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) (Nat. Rev. Mol. Cell Biol. 2015, 16, 281) and an N-terminal regulatory region (NTR) containing a RAS binding domain (RBD). In unstimulated cells, RAF proteins are sequestered in the cytoplasm as monomers. Binding of GTP-bound activated RAS to RBD induces membrane anchoring of RAF kinase (Nat. Rev. Mol. Cell Biol. 2015, 16, 281). At the same time, 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, resulting in a series of substrates being phosphorylated by ERK, thereby triggering cell-specific responses (Nat. Rev. Mol. Cell Biol. 2020 Oct;21(10),607).

[0005] To date, activating mutations in RAF isoforms have been primarily confined to the BRAF gene, although rare variants have been observed in ARAF and CRAF, underscoring the functional importance of this isoform (COSMIC publication v95, November 24, 2021). The most common cancer mutation in BRAF is the substitution of valine to glutamic acid at position 600 (known as BRAF V600E), enhances BRAF activity by stabilizing its active form (Cell 2004, 116, 855). In addition to the V600E allele, a variety of mutations have also occurred at other residues (e.g., G466V, D594G, etc.), which lead to enhanced RAF signaling through a variety of mechanisms (Nat. Rev. Mol. Cell Biol. 2015, 16, 281). According to their dependence on RAS activity and RAF dimerization, they are divided into three main categories (category 1 to category 3) (Nature, August 10, 2017; 548 (7666), 234-238). The key role of wild-type BRAF and CRAF in mediating RAS-driven tumorigenesis by stimulating ERK signaling has been widely verified (Cancer Cell 2011, 19, 652; Cancer Discov. 2012, 2, 685; Nat. Commun. 2017, 8, 15262). Thus, tumor cells are dependent on elevated and sustained signaling of the RAS-ERK pathway through both RAS and RAF activation, providing strong support for the concept of targeting RAF family kinases in cancer.

[0006] To address existing medical needs, a broad range of ATP-competitive RAF inhibitors have been developed over the past decade (Nat. Rev. Cancer 2017, 17, 676). Efforts have primarily focused on the most common RAS-independent BRAF mutations (BRAF V600E ), which led to the development and FDA approval of sulfonamide derivatives such as vemurafenib and dabrafenib. Some of these RAF inhibitors have been used in patients with recurrent BRAF V600E Alleles of BRAF have shown impressive efficacy in metastatic melanoma and have been approved for the treatment of this patient population (N. Engl. J. Med. 2011, 364, 2507; Lancet 2012, 380, 358). V600E Clinical responses in BRAF-dependent melanomas result from 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 always occurs, primarily due to reactivation of the RAS-ERK pathway (partially through mechanisms that stimulate RAF dimerization). These include upregulation of RTK signaling, RAS mutations, and BRAF V600E Amplification or truncation (Sci. Signal. 2010, 3, ra84; Nature 2010, 468, 973; Nature 2011, 480, 387; Nature Commun. 2012, 3, 724).

[0007] Meanwhile, tumors that exhibit RAS activity—due to activating RAS mutations or elevated RTK signaling but are otherwise BRAF wild-type—show resistance to BRAF. V600E In contrast, RAF inhibitors were found to induce ERK signaling in the presence of elevated RAS activity, thereby enhancing tumor cell proliferation (Nature 2010, 464, 431). This counterintuitive phenomenon, known as the paradoxical effect, has also been observed in normal tissues that rely on physiological RAS activity and underlies some of the adverse effects of RAF inhibitors observed in melanoma patients, such as the development of new secondary tumors (e.g., squamous cell carcinomas and keratoacanthomas) (Nat. Rev. Cancer 2014, 14, 455). Therefore, BRAF V600E Inhibitors are ineffective and even contraindicated for RAS-driven cancers. The potential mechanism stems from the ability of the compound to promote dimerization of the RAF kinase domain in the presence of active RAS (Nature 2010, 464, 431). This event is not limited to BRAF, but also involves other RAF family members and is determined by the compound binding mode and affinity (Nat.Chem.Biol.2013, 9, 428).

[0008] Two strategies have recently been pursued to circumvent the limitations of first-generation RAF inhibitors in RAS-mutated cancers. The first strategy relies on the observation that paradoxical ERK activation is a dose-dependent phenomenon, whereby induction occurs at subsaturating inhibitor concentrations, but the pathway is inhibited at saturating concentrations when the compound occupies both protomers of the RAF dimer. Therefore, this first strategy focused on developing molecules with higher binding affinity for all RAF paralogs in order to saturate RAF proteins at lower drug concentrations, thereby reducing paradoxical pathway induction (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 a strong RAF dimer induction ability and thus paradoxically stimulated RAS-ERK signaling, although to a lower extent than previous generations of RAF inhibitors. Although such compounds show improved properties, it has recently been found that most of them do not affect ARAF isoforms, which leads to paradoxical pathway activation and primary resistance, as well as acquired resistance in vitro and clinical settings (Clin Cancer Res. 2021, 27, 2061; Nature 2021, 594, 418). The second strategy involves designing compounds that bias the conformation of the BRAF kinase domain in an inactive state and thus do not paradoxically induce ERK signaling. This gave rise to the “Paradox Breaker” (PB) molecule PLX8394, a derivative of PLX4032 / vemurafenib (Nature 2015, 526, 583). These molecules retain the ability to target BRAF. V600E The high efficacy should therefore prove to be useful for the treatment of BRAF V600E However, while PLX8394 did not induce ERK signaling in the RAS mutant cell lines tested, it remained ineffective and had no effect on RAS mutant tumors.

[0009] There remains a need for inhibitors that effectively and consistently block RAS-ERK signaling and cell proliferation in human tumor cells carrying a variety of RAS and RAF genotypes. Importantly, it would be highly desirable to develop such inhibitors that also do not have contradictory pathway induction in multiple RAS mutant tumor cell lines. Summary of the invention

[0010] According to one aspect, the present invention relates to a compound of formula I, or a pharmaceutically acceptable salt or solvate thereof:

[0011]

[0012] in:

[0013] R 1 Selected from substituted or unsubstituted OR 3 , SR 3 NH 2 、NHR 3 、N(R 3 ) 2 , C 3-8 Cycloalkyl, C 4-8 Heterocycloalkyl, C 6-10 Aryl and C 5-10 heteroaryl;

[0014] R 2 Selected from substituted C 6 Aryl or C 5-10 Heteroaryl, substituted or unsubstituted C 4-8 Heterocycloalkyl and N(R 3 ) 2 ;

[0015] R 3 is independently selected at each occurrence from substituted or unsubstituted C 1-8 Alkyl, C 3-8 Cycloalkyl, C 4-8 Heterocycloalkyl, C 6-10 Aryl and C 5-10 heteroaryl;

[0016] X 1 is a halogen or an electron-withdrawing group;

[0017] X 2 is selected from H, halogen and electron withdrawing groups;

[0018] X 3 and X 4 Each is selected from H, halogen, electron withdrawing group, C 1-3 Alkyl, C 3-4 Cycloalkyl and OC 1-3 alkyl.

[0019] The compounds of formula I are further defined according to any of the embodiments, alone or in combination, and the Examples described throughout this specification.

[0020] According to another aspect, the present invention relates to the use of a pharmaceutical composition as defined in any one of the preceding embodiments, said composition comprising a compound as defined herein together with a pharmaceutically acceptable carrier, diluent or excipient.

[0021] In another aspect, the invention relates to the use of a compound as defined herein for treating a disease or disorder selected from a proliferative disease or disorder, a developmental abnormality caused by dysregulation of the RAS-ERK signalling cascade (RAS pathway disease), or an inflammatory disease or immune system disorder.

[0022] The present invention further relates to a method for treating a disease or condition selected from the following: a proliferative disease or condition, a developmental abnormality caused by dysregulation of the RAS-ERK signaling cascade (RAS pathway disease), or an inflammatory disease or immune system condition, comprising administering a compound as defined herein to a subject in need thereof. Also contemplated are methods for inhibiting abnormal cell proliferation, comprising contacting a cell with a compound as defined herein.

[0023] In one embodiment of the above-mentioned uses and methods, the disease or condition is selected from tumors and dysplasia, such as diseases or conditions associated with RAF gene mutations (e.g., ARAF, BRAF, or CRAF), diseases or conditions associated with RAS gene mutations (e.g., KRAS), or diseases or conditions associated with both RAF gene mutations and RAS gene mutations. In one embodiment, the disease or condition is associated with a mutation or amplification of a receptor tyrosine kinase (e.g., EGFR, HER2) or a mutation of a regulator of RAS downstream of the receptor (e.g., SOS1 gain of function, NF1 loss of function).

[0024] For example, the disease or disorder is a tumor, such as 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 lymphoblastic leukemia, acute myeloid leukemia, non-Hodgkin's lymphoma, and hairy cell leukemia. For example, the tumor is selected from colon cancer or colorectal cancer, lung cancer, pancreatic cancer, thyroid cancer, breast cancer, and melanoma. For example, any of the uses and methods of the present invention include inhibiting the RAS-ERK signaling pathway without substantially inducing a paradoxical pathway.

[0025] Additional objects and features of the compounds, compositions, methods and uses of the present invention will become more apparent upon reading the non-limiting description of the following illustrative embodiments and examples, which should not be construed as limiting the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Paradoxical induction of pERK signaling was not induced in RAS mutant HCT116 cells (Y MIN>-20%) of compounds as described herein (Example 44 and Example 122) and strongly induced this pathway in the same cell line (Y MIN Representative IC values ​​of the compound (PLX4720; CAS No. 918505-84-7) with a yield of -600%) 50 Inhibitory dose-response curves. DETAILED DESCRIPTION

[0027] All technical and scientific terms and expressions used herein have the same definitions as those commonly understood by those skilled in the art to which the invention belongs. However, the definitions of some of the terms and expressions used are provided below. If the definitions of terms in publications, patents and patent applications incorporated herein by reference are contrary to the definitions set forth in this specification, the definitions in this specification are subject. The section headings used herein are only for organizational purposes and should not be construed as limiting the disclosed subject matter.

[0028] i. definition

[0029] The chemical structures described herein are drawn according to conventional standards. In addition, when a drawn atom (e.g., a carbon atom) appears to include an incomplete valence, the valence is assumed to be satisfied by one or more hydrogen atoms, even if these are not necessarily explicitly drawn. Hydrogen atoms should be inferred to be part of the compound.

[0030] The terms used herein are only used for the purpose of describing specific embodiments and are not intended to be limiting. It should be noted that the singular forms "a", "an" and "the" also include plural forms, unless the context clearly stipulates otherwise. Thus, for example, a composition containing "a compound" is also covered by a mixture of two or more compounds. It should also be noted that the term "or" is usually used in its meaning including "and / or", unless the context clearly indicates otherwise. In addition, with respect to the terms "including", "includes", "having", "has", "with" or variations thereof used in the detailed description and / or claims, such terms are intended to be included in a manner similar to the term "comprising".

[0031] The term "about" or "approximately" means within an acceptable error range for a particular value 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, according to the practice in the art, "about" can mean within 1 or more than 1 standard deviation. Alternatively, "about" can mean a range of up to 20%, preferably up to 10%, more preferably up to 5%, and still more preferably up to 1% of a given value. Alternatively, particularly for biological systems or processes, the term can mean within an order of magnitude of a value, preferably within 5 times, and more preferably within 2 times. When describing specific values ​​in the present application and claims, unless otherwise stated, it should be assumed that the term "about" means within an acceptable error range for the specific value.

[0032] As used herein, the terms "compound", "compound described herein", "compound of the present application", "thiazolo[5,4-d]pyrimidine compound", "thiazolopyrimidine compound" and equivalent expressions refer to the compounds described in the present application, such as those covered by Structural Formula I, optionally with reference to any applicable embodiment, and also include exemplary compounds, such as the compounds of Examples 1 to 159 and their pharmaceutically acceptable salts, solvates, esters and prodrugs (when applicable). When a zwitterionic form is possible, for practical purposes, the compound can be drawn as its neutral form, but the compound is understood to also include its zwitterionic form. The embodiments herein may also exclude one or more compounds. A compound can be identified by its chemical structure or its chemical name. If the chemical structure and chemical name conflict, the chemical structure shall prevail.

[0033] Unless otherwise stated, structures depicted herein are also meant to include all isomeric (e.g., enantiomers, diastereomers, and geometric (or conformational)) forms of the structure, where applicable; for example, 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 invention are within the scope of the present specification. Unless otherwise stated, therapeutic compounds also encompass all possible tautomeric forms of the compounds shown, if any. The term also includes isotopically labeled compounds in which the atomic mass of one or more atoms is different from the most abundant atomic mass in nature. Examples of isotopes that can be incorporated into the compounds of the invention include, but are not limited to 2 H(D), 3 H(T), 11 C. 13 C. 14 C. 15 N. 18 O. 17O, any isotope of sulfur, etc. The compound can also exist in unsolvated form as well as solvated form, including hydrated form. The compound can exist in a variety of crystalline or amorphous forms. In general, all physical forms are equivalent for the intended uses herein and are intended to fall within the scope of the present invention.

[0034] When a particular enantiomer is preferred, in some embodiments, it may be substantially free of the corresponding enantiomer and may also be enantiomerically enriched. "Enantiomerically enriched" means that the compound is composed of a significantly larger proportion of one enantiomer. In certain embodiments, the compound is composed of at least about 90% by weight of the preferred enantiomer. In other embodiments, the compound is composed of at least about 95%, 98%, or 99% by weight of the preferred enantiomer. The preferred enantiomer can be separated from the racemic mixture by any method known to those skilled in the art, including high pressure liquid chromatography (HPLC) on a chiral support and the formation and crystallization of a chiral salt, or prepared by asymmetric synthesis.

[0035] The expression "pharmaceutically acceptable salts" refers to those salts of the compounds of the present invention which are suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic reactions, etc., within the scope of reasonable medical judgment, and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge, et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977). The salts can be prepared in situ during the final separation and purification of the compounds of the present invention, or prepared separately by reacting the free base functional group of the compound with a suitable organic or inorganic acid (acid addition salts) or by reacting the acid functional group of the compound with a suitable organic or inorganic base (base addition salts).

[0036] The term "solvate" refers to a physical association of one of the compounds of the present invention with one or more solvent molecules (including water and non-aqueous solvent molecules). This physical association may include hydrogen bonding. In some cases, for example, when one or more solvent molecules are incorporated into the lattice of a crystalline solid, the solvate will be able to separate. The term "solvate" encompasses both solution phase solvates and separable solvates. Exemplary solvates include, but are not limited to, hydrates, hemihydrates, ethanolates, hemiethanolates, n-propanolates, isopropanolates, 1-butanolates, 2-butanolates, and other physiologically acceptable solvents, such as International Conference on Harmonization (ICH), Guide for Industry, Q3CImpurities: Residual Solvents (1997) described in 3 types of solvents. Therefore, the compounds described herein also include each of their solvates and mixtures thereof.

[0037] As used herein, the expression "pharmaceutically acceptable ester" refers to an ester of a compound formed by the method of this specification sheet, which can be hydrolyzed in vivo, and includes those esters that are easily decomposed in the human body to leave the parent compound or its salt. Suitable ester groups include, for example, those derived from pharmaceutically acceptable aliphatic carboxylic acids, particularly alkanoic acids, alkenoic acids, cycloalkanoic acids and alkanedioic acids, wherein each alkyl or alkenyl moiety advantageously has no more than 6 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.

[0038] The expression "pharmaceutically acceptable prodrug" as used herein refers to those prodrugs of the compounds formed by the methods of the present invention, which are suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic reaction, etc., commensurate with a reasonable benefit / risk ratio, and effective for its intended use within the scope of reasonable medical judgment. "Prodrug" as used herein means a compound that can be converted in vivo by metabolic means (e.g., by hydrolysis) to provide any compound described by the formula of the present specification.

[0039] Abbreviations may also be used throughout the application, unless otherwise indicated, such abbreviations are intended to have the meaning generally understood in the art. The example of such abbreviations includes Me (methyl), Et (ethyl), Pr (propyl group), i-Pr (sec.-propyl), Bu (butyl), t-Bu (tert.-butyl), i-Bu (sec.-butyl), s-Bu (sec.-butyl), c-Bu (cyclobutyl), Ph (phenyl), Bn (benzyl), Bz (benzoyl), CBz or Cbz or Z (benzyloxycarbonyl), Boc or BOC (tert.-butoxycarbonyl) and Su or Suc (succinimide). In order to determine more, the additional definition of specific abbreviations is also included in the introduction of the embodiments section.

[0040] The number of carbon atoms in a hydrocarbyl substituent can be determined by the prefix "C x -C y " or "C x-y ” means, 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, when the prefix “C x -C y " or "C x-y When relating to a group that by definition incorporates one or more heteroatoms (e.g., heterocycloalkyl, heteroaryl, etc.), then x and y define, respectively, the minimum and maximum number of atoms in the ring, including carbon atoms and one or more heteroatoms.

[0041] As used herein, the term "alkyl" refers to a saturated, straight-chain or branched hydrocarbon group generally containing 1 to 20 carbon atoms. 1-8 The "alkyl" group contains 1 to 8 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl, heptyl, octyl groups and the like.

[0042] As used herein, the term "alkenyl" refers to a straight or branched chain hydrocarbon group containing one or more double bonds and typically containing 2 to 20 carbon atoms. 2-8 The term "alkenyl" refers to an alkenyl group containing 2 to 8 carbon atoms. Alkenyl groups include, but are not limited to, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, heptenyl, octenyl, and the like.

[0043] As used herein, the term "alkynyl" refers to a straight or branched chain hydrocarbon group containing one or more triple bonds and typically containing 2 to 20 carbon atoms. 2-8 "Alkynyl" contains 2 to 8 carbon atoms. Representative alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 1-butynyl, heptynyl, octynyl, and the like.

[0044] The terms "cycloalkyl", "alicyclic", "carbocycle", "carbocyclic" and equivalent expressions refer to groups containing saturated or partially unsaturated (non-aromatic) carbocyclic rings in monocyclic or polycyclic ring systems, including spirocyclic (sharing one atom), fused (sharing at least one bond) or bridged (sharing two or more bonds) carbocyclic ring systems, having three to fifteen 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]nonyl, norbornyl, and the like. The term cycloalkyl includes both unsubstituted cycloalkyl groups and substituted cycloalkyl groups. For example, the term "C 3-n "Cycloalkyl" refers to cycloalkyl groups having from 3 to the specified number "n" of carbon atoms in the ring structure. Unless the carbon number is otherwise specified, "lower cycloalkyl" as used herein has at least 3 and equal to or less than 8 carbon atoms in its ring structure.

[0045] As used herein, the terms "heterocycle", "heterocycloalkyl", "heterocyclyl", "heterocyclic group" and "heterocyclic ring" are used interchangeably and refer to a chemically stable 3- to 7-membered monocyclic or 7-10-membered bicyclic heterocyclic moiety that is saturated or partially unsaturated and has one or more, preferably 1 to 4, heteroatoms as defined above in addition to carbon atoms. The term "nitrogen" when used to refer to the ring atoms of the heterocycle includes substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 1 to 3 heteroatoms selected from oxygen, sulfur or nitrogen, the nitrogen can be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR (as in N-substituted pyrrolidinyl). The heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that produces a chemically stable structure, and any ring atom can be optionally substituted. Examples of heterocycloalkyl groups include, but are not limited to, 1,3-dioxolane, pyrrolidinyl, pyrrolidonyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydrodithienyl, tetrahydrothienyl, thiomorpholino, thioxanyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiol-cycloheptanyl, oxazepine Base, diazepine 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothiophenyl, dihydrofuranyl, 3-azabicyclo[3,1,0]hexanyl, 3-azabicyclo[4,1,0]heptanyl, quinolizinyl, quinuclidinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, etc. Heterocyclic groups also include groups in which the heterocyclic ring is fused to one or more aryl, heteroaryl, or alicyclic rings, such as indolinyl, 3H-indolyl, chromanyl, benzopyranyl, phenanthridinyl, 2-azabicyclo[2.2.1]heptanyl, octahydroindolyl, or tetrahydroquinolinyl, wherein the radical or point of attachment is on the heterocyclic ring. The heterocyclyl group may be monocyclic or bicyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions are independently optionally substituted. For example, the term "C 3-n "Heterocycloalkyl" refers to a heterocycloalkyl group having from 3 to the designated "n" number of atoms (including carbon atoms and heteroatoms) in the ring structure.

[0046] 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 is not aromatic. The term "partially unsaturated" is intended to encompass rings with multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties as defined herein.

[0047] The term "aryl", used alone or as part of a larger moiety (e.g., "aralkyl", "aralkyloxy", "aryloxy" or "aryloxyalkyl"), refers to an aromatic group having 4n+2 conjugated π (pi) electrons, wherein n is an integer from 1 to 3, in a monocyclic moiety or a bicyclic or tricyclic fused ring system having a total of 6 to 15 ring members, wherein at least one ring in the system is aromatic and wherein each ring in the system contains three to seven ring members. The term "aryl" can be used interchangeably with the term "aryl ring". In certain embodiments of the present specification, "aryl" refers to an aromatic ring system, which includes, but is not limited to, phenyl, biphenyl, naphthyl, azulenyl, anthracenyl, and the like, which may carry one or more substituents. The term "aralkyl" or "arylalkyl" refers to an alkyl residue attached to an aryl ring. Examples of aralkyl groups include, but are not limited to, benzyl, phenethyl, and the like. As used herein, the term "aryl" also includes within its scope groups in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, indenyl, phthalimidyl, naphthimidyl, fluorenyl, phenanthridinyl, or tetrahydronaphthyl, etc. For example, the term "C 6-n "Aryl" refers to an aromatic group having from 6 to the specified "n" number of atoms in the ring structure.

[0048] The term "heteroaryl", used alone or as part of a larger moiety (e.g., "heteroaralkyl" or "heteroaralkoxy"), refers to an aromatic group having 4n+2 conjugated π (pi) electrons, where n is an integer from 1 to 3 (e.g., having 5 to 18 ring atoms, preferably 5, 6 or 9 ring atoms; having 6, 10 or 14 π electrons shared in a cyclic array); and having 1 to 5 heteroatoms in addition to carbon atoms. The term "heteroatom" includes, but is not limited to, nitrogen, oxygen or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of basic nitrogen. A heteroaryl group can be a single ring or two or more fused rings. The term "heteroaryl" as used herein also includes groups in which a heteroaromatic ring is fused to one or more aryl, alicyclic or heterocyclic rings, wherein the attachment group or point of attachment is located on the heteroaromatic ring. Non-limiting examples of heteroaryl groups include thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridinyl, 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]pyridinyl or pyrrolo[3, [2,3-b]-1,4-oxazine-3(4H)-one. The heteroaryl group may be monocyclic or bicyclic. The heteroaryl group includes optionally substituted rings. The term "heteroaralkyl" refers to an alkyl group substituted by a heteroaryl group, wherein the alkyl and heteroaryl portions are independently optionally substituted. Examples include, but are not limited to, pyridylmethyl, pyrimidylethyl, and the like. For example, the term "C 5-n "Heteroaryl" refers to heteroaryl groups having from 5 to the specified number "n" of atoms (including carbon atoms and heteroatoms) in the ring structure.

[0049] As described herein, the compounds of the present invention may contain "optionally substituted" parts. In general, the term "substituted", whether or not it is preceded by the term "optionally", means that one or more hydrogens of the specified part are replaced by suitable substituents. Unless otherwise specified, the "optionally substituted" group can have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure can be substituted by more than one substituent selected from a specific group, the substituent can be the same or different at each position. The combination of substituents envisioned in this specification is preferably those that result in the formation of chemically stable or chemically feasible compounds. As used herein, the term "chemically stable" refers to a compound that does not substantially change when subjected to conditions that allow it to be produced, detected, and in some embodiments, recovered, purified, and used for one or more purposes disclosed herein.

[0050] The term "halogen" denotes a halogen atom, ie a fluorine, chlorine, bromine or iodine atom, preferably fluorine or chlorine.

[0051] The term "optionally substituted" refers to a group that is substituted or unsubstituted by replacing one, two, three or more hydrogen atoms thereon with substituents independently, including but not limited to F, Cl, Br, I, OH, CO 2 H, alkoxy, oxo, thioxo, NO 2 , CN, CF 3 NH 2 , NH alkyl, NH alkenyl, NH alkynyl, NH cycloalkyl, NH aryl, NH heteroaryl, NH heterocycle, dialkylamino, diarylamino, diheteroarylamino, O-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, CO 2 Alkyl, CO 2 Alkenyl, CO 2 Alkyne, CO 2 Cycloalkyl, CO 2 Aryl, CO 2 Heteroaryl, CO 2 Heterocycloalkyl, OC(O)alkyl, OC(O)alkenyl, OC(O)alkynyl, OC(O)cycloalkyl, OC(O)aryl, OC(O)heteroaryl, OC(O)heterocycloalkyl, C(O)NH 2 , C(O)NH alkyl, C(O)NH alkenyl, C(O)NH alkynyl, C(O)NH cycloalkyl, C(O)NH aryl, C(O)NH heteroaryl, C(O)NH heterocycloalkyl, OCO 2 Alkyl, OCO 2 Olefin, OCO2 Alkyne, OCO 2 Cycloalkyl, OCO 2 Aryl, OCO 2 Heteroaryl, OCO 2 Heterocycloalkyl, OC(O)NH 2 , OC(O)NH alkyl, OC(O)NH alkenyl, OC(O)NH alkynyl, OC(O)NH cycloalkyl, OC(O)NH aryl, OC(O)NH heteroaryl, OC(O)NH heterocycloalkyl, NHC(O) alkyl, NHC(O) alkenyl, NHC(O) alkynyl, NHC(O) cycloalkyl, NHC(O) aryl, NHC(O) heteroaryl, NHC(O) heterocycloalkyl, NHCO 2 Alkyl, NHCO 2 Alkenyl, NHCO 2 Alkynyl, NHCO 2 Cycloalkyl, NHCO 2 Aryl, NHCO 2 Heteroaryl, NHCO 2 Heterocycloalkyl, NHC(O)NH 2 , NHC(O)NH alkyl, NHC(O)NH alkenyl, NHC(O)NH alkenyl, NHC(O)NH cycloalkyl, NHC(O)NH aryl, NHC(O)NH heteroaryl, NHC(O)NH heterocycloalkyl, NHC(S)NH 2 , 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)NH 2 , NHC(NH)NH alkyl, NHC(NH)NH alkenyl, NHC(NH)NH alkenyl, NHC(NH)NH cycloalkyl, NHC(NH)NH aryl, NHC(NH)NH heteroaryl, NHC(NH)NH heterocycloalkyl, NHC(NH) alkyl, NHC(NH) alkenyl, NHC(NH) alkenyl, 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, C(NH)NH aryl, C(NH)NH heteroaryl, C(NH)NH heterocycloalkyl, 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)(O-alkenyl) 2 、P(O)(O-alkynyl) 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) 2 Alkyl, S(O) 2 Alkenyl, S(O) 2 Alkynyl, S(O) 2 Cycloalkyl, S(O) 2 Aryl, S(O)heteroaryl, S(O)heterocycloalkyl, SO 2 NH 2 、SO 2 NH alkyl, SO 2 NH alkenyl, SO 2 NH alkynyl, SO 2 NH cycloalkyl, SO 2 NH aryl, SO 2 NH heteroaryl, SO 2 NH heterocycloalkyl, NHSO 2 Alkyl, NHSO 2 Alkenyl, NHSO 2 Alkynyl, NHSO 2 Cycloalkyl, NHSO 2 Aryl, NHSO 2 Heteroaryl, NHSO 2 Heterocycloalkyl, CH 2 NH 2 , CH 2 SO 2 CH 3 , alkyl, alkenyl, alkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, cycloalkyl, carbocycle, heterocycle, polyalkoxyalkyl, polyalkoxy, methoxymethoxy, methoxyethoxy, SH, S-alkyl, S-alkenyl, S-alkynyl, S-cycloalkyl, S-aryl, S-heteroaryl, S-heterocycloalkyl, or methylthiomethyl.

[0052] ii.Compounds

[0053] The enumeration of a list of chemical groups in any definition of a variable herein includes the definition of the variable as any single group or combination of the listed groups. The narration of an embodiment of a variable herein includes the embodiment as any single embodiment or in combination with any other embodiment or portion thereof. The narration of an embodiment herein includes the embodiment as any single embodiment or in combination with any other embodiment or portion thereof. Therefore, if applicable, the following embodiments may exist alone or in combination.

[0054] The compounds of the present invention present 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 as defined herein are shown by general formula I:

[0055]

[0056] in:

[0057] R 1 Selected from substituted or unsubstituted OR 3 , SR 3 NH 2 、NHR 3 、N(R 3 ) 2 , C 3-8 Cycloalkyl, C 4-8 Heterocycloalkyl, C 6-10 Aryl and C 5-10 Heteroaryl, for example, selected from 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;

[0058] R 2 Selected from substituted C 6 Aryl or C 5-10 Heteroaryl, substituted or unsubstituted C 4-8 Heterocycloalkyl and N(R 3 ) 2 ;

[0059] R 3 is independently selected at each occurrence from substituted or unsubstituted C 1-8 Alkyl, C 3-8 Cycloalkyl, C 4-8 Heterocycloalkyl, C 6-10 Aryl and C 5-10 heteroaryl;

[0060] X 1 is a halogen or an electron-withdrawing group;

[0061] X 2 is selected from H, halogen and electron withdrawing groups;

[0062] X 3 and X 4 Each is selected from H, halogen, electron withdrawing group, C 1-3 Alkyl, C 3-4 Cycloalkyl and OC 1-3 alkyl;

[0063] or a pharmaceutically acceptable salt or solvate thereof.

[0064] For example, the electron withdrawing group is selected from a perhaloalkyl group (e.g., CF 3 or CCl 3 )、CN、NO 2 , sulfonates, alkylsulfonyls (such as SO 2 Me or SO 2 CF 3 ), alkylcarbonyl (e.g., C(O)Me), carboxylate, alkoxycarbonyl (e.g., C(O)OMe), and aminocarbonyl (e.g., C(O)NH 2 In one embodiment, X 1 is Cl and X 2 It is F or X 1 is F and X 2 is H, or X 1 and X 2 are all F. In another embodiment, X 3 and X 4 Each is H. In yet another embodiment, X 3 is F and X 4 It's H.

[0065] For example, the aminoarylsulfonamide moiety in formula I may be designated as L and is preferably selected from:

[0066]

[0067] Therein, the dashed line (---) represents the bond serving as the point of attachment between L and the rest of the molecule.

[0068] In yet another embodiment, R 2 is substituted C 6 Aryl or C 5-10 Heteroaryl, such as R 2 is C substituted with at least one group selected from 6 Aryl: F, Cl, Br, CN, NO2 , and substituted or unsubstituted C 1-3 Alkyl, C 3-4 Cycloalkyl or OC 1-3 Alkyl. For example, R 2 is a group of the formula:

[0069]

[0070] in:

[0071] R 4 is selected from H, F, Cl, Br, CN, and substituted or unsubstituted C 1-3 Alkyl, C 3-4 Cycloalkyl or OC 1-3 Alkyl groups, such as R 4 Selected from H, F, Cl, Br, Me, Et, CN, CHF 2 and CF 3 ;

[0072] R 5 is selected from H, F, Cl, CN, and substituted or unsubstituted C 1-3 Alkyl, C 3-4 Cycloalkyl or OC 1-3 Alkyl groups, such as R 5 Selected from H, F, Me, CF 3 , CN and Cl;

[0073] R 6 Selected from H, F, Cl, Br, NO 2 , NH 2 , and substituted or unsubstituted C 1-3 Alkyl, C 3-4 Cycloalkyl or OC 1-3 Alkyl groups, such as R 6 is selected from 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 the group consisting of H, F, Cl, Me, Et and OMe;

[0074] R 7 is selected from H, F, Cl, and substituted or unsubstituted C 1-3 Alkyl groups, such as R 7 is selected from the group consisting of H, Me, F and Cl;

[0075] R 8 is selected from H, F, and substituted or unsubstituted C 1-3 Alkyl groups, such as R 8 is selected from H, Me and F;

[0076] Or, R 4 and R 5 , or R 5 and R 6 Together with the adjacent carbon atoms, they form a substituted or unsubstituted carbocyclic or heterocyclic ring, provided that the heterocyclic ring (R 2 ) is not a benzoxazolinone; and

[0077] (---) indicates use as R 2 The bond between the point of attachment and the rest of the molecule;

[0078] Among them, when R 4 When it is H or F, R 5 , R 6 , R 7 or R 8 At least one of is not H or F; and

[0079] Among them, when R 5 When it is CN, then R 4 , R 6 , R 7 or R 8 At least one of them is not H.

[0080] 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 Each is H, and R 6 is selected from H, Cl, Me and OMe. In another embodiment, R 4 is selected from F, Cl and Me, R 6 , R 7 and R 8 Each is H, and R 5 Selected from F and Cl.

[0081] In another embodiment, R 4 is selected from Cl, and substituted or unsubstituted C 1-3 Alkyl (eg Me); R 5 is selected from H, F, Cl, and substituted or unsubstituted C 1-3 Alkyl (eg Me); R 6 is selected from H, and substituted or unsubstituted OC 1-3 Alkyl (e.g. OCH 3 ); and R 7 and R 8 Each is H.

[0082] In another embodiment, R4 is selected from H, Cl, Br and methyl; R 5 is selected from H, F and Cl; R 6 is selected from the group consisting of H, F, Cl, Me and OMe; and R 7 and R 8 Each is H.

[0083] In another embodiment, R 4 is selected from Cl, and substituted or unsubstituted C 1-3 Alkyl, preferably R 4 is Cl or Me; R 5 is selected from H, F, Cl, and substituted or unsubstituted C 1-3 Alkyl (such as Me), preferably R 5 is F, Cl or Me; R 6 is selected from H, F, Cl, substituted or unsubstituted C 1-3 Alkyl (such as Me), and substituted or unsubstituted OC 1-3 Alkyl (e.g. OCH 3 ), preferably R 6 It 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 Yes CH 3 , or OCH 3 ; and R 7 and R 8 Each is H. In yet another embodiment, R 6 is substituted C 1-3 alkyl.

[0084] In another example, R 2 is substituted C 5 Heteroaryl groups, for example groups of the formula:

[0085]

[0086] in:

[0087] X 5 Selected from NH, NC 1-3 Alkyl, NC 3-4 Cycloalkyl, O and S;

[0088] R 9 , R 10 , R 11 are each independently selected from H, F, Cl, CN, and substituted or unsubstituted C 1-3 Alkyl, C 3-4 Cycloalkyl, C(O)OC1-3 Alkyl or OC 1-3 Alkyl, provided that R 9 and R 11 One of is H and the other is not H; and

[0089] (---) indicates use as R 2 The bond at the point of attachment between the molecule and the rest of the molecule.

[0090] Alternatively, R 2 is a group of the formula:

[0091]

[0092] in:

[0093] X 5 Selected from NH, NC 1-3 Alkyl, NC 3-4 Cycloalkyl, O and S;

[0094] R 9 is 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;

[0095] R 10 and R 12 are each independently selected from 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

[0096] (---) indicates use as R 2 The bond at the point of attachment between the molecule and the rest of the molecule.

[0097] 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 In another embodiment, X 5 It is O or S, preferably S.

[0098] In another embodiment, R 2 is substituted C 5-10 Heteroaryl groups, for example groups of the formula:

[0099]

[0100] in:

[0101] X 9 , X 10 , X 11 , X 12 and X 13 are independently selected from N and C, wherein X 9 , X 10 , X 11 , X 12 and X 13 At least one and at most two of are N; and

[0102] R 19 , R 20 , R 21 , R 22 and R 23 Selected from H, F, Cl, Br, CN, NO 2 , NH 2 , and substituted or unsubstituted C 1-3 Alkyl, C 3-4 Cycloalkyl or OC 1-3 Alkyl groups, or when they are attached to X 9 , X 10 , X 11 , X 12 or X 13 When N is 19 , R 20 , R 21 , R 22 and R 23 does not exist;

[0103] Among them, X 9 and X 13 At least one of is not N;

[0104] Among them, X 9 and X 13 When one of is N, the other is not N or CH; and

[0105] (---) indicates use as R 2 The bond at the point of attachment between the molecule and the rest of the molecule.

[0106] In another example, R 2 It is C 5 Heterocycloalkyl. For example, R 2 is a group of the formula:

[0107]

[0108] in:

[0109] R 13 is independently selected at each occurrence from F, Cl, and substituted or unsubstituted C 1-3 Alkyl, C 3-4 Cycloalkyl or C 1-3 Alkoxy;

[0110] n is an integer selected from 0 to 8; or

[0111] n is 2 to 8, and both R 13 Together with their adjacent carbon atoms, they form C 3-4 cycloalkyl; and

[0112] (---) indicates use as R 2 The bond at the point of attachment between the molecule and the rest of the molecule.

[0113] In one embodiment, R 13 In another embodiment, R 13 Selected from F, Me, OMe and CH 2 OMe, and n is 1 or 2. For example, R 13 is a methoxy group at the 3-position, and n is 1.

[0114] In another example, R 2 is N(R 3 ) 2 For example, R 2 is N(R 3 ) 2 , and R 3 is selected from substituted or unsubstituted C 1-8 Alkyl or C 3-8 Cycloalkyl.

[0115] In another embodiment, the compound of Formula I is a compound of Formula II, or a pharmaceutically acceptable salt or solvate thereof:

[0116]

[0117] Among them, R 1 , R 4 , R 5 and R 6 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 6Selected from H, F, Cl, Me and OMe.

[0118] In yet another embodiment, the compound of Formula I is a compound of Formula III, or a pharmaceutically acceptable salt or solvate thereof:

[0119]

[0120] Among them, R 1 , R 9 , R 10 , R 12 and X 5 Each is independently as defined herein. 2 The groups are shown by Group B1 to Group B77 which are defined as follows:

[0121]

[0122]

[0123] (---) indicates use as R 2 The bond at the point of attachment between the molecule and the rest of the molecule.

[0124] In one embodiment, R 2 is selected from the groups B1 to B77, or preferably R 2 is selected from the groups B1 to B6.

[0125] In one embodiment of the compound of Formula I, R 1 Yes OR 3 or SR 3 , for example, R 1 Yes SR 3 In various embodiments, R 3 is substituted or unsubstituted C 1-8 Alkyl (e.g. C 1-3 alkyl).

[0126] In another embodiment, R 1 is substituted or unsubstituted C 6 In another embodiment, R 1 is substituted or unsubstituted C 4-6 Heterocycloalkyl. For example, R 1 is optionally substituted by one or two selected from halogen, OH, C 1-6 Alkyl and OC 1-6 Alkyl-substituted C 4-5 Heterocycloalkyl. For example, R 1 is N-pyrrolidinyl substituted by one or two groups selected from F and OH.

[0127] In another embodiment, R1 is substituted or unsubstituted C 5-6 Heteroaryl group, or substituted or unsubstituted C 9 In another embodiment, R 1 is a substituted or unsubstituted group selected from the group consisting of thienyl, imidazolyl, pyrazolyl, triazolyl, thiazolyl, pyridinyl, 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 the group consisting of 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 R 1 Attachment to the thiazolopyrimidine nucleus is via the nitrogen atom.

[0128] R 1 Examples of include substituted or unsubstituted groups selected from the following:

[0129]

[0130] Among them, (---) indicates that it is used as R 1 The bond at the point of attachment between the molecule and the rest of the molecule.

[0131] For example, R 1 is a substituted or unsubstituted group selected from the following:

[0132]

[0133] Among them, (---) indicates that it is used as R 1 The bond at the point of attachment between the molecule and the rest of the molecule.

[0134] In one embodiment, R 1 is one of the above groups further substituted by at least one substituent selected from the group consisting of OH, halogen, CN, NO 2 , C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, OC 1-6 Alkyl, C 5-10 Heteroaryl, C 3-10Cycloalkyl, 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 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N(R 14 ) 2 , CH 2 N(R 16 )C(O)R 15 , CH 2 N(R 16 )SO 2 R 15 , CH 2 N(R 16 )C(O)N(R 14 ) 2 , CH 2 N(R 16 )SO 2 N(R 14 ) 2 and CH 2 N(R 14 ) 2 ;

[0135] in:

[0136] R 14 independently selected at each occurrence from H, C1-6 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, they form C 4-10 Heterocycloalkyl groups;

[0137] R 15 In each occurrence, independently selected from 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

[0138] R 16 independently selected at each occurrence from 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;

[0139] Among them, including R 1 (including R 14 , R 15 and R 16 The alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group in the definition of ) is optionally further substituted.

[0140] In another embodiment, R 1 is a group of the formula:

[0141]

[0142] in:

[0143] R 17 Selected from H, OH, halogen, 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 、SO2 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 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N(R 14 ) 2 , CH 2 N(R 16 )C(O)R 15 , CH 2 N(R 16 )SO 2 R 15 , CH 2 N(R 16 )C(O)N(R 14 ) 2 , CH 2 N(R 16 )SO 2 N(R 14 ) 2 and CH 2 N(R 14 ) 2 ;

[0144] R 27 Selected from H, OH, halogen, CN, NO 2 , C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, OC 1-6 Alkyl, C 5-10 Heteroaryl, C 3-10Cycloalkyl, 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 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N(R 14 ) 2 , CH 2 N(R 16 )C(O)R 15 , CH 2 N(R 16 )SO 2 R 15 , CH 2 N(R 16 )C(O)N(R 14 ) 2 , CH 2 N(R 16 )SO 2 N(R 14 ) 2 and CH 2 N(R 14 ) 2 , preferably H, halogen (e.g., F), optionally substituted C 1-6 alkyl, or optionally substituted OC 1-6 alkyl;

[0145] X6 is N or CH; and

[0146] X 7 is N and R 18 does not exist; or

[0147] X 7 is C and R 18 Selected from 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 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N(R 14 ) 2 , CH 2 N(R 16 )C(O)R 15 , CH 2 N(R 16 )SO 2 R 15 , CH 2 N(R 16 )C(O)N(R14 ) 2 , CH 2 N(R 16 )SO 2 N(R 14 ) 2 and CH 2 N(R 14 ) 2 ;

[0148] Among them, R 14 , R 15 and R 16 As defined above;

[0149] Among them, including R 1 (including R 14 , R 15 , R 16 , R 17 and R 18 The alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl or heteroaryl groups in the definition of ) are optionally further substituted; and

[0150] Among them, (---) indicates that it is used as R 1 The bond at the point of attachment between the molecule and the rest of the molecule.

[0151] In another embodiment, R 1 is a group of the formula:

[0152]

[0153] in:

[0154] X 15 , X 16 , X 17 and X 18 Independently selected from O, N, S and CR 17 , where R 17 As defined above;

[0155] Among them, X 15 , X 16 , X 17 and X 18 At most two of are O, N, or S; and

[0156] Among them, (---) indicates that it is used as R 1 The bond at the point of attachment between the molecule and the rest of the molecule.

[0157] In one embodiment, the compound of Formula I is a compound of Formula IV or Formula V, or a pharmaceutically acceptable salt or solvate thereof:

[0158]

[0159] Among them, 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 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 Selected from H, Cl, F, Me and OMe.

[0160] In another embodiment, the compound of Formula I is a compound of Formula VI or Formula VII, or a pharmaceutically acceptable salt or solvate thereof:

[0161]

[0162] Among them, R 9 , R 10 , R 12 , R 17 , R 18 , R 27 , X 5 , X 6 , X 7 , X 15 , X 16 , X 17 and X 18 Each is independently as defined herein.

[0163] In one embodiment of the above formula, X 6 is N. In another embodiment, X 6 It is CH.

[0164] In another embodiment, X 7 Yes N, R 17 Selected from H, OH, halogen, 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 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N(R 14 ) 2 , CH 2 N(R 16 )C(O)R 15 , CH 2 N(R 16 )SO 2 R 15 , CH 2 N(R 16 )C(O)N(R 14 ) 2 , CH 2 N(R 16 )SO 2 N(R 14 ) 2 and CH 2 N(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 is optionally further substituted, preferably R 17 Selected from C 1-6 Alkyl, C 5-10Heteroaryl, 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 , where R 14 , R 15 , R 16 or R 17 The alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl or heteroaryl in is optionally further substituted. 17 Selected from H, F, NH 2 , 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 Heterocycloalkyl.

[0165] In another embodiment, R 17 is an optionally substituted C 4-10 Heterocycloalkyl, wherein the heterocycloalkyl may be monocyclic or bicyclic and include 1 to 3 heteroatoms, preferably wherein X 7 is N. In a preferred embodiment, the heterocycloalkyl group is, for example, replaced by at least one selected from F, OH, oxo, CN, C 1-4 Alkyl and OC 1-4 Alkyl group substituted, wherein the C 1-4 The alkyl group is optionally further substituted (e.g., by F, OH, OC 1-3 For example, the heterocycloalkyl group may be selected from optionally substituted piperidine, piperazine, thiomorpholine and morpholine groups, or a bicyclic structure (bridged or spiro) containing a piperidine, piperazine, thiomorpholine or morpholine ring.

[0166] In another embodiment, X 7 is C, for example X 7 is C and R 18 Selected from 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 、SO2 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 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N(R 14 ) 2 , CH 2 N(R 16 )C(O)R 15 , CH 2 N(R 16 )SO 2 R 15 , CH 2 N(R 16 )C(O)N(R 14 ) 2 , CH 2 N(R 16 )SO 2 N(R 14 ) 2 and CH 2 N(R 14 ) 2 , where R 14 , R 15 , R 16 or R 18 The alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, or heteroaryl in is optionally further substituted, preferably R 18 Selected from C(O)N(R 14 ) 2 、SO 2 R15 and SO 2 N(R 14 ) 2 In subclasses of these embodiments, R 17 Selected from H, halogen, OH, C 1-6 Alkyl, N(R 14 ) 2 , and optionally substituted C 5-10 Heteroaryl. For example, R 17 Selected from H, F, NH 2 , and optionally substituted C 5-10 Heteroaryl, preferably H, F or NH 2 .

[0167] In yet another embodiment, R 14 is independently selected at each occurrence from 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, they form C 4-10 Heterocycloalkyl group.

[0168] In another embodiment, R 17 is N(R 14 ) 2 , wherein the R 14 Together with their adjacent nitrogen atoms, they form C 4-10 A heterocycloalkyl group, wherein the heterocycloalkyl group may be monocyclic or bicyclic and include 1 to 3 heteroatoms, preferably wherein X 7 is N. In a preferred embodiment, the heterocycloalkyl group is, for example, replaced by at least one selected from F, OH, oxo, CN, C 1-4 Alkyl and OC 1-4 Alkyl group substituted, wherein the C 1-4 The alkyl group is optionally further substituted (e.g., by F, OH, OC 1-3 For example, the heterocycloalkyl group may be selected from optionally substituted piperidine, piperazine, thiomorpholine and morpholine groups, or a bicyclic structure (bridged or spiro) containing a piperidine, piperazine, thiomorpholine or morpholine ring.

[0169] In another embodiment, R 1 Selected from:

[0170]

[0171] Among them, R 14 , R 17 and R27 As defined herein, and (---) indicates use as R 1 The bond at the point of attachment between the molecule and the rest of the molecule.

[0172] In yet another embodiment, R 1 Selected from:

[0173]

[0174] Among them, R 14 , R 17 and R 27 As defined herein, and (---) indicates use as R 1 The bond at the point of attachment between the molecule and the rest of the molecule.

[0175] Additional sub-general embodiments are provided in the Examples section, wherein each substituent group R 1 (group), R 2 (B group), and L are as defined. Examples of combinations are also further described below and in Tables 3 and 4. Representative preferred compounds of Examples 1 to 159 are also described herein.

[0176] More specifically, exemplary R 1 The groups are shown as A1 to A550 as defined below:

[0177]

[0178]

[0179]

[0180]

[0181]

[0182]

[0183]

[0184]

[0185]

[0186]

[0187]

[0188]

[0189]

[0190]

[0191]

[0192]

[0193]

[0194]

[0195]

[0196]

[0197]

[0198] Among them, (---) indicates that it is used as R 1 The bond at the point of attachment between the molecule and the rest of the molecule.

[0199] In one embodiment, R 1 is selected from the group A1 to A550, or R 1 is selected from the group consisting of 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 group consisting of 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.

[0200] In one embodiment, R 1 is selected from the group consisting of 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 1is selected from the group consisting of 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.

[0201] The following embodiments describe R 1 (A1 to A550), R 2 Combinations of (B1 to B77) and L (L1 to L4) groups, which can be combined to produce compounds of Formula I.

[0202] A1-L-B1; A1-L-B2; A1-L-B3; A1-L-B4 to B75; A1-L-B76; A1-L-B77;

[0203] A2-L-B1; A2-L-B2; A2-L-B3; A2-L-B4 to B75; A2-L-B76; A2-L-B77;

[0204] A3-L-B1; A3-L-B2; A3-L-B3; A3-L-B4 to B75; A3-L-B76; A3-L-B77;

[0205] A4 to A548-L-B1; A4 to A548-L-B2; A4 to A548-L-B3; A4 to A548-L-B4 to B75; A4 to A548-L-B76; A4 to A548-L-B77;

[0206] A549-L-B1; A549-L-B2; A549-L-B3; A549-L-B4 to B75; A549-L-B76; A549-L-B77;

[0207] A550-L-B1; A550-L-B2; A550-L-B3; A550-L-B4 to B75; A550-L-B76; A550-L-B77;

[0208] Exemplary compounds as defined herein include each individual compound encompassed in Table 3 and Table 4 under Example 1 to Example 159.

[0209] Examples of preferred compounds are Examples 2, 4, 6, 7, 14, 16, 18, 30, 31, 33 to 37, 40, 43 to 46, 49, 51 to 60, 81, 84 to 88, 90, 93 to 99, 102 to 105, 108, 111, 112, 116 to 119, 122, 126, 127, 130, 131, 135 to 137, 139, 141, 144, 147, 148, 149, 153 and 158, or salts and / or solvates thereof.

[0210] Examples of more preferred compounds include Examples 4, 6, 7, 14, 16, 18, 30, 33, 35, 36, 37, 40, 43 to 45, 49, 51, 56 to 58, 85, 88, 95, 98, 99, 103, 104, 105, 111, 112, 116, 122, 135 and 136, or salts and / or solvates thereof.

[0211] It should be understood that any of the above compounds can be any amorphous, crystalline or polymorphic form, including any salt or solvate form, or a mixture thereof. The compound of this specification can be further modified by attaching various functional groups via any synthetic means described herein to enhance selective biological properties. Such modifications are known in the art, and include increasing the biological penetration of a given biological system (e.g., blood, lymphatic system, central nervous system), increasing oral availability, increasing solubility to allow administration by injection, changing metabolism and changing excretion rate.

[0212] These compounds can be prepared by conventional chemical synthesis, such as those illustrated in the schemes and embodiments of the present invention. As will be appreciated by those skilled in the art, other methods for synthesizing compounds of the chemical formula herein are obvious to those of ordinary skill in the art. In addition, various synthesis steps can be carried out in alternating order or sequence to obtain desired compounds.

[0213] iii. Methods, uses, formulations and administration

[0214] As used herein, the term "effective amount" means an amount of a drug or pharmaceutical agent that will elicit a biological or medical response to a tissue, system, animal, or human being that is being sought, for example, by a researcher or clinician. In addition, the term "therapeutically effective amount" means any amount that results in the treatment, cure, prevention, or improvement of a disease, disorder, or symptom thereof, or a decrease in the rate of progression of a disease or disorder, as compared to a corresponding subject that has not received such an amount. The term also includes within its scope amounts that are effective to enhance normal physiological function.

[0215] As used herein, the terms "treatment", "treat" and "treating" refer to reversing, alleviating, delaying the onset of, or inhibiting the progression of a disease or condition as described herein, or one or more symptoms thereof. In some embodiments, treatment may be administered after one or more symptoms have occurred. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual before the onset of symptoms (e.g., based on a history of symptoms and / or based on genetic or other predisposing factors). Treatment may also be continued after symptoms subside, for example to prevent or delay their recurrence.

[0216] In one embodiment, the disease or disorder 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 (RAS pathway disease), an inflammatory disease, or an immune system disorder.

[0217] According to some embodiments, the proliferative disease or condition to be treated is a tumor, an inflammatory disease or disorder, or a developmental abnormality involving constitutively activating mutations in the RAS and / or RAF genes (e.g., KRAS and / or ARAF, BRAF or CRAF mutations). The disease or condition may also be further associated with a mutation or amplification of a receptor tyrosine kinase (e.g., EGFR, HER2) or a mutation in a regulator of RAS downstream of the receptor (e.g., SOS1 gain of function, NF1 loss of function). For example, a compound as defined herein is an inhibitor of a signaling enzyme (e.g., B- and CRAF), which is not only effective in cells carrying RAF mutations (e.g., BRAF V600E The compounds of the invention are involved in controlling cell proliferation in tumors with RAs and, importantly, in the context of mutant RAS-driven cancers. Therefore, the compounds of the invention are useful, for example, in the treatment of diseases associated with the activity of these signaling enzymes and characterized by excessive or abnormal cell proliferation.

[0218] 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 involve cells with autonomous growth capacity, i.e., an abnormal state of disorder characterized by rapidly proliferating cell growth, which often forms distinct masses that exhibit partial or complete lack of structural organization and functional coordination with normal tissue.

[0219] For example, proliferative disorders or diseases are defined as "neoplasm," "neoplastic disorder," "neoplasm," "cancer," and "tumor," which terms are collectively intended to encompass hematopoietic neoplasms (e.g., lymphomas or leukemias) as well as solid tumors (e.g., sarcomas or carcinomas), including all types of precancerous and cancerous growths or oncogenic processes, metastatic tissues, or malignantly transformed cells, tissues, or organs, regardless of histopathological type or stage of invasion. Hematopoietic neoplasms are malignancies that affect hematopoietic structures (structures associated with blood cell formation) and components of the immune system, including leukemias (associated with leukocytes (white blood cells) and their precursors in the blood and bone marrow) derived from myeloid, lymphoid, or erythroid lineages, and lymphomas (associated with lymphocytes). Solid tumors include sarcomas, which are malignant tumors that originate in connective tissues such as muscle, cartilage, blood vessels, fibrous tissue, fat, or bone. Solid tumors also include cancers, i.e., malignant tumors produced by epithelial structures, including external epithelia (e.g., the endometrium of the skin and gastrointestinal tract, lungs, and cervix) and internal epithelia lining various glands (e.g., breast, pancreas, thyroid). Examples of tumors include leukemia and hepatocellular carcinoma, sarcoma, hemangioendothelial carcinoma, breast cancer, central nervous system cancer (e.g., astrocytoma, gliosarcoma, neuroblastoma, oligodendroglioma, and glioblastoma), prostate cancer, lung cancer and bronchial cancer, laryngeal cancer, esophageal cancer, colon cancer, colorectal cancer, gastrointestinal cancer, melanoma, ovarian cancer and endometrial cancer, renal cancer and bladder cancer, liver cancer, endocrine cancer (e.g., thyroid) and pancreatic cancer. For example, the disease or condition is selected from colon cancer, lung cancer, pancreatic cancer, thyroid cancer, breast cancer, and skin cancer. Examples of tumors include melanoma, papillary thyroid carcinoma, colorectal cancer, ovarian cancer, breast cancer, endometrial cancer, liver cancer, sarcoma, gastric cancer, Barrett's gland carcinoma, glioma (including ependymoma), lung cancer (including non-small cell lung cancer), head and neck cancer, acute lymphoblastic leukemia, acute myeloid leukemia, non-Hodgkin lymphoma, and hairy cell leukemia.

[0220] In an embodiment, the patient presenting one of the above-mentioned hematopoietic system or solid tumors has previously been treated with a RAS-ERK pathway targeted inhibitor (including an RTK, RAF, MEK or ERK inhibitor) but has developed resistance to the inhibitor. Inhibitors include standard of care treatments such as vemurafenib, dabrafenib, cobimetinib, trametinib, YERVOY, OPDIVO or any combination of these agents.

[0221] In an embodiment, the disease to be treated is defined as a developmental abnormality caused by dysregulation of the RAS-ERK signaling cascade (RAS pathway disorders: e.g., Noonan syndrome, Costello syndrome, LEOPARD syndrome, cardiofaciocutaneous syndrome, and hypertrophic cardiomyopathy).

[0222] In an embodiment, the disease to be treated is defined as an inflammatory disease or an immune system disorder. Examples of such inflammatory diseases or immune system disorders include inflammatory bowel disease, Crohn's disease, ulcerative colitis, systemic lupus erythematosus (SLE), rheumatoid arthritis, multiple sclerosis, thyroiditis, type 1 diabetes, sarcoidosis, psoriasis, allergic rhinitis, asthma, COPD (chronic obstructive pulmonary disease).

[0223] In one embodiment, the compounds as defined herein are inhibitors of RAS-ERK signaling and cell proliferation in tumor cells carrying at least one mutated RAS or RAF genotype, without inducing or substantially inducing paradoxical pathways.

[0224] As used herein, the term "patient or subject" refers to an animal, such as a mammal. Thus, a subject may refer to, for example, mice, rats, dogs, cats, horses, cows, pigs, guinea pigs, primates including humans, etc. Preferably, the subject is a human.

[0225] Therefore, the present specification also relates to a method of treating a subject (e.g., a human subject) suffering from a proliferative disease or disorder (e.g., a RAF-mutated and / or mutated RAS-driven cancer). The method comprises administering to a subject in need of such treatment a therapeutically effective amount of a compound as defined herein.

[0226] In certain embodiments, this specification provides a method for treating a disease (as described herein) of a subject, comprising administering a compound of this specification to a subject identified as being in need thereof. The identification of those patients who need to treat the above-mentioned disease is completely within the capabilities and knowledge of those skilled in the art. The medical field recognizes certain methods for identifying patients at risk of developing the above-mentioned disease that can be treated by the subject method, such as the family history of the subject patient and the risk factors associated with the development of the disease state. Medical personnel in this area can easily identify such candidate patients by using, for example, clinical tests, physical examinations, medical history / family history, and genetic determinations.

[0227] The method for evaluating the therapeutic efficacy of a subject includes determining the pre-treatment symptoms of the disease by methods well known in the art, and then administering a therapeutically effective amount of the compound of the present invention to the subject. After an appropriate period of time (e.g., 1 week, 2 weeks, 1 month, 6 months) after the administration of the compound, the symptoms of the disease are determined again. The regulation (e.g., reduction) of the symptoms of the disease and / or biomarkers (e.g., pERK or pMEK) indicates the efficacy of the treatment. The symptoms and / or biomarkers of the disease can be regularly determined throughout the treatment process. For example, the symptoms and / or biomarkers of the disease can be checked every few days, weeks, or months to evaluate the further efficacy of the treatment. The reduction of the symptoms of the disease and / or biomarkers indicates that the treatment is effective.

[0228] In some embodiments, a therapeutically effective amount of a compound as defined herein may be administered to a patient alone or in the form of a composition mixed with a pharmaceutically acceptable carrier, adjuvant or vehicle.

[0229] The expression "pharmaceutically acceptable carrier, adjuvant or vehicle" and equivalent expressions refer to non-toxic carriers, adjuvants or vehicles that do not destroy the pharmacological activity of the compound formulated therewith. Pharmaceutically acceptable carriers, adjuvants or vehicles that can be used in the compositions of the present invention include, but are not limited to, ion exchangers, aluminum oxide, aluminum stearate, lecithin, serum proteins (e.g., human serum albumin), buffer substances (e.g., 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 silicon dioxide, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and lanolin.

[0230] The compositions described herein can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally or via an implanted reservoir. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. Other modes of administration also include intradermal or transdermal administration.

[0231] 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, the liquid dosage form may also contain inert diluents commonly used in the art, such as water or other solvents, solubilizers, and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oil (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and sorbitan fatty acid esters and mixtures thereof. In addition to inert diluents, oral compositions may also include adjuvants, such as wetting agents, emulsifiers and suspending agents, surfactants, sweeteners, flavoring agents, and aromatics.

[0232] Injectable preparations, such as sterile injectable aqueous or oily suspensions, can be prepared using suitable dispersants or wetting agents and suspending agents according to known techniques. Sterile injectable preparations can also be sterile injectable solutions, suspensions or emulsions in nontoxic parenteral acceptable diluents or solvents, such as solutions in 1,3-butanediol. Acceptable vehicles and solvents that can be used include water, Ringer's solution, USP and isotonic sodium chloride solution. In addition, sterile, non-volatile oils are generally used as solvents or suspending media. For this purpose, any mild non-volatile oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid are also used to prepare injections.

[0233] The 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 that can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0234] In order to prolong the effect of the compound provided, it is usually desired to slow down the absorption of the compound injected subcutaneously or intramuscularly. This can be achieved by using a liquid suspension of a crystalline or amorphous material with poor water solubility. The absorption rate of the compound depends on its dissolution rate, and the dissolution rate can depend on crystal size and crystalline form. Alternatively, the delayed absorption of the compound form for parenteral administration is achieved by dissolving or suspending the compound in an oil vehicle. The injectable reservoir form is prepared by forming a microcapsule matrix of the compound in a biodegradable polymer (e.g., polylactide-polyglycolide). According to the ratio of the compound to the polymer and the property of the specific polymer used, the rate at which the compound is released can be controlled.

[0235] Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.

[0236] Compositions for rectal administration are preferably suppositories which can be prepared by mixing a compound of the present description 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.

[0237] 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 starches, lactose, sucrose, glucose, mannitol and / or silicic acid, b) binders such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone (PVP), sucrose and acacia, c) humectants such as glycerol, d) disintegrants such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate, e) solution retarder such as paraffin, f) absorption promoters such as quaternary ammonium compounds, g) wetting agents such as cetyl alcohol and glyceryl monostearate, h) absorbents such as kaolin and bentonite, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also contain a buffer.

[0238] Solid compositions of similar type can also be used as fillers in soft and hard filled gelatin capsules, using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols, etc. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells, such as enteric coatings and other coatings known in the field of pharmaceutical formulation. They can optionally contain opacifiers and can also have such a composition: they only or preferably release one or more active ingredients in a delayed manner in a specific part of the intestinal tract. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of similar type can also be used as fillers in soft and hard filled gelatin capsules, using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols, etc.

[0239] The composition can also be in the form of a microcapsule with one or more excipients as described above. The solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells, such as enteric coatings, controlled release coatings and other coatings known in the field of pharmaceutical formulation. In such solid dosage forms, the active compound can be mixed with at least one inert diluent such as sucrose, lactose or starch. According to normal practice, such dosage forms can also include other substances except 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 form can also include a buffer. They can optionally contain an opacifier and can also have such a composition: they only or preferably release one or more active ingredients in a delayed manner in a specific part of the intestinal tract. Examples of embedding compositions that can be used include polymeric substances and waxes.

[0240] The dosage form of the compound for topical or transdermal administration of this specification includes ointment, paste, cream, lotion, gel, powder, solution, spray, inhalant or patch. Active ingredient is mixed with pharmaceutically acceptable carrier and any required preservative or buffer that may be needed under aseptic conditions. Ophthalmic preparations, ear drops and eye drops are also included in the scope of this specification. In addition, this specification considers the use of transdermal patch, which has the additional advantage of providing controlled delivery of compound to health. Such dosage form can be prepared by dissolving or dispersing the compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound through the skin. Rate can be controlled by providing a rate control membrane or by dispersing the compound in a polymer matrix or gel.

[0241] The pharmaceutically acceptable compositions provided herein can also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the art of pharmaceutical formulation and can be prepared as saline solutions using benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons and / or other conventional solubilizing or dispersing agents.

[0242] The pharmaceutically acceptable compositions provided herein can be formulated for oral administration. Such products can be administered with or without food. In some embodiments, the pharmaceutically acceptable compositions of the present invention are administered without food. In other embodiments, the pharmaceutically acceptable compositions of the present invention are administered with food.

[0243] The amount of compound that can be combined with the carrier materials to produce a composition in a single dosage form will vary depending on the patient to be treated and the particular mode of administration. Provided compositions can be formulated so that a dosage of between 0.01 mg / kg body weight / day and 100 mg / kg body weight / day of the inhibitor can be administered to a patient receiving these compositions.

[0244] 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, weight, general health, sex, diet, time of administration, rate of excretion, drug combination, the judgment of the treating physician, and the severity of the symptoms associated with the proliferative disease or disorder. The amount of the compound provided in the composition will also depend on the specific compound in the composition.

[0245] The compounds or compositions described herein can be administered using any amount and any route of administration that is effective in treating symptoms as contemplated herein or in alleviating the severity of symptoms as contemplated herein. The exact amount required varies from subject to subject, depending on the species, age and general condition of the subject, the severity of the infection, the specific agent, its mode of administration, etc. The provided compounds are preferably formulated as unit dosage forms for ease of administration and uniformity of dosage. The expression "unit dosage form" as used herein refers to a physically discrete dosage unit suitable for a patient to be treated. However, it should be understood that the total daily dosage of the compounds and compositions of the present invention will be determined by the attending physician within the scope of reasonable medical judgment.

[0246] The pharmaceutically acceptable compositions of the present invention can be administered to humans and other animals orally, rectally, parenterally, intracisternal, intraperitoneally, topically (e.g., by powders, ointments, or drops), buccally, as an oral or nasal spray, etc., depending on the severity of the infection being treated. In certain embodiments, the provided compounds can be administered orally or parenterally at a dosage level 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, once or more a day to obtain the desired therapeutic effect.

[0247] It should be understood that the total daily dosage of the compound and composition of the present invention will be determined by the attending physician within the scope of reasonable medical judgment. The total daily inhibitory dose of the compound of the present invention administered to an object in a single or divided dose can be, for example, 0.01mg / kg body weight to 50mg / kg body weight or more generally 0.1mg / kg body weight to 25mg / kg body weight. Single-dose compositions can contain such an amount or its approximate number to constitute a daily dose. In one embodiment, the treatment regimen according to the present invention includes applying one or more compounds of the present invention of about 10mg to about 1000mg to the patient of such treatment in need of a single dose or multiple dose every day.

[0248] Depending on the disease or condition to be treated, additional therapeutic agents may also be present in the compositions of the invention or administered separately as part of a dosage regimen, such as additional chemotherapeutic agents. Non-limiting examples of additional therapeutic agents that can be used in combination with the compounds of the invention include anti-proliferative compounds, such as aromatase inhibitors; antiestrogens; antiandrogens; gonadorelin agonists; topoisomerase I inhibitors; topoisomerase II inhibitors; microtubule active agents; alkylating agents; retinoids, carotenoids, tocopherols; cyclooxygenase inhibitors; MMP inhibitors; antimetabolites; platinum compounds; methionine aminopeptidase inhibitors; bisphosphonates; anti-proliferative antibodies; heparanase inhibitors; inhibitors of Ras oncogenic isoforms; telomerase inhibitors; proteasome inhibitors; compounds for the treatment of hematological malignancies; kinesin spindle protein inhibitors; Hsp90 inhibitors; mTOR inhibitors; PI3K inhibitors; Flt-3 inhibitors; CDK4 / 6 inhibitors; HER2 inhibitors (Herceptin, trastuzumab); EGFR inhibitors (Iressa, Tarceva, Helian, Tagrisso, Erbitux); RAS inhibitors; MEK inhibitors (trametinib, bemetinib, cobimetinib); ERK inhibitors (Uritinib); anti-PD-1 antibodies (Opdivo, Keytruda); anti-CTLA4 antibodies (Yervoy); anti-tumor antibiotics; nitrosoureas; compounds that target / reduce the activity of protein or lipid kinases, compounds that target / reduce the activity of protein or lipid phosphatases, or any other anti-angiogenic compounds.

[0249] The treatment may also be supplemented with other treatments or interventions, such as surgery, radiation therapy (e.g., gamma radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, brachytherapy, and systemic radioisotopes), biological response modifiers (e.g., interferons, interleukins, tumor necrosis factor (TNF), and drugs to mitigate adverse effects.

[0250] The recitation of an embodiment of a variable herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof. The recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof.

[0251] Example

[0252] List of abbreviations:

[0253] Ac:Acetyl

[0254] AcOEt or EtOAc: Ethyl acetate

[0255] AcOH: acetic acid

[0256] Ar:Aryl

[0257] ATCC: American Type Culture Collection

[0258] ATP: Adenosine Triphosphate

[0259] BINOL: [1,1'-Binaphthalene]-2,2'-diol

[0260] Boc: tert-Butyloxycarbonyl

[0261] BOP: (Benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate

[0262] br: broad

[0263] BSA: Bovine Serum Albumin

[0264] CCL: Cancer Cell Line

[0265] CDCl 3 : Chloroform-d

[0266] DCE: 1,2-Dichloroethane

[0267] DCM: Dichloromethane

[0268] DIEA (or DIPEA): N,N-Diisopropylethylamine (Huenig's base)

[0269] DME: 1,2-Dimethoxyethane

[0270] DMF: N,N-Dimethylformamide

[0271] DMSO: Dimethyl Sulfoxide

[0272] DMSO-d 6 : Dimethyl Sulfoxide-d

[0273] DTT: Dithiothreitol

[0274] EA: Ethyl Acetate

[0275] EC 50 : Half Maximal Effective Concentration

[0276] ECL: Enhanced Chemiluminescence

[0277] EDTA: Ethylenediaminetetraacetic Acid

[0278] Et 2 O: Diethyl Ether

[0279] EtOH: Ethanol

[0280] Eu: Europium

[0281] FBS: Fetal Bovine Serum

[0282] GST: Glutathione S-transferase

[0283] HATU: O-(7-azabenzotriazol-1-yl)-N,N,N',N',-tetramethyluronium hexafluorophosphate

[0284] HEPES: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

[0285] Het:Heterocyclic

[0286] Hex: Hexane

[0287] HRMS: High Resolution Mass Spectrometry

[0288] HPLC: High Performance Liquid Chromatography

[0289] HRP: Horseradish peroxidase

[0290] IC 50 : Half maximal inhibitory concentration

[0291] IPA: Isopropyl alcohol

[0292] iPrOH: isopropyl alcohol

[0293] LCMS: Liquid chromatography mass spectrometry

[0294] MeCN: Acetonitrile

[0295] MS: Mass spectrometry

[0296] NMP: N-methylpyrrolidone

[0297] NMR: Nuclear Magnetic Resonance

[0298] ON: Overnight

[0299] PBS: Phosphate buffered saline

[0300] pERK: phosphorylated extracellular signal-regulated kinase

[0301] PMB: p-methoxybenzyl

[0302] PMSF: Phenylmethylsulfonyl fluoride

[0303] Rf: retention factor

[0304] RPMI-1640: Roswell Park Memorial Institute Medium

[0305] RT or rt: room temperature

[0306] SDS: Sodium dodecyl sulfate

[0307] SDS-PAGE: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

[0308] SEM: trimethylsilylethoxymethyl

[0309] SNAr: Nucleophilic Aromatic Substitution

[0310] TBST: Tris-buffered saline with 0.2% Tween-20

[0311] TBTU: O-(Benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate

[0312] TEV: Tobacco Etch Virus Protease

[0313] TFA: trifluoroacetic acid

[0314] THF: Tetrahydrofuran

[0315] TLC: Thin layer chromatography on silica gel

[0316] Ts: p-Toluenesulfonate

[0317] Y MIN : Minimum data point of the dose-activity curve

[0318] 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 drawings.

[0319] The examples presented below provide the synthesis and experimental results obtained for certain exemplary compounds. As is well known to those skilled in the art, the reaction is carried out in an inert atmosphere (nitrogen or argon) when necessary to protect the reaction components from air and moisture. Temperatures are given in degrees Celsius (° C.). Unless otherwise stated, solution percentages and ratios represent volume to volume relationships. The reactants used in the following examples can be obtained as described herein, or if not described herein, they themselves can be commercially available or can be prepared from commercially available materials by methods known in the art. The reaction was performed using a Teledyne Isco Rf Combiflash instrument on silica (SiO 2 ) at 254 nm using commercial normal phase silica. Mass spectrometric analysis was recorded using electrospray mass spectrometry. NMR was recorded on a 400 MHz Varian instrument or a 600 MHz Bruker instrument.

[0320] Preparative HPLC was performed using an Agilent instrument using a Phenomenex-Kinetex C18 (21x100 mm, 5μm) column with a flow rate of 20 mL / min (RT) and UV detection at 220 nm and 254 nm. Unless otherwise specified, 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). As noted in the text, 0.05% TFA or 0.1% AcOH or other additives were occasionally used in place of 0.1% formic acid as additives in both solvents. MeCN was also used in place of MeOH in both mobile phases to achieve more challenging separations as specified in the text. Specific gradient conditions are provided in the Examples, but the following are representative: T(0) → T(3min) isocratic, using 10% to 50% solvent B, depending on compound polarity, followed by a 12-minute gradient to 100% solvent B. The last 5 minutes were filled with 100% solvent B.

[0321] LCMS analysis was performed on an Agilent instrument. Liquid chromatography was performed on a 3×30 mm column at a flow rate of 1.5 mL / min (RT) and UV detection at 220 nm and 254 nm. The mobile phase consisted of solvent A (95% H 2 O / 5% MeOH / 0.1% AcOH) and solution B (95% MeOH / 5% H 2 O / 0.1% AcOH) with the following gradient: T(0) 100% A→T(0.5 min) 100% B→isocratic 100% B to T(2 min). MS detection was performed in parallel in both positive and negative modes using APCI detection.

[0322] Unless otherwise stated, all numerals of the amount, reaction conditions, concentration, property, stability, etc., of the expression components used in the specification and claims should be understood to be modified by the term "about" in all cases. At least, each numerical parameter should at least be interpreted according to the number of significant figures reported and by applying common rounding techniques. Therefore, unless otherwise indicated, the numerical parameters proposed in this specification and the appended claims are approximate values, which can be changed according to the properties sought to be obtained. Although the numerical range and parameters of the wide range of embodiments are approximate values, the numerical values ​​set forth in the specific embodiments are reported as accurately as possible. However, any numerical value essentially contains certain errors caused by the changes in experiments, test measurements, statistical analysis, etc.

[0323] Synthesis, biological activity and characterization of examples:

[0324] All compounds as defined herein were prepared according to the methods shown in Tables 3 and 4. Characterization data obtained by mass spectroscopy and NMR are provided for each example. The compounds were tested in the assays described in the Biological Experiments section. The conventions for reporting biological data are provided as footnotes in the various tables.

[0325] Synthesis method A:

[0326] Commercially available 2,6-difluoroaniline A-1 can be converted to its acetanilide A-2 using an acetylating agent such as acetic anhydride, and to mono-protected diphenylamine A-3, as described in WO 2012 / 101238A1. Sulfonylation to sulfonamide A-4 can be achieved using a sulfonylating agent (e.g., sulfonyl chloride) in the presence of an organic base (e.g., pyridine) with or without a catalyst (e.g., 4-dimethylaminopyridine) and a solvent (e.g., dichloromethane or tetrahydrofuran). Acetanilide A-4 is treated with aqueous hydrochloric acid in the presence of a cosolvent such as an alcohol to give an aniline salt A-5.

[0327] Commercially available 2-amino-2-cyanoacetamide A-6 can be converted to 5-amino-2-(methylthio)thiazole-4-carboxamide A-8 according to the 2-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. Following the procedure also described in Indian J. Chem., 2010, 49B, 1229, subsequent treatment with a chlorinating agent (e.g., thionyl chloride or phosphoryl chloride) in the presence of a catalytic amount of DMF provides the chloro compound A-10.

[0328] Intermediate A-11 can be obtained by heating chloropyrimidine A-10 with aniline salt A-5 in an organic acid (e.g. acetic acid). Inhibitors of general formula WI are prepared from intermediates of general formula A-11 by a two-step procedure, which involves first oxidizing the thiomethyl group generally to a mixture of the corresponding methyl sulfoxide and methyl sulfone, which is then reacted with a nucleophile (e.g. 1° amine or 2° amine, or NH-containing heterocycle, etc.). The latter step is generally carried out in the presence of a base (e.g. an organic base such as DIEA, trimethylamine, pyridine, etc., or an inorganic base such as potassium carbonate, cesium carbonate, and sodium carbonate), in a solvent (e.g. DMSO or NMP), at a temperature range of 70°C to 140°C.

[0329] Synthesis Method A

[0330]

[0331] Synthesis method B:

[0332] Inhibitors of general formula W-II are prepared as described in Synthesis Method B. Intermediates of general formula A-11 are first prepared according to Synthesis Method A and then oxidized to a mixture of methyl sulfoxide and methyl sulfone as described in Synthesis Method A. They are then coupled to 3-indolecarboxylates (e.g., methyl esters, X=CH) or 3-indazolecarboxylates (e.g., methyl esters, X=N) according to similar protocols as described for the introduction of nucleophiles in Synthesis Method A. The latter step is usually carried out in the presence of a base (e.g., an organic or inorganic base, such as Cs 2 CO 3 The ester protecting group is deprotected 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 a temperature range of 60°C to 140°C, in the presence of 2,4-dimethylamine, 6,6-dioxane, 1,2 ... 4 Cl aqueous solution or KHSO 4 The intermediate B-1 or B-2 is acidified with an aqueous solution of 1% hydroxyl group (e.g., 4-hydroxyl group ...

[0333] Synthesis Method B

[0334]

[0335] Synthesis method C:

[0336] Commercially available bromobenzimidazole C-1 can be cross-coupled with heteroarylboronic acid or boronate ester under palladium-catalyzed Suzuki-Miyaura cross-coupling conditions in the presence of a base such as sodium carbonate or potassium carbonate in a solvent such as dioxane or dimethoxyethane and water to provide intermediate C-2. Substituted benzimidazole derivatives C-2 can then be attached to intermediate A-11 after oxidation of the methyl sulfide moiety as described in the general scheme of method A to provide inhibitors of general structure W-III.

[0337] Synthesis Method C

[0338]

[0339] Synthesis method D:

[0340] Unprotected 3-indolesulfonyl chloride D-2 can be prepared as described in Org.Lett.2011,13,3588. This reagent can then be converted to the corresponding sulfonamide D-3 by reaction with a 1° amine or a 2° amine in the presence of an organic base such as DIEA or triethylamine. The D-3 fragment can then be attached to the intermediate A-11 after oxidation of the methyl sulfide moiety as described in the general scheme of method A to provide inhibitors of general structure W-IV.

[0341] Synthesis Method D

[0342]

[0343] Synthesis method E:

[0344] Alternatively, inhibitors of general structure W-IV can be obtained by 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 is then converted to the corresponding sulfonamide E-2 by reaction with a 1° amine or a 2° 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 be attached to the intermediate A-11 after oxidation of the methyl sulfide moiety as described previously.

[0345] Synthesis Method E

[0346]

[0347] Synthesis method F:

[0348] 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 nonahydrate and directly alkylated without separation from the alkyl halide to provide the sulfide intermediate F-2. The sulfide intermediate F-2 is then converted to the sulfone intermediate F-3 using an oxidizing agent such as 3-chloroperoxybenzoic acid. The final inhibitor of general structure WV is then obtained by connecting the indole sulfone F-3 with the intermediate A-11 after oxidation of the methyl sulfide moiety as described above.

[0349] Synthesis Method F

[0350]

[0351] Synthesis method G:

[0352] Bright red solutions of commercially available 3-fluoro-2-nitroanilines G-1 react with primary or secondary amines in solvents such as MeCN, DMSO, NMP or DMF and in the presence of an inorganic base such as potassium carbonate or an organic base such as DIEA, heated under thermal or microwave conditions in the temperature range of 40°C to 120°C to provide intermediates G-2. Reduction of the nitro group of intermediate G-2 and subsequent cyclization of the resulting 1,2-phenylenediamine intermediate to the desired benzimidazole intermediate G-3 can be achieved in a single operation using iron metal, ammonium chloride and formic acid in an alcohol solvent such as isopropanol, heated in the temperature range of 40°C to 90°C. The final inhibitors of general structure W-VI are then obtained by heating the substituted benzimidazole G-3 and intermediate A-11, which has been oxidized to the corresponding methyl sulfone / methyl sulfoxide under conventional conditions as described above, in the presence of a base.

[0353] Synthesis Method G

[0354]

[0355] Procedures for preparing inhibitors which do not or only partially fall within the general synthetic procedure described above are described in detail below.

[0356] Sulfonyl chloride:

[0357] 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.

[0358] Other sulfonyl chlorides were prepared by using or adapting literature procedures as described below.

[0359] 3-Fluoro-2-methyl-4-methoxybenzenesulfonyl chloride:

[0360]

[0361] 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 (2 x 10 mL acetone for rinsing) 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 and washed with Na 2 SO 4Dry, filter and concentrate under reduced pressure. The residue was purified by flash chromatography on silica gel using 0 to 5% EtOAc / hexanes to give 2-fluoro-3-methylanisole as a clear colorless liquid (5.30 g, 95% yield): 1 H NMR (CDCl 3 )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).

[0362] Step 2: To a solution of 2-fluoro-3-methylanisole (1.00 g, 7.13 mmol) in DCM (5.6 mL) from step 1 was added a solution of chlorosulfonic acid (1.13 mL, 16.5 mmol) in DCM (5.6 mL) over a period of 5 minutes. The light brown reaction mixture containing a viscous liquid layer was stirred at room temperature for 10 minutes and then quenched by pouring into a mixture of water (10 mL) and ice (5 g). The aqueous phase was extracted with DCM (2×10 mL) and heated to 40° C. for 10 minutes. The resulting mixture was stirred for 10 minutes at room temperature. The mixture was stirred for 10 minutes at room temperature for 10 minutes. The reaction mixture was stirred for 2 ... 2 SO 4 Drying over high temperature, filtering and concentrating under reduced pressure gave the desired sulfonyl chloride as a colorless liquid (1.70 g, 100% yield). The material was used without further purification: 1 H NMR (CDCl 3 )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).

[0363] 3-Chloro-2-methyl-4-methoxybenzenesulfonyl chloride:

[0364]

[0365] Following an analogous procedure to 3-fluoro-2-methyl-4-methoxybenzenesulfonyl chloride (step 1) and starting from 2-chloro-3-methylphenol, 2-chloro-3-methylanisole was obtained as a colorless liquid in quantitative yield: 1 H NMR (CDCl 3 )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).

[0366] Treatment with chlorosulfonic acid as described for 3-fluoro-2-methyl-4-methoxybenzenesulfonyl chloride (step 2) provided 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).

[0367] General synthetic method A: Preparation of difluoroaniline hydrochloride intermediate A-5 (Ar = 2,3-dichlorophenyl) from acetanilide A-3.

[0368]

[0369] Preparation of acetanilide A-2: Acetanilide A-2 can be prepared by acetylation of 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 aniline as described in WO 2012 / 101238A1.

[0370] 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 eq., 14.7 mL) was added to the brown solution, followed by 2,3-dichlorobenzenesulfonyl chloride (1.2 eq., 13.45 g). The resulting reaction mixture was stirred at 45 °C for 3.5 hours, after which the conversion was complete as judged by LCMS monitoring. The reaction mixture was cooled to room temperature and then partitioned between EtOAc and 2-Me-THF (1:1) and water. 1N HCl solution was added until a slightly acidic pH was obtained. A large amount of off-white solid was present in the biphasic mixture and was filtered off (first batch). The filtrate layers were separated and the aqueous layer was extracted twice more with EtOAc. The combined organic extracts were washed once with water, then with brine, and filtered through MgSO 4 Dried, filtered and concentrated to ~20 mL. The resulting suspension was sonicated, and the solid was collected by filtration and washed with EtOH (Batch 2). The two batches were combined and dried under reduced pressure. A-4 (15.3 g, 85% yield) was obtained as a beige solid and used without further purification: 1 H NMR (DMSO-d 6 )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 + ).

[0371] 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 equipped with a reflux condenser with a stopper and heated at 80 °C with stirring. After 24 hours, the conversion was judged to be ~70% according to LCMS monitoring. Another portion of EtOH (65 mL) and 6N HCl (65 mL) were added to the suspension and stirring was continued at 80 °C for more than 7 hours, after which LCMS indicated complete conversion to the desired aniline. The reaction mixture was diluted with 50 mL of water while hot 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 3 times under reduced pressure and then dried under vacuum to give 7.2 g of the desired product A-5 in the form of the HCl salt as a yellow solid: 1 H NMR(DMSO-d 6 ) δ: 10.30 (s, 1H), 7.93 (dd, J = 8.2, 1.2 Hz, 1H), 7.83 (dd, J = 8.0, 1.4 Hz, 1H), 7.49 (t, J = 8.0 Hz, 1H), 6.68 - 6.96 (m, 1H), 6.31 (td, J = 8.6, 5.5 Hz, 1H). MS m / z 350.9 (M-H).

[0372] The following intermediate A-5 (Table 1) was prepared in a similar manner using the relevant sulfonyl chloride:

[0373] Table 1

[0374]

[0375] General synthetic method A - Preparation of inhibitor W-I from intermediate A-5

[0376]

[0377] 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.

[0378] Step 3: Preparation of intermediate A-9 (adapted from Indian J. Chem., 2010, 49B, 1229): 5-amino-2-methylsulfonyl-thiazole-4-carboxamide (3.19 g, 16.855 mmol) was evenly divided into two microwaveable 20 mL vials, and formamide (13.4 mL, 337 mmol) was added to each vial. The vials were sealed and heated in a microwave at 185° C. for 20 minutes each, and then heated at 190° C. for another 15 minutes. The mixture was 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 some cold water. After suction drying on the filter, the solid was transferred to a vial and dried under reduced pressure. 2-Methylsulfonyl-6,7a-dihydro-3aH-thiazolo[5,4-d]pyrimidin-7-one (2.34 g, 69% yield) was obtained as a yellow to beige solid. MS m / z 202.2 (MH + ). 1 H NMR (DMSO-d 6 )δ:12.81(br.s.,1H),8.15(s,1H),2.75(s,3H).

[0379] Step 4: Preparation of intermediate A-10 (adapted from Indian J. Chem., 2010, 49B, 1229): 2-methylsulfonyl-6H-thiazolo[5,4-d]pyrimidin-7-one (1.00 g, 5.02 mmol) was suspended in thionyl chloride (10 mL, 137 mmol), and 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 had become a clear solution. Cooled to room temperature, then diluted with some toluene and concentrated to dryness under reduced pressure. The residue was resuspended in toluene and concentrated to dryness again. The product was then dried under reduced pressure. 7-Chloro-2-methylsulfonyl-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).

[0380] Step 5: Preparation of intermediate A-11 (Ar = 2,3-dichlorophenyl): N-(3-amino-2,4-difluoro-phenyl)-2,3-dichloro-benzenesulfonamide; The hydrochloride (811 mg, 2.08 mmol) and 7-chloro-2-methylsulfonyl-thiazole [5,4-d] pyrimidine (544 mg, 2.50 mmol) were charged into 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 ensure homogeneity, and the solid was then collected by filtration and washed with a little water. The solid was drained on the filter and then further dried under reduced pressure. 2,3-Dichloro-N-[2,4-difluoro-3-[(2-methylsulfonylthiazolo[5,4-d]pyrimidin-7-yl)amino]phenyl]benzenesulfonamide (1.17 g, 100% yield) was obtained as a yellow solid and used without further purification. MS m / z 534.0 (MH + ). 1 H NMR (DMSO-d 6 )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, 1H), 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).

[0381] The following intermediates A-11 were prepared using a similar sequence as described (Table 2):

[0382] Table 2

[0383]

[0384] Step 6: Preparation of methyl sulfone and methyl sulfoxide mixture from intermediate A-11 (Ar = 2,3-dichlorophenyl): 2,3-Dichloro-N-[2,4-difluoro-3-[(2-methylsulfonylthiazolo[5,4-d]pyrimidin-7-yl)amino]phenyl]benzenesulfonamide (1.00 g, 1.87 mmol) was suspended in DCM (25 mL). Then 3-chloroperoxybenzoic acid (839 mg, 3.74 mmol) was added in one portion. The mixture was stirred at room temperature for 23 hours. A saturated solution of sodium thiosulfate (~1 mL) was added and the mixture was stirred vigorously for 10 minutes. It was then diluted with DCM and washed with NaHCO 3The mixture was washed three times with a 1:1 mixture of a saturated solution of DCM and water. Acetic acid was slowly added to neutralize the aqueous layer (pH 7). The aqueous layer was then back-extracted twice with DCM, and the combined organic layers were then washed once with water and then with brine. Na 2 SO 4 The organic layer was dried, filtered, concentrated and dried under reduced pressure. The resulting beige solid was adsorbed onto and silica gel, then through a 25 g silica gel column, using a 5% to 100% EtOAc in hexane gradient to purify by flash chromatography. Two major peaks were separated:

[0385] Methyl sulfone: rf = 0.72 @ 8:2 EtOAc / hexanes, 710 mg (67% yield), as a beige solid. MS m / z 566.2 (MH + ). 1 H NMR (DMSO-d 6 )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).

[0386] Methyl sulfoxide: rf = 0.31 @ 8:2 EtOAc / hexanes, 229.1 mg (22% yield), as a beige solid. MS m / z 550.2 (MH + ). 1 H NMR (DMSO-d 6 )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).

[0387] Note: The subsequent reaction (ie, step 7 of general synthetic method A) does not require separation and purification of the two products by chromatography, and they can be used as a crude mixture.

[0388] 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 and a 0.1% formic acid modifier. The appropriate fractions were combined and concentrated. The residue was lyophilized from a mixture of water and MeCN. 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) was obtained as a beige solid.

[0389] Additional examples of inhibitors prepared in a similar manner are listed in Table 3 under Method A.

[0390] General Method for Synthesis of Inhibitors of Formula W-II (Method B, X=CH) - Synthesis of Example 28

[0391]

[0392] Step 1: Preparation of methyl sulfone and methyl sulfoxide mixture from intermediate A-11 (Ar = 2-methyl-3-fluorophenyl): N-[2,4-difluoro-3-[(2-methylsulfonylthiazolo[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 (~0.2 mL) was added and the mixture was stirred vigorously for 10 minutes. It was then diluted with DCM and washed with NaHCO 3 The saturated solution and water were washed three times with a 1:1 mixture. The combined aqueous layers were back extracted once with DCM, and the combined organic layers were washed with water and then with brine. 2 SO 4The organic layer was dried, filtered, concentrated and dried under reduced pressure. The resulting beige solid (445.5 mg, 93% yield, 2:1 mixture of sulfone and sulfoxide) was used as is without purification. MS m / z 530.2 and 514.2 (MH + ).

[0393] Step 2 and step 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 loaded into 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.

[0394] The reaction mixture was cooled to room temperature and 4N sodium hydroxide solution (0.85mL, 3.40mmol) was added. The resulting mixture was stirred at 50°C for another hour. The reaction mixture was cooled to room temperature and then transferred to 100mL 1N HCl. After the resulting suspension was sonicated, the solid was 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 (292mg, 84.3% yield) as a brown solid. The crude material was used without further purification in the next step. MS m / z 611.2 (MH + ).

[0395] 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) and then dissolved in 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)ethyl-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 with 0.5 mL of formic acid, diluted with a little DMSO, and then purified by preparative reverse phase HPLC to give the compound of Example 28 (9.8 mg, 23% yield) as a beige solid after freeze drying from a MeCN / water mixture.

[0396] Additional examples of inhibitors prepared in a similar manner are listed in Table 3 under Method B.

[0397] General Method for Synthesis of Inhibitors of Formula W-II (Method B, X=N) - Synthesis of Example 23

[0398]

[0399] Step 1: Prepare the methyl sulfone and methyl sulfoxide mixture from intermediate A-11 as described above in Example 28 (Step 1).

[0400] Step 2 and step 3: preparation of carboxylic acid B-2: The crude mixture (300mg, 0.567mmol) and indazolyl-3-carboxylic acid methyl ester (150mg, 0.850mmol) from the above sulfone and sulfoxide are loaded in a 20mL vial and suspended in NMP (3.6mL). Then DIPEA (0.49mL, 2.83mmol) is added to the orange solution, and the reaction mixture is stirred at 105°C for 24h. Then cool to room temperature and add an aqueous solution of 4N sodium hydroxide (0.85mL, 3.40mmol). The resulting mixture is stirred for another 1 hour at 50°C. The reaction mixture is cooled to room temperature and then transferred to 100mL 1N HCl. After the resulting suspension was sonicated, 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 + ).

[0401] 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-pyridyl)methanamine (0.013 mL, 0.114 mmol) was added. The reaction mixture was stirred at room temperature overnight. The reaction was then quenched with 0.5 mL of formic acid, diluted with DMSO, and then purified by reverse phase preparative HPLC using a 15% to 55% MeCN water gradient and 0.1% formic acid modifier. The appropriate fractions were combined and lyophilized from the MeCN / water mixture. The compound of Example 23 was obtained as a beige solid (14 mg, 34% yield).

[0402] Additional examples of inhibitors prepared in a similar manner are listed in Table 3 under Method B.

[0403] General Method for Synthesis of Inhibitors of Formula W-III (Method C) - Synthesis of Example 8

[0404]

[0405] 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-pyridine boronic 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 was bubbled through the mixture for 1 minute, and then tetrakis(triphenylphosphine)palladium(0) (16.4 mg, 0.014 mmol) was added. Argon was bubbled through the solution again for 3 minutes, the vial was sealed and heated at 100°C for 2 hours (conversion to the desired product was complete as judged by LCMS analysis). The reaction mixture was cooled to RT, diluted with EtOAc and washed with brine. In MgSO 4 After drying on ice, the extract was concentrated under reduced pressure and purified by using Et 3 The residue was purified by flash chromatography on N-pretreated silica and DCM-20% iPrOH / DCM gradient to afford the desired benzimidazole intermediate (58 mg, 84% yield): 1 H NMR (DMSO-d 6 )δ: 12.71 (width s, 1H), 9.24 (s.1H), 8.57 (dd, J = 5.1, 1.6Hz, 1H), 8.43 (width 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 + ).

[0406] Step 2: Prepare the methyl sulfone and methyl sulfoxide mixture from intermediate A-11 as described above in Example 28 (Step 1), Method B.

[0407] Step 3: Preparation of W-III (Example 8): The substituted benzimidazole from Step 1 was coupled with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2-methyl-3-fluorophenyl) in NMP using DIPEA as described in Example 4 (Step 7) of General Method A.

[0408] Other compounds (ie, Example 9 and Example 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 in Table 3 under Method C.

[0409] General Method for Synthesis of Inhibitors of Formula W-IV (Method D) - Synthesis of Example 12

[0410]

[0411] Step 1: Prepare 3-indolesulfonyl chloride D-2 as described in Org.Lett.2011,13,3588. A solution of indole (3g, 25.6mmol) and sulfur trioxide-pyridine (4.08g, 25.6mmol) in pyridine (15mL) was heated to reflux (115°C) for 2h under stirring. After 2 hours, the reactant was cooled to room temperature and diluted with water (20mL). The aqueous layer was washed twice with diethyl ether (20mL). The aqueous layer was evaporated to dryness to obtain crude pyridine 1H-indole-3-sulfonic acid ester D-1 (5.30g, 75% yield) as a white solid. The crude material was used as such in the next step.

[0412] Step 2: Dissolve the crude pyridine 1H-indole-3-sulfonate (4.60 g, 16.7 mmol) in a 1:1 mixture of sulfolane and acetonitrile (50 mL). Cool the white suspension to 0 °C and add POCl dropwise under stirring. 3 4-Indole-3-sulfonyl chloride D-2 (740mg, 21% yield) was obtained.

[0413] Step 3: 1H-indole-3-sulfonyl chloride (0.40 g, 1.86 mmol) was charged into a 25 mL flask and 6 mL of THF was added thereto. The solution was cooled to 0 °C and 1-methylpiperazine (0.41 mL, 3.71 mmol) was then added dropwise. The mixture changed from a red solution to a light yellow solution with a large amount of sticky solids 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 (pH of the aqueous layer ~ 7) was added 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 once with brine. The mixture was then washed with MgSO 4Dry, filter and concentrate 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 sticky solid), and the liquid 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 without further purification. MS m / z 280.1 (MH + ). 1 H NMR (DMSO-d 6 )d:12.20(br.s.,1H),7.95(d,J=2.7Hz,1H),7.78(d,J=8.2Hz,1H),7.53(d,J=8.2Hz,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).

[0414] Step 4: Prepare the methyl sulfone and methyl sulfoxide mixture from intermediate A-11 as described above in Example 28 (Step 1), Method B.

[0415] Step 5: Preparation of W-IV (Example 12): The sulfonamide intermediate D-3 (3-((4-methylpiperazin-1-yl)sulfonyl)-1H-indole) was coupled with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) in NMP using DIPEA as described in Example 4 (Step 7) in General Method A.

[0416] Additional examples of inhibitors prepared in a similar manner are listed in Table 3 under Method D.

[0417] General Method for Synthesis of Inhibitors of Formula W-IV (Method E) - Synthesis of Example 70

[0418]

[0419] Step 1: Preparation of 3-indolesulfonamide fragment E-2: To a vial with a stirring bar, 1-(p-toluenesulfonyl)indole-3-sulfonyl chloride (117 mg, 0.316 mmol) (Chemical and Pharmaceutical Bulletin 2009, 57, 591) in anhydrous THF (1.5 mL) was added. The solution was cooled to 0°C and DIEA (0.11 mL, 0.633 mmol) was added dropwise. N-(2-methoxyethyl)ethylamine (0.039 mL, 0.316 mmol) was then added dropwise and the reaction mixture was slowly heated to room temperature. The reaction was stirred at room temperature for 1 h. After completion, 10% KOH aqueous solution (same volume as solvent) was added dropwise. The reaction was heated at 60°C overnight. After completion, the reaction mixture was diluted with EtOAc and NH 4 Cl aqueous solution. Separate the layers. Wash the organic layer with brine and then with MgSO 4 Dry over medium, filter and concentrate to dryness to give the expected sulfonamide derivative as a light orange oil (86 mg, 96%). MS m / z 283.2 (MH + ).

[0420] Step 2: Prepare the methyl sulfone and methyl sulfoxide mixture from intermediate A-11 as described above in Example 28 (Step 1), Method B.

[0421] Step 3: Preparation of W-IV (Example 70): The sulfonamide intermediate E-2 (N-ethyl-N-(2-methoxyethyl)-1H-indole-3-sulfonamide) is coupled with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) in DMSO using DIPEA as described in Example 4 (Step 7) in General Method A.

[0422] Additional examples of inhibitors prepared in a similar manner are listed in Table 3 under Method E.

[0423] General Method for the Synthesis of Inhibitors of Formula WV (Method F) - Synthesis of Example 63

[0424]

[0425] 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 nonahydrate (724 mg, 3.0 mmol) dissolved in water (0.5 mL), and the mixture was stirred at 50° C. for 2 hours. To the resulting mixture was then added iodoethane (0.12 mL, 1.5 mmol), 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, then with brine, and then MgSO 4 Drying, filtration and concentration in vacuo gave crude sulfide F-2 (170 mg, 95%), which was used directly in the next step without further purification. MS m / z 178.2 (MH + ).

[0426] Step 2: Preparation of indole-3-ethylsulfone (R = Et) intermediate F-3: To a solution of 3-ethylsulfonyl-1H-indole (170 mg, 0.96 mmol) in DCM (4.8 mL) was added 3-chloroperoxybenzoic acid (537 mg, 2.40 mmol) at room temperature. After stirring at this temperature for 4 hours, the mixture was diluted with EtOAc and then washed with NaHCO 3 The resulting solution was washed twice with a saturated aqueous solution of MgSO 4 The organic layer was dried, filtered, and concentrated in vacuo. The residue 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 + ).

[0427] Step 3: Prepare the methyl sulfone and methyl sulfoxide mixture from intermediate A-11 as described above in Example 28 (Step 1), Method B.

[0428] Step 4: Preparation of WV (Example 63): 3-ethylsulfonyl-1H-indole intermediate F-3 was coupled with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) in DMSO using DIPEA as described in Example 4 (Step 7) in General Procedure A.

[0429] Additional examples of inhibitors prepared in a similar manner are listed in Table 3 under Method F.

[0430] General Method for Synthesis of Inhibitors of Formula W-VI (Method G) - Synthesis of Example 113

[0431]

[0432] 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 hours. 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 + ).

[0433] 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 h. The reaction mixture was cooled to room temperature and filtered. Filter. The solution was concentrated and the crude product was purified by column chromatography (silica gel, 0-15% MeOH in DCM) to give 1-(1H-benzoimidazol-4-yl)-4-methyl-piperidin-4-ol (0.17 g, 55% yield) as a light red foam solid. MS m / z 232.2 (MH + ). 1 H NMR (400 MHz, DMSO-d 6 )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).

[0434] Step 3: Prepare the methyl sulfone and methyl sulfoxide mixture (Ar = 2,3-dichlorophenyl) from intermediate A-11 as described above in Example 28 (Step 1), Method B.

[0435] Step 4: Preparation of W-VI (Example 113): 1-(1H-Benzimidazol-4-yl)-4-methyl-piperidin-4-ol intermediate G-3 was coupled with 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 in Example 4 (Step 7) in General Method A.

[0436] Additional examples of inhibitors prepared in a similar manner are listed in Table 3 under Method G.

[0437] Preparation Example 17, Example 47 and Example 99:

[0438]

[0439] Step 1: Preparation of 4-(trifluoromethyl)-1H-benzo[d]imidazole (Example 17): In a 10 mL vial, add a solution of 3-(trifluoromethyl)benzene-1,2-diamine (280 mg, 1.59 mmol) in formic acid (2.0 mL). Heat the reaction to 100 °C and stir for 3 h. Cool the reaction to room temperature and remove the formic acid under reduced pressure. Add NaHCO to the residue. 3 The saturated solution of 2-(trifluoromethyl)-1H-benzimidazole (269 mg, 91% yield) was obtained by adding 1% ethyl acetate to a saturated solution of 1% ethyl acetate to obtain a precipitate, which was homogenized by ultrasonic treatment. 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 obtain 4-(trifluoromethyl)-1H-benzimidazole (269 mg, 91% yield) as a light brown solid. MSm / z 187.2 (MH + ). 1 H NMR (DMSO-d 6 )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).

[0440] 5,6-Dichloro-1H-benzimidazole (Example 47) was prepared in a similar manner starting from commercially available 4,5-dichloro-1,2-phenylenediamine. The product was a light 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).

[0441] 4-Methoxy-1H-benzimidazole (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 + ).

[0442] Step 2: Prepare the methyl sulfone and methyl sulfoxide mixture (Ar = 2,3-dichlorophenyl) from intermediate A-11 as described above in Example 28 (Step 1), Method B.

[0443] Step 3: Example 17, Example 47 and Example 99 were prepared by coupling the above appropriately substituted benzimidazole with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) using DIPEA in NMP as described in Example 4 in General Procedure A (Step 7).

[0444] Preparation Example 49 and Example 71:

[0445]

[0446] Step 1: Preparation of 4,5-difluoro-1H-benzimidazole (Example 49): 2,3-difluoro-6-nitro-aniline (800 mg, 4.60 mmol) was placed in a flask together with ammonium chloride (2.46 g, 46.0 mmol) and iron powder (2.57 g, 46.0 mmol). IPA (11.5 mL) was then added, followed by formic acid (12 mL, 305 mmol). The flask was equipped 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 passed through a small Filter, rinse with IPA, and concentrate the filtrate to dryness. Then use NaHCO 3 The residue was neutralized with a saturated solution of 2% MgSO and extracted three times with EtOAc. The combined organic layers were washed with brine and MgSO 4 The residue was purified by silica gel chromatography using a gradient of 50% to 100% EtOAc in DCM. The appropriate fractions were combined, concentrated and dried under reduced pressure. 4,5-difluoro-1H-benzimidazole (706 mg, 99% yield) was obtained. MS m / z 155.2 (MH + ). 1 H NMR (DMSO-d 6)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).

[0447] 4,6-Difluoro-1H-benzimidazole (Example 71) was prepared in a similar manner starting from commercially available 2,4-difluoro-6-nitro-aniline in 93% yield. MS m / z 155.2 (MH + ). 1 H NMR (DMSO-d 6 )δ: 12.87 (br.s., 1H), 8.28 (s, 1H), 7.25 (d, J = 7.8Hz, 1H), 7.05 (t, J = 10.4Hz, 1H).

[0448] Step 2: Prepare the methyl sulfone and methyl sulfoxide mixture (Ar = 2,3-dichlorophenyl) from intermediate A-11 as described above in Example 28 (Step 1), Method B.

[0449] Step 3: Example 49 and Example 71 were prepared by coupling the above appropriately substituted benzimidazole with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) using DIPEA in NMP as described in Example 4 in General Procedure A (Step 7).

[0450] Preparation Example 72:

[0451]

[0452] Step 1: Preparation of 3-methylsulfonyl-1H-indole: Indole (182 mg, 1.55 mmol) was dissolved in THF (17 mL) at room temperature under nitrogen. 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. NaHCO was added. 3 The mixture was then extracted with EtOAc. The organic layer was washed with brine and MgSO 4 Dry, filter and concentrate in vacuo.The residue was purified by silica gel chromatography using a gradient of EtOAc in hexanes (20-60%) to afford 3-methylsulfonyl-1H-indole (30 mg, 10% yield).

[0453] Step 2: Prepare the methyl sulfone and methyl sulfoxide mixture (Ar = 2,3-dichlorophenyl) from intermediate A-11 as described above in Example 28 (Step 1), Method B.

[0454] Step 3: Example 72 was prepared by coupling the above 3-methylsulfonyl-1H-indole with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) in DMSO using DIPEA as described in Example 4 in General Procedure A (Step 7).

[0455] Preparation Example 79 and Example 80:

[0456]

[0457] Step 1: Preparation of 2-fluoro-3-(2-methoxyethoxy)-6-nitro-aniline (Example 79): 2,3-difluoro-6-nitro-aniline (800 mg, 4.60 mmol) was charged into a 25 mL flask and dissolved in 10 mL DMF at room temperature to give a bright yellow solution. 2-Methoxyethanol (2.2 mL, 27.6 mmol) was then added, followed by potassium carbonate (2.54 g, 18.4 mmol). The color of the solution turned 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. The aqueous layer was extracted twice with EtOAc, the combined organic layers were washed with brine, and MgSO 4 Dry, filter and concentrate. The resulting residue is purified by silica gel chromatography using a gradient of EtOAc in hexanes. 2-Fluoro-3-(2-methoxyethoxy)-6-nitro-aniline (1.01 g, 96% yield) is obtained. 1 H NMR (DMSO-d 6 )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).

[0458] 2-(3-Amino-2-fluoro-4-nitro-phenoxy)ethanol (Example 80) was prepared in a similar manner using ethylene glycol. The yield of this product was 98%. 1 H NMR (DMSO-d 6)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).

[0459] Step 2: The substituted benzimidazoles were prepared from the 2-nitroaniline described in Step 1 using the "one pot" procedure described in Method G of Example 113 (Step 2).

[0460] 4-Fluoro-5-(2-methoxyethoxy)-1H-benzimidazole (Example 79): 82% yield. MS m / z 211.2 (MH + ). 1 H NMR (DMSO-d 6 )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).

[0461] 2-[(4-Fluoro-1H-benzimidazol-5-yl)oxy]ethanol (Example 80): 53% yield. MS m / z 197.2 (MH + ). 1 H NMR (DMSO-d 6 )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).

[0462] Step 3: Prepare the methyl sulfone and methyl sulfoxide mixture (Ar = 2,3-dichlorophenyl) from intermediate A-11 as described above in Example 28 (Step 1), Method B.

[0463] Step 4: Example 79 and Example 80 were prepared by coupling the above appropriately substituted benzimidazole with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) using DIPEA in NMP as described in Example 4 (Step 7) in General Procedure A.

[0464] Preparation Example 81:

[0465]

[0466] Step 1: Preparation of (5-chloro-1H-indol-3-yl)-(3-oxa-8-azabicyclo[3.2.1]octane-8-yl)methanone: 5-chloro-1H-indole-3-carboxylic acid (50 mg, 0.256 mmol) was dissolved in NMP (1.5 mL). 2-(1H-benzotriazole-1-yl)-1,1,3,3,-tetramethyluronium tetrafluoroborate (98 mg, 0.31 mmol) was added at room temperature, followed by 3-oxa-8-aza-bicyclo[3.2.1]octane hydrochloric acid (42 mg, 0.28 mmol). Finally, N,N-diisopropylethylamine (0.18 mL, 1.02 mmol) was 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 and then heated to MgSO 4 The residue was purified by silica gel chromatography using a gradient of 1:1 EtOAc to 100% EtOAc in DCM followed by 2% IPA in EtOAc. (5-Chloro-1H-indol-3-yl)-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)methanone (65 mg, 88% yield) was obtained. MS m / z 291.1 (MH + ). 1 H NMR (DMSO-d 6 )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).

[0467] Step 2: Prepare the methyl sulfone and methyl sulfoxide mixture (Ar = 2,3-dichlorophenyl) from intermediate A-11 as described above in Example 28 (Step 1), Method B.

[0468] Step 3: Example 81 was prepared by coupling the above (5-chloro-1H-indol-3-yl)-(3-oxa-8-azabicyclo[3.2.1]octan-8-yl)methanone with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) in DMSO using DIPEA as described in Example 4 (Step 7) in General Method A.

[0469] Preparation Example 91, Example 92, Example 106 and Example 107:

[0470]

[0471] Step 1: Preparation of 1-(1H-indol-3-yl)propan-1-one (Example 91): A 100 mL round bottom flask was charged with a solution of indole (200 mg, 1.71 mmol) in 7 mL DCM. The solution was cooled to 0°C and then 1.8 M (25 wt%) Et 2 AlCl in toluene (1.42 mL, 2.56 mmol). The reaction was stirred at 0 °C for 30 min. A solution of propionyl chloride in 6 mL DCM was added dropwise. The reaction was warmed to room temperature and stirred for 2.5 h. The reaction was quenched with 3 eq. of NaOAc in 5 mL water. The reaction was diluted with DCM and Filter to remove aluminum salts. Transfer the filtrate to a separatory funnel and add water. Add 1M NaOH to help dissolve any remaining aluminum salts. Separate the layers and wash the organic layer with water and then with brine. Then wash with MgSO 4 The organic layer was dried, filtered, and then concentrated under reduced pressure. The crude material was purified by silica gel chromatography using 100% hexanes to 75% EtOAc in a hexanes gradient. 1-(1H-indol-3-yl)propan-1-one (228.9 mg, 77% yield) was obtained as a light yellow solid. MS m / z 174.2 (MH + ). 1 H NMR (400 MHz, DMSO-d 6 )δ:11.88(br.s.,1H),8.30(s,1H),8.15-8.23(m,1H),7.41-7.49(m,1H),7.18(qu int,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).

[0472] 1-(1H-Indol-3-yl)-2-methyl-propan-1-one (Example 92) was prepared in a similar manner using isobutyric acid chloride. The yield of the product was 94%. MS m / z 188.2 (MH + ). 1 H NMR (400 MHz, DMSO-d 6 )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).

[0473] 1-(1H-indol-3-yl)-3-methoxy-propan-1-one (Example 106) was prepared in a similar manner using 3-methoxypropanoyl chloride. The yield of the product was 86%. MS m / z 204.2 (MH + ).

[0474] 1H-Indol-3-yl(2-thienyl)methanone (Example 107) was prepared in a similar manner using 2-thiophenecarbonyl chloride. The yield of the product was 80%. MS m / z 228.0 (MH + ).

[0475] Step 2: Prepare the methyl sulfone and methyl sulfoxide mixture (Ar = 2,3-dichlorophenyl) from intermediate A-11 as described above in Example 28 (Step 1), Method B.

[0476] Step 3: Example 91, Example 92, Example 106 and Example 107 were prepared by coupling the above appropriately substituted indole with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) using DIPEA in DMSO as described in Example 4 in General Procedure A (Step 7).

[0477] Preparation Example 98:

[0478]

[0479] 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 charged into a 25 mL flask and suspended in 10 mL 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 stirred at 40 ° C for 4 hours. The reaction mixture was diluted with EtOAc. The solid was removed by filtration and the filtrate was concentrated to dryness. The resulting residue was purified by silica gel flash chromatography using a hexane gradient of 0% to 40% EtOAc. The appropriate fractions were 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).

[0480] Step 2: tert-Butyl 3-(2-benzyloxyethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (673 mg, 1.94 mmol) was charged into a 100 mL flask and dissolved in 5 mL dioxane. A solution of 4N HCl in dioxane (1.94 mL, 7.76 mmol) was then added dropwise. After stirring for 1 hour, another portion of HCl solution (2.91 mL, 11.64 mmol) was added and the mixture was heated to 40°C. After stirring for another 2 hours at this temperature, a colloid 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, concentrated to dryness again, and then dried under reduced pressure. 3-(2-benzyloxyethyl)-3,8-diazabicyclo[3.2.1]octane; dihydrochloride (620 mg, 100% yield) was obtained as a white sticky solid and was used as such. MS m / z 247.3 (MH + ). 1H 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 is hidden under the peak of water).

[0481] Step 3: 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 in a 100 mL flask was added 10% palladium on charcoal (205 mg, 0.193 mmol). The flask was placed under reduced pressure and then filled with hydrogen. This operation was repeated 3 more times and the mixture was then stirred vigorously under a balloon atmosphere of hydrogen over the weekend. The flask was flushed with nitrogen and passed through a short block. The reaction mixture was filtered. The filtrate was concentrated and dried under reduced pressure. 2-(3,8-diazabicyclo[3.2.1]octan-3-yl)ethanol; dihydrochloride (406 mg, 92% yield) was obtained. 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) (2H is hidden under the peak of water).

[0482] Step 4: Example 98 was prepared by coupling the above amine to intermediate B-1 (Ar = 2,3-dichlorophenyl) in a similar manner as described in General Procedure B (Step 4) of Example 28 by using TBTU instead of HATU.

[0483] Preparation Example 100:

[0484]

[0485] Step 1: Preparation of 6-bromo-4-fluoro-1H-benzimidazole: This compound was prepared starting from commercially available 4-bromo-2-fluoro-6-nitro-aniline using iron, ammonium chloride and formic acid in a similar manner to the benzimidazole described in Example 49 / 71 (Step 1). 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 + ). 1 H NMR (DMSO-d6) δ: 12.94 (br.s., 1H), 8.31 (s, 1H), 7.63 (s, 1H), 7.28 (d, J = 10.6Hz, 1H).

[0486] Step 2: Preparation of 2-[(6-bromo-4-fluoro-benzoimidazol-1-yl)methoxy]ethyl-trimethyl-silane and 2-[(5-bromo-7-fluoro-benzoimidazol-1-yl)methoxy]ethyl-trimethyl-silane: 6-bromo-4-fluoro-1H-benzimidazole (500 mg, 2.05 mmol) was dissolved in 10 mL of DMF at room temperature, and potassium carbonate (848 mg, 6.14 mmol) was then added, followed by SEM-Cl (0.44 mL, 2.46 mmol). The mixture was stirred at the same temperature for 18 hours. Another 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 3 times with EtOAc. The combined organic layers were washed with water, then with brine, and then with MgSO 4 Dried, filtered and concentrated. The resulting residue was purified by silica gel chromatography using a hexane gradient of 100% hexane to 40% EtOAc. The appropriate fractions were combined, concentrated and dried under reduced pressure. An approximately 3:1 mixture of 2-[(6-bromo-4-fluoro-benzoimidazol-1-yl)methoxy]ethyl-trimethyl-silane and 2-[(5-bromo-7-fluoro-benzoimidazol-1-yl)methoxy]ethyl-trimethyl-silane was obtained as a waxy beige solid (689 mg, 97% yield). MS m / z 385.2 (MH + ). 1 H NMR (DMSO-d 6)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(d d,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).

[0487] Step 3: Preparation of 2-[7-fluoro-3-(2-trimethylsilylethoxymethyl)benzimidazol-5-yl]oxyethanol: A 3:1 mixture of the above isomers 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 (630 mg, 1.82 mmol) was charged into 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). The resulting mixture was stirred at 120°C for 21 hours. The mixture was cooled to room temperature and then 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 little hexane and then washed once with a saturated aqueous solution of ammonium chloride, twice with water, and once with brine. MgSO 4 The organic layer was dried, filtered and concentrated. The residue was purified by silica gel chromatography using a 30% to 90% EtOAc in hexanes gradient. After combining, concentrating and drying the appropriate fractions, a single isomer of 2-[7-fluoro-3-(2-trimethylsilylethoxymethyl)benzimidazol-5-yl]oxyethanol (307 mg, 52% yield) was obtained as a slowly crystallizing thick colorless oil. MS m / z 327.3 (MH + ). 1 H NMR (DMSO-d 6 )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).

[0488] 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 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 washed with EtOAc and water and a little NaHCO 3 The mixture was diluted. The layers were separated and the aqueous layer was further extracted 4 more times with EtOAc. The combined organic layers were washed with brine and then heated in MgSO 4 The product was added into 4% ethyl acetate, 4% ethyl acetate and 1% ethyl acetate.Then the mixture was dried, filtered and concentrated. The solid residue was purified by silica gel chromatography using a 2% to 20% IPA gradient of EtOAc. The fractions containing the product were combined, concentrated and dried under reduced pressure. 2-[(7-fluoro-3H-benzoimidazole-5-yl)oxy]ethanol (139 mg, 75% yield) was obtained as a white solid. 1 H NMR showed a 85:15 mixture of tautomers. MS m / z 197.2 (MH + ). 1 H NMR (DMSO-d 6 )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).

[0489] Step 5: Prepare the methyl sulfone and methyl sulfoxide mixture (Ar = 2,3-dichlorophenyl) from intermediate A-11 as described above in Example 28 (Step 1), Method B.

[0490] Step 6: Example 100 was prepared by coupling the above 2-[(7-fluoro-3H-benzimidazol-5-yl)oxy]ethanol with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) in DMSO using DIPEA as described in Example 4 in General Procedure A (Step 7).

[0491] Preparation Example 104, Example 112, Example 116 to Example 118:

[0492]

[0493] Step 1: Add 1H-benzimidazole-4-carboxylic acid (1.00 g, 6.17 mmol) in MeOH (6.2 mL) to a 20 mL vial. Then add sulfuric acid (723 μL, 13.6 mmol). Heat the reaction in a 70 °C oil bath for 24 hours. Cool to room temperature. Remove the solvent under reduced pressure. Add the crude material to NaHCO 3 The combined organic layers were washed with water, then with brine, and washed with MgSO 4 Drying, filtration, and concentration under reduced pressure afforded 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 + ).

[0494] Step 2: In N 2 Under atmosphere, a 250 mL round-bottom flask was added with anhydrous THF solution (21 mL) of methyl 1H-benzimidazole-4-carboxylate (750 mg, 4.26 mmol) from the previous step and 2-(trimethylsilyl)ethoxymethyl chloride (982 μL, 5.53 mmol) of stable technology. The mixture was cooled to 0°C, and then a 1 M THF solution of lithium bis(trimethylsilyl)amide (5.5 mL, 5.5 mmol) was slowly added to the reaction over 20 minutes. The reaction was heated to room temperature and stirred overnight. The reaction was cooled to 0°C, and then a 1 M THF solution of lithium aluminum hydride (5.1 mL, 5.1 mmol) was added. The reaction was heated to room temperature and stirred overnight. The reaction was quenched with MeOH at 0°C. It was then adsorbed onto The mixture was added and purified by normal phase silica gel chromatography using a gradient of 0% to 10% IPA 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 (400 MHz, DMSO-d 6 )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).

[0495] Step 3: A 20 mL vial was charged with a MeCN solution (8 mL) of [1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]methanol (200 mg, 0.72 mmol) and DIPEA (500 μL, 2.87 mmol) from the previous step. Methanesulfonyl chloride (167 μL, 2.16 mmol) was then added. The reaction was stirred at room temperature overnight. NaHCO was added. 3 The reaction was quenched with a saturated aqueous solution of MgSO and then extracted three times with EtOAc. The combined organic layers were concentrated in MgSO 4 The resulting mixture was dried over high temperature, filtered, and then concentrated under reduced pressure.

[0496] The crude material was dissolved in DMF (8 mL), and potassium cyanide (187 mg, 2.87 mmol) was added. The reaction was stirred again at room temperature overnight. The reaction was quenched by adding 10% aqueous LiCl solution, and then extracted with EtOAc three times. The combined organic layers were concentrated in MgSO 4 Dry over medium, filter, and then concentrate under reduced pressure. The mixture was then purified by normal phase silica gel chromatography using a 0% to 100% EtOAc in hexanes gradient. 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 + ).

[0497] Step 4: Sodium hydride (60 mg, 1.50 mmol) was added to 4 mL of DMF in a 20 mL vial. 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) in 4 mL of DMF from the previous step was slowly added. After addition, the reaction turned orange. The reaction was warmed to room temperature. After ~30 minutes, the reaction turned a darker orange. The reaction was cooled to 0 °C and then iodomethane (115 μL, 1.85 mmol) was added. After addition, the reaction turned a lighter orange. The reaction was heated to room temperature and stirred overnight. The reaction was quenched with 1 M aqueous HCl and extracted 3 times with EtOAc. The combined organic layers were washed with water, then with brine, and washed with MgSO 4 Dry, filter, and then concentrate under reduced pressure. The mixture was added and purified by normal phase silica gel chromatography using a 0% to 100% EtOAc in hexanes gradient. The desired fractions were collected and concentrated under reduced pressure to give 2-methyl-2-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]propionitrile (134 mg, 69% yield) as an orange-brown oil. MS m / z 316.2 (MH + ).

[0498] Step 5: To a 20 mL scintillation vial was added 2-methyl-2-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]propionitrile (134 mg, 0.426 mmol) from the previous step, followed by DCM (2 mL) and TFA (2 mL). The reaction was stirred at room temperature overnight. The solvent was removed under reduced pressure. The material was dissolved in MeOH and Amberlite IRA-67 resin was added. The resin was filtered off through a cotton plug. The organics were collected and concentrated under reduced pressure to give 2-(1H-benzimidazol-4-yl)-2-methyl-propionitrile; 2,2,2-trifluoroacetic acid (123 mg, 96% yield) as a transparent film. MS m / z 186.0 (MH + ).

[0499] Step 6: Prepare the methyl sulfone and methyl sulfoxide mixture from the appropriate intermediate A-11 as described above in Example 28 (Step 1), Method B.

[0500] Step 7: Example 104, Example 112, Example 116 were prepared by coupling the benzimidazole described in Step 5 with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (with appropriate Ar groups) in DMSO using DIPEA

[0501] To Example 118, as described in Example 4 in General Procedure A (Step 7).

[0502] Preparation Example 105:

[0503]

[0504] Step 1: Preparation of 4-methoxy-1H-imidazo[4,5-c]pyridine: In a pressure 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 at 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 DCM gradient of 0% to 10% MeOH. 4-Methoxy-1H-imidazo[4,5-c]pyridine as a brown solid was obtained (51 mg, 53% yield). MS m / z 150.2 (MH + ).

[0505] Step 2: A mixture of methyl sulfone and methyl sulfoxide was prepared from intermediate A-11 (Ar = 2,3-dichlorophenyl) as described in Method B of Example 28 (Step 1) above.

[0506] Step 3: Example 105 was prepared by coupling 4-methoxy-1H-imidazo[4,5-c]pyridine described in Step 1 with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) in DMSO using cesium carbonate in a similar manner as used in Example 4 (Step 7) of General Method A.

[0507] Preparation of Example 108:

[0508]

[0509] Step 1: Preparation of 3-ethoxy-2-nitroaniline: Sodium ethoxide solution (21% solution in EtOH, 1.63 mL, 4.36 mmol) was added to a solution of 3-fluoro-2-nitroaniline (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, and dried over Na 2 SO 4 and filtered and concentrated. The crude product was purified by column chromatography (silica gel, hexane solution of 0 - 100% EtOAc) to give 3-ethoxy-2-nitroaniline (0.25 g, 94% yield). MS m / z 183.0 (MH + ). 1 H NMR (400 MHz, DMSO-d 6)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).

[0510] 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 iPrOH (4.0 mL) solution of 3-ethoxy-2-nitro-aniline (0.24 g, 1.34 mmol) and formic acid (1.9 mL, 49.6 mmol) from the previous step. The resulting mixture was heated and stirred at 90 ° C for 10 h. The reaction mixture was cooled to room temperature and stirred by Filtered. 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 + ). 1 H NMR (400 MHz, DMSO-d 6 )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).

[0511] Step 3: Prepare the methyl sulfone and methyl sulfoxide mixture from intermediate A-11 (Ar = 2,3-dichlorophenyl) as described above in Example 28 (Step 1), Method B.

[0512] Step 4: Example 108 was prepared by coupling the benzimidazole described in Step 2 with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) using cesium carbonate in DMSO in a similar manner to that used in Example 4 (Step 7) in General Procedure A.

[0513] Preparation Example 110:

[0514]

[0515] Step 1: Preparation of tert-butyl 3-iodoindole-1-carboxylate: Dissolve indole (1.00 g, 8.54 mmol) in 26 mL of DMF in a 200 mL flask. Then add iodine (2.38 g, 9.39 mmol) and potassium hydroxide (1.20 g, 21.3 mmol). Stir the reaction at room temperature for 5 hours. Then add it to 100 mL of Na 2 SO 3 The mixture was then extracted with EtOAc three times. The combined organic layers were washed with water, then brine, and MgSO 4 In 40% ethyl acetate (20% ethyl acetate) 0.25g (2.00mmol) of 4-dimethylaminopyridine (10mg, 0.09mmol) was added to the crude material. The mixture was stirred at room temperature for 18 hours, and then purified by silica gel chromatography using the hexane gradient of 0% to 75% EtOAc. After appropriate fractions were combined and concentrated, tert-butyl 3-iodoindole-1-carboxylate (2.79g, 95% yield) was obtained as brown oil. 1 H NMR (400 MHz, DMSO-d 6 )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).

[0516] 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 a dioxane solution (1.5 mL) of 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). The resulting mixture was heated at 80° C. and stirred for 22 h. The reaction mixture was stirred for 2 hours. The crude product was filtered and purified by column chromatography (silica gel, 0-100% EtOAc in hexanes, then 0-10% methanol in EtOAc gradient) 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 + ).

[0517] Step 3: Prepare the methyl sulfone and methyl sulfoxide mixture from intermediate A-11 (Ar = 2,3-dichlorophenyl) as described above in Example 28 (Step 1), Method B.

[0518] 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 charged into a 4 mL vial along with crude methyl sulfone / methyl sulfoxide (51 mg, 0.09 mmol) from A-11 (Ar = 2,3-dichlorophenyl) and DMSO (1 mL). DIPEA (47 μL, 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 THF solution. 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 in 15 minutes by preparative HPLC using a gradient of 55% to 85% methanol in water (with 0.1% formic acid modifier). 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.

[0519] Preparation Example 111, Example 119 and Example 131:

[0520]

[0521] Step 1 (Example 119: R = Me): Potassium carbonate (0.35 g, 2.56 mmol) and 2-methoxyethanol (0.40 mL, 5.1 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 hours. Water was added and the aqueous mixture was extracted with EtOAc. The organic layers were combined, washed with brine, and washed with NA 2 SO 4 Dry, filter and concentrate. The crude product was purified by column chromatography (silica gel, 0-100% EtOAc in hexanes) to give 3-(2-methoxyethoxy)-2-nitro-aniline (52 mg, 38% yield). MS m / z 213.1 (MH + ).

[0522] Step 2: The substituted benzimidazoles were prepared from the 2-nitroaniline described in Step 1 using the "one pot" procedure described in Method G of Example 113 (Step 2).

[0523] Step 3: As described in Example 28 (Step 1), Method B above, prepare a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 (Ar = 2,3-dichlorophenyl).

[0524] Step 4: Prepare Examples 111, 119, and 131 by coupling the appropriately substituted benzimidazole from Step 2 with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) in DMSO using DIPEA (Example 111) or cesium carbonate (Examples 119 and 131) in a similar manner as used in Example 4 (Step 7) of General Method A.

[0525] Preparation of Example 120:

[0526]

[0527] Step 1: Preparation of 1H-benzo[d]imidazole-4-carbonitrile: Add a DMF solution (4 mL) of 4-bromo-1H-benzo[d]imidazole (99 mg, 0.502 mmol), zinc dicyanide (70.8 mg, 0.603 mmol), and tetrakis(triphenylphosphine)palladium (116 mg, 0.100 mmol) to a 10 mL microwave vial purged with nitrogen three times and equipped with a magnetic stir bar. Heat the reaction to 90 °C and stir for 16 h. Cool the reaction to room temperature, dilute with water, and then extract with EtOAc. Wash the organic layer with NaHCO 3 , brine, dry over Na 2 SO 4 , and then load onto . Purify the crude product by silica gel chromatography using a DCM gradient of MeOH to obtain 1H-benzo[d]imidazole-4-carbonitrile as a light pink solid (31 mg, 43% yield). 1 1H NMR (400 MHz, DMSO-d 6 ) δ: 13.04 (br.s., 1H), 8.45 (s, 1H), 7.90 (d, J = 7.83 Hz, 1H), 7.68 (d, J = 7.83 Hz, 1H), 7.20 - 7.44 (m, 1H). MS m / z 142.2 (MH + ).

[0528] Step 3: As described in Example 28 (Step 1), Method B above, prepare a mixture of methyl sulfone and methyl sulfoxide from intermediate A-11 (Ar = 2,3-dichlorophenyl).

[0529] Step 3: Prepare Examples 111, 119 and 131 by coupling 1H-benzo[d]imidazole-4-carbonitrile in Step 1 with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) in DMSO using DIPEA in a manner similar to that used in Example 4 (Step 7) in General Method A.

[0530] Preparation Example 122:

[0531]

[0532] Step 1: The synthesis of 2-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]acetonitrile is described as the synthesis of Examples 104, 112, 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. A 5 mL DMF solution of 2-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]acetonitrile (240 mg, 0.84 mmol) was slowly added. The reaction was heated to room temperature and then cooled back to 0°C. 1,2-dibromoethane (76 μL, 0.878 mmol) and sodium iodide (138 mg, 0.92 mmol) were added. The reaction was heated to room temperature and then heated to 105°C. The reaction mixture was stirred at this temperature for 24 hours. The reaction was quenched with 1M HCl at room temperature and then extracted 3 times with EtOAc. The combined organic layers were washed with water, then with brine, and washed with MgSO 4 Dry, filter, and then concentrate under reduced pressure. The mixture was then purified by normal phase silica gel chromatography using a 0% to 100% EtOAc in hexanes gradient. The desired fractions were collected and concentrated under reduced pressure to give 1-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]cyclopropanenitrile (111 mg, 42% yield) as a light brown oil. MS m / z 314.2 (MH + ).

[0533] Step 2: In a 20 mL vial, 1-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]cyclopropanenitrile (118 mg, 0.376 mmol) was dissolved in a mixture of DCM (2 mL) and TFA (2 mL). The reaction was stirred at room temperature overnight. The solvent was removed under reduced pressure. The material was dissolved in MeOH and Amberlite IRA-67 resin was added. The resin was filtered off and the filtrate was concentrated under reduced pressure to give 1-(1H-benzimidazol-4-yl)cyclopropanenitrile as a light brown sticky solid; 2,2,2-trifluoroacetic acid (114 mg, 102% yield). MS m / z 184.0 (MH+ ).

[0534] Step 3: Prepare the methyl sulfone and methyl sulfoxide mixture from intermediate A-11 (Ar = 2,3-dichlorophenyl) as described above in Example 28 (Step 1), Method B.

[0535] Step 4: Reaction of 1-(1H-benzimidazol-4-yl)cyclopropanenitrile; 2,2,2-trifluoroacetic acid from step 2 with

[0536] Example 122 was prepared by coupling a crude methyl sulfone / methyl sulfoxide mixture of (Ar = 2,3-dichlorophenyl) in DMSO using DIPEA in a similar manner to that used in Example 4 (step 7) in General Procedure A.

[0537] Preparation Example 123:

[0538]

[0539] Step 1: A 100 mL round-bottom flask equipped with a magnetic stir bar was charged with 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-H 2 O (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. Next, concentrated ammonium hydroxide (2 ml, 29.6 mmol) was added and the reaction was warmed to room temperature and stirred for 24 hours. The reaction was diluted with water and extracted with EtOAc. The organic layer was washed with brine and then with Na 2 SO 4 Dry, filter, and then concentrate under reduced pressure. The product was added and purified by silica gel chromatography using a gradient of 0% to 15% ethanol in ethyl acetate to afford 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 (400 MHz, DMSO-d 6 )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).

[0540] Step 2: Prepare the methyl sulfone and methyl sulfoxide mixture from intermediate A-11 (Ar = 2,3-dichlorophenyl) as described above in Example 28 (Step 1), Method B.

[0541] Step 3: Example 123 was prepared by coupling 1H-benzo[d]imidazole-7-carboxamide in Step 1 with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) in DMSO using DIPEA in a manner similar to that used in Example 4 (Step 7) in General Procedure A.

[0542] Preparation Example 126:

[0543]

[0544] Step 1: Cesium carbonate (8.35 g, 25.6 mmol) was charged into a 50 mL flask and suspended in 6 mL DMSO. Dimethyl malonate (2.7 mL, 30.7 mmol) was then added at room temperature. After stirring the slurry for 10 minutes, 3-fluoro-2-nitro-aniline (800 mg, 5.12 mmol) was added to give a bright orange to red mixture, which was stirred at 70 ° C for 1 hour. 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 3 times with EtOAc, the combined organic layers were washed with brine, and then washed with MgSO 4 Dry, filter and concentrate. The residue is purified by flash chromatography on silica gel using a gradient of 5% to 50% ethyl acetate in hexanes. Dimethyl 2-(3-amino-2-nitrophenyl)malonate (1.225 g, 89% yield) is obtained: MS m / z 269.1 (MH + ). 1 H NMR (DMSO-d 6 )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).

[0545] 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 DMSO and 0.6 mL water. The bright red mixture was heated at 150° C. for 2.5 hours, after which the color darkened somewhat. The mixture was cooled to room temperature and then diluted with water and EtOAc. The two-phase mixture was passed through a small block of The insoluble material was removed by filtration, thereby forming an emulsion, and the layers were separated. The aqueous layer was further extracted 3 times with EtOAc. The combined organic layers were then washed twice with water, once with brine, and then with MgSO 4 Dried, filtered and concentrated. The bright red residue was purified by silica gel flash chromatography using a 15% to 50% EtOAc in hexane gradient. The appropriate fractions were combined, concentrated and dried under reduced pressure. Methyl 2-(3-amino-2-nitro-phenyl) acetate (749 mg, 70% yield) was obtained as a bright orange solid: 1 H NMR (DMSO-d 6 )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).

[0546] Step 3: Methyl 2-(3-amino-2-nitro-phenyl) acetate (545 mg, 2.59 mmol) was charged into a 100 mL flask equipped with a magnetic stirring bar. Then 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 2 drops of acetic acid. The reaction flask was placed under reduced pressure and then hydrogen was introduced. This operation was repeated twice more and the mixture was then vigorously stirred at room temperature under a balloon atmosphere of hydrogen for 24 hours. 0.2 mL of Et 3 N to neutralize AcOH, then pass through a small Filter and rinse with methanol. The filtrate was concentrated to dryness and the residue was purified by flash chromatography on a silica gel column using a 1:1 EtOAc / DCM to 100% EtOAc to 10% IPA EtOAc gradient. The appropriate fractions were combined, 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-d 6)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).

[0547] 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. Then a 1 M THF solution of lithium aluminum hydride (0.25 mL, 0.25 mmol) was added dropwise. Some gas evolution was observed and the clear yellow solution became a milky beige suspension. The mixture was warmed 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 another 10 minutes, the mixture was diluted with 2-3 mL of EtOAc and filtered through The insoluble material was removed by filtration through a plug of 4% paraformaldehyde. The filtrate was concentrated to a residue, which was passed through a silica gel plug using a 30% IPA solution in EtOAc as 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 + ).

[0548] Step 5: Prepare the methyl sulfone and methyl sulfoxide mixture from intermediate A-11 (Ar = 2,3-dichlorophenyl) as described above in Example 28 (Step 1), Method B.

[0549] Step 6: Example 126 was prepared by coupling 2-(1H-benzimidazol-4-yl)ethanol from Step 4 with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) in DMSO using DIPEA in a manner similar to that used in Example 4 (Step 7) in General Procedure A.

[0550] Preparation Example 133:

[0551]

[0552] Step 1: Add 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) to a 100 mL round-bottom flask. Stir the reactants at room temperature for 1 h. After the reaction is complete, add saturated NH 4 The reaction was quenched with Cl and then extracted three times with EtOAc. The organics were collected, washed with brine, separated, and then washed with MgSO 4 Dry, filter, and then concentrate under reduced pressure. The crude material was loaded into The mixture was 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 + ).

[0553] Step 2: In N 2 Under atmosphere, a flame-dried 5 mL microwave vial was charged with a DMF solution (0.30 mL) of 2-[(4-bromobenzimidazol-1-yl)methoxy]ethyl-trimethyl-silane (500 mg, 1.53 mmol) and biboronic acid pinacol ester (776 mg, 3.06 mmol). 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 bubbled with an argon balloon 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 as is in the next step.

[0554] Step 3: To the crude reaction mixture of 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 bubbled with an argon balloon for ~5-10 minutes, sealed, and then heated to 100 °C for 24 hours. After the reaction was complete, the reaction was cooled to room temperature. The reaction was diluted with brine and extracted 3 times with EtOAc. The organic layer was collected and washed with MgSO 4 Dry, filter, and then concentrate under reduced pressure. The crude material was loaded into Then on SiO 2 Purification on the above with MeOH in DCM gave 3-[1-(2-trimethylsilylethoxymethyl)benzimidazol-4-yl]pyridin-4-amine (72 mg, 69% yield) as a brown film. MS m / z 341.2 (MH + ).

[0555] Step 4: A solution of 3-[1-(2-trimethylsilylethoxymethyl)-benzimidazol-4-yl]pyridin-4-amine (72 mg, 0.210 mmol) in DCM (2 mL) and TFA (2 mL) was added to a 20 mL scintillation vial. The reaction was stirred at room temperature for 16 h. After 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 + ).

[0556] Step 5: Prepare the methyl sulfone and methyl sulfoxide mixture from intermediate A-11 (Ar = 2,3-dichlorophenyl) as described above in Example 28 (Step 1), Method B.

[0557] Step 6: Example 133 was prepared by coupling 3-(1H-benzoimidazol-4-yl)pyridin-4-amine TFA salt from Step 4 with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) in DMSO using DIPEA in a manner similar to that used in Example 4 (Step 7) in General Method A.

[0558] Preparation Example 134:

[0559]

[0560] Step 1: Add 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) to a 20 mL scintillation vial. Stir the reaction at room temperature for 2 h. 4 Cl was quenched and then extracted with EtOAc three times. The organic matter was collected and washed with MgSO 4 Dry, filter, and then concentrate under reduced pressure. The crude material was loaded into The resulting mixture was then purified by silica gel chromatography with EtOAc in hexanes to afford 4-bromo-1-[(4-methoxyphenyl)methyl]benzimidazole (735 mg, 91% yield) as a brown oil. 1 H NMR (400 MHz, DMSO-d 6)δ: 8.50 (s, 1H), 7.57 (dd, J = 8.2, 0.8 Hz, 1H), 7.29 (ddd, J = 8.6, 3.1, 2.0 Hz, 2H), 7.14 (t, J = 7.8 Hz, 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, 1.0 Hz, 1H), 7.42 (d, J = 7.8 Hz, 2H), 7.04 (d, J = 9.0 Hz, 2H), 6.90 (m, J = 3.5 Hz, 2H), 5.72 (s, 2H), 3.70 (s, 3H). MS m / z 317.2 (MH + ).

[0561] Step 2: In N 2 Under an atmosphere, a flame-dried 5 mL microwave vial was added with a DMF solution (0.30 mL) of 4-bromo-1-[(4-methoxyphenyl)methyl]benzimidazole (200 mg, 0.63 mmol) and biboronic acid pinacol ester (320 mg, 1.26 mmol) from step 1. 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 bubbled with an argon balloon 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 diluted with brine and then extracted 3 times with EtOAc. The organics were collected and washed with MgSO 4 Dry, through The crude material was loaded into The 4-(4-methoxyphenyl)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzimidazole (82 mg, 36% yield) was obtained by chromatography on silica gel with 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 + ).

[0562] Step 3: Place the mixture in a flame-dried 5 mL microwave vial under N 21-[(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 under atmosphere. 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 bubbled with an argon balloon for ~5-10 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 then extracted 3 times with EtOAc. The organic matter was collected and washed with MgSO 4 Dry, filter, and then concentrate under reduced pressure. The crude material was loaded into The mixture was added to the 4-thiazol-4-yl-benzimidazol-4-yl-pyridin-2-amine solution 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 + ).

[0563] Step 4: A 20 mL scintillation vial was charged with a TFA solution (2 mL) of 3-[1-[(4-methoxyphenyl)methyl]-benzimidazol-4-yl]pyridin-2-amine (15 mg, 0.0445 mmol). The reaction was heated to 80°C and stirred for 96 hours. The solvent was removed under reduced pressure and the product was 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 + ).

[0564] Step 5: Prepare the methyl sulfone and methyl sulfoxide mixture from intermediate A-11 (Ar = 2,3-dichlorophenyl) as described above in Example 28 (Step 1), Method B.

[0565] Step 6: Example 134 was prepared by coupling 3-(1H-benzoimidazol-4-yl)pyridin-2-amine TFA salt from Step 4 with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) in DMSO using DIPEA in a manner similar to that used in Example 4 (Step 7) in General Method A.

[0566] Preparation Example 143:

[0567]

[0568] 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 potassium carbonate (661 mg, 4.78 mmol) was then added, followed by dropwise addition of SEM-Cl (0.42 mL, 2.39 mmol) at room temperature for 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 3 times with EtOAc. The combined organic layers were washed with water, then with brine, and then with MgSO 4 To 4-nitropropene tert-butyl esters (200mg, 51% yield) was added in 4-nitropropene tert-butyl esters (200mg, 51% yield) and 4-nitropropene tert-butyl esters (200mg, 51% yield) in ...

[0569] Step 2: 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 (265 mg, 0.83 mmol) was charged into a 25 mL flask. A solution of diphenyl(vinyl)sulfonium trifluoromethanesulfonate (479 mg, 1.32 mmol) in DMSO (4 mL) was added at room temperature, followed by 1,8-diazabicyclo[5.4.0]undec-7-ene (0.37 mL, 2.48 mmol). The mixture was stirred at this temperature for 17 hours. Then, a solution of 1,8-diazabicyclo[5.4.0]undec-7-ene (0.37 mL, 2.48 mmol) was added. 4 The reaction mixture was diluted with water and EtOAc. The layers were separated and the aqueous layer was extracted twice more with EtOAc. The combined organic layers were washed twice with water and once with brine. MgSO 4The organic layer was dried, filtered and concentrated. The obtained residue was purified by flash chromatography on silica gel using a hexane gradient of 20% to 100% EtOAc, then a 5% IPA solution in EtOAc. The appropriate fractions were combined, concentrated and dried under reduced pressure. An inseparable isomeric mixture of methyl 1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazoles-4-yl)cyclopropane-1-carboxylate and methyl 1-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazoles-7-yl)cyclopropane-1-carboxylate as a pale yellow oil was obtained (250 mg, 87% yield). MS m / z 347.2 (MH + ).

[0570] 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. A 1 M THF solution of lithium aluminum hydride (0.95 mL, 0.95 mmol) was then added dropwise. Some gas evolution was noted. The mixture was warmed to room temperature and stirred for 23 hours. Sodium sulfate decahydrate (0.7 g) was added to the mixture and stirred for another hour. The mixture was then diluted with EtOAc and filtered through 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.

[0571] The first product to elute 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=5.7Hz,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).

[0572] The second product to elute was [1-(1H-benzimidazol-4-yl)cyclopropyl]methanol (47 mg, 31% yield). MS m / z 186.9 (MH - ).

[0573] Step 4: Prepare the methyl sulfone and methyl sulfoxide mixture from intermediate A-11 (Ar = 2,3-dichlorophenyl) as described above in Example 28 (Step 1), Method B.

[0574] Step 5: Example 143 was prepared by coupling [1-(1H-benzimidazol-4-yl)cyclopropyl]methanol from Step 3 with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) in DMSO using DIPEA in a manner similar to that used in Example 4 (Step 7) in General Procedure A.

[0575] Preparation Example 154:

[0576]

[0577] Step 1: Prepare the methyl sulfone and methyl sulfoxide mixture from intermediate A-11 (Ar = 2-methyl-3-fluorophenyl) as described above in Example 28 (Step 1), Method B.

[0578] Step 2: The methyl sulfone / methyl sulfoxide mixture (82 mg, 0.16 mmol) in step 1 was suspended in 1.3 mL 95% EtOH and 0.1 mL water. Then a 1M THF solution of hydrazine (0.77 mL, 0.77 mmol) was added. The resulting mixture was stirred at 45 ° C for 2 hours. The mixture (which had become a light beige suspension) was cooled to room temperature and then diluted with 10 mL of water. The solid in the resulting suspension was collected on hardened filter paper in a Buchner funnel (very slow filtration). The solid was washed with a little 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=8.6H z, 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).

[0579] Step 3: Isoamyl nitrite (0.038 mL, 0.28 mmol) was charged into 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 blowing argon for 5 minutes and then sonicating for another 1 minute. 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 then added in 4 small portions at room temperature (some gas evolution was noted). 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 EDTA aqueous solution. After separation of the layers, the wash was repeated two more times (until no blue color appeared in the aqueous layer). The organic layer was then washed with water and then with brine. MgSO 4 In 40mL 4-bromo-2-nitro-1-pyrimidine-7-yl)amino]-2,4-difluoro-phenyl]-3-fluoro-2-methyl-benzenesulfonamide (80mg, 80% yield) was added in 4-bromo-2-nitro-1-pyrimidine-7-yl)amino-2,4-difluoro-phenyl-3-fluoro-2-methyl-benzenesulfonamide (80mg, 80% yield) to obtain the product.

[0580] 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 charged into 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. The mixture was sonicated and argon was bubbled through the mixture for another 3-4 minutes. The yellow solution was immersed in an oil bath preheated to 110 °C. After stirring at this temperature for 6 hours, the reaction was cooled to room temperature, acidified with a few drops of formic acid, and 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 (2 injections; 50% to 100% MeOH water gradient, 0.1% formic acid). Appropriate fractions were combined and concentrated. The product was lyophilized from a mixture of acetonitrile and water. 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) was obtained as an off-white solid.

[0581] Preparation Example 156:

[0582]

[0583] Step 1: Preparation of Example 156: 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 of Example 113 using the first step Boc protected piperazine. The precursor (63 mg, 0.08 mmol) was dissolved in 0.8 mL 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 a few minutes, the solid was filtered off and the filtrate was concentrated under reduced pressure to a residue and purified by reverse phase preparative HPLC (2 injections, 50% MeOH to 100% MeOH in water 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) as an off-white solid after lyophilization from an acetonitrile / water mixture (Example 156).

[0584] Preparation Example 158:

[0585]

[0586] Step 1: Prepare the starting 4-thiomorpholinobenzimidazole fragment according to Method G. The aqueous solution (10 mL) of (1.42 g, 2.32 mmol) was added to a methanol solution (20 mL) of 4-(1H-benzimidazol-4-yl)thiomorpholine (0.127 g, 0.579 mmol). The resulting mixture was stirred at room temperature for 17 hours. 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 + ).

[0587] Step 2: Prepare the methyl sulfone and methyl sulfoxide mixture from intermediate A-11 (Ar = 2,3-dichlorophenyl) as described above in Example 28 (Step 1), Method B.

[0588] Step 3: Example 158 was prepared by coupling 4-(1H-benzimidazol-4-yl)-1,4-thiazinane 1,1-dioxide from Step 1 with the crude methyl sulfone / methyl sulfoxide mixture from A-11 (Ar = 2,3-dichlorophenyl) in DMSO using cesium carbonate in a manner similar to that used in Example 4 (Step 7) in General Method A.

[0589] Preparation Example 159:

[0590]

[0591] Step 1: Carbomethoxylation of 2,4,5-trifluoroaniline was carried out as described in patent WO2020 / 261156.

[0592] 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) was added 2,3-dichlorobenzenesulfonyl chloride (3.91 g, 15.91 mmol) portionwise at room temperature. The reaction was heated to 70 ° C for 16 hours. The reaction was monitored by LCMS. When the reaction was complete, it was quenched with 1M HCl. The aqueous layer was extracted three times with EtOAc (15 mL). The combined organic layers were washed with brine and washed with MgSO 4 Dried, filtered and evaporated to dryness. The residue was purified by chromatography on a silica gel column using 0-30% EtOAc in hexanes. The 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 + ).

[0593] 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 2M KOH (37 mL, 74.5 mmol) at room temperature. The reaction was stirred overnight at room temperature. When the reaction was complete, it was evaporated to dryness and water (30 mL) and diethyl ether (30 mL) were added to the residue. The aqueous layer was washed twice with ether (20 mL). The aqueous layer was acidified to pH = 2 with 1M HCl. The aqueous layer was extracted three times with EtOAc (30 mL). The combined organic layers were washed with brine and purified by MgSO 4 Drying, filtration and evaporation gave 3-((2,3-dichlorophenyl)sulfonamido)-2,5,6-trifluorobenzoic acid (4.50 g, 91% yield) as a light orange oil. This compound was used as is in the next step. MS m / z 398.0 (MH +).

[0594] Step 4: To a solution of 3-((2,3-dichlorophenyl)sulfonamido)-2,5,6-trifluorobenzoic acid (4.50 g, 11.24 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 to 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 and purified by MgSO 4 Dried, filtered and evaporated to a dark yellow residue. The crude material was purified by chromatography on a silica gel column using 0-50% EtOAc in hexanes. Pure fractions were 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 + ).

[0595] 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 LiOH 4M aqueous solution (17 mL). The pressure vessel was sealed and heated to 100 °C in an oil bath for 1 hour. When the reaction was complete, saturated NH 4 The mixture was quenched with Cl solution. EtOAc was added and the layers were separated. The aqueous layer was extracted twice more with EtOAc (20 mL). The combined organic layers were washed with brine and washed with MgSO 4 Drying, filtration and evaporation gave N-(3-amino-2,4,5-trifluorophenyl)-2,3-dichlorobenzenesulfonamide (1.09 g, 86% yield) as a brown solid. This compound was carried on to the next step without further purification: 1 H NMR (400 MHz, DMSO-d 6 )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.

[0596] Step 6: N-(3-amino-2,4,5-trifluoro-phenyl)-2,3-dichloro-benzenesulfonamide (300 mg, 0.81 mmol) and 7-chloro-2-methylsulfonyl-thiazolo[5,4-d]pyrimidine (A-10, 194 mg, 0.89 mmol) from step 5 were added to ice-cold AcOH (3.2 mL). The reaction was heated at 65 °C overnight and at 85 °C for 2 hours. Another portion of 7-chloro-2-methylsulfonyl-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 extracted with DCM several times. The combined organic layers were washed with brine and washed with MgSO 4 Dry, filter and concentrate under reduced pressure to a brown solid which was used in the next reaction without further purification. MS m / z 552.0 (MH + ).

[0597] Step 7: To a DCM solution (5 mL) of the crude 2,3-dichloro-N-[2,4,5-trifluoro-3-[(2-methylsulfonylthiazolo[5,4-d]pyrimidin-7-yl)amino]phenyl]benzenesulfonamide (238 mg, 0.43 mmol) from step 6 was added 3-chloroperoxybenzoic acid (149 mg, 0.862 mmol). The resulting reaction mixture was stirred at room temperature overnight and then diluted with EtOAc and treated with NaHCO 3 The organic layer was washed with saturated aqueous solution and then with brine. 4 The crude product (mainly methyl sulfone, MS m / z 584.2 (MH + )) was taken to the next step without further purification (66 mg, 26% yield).

[0598] 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 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 passed through a short pad of silica gel and Filter. The filtrate was purified by reverse phase chromatography using 0.1% formic acid as a modifier and a methanol-water gradient. The appropriate fractions were combined and concentrated. The residue was lyophilized from an acetonitrile / water mixture. N-[3-[[2-(Benzimidazole-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) was obtained as a white fluffy powder.

[0599] Biological Activity

[0600] (a) Kinase activity assays for BRAF, CRAF, and ARAF

[0601] Compound preparation: Solid samples of each substance in 1 dram vials were suspended in DMSO (Fisher Scientific) at a stock concentration of 20 mM. The stock solution was stored at -20°C and protected from light. If the solubility of the compound at 20 mM appeared to be problematic, the initial concentration of the DMSO stock solution was changed to 10 mM or 5 mM.

[0602] In vitro enzymatic reactions were used to evaluate the intrinsic activity of compounds against BRAF, CRAF, and ARAF. For BRAF and CRAF, 0.375 nM purified GST-tagged kinase (Cat. No. B4062-10UG and Cat. No. R1656-10UG, respectively, from Millipore Sigma) was mixed with 75 nM kinase-dead MEK1 substrate (Cat. No. 40075; BPS Bioscience) in the presence of 10 μM Ultrapure ATP (Cat. No. V9102; Promega; Part V915A) in the presence and absence of test compounds in 50 mM HEPES pH 7.5, 10 mM MgCl 2 The cells were incubated in a buffer containing 1% EDTA, 0.01% Brij-35, and 2 mM DTT. A separate reaction was performed using MEK1 substrate and ATP as a blank control. The ARAF kinase reaction was strictly identical except that the kinase concentration was raised to 3.75 nM (Catalog No. 1768-0000-1; Reaction Biology).

[0603] For compound treatment, 5 μL / well of the test substance solution was placed in a 384-well replacement plate (Perkin Elmer) and mixed with 2x concentrated kinase reaction. The 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 intrinsic potency), the initial concentration of 100 nM was changed to 1 μM or 0.5 μM and further diluted accordingly. The final concentration of DMSO in the assay was set to 0.05%.

[0604] BRAF and CRAF kinase reactions were performed at 30°C for a total of 2 hours and then stopped by a 1 / 2 dilution in ADP-Glo ​​reagent (Catalog No. V9102; Promega; Part V912C). The reactions were then incubated at room temperature for 1 hour before adding one volume of Kinase Assay Reagent (Catalog No. V9102; Promega; Part V917A). The plates were then equilibrated at room temperature for 30 minutes before luminescence was detected on 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, an internal 100% inhibition control (the average luminescence in the kinase reaction containing only the kinase-dead MEK1 substrate) was subtracted from each data point. The average of the DMSO (vehicle) control (set to 0% inhibition) was determined and used to calculate the % inhibition:

[0605] Inhibition % = 100*(1-((luminescence signal 化合物 ) / (luminous signal DMSO )))

[0606] The ARAF kinase reaction was carried out at 30°C for a total of 2 hours and then terminated by the addition of EDTA to a final concentration of 40 mM. Ultra TM p-MEK 1 / 2 (Ser218 / 222) (PerkinElmer) kit was used to detect the reaction. The reaction was performed using 5 μL of kinase reaction in a 384-well replacement plate (Perkin Elmer) according to the manufacturer's instructions and then incubated at room temperature in a humidified chamber. After the reaction is complete, The signal was recorded on the Synergy Neo2 plate reader (Biotek) of the filter. The effect of each compound dilution on the pMEK signal generated by the ARAF reaction was expressed as % inhibition and calculated as follows. An internal 100% inhibition control (the average luminescence in the kinase reaction containing only the kinase-dead MEK1 substrate) was included in each plate to measure the pMEK background and was subtracted from each data point. The average of the DMSO (vehicle) control (set to 0% inhibition) was also determined and used to calculate the inhibition %:

[0607] Inhibition % = 100*(1-((pMEK signal 化合物 ) / (pMEK signal DMSO )))

[0608] IC 50Values ​​were obtained by plotting kinase inhibition values ​​and fitting dose-activity curves using log(agonist) versus response - variable slope (four parameters) function using GraphPadPrism (V7.0) or Dotmatics Screening Ultra platform. Standards included in the ARAF kinase assay were Bevarafenib (MedChem Express Catalog No. HY-109080; CAS No. 1446113-23-0), LXH254 (MedChem Express Catalog No. HY-112089; CAS No. 1800398-38-2), and BGB283 (Catalog No. HY-18957; CAS No. 1446090-79-4).

[0609] Therefore, all substances reported herein are BRAF, CRAF and ARAF ATP-competitive kinase inhibitors 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, while the ARAF kinase inhibitory potencies of representative analogs are listed in Table A.

[0610] Table A. ARAF kinase inhibition results

[0611]

[0612] For ARAF biochemical kinase assay, * indicates IC 50 >10nM,

[0613] **Indicates IC of 1nM to 10nM 50 Range, *** indicates IC 50 <1nM.

[0614] (b) General cell culture methods

[0615] 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, 5% CO 2The cells were maintained in T175 flasks (Greiner). They were passaged by removing the culture medium, washing once in 10 mL of room temperature phosphate buffered saline (PBS; Wisent) and incubating with 2 mL of 0.05% trypsin (Thermo-Fisher) at 37°C. Trypsin was then inactivated by adding complete growth medium and the cells were then re-plated into T175 culture dishes at appropriate dilutions. All cell lines were regularly tested for mycoplasma contamination. The tissue type and mutation status of each cell line can be seen in Table B.

[0616] Table B. Tumor type and RAS-ERK pathway mutation status of cancer cell lines (CCLs) used for pERK and antiproliferative profiling of the agents described in this application.

[0617] Cell lines Organization Type Mutation Status A375 skin BRAF V600E A101D skin BRAF V600E A2058 skin BRAF V600E RKO colon BRAF V600E HT29 colon BRAF V600E NCIH2087 lung BRAF L597V; KRAS Q61K NCIH1755 lung BRAF G469A NCIH1666 lung BRAF G466V SK-MEL30 skin NRAS-Q61K IPC298 skin NRAS-Q61L HepG2 liver NRAS-Q61L HCT-116 colon KRAS G13D Lovo colon KRAS-G13D SW620 colon KRAS-G12V SW480 colon KRAS G12D NCI-H358 lung KRAS-G12C NCI-H2122 lung KRAS-G12C Calu-6 lung KRAS Q61K Mewo skin NF1 LOF

[0618] (c) pass Ultra TM p-ERK 1 / 2 (Thr202 / Tyr204) measurement in cultured human Phospho-ERK inhibition in cancer cell lines

[0619] Ultra TM p-ERK 1 / 2 (Thr202 / Tyr204) analysis was performed on cells plated at the density specified in Table C in 100 μL complete RPMI-1640 growth medium in 96-well flat-bottom clear culture dishes (Costar). Cells were incubated at 37°C, 5% CO for one hour prior to treatment with a dilution series of compound. 2 The cells were maintained overnight. 2 The cell density in units is equivalent to the number of cells divided by the area of ​​one well of a 96-well plate (0.143 cm 2 ).

[0620] Table C. Number of cells plated per well for each cancer cell line.

[0621]

[0622]

[0623] In the dilution series, 100 μL / well test substance dilutions prepared in complete RPMI-1640 growth medium were added to the cells. The dilution series was selected so that ten concentrations covered the range from 10 μM to 0.33 nM. If necessary, the initial concentration of 10 μM was increased to 100 μM or reduced to 1 μM (as in the case of A375 and H1666 cells, which are generally more sensitive to the compound) and further diluted accordingly. The final concentration of DMSO in the assay was set to 0.5%.

[0624] After treatment, the medium was removed and the cells were lysed in 50 μL 1X AlphaScreen Ultra Lysis Buffer (PerkinElmer). Ultra TM p-ERK 1 / 2 (Thr202 / Tyr204) (PerkinElmer) reactions were performed using 5 μL of cell lysate in a 384-well Proxyplate (PerkinElmer), and the reactions were incubated overnight at room temperature in a humidified chamber. Signals were recorded on an EnVision plate reader (Perkin Elmer).

[0625] The effect of each compound dilution on the pERK signal was expressed as % inhibition and calculated as follows. An internal 100% inhibition control (1 μM trametinib, catalog number HY-10999; MedChem Express; CAS number 871700-17-3) was included in each plate and used as a measure of the pERK background. First, the value obtained for trametinib was subtracted from each data point. The average of the DMSO (vehicle) control (set to 0% inhibition) was determined and used to calculate the inhibition %:

[0626] Inhibition % = 100*(1-((pERK signal 化合物 ) / (pERK signal DMSO )))

[0627] The ability of each compound to inhibit pERK signaling is expressed as IC 50 Values ​​were obtained by plotting the inhibition values ​​for each data point of the dilution series and fitting the obtained curves using a log(agonist) vs. response-variable slope (four parameters) function using GraphPadPrism (V7.0) or Dotmatics Screening Ultra platform.

[0628] When present, paradoxical pERK induction was observed from the pERK IC of the compound. 50 To classify a compound as a paradoxical inducer of pERK, the % inhibition (%Y) of the minimum data point of the dose-activity curve was extrapolated from the negative % inhibition values ​​observed in the curve. MIN ) was set to less than -20%, which is considered to be within the expected assay variation (e.g., %Y MIN = -30% or -50% or -150% are considered to produce paradoxical induction of the pathway, while compounds showing Y MIN = -10% IC 50curve are considered not to produce paradoxical activation of the pathway). Thus, a compound is said to inhibit a pathway in a given cell line without producing paradoxical induction when the following criteria are met:

[0629] 1. The % inhibition at the highest dose tested (30 μM, 10 μM or 1 μM) exceeded 50%.

[0630] 2.IC 50 %Y of the curve MIN Greater than -20%; where Y MIN The IC corresponding to the compound 50 The data point with the lowest value in the curve.

[0631] It is well known to those skilled in the art that some variation in inhibition values ​​is to be expected in such experiments. ±20% Y MIN Values ​​were considered to be within experimental error and not significant. Therefore, only compounds with negative values ​​exceeding the assay variation (approximately >20%) were considered to induce paradoxical activation of signaling cascades and were not included within the scope of the present disclosure. Figure 1 Provided are compounds that induce paradoxical pathway activation (PLX4720, commercially available from Selleck Chemicals; CAS No. 918505-84-7) and IC values ​​for representative compounds as described herein that exhibit unexpected and unique no-induction profiles. 50 Visualization of the curve.

[0632] Figure 1 Shows paradoxical induction of pERK signaling in RAS mutant HCT116 cells (Y MIN >-20%) of compounds as described herein (Example 44 and Example 122) and resulted in a strong induction of this pathway in the same cell line (Y MIN Representative IC of the compound (PLX4720) with 50 Inhibitory dose-response curves.

[0633] Remarkably, according to the above criteria, the compounds of the invention do not induce paradoxical activation of this pathway.

[0634] Example compound 1 to Example compound 159 showed pERK inhibitory activity in the HCT-116 cell line with colon G13D Ras mutation, as shown in Table 3 and Table 4. In addition, some examples also showed inhibition of non-paradoxical induction of pERK signaling in the SW480 colon cell line carrying the G12D allele of KRAS (Table 3 and Table 4). In addition, some examples from Table 3 and Table 4 were also tested in the presence of BRAF V600EInhibition of pERK in A375 cells with a driver mutation and found that they were also active (Table D-1 and Table D-2). All compounds Examples 1 to 159 showed pERK IC in HCT116 cell line 50 Values ​​<30 μM.

[0635] The pERK inhibitory activity of representative compounds as defined herein was also tested on additional tumor cells and showed good to very good pERK inhibitory activity in cancer cell lines carrying various NRAS, KRAS and NF1 alleles and representing a 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 Table D-1 and Table D-2, and for genotypes refer to Table B). The pERK inhibitory activity of the compounds was stronger in cancer cell lines carrying various BRAF alleles (A375, A101D, A2058, RKO, HT29, NCIH2087, NCIH1755 and NCIH1666) (Table D-1 and Table D-2).

[0636] Table D (D-1 and D-2). A panel of cancer cell lines with RAS mutations (genotypes shown in Table B) and BRAF V600E No induction of pERK IC by selected compounds in mutant A375 50 EC 50 value.

[0637] D-1

[0638]

[0639] In pERK: + indicates IC 50 >0.3μM, ++ indicates IC of 0.03μM-0.3μM 50 Range, +++ indicates IC 50 <0.03μM. For proliferation: * indicates proliferating EC 50 >3μM, ** indicates proliferating ECs between 0.3μM and 3μM 50 Range, *** indicates proliferating EC 50 <0.3 μM. N / A: Not Available. Belv.: Bevarafenib. Values ​​in parentheses are % inhibition. Blank indicates that the value was not determined.

[0640] D-2

[0641]

[0642] In pERK: + indicates IC 50>0.3μM, ++ indicates IC of 0.03μM-0.3μM 50 Range, +++ indicates IC 50 <0.03μM. For proliferation: * indicates proliferating EC 50 >3μM, ** indicates proliferating ECs between 0.3μM and 3μM 50 Range, *** indicates proliferating EC 50 <0.3 μM. N / A: Not Available. Belv.: Bevarafenib. Values ​​in parentheses are % inhibition. Blank indicates that the value was not determined.

[0643] For RAS mutant cancer cell lines, pERK IC 50 %Y of the curve min The values ​​were all above -20% and were considered to show minimal or no induction, so the compound did not cause detectable paradoxical activation of the pathway in this panel of cancer cell lines. In contrast, the comparative results for the molecule bevacirafenib (obtained from MedChem Express catalog number HY-109080; CAS number 1446113-23-0) caused mild to strong induction of the pathway in the same cell lines (Y in 10 of the 13 RAS mutant cell lines tested). MIN <-30%).

[0644] (d) Using CellTiter- Reagents for measuring proliferation inhibition of cultured human cancer cell lines (CCLs)

[0645] CellTiter- Viability assays were performed on cells plated at the densities indicated in Table E in 100 μL complete RPMI-1640 growth medium in 96-well flat-bottom white opaque plates (Greiner or Corning) (number of cells plated per well of a 96-well plate for each CCL for CellTiter- Cell viability assay). Measured in cells / cm 2 The cell density in units is equivalent to the number of cells divided by the area of ​​one well of a 96-well plate (0.32 cm 2 The cells were cultured at 37°C and 5% CO 2 The cells were kept overnight at RT and then treated with a dilution series of the compounds for 3 days.

[0646] Table E. Number of cells plated per well of a 96-well plate for CellTiter- Cell viability assay

[0647]

[0648]

[0649] In the dilution series, 100 μL / well test substance dilutions prepared in complete RPMI-1640 growth medium were added to cells initially plated in 100 μL growth medium. The dilution series was selected so that ten concentrations covered the range from 10 μM to 0.33 nM. If necessary (such as in the case of A375 cells, which are more sensitive to the compound), the initial concentration of 10 μM was reduced to 1 μM and further dilutions were performed accordingly. The final concentration of DMSO in the assay was set to 0.5%.

[0650] After 3 days of incubation, the growth medium was removed by aspiration and 60 μL of diluted CellTiter- Reagent (10 μL CellTiter- Reagent + 50 μL PBS). Incubate the cells in CellTiter- The luminescent signal was then acquired on a Synergy Neo2 plate reader (Biotek).

[0651] The effect of each compound dilution on the proliferation of cancer cell lines was expressed as % inhibition and calculated as follows. An internal 100% inhibition control (1 μM trametinib, catalog number HY-10999; MedChem Express; CAS number 871700-17-3) was included in each plate and used as a CellTiter- A measure of signal background. The value obtained for trametinib was subtracted from each data point. The mean of the DMSO (vehicle) control (set to 0% inhibition) was determined and used to calculate the % inhibition:

[0652]

[0653] The ability of each compound to inhibit proliferation was expressed as EC 50 Values ​​were obtained by plotting the effect size for each data point of the dilution series and fitting the obtained curve using a log(agonist) vs. response-variable slope (four parameters) function (using GraphPadPrism (V7.0) or Dotmatics Screening Ultra platform).

[0654] As shown in Tables D-1 and D-2, the active substances showed antiproliferative activity in various NRAS-, KRAS- and NF1-mutant cancer cell lines representing various tissue types (i.e., SK-MEL 30, IPC298, HepG2, HCT-116, Lovo, SW620, SW480, NCI-H358, NCI-H2122, Calu-6 and Mewo; Tables D-1 and D-2 and refer to Table B for genotypes). The antiproliferative activity in cell lines carrying BRAF driver mutations (A375, A101D, A2058, RKO, HT29, NCIH2087, NCIH1755 and NCIH1666) was generally even stronger (Tables D-1 and D-2). Of note, the pERK-reduced IC 50 EC values ​​and antiproliferative activity of substances in KRAS- and BRAF-mutant cell lines 50 The values ​​correlated fairly well with each other (Table D-1 and Table D-2). Therefore, the compounds of the present invention are effective against a variety of tumor types and can be used for these and other indications. This demonstrates the usefulness of the compounds as described herein for treating different types of tumors.

[0655] (e) result

[0656] Exemplary compound structures and biological results are summarized in Tables 3 and 4 below. Each of these tables is followed by its own table summarizing the chemical characteristics and synthesis methods of the compounds.

[0657] Table 3

[0658]

[0659]

[0660]

[0661]

[0662]

[0663]

[0664]

[0665]

[0666] For pERK assay, + indicates IC of 1 μM-10 μM 50 Range, ++ indicates IC of 0.5μM-1μM 50 Range, +++ indicates IC 50<0.5μM. min % value means each IC 50 The lowest value of the curve. min IC values ​​above -20% 50 Compounds below the curve are considered to show minimal or no induction and not cause detectable paradoxical activation of the pathway. For the BRAF biochemical kinase assay, * indicates IC 50 >10nM, ** indicates IC of 1nM-10nM 50 Range, *** indicates IC 50 <1 nM. For CRAF biochemical kinase assay, § indicates IC 50 >50nM, §§ indicates IC of 10nM-50nM 50 Range, §§§ indicates IC 50 <10nM.

[0667] Characterization of the compounds in Table 3

[0668]

[0669]

[0670]

[0671]

[0672]

[0673]

[0674]

[0675]

[0676]

[0677]

[0678]

[0679]

[0680]

[0681]

[0682]

[0683]

[0684]

[0685] Table 4

[0686]

[0687]

[0688] For pERK assay, + indicates IC of 1 μM-10 μM 50 Range, ++ indicates IC of 0.5μM-1μM 50 Range, +++ indicates IC 50 <0.5μM. min % value means each IC 50 The lowest value of the curve. min IC values ​​above -20% 50 Compounds below the curve are considered to show minimal or no induction and not cause detectable paradoxical activation of the pathway. For the BRAF biochemical kinase assay, * indicates IC 50 >10nM, ** indicates IC of 1nM-10nM 50 Range, *** indicates IC 50 <1 nM. For CRAF biochemical kinase assay, § indicates IC 50 >50nM, §§ indicates IC of 10nM-50nM 50 Range, §§§ indicates IC 50 <10nM.

[0689] Table 4 Characterization of Example 159

[0690]

[0691] Various modifications may be made to any of the above-described embodiments without departing from the scope of the present invention.Any reference, patent or scientific literature mentioned in this document is incorporated herein by reference in its entirety for all purposes.

Claims

1. A compound of formula I, or a pharmaceutically acceptable salt or solvate thereof: in: R 1 Selected from substituted or unsubstituted OR 3 , SR 3 NH 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 Selected from substituted C 6 Aryl or C 5-10 Heteroaryl, substituted or unsubstituted C 4-8 Heterocycloalkyl and N(R 3 ) 2 ; R 3 is independently selected at each occurrence from 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 or an electron-withdrawing group; X 2 is selected from H, halogen and electron withdrawing groups; X 3 and X 4 Each is selected from H, halogen, electron withdrawing group, C 1-3 Alkyl, C 3-4 Cycloalkyl and OC 1-3 alkyl.

2. The compound according to claim 1, in, R 2 is substituted C 6 Aryl or C 5-10 Heteroaryl.

3. The compound according to claim 2, in, R 2 For use selected from F, Cl, Br, CN, NO 2 At least one group substituted with C 6 Aryl, and substituted or unsubstituted C 1-3 Alkyl, C 3-4 Cycloalkyl or OC 1-3 alkyl.

4. The compound according to claim 2, in, R 2 A group of the formula: R 4 Select: 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 selected from H, F, Cl, CN, and substituted or unsubstituted C 1-3 Alkyl, C 3-4 Cycloalkyl or OC 1-3 alkyl; R 6 Selected from H, F, Cl, Br, NO 2 , NH 2 , and substituted or unsubstituted C 1-3 Alkyl, C 3-4 Cycloalkyl or OC 1-3 alkyl; R 7 is selected from H, F, Cl, and substituted or unsubstituted C 1-3 alkyl; R 8 is selected from H, F, and substituted or unsubstituted C 1-3 alkyl; Or, R 4 and R 5 , or R 5 and R 6 Together with the carbon atoms to which they are adjacent, they form a substituted or unsubstituted carbocyclic or heterocyclic ring, provided that the heterocyclic ring is not benzoxazolinone; and (---) indicates use as R 2 The bond between the point of attachment and the rest of the molecule; Among them, when R 4 When it is H or F, R 5 , R 6 , R 7 or R 8 At least one of is not H or F; and Among them, when R 5 When it is CN, then R 4 , R 6 , R 7 or R 8 At least one of them is not H.

5. The compound according to claim 4, in, R 4 Selected from H, F, Cl, Br, Me, Et, CN, CHF 2 and CF 3 .

6. The compound according to claim 4 or 5, in, R 5 Selected from H, F, Me, CF 3 , CN and Cl.

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

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

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

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

11. The compound according to claim 4, in: R 4 is selected from Cl and substituted or unsubstituted C 1-3 alkyl; R 5 is selected from H, F, Cl, and substituted or unsubstituted C 1-3 alkyl; R 6 is selected from H, F, Cl, substituted or unsubstituted C 1-3 Alkyl, and substituted or unsubstituted OC 1-3 Alkyl; and R 7 and R 8 Each is H.

12. The compound according to claim 11, in, R 4 Selected from Cl and CH 3 .

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

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

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

16. The compound according to claim 15, in, R 6 CH 3 or OCH 3 .

17. The compound according to claim 1, in, R 2 A group of the formula: in: X 5 Selected from NH, NC 1-3 Alkyl, NC 3-4 Cycloalkyl, O, and S; R 9 , R 10 , R 11 are each independently selected from 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, provided that R 9 and R 11 One of is H and the other is not H; and (---) indicates use as R 2 The bond at the point of attachment between the molecule and the rest of the molecule.

18. The compound according to claim 1, in, R 2 A group of the formula: in: X 5 Selected from NH, NC 1-3 Alkyl, NC 3-4 Cycloalkyl, O, and S; R 9 is 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; R 10 and R 12 are each independently selected from 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 (---) indicates use as R 2 The bond at the point of attachment between the molecule and the rest of the molecule.

19. The compound according to claim 17 or 18, in, 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.

20. The compound according to claim 19, in, R 9 and R 10 are each independently selected from Cl and substituted or unsubstituted C 1-3 alkyl.

21. The compound according to claim 19, in, R 9 and R 10 All are Cl.

22. A compound as claimed in any one of claims 17 to 21, in, X 5 It is O or S, preferably S.

23. The compound according to claim 2, in, R 2 A group of the formula: in: X 9 , X 10 , X 11 , X 12 and X 13 are independently selected from N and C, wherein X 9 , X 10 , X 11 , X 12 and X 13 At least one and at most two of are N; and R 19 , R 20 , R 21 , R 22 and R 23 Selected from H, F, Cl, Br, CN, NO 2 , NH 2 , and substituted or unsubstituted C 1-3 Alkyl, C 3-4 Cycloalkyl or OC 1-3 Alkyl groups, or when they are attached to X 9 , X 10 , X 11 , X 12 or X 13 When N, R 19 , R 20 , R 21 , R 22 and R 23 does not exist; Among them, X 9 and X 13 At least one of is not N; Among them, X 9 and X 13 When one of is N, the other is not N or CH; and (---) indicates use as R 2 The bond at the point of attachment between the molecule and the rest of the molecule.

24. The compound according to claim 1, in, R 2 A group of the formula: in: R 13 is independently selected at each occurrence from 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 both R 13 Together with their adjacent carbon atoms, they form C 3-4 cycloalkyl; and (---) indicates use as R 2 The bond at the point of attachment between the molecule and the rest of the molecule.

25. The compound according to claim 24, in, R 13 For F, Me, OMe, and CH 2 OMe, and n is 1 or 2.

26. The compound according to claim 24 or 25, in, R 13 In 3-position.

27. The compound according to claim 1, in, R 2 N(R 3 ) 2 .

28. The compound according to claim 27, in, R 3 is selected from substituted or unsubstituted C 1-8 Alkyl or C 3-8 Cycloalkyl.

29. The compound according to claim 1, in, R 2 Selected from Group B1 to Group B77.

30. The compound of claim 29, in, R 2 Selected from Group B1 to Group B6.

31. A compound as claimed in any one of claims 1 to 30, in, R 1 OR 3 or SR 3 .

32. The compound of claim 31, in, R 1 For SR 3 .

33. A compound as claimed in any one of claims 1 to 32, in, R 3 is substituted or unsubstituted C 1-8 Alkyl (e.g. C 1-3 alkyl).

34. A compound as claimed in any one of claims 1 to 30, in, R 1 is substituted or unsubstituted C 5-6 Heteroaryl groups.

35. A compound as claimed in any one of claims 1 to 30, in, R 1 is substituted or unsubstituted C 9 Heteroaryl groups.

36. A compound as claimed in any one of claims 1 to 30, in, R 1 is a substituted or unsubstituted group selected from the group consisting of 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 R 1 Attachment to the thiazolopyrimidine nucleus is via the nitrogen atom.

37. A compound as claimed in any one of claims 1 to 30, in, R 1 is substituted or unsubstituted C 4-6 Heterocycloalkyl group.

38. A compound as claimed in any one of claims 1 to 30, in, R 1 is a substituted or unsubstituted group selected from the following: Among them, (---) indicates that it is used as R 1 The bond at the point of attachment between the molecule and the rest of the molecule.

39. The compound of claim 38, in, R 1 is a substituted or unsubstituted group selected from the following: Among them, (---) indicates that it is used as R 1 The bond at the point of attachment between the molecule and the rest of the molecule.

40. A compound as claimed in any one of claims 1 to 39, in, R 1 Substituted by at least one substituent selected from the group consisting of OH, halogen, 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 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N(R 14 ) 2 , CH 2 N(R 16 )C(O)R 15 , CH 2 N(R 16 )SO 2 R 15 , CH 2 N(R 16 )C(O)N(R 14 ) 2 , CH 2 N(R 16 )SO 2 N(R 14 ) 2 and CH 2 N(R 14 ) 2 ; in: R 14 independently selected at each occurrence from 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, they form C 4-10 Heterocycloalkyl groups; R 15 In each occurrence, independently selected from 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 selected at each occurrence from 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 is optionally further substituted.

41. A compound as claimed in any one of claims 1 to 30, in, R 1 A group of the formula: in: R 17 Selected from H, OH, halogen, 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 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N(R 14 ) 2 , CH 2 N(R 16 )C(O)R 15 , CH 2 N(R 16 )SO 2 R 15 , CH 2 N(R 16 )C(O)N(R 14 ) 2 , CH 2 N(R 16 )SO 2 N(R 14 ) 2 and CH 2 N(R 14 ) 2 ; R 27 Selected from H, OH, halogen, 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 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N(R 14 ) 2 , CH 2 N(R 16 )C(O)R 15 , CH 2 N(R 16 )SO 2 R 15 , CH 2 N(R 16 )C(O)N(R 14 ) 2 , CH 2 N(R 16 )SO 2 N(R 14 ) 2 and CH 2 N(R 14 ) 2 , preferably H, halogen, optionally substituted C 1-6 Alkyl, or optionally substituted OC 1-6 alkyl; X 6 is N or CH; and X 7 N and R 18 does not exist; or X 7 C and R 18 Selected from 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 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N(R 14 ) 2 , CH 2 N(R 16 )C(O)R 15 , CH 2 N(R 16 )SO 2 R 15 , CH 2 N(R 16 )C(O)N(R 14 ) 2 , CH 2 N(R 16 )SO 2 N(R 14 ) 2 and CH 2 N(R 14 ) 2 ; Among them, R 14 , R 15 and R 16 As defined in claim 40; wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl or heteroaryl is optionally further substituted; and Among them, (---) indicates that it is used as R 1 The bond at the point of attachment between the molecule and the rest of the molecule.

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

43. The compound of claim 41, in, X 6 For CH.

44. A compound as described in any one of claims 41 to 43, in, X 7 N, R 17 Selected from H, halogen, 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 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N(R 14 ) 2 , CH 2 N(R 16 )C(O)R 15 , CH 2 N(R 16 )SO 2 R 15 , CH 2 N(R 16 )C(O)N(R 14 ) 2 , CH 2 N(R 16 )SO 2 N(R 14 ) 2 and CH 2 N(R 14 ) 2 , and R 18 Absent, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl or heteroaryl is optionally further substituted.

45. The compound of claim 44, in, R 17 Selected from 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 , wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl or heteroaryl is optionally further substituted.

46. ​​The compound of claim 44, in, R 17 Selected from H, F, NH 2 , 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 Heterocycloalkyl.

47. A compound as described in any one of claims 41 to 46, in, R 17 is an optionally substituted C 4-10 Heterocycloalkyl, wherein the heterocycloalkyl is monocyclic or bicyclic and includes 1 to 3 heteroatoms, preferably wherein X 7 is N.

48. The compound of claim 47, in, The heterocycloalkyl group is substituted by at least one selected from F, OH, oxo, CN, C 1-4 Alkyl and OC 1-4 Alkyl group substituted, wherein the C 1-4 The alkyl group is optionally further substituted (e.g., by F, OH, OC 1-3 alkyl, etc. substituted).

49. A compound as described in any one of claims 47 or 48, in, The heterocycloalkyl group is selected from piperidine, piperazine, thiomorpholine, and morpholine groups, or a bicyclic structure (bridged or spiro) containing a piperidine, piperazine, thiomorpholine or morpholine ring.

50. A compound as described in any one of claims 41 to 43, in, X 7 For C.

51. The compound of claim 50, in, R 18 Selected from 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 , CH 2 C(O)R 15 , CH 2 C(O)N(R 14 ) 2 , CH 2 SO 2 R 15 , CH 2 SO 2 N(R 14 ) 2 , CH 2 N(R 16 )C(O)R 15 , CH 2 N(R 16 )SO 2 R 15 , CH 2 N(R 16 )C(O)N(R 14 ) 2 , CH 2 N(R 16 )SO 2 N(R 14 ) 2 and CH 2 N(R 14 ) 2 , wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group is optionally further substituted.

52. The compound of claim 51, in, R 18 Choose your ownC(O)N(R 14 ) 2 , S.O. 2 R 15 Japanese SO 2 N(R 14 ) 2 .

53. A compound as described in any one of claims 50 to 52, in, R 17 Selected from H, OH, halogen, C 1-6 Alkyl, N(R 14 ) 2 , and optionally substituted C 5-10 Heteroaryl.

54. The compound of claim 53, in, R 17 Selected from H, F, NH 2 , and optionally substituted C 5-10 Heteroaryl, preferably H, F or NH 2 .

55. A compound as described in any one of claims 40 to 54, in, R 14 is independently selected at each occurrence from 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, they form an optionally substituted C 4-10 Heterocycloalkyl group.

56. The compound of claim 55, in, Two R's 14 Together with their adjacent nitrogen atoms, they form an optionally substituted C 4-10 Heterocycloalkyl groups, wherein the heterocycloalkyl is monocyclic or bicyclic and includes 1 to 3 heteroatoms.

57. The compound of claim 56, in, The heterocycloalkyl group is substituted by at least one selected from F, OH, oxo, CN, C 1-4 Alkyl and OC 1-4 Alkyl group substituted, wherein the C 1-4 The alkyl group is optionally further substituted (e.g., by F, OH, OC 1-3 alkyl, etc. substituted).

58. A compound as described in any one of claims 55 to 57, in, The heterocycloalkyl group is selected from piperidine, piperazine, thiomorpholine, and morpholine groups, or a bicyclic structure (bridged or spiro) containing a piperidine, piperazine, thiomorpholine or morpholine ring.

59. A compound as described in any one of claims 1 to 30, in, R 1 Selected from: Among them, R 14 , R 17 and R 27 As defined herein, and (---) indicates use as R 1 The bond at the point of attachment between the molecule and the rest of the molecule.

60. The compound of claim 59, in, R 1 Selected from: Among them, R 14 , R 17 and R 27 As defined herein, and (---) indicates use as R 1 The bond at the point of attachment between the molecule and the rest of the molecule.

61. A compound as described in any one of claims 1 to 30, in, R 1 A group of the formula: in: X 15 , X 16 , X 17 and X 18 Independently selected from O, N, S and CR 17 , where R 17 As defined above; Among them, X 15 , X 16 , X 17 and X 18 At most two of are O, N, or S; and Among them, (---) indicates that it is used as R 1 The bond at the point of attachment between the molecule and the rest of the molecule.

62. A compound as described in any one of claims 1 to 30, in, R 1 Selected from Group A1 to Group A550.

63. The compound of claim 62, in, R 1 is selected from the group consisting of 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.

64. The compound of claim 62, in, R 1 is selected from the group consisting of 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.

65. A compound as described in any one of claims 1 to 64, in, X 1 is Cl and X 2 For F.

66. A compound as described in any one of claims 1 to 64, in, X 1 is F and X 2 For H.

67. A compound as described in any one of claims 1 to 64, in, X 1 and X 2 All are F.

68. A compound as described in any one of claims 1 to 67, in, X 3 and X 4 Each is H.

69. A compound as described in any one of claims 1 to 67, in, X 3 is F and X 4 For H.

70. The compound of claim 67, in, The compound has the formula II: Among them, R 1 , R 4 , R 5 and R 6 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 Selected from H, F, Cl, Me, and OMe.

71. The compound of claim 70, in, The compound has the formula IV: Among them, X 6 , X 7 , R 4 , R 5 , R 6 , R 17 , R 27 and R 18 Each is independently as defined above.

72. The compound of claim 70, in, The compound is a compound of formula V: Among them, R 4 , R 5 , R 6 , X 15 , X 16 , X 17 and X 18 Each is independently as defined above.

73. The compound of claim 67, in, The compound has the formula III: Among them, R 1 , R 9 , R 10 , R 12 and X 5 Each is independently as defined above.

74. The compound of claim 73, in, The compound has the formula VI: Among them, R 9 , R 10 , R 12 , R 17 , R 18 , R 27 , X 5 , X 6 and X 7 Each is independently as defined above.

75. The compound of claim 73, in, The compound has the formula VII: Among them, R 9 , R 10 , R 12 , X 5 , X 15 , X 16 , X 17 and X 18 Each is independently as defined above.

76. The compound of claim 1, in, The compound is selected from Examples 1 to 159 as defined herein, or a salt and / or solvate thereof.

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

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

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

80. Use of a compound as defined in any one of claims 1 to 78 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 (RAS pathway disease), or an inflammatory disease or immune system disorder.

81. The use according to claim 80, in, The disease or disorder is selected from the group consisting of tumors and developmental abnormalities.

82. The use according to claim 80 or 81, in, The disease or disorder is associated with a RAF gene mutation (eg, ARAF, BRAF, or CRAF).

83. The use according to any one of claims 80 to 82, in, The disease or disorder is associated with a RAS gene mutation (eg, KRAS).

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

85. The use according to any one of claims 80 to 84, in, The disease or disorder is a tumor.

86. The use according to claim 85, in, The tumor is 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 gland carcinoma, glioma (e.g., ependymoma), lung cancer (e.g., non-small cell lung cancer), head and neck cancer, acute lymphoblastic leukemia, acute myeloid leukemia, non-Hodgkin's lymphoma, and hairy cell leukemia.

87. The use according to claim 85, in, The tumor is selected from colon or colorectal cancer, lung cancer, pancreatic cancer, thyroid cancer, breast cancer, and melanoma.

88. The use according to any one of claims 80 to 87, in, The treatment comprises inhibiting the RAS-ERK signaling pathway without substantially inducing paradoxical pathways.

89. A method for treating a disease or condition selected from the group consisting of a proliferative disease or condition, a developmental abnormality caused by dysregulation of the RAS-ERK signaling cascade (RAS pathway disease), or an inflammatory disease or immune system disorder, the method comprising administering to a subject in need thereof a compound as defined in any one of claims 1 to 78.

90. The method of claim 89, in, The disease or disorder is selected from the group consisting of tumors and developmental abnormalities.

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

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

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

94. The method of any one of claims 89 to 93, in, The disease or disorder is a tumor.

95. The method of claim 94, in, The tumor is 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 gland carcinoma, glioma (e.g., ependymoma), lung cancer (e.g., non-small cell lung cancer), head and neck cancer, acute lymphoblastic leukemia, acute myeloid leukemia, non-Hodgkin's lymphoma, and hairy cell leukemia.

96. The method of claim 94, in, The tumor is selected from colon or colorectal cancer, lung cancer, pancreatic cancer, thyroid cancer, breast cancer, and melanoma.

97. The method of any one of claims 89 to 96, in, The method comprises inhibiting the RAS-ERK signaling pathway without substantially inducing a paradoxical pathway.

98. A method for inhibiting abnormal proliferation of cells, comprising contacting said cells with a compound as defined in any one of claims 1 to 78.

99. The method of claim 98, in, The cell includes a mutated RAF protein kinase (eg, a mutated ARAF, BRAF, or CRAF).

100. The method of claim 98 or 99, in, The cell includes a mutated RAS gene (eg, mutated KRAS).

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

102. The method of any one of claims 98 to 101, in, The cell is selected from the group consisting of a melanoma cell, a thyroid cancer cell (e.g., a papillary thyroid cancer 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 cancer cell), a head and neck cancer cell, an acute lymphoblastic leukemia cell, an acute myeloid leukemia cell, a non-Hodgkin's lymphoma cell, and a hairy cell leukemia cell.

103. The method of any one of claims 98 to 102, in, The cell is selected from the group consisting of a colon or colorectal cancer cell, a lung cancer cell, a pancreatic cancer cell, a thyroid cancer cell, a breast cancer cell, and a melanoma cell.

104. The method of any one of claims 98 to 103, in, The method comprises inhibiting the RAS-ERK signaling pathway without substantially inducing a paradoxical pathway.

105. The method of any one of claims 98 to 104, in, The contacting is performed in vivo.

106. The method of any one of claims 98 to 104, in, The contacting is performed in vitro.

Images