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

Abstract

Compounds, compositions, and their uses in treating proliferative diseases or conditions, e.g., where the proliferative disease or disorder is associated with RAF gene mutations and / or RAS gene mutations. The compounds disclosed have the following formula I or a pharma- ceutical acceptable salt or solvate thereof, wherein R 1 , R 2 , R 3 , X 1 , X 2 , X 3 , X 4 and Y is as defined herein. JPEG2024517505000169.jpg62170

Description

[Technical field]

[0001] Related Applications This application claims priority under applicable law to U.S. Provisional Application No. 63 / 201,222, filed April 19, 2021, the contents of which are incorporated herein by reference in their entirety for all purposes.

[0002] The present disclosure relates generally to pyrimido[5,4-d]pyrimidine compounds, pharmaceutical compositions containing same, and their use in the treatment and prevention of diseases characterized by dysregulation of the RAS-ERK pathway (e.g., cancer, RASopathies). [Background technology]

[0003] 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 growth signals generated by growth factor receptors from the cell membrane to the nucleus. The pathway is dysregulated in the majority of cancers by the activation of receptor tyrosine kinases (RTKs) (e.g., ERBB1, ERBB2, FLT3, RET, KIT) or the activation or inactivation of RAS regulators (SOS1 and NF1), as well as by constitutive activating mutations in the RAS genes (H-, K-, NRAS; 30% of cancers) and the BRAF gene (8% of cancers). The prevalence of KRAS mutations is particularly high in pancreatic cancer (over 90%), colorectal cancer (50%) and lung cancer (30%). On the other hand, BRAF mutations are found particularly frequently in malignant melanoma (70%), thyroid cancer (40%) and colorectal cancer (10%) (mutation frequencies are listed in COSMIC (Catalogue Of Somatic Mutations In Cancer; Wellcome Trust Sanger Institute) release v95, November 24, 2013). th (Based on 2021).

[0004] RAS proteins are small GTPases that transmit extracellular growth signals to intracellular effectors to control important processes such as cell differentiation, proliferation and survival (Nat.Rev.Cancer 2003,3,459). After stimulation of RTKs, physiological activation of RAS is triggered at the cell membrane, which causes GTP to be loaded onto the GTPase and its activation. Activated RAS interacts with and is activated by a series of effector molecules, with RAF kinase being the most important RAS interactor in the context of cancer development (Nature Rev.Drug Discov.2014,13,828). Oncogenic mutations at glycine 12, glycine 13 or glutamine 61 in RAS isoforms cause aberrant and constitutive signaling in human cancers (Nat.Rev.Cancer 2003,3,459) (COSMIC release v95,November 24th 2021).

[0005] Downstream of RAS, mammalian cells express three RAF paralogs (ARAF, BRAF, and CRAF), which share a conserved C-terminal kinase domain (KD) and an N-terminal regulatory region (NTR) that contains a RAS-binding domain (RBD) (Nat.Rev.Mol.Cell Biol.2015,16,281). In unstimulated cells, RAF proteins are sequestered in the cytoplasm as monomers. Binding of GTP-bound activated RAS to the RBD induces membrane anchoring of RAF kinase (Nat.Rev.Mol.Cell Biol.2015,16,281). Concomitantly, RAF proteins undergo side-to-side dimerization of the kinase domain and catalytic activation (Nature 2009,461,542). Activated RAF proteins signal through a phosphorylation cascade from RAF to MEK and then from MEK to ERK, leading to ERK phosphorylation of an array of substrates that trigger cell-specific responses (Nat. Rev. Mol. Cell Biol. 2020, Oct., 21(10), 607).

[0006] Activating mutations in RAF isoforms have so far been mostly restricted to the BRAF gene, but rare mutations have been observed in ARAF and CRAF, highlighting the functional importance of these isoforms (COSMIC release v95, November 24 th The most common cancer mutation in BRAF is a valine to glutamic acid substitution at position 600 (BRAF V600EBRAF is a BRAF-specific activator that enhances BRAF activity by stabilizing its active form (Cell 2004,116,855). Apart from the V600E allele, various mutations occur at other residues (e.g., G466V, D594G, etc.) that cause increased RAF signaling through various mechanisms (Nat. Rev. Mol. Cell Biol. 2015,16,281). These have been classified into three main classes (1-3) depending on the level of dependence on RAS activity and dependence on RAF dimerization (Nature 2017 Aug 10,548(7666),234-238). The critical role of wild-type BRAF and CRAF in mediating RAS-driven oncogenesis by stimulating ERK signaling has been widely validated (Cancer Cell 2011,19,652; Cancer Discov. 2012,2,685; Nat. Commun. 2017,8,15262). Thus, tumor cells rely on elevated and sustained signaling of the RAS-ERK pathway through activation of RAS and RAF, providing strong support for the concept of targeting RAF family kinases in cancer.

[0007] To address existing medical needs, a broad set of ATP-competitive RAF inhibitors have been developed over the past decade (Nat.Rev.Cancer 2017,17,76), primarily targeting 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 target recurrent BRAF V600E It has shown remarkable efficacy against metastatic melanoma with the BRAF allele and is approved to treat this patient population (N. Engl. J. Med. 2011, 364, 2507; Lancet 2012, 380, 358). V600EClinical responses against BRAF-dependent melanomas are due to potent ATP-competitive inhibition of the monomeric form of this specific dimerization-independent BRAF mutant protein (Cancer Cell 2015, 28, 370). Unfortunately, acquired resistance to these agents inevitably develops, often due in part to reactivation of the RAS-ERK pathway through mechanisms that stimulate RAF dimerization. These include upregulation of RTK signaling, RAS mutations, and BRAF V600E This includes amplification or cleavage (Sci. Signal. 2010, 3, ra84; Nature 2010, 468, 973; Nature 2011, 480, 387; Nature Commun. 2012, 3, 724).

[0008] Concurrently, tumors that display RAS activity due to activating RAS mutations or enhanced RTK signaling but are otherwise BRAF wild-type have been shown to be BRAF V600E In contrast, RAF inhibitors have been shown to induce ERK signaling under conditions of elevated RAS activity, thus promoting tumor cell proliferation (Nature 2010,464,431). This counterintuitive phenomenon, known as the paradoxical effect, is also observed in normal tissues that depend on physiological RAS activity and is the basis for some of the side effects seen with RAF inhibitors in melanoma patients, such as the development of new secondary tumors (e.g. squamous cell carcinomas and keratoacanthomas) (Nat.Rev.Cancer 2014,14,455). As a result, BRAF inhibitors have been shown to induce ERK signaling under conditions of elevated RAS activity, thus promoting tumor cell proliferation (Nature 2010,464,431). V600EInhibitors are ineffective and even contraindicated in RAS-driven cancers. The underlying mechanism is due to the ability of the compounds to promote dimerization of RAF kinase domains 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 defined by the binding mode and affinity of the compounds (Nat. Chem. Biol. 2013, 9, 428).

[0009] To circumvent the limitations of first-generation RAF inhibitors in RAS-mutated cancers, two strategies have been recently pursued. The first strategy is based on the observation that paradoxical ERK activation is a dose-dependent phenomenon, i.e., induction occurs at subsaturating inhibitor concentrations, but the pathway is suppressed at saturating concentrations when a compound occupies both protomers of the RAF dimer. Therefore, the first strategy focused on developing molecules with high binding affinity to all RAF paralogs to saturate RAF proteins at lower drug concentrations, thereby suppressing induction of the paradoxical pathway (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 strong RAF dimer induction and thus paradoxically stimulated RAS-ERK signaling, but with a smaller amplitude than that of previous generation RAF inhibitors. Although this class of compounds shows improved properties, they have recently been shown to largely avoid ARAF isoforms, leading to activation of the paradoxical pathway and the emergence of primary and acquired resistance in vitro and in the clinic (Clin Cancer Res. 2021, 27, 2061; Nature 2021, 594, 418). The second strategy consisted in designing compounds that structurally bias the BRAF kinase domain towards an inactive state and thus do not paradoxically induce ERK signaling. This gave rise to the "Paradox Breaker" (PB) molecule PLX8394, an inducer of PLX4032 / vemurafenib (Nature 2015, 526, 583). These molecules inhibit BRAF V600E It has high efficacy against BRAF V600EThis should prove useful for treating RAS-dependent melanoma. However, although PLX8394 does not induce ERK signaling in the RAS-mutant cell lines tested, it remains ineffective and not useful against RAS-mutant tumors. Summary of the Invention [Problem to be solved by the invention]

[0010] There remains a need for inhibitors that potently and consistently inhibit RAS-ERK signaling and cell proliferation in human tumor cells with various RAS and RAF genotypes, and the development of such inhibitors is highly desirable, as it is also important that they do not induce paradoxical pathways in various RAS mutant tumor cell lines. [Means for solving the problem]

[0011] According to one aspect, the present technology relates to compounds of Formula I below, or a pharma- ceutically acceptable salt or solvate thereof: [ka] Where: R 1 is replaced or not replaced OR 3 , S.R. 3 , NH2, NHR 3 , N(R 3 )2, C 3~8 Cycloalkyl, C 4~8 Heterocycloalkyl, C 6~10 Aryl and C 5~10 heteroaryl; R 2 is a substituted C6 aryl and C 5~10 Heteroaryl, substituted or unsubstituted C 4~8 Heterocycloalkyl and N(R 3 )2 is selected; R 3 is independently at each occurrence substituted or unsubstituted; C 1~8Alkyl, C 3~8 Cycloalkyl, C 4~8 Heterocycloalkyl, C 6~10 Aryl and C 5~10 heteroaryl; X 1 is a halogen atom or an electron-withdrawing group; X 2 is selected from H, a halogen atom, and an electron withdrawing group; X 3 and X 4 are H, halogen atoms, electron-withdrawing groups, and C 1~3 Alkyl, C 3~4 Cycloalkyl and OC 1~3 alkyl; Y is H, halogen atom, CN, OH, OC 1~8 Alkyl, NH2, NHC 1~8 Alkyl, N(C 1~8 alkyl)2, and substituted or unsubstituted C 1~8 alkyl; However, the compounds in question are excluded from the following. [ka]

[0012] Compounds of formula I are also defined according to any of the embodiments and examples described throughout this specification.

[0013] According to another aspect, the present technology relates to a pharmaceutical composition for use as defined in any one of the above-described embodiments, wherein the composition comprises a compound as defined herein, together with a pharma- ceutically acceptable carrier, diluent or excipient.

[0014] In a further aspect, the present technology relates to the use of a compound as defined herein for the treatment of a disease or disorder selected from a proliferative disease or disorder, a developmental anomaly caused by dysregulation of the RAS-ERK signaling cascade (RASopathies), or an inflammatory disease or immune system disorder.

[0015] The present technology also relates to a method for treating a disease or disorder selected from a proliferative disease or disorder, developmental anomalies (RASopathies) caused by dysregulation of RAS-ERK signaling cascade, or inflammatory disease or immune system disorder, comprising administering a compound as defined herein to a subject in need of said treatment.A method for inhibiting the abnormal proliferation of cells, comprising contacting said cells with a compound as defined herein, is also contemplated.

[0016] In one embodiment of the above-mentioned use and method, the disease or disorder is selected from neoplasms and developmental anomalies, such as diseases or disorders associated with RAF gene mutations (e.g., ARAF, BRAF or CRAF), diseases or disorders associated with RAS gene mutations (e.g., KRAS), or diseases or disorders associated with both RAF gene mutations and RAS gene mutations. In one embodiment, the disease or disorder is associated with receptor tyrosine kinase mutations or amplifications (e.g., EGFR, HER2), or mutations in downstream regulators of the receptor RAS (e.g., SOS1 gain of function, NF1 loss of function).

[0017] For example, the disease or disorder is a neoplasm, such as a neoplasm selected from melanoma, thyroid cancer (e.g., papillary thyroid cancer), colorectal cancer, ovarian cancer, breast cancer, endometrial cancer, liver cancer, sarcoma, gastric cancer, pancreatic cancer, Barrett's adenocarcinoma, glioma (e.g., ependymoma), lung cancer (e.g., non-small cell lung cancer), head and neck cancer, acute lymphocytic leukemia, acute myeloid leukemia, non-Hodgkin's lymphoma, and hairy-cell leukemia. For example, the neoplasm is selected from colon or colorectal cancer, lung cancer, pancreatic cancer, thyroid cancer, breast cancer, and melanoma. For example, any of the above uses and methods of the present invention include inhibiting the RAS-ERK signaling pathway without substantial induction of the paradoxical pathway.

[0018] Further objects and features of the compounds, compositions, methods and uses of the present invention will become apparent upon reading the non-limiting description of the following illustrative embodiments and examples section, which should not be construed as limiting the scope of the invention. [Brief description of the drawings]

[0019] [Figure 1] FIG. 1 shows representative IC50 inhibition dose-response curves for compounds described herein that do not induce paradoxical induction (Ymin >-20%) of pERK signaling in RAS mutant HCT116 cells (Examples 80 and 81) and a compound (PLX4720; CAS No. 918505-84-7) that causes strong induction (Ymin ~-600%) of the pathway in the same cell line. [Diagram 2] FIG. 2 shows the results of immunoblot analysis of RAS mutant HCT-116 cells treated with a representative compound that does not induce paradoxical induction of pERK or pMEK signaling (Example 80; top panel) and, by comparison, a compound that induces the pathways in the same cell line (PLX4720; bottom panel). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0020] All technical and scientific terms used herein have the same definitions as commonly understood by those skilled in the art to which this technology pertains. Nevertheless, definitions of some terms and expressions used are provided below. To the extent that the definitions of terms in publications, patents, and patent applications incorporated herein by reference are contrary to the definitions set forth herein, the definitions in this specification shall control. The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter disclosed.

[0021] i.Definition

[0022] The chemical structures described herein are drawn according to conventional standards. Also, when an atom, such as a carbon atom, is drawn to have incomplete valences, it is assumed that the valences are satisfied by one or more hydrogen atoms, even if not necessarily explicitly drawn. The hydrogen atoms should be assumed to be part of the compound.

[0023] The terms used herein are for the purpose of describing particular embodiments only and are not intended to be limiting. It should be noted that the singular forms "a," "an," and "the" include the plural forms as well, unless the content clearly dictates otherwise. Thus, for example, a reference to a composition containing "a compound" also refers to a mixture of two or more compounds. It should also be noted that the word "or" is generally used in its sense to include "and / or" unless the context clearly dictates otherwise. Moreover, to the extent that the words "including," "includes," "having," "has," "with," or variations thereof are used in any of the description and / or claims, such words are intended to be inclusive in a manner similar to the word "comprising."

[0024] The term "about" or "approximately" means within an acceptable error range for a particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, "about" can mean within one standard deviation or more than one standard deviation, as per the practice in the art. Alternatively, "about" can mean within a range of 20% or less, preferably 10% or less, more preferably 5% or less, and even more preferably 1% or less of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, preferably within 5-fold, and more preferably within 2-fold, of a value. When a particular value is described in the present specification and claims, unless otherwise indicated, the term "about" should be assumed to mean within an acceptable error range for the particular value.

[0025] As used herein, the terms "compounds", "compounds described herein", "compounds of the present application", "pyrimido[5,4-d]pyrimidine compounds", "pyrimidopyrimidine compounds" and their equivalents refer to compounds described herein, e.g., compounds encompassed by structural formula I, optionally with reference to any of the applicable embodiments, and also include exemplary compounds, e.g., compounds of Examples 1-114, and, where applicable, pharma- ceutically acceptable salts, solvates, esters, and prodrugs thereof. Where zwitterionic forms are possible, the compounds may be depicted as their neutral forms for practical purposes, but it is understood that the compounds also include their zwitterionic forms. The embodiments herein may also exclude one or more of the compounds. The compounds may be identified by either their chemical structure or their chemical name. In the event of a conflict between the chemical structure and the chemical name, the chemical structure shall prevail.

[0026] Unless otherwise stated, structures depicted herein are also meant to encompass all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure, where applicable, such as the R and S configurations for each asymmetric center. Thus, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the compounds of the present application are within the scope of this specification. Unless otherwise stated, therapeutic compounds also encompass all possible tautomers, if any, of an exemplified compound. The term also encompasses isotopically labeled compounds where one or more atoms have an atomic mass different from the atomic mass most abundant in nature. Examples of isotopes that may be incorporated within the compounds of the present invention include: 2 H(D), 3 H(T), 11 C. 13 C. 14 C. 15 N, 18 O. 17 The compounds may also exist in unsolvated forms as well as solvated forms, such as hydrates, as described above. The compounds may also exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated herein and are intended to be within the scope of the present invention.

[0027] When a particular enantiomer is preferred, in some embodiments, it may be provided substantially free of the corresponding enantiomer, or it may be enriched in the enantiomer. "Enantiomerically enriched" means that the compound has a significantly greater proportion of one enantiomer. In some 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 may be isolated from the racemic mixture by any method known to those skilled in the art, such as those described above, including high-pressure liquid chromatography (HPLC) on a chiral support and the formation and crystallization of chiral salts, or may be prepared by asymmetric synthesis.

[0028] The expression "pharmaceutical acceptable salt" refers to a salt of the compound of the present invention that is suitable, within the scope of sound medical judgment, for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic reaction, etc., and commensurate with a reasonable benefit / risk ratio. Such pharmaceutical acceptable salts are well known in the art. For example, SM Berge et al. describe such pharmaceutical acceptable salts in detail in J. Pharmaceutical Sciences, 66:1-19 (1977). The salts can be prepared in situ during the final isolation and purification of the compounds of the present invention, or can be prepared separately by reacting a free base functional group of the compound with a suitable organic or inorganic acid (acid addition salts), or by reacting an acidic functional group of the compound with a suitable organic or inorganic base (base addition salts).

[0029] The term "solvate" refers to a physical association of one of the compounds of the present invention with one or more solvent molecules, such as those listed above, including water and non-aqueous solvent molecules. This physical association may include hydrogen bonding. In some cases, the solvate will be isolable, for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. The term "solvate" encompasses both solution-phase and isolable solvates. Exemplary solvates include, but are not limited to, hydrates, hemihydrates, ethanolates, hemiethanolates, n-propanolates, isopropanolates, 1-butanolates, 2-butanolates, and solvates of other physiologically acceptable solvents, such as Class 3 solvents as described in the International Conference on Harmonization (ICH), Guide for Industry, Q3C Impurities: Residual Solvents (1997). Thus, the compounds described herein also encompass their respective solvates and mixtures thereof.

[0030] As used herein, the expression "pharmaceutical acceptable ester" refers to an ester of a compound formed by a process herein that is hydrolyzable in vivo, including those that decompose readily in the human body to leave the parent compound or a salt thereof. Suitable ester groups include, for example, compounds derived from pharmaceutical acceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic, cycloalkanoic, and alkanedioic acids, where each alkyl or alkenyl moiety advantageously has 6 or fewer carbon atoms. Examples of specific esters include, but are not limited to, formates, acetates, propionates, butyrates, acrylates, ethylsuccinates, and alkyl esters of acidic groups. Other ester groups include sulfonates or sulfates.

[0031] As used herein, the phrase "pharmaceutical acceptable prodrug" refers to a prodrug of a compound formed by a process herein, which prodrug is suitable, within the scope of sound medical judgment, for use in contact with the tissues of humans and lower animals, is not associated with undue toxicity, irritation, allergic response, and the like, and is commensurate with a reasonable benefit / risk ratio and is effective for their intended use. As used herein, "prodrug" means a compound that is convertible in vivo by metabolic means (e.g., hydrolysis) to provide any compound depicted by a formula herein.

[0032] Abbreviations may also be used throughout the present specification, and unless otherwise specified, such abbreviations are intended to have the meanings generally understood in the art. Examples of such abbreviations are Me (methyl), Et (ethyl), Pr (propyl), i-Pr (isopropyl), Bu (butyl), t-Bu (tert-butyl), i-Bu (isobutyl), s-Bu (sec-butyl), c-Bu (cyclobutyl), Ph (phenyl), Bn (benzyl), Bz (benzoyl), CBz or Cbz or Z (carbobenzyloxy), Boc or BOC (tert-butoxycarbonyl), and Su or Suc (succinimide). For greater certainty, additional definitions of certain abbreviations are also included in the introduction to the "Examples" section.

[0033] The number of carbon atoms in a hydrocarbyl substituent is indicated by the prefix "C x ~C y " or "C x~y ", where x is the minimum number of carbon atoms in the substituent and y is the maximum number of carbon atoms in the substituent. However, the prefix "C x ~C y " or "C x~y" is linked to a group that by definition incorporates one or more heteroatoms (e.g., heterocycloalkyl, heteroaryl, etc.), then x and y define the minimum and maximum number of atoms in the ring, respectively, including carbon atoms and the one or more heteroatoms.

[0034] As used herein, the term "alkyl" refers to a saturated, straight or branched chain hydrocarbon moiety that typically contains from 1 to 20 carbon atoms. For example, "C 1~8 Alkyl" contains 1 to 8 carbon atoms. Examples of alkyl moieties include, but are not limited to, methyl moieties, ethyl moieties, propyl moieties, isopropyl moieties, n-butyl moieties, tert-butyl moieties, neopentyl moieties, n-hexyl moieties, heptyl moieties, octyl moieties, and the like.

[0035] As used herein, the term "alkenyl" refers to a straight or branched chain hydrocarbon moiety containing one or more double bonds and typically containing 2 to 20 carbon atoms. For example, "C 2~8 Alkenyl" contains 2 to 8 carbon atoms. Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, l-methyl-2-buten-l-yl, heptenyl, octenyl, and the like.

[0036] As used herein, the term "alkynyl" refers to a straight or branched chain hydrocarbon radical containing one or more triple bonds and typically containing 2 to 20 carbon atoms. For example, "C 2~8 Alkynyl" contains 2 to 8 carbon atoms. Representative alkynyl groups include, but are not limited to, for example, ethynyl, 1-propynyl, 1-butynyl, heptynyl, octynyl, and the like.

[0037] As used herein, the terms "cycloalkyl", "alicyclic", "carbocycle", "carbocyclic" and equivalents refer to monocyclic or polycyclic saturated or partially unsaturated (non-aromatic) carbocyclic rings, such as those described above, including spiro (one atom shared), fused (at least one bond shared) or bridged (two or more bonds shared) carbocyclic ring systems, having 3 to 15 ring members. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopenten-1-yl, cyclopenten-2-yl, cyclopenten-3-yl, cyclohexyl, cyclohexen-1-yl, cyclohexen-2-yl, cyclohexen-3-yl, cycloheptyl, bicyclo[4,3,0]nonanyl, norbornyl, and the like. The term "cycloalkyl" includes both unsubstituted and substituted cycloalkyl groups. For example, the word "C 3~n "Cycloalkyl" refers to cycloalkyl groups having three or more and the indicated "n" carbon atoms in their ring structure. Unless the number of carbons is specifically specified, as used herein, "lower cycloalkyl" groups have at least 3, and up to 8, carbon atoms in their ring structure.

[0038] As used herein, the terms "heterocycle", "heterocycloalkyl", "heterocyclyl", "heterocyclic radical" and "heterocyclic ring" are used interchangeably and refer to a chemically stable 3- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocyclic moiety that is saturated or partially unsaturated and has, in addition to carbon atoms, one or more, preferably one to four, heteroatoms as defined above. When used in reference to a ring atom of a heterocycle, the term "nitrogen atom" includes substituted nitrogen atoms. For example, in a saturated or partially unsaturated ring having one to three heteroatoms selected from an oxygen atom, a sulfur atom or a nitrogen atom, the nitrogen atom may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR (as in N-substituted pyrrolidinyl). A heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a chemically stable structure, and any of the ring atoms can be optionally substituted.Examples of heterocycloalkyl groups are 1,3-dioxolanyl, pyrrolidinyl, pyrrolidonyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydrodithienyl, tetrahydrothienyl, thiomorpholino, thioxanyl, azetidinyl, oxetanyl, thietanyl, homopiperidinyl, oxepanyl, thiamine ... These include, but are not limited to, epanyl, oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, 3-azabicyclo[3,1,0]hexanyl, 3-azabicyclo[4,1,0]heptanyl, quinolidinyl, quinuclidinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinylquinolinyl and the like. Heterocyclyl groups also include groups in which a heterocycle is fused to one or more aryl, heteroaryl, or cycloaliphatic rings, such as indolinyl, 3H-indolyl, chromanyl, chromenyl, phenanthridinyl, 2-azabicyclo[2.2.1]heptanyl, octahydroindolyl, or tetrahydroquinolinyl, where the moiety or point of attachment is on the heterocyclyl ring. Heterocyclyl groups may be monocyclic or bicyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted by heterocyclyl, where the alkyl and heterocyclyl moieties independently may be optionally substituted. For example, the term "C. 3~n "Heterocycloalkyl" refers to a heterocycloalkyl group having 3 or more and indicated "n" atoms in the ring structure, including carbon atoms and heteroatoms.

[0039] 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 include rings that have multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties, as defined herein.

[0040] The term "aryl", whether used alone or as part of a larger moiety such as "aralkyl", "aralkoxy", "aryloxy" or "aryloxyalkyl", refers to an aromatic group having 4n+2 (where n is an integer from 1 to 3) conjugated π (pi) electrons in a monocyclic moiety or in a bicyclic or tricyclic fused ring system having a total of 6 to 15 ring members, where at least one ring in the system is aromatic and each ring in the system contains 3 to 7 ring members. The term "aryl" may be used interchangeably with the term "aryl ring". In certain embodiments herein, "aryl" refers to an aromatic ring system, including but not limited to phenyl, biphenyl, naphthyl, azulenyl, anthracyl, and the like, which may bear one or more substituents. The term "aralkyl" or "arylalkyl" refers to an alkyl residue attached to an aryl ring. Examples of aralkyl include, but are not limited to, benzyl, phenethyl, and the like. Also included within the scope of the term "aryl" as used herein are aromatic rings fused to one or more non-aromatic rings, such as indanyl, indenyl, phthalimidyl, naphthimidyl, fluorenyl, phenanthridinyl, or tetrahydronaphthyl. For example, the term "C 6~n "Aryl" refers to aryl groups having six or more atoms in the ring structure, with the indicated "n" number of atoms.

[0041] The term "heteroaryl", when used alone or as part of a larger moiety such as "heteroaralkyl" or "heteroaralkoxy", refers to an aromatic group having 4n+2 (where n is an integer from 1 to 3) conjugated π (pi) electrons (e.g., having 5 to 18 ring atoms, preferably 5, 6, or 9 ring atoms; having 6, 10, or 14 π electrons shared in 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 atoms, and includes any oxidized form of a nitrogen or sulfur atom, and any quaternized form of a basic nitrogen atom. A heteroaryl may be a single ring or two or more fused rings. As used herein, the term "heteroaryl" also includes groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclic rings, where the portion or point of attachment is on the heteroaromatic ring.Non-limiting examples of heteroaryl groups include thienyl, furanyl (furyl), pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, indolyl, 3H-indolyl, isoindolyl, indolizinyl, benzothienyl (benzothiophenyl), benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzoxazolyl, benzothiazolyl, benzotriazolyl, pyrrolopyridinyl (e.g., pyrrolo[3,2-b] or pyrrolo[3,2-c]pyridinyl), Heteroaryl groups include carbazolyl, acridinyl, phenanthridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Heteroaryl groups may be monocyclic or bicyclic. Heteroaryl groups include rings that are optionally substituted. The term "heteroaralkyl" refers to an alkyl group substituted by a heteroaryl, where the alkyl and heteroaryl portions independently are optionally substituted. Examples include, but are not limited to, pyridinylmethyl, pyrimidinylethyl, and the like. For example, the term "C. 5~n "Heteroaryl" refers to heteroaryl groups having five or more, and the indicated "n" atoms in the ring structure that includes carbon atoms and heteroatoms.

[0042] As described herein, the compounds herein may contain "optionally substituted" moieties. In general, the term "substituted", whether preceded by the term "optionally" or not, means that one or more hydrogen atoms of the specified moiety are replaced with a suitable substituent. Unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents may be the same or different at each position. The combinations of substituents envisioned herein preferably result in the formation of chemically stable or chemically feasible compounds. As used herein, the term "chemically stable" refers to compounds that are substantially unchanged when subjected to conditions that allow for their preparation, detection, and, in some embodiments, their recovery, purification, and use for one or more purposes disclosed herein.

[0043] The term "halo" means a halogen atom, that is, a fluorine, chlorine, bromine or iodine atom, preferably a fluorine or chlorine atom.

[0044] The term "optionally substituted" refers to a group that is substituted or unsubstituted by independently replacing one, two, or more of the hydrogen atoms on the group with a substituent, such as, but not limited to, the following substituents: F, CI, Br, I, OH, COH, alkoxy, oxo, thiooxo, NO, CN, CF, NH, NH alkyl, NH alkenyl, NH alkynyl, NH cycloalkyl, NH aryl, NH heteroaryl, NH heterocycle, dialkylamino, diarylamino, diheteroarylamino, O-aryl, O-aryl- ... alkyl, O-alkenyl, O-alkynyl, O-cycloalkyl, O-aryl, O-heteroaryl, O-haloalkyl, O-heterocycle, C(O)alkyl, C(O)alkenyl, C(O)alkynyl, C(O)cycloalkyl, C(O)aryl, C(O)heteroaryl, C(O)heterocycloalkyl, CO2alkyl, CO2alkenyl, CO2alkynyl, CO2cycloalkyl, CO2aryl, CO2heteroaryl, CO2heterocycloalkyl, OC(O)alkyl, OC(O)alkenyl, OC(O)alkynyl, OC(O)cycloa alkyl, OC(O)aryl, OC(O)heteroaryl, OC(O)heterocycloalkyl, C(O)NH2, C(O)NHalkyl, C(O)NHalkenyl, C(O)NHalkynyl, C(O)NHcycloalkyl, C(O)NHaryl, C(O)NHheteroaryl, C(O)NHheterocycloalkyl, OCO2alkyl, OCO2alkenyl, OCO2alkynyl, OCO2cycloalkyl, OCO2aryl, OCO2heteroaryl, OCO2heterocycloalkyl, OC(O)NH2, OC(O)NHalkyl, OC ... nyl, OC(O)NHalkynyl, OC(O)NHcycloalkyl, OC(O)NHaryl, OC(O)NHheteroaryl, OC(O)NHheterocycloalkyl, NHC(O)alkyl, NHC(O)alkenyl, NHC(O)alkynyl, NHC(O)cycloalkyl, NHC(O)aryl, NHC(O)heteroaryl, NHC(O)heterocycloalkyl, NHCO2alkyl, NHCO2alkenyl, NHCO2alkynyl, NHCO2cycloalkyl, NHCO2aryl, NHCO2heteroaryl, NHCO2heterocycloalkyl,NHC(O)NH2, NHC(O)NH alkyl, NHC(O)NH alkenyl, NHC(O)NH alkynyl, NHC(O)NH cycloalkyl, NHC(O)NH aryl, NHC(O)NH heteroaryl, NHC(O)NH heterocycloalkyl, NHC(S)NH2, NHC(S)NH alkyl, NHC(S)NH alkenyl, NHC(S)NH alkynyl, NHC(S)NH cycloalkyl, NHC(S)NH aryl, NHC(S)NH heteroaryl, NHC(S)NH heterocycloalkyl, NHC(NH)NH2, NHC(NH NH alkyl, NHC(NH)NH alkenyl, NHC(NH)NH alkynyl, NHC(NH)NH cycloalkyl, NHC(NH)NH aryl, NHC(NH)NH heteroaryl, NHC(NH)NH heterocycloalkyl, NHC(NH) alkyl, NHC(NH) alkenyl, NHC(NH) alkynyl, NHC(NH) cycloalkyl, NHC(NH) aryl, NHC(NH) heteroaryl, NHC(NH) heterocycloalkyl, C(NH)NH alkyl, C(NH)NH alkenyl, C(NH)NH alkynyl, C(NH)NH cycloalkyl chloroalkyl, C(NH)NHaryl, C(NH)NHheteroaryl, C(NH)NHheterocycloalkyl, P(O)(alkyl)2, P(O)(alkenyl)2, P(O)(alkynyl)2, P(O)(cycloalkyl)2, P(O)(aryl)2, P(O)(heteroaryl)2, P(O)(heterocycloalkyl)2, P(O)(Oalkyl)2, P(O)(OH)2, P(O)(Oalkenyl)2, P(O)(Oalkynyl)2, P(O)(Ocycloalkyl)2, P(O)(Oaryl)2, P(O)(Oheteroaryl)2, P(O )(Oheterocycloalkyl)2, S(O)alkyl, S(O)alkenyl, S(O)alkynyl, S(O)cycloalkyl, S(O)aryl, S(O)2alkyl, S(O)2alkenyl, S(O)2alkynyl, S(O)2cycloalkyl, S(O)aryl, S(O)heteroaryl, S(O)heterocycloalkyl, SO2NH2, SO2NHalkyl, SO2NHalkenyl, SO2NHalkynyl, SO2NHcycloalkyl, SO2NHaryl, SO2NHheteroaryl, SO2NHheterocycloalkyl, NHSO2alkyl,NHSO2alkenyl, NHSO2alkynyl, NHSO2cycloalkyl, NHSO2aryl, NHSO2heteroaryl, NHSO2heterocycloalkyl, CH2NH2, CH2SO2CH3, alkyl, alkenyl, alkynyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycloalkyl, cycloalkyl, carbocyclic, heterocyclic, polyalkoxyalkyl, polyalkoxy, methoxymethoxy, methoxyethoxy, SH, S-alkyl, S-alkenyl, S-alkynyl, S-cycloalkyl, S-aryl, S-heteroaryl, S-heterocycloalkyl, or methylthiomethyl.

[0045] ii. compound

[0046] The recitation of a list of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of the listed groups. The recitation of embodiments of a variable herein includes those embodiments as any single embodiment or in combination with any other embodiment or portion thereof. The recitation of embodiments herein includes those embodiments. Thus, the following embodiments, where applicable, occur alone or in combination.

[0047] The compounds of the present invention represent a pyrimido[5,4-d]pyrimidine core structure to which defined substituents are attached to achieve the beneficial activity of the product. Exemplary pyrimidopyrimidine compounds as defined herein, or pharma- ceutically acceptable salts or solvates thereof, are represented by the general formula I below: [ka] Where: R 1 is replaced or not replaced OR 3 , S.R. 3 , NH2, NHR 3 , N(R 3 )2, C 3~8 Cycloalkyl, C4~8 Heterocycloalkyl, C 6~10 Aryl and C 5~10 heteroaryl; R 2 is a substituted C6 aryl and 5~10 Heteroaryl, substituted or unsubstituted C 4~8 Heterocycloalkyl and N(R 3 )2 is selected; R 3 independently in each occurrence, substituted or unsubstituted C 1~8 Alkyl, C 3~8 Cycloalkyl, C 4~8 Heterocycloalkyl, C 6~10 Aryl and C 5~10 heteroaryl; X 1 is a halogen atom or an electron-withdrawing group; X 2 is selected from H, a halogen atom, and an electron withdrawing group; X 3 and X 4 are H, halogen atoms, electron-withdrawing groups, and C 1~3 Alkyl, C 3~4 Cycloalkyl and OC 1~3 alkyl; Y is H, halogen atom, CN, OH, OC 1~8 Alkyl, NH2, NHC 1~8 Alkyl, N(C 1~8 alkyl)2, and substituted or unsubstituted C 1~8 alkyl; However, the compounds in question are excluded from the following. [ka]

[0048] For example, the electron withdrawing group is selected from perhaloalkyl (e.g., CF3 or CCl3), CN, NO2, sulfonate, alkylsulfonyl (e.g., SO2Me or SO2CF3), alkylcarbonyl (e.g., C(O)Me), carboxylate, alkoxycarbonyl (e.g., C(O)OMe), and aminocarbonyl (e.g., C(O)NH2). In one embodiment, X 1 is Cl, and X 2 is F, or X 1 is F, and X 2 is H, or X 1 and X 2 are both F. In another embodiment, X 3 and X 4 Each is H. In yet another embodiment, X 3 is F, and X 4 is H.

[0049] According to one example, Y is H and all other groups are as defined herein. According to another example, Y is NH2 and all other groups are as defined herein.

[0050] For example, the aminoarylsulfonamide moiety of formula I is designated L and is selected from the following: [ka] Here, the dashed lines (---) represent bonds.

[0051] In a further embodiment, R 2 is a substituted C6 aryl or C 5~10 Heteroaryl, for example, R 2 is a C6 aryl substituted with at least one group selected from F, Cl, Br, CN, and NO2, and a substituted or unsubstituted C 1~3 Alkyl, C 3~4 Cycloalkyl or OC 1~3 For example, R 2 is a group of the formula: [ka] Where: R 4 is H, F, Cl, Br, CN, and substituted or unsubstituted, C 1~3 Alkyl, C 3~4 Cycloalkyl or OC 1~3 alkyl, for example, R 4 is selected from H, F, Cl, Br, Me, Et, CN, CHF2 and CF3; R 5 is H, F, Cl, CN, and substituted or unsubstituted, C 1~3 Alkyl, C 3~4 Cycloalkyl or OC 1~3 alkyl, for example, R 5 is selected from H, F, Me, CF3, CN and Cl; R 6 is H, F, Cl, Br, NO2, NH2, and substituted or unsubstituted, C 1~3 Alkyl, C 3~4 Cycloalkyl or OC 1~3 alkyl, for example, R 6 is H, F, Cl, B, and substituted or unsubstituted, C 1~3 Alkyl, C 3~4 Cycloalkyl or OC 1~3 alkyl; R 6 is selected from H, F, Cl, Me, Et and OMe; R 7 is H, F, Cl, and substituted or unsubstituted C 1~3 alkyl, for example, R 7 is selected from H, Me, F and Cl; R 8 is H, F, and substituted or unsubstituted C 1~3 alkyl, for example, R 8 is selected from H, Me, and F; Or R 4 and R 5 or R 5and R 6 together with their adjacent carbon atoms form a substituted or unsubstituted carbocyclic or heterocyclic ring, provided that the heterocyclic ring (R 2 ) is not a benzoxazolinone; and (---) represents a bond; Here, R 4 is H or F, then R 5 , R 6 , R 7 or R 8 is other than H or F; and Here, R 5 If is CN, then R 4 , R 6 , R 7 or R 8 At least one of is other than H.

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

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

[0054] In yet another embodiment, R 4 is selected from H, Cl, Br and methyl; R 5 is selected from H, F and Cl; R 6 is selected from H, F, Cl, Me, and OMe; and R 7 and R 8 are each H.

[0055] In a further embodiment, R 4 is Cl, and substituted or unsubstituted C 1~3 alkyl (e.g., Me), preferably R 4 is Cl or Me; R 5 is H, F, Cl, and substituted or unsubstituted C 1~3 alkyl (e.g., Me), preferably R 5 is F, Cl or Me; R 6 is H, F, Cl, substituted or unsubstituted C 1~3 Alkyl (e.g., Me), and substituted or unsubstituted OC 1~3 alkyl (e.g., OCH), preferably R 6 is H or F, or R 6 is Cl, or substituted or unsubstituted C 1~3 Alkyl, or substituted or unsubstituted OC 1~3 alkyl, or CH3 or OCH3; and R 7 and R 8 Each is H. In still other embodiments, R 6 is the replaced C 1~3 It is an alkyl.

[0056] In another example, R 2 is a substituted C5 heteroaryl, for example a group of the formula: [ka] Where: X 5 NH, NC 1~3 Alkyl, NC 3~4 cycloalkyl, O and S; R 9 , R 10 , R 11 are each independently H, F, Cl, CN, and substituted or unsubstituted, C 1~3 Alkyl, C 3~4 Cycloalkyl, C(O)OC 1~3 Alkyl or OC 1~3 alkyl, where R 9 and R 11 is H and the other is not H; and (---) represents a bond.

[0057] Alternatively, R 2 is a radical of the formula: [ka] Where: X 5 NH, NC 1~3 Alkyl, NC 3~4 cycloalkyl, O and S; R 9 is F, Cl, CN, and substituted or unsubstituted, C 1~3 Alkyl, C 3~4 Cycloalkyl, C(O)OC 1~3 Alkyl or OC 1~3 alkyl; R 10 and R 12 each independently represents 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 (---) represents a bond.

[0058] In a preferred embodiment, R 9 and R 10 each independently represents F, Cl, CN, and substituted or unsubstituted, C 1~3 Alkyl, C 3~4 Cycloalkyl, C(O)OC 1~3 Alkyl or OC 1~3 alkyl, preferably Cl, and substituted or unsubstituted C 1~3 alkyl, more preferably R 9 and R 10 are each Cl. In another embodiment, X 5 is O or S, preferably S.

[0059] In another embodiment, R 2 is the replaced C 5~10 Heteroaryl, for example a group of the formula: [ka] Where: X 9 , X 10 , X 11 , X 12 and X 13 is independently selected from N and C, where X 9 , X 10 , X 11 , X 12 and X 13 at least one and not more than two of are N; and R 19 , R 20 , R 21 , R 22 and R 23 is H, F, Cl, Br, CN, NO2, NH2, and substituted or unsubstituted C 1~3 Alkyl, C 3~4 Cycloalkyl or OC 1~3 alkyl or their combined X 9 , X 10 , X 11 , X 12 Or X 13does not exist when N; where X 9 and X 13 is not N; and where X 9 and X 13 When at least one of is N, the other is not N or CH.

[0060] In another example, R 2 is a C5 heterocycloalkyl. For example, R 2 is a radical of the formula: [ka] Where: R 13 independently at each occurrence, F, Cl, and substituted or unsubstituted, C 1~3 Alkyl, C 3~4 Cycloalkyl or C 1~3 alkoxy; n is an integer selected from 0 to 8; or n is 2 to 8, and two R 13 together with their adjacent carbon atoms, C 3~4 forms a cycloalkyl; and (---) represents a bond.

[0061] In one embodiment, R 13 is at position 3. In another embodiment, R 13 is selected from F, Me, OMe, and CHOMe, and n is 1 or 2. For example, R 13 is a methoxy group at the 3-position and n is 1.

[0062] In a further example, R 2 is N(R 3 )2. For example, R 2 is N(R 3 ) 2, and R 3 is substituted or unsubstituted, C 1~8 Alkyl or C 3~8cycloalkyl.

[0063] In yet another embodiment, the compound of formula I is a compound of formula II: or a pharma- ceutically acceptable salt or solvate thereof. [ka] Here, R 1 , R 4 , R 5 and R 6 are each independently as defined above, preferably R 4 is selected from Cl, Br and methyl; R 5 is selected from H, F, Cl and methyl, and R 6 is selected from H, F, Cl, Me and OMe.

[0064] In a further embodiment, the compound of formula I is a compound of formula III below, or a pharma- ceutically acceptable salt or solvate thereof: [ka] Here, R 1 , R 9 , R 10 , R 12 and X 5 are each independently as defined above.

[0065] Example R 2 Groups are selected from groups B1 to B77 defined below. [ka] JPEG2024517505000015.jpg222170JPEG2024517505000016.jpg226170

[0066] Here, (---) represents a bond.

[0067] In one embodiment, R 2is selected from the groups B1 to B37, B41 to B44, B49, B51 to B55, B57, B59, B62 to B67, B71 to B74, B76 and B77, or preferably R 2 is selected from the groups B1 to B33, B36, B41, B42, B51 to B54, B59, B65, B73 and B77, or more preferably, R 2 is selected from the groups B1, B2, B6, B8, B11, B12, B15, B20, B21, B36, B41, B42, B53, B54, B59, B65 and B73, or most preferably selected from the groups B21, B36, B41, B42, B52, B53, B54, B59, B65 and B73.

[0068] In one embodiment, R 1 OR 3 or SR 3 For example, R 1 is SR 3 In various embodiments, R 3 is a substituted or unsubstituted C 1~8 Alkyl (e.g., C 1~3 alkyl).

[0069] In another embodiment, R 1 is a substituted or unsubstituted C aryl group. In another embodiment, R 1 is a substituted or unsubstituted C 4~6 Heterocycloalkyl groups. For example, R 1 is a halogen atom, OH, C 1~6 Alkyl and OC 1~6 C optionally substituted with one or two groups selected from alkyl 4~5 Heterocycloalkyl. For example, R 1 is an N-pyrrolidinyl group substituted with one or two groups selected from F and OH.

[0070] In another embodiment, R 1 is a substituted or unsubstituted C 5~6In a further embodiment, R is a heteroaryl group or a substituted or unsubstituted C heteroaryl group. 1 is a substituted or unsubstituted group selected from thienyl, imidazolyl, pyrazolyl, triazolyl, thiazolyl, pyridyl, pyrimidinyl, indolyl, indazolyl, benzimidazolyl, benzotriazolyl, pyrrolopyridinyl (e.g., pyrrolo[3,2-b]pyridinyl or pyrrolo[3,2-c]pyridinyl), pyrazolopyridinyl (e.g., pyrazolo[1,5-a]pyridinyl), purinyl, imidazopyrazinyl (e.g., imidazo[4,5-b]pyrazinyl), and quinolyl (quinolinyl), preferably R 1 is a substituted or unsubstituted group selected from imidazolyl, pyrazolyl, triazolyl, indolyl, indazolyl, benzimidazolyl, benzotriazolyl, pyrrolopyridinyl (e.g., pyrrolo[3,2-b]pyridinyl or pyrrolo[3,2-c]pyridinyl), pyrazolopyridinyl (e.g., pyrazolo[1,5-a]pyridinyl), purinyl, and imidazopyrazinyl (e.g., imidazo[4,5-b]pyrazinyl), more preferably a substituted or unsubstituted group attached to the pyrimidopyrimidine core through a nitrogen atom.

[0071] R 1 Examples of include groups selected from the following: [ka] where (---) represents a bond, and the group may be further optionally substituted.

[0072] For example, R 1 is a substituted or unsubstituted group selected from: [ka] Here, (---) represents a bond.

[0073] In one embodiment, R 1OH, halogen atoms, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, OC 1~6 Alkyl, C 5~10 Heteroaryl, C 3~10 Cycloalkyl, C 4~10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 )2, SO2R 15 , SO2N(R 14 )2, N(R 16 )C(O)R 15 , N(R 16 )SO2R 15 , N(R 16 )C(O)N(R 14 )2, N(R 16 )SO2N(R 14 )2, N(R 14 )2, P(O)(R 15 )2, CH2C(O)R 15 , CH2C(O)N(R 14 )2, CH2SO2R 15 , CH2SO2N(R 14 )2, CH2N(R 16 )C(O)R 15 , CH2N(R 16 )SO2R 15 , CH2N(R 16 )C(O)N(R 14 )2, CH2N(R 16 )SO2N(R 14 )2 and CH2N(R 14 ) one of the above groups further substituted with at least one substituent selected from Where: R 14 independently in each case, H, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~10 Cycloalkyl, C 4~10 Heterocycloalkyl, C6 aryl, and C 5~10 heteroaryl, or two R 14 together with their adjacent nitrogen atoms, C4~10 Forming a heterocycloalkyl group; R 15 In each case, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~10 Cycloalkyl, C6 aryl, and C 5~10 heteroaryl; and R 16 independently in each case, H, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~10 Cycloalkyl, C6 aryl, and C 5~10 heteroaryl; Here, R 1 (R 14 , R 15 and R 16 The alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl groups contained within the definition of (including within the definition of) may be further optionally substituted.

[0074] In another embodiment, R 1 is a radical of the formula: [ka] Where: R 17 H, OH, halogen atoms, CN, NO2, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, OC 1~6 Alkyl, C 5~10 Heteroaryl, C 3~10 Cycloalkyl, C 4~10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 )2, SO2R 15 , SO2N(R 14 )2, N(R 16 )C(O)R 15 , N(R 16 )SO2R15 , N(R 16 )C(O)N(R 14 )2, N(R 16 )SO2N(R 14 )2, N(R 14 )2, P(O)(R 15 )2, CH2C(O)R 15 , CH2C(O)N(R 14 )2, CH2SO2R 15 , CH2SO2N(R 14 )2, CH2N(R 16 )C(O)R 15 , CH2N(R 16 )SO2R 15 , CH2N(R 16 )C(O)N(R 14 )2, CH2N(R 16 )SO2N(R 14 )2, and CH2N(R 14 )2 is selected; X 6 is N or CH; and X 7 is N, and R 18 is not present; or X 7 is C, and R 18 But, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, OC 1~6 Alkyl, C 5~10 Heteroaryl, C 3~10 Cycloalkyl, C 4~10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 )2, SO2R 15 , SO2N(R 14 )2, N(R 16 )C(O)R 15 , N(R 16 )SO2R 15 , N(R 16 )C(O)N(R 14 )2, N(R 16 )SO2N(R 14 )2, N(R 14 )2, P(O)(R 15 )2, CH2C(O)R15 , CH2C(O)N(R 14 )2, CH2SO2R 15 , CH2SO2N(R 14 )2, CH2N(R 16 )C(O)R 15 , CH2N(R 16 )SO2R 15 , CH2N(R 16 )C(O)N(R 14 )2, CH2N(R 16 )SO2N(R 14 )2, and CH2N(R 14 )2 is selected; Here, R 14 , R 15 and R 16 is as defined above; Here, R 1 (R 14 , R 15 , R 16 , R 17 and R 18 The alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl or heteroaryl included in the definition of (includes) may be further optionally substituted; and Here, (---) represents a bond.

[0075] In another embodiment, R 1 is the basis of the formula: [ka] Where: X 15 , X 16 , X 17 , and X 18 are independently O, N, S and CR 17 where R 17 is as defined above; where X 15 , X 16 , X 17 and X 18 Not more than two of these are O, N, or S.

[0076] In one embodiment, the compound of formula I is a compound of formula IV or V below, or a pharma- ceutically acceptable salt or solvate thereof. [ka] [ka] Here, R 4 , R 5 , R 6 , R 17 , R 18 , X 6 , X 7 , X 15 , X 16 , X 17 and X 18 are each independently as defined herein, preferably R 4 is selected from Cl, Br and methyl; R 5 is selected from H, F, Cl and methyl; R 6 is selected from H, Cl, F, Me and OMe.

[0077] In further embodiments, the compound of formula I is a compound of formula VI or formula VII: or a pharma- ceutically acceptable salt or solvate thereof. [ka] [ka] Here, R 9 , R 10 , R 12 , R 17 , R 18 , X 5 , X 6 , X 7 , X 15 , X 16 , X 17 and X 18 are each independently as defined herein.

[0078] In one embodiment of the above formula, X 6 is N. In another embodiment, X 6 is CH.

[0079] In another embodiment, X 7 is N and R 17 are H, OH, CN, and C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, OC 1~6 Alkyl, C 5~10 Heteroaryl, C 3~10 Cycloalkyl, C 4~10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 )2, SO2R 15 , SO2N(R 14 )2, N(R 16 )C(O)R 15 , N(R 16 )SO2R 15 , N(R 16 )C(O)N(R 14 )2, N(R 16 )SO2N(R 14 )2, N(R 14 )2, P(O)(R 15 )2, CH2C(O)R 15 , CH2C(O)N(R 14 )2, CH2SO2R 15 , CH2SO2N(R 14 )2, CH2N(R 16 )C(O)R 15 , CH2N(R 16 )SO2R 15 , CH2N(R 16 )C(O)N(R 14 )2, CH2N(R 16 )SO2N(R 14 )2 and CH2N(R 14 ) 2, and R 18 does not exist, where R 14 , R 15 , R 16 or R 17The alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl or heteroaryl in the formula (I) may be optionally substituted, and preferably R 17 is C 1~6 Alkyl, C 5~10 Heteroaryl, C 4~10 Heterocycloalkyl, N(R 14 )2, N(R 16 )C(O)R 15 , N(R 16 )SO2R 15 , C(O)N(R 14 )2 and SO2N(R 14 )2, where R 14 , R 15 , R 16 or R 17 In the above, the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl or heteroaryl may be further optionally substituted. For example, R 17 is H, NH2 and optionally substituted, C 5~10 Heteroaryl or C 4~10 heterocycloalkyl, preferably R 17 is an optionally substituted C 5~10 Heteroaryl or C 4~10 It is a heterocycloalkyl.

[0080] In a further embodiment, R 17 is optionally substituted, C 4~10 Heterocycloalkyl, where the heterocycloalkyl may be monocyclic or bicyclic and contains 1 to 3 heteroatoms, preferably, where X 7 is N. In a preferred embodiment, the heterocycloalkyl is, for example, F, OH, oxo, CN, C 1~4 Alkyl and OC 1~4 alkyl, wherein the C 1~4 Alkyl is (e.g., F, OH, OC 1~3For example, the heterocycloalkyl may be selected from optionally substituted piperidine, piperazine, thiomorpholine, and morpholine groups, or bicyclic structures (bridged or spiro) containing a piperidine, piperazine, thiomorpholine, or morpholine ring.

[0081] In a further embodiment, X 7 is C, for example X 7 is C and R 18 is C 1~6 Alkyl, C 5~10 Heteroaryl, C 3~10 Cycloalkyl, C 4~10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 )2, SO2R 15 , SO2N(R 14 )2, N(R 16 )C(O)R 15 , N(R 16 )SO2R 15 , N(R 16 )C(O)N(R 14 )2, N(R 16 )SO2N(R 14 )2, N(R 14 )2, P(O)(R 15 )2, CH2C(O)R 15 , CH2C(O)N(R 14 )2, CH2SO2R 15 , CH2SO2N(R 14 )2, CH2N(R 16 )C(O)R 15 , CH2N(R 16 )SO2R 15 , CH2N(R 16 )C(O)N(R 14 )2, CH2N(R 16 )SO2N(R 14 )2 and CH2N(R 14 )2, where R 14 , R 15 , R 16 or R 18The alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl or heteroaryl in the formula (I) may be optionally further substituted, and preferably R 18 is C(O)N(R 14 )2, SO2R 15 and SO2N(R 14 In a subclass of these embodiments, R 17 is H, OH, C 1~6 Alkyl, N(R 14 ) 2, and optionally substituted C 5~10 For example, R 17 is H, NH2 and optionally substituted, C 5~10 heteroaryl, preferably H or NH2.

[0082] In yet another embodiment, R 14 is independently at each occurrence H, optionally substituted C 1~6 Alkyl, optionally substituted C 3~10 Cycloalkyl, optionally substituted C 4~10 Heterocycloalkyl and optionally substituted C 5~6 heteroaryl, or two R 14 together with their adjacent nitrogen atoms, optionally substituted C 4~10 It forms a heterocycloalkyl group.

[0083] In another embodiment, R 17 is N(R 14 )2, where R 14 together with their adjacent nitrogen atoms, C 4~10 Form a heterocycloalkyl group, where the heterocycloalkyl may be monocyclic or bicyclic and contains 1 to 3 heteroatoms, preferably where X 7 is N. In preferred embodiments, the heterocycloalkyl is, for example, F, OH, oxo, CN, C 1~4 Alkyl and OC1~4 alkyl, wherein the C 1~4 Alkyl is (e.g., F, OH, OC 1~3 For example, the heterocycloalkyl is selected from optionally substituted piperidine, piperazine, thiomorpholine, and morpholine groups, or bicyclic structures (bridged or spiro) containing a piperidine, piperazine, thiomorpholine, or morpholine ring.

[0084] In another example, R 1 is selected from the following: [ka] Here, R 14 is as defined herein, and (---) represents a bond.

[0085] In a further example, R 1 is selected from the following: [ka] Here, R 14 is as defined herein, and (---) represents a bond.

[0086] Each substituent R 1 group (C group), R 2 Further subembodiments including the groups (B), Y and L are also presented in the Examples section. Exemplary combinations are also set forth in Tables 2 to 5 below. Representative preferred compounds of Examples 1 to 163 are also described herein.

[0087] More specifically, the exemplary R 1 The groups are selected from C1 to C493 groups defined as follows: [ka] JPEG2024517505000028.jpg198170JPEG2024517505000029.jpg206170JPEG20245175050000 30.jpg225170JPEG2024517505000031.jpg200170JPEG2024517505000032.jpg185170JPEG202 4517505000033.jpg221170JPEG2024517505000034.jpg217170JPEG2024517505000035.jpg2 25170JPEG2024517505000036.jpg201170JPEG2024517505000037.jpg199170JPEG2024517505 000038.jpg204170JPEG2024517505000039.jpg205170JPEG2024517505000040.jpg220170JP EG2024517505000041.jpg223170JPEG2024517505000042.jpg203170JPEG2024517505000043. jpg229170JPEG2024517505000044.jpg207170JPEG2024517505000045.jpg225170JPEG202451 7505000046.jpg226170JPEG2024517505000047.jpg217170JPEG2024517505000048.jpg84170

[0088] Here, (---) represents a bond.

[0089] In one embodiment, R 1 is selected from the group consisting of C1 to C493, or R 1 is selected from the groups C1 to C23, C27, C60, C69, C71 to C73, C81 to C83, C88, C114, C182 to C184, C196, C220, C223 to C226, C275, C292, C310, C312, C313, C323, C346, C376, C402, C404, C414, C418, C419, C434, C435, C438, C440, C441, C472, C483, C488 and C490, for example, R 1is selected from the groups C1, C3, C5, C7, C22, C23, C27, C60, C69, C73, C81 to C83, C88, C182 to C184, C196, C224 to C226, C313, C323, C376, C402, C404, C414, C418, C419, C438 and C488, for example, selected from the groups C7, C22, C23 and C60, or selected from the groups C183, C323, C376, C414, C418, C419, C438 and C488.

[0090] The following embodiments are illustrative of R 1 (C1~C493) group, R 2 Combinations of (B1 to B77) groups and L (L1 to L4) groups are shown below: C1-L-B1;C1-L-B2;C1-L-B3;C1-L-B4~70;C1-L-B71;C1-L-B72;C1-L-B73;C1-L-74;C1-L-B75~B77; C2-L-B1;C2-L-B2;C2-L-B3;C2-L-B4~70;C2-L-B71;C2-L-B72;C2-L-B73;C2-L-B74;C2-L-B75~B77; C3-L-B1;C3-L-B2;C3-L-B3;C3-L-B4~70;C3-L-B71;C3-L-B72;C3-L-B73;C3-L-B74;C3-L-B75~B77; C4~C488-L-B1;C4~C488-L-B2;C4~C488-L-B3;C4~C488-L-B4~70;C4~C488-L -B71;C4~C488-L-B72;C4~C488-L-B73;C4~C488-L-B74;C4~C488-L-B75~B77; C489-L-B1;C489-L-B2;C489-L-B3;C489-L-B4~B70;C489-L-B71;C489-L-B72;C489-L-B73;C489-L-B74;C489-L-B75~B77; C490-L-B1;C490-L-B2;C490-L-B3;C490-L-B4~B70;C490-L-B71;C490-L-B72;C490-L-B73;C490-L-B74;C490-L-B75~B77; C491~C493-L-B1;C491~C493-L-B2;C491~C493-L-B3;C491~C493-L-B4~B70; C491~C493-L-B71;C491~C493-L-B72;C491~C493-L-B73;C491~C493-L-B74;or C491~C493-L-B75~B77.

[0091] In one embodiment, the compound is as defined in Formula I, wherein: R 1 is selected from the group consisting of C1 to C493, or R 1 is selected from the groups C1 to C23, C27, C60, C69, C71 to C73, C81 to C83, C88, C114, C182 to C184, C196, C220, C223 to C226, C275, C292, C310, C312, C313, C323, C346, C376, C402, C404, C414, C418, C419, C434, C435, C438, C440, C441, C472, C483, C488 and C490, for example, R 1 is selected from the groups C1, C3, C5, C7, C22, C23, C27, C60, C69, C73, C81 to C83, C88, C182 to C184, C196, C224 to C226, C313, C323, C376, C402, C404, C414, C418, C419, C438 and C488; R 2 is selected from the groups B1, B2, B8, B11, B12, B20 to B23, B34 to B37, B41 to B44, B49, B51 to B54, B57, B59, B62 to B67, B71 to B74 and B77; and L is a group selected from L1 to L4, and Y is H or NH2, preferably Y is H.

[0092] In another embodiment, the compound is as defined in formula I, wherein R 1 is selected from C7, C22, C23, C60, C73, C81, C83, C183, C376, C404, C414, C418, C419, C438 and C488; R 2 is selected from B12, B21, B36, B41, B42, B52 to B54, B59, B65 and B73, L is a group selected from L1 to L4, and Y is H or NH2, preferably Y is H.

[0093] Exemplary compounds as defined herein include each of the single compounds listed below in Tables 2, 3, 4, and 5, from Example 1 through Example 163.

[0094] Examples of preferred compounds include, namely, Examples 31, 36, 40, 51, 55-60, 69, 72, 80-83, 88, 93, 94, 96-122, 124-147, 149, 151-160, 162 and 163 from Tables 3, 4 and 5. Examples of more preferred compounds include Examples 80-83, 93, 94, 96, 98-101, 104, 106, 111, 112, 114-116, 119, 120, 122, 125, 128-134, 139, 142, 144-146, 153, 155, 157, 159 and 162 from Tables 3 and 5.

[0095] It is understood that any of the above compounds may be in any amorphous, crystalline or polymorphic form, including any salt or solvate form, or a combination thereof. The compounds herein may be further modified by adding various functional groups via any synthetic means defined herein to enhance selective biological properties. Such modifications are known in the art and include those that increase biological penetration into a given biological system (e.g., blood, lymphatic system, central nervous system), those that increase oral availability, those that increase solubility to allow administration by injection, those that change metabolism, and those that change excretion rate.

[0096] These compounds can be prepared by conventional chemical synthesis, such as those described above and illustrated in the schemes and examples of this disclosure. As can be appreciated by those of skill in the art, additional methods of synthesizing compounds of the formulae herein will be apparent to those of skill in the art. In addition, various synthetic steps can be performed in an alternative sequence or order to provide the desired compounds.

[0097] iii. Methods, Uses, Formulations and Administration

[0098] As used herein, the term "effective amount" refers to an amount of a drug or pharmaceutical agent that elicits the biological or medical response of a tissue, system, animal or human that is being sought, for example, by a researcher or clinician. Moreover, the term "therapeutically effective amount" refers to an amount that results in the treatment, cure, prevention, amelioration of a disease, disorder or symptoms thereof, or a reduction in the rate of progression of a disease or disorder, compared to a corresponding subject who has not been administered such amount. This term also includes within its scope an amount effective to enhance normal physiological function.

[0099] As used herein, the terms "treatment," "treat," and "treating," as described herein, refer to reversing a disease or disorder or one or more symptoms thereof, alleviating one or more symptoms, delaying the onset of one or more symptoms, or inhibiting the progression of one or more symptoms. In some embodiments, treatment may be administered after one or more symptoms have developed. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and / or in light of genetic or other susceptibility factors). Treatment may also be continued after symptoms have disappeared, e.g., to prevent or delay their recurrence.

[0100] In one embodiment, the disease or condition to be treated is a proliferative disease or disorder, or a kinase-mediated disease or disorder. More specifically, the disease or disorder to be treated includes a proliferative disease or disorder, developmental disorders caused by dysregulation of the RAS-ERK signaling cascade (RASopathies), an inflammatory disease, or an immune system disorder.

[0101] According to some examples, the proliferative disease or disorder to be treated is a neoplasm, inflammatory disease or condition, or developmental abnormality with constitutive activating mutations in the RAS and / or RAF genes (e.g., KRAS and / or ARAF, BRAF, or CRAF mutations). The disease or disorder may also be further associated with receptor tyrosine kinase mutations or amplifications (e.g., EGFR, HER2), or mutations in RAS downstream regulators of the receptor (e.g., SOS1 gain of function, NF1 loss of function). For example, the compounds defined herein are useful for treating RAF mutations (e.g., BRAF V600E The compounds of the present invention are inhibitors of signaling enzymes (e.g., BRAF and CRAF) that are involved in controlling cell proliferation not only in tumors carrying BRAF-associated BRAF kinase (BRK) but also, importantly, in the context of mutated RAS-driven cancers. Thus, the compounds of the present invention may be used, for example, for the treatment of diseases that are linked to the activity of these signaling enzymes and are characterized by excessive or abnormal cell proliferation.

[0102] According to one embodiment, the disease or disorder is characterized by uncontrolled cell proliferation, i.e., a "proliferative disorder" or "proliferative disease." More specifically, these diseases and disorders relate to abnormal conditions characterized by cells with autonomous proliferation capacity, i.e., rapidly proliferating cell proliferation that typically forms distinct masses that show partial or total lack of structural organization and functional coordination with normal tissues.

[0103] For example, proliferative disorders or diseases are defined as "neoplasms," "neoplastic disorders," "neoplasia," "cancer," and "tumors," which terms are collectively meant to include hematopoietic neoplasms (e.g., lymphomas or leukemias) and solid neoplasms (e.g., sarcomas or carcinomas), which include, for example, precancerous and cancerous growths, or cancerous processes, metastatic tissues, or malignantly transformed cells, tissues, or organs of any kind, regardless of histopathological type or stage of invasion. Hematopoietic neoplasms are malignant tumors affecting hematopoietic structures (structures associated with the formation of blood cells) and components of the immune system, and include, for example, leukemias (associated with leukocytes (white blood cells) and their precursors in the blood and bone marrow) and lymphomas (associated with lymphocytes) arising from the myeloid, lymphoid, or erythroid lineages. Solid neoplasms include sarcomas, which are malignant neoplasms that arise from connective tissues, such as muscle, cartilage, blood vessels, fibrous tissue, fat, or bone. Solid neoplasms also include carcinomas, which include malignant neoplasms arising from epithelial structures, external epithelium (e.g., skin, and the lining of the digestive tract, lungs, and cervix), and internal epithelium covering various glands (e.g., breast, pancreas, thyroid). Examples of neoplasms include leukemia, and hepatocellular carcinoma, sarcoma, vascular endothelial carcinoma, breast cancer, central nervous system cancer (e.g., astrocytoma, gliosarcoma, neuroblastoma, oligodendroglioma, and glioblastoma), prostate cancer, lung and bronchial cancer, laryngeal cancer, esophageal cancer, colon cancer, colorectal cancer, gastrointestinal cancer, melanoma, ovarian and endometrial cancer, renal and bladder cancer, liver cancer, endocrine cancer (e.g., thyroid), and pancreatic cancer. For example, the disease or disorder is selected from colon cancer, lung cancer, pancreatic cancer, thyroid cancer, breast cancer, and skin cancer. Examples of neoplasms include melanoma, papillary thyroid carcinoma, colorectal cancer, ovarian cancer, breast cancer, endometrial cancer, liver cancer, sarcoma, gastric cancer, Barrett's adenocarcinoma, glioma (including ependymoma), lung cancer (including non-small cell lung carcinoma), head and neck cancer, acute lymphocytic leukemia, acute myeloid leukemia, non-Hodgkin's lymphoma, and hairy cell leukemia.

[0104] In one embodiment, a patient presenting with one of the above-mentioned hematopoietic or solid neoplasms has previously been treated with an inhibitor targeting the RAS-ERK pathway (including an RTK, RAF, MEK or ERK inhibitor) and has acquired resistance to the inhibitor, including standard of care, such as vemurafenib, dabrafenib, cobimetinib, trametinib, YERVOY, OPDIVO, or a combination of these agents.

[0105] In one embodiment, the disease to be treated is defined by developmental abnormalities caused by dysregulation of the RAS-ERK signaling cascade (RASopathies: e.g., Noonan syndrome, Costello syndrome, LEOPARD syndrome, cardiac anomaly syndromes, and hypertrophic cardiomyopathy).

[0106] In one embodiment, the disease to be treated is defined as an inflammatory disease or immune system disorder, examples of which include inflammatory bowel disease, Crohn's disease, ulcerative colitis, systemic lupus erythematosis (SLE), rheumatoid arthritis, multiple sclerosis, thyroiditis, type 1 diabetes, sarcoidosis, psoriasis, allergic rhinitis, asthma, and chronic obstructive pulmonary disease (COPD).

[0107] In one embodiment, the compounds defined herein inhibit RAS-ERK signaling and cell proliferation in tumor cells harboring at least one mutated RAS or RAF genotype without inducing or substantially inducing the paradoxical pathway.

[0108] As used herein, the term "patient or subject" refers to an animal, e.g., a mammal. Thus, a subject can refer to, for example, mice, rats, dogs, cats, horses, cows, pigs, guinea pigs, primates, such as those listed above, including humans, and the like. Preferably, the subject is a human.

[0109] Thus, the present specification further relates to a method of treating a subject, e.g. a human subject, e.g. a subject suffering from a proliferative disease or disorder, e.g. a subject suffering from a RAF mutated cancer and / or a mutated RAS driven cancer, comprising administering to a subject in need of such treatment a therapeutically effective amount of a compound as defined herein.

[0110] In some embodiments, the present disclosure provides a method for treating a disorder (described herein) in a subject, comprising administering a compound of the present disclosure to a subject identified as needing such treatment. Identifying a patient who needs treatment for the above-mentioned disorder is within the ability and knowledge of a person skilled in the art. Certain methods for identifying patients at risk of developing the above-mentioned disorder that can be treated by the subject method are understood in the medical arts, such as family history and the presence of risk factors associated with the development of the disease state in the subject patient. A medically skilled person can easily identify such candidate patients, for example, using clinical tests, physical examination, medical / family history, and genetic determination.

[0111] A method for evaluating the effectiveness of a treatment in a subject includes determining pretreatment symptoms of the disorder by methods well known in the art, and then administering a therapeutically effective amount of a compound of the present invention to the subject. After an appropriate period (e.g., 1 week, 2 weeks, 1 month, 6 months) after administration of the compound, the symptoms of the disorder are determined again. Modulation (e.g., reduction) of symptoms and / or biomarkers of the disorder (e.g., pERK or pMEK) indicates the effectiveness of the treatment. The symptoms and / or biomarkers of the disorder can be determined periodically throughout the treatment period. For example, the symptoms and / or biomarkers of the disorder can be checked every few days, weeks, or months to evaluate the further effectiveness of the treatment. A reduction in the symptoms and / or biomarkers of the disorder indicates that the treatment is effective.

[0112] In some embodiments, a therapeutically effective amount of a compound defined herein can be administered to a patient alone or in a composition in admixture with a pharma- ceutically acceptable carrier, adjuvant, or vehicle.

[0113] The expression "a pharma- ceutically acceptable carrier, adjuvant, or vehicle" and equivalent expressions refer to a non-toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound in which it is formulated. Pharmaceutically acceptable carriers, adjuvants, or vehicles that may be used in the compositions of the present disclosure include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and wool fat.

[0114] The compositions described herein may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, bucally, or via an implanted reservoir. As used herein, the term "parenteral" includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intracavity, and intracranial injection or infusion techniques. Other methods of administration also include intradermal or transdermal administration.

[0115] Liquid dosage forms for oral administration include, but are not limited to, pharma- ceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compounds, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing and emulsifying agents, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, and mixtures thereof. In addition to inert diluents, oral compositions may also contain adjuvants, such as wetting agents, emulsifying and suspending agents, surfactants, sweeteners, flavorings, and aromatics.

[0116] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions, can be formulated according to known techniques using suitable dispersing or wetting agents and suspending agents. Sterile injectable preparations can also be sterile injectable solutions, suspensions or emulsions in non-toxic parenterally acceptable diluents or solvents, for example, solutions in 1,3-butanediol. Among the acceptable vehicles and solvents that can be used are water, Ringer's solution, USP and isotonic sodium chloride solution. In addition, sterile fixed oils are commonly used as solvents or suspending media. For this purpose, any brand of fixed oil can be used, for example, the above-mentioned fixed oils, including synthetic mono- or diglycerides. In addition, fatty acids, such as oleic acid, are used in the preparation of injectables.

[0117] 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 which can be dissolved or dispersed in sterile water, or other sterile injectable medium prior to use.

[0118] In order to prolong the effect of a given compound, it is often desirable to slow the absorption of the compound by subcutaneous or intramuscular injection. This can be accomplished by using a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the compound then depends on its rate of dissolution, which in turn depends on the size of the crystals and the crystalline form. Alternatively, delayed absorption of parenterally administered compounds is accomplished by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are prepared by forming microencapsule matrices of the compound in biodegradable polymers, such as polylactide-polyglycolide. Depending on the ratio of the compound to the polymer and the nature of the polymer used, the release rate of the compound can be controlled.

[0119] Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes and microemulsions which are compatible with body tissue.

[0120] Compositions for rectal administration are preferably suppositories, which can be prepared by mixing the compounds herein with a suitable non-irritating excipient or carrier, such as cocoa butter, polyethylene glycol or a suppository wax, which is solid at ambient temperature but liquid at body temperature and therefore will melt in the rectum and release the active compound.

[0121] 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, pharma- ceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate, and / or a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and silicic acid, b) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone (PVP), sucrose and acacia, c) humectants, such as glycerol, d) disintegrants, such as glycerol, e) glycerol, f) glycerol, g) glycerol, h) glycerol, h) glycerol, i) glycerol, j) glycerol, j) glycerol, sucrose ... For example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate, e) solution retarders such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as cetyl alcohol and glyceryl monostearate, h) adsorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also contain buffering agents.

[0122] Solid compositions of a similar type can also be used as fillers for soft and hard filled gelatin capsules, using additives such as lactose or milk sugar and high molecular weight polyethylene glycols. The solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical formulation art. They can optionally contain opacifying agents, and also can be of a composition that releases one or more active ingredients only, or preferentially, in a certain part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type can also be used as fillers for soft and hard filled gelatin capsules, using additives such as lactose or milk sugar and high molecular weight polyethylene glycols, etc.

[0123] The composition may also be in microencapsulated form with one or more additives 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, release controlling coatings and other coatings well known in the pharmaceutical formulation art. In such solid dosage forms, the active compound may be admixed with at least one inert diluent, such as sucrose, lactose or starch. Such dosage forms may also contain, as is common practice, additional substances other than inert diluents, such as tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may also contain buffering agents. They may optionally contain opacifying agents, and they may also be of a composition that releases one or more active ingredients only or preferentially in a certain part of the intestinal tract, optionally with a delay. Examples of embedding compositions that can be used include polymeric substances and waxes.

[0124] Dosage forms for topical or transdermal administration of the compounds herein include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active ingredient is mixed under sterile conditions with a pharma- ceutical acceptable carrier and any preservatives or buffers that may be required, as appropriate. Ophthalmic formulations, ear drops, and eye drops are also contemplated within the scope of the present invention. In addition, the present invention contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of the compound into the body. Such dosage forms can be prepared by dissolving or dispensing the compound in a suitable medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0125] The pharma- ceutically acceptable compositions provided herein may also be administered by nasal aerosol or inhalation.Such compositions are prepared according to techniques well known in the art of pharmaceutical formulation and may be prepared as a solution in saline using benzyl alcohol or other suitable preservatives, absorption enhancers to enhance bioavailability, fluorocarbons, and / or other conventional solubilizing or dispersing agents.

[0126] The pharma- ceutically acceptable compositions provided herein can be formulated for oral administration. Such formulations can be administered with or without food. In some embodiments, the pharma- ceutically acceptable compositions of the present disclosure are administered without food. In other embodiments, the pharma- ceutically acceptable compositions of the present disclosure are administered with food.

[0127] The amount of compound that can be combined with a carrier material to produce a composition in a single dosage form will vary depending upon the patient to be treated and the particular mode of administration. The provided compositions can be formulated so that a dosage of 0.01-100 mg / kg body weight / day of the inhibitor can be administered to a patient receiving these compositions.

[0128] It should also be understood that the specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, the judgment of the treating physician, and the severity of symptoms associated with the proliferative disease or disorder. The amount of compound provided in the composition will also depend on the particular compound in the composition.

[0129] The compounds or compositions described herein may be administered using any amount and any route of administration effective for treating or reducing the severity of symptoms as contemplated herein. The exact amount required will vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the infection, the specific drug, its mode of administration, and the like. The compounds provided are preferably formulated into unit dosage forms for ease of administration and uniformity of dosage. As used herein, the expression "unit dosage form" refers to a physically separate unit of drug appropriate for the patient to be treated. However, it will be understood that the total daily usage of the compounds and compositions of the present disclosure will be determined by the attending physician within the scope of sound medical judgment.

[0130] The pharma- ceutically acceptable compositions of the present disclosure can be administered to humans and other animals orally, rectally, parenterally, intraperitoneally, topically (by powder, ointment, or drops), buccally, as an oral or nasal spray, etc., depending on the severity of the infection being treated. In certain embodiments, provided compounds can be administered orally or parenterally at a dosage of about 0.01 mg / kg to about 50 mg / kg, preferably about 1 mg / kg to about 25 mg / kg, of the subject's body weight per day, one or more times per day to obtain the desired therapeutic effect.

[0131] It will be understood that the total daily usage of the compounds and compositions herein can be determined by the attending physician within the scope of sound medical judgment. The total daily inhibitory dose of the compounds herein administered to a subject in single or divided doses can be, for example, in an amount of 0.01 to 50 mg / kg body weight, more usually 0.1 to 25 mg / kg body weight. Single dose compositions can contain such amounts or multiples thereof to make up a daily dose. In one embodiment, a therapeutic regimen according to the present invention comprises administering to a patient in need of such treatment, in a single or multiple doses, from about 10 mg to about 1000 mg of one or more compounds herein per day.

[0132] Depending on the disease or disorder to be treated, additional therapeutic agents may also be present in the compositions of the present disclosure or may be administered separately as part of the dosing regimen, e.g., an additional chemotherapeutic agent. Non-limiting examples of additional therapeutic agents that can be used in combination with the compounds of the invention include anti-proliferative compounds, such as aromatase inhibitors; anti-estrogens; anti-androgens; gonadorelin agonists; topoisomerase I inhibitors; topoisomerase II inhibitors; microtubule activators; alkylating agents; retinoids, carotenoids, tocopherols; cyclooxygenase inhibitors; MMP inhibitors; antimetabolites; platin compounds; methionine aminopeptidase inhibitors; bisphosphonates; anti-proliferative antibodies; heparanase inhibitors; inhibitors of Ras oncogenic isoforms; telomerase inhibitors; proteasome inhibitors; compounds used in the treatment of hematological malignancies; kinesin spindle protein inhibitors; Hsp90 inhibitors; mTOR inhibitors; PI3K inhibitors; Flt-3 inhibitors; CDK4 / 6 inhibitors; HER2 inhalants (Herceptin, Trastuzumab; EGFR inhibitors (Iressa, Tarceva, Nerlynx, Tykerb, Erbitux); RAS inhibitors; MEK inhibitors (Trametinib, Binimetinib, Cobimetinib); ERK inhibitors (Ulixertinib); anti-PD-1 antibodies (Opdivo), (Keytruda); anti-CTLA4 antibodies (Yervoy); anti-tumor antibiotics; nitrosoureas; compounds which target / increase protein or lipid kinase activity or compounds which target / increase protein or lipid phosphatase activity or further anti-angiogenic compounds.

[0133] The treatment may also be supplemented with other treatments or interventions, such as surgery, radiation therapy (e.g., gamma irradiation, neutron irradiation, electron beam radiation, proton beam therapy, brachytherapy, and systemic radioisotopes), biologic response modifiers (e.g., interferons, interleukins, tumor necrosis factor (TNF)), and drugs used to reduce side effects.

[0134] The description of an embodiment of a variation herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof. The description of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof.

[0135] Working Example

[0136] List of abbreviations: Ac: Acetyl AcOEt: ethyl acetate AcOH: acetic acid Ar: Aryl ATCC: American Type Culture Collection ATP: Adenosine triphosphate BINOL: [1,1'-binaphthalene]-2,2'-diol Boc: tert-butyloxycarbonyl BOP: (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate br: Broad BSA: bovine serum albumin CCL: cancer cell lines DCE: 1,2-dichloroethane DCM: dichloromethane DIEA (or DIPEA): N,N-diisopropylethylamine (Huenig's base) DME: 1,2-dimethoxyethane DMF: N,N-dimethylformamide DMSO: Dimethyl sulfoxide DTT: Dithiothreitol EA: Ethyl acetate EC 50 : Half-maximal effective concentration ECL: enhanced chemiluminescence EDTA: Ethylenediaminetetraacetic acid Et2O: Diethyl ether EtOH: Ethanol Eu: Europium FBS: Fetal bovine serum GST: glutathione S-transferase HATU: O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate HEPES: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid Het: Heterocycle Hex: Hexane HRMS: high resolution mass spectrometry HPLC: High performance liquid chromatography HRP: Horseradish peroxidase I C 50 :half-maximal inhibitory concentration IPA or iPrOH: Isopropanol LCMS: liquid chromatography mass spectrometry MeCN: Acetonitrile MS: mass spectrometry NMP: N-methylpyrrolidone NMR: nuclear magnetic resonance ON: Overnight PBS: Phosphate-buffered saline pERK: phosphorylated extracellular signal-regulated kinase PMB: para-methoxybenzyl PMSF: Phenylmethylsulfonyl fluoride Rf: retention factor RPMI-1640: Roswell Park Memorial Institute medium RT: room temperature SDS: Sodium dodecyl sulfate SDS-PAGE: sodium dodecyl sulfate-polyacrylamide gel electrophoresis SEM: Trimethylsilylethoxymethyl SNAr: Nucleophilic aromatic substitution TBST: 0.2% Tween (登録商標) Tris-buffered saline containing -20 TBTU: O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium trifluoroborate TEV: tobacco etch virus protease TFA: Trifluoroacetic acid THF: tetrahydrofuran TLC: Silica gel thin layer chromatography TR-FRET: Time-resolved fluorescence resonance energy transfer Ts: para-toluenesulfonate Y 最小 :minimal data point of a dosage-activity curve

[0137] The following non-limiting examples are illustrative embodiments and should not be construed as further limiting the scope of the present invention. These examples will be better understood with reference to the accompanying figures.

[0138] The examples presented herein below provide synthesis and experimental results obtained for certain exemplary compounds. As is well known to those skilled in the art, reactions are carried out in an inert atmosphere (nitrogen or argon) as necessary to protect reaction components from air and moisture. Temperatures are given in degrees Celsius (°C). Solution percentages and ratios express volume to volume relationships unless otherwise noted. Reactants used in the examples below may be obtained as described herein or, if not described herein, are themselves commercially available or may be prepared from commercially available materials by methods known in the art. Flash chromatography is performed on silica (SiO2) at 254 nm using a Teledyne Isco Rf Combiflash instrument using commercially available normal phase silica. Mass spectrometry is recorded using an electrospray mass spectrometer. NMR was recorded on a 400 MHz Varian instrument.

[0139] Preparative HPLC was performed using Agilent equipment with a Phenomenex-Kinetex C18, (21x100mm, 5μm) column at a flow rate of 20mL / min (RT) and UV detection at 220nm and 254nm. The mobile phase consisted of solvent A (5% MeOH, 95% water + 0.1% formic acid) and solvent B (95% MeOH, 5% water + 0.1% formic acid) unless otherwise noted. When specified herein, 0.05% TFA or 0.1% AcOH or other additives were sometimes used instead of 0.1% formic acid in both solvents. For more difficult separations, MeCN was used instead of MeOH in both mobile phases as specified herein. Specific gradient conditions are provided in the Examples below, but the following are representative: T(0)→T(3 min) was run isocratically with 10-50% solvent B depending on compound polarity, followed by a gradient to 100% solvent B for 12 min, with the final 5 min at 100% solvent B.

[0140] LCMS analysis was performed on an Agilent instrument. Liquid chromatography was performed on a Phenomenex Kinetex C18 column (2.6 μm; 100 Å; 3X30 mm) at a flow rate of 1.5 mL / min (RT) with UV detection at 220 nm and 254 nm. The mobile phase consisted of solvent A (95% H2O / 5% MeOH / 0.1% AcOH) and solvent B (95% MeOH / 5% H2O / 0.1% AcOH) using the following gradient: T(0) 100% A → T(0.5 min) 100% B → isocratic 100% B → T(2 min). MS detection was performed in parallel using APCI detection in both positive and negative modes.

[0141] Unless otherwise indicated, all numerical values ​​expressing amounts of ingredients, reaction conditions, concentrations, properties, stability, and the like used in the specification and appended claims are understood to be modified in all instances by the word "about." At the very least, each numerical parameter should be construed in light of, at least, the number of reported significant digits and by applying ordinary rounding techniques. Thus, unless indicated to the contrary, the numerical parameters set forth in the specification and appended claims are approximations that may vary depending on the properties sought to be obtained. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the embodiments are approximations, the numerical values ​​set forth in the specific examples are reported as precisely as possible. However, any numerical value inherently contains certain errors resulting from variations in experiments, testing measurements, statistical analyses and the like.

[0142] Examples of synthesis, biological activity, and properties include:

[0143] All compounds described herein were prepared according to the methods shown in Tables 2-5. Mass spectrometry and NMR characterization data are provided for each of the examples. Compounds were tested in the assays described in the Biological Experimental Section. The conventions used for reporting biological data are provided as footnotes to each table.

[0144] General synthesis method A:

[0145] Commercially available 2,6-difluoro-3-nitrobenzoic acids A-1 (Scheme A below) can be converted to carbamates A-2 via a Curtius reaction according to the procedure described in J. Med. Chem. 2003, 46, 1905. Catalytic hydrogenolysis of nitroarenes A-2 using hydrogen gas and a catalyst such as palladium metal on carbon or palladium hydroxide on carbon (Pearlman's catalyst) gave anilines A-3. Aniline A-3 can be deprotected to aniline salt, e.g. compound A-5, using a strong acid (e.g., a solution of anhydrous hydrochloric acid in dioxane) in the presence of a sulfonylating agent, e.g., a sulfonyl chloride, and an organic base, e.g., pyridine, which can be used as a solvent, in the presence or absence of a catalyst, e.g., 4-dimethylaminopyridine, and in the presence of an additional solvent, e.g., dichloromethane or tetrahydrofuran, to give sulfonamide intermediate A-4. Alternatively, 2,6-difluoroaniline A-6 can be converted to its acetanilide A-7 using an acetylating agent, e.g., acetic anhydride, and then converted to monoprotected dianiline A-8, as described in WO2012 / 101238A1. Sulfonylation to sulfonamide A-9 is accomplished under similar conditions used to convert carbamate A-3 to sulfonamide A-4 using a sulfonylating reagent in the presence of an organic base, such as pyridine, with or without a catalyst, such as 4-dimethylaminopyridine, and a solvent, such as dichloromethane or tetrahydrofuran. Treatment of acetanilide A-9 with aqueous hydrochloric acid in the presence of a co-solvent, such as an alcohol, provides the aniline salt A-5.

[0146] Scheme A

[0147] [ka]

[0148] 8-Chloro-2-(methylthio)pyrimidopyrimidine A-10 is commercially available or can be prepared as described in WO2012 / 101238A1. Inhibitors of general formula I are prepared by nucleophilic substitution between compound A-10 and aniline derivative A-5 along a similar procedure to that described in WO2012 / 101238A1. Inhibitors of general formula II are prepared from inhibitors of general formula I by the following two-step procedure, which first generally involves oxidizing the thiomethyl group to the corresponding mixture of methyl sulfoxide and methyl sulfone, which is then reacted with a nucleophile (such as a primary or secondary amine, alcohol, phenol, or NH-containing heterocycle). The latter step is usually carried out in a solvent such as DMSO or NMP in the presence of a base (eg an organic base such as DIEA, trimethylamine, pyridine, etc.) at a temperature in the range of 70-140°C.

[0149] General synthesis method B:

[0150] An alternative method for the present inhibitor is described in Scheme B below. As shown in Scheme B, intermediate A-2 (prepared as described in Scheme A and described in J.Med.Chem.2003,46,1905) is converted to nitroaniline hydrochloride B-1 by cleavage of carbamate protecting group under acidic conditions (e.g., HCl in dioxane or TFA). Chloropyrimidopyrimidine A-10 (prepared as described in Scheme A (WO2012 / 101238A1)) undergoes nucleophilic displacement with aniline salt, e.g., compound B-1, or preferably with aniline free base under conditions similar to those described in WO2012 / 101238A1 to provide intermediate B-2. The nitro functionality of intermediate B-2 is then reduced to the corresponding aniline B-3 by methods well established and familiar to those skilled in the art, for example, using tin(II) chloride, iron or zinc powder in a solvent such as MeOH, EtOH or EtOAc under acidic conditions at temperatures ranging from 50° C. to 100° C. The aniline B-3 is then sulfonylated under basic conditions with a sulfonyl chloride as described in Scheme A to provide the inhibitor of general formula I. The inhibitor of formula I is then converted to the inhibitor of general formula II as described in Scheme A.

[0151] Scheme B

[0152] [ka]

[0153] General synthesis method C:

[0154] An alternative method for preparing the inhibitors using compounds of general formula III and IV is described in Scheme C below. As shown in Scheme C, intermediate B3 prepared as described in Scheme B is converted to chlorosulfonylaniline C-1 by treating with sulfuryl chloride in the presence of an organic base, such as a tertiary amine (e.g., trimethylamine, DIEA, etc.). Intermediate C-1 is reacted with a primary or secondary amine, such as a pyrrolidine derivative, to provide inhibitors of general formula III. Inhibitors of general formula IV are then obtained by the following two-step procedure, which includes oxidizing thiomethyl to a combination of sulfoxide and sulfone, followed by reaction with a nucleophile as described in Scheme A.

[0155] Scheme C

[0156] [ka]

[0157] General synthesis method D:

[0158] Examples of inhibitors of general formula V were prepared according to the sequence shown in Scheme D below.

[0159] Scheme D

[0160] [ka]

[0161] 2,4,8-Trichloropyrimidopyrimidine D-1 was prepared by the procedure described in ACS Med. Chem. Lett. 2011, 2,538 and converted to 4-amino-2,8-dichloropyrimidopyrimidine D-2 by treatment with ammonia as described in WO2010 / 026262A1. Dichloropyrimidopyrimidine D-2 underwent regioselective nucleophilic displacement with aniline salt of general formula A-5 under the general conditions described in Scheme A to provide 8-chloropyrimidopyrimidine intermediate D-3. Chloropyrimidopyrimidine D-3 was displaced a second time with a nucleophile, e.g., a secondary amine or an NH-containing heterocycle, along a protocol similar to that described in WO2012 / 101238A1 to provide inhibitors of general formula V. The latter step is typically carried out in a solvent, such as DMSO or NMP, in the presence of a base (e.g., an organic base, such as DIEA, trimethylamine, pyridine, etc.) at a temperature ranging from 70 to 140° C. Alternatively, intermediate D-3 is reacted with a nucleophile, such as a secondary amine or an NH-containing heteroaryl (e.g., imidazole, benzimidazole, etc.), in the presence of an organic ligand (e.g., racemic BINOL) and an inorganic base under copper-catalyzed cross-coupling conditions. These reactions are typically carried out in a solvent, such as DMSO or NMP, at a temperature ranging from 80° C. to 140° C. Other methods of coupling chloropyrimidines to such nucleophiles are known to those skilled in the art as involving metal-catalyzed processes and can be used to obtain inhibitors of general formula V.

[0162] General synthesis method E:

[0163] Inhibitors of general formula VI are prepared as described in scheme E below. An intermediate of general formula I is first prepared according to general method A or B and then oxidized to a mixture of methyl sulfoxide and methyl sulfone as described in general method A. Intermediate I is then coupled to a 3-indole carboxylic ester (e.g., methyl ester, X=CH) or a 3-indazole carboxylic ester (e.g., methyl ester, X=N) according to a protocol similar to that described in WO2012 / 101238A1. The latter step is usually carried out in a solvent such as THF, DMSO or NMP in the presence of a base (e.g., an organic or inorganic base such as Cs2CO3, KOtBu, DIEA, trimethylamine, pyridine, etc.) at a temperature ranging from ambient temperature to 140°C. Deprotection of the ester protecting group using an inorganic base (e.g., NaOH or KOH) in a mixture of water and a miscible organic solvent (e.g., methanol, ethanol, THF, dioxane, etc.) at temperatures ranging from ambient temperature to 100° C., followed by acidification with a mineral acid (e.g., aqueous hydrochloric acid or aqueous sulfuric acid), an inorganic salt solution (e.g., aqueous NH4Cl or aqueous KHSO4), or an organic acid (e.g., aqueous citric acid or aqueous acetic acid) provided the corresponding carboxylic acid intermediate E-1. Coupling of intermediate E-1 with amines using standard amide coupling reagents (e.g., TBTU, HATU, DCC, EDC, etc.) provided the amide derivatives of general formula VI.

[0164] Scheme E

[0165] [ka]

[0166] General synthesis method F:

[0167] According to the general procedure described in Scheme F below, thiomethyl intermediate I, prepared as described in General Method A or B, was oxidized in step 1 to a mixture of methyl sulfoxide and methyl sulfone as described in General Method A. Condensation with an amine in a solvent, such as THF, in the presence of an organic base (e.g., DIEA, trimethylamine, etc.), prepared as described in Org.Lett. 2011, 13, 3588, gave 3-indolesulfonamide intermediate F-1. In step 2, condensation with an amine in a solvent, such as THF, in the presence of an organic base (e.g., DIEA, trimethylamine), prepared separately as described in Org.Lett. 2011, 13, 3588, gave 3-indolesulfonamide intermediate F-1. Alternatively, N-tosyl-protected indole-3-sulfonyl chloride (prepared by the procedure described in Chemical and Pharmaceutical Bulletin 2009, 57, 591) is reacted with a primary or secondary amine in the presence of a tertiary base, such as DIEA or triethylamine, in a solvent, such as THF, to remove the tosyl protecting group upon treatment with an aqueous inorganic base, such as KOH, to provide intermediate sulfonamide F-1. Intermediate F-1 is then condensed with the oxidized mixture of intermediate I from step 1 following a protocol similar to that described in WO 2012 / 101238 A1. The latter step is typically carried out in a solvent, such as DMSO or NMP, in the presence of a base (e.g., an organic base, such as DIEA, trimethylamine, pyridine, etc.) at a temperature ranging from 70 to 140° C. to provide inhibitors of general formula VII.

[0168] Scheme F

[0169] [ka]

[0170] General synthesis method G:

[0171] 3-Indole thiocyanate G-1 (prepared according to the procedure described in Phosphorus, Sulfur and Silicon and the Related Elements 2014, 189, 1378) was reduced to the corresponding sulfide salt using a reducing agent, e.g., sodium sulfide anhydrate, and directly alkylated with an alkyl halide without isolation to provide sulfide intermediate G-2. Sulfide intermediate G-2 was then converted to sulfone intermediate G-3 using an oxidizing agent, e.g., 3-chloroperoxybenzoic acid. The final inhibitors of general structure VIII were then obtained by the usual methods as described.

[0172] [ka]

[0173] General synthesis method H:

[0174] A bright red solution of commercially available 3-fluoro-2-nitroaniline H-1 is reacted with a primary or secondary amine in the presence of an inorganic base, such as potassium carbonate, or an organic base, such as DIEA, in a solvent, such as MeCN, DMSO or NMP, and heated to a temperature of 40°C to 120°C under thermal or microwave conditions to provide intermediate H-2. Reduction of the nitro group of intermediate H-2 can be achieved using a metal, such as Fe or Zn, in the presence of ammonium chloride, in an alcoholic solvent, such as isopropanol, at a temperature of 40°C to 80°C. The 1,2-phenylenediamine intermediate is then directly converted to the desired benzimidazole intermediate H-3 upon heating with formic acid at a temperature of 40°C to 80°C. The final inhibitor of general structure IX is then obtained from benzimidazole H-3 and compound I by heating under the usual conditions as previously described.

[0175] [ka]

[0176] General synthesis method I:

[0177] Nitroanilines H-2 obtained as described under general synthetic method H are reduced to 1,2-phenylenediamines I-1 in the presence of ammonium chloride using metals such as Fe or Zn in an alcoholic solvent, such as isopropanol, at temperatures between 40° C. and 80° C. The 1,2-phenylenediamine intermediates I-1 are then converted to the desired benzotriazole intermediates I-2 upon treatment with an inorganic nitrite, such as sodium nitrite, under acidic conditions (e.g., AcOH). The final inhibitors of general structure X are then obtained from intermediates I-2 and compounds I by heating under the usual conditions as previously described.

[0178] [ka]

[0179] Sulfonyl chlorides:

[0180] The following sulfonyl chlorides were obtained from commercial sources and used as received: 44-methoxybenzenesulfonyl chloride, 2,4-dichlorobenzenesulfonyl chloride, 2,4-dimethylbenzenesulfonyl chloride, 2-chlorobenzenesulfonyl chloride, 2-methylbenzenesulfonyl chloride, 4-ethylbenzenesulfonyl chloride, 2-cyanobenzenesulfonyl chloride, 2,4-dimethoxybenzenesulfonyl chloride, 2-trifluoromethylbenzenesulfonyl chloride, 3-chlorobenzenesulfonyl chloride, 3-methylbenzenesulfonyl chloride, 2,3-dichlorobenzenesulfonyl chloride, 3-chloro-2-methylbenzenesulfonyl chloride, and 4-methylbenzenesulfonyl chloride. benzenesulfonyl chloride, 2-bromobenzenesulfonyl chloride, 2-chloro-4-fluorobenzenesulfonyl chloride, 2-chloro-6-fluorobenzenesulfonyl chloride, 2,5-dichlorobenzenesulfonyl chloride, 2,5-dimethylbenzenesulfonyl chloride, 2-chloro-6-methylbenzenesulfonyl chloride, 3-fluoro-2-methylbenzenesulfonyl chloride, 2-chloro-4-methylbenzenesulfonyl chloride, 1,3-benzodioxole-5-sulfonyl chloride, 2-chloro-4-(trifluoromethyl)-benzenesulfonyl chloride, 2-methyl-4-nitrobenzenesulfonyl chloride, 2-(difluoromethyl)benzenesulfonyl chloride.

[0181] Other sulfonyl chlorides were prepared by using or adapting literature procedures, as described below.

[0182] 2-Fluoro-4-methoxybenzenesulfonyl chloride:

[0183] [ka]

[0184] Following the procedure described in EP2752410A1, 2-fluoro-4-methoxyaniline (1.00 g, 7.1 mmol) was dissolved in acetonitrile (25 mL) and concentrated hydrochloric acid (10 mL) was added. The mixture was cooled to 0°C in an ice-salt bath. Then, a solution of NaNO2 (0.59 g, 8.5 mmol) in water (1 mL) was added in portions and the mixture was stirred for 1.5 hours at 0°C (light brown solution with a small amount of white solid in suspension). To the resulting mixture, AcOH (12 mL) was added and, after stirring for 10 minutes at 0°C, NaHSO3 (7.37 g, 10 equiv., 71 mmol) was added. After stirring for 5 min, Cu(II) chloride (0.96 g, 1 equiv.) and CuCl (70 mg, 0.1 equiv.) were added and the green suspension was stirred in an ice bath and allowed to warm to room temperature over 1 h, after which it was stirred at room temperature for an additional 18 h (TLC R f hexane / EtOAc). The reaction mixture was then poured into water (100 mL) and extracted with EtOAc. The extract was washed with water, dried over MgSO4, and filtered through a pad of silica gel (15 mL) using 1:1 hex / EA as eluent. Removal of the volatiles under reduced pressure gave 1.22 g of a clear light brown oil (TLC showed the presence of a more polar unidentified impurity after aqueous work-up). 1 H NMR(CDCl3) δ:7.88(t,J=8.6Hz,1H),6.76~6.93(m,2H),3.93(s,3H). uniformity 1 ~70% by H NMR.

[0185] The following sulfonyl chlorides were prepared using a similar procedure:

[0186] 2-Cyano-4-fluorobenzenesulfonyl chloride: Prepared using 2-cyano-4-fluoroaniline as the starting material. The crude material was highly impure but was successfully used in the sulfonylation reaction.

[0187] 4-Methoxy-2-trifluoromethylbenzenesulfonyl chloride: Prepared using 4-methoxy-2-trifluoromethylaniline: 1 H NMR(CDCl3) δ:8.30(d,J=9.0Hz,1H),7.42(d,J=2.3Hz,1H),7.19(dd,J=9.0,2.7Hz,1H),3.99(s,3H).

[0188] 4-Chloro-2-methylbenzenesulfonyl chloride:

[0189] [ka]

[0190] It was prepared by chlorosulfonylation of meta-chlorotoluene according to the procedure described in Acta Crystallographica Section E 2009,65(4),o800. Meta-chlorotoluene (1 mL) was dissolved in CHCl3 (4 mL) and the solution was cooled in an ice bath. Chlorosulfonic acid (2.5 mL) was added dropwise over 15 min with slow evolution of HCl gas. Upon completion, the reaction mixture was allowed to warm to room temperature. TLC showed no starting material (Rf=0.8, in 8:2 hex / EA) and a new spot with some tailing (Rf=0.7, in 8:2 hex / EA) was formed. The reaction mixture was poured onto ice (50 mL), DCM (15 mL) was added and the product organic phase was separated, washed with cold water, dried (MgSO4) and concentrated to a colorless oil (0.87 g) which was used without further purification: 1 H NMR(CDCl3) δ:8.01(d,J=8.6Hz,1H),7.43(s,1H),7.40(dd,J=8.6,2.0Hz,1H),2.78(s,3H).

[0191] The following sulfonyl chlorides were prepared using a similar procedure with some modifications, as described below.

[0192] 4-Methoxy-2-methylbenzenesulfonyl chloride:

[0193] [ka]

[0194] 3-Methoxytoluene (6.00 g) was dissolved in CHCl3 (30 mL) and the solution was cooled to -35 °C (bath temperature). Chlorosulfonic acid (15 mL) was added dropwise over 20 min (no significant evolution of HCl / SO2 gas). The clear solution was then stirred at -30 to -25 °C for 15 min (no noticeable evolution of gas). The reaction mixture was carefully poured onto ice (50 mL), DCM (50 mL) was added, and the slightly milky product organic phase was separated, washed with cold water, dried (MgSO4), and concentrated under vacuum to a colorless oil (8.28 g, 76% yield). NMR indicates the presence of a single isomer: 1 H NMR(CDCl3) δ:8.01(d,J=8.6Hz,1H),6.72~6.95(m,2H),3.90(s,3H),2.75(s,3H).

[0195] 2-Chloro-4-methoxybenzenesulfonyl chloride:

[0196] [ka]

[0197] 3-Chloroanisole (1.00 g) was dissolved in CHCl3 (4 mL) and the solution was cooled to approximately -35°C. Chlorosulfonic acid (2.5 mL) in CHCl3 (2 mL) was added dropwise over 15 min. Upon completion, there was only baseline material by TLC (SM Rf = 0.8 in 8:2 hex / EA). Upon warming the reaction mixture to room temperature, gas evolution was observed and isomeric products were observed by TLC (Rf = 0.30 and 0.25 in 8:2 hex / EA). After stirring at room temperature for 30 min, a white precipitate began to form. The reaction mixture was poured onto ice (50 mL), DCM (15 mL) was added, and the organic product phase was separated, washed with cold water, dried (MgSO4), and concentrated to a colorless oil that crystallized on standing as white needles (0.89 g). 1 H NMR showed a mixture of two isomers in a 60:40 ratio, which were separated by flash chromatography on silica gel using 8:2 hexanes / EtOAc as eluent. The desired isomer (more polar): 1 H NMR(CDCl3) δ:7.90(d,J=8.6Hz,1H),7.04~7.19(m,2H),4.08(s,3H).

[0198] 2,3-Dimethylbenzenesulfonyl chloride:

[0199] [ka]

[0200] It was isolated as the minor isomer following chlorosulfonylation of o-xylene as described in WO 2003 / 055478. 1 H NMR(CDCl3) δ:7.96(d,J=7.8Hz,1H),7.52(d,J=7.4Hz,1H),7.30(t,J=7.8Hz,1H),2.71(s,3H),2.41(s,3H).

[0201] 3-Fluoro-2-methyl-4-methoxybenzenesulfonyl chloride:

[0202] [ka]

[0203] Step 1: To a solution of 2-fluoro-3-methylphenol (4.32 mL, 39.7 mmol) in acetone (50 mL) was added potassium carbonate (6.58 g, 47.6 mmol) followed by iodomethane (2.75 mL, 43.7 mmol). The reaction mixture was then refluxed at 60° C. overnight. The reaction mixture was then cooled to room temperature, filtered (washed with 2×10 mL acetone), and concentrated under reduced pressure. The crude product was extracted with water (30 mL) and EtOAc (2×50 mL). The organic layer was then separated, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel using 0-5% EtOAc / hexane to give 2-fluoro-3-methylanisole as a clear colorless liquid (5.30 g, 95% yield): 1 H NMR(CDCl3) δ:6.95(td,J=8.0,1.4Hz,1H),6.85~6.71(m,2H),3.87(s,3H),2.28(d,J=2.3Hz,3H).

[0204] Step 2: To a solution of 2-fluoro-3-methylanisole from step 1 (1.00 g, 7.13 mmol) in DCM (5.6 mL) was added a solution of chlorosulfonic acid (1.13 mL, 16.5 mmol) in DCM (5.6 mL) over 5 min. The light brown reaction mixture containing a viscous liquid layer was stirred at room temperature for 10 min and then quenched by pouring into water (10 mL) and ice (5 g). The aqueous layer was extracted with DCM (2x10 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the desired sulfonyl chloride (1.70 g, 100% yield) as a colorless liquid: 1H NMR(CDCl3) δ:7.87(dd,J=9.0,1.8Hz,1H),6.97~6.86(m,1H),3.97(s,3H),2.66(d,J=2.8Hz,3H).

[0205] 3-Chloro-2-methyl-4-methoxybenzenesulfonyl chloride:

[0206] [ka]

[0207] Following a similar procedure as for 3-fluoro-2-methyl-4-methoxybenzenesulfonyl chloride (step 1), starting from 2-chloro-3-methylphenol, 2-chloro-3-methylanisole was obtained in quantitative yield as a colorless liquid: 1 H NMR(CDCl3) δ:7.12(t,J=7.9Hz,1H),6.87~6.83(m,1H),6.79(d,J=8.2Hz,1H),3.89(s,3H),2.38(s,3H).

[0208] Treatment with chlorosulfonic acid as described for 3-fluoro-2-methyl-4-methoxybenzenesulfonyl chloride (step 2) afforded the desired 3-chloro-2-methyl-4-methoxybenzenesulfonyl chloride in 96% yield as a colorless liquid: 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).

[0209] 2-Ethylbenzenesulfonyl chloride:

[0210] [ka]

[0211] 2-Ethylbenzenethiol (1.46 mL, 10.3 mmol) and KCl (776 mg, 10.3 mmol) were dissolved in water (38 mL) and oxone (登録商標) (15.8 g, 25.8 mmol) was added in small portions. After stirring for 1 h at room temperature, the reaction was deemed complete by LCMS analysis and the reaction mixture was extracted with EtOAc (4x5 mL). The extracts were dried (Na2SO4) and concentrated under reduced pressure to give a white crystalline solid (1.43 g, 68% yield) which was used as is: 1 H NMR(CDCl3) δ:8.07(dd,J=8.1,1.3Hz,1H),7.66(td,J=7.6,1.3Hz,1H),7.49(d,J=7.7Hz,1H),7.45~7.38(m,1H),3.20(q,J=7.5Hz),1.36(t,J=7.5Hz).

[0212] 3-Fluoro-2-ethylbenzenesulfonyl chloride:

[0213] [ka]

[0214] Step 1: 2-Bromo-6-fluorobenzaldehyde (6.00 g, 29.5 mmol) was dissolved in dry THF (60 mL) and the solution was cooled to -78 °C under argon atmosphere. Methylmagnesium bromide (3.0 M solution in diethyl ether, 13.4 mL, 40.3 mmol) was added dropwise and the mixture was stirred at -78 °C for 30 min. The reaction was then quenched with 10% hydrochloric acid (50 mL) and the product was extracted into ether (2x50 mL). The extracts were dried (MgSO4) and concentrated and the residue was purified by Combiflash on silica gel using 10-30% EtOAc in hexanes as eluent. (登録商標) to give the desired alcohol derivative as a colorless oil (6.20 g, 96% yield): 1H NMR(CDCl3) δ:7.35(ddd,J=7.9,3.1,2.0Hz,1H),7.16~6.99(m,2H),5.35(q,J=6.7Hz,1H),1.61(dd,J=6.8,1.1Hz,3H).

[0215] Step 2: Indium(III) chloride (412 mg, 1.83 mmol) was suspended in DCM (40 mL) and chlorodiisopropylsilane (8.42 mL, 49.3 mmol) was added. The alcohol from step 1 (4.00 g, 18.3 mmol) in DCM (8 mL) was added and the mixture was stirred at room temperature for 3 h to give a clear solution. The reaction mixture was quenched with water (50 mL), extracted with diethyl ether (3x20 mL), washed with brine and dried (MgSO4). Concentration and purification by flash chromatography using hexane as eluent provided the silylated ether of the starting alcohol.

[0216] The material was dissolved in DCE (41 mL) and chlorodiisopropylsilane (0.78 mL, 4.6 mmol) and indium(III) chloride (103 mg, 4.6 mmol) were added. The mixture was stirred at 80° C. for 3 h. After cooling to ambient temperature, the reaction mixture was diluted with hexane (100 mL), washed with water (100 mL), and the aqueous layer was back-extracted with hexane (2×50 mL). The combined organic phases were dried (Na2SO4) and concentrated to give a colorless oil, which was purified by flash chromatography on silica gel using hexane as eluent to give 2-bromo-6-fluoro-ethylbenzene (3.71 g, 100%) as a colorless oil: 1 H-NMR(CDCl3) δ:7.32(d,J=7.8Hz,1H),7.11~6.91(m,2H),2.82(dq,J=7.5,2.2Hz,2H),1.20~1.15(m,3H).

[0217] Step 3: The aryl bromide from step 2 (3.71 g, 18.3 mmol) was dissolved in toluene (60 mL) and N,N-diisopropylethylamine (6.40 mL, 36.5 mmol) was added. The solution was then degassed by three cycles of evacuation and back-filling with nitrogen. Tris(dibenzylideneacetone)dipalladium(0) (836 mg, 0.9 mmol), 4,4-bis(diphenylphosphino)-9,9-dimethylxanthene (1.08 g, 1.83 mmol) and 2-ethylhexyl-3-mercaptopropionate (4.59 mL, 19.2 mmol) were added and the mixture was degassed twice before refluxing under nitrogen overnight. The reaction mixture was then cooled to room temperature, quenched with water (50 mL) and extracted with EtOAc (2x50 mL). The combined organic phases were washed with 10% aqueous hydrochloric acid (75 mL) and dried (Na2SO4). Concentration under reduced pressure and purification of the residue by flash chromatography using 0-15% EtOAc in hexanes as eluent gave the desired sulfide intermediate (4.00 g) as an orange oil contaminated with some unreacted starting thiol. This material was used as is in the next step.

[0218] Step 4: The crude sulfide derivative from step 3 (estimated 4.00 g, 11.7 mmol) was dissolved in THF (41 mL) and potassium tert-butoxide (1.0 M in THF, 14.1 mL, 14.1 mmol) was added dropwise. The resulting solution was stirred at room temperature for 1 h. The reaction was then quenched by the addition of saturated aqueous NH4Cl (40 mL) and extracted with EtOAc (2x30 mL). The combined organic phase was concentrated and the deep orange residue was passed through a small pad of silica gel using hexanes for washing to give a mixture of thiol and disulfide (1.50 g) which was used as is in the next step (Caution: foul odor).

[0219] Step 5: The crude mixture of thiophenol and disulfide from step 4 (assumed to be 1.50 g, 9.6 mmol) and KCl (723 mg, 9.6 mmol) were suspended in water (40 mL) and oxone (登録商標) (14.8 g, 24 mmol) was added portionwise. After stirring for 1 h at room temperature, the reaction mixture was extracted with EtOAc (2x20 mL) and the extracts were dried (Na2SO4) and concentrated under reduced pressure to give the crude sulfonyl chloride, which was used directly to prepare the corresponding fragment A-5 and aniline A-8 (see Table 1 below).

[0220] 3-Chloro-2-ethylbenzenesulfonyl chloride:

[0221] [ka]

[0222] The sulfonyl chloride was prepared following the same procedure as 3-fluoro-2-ethylbenzenesulfonyl chloride, but starting from 2-bromo-6-chlorobenzaldehyde.

[0223] Step 1 (98% yield, as a white solid): 1 H NMR(CDCl3) δ:7.49(dd,J=8.0,1.2Hz,1H),7.33(dd,J=8.0,1.2Hz,1H),7.05(t,J=8.0Hz,1H),5.58(q,J=6.9Hz,1H),1.64(d,J=6.9Hz,3H).

[0224] Step 2 (100% yield, as a colorless oil): 1 H-NMR (CDCl3) δ:7.43(dd,J=8.0,1.2Hz,1H),7.29(dd,J=8.0,1.2Hz,1H),6.96(t,J=8.0Hz,1H),2.97(q,J=7.5Hz,2H),1.17(t,J=7.5Hz,3H).

[0225] Step 3 (81% yield, as an orange oil):1 H-NMR(CDCl3) δ:7.24~7.18(m,2H),7.08(t,J=7.9Hz,1H),4.07~3.96(m,2H),3.17(t,J=7.4Hz,2H),2.96(q,J=7.5Hz,2H),2.6 4(t,J=7.4Hz,2H),1.56(dd,J=11.9,5.8Hz,2H),1.40~1.21(m,9H),1.16(t,J=7.5Hz,3H),0.89(t,J=7.4Hz,6H).

[0226] Step 4 (99% yield, as a colorless liquid): 1 H-NMR (CDCl3) δ:7.15(d,J=7.9Hz,2H),6.97~6.92(m,1H),3.41(s,1H),2.87(q,J=7.5Hz,2H),1.18(t,J=7.5Hz,3H).

[0227] Step 5 (crude material, used without purification): 1 H-NMR (CDCl3) δ:8.03(dd,J=8.2,1.3Hz,1H),7.73(dd,J=8.0.1.3Hz,1H),7.37(t,J=8.1Hz,1H),3.30(q,J=7.4Hz,2H),1.33(t,J=7.4Hz,3H).

[0228] 2-Methyl-3-(trifluoromethyl)benzenesulfonyl chloride:

[0229] [ka]

[0230] The sulfonyl chloride was prepared following the same procedure as 3-fluoro-2-ethylbenzenesulfonyl chloride, but starting from commercially available 2-methyl-3-(trifluoromethyl)bromobenzene.

[0231] Step 3 (100% yield, as an orange oil): 1H-NMR(CDCl3) δ:7.49(d,J=7.9Hz,2H),7.26~7.19(m,1H),4.06~4.00(m,2H),3.18(dd,J=9.1,5.6Hz,2H),2.6 5(dd,J=9.2,5.6Hz,2H),2.50(d,J=1.3Hz,3H),1.33~1.23(m,11H),0.88(td,J=7.4,2.3Hz,6H).

[0232] Step 4 (quantitative yield, as a colorless oil): 1 H-NMR (CDCl3) δ:7.43(dd,J=7.9,2.2Hz,2H),7.12(t,J=7.8Hz,1H),3.42(s,1H),2.43(s,3H).

[0233] Step 5 (quantitative yield, as a white solid): 1 H-NMR (CDCl3) δ:8.31(d,J=8.1Hz,1H),8.00(t,J=7.7Hz,1H),7.55(dd,J=15.6,7.6Hz,1H),2.93(s,3H).

[0234] General procedure for the preparation of sulfonyl chlorides from aryl bromides:

[0235] [ka]

[0236] Step 1: Aryl bromide 1 (1.00 mmol) was diluted in toluene (1.70 mL). The mixture was degassed for 5 min by bubbling nitrogen through the solution. Tris(dibenzylideneacetone)-dipalladium(0) (0.02 mmol), 4,4-bis(diphenylphosphino)-9,9-dimethylxanthene (0.04 mmol) and N,N-diisopropylethylamine (2.0 mmol) were added and the mixture was then degassed again for 5 min. Benzyl mercaptan (1 mmol) was then added and the resulting mixture was heated at reflux overnight (oil bath, T=115° C.). After completion, the reaction was cooled to room temperature, diluted with EtOAc (20 mL) and quenched with H2O (20 mL). The aqueous layer was extracted with EtOAc (2×20 mL). The combined organic layers were washed with brine (50.0 mL), dried (Na2SO4), filtered, and concentrated under reduced pressure. The crude product was further purified by flash chromatography (0-10% EtOAc in hexanes, 35 mL / min, product eluted with 100% hexanes). Fractions of interest were collected and concentrated under reduced pressure to give the title compound 2.

[0237] Step 2: Compound 2 (1.00 mmol) was dissolved in acetic acid (1.90 mL) and H2O (0.65 mL) was added to give a heterogeneous solution. N-chlorosuccinimide (4.00 mmol) was added portionwise. The reaction was stirred and monitored by LCMS (sample was quenched with N-methylpiperazine). After completion of the reaction, the mixture was concentrated under reduced pressure. The resulting mixture was poured slowly into saturated aqueous NaHCO3, which caused gas evolution. The mixture was extracted with EtOAc (2x75 mL). The combined organic layers were washed with brine, dried (Na2SO4), filtered, and concentrated under reduced pressure. The crude compound was further purified by normal phase chromatography (0-40% EtOAc in hexanes, 60 mL / min, product came out with 100% hexanes). Fractions of interest were collected and concentrated under reduced pressure to give the title compound 3.

[0238] 2-Chloro-3-methylbenzenesulfonyl chloride:

[0239] [ka]

[0240] The sulfonyl chloride was prepared following the general procedure starting from commercially available 1-bromo-2-chloro-3-methylbenzene.

[0241] Benzyl (2-chloro-3-methylphenyl) sulfide: yellow solid, 51% yield, 95% purity (at 220 nm). (ES - )MH=247.2; 1 H NMR (400MHz, CDCl3) δ7.39~7.35(m,2H),7.33~7.28(m,2H),7.28~7.23(m,1H+CDCl3),7.11~7.03(m,3H),4.15(s,2H),2.38(s,3H).

[0242] 2-Chloro-3-methylbenzenesulfonyl chloride: Light yellow oil, 70% yield, 60% purity (at 254 nm). LCMS: LCMS sample was quenched with N-methylpiperazine (resulting sulfonamide, MW=288.8), (ES + ) M+H=289.2. Used crude.

[0243] 3-Fluoro-2-(trifluoromethyl)benzenesulfonyl chloride:

[0244] [ka]

[0245] The sulfonyl chloride was prepared following the general procedure starting from commercially available 1-bromo-3-fluoro-2-(trifluoromethyl)benzene.

[0246] Benzyl (3-fluoro-2-(trifluoromethyl)phenyl) sulfide: yellow oil, 66% yield, 98% purity (at 254 nm). (ES - )MH=285.2.

[0247] 3-Fluoro-2-(trifluoromethyl)benzenesulfonyl chloride: pale yellow oil, 93% yield, 98% purity (at 254 nm) LCMS: LCMS sample was quenched with N-methylpiperazine (resulting sulfonamide MW=326.1), (ES + )M+H=327.1.

[0248] 3-Chloro-2-(trifluoromethyl)benzenesulfonyl chloride:

[0249] [ka]

[0250] The sulfonyl chloride was prepared following the general procedure starting from commercially available 1-bromo-3-chloro-2-(trifluoromethyl)benzene.

[0251] Benzyl (3-chloro-2-(trifluoromethyl)phenyl) sulfide: White solid, 59% yield, 90% purity (at 220 nm). (ES - )MH=301.2; 1 H NMR (400MHz, CDCl3) δ7.46~7.14(m,8H+CDCl3),4.16(s,2H).

[0252] 3-Chloro-2-(trifluoromethyl)benzenesulfonyl chloride:

[0253] Colorless oil, 66% yield. LCMS: LCMS sample was quenched with N-methylpiperazine (resulting sulfonamide, MW=343.5), (ES + ) M+H=343.2. Used crude.

[0254] 3,4-Difluoro-2-methylbenzenesulfonyl chloride:

[0255] [ka]

[0256] The sulfonyl chloride was prepared following the general procedure starting from commercially available 1-bromo-3-chloro-2-(trifluoromethyl)benzene.

[0257] Benzyl (3,4-difluoro-2-methylphenyl) sulfide: orange oil, 96% yield, 96% purity (at 254 nm). (ES - )MH=249.2; 1 H NMR (400 MHz, DMSO-d6) δ 7.32-7.17 (m, 7H), 4.16 (s, 2H), 2.18 (d, J = 2.7 Hz, 3H). 3,4-Difluoro-2-methylbenzenesulfonyl chloride: pale yellow oil, 49% yield. LCMS: LCMS sample was quenched with N-methylpiperazine (resulting sulfonamide, MW = 290.3), (ES + )M+H=291.2; 1 H NMR (400MHz, DMSO-d6) δ7.56(ddd,J=8.4,5.5,1.8Hz,1H),7.15(dd,J=18.4,8.4Hz,1H),2.46(d,J=2.8Hz,3H).

[0258] 2,4-Dimethyl-3-fluorobenzenesulfonyl chloride:

[0259] [ka]

[0260] The sulfonyl chloride was prepared following the general procedure starting from commercially available 1-bromo-2,4-dimethyl-3-fluorobenzene.

[0261] Benzyl (3-fluoro-2,4-dimethylphenyl) sulfide: orange oil, 95% crude yield, 80% purity (at 254 nm). Used as crude.

[0262] 3-Fluoro-2,4-dimethylbenzene-1-sulfonyl chloride: orange oil, 89% yield of crude product, 74% purity (at 254 nm). LCMS: LCMS sample was quenched with N-methylpiperazine (resulting sulfonamide, MW=286.4), (ES + ) M+H=287.1. Used crude.

[0263] 2-Methylpyridine-3-sulfonyl chloride:

[0264] [ka]

[0265] The sulfonyl chloride was prepared following the general procedure starting from commercially available 3-bromo-2-methylpyridine.

[0266] 3-(Benzylthio)-2-methylpyridine: Orange liquid, 88% yield, 94% purity (at 220 nm). (ES + )M+H=215.8,(ES - )MH=214.1. 1 H NMR (400MHz, DMSO-d6) δ8.30(dd,J=4.9,1.5Hz,1H),7.57~7.45(m,1H),7.31~7.27(m,5H),7.07(dd,J=7.6,5.1Hz,1H),4.09(s,2H),2.58(s,3H).

[0267] 2-Methylpyridine-3-sulfonyl chloride: pale yellow oil, 100% yield, 95% purity (at 254 nm). LCMS sample was diluted with HO (resulting sulfonic acid, MW=173.1) (ES - )MH=171.9; 1H NMR (400MHz, CDCl3) δ8.80(dd,J=4.8,1.6Hz,1H),8.33(dd,J=8.1,1.7Hz,1H),7.43~7.36(m,1H),3.02(s,3H).

[0268] 6-Methoxy-4-methylpyridine-3-sulfonyl chloride:

[0269] [ka]

[0270] Preparation of 2-ethylhexyl 3-((6-methoxy-4-methylpyridin-3-yl)thio)propanoate (2): 5-Bromo-2-methoxy-4-methylpyridine (6.00 g, 29.7 mmol) was dissolved in toluene (100 mL) and N,N-diisopropylethylamine (10.4 mL, 59.4 mmol) was added. The mixture was degassed for 5 minutes by bubbling nitrogen through the solution. Tris(dibenzylideneacetone)dipalladium(0) (1.36 g, 1.49 mmol), 4,4-bis(diphenylphosphino)-9,9-dimethylxanthene (1.75 g, 2.97 mmol) and 2-ethylhexyl-3-mercaptopropionate (7.47 mL, 31.2 mmol) were added. The mixture was again degassed for 5 minutes. The mixture was heated at reflux (oil bath, T=117° C.) overnight. The reaction was cooled to room temperature, diluted with EtOAc (100 mL), and quenched with H2O (100 mL). The organic and aqueous layers were separated, and the aqueous layer was extracted with EtOAc (2×50.0 mL). The combined organic layers were washed with HCl (10% in H2O, 50.0 mL), dried (Na2SO4), filtered, and concentrated under reduced pressure. The crude product was purified by flash chromatography (330 g silica gel column, EtOAc-hexanes, 0-20%) to give the title compound as an orange oil (10.0 g, 99% yield). (ES + )M+H=340.2; 1H NMR(400MHz, CDCl3) δ8.19(s,1H),6.64(s,1H),3.99(dd,J=5.9,2.4Hz,2H),3.93(s,3H),2.96(t,J=7.3Hz,2H),2.55(t ,J=7.3Hz,2H),2.42(d,J=0.5Hz,3H),1.56(dt,J=12.1,6.0Hz,1H),1.39~1.22(m,8H),0.88(m,6H).

[0271] Preparation of 6-methoxy-4-methylpyridine-3-thiol (3): To a −78° C. solution of compound 2 (10.0 g, 29.5 mmol) in THF (105 mL) was added potassium t-butoxide (1.00 M in THF, 35.3 mL, 35.3 mmol) dropwise and a precipitate formed. The resulting suspension was stirred at −78° C. for 30 min. The reaction was quenched by the addition of NH4Cl (50.0 mL) and the mixture was extracted with CHCl (2×50.0 mL). The combined organic layers were dried (Na2SO4), filtered, and concentrated under reduced pressure to give a dark orange liquid. The crude product was purified by flash chromatography (100% hexanes) to give a mixture of the title compound and its disulfide (4.56 g), which was used in the next step without further purification. Thiol 3: (ES + ) M+H=156.9, and disulfide: R t =1.92 minutes, (ES + )M+H=309.0.

[0272] Preparation of 6-methoxy-4-methylpyridine-3-sulfonyl chloride (4): To a mixture of thiol 3 (4.56 g, 29.4 mmol) in HO (123 mL) was added potassium chloride (2.21 g, 29.3 mmol), followed by portionwise addition of oxone (45.2 g, 73.5 mmol). After completion of the reaction (1 h), the mixture was extracted with EtOAc (2×20.0 mL) and the combined organic layers were dried (NaSO) and concentrated under reduced pressure. The crude product obtained was used without any further purification.

[0273] 3-Methylpyridine-4-sulfonyl chloride:

[0274] [ka]

[0275] Step 1: 4-Bromopyridine (1.00 mmol) was dissolved in toluene (1.70 mL) and N,N-diisopropylethylamine (2.00 mmol) was added. The mixture was degassed for 5 min by bubbling nitrogen through the solution. Tris(dibenzylideneacetone)-dipalladium(0) (0.02 mmol), 4,4-bis(diphenylphosphino)-9,9-dimethylxanthene (0.04 mmol) and phenylmethanethiol / benzyl mercaptan (1.00 mmol) were added. The mixture was degassed again for 5 min. The mixture was heated at reflux (oil bath, T=115° C.) for 18 h. The reaction was cooled to room temperature, diluted with EtOAc (10.0 mL) and quenched with H2O (10.0 mL). The aqueous and organic layers were separated and the aqueous layer was extracted with EtOAc (2x10.0 mL) and the combined organic phase was washed with HCl (10% in H2O, 10.0 mL), dried (Na2SO4), filtered and concentrated under reduced pressure. The crude product was purified by flash chromatography (EtOAc-hexane, 10%-35%) to give sulfide 2 (90% yield). (ES + )M+H=216.1; 1 H NMR (400MHz, CDCl3) δ8.28(d,J=4.6,1H),8.24(s,1H),7.43~7.27(m,5H),7.20(t,J=5.4Hz,1H),4.25(s,2H),2.28(s,3H).

[0276] Step 2: Compound 2 (1.00 mmol) was dissolved in CH2Cl2 (11.5 mL) and cooled to -10 °C. HCl (1.00 M in H2O, 5.70 mL) was added and stirred at -10 °C for 5 min. Sodium hypochlorite (10% solution in H2O, 3.00 mmol) was added over 10 min, keeping the temperature below 0 °C. The mixture was stirred at 0 °C for 10 min. The organic and aqueous layers were separated. The organic layer was dried (Na2SO4). The crude sulfonyl chloride was used in the next step without further purification or evaporation: LCMS sample was quenched with N-methylpiperazine (resulting sulfonamide, MW = 255.3); (ES + )M+H + =256.2.

[0277] 2,3-Dimethylpyridine-4-sulfonyl chloride:

[0278] [ka]

[0279] The same procedure as for 3-methylpyridine-4-sulfonyl chloride was used starting from 2,3-dimethyl-4-bromopyridine.

[0280] Step 1: 4-(benzylthio)-2,3-dimethylpyridine (2): (92% yield). (ES + )M+H=230.2; 1 H NMR (400MHz, CDCl3) δ:8.17(d,J=5.5Hz,1H),7.42~7.28(m,5H),6.99(d,J=5.5Hz,1H),4.18(s,2H),2.53(s,3H),2.24(s,3H).

[0281] Step 2: 2,3-Dimethylpyridine-4-sulfonyl chloride (3): LCMS sample was quenched with N-methylpiperazine (resulting sulfonamide, MW=269.3); (ES + )M+H+ =270.2.

[0282] Protected sulfonyl chloride for fragment B69:

[0283] [ka]

[0284] Step 1: To a solution of commercially available aminopyridine (1.00 g, 4.27 mmol) and N-benzylcarbamoyl chloride (0.9421 g, 5.5546 mmol) in EtOAc (20 mL) was added 20 mL of a saturated aqueous solution of NaHCO3. The solution was stirred at room temperature for 16 h. Upon completion, EtOAc was added to the reaction mixture and the organic layer was separated, washed with brine, dried over MgSO4, then filtered and concentrated. The residue was adsorbed onto SiO2 and then purified on SiO2 with EtOAc in hexane to give the desired protected aminopyridine (1.00 g, 2.72 mmol, 64%). 1 H NMR (400MHz, DMSO-d6) δ:10.35(s,1H),8.09(d,J=8.61Hz,1H),7.47(d,J=9.00Hz,1H),7.23~7.44(m,4H),5.16(s,2H),2.53(s,3H). MS m / z 369.2(MH + ).

[0285] Step 2: A degassed solution of iodopyridine from step 1 (0.61 g, 1.66 mmol), tris(dibenzylideneacetone)-dipalladium(0) chloroform adduct (86 mg, 0.0828 mmol), 9,9-dimethyl-9h-xanthene-4,5-diyl)bis(diphenylphosphine (96 mg, 0.166 mmol), DIPEA (0.576 mL, 3.31 mmol) and benzyl mercaptan (0.233 mL, 1.99 mmol) in toluene (15 mL) was stirred at 115 °C under N for 3 h. After completion, SiO was added to the reaction mixture and concentrated under vacuum. The residue was purified on a SiO column with EtOAc in hexane to give the expected sulfide (560 mg, 93%). 1 H NMR(400MHz, CDCl3) δ:7.71(d,J=8.61Hz,1H),7.54(d,J=8.61Hz,1H),7.46(br.s.,1H),7.31~7.43(m, 5H),7.19~7.26(m,2H),7.12~7.19(m,2H),5.22(s,2H),3.96(s,2H),2.41(s,3H). MS m / z 365.2(MH + ).

[0286] Step 3: To a solution of the sulfide from step 2 (300 mg, 0.823 mmol) in 90% AcOH in water (16 mL) was added N-chlorosuccinimide (330 mg, 2.47 mmol). The reaction mixture was stirred at room temperature for 3 h. The reaction mixture was evaporated to dryness, then diluted with EtOAc and washed with water followed by brine. The organic layer was dried over MgSO4, filtered and concentrated in vacuo to give the desired sulfonyl chloride B49 (282 mg, 99%), which was used as is: 1 H NMR (400MHz, CDCl3) δ:8.28(d,J=9.00Hz,1H),8.02(d,J=9.00Hz,1H),7.72(br.s.,H),7.34~7.60(m,5H),5.27(s,2H),2.86(s,3H). MS m / z 341.2(MH + ).

[0287] 2,2-Difluorobenzo[d][1,3]dioxole-4-sulfonyl chloride:

[0288] [ka]

[0289] Thionyl chloride (5.96 mL) was added dropwise to water (30 mL) over 20 min, and the mixture was stirred at room temperature for 48 h to produce a sulfur dioxide-containing solution. In a separate vessel, 2,2,-difluorobenzo[d][1,3]dioxol-4-amine (1.00 g, 5.78 mmol) was added dropwise to ice-cold concentrated hydrochloric acid (7 mL) to produce a white precipitate. A solution of sodium nitrite (523 mg, 7.5 mmol) in water (2 mL) was added dropwise to the aniline hydrochloride over 5 min to produce an orange reaction mixture. The orange suspension was then slowly added at 5° C. to the sulfur dioxide solution from above to which 10 mg of cuprous chloride had been added previously. The mixture was stirred in an ice bath for an additional 2 h (gas evolution was observed and an orange liquid was deposited at the bottom of the flask). After completion was determined by LCMS analysis, the reaction mixture was extracted with DCM (2x20 mL), dried (Na2SO4), filtered and gave the desired sulfonyl chloride as an orange oil which was used without further purification (100% crude yield): 1 H-NMR (400MHz, CDCl3) δ7.65(dd,J=8.4,1.1Hz,1H),7.44(dd,J=8.1,1.1Hz,1H),7.32(t,J=8.2Hz,1H).

[0290] 4-Chloro-3-fluoro-2-methylbenzenesulfonyl chloride:

[0291] [ka]

[0292] Preparation of N-(3-fluoro-2-methylphenyl)pivalamide (2): To a solution of 3-fluoro-2-methylaniline (10.9 mL, 93.0 mmol) in THF (240 mL) at 0 °C was added triethylamine (14.9 mL, 106 mmol) followed by pivaloyl chloride (13.1 mL, 105 mmol) over 10 min. The mixture was warmed to room temperature and stirred for 2 h. The volatiles were evaporated under reduced pressure and the residue was partitioned between H2O (250 mL) and EtOAc (150 mL). The organic and aqueous layers were separated. The aqueous layer was extracted with EtOAc (4x60 mL). The combined organic layers were washed with brine (60 mL), dried (Na2SO4) and concentrated under reduced pressure to give the title compound (18.5 g, 95% yield) as a solid. (ES + )M+H=210.2.

[0293] Preparation of N-(4-chloro-3-fluoro-2-methylphenyl)pivalamide (3): To compound 2 (4.20 g, 20.1 mmol) in DMF (50.0 mL) at room temperature was added N-chlorosuccinimide (2.76 g, 20.1 mmol) over 10 min (in three portions). The mixture was heated at 80° C. for 90 min. Additional N-chlorosuccinimide (541 mg, 4.01 mmol) was added and this was stirred at 80° C. for 45 min. The mixture was cooled to room temperature and diluted with EtOAc (30 mL) and water (60 mL). The organic and aqueous layers were separated. The aqueous layer was extracted with EtOAc (3×20 mL). The combined organic layers were washed with H2O (3x30 mL), brine (20.0 mL), dried (Na2SO4), and concentrated under reduced pressure to give the crude compound (5.20 g). The crude product was dissolved in cyclohexane (30 mL) and heated at 45-50 °C until all solids were dissolved. The solution was cooled to room temperature and the precipitated white solid was filtered and washed with cyclohexane (3x5 mL) to give the title compound (1.95 g, 40% yield) as a solid. (ES + )M+H=244.1.

[0294] Preparation of 4-chloro-3-fluoro-2-methylaniline (4): To a solution of compound 3 (1.60 g, 6.57 mmol) in dioxane (18 mL) at room temperature, HCl (6.00 M in water, 23 mL, 138 mmol) was added over 5 min. The mixture was heated at 100 °C for 20 h. The mixture was cooled to room temperature. Solid K2CO3 (exothermic) was added portionwise until pH = 8-9 was obtained. The mixture was extracted with EtOAc (4x20 mL). The combined organic layers were washed with brine (20 mL), dried (Na2SO4), and concentrated under reduced pressure to give 1.6 g of crude product, which was dried under vacuum for 24 h to give the title compound (850 mg, 81% yield) as an oil, which was used without further purification in the next reaction. 1 H NMR (400MHz, CDCl3) δ6.99(t,J=8.3Hz,1H),6.40(d,J=8.6Hz,1H),2.22~2.06(m,3H).

[0295] Preparation of 4-chloro-3-fluoro-2-methylbenzene-1-sulfonyl chloride (5): Under ice cooling, thionyl chloride (29.1 mL, 395 mmol) was added dropwise to H2O (92.1 mL) over 20 min. The solution containing sulfur dioxide was stirred at 0 °C for 2 h and then at room temperature for 18 h. Separately, HCl (conc.) (23 mL) was added portionwise to compound 4 (3.00 g, 18.8 mmol) at 0 °C to give a beige precipitate. This was stirred at 0 °C for 5 min. A solution of sodium nitrite (1.70 g, 24.4 mmol) in H2O (2 mL) was added dropwise over about 10 min. The above-mentioned sulfur dioxide solution containing copper(I) chloride (38.4 mg, 376 μmol) was slowly added to the reaction mixture over 40 min at 5 °C. The mixture was further stirred under ice cooling for 2 h and then at room temperature for 4 days. The mixture was diluted with CH2Cl2 (20 mL). The aqueous and organic layers were separated. The aqueous layer was extracted with CH2Cl2 (3x20 mL). The combined organic layers were dried (Na2SO4), filtered, and concentrated to give crude compound 5 (1.95 g, 30% yield, 70% purity) as an oil. The crude product was used in the next step without further purification. LCMS: LCMS sample was quenched with N-methylpiperazine (resulting sulfonamide, MW=306.784); (ES + )M+H=307.1; 1 H NMR (400MHz, CDCl3) δ7.81(d,J=8.4Hz,1H),7.46(t,J=7.5Hz,1H),2.69(s,3H).

[0296] 3-Methyl-2-thiophenylsulfonyl chloride:

[0297] [ka]

[0298] Prepared by chlorosulfonylation of 3-methylthiophene as described in US Pat. No. 3,991,081.1 H NMR(CDCl3) δ:7.67(d,J=5.1Hz,1H),7.03(d,J=5.1Hz,1H),2.63(s,3H).

[0299] 3-Chloro-2-thiophenylsulfonyl chloride:

[0300] [ka]

[0301] 3-Chlorothiophene (1.00 g) was dissolved in CHCl3 (10 mL) and the solution was cooled to -30°C. Chlorosulfonic acid (2.4 mL) was added dropwise over 5 min (no noticeable gas evolution). The orange-brown solution was then stirred for 30 min and the temperature was allowed to rise to -10°C and then to room temperature over an additional 30 min. The reaction mixture was then stirred at room temperature for 2 h (no gas evolution was observed and TLC indicated product formation (Rf=0.4 in 8:2 hex / EA)). The reaction mixture was poured onto ice (50 mL), DCM (25 mL) was added and the milky product organic phase was separated, washed with cold water, dried (MgSO4) and concentrated to a yellow oil which was dried under vacuum and used without further purification (0.55 g). 1 H NMR(CDCl3) δ:7.76(d,J=5.7Hz,1H),7.16(d,J=5.7Hz,1H).

[0302] 4-Chloro-3-methylthiophene-2-sulfonyl chloride:

[0303] [ka]

[0304] Preparation of 3-chloro-4-methylthiophene: In a 100 mL flask, copper(I) chloride (5.76 g, 56.5 mmol) was added to 3-bromo-4-methylthiophene (3.16 mL, 28.2 mmol) in DMF (20.1 mL) at room temperature. This was heated at 160 °C in an oil bath for 24 h. The crude product was poured onto H2O (50 mL). The resulting mixture was stirred for 10 min at room temperature. The brown-green solid that formed was filtered and washed with water (3x10.0 mL) and Et2O (4x10.0 mL). The filtrate was extracted with Et2O (3x25.0 mL). The combined organic layers were washed with H2O (2x20.0 mL), brine (20.0 mL), dried (Na2SO4), and concentrated under reduced pressure to give an orange oil (0.890 g, 59% yield of crude product). 1 H NMR (400MHz, CDCl3) δ7.09(d,J=3.5Hz,1H),6.99~6.95(m,1H),2.22~2.21(m,3H).

[0305] Preparation of 4-chloro-3-methylthiophene-2-sulfonyl chloride: 3-Chloro-4-methylthiophene (1.00 g, 7.54 mmol) was dissolved in CHCl3 (4.43 mL) and a solution of chlorosulfonic acid (1.19 mL, 17.3 mmol) in CHCl3 (1.48 mL) was added at room temperature over 5 min. The mixture was stirred for 10 min. Phosphorus pentachloride (4.13 g, 18.9 mmol) was added to the reaction mixture followed by CHCl3 (7.50 mL). It was heated at 50 °C for 1 h. The reaction mixture was slowly added to aq. NaHCO3 + ice (30 mL). It was stirred for 10 min. It was extracted with CH2Cl2 (4x10 mL). The combined organic layers were dried (Na2SO4) and concentrated under reduced pressure to give the title compound (1.30 g, 30% yield, 40% purity) as an oil, which was used without further purification in the next reaction. LCMS: The LCMS sample was quenched with N-methylpiperazine; (ES + )M+H=295.1; 1H NMR (400MHz, CDCl3) δ7.57(s,1H),2.57(s,3H).

[0306] 3,4-Dichlorothiophene-2-sulfonyl chloride:

[0307] [ka]

[0308] Preparation of 3,4-dichlorothiophene: In a 100 mL flask, copper(I) chloride (13.9 g, 137 mmol) was added to 3,4-dibromothiophene (5.03 mL, 45.5 mmol) in DMF (32 mL) at room temperature. This was heated at 160 °C in an oil bath for 24 h. The crude product was poured onto H2O (100 mL) and diluted with Et2O (60 mL). The mixture was stirred for 10 min at room temperature. The brown-green solid that formed was filtered and washed with H2O (3x20 mL) then Et2O (4x20 mL). The filtrate was extracted with Et2O (4x30 mL). The combined organic layers were washed with H2O (2x30 mL), brine (30 mL), dried (MgSO4) and concentrated under reduced pressure to give the title compound 2 (5.50 g, 79% yield) as a red oil. 1 H NMR (400MHz, CDCl3) δ7.21(s,2H).

[0309] Preparation of 3,4-dichlorothiophene-2-sulfonyl chloride: To a solution of compound 2 (1.61 g, 10.5 mmol) in CHCl3 (5.98 mL) was added a solution of chlorosulfonic acid (757 μL, 11.0 mmol) in CHCl3 (1.99 mL) over 5 min. The mixture was stirred at room temperature for 20 min. Phosphorus pentachloride (5.77 g, 26.3 mmol) was added in four portions to the mixture. The mixture was heated at 50 °C for 18 h. The volatiles were removed under reduced pressure and the residue was dissolved in CHCl3 (25 mL) and washed with saturated aqueous NaHCO3 (3x15 mL), H2O (3x10 mL) and brine (10 mL). The organic layer was dried (Na2SO4) and concentrated under reduced pressure to give the title compound 3 (2.32 g, 88% yield). LCMS: LCMS sample was quenched with N-methylpiperazine (resulting sulfonamide, MW=315.240); (ES + )M+H=315.1.

[0310] (3R)-3-Methoxy 1-pyrrolidinesulfonyl chloride:

[0311] [ka]

[0312] Following the procedure described in US Patent Publication US2011 / 0311474A1, (R)-3-methoxypyrrolidine hydrochloride (0.30 g, 2.1 mmol) was suspended in a mixture of 4 mL toluene and 2 mL DCM. Triethylamine (0.64 mL, 4.6 mmol) was added and the mixture was sonicated for 4-5 minutes to give a fine white suspension. In a separate flask, 4 mL toluene was cooled to -40°C in an acetonitrile / dry ice bath. Sulfuryl chloride (0.71 mL, 8.7 mmol) was added and the solution was stirred for 5 minutes. The pyrrolidine suspension was then added dropwise to the cold (-40°C) sulfuryl chloride solution over 10 minutes. The resulting suspension was stirred at the same temperature for 1 hour and then allowed to warm to room temperature. The solids were filtered off and rinsed with toluene. The filtrate was concentrated to give the desired product (0.40 g) as a light brown oil which was used without further purification. 1 H NMR(CDCl3) δ:4.07(tt,J=4.6,2.1Hz,1H),3.48~3.69(m,4H),3.36(s,3H),2.12~2.23(m,1H),1.98~2.12(m,1H).

[0313] (3S)-3-Methoxy 1-pyrrolidinesulfonyl chloride:

[0314] [ka]

[0315] It is prepared in a similar manner to the (R)-isomer, but starting from (S)-3-methoxypyrrolidine hydrochloride.

[0316] Other sulfamoyl chlorides prepared using commercially available secondary amines and in a manner similar to the procedures described in US Patent Application Publication No. US2011 / 0311474A1 include the following:

[0317] (S)-3-Fluoropyrrolidine-1-sulfonyl chloride: Obtained as a white solid from (S)-3-fluoropyrrolidine hydrochloride: 1 H NMR (DMSO-d6) δ:4.54(dt,J=52.6,3.2Hz,1H),2.72~3.03(m,5H),1.34~1.58(m,2H).

[0318] 3,3-Difluoropyrrolidine-1-sulfonyl chloride: Obtained as a white solid from 3,3-difluoropyrrolidine hydrochloride: 1 H NMR(CDCl3) δ:3.81(t,J=12.3Hz,2H),3.74(t,J=7.4Hz,2H),2.53(tt,J=13.0,7.5Hz,2H).

[0319] 3-Methoxyazetidine-1-sulfonyl chloride: Obtained from 3-methoxyazetidine hydrochloride: 1 H NMR (CDCl3) δ:4.22~4.33(m,3H),3.97~4.09(m,2H),3.33(s,3H).

[0320] Preparation of (R)-3-(chloromethyl)pyrrolidine-1-sulfonyl chloride:

[0321] [ka]

[0322] A solution of (R)-3-(hydroxymethyl)pyrrolidine (200 mg, 2 mmol) and triethylamine (0.61 mL, 4.35 mmol) in dry DCM (15 mL) was added dropwise to a stirred solution of sulfuryl chloride (0.48 mL, 5.9 mmol) in DCM (5 mL) at -78 °C. The reaction was stirred at -78 °C for 30 min and then warmed to room temperature for 1 h. The reaction mixture was then washed with 1 M aqueous hydrochloric acid (5 mL) and brine (5 mL). The organic layer was separated, dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound as a colorless oil, which was used as such for the preparation of Example 28.

[0323] Preparation of difluoroaniline hydrochloride intermediate A-5 (Ar=4-methoxyphenyl) from tert-butyl carbamate A-2

[0324] [ka]

[0325] Step 1 - Preparation of aniline intermediate A-3: Nitroarene A-2 (5.00 g, 18 mmol, prepared according to the procedure described in J. Med. Chem. 2003, 46, 1905) and 20% Pd(OH)2 on carbon (130 mg) were suspended in MeOH (50 mL) and the mixture was stirred under a balloon containing hydrogen for 18 h, when TLC analysis (Rf=0.45 in 2:1 hexanes / EtOAc) showed the reduction to be complete. The suspension was removed by filtration through a Celite tube and cooled to 37° C. for 18 h. (登録商標) The mixture was filtered through a pad of ethyl acetate, the catalyst was removed using MeOH to wash, and the solvent was evaporated under reduced pressure. Upon exposure to air, the originally colorless solution turned a very dark greenish-blue color. The crude intermediate aniline A-3 was obtained as a dark greenish-purple foam, which was used immediately in the next step without further purification: 1H NMR (DMSO-d6) δ:8.58(s,1H),6.77(td,J=9.4,2.0Hz,1H),6.60(td,J=9.4,5.5Hz,1H),4.97(s,2H),1.43(s,9H).

[0326] Step 2 - Preparation of sulfonamide A-4 (Ar = 4-methoxyphenyl): The crude aniline A-3 from step 1 (assumed to be 18 mmol) was dissolved in THF (30 mL) and excess 4-methoxyphenylsulfonyl chloride (7.53 g, 36 mmol) was added followed by pyridine (6 mL). The mixture was stirred at 50 °C for 18 h. THF was removed under reduced pressure and the residue was partitioned between EtOAc and water. The extract was washed with saturated aqueous NaHCO3 and brine and dried over MgSO4. The desiccant slurry was passed through a 75 mL pad of silica gel using EtOAc to wash and remove the desiccant and baseline material. Removal of the solvent gave a brown oil which was purified by flash chromatography on silica (~250 mL) using 20-50% EtOAc in hexanes as eluent. After drying under vacuum, the product A-4 was 1 Obtained as a brownish foam (8.16 g) contaminated with unreacted sulfonyl chloride in a 2:1 ratio by H NMR. The material was used directly as is in the next step: 1 H NMR(CDCl3) δ:7.68(d,J=9.0Hz,2H),7.41(td,J=8.8,5.5Hz,1H),6.85~6.98(m,3H),6.57(br.s,1H),5.85(br.s,1H),3.85(s,3H),1.46(s,9H). MS m / z 413.0(MH), m / z 313.0(MH-Boc).

[0327] Step 3 - Preparation of aniline hydrochloride salt A-5 (Ar = 4-methoxyphenyl): The crude carbamate A-4 (8.16 g) from step 3 was stirred in 4N hydrochloric acid in dioxane (25 mL) at room temperature for 1.5 h, during which a beige solid gradually precipitated. After 1.5 h, an additional 10 mL of 4N hydrochloric acid in dioxane was added and stirring was continued for another 1 h. The reaction mixture was then diluted with 50 mL of diethyl ether and the beige precipitate was collected by filtration, washed with ether and dried under vacuum. The aniline salt A-5 (4.38 g) was obtained in pure form in an overall yield of 68% from the nitroarene A-2: 1 H NMR (DMSO-d6) δ:9.73(s,1H),7.62(d,J=8.6Hz,2H),7.06(d,J=9.0Hz,2H),6.69~6.88(m,1H),6.30(td,J=8.6,5.5Hz,1H),3.81(s,3H). MS m / z 313.0(MH).

[0328] Preparation of difluoroaniline hydrochloride intermediate A-5 (Ar=2,3-dichlorophenyl) from acetanilide A-8

[0329] [ka]

[0330] Preparation of acetanilide A-8: Acetanilide A-7 can be prepared by acetylation of 2,6-difluoroaniline A-6 with acetic anhydride following the literature procedure described in Bioorg. Med. Chem. 2016, 24, 2215. Intermediate A-7 is converted to acetanilide A-8 by sequential nitration followed by reduction of the nitro group to the aniline as described in International Publication WO2012 / 101238A1.

[0331] Step 1 - Preparation of sulfonamide A-9 (Ar = 2,3-dichlorophenyl): Aniline A-8 (8.50 g, 45.5 mmol) was dissolved in THF (145 mL) and pyridine (4 equiv, 14.7 mL) was added to the brown solution followed by 2,3-dichlorobenzenesulfonyl chloride (1.2 equiv, 13.45 g). The resulting reaction mixture was stirred at 45 °C for 3.5 h after which the conversion was judged complete as monitored by LCMS. The reaction mixture was cooled to room temperature and then partitioned between EtOAc and 2-Me-THF (1:1) and water. 1N hydrochloric acid solution was added until the pH was slightly acidic. A significant amount of off-white solid was present in the biphasic mixture and was filtered off (1st crop). The layers of the filtrate were separated and the aqueous layer was extracted twice more with EtOAc. The combined organic extracts were washed once with water, then once with brine, dried over MgSO4, filtered, and concentrated down to ~20 mL. The resulting suspension was sonicated, and the solid was collected by filtration and washed with EtOH (2nd crop). Both crops were combined and dried under reduced pressure. A-9 (15.3 g, 85% yield) was obtained as a beige solid that was used without further purification: 1 H NMR(DMSO-d6) δ:10.61(s,1H),9.67(s,1H),7.95(dd,J=8.0,1.4Hz,1H),7.85(dd,J=8.0,1.4Hz,1H),7.51(t,J=8.0Hz,1H),7.05~7.18(m,2H),2.00(s,3H). MS m / z 395.0(MH + ).

[0332] Step 2 - Preparation of aniline hydrochloride A-5 (Ar = 2,3-dichlorophenyl): In a 500 mL round bottom flask, acetanilide A-9 (7.00 g, 17.7 mmol) was suspended in ethanol (65 mL) and a 1:1 mixture of concentrated HCl and water (65 mL) was added. The flask was fitted with a stoppered reflux condenser and heated at 80 °C with stirring. After 24 h, the conversion was -70% as judged by LCMS monitoring. Additional EtOH (65 mL) and 6N hydrochloric acid (65 mL) were added to the suspension and stirring was resumed at 80 °C for an additional 7 h, after which LCMS showed complete conversion to the desired aniline. The reaction mixture was diluted with 50 mL of water while still warm and filtered through a cotton plug to remove a small amount of insoluble material. It was then concentrated to dryness under reduced pressure. The residue was azeotropically dried by evaporating toluene under reduced pressure three times and then dried under vacuum to give 7.2 g of the desired product A-5 as its HCl salt in the form of a yellow solid: 1 H NMR(DMSO-d6) δ:10.30(s,1H),7.93(dd,J=8.2,1.2Hz,1H),7.83(dd,J=8.0,1.4Hz,1H),7.49(t,J=8.0Hz,1H),6.68~6.96(m,1H),6.31(td,J=8.6,5.5Hz,1H). MS m / z 350.9(MH).

[0333] [Table 1] JPEG2024517505000092.jpg216170JPEG2024517505000093.jpg219170JPEG2024517505000094.jpg126170

[0334] General synthetic method A - Preparation of inhibitor I and inhibitor II from intermediate A-5 (Examples 1 and 36):

[0335] [ka]

[0336] Step 1 (Preparation of Example 1 Compound): Chloropyrimidine A-10 (0.546 g, 1.25 equiv.) and aniline hydrochloride A-5 (Ar=4-methoxyphenyl; 0.750 g, 1 equiv.) were dissolved in AcOH (10 mL) and the brown solution was stirred at 50° C. for 1 h. LCMS showed complete conversion to the desired crude product (Example 1). The reaction mixture was cooled to room temperature and diluted with water (50 mL), forming a grey precipitate which was collected by filtration, washed with water, and dried under vacuum. A 100 mg sample of the crude material was purified by passing through a small pad of silica gel (3 mL) using 1:1 hex / EA as the eluent to remove colored baseline material. After removal of the volatiles from the light burgundy solution, the material was lyophilized from MeCN-water to provide the inhibitor of Example 1 (73 mg).

[0337] Step 2 and Step 3 (Preparation of the compound of Example 36): To a solution of the crude thiomethyl derivative (Example 1; 355 mg, 0.72 mmol, 1 equiv.) in 7 mL of DCM, m-CPBA (185 mg, 1.07 mmol, 1.5 equiv.) was added at room temperature. The mixture was stirred at that temperature for 1 h. LCMS shows complete conversion to a 80:20 mixture of sulfoxide and sulfone. The mixture was concentrated to remove most of the DCM, then taken up in EtOAc and washed three times with a solution of NaHCO3. The combined aqueous layers were back-extracted with EtOAc, and the combined organic layers were washed once with water and then with brine. The organic layers were then dried over MgSO4, filtered, and concentrated to give 356 mg of a yellow foam, which was used as is without further purification: MS m / z 507 and 523 (MH + ).

[0338] The crude mixture of sulfoxide and sulfone from above (25 mg, 0.05 mmol) and benzimidazole (12 mg, 0.1 mmol, 2 equiv.) were placed in a 4 mL vial and dissolved in NMP (1 mL). DIEA (43 μL, 0.25 mmol, 5 equiv.) was then added and the resulting mixture was stirred at 60° C. for 19 h (LCMS indicates complete reaction). The reaction was quenched by adding 0.2 mL of AcOH and then diluted to 2 mL with methanol. The solution was filtered and then purified by preparative HPLC (MeOH / H2O / 0.1% HCO2H, gradient from 10% to 100% methanol). Fractions containing the main peak were pooled and partially concentrated to remove methanol. The resulting suspension was dissolved by adding a few milliliters of acetonitrile, and the solution was then frozen and lyophilized to give 8.1 mg of the desired product (Example 36) as a pink solid.

[0339] Other examples of inhibitors of general formula I and general formula II prepared in a similar manner are listed in Tables 1 and 2 below along with characterization data.

[0340] Synthesis of inhibitors I and II using general synthetic method B (examples 12 and 56):

[0341] [ka]

[0342] Step 1: Carbamate A-2 (1.50 g) was dissolved in DCM (5 mL) and TFA (2 mL) was added. After stirring for 2 h at room temperature, the deprotection was complete (LCMS) and the reaction mixture was concentrated and dried under reduced pressure.

[0343] Step 2: The crude TFA salt from step 1 (above) can be used directly in step 2, but the desired intermediate B-2 was contaminated with varying amounts of 8-hydroxy-2-thiomethylpyrimidopyrimidine resulting from the solvolysis of compound A-10. This side reaction was minimized and a cleaner intermediate B-2 was obtained when the aniline TFA salt was neutralized to the free aniline as described below before reacting with compound A-10. The crude TFA salt from step 1 was dissolved in DCM and the solution was washed with NaHCO3. After drying (MgSO4), removal of the volatiles afforded the free aniline B-1 as a brown sticky solid (0.85 g): 1 H NMR (DMSO-d6) δ:7.22~7.38(m,1H),7.06~7.19(m,1H),5.92(brs,2H).

[0344] Chloropyrimidopyrimidine A-10 (550 mg, 2.6 mmol) and B-1 aniline free base (0.39 g, 2.25 mmol) were dissolved in acetic acid (7 mL) and the mixture was stirred at 55° C. for 1 h. LCMS showed complete conversion to the desired product. The reaction mixture was cooled to room temperature and diluted with three volumes of water, resulting in precipitation of the product as a cream colored solid. The material was collected by filtration, washed with water, and dried under vacuum (0.58 g): 1 H NMR (DMSO-d6) δ:10.28(s,1H),9.35(s,1H),8.60(s,1H),8.23~8.49(m,1H),7.58(t,J=8.8Hz,1H),2.71(s,3H). MS m / z 350.1(MH + ).

[0345] Step 3: Nitroarene B-2 (0.86 g, 2.45 mmol) and tin(II) chloride dihydrate (2.7 equiv, 6.6 mmol, 1.49 g) were suspended in absolute ethanol (10 mL) and the mixture was stirred at 65° C. for 3 h. The reaction mixture was partitioned between 1N NaOH and EtOAc. The organic extract was washed with NaHCO3, brine and dried (MgSO4). The desiccant was then separated from the extract by filtration through a pad of silica gel (40 mL) using EtOAc as the eluent to remove baseline material. The filtrate was concentrated and the residue was triturated with EtOAc / hexane to give the aniline B-3 as an orange solid, which was collected by filtration, washed with ether and dried (0.438 g): 1 H NMR (DMSO-d6) δ:9.98(s,1H),9.28(s,1H),8.53(s,1H),6.93(td,J=9.4,1.6Hz,1H),6.77(td,J=9.4,5.5Hz,1H),5.12(brs,2H),2.73(s,3H). MS m / z 321.1(MH + ).

[0346] The mother liquor was purified by flash chromatography (30 mL) using 10% to 70% EtOAc in hexanes (Rf=0.3, 1:2 in hex-EA) to provide an additional 96 mg of aniline B-3.

[0347] Step 4 (Example 12): Aniline B-3 (25 mg, 0.078 mmol) and 4-methoxy-2-methylbenzenesulfonyl chloride (100 mg, 0.23 mmol, 3 equiv.) were dissolved in THF (1 mL) and pyridine (40 μL) was added. The mixture was stirred at 45° C. for 1 h (50% conversion by LCMS). An additional portion of sulfonyl chloride (100 mg) was added and stirring was resumed at 45° C. for 18 h (LCMS shows complete conversion). The reaction mixture was acidified with TFA (100 μL) and diluted to 1.8 mL with DMSO. The product (Example 12) was isolated (13 mg) by preparative HPLC using a gradient of 30% to 100% MeOH + 0.1% HCOOH.

[0348] Step 5 and Step 6 (Example 56): To a suspension of thiomethylpyrimidine (Example 12, 300 mg, 0.59 mmol) in DCM (5 mL) at room temperature was added 1.2 equivalents of m-CPBA (160 mg, 0.71 mmol). The mixture became a yellow solution over 10 min. It was stirred at room temperature for a total of 45 min, at which point LCMS confirmed the reaction was complete. The mixture was concentrated to remove most of the DCM and then partitioned between EtOAc and aqueous NaHCO3. The layers were separated and the organic layer was washed two more times with NaHCO3 solution. The combined aqueous layer was extracted twice with EtOAc and the combined organic layer was washed with brine, then dried over MgSO4 and filtered. The filtrate was concentrated to dryness and then dried under vacuum to give 295 mg of a mixture of sulfoxide and sulfone (~80:20 ratio by LCMS), which was used directly in the next step without further purification.

[0349] 4,5-Dimethyl-1H-imidazole hydrochloride (19 mg, 0.14 mmol, 3 equiv.) and the sulfoxide-sulfone mixture from above (25 mg, 0.048 mmol, 1 equiv.) were placed in a 4 mL vial, followed by NMP (0.5 mL) and DIEA (42 μL, 0.24 mmol, 5 equiv.). The resulting mixture was stirred at 60° C. for 4 h (LCMS shows complete conversion). The reaction mixture was acidified with AcOH (0.2 mL) and the product (Example 56) was isolated by preparative HPLC (MeOH / H2O / 0.1% formic acid, 30% to 100% methanol gradient). Fractions containing the main peak were pooled and partially concentrated to remove methanol. The resulting suspension was dissolved by adding a few milliliters of acetonitrile, and the solution was then frozen and lyophilized. 14 mg of a yellow powder was obtained with 88% purity by HPLC. This material was repurified by flash chromatography on a 3 g silica gel cartridge using a gradient of 100% DCM to 7% isopropanol in DCM. Appropriate fractions were pooled, concentrated, coevaporated once with acetonitrile, and then taken up in a 1:1 MeCN / water mixture. After lyophilization, 7.5 mg of the desired product (Example 56) was obtained as a yellow solid.

[0350] Other examples of inhibitors of general formula I and general formula II prepared in a similar manner are listed in Tables 2 and 3 below along with characterization data.

[0351] Synthesis of inhibitor III and inhibitor IV using general synthetic method C (Examples 29 and 73):

[0352] [ka]

[0353] Step 1: Sulfuryl chloride (0.051 mL, 0.624 mmol) was dissolved in DCM (2 mL) and the solution was cooled to −78° C. A solution of aniline B-3 (50 mg, 0.156 mmol) and triethylamine (0.11 mL, 0.78 mmol) in DCM (5 mL) was added dropwise over 5 min. The reaction mixture was stirred at −78° C. for 90 min to give a solution of intermediate C-1.

[0354] Step 2 (Example 29): (R)-3-methylpyrrolidine hydrochloride (76 mg, 0.62 mmol) in DCM (3 mL) was added to a cold solution of Intermediate C-1. The reaction mixture was allowed to warm to room temperature and then stirred for 2 h (LCMS showed mass of product and reaction completion). The reaction mixture was evaporated to dryness and azeotroped with toluene to remove pyridine. The residue was purified on the ISCO using a RediSep 24 g column (DCM / EtOAc) to provide the inhibitor Example 29 (25 mg) as a brown solid.

[0355] Steps 3 and 4 (Example 73): The thiomethylpyrimidine from step 1 (Example 29, 20 mg, 0.043 mmol) was dissolved in DCM (5 mL) and m-CPBA (11.5 mg, 0.051 mmol) was added. The mixture was stirred at room temperature for 30 min (LCMS showed no more starting material). The reaction mixture was diluted with 25 mL of dichloromethane and washed with saturated NaHCO3 solution. The organic layer was separated and dried over anhydrous Na2SO4 and filtered. The filtrate was evaporated to dryness to provide a mixture of sulfoxide and sulfone (18 mg) as a brown foamy solid, which was used for the next step without any further purification.

[0356] The crude material from above (18 mg, 0.037 mmol) was dissolved in NMP (2 mL) and DIEA (0.033 ml, 0.186 mmol) and benzimidazole (13.2 mg, 0.112 mmol) was added. The reaction mixture was heated at 60° C. for 3 h (LCMS shows complete conversion). The reaction mixture was diluted with MeOH (1 mL), acidified with a few drops of acetic acid and the product was isolated by preparative HPLC using a gradient of 30-100% MeOH-water-0.1% TFA. The product fractions were partially evaporated, dissolved in ACN and water and lyophilized to give Example 73 (12 mg).

[0357] Other examples of inhibitors of general formula I and general formula II prepared in a similar manner are listed in Tables 2 and 3 below along with characterization data.

[0358] Synthesis of inhibitor V using general synthetic method D (Example 88):

[0359] [ka]

[0360] Step 1: Amino-dichloropyrimidopyrimidine D-2 (50 mg, 0.23 mmol, 1 equiv.) and aniline hydrochloride A-5 (85 mg, 0.23 mmol, 1 equiv.) were dissolved in AcOH (1.5 mL) and the mixture was stirred at 55° C. for 1 h (LCMS shows conversion to product, but small amount of aniline remains). An additional 10 mg of dichloro derivative D-2 was added and stirring was continued at 55° C. for 30 min. After cooling to room temperature and dilution with 3 volumes of water, the cream colored precipitate was collected, washed with water, and dried under vacuum (120 mg): 1H NMR(DMSO-d6) δ:10.10(s,1H),9.78(s,1H),8.61(brs,1H),8.39(brs,1H),8.37(s,1H),7.66(d,J=9.0Hz,1H),7.17~7.26 (m,1H),7.13(t,J=9.4Hz,1H),6.94(d,J=2.0Hz,1H),6.86(dd,J=9.0,2.0Hz,1H),3.80(s,3H),2.56(s,3H). MS m / z 508.0(MH + ).

[0361] Step 2: Amino-chloropyrimidine (25 mg, 0.05 mmol, 1 equiv) and benzimidazole (10.5 mg, 0.09 mmol, 1.8 equiv) were suspended in DMSO (0.7 mL) and cesium carbonate (37 mg, 0.11 mmol, 2.3 equiv) was added, followed by copper powder (0.3 mg, 0.1 equiv) and racemic BINOL (1.5 mg, 0.1 equiv). The mixture was stirred at 110° C. for 2 h (LCMS shows complete conversion to the desired mass). The brown reaction mixture was acidified with TFA (100 μL) and the product was isolated by preparative HPLC using a gradient of 30-100% MeOH / 0.1% HCOOH. The inhibitor compound of Example 88 (10 mg) was isolated as a beige solid after lyophilization. The compounds of Examples 85, 86 and 87 were prepared in a similar manner.

[0362] In the case of Example 84, the product was formed using nucleophilic substitution under basic conditions as described in the last step of Scheme A and Scheme B (Intermediate D-3, 4 equivalents of DIEA and 2 equivalents of 3-fluoropyrrolidine hydrochloride were heated at 100° C. for 1.5 hours in NMP).

[0363] Other examples of inhibitors of general formula V prepared in a similar manner are listed in Table 4 below along with characterization data.

[0364] Synthesis of inhibitor VI using general synthetic method E:

[0365] [ka]

[0366] Step 1: Intermediate I (Ar=3-fluoro-2-methylphenyl, Example 90), prepared from the appropriate intermediate A-5, was oxidized to a mixture of sulfoxide and sulfone as described in steps 1 and 2 of general synthetic method A.

[0367] Step 2 and Step 3 (X = CH): Cesium carbonate (1.16 g, 3.51 mmol) and methyl indole-3-carboxylate (X = CH; 0.554 g, 3.1 mmol) were suspended in DMSO (62 mL) and the mixture was stirred at room temperature for 10 min. The mixture of sulfoxide and sulfone from Step 1 (1.55 g, 2.95 mmol) was added and the mixture was stirred at 80 °C for 18 h, at which point conversion was judged complete by LCMS analysis.

[0368] NaOH (236 mg, 5.9 mmol) was added to the reaction mixture from step 2, and the mixture was stirred at room temperature until complete conversion to the desired carboxylic acid intermediate E-1 (X = CH) as determined by LCMS analysis. Then, citric acid solution (1 M) was added to precipitate the product, which was filtered, washed with water, and dried to give compound E-1 (X = CH) (1.67 g, 93% yield) as a brown solid: 1 H NMR(DMSO-d6) δ12.70(br,1H),10.56(s,1H),10.50(s,1H),9.64(s,1H),9.43(s,1H),8.89(d,J=8.3Hz,1H),8.55(s,1H),8.20~8 .12(m,1H),7.61(d,J=7.8Hz,1H),7.53~7.45(m,1H),7.45~7.33(m,3H),7.27(t,J=9.2Hz,1H),2.49~2.48(m,3H). MS m / z 620.3(MH + ).

[0369] Steps 2 and 3 (X=N): Sodium tert-butoxide (341 mg, 3.54 mmol) was added to methyl 1H-indazole-3-carboxylate (573 mg, 3.19 mmol) in dry THF (6.2 mL) and the mixture was stirred at room temperature for 10 min. The sulfoxide / sulfone mixture from step 1 (1.55 g, 2.95 mmol) was added and the mixture was stirred at room temperature for 18 h when conversion was judged complete by LCMS analysis. The reaction mixture was concentrated under reduced pressure to a volume of 20 mL and a solution of NaOH (236 mg, 5.9 mmol) in water (20 mL) was added. The mixture was stirred at room temperature for 4 h when conversion to the desired carboxylic acid was complete by LCMS analysis. The reaction mixture was concentrated under reduced pressure and the residual aqueous solution was acidified with 1 M citric acid to precipitate the product, which was collected by filtration and washed with water to give compound E-1 (X=N) as a brown solid in quantitative yield: 1 H NMR(DMSO-d6) δ10.53(br,1H),10.03(s,1H),9.69(s,1H),8.89(d,J=8.6Hz,1H),8.61( s,1H),8.26(d,J=8.1Hz,1H),7.72~7.67(m,1H),7.62(d,J=7.8Hz,1H),7. 55(dt,J=8.0,1.5Hz,1H),7.48(d,J=8.5Hz,1H),7.41(td,J=8.0,5.6Hz,1 H),7.33(td,J=8.6,5.9Hz,1H),7.25(t,J=9.3Hz,1H),2.50~2.49(m,3H). MS m / z 607.2(MH + ).

[0370] Step 4 (Preparation of the compound of Example 103, X = CH): Carboxylic acid E-1 (X = CH; 61 mg, 0.1 mmol) and (S)-3-hydroxypiperidine hydrochloride (17 mg, 0.12 mmol) were dissolved in DMF (0.5 mL) and DIPEA (61 μL, 0.35 mmol) was added, followed by HATU (58 mg, 0.15 mmol). The mixture was stirred for 18 h at ambient temperature. The reaction mixture was then directly purified by preparative reverse-phase HPLC to provide the compound of Example 103 as a yellow powder after lyophilization.

[0371] Other example compounds (X = CH or N) listed in Table 3 and using Method E were prepared in a similar manner.

[0372] Synthesis of inhibitor II using general synthetic method F (Example 95):

[0373] [ka]

[0374] Step 1: 3-Indole sulfonyl chloride was prepared as described in Org. Lett. 2011, 13, 3588. The sulfonyl chloride (200 mg, 0.9 mmol) was placed in a 25 mL flask, to which THF (4 mL) was added, followed by dimethylamine hydrochloride (2 eq., 150 mg, 1.9 mmol) and DIEA (4 eq., 0.65 mL, 3.7 mmol). The solution immediately turned pale yellow and a yellow gummy oil was deposited on the bottom. Stirred at room temperature for 20 min (LCMS showed complete consumption of the sulfonyl chloride). The mixture was partitioned between EtOAc and a saturated solution of NH4Cl. The aqueous layer was extracted with EtOAc, and the combined organic layers were washed once more with a saturated solution of NH4Cl, then with brine. It was then dried over MgSO4, filtered, and concentrated to dryness to give 65 mg of a beige crystalline solid, which was used as is without further purification: 1H NMR(DMSO-d6) δ:12.17(brs,1H),7.96(d,J=3.1Hz,1H),7.81(d,J=7.8Hz,1H),7.49~7.57(m,1H),7.22~7.28(m,1H),7.16~7.22(m,1H),2.58(s,6H). MS m / z 225.1(MH + ).

[0375] Step 2: The thiomethyl derivative I (Ar = 3-fluoro-2-methylphenyl; Example 90) was oxidized to a mixture of the sulfoxide and sulfone as described in general method A.

[0376] Step 3 (example 95): Prepared using general method A using the indole sulfonamide from step 1 and the sulfoxide / sulfone mixture from step 2.

[0377] Other example compounds listed in Table 3 were prepared in a similar manner using general method F.

[0378] Synthesis of inhibitor VIII using general synthetic method G (Example 142):

[0379] [ka]

[0380] Step 1: To a solution of 1H-indol-3-yl-thiocyanate (Phosphorus, Sulfur and Silicon and the Related Elements 2014, 189, 1378) (100 mg, 0.57 mmol) in iPrOH (15 mL) was added sodium sulfide anhydrate (414 mg, 1.72 mmol) dissolved in 0.5 mL of water, and the resulting mixture was then stirred at 50° C. for 2 hours. After this, 4-chlorotetrahydropyran (0.19 mL, 1.72 mmol) was added and stirred at 50° C. overnight. The reaction mixture was diluted with EtOAc (30 mL) and separated. The organic layer was washed with water (15 mL), followed by brine (15 mL), dried over MgSO4, and then concentrated in vacuo to give the crude sulfide, which was used directly in the next step without further purification.

[0381] Step 2: The sulfide from step 1 was dissolved in DCM and 3-chloroperoxybenzoic acid (297 mg, 1.72 mmol) was added and stirred at room temperature for 2 h. After completion, the reaction was quenched by adding 10 mL of a 1:1 solution of saturated aqueous NaHCO3 and 10% aqueous Na2SO3. The resulting suspension was stirred at room temperature for 15 min. EtOAc was added and the organic layer was separated. The organic layer was washed with water (15 mL) and then with saturated aqueous brine solution (15 mL). The organic layer was separated, dried (MgSO4), and concentrated to dryness after filtration to give the expected sulfone (154 mg, 98%), which was dissolved in DMSO and used directly in the next step without further purification. MS m / z 266.2 (MH + ).

[0382] Step 3 (Example 142): The indole from step 2 was coupled to intermediate I (Ar=2,3-dichlorophenyl) using the procedure described in general method A.

[0383] Examples of other inhibitors prepared in a similar manner using an appropriate alkylating agent in step 1 are listed under Method F in Table 3.

[0384] Synthesis of inhibitor IX using general synthetic method H (Example 130):

[0385] Using iron as a reducing agent in step 2 (Example 130):

[0386] [ka]

[0387] Step 1: 4-Methylpiperidin-4-ol (0.24 g, 1.84 mmol) and potassium carbonate (0.49 g, 3.52 mmol) were added to a solution of 3-fluoro-2-nitro-aniline (0.25 g, 1.60 mmol) in MeCN (2.6 mL). The resulting mixture was stirred at 85° C. for 10 h. MeCN was removed under reduced pressure and EtOAc was added. The suspension was centrifuged and poured into a flask. The solution was concentrated and the 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 + ).

[0388] Step 2 (using iron as reducing agent): 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) and formic acid (1.9 mL, 49.6 mmol) in iPrOH (6.5 mL). The resulting mixture was heated to 90° C. and stirred for 10 h. The reaction mixture was cooled to room temperature and then filtered through Celite. (登録商標)The solution was concentrated and the crude product was purified by column chromatography (silica gel, 0-15% MeOH in DCM) to give 1-(1H-benzimidazol-4-yl)-4-methyl-piperidin-4-ol (0.17 g, 55% yield) as a reddish foamy solid. MS m / z 232.2 (MH + ). 1 H NMR(400MHz,DMSO-d6) d:12.21(brs,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).

[0389] Step 3 (Example 130): The benzimidazole from step 2 was coupled to intermediate I (Ar=2,3-dichlorophenyl) using the procedure described in general method A.

[0390] Use of zinc as reducing agent in step 2 (preparation of benzimidazole fragment C328):

[0391] Step 1: 4-Methylpiperidin-4-ol was replaced by (R)-3-methoxypiperidine hydrochloride. The nitroarene from step 1 was obtained as a red solid in 88% yield and was used directly in step 2 without purification: MS m / z 252.1 (MH + ). 1H NMR(400MHz, CDCl3) d:1.2~1.35(m,1H),1.58~1.73(m,1H),1.74~1.85(m,1H),2.06~2.17(m ,1H),2.53~2.61(m,1H),2.70(td,J=11.5,3.0Hz,1H),3.12(dt,J=12.0, 3.8Hz,1H),3.35~3.45(m,2H),3.40(s,3H),4.79(brs,2H),6.38(dd,J=8.25,1.13Hz,1H),6.41(dd,J=8.13,1.13Hz,1H),7.12(t,J=8.13Hz,1H).

[0392] Process 2: Teflon (登録商標) In a 250 mL round bottom flask equipped with a coated magnetic stir bar, under a nitrogen atmosphere, a mixture of the crude product from step 1 (1.37 g, 5.45 mmol) and ammonium chloride (4.08 g, 76.3 mmol) in methanol (18 mL) and 2-methyltetrahydrofuran (36 mL) was treated with zinc dust (2.7 g, 38.16 mmol) added in one portion. An exotherm was observed and the reaction mixture became colorless after .about.10 min. The reaction mixture was stirred for 1 h and LCMS showed clean conversion to 1,2-diaminobenzene. The reaction mixture was filtered through a small celite pad, which was washed with DCM:isopropanol (9:1, 30 mL). The filtrate was washed with DCM-isopropanol (9:1, 100 mL) and then with 10% aqueous potassium carbonate (30 mL, pH 10). The organic phase was collected and dried over anhydrous magnesium sulfate. Evaporation of the solvent under reduced pressure gave 1.2 g: MS m / z 222.2 (MH + ). 1H NMR(400MHz, CDCl3) d:1.4~1.6(brs,1H),1.61~1.74(m,1H),1.81~1.94(m,1H),2.0(brs,1H),2.65(brs,1H),2.95(brs,1H),3.17(brs,1H),3.37(brs,2 H),3.4~3.5(m,2H),3.41(s,3H),3.79(brs,2H),6.53(dd,J=7.63,1.45Hz,1H),6.61(dd,J=7.94,1.45Hz,1H),6.68(t,J=7.8Hz,1H).

[0393] The crude 1,2-dianiline was cyclized to the desired benzimidazole as follows: (登録商標) A 100 mL round bottom flask under nitrogen equipped with a coated magnetic stir bar and reflux condenser was charged with 2-propanol (20 mL) and the crude product from above (1.20 g, 5.42 mmol). Formic acid (5 mL, 132 mmol) was then added in one portion and the resulting solution was heated at 60° C. for 16 h. LCMS showed clean formation of the desired benzimidazole (m / z 232). The cooled reaction mixture was diluted with DCM:2-propanol (9:1, 200 mL), washed with 10% aqueous potassium carbonate (40 mL, pH 10) and brine, and dried over anhydrous magnesium sulfate. Evaporation of the solvent under reduced pressure gave a brown solid. The solid was chromatographed on silica gel (3 x 12 cm) and eluted with ethyl acetate-2-propanol (90:5 to 9:1) to give 1.01 g (81% yield). Trituration with ethyl acetate (10 mL) gave 0.88 g of a pale orange-brown solid: MS m / z 232.1 (MH + ). 1H NMR (400 MHz, CDCl3) δ: 1.47 (brs, 1H), 1.80 (br.s, 1H), 1.94 (br.s, 1H), 2.13 (br.s, 1H), 2.92 (br.s, 2H), 3.47 (s, 3H), 3.53-3.63 (m, 1H), 4.1 (br.s, 2H), 6.72 (br.s, 1H), 7.07 (br.s, 1H), 7.19 (t, J = 7.9 Hz, 1H), 7.98 (s, 1H), 9.43 and 9.7 (two br.s, 2:1 ratio, 1H).

[0394] Other examples of inhibitors prepared in a similar manner, using the appropriate amine in step 1 and either iron or zinc as the reducing agent in step 2, are listed under Method H in Table 3 below.

[0395] Synthesis of inhibitor X using general synthetic method I (Example 138):

[0396] [ka]

[0397] Step 1: 1-Oxa-8-azaspiro[4.5]decane hydrochloride (188 mg, 1.06 mmol) and potassium carbonate (2.20 equiv, 292 mg, 2.11 mmol) were added to a deep red solution of 3-fluoro-2-nitro-aniline (150 mg, 0.961 mmol) in ACN (4.5 mL). The resulting mixture was stirred at 90° C. for 16 h. Upon completion, the reaction mixture was diluted with ACN and centrifuged. The supernatant was separated and used as is in the next step. MS m / z 278.3 (MH + ).

[0398] Step 2: To 2-nitro-3-(1-oxa-8-azaspiro[4.5]decan-8-yl)aniline (250 mg, 0.901 mmol) in ACN was added ammonium chloride (1028 mg, 19.2 mmol) and zinc (628 mg, 9.61 mmol). The resulting suspension was stirred at 40° C. for 1 h. Upon completion, the reaction mixture was diluted with EtOAc and then centrifuged. The supernatant was separated and centrifuged under vacuum. The residue, 3-(1-oxa-8-azaspiro[4.5]decan-8-yl)benzene-1,2-diamine (194 mg, 0.784 mmol, 82% yield), was used as is in the next step. MS m / z 248.2 (MH + ).

[0399] Step 3: 3-(1-oxa-8-azaspiro[4.5]decan-8-yl)benzene-1,2-diamine (194 mg, 0.78 mmol) was dissolved in AcOH (6 mL), then sodium nitrite (54 mg, 0.78 mmol) was added and stirred for 1 h at room temperature. After completion, EtOAc and NaHCO3 aqueous were added and the organic layer was separated. The organic layer was washed with NaHCO3 aqueous, then with brine, dried over MgSO4, filtered and concentrated under vacuum to give 8-(1H-benzotriazol-4-yl)-1-oxa-8-azaspiro[4.5]decane (165 mg, 0.64 mmol, 67% yield), which was used without purification. MS m / z 259.2 (MH + ).

[0400] Step 4 (Example 138): The benzotriazole from step 3 was coupled to intermediate I (Ar=2,3-dichlorophenyl) using the procedure described in general method A.

[0401] Other examples of inhibitors prepared in a similar manner using the appropriate amine in step 1 are listed under Method I in Table 3.

[0402] Preparation of the compound of Example 114:

[0403] [ka]

[0404] Step 1 - Preparation of 4-(pyridin-3-yl)-1H-benzo[d]imidazole: Bromobenzimidazole (70 mg, 0.355 mmol), potassium carbonate (196 mg, 1.42 mmol) and 3-pyridylboronic acid (57 mg, 0.46 mmol) were placed in a 4 mL vial and dioxane (2 mL) and water (0.7 mL) were added. Argon gas was bubbled through the mixture for 1 min and then tetrakis(triphenylphosphine)palladium(0) (16.4 mg, 0.014 mmol) was added. Argon gas was bubbled through the solution for 3 min, the vial was sealed and heated at 100° C. for 2 h (conversion to the desired product was complete as judged by LCMS analysis). The reaction mixture was cooled to room temperature, diluted with EtOAc and washed with brine. After drying over MgSO4, the extract was concentrated under reduced pressure and the residue was purified by flash chromatography using Et3N pre-treated silica and a DCM-20% iPrOH / DCM gradient to provide the desired benzimidazole intermediate (58 mg, 84% yield): 1 H NMR(DMSO-d6) d:12.71(bs,1H),9.24(s.1H),8.57(dd,J=5.1,1.6Hz,1H),8.43(bd,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 + ).

[0405] Step 2: The thiomethyl derivative I (Ar=3-fluoro-2-methylphenyl; Example 90) was oxidized to a mixture of the sulfoxide and sulfone as described in general procedure A.

[0406] Step 3 (Example 114): Prepared using general method A from the benzimidazole derivative described in step 1 and the sulfoxide / sulfone mixture described in step 2.

[0407] Preparation of (R)-3-(difluoromethoxy)pyrrolidine hydrochloride:

[0408] [ka]

[0409] (R)-N-Boc-3-hydroxypyrrolidine was difluoromethylated using 2-fluorosulfonyl-2,2-difluoroacetic acid and copper(I) iodide catalyst as described in J. Org. Chem. 2016, 81, 5803, and the N-Boc protecting group was subsequently removed with 4N hydrochloric acid in dioxane.

[0410] Preparation of (S)-3-ethylpyrrolidine hydrochloride:

[0411] [ka]

[0412] Step 1: Commercially available (R)-2-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl)acetic acid (2.0 g, 8.72 mmol) was dissolved in dry THF (25 mL) and 1 M BH3 in THF (17.45 mL, 17.45 mmol) was added. The reaction mixture was stirred at room temperature for 3 h. It was then cooled in an ice-water bath and quenched by slowly adding 1 N hydrochloric acid. The product was extracted into EtOAc and washed with saturated aqueous NaHCO3 and brine. The organic layer was dried over anhydrous Na2SO4 and evaporated to dryness to provide (R)-tert-butyl 3-(2-hydroxyethyl)pyrrolidine-1-carboxylate (1.50 g, 6.98 mmol, 80% yield) as an oil:1 H NMR(CDCl3) δ:3.62~3.70(m,2H),3.37~3.60(m,2H),3.15~3.33(m,1H),2.88(dt,J=18.4,9.6Hz,1H),2.13~2.35 (m,1H),1.94~2.08(m,1H),1.47~1.69(m,3H),1.45(s,9H),1.30~1.43(m,1H),0.93(t,J=7.4Hz,1H).

[0413] Step 2: (R)-tert-Butyl 3-(2-hydroxyethyl)pyrrolidine-1-carboxylate (1.0 g, 4.64 mmol) from step 1 was dissolved in DCM (15 mL) and triethylamine (1.55 mL, 11 mmol) was added. The mixture was cooled to 0° C. and a solution of methanesulfonyl chloride (0.58 mL, 7.4 mmol) in DCM (2 mL) was added dropwise. The reaction mixture was stirred at 0° C. for about 1 h. The reaction mixture was then diluted with DCM (15 mL) and washed with saturated NaHCO3. The organic layer was separated, dried over anhydrous Na2SO4, and filtered. The filtrate was evaporated to dryness to give the crude mesylate (1.36 g) as an oil, which was used in the next step without further purification: 1 H NMR(CDCl3) δ:4.26(dt,J=11.3,6.3Hz,2H),3.39~3.66(m,2H),3.21~3.35(m,J=8.2,8.2Hz,1H),3.03(s,3H),2.85~2.99(m,1H),2.21~2 .38(m,1H),1.98~2.15(m,1H),1.80~1.92(m,1H),1.70~1.80(m,1H),1.48~1.70(m,2H),1.46(s,9H),0.97(t,J=7.2Hz,1H).

[0414] Step 3: The crude mesylate from step 2 (0.60 g, 2.0 mmol) was dissolved in THF (10 mL) and the solution was cooled in an ice-water bath. 1 M lithium triethylborohydride in THF (4.70 mL, 4.70 mmol) was slowly added, then the ice bath was removed and the mixture was stirred at room temperature for 2 h. TLC (2:1, ethyl acetate / hexanes) showed no starting material. Methanol (5 mL) was slowly added to quench the reaction and the organic solvent was removed under reduced pressure. The crude reaction mixture was partitioned between EtOAc (30 mL) and water (15 mL) and the organic layer was washed with brine (10 mL). The organic layer was separated, dried over anhydrous Na2SO4, filtered and the filtrate was evaporated to dryness to provide the crude product. The material was purified on the ISCO using a RediSep 12 g column (Hex / EtOAc) to give (S)-tert-butyl 3-ethylpyrrolidine-1-carboxylate (300 mg, 73% yield) as an oil: 1 H NMR(CDCl3) δ:3.48(tt,J=29.7,9.4Hz,2H),3.16~3.34(m,1H),2.85(dt,J=19.8,10.1Hz, 1H), 1.90~2.11 (m, 2H), 1.45 (s, 9H), 1.36~1.43 (m, 3H), 0.93 (t, J=7.4Hz, 3H).

[0415] Step 4: The carboxylate from step 3 (250 mg, 1.25 mmol) was dissolved in MeOH (2 mL) and 4 M hydrochloric acid in dioxane (2 mL, 8 mmol) was added. The reaction was stirred at room temperature for 2 h. The volatiles were then removed under reduced pressure and the residue was dried under vacuum to provide (S)-3-ethylpyrrolidine hydrochloride (162 mg, 95% yield) as a thick oil.

[0416] Preparation of (R)-3-(methoxymethyl)pyrrolidine hydrochloride:

[0417] [ka]

[0418] Step 1: (R)-tert-Butyl 3-(hydroxymethyl)pyrrolidine-1-carboxylate (1.00 g, 4.97 mmol) was dissolved in THF (20 mL) and the solution was cooled in an ice-water bath. 60% NaH oil dispersion (0.60 g, 14.91 mmol) was added portionwise and the reaction mixture was stirred at 0° C. for 15 min. Iodomethane (1.55 mL, 24 mmol) was added slowly and the reaction mixture was stirred at room temperature overnight. The reaction mixture was then quenched with saturated NH4Cl solution and extracted with EtOAc (2×50 mL). The organic layer was separated, dried over anhydrous Na2SO4, and filtered. The filtrate was evaporated to dryness and purified by silica gel column chromatography (80 g) using 0-100% EtOAc-hex. (R)-tert-Butyl 3-(methoxymethyl)pyrrolidine-1-carboxylate (700 mg, 3.25 mmol, 65.4% yield) was obtained as a colorless oil: 1 H NMR(CDCl3) δ:3.49(dd,J=11.0,7.6Hz,1H),3.43(brs,1H),3.35(s,3H),3.26~3.35(m,3H),3.06(dd,J=1 0.7,7.3Hz,1H),2.46(spt,J=7.3Hz,1H),1.91~2.02(m,1H),1.58~1.70(m,1H),1.46(s,9H).

[0419] Step 2: (R)-tert-butyl 3-(methoxymethyl)pyrrolidine-1-carboxylate (100 mg, 0.46 mmol) from step 1 in DCM (2 mL) was mixed with 4 M hydrochloric acid in dioxane (2 mL, 8.0 mmol) and the mixture was stirred at room temperature for 2 h. The volatiles were evaporated under reduced pressure, the residue was coevaporated with EtOAc to dryness and the product was dried under vacuum to provide (R)-3-(methoxymethyl)pyrrolidine hydrochloride (70 mg) as an oil, which was used without further purification.

[0420] Preparation of the compound of Example 89:

[0421] [ka]

[0422] Step 1: 2-Chloro-6-fluoroaniline (10 g) was placed in a 250 mL flask and dissolved in 40 mL of glacial acetic acid. Acetic anhydride (7.47 mL) was added at room temperature and the resulting mixture was stirred at 90° C. for 1 h, at which point LCMS analysis confirmed that the reaction was complete. The volatiles were removed under reduced pressure, the residue was dissolved in DCM and slowly neutralized with a saturated solution of NaHCO3. The layers were separated and the aqueous layer was extracted three times with DCM. The combined organic layers were washed once with water, then dried over MgSO4, filtered and concentrated. After drying in vacuum, the desired product was obtained as white to pale pink crystals (12.79 g): 1 H NMR (CDCl3) δ:7.16~7.26(m,2H),7.03~7.13(m,1H),6.93(brs,1H),2.23(brs,3H). MS m / z 188.1(MH + ).

[0423] Step 2: The acetanilide from step 1 (12.75 g) was taken up in 25 mL of concentrated sulfuric acid and cooled to 0° C. in an ice bath. Nitric acid (90%, 3.31 mL) was added slowly. After 5-10 min, the mixture became a solid mass. It was allowed to warm to room temperature, which produced a thick burgundy sludge. After a total of 4 h, the reaction was monitored by LCMS, which confirmed that some starting material remained. An additional 5 mL of sulfuric acid was added to improve fluidity, followed by 0.3 mL of 90% nitric acid. The mixture was stirred at room temperature for an additional 18 h. The mixture was then cooled to 0° C. and poured onto crushed ice (~150 mL). Once the ice had melted, the suspension was sonicated and the yellow solid was collected by filtration, washed with water, and dried (15.1 g crude product). The crude solid was taken up in 50 mL of acetonitrile and brought to reflux to give a clear dark red solution. The heat was turned off and the mixture was allowed to cool to room temperature over 1 hour, then stirred at room temperature for 2 hours. By that time the mixture had turned into a solid mass, which was broken up with a spatula and sonicated. The solid was collected by filtration and washed with a small amount of cold acetonitrile. The desired off-white nitro compound (6.63 g) was obtained as a single regioisomer, as shown by NMR (the mother liquor gave a second crop of 2.16 g containing 7% of 6-chloro-2-fluoro-3-nitroacetanilide): 1 H NMR (CDCl3) δ:7.90 (dd,J=9.2,4.9Hz,1H),7.24(dd,J=9.2,8.4Hz,1H),6.98(brs,1H),2.28(s,3H). MS m / z 233.0(MH + ).

[0424] Step 3: To a solution of the nitroacetanilide from step 2 (500 mg, 2.15 mmol) in 15 mL of ethanol was added a solution of NH4Cl (60 mg, 1.12 mmol) in 1.35 mL of water. The mixture was warmed to 70° C., and then iron powder (600 mg, 10.75 mmol) was added in three portions at 10 min intervals. The resulting dark red to burgundy colored mixture was stirred at 70° C. for 20 h, at which point LCMS of a filtered aliquot of the reaction mixture confirmed the reaction was complete. The mixture was filtered through celite and concentrated to 100° C. for 20 h. (登録商標) The dark brown filtrate was concentrated to dryness and then taken up in EtOAc, to which MgSO4 was added. The suspension was stirred and then filtered to give a clear pale yellow solution. The solution was concentrated to dryness to give the desired product (440 mg) as a pale yellow solid, which was used as is without further purification: 1 H NMR(CDCl3) δ:6.92 (t,J=9.0Hz,1H),6.76(brs,1H),6.68(dd,J=8.6,4.7Hz,1H),3.98(brs,2H),2.24(brs,3H). MS m / z 203.1(MH + ).

[0425] Step 4: The aniline from step 3 was sulfonylated with 4-methoxybenzenesulfonyl chloride in the general manner described for A-9 in Scheme A: 1 H NMR(DMSO-d6) δ:9.89(s,1H),9.67(s,1H),7.57~7.74(m,2H),7.23(t,J=9.2Hz,1H),7 .13(dd,J=8.8,5.3Hz,1H),7.02~7.10(m,2H),3.82(s,3H),2.00(s,3H). MS m / z 373.0(MH + ).

[0426] Step 5: The acetanilide from step 4 (200 mg, 0.54 mmol) was taken up in 1.5 mL of ethanol, then a 1:1 mixture of concentrated HCl and water (2 mL) was slowly added. The yellow slurry was then brought to 80° C. and stirred for 1 h. At that point, 1 mL of ethanol was added to improve solubility. The mixture was stirred at the same temperature for an additional 5 h (at which point the mixture had become a clear yellow solution), at which point LCMS analysis indicated less than 3% starting material remaining. The mixture was concentrated to remove most of the ethanol and then chilled on ice. It was basified with 4N NaOH to pH 5-6. The resulting suspension was sonicated, and the solid was collected by filtration and washed with water. After drying under reduced pressure, 156 mg of the desired product was obtained as a beige solid: 1 H NMR(DMSO-d6) δ:9.53(s,1H),7.53~7.72(m,2H),7.00~7.14(m,2H),6.95(dd,J=10.8,8.8Hz,1H),6.36(dd,J=8.6,5.1Hz,1H),5.39(s,2H),3.81(s,3H). MS m / z 329.0(MH).

[0427] Step 6: Using the protocol described in Scheme A for preparing inhibitors of Formula I from synthetic intermediate A-5 (Step 1, Example 1), the aniline from Step 5 was reacted with intermediate A-10 to give the compound of Example 89.

[0428] Preparation of the compound of Example 135:

[0429] [ka]

[0430] Step 1: To a 100 mL round bottom flask was added 2,6-difluoronitrobenzene (1.3 mL, 12.6 mmol) and ethyl cyanoacetate (1.6 mL, 15.1 mmol) in DMF (15 mL). Sodium hydride (754 mg, 18.9 mmol) was then added slowly at room temperature. The reaction was stirred at room temperature for 15 minutes. The reaction was quenched with 1 M hydrochloric acid until the deep red solution turned yellow, and then diluted with EtOAc. The organic layer was separated and then washed with aqueous NH4Cl, followed by brine. The organic layer was dried over MgSO4, filtered, and then concentrated under reduced pressure. The crude material was dissolved in DMSO (9 mL) and water (1 mL) and transferred to a 20 mL microwave vial. The reaction was heated to 120 °C and stirred for 16 h. The reaction mixture was cooled to room temperature and diluted with EtOAc, then washed with aqueous NH4Cl, followed by brine. The organic layer was dried over MgSO4, filtered, and then concentrated under reduced pressure. The crude material was purified by normal phase flash column chromatography using hexanes:EtOAc to give the desired benzyl nitrile (2.12 g, 93%) as an orange solid. 1 H NMR (400MHz, DMSO-d6) δ:7.80(td,J=7.8,5.5Hz,1H),7.65(t,J=9.6Hz,1H),7.54(d,J=7.4Hz,1H),4.28(s,2H). MS m / z 725.4(MH + ). MS m / z 181.2(MH + ).

[0431] Step 2: The phenylacetonitrile derivative from step 1 (200 mg, 1.11 mmol) and DMSO (4 mL) were placed in a 25 mL flask. Then, diphenyl(vinyl)sulfonium-trifluoromethanesulfonate (479 mg, 1.32 mmol) was added, followed by 1,8-diazabicyclo[5.4.0]undec-7-ene (0.37 ml, 2.48 mmol) at room temperature. The mixture was stirred at that temperature for 16 h. After completion, an aqueous solution of NH4Cl was added, and the aqueous layer was extracted with EtOAc. The organic layer was washed with water and once with brine. The organic layer was dried over MgSO4, filtered, and concentrated under vacuum. The resulting residue was purified by flash chromatography through a silica gel column using EtOAc in hexane to give the desired cyclopropane derivative (138 mg, 60% yield). 1 H NMR (400MHz, CDCl3) δ:7.47~7.59(m,1H),7.29~7.41(m,2H),1.69~1.85(m,2H),1.29~1.39(m,2H). MS m / z 207.2(MH + ).

[0432] Step 3: To a 10 mL microwave vial was added the cyclopropane derivative from step 2 (138 mg, 0.67 mmol). Then, 3 mL of concentrated ammonium hydroxide was added to the reaction. The reaction was heated in a microwave at 130° C. for 1 h. After completion, the reaction was diluted with water and then extracted with EtOAc. The organic layer was washed with brine, dried over MgSO4, filtered, and then concentrated under reduced pressure to give 1-(3-amino-2-nitro-phenyl)cyclopropanecarbonitrile (120 mg, 88% yield) as an orange solid. 1 H NMR (400MHz, CDCl3) δ:7.27(d,J=15.26Hz,1H),6.83(d,J=8.38Hz,1H),6.86(d,J=7.50Hz,1H),5.36(brs,2H),1.71(brs,2H),1.24(brs,2H).

[0433] Step 4: Reduction and ring closure of the intermediate 1,2-phenylenediamine to the benzotriazole ring was accomplished using the procedures described in steps 2 and 3 of general method I for example 138.

[0434] Step 5 (Example 135): The benzotriazole from step 4 was coupled to intermediate I (Ar=2,3-dichlorophenyl) using the procedure described in general method A.

[0435] Preparation of the compound of Example 137:

[0436] [ka]

[0437] Step 1: 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 h. Water was added and the aqueous mixture was extracted with EtOAc. The organic layers were combined, washed with brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by column chromatography (silica gel, 0-100% EtOAc in hexane) to give 3-(2-methoxyethoxy)-2-nitro-aniline (52 mg, 38% yield). MS m / z 213.1 (MH + ).

[0438] Step 2: Reduction and ring closure of the intermediate 1,2-phenylenediamine to the benzotriazole ring was accomplished using the procedures described in steps 2 and 3 of general method I for example 138.

[0439] Step 3 (Example 137): The benzenetriazole from step 2 was coupled to intermediate I (Ar=2,3-dichlorophenyl) using the procedure described in general method A.

[0440] Preparation of compounds of Examples 160-163 (Table 5):

[0441] [ka]

[0442] Step 1: Carbomethoxylation of 2,4,5-trifluoroaniline was carried out as described in patent WO2020 / 261156.

[0443] 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 at 70° C. for 16 h. The reaction was monitored by LCMS. When the reaction was complete, it was quenched with 1 M hydrochloric acid. The aqueous layer was extracted three times with EtOAc (15 mL). The combined organic layers were washed with brine, dried over MgSO4, and evaporated to dryness. The residue was purified by chromatography on a silica gel column using 0-30% EtOAc in hexanes. Pure fractions were collected and evaporated to give methyl 3-((2,3-dichlorophenyl)sulfonamido)-2,5,6-trifluorobenzoate (5.14 g, 91% yield) as a light brown solid: m / z=412.0.

[0444] Step 3: To a solution of methyl 3-((2,3-dichlorophenyl)sulfonamido)-2,5,6-trifluorobenzoate (5.14 g, 12.4 mmol) from step 2 in 30 mL of THF:MeOH (5:1) was added 2 M KOH (37 mL, 74.5 mmol) at room temperature. The reaction was stirred at room temperature overnight. When the reaction was complete, it was evaporated to dryness, and to the residue was added water (30 mL) and diethyl ether (30 mL). The aqueous layer was washed twice with ether (20 mL). The aqueous layer was acidified with 1 M hydrochloric acid to pH=2. The aqueous layer was extracted three times with EtOAc (30 mL). The combined organic layers were washed with brine, dried over MgSO4, filtered, and evaporated to give 3-((2,3-dichlorophenyl)sulfonamido)-2,5,6-trifluorobenzoic acid (4.5 g, 91% yield) as a pale orange oil. The compound was used directly in the next step: m / z=398.0.

[0445] Step 4: To a solution of 3-((2,3-dichlorophenyl)sulfonamido)-2,5,6-trifluorobenzoic acid (4.50 g, 11.2 mmol) from step 3 in acetonitrile (30 mL) was added triethylamine (1.71 mL, 12.37 mmol) and diphenylphosphoryl azide (2.91 mL, 13.50 mmol) at room temperature. The reaction was heated at 80° C. overnight. The reaction was cooled to room temperature and water (30 mL) was added. The aqueous layer was extracted three times with EtOAc (30 mL). The combined organic layers were washed with brine, dried over MgSO4, 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: m / z=391.0.

[0446] 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 4M LiOH aqueous (17 mL). The pressure vessel was screwed and heated in an oil bath at 100° C. for 1 h. When the reaction was complete, it was quenched with saturated NH4Cl, EtOAc was added, and the layers were separated. The aqueous layer was extracted twice with EtOAc (20 mL). The combined organic layers were washed with brine, dried over MgSO4, filtered, and evaporated to give N-(3-amino-2,4,5-trifluorophenyl)-2,3-dichlorobenzenesulfonamide (1.09 g, 86% yield) as a brown solid. The compound was carried on to the next step without further purification: 1 H NMR(400MHz,DMSO-d6) δ:10.59(br.s.,1H),7.94(dd,J=8.2,1.6Hz,1H),7.89(dd,J=8.2,1.6Hz,1H),7.51(dd,J=8.0Hz,1H),6.34~6.43(m,1H),5.72(s,2H). m / z=369.0.

[0447] Step 6: The aniline from step 5 was converted to the pyrimidopyrimidine where Ar = 2,6-dichlorophenyl, as described under general method A (example 36).

[0448] Step 7 (Examples 160-163, Table 5): The thiomethyl intermediate from step 6 was converted to the inhibitor using the appropriately substituted benzimidazole (Examples 160, 162, 163) or benzotriazole (Example 161) following the protocol of general method A.

[0449] biological activity

[0450] In vitro biological activity

[0451] (a) BRAF, CRAF and ARAF kinase activity assays

[0452] Compound preparation: Solid samples of each material in 1 dram vials were suspended in DMSO (Fisher Scientific) at a stock concentration of 20 mM. Stocks were stored at -20°C and protected from light. If compound solubility at 20 mM was deemed problematic, the initial concentration of the DMSO stock was altered to 10 mM or 5 mM.

[0453] In vitro enzyme reactions were used to assess compound specific activity against BRAF, CRAF, and ARAF. For BRAF and CRAF, 0.375 nM purified GST-tagged kinase (Millipore Sigma, Cat. No. B4062-10UG and Cat. No. R1656-10UG, respectively) was incubated 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 a buffer containing 50 mM HEPES (pH 7.5), 10 mM MgCl2, 1 mM EDTA, 0.01% Brij-35, 2 mM DTT, with or without test compound. Separate reactions were performed with MEK1 substrate and ATP as a blank control. The ARAF kinase reaction was exactly the same except that the kinase concentration was increased to 3.75 nM (catalog no. 1768-0000-1; Reaction Biology).

[0454] For compound treatment, 5 μL / well of the test substance solution is placed in a 384-well proxyplate (Perkin Elmer) and mixed with 2-fold concentrated kinase reaction solution. A dilution series is selected such that 9 concentrations cover the range from 100 nM to 0.01 nM. If necessary (when the compound shows low intrinsic potency), the initial concentration of 100 nM is changed to 1 μM or 0.5 μM, and further dilutions are made accordingly. The final concentration of DMSO in the assay is set to 0.05%.

[0455] BRAF and CRAF kinase reactions were carried out at 30°C for a total of 2 hours and then stopped by diluting 1 / 2 with ADP-Glo ​​Reagent (Cat. No. V9102; Promega; part V912C). Reactions were then incubated at room temperature for 1 hour before adding 1 volume of Kinase Detection Reagent (Cat. No. V9102; Promega; part V917A). Plates were then equilibrated at room temperature for 30 minutes before detecting luminescence with a Synergy Neo2 plate reader (Biotek). The effect of each compound dilution on BRAF and CRAF kinase activity was expressed as % inhibition and calculated as follows: First, an internal 100% inhibition control (average of luminescence in kinase reactions containing only kinase-dead MEK1 substrate) was subtracted from each data point. The average of the DMSO (vehicle) controls (set as 0% inhibition) was set, and the following formula was used to calculate % inhibition:

number

[0456] The ARAF kinase reaction was carried out for a total of 2 hours at 30° C. and then stopped by adding EDTA to a final concentration of 40 mM. (登録商標) SureFire (登録商標) Ultra 商標 Detection was performed using the p-MEK 1 / 2 (Ser218 / 222) (Perkin Elmer) kit. Reactions were performed in 384-well proxy plates (Perkin Elmer) with 5 μL of kinase according to the manufacturer's specifications, followed by incubation of the reaction overnight at room temperature in a humidified chamber. After completion of the detection reaction, the AlphaLISA (登録商標)Signals were recorded on a Synergy Neo2 plate reader (Biotek) equipped with filters. The effect of each compound dilution on the pMEK signal generated by the ARAF reaction was expressed as % inhibition and calculated using the following formula: To measure pMEK background, an internal 100% inhibition control (average of luminescence in kinase reactions containing only kinase-dead MEK1 substrate) was included in each plate and subtracted from each data point. The average of the DMSO (vehicle) controls (set as 0% inhibition) was also set and used to calculate % inhibition.

number

[0457] I C 50 Values ​​were obtained by plotting kinase inhibition values ​​and fitting dose-activity curves with a log(agonist) vs. response-variable slope (four parameter) function using GraphPadPrism (V7.0) or the Dotmatics Screening Ultra platform. Standards included in the ARAF kinase assay were Belvarafenib (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 (Cat. No. HY-18957; CAS No. 1446090-79-4).

[0458] Thus, 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 2-5 below, while the ARAF kinase inhibitory potencies of representative analogs are listed in Table A below. Preferred examples, as defined in the embodiments, are BRAF IC 50 values ​​<10 nM, and even more preferred embodiments show BRAF IC 50 A preferred embodiment as defined in the embodiments has a CRAF IC 50 A more preferred embodiment shows a CRAF IC value <50 nM. 50 has a value <10 nM.

[0459] [Table A]

[0460] For ARAF biochemical kinase assay, * indicates IC 50 >20nM, ** indicates IC of 10-20nM 50 and *** indicates the range of IC 50 <10nM.

[0461] (b) Common cell culture methods

[0462] All cancer cell lines (A375, A101D, A2058, RKO, HT29 SK-MEL 30, IPC298, HepG2, HCT-116, Lovo, SW620, SW480, NCI-H358, NCI-H2122, Calu-6, NCIH2087, NCIH1755, NCIH1666, and Mewo) were obtained from ATCC and cultured in RPMI-1640 medium (Gibco) supplemented with 5% heat-inactivated fetal bovine serum (FBS, Wisent) at 37°C and 5% CO2. Cells were maintained in T175 flasks (Greiner). The culture medium was removed, washed once with 10 mL of room temperature phosphate-buffered saline (PBS; Wisent), and passaged by incubation with 2 mL of 0.05% trypsin (Thermo-Fisher) at 37°C. Trypsin was then inactivated by adding complete growth medium, and cells were replated in T175 culture dishes at the appropriate dilution. All cell lines were routinely tested for mycoplasma contamination. The tissue type and mutation status of each cell line can be seen in Table B below.

[0463] [Table B]

[0464] (c)AlphaLISA (登録商標) SureFire (登録商標) Ultra 商標 Measurement of phospho-ERK inhibition in cultured human cancer cell lines by p-ERK 1 / 2 (Thr202 / Tyr204)

[0465] AlphaLISA (登録商標) SureFire (登録商標) Ultra 商標 p-ERK 1 / 2 (Thr202 / Tyr204) analysis was performed on cells seeded in 100 μL of complete RPMI-1640 growth medium in 96-well flat-bottom clear dishes (Costar) at the densities indicated in Table C below. Cells were maintained overnight at 37° C. under 5% CO2 before being treated with a dilution series of compounds for 1 hour. Cell density (cells / cm2 ) is the number of cells per well of a 96-well plate (0.143 cm 2 ) is equivalent to dividing it by

[0466] [Table C]

[0467] In the dilution series, 100 μL / well of test substance dilutions prepared in complete RPMI-1640 growth medium are added to cells. The dilution series is selected to cover the range of 30 μM or 10 μM to 0.33 nM with 10 concentrations. If necessary (as in the case of A375 cells and NCIH1666 cells, which are generally more sensitive to compounds), the initial concentration of 10 μM is increased to 100 μM or decreased to 1 μM, and further dilutions are made accordingly. The final concentration of DMSO in the assay is set to 0.5%.

[0468] After treatment, the medium was removed and the cells were lysed in 50 μL of 1×AlphaScreen Ultra Lysis Buffer (Perkin Elmer). (登録商標) SureFire (登録商標) Ultra 商標 p-ERK 1 / 2 (Thr202 / Tyr204) (Perkin Elmer) reactions were performed in 384-well proxyplates (Perkin Elmer) according to the manufacturer's specifications with 5 μL of cell lysate, followed by incubation of the reactions overnight at room temperature in a humidified chamber. After completion of the reactions, the results were analyzed using the built-in AlphaLISA on an EnVision plate reader (Perkin Elmer). (登録商標) The signal was recorded using the settings.

[0469] The effect of each compound dilution on the pERK signal was expressed as % inhibition and calculated with the following formula: An internal 100% inhibition control (1 mM trametinib, Catalog No. HY-10999; MedChem Express; CAS No. 871700-17-3) was included in each plate and used as a measure of pERK background. First, the value obtained for trametinib was subtracted from each data point. The mean value of the DMSO (vehicle) control (set as 0% inhibition) was set and used to calculate the % inhibition.

number

[0470] The ability of each compound to inhibit pERK signaling was determined by plotting the inhibition values ​​for each data point in the dilution series and fitting the resulting curves with a log(agonist) vs. response-variable slope (four parameter) function using GraphPad Prism (V7.0). 50 expressed as a value.

[0471] If present, paradoxical pERK induction is determined by the pERK IC of the compound 50 The negative inhibition percentage values ​​observed in the curves were inferred. To classify a compound as a pERK paradoxical inducer, the polymer content of the inhibition percentage of the minimum data point of the dose-activity curve (Y 最小 %) was set to less than -20%, which is considered within the expected assay variation (e.g., Y 最小 Compounds with %=-30%, -50%, or -150% are considered to produce paradoxical induction of the pathway. 最小 =-10% IC 50 (Compounds that exhibit a curve are considered not to cause paradoxical activation of the pathway.) Therefore, compounds that meet the following criteria in a given cell line are said to inhibit the pathway without causing paradoxical induction: 1. The inhibition percentage at the highest dose (30 μM, 10 μM or 1 μM) was over 50%. 2.IC 50 Y of the station line 最小 % was greater than or equal to -20%. 最小 is the IC of the compound 50 It corresponds to the data point with the lowest value on the curve.

[0472] It is well known to those skilled in the art that some variability in inhibition values ​​is to be expected in such experiments. 最小 Values ​​of ±20% are considered within experimental error and are not significant. Therefore, only compounds with negative values ​​(greater than about 20%) beyond the variability of the assay are considered to induce paradoxical activation of the signaling cascade and are not within the scope of this disclosure. Figure 1 shows the IC of a compound that induces paradoxical pathway activation (PLX4720, commercially available from Selleck Chemical; CAS number 918505-84-7) and representative compounds described herein that exhibit an unexpected and distinct induction-free profile. 50 Provide a visualization of the curve.

[0473] FIG. 1 shows that RAS mutant HCT116 cells (Examples 80 and 81) do not induce paradoxical induction of pERK signaling (Y 最小 >-20%) compared with the compounds described herein, which showed strong induction of the pathway (Y 最小 Representative IC for compound (PLX4720) that causes ~-600%) 50 Inhibitory dose-response curves are shown.

[0474] Importantly, the compounds defined herein do not induce paradoxical activation of the pathway according to the criteria set forth above. To further illustrate this highly desirable property, the results of a non-inducing compound (Example 80) and an inducer, PLX4720, can be shown using immunoblot analysis as described below and depicted in FIG. 2.

[0475] For immunoblot analysis, 500,000 HCT-116 cells were seeded in 1 mL of complete RPMI-1640 growth medium in 24-well flat-bottom clear dishes (Costar). Cells were maintained overnight at 37°C under 5% CO2 before being treated with a dilution series of compounds for 1 h. Cells were then washed once with PBS and lysed for 15 min at 4°C with gentle rocking in 250 μL of Igepal Lysis Buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1% Igepal-CA630, 1 mM EDTA, 10% glycerol) supplemented with leupeptin, aprotinin, PMSF, phosphatase inhibitor cocktail (Sigma), and Na3VO4. Cell extracts were then clarified by centrifugation at 20,000g for 10 min at 4°C. The cleared lysates were transferred into fresh tubes on ice and then boiled for 5 min in sample loading buffer (100 mM Tris-HCl, pH 6.8, 4% SDS, 0.2% bromophenol blue, 20% glycerol, 200 mM β-mercaptoethanol) before fractionation by SDS-PAGE and transfer to nitrocellulose membranes (PALL). Membranes were blocked for 1 h in Tris Buffered Saline 0.2% Tween-20 (TBST; 10 mM Tris-HCl, pH 8.0, 0.2% Tween-20, 150 mM NaCl) containing 2% BSA (Sigma) and then incubated overnight at 4° C. with the following dilutions of primary antibodies prepared in TBST: anti-pERK (1:2000 dilution; Sigma-Aldrich; Cat. No. M9692), all anti-ERK (1:1000 dilution; Cell Signaling Technology; Cat. No. 4695), anti-pMEK (1:1000 dilution; Cell Signaling Technology; Cat. No. 9121), and all anti-MEK (1:1000 dilution; Cell Signaling Technology; Cat. No. 9122).Secondary anti-mouse-HRP and anti-rabbit-HRP (Jackson Immunoresearch Labs; Cat. Nos. 115-035-146 and 111-035-144, respectively) were prepared at 1:5000 and 1:10000 dilutions, respectively, in TBST. Immunoblots were visualized by exposure to X-ray film after 1 min incubation with ECL reagent.

[0476] Figure 2 shows the results of immunoblot analysis of RAS mutant HCT-116 cells treated with a representative compound that does not induce paradoxical induction of pERK or pMEK signals (Example 80; top panel) and, in comparison, a compound that induces the pathways in the same cell line (PLX4720; bottom panel). Total MEK and total ERK signals were also probed by immunoblot to ensure equal loading of protein samples between conditions. Compound concentrations are shown above the immunoblot panels in micromolar. The concentration range used for treatment was the same for Example 80 and PLX4720.

[0477] The compounds of the examples 1-163 exhibit pERK inhibitory activity in the colon G13D Ras mutated HCT-116 cell line, as shown in Tables 2-5. In addition, some examples have also been shown to exhibit inhibition without paradoxical induction of pERK signaling in the SW480 colon cancer cell line carrying the G12D allele of KRAS (Tables 2-5). Moreover, some examples in Tables 2-5 show inhibition of BRAF signaling. V600E Driver mutation (BRAF V600E The compounds were also tested for inhibition of pERK in A375 cells containing a pERK driver mutation and were found to be similarly active (Table D). In Tables 2-5, all compounds, Examples 1-163, had pERK IC values ​​of less than 30 μM in the HCT116 cell line. 50 The preferred compounds are 50 The IC value is 1-10 μM, and more preferred compounds are 50The IC value is 0.5 to 1 μM, and more preferred compounds are 50 The value was less than 0.5 μM.

[0478] Representative compounds defined herein were also tested for their pERK inhibitory activity against additional tumor cells and showed good to very good pERK inhibitory activity in cancer cell lines harboring various NRAS, KRAS and NF1 alleles and representing a wide variety of tissue types (i.e., SK-MEL 30, IPC298, HepG2, HCT-116, Lovo, SW620, SW480, NCI-H358, NCI-H2122, Calu-6 and Mewo; see Table D below and Table B for genotypes). The pERK inhibitory activity of the compounds was stronger in cancer cell lines harboring BRAF alleles (A375, A101D, A2058, RKO, HT29, NCIH2087, NCIH1755 and NCIH1666) (Table D).

[0479] Table D (Tables D-1 and D-2). Panel of RAS mutant cancer cell lines (see Table B for genotypes) and BRAF V600E No inducible pERK IC for selected compounds in mutant A375 50 Value and antiproliferative EC 50 value.

[0480] [Table D-1]

[0481] In pERK, + is IC 50 >300nM, ++ indicates IC of 30-300nM 50 and +++ indicates the range of IC 50 <30 nM. For proliferation, * indicates EC 50 EC > 3000nM, ** indicates EC of 300-3000nM 50 *** indicates the range of EC 50 <300 nM is indicated. Belv: belvalafenib. Values ​​in brackets are % inhibition. Blanks mean that values ​​were not determined.

[0482] [Table D-2]

[0483] In pERK, + is IC 50 >300nM, ++ indicates IC of 30-300nM 50 and +++ indicates the range of IC 50 <30 nM. For proliferation, * indicates EC 50 EC > 3000nM, ** indicates EC of 300-3000nM 50 *** indicates the range of EC 50 <300 nM is indicated. Belv: belvalafenib. Values ​​in brackets are % inhibition. Blanks mean that values ​​were not determined.

[0484] In the case of RAS mutant cancer cell lines, pERK IC 50 Curve Y 最小 All % values ​​were above -20% and were considered to show minimal or no induction, and therefore the compound does not cause detectable paradoxical activation of the pathway in this panel of cancer cell lines. In contrast, comparative results for belbalafenib (obtained from MedChem Express, catalog number HY-109080; CAS number 1446113-23-0) show mild to strong pathway induction in the same cell lines (Y in 10 of the 13 RAS mutant cell lines tested). 最小 <-30%).

[0485] (d) CellTiter-Glo (登録商標) Measurement of the growth inhibition effect of cultured human cancer cell lines (CCL) using the reagent

[0486] CellTiter-Glo (登録商標) Viability analysis was performed on cells seeded in 96-well flat-bottom white opaque plates (Greiner or Corning) in 40 μL of complete RPMI-1640 growth medium at the densities indicated in Table E below (for each CCL, CellTiter-Glo (登録商標)(number of cells seeded per well of a 96-well plate to perform cell viability assays). Cell density (cells / cm 2 ) is the number of cells per well of a 96-well plate (0.32 cm 2 ) Cells were maintained overnight at 37° C. under 5% CO2 before being treated with a dilution series of compounds for 3 days.

[0487] [Table E]

[0488] In the dilution series, 100 μL / well of test substance dilutions prepared in complete RPMI-1640 growth medium are added to cells seeded in 100 μL growth medium first. The dilution series is selected to cover the range of 30 μM or 10 μM to 0.33 μM with 10 concentrations. If necessary (such as in the case of A375 cells, which are more sensitive to compounds), the initial concentration of 10 μM is reduced to 1 μM and further dilutions are made accordingly. The final concentration of DMSO in the assay is set to 0.5%.

[0489] After 3 days of incubation, the growth medium was removed by aspiration, and 60 μL of diluted CellTiter-Glo was added. (登録商標) Reagent (10 μL of CellTiter-Glo (登録商標) Reagent + 50 μL of PBS) was added to each well. Cells were lysed and stained with CellTiter-Glo by incubating on a plate shaker for 5 minutes. (登録商標) After equilibration in reagent and subsequent incubation at room temperature for 10 min, luminescence signals were then acquired on a Synergy Neo2 plate reader (Biotek).

[0490] The effect of each compound dilution on the proliferation of the cancer cell lines was expressed as % inhibition and calculated using the formula shown below: An internal 100% inhibition control (1 mM trametinib; Catalog No. HY-10999; MedChem Express; CAS No. 871700-17-3) was included in each plate and analyzed by CellTiter-Glo. (登録商標) This was used as a measure of the signal background. The value obtained for trametinib was subtracted from each data point. The mean value of the DMSO (vehicle) control (set as 0% inhibition) was set and used to calculate the % inhibition.

number

[0491] The growth inhibitory potency of each compound was determined as an EC200 expression level by plotting the effect value for each data point in the dilution series using GraphPad Prism (V7.0) or the Dotmatics Screening Ultra platform, and fitting the resulting curves with a log(agonist) vs. response-variable slope (four parameter) function. 50 expressed as a value.

[0492] As shown in Table D, the active agents exhibit antiproliferative activity in a variety of different NRAS-, KRAS- and NF1-mutated cancer cell lines (i.e., SK-MEL 30, IPC298, HepG2, HCT-116, Lovo, SW620, SW480, NCI-H358, NCI-H2122, Calu-6 and Mewo; see Table D and for genotypes see Table B) that represent a large diversity of tissue types. Antiproliferative activity is often even stronger in cell lines with B BRAF driver mutations (Table D). Notably, the IC of pERK reduction in KRAS and BRAF mutant cell lines was 0.018-0.028. 50 Values ​​and EC of the antiproliferative activity of substances 50The values ​​correlate well with each other (Table D). Thus, the compounds of the present invention are effective against several tumor types and may be used for these and other indications. This demonstrates that the compounds defined herein are useful for the treatment of various types of tumors.

[0493] (e) result

[0494] Tables 2-5 below summarize the structures, synthetic methods, and biological results of exemplary compounds. Each of these tables is followed by its respective table summarizing the chemical characterization of the compounds.

[0495] [Table 2] JPEG2024517505000123.jpg244170

[0496] For pERK assays, + indicates IC of 10–30 μM 50 ++ indicates IC of 1-10 μM 50 +++ indicates IC of 0.5-1 μM 50 and ++++ indicates the range of IC 50 <0.5 μM. 最小 The percentage values ​​are for each IC 50 Indicates the lowest point of the curve. 最小 IC with value > -20% 50 Compounds showing a curve show minimal or no induction and are considered not to cause detectable paradoxical activation of the pathway. For the BRAF biochemical kinase assay, * denotes IC 50 >10nM, ** indicates IC of 1-10nM 50 indicates the range of IC 50 <1 nM. For the CRAF biochemical kinase assay, § indicates IC 50 >50 nM, and §§ indicates IC of 10-50 nM. 50 §§§ indicates the range of IC 50 <10nM.

[0497] [Table 2-1] JPEG2024517505000125.jpg234170JPEG2024517505000126.jpg214170

[0498] [Table 3] JPEG2024517505000128.jpg235170JPEG2024517505000129.jpg237170JPEG2024517505000130.jpg136170

[0499] For pERK assays, + indicates IC of 10–30 μM 50 ++ indicates IC of 1-10 μM 50 +++ indicates IC of 0.5-1 μM 50 and ++++ indicates the range of IC 50 <0.5 μM. 最小 The percentage values ​​are for each IC 50 Indicates the lowest point of the curve. 最小 IC with value > -20% 50 Compounds showing a curve show minimal or no induction and are considered not to cause detectable paradoxical activation of the pathway. For the BRAF biochemical kinase assay, * denotes IC 50 >10nM, ** indicates IC of 1-10nM 50 indicates the range of IC 50 <1 nM. For the CRAF biochemical kinase assay, § indicates IC 50 >50 nM, §§ indicates an IC50 range of 10-50 nM, and §§§ indicates an IC50<10 nM.

[0500] [Table 3-1] JPEG2024517505000132.jpg218170JPEG2024517505000133.jpg226170JPEG2024517505000134.jpg215170JPEG20245175050 00135.jpg220170JPEG2024517505000136.jpg229170JPEG2024517505000137.jpg226170JPEG2024517505000138.jpg220170 JPEG2024517505000139.jpg231170JPEG2024517505000140.jpg217170JPEG2024517505000141.jpg229170JPEG20245175050 00142.jpg215170JPEG2024517505000143.jpg234170JPEG2024517505000144.jpg231170JPEG2024517505000145.jpg219170

[0501] [Table 4]

[0502] For pERK assays, + indicates IC of 10–30 μM 50 ++ indicates IC of 1-10 μM 50 +++ indicates IC of 0.5-1 μM 50 and ++++ indicates the range of IC 50 <0.5 μM. 最小 The percentage values ​​are for each IC 50 Indicates the lowest point of the curve. 最小 IC with value > -20% 50 Compounds showing a curve show minimal or no induction and are considered not to cause detectable paradoxical activation of the pathway. For the BRAF biochemical kinase assay, * denotes IC 50 >10nM, ** indicates IC of 1-10nM 50 indicates the range of IC 50 <1 nM. For the CRAF biochemical kinase assay, § indicates IC 50>50 nM, and §§ indicates IC of 10-50 nM. 50 §§§ indicates the range of IC 50 <10nM.

[0503] [Table 4-1]

[0504] [Table 5]

[0505] For pERK assays, + indicates IC of 10–30 μM 50 ++ indicates IC of 1-10 μM 50 +++ indicates IC of 0.5-1 μM 50 and ++++ indicates the range of IC 50 <0.5 μM. 最小 The percentage values ​​are for each IC 50 Indicates the lowest point of the curve. 最小 IC with value > -20% 50 Compounds showing a curve show minimal or no induction and are considered not to cause detectable paradoxical activation of the pathway. For the BRAF biochemical kinase assay, * denotes IC 50 >10nM, ** indicates IC of 1-10nM 50 indicates the range of IC 50 <1 nM. For the CRAF biochemical kinase assay, § indicates IC 50 >50 nM, and §§ indicates IC of 10-50 nM. 50 §§§ indicates the range of IC 50 <10nM.

[0506] [Table 5-1]

[0507] Numerous modifications can be made to any of the above described embodiments without departing from the scope of the invention. Any literature, patent or scientific literature referred to herein is incorporated by reference in its entirety for all purposes.

Claims

1. A compound of formula I: 【Chemistry 1】 Where: R 1 is replaced or not replaced, OR 3 , S.R. 3 , N.H. 2 , N.H.R. 3 , N(R 3 ) 2 , C 3～8 Cycloalkyl, C 4～8 Heterocycloalkyl, C 6～10 Aryl and C 5～10 heteroaryl; R 2 is replaced by C 6 Aryl and C 5～10 Heteroaryl, substituted or unsubstituted, C 4～8 Heterocycloalkyl and N(R 3 ) 2 Selected from: R 3 is independently at each occurrence substituted or unsubstituted; C 1～8 Alkyl, C 3～8 Cycloalkyl, C 4～8 Heterocycloalkyl, C 6～10 Aryl and C 5～10 heteroaryl; X 1 is a halogen atom or an electron-withdrawing group; X 2 is selected from H, a halogen atom, and an electron withdrawing group; X 3 and X 4 are H, halogen atoms, electron-withdrawing groups, and C 1～3 Alkyl, C 3～4 Cycloalkyl and OC 1～3 alkyl; Y is H, halogen atom, CN, OH, OC 1～8 Alkyl, NH 2 , N.H.C. 1～8 Alkyl, N(C 1～8 Alkyl) 2 , and substituted or unsubstituted C 1～8 alkyl; However, the compounds in question are as follows: 【Chemistry 2】

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

3. R 2 The compound according to claim 1, wherein is a group of the formula 【Chemistry 4】 Where: X 5 NH, NC 1～3 Alkyl, NC 3～4 cycloalkyl, O and S; R 9 , R 10 , R 11 are each independently H, F, Cl, CN, and substituted or unsubstituted, C 1～3 Alkyl, C 3～4 Cycloalkyl, C(O)OC 1～3 Alkyl or OC 1～3 alkyl, where R 9 and R 11 is H and the other is different from H; and (---) represents a bond.

4. R 2 The compound according to claim 1, wherein is a group of the formula 【Chemistry 5】 Where: X 5 NH, NC 1～3 Alkyl, NC 3～4 cycloalkyl, O and S; R 9 is F, Cl, CN, and substituted or unsubstituted, C 1～3 Alkyl, C 3～4 Cycloalkyl, C(O)OC 1～3 Alkyl or OC 1～3 alkyl; R 10 and R 12 are each independently H, F, Cl, CN, and substituted or unsubstituted, C 1～3 Alkyl, C 3～4 Cycloalkyl, C(O)OC 1～3 Alkyl or OC 1～3 alkyl; and (---) represents a bond.

5. R 2 The compound according to claim 1, wherein is a group of the formula 【Chemistry 6】 Where: X 9 , X 10 , X 11 , X 12 and X 13 is independently selected from N and C, where X 9 , X 10 , X 11 , X 12 and X 13 at least one and not more than two of are N; and R 19 , R 20 , R 21 , R 22 and R 23 H, F, Cl, Br, CN, NO 2 , N.H. 2 , and substituted or unsubstituted, C 1～3 Alkyl, C 3～4 Cycloalkyl or OC 1～3 alkyl or their combined X 9 , X 10 , X 11 , X 12 Or X 13 does not exist when is N; where X 9 and X 13 is not N; and where X 9 and X 13 When at least one of the groups is N, the other group is not N or CH. 。

6. R 2 The compound according to claim 1, wherein is a group of the formula 【Chemistry 7】 Where: R 13 independently at each occurrence, F, Cl, and substituted or unsubstituted, C 1～3 Alkyl, C 3～4 Cycloalkyl or C 1～3 alkoxy; n is an integer selected from 0 to 8; or n is 2 to 8, and two R 18 together with their adjacent carbon atoms, C 3～4 forms a cycloalkyl; and (---) represents a bond.

7. R 2 N(R 3 ) 2 2. The compound of claim 1, wherein R 3 is selected from substituted or unsubstituted C 1-8 alkyl or C 3-8 cycloalkyl.

8. R 2 The compound according to claim 1, wherein is selected from the groups B1 to B77 below. (---) represents a bond.

9. R 1 2. The compound of claim 1, wherein is a substituted or unsubstituted group selected from imidazolyl, pyrazolyl, triazolyl, indolyl, indazolyl, benzimidazolyl, benzotriazolyl, pyrrolopyridinyl, pyrazolopyridinyl, purinyl, and imidazopyrazinyl.

10. 10. The compound of claim 9, wherein the substituted or unsubstituted groups are attached to the pyrimidopyrimidine core through a nitrogen atom.

11. R 1 OH, halogen atoms, CN, NO 2 , C 1～6 Alkyl, C 2～6 Alkenyl, C 2～6 Alkynyl, OC 1～6 Alkyl, C 5～10 Heteroaryl, C 3～10 Cycloalkyl, C 4～10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 ) 2 , S.O. 2 R 15 , S.O. 2 N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , N(R 16 )C(O)N(R 14 ) 2 , N(R 16 )SO 2 N(R 14 ) 2 , N(R 14 ) 2 , P(O)(R 15 ) 2 , C.H. 2 C(O)R 15 , C.H. 2 C(O)N(R 14 ) 2 , C.H. 2 SO 2 R 15 , C.H. 2 SO 2 N(R 14 ) 2 , C.H. 2 N(R 16 )C(O)R 15 , C.H. 2 N(R 16 )SO 2 R 15 , C.H. 2 N(R 16 )C(O)N(R 14 ) 2 , C.H. 2 N(R 16 )SO 2 N(R 14 ) 2 , and C.H. 2 N(R 14 ) 2 and is substituted with at least one substituent selected from Where: R 14 independently in each case, 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, C 4～10 forming a heterocycloalkyl group, wherein said alkyl, said alkenyl, said alkynyl, said cycloalkyl, said heterocycloalkyl, said aryl or said heteroaryl group may be further substituted; R 15 In each case, 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 said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group may be further substituted; and R 16 independently in each case, H, C 1～6 Alkyl, C 2～6 Alkenyl, C 2～6 Alkynyl, C 3～10 Cycloalkyl, C 6 Aryl and C 5～10 and heteroaryl, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group may be further substituted. The compound of claim 1.

12. R 1 The compound according to claim 1, wherein is a group of the formula 【Chemistry 10】 Where: R 17 H, OH, halogen atoms, CN, NO 2 , C 1～6 Alkyl, C 2～6 Alkenyl, C 2～6 Alkynyl, OC 1～6 Alkyl, C 5～10 Heteroaryl, C 3～10 Cycloalkyl, C 4～10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 ) 2 , S.O. 2 R 15 , S.O. 2 N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , N(R 16 )C(O)N(R 14 ) 2 , N(R 16 )SO 2 N(R 14 ) 2 , N(R 14 ) 2 , P(O)(R 15 ) 2 , C.H. 2 C(O)R 15 , C.H. 2 C(O)N(R 14 ) 2 , C.H. 2 SO 2 R 15 , C.H. 2 SO 2 N(R 14 ) 2 , C.H. 2 N(R 16 )C(O)R 15 , C.H. 2 N(R 16 )SO 2 R 15 , C.H. 2 N(R 16 )C(O)N(R 14 ) 2 , C.H. 2 N(R 16 )SO 2 N(R 14 ) 2 , and C.H. 2 N(R 14 ) 2 Selected from: X 6 is N or CH; and X 7 is N, and R 18 is not present; or X 7 is C, and R 18 But, C 1～6 Alkyl, C 2～6 Alkenyl, C 2～6 Alkynyl, OC 1～6 Alkyl, C 5～10 Heteroaryl, C 3～10 Cycloalkyl, C 4～10 Heterocycloalkyl, C(O)R 15 , C(O)N(R 14 ) 2 , S.O. 2 R 15 , S.O. 2 N(R 14 ) 2 , N(R 16 )C(O)R 15 , N(R 16 )SO 2 R 15 , N(R 16 )C(O)N(R 14 ) 2 , N(R 16 )SO 2 N(R 14 ) 2 , N(R 14 ) 2 , P(O)(R 15 ) 2 , C.H. 2 C(O)R 15 , C.H. 2 C(O)N(R 14 ) 2 , C.H. 2 SO 2 R 15 , C.H. 2 SO 2 N(R 14 ) 2 , C.H. 2 N(R 16 )C(O)R 15 , C.H. 2 N(R 16 )SO 2 R 15 , C.H. 2 N(R 16 )C(O)N(R 14 ) 2 , C.H. 2 N(R 16 )SO 2 N(R 14 ) 2 , and C.H. 2 N(R 14 ) 2 Selected from: Where: R 14 is independently at each occurrence selected 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 atom form a C 4-10 heterocycloalkyl group, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group may be further substituted; R 15 is independently at each occurrence 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, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group may be further substituted; and R 16 is independently at each occurrence selected 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 said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, or heteroaryl may be further substituted; ;and, Here, (---) represents a bond.

13. R 1 The compound according to claim 1, wherein is a group of the formula 【Chemistry 11】 Where: R 14 is independently at each occurrence selected 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 atom form a C 4-10 heterocycloalkyl group, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group may be further substituted; and R 15 is independently at each occurrence 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, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group may be further substituted; Here, (---) represents a bond.

14. R 1 The compound according to claim 1, wherein is a group of the formula 【Chemistry 13】 Where: X 15 , X 16 , X 17 , and X 18 are independently O, N, S and CR 17 is selected from where R 17 is H, OH, a halogen atom, 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(R14)2, CH2N(R16)SO2N(R14)2, and CH2N(R14)2; Where: R 14 is independently at each occurrence selected 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 atom form a C 4-10 heterocycloalkyl group, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group may be further substituted; R 15 is independently at each occurrence 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, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group may be further substituted; and R 16 is independently at each occurrence selected 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 said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group may be further substituted; where X 15 , X 16 , X 17 and X 18 Up to two of these are O, N, or S.

15. R 1 The compound according to claim 1, wherein is selected from the groups C1 to C493 listed below.

16. The compound according to claim 1, wherein the compound is a compound of formula II: 【Chemistry 14】 Where: R 1 is as defined above, 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; 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 is 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; or R 4 and R 5 or R 5 and R 6 together with their adjacent carbon atoms form a substituted or unsubstituted carbocyclic or heterocyclic ring, provided that the heterocyclic ring is not a benzoxazolinone; wherein when R 4 is H or F, then at least one of R 5 or R 6 is other than H or F; and wherein when R 5 is CN, then at least one of R 4 or R 6 is other than H.

17. The compound according to claim 1, wherein the compound is a compound of formula IV: 【Chemistry 15】 Where: X 6 is N or CH; and X 7 is N and R 18 is absent; or X 7 is C and R 18 is 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, CH2N(R16)SO2N(R14)2, and CH2N(R14)2; 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; 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 is 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; or R 4 and R 5 or R 5 and R 6 together with their adjacent carbon atoms form a substituted or unsubstituted carbocyclic or heterocyclic ring, provided that the heterocyclic ring is not a benzoxazolinone; wherein when R 4 is H or F, then at least one of R 5 or R 6 is other than H or F; and wherein when R 5 is CN, then at least one of R 4 or R 6 is other than H; R 17 is H, OH, a halogen atom, 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, CH2N(R16)SO2N(R14)2, and CH2N(R14)2; R 18 is 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 2C(O)N(R14)2, CH2SO2R15, CH2SO2N(R14)2, CH2N(R16)C(O)R15, CH2N(R16)SO2R15, CH2N(R16)C(O)N(R14)2, CH2N(R16)SO2N(R14)2, and CH2N(R14)2; R 14 is independently at each occurrence selected 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 atom form a C 4-10 heterocycloalkyl group, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group may be further substituted; R 15 is independently at each occurrence 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, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group may be further substituted; and R 16 is independently at each occurrence selected 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 said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group may be further substituted.

18. The compound according to claim 1, wherein the compound is a compound of formula V: 【Chemistry 16】 Where: 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; 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 is 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; or R 4 and R 5 or R 5 and R 6 together with their adjacent carbon atoms form a substituted or unsubstituted carbocyclic or heterocyclic ring, provided that the heterocyclic ring is not a benzoxazolinone; wherein when R 4 is H or F, then at least one of R 5 or R 6 is other than H or F; and wherein when R 5 is CN, then at least one of R 4 or R 6 is other than H; X 15 , X 16 , X 17 , and X 18 are independently selected from O, N, S, and CR 17 , where no more than two of X 15 , X 16 , X 17 , and X 18 are O, N, or S; where R 17 is H, OH, a halogen atom, 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(R14)2, CH2N(R16)SO2N(R14)2, and CH2N(R14)2; R 14 is independently at each occurrence selected 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 atom form a C 4-10 heterocycloalkyl group, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group may be further substituted; R 15 is independently at each occurrence 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, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group may be further substituted; and R 16 is independently at each occurrence selected 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 said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group may be further substituted.

19. The compound according to claim 1, wherein the compound is a compound of formula III: 【Chemistry 17】 Where: R 1 is as defined above, R 9 , 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, with the proviso that one of R 9 and R 11 is H and the other is different from H; X 5 is selected from NH, NC 1-3 alkyl, NC 3-4 cycloalkyl, O and S.

20. The compound according to claim 1, wherein the compound is a compound of formula VI: 【Chemistry 18】 Where: R 9 , 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, with the proviso that one of R 9 and R 11 is H and the other is different from H; R 17 is H, OH, a halogen atom, 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, CH2N(R16)SO2N(R14)2, and CH2N(R14)2; X5 is selected from NH, NC1-3alkyl, NC3-4cycloalkyl, O and S; X 6 is N or CH; X 7 is N and R 18 is absent; or X 7 is C and R 18 is 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, CH2N(R16)SO2N(R14)2, and CH2N(R14)2; R 14 is independently at each occurrence selected 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 atom form a C 4-10 heterocycloalkyl group, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group may be further substituted; R 15 is independently at each occurrence 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, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group may be further substituted; and R 16 is independently at each occurrence selected 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 said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group may be further substituted.

21. The compound according to claim 1, wherein the compound is a compound of formula VII: 【Chemistry 19】 Where: R 9 , 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, with the proviso that one of R 9 and R 11 is H and the other is different from H; X5 is selected from NH, NC1-3alkyl, NC3-4cycloalkyl, O and S; X 15 , X 16 , X 17 , and X 18 are independently selected from O, N, S, and CR 17 , where no more than two of X 15 , X 16 , X 17 , and X 18 are O, N, or S; where R 17 is H, OH, a halogen atom, 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(R14)2, CH2N(R16)SO2N(R14)2, and CH2N(R14)2; R 14 is independently at each occurrence selected 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 atom form a C 4-10 heterocycloalkyl group, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group may be further substituted; R 15 is independently at each occurrence 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, wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group may be further substituted; and R 16 is independently at each occurrence selected 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 said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl group may be further substituted.

22. A pharmaceutical composition comprising a compound according to any one of claims 1 to 21, together with a pharma- ceutically acceptable carrier, diluent or excipient.

23. A compound according to any one of claims 1 to 21 for the treatment of a disease or disorder selected from a proliferative disease or disorder, developmental disorders caused by dysregulation of the RAS-ERK signalling cascade (RASopathies), or an inflammatory disease or immune system disorder.

24. 24. The compound of claim 23, wherein the disease or disorder is selected from neoplasms and developmental disorders.

25. The compound of claim 23, wherein the disease or disorder is associated with a RAF gene mutation or a RAS gene mutation.

26. 24. The compound of claim 23, wherein the disease or disorder is associated with a mutation or amplification of a receptor tyrosine kinase, or a mutation or amplification in a RAS downstream regulator of the receptor.

27. 25. The compound of claim 24, wherein the neoplasm is selected from melanoma, thyroid cancer, colorectal cancer, ovarian cancer, breast cancer, endometrial cancer, liver cancer, sarcoma, gastric cancer, pancreatic cancer, Barrett's adenocarcinoma, glioma, lung cancer, head and neck cancer, acute lymphocytic leukemia, acute myeloid leukemia, non-Hodgkin's lymphoma, and hairy cell leukemia.

28. An agent for use in treating a disease or disorder comprising a compound according to any one of claims 1 to 21, wherein the disease or disorder is selected from a proliferative disease or disorder, a developmental disorder caused by dysregulation of the RAS-ERK signaling cascade (RASopathies), or an inflammatory disease or immune system disorder.

29. The method of claim 28, wherein the disease or disorder is selected from neoplasms and developmental abnormalities.

30. The agent described in claim 28, wherein the disease or disorder is associated with a RAF gene mutation or a RAS gene mutation.

31. The agent described in claim 28, wherein the disease or disorder is associated with a mutation or amplification of a receptor tyrosine kinase, or a mutation or amplification in a RAS downstream regulator of the receptor.

32. The agent described in claim 28, wherein the neoplasm is selected from melanoma, thyroid cancer, colorectal cancer, ovarian cancer, breast cancer, endometrial cancer, liver cancer, sarcoma, gastric cancer, pancreatic cancer, Barrett's adenocarcinoma, glioma, lung cancer, head and neck cancer, acute lymphocytic leukemia, acute myeloid leukemia, non-Hodgkin's lymphoma, and hairy cell leukemia.

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