Cyclic compounds and methods of using same

Abstract

This application relates to compounds of formula (I) as defined herein, and pharma- ceutically acceptable salts thereof. This application also describes pharmaceutical compositions comprising the compounds of formula (I), and pharma- ceutically acceptable salts thereof, and methods of using the compounds and compositions for inhibiting kinase activity and treating cancer. Also provided herein is a method of inhibiting cell proliferation in vitro or in vivo, comprising contacting the cell with an effective amount of a compound of formula (I) as defined herein, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition thereof.

Description

[Technical Field]

[0001] This application relates to tricyclic and other polycyclic compounds that are useful in treating proliferative disorders such as cancer. [Background technology]

[0002] Cancer is characterized by abnormal cell growth and proliferation. Genomic instability is a hallmark of cancer cells, and high mutation rates and genomic rearrangements lead to invasive and treatment-resistant tumors. See Hanahan and Weinberg, Cell, 144, pp. 646-674 (2011), and McGranahan and Swanton, Cell, 168, pp. 613-628 (2017). Dysregulation of DNA replication contributes to genomic instability and tumorigenesis. Eukaryotic cells divide through a directed, highly regulated, step-by-step process known as the cell cycle. DNA replication is an integral part of the highly regulated, step-by-step cell cycle, and this strict regulation ensures that DNA replication occurs only once during S phase and with high fidelity.

[0003] During late G1-S phase, CDC7 kinase (also known as DDK) is activated by binding to its regulatory protein DBF4 (ASK in eukaryotes), which then phosphorylates chromatin-loaded minichromosome maintenance (MCM) 2, 4, and 6 proteins at multiple phosphorylation sites to initiate DNA synthesis. See Jiang et al., EMBO J., 18, pp. 5703-5713 (1999); Cho et al., Proc. Natl. Acad. Sci. USA, 103, pp. 11521-11526 (2006); and Masai et al., J. Biol. Chem., 281, pp. 39249-39261 (2006). CDC7 kinase plays an important role in maintaining DNA replication forks and the DNA damage response pathway. See Yamada, et al., Cell Cycle 13, pp. 1859-1866 (2014). CDC7 is a serine / threonine kinase highly conserved from yeast to humans. Knockdown of CDC7 has been shown to cause cell death in cancer cells, but not in normal cells where the p53-dependent pathway arrests the cell cycle at the G1 phase. The apoptotic response induced in cancer cells by CDC7 depletion is not mediated by p53, but rather by p38 MAPK. See Montagnoli, et al., Cancer Res., 64, pp. 7110-7116 (2004), and Im and Lee, J. Biol. Chem., 283, pp. 25171-25177 (2008). Furthermore, upregulation of CDC7 correlates with poor prognosis in various cancer types. See, for example, Kulkarni, et al., Clin. Cancer Res., 15, pp. 2417-2425 (2009); Choschzick, et al., Hum. Pathol., 41, pp. 358-365 (2010); Datta, et al., EMBO Rep., 18, pp. 2030-2050 (2017); Cheng, et al., Cancer Lett., 337, 218-225 (2013). [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Hanahan and Weinberg, Cell, 144, pp. 646-674 (2011) [Non-patent document 2] McGranahan and Swanton,Cell 168,pp.613-628(2017) [Non-patent document 3] Jiang,et al.,EMBO J.,18,pp.5703-5713(1999) [Non-patent document 4] Cho,et al.,Proc.Natl.Acad.Sci.USA,103,pp.11521-11526(2006) [Non-Patent Document 5] Masai,et al.,J Biol Chem.,281,pp.39249-39261(2006) [Non-patent document 6] Yamada,et al.,Cell Cycle 13,pp.1859-1866(2014) [Non-Patent Document 7] Montagnoli,et al.,Cancer Res.,64,pp.7110-7116(2004) [Non-patent document 8] Im and Lee, J.Biol.Chem.,283,pp.25171-25177(2008) [Non-Patent Document 9] Kulkarni,et al.,Clin.Cancer Res.,15,pp.2417-2425(2009) [Non-Patent Document 10] Choschzick,et al.,Hum. Pathol.,41,pp.358-365(2010) [Non-Patent Document 11] Datta,et al.,EMBO Rep.,18,pp.2030-2050(2017) [Non-Patent Document 12] , Cheng,et al.,Cancer Lett.,337,218-225(2013) Summary of the Invention [Means for solving the problem]

[0005] It has now been found that certain condensed compounds are inhibitors of CDC7 kinase and are useful in the treatment of diseases such as proliferative disorders, including cancer.

[0006] Thus, as used herein, compounds of formula (I): [ka] or a pharmaceutically acceptable salt thereof, wherein R 1, R 2 , R 3 , R 4 , ring A and m are as defined herein.

[0007] Also provided herein is a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

[0008] Also provided herein is a method of inhibiting proliferation of a cell in vitro or in vivo, comprising contacting the cell with an effective amount of a compound of formula (I), as defined herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0009] Also provided herein is a method of inhibiting CDC7 kinase activity in vitro or in vivo, comprising contacting a cell with an effective amount of a compound of formula (I), as defined herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0010] Also provided herein is a method of treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I), as defined herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0011] Also provided herein is a method of treating a CDC7-associated disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I), as defined herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0012] Also provided herein is a method of treating cancer in a subject in need thereof, comprising administering to a subject identified as having a cancer associated with CDC7 an effective amount of a compound of formula (I), as defined herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0013] Also provided herein is a method of treating cancer and / or inhibiting metastasis associated with a particular cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition, as defined herein.

[0014] There is also provided herein a compound of formula (I), as defined herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the treatment of cancer.

[0015] Also provided herein is a compound of formula (I), as defined herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the treatment of a CDC7-associated disease or disorder.

[0016] Also provided herein is a compound of formula (I), as defined herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the treatment of cancer and / or the inhibition of metastasis associated with certain cancers.

[0017] Also provided herein is a compound of formula (I), or a pharmaceutically acceptable salt thereof, for use in inhibiting CDC7 kinase activity.

[0018] Also provided herein is a compound of formula (I), as defined herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in the treatment of a CDC7-associated disease or disorder.

[0019] Also provided herein is a compound of formula (I), as defined herein, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating cancer and / or inhibiting metastasis associated with certain cancers.

[0020] Also provided herein is a compound of formula (I), as defined herein, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the inhibition of CDC7 kinase activity.

[0021] Also provided herein is a compound of formula (I), as defined herein, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a CDC7-associated disease or disorder.

[0022] Also provided are methods of treating an individual having a CDC7-associated cancer, comprising administering a compound of formula (I), or a pharmaceutically acceptable salt thereof, before, during, or after administration of another anti-cancer agent (e.g., a first CDC7 kinase inhibitor or another kinase inhibitor).

[0023] Also provided herein are processes for preparing compounds of formula (I) or pharmaceutically acceptable salts thereof.

[0024] Also provided herein are compounds of formula (I) or pharmaceutically acceptable salts thereof obtained by the process for preparing the compounds defined herein.

[0025] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Methods and materials are described herein for use in the present disclosure; other suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and are not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

[0026] Other features and advantages of the present disclosure will be apparent from the following detailed description, and from the claims. DETAILED DESCRIPTION OF THE INVENTION

[0027] definition The term "compound," as used herein, is meant to include all stereoisomers, geometric isomers, tautomers, and isotopically enriched variants of the depicted structure. A compound identified herein by name or structure as one particular tautomeric form is intended to include other tautomeric forms unless otherwise specified.

[0028] The term "tautomer," as used herein, refers to compounds that differ significantly in structure in the arrangement of atoms, but exist in easy and rapid equilibrium; the compounds provided herein may be represented as different tautomers; it should be understood that, where a compound has tautomeric forms, all tautomeric forms are intended to be within the scope of the present disclosure, and the naming of a compound does not exclude any tautomer. Examples of tautomeric forms include the following: [ka]

[0029] It will be understood that certain compounds provided herein may contain one or more asymmetric centers and, therefore, may be prepared and isolated in mixtures of isomers, such as racemic mixtures, or in enantiomerically pure form.

[0030] The term "halo" refers to a halogen, one of Group 17 of the periodic table. Specifically, the term refers to fluorine, chlorine, bromine, and iodine. Preferably, the term refers to fluorine or chlorine.

[0031] The term "C1-C6 alkyl" refers to a straight or branched hydrocarbon chain containing 1, 2, 3, 4, 5, or 6 carbon atoms, e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, and n-hexyl. An alkyl group can be unsubstituted or substituted with one or more substituents as described herein.

[0032] The term "C1-C6 haloalkyl" refers to a hydrocarbon chain substituted with at least one independently selected halogen atom at each occurrence, such as fluorine, chlorine, bromine, and iodine. The halogen atom may be present at any position on the hydrocarbon chain. For example, C1-C6 haloalkyl can refer to chloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl, chloroethyl, such as 1-chloroethyl and 2-chloroethyl, trichloroethyl, such as 1,2,2-trichloroethyl, 2,2,2-trichloroethyl, fluoroethyl, such as 1-fluoromethyl and 2-fluoroethyl, trifluoroethyl, such as 1,2,2-trifluoroethyl and 2,2,2-trifluoroethyl, chloropropyl, trichloropropyl, fluoropropyl, and trifluoropropyl.

[0033] The term "C1-C6 alkoxy" refers to a C1-C6 alkyl group attached to a molecule through an oxygen. It includes moieties where the alkyl portion can be straight or branched, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, and n-hexoxy.

[0034] As used herein, the term "hydroxyl" refers to the --OH radical.

[0035] The term "C1-C6 hydroxyalkyl" refers to a hydrocarbon chain substituted with one hydroxyl radical. The hydroxyl radical may be located at any position on the hydrocarbon chain. For example, C1-C6 hydroxyalkyl can refer to hydroxymethyl, hydroxyethyl, such as 1-hydroxyethyl and 2-hydroxyethyl, and 2-hydroxyisopropyl.

[0036] As used herein, the term "aryl" refers to a 6-10 total carbon monocyclic or bicyclic aromatic ring system. Non-limiting examples of aryl groups include phenyl and naphthyl.

[0037] As used herein, the term "heteroaryl" refers to a 5- to 10-membered monocyclic or bicyclic group in which each ring in the system is aromatic and in which one or more carbon atoms in at least one ring in the system is replaced by a heteroatom independently selected from N, O, and S. Non-limiting examples of heteroaryl groups include pyridine, pyrimidine, pyrrole, imidazole, and indole.

[0038] As used herein, the term "cycloalkyl" refers to a monocyclic or bicyclic carbon group having 3 to 10 ring atoms that is saturated or partially unsaturated; bicyclic systems include fused, spiro (optionally referred to as "spirocycloalkyl" groups), and bridged ring systems. Fused cycloalkyl groups can contain one ring that is aromatic and another ring that is saturated or partially saturated, such as 1,2,3,4-tetrahydronaphthalene and 2,3-dihydro-1H-indene. Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclohexyl, spiro[2.3]hexyl, and bicyclo[1.1.1]pentyl. As a substituent, for example, a cycloalkyl group on an akyl group can share a carbon atom with the alkyl chain.

[0039] The term "heterocyclyl" refers to a non-fully aromatic, saturated or partially unsaturated hydrocarbon monocyclic or bicyclic ring system having at least one heteroatom in the ring selected from N, O, and S. Bicyclic heterocyclyl groups include fused, spiro (optionally referred to as "spiroheterocyclyl" groups), and bridged ring systems. Fused heterocyclyl groups can contain one ring that is aromatic and another ring that is saturated or partially saturated, such as 5,6,7,8-tetrahydroquinoline and indoline. Heterocyclyl groups may also be referred to as "3- to 10-membered heterocyclyl groups," which are ring systems containing 3, 4, 5, 6, 7, 8, 9, or 10 atoms, with at least one atom being a heteroatom. For example, there may be 1, 2, or 3 heteroatoms, optionally 1 or 2. Heterocyclyl groups can be attached to the remainder of the molecule through any carbon atom or heteroatom, such as nitrogen. Exemplary heterocyclyl groups include, but are not limited to, piperidinyl, piperazinyl, morpholino, tetrahydropyranyl, azetidinyl, oxetanyl, and 2-azaspiro[3.3]heptanyl. As a substituent, for example, on an akyl group, a heterocyclyl group can share a carbon atom with the alkyl chain.

[0040] As used herein, the term "geminal" refers to substituent atoms or groups that are attached to the same atom in a molecule.

[0041] As used herein, the term "oxo" refers to an "=O" group attached to a carbon atom.

[0042] As used herein, the symbols [ka] indicates the point of attachment of the atom or moiety to the indicated atom or group in the remainder of the molecule.

[0043] It is to be understood that the A ring in compounds of formula (I) containing does not contain two adjacent oxygen atoms or two adjacent S atoms.

[0044] The compounds of formula (I) include pharmaceutically acceptable salts thereof. In addition, the compounds of formula (I) also include other salts of such compounds that are not necessarily pharmaceutically acceptable salts but may be useful as intermediates for preparing and / or purifying the compounds of formula (I) and / or for separating the enantiomers of the compounds of formula (I). Non-limiting examples of pharmaceutically acceptable salts of the compounds of formula (I) include trifluoroacetic acid and hydrochloride salts.

[0045] It will further be understood that the compounds of formula (I) or their salts may be isolated in the form of a solvate, and therefore any such solvates are included within the scope of the present disclosure. For example, the compounds of formula (I) and their salts may exist in unsolvated and solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like.

[0046] In some embodiments, compounds of Formula (I) include the compounds of Examples 1-96 and pharmaceutically acceptable salts and solvates thereof. In some embodiments, compounds of Formula (I) are in free base form. In some embodiments, compounds of Formula (I) are in salt form (e.g., pharmaceutically acceptable salts).

[0047] In some embodiments, the compound of Formula (I) includes its stereoisomers, as well as pharmaceutically acceptable salts and solvates. In some embodiments, the compound of Formula (I) is in free base form. In some embodiments, the compound of Formula (I) is in salt form.

[0048] The term "pharmaceutically acceptable" indicates that a compound, or salt thereof, or composition is chemically and / or toxicologically compatible with other ingredients comprising the formulation and / or the subject being treated therewith.

[0049] Protecting groups may be temporary substituents that protect potentially reactive functional groups from undesired chemical transformations. The selection of the specific protecting group to be used is well within the skill of one of ordinary skill in the art. Several considerations can determine the selection of a protecting group, including, but not limited to, the functional group to be protected, other functionalities present in the molecule, reaction conditions at each step of the synthetic sequence, other protecting groups present in the molecule, functional group resistance to conditions required to remove the protecting group, and reaction conditions for thermal decomposition of the compounds provided herein. The field of protecting group chemistry has been reviewed (Greene, TW; Wuts, PGMP Protective Groups in Organic Synthesis, 2nd ed.; Wiley: New York, 1991).

[0050] A nitrogen protecting group can be any temporary substituent that protects the amine moiety from undesired chemical transformations. Examples of the moiety formed when such a protecting group is attached to an amine include, but are not limited to, allylamine, benzylamine (e.g., benzylamine, p-methoxybenzylamine, 2,4-dimethoxybenzylamine, and tritylamine), acetylamide, trichloroacetamide, trifluoroacetamide, pent-4-enamide, phthalimide, carbamate (e.g., methylcarbamate, t-butylcarbamate, benzylcarbamate, allylcarbamate, 2,2,2-trichloroethylcarbamate, and 9-fluorenylmethylcarbamate), imine, and sulfonamide (e.g., benzenesulfonamide, p-toluenesulfonamide, and p-nitrobenzenesulfonamide).

[0051] An oxygen protecting group can be any temporary substituent that protects a hydroxyl moiety from undesired chemical transformations. Examples of moieties formed when such a protecting group is attached to a hydroxyl include, but are not limited to, esters (e.g., acetyl, t-butylcarbonyl, and benzoyl), benzyls (e.g., benzyl, p-methoxybenzyl, and 2,4-dimethoxybenzyl, and trityl), carbonates (e.g., methyl carbonate, allyl carbonate, 2,2,2-trichloroethyl carbonate, and benzyl carbonate), ketals, acetals, and ethers.

[0052] The compounds provided herein may also contain unnatural proportions of atomic isotopes in one or more of the atoms that constitute such compounds. That is, when an atom is specifically referred to in connection with a compound according to formula (I), it includes all isotopes and isotopic mixtures of that atom, whether naturally occurring or synthetically produced, in either natural abundance or isotopically enriched form. For example, unless otherwise specified, when hydrogen is referred to, the atom is 1 H, 2 H, 3 H, or mixtures thereof, and when carbon is mentioned, the atom is 11 C. 12 C. 13 C. 14 C, or mixtures thereof, and when nitrogen is mentioned, the atom 13 N, 14 N, 15 N, or mixtures thereof, and when oxygen is mentioned, the atom is 14 O. 15 O. 16 O. 17 O. 18 O, or mixtures thereof, and when fluoro is mentioned, the atom is 18 F, 19 F, or mixtures thereof. For example, in deuterated alkyl and deuterated alkoxy groups, one or more hydrogen atoms are replaced with deuterium ( 2Some of the aforementioned isotopes are radioactive, and therefore the compounds provided herein also include compounds with one or more isotopes of one or more atoms, including radioactive compounds, and mixtures thereof, in which one or more non-radioactive atoms are replaced by one of their radioactively enriched isotopes. Radiolabeled compounds are useful as therapeutic agents, e.g., cancer therapeutic agents, research reagents, e.g., assay reagents, and diagnostic agents, e.g., in vivo imaging agents. All isotopic variations of the compounds provided herein, whether radioactive or not, are intended to be encompassed within the scope of the present disclosure.

[0053] For illustrative purposes, general methods for preparing compounds and key intermediates are provided herein. For more detailed descriptions of the individual reaction steps, see the Examples section below. Those skilled in the art will appreciate that other synthetic routes may be used to synthesize the compounds of the invention. While specific starting materials and reagents are shown in the schemes and discussed below, other starting materials and reagents can be readily substituted to provide a variety of derivatives and / or reaction conditions. In addition, many of the compounds prepared by the methods described below can be further modified in light of this disclosure using conventional chemistry well known to those skilled in the art.

[0054] The ability of a test compound to act as a CDC7 inhibitor can be demonstrated by the biological and computational assays described herein. IC 50 The values ​​are shown in Tables 2 and 3.

[0055] The compounds of formula (I) (e.g., any one of formulas (IA)-(IP)), or pharmaceutically acceptable salts thereof, are useful for treating diseases and disorders that can be treated by CDC7 kinase inhibitors, such as CDC7-associated cancers, including hematological cancers and solid tumors.

[0056] As used herein, the term "treat" or "treatment" refers to curative or palliative measures. Beneficial or desired clinical results include, but are not limited to, the total or partial alleviation of symptoms associated with a disease or disorder or condition, whether detectable or undetectable, a decrease in the extent of the disease, a stabilization of the disease state (i.e., does not worsen), a delay or slowing of disease progression, an improvement or alleviation of the disease state (e.g., one or more symptoms of the disease), and remission (whether partial or complete). "Treatment" can also mean prolonging survival as compared to expected survival if not receiving treatment.

[0057] As used herein, the term "subject" refers to any animal, including mammals such as mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses, primates, and humans. In some embodiments, the subject is a human. In some embodiments, the subject is experiencing and / or exhibiting at least one symptom of the disease or disorder being treated and / or prevented.

[0058] In some embodiments, the subject has been identified or diagnosed with a cancer involving dysregulation of the CDC7 gene, CDC7 protein, or expression or activity or levels of any of them (CDC7-associated cancer) (e.g., as determined using a regulatory agency-approved, e.g., FDA-approved, assay or kit). In some embodiments, the subject has a tumor that is positive for dysregulation of the CDC7 gene, CDC7 protein, or expression or activity or levels of any of them (e.g., as determined using a regulatory agency-approved, e.g., FDA-approved, assay or kit). The subject may be a subject whose tumor is positive for dysregulation of the CDC7 gene, CDC7 protein, or expression or activity or levels of any of them (e.g., identified as positive using a regulatory agency-approved, e.g., FDA-approved, assay or kit). The subject may be a subject whose tumor has dysregulation of the CDC7 gene, CDC7 protein, or expression or activity or levels thereof (e.g., the tumor is identified as such using a regulatory agency-approved, e.g., FDA-approved, kit or assay). In some embodiments, the subject is suspected of having a CDC7-associated cancer. In some embodiments, the subject has clinical records indicating that the subject has a tumor with dysregulated expression or activity or levels of the CDC7 gene, CDC7 protein, or any of them (and optionally, the clinical records indicate that the subject should be treated with any of the compositions provided herein). In some embodiments, the subject is a pediatric subject. In some embodiments, the subject has been identified or diagnosed with a cancer determined to be associated with dysregulated expression or activity or levels of the CDC7 gene, CDC7 protein, or any of them (a CDC7-associated cancer) based on histological examination.

[0059] The term "pediatric subject," as used herein, refers to a subject under the age of 21 at the time of diagnosis or treatment. The term "pediatric" can be further divided into various subpopulations, including neonates (birth to 1 month), infants (1 month to 2 years), children (2 to 12 years), and adolescents (12 to 21 years, including but not limited to, their 22nd birthday). Berhman RE, Kliegman R, Arvin AM, Nelson WE. Nelson Textbook of Pediatrics, 15th Ed. Philadelphia: WB Saunders Company, 1996; Rudolph AM, et al. Rudolph's Pediatrics, 21st Ed. New York: McGraw-Hill, 2002; and Avery MD, First LR. Pediatric Medicine, 2nd Ed. Baltimore: Williams & Wilkins; 1994. In some embodiments, the pediatric subject is between birth and 28 days old, between 29 days old and under 2 years old, between 2 years old and under 12 years old, or between 12 years old and 21 years old (including but not including the 22nd birthday). In some embodiments, the pediatric subject is between birth and 28 days old, between 29 days old and under 1 year old, between 1 month old and under 4 months old, between 3 months old and under 7 months old, between 6 months old and under 1 year old, between 1 year old and under 2 years old, between 2 years old and under 3 years old, between 2 years old and under 7 years old, between 3 years old and under 5 years old, between 5 years old and under 10 years old, between 6 years old and under 13 years old, between 10 years old and under 15 years old, or between 15 years old and under 22 years old.

[0060] In some embodiments, the compounds of Formula (I) or pharmaceutically acceptable salts thereof are useful for preventing diseases and disorders defined herein (e.g., autoimmune diseases, inflammatory diseases, and cancer). The term "preventing," as used herein, means preventing the onset, recurrence, or spread, in whole or in part, of a disease or condition described herein, or a symptom thereof.

[0061] The term "CDC7-associated cancer," as used herein, refers to a cancer associated with or having dysregulation of the expression, activity, or levels of the CDC7 gene, CDC7 kinase (also referred to herein as the CDC7 kinase protein), or any (e.g., one or more) thereof (e.g., any of the types of dysregulation of the expression, activity, or levels of the CDC7 gene, CDC7 kinase, CDC7 kinase domain, or any of them described herein). Non-limiting examples of CDC7-associated diseases or disorders include, for example, cancer and gastrointestinal disorders such as irritable bowel syndrome (IBS).

[0062] The term "CDC7-associated cancer," as used herein, refers to a cancer associated with or having dysregulation of the CDC7 gene, CDC7 kinase, or the expression or activity or levels of any of them. Non-limiting examples of CDC7-associated cancers are described herein.

[0063] The phrase "dysregulated expression or activity or levels of the CDC7 gene, CDC7 kinase, or any of them" refers to a genetic mutation (e.g., a chromosomal translocation resulting in expression of a fusion protein comprising the CDC7 kinase domain and a fusion partner, a mutation in the CDC7 gene resulting in expression of a CDC7 protein with at least one amino acid deletion compared to the wild-type CDC7 protein, a mutation in the CDC7 gene resulting in expression of a CDC7 protein with one or more point mutations compared to the wild-type CDC7 protein, a mutation in the CDC7 gene resulting in expression of a CDC7 protein with at least one inserted amino acid compared to the wild-type CDC7 protein, a gene duplication resulting in increased levels of the CDC7 protein in a cell, or a mutation in a regulatory sequence (e.g., a promoter and / or enhancer) resulting in increased levels of the CDC7 protein in a cell), an alternatively spliced ​​version of the CDC7 mRNA resulting in a CDC7 protein with at least one amino acid deletion in the CDC7 protein compared to the wild-type CDC7 protein, or increased expression (e.g., increased levels) of wild-type CDC7 kinase in a mammalian cell due to aberrant cell signaling and / or dysregulated autocrine / paracrine signaling (e.g., compared to control non-cancerous cells). As another example, dysregulation of the expression, activity, or levels of the CDC7 gene, CDC7 protein, or any of them may be a mutation in the CDC7 gene that encodes a CDC7 protein that is constitutively active or has increased activity compared to the protein encoded by a CDC7 gene that does not contain the mutation. As a further example, an increase in the copy number of the CDC7 gene may result in overexpression of the CDC7 kinase. For example, dysregulation of the expression, activity, or levels of the CDC7 gene, CDC7 protein, or any of them may be the result of a genetic or chromosomal translocation that results in the expression of a fusion protein containing a first portion of CDC7 that includes a functional kinase domain and a second portion of a partner protein (i.e., that is not CDC7).In some instances, dysregulation of the CDC7 gene, the CDC7 protein, or the expression or activity or levels of any of them may be the result of a genetic translocation between one CDC7 gene and another non-CDC7 gene.

[0064] The term "wild-type" refers to a nucleic acid (e.g., a CDC7 gene or CDC7 mRNA) or protein (e.g., a CDC7 protein) found in a subject who does not have (and optionally does not have an elevated risk of developing and / or is not suspected of having) a CDC7-associated disease, e.g., a CDC7-associated cancer, or found in a cell or tissue derived from a subject who does not have (and optionally does not have an elevated risk of developing and / or is not suspected of having) a CDC7-associated disease, e.g., a CDC7-associated cancer.

[0065] The term "regulatory agency" refers to a national agency that approves pharmaceutical agents for medical use in a country. For example, a non-limiting example of a regulatory agency is the U.S. Food and Drug Administration (FDA).

[0066] As used herein, compounds of formula (I): [ka] or a pharmaceutically acceptable salt thereof, wherein: R 1 is hydrogen or halogen, R 2 is a 5- to 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from the group consisting of halogen and C1-C6 alkyl, or is a 5- to 6-membered heterocyclyl optionally substituted with 1 or 2 substituents independently selected from the group consisting of halogen and C1-C6 alkyl; R 3 is hydrogen or C1-C6 alkyl, Ring A is C6-C10 cycloalkyl or 6- to 10-membered heterocyclyl; Each R4 is halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, C1-C6 haloalkyl, C1-C6 alkoxy(C1-C6 alkyl)-, -C(=O)C1-C6 alkyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -NR A R B and 4- to 6-membered heterocyclyl optionally substituted with 1 or 2 independently selected halogens; A and R B are independently hydrogen or C1-C6 alkyl; m is 0, 1, 2, 3, or 4.

[0067] In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, m is 3. In some embodiments, m is 4.

[0068] In some embodiments, R 2 is a 5-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from the group consisting of halogen and C1-C6 alkyl. In some embodiments, R 2 is a 5-membered heteroaryl substituted with 1 or 2 substituents independently selected from the group consisting of halogen and C1-C6 alkyl.

[0069] In some embodiments, R 2 is a 5-membered heteroaryl group selected from the group consisting of pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, thiophenyl, oxazolyl, isoxazolyl, isothiazolyl, thiazolyl, fluzanyl, oxadiazolyl, thiadiazolyl, oxatriazolyl, and thiatriazolyl. In some embodiments, R 2 The 5-membered heteroaryl of R is pyrazolyl or isothiazolyl. 2 is 4-pyrazolyl, 5-pyrazolyl, or 5-isothiazolyl.

[0070] In some embodiments, R 2 is a 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from the group consisting of halogen and C1-C6 alkyl. In some embodiments, R 2 is a 6-membered heteroaryl substituted with 1 or 2 substituents independently selected from the group consisting of halogen and C1-C6 alkyl.

[0071] In some embodiments, R 2 is a 6-membered heteroaryl group selected from the group consisting of pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, and triazinyl. 2 The 6-membered heteroaryl of R is pyridyl, pyrimidinyl, or pyridazinyl. 2 In some embodiments, the 6-membered heteroaryl of R is pyridyl. 2 is 4-pyridyl. In some embodiments, R 2 is 4-pyrimidinyl. In some embodiments, R 2 is 4-pyridazinyl.

[0072] In some embodiments, R 2 The heteroaryl in R is substituted with one substituent selected from halogen and C1-C6 alkyl. 2 The heteroaryl in R is substituted with one substituent selected from fluoro, chloro, and methyl. 2 The heteroaryl in R is substituted with two substituents independently selected from halogen and C1-C6 alkyl. 2 The heteroaryl of is substituted with two substituents independently selected from fluoro and methyl.

[0073] In some embodiments, R 2 The above substituents are the same. In some embodiments, R 2 The above substituents are different.

[0074] In some embodiments, R 2 is an unsubstituted 5-6 membered heteroaryl such as unsubstituted pyrazolyl, isothiazolyl, pyridyl, or pyridazinyl.

[0075] In some embodiments, R 2 is a 5- to 6-membered heterocyclyl optionally substituted with 1 or 2 substituents independently selected from the group consisting of halogen and C1-C6 alkyl. In some embodiments, R 2 is unsubstituted 5-6 membered heterocyclyl. In some embodiments, R 2 is morpholinyl, piperidinyl, or piperazinyl. In some embodiments, R 2 is morpholinyl. In some embodiments, R 2 is 4-morpholinyl.

[0076] In some embodiments, Ring A is C6-C10 cycloalkyl. In some embodiments, Ring A is cyclohexyl. In some embodiments, Ring A is bicyclo[2.2.2]octanyl.

[0077] In some embodiments, ring A is 6-10 membered heterocyclyl. In some embodiments, ring A is tetrahydropyranyl. In some embodiments, ring A is piperidinyl.

[0078] In some embodiments, one or two R 4 is independently halogen. In some embodiments, one R 4 is fluoro or chloro. In some embodiments, one R 4 is fluoro. In some embodiments, one R 4 is chloro. In some embodiments, two R 4 is fluoro.

[0079] In some embodiments, one R 4is a hydroxyl.

[0080] In some embodiments, one R 4 is independently C-C alkyl. In some embodiments, one or two R 4 is an independently selected C-C alkyl. In some embodiments, one R 4 are independently selected from the group consisting of -CH3, -CH2CH3, -CH(CH3)2, -CH(CH3)CH2CH3, -C(CH3)2CH2CH3, and -C(CH3)3.

[0081] In some embodiments, each R 4 is independently C-C alkoxy. In some embodiments, one R 4 is C1-C6 alkoxy. In some embodiments, each R 4 is independently methoxy, ethoxy, or isopropoxy. In some embodiments, one R 4 is methoxy, ethoxy, or isopropoxy. In some embodiments, each R 4 is methoxy. In some embodiments, one R 4 is methoxy.

[0082] In some embodiments, one R 4 is C1-C6 hydroxyalkyl. In some embodiments, one R 4 is C1-C6 hydroxyalkyl. In some embodiments, one R 4 is —CHOH or —C(CH)OH. In some embodiments, one R 4 is —CH2OH or —C(CH3)2OH.

[0083] In some embodiments, one R 4 is C1-C6 haloalkyl. In some embodiments, one or two R 4 is an independently selected C1-C6 haloalkyl. In some embodiments, one R 4is -CF3. In some embodiments, one R 4 is -CHF. In some embodiments, one R 4 is -CF3. In some embodiments, one R 4 is -CHF. In some embodiments, one R 4 is -CF2CH3. In some embodiments, one R 4 is CH2CF2CH3.

[0084] In some embodiments, one R 4 is C-C alkoxy(C-C alkyl)-. In some embodiments, one R 4 is C1-C3 alkoxy(C1-C3 alkyl)-. In some embodiments, one R 4 is independently methoxy(C-C alkyl)-. In some embodiments, one R 4 is -CH2OCH3.

[0085] In some embodiments, one R 4 is —C(═O)C1-C6 alkyl. In some embodiments, R 4 is C(=O)CH3.

[0086] In some embodiments, one R 4 is C2-C6 alkynyl. In some embodiments, one R 4 is 1-propynyl.

[0087] In some embodiments, one R 4 is C-C cycloalkyl. In some embodiments, R 4 is cyclopentyl.

[0088] In some embodiments, one R 4 is -NR A R B In some embodiments, R A and R Bis hydrogen and R A and R B and the other is C1-C6 alkyl. In some embodiments, R A and R B are both C1-C6 alkyl. In some embodiments, R 4 is -NH2.

[0089] In some embodiments, one R 4 is a 4- to 6-membered heterocyclyl optionally substituted with 1 or 2 independently selected halogens. In some embodiments, one R 4 is a 4- to 6-membered heterocyclyl substituted with 1 or 2 independently selected halogens. 4 is a 4-6 membered heterocyclyl substituted with 1 or 2 fluoro. In some embodiments, one R 4 is a 4- to 6-membered heterocyclyl substituted with a germinal difluoro group.

[0090] In some embodiments, one R 4 is azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuryl, piperidinyl, piperazinyl, tetrahydrofuryl, mofolinyl, or dioxanyl, each optionally substituted with one or two independently selected halogens. 4 is pyrrolidinyl optionally substituted with one or two independently selected halogens. In some embodiments, one R 4 is pyrrolidinyl substituted with one or two independently selected halogens. In some embodiments, one R 4 is azetidinyl optionally substituted with one or two independently selected halogens. In some embodiments, one R 4is azetidinyl substituted with one or two independently selected halogens. In some embodiments, one or two independently selected halogens are fluoro. In some embodiments, two independently selected halogens are germinal difluoro groups.

[0091] In some embodiments, one R 4 is an unsubstituted 4-6 membered heterocyclyl. In some embodiments, R 4 The unsubstituted 4-6 membered heterocyclyl is azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuryl, piperidinyl, piperazinyl, tetrahydrofuryl, mofolinyl, or dioxanyl. 4 is unsubstituted pyrrolidinyl or unsubstituted azetidinyl.

[0092] In some embodiments, one R 4 teeth, [ka] is selected from the group consisting of:

[0093] In some embodiments, m is 2, 3, or 4, and 2, 3, or 4 R 4 In some embodiments, m is 2 and both (i.e., two) R 4 In some embodiments, m is 3 and three R 4 In some embodiments, m is 4 and R 4 Each pair of groups is geminal.

[0094] In some embodiments, R 1 is halogen. In some embodiments, R 1 is fluoro or chloro. In some embodiments, R 1 is fluoro. In some embodiments, R 1is hydrogen.

[0095] In some embodiments, R 3 is C1-C6 alkyl. In some embodiments, R 3 is methyl. In some embodiments, R 3 is hydrogen.

[0096] In some embodiments, m is 2 and one R 4 is hydroxyl, and the other R 4 is C1-C6 alkyl.

[0097] In some embodiments, m is 2 and each R 4 is fluoro.

[0098] In some embodiments, m is 2 and one R 4 is hydroxyl, and the other R 4 is C1-C6 haloalkyl.

[0099] In some embodiments, m is 2 and one R 4 is hydroxyl, and the other R 4 is C2-C6 alkynyl.

[0100] In some embodiments, m is 2 and one R 4 is hydroxyl, and the other R 4 is C3-C6 cycloalkyl.

[0101] In some embodiments, m is 2 and one R 4 is hydroxyl, and the other R 4 is C1-C6 alkoxy(C1-C6 alkyl)-.

[0102] In some embodiments, m is 2 and each R 4 is an independently selected C1-C6 alkyl.

[0103] In some embodiments, m is 2 and one R 4 is C1-C6 alkyl, and the other R 4 is C1-C6 hydroxyalkyl.

[0104] In some embodiments, m is 2 and one R 4 is C1-C6 alkyl, and the other R 4 is C1-C6 alkoxy.

[0105] In some embodiments, m is 2 and one R 4 is C1-C6 alkyl, and the other R 4 is -NR A R B is.

[0106] In some embodiments, m is 2 and one R 4 is C1-C6 alkyl, and the other R 4 is a 4- to 6-membered heterocyclyl optionally substituted with 1 or 2 independently selected halogens.

[0107] In some embodiments, m is 4 and two R 4 is fluoro and one R 4 is C1-C6 alkoxy, and one R 4 is C1-C6 alkyl.

[0108] In some embodiments, m is 4 and two R 4 is fluoro and one R 4 is C1-C6 hydroxyalkyl, and one R 4 is C1-C6 alkyl.

[0109] In some embodiments, m is 4 and two R 4 is fluoro and one R 4 is a hydroxyl and one R 4 is C1-C6 alkyl.

[0110] In some embodiments, m is 4 and two R 4 is fluoro and one R 4 is a hydroxyl and one R 4 is C1-C6 alkoxy(C1-C6 alkyl)-.

[0111] In some embodiments, R 1 is hydrogen and R 2 is unsubstituted pyrazolyl, m is 2, and one R 4 is hydroxyl, and the other R 4 is C1-C6 alkyl.

[0112] In some embodiments, R 1 is hydrogen and R 2 is unsubstituted pyrazolyl, m is 2, and each R 4 is fluoro.

[0113] In some embodiments, R 1 is hydrogen and R 2 is unsubstituted pyrazolyl, m is 2, and one R 4 is hydroxyl, and the other R 4 is C1-C6 haloalkyl.

[0114] In some embodiments, R 1 is hydrogen and R 2 is unsubstituted pyrazolyl, m is 4, and two R 4 is fluoro and one R 4 is C1-C6 alkoxy, and one R 4 is C1-C6 alkyl.

[0115] In some embodiments, R 1 is hydrogen and R 2 is unsubstituted pyrazolyl, m is 4, and two R 4 is fluoro and one R 4 is C1-C6 hydroxyalkyl, and one R 4is C1-C6 alkyl.

[0116] In some embodiments, R 1 is hydrogen and R 2 is unsubstituted pyridinyl, m is 2, and one of R 4 is hydroxyl, and the other R 4 is C1-C6 alkyl.

[0117] In some embodiments, R 1 is hydrogen and R 2 is unsubstituted pyridinyl, m is 2, and each R 4 is fluoro.

[0118] In some embodiments, R 1 is hydrogen and R 2 is unsubstituted pyridinyl, m is 2, and one of R 4 is hydroxyl, and the other R 4 is C1-C6 haloalkyl.

[0119] In some embodiments, R 1 is hydrogen and R 2 is unsubstituted pyridinyl, m is 4, and two R 4 is fluoro and one R 4 is C1-C6 alkoxy, and one R 4 is C1-C6 alkyl.

[0120] In some embodiments, R 1 is hydrogen and R 2 is unsubstituted pyridinyl, m is 4, and two R 4 is fluoro and one R 4 is C1-C6 hydroxyalkyl, and one R 4 is C1-C6 alkyl.

[0121] In some embodiments, R 1 is hydrogen and R 2is unsubstituted isothiazolyl, m is 2, and one R 4 is hydroxyl, and the other R 4 is C1-C6 alkyl.

[0122] In some embodiments, R 1 is hydrogen and R 2 is unsubstituted isothiazolyl, m is 2, and each R 4 is fluoro.

[0123] In some embodiments, R 1 is hydrogen and R 2 is unsubstituted isothiazolyl, m is 2, and one R 4 is hydroxyl, and the other R 4 is C1-C6 haloalkyl.

[0124] In some embodiments, R 1 is hydrogen and R 2 is unsubstituted isothiazolyl, m is 4, and two R 4 is fluoro and one R 4 is C1-C6 alkoxy, and one R 4 is C1-C6 alkyl.

[0125] In some embodiments, R 1 is hydrogen and R 2 is unsubstituted isothiazolyl, m is 4, and two R 4 is fluoro and one R 4 is C1-C6 hydroxyalkyl, and one R 4 is C1-C6 alkyl.

[0126] In some embodiments, ring A is cyclohexyl, m is 2, and one R 4 is hydroxyl, and the other R 4 is C1-C6 alkyl.

[0127] In some embodiments, ring A is cyclohexyl, m is 2, and each R 4 is fluoro.

[0128] In some embodiments, ring A is cyclohexyl, m is 2, and one R 4 is hydroxyl, and the other R 4 is C1-C6 haloalkyl.

[0129] In some embodiments, ring A is cyclohexyl, m is 4, and two R 4 is fluoro and one R 4 is C1-C6 alkoxy, and one R 4 is C1-C6 alkyl.

[0130] In some embodiments, ring A is cyclohexyl, m is 4, and two R 4 is fluoro and one R 4 is C1-C6 hydroxyalkyl, and one R 4 is C1-C6 alkyl.

[0131] In some embodiments, ring A is bicyclo[2.2.2]octanyl, m is 2, and one R 4 is hydroxyl, and the other R 4 is C1-C6 alkyl.

[0132] In some embodiments, ring A is bicyclo[2.2.2]octanyl, m is 2, and each R 4 is fluoro.

[0133] In some embodiments, ring A is bicyclo[2.2.2]octanyl, m is 2, and one R 4 is hydroxyl, and the other R 4 is C1-C6 haloalkyl.

[0134] In some embodiments, ring A is bicyclo[2.2.2]octanyl, m is 4, and two R 4 is fluoro and one R 4 is C1-C6 alkoxy, and one R 4 is C1-C6 alkyl.

[0135] In some embodiments, ring A is bicyclo[2.2.2]octanyl, m is 4, and two R 4 is fluoro and one R 4 is C1-C6 hydroxyalkyl, and one R 4 is C1-C6 alkyl.

[0136] In some embodiments, ring A is tetrahydropyranyl, m is 2, and one R 4 is hydroxyl, and the other R 4 is C1-C6 alkyl.

[0137] In some embodiments, ring A is tetrahydropyranyl, m is 2, and each R 4 is fluoro.

[0138] In some embodiments, ring A is tetrahydropyranyl, m is 2, and one R 4 is hydroxyl, and the other R 4 is C1-C6 haloalkyl.

[0139] In some embodiments, ring A is piperidinyl, m is 3, and one R 4 is a hydroxyl and one R 4 is C1-C6 alkyl and one R 4 is —C(═O)C1-C6 alkyl. In some embodiments, when ring A is piperidinyl, one R 4 is attached to the piperidinyl nitrogen atom.

[0140] In some embodiments, ring A is tetrahydropyranyl, m is 4, and two R 4 is fluoro and one R 4 is C1-C6 alkoxy, and one R 4 is C1-C6 alkyl.

[0141] In some embodiments, ring A is tetrahydropyranyl, m is 4, and two R 4 is fluoro and one R 4 is C1-C6 hydroxyalkyl, and one R 4 is C1-C6 alkyl.

[0142] In some embodiments, compounds of formula (IA), (IB), (IC), (ID), (IE), (IF), (IG), (IH), (II), (IJ), (IK), (IL), (IM), (IN), (IO), and (IP) described herein, and pharmaceutically acceptable salts thereof, R 1 , R 2 , R 4 and m is as described in relation to formula (I), or a pharmaceutically acceptable salt thereof.

[0143] In some embodiments, the compound of formula (I) is a compound of formula (IA): [ka] or a pharmaceutically acceptable salt thereof.

[0144] In some embodiments, the compound of formula (I) is a compound of formula (IB): [ka] or a pharmaceutically acceptable salt thereof.

[0145] In some embodiments, the compound of formula (I) is a compound of formula (IC): [ka] or a pharmaceutically acceptable salt thereof.

[0146] In some embodiments, the compound of formula (I) is a compound of formula (ID): [ka] or a pharmaceutically acceptable salt thereof.

[0147] In some embodiments, the compound of formula (I) is a compound of formula (IE): [ka] or a pharmaceutically acceptable salt thereof.

[0148] In some embodiments, the compound of formula (I) is a compound of formula IF): [ka] or a pharmaceutically acceptable salt thereof.

[0149] In some embodiments, the compound of formula (I) is a compound of formula IG): [ka] or a pharmaceutically acceptable salt thereof.

[0150] In some embodiments, the compound of formula (I) is a compound of formula IH): [ka] or a pharmaceutically acceptable salt thereof.

[0151] In some embodiments, the compound of formula (I) is a compound of formula II): [ka] or a pharmaceutically acceptable salt thereof.

[0152] In some embodiments, the compound of formula (I) is a compound of formula IJ): [ka] or a pharmaceutically acceptable salt thereof.

[0153] In some embodiments, the compound of formula (I) is a compound of formula IK): [ka] or a pharmaceutically acceptable salt thereof.

[0154] In some embodiments, the compound of formula (I) is a compound of formula IL): [ka] or a pharmaceutically acceptable salt thereof.

[0155] In some embodiments, the compound of formula (I) is a compound of formula IM): [ka] or a pharmaceutically acceptable salt thereof.

[0156] In some embodiments, the compound of formula (I) is a compound of formula IN): [ka] or a pharmaceutically acceptable salt thereof.

[0157] In some embodiments, the compound of formula (I) is a compound of formula IO): [ka] or a pharmaceutically acceptable salt thereof.

[0158] In some embodiments, the compound of formula (I) is a compound of formula IP): [ka] or a pharmaceutically acceptable salt thereof.

[0159] Table 1 shows compounds of formula (I). Unless otherwise specified, all stereochemistry in Table 1 is understood to be arbitrarily assigned.

[0160] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is a compound selected from Table 1, or a pharmaceutically acceptable salt thereof. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11] [Table 1-12] [Table 1-13]

[0161] Treatment method Provided herein are methods for treating cancer (e.g., a CDC7-associated cancer) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. For example, provided herein are methods for treating a CDC7-associated cancer in a subject in need thereof, comprising: a) detecting dysregulation of the expression, activity, or level of the CDC7 gene, CDC7 kinase, or any of them in a sample from the subject; and b) administering a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof. In some embodiments, the dysregulation of the expression, activity, or level of the CDC7 gene, CDC7 kinase, or any of them comprises one or more fusion proteins.

[0162] In some embodiments of any of the methods or uses described herein, the cancer (e.g., a CDC7-associated cancer) is a hematological cancer. In some embodiments of any of the methods or uses described herein, the cancer (e.g., a CDC7-associated cancer) is a solid tumor. In some embodiments of any of the methods or uses described herein, the cancer (e.g., a CDC7-associated cancer) is selected from the group consisting of lung cancer (e.g., small cell lung cancer or non-small cell lung cancer), thyroid cancer (e.g., papillary thyroid cancer, medullary thyroid cancer (e.g., sporadic medullary thyroid cancer or hereditary medullary thyroid cancer), differentiated thyroid cancer, recurrent thyroid cancer, and refractory differentiated thyroid cancer), thyroid adenoma, endocrine neoplasm, lung adenocarcinoma, bronchiolopulmonary cell carcinoma, multiple endocrine neoplasia type 2A or 2B (MEN2A or MEN2B, respectively), pheochromocytoma, parathyroid hyperplasia, breast cancer,carcinoma), breast tumor, colorectal cancer (e.g., metastatic colorectal cancer), papillary renal cell carcinoma, ganglioneuromatosis of the gastrointestinal mucosa, inflammatory myofibroblastic tumor, or cervical cancer. In some embodiments of any of the methods or uses described herein, the cancer (e.g., a CDC7-associated cancer) is selected from the group consisting of acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), cancer in adolescents, adrenocortical carcinoma, anal cancer, appendix cancer, astrocytoma, atypical teratoma / rhabdoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain stem glioma, brain tumor, breast cancer, bronchial tumor, Burkitt's lymphoma, carcinoma-like tumor, cancer of unknown primary, cardiac tumor, cervical cancer, childhood cancer, chordoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myeloproliferative neoplasm, site-specific neoplasm, neoplasm, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, cutaneous angiosarcoma, cholangiocarcinoma, ductal carcinoma in situ, embryonal tumor, endometrial cancer, ependymoma, esophageal cancer, nasal nerve Blastoma, Ewing's sarcoma, extracranial germ cell tumor, extragonadal germ cell tumor, extrahepatic bile duct cancer, eye cancer, fallopian tube cancer, fibrous histiocytoma of bone, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic disease, glioma, hairy cell tumor, hairy cell leukemia, head and neck cancer, breast neoplasm, head and neck neoplasm, CNS tumor, primary CNS tumor, cardiac cancer, hepatocellular carcinoma, Histiocytosis, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, pancreatic islet cell tumor, pancreatic neuroendocrine tumor, Kaposi's sarcoma, kidney cancer, Langerhans cell histiocytosis, laryngeal cancer, leukemia, lip and oral cavity cancer, liver cancer, lung cancer, lymphoma, macroglobulinemia, malignant fibrous histiocytoma of bone, bone cancer, melanoma, Merkel cell carcinoma, mesothelioma, metastatic squamous cell carcinoma of the neck, midline duct carcinoma, mouth cancer cancer), multiple endocrine neoplasia syndrome, multiple myeloma, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative neoplasm, neoplasm by site, neoplasm, myeloid leukemia, myeloid leukemia, multiple myeloma, myeloproliferative neoplasm, nasal cavity and paranasal sinus cancer, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, lung neoplasm, lung cancer, respiratory tract neoplasm, bronchogenic carcinoma, bronchial neoplasm, oral cancercancer), oral cavity cancer, lip cancer, oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillomatosis, paraganglioma, paranasal sinus and nasal cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pituitary cancer, plasma cell neoplasms, pleuropulmonary blastoma, pregnancy-associated breast cancer, primary central nervous system lymphoma, primary peritoneal cancer, prostate cancer, rectal cancer, colon cancer, colon neoplasms, renal cell carcinoma, CDC7 retinoblastoma, thoracic stenosis, thyroid cancer ... The cancer is selected from the group consisting of rhabdomyosarcoma, salivary gland cancer, sarcoma, Sezary syndrome, skin cancer, Spitz tumor, small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, squamous cell cervical cancer, gastric cancer, T-cell lymphoma, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and uCDC7er, cancer of unknown primary site, uCDC7hral cancer, urethral cancer, uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, and Wilms' tumor.

[0163] In some embodiments, the hematological cancer (e.g., a hematological cancer that is a CDC7-associated cancer) is a leukemia, lymphoma (non-Hodgkin's lymphoma), Hodgkin's disease (also called Hodgkin's lymphoma), and myeloma, such as acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myelomonocytic leukemia (CMML), and chronic myelomonocytic leukemia (CML). ), chronic neutrophilic leukemia (CNL), acute anaplastic leukemia (AUL), anaplastic large cell lymphoma (ALCL), prolymphocytic leukemia (PML), juvenile myelomonocytic leukemia (JMML), adult T-cell ALL, AML with trilineage myelodysplasia (AML / TMDS), mixed lineage leukemia (MLL), myelodysplastic syndromes (MDS), myeloproliferative disorders (MPDs), and multiple myeloma (MM). Additional examples of hematological cancers include myeloproliferative disorders (MPDs), such as polycythemia vera (PV), essential thrombocytopenia (ET), and idiopathic primary myelofibrosis (IMF / IPF / PMF). In some embodiments, the hematological cancer (e.g., a hematological cancer that is a CDC7-associated cancer) is AML or CMML.

[0164] In some embodiments, the cancer (e.g., a CDC7-associated cancer) is a solid tumor. Examples of solid tumors (e.g., solid tumors that are CDC7-associated cancers) include, for example, thyroid cancer (e.g., papillary thyroid cancer, medullary thyroid cancer), lung cancer (e.g., lung adenocarcinoma, small cell lung cancer), pancreatic cancer, pancreatic ductal carcinoma, breast cancer, colon cancer, colorectal cancer, prostate cancer, renal cell carcinoma, head and neck tumors, neuroblastoma, and melanoma. See, for example, Nature Reviews Cancer, 2014, 14, 173-186.

[0165] In some embodiments, the cancer is selected from the group consisting of lung cancer, papillary thyroid cancer, medullary thyroid cancer, differentiated thyroid cancer, recurrent thyroid cancer, refractory differentiated thyroid cancer, multiple endocrine neoplasia type 2A or type 2B (MEN2A or MEN2B, respectively), pheochromocytoma, parathyroid hyperplasia, breast cancer, colorectal cancer, papillary renal cell carcinoma, ganglioneuromatosis of the gastrointestinal mucosa, and cervical cancer.

[0166] In some embodiments, the subject is a human.

[0167] The compounds of formula (I) and their pharmaceutically acceptable salts and solvates are also useful in treating CDC7-associated cancers.

[0168] Accordingly, also provided herein are methods for treating a subject diagnosed or identified as having a CDC7-associated cancer, e.g., any of the exemplary CDC7-associated cancers disclosed herein, comprising administering to the subject a therapeutically effective amount of a compound of Formula (I) as defined herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is selected from Examples 1-96, or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, administered to the subject is a compound of Formula (IA), (IB), (IC), (ID), (IE), (IF), (IG), (IH), (II), (IJ), (IK), (IL), (IM), (IN), (IO), or (IP), or a pharmaceutically acceptable salt of any of the foregoing.

[0169] Dysregulation of the expression, activity, or levels of CDC7 kinase, the CDC7 gene, or any (e.g., one or more) thereof can affect tumorigenesis. For example, a fusion protein can have increased kinase activity compared to wild-type CDC7 protein, increased expression (e.g., increased levels) of wild-type CDC7 kinase in mammalian cells can result from aberrant cell signaling and / or dysregulated autocrine / paracrine signaling (e.g., compared to control non-cancerous cells), and CDC7 mRNA splice variants can also result in dysregulation of CDC7.

[0170] In some embodiments, compounds provided herein exhibit brain and / or central nervous system (CNS) penetrance. Such compounds are capable of crossing the blood-brain barrier and inhibiting CDC7 kinase in the brain and / or other CNS structures. In some embodiments, compounds provided herein are capable of crossing the blood-brain barrier in an effective amount. For example, treating a subject with cancer (e.g., a CDC7-associated cancer, such as a CDC7-associated brain or CNS cancer) can include administering (e.g., orally administering) a compound to the subject. In some such embodiments, compounds provided herein are useful for treating primary or metastatic brain tumors. For example, the compounds may be used to treat one or more gliomas, such as glioblastoma (also known as glioblastoma multiforme), astrocytoma, oligodendroglioma, ependymoma, and mixed glioma, meningioma, medulloblastoma, ganglioglioma, schwannoma (schwannoma), and craniopharyngioma (see, e.g., the tumors listed in Louis, DNet al. Acta Neuropathol 131(6), 803-820 (June 2016)). In some embodiments, the brain tumor is a primary brain tumor. In some embodiments, the subject has previously been treated with another anti-cancer agent, for example, another CDC7 inhibitor (e.g., a compound that is not a compound of general formula (I)), or a multi-kinase inhibitor. In some embodiments, the brain tumor is a metastatic brain tumor. In some embodiments, the subject has previously been treated with another anti-cancer agent, for example, another CDC7 inhibitor (e.g., a compound that is not a compound of formula (I)), or a multi-kinase inhibitor.

[0171] In some embodiments of any of the methods or uses described herein, the assay used to determine whether a subject has dysregulated expression, activity, or levels of the CDC7 gene, or CDC7 kinase, or any of them, using a sample from the subject can include, for example, next-generation sequencing, immunohistochemistry, fluorescence microscopy, break-apart FISH analysis, Southern blotting, Western blotting, FACS analysis, Northern blotting, and PCR-based amplification (e.g., RT-PCR and quantitative real-time RT-PCR). As is well known in the art, the assay is typically performed using, for example, at least one labeled nucleic acid probe or at least one labeled antibody or antigen-binding fragment thereof. The assay can utilize other detection methods known in the art to detect dysregulated expression, activity, or levels of the CDC7 gene, CDC7 kinase, or any of them. In some embodiments, the sample is a biological sample or biopsy sample (e.g., a paraffin-embedded biopsy sample) from the subject. In some embodiments, the subject is suspected of having a CDC7-associated cancer, has one or more symptoms of a CDC7-associated cancer, and / or is at high risk of developing a CDC7-associated cancer).

[0172] In some embodiments, dysregulation of the expression, activity, or level of the CDC7 gene, CDC7 kinase, or any of them can be identified using a liquid biopsy (variously referred to as a fluid biopsy or fluid-phase biopsy). Liquid biopsy methods can be used to detect total tumor burden and / or dysregulation of the expression, activity, or level of the CDC7 gene, CDC7 kinase, or any of them. Liquid biopsies can be performed on biological samples that are relatively easily obtained from a subject (e.g., via a simple blood draw) and are generally less invasive than traditional methods used to detect tumor burden and / or dysregulation of the expression, activity, or level of the CDC7 gene, CDC7 kinase, or any of them. In some embodiments, liquid biopsies can be used to detect the presence of dysregulation of the expression, activity, or level of the CDC7 gene, CDC7 kinase, or any of them at an earlier stage than traditional methods. In some embodiments, biological samples used in liquid biopsies can include blood, plasma, urine, cerebrospinal fluid, saliva, sputum, bronchoalveolar lavage fluid, bile, lymph, cyst fluid, stool, ascites, and combinations thereof. In some embodiments, liquid biopsy can be used to detect circulating tumor cells (CTCs). In some embodiments, liquid biopsy can be used to detect cell-free DNA. In some embodiments, the cell-free DNA detected using liquid biopsy is circulating tumor DNA (ctDNA) derived from tumor cells. Analysis of ctDNA (e.g., using highly sensitive detection techniques such as, but not limited to, next-generation sequencing (NGS), conventional PCR, digital PCR, or microarray analysis) can be used to identify dysregulation of the expression, activity, or levels of the CDC7 gene, CDC7 kinase, or any of them.

[0173] In some embodiments, ctDNA derived from a single gene can be detected using a liquid biopsy. In some embodiments, ctDNA derived from multiple genes (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 or more genes, or any number in between) can be detected using a liquid biopsy. In some embodiments, ctDNA derived from multiple genes can be detected using any of a variety of commercially available test panels (e.g., commercially available test panels designed to detect dysregulation of the expression, activity, or levels of the CDC7 gene, CDC7 kinase, or any of them). Liquid biopsies can be used to detect dysregulation of expression, activity, or levels of the CDC7 gene, CDC7 kinase, or any of them, including, but not limited to, point mutations or single nucleotide variants (SNVs), copy number variants (CNVs), gene fusions (e.g., translocations or rearrangements), insertions, deletions, or any combination thereof. In some embodiments, liquid biopsies can be used to detect germline mutations. In some embodiments, liquid biopsies can be used to detect somatic mutations. In some embodiments, liquid biopsies can be used to detect primary genetic mutations (e.g., primary mutations or primary fusions associated with early onset of a disease, e.g., cancer). In some embodiments, dysregulation of expression, activity, or levels of the CDC7 gene, CDC7 kinase, or any of them identified using liquid biopsy is also present in cancer cells present in a subject (e.g., in a tumor). In some embodiments, any of the types of dysregulation of expression, activity, or levels of the CDC7 gene, CDC7 kinase, or any of them described herein can be detected using liquid biopsies. In some embodiments, genetic mutations identified via liquid biopsy may be used to identify subjects as candidates for particular treatments.For example, detection of dysregulation of the expression or activity or levels of the CDC7 gene, CDC7 kinase, or any of them in a subject can indicate that the subject will respond to treatment comprising administration of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0174] Liquid biopsies can be performed multiple times during the course of diagnosis, monitoring, and / or treatment to determine one or more clinically relevant parameters, including, but not limited to, disease progression and / or treatment effectiveness. For example, a first liquid biopsy can be performed at a first time point, and a second liquid biopsy can be performed at a second time point during the course of diagnosis, monitoring, and / or treatment. In some embodiments, the first time point can be a time point before the subject is diagnosed with a disease (e.g., when the subject is healthy), and the second time point can be a time point after the subject has developed a disease (e.g., the second time point can be used to diagnose the subject with a disease). In some embodiments, the first time point can be a time point before the subject is diagnosed with a disease (e.g., when the subject is healthy), after which the subject is monitored, and the second time point can be a time point after the subject has been monitored. In some embodiments, the first time point can be a time point after the subject is diagnosed with a disease and then a treatment is administered to the subject, and the second time point can be a time point after the treatment is administered. In such cases, the second time point can be used to evaluate the effectiveness of the treatment (e.g., if the genetic mutation detected at the first time point is reduced in abundance or undetectable). In some embodiments, the treatment administered to the subject can include a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

[0175] In some embodiments, the efficacy of the compound of Formula (I) or a pharmaceutically acceptable salt thereof can be determined by assessing the allele frequency of dysregulated CDC7 genes in cfDNA obtained from the subject at different time points, for example, cfDNA obtained from the subject at a first time point and cfDNA obtained from the subject at a second time point, wherein at least one dose of the compound of Formula (I) or a pharmaceutically acceptable salt thereof is administered to the subject between the first and second time points. Some embodiments of these methods can further include administering at least one dose of the compound of Formula (I) or a pharmaceutically acceptable salt thereof to the subject between the first and second time points.For example, a reduction in the dysregulated allele frequency (AF) of the CDC7 gene in cfDNA obtained from the subject at a second time point compared to the dysregulated allele frequency (AF) of the CDC7 gene in cfDNA obtained from the subject at a first time point (e.g., 1% to about 99% reduction, 1% to about 95% reduction, 1% to about 90% reduction, 1% to about 85% reduction, 1% to about 80% reduction, 1% to about 75% reduction, 1% to about 70% reduction). , 1% reduction to approximately 65% ​​reduction, 1% reduction to approximately 60% reduction, 1% reduction to approximately 55% reduction, 1% reduction to approximately 50% reduction, 1% reduction to approximately 45% reduction, 1% reduction to approximately 40% reduction, 1% reduction to approximately 35% reduction, 1% reduction to approximately 30% reduction, 1% reduction to approximately 25% reduction, 1% reduction to approximately 20% reduction, 1% reduction to approximately 15% reduction, 1% reduction to approximately 10% reduction, 1% to approximately 5% reduction, approximately 5% to approximately 99% reduction, approximately 10% to approximately 99% reduction, approximately 1 5% to approximately 99% reduction, approximately 20% to approximately 99% reduction, approximately 25% to approximately 99% reduction, approximately 30% to approximately 99% reduction, approximately 35% to approximately 99% reduction, approximately 40% to approximately 99% reduction, approximately 45% to approximately 99% reduction, approximately 50% to approximately 99% reduction, approximately 55% to approximately 99% reduction, approximately 60% to approximately 99% reduction, approximately 65% ​​to approximately 99% reduction, approximately 70% to approximately 99% reduction, approximately 75% to approximately 95% reduction, approximately 80% to approximately 99% reduction, approximately 90% to approximately 99% reduction A decrease in AF (about 95% to about 99% reduction, about 5% to about 10% reduction, about 5% to about 25% reduction, about 10% to about 30% reduction, about 20% to about 40% reduction, about 25% to about 50% reduction, about 35% to about 55% reduction, about 40% to about 60% reduction, about 50% to about 75% reduction, about 60% to about 80% reduction, or about 65% to about 85% reduction) indicates that the compound of Formula (I) or a pharmaceutically acceptable salt thereof was effective in the subject. In some embodiments, the AF is reduced such that the level is below the detection limit of the instrument. Alternatively, an increase in the dysregulated allele frequency (AF) of the CDC7 gene in cfDNA obtained from the subject at a second time point compared to the dysregulated allele frequency (AF) of the CDC7 gene in cfDNA obtained from the subject at a first time point indicates that the compound of Formula (I) or a pharmaceutically acceptable salt thereof was not effective in the subject.Some embodiments of these methods may further include administering an additional dose of the compound of Formula (I) or a pharmaceutically acceptable salt thereof to a subject for whom the compound of Formula (I) or a pharmaceutically acceptable salt thereof has been determined to be effective. Some embodiments of these methods may further include administering a different treatment (e.g., a treatment that does not include administration of the compound of Formula (I) or a pharmaceutically acceptable salt thereof as monotherapy) to a subject for whom the compound of Formula (I) or a pharmaceutically acceptable salt thereof has been determined to be ineffective.

[0176] In some embodiments, the CDC7-associated cancer is a microsatellite instability-high (MSI-H) cancer. In other embodiments, the CDC7-associated cancer is not a microsatellite instability-high (MSI-H) cancer. In some embodiments, MSI-H status is determined by detection of repetitive DNA sequences selected from the group consisting of mononucleotide repeat markers, dinucleotide repeat markers, quasi-monomorphic markers, or a combination of any of the foregoing.

[0177] In some embodiments, the tumor associated with cancer comprises a phenotype selected from the group consisting of chromosomal instability (CIN), spindle checkpoint assembly defects, mitotic defects, Gl / S checkpoint defects, and combinations thereof. In some embodiments, the tumor associated with cancer comprises a Wnt signaling pathway mutation. In some embodiments, the Wnt signaling pathway mutation is selected from the group consisting of an adenomatous polyposis coli (APC) gene mutation, a FAT1 mutation, a FAT4 mutation, or a combination of any of the foregoing.

[0178] In some examples of these methods, the time difference between the first time point and the second time point is about 1 day to about 1 year, about 1 day to about 11 months, about 1 day to about 10 months, about 1 day to about 9 months, about 1 day to about 8 months, about 1 day to about 7 months, about 1 day to about 6 months, about 1 day to about 5 months, about 1 day to about 4 months, about 1 day to about 3 months, about 1 day to about 10 weeks, about 1 day to about 2 months, about 1 day to about 6 weeks, about 1 day to about 1 month, about 1 day to about 25 days, about 1 day to about 20 days, about 1 day to about 15 days, about 1 day to about 10 days, about 1 day to about 5 days, about 2 days to about 1 year, about 5 days to about 1 year, about 10 days to about 1 year, about 15 days to about 1 year, about 20 days to about 1 year, about 25 ... The period may be from days to about 1 year, from about 1 month to about 1 year, from about 6 weeks to about 1 year, from about 2 months to about 1 year, from about 3 months to about 1 year, from about 4 months to about 1 year, from about 5 months to about 1 year, from about 6 months to about 1 year, from about 7 months to about 1 year, from about 8 months to about 1 year, from about 9 months to about 1 year, from about 10 months to about 1 year, from about 11 months to about 1 year, from about 1 day to about 7 days, from about 1 day to about 14 days, from about 5 days to about 10 days, from about 5 days to about 20 days, from about 10 to about 20 days, from about 15 days to about 1 month, from about 15 days to about 2 months, from about 1 week to about 1 month, from about 2 weeks to about 1 month, from about 1 month to about 3 months, from about 3 months to about 6 months, from about 4 months to about 6 months, from about 5 months to about 8 months, or from about 7 months to about 9 months. In some embodiments of these methods, the subject may have previously been identified as having a cancer with a dysregulated CDC7 gene (e.g., any of the examples of dysregulated CDC7 genes described herein). In some embodiments of these methods, the subject may have previously been diagnosed with any of the types of cancer described herein. In some embodiments of these methods, the subject may have one or more metastases (e.g., one or more brain metastases).

[0179] In some of the above embodiments, the cfDNA comprises ctDNA, such as CDC7-associated ctDNA. For example, the cfDNA is ctDNA, such as CDC7-associated ctDNA. In some embodiments, at least some portion of the cfDNA is determined to be CDC7-associated ctDNA, e.g., a sequenced and / or quantified amount of total cfDNA is determined to have CDC7 fusion and / or CDC7 overexpression.

[0180] In the field of medical oncology, it is common practice to use a combination of different forms of treatment to treat each subject with cancer. In medical oncology, in addition to the compositions provided herein, other components of such conjoint treatments or therapies can be, for example, surgery, radiation therapy, and chemotherapeutic agents such as other kinase inhibitors, signal transduction inhibitors, and / or monoclonal antibodies. For example, surgery can be open surgery or minimally invasive surgery. Thus, the compounds of formula (I) or pharmaceutically acceptable salts thereof can also be useful as adjuvants for cancer treatment, i.e., they can be used in combination with one or more additional therapies or therapeutic agents, for example, chemotherapeutic agents that function by the same or different mechanisms of action. In some embodiments, the compounds of formula (I) or pharmaceutically acceptable salts thereof can be used before the administration of the additional therapeutic agent or therapy. For example, a subject in need thereof can be administered one or more doses of the compound of formula (I) or a pharmaceutically acceptable salt thereof for a certain period of time, followed by at least partial resection of the tumor. In some embodiments, treatment with one or more doses of a compound of Formula (I) or a pharmaceutically acceptable salt thereof reduces tumor size (e.g., tumor burden) prior to at least partial resection of the tumor. In some embodiments, a subject in need thereof may be administered one or more doses of a compound of Formula (I) or a pharmaceutically acceptable salt thereof over a period of time, followed by one or more rounds of radiation therapy. In some embodiments, treatment with one or more doses of a compound of Formula (I) or a pharmaceutically acceptable salt thereof reduces tumor size (e.g., tumor burden) prior to one or more rounds of radiation therapy.

[0181] In some embodiments, the subject has a cancer (e.g., a locally advanced or metastatic tumor) that is refractory or intolerant to standard therapy (e.g., administration of a chemotherapeutic agent such as a first CDC7 inhibitor or a multikinase inhibitor, immunotherapy, or radiation (e.g., radioactive iodine)). In some embodiments, the subject has a cancer (e.g., a locally advanced or metastatic tumor) that is refractory or intolerant to a previous therapy (e.g., administration of a chemotherapeutic agent such as a first CDC7 inhibitor or a multikinase inhibitor, immunotherapy, or radiation (e.g., radioactive iodine)). In some embodiments, the subject has a cancer (e.g., a locally advanced or metastatic tumor) that does not have a standard therapy. In some embodiments, the subject is CDC7 kinase inhibitor naive. For example, the subject is naive to treatment with a selective CDC7 kinase inhibitor. In some embodiments, the subject is not CDC7-kinase inhibitor naive.

[0182] In some embodiments, the subject has received prior therapy. In some embodiments, the subject with NSCLC (e.g., CDC7-associated NSCLS) has been treated with platinum-based chemotherapy, PD-1 / PDL1 immunotherapy, or both, before treatment with a compound of Formula (I), or a pharmaceutically acceptable salt thereof. In some embodiments, the subject with thyroid cancer (e.g., CDC7-associated thyroid cancer) has been treated with one or more of sorafenib, lenvatinib, and radioactive iodine before treatment with a compound of Formula (I), or a pharmaceutically acceptable salt thereof. In some embodiments, the subject with colorectal cancer (e.g., CDC7-associated colorectal cancer) has been treated with fluoropyrimidine-based chemotherapy, with or without anti-VEGF-directed therapy or anti-EGFR-directed therapy, before treatment with a compound of Formula (I), or a pharmaceutically acceptable salt thereof. In some embodiments, a subject with pancreatic cancer (e.g., CDC7-associated pancreatic cancer) has been treated with one or more of fluoropyrimidine-based chemotherapy, gemcitabine-based chemotherapy, and S-1 chemotherapy prior to treatment with a compound of Formula (I), or a pharmaceutically acceptable salt thereof. In some embodiments, a subject with breast cancer (e.g., CDC7-associated breast cancer) has been treated with one or more of anthracyclines, taxanes, HER2-directed therapy, and hormonal therapy prior to treatment with a compound of Formula (I), or a pharmaceutically acceptable salt thereof. In some embodiments, a subject with MTC (e.g., CDC7-associated MTC cancer) has been treated with one or more of caboxantinib and vandetanib prior to treatment with a compound of Formula (I), or a pharmaceutically acceptable salt thereof.

[0183] In some embodiments of any of the methods described herein, the compound of Formula (I), or a pharmaceutically acceptable salt thereof, is administered in combination with an effective amount of at least one additional therapeutic agent selected from one or more additional therapies or therapeutic agents (e.g., a chemotherapeutic agent).

[0184] Non-limiting examples of additional therapeutic agents include other CDC7-targeted therapeutic agents (i.e., first or second CDC7 kinase inhibitors), other kinase inhibitors (e.g., receptor tyrosine kinase-targeted therapeutic agents (e.g., Trk inhibitors or EGFR inhibitors)), signal transduction pathway inhibitors, checkpoint inhibitors, modulators of apoptosis pathways (e.g., obataclax), cytotoxic chemotherapeutic agents, angiogenesis-targeted therapies, immune-targeted agents including immunotherapies, and radiation therapy.

[0185] In some embodiments, the other CDC7 targeted therapeutic is a multikinase inhibitor that exhibits CDC7 inhibitory activity. In some embodiments, the other CDC7 targeted therapeutic inhibitor is selective for CDC7 kinase. Exemplary CDC7 kinase inhibitors have an inhibitory activity (IC) against CDC7 kinase of less than about 1000 nM, less than about 500 nM, less than about 200 nM, less than about 100 nM, less than about 50 nM, less than about 25 nM, less than about 10 nM, or less than about 1 nM, as measured in the assays described herein. 50 In some embodiments, the CDC7 kinase inhibitor can exhibit an inhibitory activity (IC) against CDC7 kinase of less than about 25 nM, less than about 10 nM, less than about 5 nM, or less than about 1 nM as measured in the assays provided herein. 50 ) can be shown.

[0186] Non-limiting examples of kinase-targeted therapeutic agents (e.g., first CDC7 inhibitors or second CDC7 inhibitors) include TAK931, SRA141, and PHA-767491.

[0187] Non-limiting examples of multikinase inhibitors include alectinib (9-ethyl-6,6-dimethyl-8-[4-(morpholin-4-yl)piperidin-1-yl]-11-oxo-6,11-dihydro-5H-benzo[b]carbazole-3-carbonitrile); amuvatinib (MP470, HPK56) (N-(1,3-benzodioxol-5-ylmethyl)-4-([1]benzofuro[3,2-d]pyrimidin-4-yl)piperazine-1-carbothioamide); apatinib (YN968D1) (N-[4-(1-cyanocyclopentyl)phenyl- 2-(4-picolyl)amino-3-nicotinamide methanesulfonate; cabozantinib (Cometrik XL-184) (N-(4-((6,7-dimethoxyquinolin-4-yl)oxy)phenyl)-N'-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide); dovitinib (TKI258; GFKI-258; CHIR-258) ((3Z)-4-amino-5-fluoro-3-[5-(4-methylpiperazin-1-yl)-1,3-dihydrobenzimidazol-2-ylidene]quinolin-2-one); famitinib (5- [2-(Diethylamino)ethyl]-2-[(Z)-(5-fluoro-2-oxo-1H-indol-3-ylidene)methyl]-3-methyl-6,7-dihydro-1H-pyrrolo[3,2-c]pyridin-4-one; fedratinib (SAR302503, TG101348) (N-(2-methyl-2-propanyl)-3-{[5-methyl-2-({4-[2-(1-pyrrolidinyl)ethoxy]phenyl}amino)-4-pyrimidinyl]amino}benzenesulfonamide); foCDC7inib (XL880, EXEL-2880, GSK136 3089, GSK089) (N1'-[3-fluoro-4-[[6-methoxy-7-(3-morpholinopropoxy)-4-quinolyl]oxy]phenyl]-N1-(4-fluorophenyl)cyclopropane-1,1-dicarboxamide); fostamantinib (R788) (2H-pyrido[3,2-b]-1,4-oxazin-3(4H)-one, 6-[[5-fluoro-2-[(3,4,5-trimethoxyphenyl)amino]-4-pyrimidinyl]amino]-2,2-dimethyl-4-[(phosphonooxy)methyl]-, sodium salt (1:2));Ilorasertib (ABT-348) (1-(4-(4-amino-7-(1-(2-hydroxyethyl)-1H-pyrazol-4-yl)thieno[3,2-c]pyridin-3-yl)phenyl)-3-(3-fluorophenyl)urea); lenvatinib (E7080, Lenvima) (4-[3-chloro-4-(cyclopropylaminocarbonyl)aminophenoxy]-7-methoxy-6-quinolinecarboxamide); motesanib (AMG706) (N-(3,3-dimethyl-2,3-dihydro-1H-indol-6-yl)-2-[( pyridin-4-ylmethyl)amino]pyridine-3-carboxamide; nintedanib (3-Z-[1-(4-(N-((4-methyl-piperazin-1-yl)-methylcarbonyl)-N-methyl-amino)-anilino)-1-phenyl-methylene]-6-methoxycarbonyl-2-indolinone); ponatinib (AP24534) (3-(2-imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-[4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl]benzamide) PP242 (Tolquinib) (2-[4-amino-1-(1-methylethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]-1H-indol-5-ol); Quizartinib (1-(5-(tert-butyl)isoxazol-3-yl)-3-(4-(7-(2-morpholinoethoxy)benzo[d]imidazo[2,1-b]thiazol-2-yl)phenyl)urea); Regorafenib (BAY73-4506, Stivarga) (4-[4-({[4-chloro-3-(trifluoromethyl)phenyl]carbamoyl]carbamoyl) yl}amino)-3-fluorophenoxy]-N-methylpyridine-2-carboxamide hydrate; RXDX-105 (CEP-32496, agerafenib) (1-(3-((6,7-dimethoxyquinazolin-4-yl)oxy)phenyl)-3-(5-(1,1,1-trifluoro-2-methylpropan-2-yl)isoxazol-3-yl)urea); Semaxanib (SU5416) ((3Z)-3-[(3,5-dimethyl-1H-pyrrol-2-yl)methylidene]-1,3-dihydro-2H-indol-2-one);Sitravatinib (MGCD516, MG516) (N-(3-fluoro-4-{[2-(5-{[(2-methoxyethyl)amino]methyl}-2-pyridinyl)thieno[3,2-b]pyridin-7-yl]oxy}phenyl)-N'-(4-fluorophenyl)-1,1-cyclopropanedicarboxamide); sorafenib (BAY43-9006) (4-[4-[[[[4-chloro-3-(trifluoromethyl)phenyl]amino]carbonyl]amino]phenoxy]-N-methyl-2-pyridinecarboxamide); vandetanib (N-(4-bromo 1-(4-(4-amino-1-propane-2-yl)phenyl)-6-methoxy-7-[(1-methylpiperidin-4-yl)methoxy]quinazolin-4-amine; Vatalanib (PTK787, PTK / ZK, ZK222584) (N-(4-chlorophenyl)-4-(pyridin-4-ylmethyl)phthalazin-1-amine); AD-57 (N-[4-[4-amino-1-(1-methylethyl)-1H-pyrazolo[3,4-d]pyrimidin-3-yl]phenyl]-N'-[3-(trifluoromethyl)phenyl]-urea); AD-80 (1-[4-(4-amino-1-propane-2-yl)phenyl]- 1-(4-(4-amino-1-isopropyl-1H-pyrazolo[3,4-d]pyrimidin-3-yl)phenyl)-3-(4-chloro-3-(trifluoromethyl)phenyl)urea; ALW-II-41-27 (N-(5-((4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)carbamoyl)-2-methylphenyl)-5-(thiophen-2-yl) Nicotinamide; BPR1K871 (1-(3-chlorophenyl)-3-(5-(2-((7-(3-(dimethylamino)propoxy)quinazolin-4-yl)amino)ethyl)thiazol-2-yl)urea); CLM3 (1-phenethyl-N-(1-phenylethyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine); EBI-907 (N-(2-chloro-3-(1-cyclopropyl-8-methoxy-3H-pyrazolo[3,4-c]isoquinolin-7-yl)-4-fluorophenyl)-3-fluoropropane-1-sulfonamide);NVP-AST-487 (N-[4-[(4-ethyl-1-piperazinyl)methyl]-3-(trifluoromethyl)phenyl]-N'-[4-[[6-(methylamino)-4-pyrimidinyl]oxy]phenyl]-urea); NVP-BBT594 (BBT594) (5-((6-acetamidopyrimidin-4-yl)oxy)-N-(4-((4-methylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)indoline-1-carboxamide); PD173955 (6-(2,6-dichlorophenyl)-8 -methyl-2-(3-methylsulfanylanilino)pyrido[2,3-d]pyrimidin-7-one; PP2 (4-amino-5-(4-chlorophenyl)-7-(dimethylethyl)pyrazolo[3,4-d]pyrimidine); PZ-1 (N-(5-(tert-butyl)isoxazol-3-yl)-2-(4-(5-(1-methyl-1H-pyrazol-4-yl)-1H-benzo[d]imidazol-1-yl)phenyl)acetamide); RPI-1 (1,3-dihydro-5,6-dimethoxy-3-[(4-hydroxyphenyl) methylene]-H-indol-2-one; (3E)-3-[(4-hydroxyphenyl)methylidene]-5,6-dimethoxy-1H-indol-2-one; SGI-7079 (3-[2-[[3-fluoro-4-(4-methyl-1-piperazinyl)phenyl]amino]-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl]-benzeneacetonitrile); SPP86 (1-isopropyl-3-(phenylethynyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine); SU4984 (4-[4-[(E) -(2-oxo-1H-indol-3-ylidene)methyl]phenyl]piperazine-1-carbaldehyde; sunitinb (SU11248) (N-(2-diethylaminoethyl)-5-[(Z)-(5-fluoro-2-oxo-1H-indol-3-ylidene)methyl]-2,4-dimethyl-1H-pyrrole-3-carboxamide); TG101209 (N-tert-butyl-3-(5-methyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)pyrimidin-4-ylamino)benzenesulfonamide);Withaferin A ((4β,5β,6β,22R)-4,27-dihydroxy-5,6:22,26-diepoxyergosta-2,24-diene-1,26-dione); XL-999 ((Z)-5-((1-ethylpiperidin-4-yl)amino)-3-((3-fluorophenyl)(5-methyl-1H-imidazol-2-yl)methylene)indolin-2-one); BPR1J373 (5-phenylthiazol-2-ylamine-pyriminide derivative); CG-806 (CG'806); DCC-2157; G TX-186; HG-6-63-01 ((E)-3-(2-(4-chloro-1H-pyrrolo[2,3-b]pyridin-5-yl)vinyl)-N-(4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)-4-methylbenzamide); SW-01 (cyclobenzaprine hydrochloride); XMD15-44 (N-(4-((4-ethylpiperazin-1-yl)methyl)-3-(trifluoromethyl)phenyl)-4-methyl-3-(pyridin-3-ylethynyl)benzamide (structure) Y078-DM1 (an antibody-drug conjugate composed of a CDC7 antibody (Y078) conjugated to a derivative of the cytotoxic drug maytansine); Y078-DM4 (an antibody-drug conjugate composed of a CDC7 antibody (Y078) conjugated to a derivative of the cytotoxic drug maytansine); ITRI-305 (D0N5TB, DIB003599); BLU-667 ((1S,4R)-N-((S)-1-(6-(4-fluoro-1H-pyrazol-1-yl)pyridin-3-yl)ethyl)-1-methoxy-4-(4-methyl- 6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)cyclohexane-1-carboxamide; BLU6864; DS-5010; GSK3179106; GSK3352589; NMS-E668; TAS0286 / HM05; TPX0046; and N-(3-(2-(dimethylamino)ethoxy)-5-(trifluoromethyl)phenyl)-2-(4-(4-ethoxy-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorophenyl)acetamide;

[0188] Non-limiting examples of receptor tyrosine kinase (e.g., Trk) targeted therapeutic agents include afatinib, cabozantinib, cetuximab, crizotinib, dabrafenib, entrectinib, erlotinib, gefitinib, imatinib, lapatinib, lestaurtinib, nilotinib, pazopanib, panitumumab, pertuzumab, sunitinib, trastuzumab, 1-((3S,4R)-4-(3-fluorophenyl)-1-(2-methoxyethyl)pyrrolidin-3-yl)-3-(4-methyl-3-(2-methylpyrimidin-5-yl)-1-phenyl-1H-pyrazol-5-yl)urea, AG 879, AR-772, AR-786, AR-256, AR-618, AZ-23, AZ623, DS-6051, Go 6976, GNF-5837, GTx-186, GW 441756, LOXO-101, MGCD516, PLX7486, RXDX101, VM-902A, TPX-0005, TSR-011, GNF-4256, N-[3-[[2,3-dihydro-2-oxo-3-(1H-pyrrol-2-ylmethylene)-1H-indol-6-yl]amino]-4-methylphenyl]-N'-[2-fluoro-5-(trifluoromethyl)phenyl]-urea, AZ623, AZ64, (S)-5-chloro-N2- (1-(5-fluoropyridin-2-yl)ethyl)-N4-(5-isopropoxy-1H-pyrazol-3-yl)pyrimidine-2,4-diamine, AZD7451, CEP-751, CT327, sunitinib, GNF-8625, and (R)-1-(6-(6-(2-(3-fluorophenyl)pyrrolidin-1-yl)imidazo[1,2-b]pyridazin-3-yl)-[2,4'-bipyridin]-2'-yl)piperidin-4-ol.

[0189] Non-limiting examples of BRAF inhibitors include dabrafenib, vemurafenib (also known as RG7204 or PLX4032), sorafenib tosylate, PLX-4720, GDC-0879, BMS-908662 (Bristol-Meyers Squibb), LGX818 (Novartis), PLX3603 (Hofmann-LaRoche), RAF265 (Novartis), RO5185426 (Hofmann-LaRoche), and GSK2118436 (GlaxoSmithKline). Additional examples of BRAF inhibitors are known in the art.

[0190] In some embodiments, the receptor tyrosine kinase inhibitor is an epidermal growth factor receptor tyrosine kinase inhibitor (EGFR). For example, EGFR inhibitors can include osimertinib (merelectinib, Tagrisso), erlotinib (Tarceva), gefitinib (Iressa), cetuximab (Erbitux), necitumumab (Portrazza), neratinib (Nerlynx), lapatinib (Tykerb), panitumumab (Vectibix), and vandetanib (Caprelsa).

[0191] In some embodiments, signal transduction pathway inhibitors include Ras-Raf-MEK-ERK pathway inhibitors (e.g., binimetinib, selumetinib, encorafenib, sorafenib, trametinib, and vemurafenib), PI3K-Akt-mTOR-S6K pathway inhibitors (e.g., everolimus, rapamycin, perifosine, temsirolimus), and baricitinib, Brigatinib, capmatinib, danusertib, ibrutinib, mirciclib, quercetin, regorafenib, ruxolitinib, semaxanib, AP32788, BLU285, BLU554, INCB39110, INCB40093, INCB50465, INCB52793, INCB54828, MGCD265, NMS-088, NMS-1286937, PF Other kinase inhibitors include 477736 ((R)-amino-N-[5,6-dihydro-2-(1-methyl-1H-pyrazol-4-yl)-6-oxo-1H-pyrrolo[4,3,2-ef][2,3]benzodiazepin-8-yl]-cyclohexaneacetamide), PLX3397, PLX7486, PLX8394, PLX9486, PRN1008, PRN1371, RXDX103, RXDX106, RXDX108, and TG101209 (N-tert-butyl-3-(5-methyl-2-(4-(4-methylpiperazin-1-yl)phenylamino)pyrimidin-4-ylamino)benzenesulfonamide).

[0192] Non-limiting examples of checkpoint inhibitors include ipilimumab, tremelimumab, nivolumab, pidilizumab, MPDL3208A, MEDI4736, MSB0010718C, BMS-936559, BMS-956559, BMS-935559 (MDX-1105), AMP-224, and pembrolizumab.

[0193] In some embodiments, the cytotoxic chemotherapeutic agent is selected from arsenic trioxide, bleomycin, cabazitaxel, capecitabine, carboplatin, cisplatin, cyclophosphamide, cytarabine, dacarbazine, daunorubicin, docetaxel, doxorubicin, etoposide, fluorouracil, gemcitabine, irinotecan, lomustine, methotrexate, mitomycin C, oxaliplatin, paclitaxel, pemetrexed, temozolomide, and vincristine.

[0194] Non-limiting examples of angiogenesis targeted therapeutic agents include aflibercept and bevacizumab.

[0195] In some embodiments, the additional therapeutic or therapeutic agent can comprise a histidyl-tRNA synthetase (HRS) polypeptide or an expressible nucleotide encoding an HRS polypeptide.

[0196] The term "immunotherapy" refers to an agent that modulates the immune system. In some embodiments, immunotherapy can increase the expression and / or activity of regulators of the immune system. In some embodiments, immunotherapy can decrease the expression and / or activity of regulators of the immune system. In some embodiments, immunotherapy can recruit and / or enhance the activity of immune cells.

[0197] In some embodiments, the immunotherapy is a cellular immunotherapy (e.g., adoptive T cell therapy, dendritic cell therapy, natural killer cell therapy). In some embodiments, the cellular immunotherapy is sipuleucel-T (APC8015, Provenge™, see Plosker (2011) Drugs 71(1):101-108). In some embodiments, the cellular immunotherapy comprises cells expressing a chimeric antigen receptor (CAR). In some embodiments, the cellular immunotherapy is a CAR-T cell therapy. In some embodiments, the CAR-T cell therapy is tisagenlecleucel (Kymriah™).

[0198] In some embodiments, the immunotherapy is an antibody therapy (e.g., monoclonal antibody, conjugated antibody). In some embodiments, the antibody therapy is selected from the group consisting of bevacizumab (Mvasti™, Avastin®), trastuzumab (Herceptin®), avelumab (Bavencio®), rituximab (MabThera™, Rituxan®), edrecolomab (Panorex), daratumab (Darzalex®), olaratumab (Lartruvo™), ofatumumab (Arzerra®), alemtuzumab (Campath®), cetuximab (Erbitux®), oregovomab, pembrolizumab (Keytruda®), dinutiximab (Unituxin®), obinutuzumab (Gazyva®), and the like. )), tremelimumab (CP-675,206), ramucirumab (Cyramza®), ublituximab (TG-1101), panitumumab (Vectibix®), elotuzumab (Empliciti™), avelumab (Bavencio®), necitumumab (Portrazza™), cirumutuzumab (UC-961), ibritumomab (Zevalin®), isatuximab (SAR650984), nimotuzumab, fresolimumab (GC1008), lirilumab (INN), mogamulizumab (Poteligeo®), ficlatuzumab (AV-299), denosumab (Xgeva®), ganitumab, urelumab, pidilizumab, or amatuximab.

[0199] In some embodiments, the immunotherapy is an antibody-drug conjugate. In some embodiments, the antibody-drug conjugate is gemtuzumab ozogamicin (Mylotarg™), inotuzumab ozogamicin (Besponsa®), brentuximab vedotin (Adcetris®), ado-trastuzumab emtansine (TDM-1, Kadcyla®), mirvetuximab soravtansine (IMGN853), or anetumab ravtansine.

[0200] In some embodiments, the immunotherapy comprises blinatumomab (AMG103, Blincyto®) or midostaurin (Rydapt).

[0201] In some embodiments, the immunotherapy comprises a toxin, hi some embodiments, the immunotherapy is denileukin diftitox (Ontak®).

[0202] In some embodiments, the immunotherapy is cytokine therapy. In some embodiments, the cytokine therapy is interleukin-2 (IL-2) therapy, interferon alpha (IFNα) therapy, granulocyte colony-stimulating factor (G-CSF) therapy, interleukin-12 (IL-12) therapy, interleukin-15 (IL-15) therapy, interleukin-7 (IL-7) therapy, or erythropoietin alpha (EPO) therapy. In some embodiments, the IL-2 therapy is aldesleukin (Proleukin®). In some embodiments, the IFNα therapy is IntronA® (Roferon-A®). In some embodiments, the G-CSF therapy is filgrastim (Neupogen®).

[0203] In some embodiments, the immunotherapy is an immune checkpoint inhibitor. In some embodiments, the immunotherapy comprises one or more immune checkpoint inhibitors. In some embodiments, the immune checkpoint inhibitor is a CTLA-4 inhibitor, a PD-1 inhibitor, or a PD-L1 inhibitor. In some embodiments, the CTLA-4 inhibitor is ipilimumab (Yervoy®) or tremelimumab (CP-675,206). In some embodiments, the PD-1 inhibitor is pembrolizumab (Keytruda®) or nivolumab (Opdivo®). In some embodiments, the PD-L1 inhibitor is atezolizumab (Tecentriq®), avelumab (Bavencio®), or durvalumab (Imfinzi™).

[0204] In some embodiments, the immunotherapy is an mRNA-based immunotherapy. In some embodiments, the mRNA-based immunotherapy is CV9104 (see, e.g., Rausch et al. (2014) Human Vaccin Immunother 10(11):3146-52, and Kubler et al. (2015) J. Immunother Cancer 3:26).

[0205] In some embodiments, the immunotherapy is bacillus Calmette-Guerin (BCG) therapy.

[0206] In some embodiments, the immunotherapy is oncolytic virus therapy. In some embodiments, the oncolytic virus therapy is talimogene alherparepvec (T-VEC, Imlygic®).

[0207] In some embodiments, the immunotherapy is a cancer vaccine. In some embodiments, the cancer vaccine is a human papillomavirus (HPV) vaccine. In some embodiments, the HPV vaccine is Gardasil®, Gardasil 9®, or Cervarix®. In some embodiments, the cancer vaccine is a hepatitis B virus (HBV) vaccine. In some embodiments, the HBV vaccine is Engerix-B®, Recombivax HB®, or GI-13020 (Tarmogen®). In some embodiments, the cancer vaccine is Twinrix® or Pediarix®. In some embodiments, the cancer vaccine is BiovaxID®, Oncophage®, GVAX, ADXS11-001, ALVAC-CEA, PROSTVAC®, Rindopepimut®, CimaVax-EGF, lapleucel-T (APC8024, Neuvenge™), GRNVAC1, GRNVAC2, GRN-1201, hepcortespenlisimut-L (Hepko-V5), DCVAX®, SCIB1, BMT CTN 1401, PrCa VBIR, PANVAC, ProstAtak®, DPX-Survivac, or viagenpumatucel-L (HS-110).

[0208] In some embodiments, the immunotherapy is a peptide vaccine. In some embodiments, the peptide vaccine is nelipepimut-S(E75) (NeuVax™), IMA901, or SurVaxM (SVN53-67). In some embodiments, the cancer vaccine is an immunogenic personalized neoantigen vaccine (see, e.g., Ott et al. (2017) Nature 547:217-221; Sahin et al. (2017) Nature 547:222-226). In some embodiments, the cancer vaccine is RGSH4K or NEO-PV-01. In some embodiments, the cancer vaccine is a DNA-based vaccine. In some embodiments, the DNA-based vaccine is a mammaglobin-A DNA vaccine (see, e.g., Kim et al. (2016) OncoImmunology 5(2):e1069940).

[0209] In some embodiments, the immune-targeted agent is selected from aldesleukin, interferon alpha-2b, ipilimumab, lambrolizumab, nivolumab, prednisone, and sipuleucel-T.

[0210] Non-limiting examples of radiation therapy include radioactive iodide therapy, external beam radiation, and radium-223 therapy.

[0211] Additional kinase inhibitors are described, for example, in U.S. Patent Nos. 7,514,446, 7,863,289, 8,026,247, 8,501,756, 8,552,002, 8,815,901, 8,912,204, 9,260,437, 9,273,051, U.S. Publication No. 2015 / 0018336, International Publication Nos. 2007 / 002325, 2007 / 002433, 2008 / 080001, 2008 / 079906, 2008 / 079903, 2008 / 079909, 2008 / 080015, 2009 / 007748, 2009 / 012283, 2009 / 143018, 2009 / 143024, 2009 / 014637, 2009 / 152083, 2010 / 111527, 2012 / 109075, 2014 / 194127, 2015 / 112806, 2007 / 110344, 2009 / 071480, 2009 / 118411, 201 0 / 031816, 2010 / 145998, 2011 / 092120, 2012 / 101032, 2012 / 139930, 2012 / 143248, 2012 / 152763, 2013 / 014 No. 039, No. 2013 / 102059, No. 2013 / 050448, No. 2013 / 050446, No. 2014 / 019908, No. 2014 / 072220, No. 2014 / 184069, No. 2016 / 075224, Same No. 2016 / 081450, No. 2016 / 022569, No. 2016 / 011141, No. 2016 / 011144, No. 2016 / 011147, No. 2015 / 191667, No. 2012 / 101029, No. 201 2 / 113774, 2015 / 191666, 2015 / 161277, 2015 / 161274, 2015 / 108992, 2015 / 061572, 2015 / 058129, 2015 / 057873, 2015 / 017528, 2015 / 017533, 2014 / 160521, and 2014 / 011900, each of which is incorporated herein by reference in its entirety.

[0212] Although the genetic basis of tumorigenesis may vary among different cancer types, the cellular and molecular mechanisms required for metastasis appear to be similar across all solid tumor types. During the metastatic cascade, cancer cells lose growth inhibitory responses, undergo changes in adhesion, and produce enzymes that can degrade extracellular matrix components. This leads to tumor cell detachment from the original tumor, invasion into the circulation through newly formed vasculature, and migration and extravasation at favorable distant sites where they can colonize. Several genes have been identified as promoters or suppressors of metastasis. For example, overexpression of glial cell line-derived neurotrophic factor (GDNF) and its CDC7 receptor tyrosine kinase has been correlated with cancer growth and metastasis. See, e.g., Zeng, et al. J. Int. Med. Res. (2008) 36(4):656-64.

[0213] Therefore, also provided herein is a method for inhibiting, preventing, or assisting in the prevention of cancer metastasis or reducing symptoms in a subject in need thereof, comprising administering to the subject an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. Such a method can be used to treat one or more of the cancers described herein. For example, see U.S. Publication No. 2013 / 0029925, WO 2014 / 083567, and U.S. Patent No. 8,568,998. See, for example, Hezam K et al., Rev Neurosci 2018 Jan 26;29:93-98; Gao L, et al., Pancreas 2015 Jan;44:134-143; Ding K et al., J Biol Chem 2014 Jun 6;289:16057-71; and Amit M et al., Oncogene 2017 Jun 8;36:3232-3239. In some embodiments, the cancer is a CDC7-associated cancer. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, is used in combination with another therapeutic agent, including an additional therapy or chemotherapeutic agent, such as a kinase inhibitor. For example, a first or second CDC7 kinase inhibitor. In some embodiments, the additional therapeutic agent is crizotinib. In some embodiments, the additional therapeutic agent is osimertinib. In some embodiments, the subject has received one or more doses of a compound of Formula (I) or a pharmaceutically acceptable salt thereof prior to administration of the pharmaceutical composition. In some embodiments, the cancer is lung cancer (e.g., CDC7-associated lung cancer). In some embodiments, the additional therapeutic agent is a PARP inhibitor (e.g., olaparib). In some embodiments, the additional therapeutic agent is an ATR inhibitor (e.g., selalasertib). In some embodiments, the additional therapeutic agent is a Wee1 inhibitor (e.g., AZD-1775). In some embodiments, the additional therapeutic agent is an EGFR inhibitor (e.g., lapatinib).

[0214] The term "metastasis" is an art-recognized term that refers to the formation of additional tumors (e.g., solid tumors) in a subject at a site distant from the primary tumor, wherein the additional tumors contain the same or similar cancer cells as the primary tumor.

[0215] Also provided are methods for reducing the risk of developing metastasis or additional metastases in a subject with a CDC7-associated cancer, comprising selecting, identifying, or diagnosing the subject as having a CDC7-associated cancer and administering an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof to the subject selected, identified, or diagnosed as having a CDC7-associated cancer. Also provided are methods for reducing the risk of developing metastasis or additional metastases in a subject with a CDC7-associated cancer, comprising administering an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof to the subject. The reduced risk of developing metastasis or additional metastases in a subject with a CDC7-associated cancer can be compared to the risk of developing metastasis or additional metastases in the subject before treatment, or compared to a subject or population of subjects with a similar or identical CDC7-associated cancer who have not received treatment or who have received a different treatment. In some embodiments, the additional therapeutic agent is crizotinib. In some embodiments, the additional therapeutic agent is osimertinib. In some embodiments, the subject has received one or more doses of a compound of Formula (I) or a pharmaceutically acceptable salt thereof prior to administration of the pharmaceutical composition. In some embodiments, the cancer is lung cancer (e.g., CDC7-associated lung cancer).

[0216] The phrase "risk of developing metastases" refers to the risk that a subject with a primary tumor will develop additional tumors (e.g., solid tumors) at sites distant from the primary tumor in the subject over a period of time, where the additional tumors contain the same or similar cancer cells as the primary tumor. Methods for reducing the risk of developing metastases in a subject with cancer are described herein.

[0217] The phrase "risk of developing additional metastases" refers to the risk that a subject who has a primary tumor and one or more additional tumors at sites distant from the primary tumor (where the one or more additional tumors contain the same or similar cancer cells as the primary tumor) will develop one or more additional tumors distant from the primary tumor, where the additional tumors contain the same or similar cancer cells as the primary tumor. Methods for reducing the risk of developing additional metastases are described herein.

[0218] Treatment of a subject with cancer with a multikinase inhibitor (MKI) or a target-specific kinase inhibitor (e.g., a BRAF inhibitor, an EGFR inhibitor, a MEK inhibitor, an ALK inhibitor, a ROS1 inhibitor, a MET inhibitor, an aromatase inhibitor, a RAF inhibitor, a RET inhibitor, or a RAS inhibitor) can result in dysregulation of the CDC7 gene, CDC7 kinase, or their expression, activity, or levels in the cancer, and / or resistance to the CDC7 inhibitor. See, for example, Bhinge et al., Oncotarget 8:27155-27165, 2017; Chang et al., Yonsei Med. J. 58:9-18, 2017; and Lopez-Delisle et al., doi:10.1038 / s41388-017-0039-5, Oncogene 2018.

[0219] Treatment of a subject having cancer with a CDC7 inhibitor in combination with a multikinase inhibitor or a target-specific kinase inhibitor (e.g., a BRAF inhibitor, an EGFR inhibitor, a MEK inhibitor, an ALK inhibitor, a ROS1 inhibitor, a MET inhibitor, an aromatase inhibitor, a RAF inhibitor, a RET inhibitor, or a RAS inhibitor) may result in an enhanced therapeutic effect compared to treatment of the same or a similar subject with the CDC7 inhibitor as monotherapy, or with the multikinase inhibitor or the target-specific kinase inhibitor as monotherapy. See, e.g., Tang et al., doi:10.1038 / modpathol.2017.109, Mod. Pathol. 2017; Andreucci et al., Oncotarget 7:80543-80553, 2017; Nelson-Taylor et al., Mol. Cancer Ther. 16:1623-1633, 2017; and Kato et al., Clin. Cancer Res. 23:1988-1997, 2017.

[0220] Provided herein are methods of treating a subject having cancer (e.g., any of the cancers described herein) and who has previously been administered a multikinase inhibitor (MKI) or a target-specific kinase inhibitor (e.g., a BRAF inhibitor, an EGFR inhibitor, a MEK inhibitor, an ALK inhibitor, a ROS1 inhibitor, a MET inhibitor, an aromatase inhibitor, a RAF inhibitor, a RET inhibitor, or a RAS inhibitor) (e.g., as monotherapy), the methods comprising administering to the subject (i) an effective dose of a compound of Formula (I) or a pharmaceutically acceptable salt thereof as monotherapy, or (ii) an effective dose of a compound of Formula (I) or a pharmaceutically acceptable salt thereof and effective doses of the previously administered MKI or the previously administered target-specific kinase inhibitor.

[0221] Provided herein are methods of treating a subject having cancer (e.g., any of the cancers described herein) who has previously been administered an MKI or target-specific kinase inhibitor (e.g., a BRAF inhibitor, an EGFR inhibitor, a MEK inhibitor, an ALK inhibitor, a ROS1 inhibitor, a MET inhibitor, an aromatase inhibitor, a RAF inhibitor, a RET inhibitor, or a RAS inhibitor) (e.g., as monotherapy), the method comprising: identifying a subject having cancer cells with dysregulation of the CDC7 gene, CDC7 kinase, or their expression or activity or levels; and administering to the identified subject (i) an effective dose of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, as monotherapy, or (ii) an effective dose of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and effective doses of the previously administered MKI or the previously administered target-specific kinase inhibitor.

[0222] Provided herein are methods of treating a subject having cancer (e.g., any of the cancers described herein), the method comprising administering an effective amount of an MKI or target-specific kinase inhibitor (e.g., a BRAF inhibitor, an EGFR inhibitor, a MEK inhibitor, an ALK inhibitor, a ROS1 inhibitor, a MET inhibitor, an aromatase inhibitor, a RAF inhibitor, a RET inhibitor, or a RAS inhibitor) (e.g., as monotherapy) to the subject for a first period of time, and after that period of time, identifying a subject having cancer cells with dysregulated CDC7 gene, CDC7 kinase, or their expression, activity, or levels, and administering to the identified subject (i) an effective dose of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, or (ii) an effective dose of a compound of Formula (I) or a pharmaceutically acceptable salt thereof, and effective doses of a previously administered MKI or a previously administered target-specific kinase inhibitor.

[0223] Also provided is a method for inhibiting CDC7 kinase activity in a mammalian cell, the method comprising contacting the mammalian cell with a compound of Formula (I). In some embodiments, the contacting is in vitro. In some embodiments, the contacting is in vivo. In some embodiments, the contacting is in vivo and the method comprises administering an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof to a subject having a mammalian cell with CDC7 kinase activity. In some embodiments, the mammalian cell is a mammalian cancer cell. In some embodiments, the mammalian cancer cell is any cancer described herein. In some embodiments, the mammalian cancer cell is a CDC7-associated mammalian cancer cell.

[0224] Also provided is a method for inhibiting CDC7 kinase activity in a mammalian cell, the method comprising contacting the mammalian cell with a compound of Formula (I). In some embodiments, the contacting is in vitro. In some embodiments, the contacting is in vivo. In some embodiments, the contacting is in vivo and the method comprises administering an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof to a mammal having a mammalian cell with CDC7 kinase activity. In some embodiments, the mammalian cell is a mammalian cancer cell. In some embodiments, the mammalian cancer cell is any cancer described herein. In some embodiments, the mammalian cancer cell is a CDC7-associated mammalian cancer cell. In some embodiments, the mammalian cell is a gastrointestinal mammalian cell.

[0225] As used herein, the term "contacting" refers to bringing together the indicated moieties in an in vitro system or an in vivo system. For example, "contacting" CDC7 kinase with a compound provided herein includes administering a compound provided herein to a subject, such as a human, having CDC7 kinase, and introducing a compound provided herein into a sample containing, for example, a mammalian cell or purified preparation containing CDC7 kinase.

[0226] Also provided herein is a method of inhibiting the proliferation of mammalian cells in vitro or in vivo, comprising contacting the mammalian cells with an effective amount of a compound of formula (I), as defined herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0227] As defined herein, a "CDC7 kinase inhibitor" includes any compound that exhibits CDC7 inhibitory activity. In some embodiments, the CDC7 kinase inhibitor is selective for CDC7 kinase. Exemplary CDC7 kinase inhibitors have an inhibitory activity (IC) against CDC7 kinase of less than about 1000 nM, less than about 500 nM, less than about 200 nM, less than about 100 nM, less than about 50 nM, less than about 25 nM, less than about 10 nM, or less than about 1 nM, as measured in the assays described herein. 50 In some embodiments, the CDC7 kinase inhibitor can exhibit an inhibitory activity (IC) against CDC7 kinase of less than about 25 nM, less than about 10 nM, less than about 5 nM, or less than about 1 nM as measured in the assays provided herein. 50 ) can be shown.

[0228] As used herein, a "first CDC7 kinase inhibitor" or "first CDC7 inhibitor" is a CDC7 kinase inhibitor as defined herein, but does not include a compound of Formula (I) as defined herein, or a pharmaceutically acceptable salt thereof. As used herein, a "second CDC7 kinase inhibitor" or "second CDC7 inhibitor" is a CDC7 kinase inhibitor as defined herein, but does not include a compound of Formula (I) as defined herein, or a pharmaceutically acceptable salt thereof. When both a first and a second CDC7 inhibitor are present in the methods provided herein, the first and second CDC7 kinase inhibitors are different.

[0229] Exemplary first and second CDC7 kinase inhibitors are described herein. In some embodiments, the first or second CDC7 kinase inhibitor can be selected from the group consisting of TAK931, SRA141, and PHA-767491.

[0230] The phrase "effective amount" refers to an amount sufficient, when administered to a subject in need of such treatment, to (i) treat a CDC7-associated disease or disorder (such as a CDC7-associated cancer), (ii) attenuate, ameliorate, or eliminate one or more symptoms of a particular disease, condition, or disorder, or (iii) delay the onset of one or more symptoms of a particular disease, condition, or disorder described herein. The amount of the compound of formula (I), or a pharmaceutically acceptable salt thereof that would correspond to such an amount, will vary depending on factors such as the particular compound, the disease state and its severity, and the individual characteristics (e.g., body weight) of the subject in need of treatment, but can nevertheless be routinely determined by one of ordinary skill in the art.

[0231] When used as a pharmaceutical, the compounds of formula (I), including their pharmaceutically acceptable salts, can be administered in the form of pharmaceutical compositions. These compositions can be prepared by methods well known in the pharmaceutical art and can be administered by various routes, depending on whether local or systemic treatment is desired and the area to be treated. Administration can be topical (including transdermal, epidermal, ophthalmic, and mucosal, including intranasal, vaginal, and rectal delivery), pulmonary (e.g., intratracheal or intranasal, by inhalation or insufflation of powders or aerosols, including by nebulizer), oral, or parenteral. Oral administration can include dosage forms formulated for once-daily or twice-daily (BID) administration. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular, or injection or infusion, or intracranial, e.g., intrathecal or intraventricular, administration. Parenteral administration can be in the form of a single bolus dose or, for example, by a continuous perfusion pump. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners, and the like may be necessary or desirable.

[0232] The present invention also provides pharmaceutical compositions containing, as an active ingredient, a compound of Formula (I) or a pharmaceutically acceptable salt thereof, in combination with one or more pharmaceutically acceptable excipients. In some embodiments, the compound of Formula (I) or a pharmaceutically acceptable salt thereof present in the composition is a compound of Formula (IA), (IB), (IC), (ID), (IE), (IF), (IG), (IH), (II), (IJ), (IK), (IL), (IM), (IN), (IO), or (IP), or a pharmaceutically acceptable salt of any of the foregoing.

[0233] For example, a pharmaceutical composition prepared using a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the composition is suitable for topical administration. In preparing the compositions provided herein, the active ingredient is typically mixed with an excipient, diluted by an excipient, or enclosed within such a carrier, for example, in the form of a capsule, sachet, paper, or other container. When the excipient serves as a diluent, it can be a solid, semi-solid, or liquid substance that acts as a vehicle, carrier, or medium for the active ingredient. Thus, the composition can be in the form of, for example, a tablet, pill, powder, lozenge, sachet, cachet, elixir, suspension, emulsion, solution, syrup, aerosol (as a solid or in a liquid medium), ointment containing up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injection solutions, and sterile packaged powders. In some embodiments, the composition is formulated for oral administration. In some embodiments, the composition is a solid oral formulation. In some embodiments, the composition is formulated as a tablet or capsule.

[0234] Further provided herein is a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof together with a pharmaceutically acceptable carrier. Pharmaceutical compositions containing a compound of formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient can be prepared by intimately mixing the compound of formula (I) or a pharmaceutically acceptable salt thereof with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier can take a wide variety of forms depending on the desired route of administration (e.g., oral, parenteral). In some embodiments, the composition is a solid oral composition.

[0235] Suitable pharmaceutically acceptable carriers are well known in the art, and descriptions of some of these pharmaceutically acceptable carriers can be found in The Handbook of Pharmaceutical Excipients, published by the American Pharmaceutical Association and the British Pharmaceutical Society.

[0236] Methods for formulating pharmaceutical compositions are described in numerous publications, such as Pharmaceutical Dosage Forms: Tablets, Second Edition, Revised and Expanded, Volumes 1-3, edited by Lieberman et al., Pharmaceutical Dosage Forms: Parenteral Medications, Volumes 1-2, edited by Avis et al., and Pharmaceutical Dosage Forms: Disperse Systems, Volumes 1-2, edited by Lieberman et al. and published by Marcel Dekker, Inc.

[0237] When preparing compositions in oral dosage form, any of the usual pharmaceutical media can be used. Thus, for liquid oral preparations such as suspensions, elixirs, and solutions, suitable carriers and additives include water, glycols, oils, alcohols, flavoring agents, preservatives, stabilizers, coloring agents, etc.; for solid oral preparations such as powders, capsules, and tablets, suitable carriers and additives include starch, sugars, diluents, granulating agents, lubricants, binders, disintegrating agents, etc. Suitable binders include, but are not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth, or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, etc. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum, etc. Solid oral preparations can also be coated with a substance such as sugar to modify the primary absorption site, or enteric coated. For parenteral administration, the carrier usually consists of sterile water, and other ingredients may be added to increase solubility or preservation. Injection suspensions or solutions may also be prepared using aqueous carriers with appropriate additives. The pharmaceutical compositions herein will contain the amount of active ingredient necessary to deliver an effective dose as described herein per dosage unit, e.g., tablet, capsule, powder, injection, teaspoon, etc.

[0238] Compositions containing a compound of formula (I) or a pharmaceutically acceptable salt thereof can be formulated in unit dosage form, each dosage containing from about 1 to about 1,000 mg (1 g) of the active ingredient. The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for human and other subjects, each unit containing a predetermined amount of the active material (i.e., a compound of formula (I) or a pharmaceutically acceptable salt thereof) calculated to produce a desired therapeutic effect, in association with a suitable pharmaceutical excipient.

[0239] Active compound can be effective in a wide dosage range, and is generally administered in a pharmaceutically effective amount.The optimal dosage to be administered can be easily determined by those skilled in the art.Therefore, it will be understood that the amount of compound actually administered will usually be determined by a doctor and will vary according to relevant circumstances, including administration method, the actual compound to be administered, the strength of the preparation, the condition to be treated, and the progression of the disease state.In addition, factors related to the specific subject being treated, including subject's response, age, weight, diet, administration time, and the severity of subject's symptoms, will result in the need to adjust dosage.

[0240] Those skilled in the art will recognize that both in vivo and in vitro tests using suitable, known, and generally accepted cellular and / or animal models are predictive of the ability of a test compound to treat or prevent a given disorder.

[0241] Those skilled in the art will further recognize that human clinical trials, including first-in-human dose ranging and efficacy studies in healthy subjects and / or subjects afflicted with a given disorder, can be completed according to methods well known in the clinical and medical arts.

[0242] Provided herein are pharmaceutical kits useful for treating CDC7-associated diseases or disorders, such as cancer, comprising one or more containers containing a pharmaceutical composition comprising an effective amount of a compound provided herein. Such kits can, if desired, further comprise one or more of a variety of conventional pharmaceutical kit components, such as, for example, a container with one or more pharmaceutically acceptable carriers, additional containers, etc., as will be readily apparent to one of skill in the art. Instructions, either as an insert or label, indicating the amounts of components to be administered, guidelines for administration, and / or guidelines for mixing the components, can also be included in the kit. [Example]

[0243] Materials and Methods The compounds provided herein, including the salts thereof, can be prepared using known organic synthesis techniques and can be synthesized according to any of a number of possible synthetic routes.

[0244] The reactions for preparing the compounds provided herein can be carried out in a suitable solvent that can be easily selected by those skilled in the art of organic synthesis. A suitable solvent can be substantially non-reactive with the starting materials (reactants), intermediates, or products at the temperature at which the reaction is carried out, which can range, for example, from the freezing temperature of the solvent to the boiling temperature of the solvent. A given reaction can be carried out in one solvent or a mixture of two or more solvents. Depending on the specific reaction step, a suitable solvent for a particular reaction step can be selected by those skilled in the art.

[0245] Preparation of the compounds provided herein can involve the protection and deprotection of various chemical groups. The need for protection and deprotection, and the selection of appropriate protecting groups, can be readily determined by one of ordinary skill in the art. Protecting group chemistry is described, for example, in Protecting Group Chemistry, 1 st Ed.,Oxford University Press,2000,March's Advanced Organic Chemistry:Reactions,Mechanisms,and Structure,5 th Ed., Wiley-Interscience Publication, 2001, and Peturssion, S. et al., "Protecting Groups in Carbohydrate Chemistry," J. Chem. Educ., 74(11), 1297 (1997).

[0246] The reaction can be monitored according to any suitable method known in the art. For example, product formation can be monitored by nuclear magnetic resonance spectroscopy (e.g., 1 H or 13The activity of the compound may be monitored by spectroscopic means such as spectrophotometric methods, such as infrared spectroscopy, spectrophotometry (e.g., UV-visible light), mass spectroscopy, or by chromatographic methods such as high performance liquid chromatography (HPLC), liquid chromatography mass spectrometry (LCMS), or thin-layer chromatography (TLC). Compounds may be purified by those skilled in the art by a variety of methods, including high performance liquid chromatography (HPLC) ("Preparative LC-MS Purification: Improved Compound Specific Method Optimization" KF Blom, et al., J. Combi. Chem. 6(6), 874 (2004)), normal phase silica chromatography, and supercritical fluid chromatography (SFC).

[0247] Unless otherwise specified, the stereochemistry of compounds 1-58 in the following examples is understood to be arbitrarily assigned.

[0248] All solvents and reagents were obtained from commercial sources and used without further purification unless otherwise indicated. Anhydrous solvents were purchased and used as supplied. Reactions were monitored by thin-layer chromatography (TLC) visualized using a UV lamp (254 nm) and KMnO4 stain. NMR spectra were obtained on a Bruker Neo 400M spectrometer operating at 400 MHz. Chemical shifts are reported in parts per million (δ) from the tetramethysilane resonance in the indicated solvent. LC-mass spectra were acquired on an Agilent 1260-6125B single quadrupole mass spectrometer using a Welch Biomate column (C18, 2.7 μm, 4.6 × 50 mm) or a Waters H-Class SQD2 system. Detection was performed by DAD (254 nm, 210 nm, and 280 nm). Chiral HPLC was performed on a Waters Acquity UPC2 system using base-containing Daicel Chiralpak AD-H (5 μm, 4.6 × 250 mm), Daicel Chiralpak OD-H (5 μm, 4.6 × 250 mm), Daicel Chiralpak IG-3 (3 μm, 4.6 × 150 mm), Chiral Technologies Europe AD-3 (3 μm, 3.0 × 150 mm), and Trefoil Technology Trefoil AMY1 (2.5 μm, 3.0 × 150 mm). Detection was by DAD (254 nm). Preparative HPLC was performed on a Gilson Trilution LC system using a Welch XB-C18 column (5 μm, 21.2 × 150 mm). Flash chromatography was performed on a Biotage Isolera Prime system using Welch WelFlash flash columns (40-63 μm). All synthesized compounds had a purity of 95% or higher unless otherwise specified.

[0249] Abbreviation * = indicates that the amount of solvent or reagent before the "*" is used in the technique a number of times equal to the number after the "*". ℃=Celsius temperature 1H NMR = proton nuclear magnetic resonance spectrum AcOH = acetic acid Boc2O = tert-butoxycarbonyl anhydride con.=concentration d = doublet DCM = dichloromethane DIAD = diisopropyl azodicarboxylate DIPEA = N,N-diisopropylethylamine DMF = N,N-dimethylformamide EA = ethyl acetate ESI = electrospray ionization g = grams h=time HATU = (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, hexafluorophosphate azabenzotriazole tetramethyluronium HPLC = High-Performance Liquid Chromatography LCMS = Liquid Chromatography-Mass Spectrum M=mass m / z=mass-to-charge ratio MeCN = acetonitrile MeOH = methanol MeONa = sodium methoxide mg = milligram mL = milliliters mmol = millimolar mol = mole MS = mass spectrum NBS = N-bromosuccinimide obsd. = observed PCy3 = tricyclohexylphosphine Pd(AcO)2 = palladium(II) acetate Pd(dppf)Cl2 = (1,1'-bis(diphenylphosphino)ferrocene)palladium(II) dichloride PE = petroleum ether ppm=parts per million PTSA = para-toluenesulfonic acid rt=room temperature SFC = Supercritical Fluid Chromatography s = singlet t = triplet TBAF = tetrabutylammonium fluoride TFA = trifluoroacetic acid THF = tetrahydrofuran TLC = thin layer chromatography Trixiephos = rac-2-(di-tert-butylphosphino)-1,1'-binaphthyl

[0250] Example 1 - Compound 1: 2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one [ka] Step A: 6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one: [ka] To a solution of methyl 3-bromothiophene-2-carboxylate (1.00 g, 4.52 mmol, 1.0 equiv.), (5-diphenylphosphanyl-9,9-dimethyl-xanthen-4-yl)-diphenylphosphane (787 mg, 1.36 mmol, 0.3 equiv.), cesium carbonate (3.68 g, 11.30 mmol, 2.5 equiv.), sodium metabisulfite (172 mg, 0.90 mmol, 0.2 equiv.), and tris(dibenzylideneacetone)dipalladium (824 mg, 0.90 mmol, 0.20 equiv.) in toluene (40.0 mL) was added cyclohexanone (888 mg, 9.04 mmol, 2.0 equiv.) at room temperature. The reaction mixture was stirred at 105 °C under N for 16 h. The solvent was concentrated to dryness, and the residue was purified by flash chromatography (SiO, 0-10% EtOAc in PE) to give 6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (600 mg, 64%). MS obsd. (ESI + ):207.0[(M+H) + ].

[0251] Step B: 2-iodo-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one: [ka] To a solution of 6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (400 mg, 1.94 mmol, 1.0 equiv) in THF (15.0 mL) was added (diisopropylamino)lithium (2 M in THF, 1.1 mL, 2.2 mmol, 1.1 equiv) at −78° C. The mixture was stirred at −78° C. for 30 minutes, and then I (492 mg, 1.94 mmol, 1.0 equiv) was added. The mixture was stirred at −78° C. for 1 hour and then quenched with aqueous NaSO (10 mL) at 0° C. The mixture was extracted with DCM (10 mL × 3), and the combined organic phase was washed with water (20 mL), dried over anhydrous NaSO, filtered, and concentrated. The residue was purified by flash column chromatography (SiO, 0-10% EtOAc in PE) to give 2-iodo-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (300 mg, 46%). MS obsd. (ESI + ):333.1[(M+H) + ].

[0252] Step C: 2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one: [ka] 2-Iodo-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (300 mg, 903 μmol, 1.0 equiv.), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (175 mg, 900 μmol, 1.0 equiv.), sodium carbonate in water (2.0 mL) and 1,4-dioxane (10.0 mL). To a solution of 2,4,6-triisopropylphenylphenyl]phosphane (287.18 mg, 2.71 mmol, 3.0 equiv.) and dicyclohexyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane (129 mg, 270.96 μmol, 0.3 equiv.), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (132 mg, 181 μmol, 0.20 equiv.) was added at room temperature. The mixture was stirred at 110 °C in a microwave reactor for 2 h, then concentrated, and the residue was purified by flash column chromatography (SiO2, 0-50% EtOAc in PE) to give 2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (230 mg, 93%). MS obsd. (ESI + ):273.2[(M+H) + ].

[0253] Step D: 2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 1): [ka] To a solution of 2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (230 mg, 844.59 μmol, 1.0 equiv.) in methanol (2.0 mL) was added aqueous ammonia (6.0 mL, 28% w / w). The mixture was stirred in a sealed tube at 120° C. for 16 hours. After cooling at room temperature, the mixture was concentrated to dryness. The residue was dissolved in DMF and purified by preparative HPLC to give 2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (14 mg, 6.0%). MS obsd. (ESI + ):272.1[(M+H) + ]; 1 H NMR(400MHz,DMSO-d6)δ ppm:13.19(s,1H),11.15(s,1H),8.28(brs,1H),7.96(brs,1H),7.42(s,1H),2.67-2.62(m,2H),2.53-2.50(m,2H),1.75(s,4H).

[0254] Example 2 - Compound 2: 1,8,8-trifluoro-2-(1H-pyrazol-4-yl)-5,6,7,9-tetrahydrothieno[2,3-c]quinolin-4-one [ka] Step A: Methyl 3,5-dibromo-4-fluoro-thiophene-2-carboxylate: [ka] To a solution of methyl 4-fluorothiophene-2-carboxylate (1.00 g, 6.24 mmol, 1.0 equiv) in carbon tetrachloride (5.0 mL) was added FeBr (1.29 g, 4.37 mmol, 1.0 equiv) and bromine (39.9 g, 250 mmol, 20.0 mL, 40.0 equiv) at 25 °C, and the mixture was stirred for 16 h at 25 °C. The mixture was concentrated and purified by flash column chromatography (SiO, 0–15% EtOAc in PE) to give methyl 3,5-dibromo-4-fluoro-thiophene-2-carboxylate (480 mg, 24.0%). 1 H NMR(400MHz,CDCl3)δ ppm:3.91(s,3H).

[0255] Step B: Methyl 3-bromo-4-fluoro-5-[1-(2-trimethylsilylethoxymethyl)pyrazol-4-yl]thiophene-2-carboxylate: [ka] A solution of palladium(II) acetate (8 mg, 0.04 mmol, 0.05 equiv) and (5-diphenylphosphanyl-9,9-dimethyl-xanthen-4-yl)-diphenyl-phosphane (22 mg, 0.04 mmol, 0.05 equiv) in THF (10.0 mL) was stirred at 25 °C under N for 5 min. To the mixture were added methyl 3,5-dibromo-4-fluoro-thiophene-2-carboxylate (240 mg, 0.75 mmol, 1.0 equiv.), trimethyl-[2-[[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl]methoxy]ethyl]silane (367 mg, 1.13 mmol, 1.5 equiv.), and tripotassium orthophosphate (481 mg, 2.26 mmol, 3.0 equiv.), and the mixture was stirred at 60 °C under N for 16 h. The mixture was then concentrated in vacuo and purified by flash column chromatography (SiO, 0-30% EtOAc in PE) to give 3-bromo-4-fluoro-5-[1-(2-trimethylsilylethoxymethyl)pyrazol-4-yl]thiophene-2-carboxylate (168 mg, 51%). MS obsd. (ESI+): 79 Br / 81 Br 435.3 / 437.3 [(M+H) + ].

[0256] Step C: 1,8,8-trifluoro-2-[1-(2-trimethylsilylethoxymethyl)pyrazol-4-yl]-7,9-dihydro-6H-thieno[2,3-c]chromen-4-one [ka] To a solution of methyl 3-bromo-4-fluoro-5-[1-(2-trimethylsilylethoxymethyl)pyrazol-4-yl]thiophene-2-carboxylate (464 mg, 1.07 mmol, 1.0 equiv) in toluene (100.0 mL) was added tris(dibenzylideneacetone)dipalladium (98 mg, 0.11 mmol, 0.10 equiv), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (93 mg, 0.16 mmol, 0.15 equiv), cesium carbonate (1.0 g, 3.20 mmol, 3.0 equiv), sodium metabisulfite (20 mg, 0.11 mmol, 0.1 equiv), and 4,4-difluorocyclohexanone (429 mg, 3.20 mmol, 3.0 equiv). The mixture was purged with N2, then heated to 105 °C and stirred for 16 h. The mixture was concentrated to dryness, and the residue was purified by flash chromatography (SiO2, 0-30% EtOAc in PE) to give the product, 1,8,8-trifluoro-2-[1-(2-trimethylsilylethoxymethyl)pyrazol-4-yl]-7,9-dihydro-6H-thieno[2,3-c]chromen-4-one (217 mg, 44%). MS obsd. (ESI+): 457.3 [(M+H) + ].

[0257] Step D: 1,8,8-trifluoro-2-[1-(2-trimethylsilylethoxymethyl)pyrazol-4-yl]-5,6,7,9-tetrahydrothieno[2,3-c]quinolin-4-one: [ka] To a solution of 1,8,8-trifluoro-2-[1-(2-trimethylsilylethoxymethyl)pyrazol-4-yl]-7,9-dihydro-6H-thieno[2,3-c]chromen-4-one (110 mg, 0.24 mmol, 1.0 equiv.) in MeOH (5.0 mL) was added ammonium hydroxide (4.5 g, 128 mmol, 5.0 mL, 320 equiv.) at 25° C., and the mixture was stirred at 95° C. for 8 hours under microwave irradiation. The solution was cooled and concentrated to give 1,8,8-trifluoro-2-[1-(2-trimethylsilylethoxymethyl)pyrazol-4-yl]-5,6,7,9-tetrahydrothieno[2,3-c]quinolin-4-one (109 mg, crude), which was used in the next step without further purification. MS obsd. (ESI + ):456.3[(M+H) + ].

[0258] Step E: 1,8,8-trifluoro-2-(1H-pyrazol-4-yl)-5,6,7,9-tetrahydrothieno[2,3-c]quinolin-4-one (compound 2): [ka] To a solution of 1,8,8-trifluoro-2-[1-(2-trimethylsilylethoxymethyl)pyrazol-4-yl]-5,6,7,9-tetrahydrothieno[2,3-c]quinolin-4-one (109 mg, 0.24 mmol, 1.0 equiv) in DCM (1.0 mL) was added 2,2,2-trifluoroacetic acid (5.9 g, 52 mmol, 4.0 mL, 220 equiv) and the mixture was stirred at 25 °C for 1 h. The mixture was concentrated to dryness, and the residue was purified by flash column chromatography (SiO, 0–8% MeOH in DCM) to afford the impurity. The residue was further purified by reverse-phase column chromatography (C18 SiO2, 0-40% MeCN, 0.1% FA in water) to give 1,8,8-trifluoro-2-(1H-pyrazol-4-yl)-5,6,7,9-tetrahydrothieno[2,3-c]quinolin-4-one (22 mg, 28%). MS obsd. (ESI +):326.2[(M+H) + ].

[0259] Examples 3 and 4 - Compounds 3 and 4: (S)-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 3) and (R)-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 4) (The stereochemistry of Compounds 3 and 4 is arbitrarily assigned) [ka] Step A: 2-benzyloxycyclohexanol: [ka] BnOH (910 mg, 20 mmol, 2.0 equiv) was added dropwise to a stirred suspension of NaH (820 mg, 21.4 mmol, 60% in mineral oil, 2.0 equiv) in anhydrous DMF (25 mL) at 0 °C. After the mixture was stirred at 0 °C for 1 h, 7-oxabicyclo[4.1.0]heptane (1.0 g, 10 mmol, 1.0 equiv) was added. The mixture was heated at 60 °C for 2 h and then cooled. Water was added, and the mixture was extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous NaSO, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography (SiO, 0–50% EtOAc in PE) to give 2-benzyloxycyclohexanol (1.0 g, 48%). MS obsd. (ESI + ):207.3[(M+H) + ].

[0260] Step B: 2-benzyloxycyclohexanone: [ka] Oxalyl chloride (738 mg, 5.82 mmol, 1.2 equiv) was dissolved in anhydrous DCM (5.0 mL) under a nitrogen atmosphere and cooled to -78 °C. A solution of DMSO (909 mg, 11.6 mmol, 2.0 equiv) in anhydrous DCM (5.0 mL) was added dropwise, and the resulting mixture was stirred for 20 min. 2-Benzyloxycyclohexanol (1.0 g, 4.85 mmol, 1.0 equiv) in anhydrous DCM (5.0 mL) was added dropwise, forming a white precipitate. The mixture was stirred for 30 min, warmed to -60 °C, and anhydrous triethylamine (2.5 g, 24 mmol, 3.4 mL, 1.0 equiv) was added dropwise. The mixture was held at -60 °C for 5 min and then warmed to room temperature for 2 h. The reaction was quenched with HO and extracted with EtOAc. The organic layers were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography (SiO, 0-50% EtOAc in PE) to give 2-benzyloxycyclohexanone (770 mg, 78%). MS obsd. (ESI + ):205.2[(M+H) + ].

[0261] Step C: 6-(benzyloxy)-6,7,8,9-tetrahydrothieno[2,3-c]chromen-4-one: [ka] To a solution of methyl 3-bromothiophene-2-carboxylate (1.0 g, 4.5 mmol, 1.0 equiv.) in toluene (15 mL) was added 2-benzyloxycyclohexanone (1.4 g, 6.8 mmol, 1.5 equiv.), tris(dibenzylideneacetone)dipalladium (414 mg, 0.45 mmol, 0.1 equiv.), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (785 mg, 1.36 mmol, 0.30 equiv.), cesium carbonate (3.0 g, 9.1 mmol, 2.0 equiv.), and sodium metabisulfite (170 mg, 0.90 mmol, 0.20 equiv.). The reaction was purged with nitrogen and stirred at 105 °C for 3 h. The mixture was concentrated in vacuo, and the residue was purified by flash column chromatography (SiO, 0-50% EtOAc in PE) to give 6-(benzyloxy)-6,7,8,9-tetrahydrothieno[2,3-c]chromen-4-one (450 mg, 32%). MS obsd. (ESI + ):313.3[(M+H) + ].

[0262] Step D: 6-(benzyloxy)-2-iodo-6,7,8,9-tetrahydrothieno[2,3-c]chromen-4-one [ka] To a solution of 6-benzyloxy-6,7,8,9-tetrahydrothieno[2,3-c]chromen-4-one (350 mg, 1.12 mmol, 1.0 equiv) and I (569 mg, 2.24 mmol, 2.0 equiv) in THF (5.0 mL) was added LDA (2 M in THF, 2.2 mL, 4.0 equiv) at −65° C. The mixture was stirred at −65° C. for 1 h, then quenched with HO and extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo, and the residue was purified by flash column chromatography (SiO, 0–50% EtOAc in PE) to give 6-(benzyloxy)-2-iodo-6,7,8,9-tetrahydrothieno[2,3-c]chromen-4-one (300 mg, 61%). MS obsd. (ESI + ):439.3[(M+H) + ].

[0263] Step E: 6-(benzyloxy)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one: [ka] Trimethyl-[2-[[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl]methoxy]ethyl]silane (444 mg, 1.37 mmol, 1.2 equiv), 6-(benzyloxy)-2-iodo-6,7,8,9-tetrahydro- A suspension of thieno[2,3-c]chromen-4-one (500 mg, 1.10 mmol, 1.0 equiv), Pd(dppf)Cl (80 mg, 0.11 mmol, 0.1 equiv), X-Phos (104 mg, 0.22 mmol, 0.2 equiv), and CsCO (1.08 g, 3.30 mmol, 3.0 equiv) was irradiated in a microwave reactor at 110 °C for 2 h. The mixture was cooled and extracted with EtOAc. The organic layer was concentrated to dryness, and the residue was purified by flash column chromatography (SiO, 0–70% EtOAc in PE) to give 6-(benzyloxy)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (450 mg, 80%). MS obsd. (ESI + ):509.7[(M+H) + ].

[0264] Step F: 6-(benzyloxy)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one: [ka] A suspension of 6-(benzyloxy)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (500 mg, 1.32 mmol, 1.0 equiv) in a mixture of MeOH (7.0 mL) and 25% aqueous ammonia (7.0 mL) was irradiated in a microwave reactor at 100° C. for 3 hours. The resulting solid was filtered, washed with MeOH, and dried to give 6-(benzyloxy)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (250 mg, 50%). MS obsd. (ESI + ):508.7[(M+H) + ]

[0265] Step G: (S)-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 3) and (R)-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 4): [ka] A solution of 6-benzyloxy-2-[1-(2-trimethylsilylethoxymethyl)pyrazol-4-yl]-6,7,8,9-tetrahydro-5H-thieno[2,3-c]quinolin-4-one (250 mg, 0.49 mmol, 1.0 equiv.) in trifluoroacetic acid (10.0 mL) was heated at 70° C. for 1 h. The solvent was removed, and to the residue was added NH (7 M in MeOH, 5.0 mL). The mixture was stirred at room temperature for 5 min, concentrated, and the residue was purified by preparative HPLC to give 6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (60 mg, 40%). MS obsd. (ESI + ):288.4[(M+H) + ].

[0266] The racemic product was separated by chiral SFC to give the individual enantiomers. (S)-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 3): MS obsd. (ESI + ):288.2[(M+H) + ]. (R)-6-Hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 4): MS obsd. (ESI + ):288.2[(M+H) + ]; 1 H NMR(400MHz,DMSO-d6)δ ppm:13.20(s,1H),10.82(s,1H),8.30(s,1H),7.96(s,1H),7.47(s,1H),5.22(d,J=5.6Hz,1H), 4.49-4.46(m,1H),2.70-2.65(m,1H),2.51-2.49(m,1H),1.89-1.82(m,2H),1.78-1.69(m,2H).

[0267] Examples 5 and 6 - Compounds 5 and 6: (R)-6-(3,3-difluoropyrrolidin-1-yl)-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 5) and (S)-6-(3,3-difluoropyrrolidin-1-yl)-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 6). (The stereochemistry of Compounds 5 and 6 is arbitrarily assigned.) [ka] Step A: 6-Bromo-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one: [ka] A suspension of 6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-5H-thieno[2,3-c]quinolin-4-one (120 mg, 0.41 mmol, 1.0 equiv.) in TMSBr (8.0 mL) was stirred in a sealed tube at 80° C. for 6 h. The solvent was removed in vacuo, and the residue was used directly in the next step without further purification. MS obsd. (ESI + ):302.4[(M-Br+OCH3)+H) + ].

[0268] Step B: (R)-6-(3,3-difluoropyrrolidin-1-yl)-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 5) and (S)-6-(3,3-difluoropyrrolidin-1-yl)-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 6): [ka] A suspension of 3,3-difluoropyrrolidine hydrochloride (1.9 g, 14 mmol, 20 equiv.) and N,N-diisopropylethylamine (1.8 g, 13.88 mmol, 20 equiv.) in MeCN (10.0 mL) was stirred at room temperature for 2 hours. The solid was filtered off, and the filtrate was added to a sealed vial containing 6-bromo-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-5H-thieno[2,3-c]quinolin-4-one (200 mg, 0.69 mmol, 1.0 equiv.). NaI (103 mg, 0.69 mmol, 1.0 equiv.) was added to the vial. The mixture was stirred at room temperature for 16 h, then concentrated in vacuo, and the residue was purified by flash column chromatography (SiO, 0-100% EtOAc in PE) to give racemic 6-(3,3-difluoropyrrolidin-1-yl)-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (60 mg, 19%). MS obsd. (ESI + ):377.5[(M+H)+ ].

[0269] The individual enantiomers were separated via chiral SFC. (R)-6-(3,3-difluoropyrrolidin-1-yl)-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 5): MS obsd. (ESI + ):377.3[(M+H) + ]; 1 H NMR(400MHz,DMSO-d6)δ ppm:13.20(s,1H),10.53(s,1H),8.30(s,1H),7.96(s,1H),7.47(s,1H),3.73(t,J=5 .6Hz,1H),3.06-2.93(m,2H),2.87-2.78(m,2H),2.68-2.62(m,2H),2.30-2.22(m,2H) ), 1.98-1.95 (m, 1H), 1.84-1.77 (m, 2H), 1.74-1.71 (m, 1H). (S)-6-(3,3-difluoropyrrolidin-1-yl)-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 6): MS obsd. (ESI + ):377.3[(M+H) + ].

[0270] Examples 7 and 8 - Compounds 7 and 8 6-Methoxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-,9-ethanothieno[2,3-c]quinolin-4(5H)-one (Compound 7) and 6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-6,9-ethanothieno[2,3-c]quinolin-4(5H)-one (Compound 8). [ka] Step A: 6-Methoxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-6,9-ethanothieno[2,3-c]chromen-4-one: [ka] Cesium carbonate (230 mg, 720 μmol, 3.0 equiv.) and Sphos-Pd-G3 (37 mg, 48 μmol, 0.2 equiv.) were added to a solution of methyl 3-bromo-5-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)thiophene-2-carboxylate (100 mg, 240 μmol, 1.0 equiv.) and 1-methoxybicyclo[2.2.2]octan-2-one (74 mg, 480 μmol, 2.0 equiv., synthesized according to the procedure described in WO 2007 / 070201) in toluene (6.0 mL). The mixture was heated at 105° C. under nitrogen for 8 hours and then concentrated in vacuo. The resulting residue was purified by flash column chromatography (SiO, 0-25% EtOAc in PE) to give 6-methoxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-6,9-ethanothieno[2,3-c]chromen-4-one (32 mg, 24%, 82% purity). MS obsd. (ESI + ):459.4[(M+H) + ].

[0271] Step B: 6-Methoxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-6,9-ethanothieno[2,3-c]quinolin-4(5H)-one: [ka] To a solution of 6-methoxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-6,9-ethanothieno[2,3-c]chromen-4-one (20 mg, 44 μmol, 1.0 equiv) in isopropanol (3.0 mL) was added aqueous ammonia (17% w / w, 12.0 mL). The mixture was stirred at 60 °C for 16 h, then concentrated in vacuo and purified by flash column chromatography (0-5% MeOH in SiO2 DCM) to give 6-methoxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-6,9-ethanothieno[2,3-c]quinolin-4(5H)-one (10 mg, 24%, 48% purity). MS obsd. (ESI + ):458.4[(M+H) + ].

[0272] Step C: 6-Methoxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-6,9-ethanothieno[2,3-c]quinolin-4(5H)-one (compound 7): [ka] A solution of 6-methoxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-6,9-ethanothieno[2,3-c]quinolin-4(5H)-one (28 mg, approximately 50% pure, 29 μmol, 1.0 equiv.) in DCM (4.0 mL) was cooled to 0° C., followed by the addition of trifluoroacetic acid (1.0 mL). The mixture was stirred at room temperature for 2 hours, then concentrated in vacuo and purified by preparative HPLC to give 6-methoxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-6,9-ethanothieno[2,3-c]quinolin-4(5H)-one (compound 7, 5 mg, 51%). MS obsd. (ESI + ):328.1[(M+H) + ]; 1H NMR(400MHz,DMSO-d6)δ ppm:13.16(s,1H),10.52(s,1H),8.10(s,2H),7.59(s,1H),3.39(s,3H),3.40-3.92(m,1 H),2.12(t,J=10.4Hz,2H),1.88(t,J=10.4Hz,2H),1.49-1.44(m,2H),1.36-1.31(m,2H).

[0273] Step D: 6-Hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-6,9-ethanothieno[2,3-c]quinolin-4(5H)-one (compound 8): [ka] A solution of 6-methoxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-6,9-ethanothieno[2,3-c]quinolin-4(5H)-one (55 mg, 120 μmol, 1.0 equiv.) in HBr (40% aqueous, 5.0 mL) was heated to 100° C. for 8 h. The mixture was concentrated in vacuo and purified by reverse-phase column chromatography (C18 SiO, 0-40% MeCN in water, 0.1% NH4HCO3 in water) to give 6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-6,9-ethanothieno[2,3-c]quinolin-4(5H)-one (Example 8, 16.2 mg, 31%). MS obsd. (ESI + ):314.2[(M+H) + ]; 1 H NMR(400MHz,DMSO-d6)δ ppm:13.17(s,1H),10.22(s,1H),8.14(brs,2H),7.58(s,1H),5.72(s,1H),3.30(s,1H),1.84-1.80(m,4H),1.43(d,J=6.4Hz,4H).

[0274] Example 9 - Compound 9: 6-hydroxy-2-(pyridin-4-yl)-6,7,8,9-tetrahydro-6,9-ethanothieno[2,3-c]quinolin-4(5H)-one [ka] Step A: Methyl 3-bromo-5-(pyridin-4-yl)thiophene-2-carboxylate: [ka] Cesium carbonate (210 mg, 680 μmol, 2.0 equiv.), Pd(dppf)Cl (240 mg, 330 μmol, 1.0 equiv.), and dicyclohexyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane (160 mg, 330 μmol, 1.0 equiv.) were added to a solution of methyl 3,5-dibromothiophene-2-carboxylate (100 mg, 330 μmol, 1.0 equiv.) and 4-pyridylboronic acid (45 mg, 370 μmol, 1.1 equiv.) in 1,4-dioxane (10.0 mL). The mixture was degassed twice with N and then heated at 80 °C for 1 h. The mixture was concentrated in vacuo and then purified by column chromatography (SiO, 0-25% EtOAc in PE) to give methyl 3-bromo-5-(pyridin-4-yl)thiophene-2-carboxylate (65 mg, 56%). MS obsd. (ESI + ):298.0[(M+H) + ].

[0275] Step B: 6-Methoxy-2-(pyridin-4-yl)-6,7,8,9-tetrahydro-4H-6,9-ethanothieno[2,3-c]chromen-4-one: [ka] Cesium carbonate (330 mg, 1.0 mmol, 3.0 equiv) and Sphos-Pd-G3 (52 mg, 67 μmol, 0.2 equiv) were added to a solution of methyl 3-bromo-5-(4-pyridyl)thiophene-2-carboxylate (100 mg, 340 μmol, 1.0 equiv) and 1-methoxybicyclo[2.2.2]octan-2-one (100 mg, 670 μmol, 2.0 equiv) in toluene (4.0 mL). The mixture was stirred under nitrogen at 100 °C for 16 h, concentrated in vacuo, and purified by flash column chromatography (SiO, 0-10% MeOH in DCM) to give 6-methoxy-2-(pyridin-4-yl)-6,7,8,9-tetrahydro-4H-6,9-ethanothieno[2,3-c]chromen-4-one (21 mg, 16%, 82% purity). MS obsd. (ESI + ):340.4[(M+H) + ].

[0276] Step C: 6-Methoxy-2-(pyridin-4-yl)-6,7,8,9-tetrahydro-4H-6,9-ethanothieno[2,3-c]chromen-4-one: [ka] To a solution of 6-methoxy-2-(pyridin-4-yl)-6,7,8,9-tetrahydro-4H-6,9-ethanothieno[2,3-c]chromen-4-one (15 mg, 44.19 μmol, 1.0 equiv.) in isopropanol (4.0 mL) was added aqueous ammonia (25% w / w, 16.0 mL). The mixture was stirred at 60° C. for 16 hours and then concentrated in vacuo to give 6-methoxy-2-(pyridin-4-yl)-6,7,8,9-tetrahydro-4H-6,9-ethanothieno[2,3-c]chromen-4-one (15 mg, crude), which was used without further purification. MS obsd. (ESI + ):357.3[(M+H) + ].

[0277] Step D: 6-Methoxy-2-(pyridin-4-yl)-6,7,8,9-tetrahydro-6,9-ethanothieno[2,3-c]quinolin-4(5H)-one: [ka] Crude 3-(4-methoxy-3-oxobicyclo[2.2.2]octan-2-yl)-5-(pyridin-4-yl)thiophene-2-carboxamide (30 mg) was dissolved in toluene (4.0 mL) and p-toluenesulfonic acid (3 mg, 17 μmol, 0.20 equiv.). The mixture was stirred at 100° C. for 1 h, after which NaHCO (saturated aqueous solution, 10 mL) was added. The mixture was extracted with EtOAc (3×15 mL), and the combined organic layers were dried over NaSO and filtered. Concentration in vacuo afforded 6-methoxy-2-(pyridin-4-yl)-6,7,8,9-tetrahydro-6,9-ethanothieno[2,3-c]quinolin-4(5H)-one (28 mg, crude), which was used without further purification. MS obsd. (ESI + ):339.2[(M+H) + ].

[0278] Step E: 6-hydroxy-2-(pyridin-4-yl)-6,7,8,9-tetrahydro-6,9-ethanothieno[2,3-c]quinolin-4(5H)-one (compound 9): [ka] A solution of 6-methoxy-2-(pyridin-4-yl)-6,7,8,9-tetrahydro-6,9-ethanothieno[2,3-c]quinolin-4(5H)-one (30 mg, 89 μmol) in HBr (48% aqueous solution, 4.0 mL) was heated to 100° C. for 16 hours, concentrated in vacuo, and purified by preparative HPLC to give 6-hydroxy-2-(pyridin-4-yl)-6,7,8,9-tetrahydro-6,9-ethanothieno[2,3-c]quinolin-4(5H)-one (compound 9, 11 mg, 38%). MS obsd. (ESI + ):325.1[(M+H) +]; 1 H NMR(400MHz,DMSO-d6)δ ppm:10.59(s,1H),8.68-8.67(m,2H),8.23(s,1H),7.82-7.81(m,2H),5.76(s,1H),3.43-3.41(m,1H),1.85-1.78(m,4H),1.46-1.43(m,4H).

[0279] Examples 10 and 11 - Compounds 10 and 11: (R)-4-methoxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 10) and (S)-4-methoxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 11) (stereochemistry arbitrarily assigned) [ka] Step A: 4-Methoxy-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one [ka] To a solution of methyl 3-bromo-5-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)thiophene-2-carboxylate (300 mg, 0.72 mmol, 1.0 equiv., prepared in a manner similar to Step A of Compound 9) and 3-methoxytetrahydropyran-4-one (190 mg, 1.4 mmol, 2.0 equiv., synthesis described in WO 2005 / 014537) in toluene (7.0 mL) was added sodium metabisulfite (14 mg, 72 μmol, 0.1 equiv.), CsCO (700 mg, 2.2 mmol, 3.0 equiv.), Pd(dba) (130 mg, 0.14 mmol, 0.2 equiv.), and Xantphos (170 mg, 0.29 mmol, 0.4 equiv.). The mixture was stirred at 100 °C for 16 h, concentrated, and purified by column chromatography (SiO, 0-25% EtOAc in PE) to give 4-methoxy-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (160 mg, 51%). MS obsd. (ESI + ):435.4[(M+H) + ].

[0280] Step B: 4-Methoxy-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] A solution of 4-methoxy-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (160 mg, 0.37 mmol, 1.0 equiv) in 1:4 trifluoroacetic acid / DCM (10 mL) was stirred at room temperature for 2 h. The mixture was concentrated to give 4-methoxy-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (112 mg, crude), which was used without further purification. MS obsd. (ESI + ):305.2[(M+H) + ].

[0281] Step C: (R)-4-Methoxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 10) and (S)-4-Methoxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 11): [ka] A solution of 4-methoxy-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (110 mg, 1.0 equiv.) in 10 mL of NH4OH / MeOH (1 / 1) was stirred in a microwave reactor at 100 °C for 6 h. The mixture was concentrated in vacuo and purified by preparative HPLC to give racemic 4-methoxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (75 mg, 77%). MS obsd. (ESI + ):304.2[(M+H) + ].

[0282] The individual enantiomers were separated by chiral SFC. (R)-4-Methoxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 10): MS obsd. (ESI + ):304.2[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d6) δ ppm: 13.23 (s, 1H), 11.53 (s, 1H), 8.30 (brs, 2H), 7.44 (s, 1H), 4.88-4.69 (m, 1H), 4.64-4.46 (m, 1H), 4.18 (d, J = 12.0 Hz, 1H), 4.01 (s, 1H), 3.70 (d, J = 12.0 Hz, 1H), 3.39 (s, 3H). (S)-4-Methoxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 11): MS obsd. (ESI + ):304.2[(M+H) + ].

[0283] Examples 12 and 13—Compounds 12 and 13: (S)-6-(azetidin-1-yl)-8,8-difluoro-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 12) and (R)-6-(azetidin-1-yl)-8,8-difluoro-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 13) (stereochemistry arbitrarily assigned) [ka] Step A: 5,5-Difluoro-2,2-dimethoxy-cyclohexanol: [ka] 4,4-Difluorocyclohexanone (4.0 g, 30 mmol, 1.0 equiv) and KOH (4.0 g, 72 mmol, 3.0 equiv) were dissolved in MeOH (80.0 mL). The mixture was cooled to 0 °C, and a solution of I2 (8.3 g, 33 mmol, 1.1 equiv in 100.0 mL of MeOH) was added dropwise over 1 h. The reaction was stirred at room temperature for 18 h, concentrated in vacuo, and the oil was suspended in 80 mL of DCM. The precipitate was filtered off, and the solution was concentrated to give crude 5,5-difluoro-2,2-dimethoxy-cyclohexanol (4.2 g, crude).

[0284] Step B: (((5,5-Difluoro-2,2-dimethoxycyclohexyl)oxy)methyl)benzene [ka] Sodium hydride (610 mg, 15 mmol, 60% in mineral oil, 1.2 equiv.) was added to a solution of BnBr (2.8 g, 17 mmol, 1.3 equiv.) in DMF (30.0 mL) at 0 °C. After 30 min, 5,5-difluoro-2,2-dimethoxy-cyclohexanol (2.5 g crude, 13 mmol, 1.0 equiv.) was added, and the mixture was stirred at room temperature for 16 h. The mixture was poured into water and extracted with EtOAc (15 mL × 2). The organic layer was dried, filtered, concentrated, and purified by column chromatography (SiO, 0–12% EtOAc in PE) to give (((5,5-difluoro-2,2-dimethoxycyclohexyl)oxy)methyl)benzene (3.2 g, 64%). 1 H NMR(400MHz,CDCl3)δ ppm:7.37-7.25(m,5H),4.72(d,J=12.0Hz,1H),4.56(d,J=12.0Hz,1H),3.70(t ,J=8.0Hz,1H),3.24(s,3H),3.22(s,3H),2.38-2.35(m,1H),2.11-1.74(m,5H).

[0285] Step C: 2-(benzyloxy)-4,4-difluorocyclohexan-1-one: [ka] To a solution of (((5,5-difluoro-2,2-dimethoxycyclohexyl)oxy)methyl)benzene (2.2 g, 7.7 mmol, 1.0 equiv.) in acetone (60.0 mL) was added I2 (195 mg, 768 μmol, 0.1 equiv.). After 30 min, Na2SO3 (saturated aqueous solution) was added, and the aqueous phase was extracted with DCM (15 mL × 3). The combined organic phases were washed with Na2SO3 (saturated aqueous solution), filtered, and concentrated. Purification by column chromatography (SiO2, 0–8% EtOAc in PE) gave 2-(benzyloxy)-4,4-difluorocyclohexan-1-one (1.3 g, 70%). 1 H NMR (400MHz, CDCl3): δ ppm:7.36-7.25(m,5H),4.85(d,J=11.2Hz,1H),4.51(d,J=11.2Hz,1H),4.15-4.11(m,1H),2.75-2.14(m,6H).

[0286] Step D: 6-(benzyloxy)-8,8-difluoro-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one: [ka] To a solution of methyl 3-bromo-5-[1-(2-trimethylsilylethoxymethyl)pyrazol-4-yl]thiophene-2-carboxylate (400 mg, 960 μmol, 1.0 equiv.) and 2-(benzyloxy)-4,4-difluorocyclohexan-1-one (460 mg, 1.9 mmol, 2.0 equiv.) in toluene (80.0 mL) was added CsCO (940 mg, 2.9 mmol, 3.0 equiv.), Pd(dba) (180 mg, 190 μmol, 0.2 equiv.), Xantphos (170 mg, 270 μmol, 0.3 equiv.), and NaSO (54 mg, 290 μmol, 0.3 equiv.). The mixture was stirred and heated at 105 °C for 16 h. The mixture was cooled, filtered, concentrated, and the residue was purified by silica gel chromatography (SiO, 0-28% EtOAc in PE) to give 6-(benzyloxy)-8,8-difluoro-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (360 mg, 55%). MS obsd. (ESI + ):545.3[(M+H) + ].

[0287] Step E: 6-(benzyloxy)-8,8-difluoro-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one: [ka] To a solution of 6-(benzyloxy)-8,8-difluoro-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (150 mg, 200 μmol, 1.0 equiv.) in MeOH (8.0 mL) was added aqueous ammonia (30% w / w, 8.0 mL). The mixture was heated in a microwave reactor at 95° C. for 8 hours and then cooled. The mixture was extracted with DCM (10 mL×3), and the combined organic phases were dried over sodium sulfate, filtered, and concentrated. Purification by column chromatography (SiO, 0-4% MeOH in DCM) gave 6-(benzyloxy)-8,8-difluoro-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (90 mg, 69%). MS obsd. (ESI + ):544.4[(M+H) + ].

[0288] Step F: 8,8-Difluoro-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one: [ka] A solution of 6-(benzyloxy)-8,8-difluoro-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (200 mg, 367 μmol, 1.0 equiv) in DCM (12.0 mL) was cooled to 0 °C, and BCl (1 M in n-hexane, 3.68 mL, 3.68 mmol, 10 equiv) was added. After 1 h at 0 °C, MeOH (3.0 mL) was added, and the solution was concentrated to give a solid. Trituration (1:1 n-hexane / MeOH) gave 8,8-difluoro-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (110 mg, 92% yield) of sufficient purity for the following step. MS obsd. (ESI + ):324.1[(M+H) + ].

[0289] Step G: (R)-6-(azetidin-1-yl)-8,8-difluoro-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 12) and (S)-6-(azetidin-1-yl)-8,8-difluoro-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 13): [ka] To 8,8-difluoro-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (70 mg, 190 μmol, 1.0 equiv.), SOCl (2.5 g, 21 mmol, 1.5 mL, 11 equiv.) was slowly added at room temperature. After 4 h, the mixture was concentrated. The concentrate containing 6-chloro-8,8-difluoro-2-(1H-pyrazol-4-yl)-5,6,7,9-tetrahydrothieno[2,3-c]quinolin-4-one was dissolved in MeCN (3.0 mL), and KI (241 mg, 1.45 mmol, 10.0 equiv.) and azetidine (170 mg, 2.9 mmol, 20 equiv.) were added. The mixture was stirred at room temperature for 12 hours, then concentrated and purified via reverse phase chromatography to give 6-(azetidin-1-yl)-8,8-difluoro-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (34 mg, 55%, 85% purity). MS obsd. (ESI + ):363.3[(M+H) + ].

[0290] The individual enantiomers were separated via chiral SFC. (R)-6-(azetidin-1-yl)-8,8-difluoro-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 12): MS obsd. (ESI + ):363.4[(M+H) + ]. (S)-6-(azetidin-1-yl)-8,8-difluoro-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 13): MS obsd. (ESI + ):363.3[(M+H) + ].

[0291] Examples 14 and 15 - Compounds 14 and 15: (S)-8,8-difluoro-6-(hydroxymethyl)-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 14) and (R)-8,8-difluoro-6-(hydroxymethyl)-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 15) (stereochemistry arbitrarily assigned) [ka] Step A: Methyl 5,5-difluoro-2-hydroxycyclohex-1-ene-1-carboxylate: [ka] A solution of 4,4-difluorocyclohexanone (2.0 g, 15 mmol, 1.0 equiv) in DMF (4.0 mL) was added to a suspension of NaH (430 mg, 18 mmol, 60% in mineral oil, 1.2 equiv) in DMF (13.0 mL) at 0 °C. After 30 min at 0 °C, a solution of dimethyl carbonate (1.6 g, 18 mmol, 1.5 mL, 1.2 equiv) in DMF (3.0 mL) was added, and the mixture was stirred at room temperature for 16 h, followed by the addition of NH Cl (saturated aqueous solution). The mixture was diluted with water (10 mL) and extracted with DCM (20 mL × 3). The combined organic layers were dried (Na SO ), concentrated in vacuo, and purified by column chromatography (SiO , 0–8% EtOAc in PE) to give methyl 5,5-difluoro-2-hydroxycyclohex-1-ene-1-carboxylate (2.0 g, 70%). 1 H NMR(400MHz,CDCl3)δ ppm:12.17(s,1H),3.78(s,3H),2.74(t,J=14.4Hz,2H),2.55(t,J=6.8Hz,2H),2.17-2.07(m,2H).

[0292] Step B: Methyl 5,5-difluoro-1-methyl-2-oxocyclohexane-1-carboxylate: [ka] To a flask containing methyl 5,5-difluoro-2-hydroxycyclohex-1-ene-1-carboxylate (1.0 g, 6.4 mmol, 1.0 equiv.) in acetone (12.0 mL), K2CO3 (2.7 g, 19 mmol, 3.0 equiv.) and iodomethane (2.7 g, 19 mmol, 3.0 equiv.) were added at room temperature. The mixture was heated to 50 °C and stirred for 3 h, after which water was added. The mixture was extracted with t-butyl methyl ether (30 mL × 3), and the combined organic layers were washed with water, brine, and dried (Na2SO4). Filtration, concentration in vacuo, and purification by column chromatography (SiO2, 0–15% EtOAc in PE) gave methyl 5,5-difluoro-1-methyl-2-oxocyclohexane-1-carboxylate (800 mg, 66%). 1 H NMR(400MHz,CDCl3)δ ppm:3.76(s,3H),3.09-2.88(m,2H),2.61-2.54(m,1H),2.46-2.35(m,1H),2.26-1.98(m,2H),1.37(d,J=0.8Hz,3H).

[0293] Step C: Methyl 8,8-difluoro-6-methyl-4-oxo-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromene-6-carboxylate: [ka] To a solution of methyl 3-iodo-5-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)thiophene-2-carboxylate (200 mg, 430 μmol, 1.0 equiv.) and methyl 5,5-difluoro-1-methyl-2-oxocyclohexane-1-carboxylate (180 mg, 860 μmol, 2.0 equiv.) in toluene (12.0 mL) was added CsCO (420 mg, 1.3 mmol, 3.0 equiv.), Pd(dba) (79 mg, 86 μmol, 0.2 equiv.), Xantphos (74 mg, 130 μmol, 0.30 equiv.), and NaSO (16 mg, 86 μmol, 0.2 equiv.). The mixture was heated to 105 °C in a microwave reactor for 2 h, then cooled, concentrated, and purified (SiO2, 0-32% EtOAc in PE). This material was further purified by reverse-phase chromatography to give methyl 8,8-difluoro-6-methyl-4-oxo-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromene-6-carboxylate (100 mg, 46%). MS obsd. (ESI + ):511.4[(M+H) + ].

[0294] Step D: 8,8-Difluoro-6-(hydroxymethyl)-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one: [ka] To a cooled (0 °C) solution of methyl 8,8-difluoro-6-methyl-4-oxo-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromene-6-carboxylate (100 mg, 200 μmol, 1.0 equiv.) in EtOH (12.0 mL) was added CaCl (45 mg, 410 μmol, 2.0 equiv.). After 10 min, NaBH (116 mg, 3.08 mmol, 15.0 equiv.) was added and the ice bath was removed. The mixture was stirred at room temperature for 20 min, then cooled to 0 °C, and NH Cl (sat. aq.) was added. The mixture was extracted with DCM (10 mL × 3), and the combined organic phases were dried (Na SO ), filtered, and concentrated. Purification by column chromatography (0-70% EtOAc on SiO2PE) gave 8,8-difluoro-6-(hydroxymethyl)-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (55 mg, 48%). MS obsd. (ESI + ):483.4[(M+H) + ].

[0295] Step E: 8,8-difluoro-6-(hydroxymethyl)-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one: [ka] To a solution of 8,8-difluoro-6-(hydroxymethyl)-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (98 mg, 200 μmol, 1.0 equiv.) in MeOH (5.0 mL) was added aqueous ammonia (5.0 mL, 30% w / w). The mixture was heated to 95° C. in a microwave reactor for 8 h. The mixture was concentrated and purified by column chromatography (0-7% MeOH in SiO2 DCM) to give 8,8-difluoro-6-(hydroxymethyl)-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (68 mg, 56%). MS obsd. (ESI + ):482.4[(M+H) + ].

[0296] Step F: (S)-8,8-difluoro-6-(hydroxymethyl)-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 14) and (R)-8,8-difluoro-6-(hydroxymethyl)-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 15): [ka] To a cooled (0° C.) solution of 8,8-difluoro-6-(hydroxymethyl)-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (43 mg, 89 μmol) in DCM (4.0 mL) was added trifluoroacetic acid (2.0 mL). The mixture was warmed to room temperature and stirred for 2 hours, then concentrated in vacuo and purified by reverse-phase chromatography to give 8,8-difluoro-6-(hydroxymethyl)-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (22 mg, 69%). MS obsd. (ESI + ):352.0[(M+H) + ]; 1 H NMR(400MHz,DMSO-d6):δ ppm 13.22(s,1H),10.86(s,1H),8.28(s,1H),7.98(s,1H),7.53(s,1H),5.23(s,1H),3.84(d,J=10.8Hz ,1H),3.41(d,J=10.4Hz,1H),3.29-3.13(m,2H),2.58-2.46(m,1H),2.03-1.94(m,1H),1.30(s,3H).

[0297] The individual enantiomers were separated by chiral SFC to give the respective enantiomers: (S)-8,8-difluoro-6-(hydroxymethyl)-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 14): MS obsd. (ESI + ):352.1[(M+H) + ]. (R)-8,8-Difluoro-6-(hydroxymethyl)-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 15): MS obsd. (ESI + ):352.1[(M+H) + ].

[0298] Examples 16 and 17 - Compounds 16 and 17: (S)-4-hydroxy-4-isopropyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 16) and (R)-4-hydroxy-4-isopropyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 17) [ka] Step A: 4-(benzyloxy)-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] To a solution of methyl 3-bromo-5-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)thiophene-2-carboxylate (5.0 g, 12 mmol, 1.0 equiv.) and 3-(benzyloxy)tetrahydro-4H-pyran-4-one (4.9 g, 24 mmol, 2.0 equiv.) in 1,4-dioxane (600 mL) was added NaSO (228 mg, 1.20 mmol, 0.1 equiv.), CsCO (12 g, 36 mmol, 3.0 equiv.), Pd(dba) (2.19 g, 2.40 mmol, 0.20 equiv.), and Xantphos (2.8 g, 4.8 mmol, 0.40 equiv.). The mixture was degassed twice with N2, then stirred and heated at 100 °C for 16 h. The mixture was filtered, concentrated in vacuo, and purified by flash column chromatography (SiO2, 0-60% EtOAc in PE) to give 4-(benzyloxy)-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (3.5 g, 38% yield, 81% purity). MS obsd. (ESI +):511.5[(M+H) + ].

[0299] Step B: 4-(benzyloxy)-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one: [ka] A solution of 4-(benzyloxy)-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (1.00 g, 1.6 mmol, 81% purity, 1.0 equiv) and NH4OH (6 mL) in MeOH (6.0 mL) was stirred at 100 °C for 6 h under microwave heating. The mixture was concentrated in vacuo to give a residue that was purified by flash column chromatography (SiO, 0-10% MeOH in DCM) to give 4-(benzyloxy)-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (560 mg, 68%). MS obsd. (ESI + ):510.2[(M+H) + ].

[0300] Step C: 4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one: [ka] To a solution of 4-(benzyloxy)-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (300 mg, 590 μmol, 1.0 equiv.) in DCM (2.0 mL) and BC1 (1 M in hexanes, 5.9 mL, 10 equiv.) was added. The mixture was stirred at 0° C. for 15 min and then concentrated in vacuo to give 4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (170 mg, crude), which was used in the next reaction without further purification. MS obsd. (ESI + ):290.0[(M+H) + ].

[0301] Step D: Tert-butyl 4-(4-hydroxy-6-oxo-3,4,5,6-tetrahydro-1H-pyrano[4,3-b]thieno[3,2-d]pyridin-8-yl)-1H-pyrazole-1-carboxylate: [ka] To a suspension of 4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (170 mg, crude) in DMF (2.0 mL) was added trimethylamine (119 mg, 1.18 mmol, 2.0 equiv), DMAP (36 mg, 290 μmol, 0.5 equiv), and (Boc)O (130 mg, 590 μmol, 1.0 equiv). The mixture was stirred at room temperature for 10 min, then poured into water and extracted with EtOAc / THF (10:1, 15 mL × 2). The combined organic layers were dried, filtered, concentrated, and purified by flash column chromatography (SiO, 0-10% MeOH in DCM) to give tert-butyl 4-(4-hydroxy-6-oxo-3,4,5,6-tetrahydro-1H-pyrano[4,3-b]thieno[3,2-d]pyridin-8-yl)-1H-pyrazole-1-carboxylate (130 mg, 56% over two steps). MS obsd. (ESI + ):390.0[(M+H) + ].

[0302] Step E: Tert-butyl 4-(4,6-dioxo-3,4,5,6-tetrahydro-1H-pyrano[4,3-b]thieno[3,2-d]pyridin-8-yl)-1H-pyrazole-1-carboxylate: [ka] Dess-Martin periodinane (425 mg, 1.00 mmol, 3.0 equiv) was added to a solution of tert-butyl 4-(4-hydroxy-6-oxo-3,4,5,6-tetrahydro-1H-pyrano[4,3-b]thieno[3,2-d]pyridin-8-yl)-1H-pyrazole-1-carboxylate (130 mg, 334 μmol, 1.0 equiv) in DCM (3.0 mL) and DMF (3.0 mL) at 0° C. The mixture was stirred at room temperature for 2 h, then saturated NaSO (aq) and NaHCO (aq) were added along with water (10 mL). The mixture was extracted with DCM / MeOH (10:1, 20 mL × 3), and the combined organic layers were dried, filtered, concentrated, and purified by flash column chromatography (SiO2, 0-10% MeOH in DCM) to give tert-butyl 4-(4,6-dioxo-3,4,5,6-tetrahydro-1H-pyrano[4,3-b]thieno[3,2-d]pyridin-8-yl)-1H-pyrazole-1-carboxylate (70 mg, 54%). MS obsd. (ESI + ):388.0[(M+H) + ].

[0303] Step F: (S)-4-hydroxy-4-isopropyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 16) and (R)-4-hydroxy-4-isopropyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 17): [ka] To a solution of tert-butyl 4-(4,6-dioxo-3,4,5,6-tetrahydro-1H-pyrano[4,3-b]thieno[3,2-d]pyridin-8-yl)-1H-pyrazole-1-carboxylate (1.96 g, 5.06 mmol, 1.0 equiv.) in anhydrous THF (6.0 mL) was added i-PrMgCl-LiCl (1.3 M in THF, 80 mL, 20 equiv.). The mixture was stirred at 0° C. for 30 min, then quenched with saturated aqueous NH4Cl and extracted with EtOAc / THF (10:1, 30 mL×2). The combined organic layers were dried, filtered, concentrated, and purified by flash column chromatography (SiO, 0-10% MeOH in DCM) to give racemic 4-hydroxy-4-isopropyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (680 mg, 41%). MS obsd. (ESI + ):332.0[(M+H) + ].

[0304] The individual enantiomers were separated via chiral SFC. (S)-4-Hydroxy-4-isopropyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 16): MS obsd. (ESI + ):332.0[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d): δ ppm: 13.22 (s, 1H), 10.81 (s, 1H), 8.29 (s, 1H), 7.96 (s, 1H), 7.41 (s, 1H), 5.25 (s, 1H), 4.67 (s, 2H), 3.96 (d, J = 11.6 Hz, 1H), 3.53 (d, J = 11.6 Hz, 1H), 2.41-2.34 (m, 1H), 1.00 (d, J = 6.8 Hz, 3H), 0.76 (d, J = 6.8 Hz, 3H). (R)-4-Hydroxy-4-isopropyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 17): MS obsd. (ESI+ ):332.0[(M+H) + ].

[0305] Examples 18 and 19 - Compounds 18 and 19: (S)-4-ethyl-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 18) and (R)-4-ethyl-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 19) [ka] To a solution of tert-butyl 4-(4,6-dioxo-3,4,5,6-tetrahydro-1H-pyrano[4,3-b]thieno[3,2-d]pyridin-8-yl)-1H-pyrazole-1-carboxylate (180 mg, 460 μmol) in anhydrous THF (2.0 mL) was added EtMgCl (2 M in THF, 12.0 mL, 52 equiv.) and the mixture was stirred for 1 h at 0° C. The mixture was quenched with saturated NH4Cl (aq.) and extracted with EtOAc:THF (10:1, 30 mL × 2). The combined organic layers were dried, filtered, concentrated, and purified by flash column chromatography (SiO, 0-10% MeOH in DCM) to give racemic 4-ethyl-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (60 mg, 41%). MS obsd. (ESI + ):318.1[(M+H) + ].

[0306] The individual enantiomers were separated via chiral SFC. (S)-4-Ethyl-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 18): MS obsd. (ESI + ):318.0[(M+H)+ ]; 1 H NMR (400 MHz, DMSO-d6) δ ppm: 13.22 (s, 1H), 10.88 (s, 1H), 8.29 (s, 1H), 7.95 (s, 1H), 7.40 (s, 1H), 5.28 (s, 1H), 4.86-4.49 (m, 2H), 3.85 (d, J = 11.2 Hz, 1H), 3.54 (d, J = 11.2 Hz, 1H), 2.00-1.79 (m, 2H), 0.85 (t, J = 7.6 Hz, 3H). (R)-4-Ethyl-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 19): MS obsd. (ESI + ):318.0[(M+H) + ].

[0307] Examples 20 and 21 - Compounds 20 and 21: (S)-4-(tert-butyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-3,4-dihydro-1H-pyrano[4,3-b]thieno[3,2-d]pyridin-6(5H)-one (Compound 20) and (R)-4-(tert-butyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-3,4-dihydro-1H-pyrano[4,3-b]thieno[3,2-d]pyridin-6(5H)-one (Compound 21). (The stereochemistry for Compounds 20 and 21 is arbitrarily assigned.) [ka] To a solution of tert-butyl 4-(4,6-dioxo-3,4,5,6-tetrahydro-1H-pyrano[4,3-b]thieno[3,2-d]pyridin-8-yl)-1H-pyrazole-1-carboxylate (50 mg, 0.13 mmol, 1.0 equiv.) in anhydrous THF (4.0 mL) was added t-BuMgCl (1 M in THF, 3.9 mL, 30 equiv.) at 0° C. The mixture was stirred at 0° C. for 0.5 h. The mixture was quenched with saturated aqueous NH4Cl (10 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography (SiO, 0-10% MeOH in DCM) to give racemic 4-(tert-butyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-3,4-dihydro-1H-pyrano[4,3-b]thieno[3,2-d]pyridin-6(5H)-one (10 mg, 23%). MS obsd. (ESI + ):346.5[(M+H) + ].

[0308] The individual enantiomers were separated via chiral SFC. (S)-4-(tert-butyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-3,4-dihydro-1H-pyrano[4,3-b]thieno[3,2-d]pyridin-6(5H)-one (compound 20): MS obsd. (ESI + ):346.1[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d6) δ ppm: 13.23 (s, 1H), 10.02 (s, 1H), 8.29 (s, 1H), 7.97 (s, 1H), 7.40 (s, 1H), 5.32 (s, 1H), 4.80 (m, 2H), 4.14 (d, J = 11.6 Hz, 1H), 3.46 (d, J = 11.6 Hz, 1H), 1.03 (s, 9H). (R)-4-(tert-butyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-3,4-dihydro-1H-pyrano[4,3-b]thieno[3,2-d]pyridin-6(5H)-one (compound 21): MS obsd. (ESI + ):346.1[(M+H)+ ].

[0309] Examples 22 and 23—Compounds 22 and 23: (S)-4-(tert-butyl)-4-hydroxy-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 22) and (R)-4-(tert-butyl)-4-hydroxy-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 23) (Stereochemistry for Compounds 22 and 23 is arbitrarily assigned) [ka] Step A: 4-(benzyloxy)-8-(pyridin-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] To a solution of methyl 3-bromo-5-(4-pyridyl)thiophene-2-carboxylate (1.0 g, 3.4 mmol, 1.0 equiv.) and 3-benzyloxytetrahydropyran-4-one (1.4 g, 6.7 mmol, 2.0 equiv.) in toluene (50.0 mL), CsCO (2.2 g, 6.71 mmol, 2.0 equiv.), Pd(dba) (614 mg, 0.67 mmol, 0.2 equiv.), Xantphos (582 mg, 1.01 mmol, 0.3 equiv.), and NaSO (64 mg, 0.34 mmol, 0.1 equiv.) were added at room temperature under a N atmosphere. The mixture was stirred at 105 °C for 16 h, cooled, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (SiO, 0-10% MeOH in DCM) to give 4-(benzyloxy)-8-(pyridin-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (457 mg, 32%). MS obsd. (ESI + ):392.1[(M+H) +].

[0310] Step B: 4-(benzyloxy)-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one: [ka] To a solution of 4-(benzyloxy)-8-(pyridin-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (1.0 g, 2.55 mmol, 1.0 equiv.) in MeOH (9.0 mL) was added NH4OH (9.0 mL). The mixture was heated in a microwave reactor at 100 °C for 3 h and monitored by LCMS until completion. The mixture was filtered, and the filter cake was washed with MeOH (50.0 mL) and dried under vacuum to give 4-(benzyloxy)-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (512 mg, 50%). MS obsd. (ESI + ):391.2[(M+H) + ].

[0311] Step C: 4-Hydroxy-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one: [ka] To a solution of 4-(benzyloxy)-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (475 mg, 1.22 mmol, 1.0 equiv.) in DCM (12.0 mL) was added BCl (1 M in DCM, 12.2 mL, 10.0 equiv.). The mixture was stirred at room temperature for 1 hour, then quenched with aqueous NaHCO (24.0 mL) and diluted with EtOAc (25.0 mL). The mixture was filtered, and the filter cake was washed with EtOAc (25.0 mL) and dried under vacuum to give 4-hydroxy-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (360 mg, 92%). MS obsd.(ESI + ):301.0[(M+H) + ].

[0312] Step D: 8-(pyridin-4-yl)-1,5-dihydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridine-4,6(3H)-dione: [ka] To a solution of 4-hydroxy-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (10 mg, 0.033 mmol, 1.0 equiv.) in DCM (1.0 mL) and DMF (1.0 mL) was added Dess-Martin periodinane (140 mg, 0.333 mmol, 10 equiv.). The mixture was stirred at room temperature for 2 hours, then quenched with saturated aqueous NaHCO (2.0 mL) and extracted with EtOAc (10 mL × 2). The combined organic phase was dried over sodium sulfate, filtered, and concentrated. The residue was purified by silica gel chromatography (SiO, 1-30% MeOH in DCM) to give 8-(pyridin-4-yl)-1,5-dihydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridine-4,6(3H)-dione (6 mg, 60%). MS obsd. (ESI + ):299.3[(M+H) +].

[0313] Step E: (S)-4-(tert-butyl)-4-hydroxy-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 22) and (R)-4-(tert-butyl)-4-hydroxy-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 23): [ka] To a solution of 8-(pyridin-4-yl)-1,5-dihydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridine-4,6(3H)-dione (25 mg, 0.08 mmol, 1.0 equiv.) in anhydrous THF (1.5 mL) was added t-BuMgBr (1 M in THF, 1.7 mL, 20.0 equiv.) at 0° C. The mixture was stirred at 0° C. for 2 hours. The mixture was quenched with saturated aqueous NH4Cl (2.0 mL) and extracted with EtOAc (20.0 mL × 3). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography (SiO, 0-10% MeOH in DCM) to give racemic 4-(tert-butyl)-4-hydroxy-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (5 mg, 17%). MS obsd. (ESI + ):357.1[(M+H) + ].

[0314] The individual enantiomers were separated via chiral SFC. (S)-4-(tert-butyl)-4-hydroxy-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 22): MS obsd. (ESI + ):357.2[(M+H) + ]; 1H NMR (400 MHz, DMSO-d6) δ ppm: 10.35 (s, 1H), 8.70 (d, J = 6.0 Hz, 2H), 8.03 (s, 1H), 7.82 (d, J = 6.0 Hz, 2H), 5.35 (s, 1H), 4.87 (m, 2H), 4.15 (d, J = 11.2 Hz, 1H), 3.47 (d, J = 11.2 Hz, 1H), 1.03 (s, 9H). (R)-4-(tert-butyl)-4-hydroxy-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 23): MS obsd. (ESI + ):357.2[(M+H) + ].

[0315] Examples 24 and 25—Compounds 24 and 25: (S)-4-hydroxy-4-isopropyl-8-(3-methyl-1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 24) and (R)-4-hydroxy-4-isopropyl-8-(3-methyl-1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 25) (Stereochemistry for Compounds 24 and 25 is arbitrarily assigned) [ka] Synthesized via a similar route to compounds 16 and 17. (S)-4-Hydroxy-4-isopropyl-8-(3-methyl-1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 24). MS obsd. (ESI + ):346.2[(M+H) + ]; 1H NMR (400 MHz, DMSO-d6) δ ppm: 12.94 (brs, 1H), 10.81 (brs, 1H), 7.96 (brs, 1H), 7.29 (s, 1H), 5.27 (s, 1H), 4.69 (s, 2H), 3.96 (d, J = 11.8 Hz, 1H), 3.53 (d, J = 11.6 Hz, 1H), 2.48 (s, 3H), 2.40-2.34 (m, 1H), 1.00 (d, J = 6.8 Hz, 3H), 0.76 (d, J = 6.8 Hz, 3H). (R)-4-Hydroxy-4-isopropyl-8-(3-methyl-1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 25). MS obsd.(ESI + ):346.2[(M+H) + ].

[0316] Examples 26 and 27 - Compounds 26 and 27: (R)-8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-4-isopropyl-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 26) and (S)-8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-4-isopropyl-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 27) (Stereochemistry for Compounds 26 and 27 is arbitrarily assigned) [ka] Step A: 4-bromo-3-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole: [ka] To a solution of 4-bromo-3-fluoro-1H-pyrazole (1.5 g, 9.1 mmol, 1.0 equiv) in DMF (15 mL) was added sodium hydride (60% in mineral oil, 700 mg, 18 mmol, 2.0 equiv) at 0 °C. The mixture was stirred at 0 °C for 10 minutes, and then 2-(chloromethoxy)ethyl-trimethyl-silane (2.27 g, 13.6 mmol, 2.41 mL, 1.5 equiv) was added. The mixture was stirred at room temperature for 2 hours and then quenched by adding saturated NH Cl (aq) followed by water (200 mL). The mixture was extracted with EtOAc (50 mL × 3), and the combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue, which was purified by flash column chromatography (SiO, 0–4% EtOAc in PE) to give 4-bromo-3-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (2.6 g, 96%). 1 H NMR(400MHz,CDCl3)δ ppm:7.45(d,J=2.0Hz,1H),5.24(d,J=0.8Hz,2H),3.58(m,2H),0.92(m,2H),0.00(s,9H).

[0317] Step B: 3-Fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole: [ka] A mixture of 4-bromo-3-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (850 mg, 2.88 mmol, 1.0 equiv), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (804 mg, 3.17 mmol, 1.1 equiv), Pd(dppf)Cl (421 mg, 576 μmol, 0.2 equiv), and potassium acetate (848 mg, 8.64 mmol, 3.0 equiv) in 1,4-dioxane (17.0 mL) was heated at 100° C. under nitrogen for 16 hours. The mixture was cooled and concentrated in vacuo, then water (50 mL) was added and the mixture was extracted with EtOAc (40 mL×3). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a residue which was purified by flash column chromatography (SiO, 0-8% EtOAc in PE) to give 3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (650 mg, 66%). 1 H NMR(400MHz,CDCl3)δ ppm:7.66(d,J=2.4Hz,1H),5.26(s,2H),3.59(m,2H),1.34(s,12H),0.92(m,2H),0.00(s,9H).

[0318] Step C: Methyl 3-bromo-5-(3-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)thiophene-2-carboxylate: [ka] A solution of methyl 3,5-dibromothiophene-2-carboxylate (1.37 g, 4.56 mmol, 1.2 equiv.), 3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (1.3 g, 3.80 mmol, 1.0 equiv.), Xantphos (440 mg, 760 μmol, 0.20 equiv.), KPO (2.4 g, 11 mmol, 3.0 equiv.), and Pd(OAc) (85 mg, 380 μmol, 0.10 equiv.) in THF (60.0 mL) was degassed with N and the mixture was stirred at 60 °C for 16 h. The mixture was filtered, concentrated in vacuo, and the residue was purified by flash column chromatography (SiO, 0-7% EtOAc in PE) to give 3-bromo-5-(3-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)thiophene-2-carboxylate (810 mg, 49%). MS obsd. (ESI + ): 79 Br / 81 Br 435.2, 437.2 [(M+H) + ].

[0319] Step D: 4-(benzyloxy)-8-(3-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-4-isopropyl-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] To a solution of 3-bromo-5-(3-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)thiophene-2-carboxylate (400 mg, 918.73 μmol, 1.0 equiv.), 3-(benzyloxy)-3-isopropyltetrahydro-4H-pyran-4-one (462 mg, 1.86 mmol, 2.0 equiv., prepared in a manner similar to Steps B–C of Compound 28), and cesium carbonate (909 mg, 2.79 mmol, 3.0 equiv.) in toluene (20.0 mL) was added Pd(dba) (170 mg, 186 μmol, 0.20 equiv.) and Xantphos (215 mg, 372 μmol, 0.40 equiv.). The mixture was degassed twice with N and stirred at 105 °C for 16 h. The mixture was filtered and concentrated in vacuo to give a residue that was purified by flash column chromatography (SiO, 0–16% EtOAc in PE) to give 4-(benzyloxy)-8-(3-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-4-isopropyl-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (275 mg, 51%). MS obsd. (ESI + ):571.3[(M+H) + ].

[0320] Step E: 8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-4-isopropyl-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] To a solution of 4-(benzyloxy)-8-(3-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-4-isopropyl-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (275 mg, 482 μmol, 1.0 equiv.) in DCM (12.0 mL) was added BCl (1 M, 1.93 mL, 4.0 equiv.) at 0° C., and the mixture was stirred at 0° C. for 40 min. The mixture was concentrated via a stream of nitrogen, and the pH was adjusted to approximately 9 by the slow addition of NHOH (aq.) at 0° C. The mixture was concentrated in vacuo to give 8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-4-isopropyl-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (400 mg, crude), which was used without further purification. MS obsd. (ESI + ):351.1[(M+H) + ].

[0321] Step F: (R)-8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-4-isopropyl-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 26) and (S)-8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-4-isopropyl-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 27): [ka] To a solution of 8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-4-isopropyl-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (400 mg, crude) in isopropanol (6.5 mL) was added aqueous NH4OH (6.5 mL). The mixture was stirred at 95 °C under microwave heating for 4 h, then concentrated and purified by flash column chromatography (SiO2, 0-6% MeOH in DCM) to give 8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-4-isopropyl-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (75 mg, 31% over two steps). MS obsd. (ESI + ):350.1[(M+H) + ].

[0322] The racemic mixture was separated via chiral SFC to give the individual enantiomers. (R)-8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-4-isopropyl-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 26): MS obsd. (ESI + ):350.0[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d6) δ ppm: 12.89 (brs, 1H), 10.90 (brs, 1H), 8.32 (d, J = 1.2 Hz, 1H), 7.35 (s, 1H), 5.27 (s, 1H), 4.68 (s, 2H), 3.96 (d, J = 12.0 Hz, 1H), 3.53 (d, J = 12.0 Hz, 1H), 2.41-2.34 (m, 1H), 1.00 (d, J = 6.8 Hz, 3H), 0.76 (d, J = 6.8 Hz, 3H). (S)-8-(3-Fluoro-1H-pyrazol-4-yl)-4-hydroxy-4-isopropyl-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 27): MS obsd. (ESI + ):350.0[(M+H) + ].

[0323] Examples 28 and 29—Compounds 28 and 29: (S)-4-hydroxy-8-(1H-pyrazol-4-yl)-4-(trifluoromethyl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 28) and (R)-4-hydroxy-8-(1H-pyrazol-4-yl)-4-(trifluoromethyl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 29) (Stereochemistry for Compounds 28 and 29 is arbitrarily assigned) [ka] Step A: 4,4-Dimethoxy-3-(trifluoromethyl)tetrahydro-2H-pyran-3-ol: [ka] To a solution of 4,4-dimethoxydihydro-2H-pyran-3(4H)-one (500 mg, 3.12 mmol, 1.0 equiv., synthesized according to the procedure described in WO2013 / 152269) in THF (5.0 mL) was added TBAF (81.6 mg, 312 μmol, 90.4 μL, 0.10 equiv.) at 0 °C. After the mixture was stirred at 0 °C for 10 min, trimethyl(trifluoromethyl)silane (666 mg, 4.68 mmol, 744 230 μL, 1.5 equiv.) was added dropwise. The mixture was allowed to warm to room temperature, where it was stirred for 16 h. The mixture was concentrated and purified by flash column chromatography (SiO2, 12–25% EtOAc in PE) to give 4,4-dimethoxy-3-(trifluoromethyl)tetrahydro-2H-pyran-3-ol (480 mg, 66%). 1 H NMR(400MHz,CDCl3)δ ppm:4.02-3.99(m,2H),3.85-3.82(m,2H),3.37(s,6H),2.03-2.02(m,2H).

[0324] Step B: 3-(benzyloxy)-4,4-dimethoxy-3-(trifluoromethyl)tetrahydro-2H-pyran: [ka] To a solution of 4,4-dimethoxy-3-(trifluoromethyl)tetrahydropyran-3-ol (1 g, 4.34 mmol, 1.0 equiv) in DMF (8.0 mL) was added NaH (333 mg, 8.69 mmol, 60% in mineral oil, 2.0 equiv) at 0 °C. The mixture was stirred at 0 °C for 10 min, and then bromomethylbenzene (1.49 g, 8.69 mmol, 1.03 mL, 2.0 equiv) was added dropwise. The mixture was stirred at room temperature for 3 h, quenched with saturated NH4Cl (aq), diluted with water (120 mL), and extracted with EtOAc (50 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated to give a residue which was purified by flash column chromatography (SiO, 0-7% EtOAc in PE) to give 3-(benzyloxy)-4,4-dimethoxy-3-(trifluoromethyl)tetrahydro-2H-pyran (690 mg, 49%). 1 H NMR(400MHz,CDCl3)δ ppm:7.38-7.20(m,5H),4.87(d,J=10.5Hz,1H),4.75(d,J=10.5Hz,1H),4.13-4.06(m,2 H),3.77-3.70(m,2H),3.39(s,3H),3.37(s,3H),2.22-2.16(m,1H),1.99-1.93(m,1H).

[0325] Step C: 3-(benzyloxy)-3-(trifluoromethyl)tetrahydro-4H-pyran-4-one: [ka] To a solution of 3-(benzyloxy)-4,4-dimethoxy-3-(trifluoromethyl)tetrahydro-2H-pyran (690 mg, 2.15 mmol, 1.0 equiv.) in acetone (15.0 mL) was added iodine (54.68 mg, 215.42 μmol, 0.1 equiv.). The reaction mixture was stirred at room temperature for 1 h, then quenched with saturated NaSO (aq.) (25 mL) and extracted into DCM (35 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (SiO, 0–8% EtOAc in PE) to give 3-(benzyloxy)-3-(trifluoromethyl)tetrahydro-4H-pyran-4-one (550 mg, 93%). 1 H NMR(400MHz,CDCl3)δ ppm:7.39-7.20(m,5H),4.90(d,J=11.0Hz,1H),4.62(d,J=11.0Hz,1H),4.33(d,J=12.8Hz,1H),4. 23-4.17(m,1H),3.90(d,J=12.8Hz,1H),3.88-3.81(m,1H),3.02-2.94(m,1H),2.54-2.48(m,1H).

[0326] Step D: 4-(benzyloxy)-4-(trifluoromethyl)-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] To a solution of methyl 3-bromo-5-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)thiophene-2-carboxylate (200 mg, 480 μmol, 1.0 equiv.), 3-benzyloxy-3-(trifluoromethyl)tetrahydropyran-4-one (263 mg, 958 μmol, 2.0 equiv.), and cesium carbonate (468 mg, 1.44 mmol, 3.0 equiv.) in toluene (12.0 mL) was added Pd(dba) (87.8 mg, 95.8 μmol, 0.2 equiv.) and Xantphos (111 mg, 192 μmol, 0.4 equiv.). The mixture was degassed with N and stirred at 105 °C for 16 h. The mixture was filtered, concentrated in vacuo, and then purified by flash column chromatography (SiO, 0-20% EtOAc in PE) to give 4-(benzyloxy)-4-(trifluoromethyl)-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (185 mg, 66%). MS obsd. (ESI + ):579.5[(M+H) + ].

[0327] Step E: 4-(benzyloxy)-4-(trifluoromethyl)-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one: [ka] To a solution of 4-(benzyloxy)-4-(trifluoromethyl)-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (185 mg, 320 μmol) in MeOH (5.0 mL) was added ammonium hydroxide (5.0 mL). The mixture was stirred at 100 °C in a microwave reactor for 6 h and then concentrated in vacuo to give a residue that was purified by flash column chromatography (SiO, 0-4% MeOH in DCM) to give 4-(benzyloxy)-4-(trifluoromethyl)-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (105 mg, 57%). MS obsd. (ESI + ):578.3[(M+H) + ].

[0328] Step F: (S)-4-hydroxy-8-(1H-pyrazol-4-yl)-4-(trifluoromethyl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 28) and (R)-4-hydroxy-8-(1H-pyrazol-4-yl)-4-(trifluoromethyl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 29): [ka] 4-(Benzyloxy)-4-(trifluoromethyl)-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (105 mg, 181.76 μmol, 1.0 equiv) was dissolved in trifluoroacetic acid (2.6 mL) at 0° C. The mixture was stirred at 60° C. under microwave irradiation for 2 h, then concentrated, and the pH was adjusted to approximately 9 by slow addition of NH / MeOH (7 M). The mixture was concentrated in vacuo and purified by flash column chromatography (SiO, 0-6% MeOH in DCM) to give racemic 4-hydroxy-8-(1H-pyrazol-4-yl)-4-(trifluoromethyl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (54 mg, 83%). MS obsd. (ESI + ):358.0[(M+H) + ].

[0329] The individual enantiomers were separated via chiral SFC. (S)-4-Hydroxy-8-(1H-pyrazol-4-yl)-4-(trifluoromethyl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 28): MS obsd. (ESI + ):358.0[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d6) δ ppm: 13.25 (s, 1H), 10.94 (s, 1H), 8.27 (brs, 2H), 7.48 (s, 1H), 7.13 (s, 1H), 4.89-4.77 (m, 2H), 4.23 (d, J = 12.4 Hz, 1H), 3.71 (m, 1H). (R)-4-Hydroxy-8-(1H-pyrazol-4-yl)-4-(trifluoromethyl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 29): MS obsd. (ESI + ):358.0[(M+H) + ].

[0330] Examples 30 and 31 - Compounds 30 and 31: (S)-4-hydroxy-8-(1-methyl-1H-pyrazol-5-yl)-4-(trifluoromethyl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 30) and (R)-4-hydroxy-8-(1-methyl-1H-pyrazol-5-yl)-4-(trifluoromethyl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 31) (Stereochemistry for Compounds 30 and 31 is arbitrarily assigned) [ka] Synthesized by a similar route to compounds 28 and 29. (S)-4-Hydroxy-8-(1-methyl-1H-pyrazol-5-yl)-4-(trifluoromethyl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 30). MS obsd. (ESI + ):372.1[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d6) δ ppm: 11.21 (brs, 1H), 7.65 (s, 1H), 7.54 (d, J = 2.0 Hz, 1H), 7.19 (brs, 1H), 6.72 (d, J = 2.0 Hz, 1H), 4.96 (d, J = 15.2 Hz, 1H), 4.81 (d, J = 15.2 Hz, 1H), 4.25 (d, J = 12.0 Hz, 1H), 4.05 (s, 3H), 3.71 (d, J = 12.0 Hz, 1H). (R)-4-Hydroxy-8-(1-methyl-1H-pyrazol-5-yl)-4-(trifluoromethyl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 31). MS obsd.(ESI + ):372.1[(M+H) + ].

[0331] Examples 32 and 33 - Compounds 32 and 33: (S)-4-hydroxy-8-(5-methyl-1H-pyrazol-4-yl)-4-(trifluoromethyl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 32) and (R)-4-hydroxy-8-(5-methyl-1H-pyrazol-4-yl)-4-(trifluoromethyl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 33) (stereochemistry for Compounds 32 and 33 is arbitrarily assigned). [ka] Synthesized by a similar route to compounds 28 and 29. (S)-4-Hydroxy-8-(5-methyl-1H-pyrazol-4-yl)-4-(trifluoromethyl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one. MS obsd. (ESI + ):372.0[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d6) δ ppm: 12.98 (brs, 1H), 10.90 (brs, 1H), 8.23-7.86 (m, 1H), 7.35 (s, 1H), 7.14 (s, 1H), 4.92 (d, 15.02 Hz, 1H), 4.78 (d, 15.02 Hz, 1H), 4.24 (d, J = 12.0 Hz, 1H), 3.70 (d, J = 12.0 Hz, 1H), 2.43 (s, 3H). (R)-4-Hydroxy-8-(5-methyl-1H-pyrazol-4-yl)-4-(trifluoromethyl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one. MS obsd. (ESI + ):372.0[(M+H) + ].

[0332] Examples 34 and 35—Compounds 34 and 35: (S)-4-(hydroxymethyl)-4-methyl-8-(1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (Compound 34) and (R)-4-(hydroxymethyl)-4-methyl-8-(1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (Compound 35) (stereochemistry for Compounds 34 and 35 is arbitrarily assigned) [ka] Step A: Benzyl 2-(2-hydroxytetrahydrofuran-2-yl)acetate: [ka] A mixture of benzyl 2-bromoacetate (19.2 g, 83.6 mmol, 1.2 equiv.), indium (9.60 g, 83.6 mmol, 1.2 equiv.), tetrahydrofuran-2-one (6.0 g, 69.7 mmol, 1.0 equiv.), and THF (21.0 mL) was stirred at 70 °C for 16 h. The reaction was quenched with saturated aqueous NaHCO (20 mL) and extracted with EtOAc (60 mL × 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by flash column chromatography (SiO, 10–20% EtOAc in DCM) gave benzyl 2-(2-hydroxytetrahydrofuran-2-yl)acetate (4.0 g, 24%). MS obsd. (ESI + ):219.2[(M+H-H2O) + ].

[0333] Step B: Benzyl 2-diazo-6-hydroxy-3-oxohexanoate: [ka] To a solution of benzyl 2-(2-hydroxytetrahydrofuran-2-yl)acetate (700 mg, 2.96 mmol, 1.0 equiv.) and N-diazo-2,4,6-triisopropyl-benzenesulfonamide (1.01 g, 3.26 mmol, 1.1 equiv.) in THF (10.0 mL) was added N,N-diethylethanamine (900 mg, 8.9 mmol, 3.0 equiv.) at 0 °C. The mixture was stirred at room temperature for 16 h and then concentrated. The residue was purified by flash column chromatography (SiO2, 15-30% EtOAc in PE) to give 2-diazo-6-hydroxy-3-oxo-hexanoate (1.18 g, 81%). MS obsd. (ESI + ):263.2[(M+H) + ].

[0334] Step C: Benzyl 3-oxotetrahydro-2H-pyran-2-carboxylate: [ka] A solution of benzyl 2-diazo-6-hydroxy-3-oxo-hexanoate (1.0 g, 3.81 mmol, 1.0 equiv.) in benzene (50.0 mL) was added dropwise over 35 min to a solution of Rh(OAc) (50.0 mg, 113 μmol, 0.030 equiv.) in benzene (50.0 mL) at 100° C. The mixture was stirred at 100° C. for 1 h, then filtered and concentrated to give benzyl 3-oxotetrahydro-2H-pyran-2-carboxylate (180 mg, crude), which was used in the next step without purification. MS obsd. (ESI + ):235.2[(M+H) + ].

[0335] Step D: Benzyl 2-methyl-3-oxotetrahydro-2H-pyran-2-carboxylate: [ka] To a solution of benzyl 3-oxotetrahydro-2H-pyran-2-carboxylate (890 mg, 3.80 mmol, 1.0 equiv) in DMF (15 mL) was added sodium hydride (247 mg, 5.70 mmol, 60% in mineral oil, 1.5 equiv) at -20 °C. The mixture was stirred for 10 min, then iodomethane (1.62 g, 11.40 mmol, 3.0 equiv) was added, and the mixture was stirred at room temperature for 16 h. The mixture was poured into water and extracted with ethyl acetate (30 mL × 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo, and the residue was purified by flash column chromatography (SiO2, 0–10% EtOAc in PE) to give benzyl 2-methyl-3-oxotetrahydro-2H-pyran-2-carboxylate (474 ​​mg, 50%). MS obsd. (ESI + ):271.0[(M+Na) + ].

[0336] Step E: Benzyl 4-methyl-6-oxo-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,2,4,6-tetrahydropyrano[3,4-b]thieno[3,2-d]pyran-4-carboxylate: [ka] Methyl 3-bromo-5-[1-(2-trimethylsilylethoxymethyl)pyrazol-4-yl]thiophene-2-carboxylate (84.1 mg, 201 μmol, 1.0 equiv.), benzyl 2-methyl-3-oxotetrahydro-2H-pyran-2-carboxylate (100 mg, 403 μmol, 2.0 equiv.), sodium metabisulfite (11.5 mg, 60.4 μmol, 0.3 equiv.), CsCO (197 mg, 604 μmol, 3.0 equiv.), Sphos-Pd-G (31.4 mg, 40.3 μmol, 0.2 equiv.), and toluene (15.0 mL) were stirred at 105 °C for 16 h. The mixture was poured into water and extracted with EtOAc (70 mL × 2). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (SiO, 0–25% EtOAc in PE) to give benzyl 4-methyl-6-oxo-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,2,4,6-tetrahydropyrano[3,4-b]thieno[3,2-d]pyran-4-carboxylate (78 mg, 70%). MS obsd. (ESI + ):553.5[(M+H) + ].

[0337] Step F: 4-(hydroxymethyl)-4-methyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,4-dihydropyrano[3,4-b]thieno[3,2-d]pyran-6(2H)-one: [ka] A mixture of benzyl 4-methyl-6-oxo-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,2,4,6-tetrahydropyrano[3,4-b]thieno[3,2-d]pyran-4-carboxylate (50 mg, 90.5 μmol, 1.0 equiv.), calcium dichloride (50.2 mg, 452 μmol, 5 equiv.), and EtOH (5.0 mL) was stirred at room temperature for 20 minutes, and then sodium borohydride (51.3 mg, 1.36 mmol, 15.0 equiv.) was added portionwise to the mixture at 0° C., followed by stirring at room temperature for 20 minutes. The mixture was poured into water and extracted with EtOAc (30 mL × 2). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (SiO, 50% EtOAc in PE) to give 4-(hydroxymethyl)-4-methyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,4-dihydropyrano[3,4-b]thieno[3,2-d]pyran-6(2H)-one (20 mg, 49%). MS obsd. (ESI + ):449.2[(M+H) + ].

[0338] Step G: 4-(hydroxymethyl)-4-methyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one: [ka] A mixture of 4-(hydroxymethyl)-4-methyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,4-dihydropyrano[3,4-b]thieno[3,2-d]pyran-6(2H)-one (60 mg, 134 μmol), ammonium hydroxide (3.5 mL), and MeOH (3.5 mL) was stirred in a microwave reactor at 100° C. for 3 hours. The mixture was concentrated in vacuo, and the residue was purified by flash column chromatography (SiO, 15–35% EtOAc in PE) to give 4-(hydroxymethyl)-4-methyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (20 mg, 33%). MS obsd. (ESI + ):448.5[(M+H) + ].

[0339] Step H: (S)-4-(hydroxymethyl)-4-methyl-8-(1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (compound 34) and (R)-4-(hydroxymethyl)-4-methyl-8-(1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (compound 35): [ka] Trichloroborane (1 M, 890 μL, 5.0 equiv.) was added to a solution of 4-(hydroxymethyl)-4-methyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (80 mg, 180 μmol) in DCM (50.0 mL) at 0° C. The mixture was stirred at room temperature for 2 h, then quenched with MeOH and concentrated in vacuo. The residue was basified with NaHCO and purified by column chromatography (SiO, 0-6% MeOH in DCM) to give racemic 4-(hydroxymethyl)-4-methyl-8-(1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (32 mg, 56%). MS obsd. (ESI + ):318.5[(M+H) + ].

[0340] The individual enantiomers were separated via chiral SFC. (S)-4-(hydroxymethyl)-4-methyl-8-(1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (compound 34): MS obsd. (ESI + ):318.0[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d6) δ ppm: 13.22 (brs, 1H), 10.90 (brs, 1H), 8.31 (s, 1H), 7.97 (s, 1H), 7.48 (s, 1H), 4.87 (s, 1H), 4.09-3.96 (m, 1H), 3.93-3.72 (m, 2H), 3.51 (d, J = 11.6 Hz, 1H), 2.86-2.58 (m, 2H), 1.39 (s, 3H). (R)-4-(hydroxymethyl)-4-methyl-8-(1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (compound 35). MS obsd. (ESI + ):318.0[(M+H) + ].

[0341] Examples 36 and 37 - Compounds 36 and 37: (S)-6-(difluoromethyl)-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 36) and (R)-6-(difluoromethyl)-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 37) (Stereochemistry for Compounds 36 and 37 is arbitrarily assigned) [ka] Step A: 2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-8,9-dihydro-4H-thieno[2,3-c]chromene-4,6(7H)-dione: [ka] Methyl 3-bromo-5-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)thiophene-2-carboxylate (200 mg, 480 μmol, 1.0 equiv.) and cyclohexane-1,2-dione (1.1 g, 9.6 mmol, 20 equiv.) were dissolved in anhydrous toluene (40.0 mL), followed by the addition of Sphos-Pd-G3 (56 mg, 96 μmol, 0.20 equiv.), Cs2CO3 (470 mg, 1.40 mmol, 3.0 equiv.), and Na2SO5 (18 mg, 0.096 mmol, 0.20 equiv.). The mixture was degassed with N2 and heated to 105 °C for 16 h. The mixture was cooled, concentrated in vacuo, and purified by flash column chromatography (SiO2, 0–45% EtOAc in PE). Further purification (SiO, 0-8% MeOH in DCM) gave 2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-8,9-dihydro-4H-thieno[2,3-c]chromene-4,6(7H)-dione (20 mg, 10%). MS obsd. (ESI + ):417.5[(M+H) +].

[0342] Step B: 6-(difluoromethyl)-6-hydroxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one: [ka] Cesium fluoride (55 mg, 0.36 mmol, 2.5 equiv.) was added to a solution of 2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-8,9-dihydro-4H-thieno[2,3-c]chromene-4,6(7H)-dione (60 mg, 0.144 mmol, 1.0 equiv.) in anhydrous DMF (4.0 mL) under a N atmosphere, followed by the addition of difluoromethyl(trimethyl)silane (90 mg, 0.72 mmol, 100 μL, 5.0 equiv.). The mixture was stirred at room temperature for 24 h, then diluted with water (50 mL) and extracted with EtOAc (40 mL × 4). The combined organic phase was dried (NaSO), filtered, and concentrated. Purification (SiO2, 0-39% EtOAc in PE) gave 6-(difluoromethyl)-6-hydroxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (23 mg, 34%). MS obsd. (ESI+): 469.4 [(M+H) + ].

[0343] Step C: 6-(difluoromethyl)-6-hydroxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one: [ka] 6-(Difluoromethyl)-6-hydroxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (65 mg, 0.14 mmol) was added to NHOH (5.0 mL, saturated aqueous solution), followed by MeOH (5.0 mL). The mixture was heated to 95° C. in a microwave reactor for 10 hours and then concentrated to give 6-(difluoromethyl)-6-hydroxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (65 mg, crude). This material was used in the next step without further purification. MS obsd.(ESI+):468.5[(M+H) + ].

[0344] Step D: (S)-6-(difluoromethyl)-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 36) and (R)-6-(difluoromethyl)-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 37): [ka] 6-(Difluoromethyl)-6-hydroxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (180 mg, 370 μmol) was dissolved in DCM (15.0 mL) and cooled to 0 °C. Trifluoroacetic acid (3.0 mL) was added. The reaction mixture was warmed to room temperature, and after 4 h, the mixture was concentrated in vacuo. The residue was purified (0–25% MeCN (0.1% NHOH) in C18 SiO2 water) to give racemic 6-(difluoromethyl)-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (40 mg, 26%). MS obsd.(ESI + ):388.4[(M+H) + ].

[0345] The individual enantiomers were separated via chiral SFC. (S)-6-(Difluoromethyl)-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 36): MS obsd. (ESI + ):388.1[(M+H) + ].(R)-6-(Difluoromethyl)-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 37): MS obsd. (ESI+): 388.1 [(M+H) + ]; 1 H NMR(400MHz,DMSO-d6)δ ppm:13.22(s,1H),10.68(s,1H),8.32(s,1H),7.98(s,1H),7.54(s,1H),6.52(t,J=55.6Hz,1 H),6.26(s,1H),2.77-2.73(m,1H),2.68-2.61(m,1H),2.10-2.04(m,1H),1.87-1.85(m,3H).

[0346] Examples 38 and 39—Compounds 38 and 39: (S)-9-methoxy-9-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 38) and (R)-9-methoxy-9-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 39) (Stereochemistry for Compounds 38 and 39 is arbitrarily assigned) [ka] Step A: 7,8-Dihydro-4H-thieno[2,3-c]chromene-4,9(6H)-dione: [ka] Methyl 3-bromothiophene-2-carboxylate (0.50 g, 2.3 mmol, 1.0 equiv.) and cyclohexane-1,3-dione (1.5 g, 14 mmol, 6.0 equiv.) were dissolved in toluene (80.0 mL), followed by the addition of CsCO (2.2 g, 6.8 mmol, 3.0 equiv.) and SPhos-Pd-G (180 mg, 230 μmol, 0.10 equiv.). The mixture was heated to 105 °C for 16 h, cooled, and diluted with HO (50 mL). The mixture was extracted with EtOAc (50 mL × 3), and the combined organic phase was washed with brine (50 mL), dried (NaSO), filtered, and concentrated. Purification by column chromatography (SiO, 0-20% EtOAc in PE) gave 7,8-dihydro-4H-thieno[2,3-c]chromene-4,9(6H)-dione (82 mg, 14%). MS obsd. (ESI + ):221.2[(M+H) + ].

[0347] Step B: 7,8-Dihydrothieno[2,3-c]quinoline-4,9(5H,6H)-dione: [ka] 7,8-Dihydro-4H-thieno[2,3-c]chromene-4,9(6H)-dione (550 mg, 2.50 mmol, 1.0 equiv) was dissolved in MeOH (8.0 mL) followed by the addition of NH4OH (8 mL, saturated aqueous solution). The reaction mixture was heated to 90 °C in a microwave reactor for 4 h, cooled, and concentrated in vacuo. Purification by column chromatography (SiO2, 0-25% MeOH in DCM) gave 7,8-dihydrothieno[2,3-c]quinoline-4,9(5H,6H)-dione (420 mg, 77%). MS obsd. (ESI + ):220.2[(M+H) + ].

[0348] Step C: 5-((2-(trimethylsilyl)ethoxy)methyl)-7,8-dihydrothieno[2,3-c]quinoline-4,9(5H,6H)-dione: [ka] A solution of 7,8-dihydrothieno[2,3-c]quinoline-4,9(5H,6H)-dione (1.3 g, 5.9 mol, 1.0 equiv.) in anhydrous DMF (60.0 mL) was cooled to 0 °C, followed by the addition of NaH (360 mg, 8.9 mmol, 60% in mineral oil, 1.5 equiv.). After 30 min, 2-(chloromethoxy)ethyl-trimethyl-silane (2.0 g, 12 mmol, 2.1 mL, 2.0 equiv.) was added, and the mixture was warmed to room temperature, where it was stirred for 2 h. Water (150 mL) was added, and the mixture was extracted with EtOAc (100 mL × 3). The combined organic phases were washed (brine, 100 mL), dried (NaSO), filtered, and concentrated in vacuo. Purification by column chromatography (SiO, 0-30% EtOAc in PE) gave 5-((2-(trimethylsilyl)ethoxy)methyl)-7,8-dihydrothieno[2,3-c]quinoline-4,9(5H,6H)-dione (1.0 g, 48%). MS obsd. (ESI + ):350.5[(M+H) + ].

[0349] Step D: 9-Hydroxy-9-methyl-5-((2-(trimethylsilyl)ethoxy)methyl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one: [ka] 5-((2-(trimethylsilyl)ethoxy)methyl)-7,8-dihydrothieno[2,3-c]quinoline-4,9(5H,6H)-dione (1.0 g, 2.9 mmol, 1.0 equiv.) was dissolved in THF (30.0 mL). The solution was cooled to 0 °C, and then methylmagnesium bromide (1 M, 57.0 mL, 57 mmol, 20 equiv.) was added. The reaction mixture was warmed to room temperature and stirred for 2 h, after which water (100 mL) was added. The mixture was extracted with EtOAc (100 mL × 3), and the combined organic phases were washed with brine (100 mL), dried (NaSO), filtered, and concentrated in vacuo. Purification by column chromatography (0-30% EtOAc on SiO2PE) gave 9-hydroxy-9-methyl-5-((2-(trimethylsilyl)ethoxy)methyl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (320 mg, 27%). MS obsd. (ESI + ):366.5[(M+H) + ].

[0350] Step E: 9-Methoxy-9-methyl-5-((2-(trimethylsilyl)ethoxy)methyl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one: [ka] Sodium hydride (145 mg, 3.61 mmol, 60% in mineral oil, 4.0 equiv.) and iodomethane (1.3 g, 9.0 mmol, 0.6 mL, 10.0 equiv.) were added to a solution of 9-hydroxy-9-methyl-5-((2-(trimethylsilyl)ethoxy)methyl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (330 mg, 0.90 mmol, 1.0 equiv.) in anhydrous THF (15.0 mL). After 2 h, water (50 mL) was added, and the mixture was extracted with EtOAc (30 mL × 3). The combined organic phases were washed with brine (30 mL), dried (NaSO), filtered, and concentrated in vacuo. Purification by column chromatography (SiO, 0-15% EtOAc in PE) gave 9-methoxy-9-methyl-5-((2-(trimethylsilyl)ethoxy)methyl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (190 mg, 55%). MS obsd. (ESI + ):380.6[(M+H) + ].

[0351] Step F: 2-iodo-9-methoxy-9-methyl-5-((2-(trimethylsilyl)ethoxy)methyl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one: [ka] 9-Methoxy-9-methyl-5-((2-(trimethylsilyl)ethoxy)methyl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (140 mg, 0.37 mmol, 1.0 equiv) was dissolved in THF (8.0 mL) and cooled to −65° C. Lithium diisopropylamine (2 M, 0.7 mL, 1.5 mmol, 4.0 equiv) was added, followed by iodine (190 mg, 0.74 mmol, 2.0 equiv), and the mixture was stirred at −65° C. for 2 h. Water (10 mL) was added, and the mixture was extracted with EtOAc (10 mL × 3). The combined organic phases were washed with brine (10 mL), dried (NaSO), filtered, and concentrated in vacuo. Purification by column chromatography (SiO, 0-15% EtOAc in PE) gave 2-iodo-9-methoxy-9-methyl-5-((2-(trimethylsilyl)ethoxy)methyl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (120 mg, 62%). MS obsd. (ESI + ):506.6[(M+H) + ].

[0352] Step G: 9-Methoxy-9-methyl-5-((2-(trimethylsilyl)ethoxy)methyl)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one: [ka] Pd(dppf)Cl2 (32 mg, 40 μmol, 0.20 equiv.), XPhos (42 mg, 90 μmol, 0.40 equiv.), and Na2CO3 (70 mg, 0.65 mmol, 3.0 equiv.) were dissolved in HO / 1,4-dioxane (8.0 mL, 1:3 mixture) to prepare 2-iodo-9-methoxy-9-methyl-5-((2-(trimethylsilyl)ethoxy)methyl)- To a solution of 6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (110 mg, 0.22 mmol, 1.0 equiv.) and trimethyl-[2-[[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazol-1-yl]methoxy]ethyl]silane (110 mg, 0.35 mmol, 1.6 equiv.) was added. The mixture was heated to 110° C. for 2 h and then concentrated in vacuo. Purification by column chromatography (0-30% EtOAc in PE) gave 9-methoxy-9-methyl-5-((2-(trimethylsilyl)ethoxy)methyl)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (110 mg, 88%). MS obsd. (ESI + ):576.9[(M+H) + ].

[0353] Step H: (9S)-9-Methoxy-9-methyl-2-(1H-pyrazol-4-yl)-5,6,7,8-tetrahydrothieno[2,3-c]quinolin-4-one (compound 38) and (9R)-9-methoxy-9-methyl-2-(1H-pyrazol-4-yl)-5,6,7,8-tetrahydrothieno[2,3-c]quinolin-4-one (compound 39): [ka] BCl (1.0 M, 1.1 mL, 1.1 mmol, 5.0 equiv) was added to a solution of 9-methoxy-9-methyl-5-((2-(trimethylsilyl)ethoxy)methyl)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (130 mg, 0.22 mmol, 1.0 equiv) in DCM (13.0 mL) at 0° C. After 10 min, NH / MeOH was added (to pH 9) and the mixture was concentrated in vacuo. Purification by column chromatography (C18 SiO2, 0-20% MeCN in water (0.1% NH4OH)) gave racemic 9-methoxy-9-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (24 mg, 34%). MS obsd. (ESI + ):316.2[(M+H) + ].

[0354] The individual enantiomers were separated via chiral SFC. (9S)-9-Methoxy-9-methyl-2-(1H-pyrazol-4-yl)-5,6,7,8-tetrahydrothieno[2,3-c]quinolin-4-one (compound 38): MS obsd. (ESI + ):316.2[(M+H) + ]. (9R)-9-Methoxy-9-methyl-2-(1H-pyrazol-4-yl)-5,6,7,8-tetrahydrothieno[2,3-c]quinolin-4-one (compound 39): MS obsd. (ESI + ):316.2[(M+H) + ]; 1 H NMR(400MHz,DMSO-d6)δ ppm:13.21(s,1H),11.29(s,1H),8.32(s,1H),7.91(s,1H),7.64(s,1H),2.94(s,3H),2.70-2.52 (m,2H),2.15-2.01(m,1H),1.95-1.87(m,1H),1.83-1.67(m,1H),1.55-1.65(m,1H),1.50(m,3H).

[0355] Examples 40 and 41 - Compounds 40 and 41: (S)-6-hydroxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 40) and (R)-6-hydroxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 41) (Stereochemistry for Compounds 40 and 41 is arbitrarily assigned) [ka] Step A: tert-butyl 4-(6-hydroxy-4-oxo-4,5,6,7,8,9-hexahydrothieno[2,3-c]quinolin-2-yl)-1H-pyrazole-1-carboxylate: [ka] Triethylamine (280 mg, 2.8 mmol, 2.0 equiv.), DMAP (85 mg, 70 μmol, 0.50 equiv.), and (Boc)O (300 mg, 1.40 mmol, 1.0 equiv.) were added to a suspension of 6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-5H-thieno[2,3-c]quinolin-4-one (400 mg, 1.40 mmol, 1.0 equiv.) in DMF (20.0 mL). The mixture was stirred at room temperature for 16 hours and then extracted with EtOAc (50 mL × 3). The combined organic phase was washed with brine, dried (NaSO), filtered, and concentrated. Purification by column chromatography (SiO, 0-100% EtOAc in PE) gave tert-butyl 4-(6-hydroxy-4-oxo-4,5,6,7,8,9-hexahydrothieno[2,3-c]quinolin-2-yl)-1H-pyrazole-1-carboxylate (400 mg, 74%). MS obsd. (ESI + ):388.5[(M+H) + ].

[0356] Step B: tert-Butyl 4-(4,6-dioxo-5,7,8,9-tetrahydrothieno[2,3-c]quinolin-2-yl)pyrazole-1-carboxylate: [ka] A suspension of tert-butyl 4-(6-hydroxy-4-oxo-4,5,6,7,8,9-hexahydrothieno[2,3-c]quinolin-2-yl)-1H-pyrazole-1-carboxylate (140 mg, 0.36 mmol, 1.0 equiv.) and (1,1-diacetoxy-3-oxo-1,2-benziodoxol-1-yl)acetate (110 mg, 3.6 mmol, 10 equiv.) in DCM (14.0 mL) was stirred at room temperature for 1 h. The mixture was concentrated, and the residue was purified by column chromatography (SiO, 0–10% MeOH in DCM) to give tert-butyl 4-(4,6-dioxo-5,7,8,9-tetrahydrothieno[2,3-c]quinolin-2-yl)pyrazole-1-carboxylate (120 mg, 87%). MS obsd.(ESI + ):386.5[(M+H) + ].

[0357] Step C: (S)-6-hydroxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 40) and (R)-6-hydroxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 41): [ka] Methylmagnesium bromide (1.0 M, 4.8 mL, 4.8 mmol, 3.0 equiv) was added to a solution of tert-butyl 4-(6-hydroxy-4-oxo-4,5,6,7,8,9-hexahydrothieno[2,3-c]quinolin-2-yl)-1H-pyrazole-1-carboxylate (60 mg, 0.16 mmol, 1.0 equiv) in THF (24.0 mL). The mixture was stirred at room temperature for 2 h, then MeOH was added. The resulting mixture was concentrated in vacuo and purified by column chromatography (SiO, 0–10% MeOH in DCM) to give racemic 6-hydroxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (39 mg, 62%).

[0358] The individual enantiomers were separated via chiral SFC. (S)-6-hydroxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 40): MS obsd. (ESI + ):302.1[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d6) δ ppm: 13.10 (s, 1H), 10.51 (s, 1H), 8.12 (s, 2H), 7.46 (s, 1H), 5.12 (s, 1H), 2.32-2.51 (m, 2H), 1.88-1.82 (m, 3H), 1.78-1.67 (m, 1H), 1.46 (s, 3H). (R)-6-Hydroxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 41): MS obsd. (ESI + ):302.1[(M+H) + ].

[0359] Examples 42 and 43—Compounds 42 and 43: (S)-4-(difluoromethyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 42) and (R)-4-(difluoromethyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 43) (stereochemistry for Compounds 42 and 43 is arbitrarily assigned) [ka] Step A: 4-hydroxy-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] Sodium hydride (140 mg, 3.4 mmol, 60% in mineral oil, 2.0 equiv.) was added to a cooled (0 °C) solution of 4-hydroxy-8-(1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (500 mg, 1.7 mmol, 1.0 equiv.) in DMF (20.0 mL). After 10 min at 0 °C, SEMCl (370 μL, 350 mg, 2.0 mmol, 1.2 equiv.) was slowly added to the mixture. The mixture was stirred at room temperature for 2 h, then quenched with ammonium chloride (saturated aqueous solution), and then extracted with EtOAc (25 mL × 3). The combined organic phase was treated with brine, dried (Na2SO4), filtered, and concentrated. Purification by column chromatography (SiO, 0-40% EtOAc in DCM) gave 4-hydroxy-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (400 mg, 52%). MS obsd. (ESI+ ):421.2[(M+H) + ].

[0360] Step B: 8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1H,3H-pyrano[4,3-b]thieno[3,2-d]pyran-4,6-dione: [ka] 4-Hydroxy-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (400 mg, 950 μmol, 1.0 equiv.) was dissolved in DCM (20.0 mL) and DMF (20.0 mL), and (1,1-diacetoxy-3-oxo-1,2-benziodoxol-1-yl)acetate (1.2 g, 2.9 mmol, 3.0 equiv.) was added. The mixture was stirred at room temperature for 16 h and then quenched with NaSO (saturated aqueous solution) and NaHCO (saturated aqueous solution). The mixture was extracted with EtOAc (30 mL × 3), and the combined organic phase was dried (NaSO), filtered, and concentrated. Purification by column chromatography (SiO, 0-30% EtOAc in DCM) gave 8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1H,3H-pyrano[4,3-b]thieno[3,2-d]pyran-4,6-dione (270 mg, 63%). MS obsd. (ESI + ):419.2[(M+H) + ], 436.2[(M+NH4) + ].

[0361] Step C: 4-(difluoromethyl)-4-hydroxy-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] Cesium fluoride (210 mg, 1.4 mmol, 2.5 equiv) was added to a solution of 8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1H,3H-pyrano[4,3-b]thieno[3,2-d]pyran-4,6-dione (270 mg, 560 μmol, 1.0 equiv) in DMF (24.0 mL), followed by TMSCF2H (350 mg, 2.8 mmol, 400 μL, 5.0 equiv). The mixture was stirred for 16 h. Water was added, and the mixture was extracted with EtOAc (25 mL × 5). The combined organic phases were washed with brine, dried (Na2SO4), filtered, and concentrated. Purification by column chromatography (SiO, 0-40% EtOAc in PE) gave 4-(difluoromethyl)-4-hydroxy-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (48 mg, 16%). MS obsd. (ESI + ):471.4[(M+H) + ].

[0362] Step D: 4-(difluoromethyl)-4-hydroxy-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one: [ka] Aqueous ammonia (8.0 mL, 25% w / w) was added to a solution of 4-(difluoromethyl)-4-hydroxy-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (140 mg, 260 μmol, 1.0 equiv.) in i-PrOH (8.0 mL). The mixture was heated to 100° C. in a microwave reactor for 6 h, then cooled and concentrated. Purification by column chromatography (SiO, 0-8% MeOH in DCM) gave 4-(difluoromethyl)-4-hydroxy-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (55 mg, 41%). MS obsd. (ESI + ):470.2[(M+H) + ].

[0363] Step E: (S)-4-(difluoromethyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 42) and (R)-4-(difluoromethyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 43): [ka] 4-(Difluoromethyl)-4-hydroxy-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (10 mg, 21 μmol) was dissolved in DCM:trifluoroacetic acid (4.0 mL, 3:1). The mixture was stirred at room temperature for 1 h, then concentrated and purified by reverse-phase HPLC (C18 SiO2, 0-15% MeCN in water (0.1% NH4OH)). Further purification by column chromatography (SiO, 0-8% MeOH in DCM) gave racemic 4-(difluoromethyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (1.3 mg, 17%). MS obsd. (ESI + ):340.0[(M+H) + ].

[0364] The individual enantiomers were separated via chiral SFC. (S)-4-(Difluoromethyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 42): MS obsd. (ESI + ):340.2[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d6) δ ppm: 13.24 (s, 1H), 11.04 (s, 1H), 8.29 (s, 1H), 7.99 (s, 1H), 7.47 (s, 1H), 6.57 (s, 1H), 6.44 (t, J = 55.0 Hz, 1H), 4.76 (s, 2H), 4.11 (d, J = 12.0 Hz, 1H), 3.72 (d, J = 12.0 Hz, 1H). (R)-4-(Difluoromethyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 43): MS obsd. (ESI + ):340.2[(M+H) + ].

[0365] Examples 44 and 45—Compounds 44 and 45: (S)-4-hydroxy-4-isopropyl-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 44) ​​and (R)-4-hydroxy-4-isopropyl-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 45) (Stereochemistry for Compounds 44 and 45 is arbitrarily assigned) [ka] Step A: 4,4-Dimethoxy-3-(prop-1-en-2-yl)tetrahydro-2H-pyran-3-ol: [ka] To a solution of 4,4-dimethoxydihydro-2H-pyran-3(4H)-one (5.0 g, 31 mmol, 1.0 equiv., synthesized according to the procedure described in WO2013 / 152269) in anhydrous THF (100 mL) under a N2 atmosphere was added bromo(isopropenyl)magnesium (1 M, 125.0 mL, 125 mmol, 4.0 equiv.) at 0 °C. After the addition, the mixture was stirred at 0 °C for 30 min, then warmed to 25 °C and stirred for an additional 2 h under N2. The mixture was cooled to 0 °C, then quenched with saturated NH4Cl(aq) (120 mL), diluted with water (100 mL), and extracted with EtOAc (100 mL × 3). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated in vacuo. Purification by column chromatography (SiO2, 0-25% EtOAc in PE) gave 4,4-dimethoxy-3-(prop-1-en-2-yl)tetrahydro-2H-pyran-3-ol (3.5 g, 55%). 1H NMR(400MHz,CDCl3)δ ppm:5.37(m,1H),5.08-5.03(m,1H),3.76(d,J=11.4Hz,1H),3.72-3.60(m,2H),3.36(d,J=1 1.4Hz,1H),3.32(d,J=2.8Hz,6H),2.08-1.99(m,1H),1.98-1.95(m,3H),1.94-1.87(m,1H).

[0366] Step B: 3-Hydroxy-3-isopropyltetrahydro-4H-pyran-4-one: [ka] To a 1 L pressure vessel, 4,4-dimethoxy-3-(prop-1-en-2-yl)tetrahydro-2H-pyran-3-ol (17.5 g, 82.20 mmol) was added to methanol (500 mL), 4N aqueous HCl (20 mL), and Pd / C (998.34 mg, 8.22 mmol) under a nitrogen atmosphere. The vessel was sealed and shaken at room temperature under 20 atm of hydrogen pressure for 8 hours. After purging with nitrogen, the vessel was opened and the mixture was filtered through a Celite pad. The filtrate was concentrated in vacuo to give 3-hydroxy-3-isopropyltetrahydro-4H-pyran-4-one (10.9 g, 65.46 mmol, 79.63% yield, 95% purity) as a yellow liquid. MS obsd. (ESI + ):159.2[(M+H) + ].

[0367] Step C: 4-hydroxy-4-isopropyl-8-(pyridin-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] To a solution of 3-hydroxy-3-isopropyltetrahydro-4H-pyran-4-one (106 mg, 0.67 mmol, 2.0 equiv.) in toluene (10.0 mL), methyl 3-bromo-5-(pyridin-4-yl)thiophene-2-carboxylate (100 mg, 0.34 mmol, 1.0 equiv.), tris(dibenzylideneacetone)dipalladium (61 mg, 0.67 mmol, 0.20 equiv.), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (116 mg, 0.20 mmol, 0.60 equiv.), cesium carbonate (219 mg, 0.67 mmol, 2.0 equiv.), and sodium metabisulfite (13 mg, 0.070 mmol, 0.20 equiv.) were added. The mixture was stirred at 105 °C for 16 hours and then concentrated. The residue was diluted with water (20 mL) and extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (SiO2, 1-50% EtOAc in PE) to give racemic 4-hydroxy-4-isopropyl-8-(pyridin-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one. MS obsd. (ESI + ):344.4[(M+H) + ].

[0368] Step D: (S)-4-hydroxy-4-isopropyl-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 44) ​​and (R)-4-hydroxy-4-isopropyl-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 45): [ka] A solution of 4-hydroxy-4-isopropyl-8-(pyridin-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (62 mg, 0.18 mmol, 1.0 equiv.) in a mixture of NHOH (3.0 mL) and MeOH (3.0 mL) was heated in a microwave reactor at 100 °C for 3 h. The mixture was concentrated, and the residue was purified by column chromatography (SiO, 1-5% EtOAc in PE) to give racemic 4-hydroxy-4-isopropyl-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (17 mg, 28%). MS obsd. (ESI + ):343.5[(M+H) + ].

[0369] The individual enantiomers were separated via chiral SFC. (S)-4-Hydroxy-4-isopropyl-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 44): MS obsd. (ESI + ):343.2[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d6) δ ppm: 11.07 (s, 1H), 8.69 (d, J = 6.0 Hz, 2H), 8.04 (s, 1H), 7.81 (d, J = 6.0 Hz, 2H), 5.31 (s, 1H), 4.73 (s, 2H), 3.98 (d, J = 12.0 Hz, 1H), 3.55 (d, J = 11.6 Hz, 1H), 2.39-2.25 (m, 1H), 1.00 (d, J = 6.8 Hz, 3H), 0.77 (d, J = 7.2 Hz, 3H). (R)-4-Hydroxy-4-isopropyl-8-(pyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 45): MS obsd. (ESI + ):343.2[(M+H) + ].

[0370] Examples 46 and 47 - Compounds 46 and 47: (S)-4-hydroxy-4-isopropyl-8-(isothiazol-5-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 46) and (R)-4-hydroxy-4-isopropyl-8-(isothiazol-5-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 47) [ka] Racemic 4-hydroxy-4-isopropyl-8-(isothiazol-5-yl)-3,4-dihydro-1H-pyrano[4,3-b]thieno[3,2-d]pyridin-6(5H)-one was synthesized via a similar route to compounds 43 and 44). Separation via chiral SFC afforded each enantiomer. (S)-4-Hydroxy-4-isopropyl-8-(isothiazol-5-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 46): MS obsd. (ESI + ):349.0[(M+H) + ]. (R)-4-Hydroxy-4-isopropyl-8-(isothiazol-5-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 47): MS obsd. (ESI + ):349.0[(M+H) + ]; 1 H NMR(400MHz,DMSO-d6)δ ppm:11.10(s,1H),8.64(d,J=2.0Hz,1H),7.84(d,J=2.0Hz,1H),7.81(s,1H),5.31(s,1H),4.71(s,2H),3.97(d,J =11.6Hz,1H),3.54(d,J=11.6Hz,1H),2.40-2.37(m,1H),1.01-0.99(d,J=6.8Hz,3H),0.77-0.75(d,J=6.8Hz,3H).

[0371] Examples 48 and 49 - Compounds 48 and 49: (S)-8,8-difluoro-6-hydroxy-6-(methoxymethyl)-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 48) and (R)-8,8-difluoro-6-hydroxy-6-(methoxymethyl)-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 49) [ka] Step A: 5,5-Difluoro-2,2-dimethoxycyclohexan-1-one: [ka] To a solution of 5,5-difluoro-2,2-dimethoxy-cyclohexanol (29.4 g, 150 mmol, synthesized according to the procedure for Step A of Compound 12) in DCM (800.0 mL) was added Dess-Martin periodinane (95.4 g, 225 mmol) in portions over 20 minutes. The mixture was stirred for 2 hours and then concentrated. The residue was diluted with water and washed with petroleum ether:ethyl acetate (10:1, 500 mL), and the organic layer was dried over sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (SiO2, 12% EtOAc in PE) to give 5,5-difluoro-2,2-dimethoxycyclohexan-1-one (16.2 g, 55%). 1 H NMR (400MHz, CDCl3): δ ppm:3.27(s,6H),3.04(t,J=14.4Hz,2H),2.28 ppm(m,2H),2.02(m,2H).

[0372] Step B: 5,5-Difluoro-2,2-dimethoxy-1-vinylcyclohexan-1-ol: [ka] To a solution of 5,5-difluoro-2,2-dimethoxycyclohexan-1-one (1.2 g, 6.2 mmol, 1.0 equiv.) in anhydrous THF (30.0 mL) was added bromo(vinyl)magnesium (1 M, 18.5 mL, 3.0 equiv.) dropwise at 0 °C. The mixture was stirred at room temperature for 1 h, then poured into saturated NH4Cl(aq.) (10 mL) and extracted with ethyl acetate (40 mL × 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by column chromatography (SiO2, 0–10% EtOAc in PE) to give 5,5-difluoro-2,2-dimethoxy-1-vinylcyclohexan-1-ol (630 mg, 45%). 1 H NMR(400MHz,CDCl3):δ ppm:6.19(dd,J=17.2,10.8Hz,1H),5.47(dd,J=17.2,1.6Hz,1H),5.13(dd,J =10.8,1.6Hz,1H),3.24(s,3H),3.23(s,3H),2.54(s,1H),2.16-1.74(m,6H).

[0373] Step C: (((5,5-difluoro-2,2-dimethoxy-1-vinylcyclohexyl)oxy)methyl)benzene: [ka] Sodium hydride (305 mg, 7.63 mmol, 60% in mineral oil, 1.5 equiv.) was added dropwise to a solution of 5,5-difluoro-2,2-dimethoxy-1-vinylcyclohexan-1-ol (1.13 g, 5.08 mmol, 1.0 equiv.) in anhydrous DMF (30.0 mL) at 0 °C. The mixture was stirred at 0 °C for 1 h, then benzyl bromide (2.61 g, 15.2 mmol, 3.0 equiv.) was added, and the mixture was stirred at room temperature for an additional 1 h. The mixture was poured into ice water and extracted with DCM (30 mL × 2). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (SiO2, 0–10% EtOAc in PE) to give (((5,5-difluoro-2,2-dimethoxy-1-vinylcyclohexyl)oxy)methyl)benzene (1.3 g, 81%). 1 H NMR(400MHz,CDCl3):δ ppm:7.38-7.28(m,5H),6.13-6.04(m,1H),5.44(dd,J=11.2,0.8Hz,1H),5.29(d,J=18 .0Hz,1H),4.44(dd,J=34.8,11.2Hz,2H),3.42(s,3H),3.28(s,3H),2.50-1.80(m,6H).

[0374] Step D: (1-(benzyloxy)-5,5-difluoro-2,2-dimethoxycyclohexyl)methanol: [ka] Ozone was bubbled through a solution of (((5,5-difluoro-2,2-dimethoxy-1-vinylcyclohexyl)oxy)methyl)benzene (650 mg, 2.08 mmol, 1.0 equiv) in DCM (20.0 mL) at −78 °C. The mixture was stirred at −78 °C for 30 min and then warmed to room temperature. MeOH (5.0 mL) and NaBH (236 mg, 6.24 mmol, 3.0 equiv) were added, and the mixture was stirred for 5 min. The mixture was poured into water and extracted with ethyl acetate (70 mL × 2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (15–30% EtOAc in SiO2PE) to give (1-(benzyloxy)-5,5-difluoro-2,2-dimethoxycyclohexyl)methanol (1.1 g, 83%). 1 H NMR(400MHz,CDCl3):δ ppm:7.36-7.19(m,5H),4.53(d,J=1.6Hz,2H),3.92(d,J=12.2Hz,1H),3.6 9-3.57(m,1H),3.38(s,3H),3.25(s,3H),2.35(m,2H),2.08-1.85(m,4H).

[0375] Step E: (((5,5-difluoro-2,2-dimethoxy-1-(methoxymethyl)cyclohexyl)oxy)methyl)benzene: [ka] Sodium hydride (265 mg, 6.64 mmol, 60% in mineral oil, 1.4 equiv.) was added dropwise to a solution of (1-(benzyloxy)-5,5-difluoro-2,2-dimethoxycyclohexyl)methanol (1.5 g, 4.74 mmol, 1.0 equiv.) in anhydrous DMF (30.0 mL) at 0 °C. The mixture was stirred at room temperature for 1 h, and then iodomethane (6.73 g, 47.4 mmol, 10 equiv.) was added. The mixture was stirred for 2 h, then poured into ice water and extracted with DCM (60 mL × 2). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (SiO2, 0–10% EtOAc in PE) to give (((5,5-difluoro-2,2-dimethoxy-1-(methoxymethyl)cyclohexyl)oxy)methyl)benzene (1.2 g, 76%). 1 H NMR(400MHz,CDCl3):δ ppm:7.38-7.17(m,5H),4.72(s,2H),3.74(s,2H),3.39(s,3H),3.34(s,3H),3.30(s,3H),2.64-2.25(m,3H),2.22-1.75(m,3H).

[0376] Step F: 2-(benzyloxy)-4,4-difluoro-2-(methoxymethyl)cyclohexan-1-one: [ka] A mixture of (((5,5-difluoro-2,2-dimethoxy-1-(methoxymethyl)cyclohexyl)oxy)methyl)benzene (1.33 g, 4.03 mmol, 1.0 equiv), I2 (102 mg, 402 μmol, 0.1 equiv), and acetone (30.0 mL) was stirred at room temperature for 2 h. The mixture was poured into water, quenched with saturated Na2SO3 (aq), and extracted with DCM (60 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (SiO2, 0–5% EtOAc in PE) to give 2-(benzyloxy)-4,4-difluoro-2-(methoxymethyl)cyclohexan-1-one (1.05 g, 91%). 1 H NMR(400MHz,CDCl3):δ ppm:7.42-7.26(m,5H),4.60(dd,J=11.4Hz,1H),4.40(dd,J=11.4Hz,1H),3.68(m,2H),3.37(s,3H),2.92-2.62(m,2H),2.52-2.23(m,4H).

[0377] Step G: 6-(benzyloxy)-8,8-difluoro-6-(methoxymethyl)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one: [ka] A mixture of methyl 3-bromo-5-[1-(2-trimethylsilylethoxymethyl)pyrazol-4-yl]thiophene-2-carboxylate (660 mg, 1.6 mmol, 1.8 equiv.), 2-(benzyloxy)-4,4-difluoro-2-(methoxymethyl)cyclohexan-1-one (250 mg, 880 μmol, 1.0 equiv.), sodium metabisulfite (50 mg, 263 μmol, 0.3 equiv.), cesium carbonate (859 mg, 2.64 mmol, 3.0 equiv.), Sphos-Pd-G3 (137 mg, 175 μmol, 0.20 equiv.), and toluene (50.0 mL) was stirred at 105° C. under nitrogen for 16 hours. The mixture was poured into water and extracted with EtOAc (60 mL × 2). The combined organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (SiO, 0-35% EtOAc in PE) to give 6-(benzyloxy)-8,8-difluoro-6-(methoxymethyl)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (360 mg, 69%). MS obsd. (ESI + ):589.5[(M+H) + ].

[0378] Step H: 6-(benzyloxy)-8,8-difluoro-6-(methoxymethyl)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one: [ka] A mixture of 6-(benzyloxy)-8,8-difluoro-6-(methoxymethyl)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (180 mg, 305 μmol, 1.0 equiv.), ammonium hydroxide (8.0 mL), and 2-propyl alcohol (8.0 mL) was stirred at 100° C. in a microwave reactor for 4 hours. The mixture was concentrated in vacuo and the residue was purified by column chromatography (SiO, 0-1% MeOH in DCM) to give 6-(benzyloxy)-8,8-difluoro-6-(methoxymethyl)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (80 mg, 44%). MS obsd. (ESI + ):588.5[(M+H) + ].

[0379] Step I: (S)-8,8-difluoro-6-hydroxy-6-(methoxymethyl)-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 48) and (R)-8,8-difluoro-6-hydroxy-6-(methoxymethyl)-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 49): [ka] Boron trichloride (1 M, 4.7 mL, 15 equiv) was added to a solution of 6-(benzyloxy)-8,8-difluoro-6-(methoxymethyl)-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (190 mg, 320 μmol, 1.0 equiv) in DCM (25.0 mL) at 0° C. The mixture was stirred at room temperature for 2 h, then quenched with MeOH, concentrated, and neutralized with NH / MeOH (7 M). The residue was purified by column chromatography (SiO, 0-4% MeOH in DCM) to give racemic 8,8-difluoro-6-hydroxy-6-(methoxymethyl)-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (64 mg, 53%). MS obsd. (ESI + ):368.0[(M+H) + ].

[0380] Separation via SFC afforded each enantiomer. (S)-8,8-Difluoro-6-hydroxy-6-(methoxymethyl)-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 48): MS obsd. (ESI + ):368.0[(M+H) + ]. (R)-8,8-Difluoro-6-hydroxy-6-(methoxymethyl)-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 49): MS obsd. (ESI + ):368.0[(M+H) + ]; 1 H NMR(400MHz,DMSO-d6)δ ppm:13.23(s,1H),10.70(s,1H),8.30(s,1H),7.97(s,1H),7.58(s,1H),5.69(s,1H),3.60(d d,J=45.2,10.0Hz,2H),3.29(s,3H),3.21-3.10(m,2H),2.77-2.61(m,1H),2.38-2.19(m,1H).

[0381] Examples 50 and 51 - Compounds 50 and 51: (S)-6-ethyl-8,8-difluoro-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 50) and (R)-6-ethyl-8,8-difluoro-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 51) [ka] Step A: 1-Ethyl-5,5-difluoro-2,2-dimethoxycyclohexan-1-ol: [ka] To a solution of 5,5-difluoro-2,2-dimethoxy-1-vinylcyclohexan-1-ol (1.8 g, 8.1 mmol, 1.0 equiv) in MeOH (90 mL) and HO (2.0 mL) was added wet Pd / C (10%, 540 mg). The mixture was purged with nitrogen and then stirred under a hydrogen atmosphere at 50 °C for 16 h. The reaction mixture was cooled to room temperature, filtered through Celite, and the filter cake was washed with MeOH (100 mL). The filtrate was concentrated in vacuo and purified by flash column chromatography (SiO, 10–50% EtOAc in PE) to give 1-ethyl-5,5-difluoro-2,2-dimethoxycyclohexan-1-ol (1.5 g, 82%). 1 H NMR(400MHz,CDCl3)δ ppm:3.39(s,3H),3.33(s,3H),1.99-1.62(m,8H),0.99(t,J=7.6Hz,3H).

[0382] Step B: 2-Ethyl-4,4-difluoro-2-hydroxycyclohexan-1-one: [ka] To a solution of 1-ethyl-5,5-difluoro-2,2-dimethoxycyclohexan-1-ol (1.8 g, 8.03 mmol, 1.0 equiv) in DCM (28.0 mL) was added water (7.0 mL) and trifluoroacetic acid (7.0 mL). The mixture was stirred at room temperature for 1 h and then concentrated. Purification by column chromatography (SiO2, 10–50% EtOAc in PE) gave 2-ethyl-4,4-difluoro-2-hydroxycyclohexan-1-one (1.2 g, 85%). 1 H NMR(400MHz,CDCl3)δ ppm:2.84-2.70(m,2H),2.59-2.46(m,2H),2.29-2.11(m,2H),1.97-1.87(m,1H),1.84-1.74(m,1H),0.84(t,J=7.2Hz,3H).

[0383] Step C: 6-Ethyl-8,8-difluoro-6-hydroxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one: [ka] To a solution of 2-ethyl-4,4-difluoro-2-hydroxycyclohexan-1-one (205 mg, 1.15 mmol, 2.0 equiv.) and methyl 3-bromo-5-[1-(2-trimethylsilylethoxymethyl)pyrazol-4-yl]thiophene-2-carboxylate (240 mg, 0.58 mmol, 1.0 equiv.) in toluene (15.0 mL), sodium metabisulfite (22 mg, 0.12 mmol, 0.2 equiv.), CsCO (374 mg, 1.15 mmol, 2.0 equiv.), and Pd(dba) (105 mg, 0.12 mmol, 0.2 equiv.), Xantphos (100 mg, 0.17 mmol, 0.3 equiv.) were added under a N atmosphere. The mixture was stirred at 105 °C for 16 h, then filtered and concentrated in vacuo. The residue was purified by column chromatography (SiO, 0-100% EtOAc in PE) to give 6-ethyl-8,8-difluoro-6-hydroxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (224 mg, 80%). MS obsd. (ESI + ):483.7[(M+H) + ].

[0384] Step D: 6-Ethyl-8,8-difluoro-6-hydroxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one: [ka] A solution of 6-ethyl-8,8-difluoro-6-hydroxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (595 mg, 1.23 mmol, 1.0 equiv) in a mixture of MeOH (9 mL) and NH4OH (9.0 mL) was heated in a microwave reactor at 100 °C for 3 h. The mixture was concentrated, and the residue was purified by column chromatography (SiO, 0-10% MeOH in DCM) to give 6-ethyl-8,8-difluoro-6-hydroxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (360 mg, 60%). MS obsd. (ESI + ):482.7[(M+H) + ].

[0385] Step E: (S)-6-ethyl-8,8-difluoro-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 50) and (R)-6-ethyl-8,8-difluoro-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 51): [ka] To a solution of 6-ethyl-8,8-difluoro-6-hydroxy-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (357 mg, 0.74 mmol, 1.0 equiv.) in DCM (5.0 mL) was added trifluoroacetic acid (5.0 mL). The mixture was stirred at room temperature for 1 h, then concentrated in vacuo, and the residue was purified by column chromatography (SiO, 0-10% MeOH in DCM) to give racemic 6-ethyl-8,8-difluoro-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (140 mg, 54%). MS obsd. (ESI + ):352.4[(M+H) + ].

[0386] The individual isomers were separated by chiral SFC. (S)-6-Ethyl-8,8-difluoro-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 50): MS obsd. (ESI + ):352.2[(M+H) + ]. (R)-6-Ethyl-8,8-difluoro-6-hydroxy-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 51): MS obsd. (ESI + ):352.0[(M+H) + ]; 1 H NMR(400MHz,DMSO-d6)δ ppm:13.23(s,1H),10.74(s,1H),8.30(s,1H),8.00(s,1H),7.57(s,1H),5.33(s,1H),3.30-3.27(m, 2H),2.68-2.54(m,1H),2.33-2.20(m,1H),1.96-1.87(m,1H),1.83-1.74(m,1H),0.89-0.84(m,3H).

[0387] Examples 52 and 53—Compounds 52 and 53: (S)-8,8-difluoro-6-methoxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 52) and (R)-8,8-difluoro-6-methoxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 53) (stereoisomers arbitrarily assigned) [ka] Step A: 5,5-Difluoro-2,2-dimethoxy-1-methylcyclohexan-1-ol: [ka] To a dry three-neck flask containing 5,5-difluoro-2,2-dimethoxycyclohexan-1-one (5.65 g, 29.1 mmol, 1.0 equiv.) in anhydrous THF (270.0 mL) was added methylmagnesium bromide (3 M in diethyl ether, 29.1 mL, 87.4 mmol, 3.0 equiv.) at 0 °C. The mixture was stirred at room temperature under a N atmosphere for 2 h and monitored by TLC until completion. The mixture was quenched with saturated NH4Cl (aq.) at 0 °C, diluted with water (125 mL), and extracted with EtOAc (90 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and the residue was purified by column chromatography (SiO2, 0–20% EtOAc in PE) to give 5,5-difluoro-2,2-dimethoxy-1-methylcyclohexan-1-ol (5.98 g, 97%). 1 H NMR(400MHz,CDCl3)δ ppm:3.41(s,3H),3.32(s,3H),2.69(s,1H),2.20-2.06(m,2H),2.03-1.90(m,2H),1.85-1.68(m,2H),1.32(s,3H).

[0388] Step B: 4,4-Difluoro-1,1,2-trimethoxy-2-methylcyclohexane: [ka] To a dry three-neck round-bottom flask containing sodium hydride (2.28 g, 56.7 mmol, 60% in mineral oil, 2.0 equiv.) was added anhydrous THF (170.0 mL) at 0° C., followed by a mixture of 5,5-difluoro-2,2-dimethoxy-1-methylcyclohexan-1-ol (5.98 g, 28.5 mmol, 1.0 equiv.) in THF (3.0 mL). The mixture was stirred at 0° C. for 30 minutes, then iodomethane (40.4 g, 285 mmol, 10.0 equiv.) was added, and the mixture was stirred for an additional 1.5 hours. Additional iodomethane (40.4 g, 285 mmol, 10.0 equiv.) was added to the reaction mixture at 0° C. The mixture was stirred for an additional 1.5 hours, then water was added at 0° C. The mixture was extracted with EtOAc (160 mL × 2), and the combined organic layers were washed with brine (150 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. Purification by column chromatography (SiO2, 0–10% EtOAc in PE) gave 4,4-difluoro-1,1,2-trimethoxy-2-methylcyclohexane (5.1 g, 80%). 1 H NMR(400MHz,CDCl3)δ ppm:3.37(s,3H),3.33(s,3H),3.27(s,3H),2.27-2.20(m,1H),2.18-2.05(m,2H),2.00-1.86(m,2H),1.80-1.69(m,1H),1.29(s,3H).

[0389] Step C: 4,4-Difluoro-2-methoxy-2-methylcyclohexan-1-one: [ka] To a flask containing 4,4-difluoro-1,1,2-trimethoxy-2-methylcyclohexane (2.58 g, 11.51 mmol) in DCM (28 mL) was added water (7 mL) and trifluoroacetic acid (7 mL) at 0 °C. The mixture was stirred at room temperature for 5 hours and then extracted with DCM (30 mL × 2). The combined organic layer was concentrated in vacuo. The residue was diluted with DCM (20 mL) and washed with aqueous NaHCO3 (10 mL × 2), and the organic layer was dried over anhydrous sodium sulfate. The mixture was filtered and concentrated in vacuo to give 4,4-difluoro-2-methoxy-2-methylcyclohexan-1-one (1.9 g, 10.1 mmol, 88%). 1 H NMR(400MHz,CDCl3)δ ppm:3.21(s,3H),2.91-2.85(m,1H),2.65-2.57(m,1H),2.48-2.41(m,2H),2.23-2.09(m,2H),1.30(s,3H).

[0390] Step D: 8,8-difluoro-6-methoxy-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one: [ka] To a solution of methyl 3-bromo-5-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)thiophene-2-carboxylate (1.0 g, 2.4 mmol) and 4,4-difluoro-2-methoxy-2-methylcyclohexan-1-one (598 mg, 3.35 mmol) in toluene (30.0 mL) was added CsCO (2.34 g, 7.19 mmol), Pd(dba) (439 mg, 479 μmol), and NaSO (91.1 mg, 479 μmol). The mixture was stirred at 105 °C under nitrogen for 12 hours. The mixture was cooled to 0 °C, HO (40 mL) was added, and the resulting mixture was extracted with EtOAc (30 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by column chromatography (SiO, 0-10% EtOAc in PE) gave 8,8-difluoro-6-methoxy-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (385 mg, 33%). MS obsd. (ESI + ):483.4[(M+H) + ].

[0391] Step E: 8,8-difluoro-6-methoxy-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one: [ka] To a solution of 8,8-difluoro-6-methoxy-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (0.70 g, 1.45 mmol) in MeOH (6.0 mL) was added ammonium hydroxide (6.0 mL). The mixture was stirred at 95 °C in a microwave reactor for 2 hours. The mixture was cooled to 0 °C, and HO (20 mL) was added. The resulting mixture was extracted with EtOAc (50 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash column chromatography (SiO, 0-30% EtOAc in PE) to give 8,8-difluoro-6-methoxy-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (170 mg, 24%). MS obsd. (ESI + ):482.5[(M+H) + ].

[0392] Step F: (S)-8,8-difluoro-6-methoxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 52) and (R)-8,8-difluoro-6-methoxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 53): [ka] To a solution of 8,8-difluoro-6-methoxy-6-methyl-2-[1-(2-trimethylsilylethoxymethyl)pyrazol-4-yl]-7,9-dihydro-5H-thieno[2,3-c]quinolin-4-one (170 mg, 353 μmol, 1.0 equiv.) in DCM (5.0 mL) was added trifluoroacetic acid (402 mg, 3.53 mmol, 272 μL, 10.0 equiv.) at 0° C. The mixture was stirred at 25° C. for 15 hours. The mixture was then cooled to 0° C., and saturated aqueous NaHCO (20 mL) was added dropwise at 0° C. The resulting mixture was extracted with EtOAc (20 mL × 3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash column chromatography (SiO, 0–10% MeOH in DCM) to give racemic 8,8-difluoro-6-methoxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (100 mg, 80%).

[0393] The individual enantiomers were isolated via chiral SFC. (S)-8,8-Difluoro-6-methoxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 52): MS obsd. (ESI + ):352.1[M+H] + (R)-8,8-Difluoro-6-methoxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 53): MS obsd. (ESI + ):352.1[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ ppm:13.23(s,1H),11.02(s,1H),8.28(s,1H),8.01(s,1H),7.59(s,1H),3.44 -3.37(m,2H),3.07(s,3H)2.82-2.61(m,1H),2.40-2.26(m,1H),1.56(s,3H).

[0394] Examples 54 and 55 - Compounds 54 and 55: (R)-8,8-difluoro-6-hydroxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 54) and (S)-8,8-difluoro-6-hydroxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (Compound 55) (stereoisomers arbitrarily assigned) [ka] Step A: (((5,5-difluoro-2,2-dimethoxy-1-methylcyclohexyl)oxy)methyl)benzene: [ka] To a solution of 5,5-difluoro-2,2-dimethoxy-1-methylcyclohexan-1-ol (300 mg, 1.43 mmol, 1.0 equiv) in anhydrous DMF (3.0 mL) was added NaH (109 mg, 2.85 mmol, 60% in mineral oil, 2.0 equiv) at 0 °C. The mixture was stirred at 0 °C for 10 minutes, and then benzyl bromide (488.16 mg, 2.85 mmol, 339 μL, 2.0 equiv) was added dropwise. The mixture was stirred at room temperature for 1 hour, then quenched with saturated aqueous NH4Cl and diluted with water (60 mL). The mixture was extracted with EtOAc (40 mL × 3), and the combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (SiO2, 0-4% EtOAc in PE) to give (((5,5-difluoro-2,2-dimethoxy-1-methylcyclohexyl)oxy)methyl)benzene (370 mg, 86%). 1H NMR(400MHz,CDCl3)δ ppm:7.36-7.24(m,5H),4.60-4.52(m,2H),3.39(s,3H),3.33(s,3H),2.50-2.35(m,1H), 2.26-2.22(m,1H),2.15-2.07(m,1H),2.05-1.89(m,2H),1.83-1.77(m,1H),1.38(s,3H).

[0395] Step B: 2-(benzyloxy)-4,4-difluoro-2-methylcyclohexan-1-one: [ka] To a solution of (((5,5-difluoro-2,2-dimethoxy-1-methylcyclohexyl)oxy)methyl)benzene (370 mg, 1.23 mmol, 1.0 equiv) in acetone (15.0 mL) was added iodine (31.7 mg, 125 μmol, 0.10 equiv). The mixture was stirred at room temperature for 10 min, then quenched with saturated NaSO (aq) (20 mL) and extracted with DCM (30 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (SiO, 0–10% EtOAc in PE) to give 2-(benzyloxy)-4,4-difluoro-2-methylcyclohexan-1-one (280 mg, 88%). 1 H NMR(400MHz,CDCl3)δ ppm:7.35-7.26(m,5H),4.56(d,J=11.6Hz,1H),4.26(d,J=11.6Hz,1H),2.94-2. 91(m,1H),2.79-2.74(m,1H),2.48-2.42(m,2H),2.25-2.18(m,2H),1.41(s,3H).

[0396] Step C: 6-(benzyloxy)-8,8-difluoro-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one: [ka] To a solution of methyl 3-bromo-5-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)thiophene-2-carboxylate (240 mg, 575 μmol, 1.0 equiv.) and 2-(benzyloxy)-4,4-difluoro-2-methylcyclohexan-1-one (292 mg, 1.15 mmol, 2.0 equiv.) in toluene (18.0 mL) was added Pd(dba) (105 mg, 115 μmol, 0.20 equiv.), Xantphos (133 mg, 230 μmol, 0.40 equiv.), and cesium carbonate (562 mg, 1.72 mmol, 3.0 equiv.). The mixture was stirred under nitrogen at 105° C. for 16 h, then cooled and filtered. The filtrate was concentrated in vacuo, and the residue was purified by column chromatography (SiO, 0-16% EtOAc in PE) to give 6-(benzyloxy)-8,8-difluoro-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (220 mg, 68%). MS obsd. (ESI + ):559.5[(M+H) + ].

[0397] Step D: 6-(benzyloxy)-8,8-difluoro-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one: [ka] To a solution of 6-(benzyloxy)-8,8-difluoro-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydro-4H-thieno[2,3-c]chromen-4-one (220 mg, 394 μmol) in MeOH (7 mL) was added ammonia (7 M in MeOH, 7.0 mL). The mixture was stirred at 95° C. for 16 hours, then cooled and concentrated in vacuo. The residue was purified by column chromatography (SiO, 0-5% MeOH in DCM) to give 6-(benzyloxy)-8,8-difluoro-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (180 mg, 77%). MS obsd. (ESI + ):558.6[(M+H) + ].

[0398] Step E: (R)-8,8-difluoro-6-hydroxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 54) and (S)-8,8-difluoro-6-hydroxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 55) (stereoisomers arbitrarily assigned): [ka] To a solution of 6-(benzyloxy)-8,8-difluoro-6-methyl-2-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (90 mg, 161 μmol, 1.0 equiv.) in DCM (10.0 mL) was added boron trichloride (1 M, 2.0 mL, 12 equiv.) at −5° C. The mixture was stirred at −5° C. for 1.5 h, quenched with MeOH, and then concentrated under a stream of nitrogen. The pH was adjusted to approximately 9 by slow addition of NH / MeOH (7 M). The mixture was then concentrated in vacuo, and the residue was purified by flash column chromatography (SiO, 0–10% MeOH in DCM) to give racemic 8,8-difluoro-6-hydroxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (23 mg, 42%). MS obsd. (ESI + ):338.3[(M+H) + ].

[0399] The individual enantiomers were separated via chiral SFC. (R)-8,8-Difluoro-6-hydroxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 54): MS obsd. (ESI + ):338.0[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d6) δ ppm: 13.24 (s, 1H), 10.88 (s, 1H), 8.13 (s, 2H), 7.56 (s, 1H), 5.57 (s, 1H), 3.33-3.24 (m, 2H), 2.50-2.42 (m, 2H), 1.54 (s, 3H). (S)-8,8-Difluoro-6-hydroxy-6-methyl-2-(1H-pyrazol-4-yl)-6,7,8,9-tetrahydrothieno[2,3-c]quinolin-4(5H)-one (compound 55): MS obsd. (ESI + ):338.0[(M+H) + ].

[0400] Examples 56 and 57 - Compounds 56 and 57: (S)-4-(2-hydroxypropan-2-yl)-4-methyl-8-(1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (Compound 56) and (R)-4-(2-hydroxypropan-2-yl)-4-methyl-8-(1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (Compound 57) [ka] Step A: 4-methyl-6-oxo-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,4,5,6-tetrahydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridine-4-carbaldehyde: [ka] A mixture of 4-(hydroxymethyl)-4-methyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (100 mg, 223 μmol, 1.0 equiv., synthesized according to the procedure for compounds 34 and 35), IBX (626 mg, 2.23 mmol, 10.0 equiv.), and DCM (12 mL) was stirred at 45° C. for 24 hours. The mixture was quenched with saturated aqueous NaSO and extracted with DCM (30 mL × 3). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (SiO, 0-4% MeOH in DCM) to give 4-methyl-6-oxo-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,4,5,6-tetrahydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridine-4-carbaldehyde (80 mg, 80%). MS obsd. (ESI +):464.3[(M+H2O+H) + ].

[0401] Step B: 4-(1-hydroxyethyl)-4-methyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one: [ka] To a solution of 4-methyl-6-oxo-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,4,5,6-tetrahydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridine-4-carbaldehyde (160 mg, 360 μmol, 1.0 equiv.) in THF (7.5 mL) was added methylmagnesium bromide (3 M in diethyl ether, 23.3 mL, 195 equiv.). The mixture was stirred at room temperature for 1.5 h, quenched with saturated NH4Cl (aq.), then poured into water and extracted with DCM (50 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (SiO, 0-4% MeOH in DCM) to give 4-(1-hydroxyethyl)-4-methyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (152 mg, 82%). MS obsd. (ESI + ):462.3[(M+H) + ].

[0402] Step C: 4-acetyl-4-methyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one: [ka] To a solution of 4-(1-hydroxyethyl)-4-methyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (126 mg, 273 μmol, 1.0 equiv) in DCM (40.0 mL) was added Dess-Martin periodinane (694 mg, 1.64 mmol, 6.0 equiv) in portions at 0° C. The mixture was stirred at room temperature for 1.5 h, then quenched with saturated NaSO (aq) and extracted with DCM (30 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (SiO, 0-3% MeOH in DCM) to give 4-acetyl-4-methyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (85 mg, 67%). MS obsd. (ESI + ):460.3[(M+H) + ].

[0403] Step D: 4-(2-hydroxypropan-2-yl)-4-methyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one: [ka] To a solution of 4-acetyl-4-methyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (85 mg, 185 μmol, 1.0 equiv.) in THF (2.0 mL) was added methylmagnesium bromide (3 M in diethyl ether, 2.0 mL, 32.4 equiv.). The mixture was stirred at room temperature for 2 hours, quenched with saturated NH4Cl (aq.), then poured into water and extracted with DCM (30 mL × 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash column chromatography (SiO, 0-4% MeOH in DCM) to give 4-(2-hydroxypropan-2-yl)-4-methyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (50 mg, 56%). MS obsd. (ESI + ):476.3[(M+H) + ].

[0404] Step E: (S)-4-(2-hydroxypropan-2-yl)-4-methyl-8-(1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (compound 56) and (R)-4-(2-hydroxypropan-2-yl)-4-methyl-8-(1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (compound 57): [ka] To a solution of 4-(2-hydroxypropan-2-yl)-4-methyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (75 mg, 160 μmol, 1.0 equiv) in DCM (6.0 mL) was added trifluoroacetic acid (2.0 mL) at 0° C. The mixture was warmed to room temperature and stirred for 4 h. The mixture was concentrated under a stream of nitrogen and the pH was adjusted to approximately 9 by slow addition of NH / MeOH (7 M) at 0° C. The mixture was concentrated to give a residue that was purified by column chromatography (SiO, 0-7% MeOH in DCM) to give racemic 4-(2-hydroxypropan-2-yl)-4-methyl-8-(1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (48 mg, 85%). MS obsd. (ESI + ):346.0[(M+H) + ].

[0405] The individual enantiomers were separated via chiral SFC. (S)-4-(2-hydroxypropan-2-yl)-4-methyl-8-(1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (compound 56): MS obsd. (ESI + ):346.2[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d6) δ ppm: 13.22 (s, 1H), 10.21 (s, 1H), 8.31 (s, 1H), 7.97 (s, 1H), 7.51 (s, 1H), 5.98 (s, 1H), 4.02-3.89 (m, 2H), 2.76-2.60 (m, 2H), 1.53 (s, 3H), 1.27 (s, 3H), 1.00 (s, 3H). (R)-4-(2-hydroxypropan-2-yl)-4-methyl-8-(1H-pyrazol-4-yl)-1,5-dihydro-2H-pyrano[3,4-b]thieno[3,2-d]pyridin-6(4H)-one (compound 57). MS obsd. (ESI +):346.2[(M+H) + ].

[0406] Example 58 - Compound 58: 4,4-dimethyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one [ka] Step A: 4,4-dimethyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] Methyl 3-bromo-5-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)thiophene-2-carboxylate (100 mg, 240 μmol, 1.0 equiv.) was dissolved in DMF (10.0 mL), followed by the addition of 3,3-dimethyltetrahydropyran-4-one (61 mg, 480 μmol, 2.0 equiv.), NaSO (9.0 mg, 48 μmol, 0.20 equiv.), CsCO (230 mg, 720 μmol, 3.0 equiv.), and BINAP-Pd-G (24 mg, 24 μmol, 0.10 equiv.). The mixture was stirred at 105 °C for 16 h, cooled, filtered, and concentrated. Purification by column chromatography (0-4% MeOH in SiO2 DCM) gave 4,4-dimethyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (20 mg, 19%). MS obsd. (ESI + ):433.4[(M+H) + ].

[0407] Step B: 4,4-dimethyl-8-(1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] A mixture of 4,4-dimethyl-8-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (45 mg, 100 μmol), DCM (5.0 mL), and trifluoroacetic acid (1.5 mL) was stirred at room temperature for 2 hours. The mixture was concentrated in vacuo to give 4,4-dimethyl-8-(1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (40 mg), which was used without further purification. MS obsd. (ESI + ):303.2[(M+H) + ].

[0408] Step C: 4,4-dimethyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one: [ka] A mixture of 4,4-dimethyl-8-(1H-pyrazol-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (40 mg, crude), NHOH (5.0 mL), and MeOH (5.0 mL) was heated to 100 °C in a microwave reactor for 4 h. The mixture was concentrated in vacuo, and the residue was purified by column chromatography (SiO, 0-5% MeOH in DCM) to give 4,4-dimethyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (9.0 mg, 36%). MS obsd. (ESI + ):302.2[(M+H)+ ]; 1 H NMR(400MHz,DMSO-d6)δ ppm:13.21(brs,1H),11.20(s,1H),8.29(s,1H),7.94(s,1H),7.37(s,1H),4.69(s,2H),3.57(s,2H),1.26(s,6H).

[0409] Examples 59 and 60—Compounds 59 and 60: (S)-4-(tert-butyl)-8-(3-fluoropyridin-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 59) and (R)-4-(tert-butyl)-8-(3-fluoropyridin-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 60) (Stereochemistry for Compounds 59 and 60 is arbitrarily assigned) [ka] Step A: 4,4-Dimethoxytetrahydro-2H-pyran-3-ol: [ka] A solution of tetrahydropyran-4-one (40.0 g, 400 mmol, 1.0 equiv) and potassium hydroxide (67.3 g, 1.20 mol, 3.0 equiv) in MeOH (1.7 L) was cooled to 0 °C, and I2 (117 g, 459 mmol, 1.15 equiv) was slowly added. The reaction mixture was allowed to warm to room temperature, and after 4 h, the mixture was concentrated. The crude product was triturated with EtOAc (300 mL) to give 4,4-dimethoxytetrahydro-2H-pyran-3-ol (50 g, used without further purification). 1H NMR(400MHz,DMSO-d6)δ 4.75(s,1H),3.68-3.54(m,2H),3.52-3.41(m,2H),3.32-3.25(m,1H),3.12(s,3H),3.11(s,3H),1.84-1.76(m,1H),1.62-1.57(m,1H).

[0410] Step B: 4,4-Dimethoxydihydro-2H-pyran-3(4H)-one: [ka] A solution of 4,4-dimethoxytetrahydropyran-3-ol (40.0 g, 247 mmol, 1.0 equiv) in DCM (900 mL) was cooled to 0 °C, and DMP (157 g, 370 mmol, 1.5 equiv) was added. The mixture was heated to 30 °C for 16 h, after which a mixture of petroleum ether and EtOAc (8:1, 400 mL) was added. The filtrate was concentrated and purified (SiO, 5% petroleum ether in EtOAc) to give 4,4-dimethoxydihydro-2H-pyran-3(4H)-one (23 g, 45%, 2 steps) as a pale yellow solid.

[0411] 1 H NMR(400MHz,DMSO-d6)δ 3.99(s,2H),3.87-3.80(m,2H),3.14(s,6H),2.16-2.10(m,2H).

[0412] Step C: 3-(tert-butyl)-3-hydroxytetrahydro-4H-pyran-4-one: [ka] A solution of 4,4-dimethoxytetrahydropyran-3-one (10.0 g, 62.4 mmol, 1.0 equiv) in anhydrous THF (100 mL) was cooled to −78 °C, and t-BuLi (1.3 M in THF, 62 mL, 1.3 equiv) was added dropwise over 30 min. The mixture was stirred at −78 °C for 2 h, after which HCl (2 M aqueous solution, 94 mL) was added dropwise over 15 min. After addition, the mixture was stirred at 0 °C for 2 h. The mixture was extracted with EtOAc (3 × 100 mL), washed with brine (2 × 100 mL), and the combined organic layers were concentrated and purified (SiO, 0–10% EtOAc in PE) to give 3-(tert-butyl)-3-hydroxytetrahydro-4H-pyran-4-one (3.6 g, 33%). 1 H NMR(400MHz,DMSO-d6)δ 5.05(s,1H),4.06(dd,J=12.0,1.2Hz,1H),4.03-3.95(m,1H),3.75-3.64(m,1H),3.35(d,J=12.0Hz,1H),2.60-2.52(m,2H),0.95(s,9H).

[0413] Step D: 4-(tert-butyl)-4-hydroxy-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] To a dry three-neck flask containing methyl 3-bromothiophene-2-carboxylate (2 g, 9.05 mmol, 1.0 equiv.) and 3-tert-butyl-3-hydroxy-tetrahydropyran-4-one (1.56 g, 9.05 mmol, 1.0 equiv.), anhydrous toluene (50 mL) was added. The mixture was degassed and purged with N (3x), followed by the addition of Pd(dba) (414 mg, 452 μmol, 0.050 equiv.), Xantphos (524 mg, 905 μmol, 0.10 equiv.), and KPO (3.84 g, 18.1 mmol, 2.0 equiv.). It was degassed and purged again with N (3x), and stirred at 105 °C for 16 h. The mixture was cooled to room temperature, filtered, and the filtrate was concentrated. Purification by column chromatography (SiO, 0-30% EtOAc in PE) and further purification by reverse-phase column chromatography (C18 SiO, 0-30% MeCN in water (0.1% NH4OH) gave 4-(tert-butyl)-4-hydroxy-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (600 mg, 2.14 mmol, 24%) as a white solid. MS obsd. (ESI + ):281.2[(M+H) + ].

[0414] Step E: (4-(tert-butyl)-4-hydroxy-6-oxo-4,6-dihydro-1H,3H-pyrano[4,3-b]thieno[3,2-d]pyran-8-yl)boronic acid: [ka] A mixture of 4-(tert-butyl)-4-hydroxy-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (500 mg, 1.78 mmol, 1.0 equiv), 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1.37 g, 10.7 mmol, 1.55 mL, 6.0 equiv), (1,5-cyclooctadiene)(methoxy)iridium dimer (59.1 mg, 89.2 μmol, 0.050 equiv), 2,6-diisopropyl-N-(pyridin-2-ylmethylene)aniline (47.5 mg, 178 μmol, 0.10 equiv) and anhydrous DME (10 mL) was stirred at 85° C. for 24 hours. The mixture was cooled and concentrated in vacuo to give (4-(tert-butyl)-4-hydroxy-6-oxo-4,6-dihydro-1H,3H-pyrano[4,3-b]thieno[3,2-d]pyran-8-yl)boronic acid (2 g, crude), which was used without further purification. MS obsd. (ESI + ):325.5[(M+H) + ].

[0415] Step F: 10-tert-butyl-4-(3-fluoro-4-pyridyl)-10-hydroxy-8,12-dioxa-5-thiatricyclo[7.4.0.0 2,6 ]Trideca-1(9),2(6),3-trien-7-one: [ka] (10-tert-butyl-10-hydroxy-7-oxo-8,12-dioxa-5-thiatricyclo[7.4.0.0]) in THF (12 mL) and HO (1.2 mL) 2,6To a mixture of ]trideca-1(9),2(6),3-trien-4-yl)boronic acid (130 mg, 401 μmol, 1.0 equiv.), NaCO (128 mg, 1.20 mmol, 3.0 equiv.), 3-fluoro-4-iodo-pyridine (99 mg, 441 μmol, 1.1 equiv.), and Sphos-Pd-G (32 mg, 40.1 μmol, 0.10 equiv.) were added. The mixture was degassed and purged with N (2×) and then stirred at 50 °C for 6 h. The reaction mixture was concentrated, and the residue was diluted with DCM / MeOH = 10 / 1 (300 mL). The mixture was filtered, and the filtrate was concentrated in vacuo. The crude residue was purified by flash column chromatography (0-5% MeOH in SiO2 DCM) to give 10-tert-butyl-4-(3-fluoro-4-pyridyl)-10-hydroxy-8,12-dioxa-5-thiatricyclo[7.4.0.0 2,6 ] to give trideca-1(9),2(6),3-trien-7-one (280 mg, 23%) as a yellow solid. MS obsd. (ESI + ):376.4[(M+H) + ].

[0416] Step G: (S)-4-(tert-butyl)-8-(3-fluoropyridin-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 59) and (R)-4-(tert-butyl)-8-(3-fluoropyridin-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 60): [ka] 10-tert-Butyl-4-(3-fluoro-4-pyridyl)-10-hydroxy-8,12-dioxa-5-thiatricyclo[7.4.0.0] in NH3 / MeOH (7.0 M, 10 mL) 2,6A solution of 4-(tert-butyl)-8-(3-fluoropyridin-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (140 mg, 373 μmol, 1.0 equiv.) was stirred at 95 °C for 3 h. The reaction mixture was cooled and concentrated in vacuo to give a residue that was purified by flash column chromatography (SiO2, 0–5% MeOH in DCM) to give 4-(tert-butyl)-8-(3-fluoropyridin-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (123 mg, 44%), MS obsd. (ESI + ):375.0[(M+H) + ].

[0417] The enantiomers were separated via chiral SFC. (S)-4-(tert-butyl)-8-(3-fluoropyridin-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 59): MS obsd. (ESI + ):375.0[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d6) δ ppm: 10.41 (s, 1H), 8.78 (d, J = 2.8 Hz, 1H), 8.57 (dd, J = 5.2, 0.8 Hz, 1H), 8.08-7.99 (m, 2H), 5.36 (s, 1H), 4.82 (dd, J = 44.0, 15.2 Hz, 2H), 4.14 (d, J = 11.2 Hz, 1H), 3.46 (d, J = 11.2 Hz, 1H), 1.04 (s, 10H). (R)-4-(tert-butyl)-8-(3-fluoropyridin-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 60). MS obsd. (ESI + ):375.0[(M+H) + ].

[0418] Examples 61 and 62—Compounds 61 and 62: (S)-4-(tert-butyl)-4-hydroxy-8-(2-methylpyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 61) and (R)-4-(tert-butyl)-4-hydroxy-8-(2-methylpyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 62) (Stereochemistry for Compounds 61 and 62 is arbitrarily assigned) [ka] Step A: 10-tert-butyl-10-hydroxy-4-(2-methyl-4-pyridyl)-8,12-dioxa-5-thiatricyclo[7.4.0.0 2,6 ]Trideca-1(9),2(6),3-trien-7-one: [ka] 4-Iodo-2-methyl-pyridine was subjected to conditions similar to those described for compound 59, step F, to give 10-tert-butyl-10-hydroxy-4-(2-methyl-4-pyridyl)-8,12-dioxa-5-thiatricyclo[7.4.0.0 2,6 ] to give trideca-1(9),2(6),3-trien-7-one (169 mg, 29%). MS obsd. (ESI + ):372.2[M+H] + .

[0419] Step B: (S)-4-(tert-butyl)-4-hydroxy-8-(2-methylpyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 61) and (R)-4-(tert-butyl)-4-hydroxy-8-(2-methylpyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 62): [ka] 10-tert-butyl-10-hydroxy-4-(2-methyl-4-pyridyl)-8,12-dioxa-5-thiatricyclo[7.4.0.0 2,6 ]trideca-1(9),2(6),3-trien-7-one was subjected to conditions similar to those described for Step G of compound 59 to give 4-(tert-butyl)-4-hydroxy-8-(2-methylpyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (200 mg, 78%). MS obsd. (ESI + ):371.1[M+H] + .

[0420] The enantiomers were separated via chiral SFC. (S)-4-(tert-butyl)-4-hydroxy-8-(2-methylpyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 61): MS obsd. (ESI + ):371.0[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d6) δ 10.32 (s, 1H), 8.55 (d, J = 5.2 Hz, 1H), 7.99 (s, 1H), 7.70 (s, 1H), 7.61 (dd, J = 5.2, 1.6 Hz, 1H), 5.35 (s, 1H), 4.80 (dd, J = 38.8, 14.8 Hz, 2H), 4.14 (d, J = 11.2 Hz, 1H), 3.45 (d, J = 11.2 Hz, 1H), 2.55 (s, 3H), 1.03 (s, 9H). (R)-4-(tert-butyl)-4-hydroxy-8-(2-methylpyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 62): MS obsd. (ESI + ):371.0[(M+H) + ].

[0421] Examples 63 and 64—Compounds 63 and 64: (S)-4-(tert-butyl)-8-(5-chloropyrimidin-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 63) and (R)-4-(tert-butyl)-8-(5-chloropyrimidin-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 64) (stereochemistry for Compounds 63 and 64 is arbitrarily assigned) [ka] Step A: 5-chloro-4-iodopyrimidine: [ka] To a solution of 5-chloropyrimidin-4-amine (500 mg, 3.86 mmol, 1.0 equiv) and diiodomethane (2.07 g, 7.72 mmol, 622 μL, 2.0 equiv) in CHCN (20 mL) was added isopentyl nitrite (995 mg, 8.49 mmol, 1.14 mL, 2.2 equiv) in CHCN (1 mL) dropwise at 0 °C. The mixture was stirred at 0 °C for 30 min and then heated to 70 °C for 16 h. The reaction mixture was concentrated in vacuo, and the crude residue was purified by flash column chromatography (SiO, 0–3% EtOAc in PE) to give 5-chloro-4-iodopyrimidine (410 mg, 44% yield). MS obsd. (ESI + ):241.1[(M+H) + ].

[0422] Step B: 4-(tert-butyl)-8-(5-chloropyrimidin-4-yl)-4-hydroxy-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] Using similar conditions as described in Step F of compound 59 for (4-(tert-butyl)-4-hydroxy-6-oxo-4,6-dihydro-1H,3H-pyrano[4,3-b]thieno[3,2-d]pyran-8-yl)boronic acid and 5-chloro-4-iodopyrimidine, 4-(tert-butyl)-8-(5-chloropyrimidin-4-yl)-4-hydroxy-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (76 mg, 31%) was obtained. MS obsd. (ESI + ):393.4[(M+H) + ].

[0423] Step C: (S)-4-(tert-butyl)-8-(5-chloropyrimidin-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 63) and (R)-4-(tert-butyl)-8-(5-chloropyrimidin-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 64): [ka] Using similar conditions as described in Step G of compound 59 for 4-(tert-butyl)-8-(5-chloropyrimidin-4-yl)-4-hydroxy-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one, 4-(tert-butyl)-8-(5-chloropyrimidin-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (53 mg, 70%) was obtained as a yellow solid. MS obsd. (ESI + ):392.4[(M+H) + ].

[0424] The enantiomers were separated using chiral SFC. (S)-4-(tert-butyl)-8-(5-chloropyrimidin-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 63): MS obsd. (ESI + ):392.0[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d6) δ ppm: 10.46 (s, 1H), 9.22 (s, 1H), 9.10 (s, 1H), 8.33 (s, 1H), 5.36 (s, 1H), 4.93-4.76 (m, 2H), 4.14 (d, J = 11.2 Hz, 1H), 3.46 (d, J = 11.6 Hz, 1H), 1.04 (s, 9H). (R)-4-(tert-butyl)-8-(5-chloropyrimidin-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 64): MS obsd. (ESI + ):392.0[(M+H) + ].

[0425] Examples 65 and 66—Compounds 65 and 66: (R)-4-(tert-butyl)-8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 65) and (S)-4-(tert-butyl)-8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 66) (stereochemistry for Compounds 65 and 66 is arbitrarily assigned) [ka] Step A: 4-(tert-butyl)-8-(3-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-4-hydroxy-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] 4-Bromo-3-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole was subjected to conditions similar to those described for Step D of compound 59 to give 4-(tert-butyl)-8-(3-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-4-hydroxy-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (10 mg, 13%). MS obsd. (ESI + ):495.5[(M+H) + ].

[0426] Step B: 4-(tert-butyl)-8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] To a solution of 4-(tert-butyl)-8-(3-fluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-4-hydroxy-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (100 mg, 202 μmol, 1.0 equiv) in DCM (5 mL) was added boron trichloride (1.0 M, 1.0 mL, 5.0 equiv) at −78° C. The mixture was stirred at −78° C. for 2 h, after which MeOH was added. The mixture was concentrated in vacuo to give 4-(tert-butyl)-8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (75 mg, used without further purification). MS obsd. (ESI + ):365.5[(M+H) + ].

[0427] Step C: (R)-4-(tert-butyl)-8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 65) and (S)-4-(tert-butyl)-8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 66): [ka] 4-(Tert-butyl)-8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one was subjected to conditions similar to those described for Step G of compound 59 to give 4-(tert-butyl)-8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (40 mg, 110 μmol, 45%, 2 steps). MS obsd. (ESI + ):364.4[(M+H) + ].

[0428] The isomers were separated by chiral HPLC. (R)-4-(tert-butyl)-8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 65): MS obsd. (ESI + ):364.0[(M+H) + ]. 1H NMR (400 MHz, DMSO-d6) δ ppm: 12.83 (brs, 1H), 10.16 (brs, 1H), 8.32 (d, J = 1.8 Hz, 1H), 7.33 (s, 1H), 5.33 (s, 1H), 4.75 (dd, J = 34.2, 14.8 Hz, 2H), 4.12 (d, J = 11.2 Hz, 1H), 3.44 (d, J = 11.2 Hz, 1H), 1.03 (s, 9H). (S)-4-(tert-butyl)-8-(3-fluoro-1H-pyrazol-4-yl)-4-hydroxy-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 66). MS obsd. (ESI + ):364.0[(M+H) + ].

[0429] Examples 67 and 68—Compounds 67 and 68: (S)-4-(tert-butyl)-4-hydroxy-8-(pyridazin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 67) and (R)-4-(tert-butyl)-4-hydroxy-8-(pyridazin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 68) (Stereochemistry for Compounds 67 and 68 is arbitrarily assigned) [ka] Step A: 4-(tert-butyl)-4-hydroxy-8-(pyridazin-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] Using similar conditions as described in Step F of compound 59 for 4-bromopyridazine hydrobromide, 4-(tert-butyl)-4-hydroxy-8-(pyridazin-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (53 mg, 40%) was obtained. MS obsd. (ESI + ):359.4[(M+H) + ].

[0430] Step B: (S)-4-(tert-butyl)-4-hydroxy-8-(pyridazin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 67) and (R)-4-(tert-butyl)-4-hydroxy-8-(pyridazin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 68): [ka] Using similar conditions as described in Step G of compound 59 for 4-(tert-butyl)-4-hydroxy-8-(pyridazin-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one, 4-(tert-butyl)-4-hydroxy-8-(pyridazin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (37 mg, 66%) was obtained. MS obsd. (ESI + ):358.4[(M+H) + ].

[0431] The enantiomers were separated by chiral SFC. (S)-4-(tert-butyl)-4-hydroxy-8-(pyridazin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 67): MS obsd. (ESI + ):358.0[(M+H) + ]; 1H NMR (400 MHz, DMSO-d6) δ ppm: 10.45 (s, 1H), 9.76-9.75 (m, 1H), 9.35-9.33 (m, 1H), 8.22 (s, 1H), 8.08-8.06 (m, 1H), 5.37 (s, 1H), 4.87-4.75 (m, 2H), 4.14 (d, J = 11.6 Hz, 1H), 3.46 (d, J = 11.2 Hz, 1H), 1.03 (s, 9H). (R)-4-(tert-butyl)-4-hydroxy-8-(pyridazin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 68): MS obsd. (ESI + ):358.0[(M+H) + ].

[0432] Examples 69 and 70—Compounds 69 and 70: (S)-4-(tert-butyl)-4-hydroxy-8-(3-methylpyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 69) and (R)-4-(tert-butyl)-4-hydroxy-8-(3-methylpyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 70) (Stereochemistry for Compounds 69 and 70 is arbitrarily assigned) [ka] Step A: 4-(tert-butyl)-4-hydroxy-8-(3-methylpyridin-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one: [ka] Using similar conditions as described in Step F of compound 59 for 4-bromo-3-methyl-pyridine hydrochloride, 4-(tert-butyl)-4-hydroxy-8-(3-methylpyridin-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one (87 mg, 63%) was obtained as a yellow solid. MS obsd. (ESI + ):372.4[(M+H) + ].

[0433] Step B: (S)-4-(tert-butyl)-4-hydroxy-8-(3-methylpyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 69) and (R)-4-(tert-butyl)-4-hydroxy-8-(3-methylpyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 70): [ka] Using similar conditions as described in Step G of compound 59 for 4-(tert-butyl)-4-hydroxy-8-(3-methylpyridin-4-yl)-3,4-dihydro-1H,6H-pyrano[4,3-b]thieno[3,2-d]pyran-6-one, 4-(tert-butyl)-4-hydroxy-8-(3-methylpyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (36 mg, 41%) was obtained. MS obsd. (ESI + ):371.4[(M+H) + ].

[0434] The enantiomers were separated by chiral SFC. (S)-4-(tert-butyl)-4-hydroxy-8-(3-methylpyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 69): MS obsd. (ESI +):371.0[(M+H) + ]. 1 H NMR (400 MHz, DMSO-d₆) δ ppm: 10.30 (s, 1H), 8.60 (s, 1H), 8.52 (d, J = 4.8 Hz, 1H), 7.60 (s, 1H), 7.55 (d, J = 4.8 Hz, 1H), 5.35 (s, 1H), 4.87-4.72 (m, 2H), 4.14 (d, J = 11.2 Hz, 1H), 3.45 (d, J = 11.2 Hz, 1H), 1.04 (s, 9H). (R)-4-(tert-butyl)-4-hydroxy-8-(3-methylpyridin-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 70) (27.2 mg 29%). MS obsd.(ESI + ):371.0[(M+H) + ].

[0435] Example 71 - Compound 71: 4-methyl-8-(1H-pyrazol-4-yl)-4-(pyrrolidin-1-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one [ka] Step A: 4-hydroxy-4-methyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one: [ka] To a dry three-neck flask containing tert-butyl 4-(4-hydroxy-4-methyl-6-oxo-3,4,5,6-tetrahydro-1H-pyrano[4,3-b]thieno[3,2-d]pyridin-8-yl)-1H-pyrazole-1-carboxylate (500 mg, 1.29 mmol, 1.0 equiv.), THF (50 mL) was added, and the mixture was degassed and purged with N (3×). Methylmagnesium bromide (1 M, 25.8 mL, 20.0 equiv.) was added at 0°C and then warmed to room temperature. After 2 h, NH4Cl solution (13 mL) was added, and the mixture was extracted with EtOAc (3×50 mL). The combined organic layers were dried (MgSO), filtered, and concentrated to give a residue that was purified by flash column chromatography (SiO, 0-8% MeOH in DCM) to give 4-hydroxy-4-methyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (340 mg, 69%). MS obsd. (ESI + ):304.1[(M+H) + ].

[0436] Step B: 4-chloro-4-methyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one: [ka] Oxalyl chloride (15 mL) and N,N-dimethylformamide (0.1 mL) were added to a solution of 4-hydroxy-4-methyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (340 mg, 1.12 mmol, 1.0 equiv) in DCM (50 mL). The reaction mixture was heated to 40° C. and stirred for 16 h. The mixture was concentrated in vacuo to give 4-chloro-4-methyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (280 mg, used without further purification).

[0437] Step C: 4-methyl-8-(1H-pyrazol-4-yl)-4-(pyrrolidin-1-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 71): [ka] A solution of 4-chloro-4-methyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (280 mg, crude) in acetonitrile (40 mL) was cooled to 0° C., and then pyrrolidine (15 mL) was added dropwise. The reaction mixture was allowed to warm to room temperature for 16 hours, then concentrated and purified by reverse-phase column chromatography (0-20% MeCN in water (0.1% FA)). Further purification by preparative TLC (SiO, 0-10% MeOH in DCM) gave 4-methyl-8-(1H-pyrazol-4-yl)-4-(pyrrolidin-1-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 71) (2.2 mg, 17%). MS obsd. (ESI + ):357.2[(M+H) + ]. 1 H NMR(400MHz,DMSO-d6)δ ppm:8.11(s,2H),7.40(s,1H),4.74(d,J=14.2Hz,1H),4.61(d,J=14.2Hz,1H),4.12(d,J=11.6Hz,1H),3.5 8(d,J=11.8Hz,1H),2.80(d,J=6.4Hz,2H),2.44-2.38(m,1H),2.06-1.92(m,1H),1.65(s,4H),1.42(s,3H).

[0438] Examples 72 and 73—Compounds 72 and 73: (S)-4-hydroxy-4-(prop-1-yn-1-yl)-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 72) and (R)-4-hydroxy-4-(prop-1-yn-1-yl)-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 73) (stereochemistry for Compounds 72 and 73 is arbitrarily assigned) [ka] tert-Butyl 4-(7,10-dioxo-12-oxa-5-thia-8-azatricyclo[7.4.0.0]) in 1,4-dioxane (20 mL) 2,6 To a solution of ]trideca-1(9),2(6),3-trien-4-yl)pyrazole-1-carboxylate (100 mg, 258 μmol, 1.0 equiv.) was added prop-1-yn-1-ylmagnesium bromide (0.5 M, 10 mL, 20 equiv.) at 0° C. The mixture was stirred at 0° C. for 1 h, and then HO (20 mL) was added slowly at 0° C. The resulting mixture was extracted with EtOAc (3×350 mL), and the combined organic phases were dried (NaSO), filtered, and concentrated in vacuo. The crude product was purified by flash silica gel column chromatography (SiO, 0-10% MeOH in DCM) to give 4-hydroxy-4-(prop-1-yn-1-yl)-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (110 mg, 43%). MS obsd. (ESI + ):328.0[(M+H) + ].

[0439] The enantiomers were separated by chiral SFC. (S)-4-Hydroxy-4-(prop-1-yn-1-yl)-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 72): MS obsd. (ESI + ):328.1[(M+H) + ]; 1 H NMR (400 MHz, DMSO-d6) δ ppm: 13.23 (s, 1H), 10.90 (s, 1H), 8.29 (s, 1H), 7.97 (s, 1H), 7.43 (s, 1H), 6.20 (s, 1H), 4.74 (s, 2H), 3.94 (d, J = 11.2 Hz, 1H), 3.76 (d, J = 11.2 Hz, 1H), 1.86 (s, 3H). (R)-4-Hydroxy-4-(prop-1-yn-1-yl)-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 73). MS obsd. (ESI + ):327.8[(M+H) + ].

[0440] Examples 74, 75, 76, and 77—Compounds 74, 75, 76, and 77: (S)-4-((S)-sec-butyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 74), (R)-4-((R)-sec-butyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 75), (S )-4-((R)-sec-butyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 76), and (R)-4-((S)-sec-butyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 77) (stereochemistry for Compounds 74, 75, 76, and 77 is arbitrarily assigned). [ka] A procedure similar to compound 72, step A, was carried out using tert-pentylmagnesium chloride (1 M in THF, 7.74 mL, 30 equivalents) to give 4-(sec-butyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (35.2 mg, 39%). MS obsd. (ESI + ):346.2[(M+H) + ].

[0441] The four diastereomers were separated by chiral SFC. (S)-4-((S)-sec-butyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 74): MS obsd. (ESI + ):346.2[(M+H) + ].1 H NMR(400MHz,DMSO-d6)δ ppm:13.22(s,1H),10.80(s,1H),8.28(brs,1H),7.96(brs,1H),7.42(s,1H),5.27(s,1H),4.77 -4.53(m,2H),3.92(d,J=11.6Hz,1H),3.56(d,J=11.6Hz,1H),2.12-2.07(m,1H),1.87-1.80(m, 1H), 1.09-0.97 (m, 1H), 0.92 (t, J = 7.2 Hz, 3H), 0.72 (d, J = 7.2 Hz, 3H). (R)-4-((R)-sec-butyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 75): MS obsd. (ESI + ):346.0[(M+H) + ]. 1 H NMR(400MHz,DMSO-d6)δ ppm:13.22(s,1H),10.82(s,1H),8.28(brs,1H),7.96(brs,1H),7.41(s,1H),5.24(s,1H), 4.73-4.53(m,2H),3.93(d,J=11.6Hz,1H),3.54(d,J=11.6Hz,1H),2.13-2.02(m,1H),1.15 -1.05 (m, 2H), 0.98 (d, J = 7.2 Hz, 3H), 0.77 (t, J = 7.2 Hz, 3H). (S)-4-((R)-sec-butyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 76). MS obsd. (ESI + ):346.0[(M+H) + ]. 1H NMR(400MHz,DMSO-d6)δ ppm:13.22(s,1H),10.82(s,1H),8.09(brs,2H),7.41(s,1H),5.25(s,1H),4.72-4.62( m,2H),3.93(d,J=11.6Hz,1H),3.54(d,J=11.6Hz,1H),2.13-2.02(m,1H),1.13-1.09(m , 2H), 0.98 (d, J = 7.2 Hz, 3H), 0.76 (d, J = 7.2 Hz, 3H). (R)-4-((S)-sec-butyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 77): MS obsd. (ESI + ):346.0[(M+H) + ]. 1 H NMR(400MHz,DMSO-d6)δ ppm:13.22(s,1H),10.81(s,1H),8.27(brs,1H),7.96(brs,1H),7.42(s,1H),5.27(s,1H),4.65(t,J=9.2Hz,2H),3.92(d,J=11.6Hz) ,1H),3.56(d,J=11.6Hz,1H),2.14-2.03(m,1H),1.91-1.82(m,1H),0.97-1.10(m,1H),0.92(t,J=7.2Hz,3H),0.72(d,J=7.2Hz,3H).

[0442] Examples 78 and 79 - Compounds 78 and 79: (S)-4-cyclopentyl-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 78) and (R)-4-cyclopentyl-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 79) (stereochemistry for Compounds 78 and 79 is arbitrarily assigned) [ka] Cyclopentylmagnesium bromide (17 mmol, 17 mL, 10 equiv.) was added rapidly to a solution of tert-butyl 4-(4,6-dioxo-3,4,5,6-tetrahydro-1H-pyrano[4,3-b]thieno[3,2-d]pyridin-8-yl)-1H-pyrazole-1-carboxylate (1.0 g, 1.73 mmol, 1.0 equiv.) in THF (100 mL) at room temperature. The reaction mixture was degassed, purged with N2, and stirred at room temperature for 2 h. NH4Cl solution (160 mL) was added, and the mixture was extracted with EtOAc (3 x 40 mL). The combined organic layers were dried (Na2SO4), filtered, and concentrated. Column chromatography (SiO2, 0-5% MeOH in DCM) followed by reverse-phase column chromatography (C18-SiO2, 0-20% MeCN in water (0.1% FA)) gave 4-cyclopentyl-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (130 mg, 14%). MS obsd. (ESI + ):358.1[(M+H) + ].

[0443] The enantiomers were separated by chiral SFC. (S)-4-Cyclopentyl-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 78): MS obsd. (ESI + ):357.8[(M+H) + ]. 1H NMR (400 MHz, DMSO-d6) δ ppm: 13.23 (s, 1H), 10.62 (s, 1H), 8.11 (brs, 2H), 7.40 (s, 1H), 5.29 (s, 1H), 4.71 (q, J = 14.4 Hz, 2H), 3.90 (d, J = 11.2 Hz, 1H), 3.57 (d, J = 11.2 Hz, 1H), 1.69-1.37 (m, 9H). (R)-4-Cyclopentyl-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 79). MS obsd. (ESI + ):357.8[(M+H) + ].

[0444] Examples 80 and 81 - Compounds 80 and 81: (S)-4-hydroxy-4-(tert-pentyl)-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 80) and (R)-4-hydroxy-4-(tert-pentyl)-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 81) (Stereochemistry for Compounds 80 and 81 is arbitrarily assigned) [ka] A procedure similar to Step A of compound 72 was carried out using chloro(1,1-dimethylpropyl)magnesium to give 4-(sec-butyl)-4-hydroxy-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (30 mg, 32%). MS obsd. (ESI + ):360.2[(M+H) + ].

[0445] The enantiomers were separated by chiral SFC. (S)-4-Hydroxy-4-(tert-pentyl)-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 80): MS obsd. (ESI + ):360.2[(M+H) + ]; 1 H NMR(400MHz,DMSO)δ 13.23(s,1H),9.99(s,1H),8.29(brs,1H),7.96(brs,1H),7.40(s,1H),5.31(s,1H),4. 74(q,J=14.8Hz,2H),4.17(d,J=11.4Hz,1H),3.44(d,J=11.4Hz,1H),1.63-1.56(m,1H), 1.33-1.38 (m, 1H), 0.96 (d, J = 6.0 Hz, 7H), 0.76 (s, 2H). (R)-4-Hydroxy-4-(tert-pentyl)-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (compound 81): MS obsd. (ESI + ):360.2[(M+H) + ].

[0446] Examples 82 and 83—Compounds 82 and 83: (S)-4-(dimethylamino)-4-methyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 82) and (R)-4-(dimethylamino)-4-methyl-8-(1H-pyrazol-4-yl)-1,3,4,5-tetrahydro-6H-pyrano[4,3-b]thieno[3,2-d]pyridin-6-one (Compound 83) (stereochemistry for Compounds 82 and 83 is arbitrarily assigned) [ka] Step A: N-(4,4-dimeth...

Claims

1. Compounds of formula (I) 【Chemistry 1】 or a pharmaceutically acceptable salt thereof, During the ceremony, R １ is hydrogen or halogen, R ２ is a 5- to 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from the group consisting of halogen and C1-C6 alkyl, or is a 5- to 6-membered heterocyclyl optionally substituted with 1 or 2 substituents independently selected from the group consisting of halogen and C1-C6 alkyl; R ３ is hydrogen or C1-C6 alkyl; Ring A is C6-C10 cycloalkyl or 6- to 10-membered heterocyclyl; Each R ４ is halogen, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 hydroxyalkyl, C1-C6 haloalkyl, C1-C6 alkoxy(C1-C6 alkyl)-, —C(═O)C1-C6 alkyl, C2-C6 alkynyl, C3-C6 cycloalkyl, —NR Ａ R Ｂ and 4- to 6-membered heterocyclyl optionally substituted with 1 or 2 independently selected halogens; Each R Ａ and R Ｂ are independently hydrogen or C1-C6 alkyl; A compound of formula (I), or a pharmaceutically acceptable salt thereof, wherein m is 0, 1, 2, 3, or 4.

2. 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein m is 1, 2, 3, or 4.

3. R ２ is a 5-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from the group consisting of halogen and C1-C6 alkyl, or a pharmaceutically acceptable salt thereof.

4. R ２ 2. The compound of claim 1, wherein said 5-membered heteroaryl is pyrazolyl or isothiazolyl, or a pharmaceutically acceptable salt thereof.

5. R ２ 2. The compound of claim 1, wherein said 5-membered heteroaryl is 4-pyrazolyl, 5-pyrazolyl, or 5-isothiazolyl, or a pharmaceutically acceptable salt thereof.

6. R ２ is a 6-membered heteroaryl optionally substituted with 1 or 2 substituents independently selected from the group consisting of halogen and C1-C6 alkyl, or a pharmaceutically acceptable salt thereof.

7. R ２ 2. The compound of claim 1, wherein said 6-membered heteroaryl is pyridyl, pyrimidinyl, or pyridazinyl, or a pharmaceutically acceptable salt thereof.

8. R ２ 2. The compound of claim 1, wherein said 6-membered heteroaryl is 4-pyridyl, 4-pyrimidinyl, or 4-pyridazinyl, or a pharmaceutically acceptable salt thereof.

9. R ２ 2. The compound of claim 1, wherein said heteroaryl is substituted with one substituent selected from halogen and C1-C6 alkyl, or a pharmaceutically acceptable salt thereof.

10. R ２ 2. The compound of claim 1, wherein said heteroaryl is substituted with one substituent selected from fluoro, chloro, and methyl, or a pharmaceutically acceptable salt thereof.

11. R ２ 2. The compound of claim 1, wherein said heteroaryl is substituted with two substituents independently selected from halogen and C1-C6 alkyl, or a pharmaceutically acceptable salt thereof.

12. R ２ 2. The compound of claim 1, wherein said heteroaryl is substituted with two substituents independently selected from fluoro and methyl, or a pharmaceutically acceptable salt thereof.

13. R ２ The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is unsubstituted 5-6 membered heteroaryl.

14. R ２ 14. The compound of claim 13, or a pharmaceutically acceptable salt thereof, wherein is selected from the group consisting of pyrazolyl, isothiazolyl, pyridyl, and pyridazinyl.

15. R ２ The compound of claim 13, or a pharmaceutically acceptable salt thereof, wherein is 4-pyrazolyl, 5-isothiazolyl, 4-pyridyl, or 4-pyridazinyl.

16. R ２ is a 5- to 6-membered heterocyclyl optionally substituted with 1 or 2 substituents independently selected from the group consisting of halogen and C1-C6 alkyl, or a pharmaceutically acceptable salt thereof.

17. R ２ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is morpholinyl.

18. R ２ The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is 4-morpholinyl.

19. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein ring A is C6-C10 cycloalkyl.

20. 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein ring A is cyclohexyl or bicyclo[2.2.2]octanyl.

21. 2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein ring A is a 6- to 10-membered heterocyclyl.

22. 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein ring A is tetrahydropyranyl or piperidinyl.

23. One R ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is halogen.

24. One R ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is fluoro.

25. One R ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is hydroxyl.

26. One R ４ The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is C1-C6 alkyl.

27. One R ４ But -CH ３ , -CH ２ CH ３ , -CH(CH ３ ) ２ , -CH(CH ３ ) CH ２ CH ３ , -C(CH ３ ) ２ CH ２ CH ３ , and -C(CH ３ ) ３ 10. The compound of claim 1, selected from the group consisting of:

28. One R ４ The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein is C1-C6 alkoxy.

29. One R ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is methoxy.

30. One R ４ The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein is C1-C6 hydroxyalkyl.

31. One R ４ But -CH ２ OH or -C(CH ３ ) ２ 10. The compound of claim 1, wherein R is OH, or a pharmaceutically acceptable salt thereof.

32. One R ４ The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein is C1-C6 haloalkyl.

33. One R ４ But, -CF ３ , -CHF 2 , -CF 2 CH 3 , or -CH 2 CF 2 CH 3, or a pharmaceutically acceptable salt thereof.

34. One R ４ The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein is C1-C6 alkoxy(C1-C6 alkyl)-.

35. One R ４ But -CH ２ OCH ３ 2. The compound of claim 1, wherein:

36. R ４ or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, wherein one of

37. One R ４ is -C(=O)CH ３ 2. The compound of claim 1, wherein:

38. One R ４ is C2-C6 alkynyl, or a pharmaceutically acceptable salt thereof.

39. One R ４ The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is 1-propynyl.

40. One R ４ is C3-C6 cycloalkyl, or a pharmaceutically acceptable salt thereof.

41. One R ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: is cyclopentyl.

42. One R ４ But, -NR Ａ R Ｂ and one of R A and R B is hydrogen and the other of R A and R B is C1-C6 alkyl; R A and R B are both C1-C6 alkyl; or R A and R B are both H; 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof.

43. One R ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is 4-6 membered heterocyclyl optionally substituted with one or two independently selected halogens.

44. One R ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: is pyrrolidinyl or azetidinyl, wherein said pyrrolidinyl or azetidinyl is optionally substituted with one or two independently selected halogens.

45. One R ４ The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein is 1-pyrrolidinyl or 1-azetidinyl.

46. m is 2, 3, or 4, and two R ４ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the group is geminal.

47. m is 2, and two R ４ The base is geminal, On the other hand, R ４ is hydroxyl, and the other R ４ is C1-C6 alkyl; each R 4 is fluoro; one R 4 is hydroxyl and the other R 4 is C1-C6 haloalkyl; one R 4 is hydroxyl and the other R 4 is C2-C6 alkynyl; one R 4 is hydroxyl and the other R 4 is C3-C6 cycloalkyl; one R 4 is hydroxyl and the other R 4 is C1-C6 alkoxy(C1-C6 alkyl)-; each R 4 is an independently selected C1-C6 alkyl; one R 4 is C1-C6 alkyl and the other R 4 is C1-C6 hydroxyalkyl; one R 4 is C1-C6 alkyl and the other R 4 is C1-C6 alkoxy; one R 4 is C1-C6 alkyl and the other R 4 is —NR A R B , or one R 4 is C1-C6 alkyl and the other R 4 is 4-6 membered heterocyclyl optionally substituted with 1 or 2 independently selected halogens; 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof.

48. m is 4 and two R ４ The base is geminal, Two R's ４ is fluoro and one R ４ is C1-C6 alkoxy, and one R ４ is C1-C6 alkyl; two R 4 are fluoro, one R 4 is C1-C6 hydroxyalkyl, and one R 4 is C1-C6 alkyl; two R 4 are fluoro, one R 4 is hydroxyl, and one R 4 is C1-C6 alkyl; or two R 4 are fluoro, one R 4 is hydroxyl, and one R 4 is C1-C6 alkoxy(C1-C6 alkyl)-; 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof.

49. 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein m is 0.

50. R １ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is halogen.

51. R １ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is fluoro.

52. R １ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is hydrogen.

53. R ３ 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is hydrogen.

54. R ３ The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein is C1-C6 alkyl.

55. The compound of formula (I) is a compound of formula (IA): 【Chemistry 2】 2. The compound of claim 1, wherein:

56. The compound of formula (I) is a compound of formula (IB): 【Transformation 3】 2. The compound of claim 1, wherein:

57. The compound of formula (I) is a compound of formula (IC): 【Chemistry 4】 2. The compound of claim 1, wherein:

58. The compound of formula (I) is a compound of formula (ID): 【Transformation 5】 2. The compound of claim 1, wherein:

59. The compound of formula (I) is a compound of formula (IP): 【Chemistry 17】 2. The compound of claim 1, wherein:

60. The compound is 【Chemistry 18-1】 【Chemistry 18-2】 【Chemistry 18-3】 【Chemistry 18-4】 【Chemistry 18-5】 【Chemistry 18-6】 【Chemistry 18-7】 【Chemistry 18-8】 10. The compound of claim 1, selected from the group consisting of:

61. A compound, or a pharmaceutically acceptable salt thereof, comprising: 【Chemistry 19】 or a pharmaceutically acceptable salt thereof.

62. A compound, or a pharmaceutically acceptable salt thereof, comprising: 【Chemistry 20】 or a pharmaceutically acceptable salt thereof.

63. A compound, or a pharmaceutically acceptable salt thereof, comprising: 【Chemistry 21】 or a pharmaceutically acceptable salt thereof.

64. A compound, or a pharmaceutically acceptable salt thereof, comprising: 【Chemistry 22】 or a pharmaceutically acceptable salt thereof.

65. A compound, or a pharmaceutically acceptable salt thereof, comprising: 【Chemistry 23】 or a pharmaceutically acceptable salt thereof.

66. A compound, or a pharmaceutically acceptable salt thereof, comprising: 【Chemistry 24】 or a pharmaceutically acceptable salt thereof.

67. A compound, or a pharmaceutically acceptable salt thereof, comprising: 【Chemistry 25】 or a pharmaceutically acceptable salt thereof.

68. A compound, or a pharmaceutically acceptable salt thereof, comprising: 【Chemistry 26】 or a pharmaceutically acceptable salt thereof.

69. A compound, or a pharmaceutically acceptable salt thereof, comprising: 【Chemistry 27】 or a pharmaceutically acceptable salt thereof.

70. A compound, or a pharmaceutically acceptable salt thereof, comprising: 【Chemistry 28】 or a pharmaceutically acceptable salt thereof.

71. A compound, or a pharmaceutically acceptable salt thereof, comprising: 【Chemistry 29】 or a pharmaceutically acceptable salt thereof.

72. A compound, or a pharmaceutically acceptable salt thereof, comprising: 【Transformation 30】 or a pharmaceutically acceptable salt thereof.

73. 73. A pharmaceutical composition comprising a compound according to any one of claims 1 to 72, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

74. 73. A composition for treating cancer in a subject in need thereof, comprising a compound of any one of claims 1 to 72, or a pharmaceutically acceptable salt thereof.

75. The cancer is acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myelomonocytic leukemia (CMML), myelodysplastic syndrome (MDS), myeloproliferative disorder (MPD), bladder cancer, breast cancer, cervical cancer, colon cancer, colorectal cancer, endometrial cancer, esophageal cancer, Ewing's sarcoma, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), head and neck cancer, head and neck neoplasm, hepatocellular carcinoma, pancreatic neuroendocrine tumor, bone cancer, melanoma, non-small cell lung cancer, lung neoplasm, lung cancer, pulmonary neoplasm neoplasms), osteosarcoma, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, squamous cell cervical cancer, gastric cancer, uterine cancer, or uterine sarcoma.

76. The composition described in claim 74, wherein the cancer is myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), or acute lymphocytic leukemia (ALL).

77. The composition described in claim 74, wherein the cancer is prostate cancer or small cell lung cancer.

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