Cdk inhibitors

Abstract

To provide Cyclin-Dependent Kinase inhibitors which can be used in the treatment of cell proliferative disorders such as cancer.SOLUTION: Provided is a compound represented by structural formula (I), or a pharmaceutically acceptable salt, or a stereoisomer thereof. In the formula, ring A is a specific 5- or 6-membered condensed heteroaryl; ring B is a bond, a 3- to 10-membered heterocyclyl, or a 5- to 10-membered heteroaryl; ring C is a 5- to 6-membered heteroaryl, a 5- to 10-membered heterocyclyl, a phenyl, or a 5- to 10-membered bridged bicyclic group; L is a linker; each R1 is H, deuterium, halogen, -OH, C1-4 alkyl, or the like; each occurrence of R2 is H, deuterium, halogen, -OH, CN, C1-8 alkyl, or the like.SELECTED DRAWING: None

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims the benefit of priority to International Patent Application No. PCT / CN2019 / 085494, filed on May 5, 2019. The entire content of the foregoing application is incorporated herein by reference.

Background Art

[0002] Cyclin - dependent kinases (CDKs) are a family of protein kinases first discovered for their role in regulating the cell cycle. Since then, they have been confirmed to play roles in regulating many other biological functions, such as transcription, mRNA processing, and neuronal differentiation.

[0003] CDKs are relatively small proteins with a molecular weight of approximately 34 - 40 kDa. They contain almost only the kinase domain and are essentially inactive when not forming a complex with a class of regulatory proteins called cyclins. The levels of CDKs remain relatively constant throughout the cell cycle, and most regulation is post - translational, most notably by binding to cyclins.

[0004] Similar to all kinases, the active site or ATP-binding site of CDK is a cleft between a small amino-terminal lobe and a large carboxy-terminal lobe. From the structure of human CDK2, it has become clear that CDK has a modified ATP-binding site that can be regulated by cyclin binding. Phosphorylation by CDK-activating kinase (CAK) at Thr161 on the T-loop increases complex activity. In the absence of cyclin, a flexible loop called the activation loop or T-loop closes the cleft, and the positions of some important amino acid residues become suboptimal for ATP binding. In the presence of cyclin, two α-helices change position to enable ATP binding. One of them, the L12 helix immediately preceding the T-loop in the primary sequence, becomes a β-strand and helps to rearrange the T-loop so that the T-loop no longer blocks the active site. Another α-helix, called the PSTAIRE helix, rearranges and helps to change the positions of important amino acid residues at the active site.

[0005] Therefore, only cyclin-CDK complexes have active kinase activity, and most known cyclin-CDK complexes regulate progression through the cell cycle. CDKs are ubiquitous in all known eukaryotes, and their regulatory functions in the cell cycle are evolutionarily conserved. For example, yeast cells can grow normally when their CDK genes are replaced with homologous human genes. CDKs exert their regulatory functions by phosphorylating certain serine and threonine residues, as well as substrates on the consensus sequence of [S / T]PX[K / R] (where S / T is the target Ser or Thr for phosphorylation, P is proline, X is any amino acid, K is lysine, and R is arginine).

[0006] Animal cells have at least nine different CDKs, four of which (CDK1, 2, 3, and 4) are directly involved in the regulation of the cell cycle. In mammalian cells, only CDK1 and its binding partners cyclin A2 and B1 can drive the cell cycle. Cyclin-CDK complexes in the early cell cycle stages help activate cyclin-CDK complexes in later stages.

[0007] The same CDK can form complexes with different cyclins to regulate different stages of the cell cycle. For example, CDK2 can form a complex with cyclin D or E to regulate the G1 phase, with cyclin A or E to regulate the S phase, and with cyclin A to regulate the G2 phase. On the other hand, CDK4 and CDK6 can form complexes with cyclin D1, D2, and D3.

[0008] Highly homologous cyclin-dependent kinases (CDKs) CDK4 and CDK6 in combination with cyclin D are important regulators of the transition through the restriction point R between the G1 (growth) and S (DNA replication) phases of the cell cycle. CDK4 / 6 exerts its effect through the phosphorylation of the retinoblastoma protein (pRb). Once phosphorylated, pRb loses its inhibitory effect on the transcription of genes that promote the transition to the S phase.

[0009] On the other hand, specific inhibition of CDK4 / 6 kinase activity by the endogenous protein modulator p16 INK4 or small molecule inhibitors results in hypophosphorylated pRb and cell arrest at the G1 restriction point. As a major mechanism for regulating the G1 restriction point, the pathways regulated by these kinases vary in a wide range of human tumors, and thus, inhibition of CDK4 / CDK6 in these tumors has therapeutic utility by preventing cell division.

[0010] There is still a need to provide CDK4 / 6 inhibitors that can be used in the treatment of cell proliferative diseases such as cancer.

Summary of the Invention

[0011] Described herein are compounds of formula (I), (II-A)-(II-J), and the compounds of the Examples (collectively referred to herein as "the compounds of the invention"), and pharmaceutically acceptable salts or stereoisomers thereof, which inhibit the activity of cyclin-dependent kinases (CDKs), such as CDK2, CDK4, and / or CDK6.

[0012] In one aspect, the invention provides a compound of formula (I), or a pharmaceutically acceptable salt or stereoisomer thereof,

Chemical formula

[0013] In one embodiment, the compound, or a pharmaceutically acceptable salt or stereoisomer thereof, is selected from the compounds of the Examples provided herein.

[0014] Also provided is a pharmaceutical composition comprising a compound of the invention, or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier.

[0015] The present disclosure further provides a method for treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of (1) a compound of the invention, or a pharmaceutically acceptable salt or stereoisomer thereof, or (2) a pharmaceutically acceptable composition comprising a compound of the invention, or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier. In certain embodiments, the cancer is selected from the group consisting of colorectal cancer, breast cancer (such as hormone receptor-positive, HER2 / neu-negative advanced or metastatic breast cancer in postmenopausal women), lung cancer, prostate cancer, glioblastoma, mantle cell lymphoma, chronic myeloid leukemia, and acute myeloid leukemia. ​

[0016] In certain embodiments of the method of the present invention, cancer can be treated by inhibiting the activity of cyclin-dependent kinases (CDKs), such as CDK2, CDK4, and / or CDK6.

[0017] In certain embodiments of the method of the present invention, the cancer is bladder, breast, colon, kidney, epidermal, liver, lung, esophageal, gallbladder, ovarian, pancreatic, stomach, cervical, thyroid, nasal, head and neck, prostate, or skin cancer, lymphatic hematopoietic tumors, myeloid hematopoietic tumors, thyroid follicular cancer, tumors of mesenchymal origin, tumors of the central or peripheral nervous system, melanoma, seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum, corneal tumors, thyroid follicular cancer, or Kaposi's sarcoma.

[0018] In certain embodiments of the method of the present invention, the compounds of the present invention are administered together with any one of the second therapeutic agents described herein that also treat cancer.

[0019] The present disclosure also provides the use of a compound of the present invention, or a pharmaceutically acceptable salt or stereoisomer thereof, or a pharmaceutical composition comprising the same, in any of the methods of the present invention described above. In one embodiment, there is provided a compound of the present invention, or a pharmaceutically acceptable salt or stereoisomer thereof, or a pharmaceutical composition comprising the same, for use in any of the methods of the present invention described above. In another embodiment, there is provided the use of a compound of the present invention or a pharmaceutically acceptable salt or stereoisomer thereof, or a pharmaceutical composition comprising the same, for the manufacture of a medicament for any of the methods of the present invention described.

DETAILED DESCRIPTION OF THE INVENTION

[0020] 1. Overview The present invention provides a compound of the present invention or a pharmaceutically acceptable salt thereof for use in treatments such as cancer treatment.

[0021] The present invention also provides a pharmaceutical formulation comprising a compound of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, or excipient.

[0022] The present invention provides a compound of the present invention or a pharmaceutically acceptable salt thereof for use in the treatment of cancer. In particular, these cancers can be any of the cancers described below, for example, colorectal cancer, breast cancer (including ER + HER2 - progressive or metastatic or recurrent breast cancer in adult or postmenopausal women), lung cancer, particularly non-small cell lung cancer (NSCLC), prostate cancer, glioblastoma, mantle cell lymphoma (MCL), chronic myeloid leukemia (CML), and acute myeloid leukemia (AML).

[0023] The present invention further provides a method for treating a cancer selected from the group consisting of colorectal cancer, breast cancer (including ER + HER2 - progressive or metastatic or recurrent breast cancer in adult or postmenopausal women), lung cancer, particularly non-small cell lung cancer (NSCLC), prostate cancer, glioblastoma, mantle cell lymphoma (MCL), chronic myeloid leukemia, and acute myeloid leukemia in a mammal, the method comprising administering to a mammal in need of such treatment an effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof.

[0024] Furthermore, the present invention provides the use of a compound of the present invention or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of cancer. In particular, these cancers are selected from the group consisting of colorectal cancer, breast cancer (including ER progressive or metastatic or recurrent breast cancer in adult or postmenopausal women), lung cancer, particularly non-small cell lung cancer (NSCLC), prostate cancer, glioblastoma, mantle cell lymphoma (MCL), chronic myeloid leukemia, and acute myeloid leukemia. + HER2 -

[0025] ​Furthermore, the present invention provides a pharmaceutical preparation for therapeutic use, comprising a compound of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, or excipient. The present invention also provides a pharmaceutical preparation for treating colorectal cancer, breast cancer (including in adult or postmenopausal women with ER + HER2 - advanced or metastatic or recurrent breast cancer), lung cancer, particularly non-small cell lung cancer (NSCLC), prostate cancer, glioblastoma, mantle cell lymphoma (MCL), chronic myeloid leukemia and acute myeloid leukemia, comprising a compound of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, or excipient.

[0026] The indications of treatable diseases and potential second therapeutic agents useful in combination therapies are described in more detail in the following section.

[0027] It should be understood that any one or more additional embodiments of the present invention can be combined with any of the embodiments described herein, including those described in only one of the following sections or only in the examples, unless explicitly denied or otherwise inappropriate / inapplicable.

[0028] 2. Definitions As used herein, the terms "halo" or "halogen" mean halogen and include chloro, fluoro, bromo, and iodo.

[0029] The term "alkyl", used alone or as part of a larger moiety such as "alkoxy" or "haloalkyl", means a saturated aliphatic straight or branched chain monovalent hydrocarbon radical. Unless otherwise specified, alkyl groups typically have 1 to 4 carbon atoms, i.e., are (C1-C4) alkyl. As used herein, the "(C1-C4) alkyl" group means a radical having 1 to 4 carbon atoms in a straight or branched chain arrangement. Examples include methyl, ethyl, Examples include n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl.

[0030] The term "alkenyl" means a branched or straight-chain monovalent hydrocarbon radical containing at least one double bond. Alkenyl can be mono- or poly-unsaturated and can exist in the E or Z configuration. Unless otherwise specified, an alkenyl group typically has 2 to 6 carbon atoms, i.e., (C2-C6) alkenyl. For example, "(C2-C6) alkenyl" means a radical having 2 to 6 carbon atoms in a straight-chain or branched-chain arrangement.

[0031] The term "alkynyl" means a branched or straight-chain monovalent hydrocarbon radical containing at least one triple bond. Unless otherwise specified, an alkynyl group typically has 2 to 6 carbon atoms, i.e., (C2-C6) alkynyl. For example, "(C2-C6) alkynyl" means a radical having 2 to 6 carbon atoms in a straight-chain or branched-chain arrangement.

[0032] The term "alkoxy" means an alkyl radical bonded through an oxygen-bonded atom, represented by -O-alkyl. For example, "(C1-C4) alkoxy" includes methoxy, ethoxy, propoxy, and butoxy.

[0033] The terms "haloalkyl" and "haloalkoxy" mean alkyl or alkoxy optionally substituted with one or more halogen atoms. Examples of haloalkyl include, but are not limited to, trifluoromethyl, trichloromethyl, pentafluoroethyl, etc.

[0034] The terms "hydroxyalkyl" and "hydroxyalkoxy" mean alkyl or alkoxy optionally substituted with one or more hydroxy groups.

[0035] As used herein, the term "cycloalkyl" refers to a saturated cyclic, bicyclic, tricyclic, or polycyclic hydrocarbon group having 3 to 14 carbon atoms including the indicated number of rings and carbon atoms (e.g., C3-C 14 monocyclic, C4-C 14 bicyclic, C5-C 14 tricyclic, or C6-C 14 polycyclic cycloalkyl). In some embodiments, "cycloalkyl" is monocyclic cycloalkyl. Examples of monocyclic cycloalkyl groups include cyclopentyl (C5), cyclohexyl (C5), cyclopropyl (C3), cyclobutyl (C4), cycloheptyl (C7), and cyclooctyl (C8). In some embodiments, "cycloalkyl" is bicyclic cycloalkyl. Examples of bicyclic cycloalkyl include bicyclo[1.1.0]butane (C4), bicyclo[1.1.1]pentane (C5), spiro[2.2]pentane (C5), bicyclo[2.1.0]pentane (C5), bicyclo[2.1.1]hexane (C6), bicyclo[3.3.3]undecane (C 11 ), decahydronaphthalene (C 10 ), bicyclo[4.3.2]undecane (C 11 ), spiro[5.5]undecane (C 11 ), and bicyclo[4.3.3]dodecane (C 12 ). In some embodiments, "cycloalkyl" is tricyclic cycloalkyl. Examples of tricyclic cycloalkyl include adamantane (C 12 ). Unless otherwise indicated, "cycloalkyl" has 3 to 6 carbon atoms.

[0036] The term "aryl group", when used alone or as part of a larger moiety such as in the case of "aralkyl", "aralkoxy", or "aryloxyalkyl", means a carbocyclic aromatic ring. The term "aryl" may be used interchangeably with the terms "aryl ring", "carbocyclic aromatic ring", "aryl group", and "carbocyclic aromatic group". An aryl group typically has 6 to 14 ring atoms. Examples include phenyl, naphthyl, anthracenyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, fluorenyl, indanyl, indenyl, etc. A "substituted aryl group" is substituted at any one or more of the ring atoms that are ring carbon atoms bonded to hydrogen.

[0037] The term "heterocyclyl group" or "heterocyclic group" means a 3- to 10-membered monocyclic non-aromatic ring having 1 to 4 ring heteroatoms, or a polycyclic ring having a 7- to 20-membered ring and 1 to 4 ring heteroatoms, where the polycyclic ring has one or more monocyclic non-aromatic heterocyclic rings fused to one or more aromatic or heteroaromatic rings. Each heteroatom is independently selected from nitrogen, quaternary nitrogen, nitrogen oxide (e.g., NO), air, and sulfur including sulfoxide and sulfone. In one embodiment, the heterocyclyl group is a bicyclic ring having a monocyclic non-aromatic heterocyclic ring fused to a phenyl group. Exemplary polycyclic heterocyclic groups include tetrahydroisoquinolinyl (1,2,3,4-tetrahydroisoquinolin-7-yl, 2-methyl-1,2,3,4-tetrahydroisoquinolin-7-yl, 1,2,3,4-tetrahydroisoquinolin-6-yl, and 2-methyl-1,2,3,4-tetrahydroisoquinolin-6-yl, etc.), isoindolinyl (2-ethylisoind Examples include 5 - pyridyl, 2 - methylisoindolin - 5 - yl, etc., indolinyl, tetrahydrobenzo[f]oxazepine (such as 2,3,4,5 - tetrahydrobenzo[f][1,4]oxazepin - 7 - yl). The terms "heterocyclic", "heterocyclyl", or "heterocyclic ring" also refer to rings that may be saturated or partially unsaturated and are optionally substituted. In some embodiments, the heterocyclyl group is a 3 - to 14 - membered non - aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, each heteroatom independently selected from nitrogen, oxygen, and sulfur ("3 - to 14 - membered heterocyclyl").

[0038] The terms "heteroaryl", "heteroaromatic", "heteroaryl ring", "heteroaryl group", "heteroaromatic ring", and "heteroaromatic group", used alone or as part of a larger moiety as in the case of "heteroalkyl" or "heteroarylalkoxy", refer to aromatic ring groups having 5 to 14 ring atoms selected from carbon and at least one (typically 1 to 4, more typically 1 or 2) heteroatoms (e.g., oxygen, nitrogen, or sulfur). "Heteroaryl" includes monocyclic and polycyclic rings, where a monocyclic heteroaromatic ring is fused to one or more other carbocyclic aromatic rings or heteroaromatic rings. Thus, "5 - to 14 - membered heteroaryl" includes monocyclic systems, bicyclic systems, or tricyclic ring systems.

[0039] Examples of monocyclic 5- or 6-membered heteroaryl groups include furanyl (e.g., 2-furanyl, 3-furanyl), imidazolyl (e.g., N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl), isoxazolyl (e.g., 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl), oxadiazolyl (e.g., 2-oxadiazolyl, 5-oxadiazolyl), oxazolyl (e.g., 2-oxazolyl, 4-oxazolyl, 5-oxazolyl), pyrazolyl (e.g., 3-pyrazolyl, 4-pyrazolyl), pyrrolyl (e.g., 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl), pyridyl (e.g., 2-pyridyl, 3-pyridyl, 4-pyridyl), pyrimidinyl (e.g., 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl), pyridazinyl (e.g., 3-pyridazinyl), thiazolyl (e.g., 2-thiazolyl, 4-thiazolyl, 5-thiazolyl), triazolyl (e.g., 2-triazolyl, 5-triazolyl), tetrazolyl (e.g., tetrazolyl), thienyl (e.g., 2-thienyl, 3-thienyl), pyrimidinyl, pyridinyl and pyridazinyl. Examples of polycyclic aromatic heteroaryl groups include carbazolyl, benzimidazolyl, benzothienyl, benzofuranyl, indolyl, quinolinyl, benzotriazolyl, benzothiazolyl, benzoxazolyl, benzimidazolyl, isoquinolinyl, indolyl, isoindolyl, acridinyl, or benzisoxazolyl. A "substituted heteroaryl group" is substituted with any one or more substitutable ring atoms that are ring carbon or ring nitrogen atoms bonded to hydrogen.

[0040] The term "bridged bicyclic group" refers to a ring system that includes two rings sharing at least three adjacent ring atoms.

[0041] As used herein, many moieties (e.g., alkyl, alkylene, cycloalkyl, aryl, heteroaryl, or heterocyclyl) are referred to as either "substituted" or "optionally substituted". When a moiety is modified by one of these terms, unless otherwise noted, it means that any portion of the moiety known to those skilled in the art as being available for substitution may be substituted, and it includes one or more substituents. If more than one substituent is present, each substituent may be independently selected. Such means for substitution are well known in the art and / or are taught by the present disclosure. An optional substituent may be any substituent suitable for attachment to the moiety.

[0042] When suitable substituents are not specifically listed, exemplary substituents include, for example, (C1-C5)alkyl, (C1-C5)hydroxyalkyl, (C1-C5)haloalkyl, (C1-C5)alkoxy, (C1-C5)haloalkoxy, halogen, hydroxyl, cyano, amino, -CN, -NO2, -OR c1 、-NR a1 R b1 、-S(O) i R a1 、-NR a1 S(O) i R b1 、-S(O) i NR a1 R b1 、-C(=O)OR a1 、-OC(=O)OR a1 、-C(=S)OR a1 、-O(C=S)R a1 、-C(=O)NR a1 R b1 、-NR a1 C(=O)R b1 、-C(=S)NR a1 R b1 、-C(=O)R a1 、-C(=S)R a1 、NR a1 C(=S)R b1 、-O(C=O)NR a1 R b1 、-NR a1(C=S)OR b1 、 -O(C=S)NR a1 R b1 、 -NR a1 (C=O)NR a1 R b1 、 -NR a1 (C=S)NR a1 R b1 、 phenyl, or 5- to 6-membered heteroaryl, including but not limited to these. Each R a1 and each R b1 is independently selected from -H and (C1-C5)alkyl, and optionally substituted with hydroxyl or (C1-C3)alkoxy. R c1 is -H, (C1-C5)haloalkyl or (C1-C5)alkyl, and (C1-C5)alkyl is optionally substituted with hydroxyl or (C1-C3)alkoxy.

[0043] The compounds described herein can contain one or more chiral centers and, thus, can exist in various stereoisomers, such as enantiomers and / or diastereomers. For example, the compounds described herein can be in the form of individual enantiomers, diastereomers or geometric isomers, or in the form of mixtures of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers.

[0044] Enantiomeric and diastereomeric mixtures can be resolved into their component enantiomers or stereoisomers by well-known methods such as chiral phase gas chromatography, chiral phase high performance liquid chromatography, crystallizing the compound as a chiral salt complex, or crystallizing the compound in a chiral solvent. Enantiomers and diastereomers can also be obtained from diastereomerically or enantiomerically pure intermediates, reagents, and catalysts by well-known asymmetric synthesis methods. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, E.L. Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, S.H. Tables of Resolving Agents and Optical Resolutions p.268 (E.L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972).

[0045] When a compound is designated by a name or structure that indicates a single enantiomer, the compound is at least 60%, 70%, 80%, 90%, 99% or 99.9% optically pure (also called "enantiomerically pure") unless otherwise specified. Optical purity is the weight of the named or depicted enantiomer of the mixture divided by the total weight of the mixture of both enantiomers.

[0046] The stereochemistry of the disclosed compounds is named or depicted by the structure, and if the named or depicted structure encompasses more than one stereoisomer (such as in the case of a pair of diastereomers), it is to be understood that one of the included stereoisomers or any mixture of the included stereoisomers is included. It is further to be understood that the stereoisomeric purity of the named or depicted stereoisomer is at least 60 wt%, 70 wt%, 80 wt%, 90 wt%, 99 wt%, or 99.9 wt%. The stereoisomeric purity in this case is determined by dividing the total weight of the mixture of stereoisomers included in the name or structure by the total weight of the mixture of all stereoisomers. is determined by dividing the total weight of the mixture of stereoisomers included in the name or structure by the total weight of the mixture of all stereoisomers.

[0047] When geometric isomers are depicted by the name or structure, it is to be understood that the geometric isomeric purity of the named or depicted isomer is at least 60 wt%, 70 wt%, 80 wt%, 90 wt%, 99 wt%, or 99.9 wt% pure. The geometric isomeric purity is determined by dividing the weight of the named or depicted geometric isomer in the mixture by the total weight of all geometric isomers in the mixture.

[0048] A racemic mixture means 50% of one enantiomer and 50% is the corresponding enantiomer. The present invention encompasses all enantiomerically pure, enantiomerically enriched, diastereomerically pure, diastereomerically enriched, and racemic mixtures of the compounds of the present invention, as well as diastereomeric mixtures.

[0049] The compounds described herein may also include all isotopes of atoms present in the intermediates or final compounds. Isotopes include atoms that have the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium.

[0050] It will be recognized that depending on the origin of the chemical substances used in the synthesis, some variation in the natural isotope abundances will occur in the synthesized compounds. Thus, preparations of the compounds disclosed herein will essentially contain minor amounts of deuterated isotopologs. Despite this variation, the concentrations of the naturally abundant and stable hydrogen and carbon isotopes are low and not significant compared to the degree of stable isotope substitution of the compounds of the invention. See, for example, Wada, E et al., Seikagaku, 1994, 66:15; Gannes, LZ et al., Comp Biochem Physiol Mol Integr Physiol, 1998, 119:725.

[0051] The compounds described herein may exist in various tautomeric forms. The terms "tautomer" or "tautomeric" refer to two or more interconvertible compounds / substituents resulting from at least one formal shift of a hydrogen atom and at least one change in valence (e.g., from a single bond to a double bond, from a triple bond to a single bond, or vice versa). Exemplary tautomerizations include the tautomerization from keto to enol, from amide to imide, from lactam to lactim, from enamine to imine, and from enamine to (a different enamine). The teachings of the present invention encompass compounds in tautomeric forms, including forms not structurally depicted. All such isomeric forms of such compounds are explicitly included. If a tautomer of a compound is aromatic, the compound is aromatic. Similarly, if a tautomer of a compound is heteroaryl, the compound is heteroaryl.

[0052] In certain examples, there exist tautomeric forms of the disclosed compounds, such as the tautomeric structures shown below.

Chemical Structure

[0053] When the compounds of this specification are represented by a structural formula or designated by a chemical name in this specification, all other possible tautomeric forms that may exist for that compound are included in the structural formula. It is to be understood that it can be.

[0054] The compounds of the present invention can exist in free form for therapeutic use or, where appropriate, in pharmaceutically acceptable salt form.

[0055] The term "pharmaceutically acceptable salt" refers to salts that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, etc., and that have a reasonable benefit / risk ratio commensurate with such use. Pharmaceutically acceptable salts are well known in the art; for example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, which is incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable acid addition salts are salts of amino groups formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by using other methods known in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Salts derived from appropriate bases include alkali metal salts, alkaline earth metal salts, ammonium salts, and N + (C 1-4 alkyl)4 -Salts are included. Representative alkali metal salts or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Further pharmaceutically acceptable salts include, where appropriate, ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates.

[0056] Such pharmaceutically acceptable acid addition salts and general methodologies for preparing them are well known in the art. For example, see Stahl et al., HANDBOOK OF PHARMACEUTICAL SALTS: PROPERTIES, SELECTION AND USE, (VCHA / Wiley-VCH, 2002); Bighley et al., in “Encyclopedia of Pharmaceutical Technology.” Eds. Swarbrick and Boylan, Vol. 13, Marcel Dekker, Inc., New York, Basel, Hong Kong 1995, pp. 453~499; Berge et al., “Pharmaceutical Salts,” Journal of Pharmaceutical Sciences, 66(1):1977.

[0057] The terms “composition” and “formulation” are used interchangeably.

[0058] The “subject” is a mammal, preferably a human, but may also be an animal in need of veterinary treatment, such as a companion animal (e.g., dog, cat, etc.), a farm animal (e.g., cow, sheep, pig, horse, etc.) and a laboratory animal (e.g., rat, mouse, guinea pig, etc.).

[0059] The terms "administer", "administering", or "administration" refer to methods of introducing the compounds of the invention or compositions thereof into or onto a subject. These methods include, but are not limited to, intra-articular (within the joint), intravenous, intramuscular, intratumoral, intradermal, intraperitoneal, subcutaneous, oral, topical, intrathecal, inhalation, transdermal, rectal, etc. Administration techniques that can be used with the agents and methods described herein can be found, for example, in Goodman and Gilman, The Pharmacological Basis of Therapeutics, current ed.; Pergamon; and Remington’s, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pa.

[0060] The terms "treat", "treating", and "treatment" refer to reversing, alleviating, or inhibiting the progression of a disease described herein. In some embodiments, treatment can be administered after one or more signs or symptoms of the disease have occurred or been observed (i.e., treatment for treatment). In other embodiments, treatment can be administered in the absence of signs or symptoms of the disease. For example, treatment can be administered to a susceptible subject prior to the onset of symptoms (i.e., treatment for prevention) (e.g., taking into account the symptom history and / or exposure to a pathogen). Treatment can also continue after the symptoms have resolved, for example, to delay or prevent recurrence.

[0061] The terms "condition", "disease", and "disorder" are used interchangeably.

[0062] Generally, the effective amount of the compounds taught herein will vary depending on various factors such as the particular drug or compound, pharmaceutical formulation, route of administration, type of disease or disorder, identity of the subject or host being treated, etc., but nonetheless can be routinely determined by one of ordinary skill in the art. The effective amount of the compounds of the present teachings can be readily determined by one of ordinary skill in the art using routine methods known in the art.

[0063] The term "effective amount" means an amount that, when administered to a subject, provides a beneficial or desirable result, including, for example, inhibiting, suppressing or reducing clinical outcomes, such as symptoms of a condition being treated in the subject, as compared to a control. For example, an effective amount can be administered in a unit dosage form (e.g., from 1 mg to about 50 g per day, such as from 1 mg to about 5 grams per day).

[0064] A "therapeutically effective amount" is an amount effective in the detectable killing or inhibition of cancer cell growth or spread; tumor size or number; or other measures of cancer level, stage, progression or severity. The exact amount required will vary for each subject depending on species, age, and general condition of the subject, severity of the disease, particular anti-cancer agent, its mode of administration, combination therapy with other treatments, etc.

[0065] The common chemical terms used in the above formula have their ordinary meanings.

[0066] As used herein, "h" refers to 1 hour or several hours, "min" refers to several minutes or minutes, "Cdk" or "CDK" refers to cyclin-dependent kinase, "pRb" refers to retinoblastoma protein, "MCL" refers to mantle cell lymphoma, "AML" refers to acute myeloid leukemia, "CML" refers to chronic myeloid leukemia, "Boc" refers to N-tert-butoxycarbonyl, "EA" refers to ethyl acetate, "DCM" refers to dichloromethane, "DMSO" refers to dimethyl sulfoxide, "DMA" refers to di Refers to methylacetamide, "THF" refers to tetrahydrofuran, "MtBE" refers to methyl tert-butyl ether, "TEA" refers to triethylamine, "FBS" refers to fetal bovine serum, "PBS" refers to phosphate buffered saline, "BSA" refers to bovine serum albumin, "RT" refers to room temperature, "mpk" means milligrams per kilogram, "po" refers to per os (oral), "qd" means once-a-day administration, "HPLC" means high performance liquid chromatography, "q2d" means single administration every two days, "q2dx10" means single administration every two days × 10 times, "VSMC" means vascular smooth muscle cells, and "XRD" refers to X-ray diffraction.

[0067] 3. Compound In the first embodiment of the present invention, a compound represented by the structural formula (I)

Chemical formula

Chemical formula

[0068] In a second embodiment of the present invention, there is provided a compound represented by structural formula (I), or a pharmaceutically acceptable salt or stereoisomer thereof, wherein ring C is

Chemical formula

[0069] In a third embodiment of the present invention, there is provided a compound of the first or second embodiment, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein ring A is

Chemical formula

[0070] In a fourth embodiment of the present invention, the compound of structural formula (I) is represented by structural formulas (II-A) to ( II-J)

Chemical formula

[0071] In a fifth embodiment of the present invention, there are provided compounds of structural formula (I), (II-A) to (II-J), or a pharmaceutically acceptable salt or stereoisomer thereof, wherein L is a bond, -(CH2)-, -O(CH2)-, -C(=O)-, or -S(O)2-, and the remaining variables are as defined in the first, second, third, and / or fourth embodiments.

[0072] In a sixth embodiment of the present invention, structural formula (I), (II-A) to (II-J) compounds, or pharmaceutically acceptable salts or stereoisomers thereof are provided, wherein ring B is a 4- to 10-membered heterocyclyl or a 5- to 6-membered monocyclic heteroaryl optionally substituted with one or two R2 groups, and the remaining variables are as defined in the First, Second, Third, Fourth, and / or Fifth Embodiments.

[0073] In the seventh embodiment of the present invention, structural formula (I), (II-A) to (II-J) compounds, or pharmaceutically acceptable salts or stereoisomers thereof are provided, wherein each occurrence of R2 is H, halogen, CN, -OH, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 hydroxyalkyl, -(CH2) n OR6, -(CH2) n C(O)R6, -(CH2) n C(O)OR6, -(CH2) n S(O)2R6, -(CH2) n NR7R8, -(CH2) n C(O)NR7R8, -(CH2) n C(O)NHR7, -(CH2) n NR7C(O)R6, -(CH2) n NR7S(O)2R6, C 3-8 cycloalkyl, a 3- to 6-membered heterocyclyl, phenyl, or a 5- to 6-membered heteroaryl; or two R2s attached to the same ring atom of ring B form a 3- to 6-membered heterocyclyl (when ring B is a 3- to 10-membered heterocyclyl) optionally substituted with one or more groups selected from halogen, -OH, C alkyl, C 1-2 alkyl, C 1-2 haloalkyl, C 1-2 alkoxy, C 1-2 haloalkoxy, and NR7R8; each occurrence of R6 is independently H, C 1-4 alkyl, C 3-6Cycloalkyl, 3- to 7-membered heterocyclyl, phenyl, 5- to 6-membered heteroaryl, and C represented by R6 1-4 alkyl, C 3-6 cycloalkyl, 3- to 7-membered heterocyclyl, phenyl, 5- to 6-membered heteroaryl are each optionally substituted with halogen, CN, -OH, C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, or NR7R8; n is 0, 1, or 2, and the remaining variables are as defined in the first, second, third, fourth, fifth, and / or sixth embodiments.

[0074] In the eighth embodiment of the present invention, compounds of structural formula (I), (II-A) to (II-J), or a pharmaceutically acceptable salt or stereoisomer thereof, are provided, wherein R1 is H, F, Cl, or CH3, and the remaining variables are as defined in the first, second, third, fourth, fifth, sixth, and / or seventh embodiments.

[0075] In the ninth embodiment of the present invention, compounds of structural formula (I), (II-A) to (II-J), or a pharmaceutically acceptable salt or stereoisomer thereof, are provided, wherein ring B is

Chemical formula

[0076] In the tenth embodiment of the present invention, compounds of structural formula (I), (II-A) to (II-J), or a pharmaceutically acceptable salt or stereoisomer thereof, are provided, wherein each occurrence of R2 is H, halogen, CN, -OH, C 1-4 alkyl, C 1-4 haloalkyl, C1-4 Hydroxyalkyl, -(CH2) n S(O)2C 1-4 alkyl, -(CH2) n NR7R8, C 3-4 cycloalkyl, or a 3- to 6-membered heterocyclyl; n is 0, 1, or 2; or when ring B is a 4- to 7-membered heterocyclyl, two R2s attached to the same ring atom of ring B form a 3- to 6-membered heterocyclyl, and the remaining variables are as defined in the first, second, third, fourth, fifth, sixth, seventh, eighth, and / or ninth embodiments.

[0077] In the eleventh embodiment of the present invention, the structural formula (I), (II-A) to (II-J) compounds, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein ring A is

Chemical formula

[0078] In the twelfth embodiment of the present invention, the structural formula (I), (II-A) to (II-J) compounds, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein R1 is H or F, and the remaining variables are as defined in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, and / or eleventh embodiments.

[0079] In the thirteenth embodiment of the present invention, the structural formula (I), (II-A) to (II-J) compounds, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein each occurrence of R3 is H, C optionally substituted with -OH 1-3 alkyl, or C optionally substituted with -OH 3-6is cycloalkyl; each occurrence of R4 is H, halogen, C 1-3 alkyl, C 2-4 alkenyl, cyclopentyl, tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl; each occurrence of R5 is H, F, CN, methoxy, OCHF2, and the remaining variables are as defined in the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, and / or 12th embodiments.

[0080] In a 14th embodiment of the present invention, there is provided a compound of structural formula (I), (II-A) to (II-J), or a pharmaceutically acceptable salt or stereoisomer thereof, wherein L is a bond and the remaining variables are as defined in the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, and / or 13th embodiments.

[0081] In a 15th embodiment of the present invention, there is provided a compound of structural formula (I), (II-A) to (II-J), or a pharmaceutically acceptable salt or stereoisomer thereof, wherein ring B is

Chemical formula

[0082] In a 16th embodiment of the present invention, there is provided a compound of structural formula (I), (II-A) to (II-J), or a pharmaceutically acceptable salt or stereoisomer thereof, wherein each occurrence of R2 is H, halogen, CN, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 hydroxyalkyl, NH2, N(CH3)2, NH cyclopropyl, -(CH2) n S(O)2C1-3 alkyl, cyclopropyl, azetidinyl, oxetanyl, morpholinyl, piperidinyl, tetrahydro-2H-pyranyl optionally substituted with F, or when ring B is piperidinyl, two R2s attached to the same ring atom of ring B form 2,5-pyrrolidinedionyl, or 2-pyrrolidonyl, n is 0, 1, or 2, and the remaining variables are as defined in the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, and / or 15th embodiments.

[0083] In the 17th embodiment of the present invention, the structural formula (I), (II-A) to (II-J) compounds, or pharmaceutically acceptable salts or stereoisomers thereof are provided, wherein ring A is

Chemical formula

[0084] In the 18th embodiment of the present invention, the structural formula (I), (II-A) to (II-J) compounds, or pharmaceutically acceptable salts or stereoisomers thereof are provided, wherein each occurrence of R3 is H or C 1-3 alkyl, and each occurrence of R4 is H or C 1-3 alkyl, and each occurrence of R5 is H, F, or OMe, and the remaining variables are as defined in the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, and / or 17th embodiments.

[0085] In the 19th embodiment of the present invention, the structural formula (I) is Compounds of (II-A) to (II-J), or pharmaceutically acceptable salts or stereoisomers thereof are provided, wherein R1 is H and the remaining variables are as defined in the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, and / or 18th embodiments.

[0086] In the 20th embodiment of the present invention, the structural formula (I), (II-A) to (II-J), or pharmaceutically acceptable salts or stereoisomers thereof are provided, wherein ring C is unsubstituted and the remaining variables are as defined in the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, and / or 19th embodiments.

[0087] In the 21st embodiment of the present invention, the structural formula (I), Compounds of (II-A) to (II-J), or pharmaceutically acceptable salts or stereoisomers thereof are provided, wherein ring B is

Chemical formula

[0088] In the 22nd embodiment of the present invention, compounds of the structural formula (I), (II-A) to (II-J), or pharmaceutically acceptable salts or stereoisomers thereof are provided, wherein R2 is H, halogen, CN, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3It is hydroxyalkyl, NH2, N(CH3)2, NH cyclopropyl, and the remaining variables are as defined in the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 16th, 17th, 18th, 19th, 20th, and / or 21st embodiments.

[0089] In one embodiment, the compound or its pharmaceutically acceptable salt, or stereoisomer, is selected from the compounds of formula (I), (II-A) to (II-J) of the examples.

[0090] Another aspect of the present disclosure relates to the labeled compounds (radioactive labels, fluorescent labels, etc.) of the present invention, which are useful not only in imaging techniques but also in assays for localizing and quantifying CDK in tissue samples including humans, both in vitro and in vivo, and for identifying CDK ligands by inhibiting the binding of labeled compounds. Accordingly, the present disclosure includes such labeled compounds.

[0091] The present disclosure further includes the isotope-labeled compounds of the present invention. "Isotopic" or "radioactively labeled" compounds are compounds of the present invention in which one or more atoms are replaced by or substituted with atoms having an atomic weight or mass number different from that typically found in nature (i.e., of natural origin). Suitable radionuclides that can be incorporated into the compounds of the present invention are 2 H (also denoted as D in the case of deuterium), 3 H (also denoted as T in the case of tritium), 11 C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 O, 18 F, 35 S, 36 Cl, 82 Br, 75 Br, 76 Br, 77 Br, 123 I, 124 I, 125I and 131 including, but not limited to, I. The radionuclide incorporated into the radiolabeled compound of the present invention depends on the specific use of the radiolabeled compound.

[0092] The present invention can further include a synthesis method for incorporating a radioisotope into the compound of the present invention. Synthesis methods for incorporating radioisotopes into organic compounds are well known in the art, and those skilled in the art will readily recognize methods applicable to the compounds of the present invention.

[0093] The labeled compound of the present invention can be used in screening assays for identifying / evaluating compounds. For example, a newly synthesized or identified labeled compound (i.e., a test compound) can be evaluated for its ability to bind to CDK by monitoring its concentration fluctuations when in contact with CDK through tracking the label. For example, a test compound (labeled) can be evaluated for its ability to reduce the binding of another compound (i.e., a standard compound) known to bind to CDK. Thus, the ability of a test compound to compete with a standard compound for binding to CDK is directly correlated with its binding affinity. Conversely, in some other screening assays, the standard compound is labeled and the test compound is not. Thus, to evaluate the competition between the standard compound and the test compound, the concentration of the labeled standard compound is monitored to confirm the relative binding affinity of the test compound. In one embodiment, a compound or a pharmaceutically acceptable salt or stereoisomer thereof in which one or more hydrogen atoms are replaced by deuterium is provided.

[0094] In one embodiment, a compound or a pharmaceutically acceptable salt or stereoisomer thereof in which one or more hydrogen atoms are replaced by deuterium is provided.

[0095] 4. Treatable Diseases and Treatments The specific compounds of the present invention are selective inhibitors of CDK2, CDK4, and / or CDK6, and are thus useful for the treatment of diseases or disorders characterized by abnormal cell proliferation that can be inhibited by a decrease in the activity of CDK-cyclin complexes including CDK2, CDK4, and / or CDK6.

[0096] In certain embodiments, the compounds of the present invention selectively inhibit CDK4 / 6 over CDK2, and the ratio of the IC 50 value for the former (CDK4 / 6) to the latter (CDK2) is at least about 10, 20, 50, 100, 200, 300, 400, 500, 800, 1,000, 2,000 or more.

[0097] In certain embodiments, the compounds of the present invention selectively inhibit CDK4 over CDK6, and the ratio of the IC 50 value for the former (CDK4) to the latter (CDK6) is at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 50 or more.

[0098] In certain embodiments, the compounds of the present invention selectively inhibit CDK2 over CDK4, and the ratio of the IC 50 value for the former (CDK2) to the latter (CDK4) is at least about 2, 5, 10, 15, 20, 40, 50, 60, 80, 100 or more.

[0099] In certain embodiments, the compounds of the present invention inhibit CDK2 / 4 / 6 with similar IC 50 values, e.g., within 10-fold, 5-fold, 3-fold, or 2-fold of the IC 50 value. Such compounds of the present invention are useful for treating cancers associated with amplified or enhanced expression of cyclin D1 or E1 or E2.

[0100] CDK2 is the catalytic subunit of a CDK-cyclin complex whose activity is restricted to the G1-S phase of the cell cycle, where the cell makes the proteins necessary for mitosis and replicates its DNA. CDK2 forms a complex with cyclin E or A. Cyclin E binds to CDK2 in the G1 phase, which is required for the transition from the G1 to the S phase. On the other hand, CDK2 that binds to cyclin A is required to proceed through the S phase.

[0101] CDK2 is largely dispensable in the cell cycle of normal functioning cells but is important in the abnormal proliferation process of cancer cells. Overexpression of cyclin E occurs in many tumor cells, causing the cells to be dependent on CDK2 and cyclin E. Abnormal cyclin E activity is observed in breast, lung, colorectal, gastric, and bone cancers, as well as leukemia and lymphoma. Similarly, abnormal expression of cyclin A2 is associated with chromosomal instability and tumor growth, and inhibition leads to a decrease in tumor growth. Therefore, CDK2 and its cyclin-binding partners are potential therapeutic targets for new cancer therapeutics. Preclinical models have shown preliminary success in restricting tumor growth and have also been observed to reduce the side effects of current chemotherapeutic drugs.

[0102] For example, Caldon et al. (Mol Cancer Ther 11(7):1488-1499, 2012) reported that cyclin E2 is included in several gene signatures that predict disease progression in either tamoxifen-resistant or metastatic breast cancer, that high expression of CycE2 is characteristic of the luminal B and HER2 subtypes of breast cancer, and that it strongly predicts a shortened survival without distant metastasis after endocrine therapy. Furthermore, tamoxifen-resistant (MCF-7 TAMR) breast cancer cells overexpress cyclin E2, and expression of either cyclin E1 or E2 in T-47D breast cancer cells conferred acute anti-estrogen resistance, suggesting that overexpression of cyclin E contributes to anti-estrogen resistance in tamoxifen-resistant cells. Proliferation of tamoxifen-resistant cells was inhibited by knockdown via RNAi of cyclin E1, cyclin E2, or CDK2. In addition, ectopic expression of cyclin E1 or E2 also decreased sensitivity to inhibition of CDK4 but not CDK2. Furthermore, inhibition of CDK2 in E-cyclin overexpressing cells and tamoxifen-resistant cells restored sensitivity to tamoxifen or CDK4 inhibition.

[0103] These data indicate that overexpression of cyclin E2 is a potential mechanism of resistance to both endocrine therapy and CDK4 inhibition, and that CDK2 inhibitors can effectively inhibit cells overexpressing cyclin E1 and E2, overcome such resistance, and be beneficial as a component of combination therapy for endocrine-resistant diseases, as they enhance the effectiveness of other therapeutic agents. Similarly, the compounds of the subject having potent inhibitory activity against both CDK2 and CDK4 are expected to be effective against cancer cells that are either non-resistant or resistant to endocrine therapy or CDK4 inhibition.

[0104] Thus, in certain embodiments, the compounds of the invention can have a potent inhibitory effect against both CDK2 and CDK4 (e.g., independently, <10 nM, <5 nM, <1 nM levels of IC 50value), and thus is effective for the treatment of tamoxifen-resistant or metastatic breast cancer such as tamoxifen-resistant or metastatic breast cancer associated with overexpression of CycE.

[0105] The IC of the compound of the present invention against CDK2 / 4 / 6 50 value can be measured, for example, using the methods described in Examples 1 to 3 (incorporated herein by reference).

[0106] In particular, the compounds of the present invention are useful for the treatment of cancer. In other embodiments, the compounds of the present invention are useful for the treatment of chronic inflammatory diseases such as arthritis and cystic fibrosis.

[0107] Thus, in one aspect, the present invention provides a method for treating cancer, particularly the cancers described herein, in a mammal, the method comprising administering to a mammal in need of such treatment an effective amount of a compound of the present invention.

[0108] In a related aspect, the present invention relates to the use of a compound of the present invention in the manufacture of a medicament for treating cancer, particularly the cancers described herein.

[0109] In another related aspect, the compounds of the present invention can be used in the manufacture of a medicament for the treatment of cancer, particularly the cancers described herein.

[0110] In another related aspect, the present invention provides a compound of the present invention for use in the treatment of cancer, particularly the cancers described herein.

[0111] According to any of the above related aspects of the present invention, CDK4 and CDK6 can regulate their effects on the cell cycle, at least in part, through phosphorylation of pRb. Thus, the specific compounds of the present invention can inhibit pRb phosphorylation by inhibiting CDK4 / 6 activity in any cancer type in which the cells are proliferating and contain a functional and intact Rb1 gene encoding pRb, and thus can inhibit cell proliferation and / or tumor growth.

[0112] Thus, in certain embodiments, the compounds of the invention are pRb in mammals + cancer, such as colorectal cancer, breast cancer, lung cancer, prostate cancer, chronic myelogenous leukemia, acute myelogenous leukemia (Fry et al., Mol. Cancer Ther. 3(11):1427, 2004), mantle cell lymphoma (Marzec et al., Blood 108(5):1744, 2006), ovarian cancer ((Kim et al., Cancer Research 54:605, 1994), pancreatic cancer (Schutte et al., Cancer Research 57:3126, 1997), malignant melanoma and metastatic malignant melanoma (Maelandsmo et al., British Journal of Cancer 73:909, 1996). The compounds of the invention are also useful in the treatment of rhabdomyosarcoma (Saab et al., Mol. Cancer. Ther. 5(5):1299, 2006) in mammals (e.g., humans), and multiple myeloma including relapsed and refractory multiple myeloma (Baughn et al., Cancer Res. 66(15):7661, 2006).

[0113] On the other hand, Zhang et al. (Nature dx.doi.org / 10.1038 / nature25015, 2017) showed that inhibition of CDK4 / 6 in vivo decreased phosphorylation, and thus (by APC / C Cdh1 Cullin 3 SPOPThe degradation of E3 ligase may increase, which has been reported to result in an increase in the PD-L1 level on the surface of tumor cells and a decrease in the number of tumor-infiltrating lymphocytes (TIL) in mouse tumors and primary human prostate cancer specimens. In other words, in vivo inhibition of CDK4 / 6 increases the PD-L1 protein level and contributes to an increase in resistance to immune checkpoint therapy targeting PD-1 (programmed cell death protein 1) and PD-L1 (the ligand of PD-1). On the other hand, combining CDK4 / 6 inhibitor treatment with anti-PD-1 immunotherapy promotes tumor regression and dramatically improves the overall survival rate in mouse tumor models.

[0114] Accordingly, in certain embodiments, the compounds of the present invention can be used in combination with PD-1 / PD-L1 immune checkpoint inhibitors to enhance the therapeutic effect on human cancer.

[0115] The PD-1 and PD-L1 inhibitors that can be used with the compounds of the present invention are known in the art. PD-1 inhibitors include monoclonal antibodies specific to PD-1 or antigen-binding fragments thereof. Exemplary PD-1 inhibitors include pembrolizumab (Keytruda), nivolumab (Opdivo), and cemiplimab (Libtayo). PD-L1 inhibitors include monoclonal antibodies specific to PD-L1 or antigen-binding fragments thereof. Exemplary PD-L1 inhibitors include atezolizumab (Tecentriq), avelumab (Bavencio), and durvalumab (Imfinzi).

[0116] Additional immune checkpoint inhibitors that can be used with the compounds of the present invention to enhance the therapeutic effect on human cancer include monoclonal antibodies specific to CTLA-4, such as ipilimumab (Yervoy), or antigen-binding fragments thereof.

[0117] Additional immune checkpoint inhibitors that can be used in combination with the compounds of the present invention to enhance the therapeutic effect against human cancer include bispecific monoclonal antibodies specific for PD-1 and PD-L1 or antigen-binding fragments thereof, or monoclonal antibodies specific for PD-1 and PD-L1, or combinations of monoclonal antibodies or antigen-binding fragments thereof specific for PD-1 and CTLA-4, etc.

[0118] In certain embodiments, the compounds of the present invention can be used in combination with a Tyr kinase inhibitor, for example, a receptor Tyr kinase (RTK) inhibitor, to enhance the therapeutic effect against human cancer. Exemplary Tyr kinase inhibitors include ALK inhibitors (crizotinib, ceritinib, alectinib, brigatinib, etc.), Bcr-Abl inhibitors (bosutinib, dasatinib, imatinib, nilotinib, ponatinib, etc.), BTK inhibitors (ibrutinib, etc.), c-Met inhibitors (crizotinib, cabozantinib, etc.), EGFR inhibitors (gefitinib, erlotinib, lapatinib, vandetanib, afatinib, osimertinib, etc.), JAK inhibitors (ruxolitinib, tofacitinib, etc.), MEK1 / 2 inhibitors (trametinib, etc.), PDGFR inhibitors (axitinib, gefitinib, imatinib, lenvatinib, nintedanib, pazopanib, regorafenib, sorafenib, sunitinib, etc.), RET inhibitors (vandetanib, etc.), Src family kinase inhibitors (bosutinib, dasatinib, ponatinib, vandetanib, etc.), and VEGFR family inhibitors (axitinib, lenvatinib, nintedanib, regorafenib, pazopanib, sorafenib, sunitinib, etc.).

[0119] Additional suitable kinase inhibitors that can be used in combination with the compounds of the subject matter, as well as the cancer indications treatable, are described in Bhullar et al., Molecular Cancer 17:48, 2018 (incorporated herein by reference in its entirety).

[0120] Additional additional RTK inhibitors include monoclonal antibodies and antigen-binding fragments thereof, including anti-EGFR mAbs such as cetuximab (e.g., effective in the treatment of lung cancer, colorectal cancer, and head and neck cancer), and anti-HER2 mAbs such as trastuzumab (e.g., effective in the treatment of breast cancer).

[0121] In certain embodiments, the compounds of the invention can be used in combination with antagonists of hormone receptor signaling such as those previously described for the treatment of breast cancer.

[0122] Cancers treatable with the compounds of the invention include: non-Hodgkin lymphoma; malignant mesothelioma; non-small cell lung cancer; cholangiocarcinoma; soft tissue sarcoma; glioblastoma; (recurrent) brain tumor; brain metastases secondary to hormone receptor-positive breast cancer, non-small cell lung cancer, melanoma (including cyclin D1-expressing positive melanoma); (recurrent or persistent) endometrial cancer; (recurrent or metastatic) head and neck squamous cell carcinoma (HNSCC); hepatocellular carcinoma; esophageal squamous cell carcinoma (SCC); esophageal adenocarcinoma (ADC); renal cell carcinoma, and urothelial carcinoma.

[0123] In certain embodiments, treatable cancers include: cancer of the bladder, breast, colon, kidney, epidermis, liver, lung (including SCLC and NSCLC), esophagus, gallbladder, ovary, pancreas, stomach, cervix, thyroid, nose, head and neck, prostate, or skin; lymphoid hematopoietic tumors; myeloid hematopoietic tumors; thyroid follicular cancer; tumors of mesenchymal origin; tumors of the central or peripheral nervous system; melanoma; familial melanoma; seminoma; teratocarcinoma; osteosarcoma; xeroderma pigmentosum; corneal tumor; thyroid follicular cancer; Kaposi sarcoma, squamous cell carcinoma, sarcoma; or tumors of mesenchymal origin.

[0124] In certain embodiments, lymphoid hematopoietic tumors are leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, B-cell lymphoma, T-cell lymphoma, multiple myeloma, Hodgkin lymphoma, non-Hodgkin lymphoma, hairy cell lymphoma, or Burkitt lymphoma.

[0125] In certain embodiments, the tumor of the central nervous system or peripheral nervous system is astrocytoma, neuroblastoma, glioma or schwannoma.

[0126] In certain embodiments, the cancer is small cell lung cancer, non-small cell lung cancer, pancreatic cancer, breast cancer, glioblastoma multiforme, T cell ALL and mantle cell lymphoma.

[0127] In certain embodiments, the cancer is selected from the group consisting of colorectal cancer, mantle cell lymphoma, breast cancer (including advanced or metastatic or recurrent breast cancer), pancreatic cancer, ovarian cancer, glioblastoma, acute myeloid leukemia, and lung cancer, particularly NSCLC.

[0128] In certain embodiments, the cancer is NSCLC, pancreatic cancer, ovarian cancer or metastatic breast cancer, and the treatment comprises administering to a mammal in need thereof a therapeutically effective combination of a compound of the invention and gemcitabine HCl.

[0129] In certain embodiments, the cancer is NSCLC, pancreatic cancer, ovarian cancer or metastatic breast cancer, and the drug comprising the compound of the invention also comprises gemcitabine HCl or is administered simultaneously with, separately from, or sequentially to gemcitabine HCl.

[0130] In certain embodiments, the compounds of the invention can be used in combination with other agents for the treatment of NSCLC, pancreatic cancer, ovarian cancer and metastatic breast cancer. For example, the compounds of the invention can be used in simultaneous, separate or sequential combination with gemcitabine HCl in the treatment of NSCLC, pancreatic cancer, ovarian cancer or metastatic breast cancer.

[0131] In certain embodiments, the cancer is selected from the group consisting of colorectal cancer, glioblastoma, acute myeloid leukemia and lung cancer.

[0132] In certain embodiments, the cancer is glioblastoma or astrocytoma, and the treatment utilizes a therapeutically effective combination of a compound of the invention and temozolomide. The compound of the invention can be administered simultaneously with, separately from, or sequentially to temozolomide.

[0133] Treatment of breast cancer In certain embodiments, the compounds of the invention can be used to treat breast cancer.

[0134] Breast cancer is a significant health burden worldwide, accounting for approximately 7% of all cancer-related deaths in the United States in 2016 alone. Of all breast cancers, approximately 75% are diagnosed as hormone receptor-positive (HR + ) breast cancer, which expresses estrogen receptor (ER) and / or progesterone receptor (PgR) and typically relies on the ER signaling pathway for growth and survival. That is, HR + breast cancer utilizes the biological functions of the ER pathway to promote the growth, development, and progression of breast cancer. On the other hand, since HR + breast cancer depends on ER signaling, such breast cancer is a therapeutic target for endocrine therapy agents that target the estrogen signaling pathway, such as aromatase inhibitors (AIs; including letrozole, anastrozole, and exemestane), selective ER modulators (tamoxifen), and selective ER downregulators (fulvestrant).

[0135] Endocrine therapy constitutes the treatment backbone for HR + breast cancer, but the effectiveness of endocrine therapy is limited because the proportion of both existing and newly acquired resistance during treatment due to the presence of alternative survival or "escape" pathways is high. Many of the ER pathway and known escape pathways act through cyclin D-CDK4 / 6-inhibitors of the CDK4(INK4)-retinoblastoma (Rb) pathway to promote tumor growth. Therefore, targeting both the ER pathway and the cyclin D-CDK4 / 6-INK4-Rb pathway usually results in more It is inhibited extensively, the activation of escape pathways is prevented, and the development of endocrine therapy resistance is hindered. See Sammons et al., Current Cancer Drug Targets 17:637-649, 2017.

[0136] Thus, in certain embodiments, the breast cancer is pRb+ breast cancer. In certain embodiments, the breast cancer is hormone receptor (HR) positive (e.g., estrogen receptor positive (ER + ), progesterone receptor positive (PR + ), or ER + PR + ), HR + HER2 - or ER + HER2 - and is a HER2 / neu negative cancer, a progressive or metastatic or recurrent breast cancer. In certain embodiments, the HR + HER2 - or ER + HER2 - progressive or metastatic or recurrent breast cancer is present in adult women, or postmenopausal women.

[0137] In certain embodiments, the compounds of the invention are used alone or in combination with an aromatase inhibitor (which inhibits estrogen production) to treat HR positive, HER2 negative progressive or metastatic or recurrent breast cancer. In certain embodiments, the aromatase inhibitor temporarily inactivates aromatase (such as anastrozole (ARIMIDEX® and letrozole (FEMARA® etc.). In certain embodiments, the aromatase inhibitor permanently inactivates aromatase (such as exemestane (AROMASIN® etc.).

[0138] In certain embodiments, the compounds of the invention are used with compounds that interfere with the ability of estrogen to stimulate the growth of breast cancer cells, such as selective estrogen receptor modulators (SERMs) that bind to estrogen receptors to prevent estrogen binding, such as tamoxifen (NOLVADEX®) and toremifene (FARESTON®). Tamoxifen has been used in the treatment of HR + breast cancer for over 30 years.

[0139] In certain embodiments, the compounds of the invention are used with pure anti-estrogens that do not have estrogen agonist activity, such as fulvestrant (FASLODEX®).

[0140] In certain embodiments, HR-positive, HER2-negative advanced or metastatic or recurrent breast cancer is present in postmenopausal women. In certain embodiments, HR-positive, HER2-negative advanced or metastatic or recurrent breast cancer has progressed after treatment that changes the patient's hormones (e.g., estrogen and / or progesterone), or has worsened after treatment with another hormonal therapy.

[0141] In certain embodiments, the compounds of the invention are used in patients who have undergone or have had oophorectomy. In certain embodiments, oophorectomy is by ovariectomy or radiation therapy.

[0142] In certain embodiments, the compounds of the invention are used with compounds that temporarily suppress ovarian function (e.g., estrogen and / or progesterone production). Such compounds include gonadotropin-releasing hormone (GnRH) agonists or luteinizing hormone-releasing hormone (LH-RH) agonists, including goserelin (ZOLADEX®) and leuprolide (LUPRON®).

[0143] In certain embodiments, the compounds of the present invention are used with compounds that inhibit CYP3A4, such as ritonavir, indinavir, nelfinavir, saquinavir, clarithromycin, telithromycin, chloramphenicol, ketoconazole, itraconazole, posaconazole, voriconazole, nefazodone, cobicistat, amiodarone, aprepitant, verapamil, diltiazem, erythromycin, fluconazole, miconazole, bergamottin, cimetidine, ciprofloxacin, cyclosporine, dronedarone, fluvoxamine, imatinib, valerian, buprenorphine, caffeic acid, cilostazol, fosaprepitant, gabapentin, lomitapide, orphenadrine, ranitidine, ranolazine, tacrolimus, ticagrelor, valproic acid, amlodipine, cannabidiol, dithiocarbamate, mifepristone, norfloxacin, delavirdine, gestodene, mibefradil, starfruit, milk thistle, niacinamide, ginkgo biloba, piperine, isoniazid, and quercetin. In certain embodiments, the compounds of the present invention are used with inhibitors of IGF-1 / IGF-2, such as monoclonal antibodies against IGF-1 / IGF-2 or antigen-binding fragments thereof. An exemplary antibody is xentuzumab, a humanized IgG1 mAb.

[0144] In certain embodiments, the compounds of the present invention are used with compounds that inhibit PI3K. Inhibition of PI3K is thought to reduce the levels of cyclin D1 and other G1-S cyclins, abrogate pRb phosphorylation, and inhibit the activation of the S-phase transcriptional program. Representative PI3K inhibitors for use with the compounds of the present invention include idelalisib, copanlisib, duvelisib, taselisib, perifosine, buparlisib, alpelisib, umbralisib, copanlisib, ductilisib, and bosterlisib.

[0145]

[0146] ​In certain embodiments, the mammal to be treated is a human such as an adult female with breast cancer (e.g., a postmenopausal or adult female having hormone receptor (HR)-positive, human epidermal growth factor receptor 2 (HER2)-negative advanced or metastatic or recurrent breast cancer that progressed after receiving treatment that changed the patient's hormones).

[0147] Furthermore, certain compounds of the present invention exhibit the advantageous property that they can cross the blood-brain barrier. Thus, such compounds can penetrate the brain and are therefore useful for the treatment of primary and metastatic brain tumors in which cells are proliferating and contain a functional and intact Rb1 gene. Such pRb + Examples of brain tumors include glioblastoma, as well as medulloblastoma and astrocytoma (Lee et al., Science 235:1394, 1987).

[0148] Temozolomide is a cytotoxic DNA alkylating agent used for the treatment of brain tumors including glioblastoma and astrocytoma, including brain metastases from melanoma, breast cancer, and NSCLC (Friedman et al., Clin. Cancer Res. 6(7):2585-2597, 2000) (Siena et al., Annals of Oncology, doi:10.1093 / annonc / mdp343, 2009). Temozolomide interacts with DNA and causes chemical modification / damage (Marchesi et al., Pharmacol. Res. 56(4):275-287, 2007). Thus, in some embodiments, the compounds of the present invention are for the treatment of primary and metastatic pRb such as glioblastoma and astrocytoma + For the treatment of brain tumors, for example, such metastases can be used in combination with temozolomide when such metastases are derived from melanoma, breast cancer, or NSCLC.

[0149] 5. Pharmaceutical Compositions The present invention provides a pharmaceutical composition comprising any one of the compounds described herein, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers or excipients.

[0150] "Pharmaceutically acceptable excipient" and "pharmaceutically acceptable carrier" refer to substances that assist in the formulation and / or administration of the active agent to the subject and / or absorption by the subject, and mean substances that can be included in the compositions of the present disclosure without causing harmful toxicological effects to the subject. Non-limiting examples of pharmaceutically acceptable carriers and excipients include water, NaCl, physiological saline, lactated Ringer's solution, ordinary sucrose, ordinary glucose, binders, fillers, disintegrants, lubricants, coating agents, sweeteners, flavoring agents, salt solutions (such as Ringer's solution), alcohols, oils, gelatin, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidone, and coloring agents. Such preparations can be sterilized and, if necessary, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifying agents, salts for affecting osmotic pressure, buffers, coloring agents, and / or flavoring agents that do not react detrimentally with the compounds provided herein or interfere with their activity. Those skilled in the art will recognize that other pharmaceutical carriers and excipients are suitable for use with the disclosed compounds.

[0151] These compositions may optionally further comprise one or more additional therapeutic agents. Alternatively, the compounds of the present invention can be administered to patients in need thereof in combination with the administration of one or more other therapeutic regimens (e.g., Gleevec or other kinase inhibitors, interferon, bone marrow transplantation, farnesyltransferase inhibitors, bisphosphonates, thalidomide, cancer vaccines, hormone therapy, antibodies, radiation, etc.). For example, the additional therapeutic agent for combination with or inclusion in the pharmaceutical composition comprising the compound of the present invention can be one or more other anti-cancer agents.

[0152] As described herein, the compositions of the present invention, as used herein, include the compounds of the present invention together with a pharmaceutically acceptable carrier, which includes any and all solvents, diluents, or other vehicles, dispersing or suspending aids, surfactants, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants, etc. suitable for the particular dosage form desired. Remington’s Pharmaceutical Sciences, Fifteenth Edition, E.W. Martin (Mack Publishing Co., Easton, Pa., 1975) discloses various carriers used in the formulation of pharmaceutical compositions and known techniques for their preparation. For example, the use thereof is contemplated to be within the scope of the present invention, except when any conventional carrier medium is incompatible with the compounds of the present invention by producing undesirable biological effects or otherwise interacting in a detrimental manner with any other component of the pharmaceutical composition. Some examples of materials that can serve as pharmaceutically acceptable carriers include sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives (e.g., sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate); powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil; corn oil, and soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate. Similarly, coloring agents, release agents, coating agents, sweetening agents, flavoring agents, and fragrances, preservatives, and antioxidants may also be present in the composition.

[0153] 6. Formulation The present invention also encompasses a class of compositions comprising the active compounds of the invention, together with one or more pharmaceutically acceptable carriers and / or diluents and / or adjuvants (collectively referred to herein as "carrier" materials), and, optionally, other active ingredients, as appropriate.

[0154] In certain embodiments, the present invention provides a pharmaceutical formulation for treating cancer, particularly cancer as described herein, comprising a compound of the invention or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier.

[0155] In certain embodiments, the present invention provides a pharmaceutical formulation for treating cancer selected from the group consisting of colorectal cancer, mantle cell lymphoma, breast cancer (including ER + HER2 - progressive or metastatic or recurrent breast cancer in adult or postmenopausal women), glioblastoma, acute myeloid leukemia, and lung cancer, particularly NSCLC, comprising a compound of the invention or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier.

[0156] In certain embodiments, the present invention provides a pharmaceutical formulation for treating glioblastoma or astrocytoma, comprising a compound of the invention and temozolomide, together with a pharmaceutically acceptable carrier.

[0157] In certain embodiments, the present invention also provides a pharmaceutical formulation comprising a compound of the invention or a pharmaceutically acceptable salt thereof and temozolomide, together with a pharmaceutically acceptable carrier, diluent, or excipient.

[0158] In certain embodiments, the present invention provides a pharmaceutical formulation for treating NSCLC, pancreatic cancer, ovarian cancer, or metastatic breast cancer (including ER + HER2 - progressive or metastatic or recurrent breast cancer in adult or postmenopausal women), comprising the present invention and gemcitabine HCl, together with a pharmaceutically acceptable carrier.

[0159] In certain embodiments, the present invention also provides a pharmaceutical formulation comprising a compound of the present invention or a pharmaceutically acceptable salt thereof and gemcitabine HCl, together with a pharmaceutically acceptable carrier, diluent, or excipient.

[0160] The active compounds of the present invention are preferably administered by such a route in the form of a pharmaceutical composition adapted to any suitable route, in an amount effective for the intended treatment. The compounds and compositions of the present invention are in dosage unit formulations containing conventional pharmaceutically acceptable carriers, adjuvants, and vehicles, and can be administered, for example, orally, mucosally, topically, rectally, by inhalation spray, etc., for pulmonary or parenteral (including intravascular, intravenous, intraperitoneal, subcutaneous, intramuscular, intrasternal, and infusion techniques) administration.

[0161] The pharmaceutically active compounds of the present invention can be processed according to conventional pharmacy methods to produce agents for administration to patients, including humans and other mammals.

[0162] For oral administration, the pharmaceutical composition can be, for example, in the form of tablets, capsules, suspensions, or liquids. The pharmaceutical composition is preferably prepared in the form of dosage units containing a specific amount of the active ingredient.

[0163] Examples of such dosage units are tablets or capsules. For example, the suitable daily dosage for a human or other mammal can vary depending on the condition of the patient and other factors, but can also be determined using routine methods in this case.

[0164] The amount and dosing schedule of the compounds of the present invention and / or the compounds administered to treat a medical condition depend on various factors including the age, weight, sex, and medical condition of the subject, the type of disease, the severity of the disease, the route and frequency of administration, and the specific compound used. Accordingly, the dosing schedule can vary widely, but a physician can use standard methods It can be determined daily. As described above, the daily dose can be administered in a single dose or divided into two, three, four or more doses.

[0165] For therapeutic purposes, the active compounds of the invention are usually combined with one or more adjuvants, excipients or carriers suitable for the indicated route of administration. When administered orally, the compounds are mixed with lactose, sucrose, starch powder, cellulose esters of alkanoic acids, cellulose alkyl esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids, gelatin, acacia gum, sodium alginate, polyvinylpyrrolidone, and / or polyvinyl alcohol and then can be tableted or encapsulated as convenient for administration. Such capsules or tablets can contain sustained-release formulations such as can be provided as a dispersion of the active compound in hydroxypropylmethylcellulose.

[0166] In the case of skin conditions, it may be preferable to apply the topical preparation of the compounds of the invention to the affected area 2 to 4 times a day. Formulations suitable for topical administration include liquid or semi-liquid preparations suitable for penetration into the skin (e.g., liniments, lotions, ointments, creams, or pastes) and drops suitable for administration to the eyes, ears, or nose. For topical administration, the active ingredient can contain from 0.001% to 10% w / w, for example, from 1% to 2% by weight of the formulation, and may contain 10% w / w, but is preferably 5% w / w or less, more preferably from 0.1% to 1%.

[0167] The compounds of the present invention can also be administered by a transdermal device. Preferably, transdermal administration is achieved using a patch of either the reservoir and porous membrane type or the solid matrix type. In either case, the active agent is delivered from the reservoir or microcapsules continuously through the membrane to an adhesive part permeable to the active agent in contact with the recipient's skin or mucosa. When the active agent is absorbed through the skin, a controlled and predetermined flow rate of the active agent is administered to the recipient. In the case of microcapsules, the encapsulating agent can also function as a membrane. The oil phase of the emulsion of the present invention can be composed of known components by known methods.

[0168] The phase may contain only an emulsifier, but may also contain a mixture of at least one emulsifier and a fat or an oil, or a mixture of both a fat and an oil. Preferably, the hydrophilic emulsifier is included together with a lipophilic emulsifier that acts as a stabilizer. It is also preferred to include both an oil and a fat. Together, with or without a stabilizer, the emulsifiers constitute a so-called emulsifying wax, which together with the oil and fat constitutes a so-called emulsifying ointment base that forms the oily dispersed phase of the cream preparation. Emulsifiers and emulsion stabilizers suitable for use in the formulations of the present invention include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate, sodium lauryl sulfate, glyceryl distearate alone or containing a wax, or other materials well known in the art.

[0169] Since the solubility of the active compound in most oils that are likely to be used in pharmaceutical emulsion formulations is very low, the selection of a suitable oil or fat for the formulation is based on achieving the desired appearance characteristics. Thus, the cream should preferably be a non-greasy, non-staining, washable product with a suitable consistency to avoid leakage from tubes or other containers. Straight-chain or branched-chain, mono- or dibasic alkyl esters, such as diisoadipates, isocetyl stearate, propylene glycol diesters of coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate, or blends of branched-chain esters may be used. These can be used alone or in combination depending on the required properties. They can also be used in combination.

[0170] Alternatively, high melting point lipids such as white soft paraffin and / or liquid paraffin or other mineral oils can be used.

[0171] Formulations suitable for topical administration to the eye also include eye drops in which the active ingredient is dissolved or suspended in a suitable carrier, particularly an aqueous solvent for the active ingredient.

[0172] The active ingredient is preferably present in such formulations at a concentration of 0.5 - 20%, advantageously 0.5 - 10%, particularly about 1.5% w / w.

[0173] Formulations for parenteral administration can be in the form of aqueous or non-aqueous isotonic sterile injection solutions or suspensions. These solutions and suspensions can be prepared from sterile powders or granules using one or more of the carriers or diluents mentioned for use in formulations for oral administration, or by using other suitable dispersing or wetting agents and suspending agents. These compounds can be dissolved in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, tragacanth gum, and / or various buffer solutions. Other adjuvants and modes of administration are well and widely known in the pharmaceutical field. The active ingredient can also be administered by injection as a composition comprising a suitable carrier containing physiological saline, dextrose, or water, or cyclodextrin (i.e., Captisol), cosolvent solubilization (i.e., propylene glycol) or micelle solubilization (i.e., Tween80).

[0174] Sterile injectable preparations can also be in the form of sterile injection solutions or suspensions in a non-toxic parenterally acceptable diluent or solvent, such as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be used are water, Ringer's solution, and isotonic sodium chloride solution. Furthermore, sterile, fixed oils have conventionally been used as a solvent or suspending medium. For this purpose, any bland fixed oil containing synthetic monoglycerides or diglycerides can be used. In addition, fatty acids such as oleic acid have been used in the preparation of injectables.

[0175] In the case of pulmonary administration, the pharmaceutical composition can be administered in the form of an aerosol or using an inhaler containing a dry powder aerosol.

[0176] Suppositories for rectal administration of drugs can be prepared by mixing the drug with suitable non-irritating excipients such as cocoa butter and polyethylene glycol, which are solid at normal temperature but liquid at rectal temperature and thus melt in the rectum to release the drug.

[0177] The pharmaceutical composition can be subjected to conventional pharmaceutical processes such as sterilization and / or can contain conventional adjuvants such as preservatives, stabilizers, wetting agents, emulsifiers, buffer solutions, etc. Tablets and pills can be further prepared with enteric coatings. Such compositions can also contain auxiliary agents such as wetting agents, sweeteners, flavoring agents, and fragrances. The pharmaceutical composition of the present invention comprises a compound of the formula described herein or a pharmaceutically acceptable salt thereof; an additional agent selected from a kinase inhibitor (small molecule, polypeptide, antibody, etc.), an immunosuppressant, an anticancer agent, an antiviral agent, an anti-inflammatory agent, an antifungal agent, an antibiotic, or an anti-angiogenic compound; and any pharmaceutically acceptable carrier, adjuvant or vehicle.

[0178] An alternative composition of the present invention comprises a compound of the formula described herein or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, adjuvant or vehicle. Such a composition can optionally contain one or more additional therapeutic agents including, for example, a kinase inhibitor (small molecule, polypeptide, antibody, etc.), an immunosuppressant, an anticancer agent, an antiviral agent, an anti-inflammatory agent, an antifungal agent, an antibiotic, or an anti-angiogenic compound.

[0179] The term "pharmaceutically acceptable carrier or adjuvant" refers to a carrier or adjuvant that can be administered to a patient together with a compound of the present invention, is non-toxic when administered in a dosage sufficient to deliver a therapeutic amount of the compound without destroying its pharmacological activity. Pharmaceutically acceptable carriers, adjuvants, or vehicles that can be used in the pharmaceutical compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS) such as D-α-tocopherol polyethylene glycol 1000 succinate, surfactants used in pharmaceutical dosage forms, such as Tween or other similar polymer delivery matrices, serum albumin such as human serum albumin, buffering substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and lanolin. Cyclodextrins such as u-, P-, and y-cyclodextrin, or chemically modified derivatives such as hydroxyalkyl cyclodextrins containing 2 and 3-hydroxypropyl-cyclodextrin, or other solubilizing derivatives can also be advantageously used to enhance the delivery of the compounds of the formulas described herein.

[0180] The pharmaceutical composition can be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, emulsions and aqueous suspensions, dispersions and solutions. In the case of oral tablets, commonly used carriers include lactose and corn starch. Lubricants such as magnesium stearate are also typically added. In the case of oral administration in capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions and / or emulsions are orally administered, the active ingredient can be suspended or dissolved in the oil phase and combined with emulsifying and / or suspending agents.

[0181] Optionally, certain sweetening, flavoring and / or coloring agents can be added. The pharmaceutical composition can include formulations utilizing liposome or microencapsulation technology, examples of which are known in the art.

[0182] The pharmaceutical composition can be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the art of pharmaceutical formulations and can be prepared as solutions in physiological saline using benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and / or other solubilizing or dispersing agents, examples of which are well known in the art.

[0183] 7. Therapeutic Kit One aspect of the present invention relates to a kit for conveniently and effectively carrying out a method or use according to the present invention. Generally, a pharmaceutical pack or kit includes one or more containers filled with one or more components of the pharmaceutical composition of the present invention. Such kits are particularly suitable for the delivery of solid oral dosage forms such as tablets or capsules. Such kits preferably include several unit doses and may also include a card having the doses oriented in the order of their intended use. Optionally, a memory aid may be provided, for example, in the form of numbers, letters, or other marks, or together with a calendar insert specifying the dates of the treatment schedule on which the doses may be administered. Such containers may optionally be accompanied by a notice in a form prescribed by a government agency that regulates the manufacture, use, or sale of the pharmaceutical, the notice reflecting the agency's approval of the manufacture, use, or sale for human administration.

[0184] The following representative examples include important additional information, illustrations, and guidance that may be adapted for the practice of the invention in various embodiments and their equivalents. The following examples are intended to assist in illustrating the invention and are not intended, nor should they be construed, to limit the scope of the invention. Indeed, various modifications of the invention, and many additional embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art upon reviewing this document, including references to the following examples, as well as scientific and patent documents cited herein.

[0185] The content of the cited references is incorporated herein by reference to help explain the prior art.

[0186] Furthermore, for the purposes of the present invention, chemical elements are in the CAS version, Handbook of Chemistry and Physics, 75 thIt is identified according to the periodic table of the elements on the front and back covers of the Ed. Further, the general principles of organic chemistry, as well as specific functional moieties and reactivities, are described in "Organic Chemistry," Thomas Sorrell, University Science Books, Sausalito: 1999, and "Organic Chemistry," Morrison & Boyd (3d Ed), the entire contents of both of which are incorporated herein by reference.

[0187] 8. Synthetic Scheme The compounds of formula I can be prepared by one of ordinary skill in the art according to techniques and procedures recognized in the art. More specifically, the compounds of formula I can be prepared as described in the schemes, methods, and examples set forth below. It will be recognized by one of ordinary skill in the art that the individual steps of the following schemes can be varied to provide the compounds of formula I. Reagents and starting materials are readily available to one of ordinary skill in the art. Unless otherwise specified, all substituents are as previously defined.

Examples

[0188] Biological Example 1. CDK4 / Cyclin D1 Inhibition Assay IC 50The CDK4 enzyme assay for determination was performed as follows. Using the microfluidic kinase detection technology (Caliper), phosphorylation of the peptide substrate by CDK4 / cyclin D1 was monitored. The total reaction volume was 15 μL containing buffer A (100 mM HEPES (pH 7.5), 0.1% BSA, 0.01% Triton X-100, 1 mM DTT, 10 mM MgCl2, 10 μM sodium orthovanadate, 10 μM beta-glycerophosphate), 200 μM ATP, 1 nM CDK4 / cyclin D1 (Thermofisher, PR8064A), 1 μM FL-34 (5-FAM-RRRFRPASPLRGPPK), and the test compound appropriately diluted with DMSO. All components were added to a 384-well plate (Corning, 4514) and incubated at room temperature for 3 hours. The reaction was stopped by the addition of 15 μL of stop buffer (180 mM HEPES (pH 7.5), 20 mM EDTA, Coating-3 reagent (PerkinElmer, 760050)). The plate was then loaded into a Caliper EZ Reader (EZ Reader II, PerkinElmer, HD-4HYSG2772), and the reaction mixture containing the substrate and product was introduced into the microfluidic chip for separation and detection. The IC 50 value of the test compound was determined by fitting the inhibition curve to a four-parameter sigmoid dose-response model using Xlfit5 / GraphPad Prism5 software.

[0189] Biological Example 2. Assay for Inhibition of CDK6 / Cyclin D3 IC 50The CDK6 enzyme assay for determination was performed as follows. Using a microfluidic kinase detection technology (Caliper), phosphorylation of a peptide substrate by CDK6 / cyclin D3 was monitored. The total reaction volume was 15 μL containing buffer A (100 mM HEPES (pH 7.5), 0.1% BSA, 0.01% Triton X-100, 1 mM DTT, 10 mM MgCl2, 10 μM sodium orthovanadate, 10 μM beta-glycerophosphate), 300 μM ATP, 2 nM CDK6 / cyclin D3 (Carna, 04-107), 1 μM FL-34 (5-FAM-RRRFRPASPLRGPPK), and a test compound appropriately diluted with DMSO. All components were added to a 384-well plate (Corning, 4514) and incubated at room temperature for 3 hours. The reaction was stopped by the addition of 15 μL of stop buffer (180 mM HEPES (pH 7.5), 20 mM EDTA, Coating-3 reagent (PerkinElmer, 760050)). The plate was then loaded onto a Caliper EZ Reader (EZ Reader II, PerkinElmer, HD-4HYSG2772), and the reaction mixture containing the substrate and product was introduced into the microfluidic chip for separation and detection. The IC 50 value was determined by fitting the inhibition curve with a four-parameter sigmoid dose-response model using Xlfit5 / GraphPad Prism5 software.

[0190] Biological Example 3. Inhibition Assay of CDK2 / Cyclin E1 IC 50The CDK2 enzyme assay for determination was carried out as follows. Using the microfluidic kinase detection technology (Caliper), the phosphorylation of the peptide substrate by CDK2 / cyclin E1 was monitored. The total reaction volume was 15 μL containing buffer A (100 mM HEPES (pH 7.5), 0.1% BSA, 0.01% Triton X-100, 1 mM DTT, 10 mM MgCl2, 10 μM sodium orthovanadate, 10 μM beta-glycerophosphate), 100 μM ATP, 5 nM CDK2 / cyclin E1 (SignalChem, C29-18G), 5 μM FL-18 (5-FAM-QSPKKG-NH2), and the test compound appropriately diluted with DMSO. All components were added to a 384-well plate (Corning, 4514) and incubated at room temperature for 3 hours. The reaction was stopped by the addition of 15 μL of stop buffer (180 mM HEPES (pH 7.5), 20 mM EDTA, Coating-3 reagent (PerkinElmer, 760050)). The plate was loaded into a Caliper EZ Reader (EZ Reader II, PerkinElmer, HD-4HYSG2772), and the reaction mixture containing the substrate and product was introduced into the microfluidic chip for separation and detection. The IC 50 value of the test compound was determined by fitting the inhibition curve with a four-parameter sigmoid dose-response model using Xlfit5 / GraphPad Prism5 software.

[0191] The IC 50 value of each exemplary compound against CDK2, CDK4, and CDK6 is provided in the synthesis examples described below. The IC 50 values are shown as "A", "B", "C", and "D" for values of 10 nM or less, 100 nM or less, 1 μM or less, and greater than 1 μM, respectively.

[0192] Biological Example 4. Anti-proliferation assay in T47D cells T47D is a human breast cancer cell line commonly used in biomedical research related to the hormonal expression of cancer cells. T47D cells are different from other human breast cancer cells in that the progesterone receptor (PR) is not regulated by estradiol, a hormone that is abundant in the cells themselves. T47D cells have been used in the study of the effects of progesterone on breast cancer and the corresponding transcriptional regulation induced by introduced drugs. This cell has been found to be highly resistant to estrogen and anti-estrogens.

[0193] T47D breast cancer cells (ATCC, HTB-133) from the American Type Culture Collection were seeded at 3000 cells / well in 96-well plates and incubated at 37 °C, 5% CO2 in RPMI1640 medium (Gibco, 31800105) containing 10% fetal bovine serum (FBS, Biowest, FB-1058). After an overnight incubation, the baseline value of the samples from one plate was measured using the Cyquant reagent (Invitrogen, C35011) according to the manufacturer's recommendation. After incubating the cells with the detection reagent for 1 hour at 37 °C, fluorescence was measured at 485 nm excitation and 535 nm emission using a Spectra Max M5 (Molecular Devices, HD-4HYSG3196). Compounds were added to other plates at 10 dose concentrations from 10 μM to 0.51 nM in a 3-fold dilution scheme. On the 6th day after compound addition, the Cyquant reagent was added and fluorescence was measured using a Spectra Max M5. The IC 50 value of the test compound's anti-proliferative activity was determined from the survival rate reading curve subtracted from the baseline using Xlfit5 / GraphPad Prism5 software.

[0194] Biological Example 5. Inhibition of phosphorylation of retinoblastoma protein (pRb) in T47D cells T47D breast cancer cells (ATCC, HTB-133) from the American Type Culture Collection were seeded in 96-well plates at 40,000 cells / well and incubated in RPMI 1640 medium (Gibco, 31800105) containing 10% fetal bovine serum (FBS, Biowest, FB-1058). The cells were then allowed to adhere overnight at 37 °C and 5% CO2. The next day, when the compounds were titrated in a 3-fold dilution scheme, the highest compound concentration tested was 10 μM. After 24 hours of incubation with the compounds, the cells were lysed in ice-cold lysis buffer containing a phosphatase inhibitor cocktail and 1 mM PMSF. The cell lysates (50 μL / well) were then transferred to ELISA plates (pRb Ser807 / 811 ELISA kit, Cell Signaling, 13152 or pRb Ser780 ELISA kit, Cell Signaling, 13016). The plates were incubated overnight at 4 °C with constant gentle shaking. After incubation, the plates were washed according to the manufacturer's recommendations, and then 100 μL of reconstituted detection antibody was added to each well and incubated at 37 °C for 1 hour. After incubation, the plates were washed, and then 100 μL of reconstituted HRP-conjugated secondary antibody was added to each well and incubated at 37 °C for 30 minutes. After incubation, the plates were washed. Then, 100 μL of TMB substrate was added to each well and incubated at 37 °C for 10 minutes or at 25 °C for 30 minutes. Finally, 100 μL of STOP solution was added to each well and gently mixed for a few seconds. The plates were read using an Envision plate reader (PerkinElmer, 2104-0010) in 96-well luminescence mode. IC 50 values were calculated using a 4-parameter sigmoid dose-response model of Xlfit5 / GraphPad Prism5 software.

[0195] The cell data obtained from Biological Examples 4 and 5 are listed in Table A below. IC 50The values are shown as "++++" for values of 100 nM or less, "+++" for values of 500 nM or less, "++" for values of 1 μM or less, and "+" for values exceeding 1 μM, respectively.

[0196] Synthesis Example Description of Equipment 1 The 1H NMR spectrum was recorded on a Bruker Ascend 400 spectrometer. Chemical shifts are expressed in parts per million (ppm, δ units). Coupling constants are in the unit of Hertz (Hz). The splitting pattern represents the apparent multiplicity and is denoted as s (singlet), d (doublet), t (triplet), q (quartet), quint (quintet), m (multiplet), br (broad). doublet).

[0197] The low-resolution mass spectrum (MS) for analysis was recorded on an ACQUITY UPLC manufactured by Waters equipped with an SQ Detector, using a CORTECS C18 +, 2.7 μm 4.6 × 30 mm column manufactured by Waters, with gradient elution method.

[0198] Solvent A: 0.1% formic acid (FA) in water Solvent B: 0.1% FA in acetonitrile From 5% ACN to 95% ACN in 1.0 minute, held for 1.0 minute, Total 2.5 minutes, flow rate: 1.8 mL / min, column temperature 40 degrees.

[0199] Intermediate Intermediate 1

Chemical Structure

[0200] Step 1 To a solution of 4-benzyloxypyridine (185 mg, 998 μmol) in DCM (10 mL), amino 2,4,6-trimethylbenzenesulfonate (236 mg, 1.1 mmol) was added at 25 °C. The reaction mixture was stirred at 25 °C for 14 h. The mixture was concentrated under reduced pressure to afford the desired crude product (400 mg, 99% yield) as a colorless oil. LC-MS: m / z 202 [M+H] + .

[0201] Step 2 To a solution of 4-benzyloxypyridin-1-ium-1-amine (187 mg, 929 μmol) in DMF (10 mL), Cs2CO3 (192 mg, 1.4 mmol) and but-3-yn-2-one (94 mg, 1.4 mmol) were added. The reaction mixture was stirred at 25 °C for 12 h. The reaction mixture was quenched with water (50 mL) and extracted with DCM (2 × 25 mL). The organic layer was washed with brine (20 mL), dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by column chromatography (eluting with PE / EA = 10 / 1) to afford the desired product (90 mg, 35% yield) as a yellow solid. LC-MS: m / z 267 [M+H] + .

[0202] Step 3 To a solution of methyl(triphenyl)phosphonium bromide (241 mg, 675 μmol) in THF (10 ml), butyllithium (43.3 mg, 675 μmol) was added dropwise at -20 °C under N2 and the reaction was stirred at -20 °C for 1 h. Then, a solution of 1-(5-benzyloxypyrazolo[1,5-a]pyridin-3-yl)ethanone (90 mg, 338 μmol) in THF (15 ml) was added dropwise at -20 °C. The reaction mixture was stirred at 10 °C for 3 h. The reaction mixture was quenched with MeOH (3 ml) and concentrated under reduced pressure. The residue was purified by preparative HPLC (eluting with PE:EA = 1 / 1) to afford the desired crude product (41.0 mg, 46% yield) as a yellow solid. LC-MS: m / z 265 [M+H] + .

[0203] Step 4 To a solution of 5-benzyloxy-3-isopropenyl-pyrazolo[1,5-a]pyridine (600 mg, 2.3 mmol) in methanol (50 mL) was added Pd / C (60 mg). The reaction mixture was stirred at 30 °C for 48 h under H2. The reaction mixture was filtered and concentrated under reduced pressure to afford the desired product (380 mg, 95% yield) as a yellow solid. LC-MS: m / z 177 [M+H] + 。

[0204] Step 5 To a solution of 3-isopropylpyrazolo[1,5-a]pyridin-5-ol (650 mg, 3.7 mmol) and DIPEA (410 mg, 4.1 mmol) in DCM (15 mL) was added Tf2O (1.1 g, 4.1 mmol) at 0 °C under N2. The reaction mixture was stirred at 0 °C for 2 h. The reaction mixture was washed with brine (15 mL) and dried over Na2SO4. The organic layer was filtered and the filtrate was concentrated to afford the desired product (1.1 g, 92% yield) as a colorless oil. LC-MS: m / z 309 [M+H] + 。

[0205] Step 6 To a solution of (3-isopropylpyrazolo[1,5-a]pyridin-5-yl)trifluoromethanesulfonate (1.1 g, 3.4 mmol) and 4,4,5,5-tetramethyl-2-(4,4,5,5)-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (1.3 g, 5.1 mmol) in dioxane (10 mL) were added Pd(dppf)Cl2 (249 mg, 340 μmol) and KOAc (1.0 g, 10.2 mmol). The reaction mixture was stirred at 110 °C for 2 h under N2. The mixture was filtered and the filtrate was concentrated under reduced pressure to afford the desired crude product (950 mg, 97% yield) as a dark solid. LC-MS: m / z 287 [M+H] + 。

[0206] Step 7 A solution of 3-isopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo[1,5-a]pyridine (950 mg, 3.3 mmol) and 2,4-dichloro-5-fluoro-pyrimidine (665 mg, 4.0 mmol) in H2O (1 mL) and 1,4-dioxane (15 mL) was added with Na2CO3 (1.2 g, 10.0 mmol) and Pd(dppf)Cl2 (242 mg, 332 μmol). The mixture was stirred at 110 °C for 6 h under N2. The mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography (20 g silica gel column, EA 0 - 50% in PE) to afford the desired product (650 mg, yield 67%) as a yellow solid. LC-MS: m / z 291 [M+H] + 。

[0207] Intermediate 2

Chemical Structure

[0208] To a solution of 3-isopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo[1,5-a]pyridine (3.5 g, 12.2 mmol) and 2,4-dichloropyrimidine (2.7 g, 18.4 mmol) in water (3 mL) and 1,4-dioxane (60 mL) were added Pd(dppf)Cl2 (0.9 g, 1.2 mmol) and Na2CO3 (1.52 g, 14 mmol). The reaction mixture was stirred at 110 °C for 6 h under N2. The mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography (20 g silica gel column, PE containing EA 0 - 50%) to afford the desired product (2.1 g, yield 62%) as a yellow solid. LC-MS: m / z 273 [M+H] + 。

[0209] Additional intermediates of the present invention were prepared by using the corresponding derivatives. The selected compounds and their corresponding characteristic data are shown in the following table.

Table 1

[0210] Intermediate 5 [Chem.]

[0211] 3-Isopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo [1,5-a]pyridine (200 mg, 698 μmol) and 2-chloro-5-fluoro-4-iodo-pyridine (269 mg, 1.1 mmol) in H2O (1 mL) and 1,4-dioxane (20 mL), Na2CO3 (260 mg, 2.1 mmol) and Pd(dppf)Cl2 (51.1 mg, 69.9 μmol) were added. The reaction mixture was stirred at 110 °C for 6 h under N2. The mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography (20 g silica gel column, 0 - 50% EA in PE) to give the desired product (140 mg, 69% yield) as a yellow solid. LC-MS: m / z 290 [M+H] + .

[0212] Intermediate 6 [Chem.]

[0213] Step 1 To a solution of 5-bromopyrazolo[1,5-a]pyridine (0.9 g, 4.6 mmol) in dry dioxane (40 mL), B2Pin2 (1.8 g, 6.9 mmol), Pd(dppf)Cl2 (0.7 g, 0.9 mmol) and potassium acetate (1.4 g, 13.9 mmol) were added. The mixture was stirred at 110 °C for 8 h in a nitrogen atmosphere. The reaction mixture was filtered and concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (0 - 50% petroleum ether / EtOAc) to give the desired product (1.1 g, 75% yield) as a white solid. LC-MS: m / z 245 [M+H] + .

[0214] Step 2 To a solution of 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo[1,5-a]pyridine (1.0 g, 4.0 mmol) in dioxane (45 mL) were added 2,4-dichloro-5-fluoropyrimidine (1.0 g, 6.0 mmol), Pd(dppf)Cl2 (0.6 g, 0.8 mmol) and K2CO3 (1.7 g, 12.0 mmol). Then 5 mL of H2O was added. The mixture was stirred at 110 °C for 8 h. The mixture was concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (0 - 50% EtOAc in petroleum ether) to give the desired product (0.7 g, 66% yield) as a white solid. LC-MS: m / z 249 [M+H] + .

[0215] Step 3 To a solution of 5-(2-chloro-5-fluoropyrimidin-4-yl)pyrazolo[1,5-a]pyridine (610 mg, 2.5 mmol) in DCM (20 mL) was added NBS (482 mg, 2.7 mmol). The mixture was stirred at 25 °C for 4 h. The mixture was concentrated under reduced pressure and purified by silica gel chromatography (0 - 50% EtOAc / PE) to give the desired product (680 mg, 84% yield) as a white solid. LC-MS: m / z 327 [M+H] + .

[0216] Intermediate 7 [Chemical formula]

[0217] Step 1 A solution of 5-bromo-3-iodo-pyrazolo[1,5-a]pyridine (1.0 g, 3.1 mmol) and cyclopent-1-ylboronic acid (0.4 g, 3.4 mmol) in dioxane (20 mL) and water (5 mL) was added with cyclopentyl(diphenyl)phosphane dichloromethane dichloropalladium iron (0.8 g, 0.9 mmol) and tripotassium phosphate (2.0 g, 9.3 mmol). The reaction mixture was stirred at 100 °C for 5 h under a N2 atmosphere. The reaction mixture was extracted with EtOAc (3 × 10 mL). The organic phase was washed with brine (50 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography to obtain the desired product (0.3 g, 38% yield) as a white solid. LC-MS: m / z 265.1 [M+H] + 。

[0218] Step 2 To a solution of 5-bromo-3-(cyclopent-1-yl)pyrazolo[1,5-a]pyridine (270 mg, 1.0 mmol) in methanol (10 mL) was added PtO2 (46.6 mg, 205.2 μmol). The mixture was stirred at 25 °C for 2 h under a hydrogen atmosphere. The mixture was filtered through a Celite pad. The filtrate was concentrated to obtain the desired product (190 mg, 69% yield) as a white solid, which was used directly in the next step without further purification. LC-MS: m / z 267.1 [M+H] + 。

[0219] Step 3 To a stirred solution of 5-bromo-3-cyclopentyl-pyrazolo[1,5-a]pyridine (210 mg, 792 μmol) and 4,4,5,5-tetramethyl-2-(4,4,5,5)-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (301 mg, 1.2 mmol) at 25 °C was added cyclopentyl(diphenyl)phosphane dichloropalladium iron (57.9 mg, 79.2 μmol) and potassium acetate (233 mg, 2.4 mmol). The reaction mixture was stirred at 110 °C for 2 h. The mixture was filtered and concentrated in vacuo to give the desired product as a black solid, which was used directly in the next step without further purification. LC-MS: m / z 313.1 [M+H] + 。

[0220] Step 4 To a mixture of 3-cyclopentyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo[1,5-a] pyridine (210 mg, 672 μmol) and 2,4-dichloro-5-fluoro-pyrimidine (134 mg, 807 μmol) in dioxane (5 mL) and water (1 mL) was added cyclopentyl(diphenyl)phosphane dichloropalladium iron (49.2 mg, 67.2 μmol), potassium carbonate (278 mg, 2.0 mmol). The resulting mixture was stirred at 105 °C for 3 h under N2 atmosphere. The reaction mixture was diluted with water (10 mL) and then extracted with EtOAc (3 × 10 mL). The combined organic phases were washed with brine (50 mL), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography to give the desired product (40 mg, yield 19%) as a white solid. LC-MS: m / z 317.1 [M+H] + 。

[0221] Intermediate 8

Chemical formula

[0222] Step 1 A solution of 5-bromo-3-iodo-pyrazolo[1,5-a]pyridine (1.0 g, 3.1 mmol) and 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (845.7 mg, 4.0 mmol) in anhydrous dioxane (20 mL) was added with Pd(dppf)Cl2 (758 mg, 929 μmol), K3PO4 (2.0 g, 9.3 mmol) and water (5 mL) under a N2 atmosphere. Then, the reaction mixture was stirred at 110 °C for 1 h. The mixture was concentrated under reduced pressure, and the residue was purified by flash column chromatography (80 g silica gel column, petroleum ether / EtOAc with 0 - 30% EtOAc) to obtain the desired product (508 mg, yield 59%) as a brown solid. LC-MS: m / z 279 [M+H] + 。

[0223] Step 2 To a solution of 5-bromo-3-(3,6-dihydro-2H-pyran-4-yl)pyrazolo[1,5-a]pyridine (360 mg, 1.3 mmol) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (425 mg, 1.7 mmol) in dioxane (15 mL) were added cyclopentyl(diphenyl)phosphane dichloropalladium iron (188 mg, 257 μmol) and potassium acetate (379 mg, 3.8 mmol) under a N2 atmosphere. Then, the reaction mixture was stirred at 110 °C for 8 h. The mixture was filtered and concentrated under reduced pressure to obtain the desired crude product as a yellow oil, which was used in the next step without further purification. LC-MS: m / z 327 [M+H] + 。

[0224] Step 3 3-(3,6-Dihydro-2H-pyran-4-yl)-5 in dioxane (15 mL) To a solution of (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo[1,5-a]pyridine (430 mg, 1.3 mmol) and 2,4-dichloro-5-fluoro-pyrimidine (242 mg, 1.5 mmol) were added cyclopentyl(diphenyl)phosphane dichloropalladium iron (192 mg, 263 μmol), sodium carbonate (419 mg, 3.9 mmol) and water (1 mL) under N2 atmosphere. The reaction mixture was then stirred at 110 °C for 3 h. The mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography (40 g silica gel column, 0 - 30% EtOAC in petroleum ether) to afford the desired product (360 mg, 82% yield) as an orange solid. LC-MS: m / z 331.1 [M+H] + 。

[0225] Intermediate 9

Chemical formula

[0226] Step 1 To a solution of 5-(2-chloropyrimidin-4-yl)-3-isopropyl-pyrazolo[1,5-a]pyridine (100 mg, 366 μmol) in THF (8 mL) was added TMPMgCl-LiCl (1 M, 916 μL) at -78 °C under N2. The reaction was stirred at -78 °C for 0.5 h. Then NBS (78.3 mg, 439 μmol) was added. The mixture was warmed to 25 °C and stirred for 1.5 h. The mixture was quenched with aqueous NH4Cl solution (10 mL) and extracted with EA (2×15 mL). The combined organic phases were concentrated under reduced pressure. The residue was purified by FCC (12 g silica gel, 0 - 30% EtOAc in PE) to afford the desired product (45.0 mg, 34% yield) as a yellow solid. LC-MS: m / z 351.0 [M+H] + 。

[0227] Step 2 A solution of 7-bromo-5-(2-chloropyrimidin-4-yl)-3-isopropyl-pyrazolo[1,5-a]pyridine (45.0 mg, 127 μmol), Pd(PPh3)4 (14.7 mg, 12.8 μmol), and Zn(CN)2 (22.5 mg, 191 μmol) in NMP (5 mL) was irradiated in a microwave reactor at 110 °C for 0.5 h under N2. The mixture was concentrated under reduced pressure. The residue was purified by FCC (4 g silica gel, 0 - 30% EtOAc in PE) to give the desired product (15.0 mg, 39% yield) as a yellow solid. LC-MS: m / z 298.1 [M+H] + 。

[0228] Intermediate 84

Chemical Structure

[0229] Step 1 To a suspension of aluminum trichloride (2.7 g, 20.3 mmol) in DCM (10 mL) was added 2-bromo-2-methyl-propane (4.1 g, 30.4 mmol) at 0 °C under a N2 atmosphere. The reaction mixture was stirred at 0 °C for 10 min under a N2 atmosphere. Then, 5-bromopyrazolo[1,5-a]pyridine (2 g, 10.1 mmol) was added to the reaction mixture. The reaction mixture was stirred at 0 °C for 3 h. The reaction mixture was quenched with ice water (50 mL). Then, the mixture was extracted with DCM (3 × 50 mL), and the combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash column chromatography to give the desired product, 5-bromo-3-tert-butyl-pyrazolo[1,5-a]pyridine (1.2 g, 46% yield), as a yellow oil. LC-MS: m / z 253.1 [M+H] + 。

[0230] Step 2 A solution of 5-bromo-3-tert-butyl-pyrazolo[1,5-a]pyridine (200 mg, 790 μmol) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (300 mg, 1.2 mmol) in dioxane (8 mL) was added with cyclopentyl(diphenyl)phosphane dichloropalladium iron (173 mg, 237 μmol) and potassium acetate (232 mg, 2.4 mmol) under a N2 atmosphere. Then, the mixture was stirred at 110 °C for 13 h under a N2 atmosphere. The mixture was filtered and concentrated under reduced pressure to obtain the desired product as a yellow oil, which was used in the next step without further purification.

[0231] Step 3 To a solution of (3-tert-butylpyrazolo[1,5-a]pyridin-5-yl)boronic acid (180 mg, 825 μmol) and 2,4-dichloropyrimidine (147 mg, 990 μmol) in dioxane (12 mL) were added cyclopentyl(diphenyl)phosphane dichloropalladium iron (72 mg, 99 μmol), sodium carbonate (174 mg, 1.6 mmol) and water (0.5 mL) under a N2 atmosphere. Then, the reaction mixture was stirred at 110 °C for 5 h under a N2 atmosphere. The mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography to obtain the desired product (220 mg, 92% yield) as a yellow solid. LC-MS: m / z 287.1 [M+H] + 。

[0232] Additional intermediates of the present invention were prepared by using the corresponding derivatives. The selected compounds and their corresponding characteristic data are shown in the following table.

Table 2

[0233] Intermediate 10

Chemical formula

[0234] Step 1 To a solution of 5-bromo-3-iodo-2H-pyrazolo[3,4-c]pyridine (200 mg, 617 μmol) and Cs2CO3 (500 mg, 1.5 mmol) in DMF (8 mL) was added MeI (99 mg, 679 μmol) at 0 °C. The reaction mixture was stirred at 25 °C for 1.5 h. The mixture was quenched with water (20 mL) and then extracted with ethyl acetate (2 × 50 mL). The combined organic phases were dried over anhydrous Na2SO4. The filtrate was concentrated. The residue was purified by flash column chromatography (24 g silica gel column, 0 - 10% EA in DCM in 20 min) to give the desired product (20.0 mg) as a pale purple solid. LC-MS: m / z 337.9 [M+H] + 。

[0235] Step 2 A solution of 5-bromo-3-iodo-2-methyl-pyrazolo[3,4-c]pyridine (167 mg, 494 μmol), potassium isopropenyltrifluoroborate (90.0 mg, 600 μmol), Pd(dppf)Cl2 (90.0 mg, 100 μmol) and K2CO3 (140 mg, 1 mmol) in dioxane (10 mL) was stirred at 80 °C for 72 h. The reaction mixture was quenched with water (50 mL) and then extracted with ethyl acetate (2 × 100 mL). The combined organic phases were dried over anhydrous Na2SO4 and then filtered. The filtrate was concentrated. The residue was purified by flash column chromatography (24 g silica gel column, 0 - 70% EA in PE in 20 min) to give the desired product (30.0 mg) as a yellow solid. LC-MS: m / z 252.1 [M+H] + 。

[0236] Step 3 A solution of 5-bromo-3-isopropenyl-2-methyl-pyrazolo[3,4-c]pyridine (40.0 mg, 158 μmol) in THF (12 mL) was added with PtO2 (18.1 mg, 79.3 μmol), and the mixture was stirred at 25 °C for 2 h under a hydrogen atmosphere. The reaction mixture was filtered and then washed with methanol (5 mL). The filtrate was concentrated to give the desired product (30.0 mg, 74% yield) as a white solid, which was used directly without further purification. LC-MS: m / z 254.1 [M+H] + .

[0237] Step 4 A solution of 5-bromo-3-isopropyl-2-methyl-pyrazolo[3,4-c]pyridine (30.0 mg, 118 μmol), B2Pin2 (46.0 mg, 181 μmol), Pd2(dba)3 (22.0 mg, 24.0 μmol), KOAc (35.0 mg, 357 μmol) and tricyclohexylphosphane (14.0 mg, 0.05 mmol) in dioxane (2 mL) was stirred at 120 °C for 1 h under microwave conditions. The reaction mixture was filtered and washed with DCM (5 mL). The filtrate was concentrated under reduced pressure to give the desired crude product (25.0 mg) as a black solid. LC-MS: m / z 220.1 [M+H] + .

[0238] Step 5 A solution of (3-isopropyl-2-methyl-pyrazolo[3,4-c]pyridin-5-yl)boronic acid (26.0 mg, 118 μmol), 2,4-dichloro-5-fluoro-pyrimidine (24.0 mg, 14 μmol), Pd(dppf)Cl2 (9.0 mg, 12.3 μmol) and K2CO3 (33.0 mg, 239 μmol) in dioxane (3 mL) and water (1.5 mL) was stirred at 100 °C for 16 h under a N2 atmosphere. The reaction mixture was quenched with water (50 mL) and then extracted with ethyl acetate (2 × 100 mL). The combined organic phases were dried over anhydrous Na2SO4 and then filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (24 g, 0 - 90% EA in PE for 20 min) to give the desired product (30.0 mg) as a yellow solid. LC-MS: m / z 306.1 [M+H] + 。

[0239] Intermediate 11

Chem.

[0240] Step 1 Iron powder (2.6 g, 46.5 mmol), water (9 mL), and concentrated HCl (2 mL) were added to 4-amino-2,6-dichloro-3-nitropyridine (2.0 g, 9.6 mmol) in ethanol (50 mL). The mixture was heated to reflux for 16 h. The mixture was cooled to room temperature and then neutralized with sodium hydrogen carbonate (saturated aqueous solution). The mixture was filtered and the residue was washed with ethyl acetate. The filtrate was concentrated. The residue was dissolved in ethyl acetate and washed with water (30 mL). The organic layer was dried over Na2SO4 and evaporated to give the desired product (1.9 g, 99% yield) as a yellow solid. LC-MS: m / z 177.9 [M+H] + 。

[0241] Step 2 A solution of 2,6-dichloropyridine-3,4-diamine (1.8 g, 8.4 mmol) in trimethyl orthoacetate (20 mL) was stirred at 140 °C for 5 h. The mixture was then concentrated under reduced pressure and the residue was dissolved in AcOH (20 mL). The mixture was then stirred at 120 °C for 5 h. The mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography (40 g silica gel column, 0 - 50% EtOAc in petroleum ether) to give the desired product (1.3 g, 67% yield) as a yellow solid. LC-MS: m / z 201.9 [M+H] + 。

[0242] Step 3 To a solution of 4,6-dichloro-2-methyl-1H-imidazo[4,5-c]pyridine (1.9 g) in anhydrous DMF (5 mL) was added NaH (772 mg, 32.1 mmol) at 0 °C . Then, 2-iodopropane (3.3 g, 19.3 mmol) was added to the reaction mixture. The mixture was warmed to room temperature and stirred for 13 h. The mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (80 g silica gel column, 0 - 10% MeOH in DCM) to give the desired product (487 mg, 31% yield) as a yellow solid. LC-MS: m / z 244 [M+H] + 。

[0243] Step 4 To a solution of 4,6-dichloro-1-isopropyl-2-methyl-1H-imidazo[4,5-c]pyridine (200 mg, 0.8 mmol) in MeOH (5 mL) was added a solution of MeONa (5.4 N, 5 mL in MeOH). The mixture was heated to reflux for 10 h (65 °C). The mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (40 g silica gel column, 0 - 100% EtOAc in petroleum ether) to give the desired product (140 mg, 71% yield) as a white solid. LC-MS: m / z 240 [M+H] + 。

[0244] Step 5 A solution of 6-chloro-1-isopropyl-4-methoxy-2-methyl-1H-imidazo[4,5-c]pyridine (120 mg, 0.5 mmol) and Pin2B2 (153 mg, 0.6 mmol) in anhydrous dioxane (8 mL) was added with Pd2(dba)3 (137 mg, 0.15 mmol), tricyclohexylphosphine (84.1 mg, 0.3 mmol), and AcOK (147 mg, 1.5 mmol) under a N2 atmosphere. Subsequently, the mixture was stirred at 110 °C (microwave) for 1.5 h. The mixture was filtered and concentrated under reduced pressure to obtain the desired crude product as a brown oil, which was used in the next step without further purification. LC-MS: m / z 250 [M+H] + 。

[0245] Step 6 A solution of (1-isopropyl-4-methoxy-2-methyl-1H-imidazo[4,5-c]pyridin-6-yl)boronic acid (300 mg) and 2,4-dichloro-5-fluoropyrimidine (83.5 mg, 0.5 mmol) in anhydrous dioxane / water (8 mL / 2 mL) was added with Pd(dppf)Cl2 (122 mg, 0.2 mmol) and Na2CO3 (159 mg, 1.5 mmol) under a N2 atmosphere. Subsequently, the mixture was stirred at 110 °C for 3 h. The mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (40 g silica gel column, 0 - 50% EtOAc in petroleum ether) to obtain the desired product (139 mg, yield 82%) as a yellow solid. LC-MS: m / z 336 [M+H] + 。

[0246] Additional intermediates of the present invention were prepared by using the corresponding derivatives. The selected compounds and their corresponding characteristic data are shown in the following table.

Table 3

[0247] Intermediate 18

Chemical formula

[0248] Step 1 A solution of 6-chloro-1-isopropyl-4-methoxy-2-methyl-imidazo[4,5-c]pyridine (170 mg, 709 μmol) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (270 mg, 1.1 mmol) in dioxane (5 mL) was placed in a sealed tube. Then, tris(dibenzylideneacetone)dipalladium(0) (194 mg, 212 μmol), tricyclohexylphosphane (119 mg, 425 μmol) and potassium acetate (208 mg, 2.1 mmol) were added under a N2 atmosphere. The reaction mixture was then stirred at 110 °C for 5 h. The mixture was filtered and the filtrate was concentrated under reduced pressure to obtain the desired crude product, which was used in the next step without further purification. LC-MS: m / z 250 [M+H] 。 + 。

[0249] Step 2 To a solution of (1-isopropyl-4-methoxy-2-methyl-imidazo[4,5-c]pyridin-6-yl)boronic acid (190 mg, 762 μmol) and 2-chloro-5-fluoro-4-iodo-pyridine (216 mg, 839 μmol) in dioxane (8 mL), cyclopentyl(diphenyl)phosphane dichloropalladium iron (167 mg, 228 μmol), disodium carbonate (242 mg, 2.2 mmol) and water (2 mL) were added under a N2 atmosphere. The reaction mixture was then stirred at 110 °C for 5 h. The mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography (40 g silica gel column, petroleum ether / EtOAc with EtOAc 0 - 100%) to obtain the desired product (160 mg, yield 62%) as a yellow solid. LC-MS: (ESI) m / z 335 [M+H] + 。

[0250] Intermediate 19

Chemical Structure

[0251] Step 1 To a mixture of 4,6-dichloro-1-isopropyl-imidazo[4,5-c]pyridine (250 mg, 1.1 mmol) in DMSO (10 mL) was added CsF (510 mg, 3.4 mmol), and then the mixture was stirred at 140 °C for 1.5 h. The resulting mixture was poured into water (100 mL) and extracted with EA (3 × 30 mL). The combined organic layers were dried over Na2SO4 and filtered. The filtrate was concentrated. The residue was purified by flash column (80 g, 200-300 mesh silica gel, PE / EA = 5 / 1-2 / 1) to give the desired product (210 mg, 75% yield) as a milky white solid. LC-MS: (ESI) m / z 214.1 [M+H].

[0252] Step 2 6-Chloro-4-fluoro-1-isopropyl-imidazo[4,5-c]pyridine (30.0 mg, 140 μmol) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl) in dioxane (5 mL). To a solution of cyclopentyl(diphenyl)phosphane-1,3,2-dioxaborolane (35.6 mg, 140 μmol) was added potassium acetate (41.3 mg, 421 μmol) and cyclopentyl(diphenyl)phosphane dichloropalladium iron (15.4 mg, 21.1 μmol). The mixture was degassed with N2 and stirred at 110 °C for 16 h. The mixture was filtered through a Celite pad. The filtrate was concentrated under reduced pressure to give the desired crude product (50 mg) as a black oil, which was used directly in the next step. LC-MS: (ESI) m / z 224.2 [M+H] + .

[0253] Step 3 To a solution of (4-fluoro-1-isopropyl-imidazo[4,5-c]pyridin-6-yl)boronic acid (50.0 mg, 224 μmol) and 2-chloro-4-iodo-pyrimidine (53.9 mg, 224 μmol) in dioxane (3 mL) was added cyclopentyl(diphenyl)phosphane dichloropalladium iron (24.6 mg, 33.6 μmol) and potassium acetate (66.0 mg, 672 μmol). The mixture was degassed with N2 and stirred at 110 °C for 16 h. The mixture was concentrated under reduced pressure. The residue was purified by flash chromatography eluting with 0 - 60% ethyl acetate in petroleum ether to afford the desired product (30.0 mg, 46% yield) as a white solid. LC-MS: (ESI) m / z) 292.1 [M+H] + 。

[0254] Additional intermediates of the present invention were prepared by using the corresponding derivatives. The selected compounds and their corresponding characteristic data are shown in the following table.

Table 4

[0255] Intermediate 20

Chem.

[0256] Step 1 To a solution of 4,6-dichloro-2-methyl-3H-imidazo[4,5-c]pyridine (4.0 g, 19.8 mmol) and 6-oxabicyclo[3.1.0]hexane (6.6 g, 79.1 mmol) in DMF (50 mL) was added cesium carbonate (16.1 g, 49.5 mmol) and the reaction mixture was stirred at 100 °C for 48 h. The reaction mixture was then quenched with H2O (50 mL) and extracted with EA (3 × 10 mL). The organic layer was concentrated under reduced pressure. The residue was purified by flash column chromatography (20 g silica gel column, DCM containing 0 - 10% MeOH) to afford the desired product (650 mg, 11% yield) as a yellow solid. LC-MS: m / z 286 [M+H]+ .

[0257] Step 2 To a solution of 2-(4,6-dichloro-2-methyl-imidazo[4,5-c]pyridin-1-yl)cyclopentanol (650 mg, 2.2 mmol) in methanol (10 mL) was added sodium methoxide (5.4 M, 841 μL)) at 25 °C. The reaction mixture was stirred at 60 °C for 48 h. The mixture was quenched with water (20 mL) and then extracted with ethyl acetate (2 × 20 mL). The combined organic phases were dried over sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with 10 - 60% EA in PE to give the desired product (600 mg, 94% yield) as a pale yellow oil. LC-MS: (ESI) m / z 282.2 [M+H] + .

[0258] Step 3 To a mixture of 2-(6-chloro-4-methoxy-2-methyl-imidazo[4,5-c]pyridin-1-yl)cyclopentanol (200 mg, 709 μmol) and 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (270 mg, 1.1 mmol) in dioxane (8 mL) were added Pd2(dba)3 (97.5 mg, 106 μmol), potassium acetate (209 mg, 2.1 mmol) and tricyclohexylphosphane (59.7 mg, 212 μmol). The resulting mixture was stirred at 110 °C for 6 h under a nitrogen atmosphere. The mixture was filtered and the filtrate was concentrated under reduced pressure to give the desired crude product (200 mg, 96% yield) as a brown oil, which was used in the next step without further purification. LC-MS: (ESI) m / z 292.2 [M+H] + .

[0259] Step 4 A solution of [1-(2-Hydroxycyclopentyl)-4-methoxy-imidazo[4,5-c]pyridin-6-yl]boronic acid (250 mg, 902 μmol) and 2,4-dichloro-5-fluoro-pyrimidine (165 mg, 992 μmol) in dioxane (3 mL) was added with sodium carbonate (239 mg, 2.3 mmol) and cyclopentyl(diphenyl)phosphane dichloropalladium(II) (132 mg, 180 μmol) under a N2 atmosphere. The reaction mixture was then stirred at 110 °C for 5 h. The mixture was concentrated under reduced pressure and the residue was purified to give the desired product (250 mg, 76% yield) as a yellow solid. LC-MS: m / z 378.1 [M+H] + 。

[0260] Additional intermediates of the present invention were prepared by using the corresponding derivatives. The selected compounds and their corresponding characteristic data are shown in the following table.

Table 5

[0261] Intermediate 22

Chem.

[0262] Step 1 To a solution of 6-(2-Chloropyrimidin-4-yl)-1-isopropyl-4-methoxy-imidazo[4,5-c]pyridine (150 mg, 493 μmol) in DCE (5 mL) was added tribromoborane (618 mg, 2.4 mmol) at 0 °C. The mixture was stirred at 60 °C for 12 h. The mixture was quenched with aqueous NaHCO3 (10 mL) and extracted with DCM (2 × 10 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated to give the desired crude product (140 mg) as a yellow solid, which was used directly in the next step without further purification. LC-MS: m / z 290.1 [M+H] + 。

[0263] Step 2 A solution of 6-(2-chloropyrimidin-4-yl)-1-isopropyl-imidazo[4,5-c]pyridin-4-ol (0.1 g, 345 μmol) and trimethylsilyl 2,2-difluoro-2-fluorosulfonyl-acetate (129 mg, 517 μmol) in CH3CN (5 mL) was added with NaH (16.5 mg, 690 μmol) and CsF (78.6 mg, 517 μmol) at 0 °C. The mixture was stirred at 0 °C for 1 h. The mixture was quenched with water (5 mL) and extracted with EtOAc (2 × 10 mL). The combined organic layers were dried over Na2SO4, filtered, concentrated to give a residue, which was purified by flash column chromatography (SiO2, hexane / ethyl acetate 1:1) to afford the desired product (105 mg, 89% yield) as a white solid. LC-MS: m / z 340.1 [M+H] + .

[0264] Intermediate 23 [Chemical Structure]

[0265] Step 1 To a mixture of tert-butyl 2-chloro-7,8-dihydro-5H-1,6-naphthyridine-6-carboxylate (302 mg, 2.3 mmol) in dioxane (10 mL), 4-(1-isopropyl-4-methoxy-imidazo[4,5-c]pyridin-6-yl)pyrimidin-2-amine (322 mg, 1.1 mmol), tris(dibenzylideneacetone)dipalladium(0) (100 mg, 109 μmol), (5-diphenylphosphanyl-9,9-dimethyl-xanthen-4-yl)-diphenyl-phosphane (190 mg, 328 μmol), and cesium carbonate (2.0 g, 6.1 mmol) were added in sequence. The mixture was degassed 5 times with N2. The mixture was stirred at 110 °C for 16 h. The reaction mixture was diluted with water (100 mL) and extracted with EA (3 × 30 mL). The combined organic layers were dried over Na2SO4, filtered, and the filtrate was concentrated. The residue was purified by flash column chromatography to afford the desired product (611 mg, 92% yield) as a yellow solid. LC-MS: (ESI) m / z 517.3 [M+H] + 。

[0266] Step 2 In an ice bath, HCl / EtOAc (2 M, 7 mL) was added to tert-butyl 2-[[4-(1-isopropyl-4-methoxy-1H-imidazo[4,5-c]pyridin-6-yl)pyrimidin-2-yl]amino]-7,8-dihydro-5H-1,6-naphthyridine-6-carboxylate (152 mg, 280 μmol), and the mixture was stirred in the ice bath for 2 h. The mixture was concentrated and dried in vacuo to afford the desired product (122 mg, 91% yield) as a yellow solid. LC-MS: (ESI) m / z 417.2 [M+H] + 。

[0267] Intermediate 24

Chemical Structure

[0268] Step 1 To a solution of 2-bromo-6-fluorophenol (4.0 g, 21.1 mmol) in DMF (50 mL) were added K2CO3 (5.8 g, 42.2 mmol), TBAI (0.4 g, 1.1 mmol), and 3-chloro-2-methylprop-1-ene (2.8 g, 31.6 mmol) at 0 °C. The mixture was stirred at 25 °C for 14 h. The mixture was concentrated under reduced pressure and purified to afford the desired product (4.5 g, 88% yield). LC-MS: m / z 245.0 [M+H] + 。

[0269] Step 2 To a solution of 1-bromo-3-fluoro-2-((2-methylallyl)oxy)benzene (2.5 g, 10.2 mmol) in toluene (50 mL) were added n-Bu3SnH (3.6 g, 12.3 mmol) and AIBN (2.0 g, 2.3 mmol) under N2. The reaction mixture was stirred at 90 °C for 12 h. The reaction mixture was quenched with water (50 mL) and extracted with DCM (2 × 50 mL). The organic layer was washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by column chromatography (eluting with PE / EA = 10 / 1) to afford the desired product (1.4 g, 81% yield). LC-MS: m / z 167.1 [M+H] + 。

[0270] Step 3 To a solution of 7-fluoro-3,3-dimethyl-2,3-dihydrobenzofuran (1.4 g, 8.4 mmol) in DMF (30 mL) was added NBS (1.8 g, 10.1 mmol) under N2 at 25 °C. The reaction was stirred for 15 h. The reaction mixture was quenched with water (3 mL) and concentrated under reduced pressure. The residue was purified (eluting with PE:EA = 1 / 1) to afford the desired product (450 mg, 22% yield). LC-MS: m / z 245.0 [M+H] + 。

[0271] Step 4 A solution of 5-bromo-7-fluoro-3,3-dimethyl-2,3-dihydrobenzofuran (100 mg, 0.4 mmol) and Pin2B2 (122 mg, 0.5 mmol) in anhydrous dioxane (10 mL) was added with Pd(dppf)Cl2 (98.0 mg, 0.16 mmol) and KOAc (117 mg, 1.2 mmol) under a N2 atmosphere. Then, the mixture was stirred at 110 °C (microwave) for 1.5 h. The mixture was filtered and concentrated under reduced pressure to obtain the desired crude product (150 mg) as a brown oil, which was used in the next step without further purification. LC-MS: m / z 293.2 [M+H] + 。

[0272] Step 5 To a solution of 2-(7-fluoro-3,3-dimethyl- 2,3-dihydrobenzofuran-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (150 mg) and 2,4-dichloropyrimidine (61.0 mg, 0.4 mmol) in anhydrous dioxane / water (8 mL / 2 mL) were added Pd(dppf)Cl2 (98.0 mg, 0.16 mmol), Na2CO3 (127 mg, 1.2 mmol) under a N2 atmosphere. Then the mixture was stirred at 110 °C for 3 h. The mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (40 g silica gel column, EtOAc in petroleum ether / EtOAc 0 - 50%) to obtain the desired product (60.0 mg, yield 51%). LC-MS: m / z 279.1 [M+H] + 。

[0273] Intermediate 25

Chemical Structure

[0274] Step 1 To a mixture of 4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine (2.3 g, 17.3 mmol) and DIPEA (4.4 g, 34 mmol) in methanol (30 mL) was added Boc2O (5.5 g, 43.2 mmol) at 25 °C. The reaction mixture was stirred at 25 °C for 18 h. The reaction mixture was quenched with water (50 mL) and then extracted with EtOAc (2 × 100 mL). The combined organic phases were dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated. The residue was purified by flash column chromatography (24 g, 0 - 100% EtOAc in PE for 20 min) to afford the desired product (4.7 g, 14.5 mmol) as a yellow oil. LC-MS: m / z 346.2 [M+Na] + .

[0275] Step 2 A mixture of di-tert-butyl 6,7-dihydro-4H-imidazo[4,5-c]pyridine-1,5-dicarboxylate (4.7 g, 14.5 mmol) and sodium hydroxide (1 M in water, 29.0 mL) in dioxane (36 mL) was stirred at 25 °C for 1 h. The reaction mixture was quenched with water (50 mL) and then extracted with EtOAc (2 × 100 mL). The combined organic phases were dried over anhydrous Na2SO4 and then filtered. The filtrate was concentrated. The residue was purified by flash column chromatography to afford the desired product (2.7 g, 12.1 mmol) as a colorless oil. LC-MS: m / z 224.3 [M+H] + .

[0276] Step 3 A mixture of tert-butyl 1,4,6,7-tetrahydroimidazo[4,5-c]pyridine-5-carboxylate (500 mg, 2.2 mmol) and cesium carbonate (2.1 g, 6.7 mmol) in DMF (20 mL) was stirred at 25 °C for 16 h under N2 atmosphere. The reaction mixture was quenched with water (50 mL) and then extracted with EtOAc (2 × 100 mL). The combined organic phases were dried over anhydrous Na2SO4 and then filtered. The filtrate was It was concentrated. The residue was purified by flash column chromatography (24 g, MeOH 0 - 20% in DCM for 20 min) to give the desired product (330 mg, 55% yield) as a yellow oil. LC-MS: m / z 266.2 [M+H] + .

[0277] Step 4 A mixture of tert-butyl 3-isopropyl-6,7-dihydro-4H-imidazo[4,5-c]pyridine-5-carboxylate (530 mg, 2.0 mmol) and HCl (2N, 2 mL) in ethyl acetate (2 mL) was stirred at 25 °C for 1 h. The reaction mixture was quenched with water (50 mL) and then extracted with EtOAc (2 × 100 mL). The combined organic phases were dried over anhydrous Na2SO4 and then filtered. The filtrate was concentrated. The residue was purified by flash column chromatography (24 g, EtOAc 0 - 100% in PE for 20 min) to give the desired product (300 mg, 90% yield) as a yellow solid. LC-MS: m / z 166.3 [M+H] + .

[0278] Step 5 A mixture of 3-isopropyl-4,5,6,7-tetrahydroimidazo[4,5-c]pyridine (165 mg, 998 μmol), 2,4-dichloropyrimidine (178 mg, 1.2 mmol) and potassium carbonate (690 mg, 4.9 mmol) in methanol (10 mL) was stirred at 40 °C for 2 h under N2 atmosphere. The reaction mixture was quenched with water (50 mL) and then extracted with EtOAc (2 × 100 mL). The combined organic phases were dried over anhydrous Na2SO4 and then filtered. The filtrate was concentrated. The residue was purified by flash column chromatography (24 g, MeOH 0 - 20% in DCM for 20 min) to give the desired product (66.0 mg) as a colorless oil. LC-MS: m / z 278.1 [M+H] + .

[0279] Additional intermediates of the present invention were prepared by using the corresponding derivatives. The selected compounds and their corresponding characteristic data are shown in the following table.

Table 6

[0280] Intermediate 26

Chem.

[0281] Step 1 To a solution of 5-bromo-2-nitro-pyridine (10.0 g, 49.2 mmol) and tert-butyl 3-oxopiperazine-1-carboxylate (10.8 g, 54.1 mmol) in dioxane (30 mL) was added cesium carbonate (32.1 g, 98.5 mmol ), tris(dibenzylideneacetone)dipalladium(0) (4.5 g, 4.9 mmol) and (5-diphenylphosphanyl-9,9-dimethyl-xanthen-4-yl)-diphenyl-phosphane (2.8 g, 4.9 mmol) at 25 °C. The mixture was then stirred at 110 °C for 8 h. The solvent was removed under reduced pressure. The residue was purified by silica gel chromatography eluting with 0 - 15% MeOH in DCM over 20 min to give the desired product (1.5 g, 9% yield) as a pale yellow solid. LC-MS: m / z 323.1 [M+H] + .

[0282] Step 2 To a stirred solution of tert-butyl 4-(6-nitro-3-pyridyl)-3-oxo-piperazine-1-carboxylate (1.6 g, 4.9 mmol) in ethanol (20 mL) was added iron powder (1.1 g, 19.8 mmol) and ammonium chloride (2.6 g, 49.6 mmol) at 25 °C. The reaction mixture was stirred at 80 °C for 2 h under a N2 atmosphere. The mixture was filtered through a Celite pad. The filtrate was concentrated under reduced pressure to give the desired product, which was used directly in the next step without further purification. LC-MS: m / z 293.1 [M+H] + .

[0283] By using the corresponding derivatives, additional intermediates of the present invention were prepared. The selected compounds and their corresponding characteristic data are shown in the following table.

Table 7-1

Table 7-2

Table 7-3

Table 7-4

[0284] Intermediate 40

Chem.

[0285] Step 1 To a solution of 2-methylmorpholine (100 mg, 988 μmol) and 5-fluoro-2-nitro-pyridine (140 mg, 988 μmol) in ethanol (15 mL) was added DIPEA (383 mg, 2.9 mmol). The mixture was stirred at 80 °C for 8 hours. The reaction was concentrated under reduced pressure to give the desired product (200 mg, 90% yield) as a yellow solid. LC-MS: m / z 224.1 [M+H] + 。

[0286] Step 2 To a solution of 2-methyl-4-(6-nitro-3-pyridyl)morpholine (200 mg, 895 μmol) in ethanol (15 mL) was added iron (250 mg, 4.4 mmol) and ammonium chloride (239 mg, 4.4 mmol). The mixture was stirred at 80 °C for 8 h. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with MeOH / DCM 0 - 10% in 15 min to afford the desired product (150 mg, 86% yield) as a brown solid. LC-MS: m / z 194.1 [M+H] + 。

[0287] Intermediate 115

Chemical formula

[0288] Step 1 To a solution of 5-bromo-2-nitro-pyridine (1 g, 4.9 mmol) and 1-isopropylpiperazine (631.6 mg, 4.9 mmol) in dioxane (40 mL) were added tris(dibenzylideneacetone)dipalladium(0) (451.1 mg, 492 μmol), (5-diphenylphosphanyl-9,9-dimethyl-xanthen-4-yl)-diphenylphosphane (570 mg, 985 μmol) and cesium carbonate (4.8 g, 14.8 mmol). The reaction mixture was then stirred at 110 °C for 3 h under N2. The reaction mixture was concentrated under reduced pressure and purified by silica gel column chromatography eluting with ethyl acetate 1 - 100% in petroleum ether to afford the desired product (850 mg, 68% yield) as a yellow solid. LC-MS: m / z 251.1 [M+H] + 。

[0289] Step 2 To a solution of 1-isopropyl-4-(6-nitro-3-pyridyl)piperazine (850 mg, 3.4 mmol) in methanol (30 mL) was added Pd / C (412 mg, 10%). The reaction mixture was then degassed three times with H2 and stirred at 25 °C for 3 hours. The reaction mixture was filtered and then washed with methanol (20 mL). The combined solvents were concentrated under reduced pressure to give the desired product (620 mg, 82% yield) as a brown solid. LC-MS: m / z 221.2 [M+H] + 。

[0290] Additional intermediates of the present invention were prepared by using the corresponding derivatives. The selected compounds and their corresponding characteristic data are shown in the following table.

Table 8-1

Table 8-2

[0291] Intermediate 56

Chemical formula

[0292] Step 1 To a mixture of piperidine-2,4-dione (2.1 g, 18.5 mmol) and N-methylmethanamine (3.4 g, 74.2 mmol) in DCM (36 mL) and THF (18 mL), CH3COOH (10 mL) was added and the resulting mixture was stirred at 25 °C for 3 h under a nitrogen atmosphere. Sodium triacetoxyborohydride (7.8 g, 37.1 mmol) was added to this mixture and the resulting mixture was stirred at 25 °C for 12 h under a nitrogen atmosphere. The reaction was quenched with water (50 mL) and concentrated in vacuo to remove DCM and THF. The mixture was extracted with DCM (3 × 100 mL). The organic solution was washed with brine (20 mL). The organic phase was dried over Na2SO4, filtered, and concentrated under reduced pressure to give the desired product (2.6 g, 89% yield), which was used in the next step without purification. LC-MS: m / z 141.2 [M+H] + 。

[0293] Step 2 Sodium borohydride (539.0 mg, 14.2 mmol) was added to a mixture of 4-(dimethylamino)-2,3-dihydro-1H-pyridin-6-one (1.0 g, 7.1 mmol) in methanol (15 mL) and the resulting mixture was stirred at 25 °C for 12 h under a nitrogen atmosphere. The reaction was quenched with saturated aqueous NH4Cl solution (10 mL) and concentrated in vacuo to remove MeOH. The aqueous solution was purified by reverse-phase column (C18, 40 g) eluting with (MeCN / water (0.1% NH4OH) = 1 / 10) to give the desired product (0.3 g, 32% yield) as a pale yellow solid. LC-MS: m / z 143.2 [M+H] + 。

[0294] Step 3 A mixture of 4-(dimethylamino)piperidin-2-one (270 mg, 1.9 mmol), 5-iodopyridin-2-amine (1.0 g, 4.7 mmol) and potassium phosphate (1.2 g, 5.7 mmol) in dioxane (26 mL) was added to (1S,2S)-N1,N2-dimethylcyclohexane-1,2-diamine (162 mg, 1.1 mmol) and CuI (108 mg, 569 μmol). The resulting mixture was stirred at 110 °C for 12 h under a nitrogen atmosphere. The reaction mixture was filtered. The filtrate was concentrated in vacuo to give a residue. The residue was purified by a reverse-phase column (C18, 20 g) eluting with (MeCN / water (0.1% NH4OH) = 1 / 10) to give the desired product (272 mg, 61% yield) as a pale yellow solid. LC-MS: m / z 235.2 [M+H] + 。

[0295] Additional intermediates of the present invention were prepared by using the corresponding derivatives. The selected compounds and their corresponding characteristic data are shown in the following table.

Table 9-1

Table 9-2

[0296] Intermediate 135

Chemical formula

[0297] Step 1 To a solution of piperidine-2,4-dione (5 g, 44.2 mmol) in methanol (20 mL) was added methanamine (2.7 g, 88.4 mmol, 3.0 mL). The mixture was stirred at 25 °C for 16 h. The mixture was concentrated in vacuo to give the desired product as a brown solid. LC-MS: m / z 127.1 [M+1] + 。

[0298] Step 2 To a solution of 4-(methylamino)-2,3-dihydro-1H-pyridin-6-one (2.6 g, 20.8 mmol) in methanol (20 mL) was added dioxoplatinum (474.1 mg, 2.0 mmol). The mixture was then degassed with H2. The reaction mixture was stirred at 25 °C for 16 h. The mixture was filtered and concentrated in vacuo to afford the desired product as a black oil. LC-MS: m / z 129.1 [M+1] + .

[0299] Step 3 To a solution of 4-(methylamino)piperidin-2-one (2.6 g, 20.5 mmol) in DCM (10 mL) were added N,N-diethylethanamine (4.1 g, 41.1 mmol) and tert-butyl tert-butyl dicarbonate (5.3 g, 24.6 mmol). The reaction mixture was stirred at 25 °C for 3 h. The mixture was concentrated under reduced pressure and purified by flash chromatography (40 g silica gel, 0 - 10% MeOH in DCM) to afford the desired product (2.9 g, 61% yield) as a yellow oil. LC-MS: m / z 229.2 [M+1] + .

[0300] Step 4 To a solution of tert-butyl N-methyl-N-(2-oxo-4-piperidyl)carbamate (2 g, 8.7 mmol) in dry dioxane (20 mL) were added 5-iodopyridin-2-amine (1.9 g, 8.7 mmol), copper(I) iodide (166.8 mg, 876 μmol), (1R,2R)-N1,N2-dimethylcyclohexane-1,2-diamine (249 mg, 1.7 mmol) and tripotassium phosphate (5.5 g, 26.2 mmol). The reaction mixture was stirred at 105 °C for 16 h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (40 g silica gel, 0 - 10% MeOH in DCM) to afford the desired product (1.7 g, 62% yield) as a black oil. LC-MS: m / z 321.2 [M+1] + .

[0301] Additional intermediates of the present invention were prepared by using the corresponding derivatives. The selected compounds and their corresponding characteristic data are shown in the following table.

Table 10

[0302] Intermediate 61

Chem.

[0303] Step 1 In an ice bath, formic acid (610 mg, 13.2 mmol) and formaldehyde (540 mg, 17.9 mmol) were added to a mixture of 3-aminopyrrolidin-2-one (333 mg, 3.3 mmol) in water (8 mL). The resulting mixture was stirred at 100 °C for 1.5 h. The mixture was concentrated. The residue was purified by preparative HPLC and then lyophilized to give the desired product (162 mg) as a yellow oil. LC-MS: m / z 129.3 [M+H] + 。

[0304] Step 2 To a solution of 3-(dimethylamino)pyrrolidin-2-one (130 mg, 1.0 mmol) and 5-iodopyridine-2-amine (267 mg, 1.2 mmol) in dioxane (3 mL) were added (1S,2S)-N1,N2-dimethylcyclohexane-1,2-diamine (43 mg, 304 μmol), CuI (28.9 mg, 152 μmol) and tripotassium phosphate (645 mg, 3.0 mmol) at 25 °C. The reaction mixture was stirred at 110 °C for 16 h. The mixture was quenched with water (20 mL) and then extracted with ethyl acetate (2×10 mL). The combined organic phases were dried over sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with 20 - 70% EA in PE to give the desired product (160 mg, 71% yield) as a yellow solid. LC-MS: m / z 221.1 [M+H] + 。

[0305] By using the corresponding derivatives, additional intermediates of the present invention were prepared. The selected compounds and their corresponding characteristic data are shown in the following table. [Table 11]

[0306] Intermediate 62 [Chemical formula]

[0307] To a solution of 4-amino-1H-pyridin-2-one (4.0 g, 36.3 mmol) and tert-butyl 4-methylsulfonyloxypiperidine-1-carboxylate (10.1 g, 36.3 mmol) in DMF (10 mL) was added NaH (835 mg, 34.8 mmol) at 25 °C. The mixture was stirred at 45 °C for 6 hours. The reaction was quenched with water (200 mL) and then extracted with EA (3 × 100 mL). The organic solution was washed with brine (100 mL). The organic phase was dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with 0 - 15% methanol in dichloromethane in 20 minutes to give the desired product (0.4 g, 5% yield) as a yellow solid. LC-MS: m / z 294.2 [M+H] + 。

[0308] Intermediate 63 [Chemical formula]

[0309] Step 1 To a stirred solution of tert-butyl 3-oxo-1,4-diazepane-1-carboxylate (500 mg, 2.3 mmol) and 5-iodopyridin-2-amine (564 mg, 2.5 mmol) in dioxane (10 mL) were added (1R,2R)-N1,N2-dimethylcyclohexane-1,2-diamine (132 mg, 933 μmol), tripotassium phosphate (1.5 g, 7.0 mmol), and CuI (28.9 mg, 152 μmol). The reaction mixture was stirred at 110 °C for 5 h under a N2 atmosphere. The mixture was concentrated under reduced pressure to give a residue, which was purified by flash column chromatography (80 g silica gel column) eluting with 0 - 5% MeOH in DCM to afford the desired product (280 mg, 39% yield) as a brown solid. LC-MS: m / z 306.2 [M+H] + 。

[0310] Additional intermediates of the present invention were prepared by using the corresponding derivatives. The selected compounds and their corresponding characteristic data are shown in the following table.

Table 12

[0311] Intermediate 68

Chem.

[0312] A mixture of 3,3-difluoropyrrolidine (128.0 mg, 1.2 mmol), 3,3-difluoropyrrolidine (128 mg, 1.2 mmol), and NaBH(OAc)3 (633 mg, 3.0 mmol) in DCM (8 mL) was stirred at 25 °C for 2 h under a N2 atmosphere It was stirred. The reaction mixture was quenched with water (50 mL) and then extracted with EtOAc (2 × 100 mL). The combined organic phases were dried over anhydrous Na2SO4 and then filtered. The filtrate was concentrated. The residue was purified by flash column chromatography (24 g, MeOH 0 - 20% in DCM for 20 minutes) to afford the desired product (156 mg, 73% yield) as a colorless oil. LC-MS: (ESI) m / z 214.2 [M + H] + .

[0313] Additional intermediates of the present invention were prepared by using the corresponding derivatives. The selected compounds and their corresponding property data are shown in the following table.

Table 13

[0314] Intermediate 73

Chem.

[0315] Step 1 To a stirred solution of 5-methyl-2-nitro-pyridine (5.0 g, 36.2 mmol) was added 1-bromopyrrolidine-2,5-dione (6.7 g, 38.0 mmol) and azo-diisobutyronitrile (0.6 g, 3.6 mmol). The reaction mixture was stirred at 80 °C for 4 h under a N2 atmosphere. The mixture was filtered through Celite. The filtrate was concentrated and purified to afford the desired product (4.8 g, 61% yield) as a pale yellow solid. LC-MS: m / z 216.9 [M + H] + .

[0316] Step 2 A stirred mixture of morpholin-3-one (1.4 g, 13.8 mmol) and cesium carbonate (2.2 g, 6.9 mmol) in DMF (15 mL) was stirred at 25 °C for 12 h. To the above mixture was added 5-(bromomethyl)-2-nitro-pyridine (1.0 g, 4.6 m (100 mg, 0.42 mmol) was added at 25 °C. The resulting mixture was stirred at 25 °C for 4 h under a N2 atmosphere. The mixture was poured into water (100 mL) and extracted with dichloromethane (5 × 200 mL). The combined organic phases were dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to give a residue, which was purified to give the desired product (100 mg, 7% yield) as a white solid. LC-MS: m / z 238 [M+H] + .

[0317] Step 3 To a stirred solution of 4-[(6-nitro-3-pyridyl)methyl]morpholin-3-one (40.0 mg, 168 μmol) in ethanol (2 mL), NH4Cl (100 mg, 2.0 mmol) and iron powder (37.6 mg, 674 μmol) were added at 25 °C. The reaction mixture was stirred at 80 °C for 12 h under a N2 atmosphere. The mixture was filtered through a Celite pad. The filtrate was concentrated under reduced pressure to give the desired product (23.0 mg, 65% yield) as a pale yellow solid, which was used directly in the next step without further purification. LC-MS: m / z 208 [M+H] + .

[0318] Intermediate 74 [Chemical Structure Diagram]

[0319] Step 1 A mixture of 6-chloro-2-methyl-pyridine-3-carboxylic acid (2.0 g, 11.6 mmol), piperidin-4-one (1.4 g, 13.9 mmol), N-ethyl-N-isopropyl-propan-2-amine (3.8 g, 29.1 mmol) and HATU (5.3 g, 14.0 mmol) in DCM (25 mL) was stirred at room temperature for 2 h under a N2 atmosphere. The reaction mixture was quenched with water (100 mL) and then extracted with DCM (2×100 mL). The combined organic phases were dried over anhydrous Na2SO4 and then filtered. The filtrate was concentrated. The residue was purified by flash column chromatography (40 g, 0 - 100% EtOAc in PE in 20 min) to afford the desired product (3.5 g, 95% yield) as a yellow oil. LC-MS: m / z 253.1 [M+H] + .

[0320] Step 2 A mixture of 1-(6-chloro-2-methyl-pyridine-3-carbonyl)piperidin-4-one (760 mg, 3.0 mmol), tert-butyl carbamate (421 mg, 3.6 mmol), Pd2(dba)3 (137 mg, 149 μmol), RuPhos (137 mg, 294 μmol) and Cs2CO3 (1.5 g, 4.6 mmol) in dioxane (10 mL) was stirred at 100 °C for 2 h under a N2 atmosphere. The reaction mixture was quenched with water (50 mL) and then extracted with EtOAc (2×100 mL). The combined organic phases were dried over anhydrous Na2SO4 and then filtered. The filtrate was concentrated. The residue was purified by flash column chromatography (12 g, 0 - 100% EtOAc in PE in 10 min) to afford the desired product (430 mg, 36% yield) as a yellow solid. LC-MS: m / z 334.1 [M+H] + .

[0321] Step 3 A mixture of tert-butyl N-[6-methyl-5-(4-oxopiperidine-1-carbonyl)-2-pyridyl]carbamate (1.9 g, 5.9 mmol), cyclopropanamine (681 mg, 11.9 mmol) and NaBH(OAc)3 (3.8 g, 17.9 mmol) in DCM (100 mL) was stirred at 25 °C for 2 h. The reaction mixture was quenched with water (200 mL) and then extracted with DCM (3 × 150 mL). The combined organic phases were dried over anhydrous Na2SO4 and then filtered. The filtrate was concentrated. The residue was purified by flash column chromatography (40 g, 0 - 100% EtOAc in PE over 25 min) to give the desired product (1.3 g, 30% yield) as a yellow solid. LC-MS: m / z 375.1 [M+H] + 。

[0322] Step 4 A mixture of tert-butyl N-[5-[4-(cyclopropylamino)piperidine-1-carbonyl]-6-methyl-2-pyridyl]carbamate (1.3 g, 3.4 mmol) in TFA (6 mL) was stirred at room temperature for 2 h. Anhydrous K2CO3 was added to the reaction mixture and then filtered. The filtrate was concentrated to give the desired product (750 mg, 80% yield) as a yellow solid. LC-MS: m / z 275.1 [M+H] + 。

[0323] Additional intermediates of the present invention were prepared by using the corresponding derivatives.

[0324] The selected compounds and their corresponding property data are shown in the following table.

[0325]

Table 14-1

Table 14-2

[0326] Intermediate 76

Chemical formula

[0327] To a suspension of oxalate of 1,2,3,4,6,7,8,8a-octahydropyrrolo[1,2-a]pyrazine (2.0 g, 15.8 mmol) in DCM / MeCN (5 / 1), anhydrous Na2CO3 (4.6 g, 43.3 mmol) was added. Then the mixture was stirred at room temperature for 16 h. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue and 6-aminopyridine-3-carbaldehyde (1.3 g, 10.6 mmol) were dissolved in DCM (20 mL). Then sodium triacetoxyborohydride (6.0 g, 28.3 mmol) was added to the reaction mixture. The reaction mixture was stirred at room temperature for 16 h. The mixture was quenched with 10 mL of MeOH. Then the mixture was diluted with EtOAc (200 mL). Then the mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (40 g silica gel column, MeOH (5% NH4OH) in DCM 0 - 20%) to give the desired product (720 mg, yield 19%). LC-MS: m / z 233.1 [M+H] + 。

Table 15

[0328] Intermediate 79

Chem.

[0329] Step 1 A mixture of 1-((6-bromopyridin-2-yl)methyl)-4-ethylpiperazine (300 mg, 1.1 mmol), bis(4-methoxybenzyl)amine (327 mg, 1.3 mmol), Pd2(dba)3 (302 mg, 0.3 mmol), RuPhos (306 mg, 0.7 mmol) and Cs2CO3 (718 mg, 2.2 mmol) in dioxane (10 mL) was stirred at 110 °C for 4 h under N2 protection. EtOAc (80 mL) was added to this mixture and it was filtered. The filtrate was concentrated in vacuo to give a residue. The residue was purified by flash column chromatography (4 g silica gel column, MeOH 0 - 10% in DCM) to give the desired product (409 mg, 84% yield) as a pale yellow solid. LC-MS: (ESI) m / z 461.3 [M+H] + .

[0330] Step 2 A mixture of 6-[(4-ethylpiperazin-1-yl)methyl]-N,N-bis[(4-methoxyphenyl)methyl]pyridin-2-amine (130 mg, 282 μmol) in 2,2,2-trifluoroacetic acid (2 mL) was stirred at 50 °C for 5 h. The mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography eluting with methanol containing 0 - 10% dichloromethane to give the desired product (50.0 mg, 80% yield) as a blank solid. LC-MS: (ESI) m / z 221 [M+H] + .

[0331] Intermediate 80 [Chemical formula] To a solution of 1H-pyrazol-3-amine (1.0 g, 12.0 mmol) and tert-butyl 4-(bromomethyl)piperidine-1-carboxylate (3.3 g, 12.0 mmol) in DMF (10 mL), cesium carbonate (11.7 g, 36.1 mmol) was added at 25 °C. The mixture was stirred at 25 °C for 3 h. The organic phase was filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with 0 - 15% methanol in dichloromethane over 20 min to afford the desired product (1.2 g, 35% yield) as a brown solid. LC-MS: m / z 281.2 [M+H] + 。

[0332] By using the corresponding derivatives, additional intermediates of the present invention were prepared. The selected compounds and their corresponding characteristic data are shown in the following table.

[0333]

Table 16

[0334] Synthesis Example 1

Chem.

[0335] CDK4 IC 50 :A;CDK6 IC 50 :B;CDK2 IC 50 :C。

[0336] Synthesis Example 16

Chemical formula

[0337] To a mixture of 1-(6-amino-3-pyridyl)-4-(dimethylamino)piperidin-2-one (30 mg, 128 μmol) and 5-(2-chloropyrimidin-4-yl)-3-isopropyl-pyrazolo[1,5-a]pyridine (38.4 mg, 140.8 μmol) in dioxane (5 mL) were added cesium carbonate (125.1 mg, 384.1 μmol), tris(dibenzylideneacetone)dipalladium(0) (11.7 mg, 12.8 μmol) and RuPhos (11.9 mg, 25.6 μmol). The resulting mixture was stirred at 110 °C for 4 hours under a nitrogen atmosphere. The reaction mixture was extracted with EA (20 mL). The organic phase was washed with water (3 × 20 mL) and brine (3 × 20 mL), and dried over Na2SO4. The mixture was concentrated under reduced pressure and purified by flash column chromatography (DCM / MeOH = 10:1) to obtain the desired product (23.8 mg, yield 39%) as a yellow solid. LC-MS: m / z 471.2 [M+H] + 。

[0338] CDK4 IC 50 :A;CDK6 IC 50 :A;CDK2 IC 50 :B。

[0339] Synthesis Examples 160 and 161

Chemical formula

[0340] N-[5-[4-(Dimethylamino)-1-piperidyl]-2-pyridyl]-4-(3-isopropylpyrazolo[1,5-a]pyridin-5-yl)pyrimidin-2-amine (210 mg, 459.9 μmol) was subjected to chiral separation by SFC (hexane / EtOH / DEA = 60 / 40 / 0.1) using a mobile phase (wavelength: UV 214 nm, column: CHIRALCEL OD-H 5.0 cm ID × 25 cm L, flow rate: 60 mL / min), and Synthesis Example 160 (44.2 mg, yield 21%) was obtained as a yellow solid (LC-MS: m / z 471.2 [M+H] + , ee value > 99%), and Synthesis Example 161 (41.0 mg, yield 19%) was obtained as a yellow solid (LC-MS: m / z 471.2 [M+H] + , ee value 97%).

[0341] Synthesis Example 160, CDK4 IC 50 : A; CDK6 IC 50 : B; CDK2 IC 50 : B.

[0342] Synthesis Example 161, CDK4 IC 50 : A; CDK6 IC 50 : A; CDK2 IC 50 : A.

[0343] Synthesis Example 10

Chemical Structure

[0344] A solution of 1-methylsulfonylpiperidin-4-amine (20.2 mg, 113.5 μmol) and 5-(2-chloro-5-fluoro-pyrimidin-4-yl)-3-isopropyl-pyrazolo[1,5-a]pyridine (30 mg, 103.2 μmol) in anhydrous dioxane (8 mL) was added with tris(dibenzylideneacetone)dipalladium(0) (9.45 mg, 10.3 μmol), dicyclohexyl-[2-(2,6-diisopropoxyphenyl)phenyl]phosphane (9.6 mg, 20.6 μmol) and cesium carbonate (100.8 mg, 309.5 μmol) under a N2 atmosphere. Subsequently, the reaction mixture was stirred at 110 °C for 5 h. The reaction mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography to obtain the desired product (9 mg, yield 20%) as a pale yellow solid. LC-MS: m / z 433.2 [M+H] + .

[0345] CDK4 IC 50 : B; CDK6 IC 50 : C; CDK2 IC 50 : B.

[0346] Synthesis Example 175

Chemical Structure

[0347] A solution of 3-tert-butyl-5-(2-chloropyrimidin-4-yl)pyrazolo[1,5-a]pyridine (40 mg, 139.4 μmol) and 1-(6-amino-3-pyridyl)-4-(dimethylamino)piperidin-2-one (32.6 mg, 139.4 μmol) in anhydrous dioxane (8 mL) was added with tris(dibenzylideneacetone)dipalladium(0) (12.7 mg, 13.95 μmol), RuPhos (13 mg, 27.9 μmol) and cesium carbonate (136.3 mg, 418.4 μmol) under N2 atmosphere. Then, the reaction mixture was stirred at 110 °C for 3 h under N2 atmosphere. The reaction mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography (40 g silica gel column, MeOH containing 0 - 10% DCM) to give the desired product (25 mg, yield 37%) as a yellow solid. LC-MS: m / z 485.3 [M+H] + .

[0348] CDK4 IC 50 : A; CDK6 IC 50 : A; CDK2 IC 50 : A.

Table 17-1

Table 17-2

Table 17-3

Table 17-4

Table 17-5

Table 17-6

Table 17-7

[0349] Synthesis Example 29

Chem.

[0350] Step 1 To a solution of 5-(2-chloro-5-fluoro-pyrimidin-4-yl)-3-isopropyl-pyrazolo[1,5-a]pyridine (150 mg, 515 μmol) and tert-butyl 4-(6-amino-3-pyridyl)-3-oxo-piperazine-1-carboxylate (196 mg, 670 μmol) in dioxane (20 mL) were added tris(dibenzylideneacetone)dipalladium(0) (47.2 mg, 51.6 μmol), RuPhos (24.0 mg) and cesium carbonate (336 mg, 1.0 mmol) at 25 °C. The mixture was then stirred at 110 °C for 2 h. The solvent was removed under reduced pressure. The residue was purified by silica gel chromatography eluting with 0 - 15% MeOH in DCM over 20 min to afford the desired product (240 mg, 85% yield) as a pale yellow solid. LC-MS: m / z 547.2 [M+H] + 。

[0351] Step 2 To a solution of tert-butyl 4-[6-[[5-fluoro-4-(3-isopropylpyrazolo[1,5-a]pyridin-5-yl)pyrimidin-2-yl]amino]-3-pyridyl]-3-oxo-piperazine-1-carboxylate (240 mg, 439 μmol) in DCM (2 mL) was added HCl (2 mL, 2N in EA) at 25 °C. The mixture was stirred for 2 h. The mixture was filtered and the residue was washed with Et2O (10 mL) to afford the desired product (190 mg, 96% yield) as a yellow solid. LC-MS: m / z 447.2 [M+H] + 。

[0352] CDK4 IC 50 : A; CDK6 IC 50 : B; CDK2 IC 50: B.

[0353] Synthesis Example 204 [Chemical formula]

[0354] Step 1 To a solution of tert-butyl N-[1-(6-amino-3-pyridyl)-2-oxo-4-piperidyl]-N-methyl-carbamate (399.4 mg, 1.2 mmol) in dioxane (30 mL) were added 5-(2-chloropyrimidin-4-yl)-3-isopropyl-pyrazolo[1,5-a]pyridine (340 mg, 1.2 mmol), tris(dibenzylideneacetone)dipalladium(0) (114.1 mg, 124.6 μmol), RuPhos (116.3 mg, 249.3 μmol) and cesium carbonate (812.3 mg, 2.4 mmol). The reaction mixture was stirred at 110 °C for 16 h in a 15 mL sealed tube. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (12 g silica gel, MeOH 0 - 10% in DCM) to give the desired product (544 mg, 78% yield) as a yellow oil. LC-MS: m / z 557.3 [M+H] + .

[0355] Step 2 To a solution of tert-butyl N-[1-[6-[[4-(3-isopropylpyrazolo[1,5-a]pyridin-5-yl)pyrimidin-2-yl]amino]-3-pyridyl]-2-oxo-4-piperidyl]-N-methyl-carbamate (544 mg, 977.2 μmol) in DCM (10 mL) was added hydrogen chloride solution (2.0 M in ethyl acetate) (5 mL) at 25 °C. The mixture was stirred at 25 °C for 2 h. The mixture was filtered and concentrated in vacuo to give the desired product (430 mg, 96% yield) as a yellow solid. LC-MS: m / z 457.3 [M+H] + .

[0356] CDK4 IC 50 : A; CDK6 IC 50: A; CDK2 IC 50 : A。

[0357] Synthesis Examples 205 and 206

Chemical Formula

[0358] 1-[6-[[4-(3-Isopropylpyrazolo[1,5-a]pyridin-5-yl)pyrimidin-2-yl]amino]-3-pyridyl]-4-(methylamino)piperidin-2-one (380 mg, 832 μmol) was subjected to chiral separation using an ID column with a mobile phase (MeOH / ACN / DEA = 50 / 50 / 0.1) (wavelength: UV 214 nm, column: CHIRALPAK ID-H 25 cm L 5.0 cm ID 10 μm, flow rate: 60 g / min)), and Synthesis Example 205 (132 mg, yield 34%) was obtained as a white solid (LC-MS: m / z 457.3 [M+H] + , ee value: >98%), and Synthesis Example 206 (120 mg, yield 31%) was obtained as a white solid (LC-MS: m / z 457.3 [M+H] + , ee value: >98%).

[0359] Synthesis Example 205, CDK4 IC 50 : A; CDK6 IC 50 : A; CDK2 IC 50 : A。

[0360] Synthesis Example 206, CDK4 IC 50 : A; CDK2 IC 50 : A。

Table 18-1

Table 18-2

Table 18-3

Table 18-4

Table 18-5

Table 18-6

Table 18-7

[0361] Synthesis Example 39

Chem.

[0362] Step 1 To a solution of 5-iodopyridin-2-amine (100 mg, 454 μmol) and tert-butyl N-(5-oxopyrrolidin-3-yl)carbamate (91.1 mg, 454 μmol) in dioxane (5 mL), CuI (8.6 mg, 45.4 μmol), tripotassium phosphate (289 mg, 1.3 mmol) and (1R,2R)-N1,N2-dimethylcyclohexane-1,2-diamine (12.9 mg, 90.9 μmol) were added under N2 atmosphere. The reaction mixture was stirred at 110 °C for 8 h. The mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (eluting with 0 - 35% MeOH in DCM) to give the desired product (90.0 mg, 67% yield) as a yellow solid. LC-MS: m / z 292.2 [M+H] + 。

[0363] Step 2 To a solution of tert-butyl N-[1-(6-amino-3-pyridyl)-5-oxo-pyrrolidin-3-yl]carbamate (90.0 mg, 307 μmol) and 5-(2-chloropyrimidin-4-yl)-3-isopropyl-pyrazolo[1,5-a]pyridine (83.9 mg, 307 μmol) in dioxane (2 mL) were added cesium carbonate (300 mg, 923 μmol), dicyclohexyl-[2-(2,6-diisopropoxyphenyl)phenyl]phosphane (28.7 mg, 61.5 μmol) and tris(dibenzylideneacetone)dipalladium(0) (28.1 mg, 30.7 μmol) under N2 atmosphere. The reaction mixture was stirred at 110 °C for 8 h. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography eluting with 0 - 10% MeOH in DCM to give the desired product (90.0 mg, 55% yield) as a yellow solid. LC-MS: m / z 529.3 [M+H] + .

[0364] Step 3 To a solution of tert-butyl N-[1-[6-[[4-(3-isopropylpyrazolo[1,5-a]pyridin-5-yl)pyrimidin-2-yl]amino]-3-pyridyl]-5-oxo-pyrrolidin-3-yl]carbamate (90.0 mg, 170 μmol) in DCM (10 mL) was added HCl (4N, 0.1 mL) in EA. The reaction mixture was stirred at 25 °C for 2 h. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography eluting with 0 - 10% MeOH in DCM to give the desired product (70.0 mg, 95% yield) as a yellow solid. LC-MS: m / z 429.2 [M+H] + .

[0365] Step 4 4-Amino-1-[6-[[4-(3-isopropylpyrazolo[1,5-a]pyridin-5-yl)pyrimidin-2-yl]amino]-3-pyridyl]pyrrolidin-2-one (50.0 mg, 116 μmol) and formaldehyde (14.0 mg, 466 μmol) in DCM (5 mL) were stirred at 25 °C for 0.5 h. Then, NaBH(OAc)3 (74.1 mg, 350 μmol) was added to the above solution. The mixture was stirred at 25 °C for 8 h. The reaction was concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product (15.9 mg, 29% yield) as a yellow solid. LC-MS: m / z 457.2 [M+H] + 。

[0366] CDK4 IC 50 : A; CDK6 IC 50 : B; CDK2 IC 50 : B。

[0367] Synthesis Example 40

Chemical formula

[0368] Step 1 To a solution of 1-[6-[[4-(3-isopropylpyrazolo[1,5-a]pyridin-5-yl)pyrimidin-2-yl]amino]-3-pyridyl]piperazin-2-one (80 mg, 186 μmol) and tert-butyl 3-oxoazetidine-1-carboxylate (63.9 mg, 373 μmol) in DCM (10 mL), sodium triacetoxyborohydride (118 mg, 560 μmol) and acetic acid (16.8 mg, 280 μmol) were added. The reaction mixture was stirred at 15 °C for 12 h. The reaction mixture was washed with H2O (5 ml) and brine (5 mL). The organic layer was concentrated and the residue was purified by flash column chromatography (12 g silica gel column, 0-10% MeOH in DCM) to give the desired product (72.0 mg, 66% yield) as a yellow solid. LC-MS: m / z 584.3 [M+H] + 。

[0369] Step 2 A solution of tert-butyl 3-[4-[6-[[4-(3-isopropylpyrazolo[1,5-a]pyridin-5-yl)pyrimidin-2-yl]amino]-3-pyridyl]-3-oxo-piperazin-1-yl]azetidine-1-carboxylate (70.0 mg, 119 μmol) in HCl / 1,4-dioxane (1N, 10 mL) was stirred at 25 °C for 2 hours. The mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography (20 g silica gel column, 0 - 10% MeOH in DCM) to give the desired product (12.5 mg, 21% yield) as a yellow solid. LC-MS: m / z 484.2 [M+H] + .

[0370] CDK4 IC 50 : A; CDK6 IC 50 : B; CDK2 IC 50 : B.

Table 19

[0371] Synthesis Example 43

Chemical formula

[0372] A solution of 4-[6-[[5-fluoro-4-(3-isopropylpyrazolo[1,5-a]pyridin-5-yl)pyrimidin-2-yl]amino]-3-pyridyl]-1,4-diazepan-5-one (40.0 mg, 86.8 μmol), oxetan-3-one (9.0 mg, 124 μmol) and NaBH3CN (20.0 mg, 300 μmol) in methanol (8 mL) was stirred at 25 °C for 16 hours. The mixture was filtered and the filtrate was concentrated. The residue was purified by preparative HPLC to give the desired product (12.3 mg, 27% yield) as a yellow solid. LC-MS: m / z 517.3 [M+H] + .

[0373] CDK4 IC50 : A; CDK6 IC 50 : B; CDK2 IC 50 : B。

Table 20

[0374] Synthesis Example 49

Chem.

[0375] 4-[6-[[5-Fluoro-4-(3-isopropylpyrazolo[1,5-a]pyridin-5-yl)pyrimidin-2-yl]amino]-3-pyr ridyl]-1,4-diazepan-5-one (20 mg, 43.4 μmol), bromocyclopropane (4.7 mg, 39.0 μmol) and silver carbonate (7.2 mg, 43.4 μmol) in methylbenzene (5 mL) was stirred at 70 °C for 5 h. The mixture was diluted with water (15 mL) and extracted with DCM (2 × 20 mL). The combined organic phases were concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product (2 mg, 9% yield) as a yellow solid. LC-MS: m / z 501.2 [M+H] + 。

[0376] CDK4 IC 50 : A; CDK2 IC 50 : C。

[0377] Synthesis Example 50

Chem.

[0378] To a stirred solution of 1-[6-[[5-fluoro-4-(3-isopropylpyrazolo[1,5-a]pyridin-5-yl)pyrimidin-2-yl]amino]-3-pyridyl]-1,4-diazepan-2-one (22.0 mg, 47.7 μmol) and TEA (10 mg, 100 μmol) in DCM (5 mL) was added methylsulfonyl chloride (7.3 mg, 64.2 μmol). The reaction mixture was stirred at 20 °C for 16 h. The reaction mixture was quenched with water (3 mL) and then extracted with ethyl acetate (2 × 5 mL). The combined organic phases were dried over sodium sulfate and then filtered. The filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC to afford the desired product (5.0 mg, 17% yield) as a yellow solid. LC-MS: m / z 539.2 [M+H] + 。

[0379] CDK4 IC 50 :A;CDK6 IC 50 :B;CDK2 IC 50 :B。

Table 21

[0380] Synthesis Example 52

Chemical Structure

[0381] A solution of 5-[6-[[5-fluoro-4-(3-isopropylpyrazolo[1,5-a]pyridin-5-yl)pyrimidin-2-yl]amino]-3-pyridyl]azepan-4-one (30.0 mg, 65.2 μmol) and potassium carbonate (27.0 mg, 195 μmol) in acetonitrile (5 mL) was treated with 2-iodoethanol (13.4 mg, 78.3 μmol) at 25 °C. The reaction mixture was stirred at 50 °C for 16 h. The mixture was quenched with water (20 mL) and then extracted with ethyl acetate (2 × 10 mL). The combined organic phases were dried over sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC to afford the desired product (5.0 mg, 15% yield) as a yellow solid. LC-MS: m / z 505.3 [M+H] + 。

[0382] CDK4 IC 50 :A;CDK6 IC 50 :B;CDK2 IC 50 :B。

Table 22

[0383] Synthesis Example 55

Chemical formula

[0384] 4-(3-Isopropylpyrazolo[1,5-a]pyridin-5-yl) in DCM (4 mL) A solution of 1-(4-Piperidylmethyl)-1H-pyrazol-3-yl)-N-(pyrimidin-2-yl)amine (20.0 mg, 48.0 μmol), glycolic acid (5.4 mg, 72.0 μmol), 4-methylmorpholine (4.8 mg, 48.0 μmol), EDCI (13.8 mg, 71.9 μmol) and HOBT (9.7 mg, 72.0 μmol) was stirred at 20 °C for 5 h. The mixture was diluted with water (15 mL) and extracted with DCM (2 × 20 mL). The combined organic phases were concentrated under reduced pressure. The residue was purified to give the desired product (3.7 mg, 16% yield) as a yellow solid. LC-MS: (ESI) m / z 475.3 [M+H] + .

[0385] CDK4 IC 50 : B; CDK2 IC 50 : D.

Table 23

[0386] Synthesis Example 57

Chemical formula

[0387] A mixture of 5-(2-chloropyrimidin-4-yl)-3-isopropyl-6,7-dihydro-4H-imidazo[4,5-c]pyridine (66.0 mg, 237 μmol), 4-(6-amino-3-pyridyl)tetrahydropyran-3-one (54.8 mg, 285 μmol), tris(dibenzylideneacetone)dipalladium(0) (21.7 mg, 23.7 μmol), dicyclohexyl-[2-(2,6-diisopropoxyphenyl)phenyl]phosphane (22.1 mg, 47.5 μmol) and cesium carbonate (232 mg, 712 μmol) in dioxane (6.5 mL) was stirred at 110 °C for 6 h under a N2 atmosphere. The reaction mixture was quenched with water (50 mL) and then extracted with EtOAc (2 × 100 mL). The combined organic phases were dried over anhydrous Na2SO4 and then filtered. The filtrate was concentrated. The residue was purified by preparative HPLC to give the desired composition (9.9 mg, yield 19%) as a white solid. LC-MS: (ESI) m / z 435.3 [M+H] + .

[0388] CDK4 IC 50 : A; CDK6 IC 50 : C; CDK2 IC 50 : B.

Table 24-1

Table 24-2

Table 24-3

[0389] Synthesis Example 58

Chemical Structure

[0390] A mixture of 5-(6-chloro-3-fluoro-2-pyridyl)-3-isopropyl-2-methyl-pyrazolo[3,4-c]pyridine (30.0 mg, 98.4 μmol), 5-[(4-ethylpiperazin-1-yl)methyl]pyridin-2-amine (26.0 mg, 118 μmol), Pd2(dba)3 (4.5 mg, 4.9 μmol), RuPhos (4.5 mg, 9.8 μmol) and Cs2CO3 (64.1 mg, 196 μmol) in dioxane (4 mL) was stirred at 100 °C for 1 h under a N2 atmosphere. The reaction mixture was quenched with water (50 mL) and extracted with EA (2 × 100 mL). The combined organic phases were dried over anhydrous Na2SO4 and then filtered. The filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product (3.0 mg, 6% yield) as a yellow solid. LC-MS: (ESI) m / z 490.1 [M+H] + 。

[0391] CDK4 IC 50 : C; CDK6 IC 50 : D。

[0392] Synthesis Example 59

Chemical Structure

[0393] Step 1 A mixture of 2-chloro-4-(7-fluoro-3,3-dimethyl-2,3-dihydrobenzofuran-5-yl)pyrimidine (50.0 mg, 180 μmol), tert-butyl 4-(6-aminopyridin-3-yl)-3-oxopiperazine-1-carboxylate (26.0 mg, 216 μmol), Pd2(dba)3 (10.0 mg, 10.0 μmol), RuPhos (10.0 mg, 19.0 μmol) and Cs2CO3 (130 mg, 400 μmol) in dioxane (8 mL) was stirred at 100 °C for 1 h under a N2 atmosphere. The reaction mixture was quenched with water (50 mL) and extracted with EA (2 × 100 mL). The combined organic phases were dried over anhydrous Na2SO4 and then filtered. The filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product (60 mg, 50%). LC-MS: (ESI) m / z 535.2 [M+H] + 。

[0394] Step 2 A solution of tert-butyl 4-(6-((4-(7-fluoro-3,3-dimethyl-2,3-dihydrobenzofuran-5-yl)pyrimidin-2-yl)amino)pyridin-3-yl)-3-oxopiperazine-1-carboxylate (60.0 mg, 112 μmol) in HCl / 1,4-dioxane (1 N, 10 mL) was stirred at 25 °C for 2 h. The mixture was concentrated under reduced pressure and the residue was purified by flash column chromatography (20 g silica gel column, MeOH 0 - 10% in DCM) to give the desired product (38.0 mg, 78% yield) as a yellow solid. LC-MS: m / z 435.2 [M+H] + 。

[0395] CDK4 IC 50 : A; CDK6 IC 50 : B; CDK2 IC 50 : B。

Table 25-1

Table 25-2

[0396] Synthesis Example 60 [Chemical formula]

[0397] Step 1 To a solution of 5-bromo-3-methoxypyrazine-2-amine (500 mg, 2.5 mmol) in EtOH (10 mL), 2-chloro-3-methylbutanal (443 mg, 3.68 mmol) was added dropwise at 25 °C. The mixture was stirred at 80 °C for 2 hours. The mixture was concentrated to obtain a residue. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1~5:1) to obtain the desired product (25.0 mg, yield 4%) as a yellow solid. LC-MS: (ESI) m / z 270.0 [M+H] + .

[0398] Step 2 To a solution of 6-bromo-3-isopropyl-8-methoxyimidazo[1,2-a]pyrazine (25.0 mg, 0.09 mmol) in dioxane (5 mL), B2Pin2 (150 mg, 0.6 mmol), KOAc (27.0 mg, 0.3 mmol) and Pd(DPPF)Cl2 (15.0 mg, 0.02 mmol) were added at 25 °C under a N2 atmosphere. The mixture was stirred at 110 °C for 3 hours. The mixture was filtered through diatomaceous earth and concentrated to obtain the desired product (30 mg) as a yellow solid, which was used in the next step without purification. LC-MS: (ESI) m / z 236.1 [M+H] + .

[0399] Step 3 A solution of (3-isopropyl-8-methoxyimidazo[1,2-a]pyrazin-6-yl)boronic acid (25.0 mg, 0.1 mmol) and 4-chloro-N-(5-((4-ethylpiperazin-1-yl)methyl)pyridin-2-yl)-5-fluoropyrimidin-2-amine (37.0 mg, 0.2 mmol) in dioxane (4 mL) and water (1 mL) was added with K2CO3 (30.0 mg, 0.2 mmol) and Pd(dppf)Cl2 (15.0 mg, 0.02 mmol) at 25 °C under a N2 atmosphere. The mixture was stirred at 100 °C for 3 h. The mixture was filtered through celite and concentrated. The residue was purified by preparative HPLC to give the desired product (5.1 mg, yield 9%) as a yellow solid. LC-MS: (ESI) m / z 506.3 [M+H] + 。

[0400] CDK4 IC 50 :B;CDK6 IC 50 :C。

[0401] Synthesis Example 61

Chemical formula

[0402] To a mixture of 6-(2-chloropyrimidin-4-yl)-4-fluoro-1-isopropyl-imidazo[4,5-c]pyridine (22.0 mg, 75.4 μmol) in 1,4-dioxane (5 mL), 4-(6-amino-3-pyridyl)morpholin-3-one (16.0 mg, 82.8 μmol), dicyclohexyl-[2-(2,6-diisopropoxyphenyl)phenyl]phosphane (12.0 mg, 25.7 μmol), tris(dibenzylideneacetone)dipalladium (11.0 mg, 12.0 μmol) and cesium carbonate (80.0 mg, 245 μmol) were added and the mixture was degassed with N2. The resulting mixture was stirred at 100 °C for 12 h. The mixture was concentrated and purified by column (DCM / MeOH = 20 / 1 - 8 / 1) to give the desired product (1 mg, yield 3%). LC-MS: m / z 449.1 [M+H] + 。

[0403] CDK4 IC 50 :A;CDK6 IC 50 :B;CDK2 IC 50 :B。

[0404] Synthesis Example 287

Chem.

[0405] To a solution of 5-(4-isopropylpiperazin-1-yl)pyridin-2-amine (124.6 mg, 565 μmol) and 6-(2-chloropyrimidin-4-yl)-4-fluoro-1-isopropyl-imidazo[4,5-c]pyridine (150 mg, 514 μmol) in dioxane (15 mL) were added Pd2(dba)3 (47.1 mg, 51 μmol), RuPhos (47.9 mg, 102 μmol) and Cs2CO3 (502.6 mg, 1.5 mmol). The mixture was stirred at 110 °C for 3 h under N2. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography eluting with 0 - 10% MeOH in DCM to give the desired product (107.2 mg, 43% yield) as a yellow solid. LC-MS: m / z 476.2 [M+H] + 。

[0406] CDK4 IC 50 :A;CDK6 IC 50 :B;CDK2 IC 50 :C。

[0407] Synthesis Example 301

Chem.

[0408] Step 1 A solution of 6-(2-chloropyrimidin-4-yl)-4-fluoro-1-isopropyl-2-methyl-imidazo[4,5-c]pyridine (150 mg, 490.6 μmol) and tert-butyl N-[1-(6-amino-3-pyridyl)-2-oxo-3-piperidyl]carbamate (150.3 mg, 490.6 μmol) in dioxane (5 mL) was added with cesium carbonate (479.5 mg, 1.4 mmol), RuPhos (45.7 mg, 98.1 μmol) and tris(dibenzylideneacetone)dipalladium(0) (44.9 mg, 49.0 μmol). The reaction mixture was stirred at 110 °C for 8 h. The reaction mixture was purified by column chromatography eluting with 0 - 4% MeOH in DCM in 15 min to give the desired product (170 mg, 60% yield) as a yellow solid.

[0409] Step 2 HCl (2 M, 738 μL) was added to a solution of tert-butyl N-[1-[6-[[4-(4-fluoro-1-isopropyl-2-methyl-imidazo[4,5-c]pyridin-6-yl)pyrimidin-2-yl]amino]-3-pyridyl]-2-oxo-3-piperidyl]carbamate (170 mg, 295.3 μmol) in DCM (5 mL). The reaction mixture was stirred at 25 °C for 3 h. The reaction mixture was concentrated under reduced pressure to give the desired product (140 mg, 99% yield) as a yellow solid.

[0410] Step 3 Formaldehyde (50.5 mg, 1.6 mmol) was added to a solution of 3-amino-1-[6-[[4-(4-fluoro-1-isopropyl-2-methyl-imidazo[4,5-c]pyridin-6-yl)pyrimidin-2-yl]amino]-3-pyridyl]piperidin-2-one (80 mg, 168.2 μmol) in THF (5 mL). After 1 h, sodium cyanoborohydride (31.7 mg, 504.7 μmol) was added to the above solution. The reaction mixture was stirred at 25 °C for 2 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC to give the desired product (5.7 mg, 6% yield) as a yellow solid. LC-MS: m / z 504.3 [M + H]+ .

[0411] CDK4 IC 50 :A;CDK6 IC 50 :A;CDK2 IC 50 :D.

[0412] Synthesis Examples 302 and 303 [Chemical Formula]

[0413] 3-(Dimethylamino)-1-[6-[[4-(4-Fluoro-1-isopropyl-2-methyl-imidazo[4,5-c]pyridin-6-yl)pyrimidin-2-yl]amino]-3-pyridyl]piperidin-2-one (170 mg, 337.6 μmol) was separated by SFC (column: Daicel CHIRALPAK OD_3, 3 × 150 mm, 3 μm; mobile phase: A / B: CO2 / MeOH(0.1%DEA) = 70 / 30; flow rate: 2.0 mL / min) to obtain Synthesis Example 302 (50 mg, yield 29%) (LC-MS: m / z 504.3 [M+H] + , ee value: 94%) and Synthesis Example 303 (50 mg, yield 29%) (LC-MS: m / z 504.3 [M+H] + , ee value: 94%) as white solids.

[0414] Synthesis Example 302, CDK4 IC 50 :A;CDK2 IC 50 :D.

[0415] Synthesis Example 303, CDK4 IC 50 :A;CDK2 IC 50 :D. [Table 26-1] [Table 26-2] [Table 26-3]

Table 26-4

Table 26-5

Table 26-6

[0416] Synthesis Example 62

Chem.

[0417] To a mixture of 1-(6-amino-3-pyridyl)-4-(dimethylamino)piperidin-2-one (85.0 mg, 363 μmol), 6-(2-chloro-5-fluoro-pyrimidin-4-yl)-1-isopropyl-4-methoxy-2-methyl-imidazo[4,5-c]pyridine (122 mg, 363 μmol) and 2-dicyclohexylphosphino-2′,6′-di-isopropoxy-1,1′-biphenyl (33.0 mg, 72.6 μmol) in dioxane (10 mL) were added Cs2CO3 (355 mg, 1.1 mmol) and tris(dibenzylideneacetone)dipalladium (33.0 mg, 36.3 μmol), and the resulting mixture was stirred at 110 °C for 4 h under a nitrogen atmosphere. The reaction mixture was filtered. The filtrate was concentrated in vacuo to give a residue. The residue was purified by a reverse-phase column (C18, 20 g) eluting with (ACN: water (0.1% formic acid) = 1:10) to give the desired product (9.8 mg, 5% yield) as a white solid. LC-MS: m / z 534.3 [M+H] + 。

[0418] CDK4 IC 50 : A; CDK6 IC 50 : B; CDK2 IC 50 : D。

[0419] Synthesis Examples 62-1 and 62-2

Chemical Structure

[0420] 4-(Dimethylamino)-1-[6-[[5-fluoro-4-(1-isopropyl-4-methoxy-2-methyl-imidazo[4,5-c]pyridin-6-yl)pyrimidin-2-yl]amino]-3-pyridyl]piperidin-2-one (7.9 mg, 14.8 μmol) was purified by chiral-HPLC (apparatus: SFC 80, column: Daicel CHIRALPAK OJ-H 250 mm 20 mm ID, 5 μm, mobile phase: CO2 / MeOH (0.2% NH4OH) = 50 / 50, flow rate: 45 g / min, wavelength: UV214 nm, temperature: 35 °C) to obtain isomer 2: (2.3 mg, yield 29%) (LC-MS: m / z 534.3 [M+H] + , RT = 12.72 min, ee value > 99%), isomer 1: (1.6 mg, yield 20%) (LC-MS: m / z 534.3 [M+H] + , RT = 10.95 min, ee value > 99%).

[0421] Example 62-1, CDK4 IC 50 : A; CDK6 IC 50 : B; CDK2 IC 50 : D.

[0422] Example 62-2, CDK4 IC 50 : A; CDK6 IC 50 : B; CDK2 IC 50 : D.

Table 27-1

Table 27-2

Table 27-3

Table 27-4

Table 27-5

Table 27-6

Table 27-7

[0423] Synthesis Example 106

Chem.

[0424] Step 1 To a solution of 6-(2-chloro-5-fluoro-pyrimidin-4-yl)-1-isopropyl-4-methoxy-2-methyl-imidazo[4,5-c]pyridine (70.0 mg, 208 μmol) in dry dioxane (10 mL) were added tert-butyl 4-(6-amino-3-pyridyl)piperazine-1-carboxylate (69.6 mg, 250 μmol), tris(dibenzylideneacetone)dipalladium(0) (19.0 mg, 20.8 μmol), (5-diphenylphosphanyl-9,9-dimethyl-xanthen-4-yl)-diphenylphosphane (24.1 mg, 41.7 μmol) and cesium carbonate (203 mg, 625 μmol). The mixture was stirred at 110 °C for 16 h. The reaction mixture was filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography (4 g of silica gel, MeOH 0 - 15% in DCM in 20 min) to give the desired product (92.0 mg, yield: 76%) as a white solid. LC-MS: m / z 578.2 [M+H] + 。

[0425] Step 2 A solution of tert-butyl 4-[6-[[5-fluoro-4-(1-isopropyl-4-methoxy-2-methyl-imidazo[4,5-c]pyridin-6-yl)pyrimidin-2-yl]amino]-3-pyridyl]piperazine-1-carboxylate (92.0 mg, 159 μmol) in dry DCM (5 mL) was treated with hydrogen chloride (4 N in 1,4-dioxane) (5 mL). The reaction mixture was stirred at 25 °C for 2 h. The mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC to afford the desired product (10.8 mg, 14% yield) as a yellow solid. LC-MS: m / z 478.2 [M+H] + 。

[0426] CDK4 IC 50 :A;CDK6 IC 50 :B;CDK2 IC 50 :D。

Table 28-1

Table 28-2

[0427] Synthesis Example 121

Chemical Structure

[0428] A solution of 1-[6-[[5-fluoro-4-(1-isopropyl-4-methoxy-2-methyl-imidazo[4,5-c]pyridin-6-yl)pyrimidin-2-yl]amino]-3-pyridyl]piperazin-2-one (50.0 mg, 101 μmol) and 1-methylsulfonylethylene (21.6 mg, 203 μmol) in DMF (5 mL) was added with DIPEA (30.9 mg, 305 μmol). The reaction mixture was stirred at 25 °C for 48 h. Then the reaction mixture was diluted with H2O (30 mL) and extracted with EA (3×10 mL). The organic layer was concentrated under reduced pressure. The residue was purified by preparative HPLC eluting with acetonitrile in water (0.1% FA) 22% - 24% in 6.0 min to give the desired product (2.7 mg, 4% yield) as a yellow solid. LC-MS: m / z 598.3 [M+H] + 。

[0429] CDK4 IC 50 : A; CDK6 IC 50 : B; CDK2 IC 50 : D。

Table 29

[0430] Synthesis Example 127

Chemical Structure

[0431] To a solution of 1-[6-[[5-fluoro-4-(1-isopropyl-4-methoxy-2-methyl-imidazo[4,5-c]pyridin-6-yl)pyrimidin-2-yl]amino]-3-pyridyl]piperazin-2-one (0.2 g, 305 μmol) and TEA (92.6 mg, 915 μmol) in DCM (5 mL) was added dropwise methylsulfonyl chloride (52.4 mg, 457 μmol) at 0 °C. The mixture was stirred at 0 °C for 0.5 h. The mixture was concentrated and purified by preparative HPLC to give the desired product (7.8 mg, 4% yield) as a yellow solid. LC-MS: m / z 570.2 [M+H] + 。

[0432] CDK4 IC 50 :A;CDK6 IC 50 :B;CDK2 IC 50 :C。

[0433] Synthesis Example 128

Chem.

[0434] To a solution of 1-[6-[[5-fluoro-4-(1-isopropyl-4-methoxy-2-methyl-imidazo[4,5-c]pyridin-6-yl)pyrimidin-2-yl]amino]-3-pyridyl]piperazin-2-one (100 mg, 203 μmol) and acetaldehyde (17.9 mg, 406 μmol) in DCM (5 mL) was added NaB(OAc)3H (129 mg, 610 μmol). The mixture was stirred at 25 °C for 1 h. The mixture was diluted with water (10 mL) and extracted with DCM (2 × 10 mL). The combined organic layers were dried over Na2SO4, filtered, concentrated, and purified by preparative HPLC to give the desired product (16.0 mg, yield 15%) as a yellow solid.

[0435] CDK4 IC 50 :A;CDK6 IC 50 :B;CDK2 IC 50 :D。

Table 30-1

Table 30-2

[0436] Synthesis Example 138

Chem.

[0437] Step 1 A stirred solution of tert-butyl 3-oxoazetidine-1-carboxylate (26.1 mg, 152 μmol) in methanol (5 mL) was added to 1-[6-[[5-fluoro-4-(1-isopropyl-4-)methoxy-2-methyl-imidazo[4,5-c]pyridin-6-yl)pyrimidine-2-yl]amino]-3-pyridyl]piperazin-2-one ( 50.0 mg, 101 μmol). The reaction mixture was stirred at 25 °C for 1 h under a N2 atmosphere. Sodium cyanoborohydride (12.8 mg, 203 μmol) was added to the above mixture. The reaction mixture was stirred at 25 °C for 11 h under a N2 atmosphere. The mixture was concentrated and purified by flash column chromatography (4 g silica gel column), eluting with DCM / MeOH containing 0 - 6% MeOH to give the desired product (25.0 mg, 38% yield) as a yellow solid. LC-MS: m / z 647.3 [M+H] + .

[0438] Step 2 HCl (1 N in dioxane, 3 mL) was added to a stirred solution of tert-butyl 3-[4-[6-[[5-fluoro-4-(1-isopropyl-4-methoxy-2-methyl-imidazo]4,5-c]pyridin-6-yl)pyrimidine-2-yl]amino]-3-pyridyl]-3-oxo-piperazin-1-yl]azetidine-1-carboxylate (20.0 mg, 30.9 μmol) in DCM (3 mL). The reaction mixture was stirred at 25 °C for 1 h under a N2 atmosphere. The mixture was concentrated and purified by preparative HPLC to give the desired product (1.8 mg, 11% yield) as a yellow solid. LC-MS: m / z 547.2 [M+H] + .

[0439] CDK4 IC 50 : A; CDK6 IC 50 : B; CDK2 IC 50 : C.

Table 31

[0440] Synthesis Example 140 [Chemical formula]

[0441] To a mixture of 2-(dimethylamino)acetic acid (6.9 mg, 67.2 μmol) in DCM (6 mL), DIPEA (82.3 mg, 636 μmol) was added. The mixture was stirred at 20 °C for 10 minutes. To the above mixture, N-[4-(1-isopropyl-4-methoxy-imidazo[4,5-c]pyridin-6-yl)pyrimidin-2-yl]-5,6,7,8-tetrahydro-1,6-naphthyridin-2-amine (28.0 mg, 67.2 μmol) and HATU (25.7 mg, 67.3 μmol) were added. The mixture was stirred at 20 °C for 12 hours. The mixture was quenched with water (1 mL) and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain the desired product (14.5 mg, 43% yield) as a yellow solid. LC-MS: (ESI) m / z 502.2 [M+H] + .

[0442] CDK4 IC 50 : A; CDK6 IC 50 : B; CDK2 IC 50 : D. [Table 32]

[0443] Synthesis Example 143 [Chemical formula]

[0444] Step 1 A solution of 2-chloro-3-methyl-butan-al (1.1 g, 9.1 mmol) and 5-bromopyrazin-2-amine (1.6 g, 9.1 mmol) in ethylene glycol (10 mL) was stirred at 120 °C for 16 h. The mixture was diluted with water (25 mL) and extracted with EtOAc (2 × 25 mL). The combined organic phases were concentrated under reduced pressure. The residue was purified by FCC (20 g silica gel, 0 - 50% EtOAc in PE) to give the desired product (200 mg, 9% yield) as a yellow solid. LC-MS: (ESI) m / z 240.2 [M+H] + .

[0445] Step 2 A solution of 6-bromo-3-isopropyl-imidazo[1,2-a]pyrazine (480 mg, 999 μmol), (2-aminopyrimidin-4-yl)boronic acid (138 mg, 999.5 μmol), K2CO3 (276 mg, 2.0 mmol) and Pd(DPPF)Cl2 (73.1 mg, 99.9 μmol) in dioxane (10 mL) and H2O (0.5 mL) was stirred at 110 °C for 16 h under N2. The mixture was concentrated under reduced pressure. The res idue was purified by FCC (20 g silica gel, 0 - 10% MeOH in DCM) to give the desired product (77.0 mg, 30% yield) as a dark solid. LC-MS: (ESI) m / z 255.1 [M+H] + .

[0446] Step 3 A solution of 4-(3-isopropylimidazo[1,2-a]pyrazin-6-yl)pyrimidine-2-amine (77.0 mg, 302 μmol), tert-butyl 4-(6-chloro-3-pyridyl)-3-oxo-piperazine-1-carboxylate (94.4 mg, 302 μmol), Cs2CO3 (197 mg, 605 μmol), RuPhos (28.2 mg, 60.5 μmol) and Pd2(dba)3 (27.7 mg, 30.2 μmol) in dioxane (8 mL) was stirred at 110 °C for 16 h under N2. The mixture was concentrated under reduced pressure. The residue was purified by FCC (12 g silica gel, MeOH 0 - 10% in DCM) to give the desired product (95.0 mg, 59% yield) as a yellow solid. LC-MS: (ESI) m / z 530.2 [M+H] + 。

[0447] Step 4 HCl (4N, 180 μL) was added to a solution of tert-butyl 4-[6-[[4-(3-isopropylimidazo[1,2-a]pyrazin-6-yl)pyrimidine-2-yl]amino]-3-pyridyl]-3-oxo-piperazine-1-carboxylate (95.0 mg, 179 μmol) in DCM (8 mL) at 25 °C. The reaction mixture was stirred at 25 °C for 4 h. The mixture was concentrated under reduced pressure. The residue was purified by preparative reverse-phase HPLC to give the desired product (19.4 mg, 25% yield) as a yellow solid. LC-MS: (ESI) m / z 430.2 [M+H] + 。

[0448] CDK4 IC 50 :B;CDK2 IC 50 :D。

[0449] Comparative Synthesis Example 1 Compound 10 disclosed in CN109503573A was prepared. As shown in the following table, this compound showed very weak activity in the CDK4 and CDK6 assays.

Table 33

Table 34-1

Table 34-2

Table 34-3

Claims

1. Compounds represented by structural formula (I) 【Chemistry 1】 (Wherein, ring A is 【Chemistry 2】 and Ring B is a bond, a 3- to 10-membered heterocyclyl, or a 5- to 10-membered heteroaryl; Ring C is a 5- to 6-membered heteroaryl, a 5- to 10-membered heterocyclyl, a phenyl, are 5-10 membered bridged bicyclic groups, each of which is １２ and optionally substituted with The linker L is a bond, -(CH ２ ) ｑ -, -(CH ２ ) ｑ O-, -NR ａ (CH ２ ) ｑ -, -C(O)-, -C(O)N(R ａ )-,or -S (O) ２ - and R ａ Each occurrence of ３ and R １ is H, deuterium, halogen, -OH, C １－４ Alkyl, C １－４ Haloalkyl, C １－４ Alkoxy, or C １－４ is haloalkoxy; R ２ Each occurrence of is H, deuterium, halogen, -OH, CN, C １－８ Alkyl, C ２－８ Alkenyl, C ２－８ Alkynyl, C １－８ Alkoxy, -(CH ２ ) ｎ OR ６ , -(CH ２ ) ｎ S.R. ６ , -(CH ２ ) ｎ C(O)R ６ , -(CH ２ ) ｎ C(O)OR ６ , -(CH ２ ) ｎ S(O) ｍ R ６ 、-(CH ２ ) ｎ NR ７ R ８ 、-(CH ２ ) ｎ C(O)NR ７ R ８ 、-(CH ２ ) ｎ NR ７ C(O)R ６ 、 - (CH ２ ) ｎ N.R. ７ S (O) ｍ R ６ , C ３－８ cycloalkyl, 3- to 10-membered heterocyclyl, 6- to 14-membered aryl, 5- to 14-membered heteroaryl; R ２ The alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocyclyl, aryl, or heteroaryl represented by the formula (I) can each be selected from deuterium, halogen, CN, -OH, C １－８ Alkyl, C １－８ Haloalkyl, C １－８ Alkoxy, and NR ７ R ８ or When ring B is a 3- to 10-membered heterocyclyl, two R ２ Deuterium, halogens, CN, -OH, C １－８ Alkyl, C １－８ Haloalkyl, C １－８ Alkoxy, C １－８ Haloalkoxy, and NR ７ R ８ Optionally substituted with one or more groups selected from C ３－６ may form a cycloalkyl or a 3- to 6-membered heterocyclyl; R ３ Each occurrence of is independently H, deuterium, C １－８ Alkyl, C ２－８ Alkenyl, C ２－８ Alkynyl, C ３－８ cycloalkyl, or 3- to 10-membered heterocyclyl; R ３ The C represented by １－８ Alkyl, C ２－８ Alkenyl, C ２－８ Alkynyl, C ３－８ Cycloalkyl or 3-10 membered heterocyclyl is selected from the group consisting of deuterium, halogen, CN, —OH, C １－８ Alkyl, and C １－８ haloalkyl; R ４ Each occurrence of is independently H, deuterium, halogen, CN, C １－８ Alkyl, C ２－８ Alkenyl, C ２－８ Alkynyl, C １－８ Alkoxy, C(O)C １－８ Alkyl, C ３－８ cycloalkyl, or 3- to 10-membered heterocyclyl; R ４ or R ４ In the group represented by １－８ Alkyl, C ２－８ Alkenyl, C ２－８ Alkynyl, C １－８ Alkoxy, C ３－８ Cycloalkyl or 3- to 10-membered heterocyclyl is independently selected from deuterium, halogen, —OH, C １－８ Alkyl and C １－８ haloalkyl; or Two R's bonded to the same ring atom of ring A ４ The group is C ３－６ cycloalkyl or 3-6 membered heterocyclyl, each of which is selected from the group consisting of deuterium, halogen, CN, —OH, C １－８ Alkyl, C １－８ Haloalkyl, C １－８ Alkoxy, C １－８ Haloalkoxy, and NR ７ R ８ and optionally substituted with one or more groups selected from R ５ Each occurrence of is H, deuterium, halogen, -OH, CN, C １－４ Alkyl, C １－４ Haloalkyl, C １－４ Alkoxy, C １－４ Haloalkoxy, C(O)C １－４ alkyl, or 3- to 6-membered heterocyclyl; R ６ Each occurrence of １－８ Alkyl, C ２－８ Alkenyl, C ２－８ Alkynyl, C ３－８ cycloalkyl, 3- to 10-membered heterocyclyl, 6- to 14-membered aryl, 5- to 14-membered heteroaryl; R ６ The C represented by １－８ Alkyl, C ２－８ Alkenyl, C ２－８ Alkynyl, C ３－８ Cycloalkyl, The 3- to 10-membered heterocyclyl, the 6- to 14-membered aryl, and the 5- to 14-membered heteroaryl are each independently selected from halogen, CN, —OH, C １－４ Alkyl, C １－４ Haloalkyl, C １－４ Alkoxy, and NR ７ R ８ and optionally substituted with one or more groups selected from R ７ and R ８ Each occurrence of １－４ alkyl or cyclopropyl; R １２ Each occurrence of is H, deuterium, halogen, -OH, CN, NH ２ , C １－８ Alkyl, C ２－８ Alkenyl, C ２－８ Alkynyl, C １－８ Alkoxy, C ３－８ cycloalkyl, or 3- to 10-membered heterocyclyl; R １２ The C represented by １－８ Alkyl, C ２－８ Alkenyl, C ２－８ Alkynyl, C １－８ Alkoxy, C ３－８ cycloalkyl, or The 3- to 10-membered heterocyclyl is selected from halogen, CN, —OH, C １－４ Alkyl, C １－４ Haloalkyl, and C １－４ Optionally substituted with one or more groups selected from alkoxy; q is 0, 1, or 2; n is 0, 1, 2, 3, 4, or 5; m is 0, 1, or 2), or a pharma- ceutically acceptable salt, or stereoisomer thereof.

2. Ring C is 【Chemistry 3】 2. The compound of claim 1, wherein:

3. Ring A is 【Chemistry 4】 and During the ceremony, R ３ Each occurrence of １－４ C optionally substituted with alkyl or -OH ３－６ is cycloalkyl, R ４ Each occurrence of is optionally substituted with H, deuterium, halogen, fluoro. １－４ Alkyl, C ２－４ alkenyl, C optionally substituted with methyl ３－６ cycloalkyl, or 3- to 6-membered heterocyclyl, or two R ４ The group is C ３－６ cycloalkyl or 3-6 membered heterocyclyl, each of which is selected from halogen, CN, —OH, C １－２ Alkyl, C １－２ Alkoxy, and NR ７ R ８ and optionally substituted with one or more groups selected from R ５ is H, deuterium, halogen, CN, -OH, C １－４ Alkyl, C １－４ Alkoxy, or C １－４ 3. The compound of claim 1 or 2, or a pharma- ceutically acceptable salt, or stereoisomer thereof, which is haloalkoxy.

4. The compound has structural formulas (II-A) to (II-J): 【Chemistry 5】 or a pharma- ceutically acceptable salt or stereoisomer thereof, wherein R １２ teeth , H, F, Cl, CH ３ , or C.F. ３ and k is 0, 1, or 2.

5. L is a bond, -(CH ２ ) -, -O(CH ２ )-, -C(=O)-, or -S(O) ２ The compound according to any one of claims 1 to 4, or a pharma- ceutically acceptable salt or stereoisomer thereof, wherein

6. Ring B is one or two R ２ 6. The compound according to claim 1, or a pharma- ceutically acceptable salt or stereoisomer thereof, which is a 4-10 membered heterocyclyl or a 5-6 membered monocyclic heteroaryl optionally substituted with a group.

7. R ２ Each occurrence of is H, halogen, CN, -OH, C １－４ Alkyl, C １－４ Haloalkyl, C １－４ Hydroxyalkyl, -(CH ２ ) ｎ OR ６ , -(CH ２ ) ｎ C(O)R ６ , -(CH ２ ) ｎ C(O)OR ６ , -(CH ２ ) ｎ S (O) ２ R ６ , -(CH ２ ) ｎ NR ７ R ８ ,-(EH ２ ) ｎ C(O)NR ７ R ８ ,-(EH ２ ) ｎ C(O)NHR ７ ,-(EH ２ ) ｎ NR ７ C(O)R ６ 、 - (CH ２ ) ｎ N.R. ７ S (O) ２ R ６ , C ３－８ cycloalkyl, 3- to 6-membered heterocyclyl, phenyl, or 5-6 membered heteroaryl; or Two Rs bonded to the same ring atom of ring B ２ is halogen, -OH, C １－２ Alkyl, C １－２ Haloalkyl, C １－２ Alkoxy, C １－２ Haloalkoxy, and NR ７ R ８ and forming a 3-6 heterocyclyl (when Ring B is a 3-10 membered heterocyclyl) optionally substituted with one or more groups selected from R ６ Each occurrence of １－４ Alkyl, C ３－６ Cycloalkyl, 3-7 membered heterocyclyl, phenyl, 5-6 membered heteroaryl; R ６ C represented by １－４ Alkyl, C ３－６ Cycloalkyl, 3- to 7-membered heterocyclyl, phenyl, and 5- to 6-membered heteroaryl are each independently selected from halogen, CN, —OH, C １－４ Alkyl, C １－４ Haloalkyl, C １－４ Alkoxy, or NR ７ R ８ and optionally substituted with 7. The compound of any one of claims 1 to 6, or a pharma- ceutically acceptable salt, or stereoisomer thereof, wherein n is 0, 1, or 2.

8. R １ is H, F, Cl, or CH ３ 8. The compound according to any one of claims 1 to 7, or a pharma- ceutically acceptable salt or stereoisomer thereof,

9. Ring B 【Chemistry 6】 each of which contains one or two R ２ 9. The compound of any one of claims 1 to 8, or a pharma- ceutically acceptable salt, or stereoisomer thereof, optionally substituted with a group.

10. R ２ Each occurrence of is H, halogen, CN, -OH, C １－４ Alkyl, C １－４ Haloalkyl, C １－４ Hydroxyalkyl, -(CH ２ ) ｎ S (O) ２ C １－４ Alkyl, -(CH ２ ) ｎ N.R. ７ R ８ , C ３－４ cycloalkyl, or 3- to 6-membered heterocyclyl, where n is 0, 1, or 2; When ring B is a 4- to 7-membered heterocyclyl, two R ２ The compound according to any one of claims 1 to 9, or a pharma- ceutically acceptable salt, or stereoisomer thereof, wherein: forms a 3-6 heterocyclyl.

11. Ring A is 【Chemistry 7】 11. The compound according to any one of claims 1 to 10, or a pharma- ceutically acceptable salt or stereoisomer thereof,

12. R １ is H or F; or a compound according to any one of claims 1 to 11, A pharma- ceutically acceptable salt, or stereoisomer.

13. R ３ Each occurrence of １－３ C optionally substituted with alkyl or -OH ３－６ is cycloalkyl, R ４ Each occurrence of １－３ Alkyl, C ２－４ alkenyl, cyclopentyl, tetrahydro-2H-pyranyl, 3,6-dihydro-2H-pyranyl, R ５ Each occurrence of is H, F, CN, methoxy, OCHF ２ 13. The compound according to any one of claims 1 to 12, or a pharma- ceutically acceptable salt, or stereoisomer thereof, wherein:

14. 14. The compound of any one of claims 1 to 13, or a pharma- ceutically acceptable salt, or stereoisomer thereof, wherein L is a bond.

15. Ring B 【Chemistry 8】 each of which contains one or two R ２ 15. The compound of any one of claims 1 to 14, or a pharma- ceutically acceptable salt, or stereoisomer thereof, optionally substituted with a group.

16. R ２ Each occurrence of is H, halogen, CN, C １－３ Alkyl, C １－３ Haloalkyl, C １－３ Hydroxyalkyl, NH ２ , N(CH ３ ) ２ , NHcyclopropyl, -(CH ２ ) ｎ S (O) ２ C １－３ alkyl, cyclopropyl, azetidinyl optionally substituted with F, oxetanyl, morpholinyl, piperidinyl, tetrahydro-2H-pyranyl, or When ring B is piperidinyl, two R ２ is 2,5-pyrrolidinedionyl, or forming 2-pyrrolidonyl, 16. The compound of any one of claims 1 to 15, or a pharma- ceutically acceptable salt, or stereoisomer thereof, wherein n is 0, 1, or 2.

17. Ring A is 【Chemistry 9】 17. The compound according to any one of claims 1 to 16, or a pharma- ceutically acceptable salt, or stereoisomer thereof, wherein:

18. R ３ Each occurrence of １－３ is alkyl, R ４ Each occurrence of １－３ is alkyl, R ５ 18. The compound of any one of claims 1 to 17, or a pharma- ceutically acceptable salt, or stereoisomer thereof, wherein each occurrence of is H, F, or OMe.

19. R １ is H, or a pharma- ceutically acceptable salt, or stereoisomer thereof.

20. 20. The compound of any one of claims 1 to 19, or a pharma- ceutically acceptable salt, or stereoisomer thereof, wherein Ring C is unsubstituted.

21. Ring B 【Chemistry 10】 each of which contains one or two R ２ 21. The compound of any one of claims 1 to 20, or a pharma- ceutically acceptable salt, or stereoisomer thereof, optionally substituted with a group.

22. R ２ Each occurrence of is H, halogen, CN, C １－３ Alkyl, C １－３ Haloalkyl, C １－３ Hydroxyalkyl, NH ２ , N(CH ３ ) ２ 22. The compound of any one of claims 1 to 21, or a pharma- ceutically acceptable salt, or stereoisomer thereof, wherein R is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24,

23. 2. The compound of claim 1, or a pharma- ceutically acceptable salt or stereoisomer thereof, wherein the compound is a compound listed in the Examples.

24. 24. A pharmaceutical composition comprising an effective amount of a compound according to any one of claims 1 to 23, or a pharma- ceutically acceptable salt, or stereoisomer thereof, and a pharma- ceutically acceptable carrier.

25. A method of treating cancer, comprising administering to a subject in need thereof an effective amount of a compound according to any one of claims 1 to 23, or a pharma- ceutically acceptable salt or stereoisomer thereof, wherein the cancer is selected from the group consisting of colorectal cancer, breast cancer, lung cancer, prostate cancer, glioblastoma, mantle cell lymphoma, chronic myelogenous leukemia, and acute myelogenous leukemia. The method to be selected.

26. 26. A method of treating cancer by inhibiting a cyclin dependent kinase (CDK), said method comprising administering to a subject in need thereof an effective amount of a compound according to any one of claims 1 to 23.

27. 27. The method of claim 26, wherein the cancer is cancer of the bladder, breast, colon, kidney, epidermis, liver, lung, esophagus, gallbladder, ovary, pancreas, stomach, cervix, thyroid, nose, head and neck, prostate, or skin, hematopoietic tumors of the lymphoid system, hematopoietic tumors of the myeloid system, follicular thyroid cancer, tumors of mesenchymal origin, tumors of the central or peripheral nervous system, melanoma, seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum, keratoma, follicular thyroid cancer, or Kaposi's sarcoma.

28. 28. The method of claim 27, wherein the lymphatic hematopoietic tumor is leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, B-cell lymphoma, T-cell lymphoma, multiple myeloma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma, or Burkitt's lymphoma.

29. The cancer is ＋ Breast cancer, or hormone receptor (HR) positive (e.g., estrogen receptor positive (ER) ＋ ), progesterone receptor positive (PR ＋ ), or E.R. ＋ PR ＋ ), HER2 / neu negative cancer.

30. 30. The method of claim 29, wherein the cancer is advanced or metastatic or recurrent breast cancer.

31. 31. The method of claim 30, wherein the breast cancer is in an adult or postmenopausal woman.

32. 32. The method of any one of claims 29-31, further comprising administering a second agent selected from an aromatase inhibitor, a selective estrogen receptor modulator (SERM), a pure anti-estrogen with no estrogen agonist activity, a compound that temporarily suppresses ovarian function (e.g., estrogen and / or progesterone production), such as a gonadotropin-releasing hormone (GnRH) agonist or a luteinizing hormone-releasing hormone (LH-RH) agonist, a compound that inhibits CCYP3A4, or a monoclonal antibody against IGF-1 / IGF-2 or an antigen-binding fragment thereof.

33. 33. The method of any one of claims 25 to 32, further comprising administering an immune checkpoint inhibitor (such as a PD-1 inhibitor, a PD-L1 inhibitor, or a CTLA-4 inhibitor), a receptor Tyr kinase inhibitor, and / or an antagonist of a hormone receptor (such as an estrogen receptor).

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