Specific chemical composition and method of use thereof

Abstract

The present disclosure provides a CDK9 inhibitor.Also provided is a method for treating disease or disorder, comprising administering one of the CDK9 inhibitors disclosed herein to the subject in need of treatment.In some embodiments, the disease or disorder to be treated is cancer.In some embodiments, the disease or disorder is liver cancer.

Description

Technical Field

[0001] Cross-reference This application claims the benefit of U.S. Provisional Patent Application No. 63 / 025,474, filed May 15, 2020, which is hereby incorporated by reference herein in its entirety.

Background Art

[0002] Although there have been advances, cancer treatment remains relatively difficult. Systemic treatments such as chemotherapy are toxic and can cause harmful side effects to patients. CDK9 inhibitors have been regarded as promising small molecule cancer therapeutics, but their potential usefulness has been limited because of insufficient targeting to cancer tissues, resulting in peripheral exposure.

Summary of the Invention

[0003] Therefore, there is a need for the development of CDK9 inhibitors as small molecule cancer therapeutics with improved targeting.

[0004] In one aspect, a compound of formula (I):

[0005]

Chemical Formula

[0006] In some embodiments, ring A is C 3-6 Selected from cycloalkyl and 3-10 membered heterocycloalkyl. In some embodiments, ring A is C 3-6 It is a cycloalkyl. In some embodiments, ring A is

[0007] [ka] It is selected from the group consisting of the following.

[0008] In some embodiments, ring A is

[0009] [ka] It is selected from the group consisting of the following.

[0010] In some embodiments, ring A is

[0011] [ka] It is selected from the group consisting of the following.

[0012] In some embodiments, ring A is

[0013] [ka] It is selected from the group consisting of the following.

[0014] In some embodiments, X 1 , X 2 , X 3 , and X 4 One of them is N. In some embodiments, X 1 , X 2 , X 3 , and X 4 None of them are N.

[0015] In some embodiments, R 11, R 12 , R 13 , and R 14 These are H, Halo, -CN, and -OR, respectively, independently. 18 , -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 , -C(O)R 18 , -C(O)OR 18 , -NR 19 C(O)R 18 , C1-6 alkyl, and C 1-6 Selected from haloalkyls. In some embodiments, R 11 , R 12 , R 13 , and R 14 These are H, Halo, -CN, and -OR, respectively, independently. 18 , -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 , -C(O)R 18 , -C(O)OR 18 , and -NR 19 C(O)R 18 Selected from. In some embodiments, R 11 , R 12 , R 13 , and R 14 These are H, Halo, -CN, and -OR, respectively, independently. 18 , and -NR 16 R 17 Selected from. In some embodiments, R 11 is chloroform, R 12 , R 13 , and R 14 These are H.

[0016] In some embodiments, R 1 H, C1-6 alkyl, C 1-6 Haloalkyl and C 3-6 Selected from cycloalkyl. In some embodiments, R 1is Me. In some embodiments, R 1 H is H.

[0017] In some embodiments, R 2 is -CN, -OR 18 -SOR 15 , -SO2R 15 , -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 , -C(O)R 18 , -C(O)OR 18 , -NR 19 C(O)R 18 , -NR 19 C(O)NR 16 R 17 , -NR 19 SO2R 15 , -NR 19 SO2NR 16 R 17 , C1-6 alkyl, C 1-6 Haloalkyl and C 3-6 Selected from cycloalkyl. In some embodiments, R 2 R is selected from halo, -CN, -OH, -OMe, -OEt, -NH2, -NHMe, -NMe2, Me, Et, n-Pr, i-Pr, -CF3, and cyclopropyl. In some embodiments, R 2 This is Me.

[0018] In some embodiments, R 3 H, C1-6 alkyl, C 1-6 Haloalkyl, C 3-6 Selected from cycloalkyls and 3- to 10-membered heterocycloalkyls. In some embodiments, R 3 R is selected from H, Me, Et, -CF3, and cyclopropyl. In some embodiments, R 3 H is H.

[0019] In some embodiments, R 2 is Me, and R3 H is H.

[0020] In some embodiments, R 4 C 6-10 Aryl, 6-10 member heteroaryl, -O(C 0-4 Alkyl)C 3-6 Cycloalkyl, -O(C 0-4 Alkyl)(3-10 member heterocycloalkyl), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl) (6-10 member heteroaryl), -O(C 0-4 Alkyl)C(O)OR 18 , -O(C 0-4 Alkyl)C(O)NR 19 SO2R 15 , -O(C 0-4 Alkyl)SO2NR 19 C(O)R 18 , -O(C 3-6 Cycloalkyl)C 3-6 Cycloalkyl, -O(C 3-6 Cycloalkyl) (3-10 member heterocycloalkyl), -O(C 3-6 Cycloalkyl)C 6-10 Aryl, -O(C 3-6 Cycloalkyl) (6-10 member heteroaryl), -O(C 3-6 Cycloalkyl)C(O)OR 18 ,-(C 1-4 Alkyl)C 3-6 Cycloalkyl, -(C 1-4 Alkyl) (3-10 member heterocycloalkyl), -(C 1-4 Alkyl)C 6-10 Ariel, -(C 1-4 Alkyl) (6-10 member heteroaryl), and -(C 1-4 Alkyl)C(O)OR 18 Selected from, each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently C 1-4 Alkyl, oxo, halo, -OR 18 -CN, -NR 16 R 17-C(O)NR 16 R 17 -SO2NR 16 R 17 , -C(O)R 18 , -C(O)OR 18 ,-(C 1-4 Alkyl)OC(O)(C 1-4 Alkyl), -(C 1-4 Alkyl)OC(O)OR 18 , -NR 19 C(O)R 18 , -NR 19 C(O)NR 16 R 17 , -NR 19 SO2R 15 , and -NR 19 SO2NR 16 R 17 Optionally substituted with one or more substituents selected from, R 4’ and R 4” These are, independently, H, C1-6 alkyl, and C 1-6 Haloalkyl and C 3-6 Selected from cycloalkyls, where each alkyl and cycloalkyl is independently halo, -OR 18 -CN and -NR 16 R 17 Optionally substituted with one or more substituents selected from, or R 3 H is, R 4 , R 4’ , and R 4” Together with the carbon atoms to which they are bonded, -C(O)R 18 Alternatively, they form a 6-10 member heteroaryl group.

[0021] In some embodiments, R 4 These are 6-10 member heteroaryl compounds, -O(C 0-4 Alkyl)(3-10 member heterocycloalkyl), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4Alkyl) (6-10 member heteroaryl), -O(C 0-4 Alkyl)C(O)OR 18 , -O(C 0-4 Alkyl)C(O)NR 19 SO2R 15 , -O(C 0-4 Alkyl)SO2NR 19 C(O)R 18 , -O(C 3-6 Cycloalkyl) (6-10 member heteroaryl), -O(C 3-6 Cycloalkyl)C(O)OR 18 ,-(C 1-4 Alkyl) (6-10 member heteroaryl), and -(C 1-4 Alkyl)C(O)OR 18 Selected from, where each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently C 1-4 Alkyl, oxo, halo, -OR 18 -CN, -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 ,-C(O)OR 18 ,-(C 1-4 Alkyl)OC(O)(C 1-4 Alkyl), -(C 1-4 Alkyl)OC(O)OR 18 , -NR 19 C(O)R 18 , -NR 19 C(O)NR 16 R 17 , -NR 19 SO2R 15 , and -NR 19 SO2NR 16 R 17 Optionally substituted with one or more substituents selected from, R 4’ and R 4” Both are H, or R 3 H is, R 4 , R 4’ , and R4” Together with the carbon atoms to which they are bonded, -C(O)R 18 Alternatively, they form a 6-10 member heteroaryl group.

[0022] In some embodiments, R 4 These are 6-10 member heteroaryl compounds, -O(C 0-4 Alkyl)(3-10 member heterocycloalkyl), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl) (6-10 member heteroaryl), -O(C 0-4 Alkyl)C(O)OR 18 , -O(C 0-4 Alkyl)C(O)NR 19 SO2R 15 , -O(C 0-4 Alkyl)SO2NR 19 C(O)R 18 , -O(C 3-6 Cycloalkyl) (6-10 member heteroaryl), -O(C 3-6 Cycloalkyl)C(O)OR 18 ,-(C 1-4 Alkyl) (6-10 member heteroaryl), and -(C 1-4 Alkyl)C(O)OR 18 Selected from, where each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently C 1-4 Alkyl, oxo, halo, -OR 18 -CN, -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 ,-C(O)OR 18 ,-(C 1-4 Alkyl)OC(O)(C 1-4 Alkyl), -(C 1-4 Alkyl)OC(O)OR 18 , -NR 19 C(O)R 18 , -NR 19 C(O)NR 16 R17 , -NR 19 SO2R 15 , and -NR 19 SO2NR 16 R 17 Optionally substituted with one or more substituents selected from, R 4’ and R 4” Both are H.

[0023] In some embodiments, R 3 H is, R 4 , R 4’ , and R 4” Together with the carbon atoms to which they are bonded, -C(O)R 18 Alternatively, they form a 6-10 member heteroaryl group.

[0024] In some embodiments, R 5 H, C1-6 alkyl, C 1-6 Haloalkyl and C 3-6 Selected from cycloalkyl. In some embodiments, R 5 is Me. In some embodiments, R 5 H is H.

[0025] In some embodiments, R 6 and R 7 These are H and -(C) respectively, independently. 1-4 Alkyl)C 3-6 Cycloalkyl, -(C 1-4 Alkyl) (3-10 member heterocycloalkyl), -(C 1-4 Alkyl)C 6-10 Aryl, and -(C 1-4 Selected from alkyl (6-10 member heteroaryl), where each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently oxo, halo, -OR 18 , C1-4 alkyl, C1-4 alkoxy, C 1-4 Heteroalkyl, C 1-4Haloalkyl, -CN, and -NR 20 R 21 Optionally substituted with one or more substituents selected from, or R 6 and R 7 These, together with the nitrogen atom to which they are bonded, form oxo, halo, -OR 18 , C1-4 alkyl, C1-4 alkoxy, C 1-4 Heteroalkyl, C 1-4 Haloalkyl, -CN, and -NR 20 R 21 It forms a 3- to 10-membered heterocycloalkyl group, which is optionally substituted with one or more substituents selected from the following.

[0026] In some embodiments, R 6 and R 7 These are H and -(C) respectively, independently. 1-4 Selected from alkyl (3-10 member heterocycloalkyl), where each alkyl and heterocycloalkyl is independently halo, -OR 18 -CN and -NR 20 R 21 It is optionally substituted with one or more substituents selected from the following.

[0027] In some embodiments, R 6 and R 7 One of them is H, and the other is

[0028] [ka] That is the case.

[0029] In some embodiments, R 8 and R 9 These are H, Halo, -CN, and -OR, respectively, independently. 18 , -SO2R 15 , -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR16 R 17 , -C(O)OR 18 , -NR 19 C(O)R 18 , -NR 19 SO2R 15 Selected from. In some embodiments, R 8 and R 9 These are H, Halo, -CN, and -OR, respectively, independently. 18 , and -NR 16 R 17 Selected from. In some embodiments, R 8 and R 9 Both are H.

[0030] In some embodiments, n is 0, 1, or 2. In some embodiments, n is 0.

[0031] In some embodiments, the compound of formula (I) is formula (IA):

[0032] [ka] It is represented by [this].

[0033] In some embodiments, the compound of formula (I) is formula (IB):

[0034] [ka] It is represented by [this].

[0035] In some embodiments, the compound of formula (I) is formula (IC), formula (ID), formula (IE), or formula (IF):

[0036] [ka] It is represented by [this].

[0037] In some embodiments, the compound is

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[0038]

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[0039]

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[0040]

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[0041]

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[0042]

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[0043]

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[0044]

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[0045]

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[0046]

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[0047]

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[0048] In another embodiment, a pharmaceutical composition comprising a compound described herein or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient is provided herein.

[0049] In another embodiment, a method for treating a disease or disorder in a patient in need thereof is provided herein, comprising administering a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, to a patient in need thereof.

[0050] In some embodiments, the disease or disorder is cancer. In some embodiments, the cancer is selected from leukemia, breast cancer, prostate cancer, ovarian cancer, colon cancer, cervical cancer, lung cancer, lymphoma, and liver cancer. In some embodiments, the cancer is liver cancer.

[0051] Reference All publications, patents, and patent applications referenced herein are incorporated herein by reference to the same extent as they are specifically and individually indicated for each individual publication, patent, or patent application.

[0052] The novel features of the present invention are described in detail in the appended claims. The features and advantages of the present invention will be better understood by referring to the following detailed description and accompanying drawings illustrating exemplary embodiments in which the principles of the present invention are utilized. [Brief explanation of the drawing]

[0053] [Figure 1] This graph shows the ratio of compound concentrations in the liver and blood of CD-1 mice after a single oral administration of a 5 mg / kg compound suspension. [Figure 2]This graph shows the ratio of compound concentrations in blood collected from the jugular vein and portal vein of Sprague-Dawley (SD) rats after a single oral administration of a 5 mg / kg compound suspension. [Figure 3] This graph shows the mean body weight change (relative to day 1) of BALB / c nude mice treated with the vehicle and compound. [Modes for carrying out the invention]

[0054] definition Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art in the field to which this disclosure pertains.

[0055] As used herein, unless the context explicitly states otherwise, the singular forms “a,” “an,” and “the” refer to multiple objects.

[0056] When used in combination with chemical moieties such as alkyl, alkenyl, or alkynyl, "C x-y The term "C" is intended to encompass groups containing x to y carbon atoms in the chain. For example, "C 1-6 The term "alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups, including linear and branched alkyl groups containing 1 to 6 carbon atoms. -C x-y The term alkylene- refers to a substituted or unsubstituted alkylene chain having x to y carbon atoms in the alkylene chain. For example, -C 1-6 The alkylene- may be selected from methylene, ethylene, propylene, butylene, pentylene, and hexylene, and any of these may be substituted.

[0057] "Alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups, including linear and branched alkyl groups. Alkyl groups consist of 1 to 12 carbon atoms (for example, C 1-12Alkyl), for example, 1 to 8 carbon atoms (C 1-8 Alkyl) or 1 to 6 carbon atoms (C 1-6 The molecule may contain alkyl groups. Exemplary alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, hexyl, septyl, octyl, nonyl, and decyl. The alkyl group is bonded to the rest of the molecule by a single bond. Unless otherwise specifically mentioned herein, the alkyl group may be substituted with one or more substituents, such as substituents described herein.

[0058] "Haloalkyl" refers to an alkyl group substituted with one or more halogens. Examples of haloalkyl groups include trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, and 1,2-dibromoethyl.

[0059] "Alkenyl" refers to a substituted or unsubstituted hydrocarbon group containing a linear or branched alkenyl group with at least one double bond. The alkenyl group may contain 2 to 12 carbon atoms (for example, C 2-12 Alkenyl). Exemplary alkenyl groups include ethenyl (i.e., vinyl), propa-1-enyl, buta-1-enyl, penta-1-enyl, penta-1,4-dienyl, and the like. Unless otherwise specifically mentioned herein, the alkenyl group may be substituted with one or more substituents, such as those described herein.

[0060] "Alkynyl" refers to a substituted or unsubstituted hydrocarbon group containing a linear or branched alkynyl group with at least one triple bond. The alkynyl group may contain 2 to 12 carbon atoms (for example, C 2-12Alkynyl groups. Exemplary alkynyl groups include ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like. Unless otherwise specifically mentioned herein, the alkynyl group may be substituted with one or more substituents, such as those described herein.

[0061] "Heteroalkyl," "heteroalkenyl," and "heteroalkynyl" refer to substituted or unsubstituted alkyl, alkenyl, and alkynyl groups, respectively, having one or more skeletal chain atoms selected from atoms other than carbon. Examples of skeletal chain atoms selected from atoms other than carbon include, for example, O, N, P, Si, S, or combinations thereof, where nitrogen, phosphorus, and sulfur atoms may be optionally oxidized, and the nitrogen heteroatom may be optionally quaternized. Where a numerical range is given, it refers to the overall chain length. For example, a 3- to 8-membered heteroalkyl has a chain length of 3 to 8 atoms. The rest of the molecule may be connected either through heteroatoms or carbon in the heteroalkyl chain, heteroalkenyl chain, or heteroalkynyl chain. Unless otherwise specifically mentioned herein, the heteroalkyl, heteroalkenyl, or heteroalkynyl group may be substituted with one or more substituents, such as those described herein.

[0062] "Aryl" refers to an aromatic ring in which each of the atoms forming the ring is a carbon atom. The aryl group may be substituted. Examples of aryl groups include, but are not limited to, phenyl and naphthyl. In some embodiments, the aryl is phenyl. Depending on the structure, the aryl group may be a monoradical or a diradical (i.e., an arylene group). Unless otherwise specifically referred to herein, the term "aryl" or the prefix "ar-" (for example, in "aralkyl") is intended to encompass substituted aryl radicals.

[0063] "Heteroaryl" refers to a 3- to 12-membered aromatic ring containing at least one heteroatom, where each heteroatom may be independently selected from N, O, and S. As used herein, heteroaryl rings may be selected from monocyclic or bicyclic, fused or bridging ring systems, where at least one of the rings in the ring system contains an aromatic, i.e., a cyclic, delocalized (4n+2) π-electron system according to Hückel's theory. Heteroatoms in heteroaryls are optionally oxidized. If present, one or more nitrogen atoms may be quaternized. Heteroaryls may be bonded to the rest of the molecule through any atom of the heteroaryl, such as the carbon or nitrogen atoms of the heteroaryl, as long as their valence allows. Examples of heteroaryls, though not limited to them, include azepinyl, acridinyl, benzimidazolyl, benzoindolyl, 1,3-benzodioxolyl, benzofuranil, benzoxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl, 1,4-benzodioxanil, benzonaphthofuranil, benzoxazolyl, benzodioxolyl, benzodioxynil, benzopyranil, benzopyranonil, benzofuranil, benzothienyl (benzothiophenyl), benzothieno[3,2-d]pyrimidinil, benzotriazolyl, benzo[4,6 ]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, cyclopenta[d]pyrimidinyl, 6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl, 6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridadinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, furo[3,2-c]pyridinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl, 5,6,7,8,9,10-Hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl, indazolyl, indazolyl, isoindolyl, indolinyl, isoindolyl, isoquinolyl, indolidinyl, isoxazolyl, 5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthilidinyl, 1,6-naphthilidinolyl, oxadiazolyl, 2-o Xoazepinyl, oxazolyl, oxyranil, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxadinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl Dinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridadinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl, 6,7,8,9-tetrahydro-5H-cyclo Examples include hepta[4,5]thieno[2,3-d]pyrimidinyl, 5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pyrimidinyl, and thiophenyl (i.e., thienyl). Unless otherwise specifically mentioned herein, heteroaryls may be substituted with one or more substituents, such as those described herein.

[0064] The term "cycloalkyl" refers to monocyclic or polycyclic non-aromatic radicals in which each of the ring-forming atoms (i.e., skeletal atoms) is a carbon atom. In some embodiments, cycloalkyls are saturated or partially unsaturated. In some embodiments, cycloalkyls are spirocyclic or crosslinked compounds. In some embodiments, cycloalkyls are condensed with an aromatic ring (in which case the cycloalkyl is bonded through the carbon atoms of the non-aromatic ring). Examples of cycloalkyl groups include those having 3 to 10 ring atoms. Representative cycloalkyls, but are not limited to, include those having 3 to 10 carbon atoms, 3 to 8 carbon atoms, 3 to 6 carbon atoms, or 3 to 5 carbon atoms. Examples of monocyclic cycloalkyl radicals include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Examples of polycyclic radicals include adamantyl, 1,2-dihydronaphthalenyl, 1,4-dihydronaphthalenyl, tetralinyl, dekalinyl, 3,4-dihydronaphthalenyl-1(2H)-one, spiro[2.2]pentyl, norbornyl, and bicyclo[1.1.1]pentyl. Unless otherwise specifically mentioned herein, cycloalkyl groups may be optionally substituted.

[0065] The term "heterocycloalkyl" refers to a cycloalkyl group comprising at least one heteroatom selected from nitrogen, oxygen, and sulfur. Unless otherwise specifically mentioned herein, heterocycloalkyl radicals may be monocyclic or bicyclic ring systems, which may include fused ring systems (where the heterocycloalkyl is bonded through a non-aromatic ring atom when fused with an aryl or heteroaryl ring) or cross-linked ring systems. The nitrogen, carbon, or sulfur atom in the heterocycloalkyl radical may be optionally oxidized. The nitrogen atom may optionally be quaternized. Heterocycloalkyl radicals may be partially saturated or fully saturated. Examples of heterocycloalkyl radicals, though not limited to them, include dioxolanil, thienyl[1,3]dithianil, tetrahydroquinolyl, tetrahydroisoquinolyl, decahydroquinolyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianil, tetrahydropyranil, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Furthermore, the term heterocycloalkyl encompasses, but is not limited to, all cyclic carbohydrates, including monosaccharides, disaccharides, and oligosaccharides. Unless otherwise noted, heterocycloalkyls have 2 to 12 carbon atoms in the ring. When referring to the number of carbon atoms in a heterocycloalkyl, it should be understood that the number of carbon atoms in a heterocycloalkyl is not the same as the total number of atoms (including heteroatoms) that make up the heterocycloalkyl (i.e., the skeletal atoms of the heterocycloalkyl ring). Unless otherwise specifically noted herein, heterocycloalkyls may be optionally substituted.

[0066] The term “substituted” refers to a portion of a structure having a substituent that replaces a hydrogen on one or more carbons or heteroatoms. “Substituted” or “substituted with” will be understood to implicitly include the assumption that such substitution conforms to the acceptable valencies of the substituted atom and substituent, and that the substitution results in a stable compound that does not spontaneously deform, for example, by rearrangement, cyclization, or elimination. Where used herein, the term “substituted” is intended to encompass all acceptable substituents of an organic compound. In broad terms, acceptable substituents of an organic compound include acyclic and cyclic, branched and unbranched, carboncyclic and heterocyclic, aromatic and non-aromatic substituents. For a given organic compound, there may be one or more, identical or different, acceptable substituents. For the purposes of this disclosure, a heteroatom, such as nitrogen, may have any acceptable substituent of an organic compound described herein that satisfies the hydrogen substituent and / or the valency of the heteroatom. Substituents include any substituents described herein, such as halogens, hydroxyls, carbonyls (e.g., carboxyls, alkoxycarbonyls, formyls, or acyls), thiocarbonyls (e.g., thioesters, thioacetates, or thioformates), alkoxyls, phosphoryls, phosphates, phosphonates, phosphinates, aminos, amides, amidines, imines, cyanos, nitros, azides, sulfhydryls, alkylthios, sulfates, sulfonates, sulfamoyls, sulfonamides, sulfonyls, heterocyclyls, aralkyls, carbocyclics, heterocyclics, cycloalkyls, heterocycloalkyls, aromatics, and heteroaromatic moieties.

[0067] Those skilled in the art will understand that substituents may be substituted themselves, where appropriate. Unless specifically referred to as “unsubstituted,” references to chemical moieties herein are understood to include substituted variants. For example, references to “heteroaryl” groups or moieties implicitly include both substituted and unsubstituted variants.

[0068] When substituents are specified by their conventional chemical formulas and written from left to right, they equally encompass chemically identical substituents that would result from writing the structure from right to left; for example, -CH2O- is equivalent to -OCH2-.

[0069] "Optional" or "optionally" means that the event in the situation described afterward may or may not occur, and the description includes both cases in which the event or situation occurs and cases in which it does not occur. For example, "optionally substituted aryl" means that the aryl group may or may not be substituted, and the description includes both substituted and unsubstituted aryl groups.

[0070] The compounds of this disclosure also encompass crystalline and amorphous forms of those compounds, pharmaceutically acceptable salts, and active metabolites of those compounds having the same kind of activity, such as polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrous forms), conformational polymorphs, and amorphous forms of the compounds, as well as mixtures thereof.

[0071] The compounds described herein may exhibit their natural isotopic abundances, or one or more of their atoms may be artificially enriched with specific isotopes having the same atomic number but with atomic masses or mass numbers different from those primarily found in nature. All isotopic variants of the compounds disclosed herein, whether radioactive or not, are included within the scope of this disclosure. For example, hydrogen, 1 H (protium), 2 H (deuterium), and 3It has three naturally occurring isotopes called tritium (H). Protium is the most abundant hydrogen isotope in nature. Enriching with deuterium may yield certain therapeutic benefits, such as increased in vivo half-life and / or exposure, or may provide compounds useful for investigating drug excretion and metabolic pathways in vivo. Isotope-enriched compounds can be prepared by prior art well known to those skilled in the art.

[0072] "Isomers" are different compounds that have the same molecular formula. "Stereoisomers" are isomers that differ only in how their atoms are spatially arranged. "Enantiomers" are a pair of stereoisomers that are mirror images of each other and cannot be superimposed. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. The term "(±)" is used to indicate a racemic mixture where appropriate. "Diastereoisomers" are stereoisomers that have at least two chiral atoms but are not mirror images of each other. Absolute stereochemistry is determined according to the Kahn-Ingold-Prelogue-RS system. If a compound is a pure enantiomer, the stereochemistry of each chiral carbon can be determined as either R or S. Divided compounds whose absolute configuration is unknown can be indicated as (+) or (-) depending on the direction in which the plane polarization of the sodium D line wavelength is rotated (dextrorotatory or levorotatory). Certain compounds described herein contain one or more chiral centers, thus resulting in enantiomers, diastereomers, and other stereoisomers, where these chiral centers can be defined as (R)- or (S)- from the standpoint of absolute stereochemistry. The chemical components, pharmaceutical compositions, and methods described herein are intended to encompass all such possible stereoisomers, including racemic mixtures, optically pure forms, mixtures of diastereomers, and mixtures of intermediates. Optically active (R)- and (S)-isomers can be prepared using chiral synthons or chiral reagents, or they can be separated using prior art. The optical activity of the compounds can be analyzed by any suitable method, including, but not limited to, chiral chromatography and polarimetric analysis, to determine the degree of superiority of one stereoisomer over the other.

[0073] Chemical components containing carbon-carbon double bonds or carbon-nitrogen double bonds may exist in Z-isomer or E-isomer (or cis- or trans-isomer). Furthermore, some chemical components may exist in various tautomers. Unless otherwise specified, the chemical components described herein are intended to encompass all Z-isomers, E-isomers, and tautomers.

[0074] The isolation and purification of the chemical components and intermediates described herein can be achieved by any suitable separation or purification procedure, if desired, such as filtration, extraction, crystallization, column chromatography, thin-layer chromatography, or thick-layer chromatography, or a combination thereof. Specific examples of suitable separation and isolation procedures can be provided by referring to the examples described later herein. However, other equivalent separation or isolation procedures may also be used.

[0075] Where the stereochemistry is not specified, certain small molecules described herein include, but are not limited to, enantiomers and diastereomers and their isomers, including racemates, mixtures of enantiomers, mixtures of diastereomers, and other mixtures thereof, to the extent that a person skilled in the art can prepare them by ordinary experiments, where possible. In those situations, a single enantiomer or diastereomer, i.e., an optically active form, can be obtained by asymmetric synthesis or by the resolution of a racemate or mixture of diastereomers. The resolution of a racemate or mixture of diastereomers can be achieved, where possible, by conventional methods such as crystallization in the presence of a resolving agent, or by chromatography, for example, by a chiral high-pressure liquid chromatography (HPLC) column. Furthermore, a mixture of two enantiomers, enriched with one of the two, can be purified and recrystallized and / or tritulated to provide a further optically enriched form of the major enantiomer. In addition, such a particular small molecule also includes the Z and E (or cis and trans) forms of a particular small molecule having a carbon-carbon double bond or a carbon-nitrogen double bond. If a particular small molecule described herein exists in various tautomer forms, the term “a particular small molecule” is intended to encompass all tautomer forms of that particular small molecule.

[0076] The terms “salt” or “pharmaceutically acceptable salt” refer to salts derived from various organic and inorganic counterions known in the art. pharmaceutically acceptable acid addition salts can be formed from inorganic and organic acids. Examples of inorganic acids that can derivate salts include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Examples of organic acids that can derivate salts include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. pharmaceutically acceptable base addition salts can be formed from inorganic and organic bases. Examples of inorganic bases that can derivate salts include sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Examples of organic bases that can induce salt formation include primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, specifically, for example, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, pharmaceutically acceptable base addition salts are selected from ammonium salts, potassium salts, sodium salts, calcium salts, and magnesium salts.

[0077] As used herein, the terms “pharmaceutically acceptable excipient” or “pharmaceutically acceptable carrier” mean a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be “acceptable” in the sense that it is compatible with the other components of the formulation and is not harmful to the patient. Some examples of materials that can act as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose, and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) tragacanth powder; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository wax; (9) peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil. Examples include oils; (10) glycols such as propylene glycol; (11) polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer; and (21) other non-toxic, suitable substances used in pharmaceutical formulations.

[0078] The terms “effective dose” or “therapeutic effective dose” mean the amount of a compound described herein that is sufficient to have an effect on the intended use (including, but not limited to, the treatment of a disease), as defined below. The therapeutic effective dose may vary depending on the intended therapeutic use (in vivo) or the subject and disease state being treated, such as the subject’s weight and age, the severity of the disease state, the mode of administration, and similar factors, which can be readily determined by those skilled in the art. The term also applies to the dose that induces a specific response in target cells, such as a reduction in platelet adhesion and / or cell migration. The specific dose will vary depending on the particular compound chosen, the administration regimen to be followed, whether it is administered in combination with other compounds, the timing of administration, the tissue to which it is administered, and the physical delivery system to which it is carried.

[0079] As used herein, “treatment” or “to treat” refers to a method for obtaining beneficial or desired outcomes with respect to a disease, disorder, or medical condition, including but not limited to therapeutic and / or preventive benefits. A therapeutic benefit could be, for example, the eradication or remission of the underlying disorder being treated. A therapeutic benefit could also be the eradication or remission of one or more physiological symptoms associated with the underlying disorder, such that improvement is observed in the subject, even if the subject still suffers from the underlying disorder. In certain embodiments, for a preventive benefit, the composition is administered to a subject at risk of developing a particular disease, or to a subject reporting one or more physiological symptoms of the disease, even if the disease has not been diagnosed.

[0080] When this term is used herein, “therapeutic effect” includes the therapeutic and / or preventive benefits described above. Preventive effects include delaying or eliminating the onset of a disease or condition, delaying or eliminating the manifestation of symptoms of a disease or condition, delaying, halting, or reversing the progression of a disease or condition, or any combination thereof.

[0081] As used herein, the terms “co-administration,” “administered in combination with,” and their grammatical equivalents encompass the administration of two or more drugs to an animal, including a human, in which both drugs and / or their metabolites are present simultaneously in the subject. Co-administration includes simultaneous administration as separate compositions, administration at different times as separate compositions, or administration as a composition in which both drugs are present.

[0082] The terms “antagonist” and “inhibitor” are used interchangeably and refer to compounds that have the ability to inhibit the biological function (e.g., activity, expression, binding, protein-protein interaction) of a target protein or enzyme. Therefore, the terms “antagonist” and “inhibitor” are defined in the context of the biological role of the target protein. Preferred antagonists as used herein are compounds that specifically interact with (e.g., bind to) the target, but also inhibit the biological activity of the target protein by interacting with other members of the signaling pathway to which the target protein is a member. Preferred biological activities inhibited by antagonists are associated with tumor development, growth, or spread.

[0083] compound In one embodiment, formula (I):

[0084] [ka] Compounds thereof or pharmaceutically acceptable salts thereof are provided herein. During the ceremony, Ring A is C 3-6 Cycloalkyl, 3-10 member heterocycloalkyl, C 6-10 Selected from aryls and 6-10 membered heteroaryls, X 1 N and CR 11 Selected from, X 2 N and CR 12 Selected from, X3 N and CR 13 Selected from, X 4 N and CR 14 Selected from, R 1 H, C1-6 alkyl, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3-10 member heterocycloalkyl, C 6-10 Selected from aryls and 6-10 membered heteroaryls, where each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently oxo, halo, or -OR. 18 , C1-4 alkyl, C1-4 alkoxy, C 1-4 Heteroalkyl, C 1-4 Haloalkyl, -CN, and -NR 20 R 21 Optionally substituted with one or more substituents selected from, R 2 is -CN, -OR 18 -SOR 15 , -SO2R 15 , -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 , -C(O)R 18 , -C(O)OR 18 , -NR 19 C(O)R 18 , -NR 19 C(O)NR 16 R 17 , -NR 19 SO2R 15 , -NR 19 SO2NR 16 R 17 , C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3-10 member heterocycloalkyl, C 6-10Selected from aryls and 6-10 membered heteroaryls, where each alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently oxo, halo, -OR 18 , C1-4 alkyl, C1-4 alkoxy, C 1-4 Heteroalkyl, C 1-4 Haloalkyl, -CN, and -NR 20 R 21 Optionally substituted with one or more substituents selected from, R 3 H, Halo, -CN, -OR 18 -SOR 15 , -SO2R 15 , -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 , -C(O)R 18 , -C(O)OR 18 , -NR 19 C(O)R 18 , -NR 19 C(O)NR 16 R 17 , -NR 19 SO2R 15 , -NR 19 SO2NR 16 R 17 , C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3-10 member heterocycloalkyl, C 6-10 Selected from aryls and 6-10 membered heteroaryls, where each alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently oxo, halo, -OR 18 , C1-4 alkyl, C1-4 alkoxy, C 1-4 Heteroalkyl, C 1-4 Haloalkyl, -CN, and -NR 20 R 21Optionally substituted with one or more substituents selected from, R 4 C1-6 alkyl, C 2-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3-10 member heterocycloalkyl, C 6-10 Aryl, 6-10 member heteroaryl, -O(C 0-4 Alkyl)C 3-6 Cycloalkyl, -O(C 0-4 Alkyl)(3-10 member heterocycloalkyl), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl) (6-10 member heteroaryl), -O(C 0-4 Alkyl)C(O)OR 18 , -O(C 0-4 Alkyl)C(O)NR 19 SO2R 15 , -O(C 0-4 Alkyl)SO2NR 19 C(O)R 18 , -O(C 3-6 Cycloalkyl)C 3-6 Cycloalkyl, -O(C 3-6 Cycloalkyl) (3-10 member heterocycloalkyl), -O(C 3-6 Cycloalkyl)C 6-10 Aryl, -O(C 3-6 Cycloalkyl) (6-10 member heteroaryl), -O(C 3-6 Cycloalkyl)C(O)OR 18 ,-(C 1-4 Alkyl)C 3-6 Cycloalkyl, -(C 1-4 Alkyl) (3-10 member heterocycloalkyl), -(C 1-4 Alkyl)C 6-10 Ariel, -(C 1-4 Alkyl) (6-10 member heteroaryl), and -(C 1-4 Alkyl)C(O)OR 18 Selected from, where each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently C 1-4Alkyl, oxo, halo, -OR 18 -CN, -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 , -C(O)R 18 , -C(O)OR 18 ,-(C 1-4 Alkyl)OC(O)(C 1-4 Alkyl), -(C 1-4 Alkyl)OC(O)OR 18 , -NR 19 C(O)R 18 , -NR 19 C(O)NR 16 R 17 , -NR 19 SO2R 15 , and -NR 19 SO2NR 16 R 17 Optionally substituted with one or more substituents selected from, R 4’ and R 4” These are, independently, H, C1-6 alkyl, and C 2-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3-10 member heterocycloalkyl, C 6-10 Aryl, 6-10 member heteroaryl, -O(C 0-4 Alkyl)C 3-6 Cycloalkyl, -O(C 0-4 Alkyl)(3-10 member heterocycloalkyl), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl) (6-10 member heteroaryl), -O(C 0-4 Alkyl)C(O)OR 18 , -O(C 0-4 Alkyl)C(O)NR 19 SO2R 15 , -O(C 0-4 Alkyl)SO2NR 19 C(O)R 18 , -O(C 3-6 Cycloalkyl)C 3-6Cycloalkyl, -O(C 3-6 Cycloalkyl) (3-10 member heterocycloalkyl), -O(C 3-6 Cycloalkyl)C 6-10 Aryl, -O(C 3-6 Cycloalkyl) (6-10 member heteroaryl), -O(C 3-6 Cycloalkyl)C(O)OR 18 ,-(C 1-4 Alkyl)C 3-6 Cycloalkyl, -(C 1-4 Alkyl) (3-10 member heterocycloalkyl), -(C 1-4 Alkyl)C 6-10 Aryl, and -(C 1-4 Selected from alkyl (6-10 member heteroaryl), where each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently oxo, halo, -OR 18 -CN, -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 , -C(O)R 18 , -C(O)OR 18 ,-(C 1-4 Alkyl)OC(O)(C 1-4 Alkyl), -(C 1-4 Alkyl)OC(O)OR 18 , -NR 19 C(O)R 18 , -NR 19 C(O)NR 16 R 17 , -NR 19 SO2R 15 , and -NR 19 SO2NR 16 R 17 Optionally substituted with one or more substituents selected from, or R 3 H is, R 4 , R 4’ , and R 4” Together with the carbon atoms to which they are bonded, -C(O)R 18Alternatively, they form a 6-10 member heteroaryl compound. R 5 H, C1-6 alkyl, C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3-10 member heterocycloalkyl, C 6-10 Selected from aryls and 6-10 membered heteroaryls, where each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently oxo, halo, or -OR. 18 , C1-4 alkyl, C1-4 alkoxy, C 1-4 Heteroalkyl, C 1-4 Haloalkyl, -CN, and -NR 20 R 21 Optionally substituted with one or more substituents selected from, R 6 and R 7 These are, independently, H, C1-6 alkyl, and C 1-6 Heteroalkyl, C 1-6 Haloalkyl, C 3-6 Cycloalkyl, 3-10 member heterocycloalkyl, C 6-10 Aryl, 6-10 member heteroaryl, -(C 1-4 Alkyl)C 3-6 Cycloalkyl, -(C 1-4 Alkyl) (3-10 member heterocycloalkyl), -(C 1-4 Alkyl)C 6-10 Aryl, and -(C 1-4 Selected from alkyl (6-10 member heteroaryl), where each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently oxo, halo, -OR 18 , C1-4 alkyl, C1-4 alkoxy, C 1-4 Heteroalkyl, C 1-4 Haloalkyl, -CN, and -NR 20 R 21 Optionally substituted with one or more substituents selected from, or R 6 and R 7forms a 3- to 10-member heterocycloalkyl optionally substituted with one or more substituents selected from oxo, halo, -OR 18 , C1-4 alkyl, C1-4 alkoxy, C 1-4 heteroalkyl, C 1-4 haloalkyl, -CN, and -NR 20 R 21 and is optionally substituted with one or more substituents selected from oxo, halo, -OR R 8 and R 9 are each independently H, halo, -CN, -OR 18 , -SOR 15 , -SO2R 15 , -NR 16 R 17 , -C(O)NR 16 R 17 , -SO2NR 16 R 17 , -C(O)R 18 , -C(O)OR 18 , -NR 19 C(O)R 18 , -NR 19 C(O)NR 16 R 17 , -NR 19 SO2R 15 , -NR<照 19 SO2NR 16 R 17 , C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C 1-6 heteroalkyl, and C 1-6 haloalkyl, where each alkyl, alkenyl, and alkynyl is independently optionally substituted with one or more substituents selected from oxo, halo, -OR 18 , C1-4 alkyl, C1-4 alkoxy, C 1-4 heteroalkyl, C 1-4 haloalkyl, -CN, and -NR 20 R 21 and is optionally substituted with one or more substituents selected from oxo, halo, -OR each R 10 is independently halo, -CN, -OR 18 , -NR 16 R 17 , -C(O)NR 16 It should be noted that there may be an incorrect tag "照0000920" in the original text. It is recommended to double-check the original for accuracy.R 17 、 -SO₂NR 16 R 17 、 -C(O)R 18 、 -C(O)OR 18 、 -NR 19 C(O)R 18 、 C₁₋₆ alkyl, C 1-6 heteroalkyl, C 1-6 haloalkyl, C 3-6 cycloalkyl, 3 - 10 member heterocycloalkyl, where each alkyl, cycloalkyl, and heterocycloalkyl is independently optionally substituted with one or more substituents selected from oxo, halo, -OR 18 、 C₁₋₄ alkyl, C₁₋₄ alkoxy, C 1-4 heteroalkyl, C 1-4 haloalkyl, -CN, and -NR 20 R 21 from one or more substituents selected from, and is optionally substituted with R 11 、 R 12 、 R 13 、 and R 14 are each independently H, halo, -CN, -OR 18 、 -NR 16 R 17 、 -C(O)NR 16 R 17 、 -SO₂NR 16 R 17 、 -C(O)R 18 、 -C(O)OR 18 、 -NR 19 C(O)R 18 、 C₁₋₆ alkyl, C 1-6 heteroalkyl, C 1-6 haloalkyl, C 3-6 cycloalkyl, 3 - 10 member heterocycloalkyl, where each alkyl, cycloalkyl, and heterocycloalkyl is independently optionally substituted with one or more substituents selected from oxo, halo, -OR 18 、 C₁₋₄ alkyl, C₁₋₄ alkoxy, C 1-4 heteroalkyl, C 1-4 haloalkyl, -CN, and -NR 20 R 21 from one or more substituents selected from, and is optionally substituted with Each R 15 Independently, C 1-4 Alkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, 3-10 member heterocycloalkyl, C 6-10 Selected from aryls and 6-10 membered heteroaryls, Each R 16 and R 17 These are independently H, C1-4 alkyl, C 1-4 Heteroalkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, 3-10 member heterocycloalkyl, C 6-10 Selected from aryls, 6-10 member heteroaryls, or R 16 and R 17 These may form a 3- to 10-membered heterocycloalkyl group together with the nitrogen atom to which they are bonded. Each R 18 These are independently H, C1-4 alkyl, C 1-4 Heteroalkyl, C 1-4 Haloalkyl and C 3-6 Selected from cycloalkyl groups, Each R 19 These are independently H, C1-4 alkyl, C 1-4 Heteroalkyl, C 1-4 Haloalkyl and C 3-6 Selected from cycloalkyl groups, Each R 20 and R 21 These are independently H, C1-4 alkyl, C 1-4 Heteroalkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, 3-10 member heterocycloalkyl, C 6-10 Selected from aryls, 6-10 member heteroaryls, or R 20 and R 21 These may form a 3- to 10-membered heterocycloalkyl group together with the nitrogen atom to which they are bonded. n is 0, 1, 2, 3, or 4.

[0085] In some embodiments, ring A is C 3-6 Cycloalkyl, 3-10 member heterocycloalkyl, C 6-10 Selected from aryls and 6-10 membered heteroaryls. In some embodiments, ring A is C 3-6 Cycloalkyl, 3-10 member heterocycloalkyl, and C 6-10 Selected from aryl rings. In some embodiments, ring A is C 3-6 Selected from cycloalkyl and 3-10 membered heterocycloalkyl. In some embodiments, ring A is a 3-10 membered heterocycloalkyl. In some embodiments, ring A is C 6-10 It is an aryl compound. In some embodiments, ring A is a 6- to 10-membered heteroaryl compound. In some embodiments, ring A is C 3-6 It is a cycloalkyl. In some embodiments, ring A is

[0086] [ka] It is selected from the group consisting of the following.

[0087] In some embodiments, ring A is

[0088] [ka] In some embodiments, ring A is

[0089] [ka] In some embodiments, ring A is

[0090] [ka] In some embodiments, ring A is

[0091] [ka] In some embodiments, ring A is

[0092] [ka] In some embodiments, ring A is

[0093] [ka] In some embodiments, ring A is

[0094] [ka] In some embodiments, ring A is

[0095] [ka] That is the case.

[0096] In some embodiments, ring A is

[0097] [ka] It is selected from the group consisting of the following.

[0098] In some embodiments, ring A is

[0099] [ka] It is selected from the group consisting of the following.

[0100] In some embodiments, ring A is

[0101] [ka] It is selected from the group consisting of the following.

[0102] In some embodiments, ring A is

[0103] [ka] In some embodiments, ring A is

[0104] [ka] In some embodiments, ring A is

[0105] [ka] In some embodiments, ring A is

[0106] [ka] In some embodiments, ring A is

[0107] [ka] In some embodiments, ring A is

[0108] [ka] In some embodiments, ring A is

[0109] [ka] In some embodiments, ring A is

[0110] [ka] In some embodiments, ring A is

[0111] [ka] In some embodiments, ring A is

[0112] [ka] In some embodiments, ring A is

[0113] [ka] In some embodiments, ring A is

[0114] [ka] In some embodiments, ring A is

[0115] [ka] In some embodiments, ring A is

[0116] [ka] In some embodiments, ring A is

[0117] [ka] In some embodiments, ring A is

[0118] [ka] In some embodiments, ring A is

[0119] [ka] In some embodiments, ring A is

[0120] [ka] In some embodiments, ring A is

[0121] [ka] In some embodiments, ring A is

[0122] [ka] In some embodiments, ring A is

[0123] [ka] In some embodiments, ring A is

[0124] [ka] In some embodiments, ring A is

[0125] [ka] In some embodiments, ring A is

[0126] [ka] In some embodiments, ring A is

[0127] [ka] In some embodiments, ring A is

[0128] [ka] In some embodiments, ring A is

[0129] [ka] In some embodiments, ring A is

[0130] [ka] In some embodiments, ring A is

[0131] [ka] In some embodiments, ring A is

[0132] [ka] In some embodiments, ring A is

[0133] [ka] In some embodiments, ring A is

[0134] [ka] That is the case.

[0135] In some embodiments, X 1 , X 2 , X 3 , and X 4 None of these are N. In some embodiments, X 1 , X 2 , X 3 , and X 4 One of them is N. In some embodiments, X 1 , X 2 , X 3 , and X 4 Two of these are N. In some embodiments, X 1 , X 2 , X 3 , and X 4 Three of these are N. In some embodiments, X 1 , X 2 , X 3 , and X 4 In some embodiments, X 1 In some embodiments, X 2 In some embodiments, X 3 In some embodiments, X 4 In some embodiments, X 1 and X 2 In some embodiments, X 1 and X 3 In some embodiments, X 1 and X 4 In some embodiments, X 2 and X 3 In some embodiments, X 2 and X 4 In some embodiments, X 3 and X 4In some embodiments, X 1 , X 2 , and X 3 In some embodiments, X 1 , X 2 , and X 4 In some embodiments, X 1 , X 3 , and X 4 In some embodiments, X 2 , X 3 , and X 4 In some embodiments, X 1 , X 2 , X 3 , and X 4 This is N.

[0136] In some embodiments, R 11 , R 12 , R 13 , and R 14 These are H, Halo, -CN, and -OR, respectively, independently. 18 , -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 , -C(O)R 18 , -C(O)OR 18 , -NR 19 C(O)R 18 , C1-6 alkyl, and C 1-6 Selected from haloalkyls. In some embodiments, R 11 , R 12 , R 13 , and R 14 These are H, Halo, -CN, and -OR, respectively, independently. 18 , -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 , -C(O)R 18 , -C(O)OR 18 , and -NR 19 C(O)R 18Selected from. In some embodiments, R 11 , R 12 , R 13 , and R 14 These are H, Halo, -CN, and -OR, respectively, independently. 18 , and -NR 16 R 17 Selected from. In some embodiments, R 11 , R 12 , R 13 , and R 14 One of them is a halo, and the others are H. In some embodiments, R 11 is chloroform, R 12 , R 13 , and R 14 These are H.

[0137] In some embodiments, R 1 H, C1-6 alkyl, C 1-6 Haloalkyl and C 3-6 Selected from cycloalkyl. In some embodiments, R 1 is a C1-6 alkyl group. In some embodiments, R 1 C 1-6 In some embodiments, R 1 C 3-6 It is cycloalkyl. In some embodiments, R 1 R is selected from H, Me, Et, n-Pr, i-Pr, -CF3, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. In some embodiments, R 1 is selected from H, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. In some embodiments, R 1 is selected from Me, Et, n-Pr, and i-Pr. In some embodiments, R 1 is Me. In some embodiments, R 1 H is H.

[0138] In some embodiments, R 2 is -CN, -OR 18-SOR 15 , -SO2R 15 , -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 , -C(O)R 18 , -C(O)OR 18 , -NR 19 C(O)R 18 , -NR 19 C(O)NR 16 R 17 , -NR 19 SO2R 15 , -NR 19 SO2NR 16 R 17 , C1-6 alkyl, C 1-6 Haloalkyl and C 3-6 Selected from cycloalkyl. In some embodiments, R 2 R is selected from halo, -CN, -OH, -OMe, -OEt, -NH2, -NHMe, -NMe2, Me, Et, n-Pr, i-Pr, -CF3, and cyclopropyl. In some embodiments, R 2 is selected from Me, Et, n-Pr, and i-Pr. In some embodiments, R 2 is Me. In some embodiments, R 2 is -CF3. In some embodiments, R 2 It is cyclopropyl.

[0139] In some embodiments, R 3 H, C1-6 alkyl, C 1-6 Haloalkyl, C 3-6 Selected from cycloalkyls and 3- to 10-membered heterocycloalkyls. In some embodiments, R 3 is a C1-6 alkyl group. In some embodiments, R 3 C 1-6 In some embodiments, R 3 C 3-6 It is cycloalkyl. In some embodiments, R 3is a 3- to 10-membered heterocycloalkyl group. In some embodiments, R 3 R is selected from H, Me, Et, -CF3, and cyclopropyl. In some embodiments, R 3 H is H. In some embodiments, R 3 is Me. In some embodiments, R 3 It is -CF3.

[0140] In some embodiments, R 2 is Me, and R 3 H is H.

[0141] In some embodiments, R 4 C 6-10 Aryl, 6-10 member heteroaryl, -O(C 0-4 Alkyl)C 3-6 Cycloalkyl, -O(C 0-4 Alkyl)(3-10 member heterocycloalkyl), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl) (6-10 member heteroaryl), -O(C 0-4 Alkyl)C(O)OR 18 , -O(C 0-4 Alkyl)C(O)NR 19 SO2R 15 , -O(C 0-4 Alkyl)SO2NR 19 C(O)R 18 , -O(C 3-6 Cycloalkyl)C 3-6 Cycloalkyl, -O(C 3-6 Cycloalkyl) (3-10 member heterocycloalkyl), -O(C 3-6 Cycloalkyl)C 6-10 Aryl, -O(C 3-6 Cycloalkyl) (6-10 member heteroaryl), -O(C 3-6 Cycloalkyl)C(O)OR 18 ,-(C 1-4 Alkyl)C 3-6 Cycloalkyl, -(C 1-4Alkyl) (3-10 member heterocycloalkyl), -(C 1-4 Alkyl)C 6-10 Ariel, -(C 1-4 Alkyl) (6-10 member heteroaryl), and -(C 1-4 Alkyl)C(O)OR 18 Selected from, each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently C 1-4 Alkyl, oxo, halo, -OR 18 -CN, -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 , -C(O)R 18 , -C(O)OR 18 ,-(C 1-4 Alkyl)OC(O)(C 1-4 Alkyl), -(C 1-4 Alkyl)OC(O)OR 18 , -NR 19 C(O)R 18 , -NR 19 C(O)NR 16 R 17 , -NR 19 SO2R 15 , and -NR 19 SO2NR 16 R 17 Optionally substituted with one or more substituents selected from, R 4’ and R 4” These are, independently, H, C1-6 alkyl, and C 1-6 Haloalkyl and C 3-6 Selected from cycloalkyls, where each alkyl and cycloalkyl is independently halo, -OR 18 -CN and -NR 16 R 17 Optionally substituted with one or more substituents selected from, or R 3 H is, R 4 , R 4’ , and R 4”Together with the carbon atoms to which they are bonded, -C(O)R 18 Alternatively, they form a 6-10 member heteroaryl group.

[0142] In some embodiments, R 4 These are 6-10 member heteroaryl compounds, -O(C 0-4 Alkyl)(3-10 member heterocycloalkyl), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl) (6-10 member heteroaryl), -O(C 0-4 Alkyl)C(O)OR 18 , -O(C 0-4 Alkyl)C(O)NR 19 SO2R 15 , -O(C 0-4 Alkyl)SO2NR 19 C(O)R 18 , -O(C 3-6 Cycloalkyl) (6-10 member heteroaryl), -O(C 3-6 Cycloalkyl)C(O)OR 18 ,-(C 1-4 Alkyl) (6-10 member heteroaryl), and -(C 1-4 Alkyl)C(O)OR 18 Selected from, where each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently C 1-4 Alkyl, oxo, halo, -OR 18 -CN, -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 , -C(O)OR 18 ,-(C 1-4 Alkyl)OC(O)(C 1-4 Alkyl), -(C 1-4 Alkyl)OC(O)OR 18 , -NR 19 C(O)R 18 , -NR 19 C(O)NR 16 R 17, -NR 19 SO2R 15 , and -NR 19 SO2NR 16 R 17 Optionally substituted with one or more substituents selected from, R 4’ and R 4” Both are H, or R 3 H is, R 4 , R 4’ , and R 4” Together with the carbon atoms to which they are bonded, -C(O)R 18 Alternatively, they form a 6-10 member heteroaryl group.

[0143] In some embodiments, R 4 These are 6-10 member heteroaryl compounds, -O(C 0-4 Alkyl)(3-10 member heterocycloalkyl), -O(C 0-4 Alkyl)C 6-10 Aryl, -O(C 0-4 Alkyl) (6-10 member heteroaryl), -O(C 0-4 Alkyl)C(O)OR 18 , -O(C 0-4 Alkyl)C(O)NR 19 SO2R 15 , -O(C 0-4 Alkyl)SO2NR 19 C(O)R 18 , -O(C 3-6 Cycloalkyl) (6-10 member heteroaryl), -O(C 3-6 Cycloalkyl)C(O)OR 18 ,-(C 1-4 Alkyl) (6-10 member heteroaryl), and -(C 1-4 Alkyl)C(O)OR 18 Selected from, where each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently C 1-4 Alkyl, oxo, halo, -OR 18 -CN, -NR 16 R17 -C(O)NR 16 R 17 -SO2NR 16 R 17 , -C(O)OR 18 ,-(C 1-4 Alkyl)OC(O)(C 1-4 Alkyl), -(C 1-4 Alkyl)OC(O)OR 18 , -NR 19 C(O)R 18 , -NR 19 C(O)NR 16 R 17 , -NR 19 SO2R 15 , and -NR 19 SO2NR 16 R 17 Optionally substituted with one or more substituents selected from, R 4’ and R 4” Both are H.

[0144] In some embodiments, R 3 H is, R 4 , R 4’ , and R 4” Together with the carbon atoms to which they are bonded, -C(O)R 18 Alternatively, they form a 6-10 member heteroaryl group.

[0145] In some embodiments, R 3 H is, R 4 , R 4’ , and R 4” Together with the carbon atoms to which they are bonded, -C(O)R 18 Alternatively, they form a 6-10 member heteroaryl group.

[0146] In some embodiments, R 3 H is, R 4 , R 4’ , and R4” These, together with the carbon atoms to which they are bonded, form 6- to 10-membered heteroaryl groups.

[0147] In some embodiments, R 5 H, C1-6 alkyl, C 1-6 Haloalkyl and C 3-6 Selected from cycloalkyl. In some embodiments, R 5 C 1-6 In some embodiments, R 5 C 3-6 It is cycloalkyl. In some embodiments, R 5 is a C1-6 alkyl group. In some embodiments, R 5 is Me, Et, n-Pr, or i-Pr. In some embodiments, R 5 is Me. In some embodiments, R 5 H is H.

[0148] In some embodiments, R 6 and R 7 These are H and -(C) respectively, independently. 1-4 Alkyl)C 3-6 Cycloalkyl, -(C 1-4 Alkyl) (3-10 member heterocycloalkyl), -(C 1-4 Alkyl)C 6-10 Aryl, and -(C 1-4 Selected from alkyl (6-10 member heteroaryl), where each alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is independently oxo, halo, -OR 18 , C1-4 alkyl, C1-4 alkoxy, C 1-4 Heteroalkyl, C 1-4 Haloalkyl, -CN, and -NR 20 R 21 Optionally substituted with one or more substituents selected from, or R 6 and R 7These, together with the nitrogen atom to which they are bonded, form oxo, halo, -OR 18 , C1-4 alkyl, C1-4 alkoxy, C 1-4 Heteroalkyl, C 1-4 Haloalkyl, -CN, and -NR 20 R 21 It forms a 3- to 10-membered heterocycloalkyl group, which is optionally substituted with one or more substituents selected from the following.

[0149] In some embodiments, R 6 and R 7 These are H and -(C) respectively, independently. 1-4 Selected from alkyl (3-10 member heterocycloalkyl), where each alkyl and heterocycloalkyl is independently halo, -OR 18 -CN and -NR 20 R 21 It is optionally substituted with one or more substituents selected from the following.

[0150] In some embodiments, R 6 and R 7 One of them is H, and the other is

[0151] [ka] In some embodiments, R 6 H is R 7 teeth

[0152] [ka] In some embodiments, R 7 H is R 6 teeth

[0153] [ka] That is the case.

[0154] In some embodiments, R 8 and R 9 These are H, Halo, -CN, and -OR, respectively, independently. 18 , -SO2R 15 , -NR 16 R 17 -C(O)NR 16 R 17 -SO2NR 16 R 17 , -C(O)OR 18 , -NR 19 C(O)R 18 , -NR 19 SO2R 15 Selected from. In some embodiments, R 8 and R 9 These are H, Halo, -CN, and -OR, respectively, independently. 18 , and -NR 16 R 17 Selected from. In some embodiments, R 8 and R 9 Both are H.

[0155] In some embodiments, n is 0, 1, 2, 3, or 4. In some embodiments, n is 0, 1, 2, or 3. In some embodiments, n is 0, 1, or 2. In some embodiments, n is 0 or 1. In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.

[0156] In some embodiments, the compound of formula (I) is formula (IA):

[0157] [ka] It is represented by [this].

[0158] In some embodiments, the compound of formula (I) is formula (IB):

[0159] [ka] It is represented by [this].

[0160] In some embodiments, the compound of formula (I) is formula (IC), formula (ID), formula (IE), or formula (IF):

[0161] [ka] It is represented by [this].

[0162] In some embodiments, the compound of formula (I) is of formula (IC):

[0163] [ka] It is represented by [this].

[0164] In some embodiments, the compound of formula (I) is formula (ID):

[0165] [ka] It is represented by [this].

[0166] In some embodiments, the compound of formula (I) is formula (IE):

[0167] [ka] It is represented by [this].

[0168] In some embodiments, the compound of formula (I) is formula (IF):

[0169] [ka] It is represented by [this].

[0170] In some embodiments, the compound is

[0171] [ka]

[0172] [ka]

[0173] [ka]

[0174] [ka]

[0175] [ka]

[0176] [ka]

[0177] [ka]

[0178] [ka]

[0179] [ka]

[0180] [ka]

[0181] [ka] It is selected from the group consisting of the following.

[0182] In some embodiments, the compound is

[0183] [ka] In some embodiments, the compound is

[0184] [ka] In some embodiments, the compound is

[0185] [ka] In some embodiments, the compound is

[0186] [ka] In some embodiments, the compound is

[0187] [ka] In some embodiments, the compound is

[0188] [ka] In some embodiments, the compound is

[0189] [ka] In some embodiments, the compound is

[0190] [ka] In some embodiments, the compound is

[0191] [ka] In some embodiments, the compound is

[0192] [ka] In some embodiments, the compound is

[0193] [ka] In some embodiments, the compound is

[0194] [ka] In some embodiments, the compound is

[0195] [ka] In some embodiments, the compound is

[0196] [ka] In some embodiments, the compound is

[0197] [ka] In some embodiments, the compound is

[0198] [ka] In some embodiments, the compound is

[0199] [ka] In some embodiments, the compound is

[0200] [ka] In some embodiments, the compound is

[0201] [ka] In some embodiments, the compound is

[0202] [ka] In some embodiments, the compound is

[0203] [ka] In some embodiments, the compound is

[0204] [ka] In some embodiments, the compound is

[0205] [ka] In some embodiments, the compound is

[0206] [ka] In some embodiments, the compound is

[0207] [ka] In some embodiments, the compound is

[0208] [ka] In some embodiments, the compound is

[0209] [ka] In some embodiments, the compound is

[0210] [ka] In some embodiments, the compound is

[0211] [ka] In some embodiments, the compound is

[0212] [ka] In some embodiments, the compound is

[0213] [ka] In some embodiments, the compound is

[0214] [ka] In some embodiments, the compound is

[0215] [ka] In some embodiments, the compound is

[0216] [ka] In some embodiments, the compound is

[0217] [ka] In some embodiments, the compound is

[0218] [ka] In some embodiments, the compound is

[0219] [ka] In some embodiments, the compound is

[0220] [ka] In some embodiments, the compound is

[0221] [ka] In some embodiments, the compound is

[0222] [ka] In some embodiments, the compound is

[0223] [ka] In some embodiments, the compound is

[0224] [ka] In some embodiments, the compound is

[0225] [ka] In some embodiments, the compound is

[0226] [ka] In some embodiments, the compound is

[0227] [ka] In some embodiments, the compound is

[0228] [ka] In some embodiments, the compound is

[0229] [ka] In some embodiments, the compound is

[0230] [ka] In some embodiments, the compound is

[0231] [ka] In some embodiments, the compound is

[0232] [ka] In some embodiments, the compound is

[0233] [ka] In some embodiments, the compound is

[0234] [ka] In some embodiments, the compound is

[0235] [ka] In some embodiments, the compound is

[0236] [ka] In some embodiments, the compound is

[0237] [ka] In some embodiments, the compound is

[0238] [ka] In some embodiments, the compound is

[0239] [ka] In some embodiments, the compound is

[0240] [ka] In some embodiments, the compound is

[0241] [ka] In some embodiments, the compound is

[0242] [ka] In some embodiments, the compound is

[0243] [ka] In some embodiments, the compound is

[0244] [ka] In some embodiments, the compound is

[0245] [ka] In some embodiments, the compound is

[0246] [ka] In some embodiments, the compound is

[0247] [ka] In some embodiments, the compound is

[0248] [ka] In some embodiments, the compound is

[0249] [ka] In some embodiments, the compound is

[0250] [ka] In some embodiments, the compound is

[0251] [ka] In some embodiments, the compound is

[0252] [ka] In some embodiments, the compound is

[0253] [ka] In some embodiments, the compound is

[0254] [ka] In some embodiments, the compound is

[0255] [ka] In some embodiments, the compound is

[0256] [ka] In some embodiments, the compound is

[0257] [ka] In some embodiments, the compound is

[0258] [ka] In some embodiments, the compound is

[0259] [ka] In some embodiments, the compound is

[0260] [ka] In some embodiments, the compound is

[0261] [ka] In some embodiments, the compound is

[0262] [ka] That is the case.

[0263] In another embodiment, a pharmaceutical composition comprising a compound described herein or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient is provided herein.

[0264] In another embodiment, a method for treating a disease or disorder in a patient in need thereof is provided herein, comprising administering a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, to a patient in need thereof.

[0265] In some embodiments, the disease or disorder is cancer. In some embodiments, the cancer is selected from leukemia, breast cancer, prostate cancer, ovarian cancer, colon cancer, cervical cancer, lung cancer, lymphoma, and liver cancer. In some embodiments, the cancer is leukemia. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is colon cancer. In some embodiments, the cancer is cervical cancer. In some embodiments, the cancer is lung cancer. In some embodiments, the cancer is lymphoma. In some embodiments, the cancer is liver cancer.

[0266] How to use In one embodiment, the present invention provides a method for treating a proliferative disorder in a subject requiring such treatment, comprising administering a compound having formula (I), as further disclosed herein, to the subject. In some embodiments, the method for treating the proliferative disorder comprises administering a CDK9 inhibitor to the subject. In some embodiments, the compound having formula (I) is a CDK9 inhibitor. In some embodiments, the proliferative disorder is a cancerous condition. In some further embodiments, the cancerous condition is a cancer selected from the group consisting of leukemia, breast cancer, prostate cancer, ovarian cancer, colon cancer, cervical cancer, lung cancer, lymphoma, and liver cancer. In some embodiments, the cancerous condition is liver cancer.

[0267] In some embodiments, the CDK9 inhibitors disclosed herein are highly targeted to the liver. In some embodiments, the CDK9 inhibitors disclosed herein exhibit superior liver targeting compared to known CDK9 inhibitors. In some embodiments, the CDK9 inhibitors disclosed herein accumulate in the liver while avoiding peripheral exposure to nearby tissues. In some embodiments, the CDK9 inhibitors disclosed herein exhibit reduced peripheral exposure to nearby tissues compared to known CDK9 inhibitors. In some embodiments, the CDK9 inhibitors disclosed herein have reduced toxicity compared to known CDK9 inhibitors.

[0268] In further embodiments, the present invention provides a method for treating a cancerous condition in which a compound having formula (I) (e.g., a CDK9 inhibitor) is effective in one or more methods of inhibiting the proliferation of cancer cells, inhibiting the metastasis of cancer cells, reducing the severity or incidence of symptoms associated with the presence of cancer cells, and promoting an immune response against tumor cells. In some embodiments, the method comprises administering a therapeutically effective amount of a compound having formula (I) to cancer cells. In some embodiments, the compound having formula (I) is a CDK9 inhibitor. In some embodiments, the administration is performed in vitro. In other embodiments, the administration is performed in vivo.

[0269] As used herein, a therapeutically effective dose of a CDK9 inhibitor means an amount sufficient to accomplish an intended use, including but not limited to the treatment of a disease, as defined herein. The use of sub-therapeutic doses of a CDK9 inhibitor to treat an intended disease condition is also contemplated in the methods described herein.

[0270] The dosage of CDK9 inhibitors may vary depending on the intended use (in vitro or in vivo), or the subject and disease state being treated, such as the subject's weight and age, the severity of the disease state, the mode of administration, and similar factors, which can be easily determined by those skilled in the art.

[0271] Measuring the inhibition of the biological effects of CDK9 may involve performing assays on biological samples, such as samples derived from the subject. Depending on the assay, one of a variety of samples may be selected. Examples of samples, but not limited to, include blood samples (e.g., plasma or serum), exhaled condensate samples, bronchoalveolar lavage fluid, sputum samples, urine samples, and tissue samples.

[0272] Subjects being treated with CDK9 inhibitors may be monitored to determine the effectiveness of the treatment, and the treatment regimen may be adjusted based on the subject's physiological response to the treatment. For example, if the inhibition of the biological effect of CDK9 degradation is above or below a threshold, the dose or frequency of administration may be reduced or increased, respectively. The method may further include continuing treatment if it is determined that the treatment is effective. The method may include maintaining, tapering, reducing, or discontinuing the dose of the compound in treatment if it is determined that the treatment is effective. The method may include increasing the dose of the compound in treatment if it is determined that the treatment is not effective. Alternatively, the method may include discontinuing treatment if it is determined that the treatment is not effective. In some embodiments, treatment with a CDK9 inhibitor is discontinued if the inhibition of the biological effect is above or below a threshold, for example, if there is no response or an adverse reaction. The biological effect may be a change in any of a variety of physiological indicators.

[0273] Generally, CDK9 inhibitors are compounds that inhibit one or more biological effects of CDK9. Such biological effects may be inhibited by approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more.

[0274] In several other embodiments, the method of the subject is useful for treating disease conditions associated with CDK9. Any disease condition directly or indirectly resulting from abnormal activity or expression levels of CDK9 may be the intended disease condition. In some embodiments, the disease condition is a proliferative disorder, including but not limited to cancer, such as those described herein. In some embodiments, the disease condition is cancer. The role of CDK9 in tumorigenesis and tumor progression has been linked to many human cancers. Consequently, agents targeting CDK9 have therapeutic value.

[0275] The data presented in the examples described later in this specification demonstrate the anticancer effects of CDK9 inhibitors. Therefore, the method described in this subject is particularly useful for treating proliferative disorders, such as neoplasms.

[0276] In some embodiments, the methods of administering the CDK9 inhibitors described herein are applied to the treatment of cancers of the blood, breast, prostate, ovaries, colon, cervix, lung, lymph nodes, liver, or any combination thereof.

[0277] Treatment effectiveness In some embodiments, therapeutic efficacy is measured based on the effect of treating proliferative disorders such as cancer. Generally, with respect to the treatment of proliferative disorders (e.g., cancer, regardless of whether it is benign or malignant), the therapeutic efficacy of the methods and compositions of the present invention may be measured by the degree to which the methods and compositions promote inhibition of tumor cell proliferation, inhibition of tumor angiogenesis, eradication of tumor cells, reduction of tumor growth rate, and / or reduction of the size of at least one tumor. Several parameters to be considered in determining therapeutic efficacy are discussed herein. The appropriate combination of parameters for a particular situation may be established by the clinician. The progress of the methods of the present invention in the treatment of cancer (e.g., reduction of tumor size or eradication of cancer cells) can be confirmed using any preferred method, such as methods currently used clinically to track tumor size or cancer progression. The primary efficacy parameter used to evaluate the treatment of cancer with the methods and compositions of the present invention is preferably reduction of tumor size. The size of the tumor can be determined by using any suitable technique, such as dimensional measurement, or by estimating the tumor volume using available computer software, such as the FreeFlight software developed at Wake Forest University, which allows for accurate estimation of tumor volume. The size of the tumor can also be determined by visualizing the tumor using, for example, CT, ultrasound, SPECT, spiral CT, MRI, photography, and similar methods. In embodiments where the tumor is surgically removed after the completion of the treatment period, the presence and size of the tumor tissue can be determined by macroscopic analysis of the tissue to be removed and / or pathological analysis of the removed tissue.

[0278] In some preferred embodiments, tumor growth is stabilized as a result of the present invention's methods and compositions (i.e., one or more tumors do not increase in size by more than 1%, 5%, 10%, 15%, or 20%, and / or metastasize). In some embodiments, the tumors are stabilized for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or more weeks. In some embodiments, the tumors are stabilized for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or more months. In some embodiments, the tumors are stabilized for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more years. Preferably, the present invention's methods reduce tumor size by at least about 5% (e.g., at least about 10%, 15%, 20%, or 25%). More preferably, the size of the tumor is reduced by at least about 30% (e.g., at least about 35%, 40%, 45%, 50%, 55%, 60%, or 65%). Even more preferably, the size of the tumor is reduced by at least about 70% (e.g., at least about 75%, 80%, 85%, 90%, or 95%). Most preferably, the tumor is completely removed or reduced to below the detection level. In some embodiments, the subject remains tumor-free (e.g., in remission) for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or more weeks after treatment. In some embodiments, the subject remains tumor-free for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or more months after treatment. In some embodiments, the subjects remain tumor-free for at least approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more years after treatment.

[0279] In some embodiments, the effectiveness of the present invention in reducing tumor size can be determined by measuring the percentage of necrotic (i.e., dead) tissue of the tumor surgically removed after the end of the treatment period. In some further embodiments, the treatment is therapeutically effective when the necrotic rate of the removed tissue is greater than about 20% (e.g., at least about 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%), more preferably about 90% or greater (e.g., about 90%, 95%, or 100%). Most preferably, the necrotic rate of the removed tissue is 100%, i.e., tumor tissue is absent or undetectable.

[0280] The effectiveness of the present invention's methods can be determined by a number of secondary parameters. Examples of secondary parameters, but not limited to, include, detection of new tumors, detection of tumor antigens or markers (e.g., CEA, PSA, or CA-125), biopsy, surgical downstaging (i.e., surgical stage conversion from unresectable to resectable), PET scan, survival time, progression-free survival, time to disease progression, quality of life assessments such as Clinical Benefit Response Assessment, and similar factors, all of which can indicate the overall progression (or regression) of cancer in humans. Biopsy is particularly effective in detecting the eradication of cancer cells within tissue. Radioimmunodetection (RAID) is used to locate and stage tumors using serum levels of tumor-produced and / or tumor-associated markers (antigens) ("tumor markers" or "tumor-associated antigens"), and can be useful as a pre-treatment diagnostic predictor, a post-treatment recurrence diagnostic indicator, and an indicator of post-treatment treatment effectiveness. Examples of tumor markers or tumor-associated antigens that can be evaluated as indicators of therapeutic efficacy include, but are not limited to, carcinoembryonic antigen (CEA), prostate-specific antigen (PSA), CA-125, CA19-9, ganglioside molecules (e.g., GM2, GD2, and GD3), MART-1, heat shock proteins (e.g., gp96), sialyl Tn (STn), tyrosinase, MUC-1, HER-2 / neu, c-erb-B2, KSA, PSMA, p53, RAS, EGF-R, VEGF, MAGE, and gp100. Other tumor-associated antigens are known in the art. Furthermore, RAID technology can be combined with endoscopic detection systems to efficiently differentiate small tumors from surrounding tissue (see, for example, U.S. Patent No. 4,932,412).

[0281] In an additional preferred embodiment, treatment of cancer in a human patient according to the present invention is demonstrated by one or more of the following results: (a) complete disappearance of the tumor (i.e., complete response); (b) a reduction in tumor size of about 25% to about 50% compared to the tumor size before treatment, at least 4 weeks after the end of the treatment period; (c) a reduction in tumor size of at least about 50% compared to the tumor size before treatment, at least 4 weeks after the end of the treatment period; and (d) a reduction of at least about 2% (e.g., about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%) in a specific tumor-associated antigen level about 4 to 12 weeks after the end of the treatment period, compared to the tumor-associated antigen level before treatment. A reduction of at least 2% in tumor-associated antigen levels is preferred, but any reduction in tumor-associated antigen levels is evidence of treatment of cancer in a patient by the present invention. For example, in the case of unresectable locally advanced pancreatic cancer, treatment can be demonstrated by a decrease of at least 10% in CA19-9 tumor-associated antigen levels 4 to 12 weeks after the end of the treatment period compared to the CA19-9 level before the treatment period. Similarly, in the case of locally advanced rectal cancer, treatment can be demonstrated by a decrease of at least 10% in CEA tumor-associated antigen levels 4 to 12 weeks after the end of the treatment period compared to the CEA level before the treatment period.

[0282] With regard to quality of life assessments such as Clinical Benefit Response Criteria, the therapeutic benefits of treatment according to the present invention can be demonstrated in terms of pain intensity, analgesic consumption, and / or Karnofsky Performance Scale score. Alternatively, in addition, cancer treatment in human patients is demonstrated by (a) a reduction of at least 50% (e.g., at least 60%, 70%, 80%, 90%, or 100%) in the patient's reported pain intensity during any four consecutive weeks within 12 weeks after the end of treatment compared to the pain intensity reported by the patient before treatment; (b) a reduction of at least 50% (e.g., at least 60%, 70%, 80%, 90%, or 100%) in the patient's reported analgesic consumption during any four consecutive weeks within 12 weeks after the end of treatment compared to the analgesic consumption reported by the patient before treatment; and / or (c) an increase of at least 20 points (e.g., at least 30, 50, 70, or 90 points) in the patient's reported Karnofsky Performance Scale score during any four consecutive weeks within 12 weeks after the end of the treatment period compared to the Karnofsky Performance Scale score reported by the patient before the treatment period.

[0283] Results from alternative or additional reference trials and / or other trials may also demonstrate the efficacy of a treatment, but the treatment of proliferative disorders (e.g., cancers, regardless of whether they are benign or malignant) in human patients is preferably demonstrated by one or more (any combination) of the aforementioned results.

[0284] In some embodiments, tumor size is preferably reduced as a result of the present invention without significant adverse events in the subject. Adverse events are classified or “graded” by the National Cancer Institute (NCI) Cancer Therapy Evaluation Program (CTEP), where Grade 0 represents minimal adverse side effects and Grade 4 represents the most severe adverse events. Preferably, the present invention is associated with minimal adverse events graded by CTEP / NCI, e.g., Grade 0, Grade 1, or Grade 2 adverse events. However, as discussed herein, while reduction in tumor size is desirable, it is not essential because the actual size of the tumor may not decrease despite the eradication of tumor cells. Eradication of cancer cells is sufficient to recognize a therapeutic effect. Similarly, any reduction in tumor size is sufficient to recognize a therapeutic effect.

[0285] Further information on the detection, monitoring, and classification of various cancers in humans can be found in Cancer Facts and Figures 2001, American Cancer Society, New York, NY, and International Patent Application WO01 / 24684. Therefore, clinicians can use standard tests to determine the effectiveness of various embodiments of the inventive method in the treatment of cancer. However, in addition to tumor size and spread, clinicians may also consider the patient's quality of life and survival when evaluating the effectiveness of treatment.

[0286] In some embodiments, administration of a CDK9 inhibitor provides improved therapeutic efficacy. Improved efficacy may be measured using any method known in the art (including, but not limited to, the methods described herein). In some embodiments, improved therapeutic efficacy is an improvement of at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 95%, 100%, 110%, 120%, 150%, 200%, 300%, 400%, 500%, 600%, 700%, 1000%, or more, using an appropriate measure (e.g., reduction in tumor size, duration of tumor size stability, duration without metastatic events, disease-free survival). Improved efficacy may also be expressed as a multiplier improvement of at least approximately 2x, 3x, 4x, 5x, 6x, 7x, 8x, 9x, 10x, 20x, 30x, 40x, 50x, 60x, 70x, 80x, 90x, 100x, 1000x, 10000x, or more, using an appropriate measure (e.g., reduction in tumor size, duration of stable tumor size, duration without metastatic events, disease-free survival).

[0287] Pharmaceutical composition The compositions of this disclosure may be formulated into any suitable pharmaceutical formulation. The pharmaceutical compositions of this disclosure typically contain an active ingredient (e.g., a compound of formula (I), or a pharmaceutically acceptable salt and / or coordination complex thereof) and one or more pharmaceutically acceptable excipients or carriers (including, but not limited to, inert solid diluents and fillers, diluents, sterile aqueous solutions and various organic solvents, permeation enhancers, solubilizers, and adjuvants). The compositions of this disclosure may be formulated into any suitable pharmaceutical formulation. In some embodiments, the pharmaceutically acceptable carrier or excipient is selected from water, alcohol, glycerol, chitosan, alginate, chondroitin, vitamin E, mineral oil, and dimethyl sulfoxide (DMSO).

[0288] Pharmaceutical formulations may be provided in any preferred form, which may depend on the route of administration. In some embodiments, the pharmaceutical compositions disclosed herein can be formulated into dosage forms for administration to a subject. In some embodiments, pharmaceutical compositions are formulated for oral administration, intravenous administration, intra-arterial administration, aerosol administration, parenteral administration, oral administration, topical administration, transdermal administration, rectal administration, intramuscular administration, subcutaneous administration, intraosseous administration, intranasal administration, intrapulmonary administration, transmucosal administration, inhalation administration, and / or intraperitoneal administration. In some embodiments, the dosage form is formulated for oral administration. For example, pharmaceutical compositions can be formulated in the form of pills, tablets, capsules, inhalation formulations, liquid suspensions, liquid emulsions, gels, or powders. In some embodiments, pharmaceutical compositions can be formulated as unit doses in the form of liquids, gels, semi-liquids, semi-solids, or solids.

[0289] The dosage of each compound will depend on the mammal being treated, the severity of the disorder or condition, the rate of administration, the biodistribution of the compound, and the discretion of the prescribing physician. However, the effective dose may range from approximately 0.001 to approximately 100 mg per kg of body weight per day, either as a single dose or in divided doses. In some cases, a dose level below the lower limit of the aforementioned range may be more than sufficient, while in other cases, a larger dose may be employed without causing any adverse side effects, for example, by dividing such a large dose into several smaller doses to be administered throughout the day. In some embodiments, the effective dose may be provided by pulse administration (i.e., administering the compound for several consecutive days, followed by several consecutive days of rest).

[0290] In some embodiments, the composition is provided in one or more unit doses. For example, the composition may be administered in doses of 1, 2, 3, 4, 5, 6, 7, 14, 30, 60, or more. Such amounts may be administered daily, for example, once, twice, or three times a day, or more. However, the fact that a dose is referred to herein as a daily dose should not be construed as meaning that the daily dose must be administered daily. For example, if one of the drugs is provided in a preferred sustained-release form, two or more daily doses may be administered at a lower frequency, for example, once every two days to once a month or longer, as a depot injection or oral prodrug. Most typically and conveniently for the target, a CDK9 inhibitor may be administered once daily, for example, in the morning, evening, or during the day.

[0291] The unit dose can be administered simultaneously or sequentially. This composition can be administered for a long treatment period. Exemplarily, the treatment period may be at least about one month, for example, at least about three months, at least about six months, or at least about one year. In some cases, administration can be continued substantially for the remainder of the subject's life.

[0292] In some embodiments, a CDK9 inhibitor may be administered as part of a treatment regimen that includes administering one or more second agents (e.g., 1, 2, 3, 4, 5, or more second agents) concurrently or sequentially with the CDK9 inhibitor. When administered sequentially, the CDK9 inhibitor may be administered before or after the one or more second agents. When administered concurrently, the CDK9 inhibitor and one or more second agents may be administered via the same route (e.g., injection at the same site, tablets taken orally at the same time), via different routes (e.g., tablets taken orally while being administered intravenously), or as part of the same combination (e.g., a solution containing the CDK9 inhibitor and one or more second agents).

[0293] Combination therapy according to the present invention may be effective over a wide range of dosages. For example, for the treatment of adults, possible dosages include 0.01–1000 mg, 0.5–100 mg, 1–50 mg per day, and 5–40 mg per day. The exact dosage will depend on the selected drug, route of administration, the form in which the compound is administered, the patient being treated, the patient's weight, and the physician's preference and experience.

[0294] Pharmaceutical compositions for oral administration In some embodiments, the Disclosure provides a pharmaceutical composition for oral administration comprising at least one compound of the Disclosure and a pharmaceutical excipient suitable for oral administration. The composition may be in the form of a solid, liquid, gel, semi-liquid, or semi-solid. In some embodiments, the composition further comprises a second drug.

[0295] In some embodiments, the present invention provides a solid pharmaceutical composition for oral administration comprising (i) a CDK9 inhibitor and (ii) a pharmaceutical excipient suitable for oral administration. In some embodiments, the composition further comprises (iii) a third or even a fourth agent. In some embodiments, each compound or agent is present in a therapeutically effective amount. In other embodiments, one or more compounds or agents are present in amounts less than or equal to a therapeutic amount, and the compounds or agents act synergistically to provide a therapeutically effective pharmaceutical composition.

[0296] Pharmaceutical compositions of the present disclosure suitable for oral administration may be presented as individual dosage forms, such as hard capsules or soft capsules, cachets, lozenges, tablets, liquids or aerosol sprays, dispersible powders or granules, syrups, or elixirs, each containing a predetermined amount of the active ingredient in powder or granules, solution, or suspension in an aqueous or non-aqueous liquid, oil-in-water emulsion, or water-in-oil liquid emulsion. Such dosage forms may be prepared by any pharmaceutical method, which typically involves the step of binding the active ingredient to a carrier. Generally, compositions are prepared by uniformly and tightly mixing the active ingredient with a liquid carrier or a finely divided solid carrier, or both, and then, if necessary, shaping the product into a desired form. For example, tablets may be prepared by compression or molding, with one or more adjuncts as optional. Compressed tablets can be prepared by mixing an active ingredient in a free-flowing form, such as powder or granules, with optionally excipients (e.g., binders, lubricants, inert diluents, and / or surfactants or dispersants), and compressing the mixture in a suitable machine. Molded tablets can be prepared by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0297] Because water can accelerate the decomposition of some compounds, this disclosure further encompasses anhydrous pharmaceutical compositions and dosage forms containing active ingredients. For example, in the pharmaceutical technology field, water (e.g., 5%) may be added as a means of simulating long-term storage to determine properties such as shelf life or the stability of a formulation over time. The anhydrous pharmaceutical compositions and dosage forms of this disclosure can be prepared by using anhydrous or low-moisture-containing ingredients and low-moisture or low-humidity conditions. Pharmaceutical compositions and dosage forms of this disclosure containing lactose can be anhydrous if substantial contact with moisture and / or humidity is expected during manufacturing, packaging, and / or storage. Anhydrous pharmaceutical compositions can be prepared and stored so as to maintain their anhydrous nature. Therefore, anhydrous compositions can be packaged using materials known to prevent exposure to water so that they can be included in suitable formulation kits. Examples of suitable packaging, but not limited to, include hermetically sealed foil, plastic or similar, unit dose containers, blister packs, and strip packs.

[0298] The active ingredient can be tightly mixed and combined with a pharmaceutical carrier according to conventional pharmaceutical formulation techniques. The carrier can take a wide variety of forms depending on the desired formulation for administration. When preparing compositions for oral dosage forms, for example, in the case of oral liquid formulations (e.g., suspensions, solutions, and elixirs) or aerosol formulations, any of the usual pharmaceutical media such as water, glycol, oil, alcohol, flavoring agents, preservatives, colorants, and the like can be used as the carrier. Alternatively, in the case of oral solid formulations, in some embodiments that do not employ lactose, carriers such as starch, sugars, microcrystalline cellulose, diluents, granulators, lubricants, binders, and disintegrants can be used. For example, suitable carriers for solid oral formulations include powders, capsules, and tablets. If desired, tablets can be coated using standard aqueous or non-aqueous techniques.

[0299] Suitable binders for use in pharmaceutical compositions and dosage forms include, but are not limited to, corn starch, potato starch or other starches, gelatin, natural and synthetic gums such as acacia, sodium alginate, alginic acid, other alginates, tragacanth powder, guar gum, cellulose and its derivatives (e.g., ethylcellulose, cellulose acetate, calcium carboxymethylcellulose, sodium carboxymethylcellulose), polyvinylpyrrolidone, methylcellulose, pregelatinized starch, hydroxypropyl methylcellulose, microcrystalline cellulose, and mixtures thereof.

[0300] Examples of fillers suitable for use in the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextrate, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, and mixtures thereof.

[0301] To provide tablets that disintegrate when exposed to an aqueous environment, disintegrants may be used in the compositions of this disclosure. Too much disintegrant may result in tablets that disintegrate in the bottle. Too little disintegrant may be insufficient for disintegration to occur, and the rate and extent of release of the active ingredient from the dosage form may be altered. To form the dosage forms of the compounds disclosed herein, a sufficient amount of disintegrant may be used, not too little or too much, so as not to alter the release of the active ingredient in an adverse manner. The amount of disintegrant used may vary depending on the type of formulation and mode of administration, which will be easily determined by those skilled in the art. A pharmaceutical composition may contain about 0.5 to about 15 weight percent of disintegrant or about 1 to about 5 weight percent of disintegrant. Disintegrants that can be used to form the pharmaceutical compositions and dosage forms of the present disclosure include, but are not limited to, agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polaritrin potassium, sodium starch glycolate, potato or tapioca starch, other starches, pregelatinized starch, other starches, clay, other algins, other celluloses, gums, or mixtures thereof.

[0302] Lubricants that can be used to form the pharmaceutical compositions and dosage forms of this disclosure include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oils (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, or mixtures thereof. Additional lubricants include, for example, thyroid silica gel, synthetic silica coagulation aerosols, or mixtures thereof. Lubricants may be optionally added in amounts less than about 1 weight percent of the pharmaceutical composition.

[0303] If an aqueous suspension and / or elixir is desired for oral administration, the active ingredient therein may be combined with various sweeteners or flavorings, colorants or dyes, and, if so desired, emulsifiers and / or suspending agents, along with diluents such as water, ethanol, propylene glycol, glycerin, and various combinations thereof.

[0304] Tablets may be uncoated or coated by known techniques to delay digestion and absorption in the gastrointestinal tract, thereby providing a sustained effect over a longer period. For example, time-delaying materials such as glyceryl monostearate or glyceryl distearate can be employed. Formulations for oral use may also be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with a water or oil medium, such as peanut oil, liquid paraffin, or olive oil.

[0305] Surfactants that can be used to form the pharmaceutical compositions and dosage forms of this disclosure include, but are not limited to, hydrophilic surfactants, lipophilic surfactants, and mixtures thereof. That is, a mixture of hydrophilic surfactants may be used, a mixture of lipophilic surfactants may be used, or a mixture of at least one hydrophilic surfactant and at least one lipophilic surfactant may be used.

[0306] Suitable hydrophilic surfactants typically have an HLB value of at least 10, while suitable lipophilic surfactants typically have an HLB value of about 10 or less. The empirical parameter used to characterize the relative hydrophilicity and hydrophobicity of nonionic amphiphilic compounds is the hydrophilic-lipophilic balance ("HLB" value). Surfactants with lower HLB values ​​are more lipophilic or hydrophobic and have greater solubility in oil, while surfactants with higher HLB values ​​are more hydrophilic and have greater solubility in aqueous solutions. Hydrophilic surfactants are generally considered to be compounds with an HLB value greater than about 10, as well as anionic, cationic, or zwitterionic compounds to which the HLB scale is generally not applicable. Similarly, lipophilic (i.e., hydrophobic) surfactants are compounds with an HLB value of about 10 or less. However, the HLB value of a surfactant is only a rough guideline commonly used to enable the formulation of industrial, pharmaceutical, and cosmetic emulsions.

[0307] Hydrophilic surfactants can be either ionic or nonionic. Suitable ionic surfactants include, but are not limited to, alkylammonium salts; fusidicates; fatty acid derivatives of amino acids, oligopeptides, and polypeptides; glyceride derivatives of amino acids, oligopeptides, and polypeptides; lecithin and hydrogenated lecithin; lysolecithin and hydrogenated lysolecithin; phospholipids and their derivatives; lysophospholipids and their derivatives; carnitine fatty acid ester salts; alkyl sulfates; fatty acid salts; sodium doxate; acyl lactylates; mono- and diacetylated tartaric acid esters of mono- and di-glycerides; succinylated mono- and di-glycerides; citrate esters of mono- and di-glycerides; and mixtures thereof.

[0308] Examples of ionic surfactants among the aforementioned group include lecithin, lysolecithin, phospholipids, lysophospholipids and their derivatives; carnitine fatty acid ester salts; alkyl sulfates; fatty acid salts; sodium doxate; acyl lactylates; mono- and diacetylated tartaric acid esters of mono- and di-glycerides; succinylated mono- and di-glycerides; citrate esters of mono- and di-glycerides; and mixtures thereof.

[0309] Ionic surfactants include lecithin, lysolecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylserine, lysophosphatidylcholine, lysophosphatidiethanolamine, lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylserine, PEG-phosphatidiethanolamine, PVP-phosphatidiethanolamine, fatty acid lactyl esters, stearoyl-2-lactylate, stearoyl lactate, and sucrose. Synylated monoglycerides, mono / diacetylated tartrate esters of mono / diglycerides, citrate esters of mono / diglycerides, cholyl sarcosine, caproate, caprylate, caprate, laurate, myristate, palmitate, oleate, ricinoleate, linoleate, linolenate, stearate, lauryl sulfate, teracecil sulfate, doxate, lauroyl carnitine, palmitoyl carnitine, myristoyl carnitine, and ionized forms of their salts and mixtures.

[0310] Examples of hydrophilic nonionic surfactants, though not limited to them, include: alkyl glucosides; alkyl maltosides; alkyl thioglucosides; lauryl macrogol glycerides; polyoxyalkylene alkyl ethers such as polyethylene glycol alkyl ethers; polyoxyalkylene alkylphenols such as polyethylene glycol alkylphenols; polyoxyalkylene alkylphenol fatty acid esters such as polyethylene glycol fatty acid monoesters and polyethylene glycol fatty acid diesters; polyethylene glycol glycerol fatty acid esters; polyglycerol fatty acid esters; polyoxyalkylene sorbitan fatty acid esters such as polyethylene glycol sorbitan fatty acid esters; hydrophilic transesterifications of polyols with at least one component from the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids, and sterols; polyoxyethylene sterols, their derivatives and analogs; polyoxyethylated vitamins and their derivatives; polyoxyethylene-polyoxypropylene block copolymers; and mixtures thereof; polyethylene glycol sorbitan fatty acid esters, and hydrophilic transesterifications of polyols with at least one component from the group consisting of triglycerides, vegetable oils, and hydrogenated vegetable oils. The polyol may be glycerol, ethylene glycol, polyethylene glycol, sorbitol, propylene glycol, pentaerythritol, or sugars.

[0311] Other hydrophilic nonionic surfactants include, but are not limited to, PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-32 dilaurate, PEG-12 oleate, PEG-15 oleate, PEG-20 oleate, PEG-20 dioleate, PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32 distearate, PEG-40 stearate, PEG-100 stearate, PEG-20 dilaurate, PEG-25 glyceryl trioleate, PEG-32 dioleate, PEG-20 glyceryl laurate, PEG-30 glyceryl laurate, PEG-20 glyceryl stearate, PEG-20 glyceryl oleate, PEG-30 glyceryl oleate, PEG-30 glyceryl laurate, and PEG-40 glyceryl laurate. PEG-40 Palm Kernel Oil, PEG-50 Hydrogenated Castor Oil, PEG-40 Castor Oil, PEG-35 Castor Oil, PEG-60 Castor Oil, PEG-40 Hydrogenated Castor Oil, PEG-60 Hydrogenated Castor Oil, PEG-60 Corn Oil, PEG-6 (Caprylic / Capric Acid) Glycerides, PEG-8 (Caprylic / Capric Acid) Glycerides, Polyglyceryl-10 Laurate, PEG-30 Cholesterol, PEG-25 Phytosterol, PEG-30 Soybean Steroid Examples include oleate, PEG-20 trioleate, PEG-40 sorbitan oleate, PEG-80 sorbitan laurate, polysorbate 20, polysorbate 80, POE-9 lauryl ether, POE-23 lauryl ether, POE-10 oleyl ether, POE-20 oleyl ether, POE-20 stearyl ether, tocopheryl succinate PEG-100, PEG-24 cholesterol, polyglyceryl-10 oleate, Tween 40, Tween 60, sucrose monostearate, sucrose monolaurate, sucrose monopalmitate, PEG-10-100 nonylphenol series, PEG-15-100 octylphenol series, and poloxamer.

[0312] Suitable lipophilic surfactants include, but are not limited to, aliphatic alcohols; glycerol fatty acid esters; acetylated glycerol fatty acid esters; lower alcohol fatty acid esters; propylene glycol fatty acid esters; sorbitan fatty acid esters; polyethylene glycol sorbitan fatty acid esters; sterols and sterol derivatives; polyoxyethylated sterols and sterol derivatives; polyethylene glycol alkyl ethers; sugar esters; sugar ethers; lactic acid derivatives of mono- and di-glycerides; hydrophobic transesterifications of polyols with at least one component from the group consisting of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids, and sterols; oil-soluble vitamins / vitamin derivatives; and mixtures thereof. Of this group, preferred lipophilic surfactants include glycerol fatty acid esters, propylene glycol fatty acid esters, and mixtures thereof, or hydrophilic transesterifications of polyols with at least one component from the group consisting of vegetable oils, hydrogenated vegetable oils, and triglycerides.

[0313] In one embodiment, the composition may contain a solubilizer to ensure good solubilization and / or dissolution of the compounds of the Disclosure and to minimize precipitation of the compounds of the Disclosure. This may be particularly important for compositions for parenteral use, such as compositions for injection. Solubilizers may be added to increase the solubility of other components, such as hydrophilic drugs and / or surfactants, or to maintain the composition as a stable or homogeneous solution or dispersion.

[0314] Examples of suitable solubilizers include, but are not limited to, alcohols and polyols such as ethanol, isopropanol, butanol, benzyl alcohol, ethylene glycol, propylene glycol, butanediol and their isomers, glycerol, pentaerythritol, sorbitol, mannitol, transktol, dimethyl isosorbide, polyethylene glycol, polypropylene glycol, polyvinyl alcohol, hydroxypropyl methylcellulose and other cellulose derivatives, cyclodextrin and cyclodextrin derivatives; polyethylene glycol ethers having an average molecular weight of about 200 to about 6000, such as tetrahydrofurfuryl alcohol PEG ether (glycoflor) or methoxyPEG; 2-pyrrolidone, 2-piperidone, ε-caprolactam, N-al Amides and other nitrogen-containing compounds such as chilpyrrolidone, N-hydroxyalkylpyrrolidone, N-alkylpiperidone, N-alkylcaprolactam, dimethylacetamide, and polyvinylpyrrolidone; esters such as ethyl propionate, tributyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, triethyl citrate, ethyl oleate, ethyl caprylate, ethyl butyrate, triacetin, propylene glycol monoacetate, propylene glycol diacetate, ε-caprolactone and its isomers, δ-valerolactone and its isomers, β-butyllactone and its isomers; and other solubilizers known in the art such as dimethylacetamide, dimethyl isosorbide, N-methylpyrrolidone, monooctanoin, diethylene glycol monoethyl ether, and water.

[0315] A mixture of solubilizers may be used. Examples, but not limited to, include triacetin, triethyl citrate, ethyl oleate, ethyl caprylate, dimethylacetamide, N-methylpyrrolidone, N-hydroxyethylpyrrolidone, polyvinylpyrrolidone, hydroxypropyl methylcellulose, hydroxypropyl cyclodextrin, ethanol, polyethylene glycol 200-100, glycoflore, transktol, propylene glycol, and dimethyl isosorbide. Particularly preferred solubilizers include sorbitol, glycerol, triacetin, ethyl alcohol, PEG-400, glycoflore, and propylene glycol.

[0316] The amount of solubilizer that may be included is not particularly limited. A given amount of solubilizer may be limited to a bioacceptable amount, which can be easily determined by those skilled in the art. In some situations, for example, to maximize the concentration of the drug, it may be advantageous to include an amount of solubilizer far exceeding the bioacceptable amount, and the excess solubilizer is removed by conventional techniques such as distillation or evaporation before the composition is provided to the patient. If present, the solubilizer may be present in a weight ratio of 10% by weight, 25% by weight, 50% by weight, 100% by weight, or up to about 200% by weight, based on the combined weight of the drug and other excipients. If desired, very small amounts of solubilizer, such as 5%, 2%, 1%, or less, may be used. Typically, the solubilizer may be present in amounts of about 1% by weight to about 100% by weight, more typically about 5% by weight to about 25% by weight.

[0317] The composition may further contain one or more pharmaceutically acceptable additives and excipients. Such additives and excipients include, but are not limited to, antifogging agents, defoaming agents, buffering agents, polymers, antioxidants, preservatives, chelating agents, viscosity modifiers, isotonic agents (tonicizers), flavoring agents, colorants, odorants, opacifiers, suspending agents, binders, fillers, plasticizers, lubricants, and mixtures thereof.

[0318] In addition, acids or bases may be incorporated into the composition to facilitate processing, improve stability, or for other reasons. Examples of pharmaceutically acceptable bases include amino acids, amino acid esters, ammonium hydroxide, potassium hydroxide, sodium hydroxide, sodium bicarbonate, aluminum hydroxide, calcium carbonate, magnesium hydroxide, magnesium aluminum silicate, synthetic aluminum silicate, synthetic hydrocalcite, magnesium aluminum hydroxide, diisopropylethylamine, ethanolamine, ethylenediamine, triethanolamine, triethylamine, triisopropanolamine, trimethylamine, tris(hydroxymethyl)aminomethane (TRIS), and similar. Furthermore, bases that are salts of pharmaceutically acceptable acids such as acetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonic acid, amino acids, ascorbic acid, benzoates, boric acid, butyric acid, carbonic acid, citric acid, fatty acids, formic acid, fumaric acid, gluconic acid, hydroquinonesulfonic acid, isoascorbic acid, lactic acid, maleic acid, oxalic acid, para-bromophenylsulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, uric acid, and the like are also suitable. Salts of polybasic acids such as sodium phosphate, disodium hydrogen phosphate, and sodium dihydrogen phosphate can also be used. When the base is a salt, the cation can be any convenient and pharmaceutically acceptable cation such as ammonium, alkali metals, alkaline earth metals, and the like. Examples, but not limited to, include sodium, potassium, lithium, magnesium, calcium, and ammonium.

[0319] Suitable acids are pharmaceutically acceptable organic or inorganic acids. Examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, boric acid, phosphoric acid, and similar substances. Examples of suitable organic acids include acetic acid, acrylic acid, adipic acid, alginic acid, alkanesulfonic acid, amino acids, ascorbic acid, benzoates, boric acid, butyric acid, carbonic acid, citric acid, fatty acids, formic acid, fumaric acid, gluconic acid, hydroquinonesulfonic acid, isoascorbic acid, lactic acid, maleic acid, methanesulfonic acid, oxalic acid, para-bromophenylsulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, uric acid, and similar substances. [Examples]

[0320] In the examples, the following abbreviations are used: ATP = adenosine triphosphate, DBU = 1,8-diazabicyclo[5.4.0]undeca-7-ene, DCE = 1,2-dichloroethane, DCM = dichloromethane, DEA = diethylamine, DHP = dihydropyran, DIPEA = N,N-diisopropylethylamine, DME = dimethoxyethane, DMF = dimethylformamide, DMSO = dimethyl sulfoxide, EtOH = ethanol, DTT = dithiothreitol, HPLC = high-performance liquid chromatography, PMB = para-methoxybenzyl, PPTS = pyridinium p-toluenesulfonate, SFC = supercritical fluid chromatography, TBME = tert-butylmethyl ether, TEA = triethylamine, TFA = trifluoroacetic acid, THF = tetrahydrofuran, THP = tetrahydropyran.

[0321] All chemicals, reagents, and solvents were purchased from commercial sources where available and used without further purification.

[0322] Example 1 Synthesis of 4-(((2'-(((1R,4R)-4-aminocyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (intermediate 1)

[0323] [ka]

[0324] Step 1: Preparation of tetrahydro-4H-pyran-4,4-dicarbonitride (INT-2)

[0325] [ka]

[0326] To a solution of 1-bromo-2-(2-bromoethoxy)ethane (INT-1, 20 g, 86.24 mmol) and propanedinitrile (6.27 g, 94.86 mmol) in DMF (30 mL), DBU (26.26 g, 172.48 mmol) was added. The reaction mixture was stirred at 85 °C for 3 hours, cooled to ambient temperature, diluted with water (100 mL), and extracted with ethyl acetate (100 mL x 3). The combined organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain tetrahydro-4H-pyran-4,4-dicarbonitride (INT-2, 12.61 g, crude) as a brown solid. The crude product was used directly in the next step without further purification. 1 ¹H NMR (400 MHz, methanol-d4): δ = 3.86–3.77 (m, 4H), 2.32–2.21 (m, 4H).

[0327] Step 2: Preparation of 4-(aminomethyl)tetrahydro-2H-pyran-4-carbonitrile (INT-3)

[0328] [ka]

[0329] To a solution of tetrahydro-4H-pyran-4,4-dicarbonitride (INT-2, 9.0 g, 66.10 mmol) in EtOH (270 mL), NaBH4 (7.50 g, 198.31 mmol) was added in small increments. The reaction mixture was stirred at 20°C for 4 hours, quenched with water (200 mL), and extracted with ethyl acetate (200 mL x 3). The combined organic phase was washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain 4-(aminomethyl)tetrahydro-2H-pyran-4-carbonitride (INT-3, 7.23 g, yield 78%) as a brown oil. The crude product was used directly in the next step without further purification.

[0330] Step 3: Preparation of 4-(((6-bromopyridine-2-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (INT-4)

[0331] [ka]

[0332] TEA (13.05 g, 128.94 mmol) was added to a solution of 4-(aminomethyl)tetrahydro-2H-pyran-4-carbonitrile (INT-3, 7.23 g, 51.58 mmol) and 2-bromo-6-fluoropyridine (7.72 g, 43.84 mmol) in DMSO (80 mL). The reaction mixture was stirred at 130 °C for 18 hours, cooled to ambient temperature, diluted with ethyl acetate (100 mL), washed with saturated NaHCO3 solution and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The resulting residue was purified by silica gel chromatography (Biotage 20g Silica Flash Column; 0-25% petroleum ether in ethyl acetate @ 40 mL / min) to obtain 4-(((6-bromopyridine-2-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (INT-4, 3.8 g, yield 18.35%) as a pale green solid. MS (ESI) m / z = 296.1 [M+H] + .

[0333] Step 4: Preparation of 4-(((5'-chloro-2'-fluoro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (INT-5)

[0334] [ka]

[0335] 4-(((6-bromopyridine-2-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (INT-4, 3.8 g, 12.83 mmol), (5-chloro-2-fluoro-4-pyridyl)boronic acid (3.37 g, 19.25 mmol), and Pd(dppf)Cl2 (938.84 mg, 1.28 mmol) were dissolved in DME (40 mL), to which Na2CO3 (2 M, 16.04 mL) was added. The reaction mixture was sealed, stirred at 110°C under N2 for 4 hours, cooled to ambient temperature, diluted with water (40 mL), and extracted with ethyl acetate (70 mL x 3). The combined organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The resulting residue was purified by silica gel chromatography (Biotage 40g Silica Flash Column; 20-26% petroleum ether in ethyl acetate @ 80 mL / min) to obtain 4-(((5'-chloro-2'-fluoro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (INT-5, 3.6 g, yield 73%) as a yellow oil. MS (ESI) m / z = 347.1 [M+H] + .

[0336] Step 5: Preparation of 4-(((2'-(((1R,4R)-4-aminocyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (intermediate 1)

[0337] [ka]

[0338] TEA (2.53 g, 24.97 mmol) was added to a solution of 4-(((5'-chloro-2'-fluoro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (INT-5, 4.33 g, 12.49 mmol) and cyclohexane-1,4-diamine (2.14 g, 18.73 mmol) in DMSO (50 mL). The reaction mixture was stirred at 110 °C for 16 hours, diluted with water (40 mL), and extracted with ethyl acetate (100 mL x 3). The combined organic phase was washed with brine (60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The resulting residue was dissolved in ethyl acetate (100 mL), added dropwise to hydrochloric acid / dioxane (50 mL), filtered, and washed with ethyl acetate. The resulting solid was dissolved in water (150 mL), basicized to pH 9 with NaHCO3, and extracted with ethyl acetate (100 mL x 3). The combined organic phase was washed with brine (60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain 4-(((2'-(((1R,4R)-4-aminocyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (intermediate 1, 4.1 g, yield 61.11%) as a brown solid. MS (ESI) m / z = 441.3 [M+H] + .

[0339] Example 2 Synthesis of 1-((2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazole-5-yl)methoxy)propan-2-one (intermediate 2)

[0340] [ka]

[0341] Step 1: Preparation of 5-(chloromethyl)-2H-tetrazole (INT-7)

[0342] [ka]

[0343] NaN3 (4.87 g, 74.97 mmol) was added to a solution of AlCl3 (3.67 g, 27.55 mmol) in THF (50 mL). The reaction mixture was stirred at 60 °C for 2 hours and then cooled to 20 °C. 2-chloroacetonitrile (INT-6, 2.0 g, 26.49 mmol) was added, and the reaction mixture was heated to 70 °C and stirred for 24 hours. The reaction mixture was concentrated, and the resulting residue was acidified to pH 2 with 37% hydrochloric acid solution and extracted with siRNA (100 mL x 3). The combined organic layers were dried over Na2SO4, filtered, and concentrated to obtain 5-(chloromethyl)-2H-tetrazole (INT-7, 3.0 g, crude) as a white solid. The crude product was used directly in the next step without further purification. 1 H NMR (400 MHz, DMSO-d6): δ = 5.08 (s, 2H).

[0344] Step 2: Preparation of 5-(chloromethyl)-2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazole (INT-8)

[0345] [ka]

[0346] PPTS (203.54 mg, 0.809 mmol) was added to a solution of 5-(chloromethyl)-2H-tetrazole (INT-7, 3.0 g, 25.31 mmol) and DHP (4.26 g, 50.62 mmol) in acetone (100 mL). The reaction mixture was stirred at 45 °C for 3 hours and then concentrated. The residue was purified by silica gel chromatography (Biotage 20 g Silica Flash Column; 0-10% petroleum ether in ethyl acetate @ 40 mL / min) to obtain 5-(chloromethyl)-2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazole (INT-8, 4.6 g, yield 89%) as a colorless oil. 1H NMR (400 MHz, DMSO-d6): δ = 6.18 (dd, J = 8.0, 2.8 Hz, 1H), 5.06 (s, 2H), 3.81-3.76 (m, 2H), 2.51-2.26 (m, 1H), 2.25-2.13 (m, 1H), 2.12-1.98 (m, 1H), 1.64-1.62 (m, 1H), 1.61-1.601 (m, 2H).

[0347] Step 3: Preparation of 5-(((2-methylallyl)oxy)methyl)-2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazole (INT-9)

[0348] [ka]

[0349] To a solution of 2-methylpropa-2-en-1-ol (1.07 g, 14.80 mmol, 1.25 mL) in DMF (2 mL), NaH (789 mg, 19.74 mmol, 60% purity) was added, and the reaction mixture was stirred at 0°C for 0.5 hours. 5-(chloromethyl)-2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazole (INT-8, 2.0 g, 9.87 mmol) was added, and the reaction mixture was stirred at 25°C for 2 hours. The mixture was then quenched with brine (50 mL) and extracted with siRNA (50 mL x 3). The combined organic layers were washed with brine (50 mL x 7), dried over Na2SO4, filtered, and concentrated to obtain 5-(((2-methylallyl)oxy)methyl)-2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazole (INT-9, 2.3 g, 97% yield) as a white solid. The crude product was used directly in the next step without further purification. 1H NMR (400 MHz, DMSO-d6): δ = 6.15 (dd, J = 8.0, 3.2 Hz, 1H), 4.95 (s, 1H), 4.89 (s, 1H), 4.71 (s, 2H), 3.95 (s, 2H), 3.82-3.75 (m, 2H), 2.26-2.25 (m, 1H), 2.25-2.24 (m, 1H), 2.16-2.10 (m, 1H), 1.67-1.66 (m, 1H), 1.64 (s, 3H), 1.62-1.58 (m, 2H).

[0350] Step 4: Preparation of 1-((2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazole-5-yl)methoxy)propan-2-one (intermediate 2)

[0351] [ka]

[0352] 5-(((2-methylallyl)oxy)methyl)-2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazole (INT-9, 1.0 g, 4.20 mmol) and K2OsO4·2H2O (15.46 mg, 0.042 mmol) were added dropwise to a solution of NaIO4 (2.06 g, 9.65 mmol) in H2O (8 mL) in a solution of K2OsO4·2H2O (15.46 mg, 0.042 mmol) in H2O (3.6 mL) and THF (4 mL). The reaction mixture was stirred at 25°C for 12 hours, filtered, and extracted with siRNA (30 mL × 5). The combined organic layer was dried over Na2SO4, filtered, and concentrated to obtain 1-((2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazole-5-yl)methoxy)propan-2-one (intermediate 2, 800 mg, yield 79%) as a yellow oil. 1H NMR (400 MHz, DMSO-d6): δ = 6.17 (dd, J = 8.0, 3.6 Hz, 1H), 5.06 (s, 2H), 3.84-3.73 (m, 3H), 2.24-2.15 (m, 1H), 2.15-2.13 (m, 1H), 1.99-1.75 (m, 1H), 1.74-1.72 (m, 4H), 1.64-1.59 (m, 3H). MS (ESI) m / z = 263.3 [M+Na] + .

[0353] Example 3 Synthesis of ethyl 2-(2-oxopropoxy)acetate (intermediate 3)

[0354] [ka]

[0355] Step 1: Preparation of ethyl 2-((2-methylallyl)oxy)acetate (INT-11)

[0356] [ka]

[0357] To a solution of 2-methylpropa-2-en-1-ol (1.0 g, 13.87 mmol) in DMF (12 mL), NaH (610 mg, 15.26 mmol) was added. The reaction mixture was stirred at 0°C for 10 minutes, and ethyl 2-bromoacetate (INT-10, 2.32 g, 13.87 mmol) was added. The reaction mixture was stirred at 0°C for 1 hour, quenched with water (5 mL) at 0°C, and extracted with ethyl acetate (20 mL x 3). The combined organic phase was washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The resulting residue was purified by silica gel flash column chromatography (0-20% ethyl acetate in petroleum ether) to obtain ethyl 2-((2-methylallyl)oxy)acetate (INT-11, 780 mg, yield 36%) as a colorless liquid. 1H NMR (400 MHz, DMSO-d6) δ = 4.89-4.86 (m, 1H), 4.57-4.45 (m, 1H), 4.11 (s, 2H), 4.09-4.06 (m, 2H), 3.92-3.90 (m, 2H), 1.68-1.67 (m, 3H), 1.23-1.16 (m, 3H). MS (ESI) m / z = 159.3 [M+H] + .

[0358] Step 2: Preparation of ethyl 2-(2-oxopropoxy)acetate (intermediate 3)

[0359] [ka]

[0360] To a solution of ethyl 2-((2-methylallyl)oxy)acetate (INT-11, 1.5 g, 9.48 mmol) in THF (8 mL), K2OsO4·2H2O (34.94 mg, 0.1 mmol) in a mixture of H2O (4 mL) and THF (9 mL), and NaIO4 (4.66 g, 21.81 mmol) in H2O (12 mL) were added. The reaction mixture was stirred at 20°C for 11 hours and concentrated under vacuum. The resulting residue was purified by silica gel chromatography (Biotage 20 g Silica Flash Column; 0-10% petroleum ether in ethyl acetate @ 40 mL / min) to obtain ethyl 2-(2-oxopropoxy)acetate (intermediate 3, 850 mg, yield 56%) as a colorless liquid. 1 H NMR (400 MHz, CDCl3) δ = 4.24-4.14 (m, 6H), 2.17 (s, 3H), 1.27 (t, J = 7.2 Hz, 3H).

[0361] Example 4 Synthesis of 1-(1-(4-methoxybenzyl)-1H-tetrazole-5-yl)propan-2-one (intermediate 4)

[0362] [ka]

[0363] Step 1: Preparation of 4-hydrazine-6-methylpyrimidine (INT-13)

[0364] [ka]

[0365] To a solution of 4-chloro-6-methylpyrimidine (INT-12, 2.5 g, 19.45 mmol) in dioxane (30 mL), NH2NH2·H2O (1.67 g, 33.45 mmol) and K2CO3 (2.74 g, 19.84 mmol) were added. The reaction mixture was stirred at 100 °C for 6 hours, diluted with H2O (20 mL), and extracted with siRNA (50 mL x 3). The combined organic layer was washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain 4-hydrazine-6-methylpyrimidine (INT-13, 2 g, 16.11 mmol, yield 82.85%) as a yellow solid. The crude product was used directly in the next step without further purification.

[0366] Step 2: Preparation of (Z)-N-(1-(1H-tetrazole-5-yl)prop-1-en-2-yl)formamide (INT-14)

[0367] [ka]

[0368] To a solution of 4-hydrazine-6-methylpyrimidine (INT-13, 0.9 g, 7.25 mmol) in H2O (2 mL), HCl (2 M, 3.62 mL) and NaNO2 (500.19 mg, 7.25 mmol) were added. The reaction mixture was stirred at 0°C for 8 hours, filtered, and the filtered cake was concentrated under reduced pressure to obtain (Z)-N-(1-(1H-tetrazole-5-yl)propa-1-en-2-yl)formamide (INT-14, 0.6 g, 3.92 mmol, yield 54.04%) as a brown solid. The crude product was used directly in the next step without further purification. MS (ESI) m / z = 296.1 [M+H] + .

[0369] Step 3: Preparation of 5-(2-oxopropyl)-1H-tetrazole-1-ium chloride (INT-15)

[0370] [ka]

[0371] (Z)-N-(1-(1H-tetrazole-5-yl)propa-1-en-2-yl)formamide (INT-14, 0.8 g, 5.22 mmol) was dissolved in H2O (4 mL), to which HCl (1 mL) was added. The reaction mixture was stirred at 80 °C for 12 hours and concentrated under reduced pressure to obtain 5-(2-oxopropyl)-1H-tetrazole-1-ium chloride (INT-15, 0.7 g, 4.31 mmol, yield 82.42%) as a yellow solid. The crude product was used directly in the next step without further purification.

[0372] Step 5: Preparation of 1-(1-(4-methoxybenzyl)-1H-tetrazole-5-yl)propan-2-one (intermediate 4)

[0373] [ka]

[0374] 5-(2-oxopropyl)-1H-tetrazole-1-ium chloride (INT-15, 600 mg, 4.76 mmol, 1 equivalent) and 1-(chloromethyl)-4-methoxybenzene (1.12 g, 7.14 mmol, 971.83 uL, 1.5 equivalents) were dissolved in THF (15 mL) and DIPEA (3.07 g, 23.79 mmol, 4.14 mL, 5 equivalents) was added. The reaction mixture was stirred at 45 °C for 16 hours, diluted with water (20 mL), and extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (40 mL), dried over Na₂SO₄, filtered, and concentrated. The resulting residue was purified by flash silica gel chromatography (ISCO®; 20g SepaFlash® Silica Flash Column, 0-6% methanol / dichloromethane @ 35 mL / min) to obtain 1-(1-(4-methoxybenzyl)-1H-tetrazole-5-yl)propan-2-one (intermediate 4, 440 mg, 1.14 mmol, yield 24.04%, purity 64%) as a pale yellow oil. MS (ESI) m / z = 247.2 [M+H] + .

[0375] Example 5 Synthesis of 4-(((2'-(((1R,4R)-4-(((R)-1-((2H-tetrazole-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 2) and 4-(((2'-(((1S,4R)-4-(((S)-1-((2H-tetrazole-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 3)

[0376] [ka]

[0377] 4-(((2'-(((1R,4R)-4-aminocyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (intermediate 1, 500 mg, 1.13 mmol) and 1-((2-(tetrahydro-2H-pyran-2-yl)-2H-tetrazole-5-yl)methoxy)propan-2-one (intermediate 2, 354 mg, 1.47 mmol) were dissolved in AcOH (204 mg, 3.40 mmol) and DCE (15 mL), to which NaBH(OAc)3 (336 mg, 1.59 mmol) was added at 0°C. The mixture was stirred at 25°C for 12 hours, quenched with MeOH (5 mL), and concentrated. The resulting residue was purified by preparative HPLC (Phenomenex Luna C18 75×30mm (particle size 3μm); 0-35% acetonitrile / water (0.225% FA); 35 min; 25 mL / min) to obtain 4-(((2'-((4-((1-((2H-tetrazole-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 1, 140 mg, yield 21%) as a white solid. MS (ESI) m / z = 581.5 [M+H] + .

[0378] 4-(((2'-((4-((1-((2H-tetrazole-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 1) is processed into chiral SFC (Chiralpak OD-3 100×4.6mm (particle size 3μm); 5~40% EtOH (0.05%) Separation by DEA / CO2; 8 min; 2.8 mL / min) to 4-(((2'-(((1R,4R)-4-(((R)-1-((2H-tetrazole-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 2, 35 mg, yield 35%, pure We obtained compound 3, 35 mg, yield 35%, purity 99%, ee 97%, and compound 4-(((2'-(((1S,4R)-4-(((S)-1-((2H-tetrazole-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 3, 35 mg, yield 35%, purity 99%, ee 97%).

[0379] Compound 2: 11H NMR (400 MHz, CDCl3): δ = 8.04 (s, 1H), 7.48 (t, J = 7.6 Hz, 1H), 6.91 (d, J = 7.6 Hz, 1H), 6.54 (s, 1H), 6.49 (d, J = 8.0 Hz, 2H), 5.09 - 5.05 (m, 1H), 4.87 - 4.83 (m, 2H), 4.45 (d, J = 8.0 Hz, 1H), 3.97 - 3.95 (m, 2H), 3.87 - 3.85 (m, 1H), 3.74 - 3.63 (m, 6H), 3.57 - 3.55 (m, 2H), 3.22 - 3.21 (m, 1H), 2.41 - 2.40 (m, 2H), 2.30 - 2.10 (m, 2H), 1.91 - 1.77 (m, 4H), 1.76 - 1.70 (m, 2H), 1.50 - 1.48 (m, 3H), 1.29 - 1.26 (m, 2H). MS (ESI) m / z = 581.5 [M+H] + .

[0380] Compound 3: 1 1H NMR (400 MHz, CDCl3): δ = 8.05 (s, 1H), 7.48 (t, J = 7.6 Hz, 1H), 6.91 (d, J = 7.6 Hz, 1H), 6.54 (s, 1H), 6.49 (d, J = 8.0 Hz, 2H), 5.08 - 5.05 (m, 1H), 4.89 - 4.83 (m, 2H), 4.45 (d, J = 8.0 Hz, 1H), 3.97 - 3.95 (m, 2H), 3.87 - 3.85 (m, 1H), 3.74 - 3.63 (m, 6H), 3.57 - 3.55 (m, 2H), 3.22 - 3.21 (m, 1H), 2.41 - 2.40 (m, 2H), 2.30 - 2.10 (m, 2H), 1.91 - 1.77 (m, 4H), 1.76 - 1.70 (m, 2H), 1.50 - 1.48 (m, 3H), 1.29 - 1.26 (m, 2H). MS (ESI) m / z = 581.5 [M+H] + .

[0381] Example 6 Synthesis of ethyl 2-((R)-2-(((1R,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4]-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetate (compound 5) and ethyl 2-((S)-2-(((1R,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4]-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetate (compound 6)

[0382] [ka]

[0383] 4-(((2'-(((1R,4R)-4-aminocyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (intermediate 1, 200 mg, 0.45 mmol), ethyl 2-(2-oxopropoxy)acetate (intermediate 3, 80 mg, 0.5 mmol), and AcOH (82 mg, 1.36 mmol) were dissolved in DCE (2 mL), to which NaBH(OAc)3 (134 mg, 0.63 mmol) was added at 0°C. The reaction mixture was stirred at 20°C for 20 hours, quenched with water (10 mL) at 0°C, and extracted with ethyl acetate (30 mL x 3). The combined organic phase was washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The resulting residue was purified by preparative HPLC (Phenomenex Luna C18 100×30mm (particle size 5μm); 10-40% acetonitrile / water (0.225% FA); 10 min; 25 mL / min) to obtain ethyl 2-(2-((4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetate (compound 4) as a white solid.

[0384] Ethyl 2-(2-((4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetate (compound 4) is processed into chiral SFC (Chiralpak AD-3 100×4.6mm (particle size 3μm); 5~40% EtOH (0.05%) Further separation by DEA / CO2 (8 min; 2.8 mL / min) yielded ethyl 2-((R)-2-(((1R,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4]-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetate (compound 5, 18.8 mg, yield 7%, purity >9 9%, ee>99% and ethyl 2-((S)-2-(((1R,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4]-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetate (compound 6, 21.8 mg, yield 8%, purity 99%, ee 90%) were obtained as white solids.

[0385] Compound 5: 11H NMR (400 MHz, CDCl3) δ = 8.11 (s, 1H), 7.53 (t, J = 8.0 Hz, 1H), 6.97 (d, J = 8.0 Hz, 1H), 6.55 (s, 1H), 6.51 (d, J = 8.0 Hz, 1H), 4.78 (t, J = 4.0 Hz, 1H), 4.43 (d, J = 8.0 Hz, 1H), 4.30 - 4.17 (m, 2H), 4.11(s, 2H), 4.05 - 3.97 (m, 2H), 3.81 - 3.76 (m, 2H), 3.74 - 3.66 (m, 2H), 3.61 - 3.49 (m, 2H), 3.47 - 3.34 (m, 1H), 3.19 - 3.01 (m, 1H), 2.82 - 2.47 (m, 1H), 2.22 - 2.13 (m, 2H), 2.11 - 2.00 (m, 2H), 1.97 - 1.90 (m, 2H), 1.80 - 1.72 (m, 2H), 1.32 - 1.20 (m, 7H), 1.15 - 1.06 (m, 3H). MS (ESI) m / z = 585.4 [M+H] + .

[0386] Compound 6: 1 1H NMR (400 MHz, CDCl3) δ = 8.44 (s, 1H), 8.10 (s, 1H), 7.52 (t, J = 8.0 Hz, 1H), 6.97 (d, J = 4.0 Hz, 1H), 6.57 (s, 1H), 6.52 (d, J = 8.0 Hz, 1H), 4.82 (t, J = 8.0 Hz, 1H), 4.28 - 4.20 (m, 2H), 4.17 - 4.11 (m, 2H), 4.03 - 3.97 (m, 2H), 3.81 - 3.64 (m, 7H), 3.31 - 3.27 (m, 1H), 3.02 - 2.92 (m, 1H), 2.28 - 2.11 (m, 4H), 1.97 - 1.89 (m, 2H), 1.81 - 1.72 (m, 2H), 1.67 - 1.46 (m, 2H), 1.33 - 1.25 (m, 8H). MS (ESI) m / z = 585.4 [M+H] + .

[0387] Example 7 Synthesis of 2-((R)-2-(((1R,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetic acid (compound 8) and 2-((S)-2-(((1R,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetic acid (compound 9)

[0388] [ka]

[0389] NaOH (143 mg, 3.59 mmol) was added to a solution of ethyl 2-(2-((4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetate (compound 4, 210 mg, 0.35 mmol) in THF (3 mL) and H2O (1 mL). The reaction mixture was stirred at 25°C for 2 hours. The aqueous phase was acidified to pH 4 with aqueous HCl (1N) and concentrated under vacuum. The resulting residue was dissolved in DMF, filtered, concentrated under vacuum, and purified by preparative HPLC (Phenomenex Luna C18 100×40mm (particle size 3μm); 15~45% acetonitrile / water (0.225% FA); 10 min; 25 mL / min) to obtain 2-(2-((4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetic acid (compound 7).

[0390] 2-(2-((4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetic acid (compound 7) is processed into chiral SFC (Chiralpak AD-3 100×4.6mm (particle size 3μm); 5~40% EtOH (0.05%) Further separation by DEA / CO2 (8 min; 2.8 mL / min) yielded 2-((R)-2-(((1R,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetic acid (compound 8, 32.5 mg, yield 16%, purity 98%). %, ee>99%) and 2-((S)-2-(((1R,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetic acid (compound 9, 34.8 mg, yield 17%, purity 99%, ee>99%) were obtained as a yellow solid.

[0391] Compound 8: 1 H NMR (400 MHz, DMSO-d6) δ = 8.16 (s, 1H), 8.03 (s, 1H), 7.54-7.45 (m, 1H), 7.12-7.05 (m, 1H), 6.79-6.70 (m, 2H), 6.68-6.60 (m, 2H), 3.92-3.86 (m, 2H), 3.71-3.62 (m, 7H), 3.49-3.46 (m, 2H), 3.04-2.92 (m, 2H), 2.13-2.00 (m, 4H), 1.87-1.80 (m, 2H), 1.71-1.62 (m, 2H), 1.54-1.44 (m, 2H), 1.30-1.21 (m, 2H), 1.17-1.11 (m, 3H). MS (ESI) m / z = 557.3 [M+H] + .

[0392] Compound 9: 1H NMR (400 MHz, DMSO-d6) δ = 8.17 (s, 1H), 8.03 (s, 1H), 7.54-7.45 (m, 1H), 7.13-7.04 (m, 1H), 6.78-6.71 (m, 2H), 6.69-6.64 (m, 1H), 6.63-6.60 (m, 1H), 3.92-3.86 (m, 2H), 3.70-3.61 (m, 7H), 3.50-3.46 (m, 2H), 3.04-2.90 (m, 2H), 2.14-2.02 (m, 4H), 1.88-1.80 (m, 2H), 1.73-1.62 (m, MS (ESI) m / z = 557.3 [M+H] + .

[0393] Example 8 Synthesis of 4-(((2'-(((1R,4R)-4-(((R)-1-(1H-tetrazole-5-yl)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 12) and 4-(((2'-(((1S,4R)-4-(((S)-1-(1H-tetrazole-5-yl)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 13)

[0394] [ka]

[0395] Step 1: Preparation of 4-(((5'-chloro-2'-(((1R,4R)-4-((1-(1-(4-methoxybenzyl)-1H-tetrazole-5-yl)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (Compound 10)

[0396] [ka]

[0397] AcOH (137.34 mg, 2.29 mmol, 130.80 uL, 2 equivalents) and NaBH(OAc)3 (727.05 mg, 3.43 mmol, 3 equivalents) were added to a solution of 4-(((2'-(((1R,4R)-4-aminocyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (intermediate 1, 504.24 mg, 1.14 mmol, 1 equivalent) and 1-(1-(4-methoxybenzyl)-1H-tetrazole-5-yl)propan-2-one (intermediate 4, 440 mg, 1.14 mmol, purity 64%, 1 equivalent) in DCE (20 mL). The reaction mixture was stirred at 20°C for 16 hours, diluted with water (40 mL), basicized to pH 7-8 with saturated NaHCO3, and extracted with DCM (60 mL x 3). The combined organic layers were washed with brine (80 mL), dried over Na2SO4, filtered, and concentrated. The resulting residue was purified by flash silica gel chromatography (20g SepaFlash® Silica Flash Column, 0-8% methanol / dichloromethane, 35 mL / min) to obtain 4-(((5'-chloro-2'-(((1R,4R)-4-((1-(1-(4-methoxybenzyl)-1H-tetrazole-5-yl)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 10, 150 mg, 207.83 umol, yield 18.17%, purity 93%) as a brown oil. MS (ESI) m / z = 671.2 [M+H] + .

[0398] Step 2: Preparation of 4-(((2'-(((1R,4R)-4-(((R)-1-(1H-tetrazole-5-yl)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 12) and 4-(((2'-(((1S,4R)-4-(((S)-1-(1H-tetrazole-5-yl)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 13)

[0399] [ka]

[0400] A solution of 4-(((5'-chloro-2'-(((1R,4R)-4-((1-(1-(4-methoxybenzyl)-1H-tetrazole-5-yl)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 10, 140 mg, 208.57 ml, 1 equivalent) in TFA (38.50 g, 337.65 mmol, 25.00 mL, 1618.88 equivalents) was stirred at 40°C for 12 hours, concentrated under vacuum, diluted with water (20 mL), and extracted with TBME (20 mL x 2). The aqueous layer was basicized to pH 8 with saturated NaHCO3 and extracted with DCM (40 mL x 5). The combined DCM layers were washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated to obtain 4-(((2'-(((1R,4R)-4-((1-(1H-tetrazole-5-yl)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (110 mg, crude) (compound 11) as a yellow oil. MS (ESI) m / z = 551.4 [M+H] + .

[0401] 4-(((2'-(((1R,4R)-4-((1-(1H-tetrazole-5-yl)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 11) is processed into chiral SFC (Chiralpak IC 250×30mm (particle size 10μm); 60% MeOH (0.1%) Further purification with NH3 / H2O / CO2) yielded 4-(((2'-(((1R,4R)-4-(((R)-1-(1H-tetrazole-5-yl)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 12, 30 mg, purity 76.7%) and 4-(((2'-(((1S,4R)-4-(((S)-1-(1H-tetrazole-5-yl)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 13, 50 mg, purity 93.6%). Compound 12 was further purified by preparative HPLC (Phenomenex Luna C18 80×40 mm (particle size 3 μm); 0-30% acetonitrile / water (0.05% HCl); 11 min) to obtain the hydrochloride salt of compound 12 (21 mg, 35.74 umol, yield 17.91%, purity >99%) as a yellow solid. Compound 13 was further purified by preparative HPLC (Phenomenex Luna C18 80×40 mm (particle size 3 μm); 0-30% acetonitrile / water (0.05% HCl); 11 min) to obtain the hydrochloride salt of compound 13 (26 mg, 44.25 umol, yield 22.17%, purity >99%) as a yellow solid.

[0402] Compound 12: 1H NMR (400 MHz, メタノール-d4) δ = 8.15 (s, 1H), 7.93 (t, J=8.1 Hz, 1H), 7.30 (s, 1H), 7.22 (d, J=8.8 Hz, 1H), 7.09 (d, J=7.2 Hz, 1H), 3.99 (dd, J=3.2, 11.2 Hz, 3H), 3.88 - 3.78 (m, 3H), 3.70 - 3.61 (m, 2H), 3.57 - 3.45 (m, 2H), 3.40 - 3.33 (m, 1H), 2.29 (t, J=14.4 Hz, 4H), 1.98 (d, J=13.6 Hz, 2H), 1.83 - 1.71 (m, 4H), 1.68 - 1.54 (m, 2H), 1.39 (d, J=6.4 Hz, 3H). MS (ESI) m / z = 551.2 [M+H] + .

[0403] Compound 13: 1 H NMR (400 MHz, メタノール-d4) δ = 8.17 (s, 1H), 8.03 (t, J=8.0 Hz, 1H), 7.35 (s, 2H), 7.13 (d, J=7.2 Hz, 1H), 4.00 (dd, J=3.2, 11.6 Hz, 3H), 3.89 (s, 2H), 3.87 - 3.79 (m, 1H), 3.70 - 3.61 (m, 2H), 3.57 - 3.47 (m, 2H), 3.41 - 3.33 (m, 1H), 2.30 (br t, J=14.4 Hz, 4H), 2.01 (br d, J=13.2 Hz, 2H), 1.85 - 1.73 (m, 4H), 1.69 - 1.56 (m, 2H), 1.40 (d, J=6.4 Hz, 3H). MS (ESI) m / z = 551.2 [M+H] + .

[0404] Example 9 Synthesis of イソプロピル2-(2-オキソプロポキシ)アセテート (Intermediate 5)

[0405]

change

[0406] Step 1: Preparation of isopropyl 2-((2-methylallyl)oxy)acetate (INT-17)

[0407] [ka]

[0408] To a solution of 2-methylpropa-2-en-1-ol (1.99 g, 27.62 mmol) in DMF (50 mL), NaH (1.66 g, 41.43 mmol, 60% purity) was added over 15 minutes at 0°C, followed by the slow addition of isopropyl 2-bromoacetate (INT-16, 5 g, 27.62 mmol). The mixture was stirred at 0°C for 1 hour. The mixture was quenched with water (25 mL) at 0°C and then extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated under vacuum, and purified by flash column chromatography on silica gel (0%-3% ethyl acetate in petroleum ether) to obtain isopropyl 2-(2-methylallyloxy)acetate (INT-17, 2.22 g, 46% yield) as a colorless liquid. 1 H NMR (400 MHz, DMSO-d6): δ = 5.05-4.8 (m, 3H), 4.03 (s, 2H), 4.0-3.85 (m, 2H), 1.68 (s, 3H), 1.21 (s, 3H), 1.19 (s, 3H).

[0409] Step 2: Preparation of ethyl 2-(2-oxopropoxy)acetate (intermediate 5)

[0410] [ka]

[0411] To a solution of isopropyl 2-(2-methylallyloxy)acetate (INT-17, 2.22 g, 12.89 mmol) in THF (25 mL) and H2O (24 mL), K2OsO4·2H2O (48 mg, 0.129 mmol) and NaIO4 (6.07 g, 28.36 mmol) were added. The mixture was stirred at 20°C for 12 hours. The mixture was diluted with H2O (10 mL) and extracted with DCM (20 mL x 3). The organic phases were combined, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to obtain a residue. This residue was purified by silica gel chromatography (10-15% ethyl acetate in petroleum ether) to obtain isopropyl 2-acetonyloxyacetate (intermediate 5, 1.06 g, yield 47%) as a pale yellow liquid. 1 H NMR (400 MHz, DMSO-d6): δ = 5.13-5.04 (m, 1H), 4.19 (s, 2H), 4.13 (s, 2H), 2.17 (s, 3H), 1.26 (s, 3H), 1.24 (s, 3H).

[0412] Example 10 Synthesis of isopropyl 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetate (compound 15) and isopropyl 2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetate (compound 16)

[0413] [ka]

[0414] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1, 100 mg, 0.227 mmol), isopropyl 2-acetonyl oxyacetate (intermediate 5, 43 mg, 0.249 mmol), and AcOH (41 mg, 0.680 mmol) were dissolved in DCE (2 mL), to which NaBH(OAc)3 (67 mg, 0.317 mmol) was added at 0°C. The mixture was stirred at 20°C for 20 hours. The mixture was quenched with H2O (5 mL). The resulting mixture was extracted with ELISA (20 mL x 3). The organic phases were combined, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, concentrated under vacuum, and purified by silica gel chromatography (15-20% MeOH in DCM) to obtain isopropyl 2-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]acetate (compound 14, 50 mg, yield 12%) as a dark brown solid.

[0415] Racemic isopropyl 2-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]acetate (compound 14, 50 mg, 0.083 mmol) is processed using chiral SFC (Chiralpak AD 250×30 mm (particle size 10 μm); 70%). The compounds were separated by EtOH / CO2 and further purified by preparative HPLC (HCOOH conditions) to obtain isopropyl 2-[(2R)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]acetate (compound 15, 3.9 mg, yield 8%) as a white solid, and isopropyl 2-[(2S)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]acetate (compound 16, 11.2 mg, yield 21%) as a brown solid.

[0416] Compound 15: 1 H NMR (400 MHz, CDCl3): δ = 8.49 (s, 1H), 8.09 (s, 1H), 7.52 (t, J = 7.6 Hz, 1H), 6.97 (d, J = 7.2 Hz, 1H), 6.6-6.5 (m, 2H), 5.15-5.0 (m, 1H), 4.86 (t, J = 6.8 Hz, 1H), 4.56 (s, 1H), 4.15-4.0 (m, 2H), 4.0-3.9 (m, 2H), 3.78 (d, J = 6.8 Hz, 2H), 3.75-3.7 (m, 2H), 3.7-3.5 (m, 3H). 3.4-3.25 (m, 1H), 3.05-2.9 (m, 1H), 2.3-2.1 (m, 4H), 1.95-1.85 (m, 2H), 1.8-1.7 (m, 2H), 1.65-1.45 (m, 2H), 1.3-1.2 (m, 11H).

[0417] Compound 16:1 H NMR (400 MHz, CDCl3): δ = 8.1 (s, 1H), 7.51-7.5 (m, 1H), 6.97 (d, J = 7.2 Hz, 1H), 6.51-6.5 (m, 2H), 5.2-5.0 (m, 1H), 4.82 (t, J = 7.6 Hz, 1H), 4.45 (d, J = 8.4 Hz, 1H), 4.05 (d, J = 0.8 Hz, 2H), 4.05-3.95 (m, 2H), 3.77 (d, J = 7.6 Hz, 2H), 3.75-3.65 (m, 2H), 3.6-3.45 (m, 2H), 3.4-3.3 (m, 1H), 3.15-3.0 (m, 1H), 2.65-2.5 (m, 1H), 2.2-2.1 (m, 2H), 2.1-1.95 (m, 2H), 1.95-1.85 (m, 2H), 1.8-1.75 (m, 2H), 1.3-1.25 (m, 8H), 1.25-1.15 (m, 2H), 1.06 (d, J = 6.4 Hz, 3H).

[0418] Example 11 Synthesis of tert-butyl 2-(2-oxopropoxy)acetate (intermediate 6)

[0419] [ka]

[0420] Step 1: Preparation of tert-butyl 2-((2-methylallyl)oxy)acetate (INT-19)

[0421] [ka]

[0422] To a solution of 2-methylpropa-2-en-1-ol (296 mg, 4.10 mmol) in toluene (18 mL), 18 mL of 50% NaOH aqueous solution was added, followed by NBu4HSO4 (1.13 g, 3.33 mmol). The mixture was stirred at 15-20°C for 30 minutes, then tert-butyl 2-bromoacetate (INT-18, 500 mg, 2.56 mmol) was slowly added, and the mixture was stirred at 15-20°C for 1.5 hours. The mixture was extracted with ethyl acetate (300 mL x 3). The combined organic phase was washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated under vacuum, and purified by silica gel chromatography (3% ethyl acetate in petroleum ether) to obtain tert-butyl 2-(2-methylallyloxy)acetate (INT-19, 0.35 g, yield 72%) as a colorless liquid. 1 H NMR (400 MHz, CDCl3): δ = 4.98 (s, 1H), 4.92 (s, 1H), 3.98 (s, 2H), 3.94 (s, 2H), 1.75 (s, 3H), 1.48 (s, 9H).

[0423] Step 2: Preparation of tert-butyl 2-(2-oxopropoxy)acetate (intermediate 6)

[0424] [ka]

[0425] To a solution of tert-butyl 2-(2-methylallyloxy)acetate (INT-19, 4.5 g, 24.16 mmol) in THF (50 mL) and H2O (50 mL), K2OsO4·2H2O (89 mg, 0.242 mmol) and NaIO4 (11.37 g, 53.16 mmol) were added. The mixture was stirred at 20°C for 12 hours. The mixture was diluted with H2O (100 mL) and extracted with DCM (200 mL x 3). The organic phases were combined, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, concentrated under vacuum, and purified by silica gel chromatography (6-10% ethyl acetate in petroleum ether) to obtain tert-butyl 2-acetonyloxyacetate (intermediate 6, 2.99 g, yield 66%) as a pale yellow liquid. 1 H NMR (400 MHz, CDCl3): δ = 4.17 (s, 2H), 4.05 (s, 2H), 2.20 (s, 3H), 1.46 (s, 9H).

[0426] Example 12 Synthesis of tert-butyl 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetate (compound 18) and tert-butyl 2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)acetate (compound 19)

[0427] [ka]

[0428] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1, 200 mg, 0.454 mmol), tert-butyl 2-acetonyl oxyacetate (94 mg, 0.499 mmol), and AcOH (82 mg, 1.36 mmol) were dissolved in DCE (2 mL), to which NaBH(OAc)3 (intermediate 6, 135 mg, 0.635 mmol) was added at 0°C. The mixture was stirred at 20°C for 20 hours. The mixture was quenched with H2O (5 mL). The resulting mixture was extracted with ELISA (20 mL x 3). The organic phases were combined, washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, concentrated under vacuum, and purified by silica gel chromatography (15-20% MeOH in DCM) to obtain tert-butyl 2-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]acetate (compound 17, 110 mg, yield 25%) as a dark brown solid.

[0429] Racemic tert-butyl 2-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]acetate (compound 17, 92 mg) is processed into chiral SFC (Chiralpak AD 250×30 mm (particle size 10 μm); 75% The compounds were separated by EtOH / CO2 and further purified by preparative HPLC (HCOOH conditions) to obtain tert-butyl 2-[(2R)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]acetate (compound 18, 20.3 mg, yield 22%) as a white solid, and tert-butyl 2-[(2S)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]acetate (compound 19, 26.2 mg, yield 28%) as a white solid.

[0430] Compound 18: 1 H NMR (400 MHz, CDCl3): δ = 8.47 (s, 1H), 8.09 (s, 1H), 7.52 (t, J = 7.2 Hz, 1H), 6.97 (d, J = 7.2 Hz, 1H), 6.6-6.5 (m, 2H), 5.0-4.8 (m, 1H), 4.58 (s, 1H), 4.1-3.9 (m, 4H), 3.8-3.75 (m, 2H), 3.75-3.65 (m, 4H), 3.65-3.55 (m, 1H). 3.4-3.3 (m, 1H), 3.05-2.95 (m, 1H), 2.3-2.1 (m, 4H), 1.97-1.87 (m 2H), 1.8-1.7 (m, 2H), 1.7-1.5 (m, 2H), 1.49 (s, 9H), 1.32-1.22 (m, 5H).

[0431] Compound 19: 1H NMR (400 MHz, CDCl3): δ = 8.1 (s, 1H), 7.51 (t,J = 8.0 Hz, 1H), 6.97 (d, J = 7.2 Hz, 1H), 6.6-6.4 (m, 2H), 4.81 (t, J = 6.4 Hz, 1H), 4.44 (d, J = 7.6 Hz, 1H), 4.1-4.0 (m, 4H), 3.77 (d, J = 7.2 Hz, 2H), 3.7 (t, J = 11.2 Hz, 2H), 3.6-3.45 (m, 2H), 3.4-3.3 (m, 1H), 3.15-3.0 (m, 1H), 2.7-2.45 (m, 1H), 2.2-2.1 (m, 2H), 2.1-1.95 (m, 2H), 1.95-1.9 (m, 2H), 1.8-1.75 (m, 2H), 1.48 (s, 9H), 1.3-1.25 (m, 2H), 1.25-1.2 (m, 2H), 1.06 (d, J = 6.4 Hz, 3H).

[0432] Example 13 Synthesis of ethyl (1-(5-(((2-methylallyl)oxy)methyl)-2H-tetrazole-2-yl)ethyl) carbonate (INT-21) and ethyl (1-(5-(((2-methylallyl)oxy)methyl)-1H-tetrazole-1-yl)ethyl) carbonate (INT-22)

[0433] [ka]

[0434] Step 1: Preparation of 5-(((2-methylallyl)oxy)methyl)-2H-tetrazole (INT-20)

[0435] [ka]

[0436] 5-(2-methylallyloxymethyl)-2-tetrahydropyran-2-yl-tetrazole (INT-9, 5.09 g, 21.36 mmol) was dissolved in MeOH (50 mL) and 4-methylbenzenesulfonic acid (3.68 g, 21.36 mmol) was added. The mixture was stirred at 20°C for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (0-50% ethyl acetate in petroleum ether) to obtain 5-(2-methylallyloxymethyl)-2H-tetrazole (INT-20, 600 mg, yield 18%) as a white solid. 1 H NMR (400 MHz, CDCl3) δ = 5.02 (d, J = 6.0 Hz, 2 H), 4.91 (s, 2 H), 4.07 (s, 2 H), 1.77 (s, 3 H).

[0437] Step 2: Preparation of ethyl (1-(5-(((2-methylallyl)oxy)methyl)-2H-tetrazole-2-yl)ethyl) carbonate (INT-21) and ethyl (1-(5-(((2-methylallyl)oxy)methyl)-1H-tetrazole-1-yl)ethyl) carbonate (INT-22)

[0438] [ka]

[0439] To a solution of 5-(2-methylallyloxymethyl)-2H-tetrazole (INT-20, 600 mg, 3.89 mmol) in DMF (6 mL), DIEA (5.03 g, 38.92 mmol) and 1-chloroethylethyl carbonate (5.94 g, 38.92 mmol) were added. The mixture was stirred at 70°C for 16 hours. The mixture was diluted with ethyl acetate (100 mL) and washed with brine (100 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue. This residue was purified by flash column chromatography (silica gel, 100-200 mesh, 0-10% ethyl acetate in petroleum ether) to obtain ethyl 1-[5-(2-methylallyloxymethyl)tetrazole-1-yl]ethyl carbonate (INT-21, 326 mg, yield 31%) as a yellow oil, and ethyl 1-[5-(2-methylalioxymethyl)tetrazole-2-yl]ethyl carbonate (INT-22, 464 mg, yield 44%) as a yellow oil.

[0440] INT-21: 1 H NMR (400 MHz, CDCl3): δ = 7.00 (q, J = 6.4 Hz, 1 H), 5.06-4.88 (m, 4 H), 4.254.14 (m, 2 H), 3.96 (q, J = 12.4 Hz, 2 H), 1.98 (d, J = 6.0 Hz, 3 H), 1.74 (s, 3 H), 1.31-1.27 (m, 3 H).

[0441] INT-22: 1 H NMR (400 MHz, CDCl3) δ = 7.19 (q, J = 6.4 Hz, 1 H), 5.00 (d, J = 27.2 Hz, 2 H), 4.76 (s, 2 H), 4.29-4.19 (m, 2 H), 4.05 (s, 2 H), 2.04-2.01 (m, 3 H), 1.77 (s, 3 H), 1.31-1.28 (m, 3 H).

[0442] Example 14 Synthesis of (R)-ethyl(1-(5-((2-oxopropoxy)methyl)-2H-tetrazole-2-yl)ethyl) carbonate (intermediate 7) and (S)-ethyl(1-(5-((2-oxopropoxy)methyl)-2H-tetrazole-2-yl)ethyl) carbonate (intermediate 8)

[0443] [ka]

[0444] To a solution of ethyl 1-[5-(2-methylallyloxymethyl)tetrazole-2-yl]ethyl carbonate (INT-21, 464 mg, 1.72 mmol) in THF (10 mL) and H2O (10 mL), K2OsO4·2H2O (32 mg, 0.086 mmol) and NaIO4 (845 mg, 3.95 mmol) were added. The mixture was stirred at 15°C for 4 hours. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic phases were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue. This residue was purified by flash column chromatography (silica gel, 100-200 mesh, 0-20% ethyl acetate in petroleum ether) to obtain 1-[5-(acetonyloxymethyl)tetrazole-2-yl]ethylethyl carbonate (INT-23, 400 mg, yield 86%) as a yellow oil.

[0445] 1-[5-(acetonyloxymethyl)tetrazol-2-yl]ethyl ethyl carbonate (INT-23, 400 mg, 1.47 mmol) was separated by chiral SFC to obtain [(1R)-1-[5-(acetonyloxymethyl)tetrazol-2-yl]ethyl]ethyl carbonate (intermediate 7, 143 mg, yield 36%, purity 90%) as a yellow oil, and [(1S)-1-[5-(acetonyloxymethyl)tetrazol-2-yl]ethyl]ethyl carbonate (intermediate 8, 290 mg, yield 73%) as a yellow oil.

[0446] Intermediate 7:1 H NMR (400 MHz, MeOD): δ = 7.23 (q, J = 6.0 Hz, 1 H), 4.87 (s, 2 H), 4.29 (s, 2 H), 4.26-4.18(m, 2 H), 2.12 (s, 3 H), 1.96 (d, J = 6.0 Hz, 3 H), 1.27 (t, J = 7.2 Hz, 3 H).

[0447] Intermediate 8: 1 H NMR (400 MHz, MeOD): δ = 7.23 (q, J = 6.0 Hz, 1 H), 4.87 (s, 2 H), 4.30 (s, 2 H), 4.25 - 4.17(m, 2 H), 2.12 (s, 3 H), 1.96 (d, J = 6.4 Hz, 3 H), 1.27 (t, J = 6.8 Hz, 3 H).

[0448] Example 15 Synthesis of (R)-ethyl(1-(5-((2-oxopropoxy)methyl)-1H-tetrazol-1-yl)ethyl) carbonate (intermediate 9) and (S)-ethyl(1-(5-((2-oxopropoxy)methyl)-1H-tetrazol-1-yl)ethyl) carbonate (intermediate 10)

[0449] [ka]

[0450] To a solution of ethyl 1-[5-(2-methylallyloxymethyl)tetrazole-1-yl]ethyl carbonate (INT-22, 326 mg, 1.21 mmol) in THF (7 mL) and H2O (7 mL), K2OsO4·2H2O (22 mg, 0.060 mmol) and NaIO4 (593 mg, 2.77 mmol) were added. The mixture was stirred at 15°C for 4 hours. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic phases were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue. This residue was purified by flash column chromatography (silica gel, 100-200 mesh, 0-25% ethyl acetate in petroleum ether), followed by chiral SFC to obtain [(1R)-1-[5-(acetonyloxymethyl)tetrazole-1-yl]ethyl]ethyl carbonate (intermediate 9, 142 mg, yield 43%) as a yellow oil, and [(1S)-1-[5-(acetonyloxymethyl)tetrazole-1-yl]ethyl]ethyl carbonate (intermediate 10, 195 mg, yield 59%) as a yellow oil.

[0451] Intermediate 9: 1 H NMR (400 MHz, MeOD): δ = 7.16 (q, J = 6.0 Hz, 1 H), 5.16-4.97 (m, 2 H), 4.35 (d, J = 2.8 Hz, 1 H), 4.22-4.14 (m, 2 H), 2.09 (s, 3 H), 1.95 (d, J = 6.0 Hz, 3 H), 1.25 (t, J = 6.8 Hz, 3 H).

[0452] Intermediate 10: 1 H NMR (400 MHz, MeOD): δ = 7.16 (q, J = 6.0 Hz, 1 H), 5.17-4.96 (m, 2 H), 4.35 (d, J = 2.8 Hz, 2 H), 4.23-4.12 (m, 2 H), 2.09 (s, 3 H), 1.95 (d, J = 6.0 Hz, 3 H), 1.25 (t, J = 7.2 Hz, 3 H).

[0453] Example 16 Synthesis of (R)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazol-2-yl)ethylethyl carbonate (compound 21) and (R)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazol-2-yl)ethylethyl carbonate (compound 22)

[0454] [ka]

[0455] To a solution of [(1R)-1-[5-(acetonyloxymethyl)tetrazole-2-yl]ethyl]ethyl carbonate (intermediate 7, 123 mg, 0.45 mmol) and 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1, 166 mg, 0.38 mmol) in DCE (2 mL), HOAc (68 mg, 1.13 mmol) and NaBH(OAc)3 (112 mg, 0.53 mmol) were added at 0°C. The mixture was stirred at 20°C for 16 hours. The mixture was quenched with water (50 mL) at 0°C and extracted with DCM (50 mL x 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue, which was purified by RP-HPLC (16-46% acetonitrile and 0.225% formic acid in water) to obtain [(1R)-1-[5-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxymethyl]tetrazole-2-yl]ethyl]ethyl carbonate (compound 20, 168 mg, yield 63%) as a yellow oil.

[0456] [(1R)-1-[5-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxymethyl]tetrazol-2-yl]ethyl]ethyl carbonate (compound 20, 100 mg, 0.14 mmol) was separated by chiral SFC to obtain [(1R)-1-[5-[[(2R)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl] Mino-cyclohexyl-amino-propoxy-methyl-tetrazol-2-yl-ethyl-ethyl carbonate (compound 21, 11.6 mg, yield 11%) was obtained as a white solid, and [(1R)-1-[5-[[(2S)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino-cyclohexyl-amino-propoxy-methyl-tetrazol-2-yl-ethyl-ethyl carbonate (compound 22, 22.4 mg, yield 20%) was obtained as a white solid.

[0457] Compound 21: 1 H NMR (400 MHz, MeOD): δ = 8.54 (s, 1 H), 7.97 (s, 1 H), 7.53 - 7.40 (m, 1 H), 7.25 (q, J = 6.0 Hz, 1 H), 6.84 (d, J = 7.2 Hz, 1 H), 6.70 (s, 1 H), 6.63 (d, J = 8.4 Hz, 1 H), 4.98 - 4.92 (m, 2 H), 4.28 - 4.13 (m, 2 H), 4.00 - 3.83 (m, 3 H), 3.75 (s, 2 H), 3.73 - 3.55 (m, 6 H), 2.25 - 2.11 (m, 4 H), 1.96 (d, J = 6.0 Hz, 3 H), 1.93 - 1.85 (m, 2 H), 1.82 - 1.71 (m, 2 H), 1.60 - 1.46 (m, 2 H), 1.38 - 1.21 (m, 8 H).

[0458] Compound 22: 1 H NMR (400 MHz, MeOD): δ = 8.55 (s, 1 H), 7.97 (s, 1 H), 7.53 - 7.45 (m, 1 H), 7.25 (q, J = 6.0 Hz, 1 H), 6.84 (d, J = 7.2 Hz, 1 H), 6.70 (s, 1 H), 6.62 (d, J = 8.0 Hz, 1 H), 4.87 (s, 2 H), 4.26 - 4.15 (m, 2 H), 3.99 - 3.92 (m, 2 H), 3.82 - 3.74 (m, 3 H), 3.69 - 3.59 (m, 4 H), 3.51 - 3.42 (m, 1H), 3.15 - 2.99 (m, 1 H), 2.21 - 2.07 (m, 4 H), 1.96 (d, J = 6.4 Hz, 3 H), 1.93 - 1.86 (m, 2 H), 1.82 - 1.72 (m, 2 H), 1.50 - 1.23 (m, 10 H).

[0459] Example 17 Synthesis of (S)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazol-2-yl)ethylethyl carbonate (compound 24) and (S)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazol-2-yl)ethylethyl carbonate (compound 25)

[0460] [ka]

[0461] To a solution of [(1S)-1-[5-(acetonyloxymethyl)tetrazole-2-yl]ethyl]ethyl carbonate (intermediate 8, 270 mg, 0.99 mmol) and 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1, 364 mg, 0.83 mmol) in DCE (3 mL), HOAc (149 mg, 2.48 mmol) and NaBH(OAc)3 (245 mg, 1.16 mmol) were added at 0°C. The mixture was stirred at 20°C for 16 hours. The mixture was quenched with water (50 mL) at 0°C and extracted with DCM (50 mL x 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue. This residue was purified by RP-HPLC (16-46% acetonitrile and 0.225% formic acid in water) to obtain [(1S)-1-[5-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxymethyl]tetrazole-2-yl]ethyl]ethyl carbonate (compound 23, 260 mg, yield 44%) as a yellow oil.

[0462] [(1S)-1-[5-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxymethyl]tetrazol-2-yl]ethyl]ethyl carbonate (compound 23, 100 mg, 0.14 mmol) was separated by chiral SFC to obtain [(1S)-1-[5-[[(2R)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl] Mino-cyclohexyl amino-propoxy-methyl tetrazole-2-yl ethyl ethyl carbonate (compound 24, 17.3 mg, yield 16%) was obtained as a white solid, and [(1S)-1-[5-[[(2S)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl amino-propoxy-methyl tetrazole-2-yl ethyl ethyl carbonate (compound 25, 28.5 mg, yield 26%) was obtained as a white solid.

[0463] Compound 24: 11H NMR (400 MHz, MeOD): δ = 8.53 (s, 1 H), 7.98 (s, 1 H), 7.53 - 7.45 (m, 1 H), 7.25 (q, J = 6.0 Hz, 1 H), 6.83 (d, J = 7.2 Hz, 1 H), 6.70 (s, 1 H), 6.63 (d, J = 8.4 Hz, 1 H), 4.92 (s, 2 H), 4.26 - 4.14 (m, 2 H), 3.99 - 3.86 (m, 3 H), 3.75 (s, 2 H), 3.72 - 3.60 (m, 5 H), 3.28 - 3.20 (m, 1 H), 2.25 - 2.12 (m, 4 H), 1.97 (d, J = 6.0 Hz, 3 H), 1.93 - 1.86 (m, 2 H), 1.83 - 1.72 (m, 2 H), 1.61 - 1.47 (m, 2 H), 1.40 - 1.31 (m, 5 H), 1.26 (t, J = 7.2 Hz, 3 H).

[0464] Compound 25: 1 1H NMR (400 MHz, MeOD): δ = 8.55 (s, 1 H), 7.97 (s, 1 H), 7.53 - 7.44 (m, 1 H), 7.25 (q, J = 6.4 Hz, 1 H), 6.84 (d, J = 6.8 Hz, 1 H), 6.69 (s, 1 H), 6.62 (d, J = 8.4 Hz, 1 H), 4.89 (s, 2 H), 4.26 - 4.14 (m, 2 H), 3.99 - 3.92 (m, 2 H), 3.82 - 3.74 (m, 3 H), 3.70 - 3.59 (m, 4 H), 3.52 - 3.42 (m, 1 H), 3.12 - 3.00 (m, 1 H), 2.20 - 2.07 (m, 4 H), 1.96 (d, J = 6.4 Hz, 3 H), 1.93 - 1.86 (m, 2 H), 1.82 - 1.71 (m, 2 H), 1.54 - 1.25 (m, 10 H)

[0465] Example 18 Synthesis of (R)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-1H-tetrazol-1-yl)ethylethyl carbonate (compound 27) and (R)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-1H-tetrazol-1-yl)ethylethyl carbonate (compound 28)

[0466] [ka]

[0467] To a solution of [(1R)-1-[5-(acetonyloxymethyl)tetrazol-1-yl]ethyl]ethyl carbonate (intermediate 9, 122 mg, 0.45 mmol) and 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1, 165 mg, 0.37 mmol) in DCE (2 mL), HOAc (67 mg, 1.12 mmol) and NaBH(OAc)3 (111 mg, 0.52 mmol) were added at 0°C. The mixture was stirred at 20°C under N2 for 16 hours. The mixture was quenched with water (50 mL) at 0°C and extracted with DCM (50 mL x 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue. This residue was purified by preparative HPLC (45-75% acetonitrile and 0.225% formic acid in water) to obtain [(1R)-1-[5-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxymethyl]tetrazole-1-yl]ethyl]ethyl carbonate (compound 26, 73 mg, yield 27%) as a yellow oil.

[0468] [(1R)-1-[5-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxymethyl]tetrazol-1-yl]ethyl]ethyl carbonate (compound 26, 73 mg, 0.11 mmol) was separated by chiral SFC to obtain [(1R)-1-[5-[[(2R)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl] Mino-cyclohexyl-amino-propoxy-methyl-tetrazol-1-yl-ethyl-ethyl carbonate (compound 27, 14.5 mg, yield 19%) was obtained as a white solid, and [(1R)-1-[5-[[(2S)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl-amino-propoxy-methyl-tetrazol-1-yl-ethyl-ethyl carbonate (compound 28, 17.1 mg, yield 21%) was obtained as a white solid.

[0469] Compound 27: 11H NMR (400 MHz, MeOD): δ = 7.95 (s, 1 H), 7.52 - 7.46 (m, 1 H), 7.02 (q, J = 6.0 Hz, 1 H), 6.85 (d, J = 6.8 Hz, 1 H), 6.69 (s, 1 H), 6.62 (d, J = 8.4 Hz, 1 H), 5.12 - 5.00 (m, 2 H), 4.26 - 4.14 (m, 2 H), 3.99 - 3.92 (m, 2 H), 3.75 (s, 2 H), 3.69 - 3.59 (m, 4 H), 3.51 - 3.45 (m, 1 H), 3.29 - 3.27 (m, 1 H), 3.18 - 3.12 (m, 1 H), 2.16 - 1.99 (m, 4 H), 1.95 (d, J = 6.0 Hz, 3 H), 1.92 - 1.90 (m, 1 H), 1.89 - 1.85 (m, 1 H), 1.82 - 1.72 (m, 2 H), 1.35 - 1.24 (m, 7 H), 1.16 - 1.06 (m, 3 H).

[0470] Compound 28: 1 1H NMR (400 MHz, MeOD): δ = 8.54 (s, 1 H), 7.97 (s, 1 H), 7.52 - 7.46 (m, 1 H), 6.93 (q, J = 6.0 Hz, 1 H), 6.84 (d, J = 7.2 Hz, 1 H), 6.70 (s, 1 H), 6.62 (d, J = 8.4 Hz, 1 H), 5.18 - 5.06 (m, 2 H), 4.25 - 4.13 (m, 2 H), 3.99 - 3.91 (m, 2 H), 3.86 - 3.77 (m, 1 H), 3.75 (s, 2 H), 3.73 - 3.46 (m, 5 H), 3.21 - 3.10 (m, 1 H), 2.25 - 2.08 (m, 4 H), 1.96 (d, J = 6.0 Hz, 3 H), 1.92 - 1.86 (m, 2 H), 1.82 - 1.71 (m, 2 H), 1.60 - 1.42 (m, 2 H), 1.38 - 1.24 (m, 8 H).

[0471] Example 19 Synthesis of (S)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-1H-tetrazol-1-yl)ethylethyl carbonate (compound 30) and (S)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-1H-tetrazol-1-yl)ethylethyl carbonate (compound 31)

[0472] [ka]

[0473] To a solution of [(1S)-1-[5-(acetonyloxymethyl)tetrazol-1-yl]ethyl]ethyl carbonate (intermediate 10, 175 mg, 0.64 mmol) and 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1, 236 mg, 0.54 mmol) in DCE (2 mL), HOAc (97 mg, 1.61 mmol) and NaBH(OAc)3 (159 mg, 0.75 mmol) were added at 0°C. The mixture was stirred at 20°C under N2 for 16 hours. The mixture was quenched with water (50 mL) at 0°C and extracted with DCM (50 mL x 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue, which was purified by RP-HPLC (15-45% acetonitrile and 0.225% formic acid in water) to obtain [(1S)-1-[5-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxymethyl]tetrazole-1-yl]ethyl]ethyl carbonate (compound 29, 204 mg, yield 54%) as a yellow oil.

[0474] [(1S)-1-[5-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxymethyl]tetrazol-1-yl]ethyl]ethyl carbonate (compound 29, 100 mg, 0.14 mmol) was separated by chiral SFC to obtain [(1S)-1-[5-[[(2R)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl] Mino-cyclohexyl-amino-propoxy-methyl-tetrazol-1-yl-ethyl-ethyl carbonate (compound 30, 25.4 mg, yield 25%) was obtained as a white solid, and [(1S)-1-[5-[[(2S)-2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl-amino-propoxy-methyl-tetrazol-1-yl-ethyl-ethyl carbonate (compound 31, 27.1 mg, yield 24%) was obtained as a white solid.

[0475] Compound 30: 1 H NMR (400 MHz, MeOD): δ = 7.95 (s, 1 H), 7.52 - 7.45 (m, 1 H), 7.06 - 6.98 (m, 1 H), 6.89 - 6.81 (m, 1 H), 6.69 (s, 1 H), 6.62 (d, J = 8.4 Hz, 1 H), 5.11 - 4.95 (m, 2 H), 4.26 - 4.14 (m, 2 H), 3.99 - 3.92 (m, 2 H), 3.75 (s, 2 H), 3.70 - 3.44 (m, 6 H), 3.16 - 3.10 (m, 1 H), 2.14 - 1.98 (m, 4 H), 1.95 (d, J = 6.4 Hz, 3 H), 1.92 - 1.85 (m, 2 H), 1.81 - 1.72 (m, 2 H), 1.34 - 1.21 (m, 7 H), 1.15 - 1.04 (m, 3 H).

[0476] Compound 31: 1 H NMR (400 MHz, MeOD): δ = 8.54 (s, 1 H), 7.97 (s, 1 H), 7.52 - 7.45 (m, 1 H), 6.94 (q, J = 6.4 Hz, 1 H), 6.84 (d, J = 6.8 Hz, 1 H), 6.69 (s, 1 H), 6.63 (d, J = 8.0 Hz, 1 H), 5.21 - 5.06 (m, 2 H), 4.28 - 4.12 (m, 2 H), 3.99 - 3.92 (m, 2 H), 3.88 - 3.83 (m, 1 H), 3.75 (s, 2 H), 3.70 - 3.52 (m, 5H), 3.26 - 3.12 (m, 1 H), 2.23 - 2.10 (m, 4 H), 1.96 (d, J = 6.0 Hz, 3 H), 1.92 - 1.85 (m, 2 H), 1.81 - 1.72 (m, 2 H), 1.59 - 1.44 (m, 2 H), 1.43 - 1.23 (m, 8 H).

[0477] Example 20 Synthesis of 2-methyl-5-(((2-methylallyl)oxy)methyl)-2H-tetrazole (INT-26) and 1-methyl-5-(((2-methylallyl)oxy)methyl)-1H-tetrazole (INT-27)

[0478] [ka]

[0479] Step 1: Preparation of 1-((2H-tetrazole-5-yl)methoxy)propan-2-one (INT-25)

[0480] [ka]

[0481] 5-(2-methylallyloxymethyl)-2-tetrahydropyran-2-yl-tetrazole (intermediate 2, 3.0 g, 12.59 mmol) was dissolved in MeOH (1 mL) and 4-methylbenzenesulfonic acid (2.2 g, 12.59 mmol) was added. The mixture was stirred at 20°C for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (silica gel, 100-200 mesh, 0-40% ethyl acetate in petroleum ether) to obtain 5-(2-methylallyloxymethyl)-2H-tetrazole (INT-25, 2.1 g, 13.62 mmol, yield 54%) as a colorless oil. 1 H NMR (400 MHz, CDCl3): δ 5.00 - 4.85 (m, 4 H), 3.95 (s, 2 H), 1.62 (s, 3 H).

[0482] Step 2: Preparation of 2-methyl-5-(((2-methylallyl)oxy)methyl)-2H-tetrazole (INT-26) and 1-methyl-5-(((2-methylallyl)oxy)methyl)-1H-tetrazole (INT-27)

[0483] [ka]

[0484] Diazomethyl(trimethyl)silane (2M, 9.73mL) was added to a solution of 5-(2-methylallyloxymethyl)-2H-tetrazole (INT-25, 1.0g, 6.49 mmol) in THF (10 mL). The mixture was stirred at 20°C for 2 hours. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel, 100-200 mesh, 10-30% ethyl acetate in petroleum ether) to obtain 2-methyl-5-(2-methylallyloxymethyl)tetrazole (INT-26, 470 mg, 2.79 mmol, yield 43%) as a colorless oil, and 1-methyl-5-(2-methylallyloxymethyl)tetrazole (INT-27, 330 mg, 1.96 mmol, yield 30%) as a colorless oil.

[0485] INT-26: 1 H NMR (400 MHz, CDCl3) δ 5.00 (d, J = 24.0 Hz, 2 H), 4.74 (s, 2 H), 4.36 (s, 3 H), 4.05 (s, 2 H), 1.77 (s, 3 H).

[0486] INT-27: 1 H NMR (400 MHz, CDCl3) δ 4.99 (d, J = 6.0 Hz, 2 H), 4.83 (s, 2 H), 4.14 (s, 3 H), 3.95 (s, 2 H), 1.75 (s, 3 H).

[0487] Example 21 Synthesis of 1-((2-methyl-2H-tetrazole-5-yl)methoxy)propan-2-one (intermediate 11)

[0488] [ka]

[0489] To a solution of 2-methyl-5-(2-methylallyloxymethyl)tetrazole (INT-26, 450 mg, 2.68 mmol) in THF (4 mL) and H2O (3 mL), K2OsO4·2H2O (49 mg, 0.13 mmol) and NaIO4 (1.32 g, 6.15 mmol) were added. The mixture was stirred at 20°C for 4 hours. The mixture was diluted with ethyl acetate (100 mL) and washed with brine (100 mL). The organic extract was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel, 100-200 mesh, 0-5% methanol in dichloromethane) to obtain 1-[(2-methyltetrazole-5-yl)methoxy]propan-2-one (intermediate 11, 310 mg, 1.82 mmol, yield 68%) as a colorless oil. 1H NMR (400 MHz, CDCl3): δ 4.87(s, 2 H), 4.37(s, 3 H), 4.22 (s, 2H), 2.18(s, 2 H).

[0490] Example 22 Synthesis of 1-((1-methyl-1H-tetrazole-5-yl)methoxy)propan-2-one (intermediate 12)

[0491] [ka]

[0492] To a solution of 1-methyl-5-(2-methylallyloxymethyl)tetrazole (INT-27, 310 mg, 1.84 mmol) in THF (3 mL) and H2O (2 mL), K2OsO4·2H2O (34 mg, 0.09 mmol) and NaIO4 (907 mg, 4.24 mmol) were added. The mixture was stirred at 20°C for 4 hours. The mixture was diluted with ethyl acetate (100 mL) and washed with brine (100 mL). The organic extract was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel, 100-200 mesh, 0-80% methanol in dichloromethane) to obtain 1-[(1-methyltetrazole-5-yl)methoxy]propan-2-one (intermediate 12, 210 mg, 1.23 mmol, yield 67%) as a black oil. 1 H NMR (400 MHz, CDCl3) δ 4.86 (s, 2 H), 4.16 (s, 3 H), 4.15 (s, 2 H), 2.05 (s, 2H).

[0493] Example 23 Synthesis of 4-(((5'-chloro-2'-(((1R,4r)-4-(((R)-1-((2-methyl-2H-tetrazole-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 33) and 4-(((5'-chloro-2'-(((1S,4r)-4-(((S)-1-((2-methyl-2H-tetrazole-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 34)

[0494] [ka]

[0495] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1, 600 mg, 1.36 mmol) and 1-[(2-methyltetrazole-5-yl)methoxy]propan-2-one (intermediate 11, 278 mg, 1.63 mmol) were dissolved in DCE (5 mL) and HOAc (245 mg, 4.08 mmol) and NaBH(OAc)3 (404 mg, 1.90 mmol) were added. The mixture was stirred at 20°C for 36 hours. The mixture was quenched with water (50 mL) at 0°C and extracted with DCM (50 mL x 3). The combined organic phases were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. Separation of residue by HPLC (column: Xtimate C18 150) * 40mm *The compound was purified by 10 μm; elution phase: [water (0.225% FA)-ACN]; B%: 10%~40%, 10 min) to obtain 4-[[[6-[5-chloro-2-[[4-[[1-methyl-2-[(2-methyltetrazole-5-yl)methoxyethyl]amino]cyclohexyl]amino]-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (compound 32, 600 mg, 736 μmol, yield 54%) as a yellow oil.

[0496] 4-(((5'-chloro-2'-(((1r,4r)-4-((1-((2-methyl-2H-tetrazole-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 32, 100 mg, 0.17 mmol) was separated by chiral SFC to obtain 4-(((5'-chloro-2'-(((1S,4r)-4-(((S)-1-((2-methyl-2H-tetrazole-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino (N)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 33, 11.2 mg, yield 10.5%, purity 94%) and (((5'-chloro-2'-(((1R,4r)-4-(((R)-1-((2-methyl-2H-tetrazole-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 34, 22 mg, yield 21.6%, purity 98%) were obtained as white solids.

[0497] Compound 33: 11H NMR (400 MHz, CDCl3): δ = 7.95 (s, 1 H), 7.50 - 7.46 (m, 1 H), 6.85 (d, J = 7.2 Hz, 1 H), 6.69 (s, 1 H), 6.62 (d, J = 8.4 Hz, 1 H), 4.76 (s, 2 H), 4.37 (s, 3 H), 3.97 - 3.93 (m, 2 H), 3.75 (s, 2 H), 3.66 - 3.55 (m, 4H), 3.45 - 3.41 (m, 1 H), 3.12 - 3.08 (m, 1 H), 2.64 - 2.63 (m, 1 H). 2.11 - 1.97 (m, 4 H), 1.90 - 1.87 (m, 2 H), 1.80 - 1.72 (m, 2 H), 1.32 - 1.22 (m, 4 H), 1.05 (d, J = 6.4 Hz, 3 H).

[0498] Compound 34: 1 1H NMR (400 MHz, CDCl3): δ = 8.54 (s, 1 H), 7.97 (s, 1 H), 7.51 - 7.47 (m, 1 H), 6.83 (d, J = 6.8 Hz, 1 H), 6.69 (s, 1 H), 6.62 (d, J = 8.4 Hz, 1 H), 4.89 (s, 2 H), 4.39 (s, 3 H), 3.97 - 3.94 (m, 2 H), 3.90 - 3.80 (m, 1 H), 3.75 (s, 2 H), 3.67 - 3.61 (m, 5 H), 3.23 - 3.17 (m, 1 H), 2.21 - 2.12 (m, 5 H), 1.91 - 1.87 (m, 2 H), 1.80 -​​​​​Synthesis of 4-(((5'-chloro-2'-(((1R,4r)-4-(((R)-1-((1-methyl-1H-tetrazole-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 36) and 4-(((5'-chloro-2'-(((1S,4r)-4-(((S)-1-((1-methyl-1H-tetrazole-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 37)

[0500] [ka]

[0501] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1, 410 mg, 0.93 mmol) and 1-[(1-methyltetrazole-5-yl)methoxy]propan-2-one (intermediate 12, 190 mg, 1.12 mmol) were dissolved in DCE (5 mL) and HOAc (168 mg, 2.79 mmol) and NaBH(OAc)3 (276 mg, 1.30 mmol) were added. The mixture was stirred at 20°C for 36 hours. The mixture was quenched with water (50 mL) at 0°C and extracted with DCM (50 mL x 3). The combined organic phases were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Separation of residue by HPLC (column: Xtimate C18 150) * 40mm *The compound was purified by 10 μm; elution phase: [water (0.225% FA)-ACN]; B%: 10%~40%, 10 min) to obtain 4-[[[6-[5-chloro-2-[[4-[[1-methyl-2-[(1-methyltetrazole-5-yl)methoxyethyl]amino]cyclohexyl]amino]-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (compound 35, 300 mg, 367.98 μmol, yield 40%) as a yellow oil.

[0502] 4-[[[6-[5-chloro-2-[[4-[[1-methyl-2-[(1-methyltetrazol-5-yl)methoxy]ethyl]amino]cyclohexyl]amino]-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (compound 35, 100 mg, 0.17 mmol) was separated by chiral SFC to obtain 4-[[[6-[5-chloro-2-[[4-[[(1S)-1-methyl-2-[(1-methyltetrazol-5-yl)methoxy]ethyl]amino]cyclohexyl]amino]- 4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (compound 36, 18.7 mg, yield 18.2%, purity 97%) and 4-[[[6-[5-chloro-2-[[4-[[(1R)-1-methyl-2-[(1-methyltetrazole-5-yl)methoxy]ethyl]amino]cyclohexyl]amino]-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (compound 37, 21.6 mg, yield 21.6%, purity 98%) were obtained as white solids.

[0503] Compound 36: 1H NMR (400 MHz, MeOD): δ = 7.95 (s, 1 H), 7.50 - 7.46 (m, 1H), 6.85 (d, J = 7.2 Hz, 1 H), 6.69 (s, 1 H), 6.62 (d, J = 8.4 Hz, 1 H), 4.90 (s, 2 H), 4.13 (s, 3 H), 3.97 - 3.93 (m, 2 H), 3.75 (s, 2 H), 3.66 - 3.61 (m, 3 H), 3.56 - 3.52 (m, 1 H), 3.48 - 3.44 (m, 1 H), 3.17 - 3.13 (m, 1 H). 2.66 (s, 1 H), 2.11 - 2.09 (m, 2H), 2.03 - 1.98 (m, 2 H), 1.90 - 1.87 (m, 2 H), 1.80 - 1.73 (m, 2 H), 1.31 - 1.24 (m, 4 H), 1.10 (d, J = 6.4 Hz, 3 H).

[0504] Compound 37: 1 H NMR (400 MHz, MeOD): δ = 7.96 (s, 1 H), 7.51 - 7.47 (m, 1 H), 6.84 (d, J = 7.2 Hz, 1 H), 6.69 (s, 1 H), 6.62 (d, J = 8.0 Hz, 1 H), 4.93 (s, 2 H), 4.13 (s, 3 H), 3.97 - 3.93 (m, 2 H), 3.75 (s, 2 H), 3.67 - 3.61 (m, 4 H), 3.56 - 3.52 (m, 1 H), 2.87 - 2.84 (m, 1H), 2.15 - 2.01 (m, 5 H), 1.90 - 1.87 (m, 2 H), 1.80 - 1.72 (m, 2 H), 1.39 - 1.29 (m, 4 H), 1.19 (d, J = 6.4 Hz, 3 H).

[0505] Example 25 Synthesis of tert-ブチル(2-(2-オキソプロポキシ)エチル)カルバメート(Intermediate 13)

[0506] [ka]

[0507] Step 1: Preparation of tert-butyl(2-(2-methylallyl)oxy)ethyl)carbamate (INT-29)

[0508] [ka]

[0509] To a solution of tert-butyl N-(2-hydroxyethyl)carbamate (INT-28, 1.0 g, 6.20 mmol) in DCM (12 mL), 3-bromo-2-methyl-propa-1-ene (921 mg, 6.82 mmol), TBAB (1.40 g, 4.34 mmol), and aqueous NaOH solution (10 M, 6 mL) were added. The mixture was stirred at 25°C for 12 hours. The mixture was diluted with water (10 mL) and extracted with DCM (20 mL x 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated under vacuum to obtain a residue, which was purified by silica gel chromatography (Biotage 4g Silica Flash Column; eluent of petroleum ether with a 0-10% gradient in ethyl acetate @ 40 mL / min) to obtain tert-butyl N-[2-(2-methylallyloxy)ethyl]carbamate (INT-29, 880 mg, yield 66%) as a colorless oil. 1 H NMR (400 MHz, CDCl3): δ = 4.96 (s, 1H), 4.91 (s, 1H), 3.89 (s, 2H), 3.49-3.46 (m, 2H), 3.35-3.32 (m, 2H), 1.74 (s, 3H), 1.45 (s, 9H).

[0510] Step 2: Preparation of tert-butyl(2-(2-oxopropoxy)ethyl)carbamate (intermediate 13)

[0511] [ka]

[0512] To a solution of tert-butyl N-[2-(2-methylallyloxy)ethyl]carbamate (200 mg, 0.93 mmol) in THF (3 mL) and H2O (3 mL), K2OsO4·2H2O (17 mg, 0.05 mmol) and NaIO4 (457 mg, 2.14 mmol) were added. The mixture was stirred at 15°C for 4 hours. The mixture was diluted with water (20 mL) and extracted with ethyl acetate (25 mL x 2). The combined organic phases were washed with an aqueous solution of Na2S2O3 (20 mL) and brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue. This residue was purified by silica gel chromatography (Biotage 4g Silica Flash Column; eluent of petroleum ether with a 0-50% gradient in ethyl acetate @ 40 mL / min) to obtain tert-butyl N-(2-acetonyloxyethyl)carbamate (intermediate 13, 200 mg, yield 99%) as a pale yellow oil. 1 H NMR (400 MHz, CDCl3): δ = 4.10 (s, 2H), 3.58-3.56 (m, 2H), 3.37-3.35 (m, 2H), 2.15 (s, 3H), 1.46 (s, 9H).

[0513] Example 26 Preparation of N-(2-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl)-1,1,1-trifluoromethanesulfonamide (compound 40)

[0514] [ka]

[0515] Step 1: Preparation of tert-butyl(2-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl)carbamate (compound 38)

[0516] [ka]

[0517] To a solution of 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1, 300 mg, 0.68 mmol) in DCE (5 mL), tert-butyl N-(2-acetonyloxyethyl)carbamate (intermediate 13, 163 mg, 0.75 mmol), HOAc (123 mg, 2.04 mmol), and NaBH(OAc)3 (202 mg, 0.95 mmol) were added at 0°C. The mixture was stirred under nitrogen at 20°C for 4 hours. The mixture was quenched with water (5 mL) at 0°C and then extracted with DCM (50 mL x 3). The combined organic phases were washed with saturated Na2CO3 aqueous solution (50 mL) and brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel chromatography (Biotage 4g Silica Flash Column; dichloromethane medium gradient 0-15% methanol eluent @ 40 mL / min) to obtain tert-butyl N-[2-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]ethyl]carbamate (compound 38, 250 mg, yield 58%) as a yellow oil.

[0518] Step 2: Preparation of 4-(((2'-(((1r,4r)-4-((1-(2-aminoethoxy)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 39)

[0519] [ka]

[0520] To a solution of tert-butyl N-[2-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]ethyl]carbamate (compound 38, 200 mg, 0.31 mmol) in DCM (5 mL), TFA (710.17 mg, 6.23 mmol) was added. The mixture was stirred at 20°C for 2 hours. After concentrating the mixture, it was extracted with DCM (30 mL x 3). The combined organic phases were washed with saturated Na2CO3 aqueous solution (30 mL) and brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 4-[[[6-[2-[[4-[[2-(2-aminoethoxy)-1-methyl-ethyl]amino]cyclohexyl]amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (compound 39, 210 mg, crude) as a yellow solid.

[0521] Step 3: Preparation of N-(2-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl)-1,1,1-trifluoromethanesulfonamide (compound 40)

[0522] [ka]

[0523] To a solution of 4-[[[6-[[[4-[[2-(2-aminoethoxy)-1-methyl-ethyl]amino]cyclohexyl]amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (compound 39, 165 mg, 0.30 mmol) in DCM (5 mL), TEA (77 mg, 0.76 mmol) and Tf2O (94 mg, 0.33 mmol) were added at -78°C. The mixture was stirred at 20°C for 3 hours. The mixture was quenched with water (5 mL) and extracted with DCM (30 mL x 3). The combined organic phases were washed with saturated Na2CO3 aqueous solution (30 mL) and brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The reaction product was concentrated and preparative TLC (dichloromethane:methanol = 10:1, R) was obtained. f The crude product was purified by preparative HPLC (instrument: PREP-HPLC-WI, column: Phenomenex Luna C18 100) and this was separated by preparative HPLC (instrument: PREP-HPLC-WI, column: Phenomenex Luna C18 100). * 30mm * Further purification by 3 μm; developing phase: water (0.225% FA)-ACN; B start: 6%, B end: 56%, gradient time (8 min)) yielded N-[2-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]ethyl]-1,1,1-trifluoromethanesulfonamide (compound 40, 83 mg, yield 60%, FA salt) as a pale yellow solid. 1H NMR (400 MHz, CDCl3): δ =8.37 (s, 1H), 8.04 (s, 1H), 7.51 (t, J = 7.6 Hz, 1H), 6.95 (d, J = 7.2 Hz, 1H), 6.59 (s, 1H), 6.52 (d, J = 8.4 Hz, 1H), 5.01-4.97 (m, 1H), 4.00-3.96 (m, 2H), 3.77-3.55 (m, 9H), 3.46 (brs, 3H), 3.08-3.02 (m, 1H), 2.28-2.13 (m, 4H), 1.90 (d, J) = 13.6 Hz, 2H), 1.80-1.63(m, 4H), 1.40 (d, J = 6.4 Hz, 3H), 1.35-1.26 (m, 2H).

[0524] Example 27 Synthesis of N-(2-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)ethyl)-1,1-difluoromethanesulfonamide (compound 41)

[0525] [ka]

[0526] To a solution of 4-[[[6-[[[4-[[2-(2-aminoethoxy)-1-methyl-ethyl]amino]cyclohexyl]amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (compound 39, 240 mg, 0.44 mmol) in DCM (3 mL), TEA (90 mg, 0.89 mmol) and difluoromethanesulfonyl chloride (67 mg, 0.44 mmol) were added at -78°C. The mixture was stirred at 20°C for 1 hour. The mixture was quenched with water (8 mL) and extracted with DCM (50 mL x 3). The combined organic phase was washed with saturated Na2CO3 (50 mL) and brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The reaction product was concentrated and preparative TLC (dichloromethane:methanol = 10:1, R f The crude product was purified by preparative HPLC (=0.13), and this was then separated by preparative HPLC (instrument: PREP-HPLC-WI, column: Phenomenex Luna C18 100). * 30mm * Further purification by 3 μm; developing phase: water (0.225% FA)-ACN; B start: 6%, B end: 56%, gradient time (8 min)) yielded N-[2-[2-[[4-[[5-chloro-4-[6-[(4-cyanotetrahydropyran-4-yl)methylamino]-2-pyridyl]-2-pyridyl]amino]cyclohexyl]amino]propoxy]ethyl]-1,1-difluoromethanesulfonamide (compound 41, 50.5 mg, yield 62%, FA salt) as a pale yellow solid. 1H NMR (400 MHz, CDCl3): δ =8.37 (s, 1H), 8.06 (s, 1H), 7.53 (t, J = 7.2 Hz, 1H), 6.97 (d, J = 7.6 Hz, 1H), 6.60 (s, 1H), 6.52 (d, J = 8.0 Hz, 1H), 6.20 (t, J = 54.0 Hz, 1H), 4.92-4.90 (m, 1H), 4.00-3.96 (m, 2H), 3.77-3.61 (m, 10H), 3.46 (brs, 3H), 3.08-3.02 (m, 1H), 2.29-2.21 (m, 4H), 1.90 (d, J = 13.6 Hz, 2H), 1.80-1.63(m, 4H), 1.41 (d, J = 6.8 Hz, 3H), 1.35-1.26 (m, 2H).

[0527] Example 28 Synthesis of 1-(2-oxopropoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (intermediate 14)

[0528] [ka]

[0529] Chloromethanesulfonyl chloride (INT-30) is converted to 1-(2-oxopropoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (intermediate 14) in five synthetic steps.

[0530] Example 29 Synthesis of 1-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (compound 43) and 1-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (compound 44)

[0531] [ka]

[0532] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) and 1-(2-oxopropoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (intermediate 14) are dissolved in DCE, to which HOAc and NaBH(OAc)3 are added. The mixture is stirred at 20°C for 36 hours. The mixture is quenched with water at 0°C and extracted with DCM. The combined organic phases are washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain 1-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (compound 42).

[0533] 1-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (compound 42) was separated by chiral SFC to obtain 1-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2 We obtain ,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (compound 43) and 1-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-N-(2,2,2-trifluoroethyl)methanesulfonamide (compound 44).

[0534] Example 30 Synthesis of ethyl 1-(2-oxopropoxy)cyclopropane-1-carboxylate (intermediate 15)

[0535] [ka]

[0536] Step 1: Preparation of ethyl 1-((2-methylallyl)oxy)cyclopropane-1-carboxylate (INT-32)

[0537] [ka]

[0538] NaH is added to a solution of ethyl 1-hydroxycyclopropane-1-carboxylate (INT-31) in DMF, and the reaction mixture is stirred at 0°C for 0.5 hours. 1-bromo-2-methyl-2-propene is added, and the reaction mixture is stirred at 25°C for 2 hours, quenched with brine, and extracted with ethyl 1-((2-methylallyl)oxy)cyclopropane-1-carboxylate (INT-32). The crude product is used directly in the next step without further purification.

[0539] Step 2: Preparation of ethyl 1-(2-oxopropoxy)cyclopropane-1-carboxylate (intermediate 15)

[0540] [ka]

[0541] To a solution of ethyl 1-((2-methylallyl)oxy)cyclopropane-1-carboxylate (INT-32) in THF and H2O, K2OsO4·2H2O and NaIO4 are added. The mixture is stirred at 15°C for 4 hours. The mixture is diluted with water and extracted with ethyl acetate. The combined organic phase is washed with aqueous Na2S2O3 solution and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue, which is purified by silica gel chromatography to obtain ethyl 1-(2-oxopropoxy)cyclopropane-1-carboxylate (intermediate 15).

[0542] Example 31 Synthesis of ethyl 1-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 46) and ethyl 1-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 47)

[0543] [ka]

[0544] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) and ethyl 1-(2-oxopropoxy)cyclopropane-1-carboxylate (intermediate 15) are dissolved in DCE, to which HOAc and NaBH(OAc)3 are added. The mixture is stirred at 20°C for 36 hours. The mixture is quenched with water at 0°C and extracted with DCM. The combined organic phases are washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain ethyl 1-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 45).

[0545] Ethyl 1-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 45) was separated by chiral SFC to obtain ethyl 1-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl) We obtain amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 46) and ethyl 1-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 47).

[0546] Example 32 Synthesis of ethyl 2-methyl-2-(2-oxopropoxy)propanoate (intermediate 16)

[0547] [ka]

[0548] Step 1: Preparation of ethyl 2-methyl-2-((2-methylallyl)oxy)propanoate (INT-34)

[0549] [ka]

[0550] NaH is added to a solution of ethyl 2-hydroxy-2-methylpropanoate (INT-33) in DMF, and the reaction mixture is stirred at 0°C for 0.5 hours. 1-bromo-2-methyl-2-propene is added, and the reaction mixture is stirred at 25°C for 2 hours, quenched with brine, and extracted with ethyl ethyl 2-methyl-2-((2-methylallyl)oxy)propanoate (INT-34). The crude product is used directly in the next step without further purification.

[0551] Step 2: Preparation of ethyl 2-methyl-2-(2-oxopropoxy)propanoate (intermediate 16)

[0552] [ka]

[0553] To a solution of ethyl 2-methyl-2-((2-methylallyl)oxy)propanoate (INT-34) in THF and H2O, K2OsO4·2H2O and NaIO4 are added. The mixture is stirred at 15°C for 4 hours. The mixture is diluted with water and extracted with ethyl acetate. The combined organic phase is washed with aqueous Na2S2O3 solution and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue, which is purified by silica gel chromatography to obtain ethyl 2-methyl-2-(2-oxopropoxy)propanoate (intermediate 16).

[0554] Example 33 Synthesis of ethyl 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropanoate (compound 49) and ethyl 2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropanoate (compound 50)

[0555] [ka]

[0556] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) and ethyl 2-methyl-2-(2-oxopropoxy)propanoate (intermediate 16) are dissolved in DCE, to which HOAc and NaBH(OAc)3 are added. The mixture is stirred at 20°C for 36 hours. The mixture is quenched with water at 0°C and extracted with DCM. The combined organic phases are washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain ethyl 2-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropanoate (compound 48).

[0557] Ethyl 2-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropanoate (compound 48) was separated by chiral SFC to obtain ethyl 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl We obtain ethyl(amino)-[2,4'-bipyridine]-2'-yl(amino)cyclohexyl(amino)propoxy)-2-methylpropanoate (compound 49) and ethyl(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl(amino)-[2,4'-bipyridine]-2'-yl(amino)cyclohexyl(amino)propoxy)-2-methylpropanoate (compound 50).

[0558] Example 34 Synthesis of ethyl(R)-2-(2-oxopropoxy)propanoate (intermediate 17)

[0559] [ka]

[0560] Step 1: Preparation of ethyl(R)-2-((2-methylallyl)oxy)propanoate (INT-36)

[0561] [ka]

[0562] NaH is added to a solution of ethyl(R)-2-hydroxypropanoate (INT-35) in DMF, and the reaction mixture is stirred at 0°C for 0.5 hours. 1-Bromo-2-methyl-2-propene is added, and the reaction mixture is stirred at 25°C for 2 hours, quenched with brine, and extracted with ethyl(R)-2-((2-methylallyl)oxy)propanoate (INT-36). The combined organic layer is washed with brine, dried over Na2SO4, filtered, and concentrated to obtain ethyl(R)-2-((2-methylallyl)oxy)propanoate (INT-36). The crude product is used directly in the next step without further purification.

[0563] Step 2: Preparation of ethyl(R)-2-(2-oxopropoxy)propanoate (intermediate 17)

[0564] [ka]

[0565] To a solution of ethyl(R)-2-((2-methylallyl)oxy)propanoate (INT-36) in THF and H2O, K2OsO4·2H2O and NaIO4 are added. The mixture is stirred at 15°C for 4 hours. The mixture is diluted with water and extracted with ethyl acetate. The combined organic phase is washed with aqueous Na2S2O3 solution and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue, which is purified by silica gel chromatography to obtain ethyl(R)-2-(2-oxopropoxy)propanoate (intermediate 17).

[0566] Example 35 Synthesis of ethyl(R)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 52) and ethyl(R)-2-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 53)

[0567] [ka]

[0568] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) and ethyl(R)-2-(2-oxopropoxy)propanoate (intermediate 17) are dissolved in DCE, to which HOAc and NaBH(OAc)3 are added. The mixture is stirred at 20°C for 36 hours. The mixture is quenched with water at 0°C and extracted with DCM. The combined organic phases are washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain ethyl(2R)-2-(2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 51).

[0569] Ethyl(2R)-2-(2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 51) was separated by chiral SFC to obtain ethyl(R)-2-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4 -yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 52) and ethyl(R)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 53) are obtained.

[0570] Example 36 Synthesis of ethyl(S)-2-(2-oxopropoxy)propanoate (intermediate 18)

[0571] [ka]

[0572] Step 1: Preparation of ethyl(S)-2-((2-methylallyl)oxy)propanoate (INT-38)

[0573] [ka]

[0574] NaH is added to a solution of ethyl(S)-2-hydroxypropanoate (INT-37) in DMF, and the reaction mixture is stirred at 0°C for 0.5 hours. 1-Bromo-2-methyl-2-propene is added, and the reaction mixture is stirred at 25°C for 2 hours, quenched with brine, and extracted with ethyl(S)-2-((2-methylallyl)oxy)propanoate (INT-38). The crude product is used directly in the next step without further purification.

[0575] Step 2: Preparation of ethyl(S)-2-(2-oxopropoxy)propanoate (intermediate 18)

[0576] [ka]

[0577] To a solution of ethyl(S)-2-((2-methylallyl)oxy)propanoate (INT-38) in THF and H2O, K2OsO4·2H2O and NaIO4 are added. The mixture is stirred at 15°C for 4 hours. The mixture is diluted with water and extracted with ethyl acetate. The combined organic phase is washed with aqueous Na2S2O3 solution and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue, which is purified by silica gel chromatography to obtain ethyl(S)-2-(2-oxopropoxy)propanoate (intermediate 18).

[0578] Example 37 Synthesis of ethyl(S)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 55) and ethyl(S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 56)

[0579] [ka]

[0580] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) and ethyl(S)-2-(2-oxopropoxy)propanoate (intermediate 18) are dissolved in DCE, to which HOAc and NaBH(OAc)3 are added. The mixture is stirred at 20°C for 36 hours. The mixture is quenched with water at 0°C and extracted with DCM. The combined organic phases are washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain ethyl(2S)-2-(2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 54).

[0581] Ethyl(2S)-2-(2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 54) was separated by chiral SFC to obtain ethyl(S)-2-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4 -yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 55) and ethyl(S)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 56) are obtained.

[0582] Example 38 Synthesis of 1-((6-nitropyridine-2-yl)methoxy)propan-2-one (intermediate 19)

[0583] [ka]

[0584] Step 1: Preparation of 2-(((2-methylallyl)oxy)methyl)-6-nitropyridine (INT-40)

[0585] [ka]

[0586] NaH is added to a solution of (6-nitropyridine-2-yl)methanol (INT-39) in DMF, and the reaction mixture is stirred at 0°C for 0.5 hours. 1-bromo-2-methyl-2-propene is added, and the reaction mixture is stirred at 25°C for 2 hours, quenched with brine, and extracted with ethyl acetate. The combined organic layers are washed with brine, dried over Na2SO4, filtered, and concentrated to obtain 2-(((2-methylallyl)oxy)methyl)-6-nitropyridine (INT-40). The crude product is used directly in the next step without further purification.

[0587] Step 2: Preparation of ethyl(S)-2-(2-oxopropoxy)propanoate (intermediate 19)

[0588] [ka]

[0589] To a solution of 2-(((2-methylallyl)oxy)methyl)-6-nitropyridine (INT-40) in THF and H2O, K2OsO4·2H2O and NaIO4 are added. The mixture is stirred at 15°C for 4 hours. The mixture is diluted with water and extracted with ethyl acetate. The combined organic phase is washed with aqueous Na2S2O3 solution and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue, which is purified by silica gel chromatography to obtain ethyl(S)-2-(2-oxopropoxy)propanoate (intermediate 19).

[0590] Example 39 Synthesis of N-(6-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)pyridine-2-yl)methanesulfonamide (compound 58) and N-(6-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)pyridine-2-yl)methanesulfonamide (compound 59)

[0591] [ka]

[0592] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) and ethyl(S)-2-(2-oxopropoxy)propanoate (intermediate 19) are dissolved in DCE, to which HOAc and NaBH(OAc)3 are added. The mixture is stirred at 20°C for 36 hours. The mixture is quenched with water at 0°C and extracted with DCM. The combined organic phases are washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC, reduced with iron and acetic acid, and mesylated with methanesulfonyl chloride and triethylamine to obtain N-(6-((2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)pyridine-2-yl)methanesulfonamide (compound 57).

[0593] N-(6-((2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)pyridine-2-yl)methanesulfonamide (compound 57) was separated by chiral SFC to obtain N-(6-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino We obtain N-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)pyridine-2-yl)methanesulfonamide (compound 58) and N-(6-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)pyridine-2-yl)methanesulfonamide (compound 59).

[0594] Example 40 Synthesis of ethyl(R)-3-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoate (compound 61) and ethyl(S)-3-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoate (compound 62)

[0595] [ka]

[0596] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) is treated with ethyl 2-butenoate to obtain ethyl 3-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoate (compound 60).

[0597] Ethyl 3-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoate (compound 60) was separated by chiral SFC to obtain ethyl(R)-3-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4 -yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoate (compound 61) and ethyl(S)-3-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoate (compound 62) are obtained.

[0598] Example 41 Synthesis of tert-butyl(R)-3-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoate (compound 64) and tert-butyl(S)-3-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoate (compound 65)

[0599] [ka]

[0600] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) is treated with tert-butyl 2-butenoate to obtain tert-butyl 3-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoate (compound 63).

[0601] tert-butyl3-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoate (compound 63) was separated by chiral SFC to obtain tert-butyl(R)-3-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran -4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoate (compound 64) and tert-butyl(S)-3-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoate (compound 65) are obtained.

[0602] Example 42 Synthesis of (R)-3-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoic acid (compound 66)

[0603] [ka]

[0604] tert-butyl(R)-3-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoate (compound 64) is treated with trifluoroacetic acid to obtain (R)-3-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoic acid (compound 66).

[0605] Example 43 Synthesis of (S)-3-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoic acid (compound 67)

[0606] [ka]

[0607] tert-butyl(S)-3-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoate (compound 65) is treated with trifluoroacetic acid to obtain (S)-3-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoic acid (compound 67).

[0608] Example 44 (R)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazole-2-yl)-2-methylpropyl pivalate (compound 68), (S)-1-(5 -(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazole-2-yl)-2-methylpropyl pivalate (compound 69), 4-(((5'-chloro-2'-(( Synthesis of (1R,4r)-4-(((R)-1-(((R)-1-((3,3-dimethylbuta-1-en-2-yl)oxy)-2-methylpropyl)-1H-tetrazol-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 70) and (S)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-1H-tetrazol-1-yl)-2-methylpropyl pivalate (compound 71)

[0609] [ka]

[0610] 4-(((2'-(((1R,4r)-4-(((R)-1-((2H-tetrazole-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 2) was treated with (S)-1-chloro-2-methylpropyl pivalate and Ag2O, and separated by chiral SFC to obtain (R)-1-(5-(((R)-2-(((1r,4 R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazole-2-yl)-2-methylpropyl pivalate (compound 68), (S)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4' -Bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazol-2-yl)-2-methylpropyl pivalate (compound 69), 4-(((5'-chloro-2'-(((1R,4r)-4-(((R)-1-((1-((R)-1-((3,3-dimethylbuta-1-en-2-yl)oxy)-2-methylpropyl)-1H-tetrazol-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'- We obtain bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 70) and (S)-1-(5-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-1H-tetrazole-1-yl)-2-methylpropyl pivalate (compound 71).

[0611] Example 45 (R)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazole-2-yl)-2-methylpropyl pivalate (compound 72), (S)-1-(5 -(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazole-2-yl)-2-methylpropyl pivalate (compound 73), 4-(((5'-chloro-2'-(( Synthesis of (1S,4r)-4-(((S)-1-((1-((R)-1-((3,3-dimethylbuta-1-en-2-yl)oxy)-2-methylpropyl)-1H-tetrazol-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 74) and (S)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-1H-tetrazol-1-yl)-2-methylpropyl pivalate (compound 75)

[0612] [ka]

[0613] 4-(((2'-(((1S,4r)-4-(((S)-1-((2H-tetrazole-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 3) was treated with (S)-1-chloro-2-methylpropyl pivalate and Ag2O, and separated by chiral SFC to obtain (R)-1-(5-(((S)-2-(((1r,4 S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazole-2-yl)-2-methylpropyl pivalate (compound 72), (S)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4' -Bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-2H-tetrazol-2-yl)-2-methylpropyl pivalate (compound 73), 4-(((5'-chloro-2'-(((1S,4r)-4-(((S)-1-((1-((R)-1-((3,3-dimethylbuta-1-en-2-yl)oxy)-2-methylpropyl)-1H-tetrazol-5-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'- We obtain bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 74) and (S)-1-(5-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)methyl)-1H-tetrazole-1-yl)-2-methylpropyl pivalate (compound 75).

[0614] Example 46 Synthesis of 1-((4-methoxypyrimidine-2-yl)methoxy)propan-2-one (intermediate 20)

[0615] [ka]

[0616] Step 1: Preparation of 4-methoxy-2-(((2-methylallyl)oxy)methyl)pyrimidine (INT-42)

[0617] [ka]

[0618] NaH is added to a solution of (4-methoxypyrimidine-2-yl)methanol (INT-41) in DMF, and the reaction mixture is stirred at 0°C for 0.5 hours. 1-bromo-2-methyl-2-propene is added, and the reaction mixture is stirred at 25°C for 2 hours, quenched with brine, and extracted with ethyl acetate. The combined organic layers are washed with brine, dried over Na2SO4, filtered, and concentrated to obtain 4-methoxy-2-(((2-methylallyl)oxy)methyl)pyrimidine (INT-42). The crude product is used directly in the next step without further purification.

[0619] Step 2: Preparation of 1-((4-methoxypyrimidine-2-yl)methoxy)propan-2-one (intermediate 20)

[0620] [ka]

[0621] To a solution of 4-methoxy-2-(((2-methylallyl)oxy)methyl)pyrimidine (INT-42) in THF and H2O, K2OsO4·2H2O and NaIO4 are added. The mixture is stirred at 15°C for 4 hours. The mixture is diluted with water and extracted with ethyl acetate. The combined organic phase is washed with aqueous Na2S2O3 and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue, which is purified by silica gel chromatography to obtain 1-((4-methoxypyrimidine-2-yl)methoxy)propan-2-one (intermediate 20).

[0622] Example 47 Synthesis of 4-(((5'-chloro-2'-(((1R,4r)-4-(((R)-1-((5-fluoro-6-oxo-1,6-dihydropyrimidine-2-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 77) and 4-(((5'-chloro-2'-(((1S,4r)-4-(((S)-1-((5-fluoro-6-oxo-1,6-dihydropyrimidine-2-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 78)

[0623] [ka]

[0624] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) and 1-((4-methoxypyrimidine-2-yl)methoxy)propan-2-one (intermediate 20) are dissolved in DCE, to which HOAc and NaBH(OAc)3 are added. The mixture is stirred at 20°C for 36 hours. The mixture is quenched with water at 0°C and extracted with DCM. The combined organic phases are washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC, silylated with TMSI, and fluorinated with SelectFluor to obtain 4-(((5'-chloro-2'-(((1r,4r)-4-((1-((5-fluoro-6-oxo-1,6-dihydropyrimidine-2-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 76).

[0625] 4-(((5'-chloro-2'-(((1r,4r)-4-((1-((5-fluoro-6-oxo-1,6-dihydropyrimidine-2-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 76) was separated by chiral SFC to obtain 4-(((5'-chloro-2'-(((1R,4r)-4-(((R)-1-((5-fluoro-6-oxo-1,6-dihydropyrimidine-2-yl)methoxy)propan-2-yl) We obtain pan-2-yl)amino)cyclohexyl)amino)[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 77) and 4-(((5'-chloro-2'-(((1S,4r)-4-(((S)-1-((5-fluoro-6-oxo-1,6-dihydropyrimidine-2-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 78).

[0626] Example 48 Synthesis of tert-butyl 1-(2-oxopropoxy)cyclopropane-1-carboxylate (intermediate 21)

[0627] [ka]

[0628] Step 1: Preparation of tert-butyl 1-((2-methylallyl)oxy)cyclopropane-1-carboxylate (INT-44)

[0629] [ka]

[0630] NaH is added to a solution of tert-butyl 1-hydroxycyclopropane-1-carboxylate (INT-43) in DMF, and the reaction mixture is stirred at 0°C for 0.5 hours. 1-bromo-2-methyl-2-propene is added, and the reaction mixture is stirred at 25°C for 2 hours, quenched with brine, and extracted with ethyl acetate. The combined organic layer is washed with brine, dried over Na2SO4, filtered, and concentrated to obtain tert-butyl 1-((2-methylallyl)oxy)cyclopropane-1-carboxylate (INT-44). The crude product is used directly in the next step without further purification.

[0631] Step 2: Preparation of tert-butyl 1-(2-oxopropoxy)cyclopropane-1-carboxylate (intermediate 21)

[0632] [ka]

[0633] To a solution of tert-butyl 1-((2-methylallyl)oxy)cyclopropane-1-carboxylate (INT-44) in THF and H2O, K2OsO4·2H2O and NaIO4 are added. The mixture is stirred at 15°C for 4 hours. The mixture is diluted with water and extracted with ethyl acetate. The combined organic phase is washed with aqueous Na2S2O3 and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue, which is purified by silica gel chromatography to obtain tert-butyl 1-(2-oxopropoxy)cyclopropane-1-carboxylate (intermediate 21).

[0634] Example 49 Synthesis of tert-butyl 1-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 80) and tert-butyl 1-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 81)

[0635] [ka]

[0636] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) and tert-butyl 1-(2-oxopropoxy)cyclopropane-1-carboxylate (intermediate 21) are dissolved in DCE, to which HOAc and NaBH(OAc)3 are added. The mixture is stirred at 20°C for 36 hours. The mixture is quenched with water at 0°C and extracted with DCM. The combined organic phases are washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain tert-butyl1-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 79).

[0637] tert-butyl 1-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 79) was separated by chiral SFC to obtain tert-butyl 1-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino) We obtain chloro(amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 80) and tert-butyl1-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 81).

[0638] Example 50 Synthesis of (R)-3-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoic acid (compound 66)

[0639] [ka]

[0640] tert-butyl 1-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 80) is treated with trifluoroacetic acid to obtain 1-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylic acid (compound 82).

[0641] Example 51 Synthesis of (S)-3-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)butanoic acid (compound 67)

[0642] [ka]

[0643] tert-butyl 1-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylate (compound 81) is treated with trifluoroacetic acid to obtain 1-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)cyclopropane-1-carboxylic acid (compound 83).

[0644] Example 52 Synthesis of tert-butyl 2-methyl-2-(2-oxopropoxy)propanoate (intermediate 22)

[0645] [ka]

[0646] Step 1: Preparation of tert-butyl 2-methyl-2-((2-methylallyl)oxy)propanoate (INT-46)

[0647] [ka]

[0648] NaH is added to a solution of tert-butyl 2-hydroxy-2-methylpropanoate (INT-45) in DMF, and the reaction mixture is stirred at 0°C for 0.5 hours. 1-bromo-2-methyl-2-propene is added, and the reaction mixture is stirred at 25°C for 2 hours, quenched with brine, and extracted with ethyl acetate. The combined organic layers are washed with brine, dried over Na2SO4, filtered, and concentrated to obtain tert-butyl 2-methyl-2-((2-methylallyl)oxy)propanoate (INT-46). The crude product is used directly in the next step without further purification.

[0649] Step 2: Preparation of tert-butyl 2-methyl-2-(2-oxopropoxy)propanoate (intermediate 22)

[0650] [ka]

[0651] To a solution of tert-butyl 2-methyl-2-((2-methylallyl)oxy)propanoate (INT-46) in THF and H2O, K2OsO4·2H2O and NaIO4 are added. The mixture is stirred at 15°C for 4 hours. The mixture is diluted with water and extracted with ethyl acetate. The combined organic phase is washed with aqueous Na2S2O3 solution and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue, which is purified by silica gel chromatography to obtain tert-butyl 2-methyl-2-(2-oxopropoxy)propanoate (intermediate 22).

[0652] Example 53 Synthesis of tert-butyl 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropanoate (compound 85) and tert-butyl 2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropanoate (compound 86)

[0653] [ka]

[0654] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) and tert-butyl 2-methyl-2-(2-oxopropoxy)propanoate (intermediate 22) are dissolved in DCE, to which HOAc and NaBH(OAc)3 are added. The mixture is stirred at 20°C for 36 hours. The mixture is quenched with water at 0°C and extracted with DCM. The combined organic phases are washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain tert-butyl 2-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropanoate (compound 84).

[0655] tert-butyl 2-(2-(((1r,4r)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropanoate (compound 84) was separated by chiral SFC to obtain tert-butyl 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl We obtain methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropanoate (compound 85) and tert-butyl 2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropanoate (compound 86).

[0656] Example 54 Synthesis of tert-butyl 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropanoic acid (compound 87)

[0657] [ka]

[0658] tert-butyl 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropanoate (compound 85) is treated with trifluoroacetic acid to obtain 2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropanoic acid (compound 87).

[0659] Example 55 Synthesis of tert-butyl 2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropanoic acid (compound 88)

[0660] [ka]

[0661] tert-butyl 2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropanoate (compound 86) is treated with trifluoroacetic acid to obtain 2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)-2-methylpropanoic acid (compound 88).

[0662] Example 56 Synthesis of tert-butyl(R)-2-(2-oxopropoxy)propanoate (intermediate 23)

[0663] [ka]

[0664] Step 1: Preparation of tert-butyl(R)-2-((2-methylallyl)oxy)propanoate (INT-48)

[0665] [ka]

[0666] NaH is added to a solution of tert-butyl(R)-2-hydroxypropanoate (INT-47) in DMF, and the reaction mixture is stirred at 0°C for 0.5 hours. 1-bromo-2-methyl-2-propene is added, and the reaction mixture is stirred at 25°C for 2 hours, quenched with brine, and extracted with ethyl acetate. The combined organic layer is washed with brine, dried over Na2SO4, filtered, and concentrated to obtain tert-butyl(R)-2-((2-methylallyl)oxy)propanoate (INT-48). The crude product is used directly in the next step without further purification.

[0667] Step 2: Preparation of tert-butyl(R)-2-(2-oxopropoxy)propanoate (intermediate 23)

[0668] [ka]

[0669] To a solution of tert-butyl(R)-2-((2-methylallyl)oxy)propanoate (INT-48) in THF and H2O, K2OsO4·2H2O and NaIO4 are added. The mixture is stirred at 15°C for 4 hours. The mixture is diluted with water and extracted with ethyl acetate. The combined organic phase is washed with aqueous Na2S2O3 solution and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue, which is purified by silica gel chromatography to obtain tert-butyl(R)-2-(2-oxopropoxy)propanoate (intermediate 23).

[0670] Example 57 Synthesis of tert-butyl(R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 90) and tert-butyl(R)-2-(((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 91)

[0671] [ka]

[0672] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) and tert-butyl(R)-2-(2-oxopropoxy)propanoate (intermediate 23) are dissolved in DCE, to which HOAc and NaBH(OAc)3 are added. The mixture is stirred at 20°C for 36 hours. The mixture is quenched with water at 0°C and extracted with DCM. The combined organic phases are washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain tert-butyl(2R)-2-(2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 89).

[0673] tert-butyl(2R)-2-(2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 89) was separated by chiral SFC to obtain tert-butyl(R)-2-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 89) and tert-butyl(R)-2-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 89) by chiral SFC. We obtain 4-yl(methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 90) and tert-butyl(R)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 91).

[0674] Example 58 Synthesis of (R)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoic acid (compound 92)

[0675] [ka]

[0676] tert-butyl(R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 90) is treated with trifluoroacetic acid to obtain (R)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoic acid (compound 92).

[0677] Example 59 Synthesis of (R)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoic acid (compound 93)

[0678] [ka]

[0679] tert-butyl(R)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 91) is treated with trifluoroacetic acid to obtain (R)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoic acid (compound 93).

[0680] Example 60 Synthesis of tert-butyl(S)-2-(2-oxopropoxy)propanoate (intermediate 24)

[0681] [ka]

[0682] Step 1: Preparation of tert-butyl(S)-2-((2-methylallyl)oxy)propanoate (INT-50)

[0683] [ka]

[0684] NaH is added to a solution of tert-butyl(S)-2-hydroxypropanoate (INT-49) in DMF, and the reaction mixture is stirred at 0°C for 0.5 hours. 1-bromo-2-methyl-2-propene is added, and the reaction mixture is stirred at 25°C for 2 hours, quenched with brine, and extracted with ethyl acetate. The combined organic layers are washed with brine, dried over Na2SO4, filtered, and concentrated to obtain tert-butyl(S)-2-((2-methylallyl)oxy)propanoate (INT-50). The crude product is used directly in the next step without further purification.

[0685] Step 2: Preparation of tert-butyl(S)-2-(2-oxopropoxy)propanoate (intermediate 24)

[0686] [ka]

[0687] To a solution of tert-butyl(S)-2-((2-methylallyl)oxy)propanoate (INT-50) in THF and H2O, K2OsO4·2H2O and NaIO4 are added. The mixture is stirred at 15°C for 4 hours. The mixture is diluted with water and extracted with ethyl acetate. The combined organic phase is washed with aqueous Na2S2O3 solution and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue, which is purified by silica gel chromatography to obtain tert-butyl(S)-2-(2-oxopropoxy)propanoate (intermediate 24).

[0688] Example 61 Synthesis of tert-butyl(S)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 95) and tert-butyl(S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 96)

[0689] [ka]

[0690] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) and tert-butyl(S)-2-(2-oxopropoxy)propanoate (intermediate 24) are dissolved in DCE, to which HOAc and NaBH(OAc)3 are added. The mixture is stirred at 20°C for 36 hours. The mixture is quenched with water at 0°C and extracted with DCM. The combined organic phases are washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain tert-butyl(2S)-2-(2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 94).

[0691] tert-butyl(2S)-2-(2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 94) was separated by chiral SFC to obtain tert-butyl(S)-2-(((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 94). We obtain 4-yl(methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 95) and tert-butyl(S)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 96).

[0692] Example 62 Synthesis of (S)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoic acid (compound 97)

[0693] [ka]

[0694] tert-butyl(S)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 95) is treated with trifluoroacetic acid to obtain (S)-2-((R)-2-(((1r,4R)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoic acid (compound 97).

[0695] Example 63 Synthesis of (S)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoic acid (compound 98)

[0696] [ka]

[0697] tert-butyl(S)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoate (compound 96) is treated with trifluoroacetic acid to obtain (S)-2-((S)-2-(((1r,4S)-4-((5'-chloro-6-(((4-cyanotetrahydro-2H-pyran-4-yl)methyl)amino)-[2,4'-bipyridine]-2'-yl)amino)cyclohexyl)amino)propoxy)propanoic acid (compound 98).

[0698] Example 64 Synthesis of 1-(1-(2-trityl-2H-tetrazole-5-yl)cyclopropoxy)propan-2-one (intermediate 25)

[0699] [ka]

[0700] Step 1: Preparation of 5-(1-((2-methylallyl)oxy)cyclopropyl)-2-trityl-2H-tetrazol (INT-52)

[0701] [ka]

[0702] NaH is added to a solution of 1-(2-trityl-2H-tetrazole-5-yl)cyclopropan-1-ol (INT-51) in DMF, and the reaction mixture is stirred at 0°C for 0.5 hours. 1-bromo-2-methyl-2-propene is added, and the reaction mixture is stirred at 25°C for 2 hours, quenched with brine, and extracted with ethyl acetate. The combined organic layers are washed with brine, dried over Na2SO4, filtered, and concentrated to obtain 5-(1-((2-methylallyl)oxy)cyclopropyl)-2-trityl-2H-tetrazole (INT-52). The crude product is used directly in the next step without further purification.

[0703] Step 2: Preparation of 1-(1-(2-trityl-2H-tetrazole-5-yl)cyclopropoxy)propan-2-one (intermediate 25)

[0704] [ka]

[0705] To a solution of 5-(1-((2-methylallyl)oxy)cyclopropyl)-2-trityl-2H-tetrazole (INT-52) in THF and H2O, K2OsO4·2H2O and NaIO4 are added. The mixture is stirred at 15°C for 4 hours. The mixture is diluted with water and extracted with ethyl acetate. The combined organic phase is washed with aqueous Na2S2O3 and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue, which is purified by silica gel chromatography to obtain 1-(1-(2-trityl-2H-tetrazole-5-yl)cyclopropoxy)propan-2-one (intermediate 25).

[0706] Example 65 Synthesis of 4-(((2'-(((1R,4r)-4-(((R)-1-(1-(2H-tetrazole-5-yl)cyclopropoxy)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 100) and 4-(((2'-(((1S,4r)-4-(((S)-1-(1-(2H-tetrazole-5-yl)cyclopropoxy)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 101)

[0707] [ka]

[0708] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) and 1-(1-(2-trityl-2H-tetrazole-5-yl)cyclopropoxy)propan-2-one (intermediate 25) are dissolved in DCE, to which HOAc and NaBH(OAc)3 are added. The mixture is stirred at 20°C for 36 hours. The mixture is quenched with water at 0°C and extracted with DCM. The combined organic phases are washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC and deprotected with HCl to obtain 4-(((2'-(((1r,4r)-4-((1-(1-(2H-tetrazole-5-yl)cyclopropoxy)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 99).

[0709] 4-(((2'-(((1r,4r)-4-((1-(1-(2H-tetrazole-5-yl)cyclopropoxy)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 99) was separated by chiral SFC to obtain 4-(((2'-(((1R,4r)-4-(((R)-1-(1-(2H-tetrazole-5-yl)cyclopropoxy)propane-2-yl)amino) We obtain chlorohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 100) and 4-(((2'-(((1S,4r)-4-(((S)-1-(1-(2H-tetrazole-5-yl)cyclopropoxy)propan-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 101).

[0710] Example 66 Synthesis of 1-((2-(2-trityl-2H-tetrazole-5-yl)propan-2-yl)oxy)propan-2-one (intermediate 26)

[0711] [ka]

[0712] Step 1: Preparation of 5-(2-((2-methylallyl)oxy)propan-2-yl)-2-trityl-2H-tetrazole (INT-54)

[0713] [ka]

[0714] NaH is added to a solution of 2-(2-trityl-2H-tetrazole-5-yl)propan-2-ol (INT-53) in DMF, and the reaction mixture is stirred at 0°C for 0.5 hours. 1-bromo-2-methyl-2-propene is added, and the reaction mixture is stirred at 25°C for 2 hours, quenched with brine, and extracted with siRNA. The combined organic layers are washed with brine, dried over Na2SO4, filtered, and concentrated to obtain 5-(2-((2-methylallyl)oxy)propan-2-yl)-2-trityl-2H-tetrazole (INT-54). The crude product is used directly in the next step without further purification.

[0715] Step 2: Preparation of 1-((2-(2-trityl-2H-tetrazole-5-yl)propan-2-yl)oxy)propan-2-one (intermediate 26)

[0716] [ka]

[0717] To a solution of 5-(2-((2-methylallyl)oxy)propan-2-yl)-2-trityl-2H-tetrazole (INT-54) in THF and H2O, K2OsO4·2H2O and NaIO4 are added. The mixture is stirred at 15°C for 4 hours. The mixture is diluted with water and extracted with ethyl acetate. The combined organic phase is washed with aqueous Na2S2O3 and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the residue, which is purified by silica gel chromatography to obtain 1-((2-(2-trityl-2H-tetrazole-5-yl)propan-2-yl)oxy)propan-2-one (intermediate 26).

[0718] Example 67 Synthesis of 4-(((2'-(((1R,4r)-4-(((R)-1-((2-(2H-tetrazole-5-yl)propane-2-yl)oxy)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 103) and 4-(((2'-(((1S,4r)-4-(((S)-1-((2-(2H-tetrazole-5-yl)propane-2-yl)oxy)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 104)

[0719] [ka]

[0720] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) and 1-((2-(2-trityl-2H-tetrazole-5-yl)propan-2-yl)oxy)propan-2-one (intermediate 26) are dissolved in DCE, to which HOAc and NaBH(OAc)3 are added. The mixture is stirred at 20°C for 36 hours. The mixture is quenched with water at 0°C and extracted with DCM. The combined organic phases are washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC and deprotected with HCl to obtain 4-(((2'-(((1r,4r)-4-((1-((2-(2H-tetrazole-5-yl)propane-2-yl)oxy)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 102).

[0721] 4-(((2'-(((1r,4r)-4-((1-((2-(2H-tetrazole-5-yl)propane-2-yl)oxy)propane-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 102) was separated by chiral SFC to obtain 4-(((2'-(((1R,4r)-4-(((R)-1-((2-(2H-tetrazole-5-yl)propane-2-yl)oxy)propane-2-yl)amino We obtain cyclohexyl(amino)-5'-chloro-[2,4'-bipyridine]-6-yl(amino)methyl(tetrahydro-2H-pyran-4-carbonitrile) (compound 103) and 4-(((2'-(((1S,4r)-4-(((S)-1-((2-(2H-tetrazole-5-yl)propan-2-yl)oxy(propan-2-yl)amino)cyclohexyl(amino)-5'-chloro-[2,4'-bipyridine]-6-yl(amino)methyl(tetrahydro-2H-pyran-4-carbonitrile) (compound 104).

[0722] Example 68 Synthesis of 1-(1-(2-trityl-2H-tetrazole-5-yl)ethoxy)propan-2-one (intermediate 27)

[0723] [ka]

[0724] Step 1: Preparation of 5-(1-((2-methylallyl)oxy)ethyl)-2-trityl-2H-tetrazole (INT-56)

[0725] [ka]

[0726] NaH is added to a solution of 1-(2-trityl-2H-tetrazole-5-yl)ethane-1-ol (INT-55) in DMF, and the reaction mixture is stirred at 0°C for 0.5 hours. 1-bromo-2-methyl-2-propene is added, and the reaction mixture is stirred at 25°C for 2 hours, quenched with brine, and extracted with ethyl acetate. The combined organic layers are washed with brine, dried over Na2SO4, filtered, and concentrated to obtain 5-(1-((2-methylallyl)oxy)ethyl)-2-trityl-2H-tetrazole (INT-56). The crude product is used directly in the next step without further purification.

[0727] Step 2: Preparation of 1-(1-(2-trityl-2H-tetrazole-5-yl)ethoxy)propan-2-one (intermediate 27)

[0728] [ka]

[0729] To a solution of 5-(1-((2-methylallyl)oxy)ethyl)-2-trityl-2H-tetrazole (INT-56) in THF and H2O, K2OsO4·2H2O and NaIO4 are added. The mixture is stirred at 15°C for 4 hours. The mixture is diluted with water and extracted with ethyl acetate. The combined organic phase is washed with aqueous Na2S2O3 and brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue, which is purified by silica gel chromatography to obtain 1-(1-(2-trityl-2H-tetrazole-5-yl)ethoxy)propan-2-one (intermediate 27).

[0730] Example 69 4-(((2'-(((1R,4r)-4-(((2R,3S)-3-((2H-tetrazole-5-yl)methoxy)butan-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 106), 4-(((2'-(((1S,4r)-4-(((S)-1-((R)-1-(2H-tetrazole-5-yl)ethoxy)propan-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 107), 4- Synthesis of (((2'-(((1R,4r)-4-(((2R,3R)-3-((2H-tetrazole-5-yl)methoxy)butan-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 108) and 4-(((2'-(((1S,4r)-4-(((S)-1-((S)-1-((S)-1-(2H-tetrazole-5-yl)ethoxy)propan-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 109)

[0731] [ka]

[0732] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) and 1-(1-(2-trityl-2H-tetrazole-5-yl)ethoxy)propan-2-one (intermediate 27) are dissolved in DCE, to which HOAc and NaBH(OAc)3 are added. The mixture is stirred at 20°C for 36 hours. The mixture is quenched with water at 0°C and extracted with DCM. The combined organic phases are washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC and deprotected with HCl to obtain 4-(((2'-(((1r,4r)-4-((1-((2-(2H-tetrazole-5-yl)propane-2-yl)oxy)propane-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 102).

[0733] 4-(((2'-(((1r,4r)-4-((1-((2-(2H-tetrazole-5-yl)propane-2-yl)oxy)propane-2-yl)amino)cyclohexyl)amino)5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 102) was separated by chiral SFC to obtain 4-(((2'-(((1R,4r)-4-(((2 R,3S)-3-((2H-tetrazol-5-yl)methoxy)butan-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 106), 4-(((2'-(((1S,4r)-4-(((S)-1-((R)-1-(2H-tetrazol-5-yl)ethoxy)propan-2-yl)amino (no)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 107), 4-(((2'-(((1R,4r)-4-(((2R,3R)-3-((2H-tetrazole-5-yl)methoxy)butan-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino) We obtain methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 108) and 4-(((2'-(((1S,4r)-4-(((S)-1-((S)-1-(2H-tetrazole-5-yl)ethoxy)propan-2-yl)amino)cyclohexyl)amino)-5'-chloro-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 109).

[0734] Example 70 Synthesis of 5-((2-oxopropoxy)methyl)-2,4-dihydro-3H-1,2,4-triazole-3-one (intermediate 28)

[0735] [ka]

[0736] Step 1: Preparation of 5-(hydroxymethyl)-2,4-dihydro-3H-1,2,4-triazole-3-one (INT-58)

[0737] [ka]

[0738] Hydrazine carboxamide hydrochloride (INT-57) is converted to 5-(hydroxymethyl)-2,4-dihydro-3H-1,2,4-triazole-3-one (INT-58) in three synthetic steps.

[0739] Step 2: Preparation of 5-((2-oxopropoxy)methyl)-2,4-dihydro-3H-1,2,4-triazole-3-one (intermediate 28)

[0740] [ka]

[0741] 5-hydroxymethyl)-2,4-dihydro-3H-1,2,4-triazole-3-one (INT-58) is converted to 5-((2-oxopropoxy)methyl)-2,4-dihydro-3H-1,2,4-triazole-3-one (intermediate 28) in four synthetic steps.

[0742] Example 71 4-(((5'-chloro-2'-(((1R,4r)-4-(((R)-1-((5-oxo-4,5-dihydro-1H-1,2,4-triazole-3-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 111) and Synthesis of 4-(((5'-chloro-2'-(((1S,4r)-4-(((S)-1-((5-oxo-4,5-dihydro-1H-1,2,4-triazole-3-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 112)

[0743] [ka]

[0744] 4-[[[6-[2-[(4-aminocyclohexyl)amino]-5-chloro-4-pyridyl]-2-pyridyl]amino]methyl]tetrahydropyran-4-carbonitrile (intermediate 1) and 5-((2-oxopropoxy)methyl)-2,4-dihydro-3H-1,2,4-triazole-3-one (intermediate 28) are dissolved in DCE, to which HOAc and NaBH(OAc)3 are added. The mixture is stirred at 20°C for 36 hours. The mixture is quenched with water at 0°C and extracted with DCM. The combined organic phases are washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to obtain 4-(((5'-chloro-2'-(((1r,4r)-4-((1-((5-oxo-4,5-dihydro-1H-1,2,4-triazole-3-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 110).

[0745] 4-(((5'-chloro-2'-(((1r,4r)-4-((1-((5-oxo-4,5-dihydro-1H-1,2,4-triazole-3-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 110) was separated by chiral SFC to obtain 4-(((5'-chloro-2'-(((1R,4r)-4-(((R)-1-((5-oxo-4,5-dihydro-1H-1,2,4-triazole-3-yl)methoxy) Propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 111) and 4-(((5'-chloro-2'-(((1S,4r)-4-(((S)-1-((5-oxo-4,5-dihydro-1H-1,2,4-triazole-3-yl)methoxy)propan-2-yl)amino)cyclohexyl)amino)-[2,4'-bipyridine]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile (compound 112) are obtained.

[0746] Example 72 CDK9 Biochemical Assay Each well of a 96-well plate was filled with 6 μL of 5x kinase assay buffer containing 10 mM DTT, 1 μL of 500 μM ATP, 10 μL of 5x CDK substrate, and 8 μL of water. 5 μL of the compound was added to the test group and positive control group, while 5 μL of the solvent was added to the blank group. 100 ng of CDK9 / CyclinT dissolved in 20 μL of water was added to the test group and positive control group, while 20 μL of 1x kinase assay buffer was added to the blank group. The reaction mixture was incubated at 30°C for 45 minutes, 50 μL of Kinase-Glo® Max was added to each well, and the plate was incubated at room temperature for 15 minutes in the dark. Luminescence was measured using a microplate reader, and IC50 was measured. 50 The values ​​were calculated using Prism 9 software.

[0747] IC obtained according to the above procedure 50 The values ​​are summarized in Table 1.

[0748] [Table 1]

[0749] Example 73 Cancer cell survival assay Human liver cancer cell lines HepG2, Hep3B, Huh7, and SK-HEP-1 cells were washed and trypsinized with 0.25% trypsin (Corning #25-053-CI) in a 37°C incubator until detached. These cells were resuspended and seeded in 96-well plates at a density of 5,000 cells / well. After cell adhesion, the compound was added at its final concentration, a 2-fold dilution. 72 hours after treatment with the compound, CellTiter-Glo® 2.0 was added to the wells in a medium / CellTiter-Glo® 2.0 ratio of 2:1. The plate was shielded from light, shaken for 2 minutes, and incubated for 10 minutes. Luminescence was measured using a microplate reader, and EC was measured. 50 The values ​​were calculated using Prism 9 software.

[0750] EC obtained according to the above procedure 50 The values ​​are summarized in Table 2.

[0751] [Table 2]

[0752] Example 74 Pharmacokinetic and tissue distribution studies. In mouse experiments, CD-1 mice were given a single oral dose of a 5 mg / kg suspension of the compounds, and the pharmacokinetics of the compounds in the liver and blood were analyzed. Liver and blood samples for determining compound concentrations were obtained 2 hours after compound administration (n=3 for each compound). The ratios of compound concentrations in the liver and blood of CD-1 mice after a single oral dose of a 5 mg / kg suspension of the compounds are summarized in Figure 1. The data in Figure 1 show that compounds 18, 40, and 33 had higher liver / blood ratios than NVP-2, demonstrating that these compounds possess improved liver selectivity.

[0753] In rat experiments, Sprague-Dawley (SD) rats were given a single oral dose of a 5 mg / kg suspension of the compounds, and the pharmacokinetics of the compounds in blood collected from the jugular and portal veins were analyzed. Blood samples for determining compound concentrations were obtained from the jugular and portal veins at 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, and 24 hours after compound administration (n=3 for each compound). The collected liver and blood samples were analyzed using LC-MS / MS to quantify the compound concentrations. The ratios of compound concentrations in blood collected from the jugular and portal veins of Sprague-Dawley (SD) rats after a single oral dose of a 5 mg / kg suspension of the compounds are summarized in Figure 2. The data in Figure 2 show that compounds 6 and 18 have lower hepatic outflow / inflow ratios than NVP-2, demonstrating that these compounds have improved hepatic selectivity.

[0754] Example 75 Tolerance and safety evaluation tests BALB / c nude mice were randomly assigned to each group based on body weight using a computer-based randomization procedure. Body weight of all animals was measured daily, and the change in body weight (relative to day 1) of BALB / c nude mice treated with the vehicle and compound was recorded. Routine monitoring included monitoring all test animals for behavior such as mobility, food and water consumption, body weight, loss of eye / coat luster, and any other abnormal effects. All mortality and / or abnormal clinical signs were recorded. Animals were euthanized when they experienced significant weight loss (emaciated, obvious weight loss of more than 20%). The mean change in body weight (relative to day 1) of BALB / c nude mice treated with the vehicle and compound is summarized in Figure 3. The data in Figure 3 demonstrate that compound 18 is less toxic than NVP-2.

[0755] Preferred embodiments of the present invention have been shown and described herein, but it will be apparent to those skilled in the art that such embodiments are provided merely as examples. Numerous variations, modifications, and substitutions will now be conceivable to those skilled in the art without departing from the present invention. It should be understood that various alternatives to the embodiments of the present invention described herein may be adopted in practicing the present invention. The following claims define the scope of the present invention, and methods and structures falling within these claims, as well as their equivalents, are intended to be covered thereby.

Claims

【Request Item 1】 【Chemistry 1-1】 [Chemistry 1-2] [Chemistry 1-3] [Chemistry 1-4] [Chemistry 1-5] A compound, or a pharmaceutically acceptable salt thereof, selected from the group consisting of the following. 【Request Item 2】 【Chemistry 2】 A compound, or a pharmaceutically acceptable salt thereof, selected from the group consisting of the following. 【Request Item 3】 【Chemistry 3】 The compound according to claim 2, or a pharmaceutically acceptable salt thereof. 【Request Item 4】 【Chemistry 4】 The compound according to claim 2, or a pharmaceutically acceptable salt thereof. 【Request Item 5】 【Chemistry 5】 The compound according to claim 2, or a pharmaceutically acceptable salt thereof. 【Request Item 6】 【Transformation 6】 The compound according to claim 2, or a pharmaceutically acceptable salt thereof.

7. A pharmaceutical composition comprising a compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

8. Use of a compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, in the manufacture of a drug for treating cancer in a patient requiring cancer treatment.

9. The use according to claim 8, wherein the cancer is selected from leukemia, breast cancer, prostate cancer, ovarian cancer, colon cancer, cervical cancer, lung cancer, lymphoma, and liver cancer.

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