DNA-dependent protein kinase inhibitor

Abstract

Disclosed herein are compounds of formula (I), and pharmaceutically acceptable salts thereof, that are useful as DNA-PK inhibitors. Also disclosed are pharmaceutical compositions comprising one or more compounds of formula (I), and methods of treating DNA-PK-related diseases (e.g., cancer) using such compounds or compositions.

Description

Technical Field

[0001] This specification generally relates to novel compounds that selectively modulate DNA-dependent protein kinase (“DNA-PK”), and pharmaceutically acceptable salts thereof. The disclosure also relates to pharmaceutical compositions comprising one or more compounds as active ingredients, and the use of the compounds in the treatment of DNA-PK related diseases including cancer.

Background Art

[0002] DNA-PK is a nuclear serine / threonine protein kinase complex composed of the catalytic subunit DNA-PKcs and the heterodimer of Ku proteins (Ku70 / Ku80). Functionally, DNA-PK is an essential component in the repair of DNA double-strand breaks (DSBs), plays a role in maintaining genomic integrity and the process of V(D)J recombination, and gives rise to a highly diverse repertoire of antibodies / immunoglobulins and T cell receptors found on B and T cells, respectively. In addition, DNA-PK and its components are involved in various other physiological processes including chromatin structure regulation, telomere maintenance, transcriptional control and response to replication stress (Smith and Jackson, 1999; Goodwin and Knudsen, 2014).

[0003] The human genome in the form of DNA is constantly exposed to the attack of reactive oxygen species (ROS), which are mainly by-products of oxidative metabolism. ROS can cause DNA damage in the form of single-strand breaks. DSBs may occur when single-strand breaks occur in close proximity previously. In addition, single-strand and single-strand-double-strand breaks are caused when DNA replication forks encounter damaged base patterns. Also, external influences such as ionizing radiation (e.g., gamma or particle radiation) and certain anti-cancer drugs (e.g., B. bleomycin) can induce DSBs. DSBs also occur as intermediates of somatic recombination, an important process in the formation of a functional immune system in all vertebrates.

[0004] If the DSBs are not repaired or are inaccurately repaired, mutations and / or chromosomal abnormalities that can lead to cell death occur. To cope with the severe threat imposed by DSBs, eukaryotic cells have evolved several mechanisms (e.g., DNA non-homologous end joining (NHEJ) and homologous recombination (HR)) to mediate these repairs, and DNA-PK plays an important role in these. Biochemical assays have shown that DNA-PK is most efficiently activated by the appearance of DNA DSBs. Cell lines with mutated and non-functional DNA-PK components have been found to be sensitive to radiation (Smith and Jackson, 1999). DNA-PK inhibitors may be effective as single agents in tumors with high endogenous levels of DNA damage. DNA-PK inhibitors are useful in oncology and, as either single-agent therapy or in combination with other agents, have been shown to include targeting tumors with high levels of replication stress (Lin et al., 2014; Ashley et al., 2014; Buisson et al., 2015) in prostate cancer (Goodwin et al., 2013) and breast cancer (Medunjanin et al., 2010).

[0005] Therefore, compounds that inhibit DNA-PK are required as pharmacological means and are important for the purpose of drugs for treating DNA-PK related diseases, such as cancer.

Summary of the Invention

[0006] In one aspect, the present disclosure provides a compound of formula (I):

[0007]

Chemical formula

[0008] or a pharmaceutically acceptable salt thereof (wherein X1, X2, X3, R 1 , R 2 , R 3 , R 4and ring A are as defined herein) provides.

[0009] In another aspect, the present disclosure provides a compound of formula (Ia):

[0010]

Chemical formula

[0011] or a pharmaceutically acceptable salt thereof (wherein R 1 , R 2 , R 3 , R 4 , n and ring A are as defined herein) provides.

[0012] In another aspect, the present disclosure provides a compound of formula (Ib):

[0013]

Chemical formula

[0014] or a pharmaceutically acceptable salt thereof (wherein R 1 , R 2 , R 3 , R 4 and ring A are as defined herein) provides.

[0015] In another aspect, the present disclosure provides a compound of formula (Ic):

[0016]

Chemical formula

[0017] or a pharmaceutically acceptable salt thereof (wherein R 1 , R 2 , R 3 , R4 and ring A are as defined herein) To provide.

[0018] In another aspect, the present disclosure provides a compound of formula (Id):

[0019]

Chemical formula

[0020] or a pharmaceutically acceptable salt thereof (wherein R 1 , R 2 , R 3 , R 4 and ring A are as defined herein) To provide.

[0021] In another aspect, the present disclosure provides a compound of formula (Ie):

[0022]

Chemical formula

[0023] and a pharmaceutically acceptable salt thereof (wherein X1, X3, Y1, Y2, Y3, R 1 , R 2 , R 5 and R 6 are as defined herein) To provide.

[0024] In yet another aspect, the present disclosure provides a pharmaceutical composition comprising, as an active ingredient, one or more compounds of formula (I), formula (Ia), formula (Ib), formula (Ic), formula (Id), formula (Ie), or a pharmaceutically acceptable salt thereof.

[0025] In another aspect, the present disclosure further provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, or one or more pharmaceutical compositions thereof, which are used to inhibit DNA-PK kinase.

[0026] In yet another aspect, the present disclosure provides the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, or one or more pharmaceutical compositions thereof, in the manufacture of a medicament for inhibiting DNA-PK kinase in a subject.

[0027] In yet another aspect, the present disclosure provides a method for inhibiting DNA-PK kinase by using one or more compounds of formula (I), or a pharmaceutically acceptable salt thereof, or one or more pharmaceutical compositions thereof.

[0028] In another aspect, the present disclosure provides a method for treating a DNA-PK related disease (e.g., cancer) by using a compound of formula (I), or a pharmaceutically acceptable salt thereof, or one or more pharmaceutical compositions thereof. In a further aspect, the present disclosure provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, in combination with a second therapeutic agent, preferably an anti-tumor agent.

[0029] In yet another aspect, the present disclosure provides a combination use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, and a second therapeutic agent, preferably an anti-tumor agent.

BRIEF DESCRIPTION OF THE INVENTION

[0030] Compound In one aspect, the present disclosure is directed to formula (I)

[0031]

Chemical formula

[0032] or a pharmaceutically acceptable salt thereof (wherein, X1, X2, and X3 are each independently C or N, provided that at least one of X1, X2, and X3 is N and at least one of X1, X2, and X3 is C; The dashed line “-” means a bond between X1 and X2 and between X2 and X3, which may be a single bond or a double bond, provided that at least one of the bonds between X1 and X2 and between X2 and X3 is a single bond; R 1 is absent, halogen, or C 1~6 alkyl, where said C 1~6 alkyl may optionally be mono-substituted or independently multi-substituted by hydroxyl, halogen, or deuterium; Each R 2 , R 3 and R 4 are independently absent, halogen, hydroxyl, cyano, C 1~6 alkyl, C 1~6 alkoxyl, -(CH2) n -Q, and optionally these are deuterium, hydroxyl, amino, cyano, halogen, C 1~6 alkyl, C 1~6 haloalkyl, (C≡N)-C 1~6 alkyl, C 1~6 alkoxyl, C 1~6 haloalkoxyl, C 3~8 cycloalkyl, C 3~8 cycloalkoxyl, a 3- to 8-membered aryl or a 3- to 8-membered heterocyclyl, and may be mono-substituted or independently multi-substituted therewith, where n is 0, 1, or 2, and Q is a 3- to 8-membered saturated or unsaturated carbocyclic ring or a 3- to 8-membered saturated or unsaturated heterocyclic ring; Ring A is a 5- to 12-membered aryl having 1 to 5 ring heteroatoms selected from oxygen, sulfur, and nitrogen, a 5- to 12-membered heteroaryl, or an 8- to 10-membered bicyclic ring having 0 to 5 ring heteroatoms selected from oxygen, sulfur, and nitrogen, where ring A is not phenyl) is provided.

[0033] In some embodiments, R2 is selected from methyl, ethyl, propyl, butyl, cyclopropyl, cyclobutyl, oxetanyl, cyclopentanyl, tetrahydrofuryl, cyclohexanyl, tetrahydropyranyl, cycloheptanyl, piperidinyl, phenyl, pyridinyl, pyridonyl, oxocanyl, tetrahydropyranyl, dihydropyranyl, spiro[3.3]heptanyl, spiro[2.5]octanyl, bicyclo[1.1.1]pentanyl, bicyclo[3.2.1]octanyl, 8-oxabicyclo[3.2.1]octan-3-yl, and optionally these are substituted with hydroxyl, cyano, halogen, C 1~6 alkyl, C 1~6 haloalkyl, C 1~6 alkoxyl, C 1~6 haloalkoxyl, C 3~8 cycloalkyl, C 3~8 cycloalkoxyl, a 3- to 8-membered aryl or a 3- to 8-membered heterocyclyl, which may be mono-substituted or independently multi-substituted, and optionally these are further substituted with halogen, deuterium, hydroxyl, amino, cyano, C 1~6 alkyl, C 1~6 haloalkyl, C 1~6 alkoxyl or C 1~6 haloalkoxyl, and may be mono-substituted or independently multi-substituted.

[0034] In some embodiments, R 2 is

[0035]

Chemical formula

[0036] selected from, and optionally these are mono-substituted or independently multi-substituted with hydroxyl, cyano, fluoro, chloro, bromo, methyl, ethyl, methoxyl, difluoromethyl, difluoromethoxyl or trifluoromethoxyl.

[0037] In some embodiments, R 2is cyclohexanyl or tetrahydropyranyl, and optionally these are monosubstituted or independently polysubstituted by halogen, C 1~6 alkyl or C 1~6 alkoxyl.

[0038] In some embodiments, R 1 is methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl or isobutyl, and optionally these are monosubstituted or independently polysubstituted by hydroxyl, halogen or deuterium.

[0039] In some embodiments, R 1 is methyl, ethyl, trifluoromethyl or trideuteriomethyl. In some embodiments, ring A is a 6-membered heteroaryl having one ring heteroatom of nitrogen, a 9-membered bicyclic ring having 2 to 3 ring heteroatoms selected from oxygen, sulfur and nitrogen, and optionally the 9-membered bicyclic ring is a phenyl or pyridinyl fused bicyclic ring, and optionally ring A is

[0040]

Chemical formula

[0041] selected from. In some embodiments, each R 3 and R 4 is independently absent, or selected from halogen, hydroxyl, cyano, C 1~6 alkyl, CN-C 1~6 alkyl, C 1~6 haloalkyl, C 1~6 alkoxyl, C 1~6 haloalkoxyl, a 3- to 8-membered saturated or unsaturated heterocyclyl, where the heterocyclyl is optionally further monosubstituted or independently polysubstituted by C 1~3 alkyl.

[0042] In some embodiments, ring A is

[0043]

Chemical formula

[0044] and each R 3 and R 4 is independently absent or selected from methyl, cyano, methoxyl, chloro, cyano-methyl, pyrazolyl, oxazolyl, where said pyrazolyl or oxazolyl may be further mono-substituted or independently poly-substituted by C 1~3 alkyl.

[0045] In some embodiments, the compounds provided by the present invention have the structure of formula Ia

[0046]

Chemical formula

[0047] and its pharmaceutically acceptable salts, wherein R1, R2, R3, R4, and ring A are as defined herein. In some embodiments, the compounds provided by the present invention have the structure of formula Ib

[0048]

Chemical formula

[0049] and its pharmaceutically acceptable salts, wherein R1, R2, R3, R4, and ring A are as defined herein. In some embodiments, the compounds provided by the present invention have the structure of formula Ic

[0050]

Chemical formula

[0051] and having the structure of its pharmaceutically acceptable salts, wherein R1, R2, R3, R4, and ring A are as defined herein. In some embodiments, the compounds provided by the present invention have the formula Id

[0052]

Chemical formula

[0053] and having the structure of its pharmaceutically acceptable salts, wherein R1, R2, R3, R4, and ring A are as defined herein. In some embodiments, the compounds provided by the present invention have the formula Ie

[0054]

Chemical formula

[0055] and having the structure of its pharmaceutically acceptable salts, wherein, one of X1 and X3 is N and the other is C, and the dashed line “-” represents the bond between X1 and N and between N and X3, which may be a single bond or a double bond, provided that at least one of the bonds between X1 and N and between N and X3 is a single bond; R 1 is C 1~3 alkyl, R 2 is cyclopentyl, cyclohexanyl, tetrahydropyranyl or 8-oxabicyclo[3.2.1]octan-3-yl, optionally substituted one or independently polysubstituted by halogen or C 1~3 alkoxyl, Y1, Y2 and Y3 are each independently C or N, provided that at least one of Y1, Y2 and Y3 is N; R 5 is halogen or C 1~3 alkyl, R 6 is C 1~3It is alkyl.

[0056] In some embodiments, R of formula Ie 2 is unsubstituted cyclopentyl, cyclohexanyl, tetrahydropyranyl or 8-oxabicyclo[3.2.1]octan-3-yl, optionally these are mono-substituted or independently polysubstituted by halogen or C 1~3 alkoxyl.

[0057] In some embodiments, Y3 of formula Ie is N, and at least one of Y1 and Y2 is N. In some embodiments, R of formula Ie 5 is methyl.

[0058] Representative compounds 1 to 149 of formula (I) are shown in Table 1 below.

[0059]

Table 1-1

[0060]

Table 1-2

[0061]

Table 1-3

[0062]

Table 1-4

[0063]

Table 1-5

[0064]

Table 1-6

[0065]

Table 1-7

[0066]

Table 1-8

[0067]

Table 1-9

[0068]

Table 1-10

[0069]

Table 1-11

[0070]

Table 1-12

[0071]

Table 1-13

[0072]

Table 1-14

[0073]

Table 1-15

[0074]

Table 1-16

[0075]

Table 1-17

[0076]

Table 1-18

[0077]

Table 1-19

[0078]

Table 1-20

[0079]

Table 1-21

[0080]

Table 1-22

[0081]

Table 1-23

[0082]

Table 1-24

[0083]

Table 1-25

[0084] It will be appreciated that some features of the present disclosure that have been described as separate embodiments for clarity may also be provided in combination as a single embodiment. Conversely, various features of the present disclosure that have been described as a single embodiment for brevity may also be provided separately or in any suitable sub-combination.

[0085] In various parts of the present disclosure, linking substituents are described. Where the structure clearly requires a linking group, the variable groups of the Markush formula listed as such groups are understood to be linking groups. For example, if the structure requires a linking group and the definition of the Markush group for the variable group lists "alkyl", then that "alkyl" is understood to represent a linking alkylene group.

[0086] As used herein, the term "substituted", when referring to a chemical group, means that the chemical group has one or more hydrogen atoms removed and replaced by a substituent. As used herein, the term "substituent" has its ordinary meaning known in the art and refers to a chemical moiety that is covalently bonded to the parent group or, where appropriate, fused thereto. As used herein, the term "optionally substituted" means that the chemical group may or may not have a substituent (i.e., is unsubstituted) or may have one or more substituents (i.e., is substituted). Substitution at a given atom should be understood to be limited by valence.

[0087] As used herein, the term "C i-j " indicates a range of carbon atom numbers, where i and j are integers, the range of carbon atom numbers includes the endpoints (i.e., i and j) and each integer point therebetween, and j is greater than i. For example, C 1~6 indicates a range of 1 to 6 carbon atoms, including 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, and 6 carbon atoms. In some embodiments, the term "C 1~12 " indicates 1 to 12, such as 1 to 10, 1 to 8, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 to 2 carbon atoms.

[0088] As used herein, the term "alkyl", whether used as part of another term or independently, refers to a saturated or unsaturated hydrocarbon chain, the latter of which can be further subdivided into hydrocarbon chains having at least one double or triple bond (alkenyl or alkynyl). In some embodiments, alkyl refers to a saturated hydrocarbon chain. The aforementioned hydrocarbon chains can be linear or branched. The term "C i-j alkyl" refers to an alkyl having i to j carbon atoms. Examples of saturated alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl; higher homologs such as 2-methyl-1-butyl, n-pentyl, 3-pentyl, n-hexyl, 1,2,2-trimethylpropyl, and the like. Examples of unsaturated alkyl groups include, but are not limited to, ethenyl, n-propenyl, isopropenyl, n-butenyl, sec-butenyl, ethynyl, propin-1-yl, propin-2-yl, and the like. "C 1~6 alkyl" examples include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, and tert-butyl. "C 1~3 alkyl" examples include, but are not limited to, methyl, ethyl, propyl, and isopropyl.

[0089] When representing an alkylene group to which "alkyl" is linked, examples of the alkylene group include, but are not limited to, methylene, 1,1-ethylene, 1,2-ethylene, 1,1-propylene, 1,2-propylene, 1,3-propylene, 2,2-propylene, tert-butanylene, and the like.

[0090] The term "amino" as used herein refers to a group of the formula "-NH2". As used herein, the term "carbamoyl" refers to an aminocarbonyl group (i.e., NH2-C(=O)-).

[0091] As used herein, the term "cyano" refers to a group of the formula "-C≡N". As used herein, the terms "halo" and "halogen" refer to fluoro, chloro, bromo or iodo groups.

[0092] As used herein, the term "hydroxyl" refers to a group of the formula "-OH". As used herein, the term "alkoxy", whether as part of another term or used independently, refers to a group of the formula -O-alkyl.

[0093] The term "C i-j alkoxy" means that the alkyl portion of the alkoxy group has i to j carbon atoms. Examples of alkoxy groups include, but are not limited to, methoxyl, ethoxyl, propoxyl (e.g., n-propoxyl and isopropoxyl), t-butoxyl, and the like. "C 1~12 alkoxyl" examples are methoxyl, ethoxyl and propoxyl.

[0094] As used herein, the term "hydroxyC 1~12 alkyl" refers to a group of the formula "-C 1~12 alkyl-OH", where the alkyl portion of this group has 1 to 12 carbon atoms and one or more hydroxyl groups may be linked to any carbon atom of the alkyl portion. In some embodiments, "C i-j alkyl-OH" has one hydroxyl group. Examples of "C 1~12 alkyl-OH" are hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl and 1-hydroxyisopropyl.

[0095] As used herein, the term "C i~j haloalkyl" refers to a (mono- or poly-substituted) C i~j alkyl group substituted with halogen. "C 1~12Examples of "haloalkyl" are fluoromethyl, difluoromethyl, trifluoromethyl, fluoroethyl, difluoroethyl, trifluoroethyl, chloroethyl, and bromoisopropyl. An example of "difluoroethyl" is 1,1-difluoroethyl. Examples of "trifluoroethyl" are 2,2,2-trifluoroethyl and 1,2,2-trifluoroethyl.

[0096] "C" i-j Examples of "haloalkoxyl" are fluoromethoxyl, difluoromethoxyl, or trifluoromethoxyl. Examples of "trifluoroethoxy" are 2,2,2-trifluoroethoxy and 1,2,2-trifluoroethoxy.

[0097] As used herein, the term "aryl" or "aromatic", whether used as part of another term or independently, refers to a ring system in which double bonds and single bonds alternate between atoms forming the ring. In the present disclosure, the term "aryl" or "aromatic" also means including pseudo-aromatic. The term "pseudo-aromatic" refers to a ring system that is not strictly aromatic but is stabilized by electron delocalization and exhibits behavior similar to an aromatic ring. An aryl or aromatic group may have a single ring or multiple rings. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, and the like.

[0098] As used herein, the term "heteroaryl" refers to an aryl containing at least one ring-forming heteroatom selected from O, S, N, P, etc. Heteroaryl includes, but is not limited to, furyl, thienyl, pyridinyl, triazinyl, pyridyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, indolizinyl, indolyl, isoindolyl, indolinyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,4-oxadiazol-5-one, 1,2,3-triazolyl, 1,3,4-thiadiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, quinazolinyl, isoquinazolinyl, 1,3,5-triazinyl, 1H thieno[2,3-c]pyrazolyl, thieno[2,3-b]furyl, 3H-indolyl, benzo[b]furanyl, benzo[b]thiophenyl, 1H-indazolyl, benzimidazolyl, tetrazolyl, uridinyl, and cytosinyl.

[0099] As used herein, the term "carbocyclic" refers to any ring, including monocyclic or polycyclic rings (e.g., having 2 or 3 fused, bridged, or spiro rings), all of whose ring atoms are carbon and which contain at least 3 ring-forming carbon atoms, whether used as part of another term or independently. As used herein, the term "spiro" ring refers to a ring system having two rings connected through a single common atom; the term "fused" ring refers to a ring system having two rings sharing two adjacent atoms; the term "bridged" ring refers to a ring system having two rings sharing more than 3 atoms.

[0100] In some embodiments, the carbocyclic ring may contain 3 to 12 ring-forming carbon atoms (i.e., 3- to 12-membered carbon atoms), 3 to 10 ring-forming carbon atoms, 3 to 9 ring-forming carbon atoms, or 3 to 8 ring-forming carbon atoms. The carbocyclic group may be saturated, partially unsaturated, or fully unsaturated. In some embodiments, the carbocyclic group may be a saturated cyclic alkyl group. In some embodiments, the carbocyclic group may be an unsaturated cyclic alkyl group containing at least one double bond within its ring system. In some embodiments, the unsaturated carbocyclic group may contain one or more aromatic rings. In some embodiments, one or more of the -CH2- groups forming one or more rings of the saturated or unsaturated carbocyclic may be replaced by a -C(O)- group.

[0101] In some embodiments, the carbocyclic group is a monocyclic alkyl group. In some embodiments, the carbocyclic group is a saturated monocyclic alkyl group. Examples of saturated monocyclic alkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopentenyl, cyclohexenyl, and the like.

[0102] A "3- to 8-membered saturated or unsaturated carbocyclic" is a saturated, partially unsaturated, or fully unsaturated monocyclic or polycyclic ring system having 3 to 8, 3 to 6, or 5 to 8 ring-forming carbon atoms, wherein one or more of the -CH2- groups forming one or more rings can optionally be replaced by a -C(O)- group.

[0103] Examples of "3- to 8-membered saturated or unsaturated carbocyclic" are C 3~6 cycloalkyl, cyclohexyl, cyclohexenyl, cyclopentyl, phenyl, naphthyl, and bicyclo[1.1.1]pentan-1-yl. Examples of "C 3~8 cycloalkyl" are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. The term "C 3~8 cycloalkoxyl" has the formula "C3~8 Refers to the group of "cycloalkyl - O -".

[0104] As used herein, the term "heterocyclyl" refers to a carbocyclic group in which one or more (e.g., 1, 2, or 3) ring atoms are replaced by heteroatoms including, but not limited to, O, S, N, P, etc. In some embodiments, the heterocyclyl is a saturated heterocyclyl. In some embodiments, the heterocyclyl is an unsaturated heterocyclyl having one or more double bonds within its ring system. In some embodiments, the heterocyclyl is a partially unsaturated heterocyclyl. In some embodiments, the heterocyclyl is a fully unsaturated heterocyclyl. In some embodiments, the unsaturated heterocyclyl group may contain one or more aromatic rings. In some embodiments, the - CH2 - groups forming one or more rings of the heterocyclyl can optionally be replaced by - C(O)-, - S -, - S(O)-, or - S(O)2 - groups. In some embodiments, when the heterocyclyl contains sulfur within its ring system, the sulfur atom forming the ring may optionally be oxidized to form an S - oxide. In some embodiments, the heterocyclyl is linked to another part of the compound through a carbon forming its ring. In some embodiments, the heterocyclyl is linked to another part of the compound through a nitrogen forming its ring.

[0105] In some embodiments, the 3 - to 8 - membered saturated or unsaturated monocyclic or polycyclic heterocyclyl has 1, 2, or 3 heteroatoms selected from N, O, or S.

[0106] "3 to 8-membered saturated or unsaturated heterocyclyl" each refers to a saturated, partially unsaturated or fully unsaturated monocyclic or polycyclic ring system (e.g., having 2 or 3 fused, bridged or spiro rings) with 3 to 8 ring-forming atoms, at least one of which is selected from nitrogen, sulfur or oxygen, and the ring system can be linked to other parts of the compound through carbon or nitrogen forming the ring, unless otherwise specified, where the -CH2- group forming one or more rings of the saturated or unsaturated heterocyclyl may be replaced by a -C(O)-, -S-, -S(O)-, or -S(O)2- group, and when the heterocyclyl contains sulfur in its ring system, the ring sulfur atom may optionally be oxidized to form an S-oxide.

[0107] Representative monocyclic heterocyclyl groups include, but are not limited to, oxetanyl, pyranyl, 1,1-dioxothietanyl pyrrolidyl, tetrahydrofuryl, tetrahydrothienyl, pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, triazolyl, oxazolyl, thiazolyl, piperidyl, piperidyl, piperazinyl, morpholinyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, pyridonyl, pyrimidonyl, pyrazinonyl, pyrimidonyl, pyridazinonyl, triazinonyl, etc.

[0108] Examples of spiroheterocyclyl include, but are not limited to, spiropyranyl, spirooxazinyl, and the like. Examples of fused heterocyclyl include, but are not limited to, phenyl-fused ring or pyridinyl-fused ring, such as quinolinyl, isoquinolinyl, quinoxalinyl, quinolidinyl, quinazolinyl, azaindolizinyl, pteridinyl, chromenyl, isochromenyl, indolyl, isoindolyl, indolizinyl, indazolyl, purinyl, benzofuranyl, isobenzofuranyl, benzimidazolyl, benzothienyl, benzothiazolyl, carbazolyl, phenazinyl, phenothiazinyl, phenanthridinyl, imidazo[1,2-a]pyridinyl, [1,2,4]triazolo[4,3-a]pyridinyl, [1,2,3]triazolo[4,3-a]pyridinyl group, and the like. Examples of bridged heterocyclyl include, but are not limited to, morphanyl, hexamethylenetetraminyl, 8-aza-bicyclo[3.2.1]octane, 1-aza-bicyclo[2.2.2]octane, 1,4-diazabicyclo[2.2.2]octane (DABCO), and the like.

[0109] The "compounds" of the present disclosure are intended to encompass all stereoisomers, geometric isomers, and tautomers of the depicted structures unless otherwise specified. The term "stereoisomer" refers to any of the various stereoisomeric arrangements (e.g., enantiomers, diastereomers, and racemates) of an asymmetric compound (e.g., one having one or more asymmetrically substituted carbon atoms or "asymmetric centers"). Compounds of the present disclosure containing an asymmetric center can be isolated in optically active (enantiomeric or diastereomeric) or optically inactive (racemic) forms. The term "enantiomer" includes a pair of stereoisomers that are mirror images that cannot be superimposed on one another. A 1:1 mixture of a pair of enantiomers is a "racemic mixture". The term "diastereomer" or "diastereoisomer" includes stereoisomers that have at least two asymmetric atoms but are not mirror images of one another. Certain compounds containing one or more asymmetric centers can give rise to enantiomers, diastereomers, or other stereoisomeric forms that can be defined as (R)- or (S)- according to the absolute configuration at each asymmetric center according to the Cahn-Ingold-Prelog R-S system. A resolved compound of unknown absolute configuration can be designated at the asymmetric center using the term "or". Methods for producing optically active forms from racemic mixtures are known in the art, such as resolution by HPLC or stereoselective synthesis.

[0110] The term "geometric isomer" or "cis-trans isomer" refers to compounds of the same formula, but whose functional groups are rotated in different directions in three-dimensional space. The term "tautomer" includes prototropic tautomers which are isomeric protonation states of a compound having the same formula and total charge. Examples of prototropic tautomers include, but are not limited to, keto-enol pairs, amide-imino acid pairs, lactam-lactim pairs, enamine-imine pairs, and cyclic forms in which a proton can occupy two or more positions of a heterocyclic system, such as 1H- and 3H-imidazole, 1H-, 2H- and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. Tautomers can be in equilibrium or can be sterically locked into one form by appropriate substitution. Compounds of the present disclosure identified as a particular tautomeric form by name or structure are intended to include other tautomeric forms unless otherwise specified.

[0111] The "compounds" of the present disclosure are also intended to encompass all isotopes of atoms in the compounds. Isotopes of an atom include atoms having the same atomic number but different mass numbers. For example, unless otherwise specified, hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, bromine or iodine in the "compounds" of the present disclosure are their isotopes, such as, but not limited to: 1 H, 2 H, 3 H, 11 C, 12 C, 13 C, 14 C, 14 N, 15 N, 16 O, 17 O, 18 O, 31 P, 32 P, 32 S, 33 S, 34 S, 36 S, 17 F, 19 F, 35 Cl, 37 Cl, 79 Br, 81 Br, 127 I and 131is meant to include I as well. In some embodiments, hydrogen includes protium, deuterium, and tritium. In some embodiments, the term "substituted by deuterium" or "substituted with deuterium" replaces other isotopes of hydrogen (e.g., protium) in a chemical group with deuterium. In some embodiments, carbon is 12 C and 13 C. In some embodiments, the "compounds" of the present disclosure only include isotopes of hydrogen in the compounds. In some embodiments, the "compounds" of the present disclosure only include isotopes of atoms in their natural abundance ratios.

[0112] Also, the "compounds" of the present disclosure can exist in solvated as well as non-solvated forms, such as hydrated forms, solid forms, and the present disclosure is to be understood as intending to encompass all such solvated and non-solvated forms.

[0113] Furthermore, it should be understood that the "compounds" of the present disclosure can exist in the form of pharmaceutically acceptable salts. As used herein, the term "pharmaceutically acceptable" refers to compounds, substances, compositions, and / or dosage forms that, within the scope of sound medical judgment, are suitable and proper for use in contact with human and animal tissues without excessive toxicity, irritation, allergic response, or other problems or complications and that exhibit a reasonable benefit / risk ratio. In some embodiments, pharmaceutically acceptable compounds, substances, compositions, and / or dosage forms are those that have been approved by a regulatory authority (e.g., the U.S. Food and Drug Administration, the China National Food and Drug Administration, or the European Medicines Agency) or are listed in widely recognized pharmacopoeias (e.g., the U.S. Pharmacopeia, the Chinese Pharmacopoeia, or the European Pharmacopoeia) for use in animals, more particularly in humans.

[0114] As used herein, "pharmaceutically acceptable salts" refers to derivatives of the compounds of the present disclosure that are modified by converting an acidic moiety (such as carboxyl, etc.) or a basic moiety (such as amine, alkali, etc.) in which the parent compound is present into its salt form. In many cases, the compounds of the present disclosure can form acid and / or base salts due to the presence of amino and / or carboxyl groups or groups similar thereto. Pharmaceutically acceptable salts are acid and / or base salts that generally retain the biological effectiveness and properties of the parent compound and are not undesirable in a biological or other sense. Suitable pharmaceutically acceptable salts of the compounds of the present disclosure include, for example, acid addition salts, which can be derived from, for example, inorganic acids (such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.) or organic acids (such as formic acid, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, trimellitic acid, citric acid, lactic acid, phenylacetic acid, benzoic acid, mandelic acid, methanesulfonic acid, napadisic acid, ethanesulfonic acid, toluenesulfonic acid, trifluoroacetic acid, salicylic acid, sulfosalicylic acid, etc.). In some embodiments, the pharmaceutically acceptable salt of the compound of the present disclosure is a formate. In some embodiments, the pharmaceutically acceptable salt of the compound of the present disclosure is a TFA salt.

[0115] Suitable pharmaceutically acceptable salts of the compounds of the present disclosure also include, for example, base addition salts, which can be derived from, for example, inorganic bases (e.g., sodium, potassium, ammonium salts and hydroxides, metals from columns I-XII of the periodic table, such as carbonates, bicarbonates of calcium, magnesium, iron, silver, zinc, copper, etc.) or organic bases (e.g., primary, secondary, and tertiary amines, substituted amines, such as naturally occurring substituted amines, cyclic amines, basic ion exchange resins, etc.). Certain organic amines include, but are not limited to, isopropylamine, benzathine, cholineate, diethanolamine, diethylamine, lysine, meglumine, piperazine, and tromethamine. As will be appreciated by those skilled in the art, it may also be possible to add acids or bases to form acid / base addition salts other than those shown in the examples. A list of additional suitable salts can be found, for example, in “Remington’s Pharmaceutical Sciences”, 20th ed., Mack Publishing Company, Easton, Pa., (1985) and “Handbook of Pharmaceutical Salts: Properties, Selection, and Use” by Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002). In some embodiments, suitable pharmaceutically acceptable salts of the compounds of the present disclosure are inorganic base salts.

[0116] The present disclosure also includes active intermediates, active metabolites, and prodrugs of the compounds of the present disclosure. As used herein, “active intermediate” refers to an intermediate compound that exhibits the same or essentially the same biological activity as the final compound synthesized in the synthetic process.

[0117] As used herein, "active metabolite" refers to a degradation or end product produced by the metabolism or biotransformation of a compound of the present disclosure, or a salt or prodrug thereof, in the body of an animal or human, which exhibits the same or essentially the same biological activity as the specific compound. Such metabolites can result from, for example, oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, enzymatic cleavage, etc. of the administered compound or salt or prodrug.

[0118] As used herein, "prodrug" refers to any compound or complex that releases an active parent drug when administered to an animal or human subject. Prodrugs are produced by modifying a functional group present in a compound such that the modification is cleavable from the parent compound either by routine manipulation or in vivo. Prodrugs include compounds in which a hydroxyl, amino, sulfhydryl, or carboxyl group is attached to a group that can cleave to form the free hydroxyl, amino, sulfhydryl, or carboxyl group, respectively, when administered to a mammalian subject. Examples of prodrugs include, but are not limited to, acetate, formate, and benzoate derivatives of alcohol and amine functional groups in the compounds of the present disclosure. The preparation and use of prodrugs are discussed in THiguchi and V. Stella, “Pro-drugs as Novel Delivery Systems”, Vol. 14 of the A.C.S. Symposium Series, and Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, both of which are hereby incorporated by reference in their entirety.

[0119] This specification discloses novel compounds or pharmaceutically acceptable salts thereof that can selectively inhibit DNA-PK. The compounds of the present disclosure or their pharmaceutically acceptable salts exhibit certain improved properties, such as higher BBB penetration (thus making them potentially useful for the treatment of cancers that have metastasized to the CNS, particularly brain metastases and leptomeningeal metastases), better efficacy, etc., when compared to other clinically available DNA-PK inhibitors. They may also possess a favorable toxicity profile and / or a favorable metabolic or pharmacokinetic profile compared to known DNA-PK inhibitors.

[0120] Accordingly, such compounds or their pharmaceutically acceptable salts are particularly useful for the treatment of cancer, particularly cancer having brain metastases. Synthesis methods The synthesis of the compounds of the present invention, including their salts, esters, hydrates, or solvates or stereoisomers, is illustrated in the synthesis schemes in the examples. The compounds provided by the present invention can be produced using any known organic synthesis techniques and can be synthesized according to any of a number of possible synthetic routes. Thus, these schemes are merely illustrative and are not meant to limit other possible methods that can be used to produce the compounds provided by the present invention. In addition, the steps of the schemes are for better illustration and can be changed as appropriate. Embodiments of the compounds in the examples were synthesized in China for the purposes of research and potentially for submission to regulatory authorities.

[0121] The reactions for producing the compounds of the present disclosure can be carried out in a suitable solvent that can be readily selected by those skilled in the art of organic synthesis. A suitable solvent can be substantially non-reactive with the starting materials (reactants), intermediates, or products at the temperature at which the reaction is carried out, for example, in the range from the freezing temperature of the solvent to the boiling temperature of the solvent. A given reaction can be carried out in one solvent or a mixture of more than one solvent. Depending on the particular reaction step, a solvent suitable for the particular reaction step can be selected by those skilled in the art.

[0122] The preparation of the compounds of the present disclosure can involve the protection and deprotection of various chemical groups. The need for protection and deprotection, as well as the selection of suitable protecting groups, can be readily determined by those skilled in the art. The chemistry of protecting groups can be found, for example, in T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3rd Ed., Wiley & Sons, Inc., New York (1999), which is hereby incorporated by reference in its entirety.

[0123] The reaction can be monitored according to any suitable method known in the art. For example, product formation can be monitored by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g., 1 H or 13 C), infrared spectroscopy, spectrophotometry (e.g., UV-visible), mass spectrometry, or by chromatography, such as high performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LCMS), or thin layer chromatography (TLC). Compounds can be purified by various methods including high performance liquid chromatography (HPLC) (“Preparative LC-MS Purification: Improved Compound Specific Method Optimization” Karl F. Blom, Brian Glass, Richard Sparks, Andrew P. Combs J. Combi. Chem. 2004, 6(6), 874~883, which is hereby incorporated by reference in its entirety) and normal phase silica chromatography by those skilled in the art.

[0124] The abbreviations used in this specification are defined as follows: "1×" or "×1" means once, "2×" or "×2" means twice, "3×" or "×3" means three times, "4×" or "×4" means four times, "5×" or "×5" means five times, "°C" means Celsius temperature, "eq" or "eq." means equivalent, "g" means gram, "mg" means milligram, "L" means liter, "mL" or "ml" means milliliter, "μL" means microliter, "N" means normal, "M" means molar concentration, "mmol" means millimole, "min" means minute, "h" or "hr" means hour, "r.t." or "rt" means room temperature, "atm" means atmospheric pressure, "psi" means pounds per square inch, "conc." means concentrate, "sat" or "sat’d" means saturated, "MS" or "Mass Spec" means mass spectrometry, "ESI" means electrospray ionization mass spectrometry, "LCMS" means liquid chromatography mass spectrometry, "HPLC" means high performance liquid chromatography, "RP" means reverse phase, "TLC" or "tlc" means thin layer chromatography, "SM" means starting material, "NMR" means nuclear magnetic resonance spectroscopy, " 1 "H" means proton, "δ" means delta, "s" means singlet, "d" means doublet, "t" means triplet, "q" means quartet, "m" means multiplet, "br" means broad, "Hz" means Hertz. "α", "β", "R", "S", "E", and "Z" are terms of stereochemistry familiar to those skilled in the art.

[0125] Pharmaceutical composition The present disclosure provides a pharmaceutical composition comprising at least one compound of the present disclosure. In some embodiments, the pharmaceutical composition comprises more than one compound of the present disclosure. In some embodiments, the pharmaceutical composition comprises one or more compounds of the present disclosure and a pharmaceutically acceptable carrier.

[0126] A pharmaceutically acceptable carrier is a conventional pharmaceutical carrier in the art that can be prepared by methods well known in the pharmaceutical field. In some embodiments, the compounds of the present disclosure can be mixed with a pharmaceutically acceptable carrier for the manufacture of a pharmaceutical composition.

[0127] As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable substance, composition or vehicle involved in transporting or delivering a compound provided by the present invention from one location, body fluid, tissue, organ (internal or external), or part of the body to another location, body fluid, tissue, organ, or part of the body, such as a liquid or solid filler, diluent, excipient, solvent or capsule material. A pharmaceutically acceptable carrier can be a vehicle, diluent, excipient, or other substance that can be used to contact animal tissue without undue toxicity or adverse effects. Representative pharmaceutically acceptable carriers include saccharides, starch, cellulose, malt, tragacanth, gelatin, Ringer's solution, alginic acid, isotonic saline, buffering agents, and the like. Pharmaceutically acceptable carriers that can be used in the present disclosure include those well known in the art, such as those disclosed in "Remington Pharmaceutical Sciences" Mack Pub. Co., New Jersey (1991), which is incorporated herein by reference.

[0128] Some examples of substances that can serve as pharmaceutically acceptable carriers are: (1) saccharides, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) tragacanth powder; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (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) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) alcohols, such as ethyl alcohol and propyl alcohol; (20) phosphate buffer solution; and (21) other non-toxic compatible substances used in pharmaceutical formulations, including acetone.

[0129] The pharmaceutical composition may contain pharmaceutically acceptable adjuvants required to approximate physiological conditions, such as pH regulators and buffering agents, toxicity regulators, etc., such as sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate, etc.

[0130] The form of the pharmaceutical composition is not limited, but depends on several criteria including the route of administration, the degree of the disease, or the amount of the drug administered. The pharmaceutical composition can be formulated for oral, intranasal, rectal, transdermal, intravenous, or intramuscular administration. For example, the dosage form for intranasal administration can be conveniently formulated as an aerosol, solution, nasal drops, gel, or dry powder; the dosage form for intranasal administration can be formulated as a fluid preparation. Depending on the desired route of administration, the pharmaceutical composition can be formulated in the form of tablets, capsules, pills, dragees, powders, granules, sachets, cachets, troches, suspensions, emulsions, solutions, syrups, aerosols (as solids or in a liquid medium), sprays, ointments, pastes, creams, lotions, gels, patches, inhalants, or suppositories.

[0131] The pharmaceutical composition can also be formulated to provide rapid, sustained or delayed release of the active ingredient after administration to the patient by using procedures known in the art. In some embodiments, the pharmaceutical composition is formulated in a sustained release form. As used herein, the term "sustained release form" refers to the release of an active pharmaceutical agent from a pharmaceutical composition of the active agent such that the active agent is available for biological absorption in the subject, primarily in the subject's gastrointestinal tract, over an extended period of time (extended release) or at a defined location (controlled release). In some embodiments, the extended period can be about 1 hour to 24 hours, 2 hours to 12 hours, 3 hours to 8 hours, 4 hours to 6 hours, 1 to 2 days or more. In certain embodiments, the extended period is at least about 4 hours, at least about 8 hours, at least about 12 hours, or at least about 24 hours. The pharmaceutical composition can be formulated in the form of tablets. For example, the release rate of the active agent can be controlled not only by the dissolution of the active agent in the gastrointestinal fluid and subsequent diffusion from the tablet or pill independent of pH, but also by the physical processes of tablet disintegration and erosion. In some embodiments, polymeric materials disclosed in "Medical Applications of Controlled Release," Langer and Wise (eds.), CRC Pres., Boca Raton, Florida (1974); "Controlled Drug Bioavailability," Drug Pproduct Design and Performance, Smolen and Ball (eds.), Wiley, New York (1984); Ranger and Peppas, 1983, J MacromolSci. Rev. Macromol Chem. 23:61; also Levy et al., 1985, Science 228:190; During et al., 1989, Ann. Neurol. 25:351; Howard et al., 1989, J. Neurosurg. 71:105 can be used for sustained release. The above references are incorporated herein by reference in their entirety.

[0132] In certain embodiments, the pharmaceutical composition comprises from about 0.0001 mg to about 100 mg of the compounds of the present disclosure (e.g., from about 0.0001 mg to about 10 mg, from about 0.001 mg to about 10 mg, from about 0.01 mg to about 10 mg, from about 0.1 mg to about 10 mg, from about 0.1 mg to about 5 mg, from about 0.1 mg to about 4 mg, from about 0.1 mg to about 3 mg, from about 0.1 mg to about 2 mg, from about 0.1 mg to about 1 mg, from about 0.1 mg to about 0.5 mg, from about 1 mg to about 10 mg, from about 1 mg to about 5 mg, from about 5 mg to about 10 mg, from about 5 mg to about 20 mg, from about 5 mg to about 30 mg, from about 5 mg to about 40 mg, from about 5 mg to about 50 mg, from about 10 mg to about 100 mg, from about 20 mg to about 100 mg, from about 30 mg to about 100 mg, from about 40 mg to about 100 mg, from about 50 mg to about 100 mg). Suitable dosages per day for a subject can be from about 0.1 mg to about 10 mg, preferably from about 0.1 mg to about 5 mg, from about 5 mg to about 10 mg, or from about 1 mg to about 5 mg.

[0133] In certain embodiments, the pharmaceutical composition can be formulated into unit dosage forms, and each dosage contains from about 0.0001 mg to about 10 mg, from about 0.001 mg to about 10 mg, from about 0.01 mg to about 10 mg, from about 0.1 mg to about 10 mg, from about 0.1 mg to about 5 mg, from about 0.1 mg to about 4 mg, from about 0.1 mg to about 3 mg, from about 0.1 mg to about 2 mg, from about 0.1 mg to about 1 mg, from about 0.1 mg to about 0.5 mg, from about 1 mg to about 10 mg, from about 5 mg to about 10 mg, from about 5 mg to about 20 mg, from about 5 mg to about 30 mg, from about 5 mg to about 40 mg, from about 5 mg to about 50 mg, from about 10 mg to about 100 mg, from about 20 mg to about 100 mg, from about 30 mg to about 100 mg, from about 40 mg to about 100 mg, from about 50 mg to about 100 mg of the compounds of the present disclosure. The term "unit dosage form" refers to physically discrete units suitable as unit dosages for human subjects and other mammals, and each unit contains a predetermined amount of the active substance calculated to produce the desired therapeutic effect together with a suitable pharmaceutical carrier.

[0134] In some embodiments, the pharmaceutical composition comprises one or more of the compounds of the present disclosure as a first active ingredient, and further comprises a second active ingredient. The second active ingredient can be any immunomodulatory agent or antitumor agent known in the art, including, without limitation, chemotherapy, immunotherapy, cell signaling inhibitors, alkylating agents, topoisomerase inhibitors, mitotic inhibitors, antihormonal agents, etc. Examples of such immunomodulatory agents or antitumor agents include platinum-based therapies (e.g., cisplatin (DDP), carboplatin (CBP), sulfato-1,2-diaminocyclohexane platinum (SHP), nedaplatin, oxaliplatin (OXA), lobaplatin), docetaxel, paclitaxel, doxorubicin, etoposide, mitoxantrone, CTLA-4 inhibitors, anti-CTLA-4 antibodies, PD-1 inhibitors, PD-L1 inhibitors, anti-PD-1 / PD-L1 antibodies, CD39 inhibitors, anti-CD39 antibodies, CD73 inhibitors, anti-CD73 antibodies, CCR2 inhibitors, anti-CCR2 antibodies, EGFR inhibitors, CDK4 / 6 inhibitors, MELK inhibitors, OX40 agonists, antiandrogen inhibitors, IgG4 isotype antibodies, tyrosine kinase inhibitors, DNA methyltransferase inhibitors, Hsp90 inhibitors, FGFR inhibitors, mTOR inhibitors, aromatase inhibitors, VEGF inhibitors, LHRH antagonists, PI3K inhibitors, AKT inhibitors, aurora kinase inhibitors, MEK inhibitors, HDAC inhibitors, BET inhibitors, PIK3CA inhibitors, proteasome inhibitors, other SERDs, farnesyl transferase inhibitors, VEGF-A antibodies, ErbB3 (Her3) antibodies, proteasome inhibitors, protein kinase Cβ inhibitors, anti-IGF-1R antibodies, anti-HER2 antibodies, SERMs, IGF inhibitors, anti-IgG antibodies, etc.Representative examples of antitumor agents for treating cancer or tumors include, but are not limited to, cisplatin, carboplatin, SHP, nedaplatin, oxaliplatin, lobaplatin, docetaxel, paclitaxel, doxorubicin, etoposide, mitoxantrone, vincristine, vinblastine, gemcitabine, cyclophosphamide, chlorambucil, carmustine, methotrexate, fluorouracil, actinomycin, epirubicin, anthracycline, bleomycin, mitomycin C, irinotecan, topotecan, teniposide, interleukin, interferon, tremelimumab, ipilimumab, pembrolizumab, nivolumab, avelumab, durvalumab, atezolizumab, IPH52, IPH53, CPI-006, prozartumab, MLN1202, cetuximab, lapatinib, erlotinib, gefitinib, neratinib, trastuzumab, trastuzumab emtansine, pertuzumab, MCLA-128, anastrozole, raloxifene, G1T38, tamoxifen, goserelin, enzalutamide, vorinostat, entinostat, sunitinib, pazopanib, bevacizumab, ranibizumab, pegaptanib, cediranib, dasatinib, GDC-0980, gedatolisib, alpelisib, BKM120, copanlisib, AZD8835, GDC-0941, taselisib, temsirolimus, everolimus, sapacitabine, AZD5363, MK2206, panitumumab, pembrolizumab, sorafenib, palbociclib, abemaciclib, ribociclib, crizotinib, dovitinib, luxitinib, azacitidine, CC-486, HSP90 ganetespib, Debio 1347, erdafitinib, vitusertib, alisertib, selumetinib, GS-5829, GSK525762, MLN9708, GDC-0810, AFP464, tipifarnib, seribantumab, bortezomib, enzastaurin, AVE1642, xentuzumab, daratumumab, AMG 479, etc.

[0135] Examples of such anti-tumor agents can also be found in Cancer Principles and Practice of Oncology by V. T. Devita and S. Hellman (editors), 6th edition (Feb. 15, 2001), Lippincott Williams & Wilkins Publishers. One of ordinary skill in the art will also be able to understand which combinations of agents are useful based on the individual properties of the drugs and the associated cancers.

[0136] According to this aspect of the disclosure, there is provided a suitable combination for use in the treatment of cancer, comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined above herein and any one of the immunomodulatory agents or anti-tumor agents listed above.

[0137] Thus, in a further aspect of the disclosure, there is provided a compound of formula (I), or a pharmaceutically acceptable salt thereof, in combination with an immunomodulatory agent or chemotherapy selected from those listed above.

[0138] As used herein, the term "combination" should be understood to refer to simultaneous, separate or sequential administration. In some embodiments, "combination" refers to simultaneous administration. In another aspect of the disclosure, "combination" refers to separate administration. In a further aspect of the disclosure, "combination" refers to sequential administration. When the administration is sequential or separate, the delay in administration of the second component should be such that the beneficial effects of the combination are not lost.

[0139] In a further aspect of the disclosure, there is provided a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, in combination with an immunomodulatory agent or anti-tumor agent selected from those listed above, together with a pharmaceutically acceptable diluent or carrier.

[0140] In a further aspect of the present disclosure, there is provided a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, selected from those listed above, in combination with an immunomodulatory agent or an antitumor agent for use in producing an immunomodulatory or anticancer effect, together with a pharmaceutically acceptable diluent or carrier.

[0141] In a further aspect of the present disclosure, there is provided a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, selected from those listed above, in combination with an immunomodulatory agent or an antitumor agent for use in treating DNA-PK related diseases such as NSCLC, RCC, prostate cancer or breast cancer, together with a pharmaceutically acceptable diluent or carrier.

[0142] In a further aspect of the present disclosure, there is provided a kit comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, in combination with an immunomodulatory agent or an antitumor agent selected from those listed above.

[0143] In a further aspect of the present disclosure, a) a compound of formula (I) or a pharmaceutically acceptable salt thereof in a first unit dosage form; b) an immunomodulatory agent or an antitumor agent in a second unit dosage form selected from those listed above; and c) a container containing said first and second dosage forms There is provided a kit comprising the same.

[0144] In addition to these uses in therapeutic agents, the compound of formula (I) or a pharmaceutically acceptable salt thereof is also useful as a pharmacological means in the development and standardization of in vitro and in vivo test systems for the evaluation of DNA-PK activity or expression in experimental animals such as cats, dogs, rabbits, monkeys, rats and mice as part of the search for novel therapeutic agents.

[0145] The other pharmaceutical compositions, processes, methods, uses, and features of the manufacture of medicaments described herein, alternative and preferred embodiments of the compounds of the present disclosure also apply. Method of treatment The present disclosure provides a method of treating a DNA-PK related disease, comprising administering to a subject an effective amount of one or more compounds of the present disclosure, pharmaceutically acceptable salts thereof, or pharmaceutical compositions.

[0146] The present disclosure also provides a method of treating a DNA-PK related disease. In certain embodiments, the method comprises administering to a subject an effective amount of one or more compounds of the present disclosure, pharmaceutically acceptable salts thereof, or pharmaceutical compositions.

[0147] As used herein, the term "DNA-PK related disease" refers to a disease in which the onset or progression or both are related to the expression or activity of DNA-PK. Examples include, but are not limited to, hyperproliferative diseases (e.g., cancer).

[0148] In some embodiments, the DNA-PK related disease is cancer, preferably DNA-PK overexpressing cancer. "DNA-PK overexpressing cancer" refers to those having a significantly higher level of DNA-PK protein in cancer or tumor cells as compared to non-cancerous cells of the same tissue type. Such overexpression can be caused by gene amplification or by increased transcription or translation. DNA-PK overexpression can be determined by assessing an increase in the level of DNA-PK protein present in cells in a diagnostic or prognostic assay (e.g., by immunohistochemical staining assay: IHC). Alternatively, and in addition, for example, fluorescence in situ hybridization (FISH; see WO98 / 45479, published October 1998), Southern blotting, or polymerase chain reaction (PCR) techniques, such as real-time quantitative PCR (RT-PCR) (Methods 132:73-80 (1990)), may be used to measure the level of nucleic acid encoding DNA-PK in cells. Apart from the above assays, various in vivo assays are available to those skilled in the art. For example, cells in a patient's body may be exposed to an antibody labeled, optionally, with a detectable label, such as a radioisotope, and the binding of the antibody to cells in the patient can be evaluated, for example, by external scanning for radioactivity or by analysis of a biopsy taken from the patient prior to exposure to the antibody.

[0149] In particular, cancer includes, but is not limited to, lung cancer (e.g., non-small cell lung cancer (NSCLC), small cell lung cancer, lung adenocarcinoma, large cell lung cancer, squamous cell lung cancer), renal cell carcinoma (RCC), prostate cancer, breast cancer, ovarian cancer, endometrial cancer, cervical cancer, bone cancer, uterine cancer, colorectal cancer, leukemia, glioblastoma, melanoma, chondrosarcoma, brain cancer, cholangiocarcinoma, osteosarcoma, lymphoma, adenoma, myeloma, hepatocellular carcinoma, adrenocortical carcinoma, pancreatic cancer, bladder cancer, liver cancer, gastric cancer, rectal cancer, esophageal cancer, testicular cancer, skin cancer, kidney cancer, mesothelioma, neuroblastoma, thyroid cancer, head and neck cancer, esophageal cancer, eye cancer, oropharyngeal cancer or oral cancer. In some embodiments, the cancer is NSCLC, RCC, prostate cancer or breast cancer. The cancer can be at any stage, unless otherwise specified, as described herein. In some embodiments, the cancer is early-stage cancer. In some embodiments, the cancer is locally advanced cancer. In some embodiments, the cancer is locally advanced and / or metastatic cancer. In some embodiments, the cancer is invasive cancer. In some embodiments, the cancer is cancer resistant to existing treatments.

[0150] As used herein, the terms “treatment,” “treating” and “treat” refer to reversing, reducing, delaying the onset of, or inhibiting the progression of a disease or disorder, or one or more symptoms thereof, as described herein. In some embodiments, treatment can be administered after one or more symptoms have appeared. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment can be administered to an individual predisposed to the onset of symptoms (e.g., taking into account a history of symptoms and / or genetic or other susceptibility factors). Treatment may also continue after symptoms have resolved, for example, to prevent or delay recurrence.

[0151] In some embodiments, the one or more compounds, pharmaceutically acceptable salts or pharmaceutical compositions provided by the present invention are administered by a parenteral route or a route other than parenteral. In some embodiments, the one or more compounds, pharmaceutically acceptable salts, hydrates, solvates or stereoisomers or pharmaceutical compositions are administered orally, enterally, buccally, nasally, intranasally, transmucosally, epidermally, transdermally, dermally, ophthalmically, pulmonary, rectally, sublingually, vaginally, topically, subcutaneously, intravenously, intramuscularly, intraarterially, intrathecally, intraarticularly, intraorbitally, intracardially, intradermally, intraperitoneally, intratracheally, subcuticularly, intraarticularly, subcapsularly, intrathecally, subarachnoidally, or intracisternally.

[0152] The compounds provided by the present invention can be administered in pure form, in combination with other active ingredients, or in the form of the pharmaceutical compositions disclosed herein. In some embodiments, the compounds provided by the present invention can be administered simultaneously or sequentially in combination with one or more anti-cancer or anti-inflammatory agents known in the art to the subject in need. The individual compounds of such combinations can be administered sequentially or simultaneously as separate or combined pharmaceutical compositions. Preferably, the individual compounds are administered simultaneously as a combined pharmaceutical composition. Suitable dosages of known therapeutic agents are readily understood by those skilled in the art.

[0153] In some embodiments, the administration is carried out once a day, twice a day, three times a day, or once every two days, once every three days, once every four days, once every five days, once every six days, once a week.

[0154] When a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof is provided herein, it will depend on various factors known in the art, such as body weight, age, past medical history, current medications, the health status of the subject and the potential for cross-reactivity, allergies, sensitivities and adverse side effects, as well as the route of administration and the extent of disease progression. The dosage may be proportionally decreased or increased by a person skilled in the art (e.g., a physician or veterinarian) as indicated by these or other circumstances or requirements.

[0155] In some embodiments, one or more compounds, pharmaceutically acceptable salts or pharmaceutical compositions provided by the present invention are administered orally. In the case of oral administration, any dosage amount that achieves the desired purpose is appropriate. In some embodiments, an appropriate daily dosage is about 0.001 - 100 mg, preferably 0.1 mg - 5 g, more preferably 5 mg - 1 g, even more preferably 10 mg - 500 mg, and the administration is once a day, twice a day, three times a day, daily, or 3 - 5 days a week. In some embodiments, the dosage amount of one or more compounds, pharmaceutically acceptable salts or pharmaceutical compositions provided by the present invention ranges from about 0.0001 mg, preferably 0.001 mg, 0.01 mg, 0.1 mg, 0.2 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.6 mg, 0.7 mg, 0.8 mg, 0.9 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg per day.

[0156] Use of the compound In certain embodiments, the present disclosure provides the use of a compound, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure in the manufacture of a medicament for treating DNA-PK related diseases. In certain embodiments, DNA-PK related diseases include cancer.

[0157] The compounds and pharmaceutical compositions in the present disclosure can be used for the prevention or treatment of the onset or progression (expression or activity) of any DNA-PK related disease in mammals, particularly humans.

[0158] In such a situation, the present disclosure also provides a method for screening a patient suitable for treatment with a compound or pharmaceutical composition of the present disclosure, alone or in combination with other components (e.g., a second active ingredient, e.g., an anti-inflammatory or anti-cancer agent). The method includes arranging tissue samples from the patient and detecting the accumulation of DNA-PK in the patient.

Examples

[0159] The following further illustrates the general method of the present disclosure. The compounds of the present disclosure can be produced by methods known in the art. The following illustrates a detailed production method of the preferred compounds of the present disclosure. However, the following does not limit the production method of the compounds of the present disclosure in any sense.

[0160] Synthesis Examples The synthesis of the compounds provided by the present invention, including their pharmaceutically acceptable salts, is illustrated in the synthesis schemes in the examples. The compounds provided by the present invention can be prepared using any known organic synthesis techniques and can be synthesized according to any of a number of possible synthetic routes. Therefore, these schemes are only illustrative and are not meant to limit other possible methods that can be used to prepare the compounds provided by the present invention. In addition, the steps in the schemes are for better explanation and can be changed as appropriate. The embodiments of the compounds in the examples were synthesized for the purposes of research and potentially for submission to regulatory authorities.

[0161] The reactions for preparing the compounds of the present disclosure can be carried out in a suitable solvent that can be easily selected by those skilled in the art of organic synthesis. A suitable solvent can be substantially non-reactive with the starting materials (reactants), intermediates, or products at the temperature at which the reaction is carried out, for example, a temperature within the range from the freezing temperature of the solvent to the boiling temperature of the solvent. A given reaction can be carried out in one solvent or a mixture of more than one solvent. Depending on the individual reaction steps, a suitable solvent for each individual reaction step can be selected by a skilled person.

[0162] The preparation of the compounds of the present disclosure can involve the protection and deprotection of various chemical groups. The need for protection and deprotection, as well as the selection of appropriate protecting groups, can be readily determined by those skilled in the art. The chemistry of protecting groups can be found, for example, in T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3 rd Ed., Wiley & Sons, Inc., New York (1999), which is hereby incorporated by reference in its entirety.

[0163] The reaction can be monitored according to any suitable method known in the art. For example, the formation of the product can be monitored by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g., 1 H or 13 C), infrared spectroscopy, spectrophotometry (e.g., UV-visible), mass spectrometry, or by chromatography, such as high performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LCMS), or thin layer chromatography (TLC). The compounds can be purified by those skilled in the art by various methods, such as high performance liquid chromatography (HPLC) (“Preparative LC-MS Purification: Improved Compound Specific Method Optimization” Karl F. Blom, Brian Glass, Richard Sparks, Andrew P. Combs J. Combi. Chem. 2004, 6(6), 874~883, which is hereby incorporated by reference in its entirety), and normal phase silica chromatography.

[0164] The structures of the compounds in the examples are characterized by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS). The chemical shifts (δ) of NMR are expressed in units of -6 (ppm). 11H-NMR spectra were recorded using ICON-NMR (under TopSpin program control) with tetramethylsilane as the internal standard on a Bruker AVANCE NMR (300 MHz or 400 MHz) spectrometer in dimethyl sulfoxide-d6 (DMSO-d6) or CDCl3 or CD3OD or D2O or acetone-d6 or CD3CN (Innochem or Sigma-Aldrich or Cambridge Isotope Lab., Inc.).

[0165] MS measurements were performed using a Shimadzu 2020 mass spectrometer equipped with an electrospray source in positive and negative ion modes. High performance liquid chromatography (HPLC) measurements were carried out on a Shimadzu LC-20AD system or a Shimadzu LC-20ADXR system or a Shimadzu LC-30AD system using a Shim-pack XR-ODS C18 column (3.0×50 mm, 2.2 μm), or an Ascentis Express C18 column (2.1×50 mm, 2.7 μm), or an Agilent Poroshell HPH-C18 column (3.0×50 mm, 2.7 μm).

[0166] Thin layer chromatography was performed using silica gel plates from Sinopharm Chemical Reagent Beijing Co., Ltd. and Xinnuo Chemical. The silica gel plates used for thin layer chromatography (TLC) were 175 - 225 μm. The silica gel plates used to separate and purify the product by TLC were 1.0 mm.

[0167] The purified chromatography column uses silica gel (100 - 200, 200 - 300 or 300 - 400 mesh, produced by Rushanshi Shangbang Xincailiao Co., Ltd. or Rushan Taiyang Desiccant Co., Ltd., etc.) as the carrier, or a flash column (reverse-phase C18 column 20 - 45μm, produced by Agela Technologies) in the Agela Technologies flash system. The size of the column is adjusted according to the amount of the compound.

[0168] The known starting materials of the present disclosure can be synthesized using or according to methods known in the art, or can be purchased from Alfa Aesar, TCI, Sigma-Aldrich, Bepharm, Bide pharmatech, PharmaBlock, Enamine, Innochem and JW&Y PharmLab, etc.

[0169] Unless otherwise specified, all reactions are carried out under an argon or nitrogen atmosphere. An argon or nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon with a volume of about 1L. Hydrogenation is usually carried out under normal pressure. Unless otherwise specified, the reaction temperature in the examples is ambient temperature, which is 10°C - 30°C. The progress of the reaction is monitored by TLC or / and LC-MS. The eluent systems used in the reaction include dichloromethane-methanol system and petroleum ether-ethyl acetate system. The volume ratio of the solvents is adjusted according to the different polarities of the compounds.

[0170] The eluent system for column chromatography and the eluent system for TLC used to purify the compound include dichloromethane-methanol system and petroleum ether-ethyl acetate system. The volume ratio of the solvents is adjusted according to the different polarities of the compounds. A small amount of alkaline or acidic substances (0.1% - 1%), such as formic acid, or acetic acid, or TFA, or ammonia can be added for adjustment.

[0171] Abbreviations of chemical substances used in the synthesis of the compounds provided by the present invention are listed below:

[0172] [Table 2]

[0173] Example 1 Preparation of 1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-3-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 1)

[0174] [Chemical formula]

[0175] Step 1. 5-chloro-3-iodo-1H-pyrazolo[4,3-d]pyrimidine A mixture of 5-chloro-1H-pyrazolo[4,3-d]pyrimidine (3.00 g, 19.410 mmol, 1.00 equivalent) and NIS (7.86 g, 34.936 mmol, 1.80 equivalents) in DMF (60.00 mL) was stirred overnight under an air atmosphere at 0 °C. The resulting mixture was extracted with EtOAc (3 × 150 mL). The combined organic layers were washed with brine (3 × 200 mL). The combined organic layers were washed with Na2S2O3 (3 × 200 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with PE / EtOAc (20:1) to give 5-chloro-3-iodo-1H-pyrazolo[4,3-d]pyrimidine (2.4 g, 44.09%). LCMS: m / z (ESI), [M+H] + = 281.0.

[0176] Step 2. 5-chloro-3-iodo-1-methyl-1H-pyrazolo[4,3-d]pyrimidine A mixture of Cs2CO3 (3.49 g, 10.697 mmol, 3 equiv), CH3I (2.53 g, 17.828 mmol, 5.00 equiv) and 5-chloro-3-iodo-1H-pyrazolo[4,3-d]pyrimidine (1.00 g, 3.566 mmol, 1.00 equiv) in DMF (20.00 mL) was stirred for 1 h at 0 °C under a nitrogen atmosphere. The resulting mixture was diluted with water (100 mL) and extracted with EtOAc (3 × 80 mL). The combined organic layers were washed with brine (3 × 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was recrystallized from EtOAc / PE (1:5 300 mL) to give 5-chloro-3-iodo-1-methyl-1H-pyrazolo[4,3-d]pyrimidine (850 mg, 80.95%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 295.0。

[0177] Step 3. 5-Chloro-3-(3,6-dihydro-2H-pyran-4-yl)-1-methyl-1H-pyrazolo[4,3-d]pyrimidine K2CO3 (1210.85 mg, 8.761 mmol, 3.00 equiv), Pd(dppf)Cl2CH2Cl2 (476.98 mg, 0.584 mmol, 0.2 equiv), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (797.57 mg, 3.797 mmol, 1.3 equiv) and 5-chloro-3-iodo-1-methyl-1H-pyrazolo[4,3-d]pyrimidine (860.00 mg, 2.920 mmol, 1.00 equiv) in dioxane (15.00 mL) and H2O (3.00 mL) were stirred at 80 °C for 16 h under a nitrogen atmosphere. The resulting mixture was diluted with water (100 mL). The resulting mixture was extracted with CH2Cl2 / MeOH = 12:1 (3 × 50 mL). The combined organic layers were washed with brine (2 × 100 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 12:1) to give 5-chloro-3-(3,6-dihydro-2H-pyran-4-yl)-1-methyl-1H-pyrazolo[4,3-d]pyrimidine (380 mg, 51.90%) as a gray solid. LCMS: m / z (ESI), [M+H] + = 251.2。

[0178] Step 4. 3-(3,6-Dihydro-2H-pyran-4-yl)-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine A mixture of Cs2CO3 (2599.38 mg, 7.978 mmol, 2.50 equiv), Xantphos (553.94 mg, 0.957 mmol, 0.30 equiv), Pd(OAc)2 (143.29 mg, 0.638 mmol, 0.20 equiv), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (567.40 mg, 3.829 mmol, 1.20 equiv) and 5-chloro-3-(3,6-dihydro-2H-pyran-4-yl)-1-methyl-1H-pyrazolo[4,3-d]pyrimidine (800.00 mg, 3.191 mmol, 1.00 equiv) in dioxane (20.00 mL) was stirred overnight at 100 °C under a nitrogen atmosphere. The resulting mixture was diluted with water (200 mL). The resulting mixture was extracted with CH2Cl2 / MeOH = (12:1) (3 × 200 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was recrystallized from EtOAc / PE (1:6 300 mL) to give 3-(3,6-dihydro-2H-pyran-4-yl)-1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1H-pyrazolo[4,3-d]pyrimidin-5-amine (600 mg, 51.88%) as a brown solid. LCMS: m / z (ESI), [M+H] + = 363.3。

[0179] Step 5. 1-Methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-3-(oxan-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 1) A mixture of Pd / C (47.92 mg, 0.450 mmol, 1.36 equiv) and 3-(3,6-dihydro-2H-pyran-4-yl)-1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1H-pyrazolo[4,3-d]pyrimidin-5-amine (120 mg, 0.331 mmol, 1.00 equiv) in MeOH (200 mL) and THF (100 mL) was stirred for 2 h under a hydrogen atmosphere at room temperature. The resulting mixture was filtered and the filter cake was washed with MeOH (3×100 mL). The filtrate was concentrated under reduced pressure. The crude product (120 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30×150 mm, 5 um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 25B~51B, 7 min) to give 1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-3-(oxan-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (55 mg, 45.12%) as a white solid. LCMS: m / z (ESI), [M+H] + =365.2。 1 H NMR (300 MHz, DMSO-d6) δ 1.95 (4H, t), 2.42 (3H, d), 3.14 - 3.30 (1H, m), 3.47 (2H, d), 3.93 (2H, d), 4.04 (3H, s), 7.71 (1H, t), 8.37 (1H, s), 8.84 (1H, s), 9.15 (1H, s), 9.34 (1H, s) Example 2 Preparation of 1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-3-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 2) and 3-(3,6-dihydro-2H-pyran-4-yl)-1-methyl-N-[7-methylimidazo[1,2-a]pyridin-6-yl]pyrazolo[4,3-d]pyrimidin-5-amine (Example 6)

[0180] [Chemistry]

[0181] Step 1. 3-(3,6-Dihydro-2H-pyran-4-yl)-1-methyl-N-[7-methylimidazo[1,2-a]pyridin-6-yl]pyrazolo[4,3-d]pyrimidin-5-amine (Example 6) A mixture of Cs2CO3 (682.34 mg, 2.094 mmol, 2.50 eq), Xantphos (96.94 mg, 0.168 mmol, 0.20 eq), Pd(OAc)2 (37.61 mg, 0.168 mmol, 0.20 eq), 7-methylimidazo[1,2-a]pyridin-6-amine (147.95 mg, 1.005 mmol, 1.20 eq) and 5-chloro-3-(3,6-dihydro-2H-pyran-4-yl)-1-methylpyrazolo[4,3-d]pyrimidine (210.00 mg, 0.838 mmol, 1.00 eq) in dioxane (6.00 mL) was stirred overnight at 100 °C under a nitrogen atmosphere. The desired product could be detected by LCMS. The resulting mixture was filtered and the filter cake was washed with DCM (3 × 50 mL). The filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 12:1) to give the crude product. The crude product (170 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column 19*250 mm, 5um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 26B~36B, 7 min) to give 3-(3,6-dihydro-2H-pyran-4-yl)-1-methyl-N-[7-methylimidazo[1,2-a]pyridin-6-yl]pyrazolo[4,3-d]pyrimidin-5-amine (100 mg, 57.65%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 362.2. 11H NMR (300 MHz, DMSO-d6) δ 2.30 (3H, d), 2.58 (2H, s), 3.83 (2H, t), 4.06 (3H, s), 4.25 (2H, d), 7.07-7.12 (1H, m), 7.43 (1H, d), 7.49 (1H, d), 7.84 (1H, t), 8.81 (2H, d), 9.14 (1H, s) Step 2. 1-Methyl-N-[7-methylimidazo[1,2-a]pyridin-6-yl]-3-(oxan-4-yl)pyrazolo[4,3-d]pyrimidin-5-amine (Example 2) A mixture of Pd / C (70.67 mg, 0.664 mmol, 3.00 equiv) and 3-(3,6-dihydro-2H-pyran-4-yl)-1-methyl-N-[7-methylimidazo[1,2-a]pyridin-6-yl]pyrazolo[4,3-d]pyrimidin-5-amine (80.00 mg, 0.221 mmol, 1.00 equiv) in MeOH (20.00 mL) was stirred for 3 h under a hydrogen atmosphere at 40 °C. The desired product could be detected by LCMS. The resulting mixture was filtered and the filter cake was washed with MeOH (3 × 10 mL). The filtrate was concentrated under reduced pressure. The crude product (50 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column 19*250 mm, 5 um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 22B~33B, 7 min RT1: 6.63) to obtain 1-methyl-N-[7-methylimidazo[1,2-a]pyridin-6-yl]-3-(oxan-4-yl)pyrazolo[4,3-d]pyrimidin-5-amine (20 mg, 24.61%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 364.2 11H NMR (300 MHz, DMSO-d6) δ 1.96 (4H, d), 2.28 (3H, d), 3.21 (1H, t), 3.47 (2H, d), 3.93 (2H, d), 4.03 (3H, s), 7.42 (1H, q), 7.49 (1H, d), 7.81 (1H, t), 8.69 (1H, s), 8.84 (1H, s), 9.09 (1H, s) Example 3 Preparation of 7-methyl-N-(5-methyl-7-(tetrahydro-2H-pyran-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)-[1,2,4]triazolo[1,5-a]pyridin-6-amine (Example 3)

[0182]

Chemical formula

[0183] Step 1. 2-chloro-7-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-5H-pyrrolo[3,2-d]pyrimidine A mixture of 2-chloro-7-iodo-5-methylpyrrolo[3,2-d]pyrimidine (300.00 mg, 1.022 mmol, 1.00 equivalent), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (279.16 mg, 1.329 mmol, 1.30 equivalent), Pd(dppf)Cl2 (149.59 mg, 0.204 mmol, 0.20 equivalent) and K2CO3 (423.81 mg, 3.067 mmol, 3.00 equivalent) in dioxane (6.00 mL) and H2O (1.20 mL) was stirred for 3 hours at 80 °C under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (PE / EtOAc 1:3) to give 2-chloro-7-(3,6-dihydro-2H-pyran-4-yl)-5-methylpyrrolo[3,2-d]pyrimidine (196 mg, 76.79%) as a brown solid. LCMS: m / z (ESI), [M+H] + = 250.2

[0184] Step 2. 7-(3,6-Dihydro-2H-pyran-4-yl)-5-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrrolo[3,2-d]pyrimidin-2-amine A mixture of 2-chloro-7-(3,6-dihydro-2H-pyran-4-yl)-5-methylpyrrolo[3,2-d]pyrimidine (196.00 mg, 0.785 mmol, 1.00 equiv), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (139.56 mg, 0.942 mmol, 1.20 equiv), Pd(AcO)2 (35.25 mg, 0.157 mmol, 0.20 equiv), Xantphos (136.25 mg, 0.235 mmol, 0.30 equiv) and Cs2CO3 (639.37 mg, 1.962 mmol, 2.50 equiv) in dioxane (3.00 mL) was stirred at 100 °C for 3 h under a nitrogen atmosphere. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 10:1) to give 7-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrrolo[3,2-d]pyrimidin-2-amine (170 mg, 59.93%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 362.3.

[0185] Step 3. 7-Methyl-N-(5-methyl-7-(tetrahydro-2H-pyran-4-yl)-5H-pyrrolo[3,2-d]pyrimidin-2-yl)-[1,2,4]triazolo[1,5-a]pyridin-6-amine (Example 3) A mixture of 7-(3,6-dihydro-2H-pyran-4-yl)-5-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrrolo[3,2-d]pyrimidin-2-amine (170.00 mg, 0.470 mmol, 1.00 equiv) and Pd / C (250.29 mg, 2.352 mmol, 5.00 equiv) in MeOH (20.00 mL) and THF (50.00 mL) was stirred overnight under a hydrogen atmosphere at room temperature. The resulting mixture was filtered and the filter cake was washed with MeOH (5 × 30 mL). The filtrate was concentrated under reduced pressure to give the crude product. The crude product (100 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column 19*250 mm, 5um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 25 mL / min; gradient: 30B~45B, 7 min; RT1: 6.02) to give 5-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-7-(oxan-4-yl)pyrrolo[3,2-d]pyrimidin-2-amine (29 mg, 16.96%) as a white solid. LCMS: m / z (ESI), [M+H] + = 364.3。 1 H NMR (300 MHz, DMSO-d6) δ 1.76 (2H, d), 1.88 - 2.01 (2H, m), 2.46 (3H, d), 3.02 (1H, t), 3.47 (2H, td), 3.80 (3H, s), 3.88 - 4.05 (2H, m), 7.51 (1H, s), 7.61 - 7.75 (1H, m), 8.33 (2H, d), 8.74 (1H, s), 9.49 (1H, s) Example 4 Preparation of 3-isopropyl-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 4)

[0186]

Chemical Structure

[0187] Step 1. 5-Chloro-1-methyl-3-(prop-1-en-2-yl)-1H-pyrazolo[4,3-d]pyrimidine A mixture of Pd(dppf)Cl2 (124.24 mg, 0.170 mmol, 0.20 equiv), K2CO3 (293.33 mg, 2.122 mmol, 2.50 equiv), 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (213.99 mg, 1.273 mmol, 1.50 equiv) and 5-chloro-3-iodo-1-methylpyrazolo[4,3-d]pyrimidine (250.00 mg, 0.849 mmol, 1.00 equiv) in dioxane (5.00 mL) and H2O (1.00 mL) was stirred for 2 h at 80 °C under a nitrogen atmosphere. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (PE / EtOAc 1:3) to give 5-chloro-1-methyl-3-(prop-1-en-2-yl)pyrazolo[4,3-d]pyrimidine (130 mg, 73.39%) as a pink solid. LCMS: m / z (ESI), [M+H] + = 209.6. 1 H NMR (300 MHz, DMSO-d6) δ 2.22 (3H, t), 4.18 (3H, s), 5.50 (1H, p), 6.43 (1H, d), 9.45 (1H, s) Step 2. 1-Methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-3-(prop-1-en-2-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine A mixture of Cs2CO3 (468.47 mg, 1.438 mmol, 2.50 equiv), Xantphos (66.56 mg, 0.115 mmol, 0.20 equiv), Pd(AcO)2 (25.82 mg, 0.115 mmol, 0.20 equiv), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (127.82 mg, 0.863 mmol, 1.50 equiv) and 5-chloro-1-methyl-3-(prop-1-en-2-yl)pyrazolo[4,3-d]pyrimidine (120.00 mg, 0.575 mmol, 1.00 equiv) in dioxane (4.00 mL) was stirred for 3 h at 100 °C under a nitrogen atmosphere. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 15:1) to give 1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-3-(prop-1-en-2-yl)pyrazolo[4,3-d]pyrimidin-5-amine (90 mg, 48.85%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 321.3。 1 H NMR (300 MHz, DMSO-d6) δ 2.18 (3H, s), 2.45 (3H, d), 4.09 (3H, d), 5.33 (1H, d), 6.35 (1H, d), 7.70 - 7.78 (1H, m), 8.38 (1H, s), 8.96 (1H, s), 9.22 (1H, s), 9.37 (1H, s) Step 3. 3-Isopropyl-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 4) A mixture of Pd / C (89.69 mg, 0.843 mmol, 3.00 equiv) and 1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-3-(prop-1-en-2-yl)pyrazolo[4,3-d]pyrimidin-5-amine (90.00 mg, 0.281 mmol, 1.00 equiv) in MeOH (10.00 mL) was stirred overnight under a hydrogen atmosphere at room temperature. The desired product could be detected by LCMS. The resulting mixture was filtered and the filter cake was washed with MeOH (4 × 50 mL). The filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2:MeOH 12:1) to give the crude product. The crude product (80 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column 19*250 mm, 5 um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: MeOH; flow rate: 25 mL / min; gradient 58B~70B, 7 min; RT1 5.57) to give 3-isopropyl-1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[4,3-d]pyrimidin-5-amine (50 mg, 54.66%) as a white solid. LCMS: m / z (ESI), [M+H] + = 323.2. 1 H NMR (300 MHz, DMSO-d6) δ 1.39 (6H, d), 2.45 (3H, d), 3.18 - 3.34 (1H, m), 4.04 (3H, s), 7.69 - 7.75 (1H, m), 8.37 (1H, s), 8.79 (1H, s), 9.16 (1H, s), 9.44 (1H, s) Example 5 Preparation of 3-cyclohexyl-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 5)

[0188]

Chemical Structure

[0189] Step 1. 5-Chloro-3-(cyclohex-1-en-1-yl)-1-methyl-1H-pyrazolo[4,3-d]pyrimidine K2CO3 (293.33 mg, 2.122 mmol, 2.50 equiv), 2-(cyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (265.01 mg, 1.273 mmol, 1.50 equiv) and 5-chloro-3-iodo-1-methylpyrazolo[4,3-d]pyrimidine (250.00 mg, 0.849 mmol, 1.00 equiv) in dioxane (5.00 mL) and H2O (1.00 mL) were stirred under a nitrogen atmosphere at 80 °C for 2 h. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (PE / EtOAc 1:2) to give 5-chloro-3-(cyclohex-1-en-1-yl)-1-methylpyrazolo[4,3-d]pyrimidine (150 mg, 71.04%) as a pink solid. LCMS: m / z (ESI), [M+H] + = 249.3 1 H NMR (300 MHz, DMSO-d6) δ 1.61 - 1.80 (4H, m), 2.29 (2H, s), 2.54 (2H, s), 4.14 (3H, s), 7.17 - 7.27 (1H, m), 9.40 (1H, s) Step 2. 3-(Cyclohex-1-en-1-yl)-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine A mixture of Cs2CO3 (458.51 mg, 1.407 mmol, 2.50 equiv), Xantphos (65.14 mg, 0.113 mmol, 0.20 equiv), Pd(AcO)2 (25.28 mg, 0.113 mmol, 0.20 equiv), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (125.11 mg, 0.844 mmol, 1.50 equiv) and 5-chloro-3-(cyclohex-1-en-1-yl)-1-methylpyrazolo[4,3-d]pyrimidine (140.00 mg, 0.563 mmol, 1.00 equiv) in dioxane (4.00 mL) was stirred for 3 h at 100 °C under a nitrogen atmosphere. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 15:1) to give 3-(cyclohex-1-en-1-yl)-1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[4,3-d]pyrimidin-5-amine (140 mg, 69.00%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 361.3. 1 H NMR (300 MHz, DMSO-d6) δ 1.61 - 1.83 (4H, m), 2.17 - 2.33 (4H, m), 2.46 (3H, d), 4.06 (3H, s), 7.21 (1H, s), 7.73 (1H, s), 8.38 (1H, s), 8.88 (1H, s), 9.19 (1H, s), 9.46 (1H, s) Step 3. 3-Cyclohexyl-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 5) A solution of 3-(cyclohex-1-en-1-yl)-1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[4,3-d]pyrimidin-5-amine (160.00 mg, 0.444 mmol, 1.00 equiv) and Pd / C (236.21 mg, 2.220 mmol, 5.00 equiv) in THF (40.00 mL) and MeOH (80.00 mL) was stirred for 3 days under a hydrogen atmosphere at room temperature. The precipitated solid was collected by filtration and washed with MeOH (5 × 30 mL). The resulting mixture was concentrated under reduced pressure. The crude product (150 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column 30 × 150 mm, 5 um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 40B~50B, 7 min; RT1: 6.55) to give 3-cyclohexyl-1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[4,3-d]pyrimidin-5-amine (17.41 mg, 10.82%) as a white solid. LCMS: m / z (ESI), [M+H] + = 363.2. 1 H NMR (300 MHz, DMSO-d6) δ 1.33 (3H, d), 1.76 (5H, d), 2.00 (2H, d), 2.46 (3H, d), 2.88 - 3.05 (1H, m), 4.04 (3H, s), 7.72 (1H, t), 8.38 (1H, s), 8.78 (1H, s), 9.15 (1H, s), 9.51 (1H, s) Example 8 Preparation of 3-((1r,4r)-4-methoxycyclohexyl)-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 7) and 3-((1s,4s)-4-methoxycyclohexyl)-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 8)

[0190] [Chemistry]

[0191] Step 1. 5-Chloro-3-(4-methoxycyclohex-1-en-1-yl)-1-methyl-1H-pyrazolo[4,3-d]pyrimidine A mixture of K2CO3 (234.66 mg, 1.698 mmol, 2.50 equiv), Pd(dppf)Cl2CH2Cl2 (110.93 mg, 0.136 mmol, 0.20 equiv), 2-(4-methoxycyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (194.07 mg, 0.815 mmol, 1.20 equiv) and 5-chloro-3-iodo-1-methylpyrazolo[4,3-d]pyrimidine (200.00 mg, 0.679 mmol, 1.00 equiv) in dioxane (5.00 mL) and H2O (1.00 mL) was stirred for 2 h at 80 °C under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (PE / EtOAc 1:2) to give 5-chloro-3-(4-methoxycyclohex-1-en-1-yl)-1-methylpyrazolo[4,3-d]pyrimidine (175 mg, 92.44%) as a white solid. LCMS: m / z (ESI), [M+H] + = 279.3. 1 H NMR (300 MHz, CDCl3) δ 1.89 (2H, d), 2.11 (1H, m), 2.33 (1H, t), 2.66 (1H, m), 2.88 (1H, d), 3.43 (3H, s), 3.60 (1H, d), 4.12 (3H, s), 7.27 (1H, d), 8.89 (1H, s) Step 2. 3-(4-Methoxycyclohex-1-en-1-yl)-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine A mixture of Cs2CO3 (496.78 mg, 1.525 mmol, 2.50 equiv), Xantphos (70.58 mg, 0.122 mmol, 0.20 equiv), Pd(AcO)2 (27.39 mg, 0.122 mmol, 0.20 equiv), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (99.40 mg, 0.671 mmol, 1.10 equiv) and 5-chloro-3-(4-methoxycyclohex-1-en-1-yl)-1-methylpyrazolo[4,3-d]pyrimidine (170.00 mg, 0.610 mmol, 1.00 equiv) in dioxane (5.00 mL) was stirred for 3 h at 100 °C under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 12:1) to give 3-(4-methoxycyclohex-1-en-1-yl)-1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[4,3-d]pyrimidin-5-amine (100 mg, 41.99%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 392.4。 1 H NMR (300 MHz, DMSO-d6) δ 1.67 (1H, d), 2.00 (1H, s), 2.27 (1H, m), 2.46 (3H, d), 2.72 (1H, d), 3.17 (2H, d), 3.33 (3H, s), 3.53 (1H, s), 4.07 (3H, s), 7.08 (1H, s), 7.73 (1H, s), 8.39 (1H, s), 8.91 (1H, s), 9.19 (1H, s), 9.43 (1H, s) Step 3. 3-((1r,4r)-4-Methoxycyclohexyl)-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 7) and 3-((1s,4s)-4-methoxycyclohexyl)-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 8) A mixture of Pd / C (65.41 mg, 0.615 mmol, 3.00 equiv) and 3-(4-methoxycyclohex-1-en-1-yl)-1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[4,3-d]pyrimidin-5-amine (80.00 mg, 0.205 mmol, 1.00 equiv) in MeOH (150.00 mL) and THF (80.00 mL) was stirred overnight under a hydrogen atmosphere at room temperature. The resulting mixture was filtered and the filter cake was washed with MeOH (3×50 mL). The filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 12:1) to give the crude product. The crude product (80 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column 19*250 mm, 5um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 25 mL / min; gradient: 37B~41B, 7 min; RT1: 5.30 / 5.92) to give 3-((1r,4r)-4-methoxycyclohexyl)-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 7, 60 mg, 22.73%) as a white solid. LCMS: m / z (ESI), [M+H] + = 393.3. 1 H NMR (400 MHz, DMSO-d6) δ 1.28 (2H, q), 1.83 (2H, q), 2.09 (4H, t), 2.43 (3H, d), 2.95 (1H, t), 3.30 (4H, m), 4.04 (3H, s), 7.73 (1H, s), 8.38 (1H, s), 8.81 (1H, s), 9.16 (1H, s), 9.51 (1H, s); 3-((1s,4s)-4-Methoxycyclohexyl)-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 8, 10 mg, 12.19%) as a white solid. LCMS: m / z (ESI), [M+H] + = 393.2. 11H NMR (300 MHz, DMSO-d6) δ 1.57 (2H, t), 1.76 (2H, m), 1.86 (2H, m), 2.04 (2H, q), 2.43 (3H, d), 3.05 (1H, m), 3.20 (3H, s), 3.42 (1H, s), 4.04 (3H, s), 7.72 (1H, s), 8.37 (1H, s), 8.80 (1H, s), 9.14 (1H, s), 9.34 (1H, s). Example 9 Preparation of 3-(4,4-difluorocyclohexyl)-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 9)

[0192]

Chemical Structure

[0193] Step 1. 5-Chloro-3-(4,4-difluorocyclohex-1-en-1-yl)-1-methyl-1H-pyrazolo[4,3-d]pyrimidine A mixture of 5-chloro-3-iodo-1-methylpyrazolo[4,3-d]pyrimidine (200.00 mg, 0.679 mmol, 1.00 equivalent), 2-(4,4-difluorocyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (198.93 mg, 0.815 mmol, 1.20 equivalent), Pd(dppf)Cl2 (99.39 mg, 0.136 mmol, 0.20 equivalent) and K2CO3 (234.66 mg, 1.698 mmol, 2.50 equivalent) in dioxane (3.00 mL) and H2O (0.60 mL) was stirred for 2 hours at 80 °C under a nitrogen atmosphere. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (PE / EtOAc 2:3) to give 5-chloro-3-(4,4-difluorocyclohex-1-en-1-yl)-1-methylpyrazolo[4,3-d]pyrimidine (100 mg, 51.72%) as a yellow solid. LCMS: m / z (ESI), [M+H] += 285.3.

[0194] Step 2. 3-(4,4-Difluorocyclohex-1-en-1-yl)-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine A mixture of 5-chloro-3-(4,4-difluorocyclohex-1-en-1-yl)-1-methylpyrazolo[4,3-d]pyrimidine (100.00 mg, 0.351 mmol, 1.00 equiv), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (62.45 mg, 0.421 mmol, 1.20 equiv), Pd(OAc)2 (15.77 mg, 0.070 mmol, 0.20 equiv), Xantphos (60.97 mg, 0.105 mmol, 0.30 equiv) and Cs2CO3 (286.12 mg, 0.878 mmol, 2.50 equiv) in dioxane (2.50 mL) was stirred under a nitrogen atmosphere at 100 °C for 2 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 12:1) to give 3-(4,4-difluorocyclohex-1-en-1-yl)-1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[4,3-d]pyrimidin-5-amine (110 mg, 79.00%) as a white solid. LCMS: m / z (ESI), [M+H] + = 397.3.

[0195] Step 3. 3-(4,4-Difluorocyclohexyl)-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 9) A mixture of 3-(4,4-difluorocyclohex-1-en-1-yl)-1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[4,3-d]pyrimidin-5-amine (130.00 mg, 0.328 mmol, 1.00 equiv) and Pd / C (174.50 mg, 1.640 mmol, 5.00 equiv) in MeOH (60.00 mL) and THF (30.00 mL) was stirred for 5 h under a hydrogen atmosphere at 30 °C. The resulting mixture was filtered and the filter cake was washed with CH2Cl2 (3 × 40 mL). The filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 12:1). The crude product (70 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column 30 × 150 mm, 5 um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 34B~48B, 7 min; RT1: 5.97) to give 3-(4,4-difluorocyclohexyl)-1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[4,3-d]pyrimidin-5-amine (17 mg, 13.01%) as a pink solid. LCMS: m / z (ESI), [M+H] + = 399.3. 1 H NMR (300 MHz, MeOD-d4) δ 1.80 - 2.22 (8H, m), 2.49 (3H, d), 3.22 (1H, s), 4.03 (3H, s), 7.58 (1H, t), 8.27 (1H, s), 8.98 (1H, s), 9.62 (1H, s) Example 10 Preparation of 1-(methyl-d3)-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-3-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 10)

[0196]

Chem.

[0197] Step 1. 5-Chloro-3-iodo-1-(methyl-d3)-1H-pyrazolo[4,3-d]pyrimidine A mixture of Cs2CO3 (1045.60 mg, 3.209 mmol, 3.00 equiv), CD3I (775.31 mg, 5.349 mmol, 50 equiv) and 5-chloro-3-iodo-1H-pyrazolo[4,3-d]pyrimidine (300.00 mg, 1.070 mmol, 1.00 equiv) in DMF (6.00 mL) was stirred for 1 h at 0 °C under a nitrogen atmosphere. The desired product could be detected by LCMS. The resulting mixture was diluted with water (50 mL). The resulting mixture was extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with Sat. brine (3 × 50 mL) and Na2S2O3 (3 × 50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give 5-chloro-3-iodo-1-(methyl-d3)-1H-pyrazolo[4,3-d]pyrimidine (260 mg, 81.70%) as a pink solid. LCMS: m / z (ESI), [M+H] + = 298.0. 1 H NMR (400 MHz, DMSO-d6) δ 9.45 (1H, s). Step 2. 5-Chloro-3-(3,6-dihydro-2H-pyran-4-yl)-1-(methyl-d3)-1H-pyrazolo[4,3-d]pyrimidine A mixture of K2CO3 (313.58 mg, 2.269 mmol, 2.50 equiv), Pd(dppf)Cl2CH2Cl2 (148.23 mg, 0.182 mmol, 0.20 equiv), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (228.79 mg, 1.089 mmol, 1.20 equiv) and 5-chloro-3-iodo-1-(methyl-d3)-1H-pyrazolo[4,3-d]pyrimidine (270.00 mg, 0.908 mmol, 1.00 equiv) in dioxane (5.00 mL) and water (1.00 mL) was stirred for 2 h at 80 °C under a nitrogen atmosphere. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (hexane / EtOAc 1:2) to give 5-chloro-3-(3,6-dihydro-2H-pyran-4-yl)-1-(methyl-d3)-1H-pyrazolo[4,3-d]pyrimidine (100 mg, 43.43%) as a pink solid. LCMS: m / z (ESI), [M+H] + = 254.2。 1 H NMR (400 MHz, DMSO-d6) δ 2.62 (2H, m), 3.86 (2H, t), 4.33 (2H, q), 7.18 (1H, t), 9.43 (1H, s) Step 3. 3-(3,6-Dihydro-2H-pyran-4-yl)-1-(methyl-d3)-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine A mixture of Cs2CO3 (385.28 mg, 1.182 mmol, 3.00 equiv), Xantphos (45.61 mg, 0.079 mmol, 0.20 equiv), Pd(OAc)2 (17.70 mg, 0.079 mmol, 0.20 equiv), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (70.08 mg, 0.473 mmol, 1.20 equiv) and 5-chloro-3-(3,6-dihydro-2H-pyran-4-yl)-1-(methyl-d3)-1H-pyrazolo[4,3-d]pyrimidine (100.00 mg, 0.394 mmol, 1.00 equiv) in dioxane (3.00 mL) was stirred for 2 h at 100 °C under a nitrogen atmosphere. The desired product could be detected by LCMS. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 12:1) to give 3-(3,6-dihydro-2H-pyran-4-yl)-1-(methyl-d3)-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (100 mg, 69.43%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 366.3. 1 H NMR (400 MHz, DMSO-d6) δ 2.44 (2H, d), 2.61 (3H, s), 3.17 (1H, d), 3.85 (2H, t), 4.26 (2H, q), 7.72 (1H, s), 8.38 (1H, s), 8.94 (1H, s), 9.20 (1H, s), 9.37 (1H, s). Step 4. 1-(Methyl-d3)-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-3-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (Example 10) A mixture of Pd / C (87.37 mg, 0.821 mmol, 3.00 equiv) and 3-(3,6-dihydro-2H-pyran-4-yl)-1-(methyl-d3)-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (100.00 mg, 0.274 mmol, 1.00 equiv) in MeOH (10.00 mL) and THF (10.00 mL) was stirred for 2 h under a hydrogen atmosphere at 35 °C. The desired product could be detected by LCMS. The resulting mixture was filtered and the filter cake was washed with MeOH (3 × 30 mL) and DCM (3 × 30 mL). The filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 12:1) to give the crude product. The crude product (40 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column 19*250 mm, 5 um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 15B~35B, 7 min; RT1: 6.4) to give 1-(methyl-d3)-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-3-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine (10 mg, 9.85%) as a white solid. LCMS: m / z (ESI), [M+H] + = 368.2. 1 H NMR (400 MHz, DMSO-d6) δ 1.94 (4H, d), 2.43 (3H, d), 3.24 (1H, d), 3.48 (2H, m), 3.93 (2H, m), 7.71 (1H, s), 8.37 (1H, s), 8.81 (1H, s), 9.15 (1H, s), 9.33 (1H, s). Example 11 / 12 Preparation of 1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-3-[(1s,4s)-4-(difluoromethoxy)cyclohexyl]pyrazolo[4,3-d]pyrimidin-5-amine (Example 11) and 1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-3-[(1r,4r)-4-(difluoromethoxy)cyclohexyl]pyrazolo[4,3-d]pyrimidin-5-amine (Example 12)

[0198]

Chemical formula

[0199] Step 1. 4-(5-Chloro-1-methyl-1H-pyrazolo[4,3-d]pyrimidin-3-yl)cyclohex-3-en-1-ol To a 40 mL vial were added 5-chloro-3-iodo-1-methylpyrazolo[4,3-d]pyrimidine (500.00 mg, 1.698 mmol, 1.00 equivalent), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-ol (570.78 mg, 2.547 mmol, 1.50 equivalents), Pd(dppf)Cl2 (248.47 mg, 0.340 mmol, 0.20 equivalent), K2CO3 (938.64 mg, 6.792 mmol, 4.00 equivalents), dioxane (10.00 mL) and H2O (2.00 mL) at room temperature. The mixture was then stirred at 80 °C under a nitrogen atmosphere for 3 hours. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (PE:EA = 1:4) to give 4-[5-chloro-1-methylpyrazolo[4,3-d]pyrimidin-3-yl]cyclohex-3-en-1-ol (253 mg, 56.29%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 265.2.

[0200] Step 2. 5-Chloro-3-(4-(difluoromethoxy)cyclohex-1-enyl)-1-methyl-1H-pyrazolo[4,3-d]pyrimidine To a 40 mL vial, 4-[5-chloro-1-methylpyrazolo[4,3-d]pyrimidin-3-yl]cyclohex-3-en-1-ol (253.00 mg, 0.956 mmol, 1.00 equiv), CuI (63.71 mg, 0.335 mmol, 0.35 equiv), MeCN (10.00 mL) were added at 50 °C, and 2,2-difluoro-2-(fluorosulfonyl)acetic acid (510.61 mg, 2.867 mmol, 3.00 equiv) was added dropwise to the mixture. The mixture was then stirred at 50 °C in an air atmosphere for 3 h. The resulting mixture was diluted with water (10 mL). The aqueous layer was extracted with DCM (3 × 30 mL). The resulting mixture was diluted with DCM (5 mL). The resulting mixture was filtered, and the filter cake was washed with DCM (10 mL * 3). The filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (PE / EtOAc 1:1) to give 5-chloro-3-[4-(difluoromethoxy)cyclohex-1-en-1-yl]-1-methylpyrazolo[4,3-d]pyrimidine (180 mg, 47.87%) as a yellow solid. LCMS: m / z (ESI), [M + H] + = 315.2。

[0201] Step 3. 3-(4-(Difluoromethoxy)cyclohex-1-enyl)-1-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-amine To a 40 mL vial were added 5-chloro-3-[4-(difluoromethoxy)cyclohex-1-en-1-yl]-1-methylpyrazolo[4,3-d]pyrimidine (180.00 mg, 0.572 mmol, 1.00 equiv), and 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (101.69 mg, 0.686 mmol, 1.20 equiv), Pd(OAc)2 (38.52 mg, 0.172 mmol, 0.30 equiv), Xantphos (99.28 mg, 0.172 mmol, 0.30 equiv), Cs2CO3 (559.05 mg, 1.716 mmol, 3.00 equiv) and dioxane (10.00 mL) at room temperature. The mixture was then stirred at 70 °C under a nitrogen atmosphere for 3 h. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (DCM:MeOH 15:1) to give 3-[4-(difluoromethoxy)cyclohex-1-en-1-yl]-1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[4,3-d]pyrimidin-5-amine (135 mg, 55.35%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 427.1. 1 H NMR (400 MHz, CDCl3) δ 1.97 - 2.23 (3H, m), 2.55 (4H, s), 2.68 - 2.87 (2H, m), 2.98 (1H, d), 4.11 (3H, s), 4.54 - 4.62 (1H,m), 6.35 (1H, t), 7.00 (1H, s), 7.61 (1H, s), 8.29 (1H,s), 8.82 (1H, s), 9.90 (1H, s) Step 4. Preparation of 1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-3-[(1s,4s)-4-(difluoromethoxy)cyclohexyl]pyrazolo[4,3-d]pyrimidin-5-amine (Example 11) and 1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-3-[(1r,4r)-4-(difluoromethoxy)cyclohexyl]pyrazolo[4,3-d]pyrimidin-5-amine (Example 12) To a stirred mixture of 3-[4-(difluoromethoxy)cyclohex-1-en-1-yl]-1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[4,3-d]pyrimidin-5-amine (135.00 mg, 0.317 mmol, 1.00 eq) in MeOH (20 mL) was added Pd / C (168.45 mg, 1.583 mmol, 5.00 eq) under an air atmosphere. The resulting mixture was stirred under a hydrogen atmosphere at 40 °C for 4 h. The resulting mixture was filtered and the filter cake was washed with DCM (8 × 100 mL). The filtrate was concentrated under reduced pressure to afford a crude solid. The crude product was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column 30 × 150 mm, 5 um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 34B~54B, 7 min; RT1: 5.93) to give a solid. The product was then purified by Prep-HPLC under the following conditions (column: CHIRALPAK IG, 2*25 cm, 5 um; mobile phase A: Hex:DCM = 3:1, mobile phase B: EtOH; flow rate: 20 mL / min; gradient: 10B~10B, 15 min; RT1: 10; RT2: 11;) to give 1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-3-[(1s,4s)-4-(difluoromethoxy)cyclohexyl]pyrazolo[4,3-d]pyrimidin-5-amine (Example 11, 15 mg, 50.00%) as a white solid and 1-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-3-[(1r,4r)-4-(difluoromethoxy)cyclohexyl]pyrazolo[4,3-d]pyrimidin-5-amine (Example 12, 5 mg, 16.67%) as a white solid.

[0202] (Example 11) LCMS: m / z (ESI), [M+H] + = 429.3. 11H NMR (300 MHz, DMSO-d6) δ1.87 (2H, d), 1.97 (4H, d), 2.09 (2H, d), 2.43 (3H, s), 3.07 (1H, d), 4.04 (3H, s), 4.37 (1H,s), 6.69 (1H, t), 7.70 (1H, s), 8.36 (1H, s), 8.79 (1H,s), 9.14 (1H, s), 9.33 (1H, s); (Example 12) LCMS: m / z (ESI), [M + H] + = 429.3。 1 1H NMR (300 MHz, MeOD-d4) δ1.59 - 1.71 (2H, m), 1.95 - 2.09 (2H, m), 2.15 - 2.20 (4H, m), 2.55 (3H,s), 3.05 - 3.14 (1H, m), 4.09 (3H, s), 4.20 - 4.27 (1H, m), 6.71 (1H, t), 7.64 (1H, s), 8.31 (1H,s), 9.03 (1H, s), 9.85 (1H, s). Example 13 Preparation of 3 - methyl - N-(7 - methyl - [1,2,4]triazolo[1,5 - a]pyridin - 6 - yl)-1-(tetrahydro - 2H - pyran - 4 - yl)-1H - pyrazolo[3,4 - d]pyrimidin - 6 - amine (Example 13)

[0203]

Chemical Structure

[0204] Step 1. 6 - chloro - 3 - methyl - 1-(tetrahydro - 2H - pyran - 4 - yl)-1H - pyrazolo[3,4 - d]pyrimidine A stirred mixture of 1-(2,4-dichloropyrimidin-5-yl)ethanone (500.00 mg, 2.618 mmol, 1.00 equiv) and DIPEA (1353.26 mg, 10.471 mmol, 4.00 equiv) in THF (5.00 mL) was added oxan-4-ylhydrazine (364.89 mg, 3.141 mmol, 1.20 equiv) portionwise under a nitrogen atmosphere at room temperature. And the mixture was stirred for 2 h under a nitrogen atmosphere at room temperature. The resulting mixture was extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with water (3 × 10 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (PE / EtOAc 2:1) to give 6-chloro-3-methyl-1-(oxan-4-yl)pyrazolo[3,4-d]pyrimidine (350 mg, 52.91%) as a yellow solid. LCMS: m / z (ESI), [M+H] + =253.2 Step 2. 3-Methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 13) To a stirred mixture of 6-chloro-3-methyl-1-(oxan-4-yl)-1H-pyrazolo[3,4-d]pyrimidine (200 mg, 0.791 mmol, 1.00 equiv), Cs2CO3 (644.68 mg, 1.979 mmol, 2.50 equiv) and 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (140.72 mg, 0.950 mmol, 1.20 equiv) in dioxane (20 mL), BrettPhos Pd G3 (143.49 mg, 0.158 mmol, 0.20 equiv) was added portionwise under a nitrogen atmosphere at room temperature. And the mixture was stirred for 3 h under a nitrogen atmosphere at 100 °C. The resulting mixture was extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with water (3 × 50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC under the following conditions (column: X select CS time OBD column 30 × 150 mm 5um n; mobile phase A: water (0.05% TFA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 18% B~29% B, 7 min; t R : 6.30 min) to give 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1-(oxan-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (74 mg, 25.66%) as a white solid. LCMS: m / z (ESI), [M+H] + = 365.3 1 H NMR (DMSO-d6, 300 MHz) δ 1.7 - 1.9 (2H, m), 2.0 - 2.2 (2H, m), 2.4 - 2.5 (6H, m), 3.4 (2H, td), 3.9 - 4.1 (2H, m), 4.5 - 4.7 (1H, m), 7.8 (1H, s), 8.6 (1H, s), 8.9 (1H, s), 9.3 (1H, s), 9.3 (1H, s) Example 14 Preparation of 3-methyl-N-(7-methylimidazo[1,2-a]pyridin-6-yl)-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 14)

[0205] [Chem.]

[0206] Step 1. 6-Chloro-3-methyl-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidine A solution of 1-(2,4-dichloropyrimidin-5-yl)ethanone (250.00 mg, 1.309 mmol, 1.00 equiv), oxan-4-ylhydrazine (182.45 mg, 1.571 mmol, 1.20 equiv), and DIPEA (338.32 mg, 2.618 mmol, 2.00 equiv) in THF (10.00 mL) was stirred for 2 h at 0 °C under a nitrogen atmosphere. The resulting mixture was extracted with EtOAc (20 mL × 3). The combined organic layers were washed with brine (10 mL × 3) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (PE / EtOAc 5:1) to give 6-chloro-3-methyl-1-(oxan-4-yl)pyrazolo[3,4-d]pyrimidine (230 mg, 69.54%) as a white solid. LCMS: m / z (ESI), [M+H] + = 253.2.

[0207] Step 2. 3-Methyl-N-(7-methylimidazo[1,2-a]pyridin-6-yl)-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 14) A mixture of 7-methylimidazo[1,2-a]pyridin-6-amine (139.78 mg, 0.950 mmol, 1.50 equiv) and 6-chloro-3-methyl-1-(oxan-4-yl)pyrazolo[3,4-d]pyrimidine (160 mg, 0.633 mmol, 1.00 equiv), BrettPhos Pd G3 (57.40 mg, 0.063 mmol, 0.10 equiv), and Cs2CO3 (412.59 mg, 1.266 mmol, 2.00 equiv) in dioxane (5.00 mL) was stirred for 3 h at 100 °C under a nitrogen atmosphere. The residue was purified by silica gel column chromatography, eluting with CH2Cl2 / MeOH = (10:1) to give the crude product. The crude product was purified by Prep-HPLC to give a crude solid. The crude product was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30 × 150 mm, 5 um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 17B~37B, 7 min; RT1: 6.75) to give 3-methyl-N-[7-methylimidazo[1,2-a]pyridin-6-yl]-1-(oxan-4-yl)pyrazolo[3,4-d]pyrimidin-6-amine (140 mg, 60.84%) as an off-white solid. LCMS: m / z (ESI), [M+H] + = 364.1. 1 H NMR (DMSO-d6, 300 MHz) δ1.79 (2H, d), 2.02 - 2.18 (2H, m), 2.23 (3H, d), 2.42 (3H, s), 3.44 (2H, dd), 3.94 (2H, dd), 4.62 (1H, dd), 7.42 (1H, s), 7.48 (1H, d), 7.84 (1H, s), 8.69 (1H, s), 8.85 (1H, s), 9.04 (1H, s) Example 15 Preparation of 7-methyl-N-(5-methyl-7-(tetrahydro-2H-pyran-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-[1,2,4]triazolo[1,5-a]pyridin-6-amine (Example 15)

[0208]

Chem.

[0209] Step 1. 2-Chloro-5-methyl-7-(tetrahydro-2H-pyran-4-yl)-7H-pyrrolo[2,3-d]pyrimidine A mixture of 2-chloro-5-methyl-7H-pyrrolo[2,3-d]pyrimidine (400.00 mg, 2.387 mmol, 1.00 equiv), 4-bromooxane (3.94 g, 23.874 mmol, 10.00 equiv) and K2CO3 (824.62 mg, 5.967 mmol, 2.50 equiv) in DMSO (50.00 mL, 12.922 mmol) was stirred overnight at 120 °C under a nitrogen atmosphere. The resulting mixture was diluted with water (150 mL). The resulting mixture was extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine (3 × 30 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 10:1). The crude product (70 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30 × 150 mm, 5 um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 28B~48B, 7 min; RT1: 5.80) to give 2-chloro-5-methyl-7-(oxan-4-yl)pyrrolo[2,3-d]pyrimidine (40 mg, 6.66%) as a yellow solid.

[0210] Step 2. 7-Methyl-N-(5-methyl-7-(tetrahydro-2H-pyran-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-[1,2,4]triazolo[1,5-a]pyridin-6-amine (Example 15) A mixture of 2-chloro-5-methyl-7-(oxan-4-yl)pyrrolo[2,3-d]pyrimidine (30.00 mg, 0.119 mmol, 1.00 equiv), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (21.19 mg, 0.143 mmol, 1.20 equiv), Pd(AcO)2 (5.35 mg, 0.024 mmol, 0.20 equiv), Xantphos (20.69 mg, 0.036 mmol, 0.30 equiv) and Cs2CO3 (97.08 mg, 0.298 mmol, 2.50 equiv) in dioxane (1.50 mL) was stirred under a nitrogen atmosphere at 100 °C for 2 h. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 12:1) to give the crude product. The crude product (40 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30×150 mm 5um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 26B~46B, 7 min; RT1: 6.37) to give 5-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-7-(oxan-4-yl)pyrrolo[2,3-d]pyrimidin-2-amine (6 mg, 13.85%) as a white solid. LCMS: m / z (ESI), [M+H] + = 364.3. 1 H NMR (300 MHz, MeOD-d4) δ1.79-1.91 (2H, m), 1.95-2.14 (2H, m), 2.24 (3H, d), 2.43 (3H, d), 3.47 (2H, td), 3.93-4.05 (2H, m), 4.55-4.72 (1H, m), 7.18 (1H, d), 7.58-7.74 (1H, m), 8.37 (1H, s), 8.62 (2H, d), 9.32 (1H, s) Example 16 Preparation of N-(7-chloro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-3-methyl-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 16)

[0211]

Chem.

[0212] Step 1. N-(7-Chloro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-3-methyl-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 16) A mixture of 7-chloro-[1,2,4]triazolo[1,5-a]pyridin-6-amine (50.00 mg, 0.297 mmol, 1.00 equiv), 6-chloro-3-methyl-1-(oxan-4-yl)pyrazolo[3,4-d]pyrimidine (74.95 mg, 0.297 mmol, 1.00 equiv), Pd(AcO)2 (13.32 mg, 0.059 mmol, 0.20 equiv), Xantphos (51.48 mg, 0.089 mmol, 0.30 equiv) and Cs2CO3 (241.59 mg, 0.741 mmol, 2.50 equiv) in dioxane (4.00 mL) was stirred under a nitrogen atmosphere at 100 °C for 2 h. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 12:1). The crude product (90 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 19*250 mm, 5 um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 25 mL / min; gradient: 30B~40B, 7 min; RT1: 6.62) to give N-[7-chloro-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-3-methyl-1-(oxan-4-yl)pyrazolo[3,4-d]pyrimidin-6-amine (32.77 mg, 28.71%) as a white solid. LCMS: m / z (ESI), [M+H] + = 385.2。 11H NMR (300 MHz, DMSO-d6) δ 1.84 (2H, d), 2.13 (2H, dd), 2.46 (3H, s), 3.39 - 3.57 (2H, m), 3.97 (2H, dd), 4.54 - 4.77 (1H, m), 8.23 (1H, s), 8.57 (1H, s), 8.93 (1H, s), 9.36 (2H, d). Example 17 Preparation of N-(7-Methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(tetrahydro-2H-pyran-4-yl)-3-(trifluoromethyl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 17)

[0213]

Chemical Structure

[0214] Step 1. 6-Chloro-1-(tetrahydro-2H-pyran-4-yl)-3-(trifluoromethyl)-1H-pyrazolo[3,4-d]pyrimidine To a stirred mixture of 6-chloro-3-(trifluoromethyl)-1H-pyrazolo[3,4-d]pyrimidine (340.00 mg, 1.528 mmol, 1.00 equivalent), oxan-4-ol (624.11 mg, 6.111 mmol, 4.00 equivalents) and PPh3 (1442.48 mg, 5.500 mmol, 3.60 equivalents) in THF (2.50 mL) was added DIAD (1112.07 mg, 5.500 mmol, 3.60 equivalents) dropwise under a nitrogen atmosphere at 0 °C. The resulting mixture was extracted with EtOAc (3 × 20 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 10:1) to give 6-chloro-1-(tetrahydro-2H-pyran-4-yl)-3-(trifluoromethyl)-1H-pyrazolo[3,4-d]pyrimidine (370 mg, 78.98%) as a white solid. LCMS: m / z (ESI), [M + H] + = 307.3.

[0215] Step 2. N-(7-Methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(tetrahydro-2H-pyran-4-yl)-3-(trifluoromethyl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 17) A mixture of 6-chloro-1-(oxan-4-yl)-3-(trifluoromethyl)pyrazolo[3,4-d]pyrimidine (70.00 mg, 0.228 mmol, 1.00 equiv), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (33.82 mg, 0.228 mmol, 1.00 equiv), Pd(AcO)2 (10.25 mg, 0.046 mmol, 0.20 equiv), Xantphos (39.62 mg, 0.068 mmol, 0.30 equiv) and Cs2CO3 (185.93 mg, 0.571 mmol, 2.50 equiv) in dioxane (3.00 mL) was stirred at 70 °C for 3 h under a nitrogen atmosphere. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 12:1). The crude product (90 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30×150 mm, 5um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 30B~50B, 7 min; RT1: 6.67) to give N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1-(oxan-4-yl)-3-(trifluoromethyl)pyrazolo[3,4-d]pyrimidin-6-amine (54.44 mg, 53.86%) as a white solid. LCMS: m / z (ESI), [M+H] + = 419.2。 1 H NMR (300 MHz, DMSO-d6) δ 1.77 - 2.01 (2H, m), 2.01 - 2.24 (2H, m), 2.39 (3H, d), 3.40 - 3.59 (2H, m), 3.98 (2H, d), 4.82 (1H, t), 7.78 (1H, d), 8.44 (1H, s), 9.13 (2H, d), 9.69 (1H, s) Example 18 Preparation of 1-(4-methoxybenzyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 18)

[0216]

Chemical formula

[0217] Step 1. 6-Chloro-1-(4-methoxybenzyl)-3-methyl-1H-pyrazolo[3,4-d]pyrimidine [(4-Methoxyphenyl)methyl]hydrazine (764.93 mg, 5.026 mmol, 1.50 equivalents) was added dropwise to a stirred mixture of 1-(2,4-dichloropyrimidin-5-yl)ethanone (640.00 mg, 3.351 mmol, 1.00 equivalent) and DIPEA (433.04 mg, 3.351 mmol, 1.00 equivalent) in THF under an air atmosphere at 0 °C. The resulting mixture was stirred for 2 hours under an air atmosphere at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (PE:EA 1:1) to give 6-chloro-1-[(4-methoxyphenyl)methyl]-3-methylpyrazolo[3,4-d]pyrimidine (623 mg, 64.40%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 289.2 1 H NMR (300 MHz, CDCl3) δ 2.58 (3H, s), 3.77 (3H, s), 5.47 (2H, s), 6.82 - 6.87 (2H, m), 7.30 - 7.35 (2H, m), 8.90 (1H, s) Step 2. 1-(4-methoxybenzyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 18) To a stirred mixture of 6-chloro-1-[(4-methoxyphenyl)methyl]-3-methylpyrazolo[3,4-d]pyrimidine (100.00 mg, 0.346 mmol, 1.00 equiv) and 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (76.97 mg, 0.519 mmol, 1.50 equiv) in dioxane (5 mL) were added Cs2CO3 (338.53 mg, 1.039 mmol, 3.00 equiv), Xantphos (40.08 mg, 0.069 mmol, 0.20 equiv), and Pd(AcO)2 (15.55 mg, 0.069 mmol, 0.20 equiv) dropwise under an air atmosphere at room temperature. The resulting mixture was stirred under a nitrogen atmosphere at 80 °C for 2 h. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 10:1) to give a yellow solid. The crude product (70 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 26B~46B, 7 min; RT1: 7.07) to give 1-[(4-methoxyphenyl)methyl]-3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[3,4-d]pyrimidin-6-amine (20 mg, 28.57%) as a white solid. LCMS: m / z (ESI), [M+H] + = 401.2。 1 H NMR (400 MHz, DMSO-d6) δ 2.38 (3H, s), 2.44 (3H, s), 3.71 (3H, s), 5.24 (2H, s), 6.85 - 6.88 (2H, m), 7.18 (2H, t), 7.77 (1H, s), 8.43 (1H, s), 8.95 (1H, s), 9.22 (2H, d) Example 20 Preparation of 1-cyclohexyl-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 20)

[0218] [Chem.]

[0219] Step 1. 6-Chloro-1-cyclohexyl-3-methyl-1H-pyrazolo[3,4-d]pyrimidine Cyclohexylhydrazine hydrochloride (184.56 mg, 1.225 mmol, 1.30 eq) was added portionwise to a stirred mixture of 1-(2,4-dichloropyrimidin-5-yl)ethanone (180.00 mg, 0.942 mmol, 1.00 eq) and DIPEA (487.17 mg, 3.769 mmol, 4.00 eq) in THF (10 mL) under an air atmosphere at 0 °C. The resulting mixture was stirred for 2 h under an air atmosphere at 25 °C. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (PE / EA 1:2) to give 6-chloro-1-cyclohexyl-3-methylpyrazolo[3,4-d]pyrimidine (145 mg, 61.37%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 251.2. 1 H NMR (300 MHz, CDCl3) δ 1.42 - 1.48 (2H, m), 1.51 - 1.55 (2H, m), 1.99 - 2.08 (6H, m), 2.60 (3H, s), 4.66 - 4.76 (1H, m), 8.90 (1H, s) Step 2. 1-Cyclohexyl-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 20) To a stirred mixture of 6-chloro-1-cyclohexyl-3-methylpyrazolo[3,4-d]pyrimidine (120.00 mg, 0.479 mmol, 1.00 equiv) and 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (106.37 mg, 0.718 mmol, 1.50 equiv) in dioxane (6 mL) were added Cs2CO3 (467.81 mg, 1.436 mmol, 3.00 equiv) and Xantphos (55.39 mg, 0.096 mmol, 0.20 equiv), Pd(AcO)2 (21.49 mg, 0.096 mmol, 0.20 equiv) portionwise under an air atmosphere at room temperature. The resulting mixture was stirred under a nitrogen atmosphere at 80 °C for 2 h. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 10:1) to give a crude solid. The crude product (80 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30*150mm 5um; phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: %B; 254; 220 nm; RT1: 6.50) to give 1-cyclohexyl-3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[3,4-d]pyrimidin-6-amine (60 mg, 34.59%) as a white solid. LCMS: m / z (ESI), [M+H] + = 363.2. 1 H NMR (300 MHz, DMSO-d6) δ 1.23 - 1.38 (3H, m), 1.67 (1H, d), 1.81 - 1.97 (6H, m), 2.41 - 2.44 (6H, m), 4.36 - 4.43 (1H, m), 7.74 (1H, s), 8.41 (1H, s), 8.91 (1H, s), 9.15 (1H, s), 9.22 (1H, s). Example 21 Preparation of 1-(4,4-difluorocyclohexyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 21)

[0220] [Chemical formula]

[0221] Step 1. 6-Chloro-1-(4,4-difluorocyclohexyl)-3-methyl-1H-pyrazolo[3,4-d]pyrimidine A mixture of DIPEA (338.32 mg, 2.618 mmol, 5.00 eq), (4,4-difluorocyclohexyl)hydrazine (86.48 mg, 0.576 mmol, 1.10 eq) and 1-(2,4-dichloropyrimidin-5-yl)ethanone (100.00 mg, 0.524 mmol, 1.00 eq) in THF (5.00 mL) was stirred for 3 h at 0 °C under a nitrogen atmosphere. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (hexane / EtOAc 1:1) to give 6-chloro-1-(4,4-difluorocyclohexyl)-3-methylpyrazolo[3,4-d]pyrimidine (80 mg, 53.30%) as a white solid. LCMS: m / z (ESI), [M+H] + = 287.3. 1 H NMR (300 MHz, CDCl3) δ 2.03 (4H, d), 2.35 (4H, d), 2.60 (3H, s), 4.85 (1H, t), 8.92 (1H, s). Step 2. 1-(4,4-Difluorocyclohexyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 21) A mixture of Cs2CO3 (227.28 mg, 0.698 mmol, 2.50 equiv), Xantphos (32.29 mg, 0.056 mmol, 0.20 equiv), Pd(AcO)2 (12.53 mg, 0.056 mmol, 0.20 equiv), 7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-amine (45.48 mg, 0.307 mmol, 1.10 equiv) and 6-chloro-1-(4,4-difluorocyclohexyl)-3-methylpyrazolo[3,4-d]pyrimidine (80.00 mg, 0.279 mmol, 1.00 equiv) in dioxane (3.00 mL) was stirred for 2 h under a nitrogen atmosphere at 100 °C. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 12:1) to give the crude product. The crude product (100 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 19*250 mm, 5 um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 31B~51B, 7 min; RT1: 6.30) to give 1-(4,4-difluorocyclohexyl)-3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[3,4-d]pyrimidin-6-amine (80 mg, 71.96%) as a white solid. LCMS: m / z (ESI), [M+H] + = 399.3. 1 H NMR (300 MHz, DMSO-d6) δ 1.99 (3H, s), 2.15 (5H, s), 2.38 (3H, d), 2.45 (3H, s), 4.62 (1H, s), 7.75 (1H, d), 8.42 (1H, s), 8.93 (1H, s), 9.13 (1H, s), 9.22 (1H, s). Example 22 Preparation of 1-(4,4-Difluorocyclohexyl)-3-methyl-N-(7-methylimidazo[1,2-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 22)

[0222]

Chemical Structure

[0223] Step 1. 1-(4,4-Difluorocyclohexyl)-3-methyl-N-(7-methylimidazo[1,2-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 22) A mixture of 6-chloro-1-(4,4-difluorocyclohexyl)-3-methylpyrazolo[3,4-d]pyrimidine (67.00 mg, 0.234 mmol, 1.00 equiv), 7-methylimidazo[1,2-a]pyridin-6-amine (68.79 mg, 0.467 mmol, 2.00 equiv), Pd(AcO)2 (10.49 mg, 0.047 mmol, 0.20 equiv), Xantphos (40.56 mg, 0.070 mmol, 0.30 equiv) and Cs2CO3 (190.35 mg, 0.584 mmol, 2.50 equiv) in dioxane (5.00 mL) was stirred for 2 h at 80 °C under a nitrogen atmosphere. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 12:1). The crude product (50 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30×150 mm, 5 um; mobile phase A: water (10 mmol / L NH4HCO3 + 0.1% NH3.H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 38B~50B, 7 min; RT1: 5.63) to give 1-(4,4-difluorocyclohexyl)-3-methyl-N-[7-methylimidazo[1,2-a]pyridin-6-yl]pyrazolo[3,4-d]pyrimidin-6-amine (20 mg) as a white solid. LCMS: m / z (ESI), [M+H] + = 398.2。 1 H NMR (300 MHz, DMSO-d6) δ 1.95 (3H, d), 2.15 (5H, d), 2.26 (3H, d), 2.44 (3H, s), 4.62 (1H, s), 7.44 (1H, s), 7.50 (1H, d), 7.86 (1H, t), 8.72 (1H, s), 8.88 (1H, s), 9.02 (1H, s) Example 26 Preparation of 1-((1r,4r)-4-methoxycyclohexyl)-3-methyl-N-(7-methylimidazo[1,2-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 26)

[0224]

Chemical Structure

[0225] Step 1. 1-(4-Methoxybenzyl)-3-methyl-N-(7-methylimidazo[1,2-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine A mixture of 6-chloro-1-[(4-methoxyphenyl)methyl]-3-methylpyrazolo[3,4-d]pyrimidine (200.00 mg, 0.693 mmol, 1.00 equivalent), 7-methylimidazo[1,2-a]pyridin-6-amine (152.92 mg, 1.039 mmol, 1.50 equivalents), Xantphos (120.24 mg, 0.208 mmol, 0.30 equivalent), Pd(AcO)2 (31.10 mg, 0.139 mmol, 0.20 equivalent) and Cs2CO3 (564.21 mg, 1.732 mmol, 2.50 equivalents) in dioxane (3 mL) was stirred for 2 hours under a nitrogen atmosphere at 80 °C. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 12:1) to give 1-[(4-methoxyphenyl)methyl]-3-methyl-N-[7-methylimidazo[1,2-a]pyridin-6-yl]pyrazolo[3,4-d]pyrimidin-6-amine (200 mg, 72.28%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 400.3.

[0226] Step 2. 3-Methyl-N-(7-methylimidazo[1,2-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine A solution of 1-[(4-methoxyphenyl)methyl]-3-methyl-N-[7-methylimidazo[1,2-a]pyridin-6-yl]pyrazolo[3,4-d]pyrimidin-6-amine (190.00 mg, 0.476 mmol, 1.00 equiv) and TFA (70.00 mL, 613.910 mmol, 1981.34 equiv) was stirred for 2 days at 80 °C under an air atmosphere. The resulting mixture was concentrated under vacuum. The resulting mixture was diluted with DCM (10 mL). The mixture was adjusted to pH 8 with saturated NaHCO3 (aqueous solution) by filtration and washed with DCM (2 × 3 mL). The precipitated solid was collected to give 3-methyl-N-[7-methylimidazo[1,2-a]pyridin-6-yl]-1H-pyrazolo[3,4-d]pyrimidin-6-amine (160 mg, 80.00%). LCMS: m / z (ESI), [M+H] + = 280.2.

[0227] Step 3. 1-((1r,4r)-4-Methoxycyclohexyl)-3-methyl-N-(7-methylimidazo[1,2-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 26) A mixture of 3-methyl-N-[7-methylimidazo[1,2-a]pyridin-6-yl]-1H-pyrazolo[3,4-d]pyrimidin-6-amine (150.00 mg, 0.537 mmol, 1.00 equiv), 4-methoxycyclohexan-1-ol (174.79 mg, 1.343 mmol, 2.50 equiv) and PPh3 (422.58 mg, 1.611 mmol, 3.00 equiv) in THF (2.50 mL) was stirred for 20 min at 0 °C under a nitrogen atmosphere, then DIAD (325.78 mg, 1.611 mmol, 3.00 equiv) was added and the mixture was stirred for 2 h at 70 °C. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / EtOAc = 12:1) to give a crude solid. The crude product (60.00 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30×150 mm, 5um; mobile phase A: water (10 mMOL / L NH4HCO3 + 0.1% NH3.H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 29B~39B, 9 min; RT1: 6.22,7.43) to give 3-methyl-N-[7-methylimidazo[1,2-a]pyridin-6-yl]-1-[(1r,4r)-4-methoxycyclohexyl]pyrazolo[3,4-d]pyrimidin-6-amine (16 mg, 26.67%) as a white solid. LCMS: m / z (ESI), [M+H] + = 392.2. 1 H NMR (300 MHz, MeOD-d4) δ 1.31 - 1.46 (2H, m), 2.00 (2H, s), 2.03 - 2.14 (2H, m), 2.21 (2H, d), 2.39 (3H, d), 2.50 (3H, s), 3.23 (1H, d), 3.38 (3H, s), 4.51 (1H, t), 7.46 (1H, s), 7.54 (1H, d), 7.80 (1H, s), 8.82 (2H, s). Example 27 Preparation of 1-((1s,4s)-4-fluorocyclohexyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 27) and 1-((1r,4r)-4-fluorocyclohexyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 28)

[0228]

Chemical formula

[0229] Step 1. 4-Fluorocyclohexan-1-ol NaBH4 (0.98 mg, 0.026 mmol, 3.00 equivalents) was added portionwise to a stirred mixture of 4-fluorocyclohexan-1-one (1.00 g, 8.611 mmol, 1.00 equivalent) and MeOH (100.00 mL) at 0 °C. The resulting mixture was stirred at room temperature for 16 h. The reaction was quenched with water at 0 °C. The resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with water (50 mL). The resulting mixture was extracted with CH2Cl2 (3 × 50 mL). The combined organic layers were washed with brine and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give 4-fluorocyclohexan-1-ol (800 mg, 78.63%) as a pale yellow solid. 1 HNMR (300 MHz, DMSO-d6) δ1.52 - 1.56 (5H, m), 1.88 - 1.90 (3H, m), 3.54 - 3.57 (1H, m), 4.55 - 4.58(1H, m), 4.70 - 4.72(1H, m) Step 2. 4-Fluorocyclohexylmethanesulfonate To a stirred mixture of 4-fluorocyclohexan-1-ol (400.00 mg, 3.385 mmol, 1.00 eq) and TEA (1027.74 mg, 10.156 mmol, 3.00 eq) in DCM (10.00 mL) was added methanesulfonyl chloride (581.66 mg, 5.078 mmol, 1.50 eq) dropwise at 0 °C. The resulting mixture was stirred at room temperature for 2 h. The resulting mixture was diluted with water (50 mL). The resulting mixture was extracted with CH2Cl2 (3 × 50 mL). The combined organic layers were washed with brine and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The resulting mixture was concentrated under reduced pressure to give 4-fluorocyclohexyl methanesulfonate (650 mg, 97.84%) as a pale yellow solid. The crude product was used directly in the next step without further purification. 1 1H NMR (300 MHz, DMSO-d6) δ 1.52 -1.90 (8H, m), 3.22 (3H, s), 4.60- 4.78(2H, m) Step 3. 1-((1s,4s)-4-Fluorocyclohexyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 27) and 1-((1r,4r)-4-fluorocyclohexyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 28) A mixture of 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1H-pyrazolo[3,4-d]pyrimidin-6-amine (150.00 mg, 0.535 mmol, 1.00 equiv), 4-fluorocyclohexylmethanesulfonate (1050.18 mg, 5.352 mmol, 10.00 equiv) and Cs2CO3 (523.09 mg, 1.605 mmol, 3.00 equiv) in DMF (20.00 mL) was stirred under a nitrogen atmosphere at 110 °C for 16 h. The resulting mixture was diluted with water (20 mL). The resulting mixture was extracted with EtOAc (3 × 20 mL). The combined organic layers were washed with brine and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product (100 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30 × 150 mm, 5 um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 39B~59B, 7 min; RT1: 6.4) to give the following: 1-((1s,4s)-4-Fluorocyclohexyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine as a white solid (Example 27, 6.8 mg, 3.34%). LCMS: m / z (ESI), [M+H] + = 381.2。 1 HNMR (300 MHz, DMSO-d6) δ 1.66 - 1.68 (1H, m), 1.75 (3H, d), 2.04 - 2.06(2H, m), 2.18 - 2.19 (2H, m), 2.38 (3H, d), 2.45 (3H, s), 4.50 - 4.53 (1H, m), 4.81 - 4.93 (1H, s), 7.73 (1H, s), 8.40 (1H, s), 8.91 (1H, s), 9.15 (2H, d); 1-((1r,4r)-4-Fluorocyclohexyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine as a white solid (Example 28, 18.8 mg, 9.23%) LCMS: m / z (ESI), [M+H] + = 381.2. 1 HNMR (400 MHz, DMSO-d6) δ 1.63 - 1.65(2H, m), 1.92 - 1.96 (4H, m), 2.00 - 2.04 (2H, m), 2.14 (3H, s), 2.41 (3H, s), 4.47 - 4.49 (1H, m), 4.61 - 4.65 (1H, m), 7.74 (1H, s), 8.41 (1H, s), 8.91 (1H, s), 9.19 (2H, d). Example 42 Preparation of 3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(2-oxaspiro[3.3]heptan-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 42)

[0230]

Chemical formula

[0231] Step 1. 3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(2-oxaspiro[3.3]heptan-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 42) 3-Methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1H-pyrazolo[3,4-d]pyrimidin-6-amine (70.00 mg, 0.250 mmol, 1.00 equiv), 2-oxaspiro[3.3]heptan-6-ol (57.01 mg, 0.499 mmol, 2.00 equiv), and PPh3 (196.51 mg, 0.749 mmol, 3.00 equiv) in THF (10 mL) were stirred, and DIAD (151.50 mg, 0.749 mmol, 3.00 equiv) was added portionwise at 0 °C under a N2 atmosphere. The resulting mixture was stirred for 2 h at 70 °C under a nitrogen atmosphere. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (DCM / MeOH = 15:1) to give a crude solid. The crude product (80 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3 + 0.1% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 26B~36B, 7 min; RT1: 5.88) to give 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1-[2-oxaspiro[3.3]heptan-6-yl]pyrazolo[3,4-d]pyrimidin-6-amine (40 mg, 50.00%) as a white solid. LCMS: m / z (ESI), [M+H] + = 337.3. 1 H NMR (300 MHz, DMSO-d6) δ 2.37 (3H, s), 2.44 (3H, s), 2.65 - 2.80 (4H, m), 4.45 (2H, s), 4.64 (2H, s), 4.84 - 4.89 (1H, m), 7.76 (1H, s), 8.42 (1H, s), 8.92 (1H, s), 9.13 (1H, s), 9.18 (1H, s). The following compounds in Table 2 below are synthesized by a similar method described in Example 42.

[0232]

Table 3-1

[0233]

Table 3-2

[0234]

Table 3-3

[0235]

Table 3-4

[0236]

Table 3-5

[0237]

Table 3-6

[0238] Example 37 3-Methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(spiro[2.5]octan-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 37)

[0239]

Chemical formula

[0240] Step 1. 3-Methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(spiro[2.5]octan-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 37) A mixture of TMAD (276.43 mg, 1.605 mmol, 3.00 equiv), n-Bu3P (324.81 mg, 1.605 mmol, 3.00 equiv), spiro[2.5]octan-6-ol (202.61 mg, 1.605 mmol, 3.00 equiv), and 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1H-pyrazolo[3,4-d]pyrimidin-6-amine (150.00 mg, 0.535 mmol, 1.00 equiv) in THF (25.00 mL) was stirred for 2 h at 70 °C under a nitrogen atmosphere. The resulting mixture was diluted with water (100 mL). The resulting mixture was extracted with CH2Cl2 (3 × 50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was recrystallized from MeOH (20 mL) to give the crude product. The crude product (60 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 19*250 mm, 5 um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: MeOH; flow rate: 25 mL / min; gradient 58B~70B, 7 min) to give 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1-[spiro[2.5]octan-6-yl]pyrazolo[3,4-d]pyrimidin-6-amine (30 mg, 14.29%) as a white solid. LCMS: m / z (ESI), [M+H] + = 389.3。 1 H NMR (400 MHz, DMSO-d6) δ 0.20 (2H, d), 0.33 (2H, d), 0.98 (2H, d), 1.84 (4H, m), 2.08 (2H, m), 2.39 (3H, d), 2.45 (3H, s), 4.45 (1H, d), 7.74 (1H, s), 8.40 (1H, s), 8.91 (1H, s), 9.16 (2H, d) The following compounds in Table 3 below are synthesized by a similar method as described in Example 37.

[0241]

Table 4

[0242] Example 43 Preparation of 3-Methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(spiro[3.3]heptan-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 43)

[0243]

Chemical formula

[0244] Step 1. Spiro[3.3]heptan-2-ylmethanesulfonate Methanesulfonyl chloride (459.50 mg, 4.012 mmol, 1.50 equivalents) was added dropwise to a stirred mixture of spiro[3.3]heptan-2-ol (300.00 mg, 2.674 mmol, 1.00 equivalent) and TEA (811.89 mg, 8.023 mmol, 3.00 equivalents) in DCM (50.00 mL) under a nitrogen atmosphere at 0 °C. The resulting mixture was diluted with water (20 mL). The resulting mixture was extracted with CH2Cl2 (3 × 20 mL). The combined organic layers were washed with brine and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. As a result, spiro[3.3]heptan-2-ylmethanesulfonate (500 mg, 98.26%) was obtained as a pale yellow oil.

[0245] Step 2. 3-Methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(spiro[3.3]heptan-2-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 43) A mixture of 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1H-pyrazolo[3,4-d]pyrimidin-6-amine (100.00 mg, 0.357 mmol, 1.00 equiv), spiro[3.3]heptan-2-ylmethanesulfonate (678.78 mg, 3.568 mmol, 10.00 equiv) and Cs2CO3 (348.73 mg, 1.070 mmol, 3.00 equiv) in DMF (20.00 mL) was stirred under a nitrogen atmosphere at 100 °C for 16 h. The resulting mixture was diluted with water (40 mL). The resulting mixture was extracted with EtOAc (3 × 100 mL). The combined organic layers were washed with brine and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 39B~59B, 7 min; RT1: 6.4) to give 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1-[spiro[3.3]heptan-2-yl]pyrazolo[3,4-d]pyrimidin-6-amine (31.8 mg, 23.80%) as a white solid. LCMS: m / z (ESI), [M+H] + = 375.3. 1 HNMR (400 MHz, DMSO-d6) δ 1.81 - 1.95 (2H, m), 1.89 - 1.91 (2H, m), 2.09 - 2.11 (2H, m), 2.42 (8H, d), 2.59 - 2.63 (2H, m), 4.89 - 4.93(1H, m), 7.76 (1H, s), 8.42 (1H, s), 8.92 (1H, s), 9.17 (2H, d) Example 44 Preparation of 3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(oxetan-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 44)

[0246] [Chemical]

[0247] Step 1. 3-Methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(oxetan-3-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 44) To a stirred mixture of 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1H-pyrazolo[3,4-d]pyrimidin-6-amine (80.00 mg, 0.285 mmol, 1.00 equiv) and 3-iodooxetane (78.76 mg, 0.428 mmol, 1.50 equiv) in DMF (10 mL) was added K2CO3 (118.34 mg, 0.856 mmol, 3.00 equiv) under an air atmosphere at room temperature. The resulting mixture was stirred for 2 h under an air atmosphere at 80 °C. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2:MeOH 12:1) to give the crude product. The crude product (100 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30×150 mm 5um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 12B~32B, 7 min; RT1: 6.62) to give 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1-(oxetan-3-yl)pyrazolo[3,4-d]pyrimidin-6-amine (Example 44, 40 mg, 41.67%) as a white solid. LCMS: m / z (ESI), [M+H] + = 337.2. 1 H NMR (300 MHz, DMSO-d6) δ 2.38 (3H, s), 2.49 (3H, s), 4.89 - 5.02 (4H, m), 5.72 (1H, t), 7.75 (1H, s), 8.42 (1H, s), 8.96 (1H, s), 9.15 (1H, s), 9.27 (1H, s) The following compounds in Table 4 below are synthesized by a similar method as described in Example 44.

[0248]

Table 5-1

[0249]

Table 5-2

[0250]

Table 5-3

[0251] Example 59 Preparation of 1-Isopropyl-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 59)

[0252]

Chemical formula

[0253] Step 1. 6-Chloro-1-isopropyl-3-methyl-1H-pyrazolo[3,4-d]pyrimidine Isopropylhydrazine (15.26 mg, 0.136 mmol, 1.30 equivalents) was added dropwise to a stirred mixture of 1-(2,4-dichloropyrimidin-5-yl)ethanone (150.00 mg, 0.785 mmol, 1.00 equivalent) and DIPEA (54.13 mg, 0.419 mmol, 4.00 equivalents) in THF under an air atmosphere at 0 °C. The resulting mixture was stirred for 2 hours under an air atmosphere at room temperature. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (PE:EA 1:1) to give 6-chloro-1-isopropyl-3-methylpyrazolo[3,4-d]pyrimidine (120 mg, 72.53%) as a yellow solid. LCMS: m / z (ESI), [M+H] + = 211.2. 11H NMR (300 MHz, CDCl3) δ 1.54 (6H, d), 2.61 (3H, s), 5.12 (1H, dd), 8.90 (1H, s) Step 2. 1-Isopropyl-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 59) To a stirred mixture of 6-chloro-1-isopropyl-3-methylpyrazolo[3,4-d]pyrimidine (120.00 mg, 0.570 mmol, 1.00 eq) and 7-methylimidazo[1,2-a]pyridin-6-amine (108.99 mg, 0.740 mmol, 1.30 eq) in dioxane (10 mL) were added Xantphos (65.92 mg, 0.114 mmol, 0.20 eq), Pd(AcO)2 (25.58 mg, 0.114 mmol, 0.20 eq) and Cs2CO3 (556.77 mg, 1.709 mmol, 3.00 eq) under an air atmosphere at room temperature. The resulting mixture was stirred for 2 h under a nitrogen atmosphere at 60 °C. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 10:1) to give the crude product. The crude product (100 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30 × 150 mm, 5 μm; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; RT1: 6.67) to give 1-isopropyl-3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[3,4-d]pyrimidin-6-amine (62 mg, 33.76%) as a white solid. LCMS: m / z (ESI), [M+H] + = 323.4. 1 1H NMR (300 MHz, DMSO-d6) δ 1.41 (6H, d), 2.40 (3H, d), 2.45 (3H, s), 4.73 - 4.82 (1H, m), 7.74 (1H, s), 8.41(1H, s), 8.92 (1H, s), 9.16 (1H, s), 9.19 (1H, s) Example 60 Preparation of 1-Isopropyl-3-methyl-N-(7-methylimidazo[1,2-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 60)

[0254]

Chemical formula

[0255] Step 1. 1-Isopropyl-3-methyl-N-(7-methylimidazo[1,2-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 60) A mixture of 6-chloro-1-isopropyl-3-methylpyrazolo[3,4-d]pyrimidine (80.00 mg, 0.380 mmol, 1.00 equivalent), 7-methylimidazo[1,2-a]pyridin-6-amine (67.07 mg, 0.456 mmol, 1.20 equivalent), Pd(AcO)2 (17.05 mg, 0.076 mmol, 0.20 equivalent), Xantphos (65.92 mg, 0.114 mmol, 0.30 equivalent) and Cs2CO3 (309.32 mg, 0.949 mmol, 2.50 equivalent) in dioxane (10.00 mL) was stirred for 3 hours under a nitrogen atmosphere at 100 °C. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 10:1). The crude product (100 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30×150 mm, 5 μm; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 21B~41B, 7 minutes; RT1: 7.02) to obtain 1-isopropyl-3-methyl-N-[7-methylimidazo[1,2-a]pyridin-6-yl]pyrazolo[3,4-d]pyrimidin-6-amine (55.77 mg, 45.70%) as a white solid. LCMS: m / z (ESI), [M+H] + = 322.2 11H NMR (300 MHz, DMSO-d6) δ 1.38 (6H, d), 2.25 (3H, d), 2.44 (3H, s), 4.67-4.89 (1H, m), 7.47 (2H, dd), 7.87 (1H, t), 8.67 (1H, s), 8.86 (1H, s), 9.03 (1H, s) Example 72 Preparation of 3-(3-methyl-6-((7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)propanenitrile (Example 72)

[0256]

Chemical formula

[0257] Step 1. 3-(3-methyl-6-((7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)amino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)propanenitrile (Example 72) To a stirred mixture of 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1H-pyrazolo[3,4-d]pyrimidin-6-amine (70.00 mg, 0.250 mmol, 1.00 equiv) and 3-bromopropanenitrile (66.92 mg, 0.499 mmol, 2.00 equiv) in DMF (10 mL) was added K2CO3 (103.55 mg, 0.749 mmol, 3.00 equiv). The resulting mixture was stirred at 80 °C under an air atmosphere for 2 h. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (DCM:MeOH = 15:1) to afford a crude solid. The crude product was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30 × 150 mm, 5 um; mobile phase A: water (0.05% NH3·H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 8B~28B, 7 min; RT1: 7.70) to give 3-[3-methyl-6-([7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]amino)pyrazolo[3,4-d]pyrimidin-1-yl]propanenitrile (40 mg, 48.05%) as a white solid. LCMS: m / z (ESI), [M+H] + = 334.3. 1 H NMR (300 MHz, DMSO-d6) δ 2.39 (3H, s), 2.47 (3H, s), 3.08 (2H, t), 4.36 (2H, t), 7.75 (1H, s), 8.41 (1H, s), 8.97 (1H, s), 9.17 (1H, s), 9.29 (1H, s). The following compounds in Table 5 below are synthesized by a similar method as described in Example 72.

[0258]

Table 6

[0259] Example 73 Preparation of 4-(3-Methyl-6-(7-methylimidazo[1,2-a]pyridin-6-ylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)benzonitrile (Example 73)

[0260] [Chemical Formula]

[0261] Step 1. 4-(3-Methyl-6-(7-methylimidazo[1,2-a]pyridin-6-ylamino)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)benzonitrile (Example 73) A mixture of 7-methyl-N-[3-methyl-1H-pyrazolo[3,4-d]pyrimidin-6-yl]-[1,2,4]triazolo[1,5-a]pyridin-6-amine (150.00 mg, 0.535 mmol, 1.00 equivalent), benzonitrile, 4-fluoro-benzonitrile (97.57 mg, 0.806 mmol, 1.50 equivalents), benzonitrile (97.22 mg, 0.803 mmol, 1.50 equivalents) and K2CO3 (221.88 mg, 1.605 mmol, 3.00 equivalents) in DMF (50.00 mL) was stirred at 100 °C for 16 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH = 10:1) to give 4-[3-methyl-6-([7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]amino)pyrazolo[3,4-d]pyrimidin-1-yl]benzonitrile (40 mg, crude) as a pale yellow solid. The crude product (40 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Shield RP18 OBD column, 19*250 mm, 10 um; mobile phase A: water (10 mMOL / L NH4HCO3 + 0.1% NH3·H2O), mobile phase B: ACN; flow rate: 20 mL / min; gradient: 36B~46B, 7 min; RT1: 5.73) to give 4-[3-methyl-6-([7-methylimidazo[1,2-a]pyridin-6-yl]amino)pyrazolo[3,4-d]pyrimidin-1-yl]benzonitrile (8.8 mg, 4.31%) as a white solid. LCMS: m / z (ESI), [M+H]+ = 381.3 1 HNMR (300 MHz, CDCl3) δ 2.45 (3H, d), 2.62 (3H, s), 7.08 (1H, s), 7.54 (2H, s), 7.65 (1H, s), 7.75 (2H, d), 8.42 - 8.44 (2H, m), 8.85 (1H, s), 9.06 (1H, d). Example 89 Preparation of 6-Methoxy-4-methyl-N-[3-methyl-1-(oxan-4-yl)pyrazolo[3,4-d]pyrimidin-6-yl]pyridin-3-amine

[0262]

Chemical formula

[0263] Step 1. 6-Methoxy-4-methyl-N-[3-methyl-1-(oxan-4-yl)pyrazolo[3,4-d]pyrimidin-6-yl]pyridin-3-amine (Example 89) A mixture of 6-chloro-3-methyl-1-(oxan-4-yl)pyrazolo[3,4-d]pyrimidine (80.00 mg, 0.317 mmol, 1.00 equiv), 6-methoxy-4-methylpyridin-3-amine (52.49 mg, 0.380 mmol, 1.20 equiv), Pd(AcO)2 (14.22 mg, 0.063 mmol, 0.20 equiv), Xantphos (54.95 mg, 0.095 mmol, 0.30 equiv) and Cs2CO3 (257.87 mg, 0.791 mmol, 2.50 equiv) in dioxane (2.50 mL) was stirred under a nitrogen atmosphere at 80 °C for 2 h. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (CH2Cl2 / MeOH 15:1) to give a crude solid. The crude product (100 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Shield RP18 OBD column, 30*150mm, 5um; mobile phase A: water (0.05% NH3H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 25B~55B, 7 min; 254; 220 nm; RT1: 5.20) to give 6-methoxy-4-methyl-N-[3-methyl-1-(oxan-4-yl)pyrazolo[3,4-d]pyrimidin-6-yl]pyridin-3-amine (70 mg, 62.39%) as a white solid. LCMS: m / z (ESI), [M+H] + = 355.2. 1 1H-NMR (300 MHz, DMSO-d6) δ 1.79 (2H, d), 1.99 - 2.20 (5H, m), 2.42 (3H, s), 3.43 - 3.56 (2H, m), 3.83 (3H, s), 3.96 (2H, dd), 4.58 (1H, t), 6.74 (1H, s), 8.11 (1H, s), 8.82 (1H, s), 8.98 (1H, s). Example 106 Preparation of 1-((1R,3r,5S)-8-oxa-bicyclo[3.2.1]octan-3-yl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine

[0264] [Chemistry]

[0265] Step 1. 1-((1R,3r,5S)-8-oxa-bicyclo[3.2.1]octan-3-yl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine To a stirred mixture of 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1H-pyrazolo[3,4-d]pyrimidin-6-amine (100.00 mg, 0.357 mmol, 1.00 eq), (1R,3S,5S)-8-oxabicyclo[3.2.1]octan-3-ol (137.18 mg, 1.070 mmol, 3.00 eq) and PPh3 (280.72 mg, 1.070 mmol, 3.00 eq) in THF (10.00 mL) was added DIAD (216.42 mg, 1.070 mmol, 3.00 eq) dropwise under a nitrogen atmosphere at 0 °C. The resulting mixture was stirred for 2 h under a nitrogen atmosphere at 70 °C. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (CH2Cl2 / MeOH 12:1) to give a crude solid. The crude product was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30×150 mm, 5um; mobile phase A: water (0.05% NH3H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 37B~57B, 7 min) to give 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1-[(1R,3R,5S)-8-oxabicyclo[3.2.1]octan-3-yl]pyrazolo[3,4-d]pyrimidin-6-amine (Example 106, 50 mg, 50.00%) as a white solid. LCMS: m / z (ESI), [M+H] + = 391.3. 11H NMR (300 MHz, DMSO-d6) δ 1.74(4H, s), 2.22-2.45(10H, m), 4.35(2H, s), 4.63(1H, s), 7.75(1H, s), 8.41(1H, s), 8.93(1H, s), 9.17 (1H, s), 9.21(1H, s). Example 107 Preparation of 1-((1R,3s,5S)-8-oxa-bicyclo[3.2.1]octan-3-yl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine

[0266]

Chemical formula

[0267] Step 1. 1-((1R,3s,5S)-8-oxa-bicyclo[3.2.1]octan-3-yl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine 3-Methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1H-pyrazolo[3,4-d]pyrimidin-6-amine (80.00 mg, 0.285 mmol, 1.00 equiv), (1R,3R,5S)-8-oxabicyclo[3.2.1]octan-3-ol (109.75 mg, 0.856 mmol, 3.00 equiv) and PPh3 (224.58 mg, 0.856 mmol, 3.00 equiv) in THF (16.00 mL) were stirred and DIAD (173.14 mg, 0.856 mmol, 3.00 equiv) was added dropwise thereto under a nitrogen atmosphere at 0 °C. The resulting mixture was stirred for 2 h under a nitrogen atmosphere at 70 °C. The resulting mixture was concentrated under vacuum. The residue was purified by Prep-TLC (CH2Cl2 / MeOH 12:1) to give a crude solid. The crude product was purified by Prep-HPLC under the following conditions (column: XBridge Prep OBD C18 column, 30×150 mm, 5 um; mobile phase A: water (0.05% NH3H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 37B~50B, 7 min) to give 1-((1R,3s,5S)-8-oxa-bicyclo[3.2.1]octan-3-yl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 107, 45 mg, 45.00%) as a white solid. LCMS: m / z (ESI), [M+H] + = 391.4. 1 H NMR (300 MHz, DMSO-d6) δ 1.78-1.94(6H, m), 2.07-2.25(2H, m),2.42(3H.s), 2.45(3H,s), 4.44(2H, s), 4.88-4.96(1H, m), 7.73(1H, s), 8.41(1H, s), 8.92(1H, s), 9.18(1H, s), 9.29(1H, s). Example 108 / 109 / 110 / 111 1-(3-Methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 108, Isomer 1) / 1-(3-Methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 109, Isomer 2) / 1-(3-Methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 110, Isomer 3) / 1-(3-Methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 111, Isomer 4)

[0268]

Chem.

[0269] Step 1. Preparation of 1-(3-Methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (mixture of 108 / 109) and preparation of 1-(3-Methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (mixture of 110 / 111) A stirred mixture of 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1H-pyrazolo[3,4-d]pyrimidin-6-amine (220.00 mg, 0.785 mmol, 1.00 equiv), PPh3 (617.59 mg, 2.355 mmol, 3.00 equiv) and 3-methoxycyclopentan-1-ol (273.52 mg, 2.355 mmol, 3.00 equiv) in THF (20.00 mL) was added dropwise with DIAD (476.13 mg, 2.355 mmol, 3.00 equiv) in THF (3 mL) at 0 °C for 10 min. The resulting mixture was stirred for 2 h under a nitrogen atmosphere at 70 °C. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH 10:1) to give 1-(3-methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (150 mg) as a pale yellow solid. The crude product (150 mg) was purified by Prep-HPLC under the following conditions (column: YMC-Actus Triart C18, 30*250,5um; mobile phase A: water (0.05% NH3H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 34B~46B, 8.5 min) to give 1-(3-methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (mixture of Examples 108 / 109, 25 mg, 16.67%) as a white solid, LCMS: m / z (ESI), [M+H] + = 379.3, 1HNMR (400 MHz, DMSO-d6) δ 1.81 - 1.85 (2H, m), 1.98 - 2.02 (2H, m), 2.09 - 2.11 (1H, m), 2.38 - 2.41 (4H, m), 2.45 (3H, s), 3.18 (3H, s), 3.83 - 3.85 (1H, m), 4.85 - 4.89 (1H, m), 7.74 (1H, s), 8.40 (1H, s), 8.91 (1H, s), 9.14 (2H, s), and 1-(3-methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (a mixture of Examples 110 / 111, 80 mg, 53.33%) was obtained as a white solid. LCMS: m / z (ESI), [M+H] + = 379.3. 1 HNMR (400 MHz, DMSO-d6) δ 1.61 - 1.63 (1H, m), 1.82 - 1.98 (1H, m), 2.38 - 2.41 (4H, m), 2.39 (3H, s), 2.45 (3H, s), 3.18 (3H, s), 3.93 - 3.96 (1H, m), 5.00 - 5.06 (1H, m), 7.74 (1H, s), 8.40 (1H, s), 8.91 (1H, s), 9.15 (2H, d). Step 2. Preparation of 1-(3-methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 108, isomer 1) / 1-(3-methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 109, isomer 2) The mixture of Example 108 / 109 (25 mg) was purified by Chiral-Prep-HPLC under the following conditions (column: CHIRALPAK AD-H, 2.0 cm I.D. * 25 cm L; mobile phase A: Hex (8 mmol / L NH3.MeOH)--HPLC, mobile phase B: IPA--HPLC; flow rate: 40 mL / min; gradient: 20B~20B, 18 minutes) to obtain 1-(3-methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 108, 4.5 mg, 18.00%) (isomer 1) as a white solid. LCMS: m / z (ESI), [M+H] + = 379.3. 1 HNMR (400 MHz, DMSO-d6) δ1.81 - 1.85 (2H, m), 1.98 - 2.01 (2H, m), 2.06 - 2.09 (1H, m), 2.38 - 2.43 (4H, m), 2.45 (3H, s), 3.18 (3H, s), 3.80 - 3.85 (1H, m), 4.82 - 4.87 (1H, m), 7.74 (1H, s), 8.40 (1H, s), 8.91 (1H, s), 9.14 (2H, s) and 1-(3-methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 109, 3.8 mg, 12.00%) (isomer 2) was obtained as a white solid. LCMS: m / z (ESI), [M+H] + = 379.3. 1 HNMR (400 MHz, DMSO-d6) δ 1.81 - 1.83 (2H, m), 1.97 - 2.01 (2H, m), 2.10 - 2.12 (1H, m), 2.37 - 2.43 (4H, m), 2.45 (3H, s), 3.18 (3H, s), 3.81 - 3.85 (1H, m), 4.80 - 4.89 (1H, m), 7.73 (1H, s), 8.40 (1H, s), 8.91 (1H, s), 9.14 (2H, s). Step 4. 1-(3-Methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 110, Isomer 3) / 1-(3-Methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 111, Isomer 4) A mixture of Examples 110 / 111 (80 mg) was purified by Prep-HPLC under the following conditions (column: CHIRALPAK AD-H, 2.0 cm I.D. * 25 cm L; mobile phase A: Hex (8 mmol / L NH3.MeOH)--HPLC, mobile phase B: IPA--HPLC; flow rate: 40 mL / min; gradient: 30B~30B, 12 min) to obtain 1-(3-methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 110, 39.1 mg, 48.87%) (Isomer 3) as a white solid, LCMS: m / z (ESI), [M+H] + = 379.3. 1 HNMR (400 MHz, DMSO-d6) δ 1.67 - 1.69(1H, m), 1.93 - 1.99 (1H, m), 2.10 - 2.15 (4H, m), 2.39 (3H, s), 2.44 (3H, s), 3.16 (3H, s), 3.94 (1H, s), 5.00 - 5.08 (1H, m), 7.74 (1H, s), 8.40 (1H, s), 8.91 (1H, s), 9.16 (2H, d) and 1-(3-methoxycyclopentyl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 111, 35.2 mg) (Isomer 4) was obtained as a white solid. LCMS: m / z (ESI), [M+H] + = 379.3. 1HNMR (400 MHz, DMSO-d6) δ 1.65 - 1.69 (1H, m), 1.93 - 1.97 (1H, m), 2.09 - 2.14 (4H, m), 2.39 (3H, d), 2.44 (3H, s), 3.15 (3H, s), 3.93 - 3.96 (1H, m), 5.00 - 5.08 (1H, m), 7.74 (1H, s), 8.41 (1H, s), 8.91 (1H, s), 9.16 (2H, d). Examples 112 / 113 / 114 / 115.

[0270] Preparation of 3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(3-methyltetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 112, isomer 1) / 3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(3-methyltetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 113, isomer 2) / 3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(3-methyltetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 114, isomer 3) / 3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(3-methyltetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 115, isomer 4)

[0271] [Chemical formula]

[0272] Step 1. 3-Methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1-[3-methyloxan-4-yl]pyrazolo[3,4-d]pyrimidin-6-amine (mixture of Examples 112 / 113) and 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1-[3-methyloxan-4-yl]pyrazolo[3,4-d]pyrimidin-6-amine (mixture of Examples 114 / 115) A mixture of 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1H-pyrazolo[3,4-d]pyrimidin-6-amine (200.00 mg, 0.714 mmol, 1.00 eq), 3-methyloxan-4-ol (248.65 mg, 2.141 mmol, 3.00 eq) and PPh3 (561.45 mg, 2.141 mmol, 3.00 eq) in THF (10.00 mL) was stirred at 0 °C, and DIAD (432.85 mg, 2.141 mmol, 3.00 eq) was added dropwise at 70 °C under a nitrogen atmosphere and stirred for 2 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by Prep_TLC (CH2Cl2 / MeOH 15:1) to obtain a crude solid. The crude product (120 mg) was purified by Prep-HPLC under the following conditions (column: XBridge Shield RP18 OBD column, 30*150 mm, 5um; mobile phase A: water (0.05% NH3H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 18B~48B, 7 min) to obtain 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1-[3-methyloxan-4-yl]pyrazolo[3,4-d]pyrimidin-6-amine (mixture of Examples 112 / 113, 30 mg, 25.00%) as a white solid, LCMS: m / z (ESI), [M+H] + = 379.3. 1HNMR (400 MHz, DMSO-d6) δ 0.52 (3H, d), 1.74 (2H, d), 2.06 - 2.31 (1H, m), 2.36 (3H, d), 2.45 (3H, s), 3.07 (1H, t), 3.43 (1H, t), 3.82 - 4.00 (2H, m), 4.14 - 4.27 (1H, m), 7.69 - 7.76 (1H, m), 8.39 (1H, s), 8.91 (1H, s), 9.15 (2H, d) and 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1-[-3-methyloxan-4-yl]pyrazolo[3,4-d]pyrimidin-6-amine (mixture of Examples 114 / 115, 70 mg, 57.75%) was obtained as a white solid. LCMS: m / z (ESI), [M+H] + = 379.3. 1 HNMR (400 MHz, DMSO-d6) δ 0.70 (3H, d), 1.74 - 1.91 (1H, m), 2.23 (1H, d), 2.37 (4H, d), 2.44 (3H, s), 3.44 - 3.65 (2H, m), 3.72 (1H, dd), 4.05 (1H, dt), 4.79 (1H, dt), 7.72 (1H, s), 8.39 (1H, s), 8.93 (1H, s), 9.17 (2H, s) Step 2. 3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(3-methyltetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 112, isomer 1) and 3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(3-methyltetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 113, isomer 2) The crude product 3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(3-methyltetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (mixture of Examples 112 / 113, 30.00 mg, 0.079 mmol, 1.00 equivalent) was purified by Prep-CHIRAL-HPLC under the following conditions (column: CHIRALPAK IE-3, 4.6*50 mm 3um; mobile phase A: Hex(0.1% DEA):EtOH = 50:50) to obtain 3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(3-methyltetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 112, isomer 1, 12.22 mg, 40.73%) as a white solid, LCMS: m / z(ESI), [M+H] + = 379.3. 1 HNMR (400 MHz, DMSO-d6) δ 0.52 (3H, d), 1.74 (2H, d), 2.06 - 2.31 (1H, m), 2.36 (3H, d), 2.45 (3H, s), 3.07 (1H, t), 3.43 (1H, t), 3.82 - 4.00 (2H, m), 4.14 - 4.27 (1H, m), 7.69 - 7.76 (1H, m), 8.39 (1H, s), 8.91 (1H, s), 9.15 (2H, d) and 3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(3-methyltetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 113, isomer 2, 8.37 mg, 27.90%) was obtained as a white solid. LCMS: m / z(ESI), [M+H] + = 379.3. 1HNMR (400 MHz, DMSO-d6) δ 0.52 (3H, d), 1.74 (2H, d), 2.06 - 2.31 (1H, m), 2.36 (3H, d), 2.45 (3H, s), 3.07 (1H, t), 3.43 (1H, t), 3.82 - 4.00 (2H, m), 4.14 - 4.27 (1H, m), 7.69 - 7.76 (1H, m), 8.39 (1H, s), 8.91 (1H, s), 9.15 (2H, d) Step 3. 3-Methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(3-methyltetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 114, isomer 3) and 3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(3-methyltetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 115, isomer 4) The crude product 3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(3-methyltetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (mixture of Example 114 / 115, 70.00 mg, 0.185 mmol, 1.00 equivalent) was purified by Prep-CHIRAL-HPLC under the following conditions (column: CHIRALPAK IE-3, 4.6 * 50 mm 3um; mobile phase A: Hex(0.1% DEA):EtOH = 50:50) to obtain 3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(3-methyltetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 114, isomer 3, 32.98 mg, 47.11%) as a white solid, LCMS: m / z(ESI), [M + H] + = 379.3. 11H NMR (300 MHz, DMSO-d6) δ 0.70 (3H, d), 1.74 - 1.91 (1H, m), 2.23 (1H, d), 2.37 (4H, d), 2.44 (3H, s), 3.44 - 3.65 (2H, m), 3.72 (1H, d), 4.05 (1H, t), 4.79 (1H, t), 7.72 (1H, s), 8.39 (1H, s), 8.93 (1H, s), 9.17 (2H, s), and 3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1-(3-methyltetrahydro-2H-pyran-4-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 115, isomer 4, 33.45 mg, 41.39%) was obtained as a white solid. LCMS: m / z (ESI), [M+H] + = 379.3 1 1H NMR (300 MHz, DMSO-d6) δ 0.70 (3H, d), 1.74 - 1.91 (1H, m), 2.23 (1H, d), 2.37 (4H, d), 2.44 (3H, s), 3.44 - 3.65 (2H, m), 3.72 (1H, d), 4.05 (1H, t), 4.79 (1H, t), 7.72 (1H, s), 8.39 (1H, s), 8.93 (1H, s), 9.17 (2H, s) Example 116 / 117 Preparation of 1-(2,2-dimethyltetrahydro-2H-pyran-4-yl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 116, isomer 1) and 1-(2,2-dimethyltetrahydro-2H-pyran-4-yl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 117, isomer 2)

[0273]

Chemical formula

[0274] Step 1. 1-(2,2-Dimethyltetrahydro-2H-pyran-4-yl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine To a 250 mL round-bottom flask were added 2,2-dimethyloxan-4-ol (627.03 mg, 4.816 mmol, 3.00 equiv), 3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]-1H-pyrazolo[3,4-d]pyrimidin-6-amine (450.00 mg, 1.605 mmol, 1.00 equiv), and PPh3 (1263.26 mg, 4.816 mmol, 3.00 equiv) in THF (60.00 mL) at 0 °C. A solution of DIAD (973.90 mg, 4.816 mmol, 3.00 equiv) in THF (10 mL) was added dropwise to the above solution under N2 at 0 °C, and the mixture was stirred at rt for 3 min. The reaction mixture was stirred at 70 °C for 2 h. The resulting mixture was concentrated under reduced pressure. The resulting mixture was diluted with DCM (20 mL), filtered, and the filter cake was washed with DCM (2 × 5 mL). The filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (CH2Cl2 / MeOH 15:1) to give the crude product. The crude product (250 mg) was purified by Prep-HPLC under the following conditions (column: YMC-Actus Triart C18, 30*250,5um; mobile phase A: water (0.05% NH3H2O), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 37B~57B, 7 min) to give 1-[2,2-dimethyloxan-4-yl]-3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[3,4-d]pyrimidin-6-amine (170 mg, 26.98%) as a white solid. LCMS: m / z (ESI), [M+H] + = 393.2。

[0275] Step 2. 1-(2,2-Dimethyltetrahydro-2H-pyran-4-yl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 116, Isomer 1) / 1-(2,2-Dimethyltetrahydro-2H-pyran-4-yl)-3-methyl-N-(7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-1H-pyrazolo[3,4-d]pyrimidin-6-amine (Example 117, Isomer 2) The crude product (170 mg) was purified by Chiral-Prep-HPLC under the following conditions (column: CHIRALPAK-AD-H-UL001, 20 * 250 mm, 5 μm; mobile phase A: Hex (8 mmol / L NH3.MeOH)--HPLC, mobile phase B: IPA--HPLC; flow rate: 20 mL / min; gradient: 25B~25B, 15 min; RT1: 10.12; RT2: 11.691) to obtain rel-1-[(4R)-2,2-Dimethyloxan-4-yl]-3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[3,4-d]pyrimidin-6-amine (Example 116, Isomer 1) (70 mg, 41.18%) LCMS: m / z (ESI), [M+H] + = 393.3 1 H-NMR (400 MHz, DMSO-d6) δ 1.21 (6H, d), 1.80 (2H, dd), 1.89 (1H, t), 2.05 (1H, qd), 2.35 - 2.48 (6H, m), 3.66 - 3.81 (2H, m), 4.75 - 4.95 (1H, m), 7.74 (1H, d), 8.40 (1H, s), 8.92 (1H, s), 9.20 (2H, d); and rel-1-[(4R)-2,2-Dimethyloxan-4-yl]-3-methyl-N-[7-methyl-[1,2,4]triazolo[1,5-a]pyridin-6-yl]pyrazolo[3,4-d]pyrimidin-6-amine (Example 117, Isomer 2) (70 mg, 41.18%) was obtained as a white solid, LCMS: m / z (ESI), [M+H] + = 393.3 1H-NMR (400 MHz, DMSO-d6) δ1.21 (6H, d), 1.80 (2H, dd), 1.89 (1H, t), 2.05 (1H, qd), 2.35 - 2.48 (6H, m), 3.66 - 3.81 (2H, m), 4.75 - 4.95 (1H, m), 7.74 (1H, d), 8.40 (1H, s), 8.92 (1H, s), 9.20 (2H, d). Biological Example The representative compounds disclosed herein were characterized in one or more of the following biological assays.

[0276] Biological Example 1: Enzymatic Assay The inhibitory activity of the compounds against DNA-PK was determined by measuring a fluorescence-labeled peptide substrate converted to a phosphorylated product by TR-FRET. All assays were performed in black Greiner 384-well low-volume plates (Greiner) with a total reaction volume of 6 μL and a final DMSO concentration of 0.5% (v / v). Full-length human DNAPK protein, fluorescein-P53(SeR15) peptide substrate (fluorescein-EPPLSQEAFADLWKK), and LanthaScreen™ Tb-anti-phospho-p53[pSeR15] antibody kit were purchased from Thermo Fisher Scientific. First, the DNA-PK protein was incubated with the compound for 30 minutes at room temperature in reaction buffer (50 mM HEPES pH 7.5, 0.01% Brij-35, 10 mM MgCl2, 1 mM EGTA, 1 mM DTT, 10 μg / ml Calf Thymus DNA). The reaction was then initiated by the addition of ATP and the fluorescein-P53(SeR15) peptide substrate. The kinase reaction (10 μM ATP, 1.6 μM peptide substrate) was quenched after 60 minutes by the addition of 6 μl of stop buffer containing 20 mM EDTA and 4 nM Tb anti-phospho-p53[Serl5] antibody. The reaction was incubated for an additional 1 hour and the plate was read on a Spark 20M (Tecan). The data were analyzed to determine the concentration of the compound that produced 50% inhibition of each kinase (IC50 ) was calculated using a 4-parameter logistic fit by XLfit. The DNA-PK inhibitory activities of representative compounds are shown in Table 5 below. It can be seen that the compounds of the present disclosure exhibited potent DNA-PK inhibitory activities.

[0277]

Table 7-1

[0278]

Table 7-2

[0279] Biological Example 2: Metabolic stability assay (rat hepatocyte Clint) The viability of cryopreserved hepatocytes was determined using trypan blue, and the cell concentration was adjusted to 106 cells / mL with buffer. A 1 μM compound (in acetonitrile; 0.01% DMSO) was incubated with 250 μL of hepatocytes (one million cells / mL) in a 96-well deep well plate. The reaction was stopped by adding 20 μL of the reaction mixture to 3 volumes of cold acetonitrile at various time points (0, 0.5, 5, 15, 30, 45, 60, 80, 100, and 120 minutes), and centrifuged at 4 °C for 15 minutes. 40 μL of the supernatant was diluted to 200 μL with pure water and analyzed using LC-MS / MS.

[0280] In vitro hepatocyte clearance was evaluated based on the determination of the elimination half-life (T 1 / 2 ) of the disappearance of the compound from its initial concentration. The peak area ratio of each compound (test or control) to the IS was calculated. The drug elimination rate constant k (min -1 ), T 1 / 2 (min), and in vitro intrinsic clearance CL int (μL / min / E6) were calculated according to the following equations: k = - slope T 1 / 2 = 0.693 / k CL int = k / C hep Here, C hep (cells × μL -1 ) is the cell concentration in the incubation system.

[0281] The data are shown in Table 6 below. Metabolic stability assay (human microsomes Clint) 1 μM of the compound was incubated at 37 °C in 250 μL of buffer (100 mM phosphate buffer, pH 7.4) containing 1 mM NADpH solution with 1 mg / mL of microsomes (HLM pooled with 20 mg / ml protein cone). At various time points 0, 0.5, 5, 10, 15, 20 and 30 minutes in a fresh 96-well plate, 20 μL of the incubation mix was quenched with 5 volumes of cold acetonitrile. The quenched plate was centrifuged at 4000 rpm for 15 minutes. 40 μL of the supernatant was diluted to 200 μL with pure water and analyzed using LC-MS / MS.

[0282] The in vitro intrinsic clearance CL of the drug in microsomes int (μl / min / mg) was calculated in a manner similar to Clint in hepatocytes. The data are also shown in Table 6 below. MDCKII-MDR1-BCRP efflux assay The apical to basolateral (A to B) and basolateral to apical (B to A) transport of the compound in HBSS (25 mM HEPES, pH 7.4) was measured across MDCKII-MDR1-BCRP cell monolayers. Incubation was carried out at approximately 37 °C for 120 minutes and the functionality of the test system was confirmed using 5 μM digoxin as a positive control. The transport of 5 μM compound and the control compound was determined by quantifying the substrate concentration in the culture medium in both the donor compartment at the start of the incubation period and the donor and acceptor compartments at the end of the incubation period. Using the data, the apparent permeability (Papp) was calculated. All incubations were performed in triplicate and the integrity of the cell monolayers was confirmed using marker Lucifer Yellow.

[0283] The permeability coefficient Pexact (cm / s) was calculated using the following equation: Papp = (dCr / dt) × Vr / (A × C0) Pexact = -(Vd × Vr) / (Vd + Vr) / A / t × ln(1 - (Vd + Vr) × Cr / (Vd × Cd + Vr × Cr)) The Pexact ratio was calculated using the following equation: Pexact or Papp ratio = Pexact or Papp(+ inhibitor) / Pexact or Papp(- inhibitor) The efflux ratio was calculated using the following equation: Efflux ratio = Pexact or Papp(BA) / Pexact or Papp(AB) Where dCr / dt is the cumulative concentration of the compound in the receptor chamber as a function of time (μM / s); Vr is the volume of the solution in the receptor chamber (0.1 mL on the apical side, 0.3 mL on the basolateral side); A is the surface area of transport, i.e., 0.11 cm2 for the monolayer area; and C0 is the initial concentration in the donor chamber (μM). The data are shown in Table 6 below.

[0284]

Table 8-1

[0285]

Table 8-2

[0286]

Table 8-3

[0287] Biological Example 3: Blood-brain barrier permeability assay Kp,uu, which is the relationship between the unbound drug concentrations in the brain and plasma, is a key to predicting CNS activity and is considered a major parameter to be measured and optimized in drug discovery (Di L et al., Journal of Medicinal Chemistry

[2013] , 56:2~12).

[0288] In vitro plasma and brain binding assays were performed using equilibrium dialysis with a semipermeable membrane. 5 μM of the test compound was added to plasma and diluted brain homogenate (1:4 with DPBS, pH 7.4) (in triplicate), and dialyzed in a plate slowly rotating at 37 °C for 18 h against an equal volume of 150 μL of 100 mM PBS buffer (pH 7.4). At the end of incubation, 50 μL aliquots from the acceptor side and 5 μL from the donor chamber were taken. The 5 μL sample was further diluted with 45 μL of blank plasma or brain homogenate. The paired samples were matrix-matched with either buffer or blank plasma / brain homogenate, mixed for 2 min, and then precipitated with 150 μL of cold acetonitrile having 100 ng / mL tolbutamide as the internal standard. After centrifugation at 4000 rpm for 20 min, the supernatant was diluted with 0.1% aqueous formic acid and analyzed by LC / MS / MS (API 4000, Applied Biosystems, Foster City). The unbound fraction (fu) of the test compound in brain homogenate and diluted plasma was calculated by the ratio of the reaction on the buffer side to the reaction on the brain homogenate / plasma side, and the unbound fractions (fu,pl and fu,br) of the test compound in undiluted plasma and tissue were calculated from the fu measured in homogenate and plasma using the following equation: fu,bl(fu,br)=(1 / D) / [(1 / fu - 1)+1 / D)]. D is the dilution factor.

[0289] The immediate oral absorption (SOA) model is an in vivo screening model for identifying the brain permeability of compounds. Six male Han Wistar rats purchased from Beijing Vital River were orally administered the compound at 10 mg / kg in 1% methylcellulose. Cerebrospinal fluid (CSF) was collected from the cisterna magna at 0.5, 1, 2, 4, 7, and 16 hours after dosing. Plasma samples were to be processed for plasma by centrifugation at approximately 4°C and 3,000 g within 30 minutes of collection. Plasma samples were to be transferred to labeled tubes and stored at -80°C until analysis. Brain tissue was harvested and homogenized in 3 volumes of 100 mM phosphate buffered saline (pH 7.4). All samples were stored at approximately -70°C before LC / MS / MS analysis.

[0290] Standards were prepared by adding blank plasma, brain homogenate, and artificial CSF ranging from 0.5 to 500 ng / mL. The homogenized brain tissue was precipitated together with plasma samples by adding 3 volumes of cold acetonitrile containing internal standards (40 ng / mL dexamethasone and 40 ng / mL diclofenac), and 10 μL of CSF samples were precipitated with 100 μL of cold acetonitrile containing internal standards. After 2 minutes of vortexing and 5 minutes of centrifugation at 14,000 rpm, the supernatant was analyzed by LC / MS / MS (API 4000, Applied Biosystems, Foster City). Two sets of standard curves were run at the start and end of each batch from plasma sample analysis. For brain and CSF samples, one standard curve was analyzed using test samples.

[0291] The total brain level expressed as the brain / plasma ratio (Kp) was measured by AUC brain / AUC plasma in rodents after oral administration. The free fraction of the test compound in biological matrices was determined by in vitro plasma and brain binding assays. Kp,uu was calculated by the following formula: Kp,uu = AUC(brain) / AUC(plasma) × (fu,brain / fu,plasma). The data are shown in Table 7 below.

[0292]

Table 9

[0293] Although the present disclosure has been particularly shown and described with reference to specific embodiments, some of which are preferred embodiments, various changes in form and detail may be made without departing from the spirit and scope of the disclosure as disclosed herein, as should be understood by those skilled in the art.

Claims

1. A compound of formula I 【Chemical Formula 1】 or a pharmaceutically acceptable salt thereof (wherein X １ , X ２ and X ３ are each independently C, CH, or N, provided that at least one of X １ , X ２ and X ３ is N, and at least one of X １ , X ２ and X ３ is C; The dashed line “—” means a bond between X １ and X ２ and between X ２ and X ３ and may be a single bond or a double bond, provided that at least one of the bonds between X １ and X ２ and between X ２ and X ３ is a single bond; R １ is C １～６ alkyl, where said C １～６ alkyl may be mono-substituted or independently multi-substituted by halogen or deuterium; R ２ is selected from C １～６ alkyl, C １～６ alkoxyl, —(CH ２ ) ｎ —Q, optionally these may be mono-substituted or independently multi-substituted by hydroxyl, cyano, halogen, C １～６ alkyl, C １～６ haloalkyl, C １～６ alkoxyl, C １～６ haloalkoxyl, where n is 0, 1 or 2, and Q is a 3- to 8-membered saturated carbocyclic or heterocyclic ring, phenyl, or a 6-membered unsaturated heterocyclic ring; R ３ is selected from halogen or C １～６ alkyl; R ４ is absent, or is cyano, C １～６ alkyl, C １～６ alkoxyl, -(CH ２ ) ｎ -Q, optionally one or more of which may be independently mono- or polysubstituted by C １～６ alkyl, where n is 0, 1 or 2 and Q is a 5-membered unsaturated heterocyclyl; Ring A is selected from the group consisting of the following [Chemical Formula 2] ().

2. The compound according to claim 1, having the structure of formula Ia [Chemical Formula 3] or a pharmaceutically acceptable salt thereof.

3. The compound according to claim 1, having the structure of formula Ib [Chemical Formula 4] or a pharmaceutically acceptable salt thereof.

4. The compound according to claim 1, having the structure of formula Ic [Chemical Formula 5] or a pharmaceutically acceptable salt thereof.

5. The compound according to claim 1, having the structure of formula Id [Chemical Formula 6] or a pharmaceutically acceptable salt thereof.

6. R ２is selected from methyl, ethyl, propyl, butyl, cyclopropyl, cyclobutyl, oxetanyl, cyclopentanyl, tetrahydrofuryl, cyclohexanyl, tetrahydropyranyl, cycloheptanyl, piperidinyl, phenyl, pyridinyl, pyridonyl, oxocanyl, dihydropyranyl, spiro[3.3]heptanyl, spiro[2.5]octanyl, bicyclo[1.1.1]pentanyl, bicyclo[3.2.1]octanyl, 8-oxabicyclo[3.2.1]octan-3-yl, and optionally these are mono- or independently polysubstituted by hydroxyl, cyano, halogen, C １～６ alkyl, C １～６ haloalkyl, C １～６ alkoxyl, C １～６ haloalkoxyl, and the compound according to claim 1 may be mono- or independently polysubstituted.

7. R ２ is 【Chemical Formula 7】 selected from, and optionally these are mono- or independently polysubstituted by hydroxyl, cyano, fluoro, chloro, bromo, methyl, ethyl, methoxyl, difluoromethyl, difluoromethoxyl or trifluoromethoxyl, and the compound according to claim 1.

8. R ２ is cyclohexanyl or tetrahydropyranyl, and optionally these are mono- or independently polysubstituted by halogen, C １～６ alkyl or C １～６ alkoxyl, and the compound according to claim 1.

9. R １ is methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl or isobutyl, and optionally these are mono- or independently polysubstituted by halogen or deuterium, and the compound according to claim 1.

10. R １The compound according to claim 1, wherein is methyl, ethyl, trifluoromethyl or trideuteriomethyl.

11. R ３ is C １～６ alkyl, the compound according to claim 1.

12. Ring A is 【Chemical Formula 8】 and R ３ is methyl, R ４ is either absent or selected from methyl, cyano, methoxyl, pyrazolyl, oxazolyl, where said pyrazolyl or oxazolyl may optionally be further mono - substituted or independently polysubstituted by C １～３ alkyl, the compound according to claim 1.

13. The compound according to claim 1 having the structure of formula Ie 【Chemical Formula 9】 or a pharmaceutically acceptable salt thereof (wherein X １ and X ３ one of is N and the other is C, and the dashed line “ - ” means the bond between X １ and N and between N and X ３ and may be a single bond or a double bond, provided that at least one of the bonds between X １ and N and between N and X ３ is a single bond; R １ is C １～３ alkyl, R ２ is cyclopentyl, cyclohexanyl, tetrahydropyranyl or 8 - oxabicyclo[3.2.1]octan - 3 - yl, and optionally these may be mono - substituted or independently polysubstituted by halogen or C １～３ alkoxyl, Y １ Y ２ and Y ３ are each independently C or N, provided that Y １ and Y ２ and Y ３ at least one of which is N; R ５ is halogen or C １～３ alkyl, R ６ is C １～３ alkyl).

14. R ２ is unsubstituted cyclopentyl, cyclohexanyl, tetrahydropyranyl or 8-oxabicyclo[3.2.1]octan-3-yl, optionally these may be mono-substituted or independently multi-substituted by halogen or C １～３ alkoxyl, the compound according to claim 13.

15. Y ３ is N, Y １ and Y ２ at least one of which is N, the compound according to claim 13.

16. R ５ is methyl, the compound according to claim 13.

17. 【Chemical Formula 10-1】 【Chemical Formula 10-2】 【Chemical Formula 10-3】 【Chemical Formula 10-4】 【Chemical Formula 10-5】 【Chemical Formula 10-6】 【Chemical Formula 10-7】 【Chemical Formula 10-8】 【Chemical Formula 10-9】 【Chemical Formula 10-10】 a compound selected from the group consisting of, or a pharmaceutically acceptable salt thereof.

18. A crystal of the compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof. Claim 19 A pharmaceutical composition comprising one or more compounds according to any one of claims 1 to 18 as a first active ingredient, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable diluent, excipient or carrier. Claim 20 A compound according to any one of claims 1 to 18, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 19, for inhibiting DNA-dependent protein kinase (DNA-PK). Claim 21 A pharmaceutical composition for inhibiting DNA-PK, comprising one or more compounds according to any one of claims 1 to 18 and a pharmaceutically acceptable salt thereof. Claim 22 A pharmaceutical composition for treating a DNA-PK-related disease in a subject, comprising an effective amount of one or more compounds according to any one of claims 1 to 18, or a pharmaceutically acceptable salt thereof. Claim 23 The pharmaceutical composition according to claim 22, wherein the subject is a warm-blooded animal such as a human. Claim 24 The pharmaceutical composition according to claim 22, wherein the DNA-PK-related disease is cancer. Claim 25 A pharmaceutical composition comprising a compound according to any one of claims 1 to 18, or a pharmaceutically acceptable salt thereof, for use in combination with a second therapeutic agent. Claim 26 The pharmaceutical composition according to claim 25, wherein the second therapeutic agent is an anti-cancer agent.

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