Novel imidazo[1,2-b]pyridazine-based compounds as cdks inhibitors and uses thereof

Abstract

This invention provides a novel compound, its use in inhibiting CDK12 and / or CDK13 activity and degrading cyclin K, a pharmaceutical composition comprising the compound, and its use in treating diseases (e.g., cancer) associated with CDK12 and / or CDK13 and / or cyclin K. According to the invention, these compounds exhibit excellent inhibitory activity and selectivity against CDK12 and / or CDK13, inducing the degradation of cyclin K, and therefore can be used for the treatment of cancers (e.g., breast cancer or gastric cancer).

Description

Technical Field

[0001] This invention relates to novel compounds that inhibit the activity of cyclin-dependent kinases (CDKs) (e.g., CDK12 and / or CDK13) and degrade cyclin K. Furthermore, this invention relates to pharmaceutical compositions comprising the compounds of this invention and their use in treating CDK-mediated diseases. Background Technology

[0002] One of the key phenomena in cancer is uncontrolled cell growth, which makes targeted therapies that inhibit cell division effective in treating cancer. Cyclins and cyclin-dependent kinases (CDKs) are a group of proteins that primarily control cell division (Mengna et al., CDK inhibitors in cancer therapy, an overview of recent development. 2021, Am J Cancer Res;11(5):1913-1935). Cyclin-dependent protein kinases (CDKs) are catalytic subunits of the serine / threonine protein kinase family. CDKs form complexes with cyclins and activate cyclins by phosphorylating serine or threonine residues in the matrix. Based on their cellular functions, CDKs can be divided into two categories: CDKs that control the cell cycle and CDKs that regulate cellular transcription.

[0003] The activity of CDKs is regulated through specific binding to cyclin regulatory units, such as cyclin A, cyclin B, cyclin C, cyclin D, and cyclin E. For example, in mammalian cells, CDK1, complexed with cyclin A / B, regulates the progression from G2 to M phase. CDK4 and CDK6, complexed with cyclin D, control the progression from G1 to S phase, which initiates DNA synthesis. CDK2, complexed with cyclin E / A, completes DNA synthesis in S phase. CDK7-9 form complexes with cyclins H, C, and T, respectively, to regulate gene transcription [Shigeaki et al., CDK1 inhibitor controls G2 / M phase transition and reverses DNA damage sensitivity. 2021, Biochem Biophys Res Commun. Apr 23;550:56-61], [Manuel et al., A CDK4 / 6-dependent phosphorylation gradient regulates the early to late G1 phase transition. 2021, Sci Rep. Jul 19;11(1):14736], [Stefan et al., Structural basis for CDK7 activation by MAT1 and Cyclin H. 2020, Proc NatlAcad Sci US A. Oct 27;117(43):26739-26748].

[0004] CDK12 and its ortholog CDK13 belong to the CDK family, which regulates transcription and post-transcriptional processes. Among the CDKs, mutations in CDK12 have been reported to be relatively high in various cancers. Furthermore, CDK12 and CDK13 form a complex with their cyclin chaperone (cyclin K) and phosphorylate the C-terminal domain of RNA polymerase II, which plays a key role in regulating kinase activity. In addition, CDK12 is known to regulate major signal transduction pathways associated with cancer induction and DNA damage responses, and inhibition of these pathways is expected to have anti-cancer effects. SR-4835 (international publication number WO2019 / 217421) is the most representative CDK12 / CDK13 inhibitor known to date, and its anti-cancer effects have been reported in numerous experiments (Victor et al., Therapeutic Targeting of CDK12 / CDK13 in Triple-Negative Breast Cancer. 2019, Cancer Cell. Nov 11;36(5):545-558.e7). However, there is a need in the field to develop CDK12 / 13 inhibitors with improved efficacy and selectivity compared to existing agents. Summary of the Invention

[0005] Technical issues

[0006] The object of this invention is to provide a compound of formula I, a stereoisomer thereof, hydrate thereof, solvate thereof, or pharmaceutically acceptable salt thereof, that inhibits the activity of CDKs (e.g., CDK12 and / or CDK13) and degrades cyclin K:

[0007] [Formula I]

[0008]

[0009] In equation I above,

[0010] R 1 The radical is halogenated, hydroxylated, cyano, C1-C6 alkoxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 ynyl, C3-C6 cycloalkyl, C3-C6 cycloalkyl-C1-C6 alkyl-, C3-C6 cycloalkyl-C2-C6 alkenyl-, or C3-C6 cycloalkyl-C2-C6 ynyl-, wherein R 1 Any C1-C6 alkoxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, and C3-C6 cycloalkyl groups may optionally be substituted with a halogen, hydroxyl, or cyano group;

[0011] R 2 For C6-C 10Aryl; a 5- or 6-membered heteroaryl group containing 1 to 2 nitrogen atoms; or a 4- to 12-membered heterocyclic group containing 1 nitrogen atom and linked via that nitrogen atom to an imidazopyridazine ring of Formula I, wherein the heterocyclic group may optionally contain an additional nitrogen atom or an oxygen atom, and

[0012] R 2 It may optionally be substituted by one or more substituents selected from the group consisting of:

[0013] (i) H, halogen, hydroxyl, cyano, C1-C6 alkyl, C3-C6 cycloalkyl,

[0014] (ii) C1-C6 alkyl groups substituted with halogen, hydroxyl, or cyano groups,

[0015] (iii) C3-C6 cycloalkyl groups substituted with halogen, hydroxyl, or cyano groups,

[0016] (iv) amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, (C3-C6 cycloalkyl)amino, (C1-C6 hydroxyalkyl)amino, (C1-C6 alkyl)carbonylamino, (C1-C6 hydroxyalkyl)carbonylamino, or (C1-C6 alkoxy)(C1-C6 alkyl)carbonylamino, and

[0017] (v) (C1-C6 alkyl)carbonyl, (C1-C6 hydroxyalkyl)carbonyl, aminocarbonyl, (C1-C6 alkyl)aminocarbonyl, di(C1-C6 alkyl)aminocarbonyl, carboxyl, or (C1-C6 alkoxy)carbonyl; and

[0018] R 3 for , or ;

[0019] X 1a For NR 3a O or S, and X 2a For N or CR 3a The premise is that if X 2a For CR 3a Then X 1a For NR 3a ;

[0020] X 1b and X 2b One of them is N, and the other is CR. 3b ;

[0021] X 2c For N or CR 3c ;

[0022] X 3a X4a X 5a and X 6a No more than one of them is N, and the rest are CR. 3d ;

[0023] X 3b X 4b X 5b and X 6b No more than one of them is N, and the rest are CR. 3e ;

[0024] X 3c X 4c X 5c and X 6c No more than one of them is NR 3f The rest are CR 3f R 3g ;

[0025] R 3a R 3b and R 3c Each is independently H or C1-C6 alkyl; and

[0026] R 3d R 3e R 3f and R 3g Each is independently H, halogen, cyano, hydroxyl, amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, C1-C6 alkoxy, or C1-C6 alkyl, wherein R 3d R 3e R 3f and R 3g Any C1-C6 alkoxy and C1-C6 alkyl groups may optionally be substituted with halogen, cyano, or hydroxyl groups.

[0027] Another object of the present invention is to provide a pharmaceutical composition comprising a novel compound that inhibits the activity of CDK12 and / or CDK13 and degrades cyclin K, for example, a pharmaceutical composition for treating cancer.

[0028] Another object of the present invention is to provide a method for inhibiting CDK12 and / or CDK13 and degrading cyclin K using novel compounds that inhibit the activity of CDK12 and / or CDK13 and degrade cyclin K; and the use of the method for inhibiting CDK12 and / or CDK13 and degrading cyclin K.

[0029] Another object of the present invention is to provide a method for treating diseases (e.g., cancer) associated with CDK12 and / or CDK13 and / or cyclin K using novel compounds that inhibit the activity of CDK12 and / or CDK13 and degrade cyclin K.

[0030] Technical Problem Solutions

[0031] Every description and embodiment disclosed in this application can also be applied to every other description and embodiment. That is, all combinations of the various elements disclosed in this application fall within the scope of this application. Furthermore, the scope of this application should not be considered limited to the specific embodiments described below.

[0032] In one aspect of the invention, compounds of formula I, stereoisomers thereof, hydrates, solvates or pharmaceutically acceptable salts are provided:

[0033] [Formula I]

[0034]

[0035] In Equation I of this paper, R 1 It is a halogenated group, hydroxyl group, cyano group, C1-C6 alkoxy group, C1-C6 alkyl group, C2-C6 alkenyl group, C2-C6 ynyl group, C3-C6 cycloalkyl group, C3-C6 cycloalkyl-C1-C6 alkyl-, C3-C6 cycloalkyl-C2-C6 alkenyl-, or C3-C6 cycloalkyl-C2-C6 ynyl-. In one embodiment, R 1 Or R 1 Any C1-C6 alkoxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, and C3-C6 cycloalkyl groups contained herein may be optionally substituted with halogen, hydroxyl, or cyano groups, each independently.

[0036] In one implementation, R 1 It can be a halogenated group, optionally substituted with a halogenated C1-C6 alkyl, C2-C6 alkenyl, C2-C6 ynyl, C3-C6 cycloalkyl, C3-C6 cycloalkyl-C1-C6 alkyl-, C3-C6 cycloalkyl-C2-C6 alkenyl-, or C3-C6 cycloalkyl-C2-C6 ynyl-. For example, R 1 It can be ethynyl, methylethynyl, cyclopropylethynyl, methyl, ethyl, isopropyl, cyclopropyl, trifluoromethyl, trifluoroethyl, iodo, or chloro.

[0037] In one implementation, R 1 It can be a C1-C6 haloalkyl group, such as CF3.

[0038] In Equation I of this paper, R 2 For C6-C10 Aryl; a 5- or 6-membered heteroaryl group containing 1 to 2 nitrogen atoms; or a 4- to 12-membered heterocyclic group containing 1 nitrogen atom and connected via the nitrogen atom to an imidazopyridazine ring of Formula I.

[0039] In one implementation, R in formula I 2 It can be a 4- to 12-membered heterocyclic group, which can be monocyclic, bridged, or spirocyclic. The 4- to 12-membered heterocyclic group may contain one nitrogen atom and may be linked via this nitrogen atom to an imidazopyridazine ring of Formula I. In one embodiment, the heterocyclic group may optionally contain one additional nitrogen atom or one oxygen atom.

[0040] In one embodiment, the 4- to 12-membered heterocyclic group may be a nitrogen-containing heterocyclic butyl, pyrrolidinyl, morpholinyl, piperidinyl, or piperazineyl. In one embodiment, any two non-adjacent carbon atoms of the morpholinyl, piperidinyl, or piperazineyl group may optionally be connected to each other via a C1-C3 alkylene group to form a bridged ring. For example, the bridged ring heterocyclic group can be 3-oxa-8-azabicyclo[3.2.1]octane-8-yl, 8-oxa-3-azabicyclo[3.2.1]octane-3-yl, 3,8-diazabicyclo[3.2.1]octane-3-yl, 3,8-diazabicyclo[3.2.1]octane-8-yl, 2-oxa-5-azabicyclo[2.2.2]octane-5-yl, 8-azabicyclo[3.2.1]octane-8-yl, 3-azabicyclo[3.1.1]heptane-3-yl, 6-oxa-3-azabicyclo[3.1.1]heptane-3-yl, 3,6-diazabicyclo[3.1.1]heptane-3-yl, or 3,6-diazabicyclo[3.1.1]heptane-6-yl, but is not limited thereto.

[0041] Alternatively, the 4- to 12-membered heterocyclic group can be a 7- to 11-membered azerosilicon ring containing a nitrogen atom attached to an imidazopyridazine ring of Formula I and optionally containing an additional nitrogen atom or an oxygen atom. For example, the 7- to 11-membered azerosilicon ring can be 2-azerosilicon[3.3]heptane-2-yl, 2,6-diazaspiro[3.3]heptane-2-yl, 6-oxa-2-azerosilicon[3.3]heptane-2-yl, 2-azerosilicon[3.4]octane-2-yl, 2,6-diazaspiro[3.4]octane-2-yl, 6-oxa-2-azerosilicon ... Octane-2-yl, 6-azaspiro[3.4]octane-6-yl, 2,6-diazaspiro[3.4]octane-6-yl, 2-oxa-6-azaspiro[3.4]octane-6-yl, 2-azaspiro[4.4]nonane-2-yl, 2,7-diazaspiro[4.4]nonane-2-yl, or 2-oxa-7-azaspiro[4.4]nonane-7-yl. For example, the azaspirocycle may include, but is not limited to, 2-azaspiro[3.3]heptane-2-yl, 2,6-diazaspiro[3.3]heptane-2-yl, and 6-oxa-2-azaspiro[3.3]heptane-2-yl.

[0042] Alternatively, R in Equation I of this paper 2 It can be a 5- or 6-membered heteroaryl group containing one or two nitrogen atoms. In one embodiment, the 5- or 6-membered heteroaryl group can be pyrimidinyl, pyridinyl, pyrroleyl, or imidazolyl. For example, the heteroaryl group can be pyrimidinyl.

[0043] Alternatively, R in Equation I of this paper 2 It can be C6-C 10 Aryl groups. For example, C6-C 10 The aryl group can be phenyl or naphthyl. For example, C6-C 10 The aryl group can be phenyl.

[0044] In equation I above, R 2 It may optionally be substituted by one or more substituents selected from the group consisting of:

[0045] (i) H, halogen, hydroxyl, cyano, C1-C6 alkyl, C3-C6 cycloalkyl,

[0046] (ii) C1-C6 alkyl groups substituted with halogen, hydroxyl, or cyano groups,

[0047] (iii) C3-C6 cycloalkyl groups substituted with halogen, hydroxyl, or cyano groups,

[0048] (iv) amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, (C3-C6 cycloalkyl)amino, (C1-C6 hydroxyalkyl)amino, (C1-C6 alkyl)carbonylamino, (C1-C6 hydroxyalkyl)carbonylamino, or (C1-C6 alkoxy)(C1-C6 alkyl)carbonylamino, and

[0049] (v) (C1-C6 alkyl)carbonyl, (C1-C6 hydroxyalkyl)carbonyl, aminocarbonyl, (C1-C6 alkyl)aminocarbonyl, di(C1-C6 alkyl)aminocarbonyl, carboxyl or (C1-C6 alkoxy)carbonyl.

[0050] In one implementation, R in formula I 2 It may not be substituted by the substituents listed in (i) to (v) above. Alternatively, R in formula I... 2 It can be substituted independently by one or two substituents selected from the substituents listed in (i) to (iv) above.

[0051] In one implementation, R in equation I above 2 It can be substituted with H, hydroxyl, cyano, C1-C6 alkyl, C3-C6 cycloalkyl, hydroxy-(C1-C6 alkyl), hydroxy-(C3-C6)cycloalkyl, amino, (C1-C6 alkyl)amino, (C3-C6 cycloalkyl)amino, (C1-C6 hydroxyalkyl)carbonylamino, (C1-C6 alkyl)carbonyl, (C1-C6 hydroxyalkyl)carbonyl, aminocarbonyl, (C1-C6 alkyl)aminocarbonyl, or carboxyl, but is not limited thereto. For example, R in Formula I above 2 It can be substituted by H, hydroxyl, cyano, methyl, ethyl, cyclopropyl, cyclobutyl, hydroxymethyl, hydroxyethyl, 2-hydroxyisopropyl, 1-hydroxycyclopropyl, 1-hydroxycyclobutyl, amino, methylamino, ethylamino, cyclopropylamino, hydroxyacetamido, methylcarbonyl (i.e., acetyl), hydroxyacetyl, aminocarbonyl, carbamoyl or carboxyl, but is not limited thereto.

[0052] In one implementation, R in Equation I of this document 2 For C6-C 10 aryl, wherein the aryl group may optionally be selected from the group consisting of R. 2d Substitution: H, hydroxyl, amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, and (C3-C6 cycloalkyl)amino. For example, R 2 It can be phenyl, and R 2d It can be hydroxyl or amino.

[0053] In one implementation, R in Equation I of this document 2It is a 5- or 6-membered heteroaryl group containing 1 to 2 nitrogen atoms, wherein the heteroaryl group may optionally be selected from the group consisting of R. 2d Substitution: H, hydroxyl, amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, and (C3-C6 cycloalkyl)amino. For example, R 2 It can be pyridinyl or pyrimidinyl, and R 2d It can be hydroxyl or amino.

[0054] In one implementation, R in Equation I of this document 2 It can be selected from either formula A or formula B below.

[0055] [Formula A]

[0056]

[0057] [Formula B]

[0058]

[0059] In formula A above, n1 and n2 are each independently 1 or 2. In one embodiment, the carbon atoms of the ring constituting formula A may optionally be substituted with C1-C6 alkyl groups.

[0060] In equation A above, if both n1 and n2 are 2, then Y 1 For CR 2a R 2b NR 2c Or O. In this case, any two non-adjacent carbon atoms in the ring of formula A may optionally be connected to each other via a C1-C3 alkylene group to form a bridged ring. Alternatively, if one or both of n1 and n2 are 1, then Y 1 For CR 2a R 2b .

[0061] In formula B above, n3, n4, n5, and n6 are each independently 1 or 2. For example, n3, n4, n5, and n6 can all be 1. In one embodiment, the carbon atoms of the ring constituting formula B may optionally be substituted with C1-C6 alkyl groups. In formula B above, Y 2 For CR 2a R 2b NR 2c Or O.

[0062] In equations A and B above, R 2a and R 2bEach can be independently selected from the group consisting of: H, halogen, hydroxyl, cyano, C1-C6 alkyl or C3-C6 cycloalkyl; C1-C6 alkyl substituted with halogen, hydroxyl or cyano; C3-C6 cycloalkyl substituted with halogen, hydroxyl or cyano; amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, (C3-C6 cycloalkyl)amino, (C1-C6 alkyl)carbonylamino, (C1-C6 hydroxyalkyl)carbonylamino; and (C1-C6 alkyl)carbonyl, (C1-C6 hydroxyalkyl)carbonyl, aminocarbonyl, (C1-C6 alkyl)aminocarbonyl, di(C1-C6 alkyl)aminocarbonyl and carboxyl.

[0063] In one implementation, in equations A and B above, R 2a and R 2b One of them can be selected from the group consisting of: H, halogen, hydroxyl, cyano, C1-C6 alkyl or C3-C6 cycloalkyl; C1-C6 alkyl substituted with halogen, hydroxyl or cyano; C3-C6 cycloalkyl substituted with halogen, hydroxyl or cyano; amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, (C3-C6 cycloalkyl)amino, (C1-C6 alkyl)carbonylamino, (C1-C6 hydroxyalkyl)carbonylamino; and (C1-C6 alkyl)carbonyl, (C1-C6 hydroxyalkyl)carbonyl, aminocarbonyl, (C1-C6 alkyl)aminocarbonyl, di(C1-C6 alkyl)aminocarbonyl and carboxyl. In this case, R 2a and R 2b The other group can be H, a halogen group, a hydroxyl group, a cyano group, or a C1-C6 alkyl group.

[0064] For example, in equations A and B above, R 2a and R 2b It can be H, hydroxyl, amino, cyclopropylamino, hydroxyacetamide, methyl, hydroxymethyl, 2-hydroxyisopropyl, hydroxycyclopropyl, carboxyl, carbamoyl, hydroxyacetyl, or cyano, but is not limited to these.

[0065] In equations A and B above, R 2c It can be H, C1-C6 alkyl, C3-C6 cycloalkyl, (C1-C6 alkyl)carbonyl, (C1-C6 hydroxyalkyl)carbonyl, aminocarbonyl, (C1-C6 alkyl)aminocarbonyl, or di(C1-C6 alkyl)aminocarbonyl. For example, R 2c It can be H, methyl, cyclopropyl, acetyl, hydroxyacetyl, or carbamoyl, but is not limited to these.

[0066] In one implementation, R in Equation I of this document 2 You can choose from the following groups:

[0067]

[0068]

[0069]

[0070] In R 2 In the structure, R 2a R 2b R 2c and R 2d This is the same as the description given above for equations A and B.

[0071] For example, R in Equation I of this paper 2 You can choose from the following groups:

[0072]

[0073]

[0074]

[0075]

[0076] .

[0077] In Equation I of this paper, R 3 Choose from the following groups:

[0078] (1) (2) Or (3) .

[0079] In (1), X 1a For NR 3a O or S, and X 2a For N or CR 3a The premise is that if X 2a For CR 3a Then X 1a For NR 3a In addition, X 3a X 4a X 5a and X 6a No more than one of them is N, and the rest are CR. 3d .

[0080] In (2), X 1b and X 2b One of them is N, and the other is CR. 3b In addition, X 3b X 4b X 5b and X 6bNo more than one of them is N, and the rest are CR. 3e .

[0081] In (3), X 2c For N or CR 3c In addition, X 3c X 4c X 5c and X 6c No more than one of them is NR 3f The rest are CR 3f R 3g .

[0082] In (1) to (3), R 3a R 3b and R 3c Each is independently H or C1-C6 alkyl. Furthermore, R 3d R 3e R 3f and R 3g Each is independently H, halogen, cyano, hydroxyl, amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, C1-C6 alkoxy, or C1-C6 alkyl. In this case, R 3d R 3e R 3f and R 3g Any C1-C6 alkoxy and C1-C6 alkyl groups may optionally be substituted with halogen, cyano, or hydroxyl groups.

[0083] In one implementation, R in Equation I of this document 3 You can choose from the following groups:

[0084]

[0085] In R 3 In the structure, X 3a X 4a X 5a and X 6a No more than one of them is N, and the rest are CR. 3d X 3b X 4b X 5b and X 6b No more than one of them is N, and the rest are CR. 3e X 3c X 4c X 5c and X 6c No more than one of them is NR 3f The rest are CR 3f R 3g .

[0086] In this case, R 3a R 3b and R 3c Each is independently H or C1-C6 alkyl. R 3d R 3e R 3f and R 3g Each can be independently H, halogen, cyano, hydroxyl, amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, C1-C6 alkoxy, or C1-C6 alkyl. 3d R 3e R 3f and R 3g Any C1-C6 alkoxy and C1-C6 alkyl groups may optionally be substituted with halogen, cyano, or hydroxyl groups. In one embodiment, R 3d R 3e R 3f and R 3g It can be H, a halogen group, a hydroxyl group, an amino group, a C1-C6 alkoxy group, a C1-C6 alkyl group, or a C1-C6 haloalkyl group. For example, R 3d R 3e R 3f and R 3g Each can be independently H, F, Cl, hydroxyl, amino, methoxy, methyl, and trifluoromethyl. For example, R 3d R 3e R 3f and R 3g They can be halogens, such as F or Cl, on their own.

[0087] In one implementation, R 3 You can choose from the following groups:

[0088] ; ; ; ; ; ; ; ; ; ; ; ; ; ;as well as .

[0089] In R 3 In the structure, R 3a R 3b and R 3c Each is independently an H or C1-C6 alkyl group.

[0090] In R3 In the structure, R 3d R 3e R 3f and R 3g Each can be independently H, a halogen group, a hydroxyl group, an amino group, a C1-C6 alkoxy group, a C1-C6 alkyl group, or a C1-C6 haloalkyl group. For example, R 3d R 3e R 3f and R 3g They can be H, F, Cl, hydroxyl, amino, methoxy, methyl, and trifluoromethyl, each independently.

[0091] For example, R in Equation I of this paper 3 You can choose from the following groups:

[0092]

[0093]

[0094] .

[0095] In one embodiment, the compound of formula I of the present invention may be a compound of formula IA:

[0096] [Form IA]

[0097]

[0098] In the above equation IA, R 1 It can be a halogenated group, hydroxyl group, cyano group, C1-C6 alkoxy group, C1-C6 alkyl group, C2-C6 alkenyl group, C2-C6 alkynyl group, C3-C6 cycloalkyl group, C3-C6 cycloalkyl-C1-C6 alkyl- group, C3-C6 cycloalkyl-C2-C6 alkenyl- group, or C3-C6 cycloalkyl-C2-C6 alkynyl- group. R 1 Or R 1 Any C1-C6 alkoxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, and C3-C6 cycloalkyl groups contained herein may optionally be substituted with a halogen, hydroxyl, or cyano group. The R described above with respect to Formula I... 1 Various examples can also be applied to formula IA.

[0099] In formula IA above, ring A is a 4- to 12-membered heterocyclic group that is monocyclic, bridged, or spirocyclic, and may optionally contain an additional nitrogen atom or an oxygen atom. The above refers to R in formula I. 2 The various examples of 4- to 12-membered heterocyclic groups of monocyclic, bridged, or spirocyclic forms described can also be applied to Formula IA.

[0100] In the above formula IA, ring A may optionally be substituted by one or more substituents selected from the group consisting of:

[0101] (i) H, halogen, hydroxyl, cyano, C1-C6 alkyl, C3-C6 cycloalkyl,

[0102] (ii) C1-C6 alkyl groups substituted with halogen, hydroxyl, or cyano groups,

[0103] (iii) C3-C6 cycloalkyl groups substituted with halogen, hydroxyl, or cyano groups,

[0104] (iv) amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, (C3-C6 cycloalkyl)amino, (C1-C6 hydroxyalkyl)amino, (C1-C6 alkyl)carbonylamino, (C1-C6 hydroxyalkyl)carbonylamino, or (C1-C6 alkoxy)(C1-C6 alkyl)carbonylamino, and

[0105] (v) (C1-C6 alkyl)carbonyl, (C1-C6 hydroxyalkyl)carbonyl, aminocarbonyl, (C1-C6 alkyl)aminocarbonyl, di(C1-C6 alkyl)aminocarbonyl, carboxyl or (C1-C6 alkoxy)carbonyl.

[0106] The above refers to R in equation I. 2 The various examples described by the substituents in Formula A and Formula B can also be applied to Formula IA.

[0107] In the above equation IA, R 3d It is H, halogen, cyano, hydroxyl, amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, C1-C6 alkoxy, or C1-C6 alkyl. In this case, R 3d Any C1-C6 alkoxy and C1-C6 alkyl groups may optionally be substituted with a halogen, cyano, or hydroxyl group. The above refers to R in Formula I. 3d The various examples described can also be applied to formula IA.

[0108] In the above formula IA, p can be 0, 1, 2, or 3. In one implementation, p can be 0, 1, or 2.

[0109] In one embodiment, the compound of formula I of the present invention can be a compound of formula IB:

[0110] [Form IB]

[0111]

[0112] In the above equation IB, R 1It can be a halogenated group, hydroxyl group, cyano group, C1-C6 alkoxy group, C1-C6 alkyl group, C2-C6 alkenyl group, C2-C6 alkynyl group, C3-C6 cycloalkyl group, C3-C6 cycloalkyl-C1-C6 alkyl- group, C3-C6 cycloalkyl-C2-C6 alkenyl- group, or C3-C6 cycloalkyl-C2-C6 alkynyl- group. R 1 Or R 1 Any C1-C6 alkoxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, and C3-C6 cycloalkyl groups contained herein may optionally be substituted with a halogen, hydroxyl, or cyano group. The R described above with respect to Formula I... 1 Various examples can also be applied to the IB formula.

[0113] In the above equation IB, ring B is selected from the following groups: In one implementation, ring B is .

[0114] In the above equation IB, R 2d It can be H, hydroxyl, amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, or (C3-C6 cycloalkyl)amino. For example, R 2d It may include hydroxyl or amino groups, but is not limited to these.

[0115] In the above equation IB, R 3d It is H, a halogen group, a cyano group, a hydroxyl group, a C1-C6 alkoxy group, or a C1-C6 alkyl group. In this case, R 3d Any C1-C6 alkoxy and C1-C6 alkyl groups may optionally be substituted with a halogen, cyano, or hydroxyl group. The above refers to R of formula I. 3d The various examples described can also be applied to Formula IB.

[0116] In the above formula IB, p can be 0, 1, 2, or 3. In one implementation, p can be 0, 1, or 2.

[0117] For example, in the above formula IB, It can be R 2d This is the same as described above for IB. For example, R 2d It can be amino, (C1-C6 alkyl)amino or di(C1-C6 alkyl)amino.

[0118] In one embodiment, the compound of formula I of the present invention may be a compound of formula IC-1 or formula IC-2.

[0119] [Formula IC-1]

[0120]

[0121] [Formula IC-2]

[0122]

[0123] In the above equation IC-1, n1 and n2 are each independently 1 or 2. (Containing Y) 1 The carbon atoms of the N ring can optionally be substituted with C1-C6 alkyl groups. In one embodiment, if both n1 and n2 are 2, then Y 1 For CR 2a R 2b NR 2c Or O. Alternatively, if one or both of n1 and n2 are 1, then Y 1 It is CR 2a R 2b .

[0124] In the above equation IC-2, Y 2 It is CR 2a R 2b NR 2c Or O.

[0125] In equations IC-1 and IC-2 above, R 1 It is a halogen group, hydroxyl group, cyano group, C1-C6 alkoxy group, C1-C6 alkyl group, or C1-C6 haloalkyl group.

[0126] In equations IC-1 and IC-2 above, R 3d Each can be independently H, halogen, hydroxyl, cyano, C1-C6 alkoxy, C1-C6 alkyl, or C1-C6 haloalkyl.

[0127] In equations IC-1 and IC-2 above, p is an integer from 0 to 2. For example, p is 2.

[0128] In equations IC-1 and IC-2 above, R 2a and R 2b Each is independently selected from the group consisting of: H, halogen, hydroxyl, cyano, C1-C6 alkyl or C3-C6 cycloalkyl; C1-C6 alkyl substituted with halogen, hydroxyl or cyano; C3-C6 cycloalkyl substituted with halogen, hydroxyl or cyano; amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, (C3-C6 cycloalkyl)amino, (C1-C6 alkyl)carbonylamino, (C1-C6 hydroxyalkyl)carbonylamino; and (C1-C6 alkyl)carbonyl, (C1-C6 hydroxyalkyl)carbonyl, aminocarbonyl, (C1-C6 alkyl)aminocarbonyl, di(C1-C6 alkyl)aminocarbonyl and carboxyl.

[0129] In equations IC-1 and IC-2 above, R 2cIt can be H, C1-C6 alkyl, C3-C6 cycloalkyl, (C1-C6 alkyl)carbonyl, (C1-C6 hydroxyalkyl)carbonyl, aminocarbonyl, (C1-C6 alkyl)aminocarbonyl or di(C1-C6 alkyl)aminocarbonyl.

[0130] Regarding n1, n2, and Y in formula IC-1 or IC-2 1 Y 2 R 1 R 2a R 2b R 2c R 3d And p, the specific examples described above for equations I, A, B, IA and IB can also be applied to the corresponding variables and structures of equations IC-1 or IC-2.

[0131] The compounds of Formula I can be compounds selected from the group consisting of the following.

[0132]

[0133]

[0134]

[0135]

[0136]

[0137]

[0138]

[0139]

[0140] Definitions

[0141] All technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art. Unless otherwise stated, conventional techniques, methods of measurement, manufacturing processes, and ingredients or materials are used, based on conventional techniques such as pharmacology, medicinal chemistry, mass spectrometry, NMR, HPLC, and biochemistry.

[0142] The features and components of each embodiment described and illustrated herein may be combined with features and components of any other embodiment without departing from the scope or spirit of this disclosure.

[0143] Unless otherwise specified, in this specification and the appended claims, "and" and "or" mean "and / or". The terms "comprising" and "including" are open-ended and mean that a compound, composition, or method may include additional features or components in addition to the specific features or components listed.

[0144] As used herein, the range of values ​​indicated by the term "to" refers to the range that includes the values ​​described before and after the term "to" as the lower and upper limits, respectively.

[0145] As used herein, the terms “optional” or “optionally” mean that an event or condition described below may or may not occur, and the description includes instances where the event or condition occurs as well as instances where the event or condition does not occur. For example, the term “optionally substituted” includes instances where the element is substituted by a specified substituent or is not substituted.

[0146] Compounds

[0147] The term "halogen" refers to an atom belonging to Group 17 of the periodic table. Halogen atoms include fluorine (F), chlorine (Cl), bromine (Br), and iodine (I). The term "halogen" is used interchangeably with the term "halogen group," which refers to a monovalent functional group composed of halogen atoms.

[0148] The term "hydroxyl group" refers to the -OH functional group (hydroxyl group).

[0149] The term "-CN" or "cyano" refers to a functional group consisting of a triple bond between a carbon atom and a nitrogen atom.

[0150] The term "amino" refers to the functional group in which hydrogen and nitrogen atoms are bonded, namely, -NH2.

[0151] As used herein, the term "alkylamino" refers to a functional group in which one hydrogen atom of an amino group is replaced by an alkyl group. For example, C 1-6 Alkylamino groups can include, but are not limited to, -NH(C) 1- C6 alkyl), such as methylamino, ethylamino, propylamino and butylamino.

[0152] As used herein, the term "dialkylamino" refers to a functional group in which two hydrogen atoms of an amino group are each substituted with an alkyl group. In this case, the substituted alkyl groups may be the same or different. For example, di(C 1-6 Alkyl)amino groups can include, but are not limited to, -N(C) 1- C6 alkyl)2, such as dimethylamino, diethylamino, dipropylamino, dibutylamino, ethylmethylamino, methylpropylamino and ethylpropylamino.

[0153] The term "oxo" refers to =O, and "oxo-substituted" means that the carbon atom has a =O substituent in the form of -C(=O)-. The carbon atom substituted by the oxo group can be provided as a carbonyl group.

[0154] As used in this article, the term "carbonyl" refers to a divalent functional group of -C(=O)-.

[0155] As used in this article, the term "carboxyl group" refers to -COOH.

[0156] The term "alkyl" refers to a fully saturated, branched or unbranched (or straight or linear) hydrocarbon group. Alkyl groups can be substituted or unsubstituted. Alkyl groups can be C1 to C8, C1 to C7, C1 to C6, C1 to C5, C1 to C4, C1 to C3, or C1 to C2 alkyl groups. Non-limiting examples of alkyl groups may include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, n-pentyl, isopentyl, neopentyl, isopentyl, or n-hexyl.

[0157] The term "haloalkyl" refers to a straight-chain or branched saturated aliphatic hydrocarbon group having a specified number of carbon atoms and being substituted with one or more halogen atoms. Haloalkyl includes perhaloalkyl, wherein all hydrogen atoms of the alkyl group are substituted with a halogen (e.g., -CF3, -CF2CF3). The aforementioned halogens may be the same (e.g., CHF2, -CF3) or different (e.g., CF2Cl). If specified, the haloalkyl may optionally be substituted with one or more substituents other than halogens. Examples of haloalkyl include, but are not limited to, fluoromethyl, dichloroethyl, trifluoromethyl, trichloromethyl, pentafluoroethyl, and pentachloroethyl.

[0158] As used herein, the term "alkylene" refers to a compound having the following structure: -C n H 2n - indicates a divalent, fully saturated, branched or unbranched (or straight or linear) hydrocarbon group. For example, C 1-6 Alkylenes can include ethylene, propyleneene, butylene, and hexylene.

[0159] The term "alkenyl" refers to a straight-chain or branched hydrocarbon group having 2 to 8 carbon atoms, and in some embodiments 2 to 6 carbon atoms or 2 to 4 carbon atoms, and having at least one vinyl unsaturated site (>C=C<). For example, (C x -C y Alkenyl refers to an alkenyl group having x to y carbon atoms, and may include, for example, vinyl, propenyl, isopropenyl, 1,3-butadienyl, etc.

[0160] The term "alkynyl" refers to a straight-chain or branched monovalent hydrocarbon group containing at least one triple bond. The term "alkynyl" can also include hydrocarbon groups having one triple bond and one double bond. For example, (C2-C6) alkynyl groups can include ethynyl, propynyl, etc.

[0161] The term "alkoxy" refers to a substituent in which a substituted or unsubstituted straight-chain or branched alkyl moiety is attached to another chemical structure via oxygen. Alkoxy groups can include, but are not limited to, all of their possible isomers, such as methoxy, ethoxy, propoxy, and butoxy, or isopropoxy, isobutoxy, and tert-butoxy.

[0162] The term "cycloalkyl" refers to a saturated or partially unsaturated hydrocarbon ring having a specified number of carbon atoms as the ring element. For example, C3-C6 cycloalkyl refers to a cycloalkyl ring having 3, 4, 5, or 6 carbon atoms as the ring element. As used herein, the term "cycloalkyl" can refer to, for example, C3-C6 cycloalkyl, C3-C5 cycloalkyl, or C3-C4 cycloalkyl. Examples may include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, or cyclohexenyl.

[0163] The term "heterocyclic group" or "heterocycle" refers to a saturated or partially unsaturated cyclic hydrocarbon group containing at least one heteroatom. Heterocyclic groups can be monocyclic, bicyclic, or tricyclic. The two cyclic groups described above can be spirocyclic, bridged, or fused rings. A spirocyclic group is a structure in which two rings share a common atom. A bridged ring group is a structure in which two non-adjacent ring elements are connected by one or more bridging elements. Heterocyclic groups can contain 3 to 20, 3 to 12, 3 to 10, 3 to 7, 3 to 6, 4 to 6, or 5 to 6 ring elements. Heterocyclic groups can contain one or more heteroatoms selected from the group consisting of N, O, and S.

[0164] One or more N or S atoms in a heterocyclic group can be oxidized, and the term "heterocyclic group" includes such oxidized forms (e.g., N → O-, S(O), SO2). Non-limiting examples of monocyclic heterocyclic groups may include aziridinyl, azircyclic butyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, and azircyclic heptyl. For example, spirocyclic heterocyclic groups may include 2-azaspiro[3.3]heptane-2-yl, 2,6-diazaspiro[3.3]heptane-2-yl, 6-oxa-2-azaspiro[3.3]heptane-2-yl, 2-azaspiro[3.4]octane-2-yl, 2,6-diazaspiro[3.4]octane-2-yl, 6-oxa-2-azaspiro[3.4]octane-2-yl. -yl, 6-azaspiro[3.4]octane-6-yl, 2,6-diazaspiro[3.4]octane-6-yl, 2-oxa-6-azaspiro[3.4]octane-6-yl, 2-azaspiro[4.4]nonane-2-yl, 2,7-diazaspiro[4.4]nonane-2-yl or 2-oxa-7-azaspiro[4.4]nonane-7-yl, but not limited thereto. For example, bridged ring heterocyclic groups may include, but are not limited to, 3-oxa-8-azabicyclo[3.2.1]octane-8-yl, 8-oxa-3-azabicyclo[3.2.1]octane-3-yl, 3,8-diazabicyclo[3.2.1]octane-3-yl, 3,8-diazabicyclo[3.2.1]octane-8-yl, 2-oxa-5-azabicyclo[2.2.2]octane-5-yl, 8-azabicyclo[3.2.1]octane-8-yl, 3-azabicyclo[3.1.1]heptane-3-yl, or 6-oxa-3-azabicyclo[3.1.1]heptane-3-yl, 3,6-diazabicyclo[3.1.1]heptane-3-yl, or 3,6-diazabicyclo[3.1.1]heptane-6-yl.

[0165] The term "aryl" also includes aromatic rings, or groups formed by the fusion of an aromatic ring with one or more carbon rings. (C6-C) 12 The aryl group can be, for example, C6 to C6. 10 Aryl or C6 to C8 aryl. Non-limiting examples of aryl groups may include phenyl or naphthyl.

[0166] The term "heteroaryl" refers to a monocyclic or bicyclic group containing one or more heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur, with the remaining ring atoms being carbon. A heteroaryl group may contain, for example, one to three heteroatoms and may contain five to ten ring elements. For example, a heteroaryl group may be a 5- or 6-membered monocyclic heteroaryl group. A heteroaryl group may contain, for example, one or two nitrogen atoms. S or N may be oxidized to have a variety of oxidation states. Examples of monocyclic heteroaryl groups may include, but are not limited to, pyrimidinyl, pyridine, pyrroleyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc.

[0167] In the phrase “optionally substituted” above, the term “substitution” means the introduction of a substituent for a hydrogen atom in the formation of a derivative by substituting one or more hydrogen atoms with another atomic group in an organic compound, and “substituent” refers to the introduced atomic group. As used herein, the term “substituted” means a group in which one or more hydrogen atoms are replaced by one or more non-hydrogen atomic groups, provided that the valence requirement is met and that a chemically stable compound is produced from the substitution. In this specification, unless explicitly stated as “unsubstituted,” all substituents should be interpreted as capable of being unsubstituted or substituted.

[0168] In this specification, when a combination of substituents (such as haloalkyl, hydroxyalkyl, etc.) is referred to as a group, the last group mentioned usually contains atoms attached to the remainder of the molecule.

[0169] As used in this article, " The symbols “”, “*”, or “-” are used to indicate the position of a substituent bonded to a portion of the compound. For example, if a “-” is indicated at the end of a substituent, it means that the end is attached to the remainder of the compound. Furthermore, when two or more substituents are connected by a “-”, it means that the substituent immediately preceding the “-” is bonded to the substituted atom of the substituent immediately following the “-”.

[0170] As used herein, the term "solvent" can refer to a compound of the present invention or a salt thereof comprising a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces. Therefore, preferred solvents may be volatile, non-toxic, and / or suitable for human administration. Those skilled in the art will be able to readily prepare solvates (e.g., hydrates) of the compounds disclosed herein using appropriate techniques known in the art.

[0171] As used herein, the term “stereoisomer” can refer to a compound of the present invention or a salt thereof having the same chemical formula or molecular formula but different in optical or stereostructure, and specifically can be a diastereomer, enantiomer or geometric isomer.

[0172] In some embodiments, the compounds of the present invention contain one or more asymmetric centers and may be in the form of racemic mixtures, single enantiomers, mixtures of enantiomers, single diastereomers, mixtures of diastereomers, etc. In one embodiment, due to the nature of the asymmetric centers or restricted rotation, the compounds of the present invention may exist in the form of enantiomers or diastereomers.

[0173] When two or more asymmetric centers are present in the compounds of this invention, multiple diastereomers and enantiomers with the chemical structures disclosed herein may exist. Pure isomers, isolated isomers, partially pure isomers, or racemic mixtures are all intended to fall within the scope of this invention.

[0174] Purification of isomers and separation of mixtures of isomers can be achieved using standard techniques known in the art. For example, mixtures of diastereomers can be separated into their respective diastereomers by chromatography or crystallization, and racemic mixtures can be separated into their respective enantiomers by chromatography or resolution of a chiral phase.

[0175] The term "salt" refers to both inorganic and organic acid addition salts of compounds. The compounds of this invention can be used in the form of pharmaceutically acceptable salts derived from inorganic or organic acids. Pharmaceutically acceptable salts are those that do not cause significant irritation to the organism administering the compound and do not impair the compound's biological activity and physical properties. Inorganic acid salts can be hydrochlorides, bromates, phosphates, sulfates, or hydrogen sulfates. Organic acid salts can be formates, acetates, propionates, lactates, oxalates, tartrates, malates, maleates, citrates, fumarates, benzenesulfonates, camphorsulfonates, ethanedisulfonates, trichloroacetates, trifluoroacetates, benzoates, gluconates, methanesulfonates, glycolates, succinates, 4-toluenesulfonates, galacturonic acids, dihydroxynaphthylates, glutamates, ethanesulfonates, benzenesulfonates, p-toluenesulfonates, or aspartate salts. Furthermore, metal salts can be calcium, sodium, magnesium, strontium, or potassium salts.

[0176] Pharmaceutically acceptable salts of the compounds according to the invention can be prepared by dissolving a compound of formula I in a water-miscible organic solvent (e.g., acetone, methanol, ethanol, acetonitrile, etc.), adding an excess of an organic acid or an aqueous acid solution containing an inorganic acid, and then precipitating or crystallizing. Subsequently, after evaporating the solvent or excess acid from the mixture, the pharmaceutically acceptable salt can be prepared by drying to obtain an addition salt or by filtration of the precipitated salt.

[0177] The compounds of the present invention (including stereoisomers, hydrates, solvates and salts) can be prepared by known organic synthesis methods and can be synthesized via a variety of synthetic routes.

[0178] The reactions used to prepare the compounds of this invention can be carried out in suitable solvents that can be appropriately selected by those skilled in the art of organic synthesis. A suitable solvent is one that does not substantially react with the starting materials (reactants), intermediates, or target products at the temperature at which the reaction occurs. Those skilled in the art will be able to appropriately select suitable solvents for each reaction step.

[0179] During the synthesis of the compounds of this invention, various functional groups may be protected and deprotected. Those skilled in the art will be able to readily determine the necessity of protection and deprotection and select appropriate protecting groups.

[0180] Each reaction can be monitored by any suitable method known in the art. For example, it can be monitored by spectroscopic means (e.g., NMR). 1 H or 13 C) Mass spectrometry or chromatography (HPLC or TLC) can be used to monitor the synthesis of the desired compound.

[0181] The compounds of the present invention can be synthesized according to the synthesis procedures described in the examples below. Based on the above, depending on the desired structure of the compound, the desired compound can be prepared by appropriately changing the reactants and reaction conditions, etc.

[0182] Medical uses, pharmaceutical compositions, and methods of administration

[0183] In another aspect, a pharmaceutical composition is provided comprising a compound, its stereoisomer, hydrate, solvate, or pharmaceutically acceptable salt according to one aspect. The compound, stereoisomer, solvate, and salt are the same as described above.

[0184] The compounds according to one aspect of the present invention can inhibit the overexpression, excessive activity, mutation of CDK12 and / or CDK13, as well as the activation of signaling pathways associated with cyclin K. Therefore, the compounds can inhibit cell proliferation, suppress cell cycle progression, and / or promote cell death.

[0185] CDK12 and its ortholog CDK13 belong to the CDK family, which regulates transcription and post-transcriptional processes. CDK12 and CDK13 are known to mediate gene transcription by forming a complex with cyclin K and phosphorylating the C-terminal domain of RNA polymerase II (e.g., Greifenberg et al., 2016, "Structural and Functional Analysis of the CDK13 / Cyclin K Complex," Cell Rep. Vol. 14, No. 2, pp. 320-331). The phosphorylation of Ser2 in the C-terminal domain of RNA polymerase II by the CDK12 / cyclin K and CDK13 / cyclin K complexes is considered a key step in the transition from transcription initiation to elongation.

[0186] Recently, several compounds have been reported to directly degrade cyclin K by linking CDK12-cyclin K to the DDB1-CUL4-RBX1E3 ligase (Zuzanna et al., 2023, "Design principles for cyclin K molecular glue degraders", Nat Chem Biol Sep 7). Therefore, the cyclin K degradation capabilities of these compounds are expected to exhibit characteristics quite different from those of conventional kinase inhibitors that regulate the phosphorylation of downstream signaling molecules.

[0187] The compounds according to the present invention can exhibit cyclin K degradation activity and CDK12 and / or CDK13 inhibitory activity.

[0188] In addition to directly inhibiting CDK12 and / or CDK13 in cells, cyclin K degradation can induce novel anticancer effects. Cyclin K is an essential partner for both CDK12 and / or CDK13 and is necessary for the activity of both. Independent of CDK12 and / or CDK13, cyclin K expression is increased in various cancer cells compared to normal cells, suggesting that cyclin K itself is directly involved in the proliferation of various cancer cells. Furthermore, cyclin K has been shown to increase the expression of specific proteins associated with treatment resistance, which may enhance the response to existing therapeutic agents (Yi Xiao and Jixin Dong, 2023, "Coming of Age: Targeting CyclinK in Cancers", Cells. Aug 11;12(16):2044).

[0189] Therefore, since the compounds according to the invention exhibit enhanced and sustained efficacy compared to CDK12 / CDK13 inhibitors that only inhibit kinase activity, the compounds according to the invention are expected to exhibit superior anticancer efficacy compared to existing therapeutic agents.

[0190] Therefore, the pharmaceutical composition according to one aspect of the present invention can be used to treat diseases caused by overexpression, excessive activity, mutation of CDK12 and / or CDK13 and / or activation of signaling pathways associated with cyclin K. Furthermore, the pharmaceutical composition can be used to inhibit cell proliferation, inhibit cell cycle progression, and / or promote cell death.

[0191] In one implementation, the disease can be cancer, a proliferative disease, a neurodegenerative disease, an autoimmune disease, or a disease caused by abnormal intracellular protein translation function.

[0192] As used herein, the term “proliferative disorder” refers to a disease characterized by excessive cell proliferation. Proliferative disorders are associated with: (1) pathological proliferation of normal quiescent or proliferating cells; (2) pathological migration of cells from their normal location (e.g., metastasis of tumor cells); (3) pathological expression of proteolytic enzymes (e.g., matrix metalloproteinases (e.g., collagenase, gelatinase, and elastase)) that may lead to undesirable transformation of the cellular matrix; and / or (4) pathological angiogenesis that occurs in proliferative retinopathy and tumor metastasis. Exemplary proliferative disorders include cancer, benign tumors, and angiogenesis that accompanies and promotes the disease state (defined above as pathological angiogenesis).

[0193] In one embodiment, the pharmaceutical composition may be a pharmaceutical composition for treating cancer. In one embodiment, the cancer may be cancer caused by overexpression, overactivity, mutation, and / or activation of signaling pathways associated with cyclin K of CDK12 and / or CDK13. For example, cancer may be cancer caused by overexpression, overactivity, mutation, and / or activation of signaling pathways associated with cyclin K of CDK12 and / or CDK13, such as cancer characterized by amplification of the CDK12, CDK13, or cyclin K genes.

[0194] In one implementation, cancer can be selected from the group consisting of:

[0195] (1) Cancers of the breast, liver, lung, colon, kidney and bladder, including small cell lung cancer, non-small cell lung cancer, head and neck cancer, thyroid cancer, esophageal cancer, gastric cancer, pancreatic cancer, ovarian cancer, gallbladder cancer, cervical cancer, prostate cancer and skin cancer (including squamous cell carcinoma).

[0196] (2) Sarcomas, such as osteosarcoma and osteoblastoma (bone), chondrosarcoma (cartilage), leiomyosarcoma (smooth muscle), rhabdomyosarcoma (skeletal muscle), mesothelioma and mesothelioma (serosarcoid lining of body cavities), fibrosarcoma (fibrous tissue), angiosarcoma or hemangioendothelioma (blood vessels), liposarcoma (adipose tissue), glioma and astrocytoma (neurogenic connective tissue present in the brain), myxosarcoma (primordial embryonic connective tissue) or mesenchymal and mixed mesodermal tumors (mixed connective tissue types);

[0197] (3) Hematopoietic tumors of the lymphatic system, such as leukemia, acute lymphoblastic leukemia, Hodgkin lymphoma, non-Hodgkin lymphoma, B-cell lymphoma, T-cell lymphoma, piloblastic lymphoma, myeloma, mantle cell lymphoma and Burkitt lymphoma;

[0198] (4) Myeloid hematopoietic tumors, such as acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), myelodysplastic syndrome (MDS) and promyelocytic leukemia;

[0199] (5) Tumors of the central and peripheral nervous systems, such as astrocytoma, neuroblastoma, glioma and schwannoma.

[0200] In one implementation, the neurodegenerative disease can be Alzheimer's disease, Parkinson's disease, or Lou Gehrig's disease. For example, an autoimmune disease can be rheumatoid arthritis, systemic lupus erythematosus, psoriasis, or Sjögren's syndrome. For example, a disease caused by abnormal intracellular protein translation can be muscle atrophy, myotonic dystrophy, amyotrophic lateral sclerosis, spinal muscular atrophy, or Fragile X syndrome.

[0201] As used herein, the term “treating” means suppressing a disease, such as suppressing the pathology or signs of a disease, symptom, or disorder of a person experiencing or exhibiting such a disease, symptom, or disorder, i.e., preventing the further development of the pathology and / or signs, or improving a disease, such as improving the pathology or signs of a disease, symptom, or disorder of a person experiencing or exhibiting such a disease, symptom, or disorder, i.e., reversing the pathology and / or signs, such as reducing the severity of the disease.

[0202] "Cancer" is a disease that can be prevented or treated by pharmaceutical compositions. It refers to a general term for diseases caused by cells exhibiting invasive characteristics (where cells divide and grow unchecked despite normal growth constraints), infiltrative characteristics (where cells infiltrate surrounding tissues), and metastatic characteristics (where cells spread to other parts of the body). The cancers for which the compounds of this invention have therapeutic activity can include any cancer that can be treated, alleviated, delayed, inhibited, or prevented, such as cancers caused by overexpression, excessive activity, mutation, and / or activation of signaling pathways associated with cyclin K, such as CDK12 and / or CDK13.

[0203] For example, cancer may include, but is not limited to, breast cancer, ovarian cancer, colorectal cancer, lung cancer, prostate cancer, stomach cancer, liver cancer, kidney cancer, bladder cancer, pancreatic cancer, head and neck cancer, thyroid cancer, skin cancer, bile duct cancer, esophageal cancer, hematopoietic tumors of the lymphatic and myeloid systems, tumors of the central and peripheral nervous systems, and sarcomas. In one embodiment, cancer may include breast cancer, lung cancer, stomach cancer, pancreatic cancer, or colorectal cancer. In one embodiment, cancer may be breast cancer, such as triple-negative breast cancer.

[0204] For example, the pharmaceutical compositions described herein can be used to treat “high-grade” cancers (e.g., high-grade serous ovarian cancer, metastatic non-small cell lung cancer, metastatic breast cancer (e.g., triple-negative breast cancer), metastatic gastric cancer, metastatic liver cancer), tumors exhibiting a specific phenotype (e.g., estrogen receptor-positive (ER+) breast cancer, human epidermal growth factor receptor 2 (HER2)-positive breast cancer or gastric cancer, tyrosine kinase-positive non-small cell lung cancer or PD-L1-positive solid tumors), and / or cancers that have developed resistance to previously administered therapeutic agents (e.g., chemotherapy agents (e.g., cisplatin or fluorouracil, CDK4 / 6 inhibitors (e.g., palbociclib), estrogen receptor degraders (e.g., fulvestrant), or PARP inhibitors (e.g., olaparib)).

[0205] The pharmaceutical composition may contain additional anticancer agents. The pharmaceutical composition may be a single composition or a combination of individual components. For example, according to one aspect, the pharmaceutical composition may be an oral dosage form, and the anticancer agent may be a parenteral dosage form.

[0206] When used for cancer treatment, the compounds of the present invention can be used alone or in combination with conventional surgery, radiotherapy, chemotherapy or immunotherapy.

[0207] For example, the compounds of this invention can be administered in combination with other anticancer therapies, such as radiotherapy, taxane derivatives (e.g., paclitaxel, docetaxel, cabazitaxel), platinum compounds (e.g., cisplatin, carboplatin), antimetabolites (e.g., 5-FU, gemcitabine, cytarabine), anti-CTLA4 therapy (e.g., ipilimumab, trimemumab), anti-PD1 therapy (e.g., nivolumab, pembrolizumab), anti-PD-L1 therapy (e.g., atezolizumab, durvalumab), anti-VEGF therapy (e.g., bevacizumab, ramucirumab, aflibercept), and anti-EGFR therapy (e.g., ... Cetuximab), topoisomerase inhibitors (e.g., irinotecan), anti-HER2 therapies (e.g., trastuzumab, pertuzumab), anti-hormonal therapies (e.g., tamoxifen, exemestane, letrozole, anastrozole), estrogen receptor inhibitors (e.g., ellastrant, fulvestrant), ERK inhibitors (e.g., ulitinib), PARP inhibitors (e.g., olaparib, niraparib, taprazolepanib), mTOR inhibitors (e.g., everolimus, tesimolimus), CDK4 / 6 inhibitors (e.g., abecilide, palbociclib), EGFR inhibitors (e.g., afatinib, ... Erlotinib, osimertinib, gefitinib, dacomitinib), HER2 inhibitors (e.g., neratinib, lapatinib), ALK inhibitors (e.g., crizotinib, alectinib, brigatinib, ceritinib), tyrosine kinase inhibitors (e.g., avatinib, ripretinib, sunitinib, sorafenib, pazopanib, regorafenib, cabozantinib, lenvatinib), MEK inhibitors (e.g., trametinib), BCR-ABL inhibitors (e.g., imatinib, nilotinib, dasatinib), PI3K inhibitors (e.g., apeliximab), FGFR inhibitors (e.g., futribatib, pembrolizumab). Gatitinib), ROS1 inhibitors (e.g., crizotinib, entrectinib), androgen biosynthesis inhibitors (e.g., abiraterone acetate), androgen receptor inhibitors (e.g., enzalutamide, darotamide, apatamide), Hedgehog inhibitors (e.g., sonidazole, vemodazole), MET inhibitors (e.g., carmatinib, terpoxtinib), AXL inhibitors, NTRK1 inhibitors, RET inhibitors (e.g., plascitinib, serpatinib), KRAS inhibitors (e.g., adagraxibub, sotorasibub), or RAF inhibitors (e.g., encofenib, vemurafenib).

[0208] Pharmaceutical compositions may contain pharmaceutically acceptable carriers. The term "carrier" is used to mean that it contains excipients, diluents, or adjuvants. For example, a carrier may be selected from the group consisting of: lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, polyvinylpyrrolidone, water, physiological saline, buffers such as PBS, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil. The composition may contain fillers, anticoagulants, lubricants, wetting agents, flavoring agents, emulsifiers, preservatives, or combinations thereof.

[0209] Pharmaceutical compositions can be prepared into any dosage form using conventional methods. For example, the composition can be formulated into an oral dosage form (e.g., powder, tablet, capsule, syrup, pill, or granule) or a parenteral dosage form (e.g., injection). In addition, the composition can be formulated into a systemic dosage form or a topical dosage form.

[0210] In pharmaceutical compositions, solid dosage forms for oral administration can be tablets, pills, powders, granules, or capsules. Solid dosage forms may also contain excipients. Examples of such excipients may include starch, calcium carbonate, sucrose, lactose, or gelatin. Additionally, solid dosage forms may contain lubricants such as magnesium stearate or talc. In pharmaceutical compositions, liquid dosage forms for oral administration can be suspensions, oral solutions, emulsions, or syrups. Liquid dosage forms may contain water or liquid paraffin. Liquid dosage forms may contain excipients such as wetting agents, sweeteners, flavoring agents, or preservatives. In pharmaceutical compositions, dosage forms for parenteral administration can be sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, or suppositories. Non-aqueous solutions or suspensions may contain vegetable oils or esters. Vegetable oils may be, for example, propylene glycol, polyethylene glycol, or olive oil. Esters may be, for example, ethyl oleate. The suppository base can be Wipedsol, polyethylene glycol, Tween 61, cocoa butter, glyceryl laurate oil, or glycerin gelatin.

[0211] A pharmaceutical composition comprises a compound, its stereoisomer, solvate, or a pharmaceutically acceptable salt thereof, as the active ingredient of the pharmaceutical composition. The term "active ingredient" refers to a physiologically active substance used to achieve pharmacological activity (e.g., cancer).

[0212] Pharmaceutical compositions may comprise an effective amount of a compound, its stereoisomers, solvates, or pharmaceutically acceptable salts according to one aspect. The term "effective amount" refers to an amount sufficient to exert a preventive or therapeutic effect when administered to a subject requiring prevention or treatment. The effective amount can be suitably selected by those skilled in the art based on the selected cells or subject. Preferred doses of the pharmaceutical composition vary depending on the subject's condition and weight, the severity of the disease, the form of the drug, the route of administration, and the duration of administration, but can be suitably selected by those skilled in the art. However, the compound, its stereoisomers, solvates, or pharmaceutically acceptable salts may be administered in divided doses, for example, from about 0.0001 mg / kg to about 100 mg / kg, or from about 0.001 mg / kg to about 100 mg / kg, 1 to 24 times daily, 1 to 7 times every 2 days to 1 week, or 1 to 24 times every 1 to 12 months. In a pharmaceutical composition, the content of the compound, its stereoisomers, solvates, or pharmaceutically acceptable salts may be from about 0.0001 wt% to about 10 wt% based on the total weight of the entire composition, or from about 0.001 wt% to about 1 wt%.

[0213] The method of administration can be oral or parenteral. Examples of routes of administration include oral, transdermal, subcutaneous, rectal, intravenous, intra-arterial, intraperitoneal, intramuscular, intrasternal, local, intranasal, intratracheal, or intradermal routes. The composition can be administered systemically or locally, and can be administered alone or in combination with other pharmaceutically active compounds.

[0214] In another aspect, a method is provided for treating a disease caused by overexpression, hyperactivity, mutation, and / or activation of associated signaling pathways of CDK12 and CDK13, the method comprising administering to a subject a compound, its stereoisomer, hydrate, solvate, or pharmaceutically acceptable salt according to one aspect. The compound, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt, and the treatable disease are the same as described above.

[0215] For example, the pharmaceutical compositions described herein can be used to treat “high-grade” cancers (e.g., high-grade serous ovarian cancer, metastatic non-small cell lung cancer, metastatic breast cancer (e.g., triple-negative breast cancer), metastatic gastric cancer, metastatic liver cancer), tumors exhibiting a specific phenotype (e.g., estrogen receptor-positive (ER+) breast cancer, human epidermal growth factor receptor 2 (HER2)-positive breast cancer or gastric cancer, tyrosine kinase-positive non-small cell lung cancer or PD-L1-positive solid tumors), and / or cancers that have developed resistance to previously administered therapeutic agents (e.g., chemotherapy agents (e.g., cisplatin or fluorouracil, CDK4 / 6 inhibitors (e.g., palbociclib), estrogen receptor degraders (e.g., fulvestrant), or PARP inhibitors (e.g., olaparib)).

[0216] In one embodiment, the method may include the steps of: determining whether the subject has high-grade cancer or tumor cells with a specific phenotype, or whether resistance has been developed to a previously applied therapeutic agent; and if it is determined that the subject has high-grade cancer or tumor cells with a specific phenotype, or has developed resistance, then administering the compound of the present invention.

[0217] The subject can be a mammal, such as a human, mouse, rat, cow, horse, pig, dog, monkey, sheep, goat, ape, or cat. The subject can be someone who has cancer-related symptoms or is at high risk of having cancer-related symptoms.

[0218] The method may further include administering to a subject a known active ingredient for treating cancer. The known active ingredient may be administered to the subject simultaneously, separately, or sequentially, along with a compound, its stereoisomer, hydrate, solvate, or pharmaceutically acceptable salt according to one aspect.

[0219] The administration method can be oral or parenteral. Examples of routes of administration include oral, transdermal, subcutaneous, rectal, intravenous, intra-arterial, intraperitoneal, intramuscular, intrasternal, local, intranasal, intratracheal, or intradermal routes.

[0220] In another aspect, a method for inhibiting CDK12 and / or CDK13 and degrading cyclin K is provided, the method comprising adding a compound, its stereoisomer, hydrate, solvate, or pharmaceutically acceptable salt according to one aspect to cells. In one embodiment, the method may be performed in vitro. The compound, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt, and treatable disease are the same as described above.

[0221] Invention Effects

[0222] This invention provides, according to one aspect, the use of compounds thereon for inhibiting the activity of CDK12 and / or CDK13, pharmaceutical compositions comprising the compounds, and their use for treating diseases (e.g., cancer) associated with CDK12 and / or CDK13. According to the invention, these compounds exhibit excellent inhibitory activity and selectivity against CDK12 and / or CDK13, and are therefore suitable for treating cancers such as breast cancer, gastric cancer, lung cancer, pancreatic cancer, and colorectal cancer. Attached Figure Description

[0223] Figure 1 This is a graph showing the results of measuring the cyclin K degradation ability of the compound of Example 8 in HCC70 breast cancer cells.

[0224] Figure 2 This is a graph showing the results of measuring the cyclin K degradation capacity of the compound of Example 30 in HCC70 breast cancer cells.

[0225] Figure 3 This is a graph showing the results of measuring the cyclin K degradation capacity of the compound of Example 40 in HCC70 breast cancer cells.

[0226] Figure 4 This is a graph showing the results of measuring the cyclin K degradation ability of the compound of Example 8 in MKN45 gastric cancer cells.

[0227] Figure 5 This is a graph showing the results of measuring the cyclin K degradation ability of the compound of Example 40 in MKN45 gastric cancer cells.

[0228] Figure 6 This is a graph showing how the compound of Example 30 resulted in a reduction in relative tumor size in an HCC70 zebrafish xenograft breast cancer model.

[0229] Figure 7 and Figure 8 The figure shows that the compound of Example 8 resulted in a reduction in relative tumor size and a decrease in the number of metastatic cancer cells in an AGS zebrafish xenograft gastric cancer model. Detailed Implementation

[0230] The present invention will be described in more detail through the following embodiments. However, these embodiments are for illustrative purposes, and the scope of the invention is not limited to these embodiments.

[0231] The abbreviations used in the following embodiments have the following meanings, and abbreviations not listed below have the meanings commonly used in the relevant fields.

[0232] NCS: N-chlorosuccinimide

[0233] NBS: N-bromosuccinimide

[0234] NIS: N-iodosuccinimide

[0235] MeCN: Acetonitrile

[0236] THF: Tetrahydrofuran

[0237] BnBr: Benzyl bromide

[0238] PMB: p-methoxybenzyl

[0239] Cbz: benzyloxycarbonyl

[0240] NaBH(OAc)3: Sodium triacetoxyborohydride

[0241] CbzOSu: N-(benzyloxycarbonyloxy)succinimide

[0242] TBDMSCl: tert-butyldimethylchlorosilane

[0243] TBDPSCl: tert-butyldiphenylchlorosilane

[0244] DMAP: 4-Dimethylaminopyridine

[0245] TBAF: Tetra-N-Butylammonium Fluoride

[0246] HATU: 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate

[0247] HOBT: 1-Hydroxybenzotriazole

[0248] EDCI: 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide

[0249] DIEA: N,N-Diisopropylethylamine

[0250] DMF: N,N-dimethylformamide

[0251] DCM: Dichloromethane

[0252] DMSO: Dimethyl sulfoxide

[0253] TFA: Trifluoroacetic acid

[0254] AcOH: Acetic acid

[0255] EtOAc: Ethyl acetate

[0256] NMP: N-methyl-2-pyrrolidone

[0257] HMPA: Hexamethylphosphoric triamine

[0258] CDI: 1,1'-carbonyldiimidazole

[0259] SEMCl: 2-(trimethylsilyl)ethoxymethyl chloride

[0260] TsOH: p-Toluenesulfonic acid

[0261] Na2SO4: Sodium sulfate

[0262] CuI: Cuprous iodide (I)

[0263] Pd / C: Palladium on carbon

[0264] Pd(OH)2 / C: Palladium hydroxide on carbon

[0265] RuPhos-Pd-G3: (2-Dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II)methanesulfonate

[0266] NaH: Sodium hydride

[0267] TEA: Triethylamine

[0268] Pd(PPh3)4: Tetra(triphenylphosphine)palladium(0)

[0269] Ti(OEt)4: Titanium ethoxide (IV)

[0270] Ti(i-PrO)4: Titanium isopropoxide (IV)

[0271] Pd(PPh3)2Cl2: Dichlorobis(triphenylphosphine)palladium(II)

[0272] NaBH4: Sodium borohydride

[0273] NaOH: Sodium hydroxide

[0274] Pd(dppf)Cl2: [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride

[0275] Preparation of intermediate S1: N-benzyl-N-(6-morpholinylimidazo[1,2-b]pyridazine-8-yl)glycine tert-butyl ester

[0276]

[0277] Step 1: Preparation of tert-butyl (6-chloroimidazo[l,2-b]pyridazin-8-yl)glycinate

[0278] 8-Bromo-6-chloro-imidazo[1,2-b]pyridazine (24 g, 103.24 mmol) was dissolved in glycine tert-butyl ester (240 mL) and stirred at 120 °C for 16 hours. Water (2000 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (800 mL). The organic layer was washed three times with brine (2000 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give (6-chloroimidazo[1,2-b]pyridazine). b 1,8-pyridazine-8-yl)glycine tert-butyl ester (26.6 g, yield 91.13%). MS: m / z = 283.1 (M+1, ESI+).

[0279] Step 2: Preparation of tert-butyl N-benzyl-N-(6-chloroimidazo[l,2-b]pyridazin-8- yl)glycinate

[0280] The product obtained in step 1 (26.3 g, 93.02 mmol) and benzyl bromide (25.2 mL, 212 mmol) were dissolved in MeCN (200 mL), followed by the addition of cesium carbonate (92.2 g, 283 mmol) and stirring at 60 °C for 16 hours. Water (1200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (300 mL). The organic layer was washed three times with brine (1200 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-benzyl-N-(6-chloroimidazolo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (24.22 g, yield 68.25%). MS: m / z = 373.1 (M+1, ESI+).

[0281] Step 3: Preparation of tert-butyl N-benzyl-N-(6-morpholinoimidazo[l,2-b]pyridazin-8- yl)glycinate

[0282] The product obtained in step 2 (14.2 g, 38.09 mmol) and morpholine (33.3 mL, 380.9 mmol) were dissolved in 1,4-dioxane (300 mL), followed by the addition of cesium carbonate (37.23 g, 114.3 mmol) and RuPhos-Pd-G3 (3.19 g, 3.81 mmol) and stirring at 110 °C for 16 hours. Water (900 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (300 mL). The organic layer was washed three times with brine (150 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give intermediate S1 (11.8 g, 73.16% yield). MS: m / z = 424.2 (M+1, ESI+).

[0283] Preparation of intermediate S2: N-benzyl-N-(3-bromo-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester

[0284]

[0285] Intermediate S1 (5.31 g, 12.54 mmol) was dissolved in THF (80 mL), and NBS (2.23 g, 12.54 mmol) was added in portions at -60 °C, and the mixture was stirred at the same temperature for 6 hours under a nitrogen atmosphere. Water (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (60 mL). The organic layer was washed three times with sodium sulfite solution (200 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give intermediate S2 (4.1 g, 65.09% yield). MS: m / z = 502.1 (M+1, ESI+).

[0286] Preparation of intermediate S3: N-benzyl-N-(6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester

[0287]

[0288] Step 1: Preparation of tert-butyl N-benzyl-N-(3-iodo-6-morpholinoimidazo[l,2-b]pyridazin- 8-yl)glycinate Step 2: Preparation of tert-butyl N-benzyl-N-(6-morpholino-3-(trifluoromethyl)imidazo[l,2- b]pyridazin-8-yl)glycinate

[0289] Intermediate S1 (5.1 g, 11.34 mmol) was dissolved in THF (80 mL), and then NIS (3.72 g, 16.53 mmol) was added at 0 °C and stirred for 2 hours. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (400 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give N-benzyl-N-(3-iodo-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (5.99 g, yield 72.13%). MS: m / z = 550.1 (M+1, ESI+).

[0290] Step 1: Preparation of tert-butyl (6-chloroimidazo[l,2-b]pyridazin-8-yl)glycinate Step 2: Preparation of tert-butyl N-(6-chloroimidazo[l,2-b]pyridazin-8-yl)-N-(4- methoxybenzyl)glycinate

[0291] The product obtained in step 1 (5.95 g, 10.83 mmol) and methyl 2,2-difluoro-2-(fluorosulfonyl)acetate (20.81 g, 108.30 mmol) were dissolved in DMF (150 mL), and then CuI (10.31 g, 54.15 mmol) was added and the mixture was stirred at 100 °C for 16 hours. Water (1000 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (300 mL). The organic layer was washed three times with brine (800 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give intermediate S3 (1.4 g, yield 26.30%). MS: m / z = 492.2 (M+1, ESI+).

[0292] Preparation of intermediate S4: N-(6-chloroimidazolo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine tert-butyl ester

[0293]

[0294] Step 1: Preparation of tert-butyl N-(4-methoxybenzyl)-N-(6-morpholinoimidazo[l,2-b]pyridazin- 8-yl)glycinate

[0295] 8-Bromo-6-chloro-imidazo[1,2-b]pyridazine (100 g, 430.17 mmol) was dissolved in glycine tert-butyl ester (1000 mL), and the mixture was stirred at 120 °C for 16 hours. Water (3000 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (1000 mL). The organic layer was washed three times with brine (3000 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give (6-chloroimidazo[1,2-b]pyridazine-8-yl)glycine tert-butyl ester (102 g, 83.87% yield). MS: m / z = 283.1 (M+1, ESI+).

[0296] Step 2: Preparation of tert-butyl N-(3-bromo-6-morpholinoimidazo[l,2-b]pyridazin-8-yl)-N- (4-methoxybenzyl)glycinate Step 1: Preparation of 6-chloro-3-iodo-N,N-bis(4-methoxybenzyl)imidazo[l,2-b]pyridazin-8- amine

[0297] The product obtained in step 1 (102 g, 361.7 mmol) and 4-methoxybenzyl chloride (62.1 g, 397.9 mmol) were dissolved in MeCN (1000 mL), followed by the addition of cesium carbonate (178 g, 542.6 mmol) and stirring at 60 °C for 16 hours. Water (5000 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (1000 mL). The organic layer was washed three times with brine (5000 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give intermediate S4 (110.5 g, 76% yield). MS: m / z = 403.1 (M+1, ESI+).

[0298] Preparation of intermediate S5: N-(3-bromo-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine tert-butyl ester

[0299]

[0300] Step 2: Preparation of 6-chloro-N,N-bis(4-methoxybenzyl)-3-(trifluoromethyl)imidazo[l,2- b]pyridiazin-8-amine Step 1: Preparation of 6-chloro-N-(4-methoxybenzyl)-3-(trifluoromethyl)imidazo [l,2-b]pyridazin- 8-amine

[0301] Intermediate S4 (15 g, 37.23 mmol) and morpholine (33.1 mL, 372.33 mmol) were dissolved in 1,4-dioxane (300 mL), followed by the addition of cesium carbonate (37 g, 112 mmol) and RuPhos-Pd-G3 (3.12 g, 3.72 mmol) and stirring at 110 °C for 16 hours. Water (800 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (300 mL). The organic layer was washed three times with brine (800 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give N-(4-methoxybenzyl)-N-(6-morpholinylimidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (15.2 g, 90.01% yield). MS: m / z = 454.2 (M+1, ESI+).

[0302] Step 2: Preparation of N-(6-chloro-3-(trifluoromethyl)imidazo[l,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine ethyl ester Step 1: Preparation of N,N-bis(4-methoxybenzyl)-6-morpholino-3-(trifluoromethyl)imidazo[l, 2-b]pyridazin-8-amine

[0303] The product obtained in step 1 (15.2 g, 33.51 mmol) was dissolved in THF (200 mL), and then NBS (5.96 g, 33.51 mmol) was added at -60 °C, and the mixture was stirred at this temperature for 1 hour under a nitrogen atmosphere. Water (800 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (200 mL). The organic layer was washed three times with sodium sulfite solution (800 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give intermediate S5 (12 g, yield 67.25%). MS: m / z = 532.1 (M+1, ESI+).

[0304] Preparation of intermediate S6: 6-chloro-N,N-bis(4-methoxybenzyl)imidazo[1,2-b]pyridazine-8-amine

[0305]

[0306] 8-Bromo-6-chloroimidazolo[1,2-b]pyridazine (90 g, 390 mmol) and bis(4-methoxybenzyl)amine (120 g, 468 mmol) were dissolved in DMF (800 mL), and then DIEA (135.6 mL, 780 mmol) was added and the mixture was stirred overnight at 90 °C. Water (4000 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (800 mL). The organic layer was washed three times with brine (4000 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give intermediate S6 (120 g, 75.95% yield). MS: m / z = 409.1 (M+1, ESI+).

[0307] Preparation of intermediate S7: 6-chloro-N,N-bis(4-methoxybenzyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0308]

[0309] Step 2: Preparation of N-(4-methoxybenzyl)-6-morpholino-3-(trifluoro methyl)imidazo[l,2-b]pyridazin-8-amine

[0310] Intermediate S6 (120 g, 294 mmol) was dissolved in DMF (800 mL), and then NIS (66 g, 294 mmol) was added and stirred at 60 °C for 16 hours. Sodium thiosulfate solution (8000 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (1000 mL). The organic layer was washed three times with brine (8000 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give 6-chloro-3-iodo-N,N-bis(4-methoxybenzyl)imidazo[1,2-b]pyridazin-8-amine (139 g, 88.53% yield). MS: m / z = 535.1 (M+1, ESI+).

[0311] Step 3: Preparation of N-(4-methoxybenzyl)-N-(6-morpholino-3-(trifluoromethyl)imidazo[l,2- b]pyridazin-8-yl)glycine ethyl ester Step 4: Preparation of N-(4-methoxybenzyl)-N-(6-morphol ino-3-(trifluoromethyl)imidazo[l,2- b]pyri dazin-8-yl)glycine

[0312] The product obtained in step 1 (139 g, 260.3 mmol) and methyl 2,2-difluoro-2-(fluorosulfonyl)acetate (250 g, 1.3 mol) were dissolved in DMF (1000 mL), followed by the addition of CuI (123.6 g, 650.8 mmol) and DIEA (226 mL, 1.3 mol) and stirring at 85 °C for 16 hours. Water (8000 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (2000 mL). The organic layer was washed three times with brine (8000 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give intermediate S7 (106 g, 85.48% yield). MS: m / z = 476.1 (M+1, ESI+).

[0313] Preparation of intermediate S8: N-(6-chloro-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine ethyl ester

[0314]

[0315] Step 1: Preparation of tert-butyl 3-(8-((2-ethoxy-2-oxoethyl)(4-methoxybenzyl)amino)-3- (trifluoromethyl)imidazo[l,2-b]pyridazin-6-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate

[0316] Intermediate S7 (106 g, 222.7 mmol) was dissolved in DCM (600 mL), then TFA (200 mL) was added and the mixture was stirred at 25 °C for 1 hour. Sodium bicarbonate solution (3000 mL) was added to the reaction mixture, and the product was extracted three times with DCM (500 mL). The organic layer was washed three times with brine (3000 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give 6-chloro-N-(4-methoxybenzyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (72 g, 90.8% yield). MS: m / z = 357.2 (M+1, ESI+).

[0317] Step 2: Preparation of (6-(3,6-diazabicyclo[3.1.1]heptan-3-yl)-3-(trifluoromethyl)imidazo[l,2- b] pyridazin-8-yl)methanol ​

[0318] The product obtained in step 1 (29 g, 81.5 mmol) and ethyl 2-bromoacetate (20.4 g, 122.2 mmol) were dissolved in THF (200 mL), followed by the addition of cesium carbonate (53.14 g, 163 mmol) and stirring at 70 °C for 16 hours. Water (800 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (200 mL). The organic layer was washed three times with brine (800 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give intermediate S8 (27 g, 75% yield). MS: m / z = 443.1 (M+1, ESI+).

[0319] Preparation of intermediate S9: N-(4-methoxybenzyl)-N-(6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycine

[0320]

[0321] ​ ​

[0322] Morpholine (120 mL) was added to intermediate S7 (12 g, 25.2 mmol), and the mixture was stirred at 120 °C for 16 hours. After cooling and concentrating the reaction mixture, it was purified by silica gel chromatography to give N,N-bis(4-methoxybenzyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine (5 g, yield 37.23%). MS: m / z = 527.3 (M+1, ESI+).

[0323] ​ ​

[0324] The product obtained in step 1 (36 g, 68.31 mmol) was dissolved in DCM (300 mL), and then TFA (100 mL) was added. The mixture was stirred at 25 °C for 1 hour. Sodium bicarbonate solution (1000 mL) was added to the reaction mixture, and the product was extracted three times with DCM (100 mL). The organic layer was washed three times with brine (1000 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(4-methoxybenzyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine (27 g, 97.12% yield). MS: m / z = 408.2 (M+1, ESI+).

[0325] ​ ​

[0326] The product obtained in step 2 (8.4 g, 20.62 mmol) was dissolved in DMF (100 mL), and then NaH (1.24 g, 30.93 mmol, 60% purity) was added in portions at 0 °C and stirred for 30 min. Ethyl 2-bromoacetate (5.17 g, 30.93 mmol) was added to the reaction mixture, and the mixture was stirred at 25 °C for 2 h. Water (800 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (200 mL). The organic layer was washed three times with brine (800 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give N-(4-methoxybenzyl)-N-(6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycine ethyl ester (8.2 g, 80.59% yield). MS: m / z = 494.2 (M+1, ESI+).

[0327] ​ ​

[0328] The product obtained in step 3 (8.2 g, 16.62 mmol) was dissolved in 1,4-dioxane (80 mL) and water (20 mL), followed by the addition of sodium hydroxide (1.6 g, 40 mmol), and the mixture was stirred at 100 °C for 1 hour. 1 N hydrochloric acid (500 mL) was added to the reaction mixture, and the product was extracted three times with DCM (150 mL). The organic layer was washed three times with brine (500 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give intermediate S9 (7 g, 90.51% yield). MS: m / z = 466.1 (M+1, ESI+).

[0329] Preparation of intermediate S10: N-(4-methoxybenzyl)-N-(6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycyl chloride

[0330]

[0331] Intermediate S9 (600 mg, 1.29 mmol) was dissolved in DMF (10 μL) and DCM (6 mL), and then oxalyl chloride ((COCl)2, 1.42 mmol, 0.12 mL) was added, and the mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated to give intermediate S10 (600 mg, crude product) as a yellow solid.

[0332] Preparation of intermediate S11: (6-(6-methyl-3,6-diazabicyclo[3.1.1]heptane-3-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycine ethyl ester

[0333]

[0334] ​ ​

[0335] Intermediate S8 (8.5 g, 19.23 mmol) and intermediate A44 (11.42 g, 57.69 mmol) were dissolved in 1,4-dioxane (100 mL), followed by the addition of RuPhos-Pd-G3 (1.6 g, 1.92 mmol), RuPhos (1.8 g, 3.84 mmol), and cesium carbonate (18.75 g, 57.69 mmol). The mixture was stirred at 110 °C for 48 hours under a nitrogen atmosphere. Water (500 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (100 mL). The organic layer was washed three times with brine (500 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give 1.2 g (10.34% yield) of tert-butyl 3-(8-((2-ethoxy-2-oxoethyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate as a yellow solid. MS: m / z = 605.3 (M+1, ESI+).

[0336] ​ Preparation of ethyl (6-(6-methyl-3,6-diazabicyclo[3.1.1]octan-3-yl)-3- (trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycinate

[0337] The product obtained in step 1 (1.2 g, 1.98 mmol) was dissolved in DCM (18 mL), and then TFA (6 mL) was added. The mixture was stirred at 25 °C for 2 hours. Sodium bicarbonate aqueous solution (200 mL) was added to the reaction mixture, and the product was extracted three times with DCM (50 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give ethyl glycine (600 mg, 78.95% yield) as a yellow solid (6-(3,6-diazabicyclo[3.1.1]heptane-3-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycine. MS: m / z = 385.0 (M+1, ESI+).

[0338] Step 3: Preparation of ethyl (6-(6-methyl-3,6-diazabicyclo[3.1.1]octan-3-yl)-3- (trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycine Step 1: Preparation of tert-butyl 3-methyl-3,8-diazabicyclo[3.2.1]octane-8- carboxylate

[0339] The product obtained in step 2 (600 mg, 1.56 mmol) was dissolved in THF (12 mL), then formaldehyde (163.8 mg, 1.638 mmol) and AcOH (2 drops) were added and stirred at 25 °C for 0.5 h. NaBH(OAc)3 (494 mg, 2.34 mmol) was then added and stirred at 25 °C for 16 h. Water (100 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (30 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give intermediate S11 (500 mg, 80.26% yield) as a yellow solid. MS: m / z = 399.1 (M+1, ESI+).

[0340] Preparation of intermediate S12: (6-(3-methyl-3,6-diazabicyclo[3.1.1]heptane-6-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycine ethyl ester

[0341]

[0342] Intermediate S12 (220 mg, 64.32% yield), a yellow solid, was obtained using the same method as described in steps 1 to 3 for the preparation of intermediate S11, except that intermediate A45 (1.88 g, 9.48 mmol) was used instead of intermediate A44 (11.42 g, 57.69 mmol) in step 1. MS: m / z = 399.1 (M+1, ESI+).

[0343] Preparation of intermediate A7: 3-methyl-3,8-diazabicyclo[3.2.1]octane-8-onium trifluoroacetate

[0344]

[0345] Step 2: Preparation of 3-methyl-3,8-diazabicyclo[3.2.1 ]octan-8-ium trifluoroacetate

[0346] 3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (7 g, 33.13 mmol) and formaldehyde (38.15 g, 508.23 mmol, 35.00 mL, 40% purity) were dissolved in methanol (50 mL) and THF (50 mL), and then AcOH (2.14 g, 16.56 mmol) was added and the mixture was stirred at 25 °C for 1 hour. NaBH(OAc)3 (10.53 g, 49.69 mmol) was added to the reaction mixture and the mixture was stirred at 25 °C for 16 hours. The reaction mixture was concentrated and the concentrate was purified by silica gel chromatography to give 3-methyl-3,8-diazabicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (6.6 g, yield 88.42%).

[0347] Step 1: Preparation of 8-benzyl-3-methyl-8-azabicyclo[3.2.1]octan-3-ol

[0348] The product obtained in step 1 (7.6 g, 33.58 mmol) was dissolved in DCM (30 mL), and then TFA (12.9 mL, 167.91 mmol) was added and the mixture was stirred at 25 °C for 16 hours. The reaction mixture was concentrated to give intermediate A7 (9.4 g, crude product).

[0349] Preparation of intermediate A16: Azacyclobutane-3-carboxynitrile trifluoroacetate

[0350]

[0351] 3-Cyanozycyclobutane-1-carboxylic acid tert-butyl ester (3 g, 16.46 mmol) was dissolved in THF (30 mL), then TFA (10 mL) was added and the mixture was stirred at 25 °C for 16 hours. The reaction mixture was concentrated to give intermediate A16 (1.8 g, crude). MS: m / z = 83.2 (M+1, ESI+).

[0352] Preparation of intermediate A20: 3-methyl-8-azabicyclo[3.2.1]octane-3-ol

[0353]

[0354] Step 2: Preparation of 3-methyl-8-azabicyclo[3.2.1]octan -3-ol

[0355] 10.00 g (46.45 mmol) of 8-benzyl-8-azabicyclo[3.2.1]octane-3-one was dissolved in 100 mL of THF, and then 3 M methyl magnesium bromide solution (39 mL) was slowly added over 30 minutes at 0 °C, while the mixture was stirred at 25 °C for 3 hours. Ice water (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (60 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give 6 g (55.84%) of 8-benzyl-3-methyl-8-azabicyclo[3.2.1]octane-3-ol. MS: m / z = 232.2 (M+1, ESI+).

[0356] Step 1: Preparation of methyl 1-benzylazetidine-3-carboxylate

[0357] The product obtained in step 1 (6 g, 25.94 mmol) was dissolved in methanol (50 mL), then Pd / C (1 g) was added and the mixture was stirred at 25 °C for 16 hours under a N2 atmosphere. The reaction mixture was filtered and concentrated to give intermediate A20 (3 g, yield 81.91%). MS: m / z = 142.1 (M+1, ESI+).

[0358] Preparation of intermediate A26: 4-cyanopiperidine-1-onium trifluoroacetate

[0359]

[0360] 3 g of tert-butyl 4-cyanopiperidine-1-carboxylate (14.27 mmol) was dissolved in 20 mL of THF, and then 5 mL of TFA was added and the mixture was stirred at 25 °C for 16 hours. The reaction mixture was concentrated to give intermediate A26 (2.8 g, crude product).

[0361] Preparation of intermediate A29: 2-azaspiro[3.3]heptane-6-ol trifluoroacetate

[0362]

[0363] 4 g (18.76 mmol) of tert-butyl 6-hydroxy-2-azaspiro[3.3]heptane-2-carboxylate was dissolved in DCM (30 mL), followed by the addition of TFA (10 mL), and the mixture was stirred at 25 °C for 16 hours. The reaction mixture was concentrated to give intermediate A29 (2.5 g, crude). MS: m / z = 113.9 (M+1, ESI+).

[0364] Preparation of intermediate A31: pyrrolidine-3-carboxynitrile trifluoroacetate

[0365]

[0366] 3-Cyanopyrrolidine-1-carboxylic acid tert-butyl ester (1.8 g, 9.17 mmol) was dissolved in DCM (15 mL), then TFA (3 mL) was added, and the mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated to give intermediate A31 (1.12 g, crude product). MS: m / z = 97.1 (M+1, ESI+).

[0367] Preparation of intermediate A32: 1-(azacyclobutan-3-yl)cyclopropan-1-ol

[0368]

[0369] Step 2: Preparation of 1-(1-benzylazetidin-3-yl)cyclopropan-1-ol

[0370] Methyl aziridine-3-carboxylate hydrochloride (20 g, 132 mmol) and BnBr (17.22 mL, 145 mmol) were dissolved in DMF (300 mL), and then DIEA (115 mL, 660 mmol) was added in portions. The mixture was stirred at 25 °C for 16 hours. Ammonium chloride aqueous solution (2000 mL) was added to the reaction mixture, and the mixture was extracted three times with EtOAc (400 mL). The organic layer was washed three times with brine (1000 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give methyl 1-benzylaziridine-3-carboxylate (22 g, 81.24% yield). MS: m / z = 205.9 (M+1, ESI+).

[0371] Step 3: Preparation of 1-(azetidin-3-yl)cyclopropan-1-ol

[0372] The product obtained in step 1 (16 g, 77.95 mmol) was dissolved in THF (200 mL), and then Ti(i-PrO)4 (26.59 g, 93.54 mmol) was added. A 1 M solution of ethyl magnesium bromide (311.81 mL) was slowly added over 30 minutes at -60 °C under an Ar atmosphere. The reaction mixture was heated to room temperature and stirred for 16 hours. An aqueous solution of ammonium chloride (1000 mL) was added to the reaction mixture, and the mixture was extracted three times with EtOAc (300 mL). The organic layer was washed with brine (100 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give 1-(1-benzylazetane-3-yl)cyclopropane-1-ol (9.5 g, yield 59.95%). MS: m / z = 204.2 (M+1, ESI+).

[0373] Step1: Preparation of tert-butyl 3-(((benzyloxy)carbonyl)(cyclopropyl)amino)azetidine- 1-carboxylate

[0374] The product obtained in step 2 (9.5 g, 46.73 mmol) was dissolved in methanol (120 mL), and then Pd / C (4.5 g, 10% purity) was added. The mixture was stirred at 60 °C for 16 hours under a H2 atmosphere. The reaction mixture was filtered and concentrated to give intermediate A32 (4.1 g, 77.53% yield). 1 H NMR (400 MHz, MeOD) δ 4.13 (dd, 4H), 2.76-2.62 (m, 1H), 0.79-0.76 (m, 2H), 0.54-0.47 (m, 2H); MS: m / z = 113.9 (M+1,ESI+).

[0375] Preparation of intermediate A36: Azacyclobutane-3-yl(cyclopropyl)carbamate benzyl ester

[0376]

[0377] Step 2: Preparation of benzyl azetidin-3-yl(cyclopropyl)carbamate

[0378] 3-(cyclopropylamino)azacyclobutane-1-carboxylic acid tert-butyl ester (5 g, 23.55 mmol) and CbzOSu (8.80 g, 35.33 mmol) were dissolved in DCM (80 mL), followed by the addition of TEA (9.85 mL, 70.66 mmol) and stirring at 25 °C for 16 hours. Water (200 mL) was added to the reaction mixture, and the product was extracted three times with DCM (60 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give 3-(((benzyloxy)carbonyl)(cyclopropyl)amino)azacyclobutane-1-carboxylic acid tert-butyl ester (7 g, yield 85.79%). MS: m / z = 347.1 (M+1, ESI+).

[0379] Step 1: Preparation of tert-butyl 3-(((tert-butyldiphenylsilyl)oxy)methyl)azetidine-1- carboxylate

[0380] The product obtained in step 1 (7 g, 20.21 mmol) was dissolved in DCM (60 mL), then TFA (20 mL) was added and the mixture was stirred at 25 °C for 16 hours. After concentrating the reaction mixture, the pH was adjusted to 8–9, and the mixture was extracted three times with DCM (80 mL). The organic layer was washed three times with brine (200 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give intermediate A36 (4 g, 80.37% yield). MS: m / z = 247.2 (M+1, ESI+).

[0381] Preparation of intermediate A37: 3-((tert-butyldimethylsilyl)oxy)azacyclobutane

[0382]

[0383] Azacyclobutane-3-ol hydrochloride (5 g, 46 mmol) and TBDMSCl (13.8 g, 92 mmol) were dissolved in DCM (50 mL), and then TEA (19.10 mL, 137 mmol) was added and the mixture was stirred at 25 °C for 16 hours. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with DCM (80 mL). The organic layer was washed three times with brine (800 mL) and three times with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give intermediate A37 (5.8 g, yield 67.83%). MS: m / z = 188.2 (M+1, ESI+).

[0384] Preparation of intermediate A38: 3-(((tert-butyldiphenylsilyl)oxy)methyl)azacyclobutane trifluoroacetate

[0385]

[0386] Step 2: Preparation of 3-(((tert-butyldiphenylsilyl)oxy)methyl)-azetidine trifluoroacetate Step 1: Preparation of benzyl 8-azaspiro[bicyclo[3.2.1]octane-3,2'-[1,3]dioxolane]-8- carboxylate

[0387] 3-(hydroxymethyl)azacyclobutane-1-carboxylate (5 g, 26.70 mmol) and TBDPSCl (4.66 g, 40.06 mmol) were dissolved in DCM (80 mL), followed by the addition of DMAP (326 mg, 2.67 mmol) and imidazole (2.73 g, 40.06 mmol) and stirring at 25 °C for 2 h. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with DCM (80 mL). The organic layer was washed three times with brine (300 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give tert-butyl 3-(((tert-butyldiphenylsilyl)oxy)methyl)azacyclobutane-1-carboxylate (8 g, 70.38%). MS: m / z = 426.0 (M+1, ESI+).

[0388] Step 2: Preparation of 8-azaspiro[bicyclo[3.2.1]octane -3,2'-[1,3]dioxolane]

[0389] The product obtained in step 1 (8 g, 18.80 mmol) was dissolved in DCM (60 mL), and then TFA (20 mL) was added and stirred at 25 °C for 16 hours. After concentrating the reaction mixture, intermediate A38 (6 g, crude product) was obtained. MS: m / z = 325.0 (M+1, ESI+).

[0390] Preparation of intermediate A40: (R)-3-((tert-butyldimethylsilyl)oxy)pyrrolidine

[0391]

[0392] Intermediate A40 (6.2 g, 76.10% yield) was prepared using (R)-pyrrolidine-3-ol hydrochloride (5 g, 40.46 mmol) as the starting material, following the method described for intermediate A37. MS: m / z = 202.0 (M+1, ESI+).

[0393] Preparation of intermediate A41: (S)-3-((tert-butyldimethylsilyl)oxy)pyrrolidine

[0394]

[0395] Using (S)-pyrrolidine-3-ol hydrochloride (11 g, 126 mmol) as the starting material, intermediate A41 (15 g, 58.99% yield) was obtained by the same method described for the preparation of intermediate A37. MS: m / z = 202.2 (M+1, ESI+).

[0396] Preparation of intermediate A47: 8-azaspiro[bicyclo[3.2.1]octane-3,2'-[1,3]dioxolane]

[0397]

[0398] Step 1: Preparation of 4-chloro-5-methoxy-2-nitroaniline

[0399] 3-oxo-8-azabicyclo[3.2.1]octane-8-carboxylic acid benzyl ester (10 g, 38.61 mmol) was dissolved in toluene (60 mL), followed by the addition of TsOH (664 mg, 3.86 mmol) and ethylene glycol (11.97 g, 193.05 mmol) and stirring at 120 °C for 48 hours. Water (500 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (100 mL). The organic layer was washed three times with brine (500 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give 8-azaspiro[bicyclo[3.2.1]octane-3,2'-[1,3]dioxolane]-8-carboxylic acid benzyl ester (6.8 g, yield 58.27%) as a yellow solid. MS: m / z = 304.1 (M+1, ESI+).

[0400] Step 2: Preparation of 4-chloro-5-methoxybenzene-1,2-diamine

[0401] The product obtained in step 1 (6.8 g, 22.44 mmol) was dissolved in methanol (50 mL), and then Pd / C (1 g) was added and the mixture was stirred at 25 °C for 16 hours under a H2 atmosphere. The reaction mixture was concentrated and filtered, and the concentrate was purified by silica gel chromatography to give intermediate A47 (3.7 g, 97.63% yield) as a yellow solid. MS: m / z = 170.1 (M+1, ESI+).

[0402] Preparation of intermediate B6: 4-chloro-5-methoxyphenyl-1,2-diamine

[0403]

[0404] Step 1: Preparation of 5-methyl-2-nitro-4-(trifluoromethyl)aniline

[0405] 4,5-Dichloro-2-nitroaniline (4 g, 19.32 mmol) was dissolved in methanol (25 mL), and sodium methoxide (1.05 g, 19.32 mmol) was added. The mixture was stirred at 70 °C for 4 hours. Ice water (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give 4-chloro-5-methoxy-2-nitroaniline (3.4 g, 86.85% yield). MS: m / z = 203.1 (M+1, ESI+).

[0406] Step 2: Preparation of 4-methyl-5-(trifluoromethyl)benzene-1,2-diamine

[0407] The product obtained in step 1 (3.4 g, 16.78 mmol) was dissolved in ethanol (50 mL), and then ammonium chloride (13.5 g, 252 mmol), zinc (16.4 g, 252 mmol), and formic acid (1.98 mL, 50.4 mmol) were added at 0 °C, and the mixture was stirred at 25 °C for 2 hours. The reaction mixture was filtered and concentrated, and then ice water (200 mL) was added. The product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give intermediate B6 (2.3 g, yield 79.40%). MS: m / z = 173.1 (M+1, ESI+).

[0408] Preparation of intermediate B15: 4-methyl-5-(trifluoromethyl)phenyl-1,2-diamine

[0409]

[0410] Step 1: Preparation of 1-iodo-4-nitro-2-(trifluoromethyl)benzene

[0411] 5-Chloro-2-nitro-4-(trifluoromethyl)aniline (10 g, 41.57 mmol) was dissolved in dimethyl sulfoxide (30 mL), followed by the addition of diethyl malonate (6.65 g, 41.57 mmol) and potassium tert-butoxide (11.66 g, 103.92 mmol), and the mixture was stirred at 60 °C for 16 hours. After cooling the reaction solution to room temperature, potassium hydroxide (10.96 g, 195 mmol) and water (30 mL) were added, and the mixture was stirred again at 60 °C for 16 hours. Water (400 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (400 mL), dried over Na₂SO₄, filtered, concentrated, and then purified by silica gel chromatography to give 5-methyl-2-nitro-4-(trifluoromethyl)aniline (5.97 g, 65.24% yield). MS: m / z = 220.2 (M+1, ESI+).

[0412] Step 2: Preparation of 4-nitro-1,2-bis(trifluoromethyl)benzene

[0413] The product obtained in step 1 (5.97 g, 27.12 mmol) was dissolved in methanol (20 mL), followed by the addition of sodium bisulfite (23.61 g, 135.59 mmol) and sodium hydroxide (3.25 g, 81.35 mmol), and the mixture was stirred at 25 °C for 16 hours. Water (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (50 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give intermediate B15 (2.13 g, yield 41.30%). MS: m / z = 191.1 (M+1, ESI+).

[0414] Preparation of intermediate B16: 4,5-bis(trifluoromethyl)phenyl-1,2-diamine

[0415]

[0416] Step 3: Preparation of 3,4-bis(trifluoromethyl)aniline

[0417] 4-Nitro-2-(trifluoromethyl)aniline (50 g, 242 mmol) was dissolved in water (600 mL) and sulfuric acid (400 mL) at 0 °C, followed by the slow addition of sodium nitrite (33.48 g, 484 mmol) dissolved in water (160 mL). After stirring at the same temperature for 1 hour, potassium iodide (KI, 181.8 g, 1212 mmol) dissolved in water (1000 mL) was added, and the mixture was stirred at 25 °C for 16 hours. The reaction mixture was filtered, washed, and concentrated to give 1-iodo-4-nitro-2-(trifluoromethyl)benzene (74 g, 96.17% yield). 1 H NMR (400 MHz, CDCl3) δ 8.48 (d, 1H), 8.28 (d, 1H), 8.05 (dd, 1H).

[0418] Step 4: Preparation of N-(3,4-bis(trifluoromethyl)phenyl)-2,2,2-trifluoroacetamide

[0419] The product obtained in step 1 (74 g, 233.4 mmol) and methyl 2,2-difluoro-2-(fluorosulfonyl)acetate (Chen's reagent, 224.2 g, 1167.2 mmol) were dissolved in DMF (800 mL), and then CuI (44.4 g, 233.4 mmol) was added and the mixture was stirred at 80 °C for 16 hours under N2 atmosphere. Water (8000 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (2000 mL). The organic layer was washed three times with brine (8000 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give 4-nitro-1,2-bis(trifluoromethyl)benzene (26 g, yield 42.97%).

[0420] Step 5: Preparation of 2,2,2-trifluoro-N-(2-nitro-4,5-bis(trifluoromethyl)phenyl)acetamide

[0421] The product obtained in step 2 (26 g, 100.4 mmol) and stannous chloride (SnCl2, 114.2 g, 602.1 mmol) were dissolved in ethanol (2000 mL), and the mixture was stirred at 70 °C for 16 hours under a nitrogen atmosphere. Water (800 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (200 mL). The organic layer was washed three times with brine (800 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give 3,4-bis(trifluoromethyl)aniline (21 g, 91.3% yield).

[0422] Step 6: Preparation of 2-nitro-4,5-bis(trifluoromethyl)aniline

[0423] The product obtained in step 3 (21.2 g, 92.54 mmol) was dissolved in 1,4-dioxane (150 mL), and then trifluoroacetic anhydride (TFAA, 38.88 g, 185.05 mmol, 25.73 mL) was added and the mixture was stirred at 25 °C for 16 hours. Water (1000 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (300 mL). The organic layer was washed three times with brine (1000 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give N-(3,4-bis(trifluoromethyl)phenyl)-2,2,2-trifluoroacetamide (21.7 g, yield 72.14%). MS: m / z = 324.1 (M-1, ESI-).

[0424] Step 7: Preparation of 4,5-bis(trifluoromethyl)benzene-1,2-diamine

[0425] The product obtained in step 4 (21.6 g, 66.31 mmol) was dissolved in sulfuric acid (200 mL), and then potassium nitrate (13.42 g, 132.62 mmol) was added at 0 °C, and the mixture was stirred at 25 °C for 16 hours. Ice water (2000 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (400 mL). The organic layer was washed three times with brine (2000 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give 2,2,2-trifluoro-N-(2-nitro-4,5-bis(trifluoromethyl)phenyl)acetamide (12.5 g, yield 50.83%). MS: m / z = 369.0 (M⁻¹, ESI⁻).

[0426] Step 2: Preparation of 5,6-dichloro-1-(phenylsulfonyl)-1H-indole-2-carbaldehyde

[0427] The product obtained in step 5 (12.5 g, 33.77 mmol) was dissolved in methanol (150 mL), and then potassium carbonate (23.3 g, 168.85 mmol) was added and the mixture was stirred at 25 °C for 16 hours. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give 2-nitro-4,5-bis(trifluoromethyl)aniline (10.4 g, crude). MS: m / z = 273.0 (M-1, ESI-).

[0428] Step 1: Preparation of 1-((2-(trimethylsilyl)ethoxy)methyl)-4,5,6,7-tetrahydro-1H- benzo[d]imidazole

[0429] The product obtained in step 6 (10.4 g, 37.96 mmol) and stannous chloride (SnCl2, 43.18 g, 227.74 mmol) were dissolved in ethanol (100 mL), and the mixture was stirred at 70 °C for 16 hours under a nitrogen atmosphere. Water (800 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (200 mL). The organic layer was washed three times with brine (800 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give intermediate B16 (8.5 g, 94.55% yield). MS: m / z = 243.0 (M-1, ESI-).

[0430] Preparation of intermediate B20: 5,6-dichloro-1-(benzenesulfonyl)-1H-indole-2-carboxaldehyde

[0431]

[0432] ​

[0433] 5,6-Dichloro-1H-indole (3 g, 16.13 mmol) was dissolved in DMF (30 mL), and then NaH (774 mg, 19.35 mmol, 60% purity) was added in portions at 0 °C, with stirring at the same temperature for 1 hour. Subsequently, benzenesulfonyl chloride (4.27 g, 24.19 mmol) was added, and stirring was performed at room temperature for 2 hours. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (60 mL). The organic layer was washed three times with brine (300 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give 5,6-dichloro-1-(benzenesulfonyl)-1H-indole (4.5 g, 85.56% yield) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.13 (s, 1H), 8.09-8.03 (m,2H), 7.97 (d, 1H), 7.93 (s, 1H), 7.74 (t, 1H), 7.64 (t, 2H), 6.87 (d, 1H).

[0434] ​

[0435] The product obtained in step 1 (4.5 g, 13.80 mmol) was dissolved in THF (50 mL), and then a 2 M solution of diisopropylaminolithium (2 M LDA, 13.8 mL, 27.6 mmol) was slowly added at -60 °C, and the mixture was stirred at the same temperature for 1 hour. Subsequently, DMF (2.02 g, 27.59 mmol, 1.42 mL) was added, and the mixture was stirred at -60 °C for 2 hours. Water (200 mL) was added to the reaction mixture, and the mixture was extracted three times with EtOAc (40 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give intermediate B20 (3 g, 61.39% yield) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.12 (s, 1H), 8.05 (d, 2H),7.98-7.93 (m, 2H), 7.74 (t, 1H), 7.63 (t, 2H), 6.86 (d, 1H);MS: m / z = 354.0(M+1, ESI+).

[0436] Preparation of intermediate B22: 5,6-Dimethyl-1-(benzenesulfonyl)-1H-indole-2-carboxaldehyde

[0437]

[0438] Intermediate B22 (1.3 g, yield 33.82%), a yellow solid, was obtained by using the same method as described in steps 1 and 2 for the preparation of intermediate B20, except that 5,6-dimethyl-1H-indole (2 g, 13.77 mmol) was used instead of 5,6-dichloro-1H-indole (3 g, 16.13 mmol) in step 1.

[0439] 1 H NMR (400 MHz, DMSO-d6) δ 10.32 (s, 1H), 7.94 (s, 1H), 7.92-7.87 (m,2H), 7.70 (t, 1H), 7.61-7.55 (m, 3H), 7.52 (s, 1H), 2.41 (s, 3H), 2.28 (s,3H); MS: m / z = 314.1 (M+1, ESI+).

[0440] Preparation of intermediate B23: 4-fluoro-1-(benzenesulfonyl)-1H-indole-2-carboxaldehyde

[0441]

[0442] Intermediate B23 (3.9 g, yield 64.36%), a yellow oily substance, was obtained by using the same method as described in steps 1 and 2 for preparing intermediate B20, except that 4-fluoro-1H-indole (5.1 g, 37.74 mmol) was used instead of 5,6-dichloro-1H-indole (3 g, 16.13 mmol) in step 1.

[0443] Preparation of intermediate B24: 4,7-difluoro-1-(benzenesulfonyl)-1H-indole-2-carboxaldehyde

[0444]

[0445] Intermediate B24 (3 g, 78.24% yield), a yellow oily substance, was obtained using the same methods described in steps 1 and 2 for the preparation of intermediate B20, except that 4,7-difluoro-1H-indole (2 g, 13.06 mmol) was used instead of 5,6-dichloro-1H-indole (3 g, 16.13 mmol) in step 1. MS: m / z = 322.1 (M+1, ESI+).

[0446] Preparation of intermediate B25: 4,5,6,7-tetrahydro-1H-indole-2-carboxaldehyde

[0447]

[0448] 4,5,6,7-Tetrahydro-1H-indole (2 g, 16.50 mmol) was dissolved in DMF (20 mL), and then phosphoryl chloride (POCl3, 2.46 mL, 26.41 mmol) was added in portions at -20 °C and stirred at room temperature for 2 hours. Water (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (60 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give intermediate B25 (1.9 g, 77.16% yield). 1 H NMR (400 MHz, DMSO-d6) δ 11.61 (s, 1H), 9.27 (s, 1H), 6.68 (d, 1H), 2.56 (t, 2H), 2.45 (t, 2H), 1.75-1.65 (m, 4H); MS: m / z = 150.1 (M+1,ESI+).

[0449] Preparation of intermediate B26: 7-hydroxy-1H-indole-2-carboxaldehyde

[0450]

[0451] Using the same method as that used to prepare intermediate B25, 1H-indole-7-ol (3 g, 22.53 mmol) was used instead of 4,5,6,7-tetrahydro-1H-indole (2 g, 16.50 mmol) to give intermediate B26 (3.1 g, 85.37% yield) as a red solid. MS: m / z = 162.2 (M+1, ESI+).

[0452] Preparation of intermediate B27: 6-hydroxy-1H-indole-2-carboxaldehyde

[0453]

[0454] Using the same method as that used to prepare intermediate B25, 1H-indole-6-ol (3 g, 22.53 mmol) was used instead of 4,5,6,7-tetrahydro-1H-indole (2 g, 16.50 mmol) to give intermediate B27 (3 g, 82.62% yield) as a yellow solid. MS: m / z = 162.1 (M+1, ESI+).

[0455] Preparation of intermediate B28: 7-nitro-1H-indole-2-carboxaldehyde

[0456]

[0457] Using the same method as that used to prepare intermediate B25, 7-nitro-1H-indole (4.4 g, 27.14 mmol) was used instead of 4,5,6,7-tetrahydro-1H-indole (2 g, 16.50 mmol) to give intermediate B28 (5 g, 96.90% yield) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 12.71 (s, 1H), 10.07 (s, 1H), 8.57 (d,1H), 8.48 (s, 1H), 8.24 (d, 1H), 7.47 (t, 1H); MS: m / z = 191.1 (M+1, ESI+).

[0458] Preparation of intermediate B29: 6-nitro-1H-indole-2-carboxaldehyde

[0459]

[0460] Using the same method as that used to prepare intermediate B25, 6-nitro-1H-indole (4.2 g, 25.90 mmol) was used instead of 4,5,6,7-tetrahydro-1H-indole (2 g, 16.50 mmol) to give intermediate B29 (4 g, 50.76% yield) as a yellow solid. MS: m / z = 191.1 (M+1, ESI+).

[0461] Preparation of intermediate B30: 4,5,6,7-tetrahydro-1H-benzo[d]imidazol-2-carboxaldehyde

[0462]

[0463] ​ ​

[0464] 4,5,6,7-Tetrahydro-1H-benzo[d]imidazole (2.2 g, 18.01 mmol) was dissolved in THF (20 mL), and then NaH (792 mg, 19.81 mmol, 60% purity) was added in portions at 0 °C, and the mixture was stirred at the same temperature for 1 hour. Subsequently, SEMCl (3.82 mL, 21.61 mmol) was added to the reaction mixture, and the mixture was stirred at room temperature for 16 hours. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (60 mL). The organic layer was washed three times with brine (300 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give 1-((2-(trimethylsilyl)ethoxy)methyl)-4,5,6,7-tetrahydro-1H-benzo[d]imidazole (1.5 g, 33.0% yield) as a yellow oil. MS: m / z = 253.3 (M+1, ESI+).

[0465] Step 2: Preparation of 1-((2-(trimethylsilyl)ethoxy)methyl)-4,5,6,7-tetrahydro-1H- benzo[d]imidazole-2-carbaldehyde Preparation of 2-formaldehyde

[0466] The same method used in step 2 for preparing intermediate B20 was employed to obtain 1-((2-(trimethylsilyl)ethoxy)methyl)-4,5,6,7-tetrahydro-1H-benzo[d]imidazol-2-carboxaldehyde (1.4 g, 84.01% yield) as a yellow oil, except that the product obtained in step 1 (1.5 g, 5.94 mmol) was used. MS: m / z = 281.2 (M+1, ESI+).

[0467] Step 3: Preparation of 4,5,6,7-tetrahydro-1H-benzo[d]imidazole-2-carbaldehyde

[0468] The product obtained in step 2 (1.4 g, 4.99 mmol) was dissolved in DCM (12 mL), and then TFA (4 mL) was added and stirred at 25 °C for 16 hours. Sodium bicarbonate solution (100 mL) was added to the reaction mixture, and the product was extracted three times with DCM (30 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, and concentrated to give intermediate B30 (700 mg, 93.37% yield) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 13.11 (s,1H), 9.46 (s, 1H), 2.56 (s, 4H), 1.76 (s, 4H); MS: m / z = 151.1 (M+1, ESI+).

[0469] Example 1: Preparation of N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-3-ethynyl-6-morpholinylimidazo[1,2-b]pyridazine-8-amine

[0470]

[0471] Step 1: Preparation of (6-morpholinylimidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester

[0472] Intermediate S1 (4.9 g, 11.57 mmol) was dissolved in methanol (70 mL), then Pd / C (1 g) and Pd(OH)2 / C (1 g) were added, and the mixture was stirred at 50 °C for 48 hours under a H2 atmosphere. After filtration and concentration of the reaction mixture, it was purified by silica gel chromatography to give (6-morpholinylimidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (3 g, yield 77.78%). MS: m / z = 334.1 (M+1, ESI+).

[0473] Step 2: Preparation of N-(tert-butoxycarbonyl)-N-(6-morpholinylimidazo[1,2-b]pyridazin-8- yl)glycine tert-butyl ester

[0474] The product obtained in step 1 (2.9 g, 10.26 mmol) and di-tert-butyl dicarbonate (8.95 g, 41.03 mmol) were dissolved in MeCN (60 mL), followed by the addition of cesium carbonate (13.37 g, 41.03 mmol) and stirring at 70 °C for 16 hours. Water (400 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (100 mL). The organic layer was washed three times with brine (400 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(tert-butyloxycarbonyl)-N-(6-morpholinylimidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (3.15 g, 80.22% yield). MS: m / z = 434.2 (M+1, ESI+).

[0475] Step 3: Preparation of N-(tert-butoxycarbonyl)-N-(3-iodo-6-morpholinylimidazo[1,2-b]pyridazin- 8-yl)glycine tert-butyl ester

[0476] The product obtained in step 2 (3.15 g, 8.23 ​​mmol) was dissolved in THF (50 mL), and then NIS (1.63 g, 7.27 mmol) was added at 0 °C and the mixture was stirred at 25 °C for 16 hours. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with sodium sulfite solution (400 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give N-(tert-butyloxycarbonyl)-N-(3-iodo-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (3.83 g, yield 94.22%). MS: m / z = 560.1 (M+1, ESI+).

[0477] Step 4: Preparation of N-(tert-butoxycarbonyl)-N-(6-moφholinyl-3-((trimethylsilyl)ethynyl)imidazo[1, 2-b]pyridazin-8-yl)glycine tert-butyl esters

[0478] The product obtained in step 3 (3.83 g, 6.85 mmol) and ethynyl(trimethyl)silane (1.01 g, 10.27 mmol) were dissolved in 1,4-dioxane (50 mL), followed by the addition of CuI (391 mg, 2.05 mmol), Pd(PPh3)2Cl2 (481 mg, 685 μmol), and TEA (2.86 mL, 20.54 mmol), and the mixture was stirred at 50 °C for 4 hours under a nitrogen atmosphere. Water (400 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (400 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(tert-butyloxycarbonyl)-N-(6-morpholinyl-3-((trimethylsilyl)ethynyl)imidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (2.86 g, yield 78.86%). MS: m / z = 530.2 (M+1, ESI+).

[0479] Step 5: Preparation of N-(tert-butoxycarbonyl)-N-(3-ethynyl-6-moφholinylimidazo[1,2-b]pyridazin- 8-yl)glycin tert-butyl ester

[0480] The product obtained in step 4 (2.86 g, 5.40 mmol) was dissolved in THF (30 mL), and then 1 MTBAF solution (21.6 mL, 21.60 mmol) was added and the mixture was stirred at 25 °C for 16 hours. An aqueous solution of ammonium chloride (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (60 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(tert-butyloxycarbonyl)-N-(3-ethynyl-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (2 g, yield 80.96%). MS: m / z = 458.2 (M+1, ESI+).

[0481] Step 6: Preparation of (3-ethynyl-6-moφholinylimidazo[1,2- b]pyridazin-8-yl)glycine

[0482] The product obtained in step 5 (2 g, 4.37 mmol) was dissolved in DCM (20 mL), and then TFA (3.34 mL, 43.71 mmol) was added. The mixture was stirred at 25 °C for 16 hours. After concentrating the reaction mixture, the concentrate was purified by silica gel chromatography to give (3-ethynyl-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)glycine (900 mg, yield 68.33%). MS: m / z = 302.1 (M+1, ESI+).

[0483] Step 7: Preparation of N-(6-amino-2,3-difluorophenyl)-2-((3-ethynyl-6-moφholinylimidazo[1,2- b]pyridazin-8-yl)amino)acetamide

[0484] The product obtained in step 6 (900 mg, 2.99 mmol) and intermediate B1 (517 mg, 3.58 mmol) were dissolved in DMF (20 mL), and then HATU (1.70 g, 4.48 mmol) and DIEA (1.56 mL, 8.96 mmol) were added. The mixture was stirred at 25 °C for 16 hours. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(6-amino-2,3-difluorophenyl)-2-((3-ethynyl-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)amino)acetamide (1 g, yield 78.33%). MS: m / z = 428.1 (M+1, ESI+).

[0485] Step 8: Preparation of N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-3-ethynyl-6-moφholinylimidazo [1,2-b]pyridazin-8-amine

[0486] The product obtained in step 7 (750 mg, 1.75 mmol) was dissolved in AcOH (10 mL), and the mixture was stirred at 70 °C for 16 hours. Sodium bicarbonate solution (80 mL) was added to the reaction mixture, and the product was extracted three times with DCM (30 mL). The organic layer was washed three times with brine (80 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by preparative HPLC to give the compound of Example 1 as a white solid (28 mg, yield 3.90%). 1 H NMR(400 MHz, DMSO-d6) δ 12.69 (s, 1H), 7.71-7.67 (m, 2H), 7.23-7.18 (m, 2H), 6.09 (s, 1H), 4.82 (d, 2H), 4.72 (s, 1H), 3.67 (s, 4H), 3.32 (s, 4H); MS: m / z= 410.0 (M+1, ESI+).

[0487] Example 2: Preparation of N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-6-morpholino-3-(2,2,2-trifluoroethyl)imidazo[1,2-b]pyridazine-8-amine

[0488]

[0489] Step 1: Preparation of 2-bromo-4,4,4-trifluorobutyraldehyde

[0490] 4,4,4-Trifluorobutanal (5 g, 39.66 mmol) was dissolved in 1,4-dioxane (50 mL), and then bromine (2.62 mL, 47.59 mmol) was added at 0 °C, and the mixture was stirred at 25 °C for 1 hour under a N2 atmosphere. Water (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (50 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, and concentrated to give 2-bromo-4,4,4-trifluorobutanal (4.5 g, crude product).

[0491] Step 2: Preparation of 8-bromo-6-chloro-3-(2,2,2-trifluoroethyl)imidazo[1,2-b]pyridazine

[0492] The product obtained in step 1 (4.5 g, 21.95 mmol) was dissolved in ethanol (50 mL), and then 4-bromo-6-chloro-pyridazin-3-amine (915 mg, 4.39 mmol) was added, and the mixture was stirred at 120 °C for 16 hours. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (100 mL). The organic layer was washed three times with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give 8-bromo-6-chloro-3-(2,2,2-trifluoroethyl)imidazo[1,2-b]pyridazin (890 mg, yield 64.45%). MS: m / z = 314.2 (M+1, ESI+).

[0493] Step 3: Preparation of (6-chloro-3-(2,2,2-trifluoroethyl)imidazol[1,2-b]pyridazin-8-yl)glycine tert butyl ester Step 4: Preparation of N-(6-chloro-3-(2,2,2-trifluoroethyl)imidazole[1,2-b]pyridazin-8-yl)-N-(4- methoxybenzyl)glycine tert-butyl ester

[0494] The product obtained in step 2 (890 mg, 2.83 mmol) was dissolved in glycine tert-butyl ester (10 mL) and stirred at 120 °C for 16 hours. Water (100 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (40 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give (6-chloro-3-(2,2,2-trifluoroethyl)imidazo[1,2- b 1,8-pyridazine-8-yl)glycine tert-butyl ester (770 mg, yield 74.60%). MS: m / z = 365.1 (M+1, ESI+).

[0495] Step 5: Preparation of N-(4-methoxybenzyl)-N-(6-moφholinyl-3-(2,2,2-trifluoroethyl)imidazo[1, 2-b]pyridazin-8-yl)glycine tert-butyl estere Step 6: Preparation of N-(4-methoxybenzyl)-N-(6-moφhoinyl-3-(2,2,2-trifluoroethyl)imidazo[1,

[0496] The product obtained in step 3 (770 mg, 2.11 mmol) and 4-methoxybenzyl chloride (496 mg, 3.17 mmol) were dissolved in MeCN (20 mL), and cesium carbonate (2.06 g, 6.33 mmol) was added. The mixture was stirred at 60 °C for 16 hours. Water (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (40 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(6-chloro-3-(2,2,2-trifluoroethyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine tert-butyl ester (470 mg, yield 45.91%). MS: m / z = 485.1 (M+1, ESI+).

[0497] 2-b]pyridazin-8-yl)glycin Step 7: Preparation of N-(6-amino-2,3-difluoroρhenyl)-2-((4-methoxybenzyl)(6-moφholinyl-3-(2,2,2- trifluoroethyl)imidazo[1,2-b]pyridazin-8-yl)amino)acetamide

[0498] The product obtained in step 4 (470 mg, 969 μmol) and morpholine (0.85 mL, 9.69 mmol) were dissolved in 1,4-dioxane (20 mL), followed by the addition of cesium carbonate (948 mg, 2.91 mmol) and RuPhos-Pd-G3 (81 mg, 97 μmol), and the mixture was stirred at 110 °C for 16 hours. Water (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (60 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give N-(4-methoxybenzyl)-N-(6-morpholinyl-3-(2,2,2-trifluoroethyl)imidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (290 mg, yield 55.87%). MS: m / z = 536.2(M+1, ESI+).

[0499] Step 8 : Preparation of N-((6,7-difluoro-1H-benzo[d ]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-6- moφholinyl-3-(2,2,2-trifluoroethyl)imidazol[1,2-b]pyridazin-8-amine ​

[0500] The product obtained in step 5 (290 mg, 541.5 μmol) was dissolved in 1,4-dioxane (5 mL) and water (2 mL), and then sodium hydroxide (109 mg, 2.71 mmol) was added. The mixture was stirred at 100 °C for 48 hours. 1 N hydrochloric acid (50 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (15 mL). The organic layer was washed three times with brine (50 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(4-methoxybenzyl)-N-(6-morpholino-3-(2,2,2-trifluoroethyl)imidazo[1,2-b]pyridazin-8-yl)glycine (220 mg, yield 84.74%). MS: m / z = 480.1 (M+1, ESI+).

[0501] ​ ​

[0502] The product obtained in step 6 (220 mg, 459 μmol) and intermediate B1 (2.20 g, 15.26 mmol) were dissolved in DMF (10 mL), and then HOBT (310 mg, 2.29 mmol), EDCI (440 mg, 2.29 mmol), and DIEA (0.4 mL, 2.29 mmol) were added. The mixture was stirred at 25 °C for 16 hours. Water (100 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (40 mL). The organic layer was washed three times with brine (100 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(6-amino-2,3-difluorophenyl)-2-((4-methoxybenzyl)(6-morpholino-3-(2,2,2-trifluoroethyl)imidazo[1,2-b]pyridazin-8-yl)amino)acetamide (230 mg, yield 82.77%). MS: m / z = 606.2 (M+1, ESI+).

[0503] ​ ​

[0504] The product obtained in step 7 (230 mg, 380 μmol) was dissolved in AcOH (10 mL), and the mixture was stirred at 70 °C for 16 hours. Sodium bicarbonate solution (60 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (20 mL). The organic layer was washed three times with brine (60 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-6-morpholino-3-(2,2,2-trifluoroethyl)imidazo[1,2-b]pyridazin-8-amine (300 mg, crude). MS: m / z = 588.2 (M+1, ESI+).

[0505] Step 9: Preparation of N-((6,7-difluoro-lH-benzo[d]imidazol-2-yl)methyl)-6- morpholino-3-(2,2,2-trifluoroethyl)imidazo[l,2-b]pyridazin-8-amine Step 1: Preparation of 2-bromo-3-methylbutanal

[0506] The product obtained in step 8 (300 mg, 509 μmol) was dissolved in DCM (10 mL), then TFA (5 mL) was added and the mixture was stirred at 25 °C for 16 hours. Sodium bicarbonate solution (100 mL) was added to the reaction mixture, and the product was extracted three times with DCM (40 mL). The organic layer was washed three times with brine (100 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by preparative HPLC to give the compound of Example 2 as a white solid (80 mg, yield 33.49%). 1 HNMR (400 MHz, DMSO-d6) δ 12.68 (s, 1H), 7.65 (t, 1H), 7.39 (s, 1H), 7.26-7.16(m, 2H), 6.04 (s, 1H), 4.82 (d, 2H), 3.95 (dd, 2H), 3.67 (t, 4H), 3.32 (t, 4H); MS: m / z = 468.1 (M+1, ESI+).

[0507] Example 3: Preparation of N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-3-isopropyl-6-morpholinylimidazo[1,2-b]pyridazine-8-amine

[0508]

[0509] Step 2: Preparation of 8-bromo-6-chloro-3-isopropylimidazo[l,2-b]pyridazine

[0510] 2-Bromo-methylbutyraldehyde (14.3 g, crude) was obtained by the same method as in step 1 of Example 2, except that 3-methylbutyraldehyde (11 g, 127.71 mmol) was dissolved in DCM (300 mL), and L-proline (1.47 g, 12.77 mmol) and NBS (29.55 g, 166.02 mmol) were used instead of bromine (7.61 g, 47.59 mmol), and the mixture was stirred for 16 hours.

[0511] Step 3 to 9: Preparation of N-((6,7-difluoro-lH-benzzo[d]imidazol-2-yl)methyl)-3- isopropyl-6-morpholinoimidazo[l,2-b]pyridazin-8-amine

[0512] The product obtained in step 1 (14.3 g, 86.65 mmol) was used to give 8-bromo-6-chloro-3-isopropylimidazo[1,2-b]pyridazine (4.3 g, 90.37% yield) in the same manner as in step 2 of Example 2, except that the reaction temperature was changed to 100 °C and the mixture was stirred for 72 hours. MS: m / z = 274.2 (M+1, ESI+).

[0513] Step 1 : Preparation of N-benzyl-N-(3-methyl-6-morpholinoimidazo[l,2-b]pyridazin-8- yl)glycine tert-butyl ester

[0514] Using the product obtained in step 2 as a starting material, the compound of Example 3 (210 mg, yield 67.26%) was obtained as a white solid by the same method as steps 3 to 9 of Example 2. However, the reaction times of steps 6, 7 and 9 of Example 2 were changed to 16 hours, 2 hours and 1 hour, respectively. 1 H NMR (400 MHz, DMSO-d6) δ 12.66(s, 1H), 7.50 (t, 1H), 7.22-7.13 (m, 3H), 5.97 (s, 1H), 4.79 (d, 2H), 3.68(t, 4H), 3.29 (t, 4H), 3.25-3.19 (m, 1H), 1.31 (d, 6H); MS: m / z = 428.2 (M+1,ESI+).

[0515] Example 4: Preparation of N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-3-methyl-6-morpholinylimidazo[1,2-b]pyridazine-8-amine

[0516]

[0517] Step 2: Preparation of (3-methyl-6-morpholinoimidazo[l,2-b]pyridazm-8-yl)glycine tert- butyl ester Step 3: Preparation of (3-methyl-6-morpholinoimidazo[l,2-b]-pyridazin-8-yl)glycine

[0518] Intermediate S2 (2.1 g, 4.18 mmol) and 2,4,6-trimethyl-1,3,5,2,4,6-trioxaborane (1.05 g, 8.36 mmol) were dissolved in 1,4-dioxane (20 mL) and water (5 mL), then Pd(PPh3)4 (483 mg, 418 μmol) and potassium carbonate (1.73 g, 12.54 mmol) were added, and the mixture was stirred at 100 °C for 16 hours. Water (100 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (30 mL). The organic layer was washed three times with brine (30 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-benzyl-N-(3-methyl-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (4.1 g, crude). MS: m / z = 438.2 (M+1, ESI+).

[0519] Step 4: Preparation of N-(6-amino-2,3-difluorophenyl)-2-((3-methyl-6-morpholinoimidazo[l,2- b]pyridazin-8-yl)amino)acetamide

[0520] The product obtained in step 1 (1.15 g, 2.63 mmol) was dissolved in methanol (20 mL), and then Pd / C (200 mg) and Pd(OH)2 / C (200 mg) were added and stirred at 50 °C for 48 hours under H2 atmosphere. The reaction mixture was filtered and concentrated, and then purified by silica gel chromatography to give (3-methyl-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (550 mg, yield 60.23%). MS: m / z = 348.2 (M+1, ESI+).

[0521] Step 5: Preparation of N-((6,7-difluoro-lH-benzo[d]-imidazol-2-yl)methyl)-3-methyl-6- morpholinoimidazo[l,2-b]pyridazin-8-amine

[0522] The product obtained in step 2 (550 mg, 1.58 mmol) was dissolved in DCM (10 mL), and then TFA (1.81 g, 15.83 mmol) was added and the mixture was stirred at 25 °C for 16 hours. The reaction mixture was concentrated to give (3-methyl-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)glycine (830 mg, crude). MS: m / z = 292.1 (M+1, ESI+).

[0523] Step l : Preparation of (6-morpholino-3-(trifluoromethyl)imidazo[l,2-b]pyridazin-8-yl)glycine tert- butyl ester Step 2: Preparation of (6-morpholino-3-(trifluoromethyl)imidazol[l,2-b]pyridazin-8-yl)glycine

[0524] The product obtained in step 3 (830 mg, 2.85 mmol) and intermediate B1 (493 mg, 3.42 mmol) were dissolved in anhydrous DMF (10 mL), and then HATU (1.63 g, 4.27 mmol) and DIEA (1.48 mL, 8.55 mmol) were added and the mixture was stirred at 25 °C for 16 hours. Water (100 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (30 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(6-amino-2,3-difluorophenyl)-2-((3-methyl-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)amino)acetamide (195 mg, yield 16.40%). MS: m / z = 418.1 (M+1, ESI+).

[0525] Step 3: Preparation of N-(6-amino-2,3-difluorophenyl) -2-((6-morpholino-3- (trifluoromethyl)imidazo[l,2-b]pyridazin- 8-yl)amino)acetamide Step 4: Preparation of N-((6,7-difluoro-lH-benzo[d])imidazol-2-yl)methyl)-6-morpholino-3- (trifluoromethyl)imidazo[l,2-b]pyridazin-8-amine

[0526] The product obtained in step 4 (195 mg, 467 μmol) was dissolved in AcOH (5 mL), and the mixture was stirred at 70 °C for 4 hours. Sodium bicarbonate solution (50 mL) was added to the reaction mixture, and the product was extracted three times with DCM (20 mL). The organic layer was washed three times with brine (50 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by preparative HPLC to give the compound of Example 4 as a white solid (70 mg, yield 37.52%). 1 H NMR (400MHz, DMSO-d6) δ 12.72 (s, 1H), 7.47 (s, 1H), 7.24-7.16 (m, 3H), 5.97 (s, 1H), 4.81 (d, 2H), 3.68 (t, 4H), 3.31 (t, 4H), 2.33 (s, 3H); MS: m / z = 400.1 (M+1,ESI+).

[0527] Example 5: Preparation of N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-3-ethyl-6-morpholinylimidazo[1,2-b]pyridazine-8-amine

[0528]

[0529] The compound of Example 5 (35 mg, yield 25.72%) was obtained as a white solid by the same method as steps 1 to 5 of Example 4, except that intermediate S2 (1.55 g, 3.09 mmol) was used as the starting material and vinylboric acid (266 mg, 3.70 mmol) was used instead of 2,4,6-trimethyl-1,3,5,2,4,6-trioxaborane (1.05 g, 8.36 mmol) in step 1 of Example 4. 1 H NMR (400 MHz, DMSO-d6) δ 12.66 (s, 1H), 7.48 (t,1H), 7.24-7.15 (m, 3H), 5.96 (s, 1H), 4.80 (d, 2H), 3.67 (t, 4H), 3.31 (t,4H), 2.78 (dd, 2H), 1.26 (t, 3H); MS: m / z = 414.0 (M+1, ESI+).

[0530] Example 6: Preparation of N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0531]

[0532] Step 1 : Preparation of N-(6-(3-oxa-8-azabicyclo[3.2. l]octan-8-yl)imidazo[l,2-b]pyridazin-8- yl)-N-benzyl glycine tert-butyl ester

[0533] Intermediate S3 (1.4 g, 2.85 mmol) was dissolved in methanol (20 mL), and then Pd / C (200 mg) and Pd(OH)2 / C (200 mg) were added. The mixture was stirred at 50 °C for 48 hours. The reaction mixture was filtered and concentrated, and then purified by silica gel chromatography to give (6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (910 mg, yield 79.59%). MS: m / z = 402.1 (M+1, ESI+).

[0534] Step 2: Preparation of N-(6-(3-oxa-8-azabicyclo[3,2,l]octan-8-yl)-3-iodoimidazo[l,2-b]pyridazin- 8-yl)-N-benzyl glycine tert-butyl ester

[0535] The product obtained in step 1 (910 mg, 2.27 mmol) was dissolved in DCM (10 mL), and then TFA (1.74 mL, 22.67 mmol) was added. The mixture was stirred at 25 °C for 16 hours. The reaction mixture was concentrated, and the concentrate was purified by silica gel chromatography to give (6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycine (970 mg, crude). MS: m / z = 346.1 (M+1, ESI+).

[0536] Step 3: Preparation of N-(6-(3-oxa-8-azabicyclo[ 3.2. l]octan-8-yl)-3-(trifluoromethyl)imidazo[l,2- b]pyridazin-8-yl)-N-benzyl glycine tert-butyl ester Step 4 to 7: Preparation of 6-(3-oxa-8-azabicyclo[3.2. l]-octan-8-yl)-N-((6,7-difluoro-lH-benzo[d] imidazol-2-yl)methyl)-3-(trifluoromethyl)imidazo[l,2-b]pyridazm-8-amine

[0537] The product obtained in step 2 (970 mg, 2.81 mmol) and intermediate B1 (486 mg, 3.37 mmol) were dissolved in anhydrous DMF (20 mL), and then HATU (1.60 g, 4.21 mmol) and DIEA (1.47 mL, 8.43 mmol) were added. The mixture was stirred at 25 °C for 16 hours. Water (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (40 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(6-amino-2,3-difluorophenyl)-2-((6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)amino)acetamide (890 mg, yield 67.21%). MS: m / z = 472.1 (M+1, ESI+).

[0538] ​ ​

[0539] The product obtained in step 3 (890 mg, 1.89 mmol) was dissolved in AcOH (10 mL), and the mixture was stirred at 70 °C for 4 hours. Sodium bicarbonate solution (80 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (30 mL). The organic layer was washed three times with brine (100 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by preparative HPLC to give the compound of Example 6 as a white solid (570 mg, yield 66.59%). 1H NMR(400 MHz, DMSO-d6) δ 12.83 (s, 1H), 7.88-7.84 (m, 2H), 7.28-7.15 (m, 2H), 6.21 (s, 1H), 4.84 (d, 2H), 3.67 (t, 4H), 3.35 (t, 4H); MS: m / z = 454.0 (M+1,ESI+).

[0540] Example 7: Preparation of 6-(3-oxa-8-azabicyclo[3.2.1]octane-8-yl)-N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine

[0541]

[0542] ​ ​

[0543] The product obtained in step 2 of intermediate S1 (6 g, 16 mmol) and intermediate A1 (2.73 g, 24 mmol) were dissolved in 1,4-dioxane (60 mL), and then RuPhos-Pd-G3 (674 mg, 805 μmol) and cesium carbonate (15.73 g, 48.28 mmol) were added. The mixture was stirred at 110 °C for 16 hours under a nitrogen atmosphere. Water (400 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (400 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(6-(3-oxa-8-azabicyclo[3.2.1]octane-8-yl)imidazo[1,2-b]pyridazin-8-yl)-N-benzylglycine tert-butyl ester (5.1 g, yield 70.50%). MS: m / z = 450.2 (M+1, ESI+).

[0544] ​ ​

[0545] The product obtained in step 1 (5.1 g, 11.34 mmol) was dissolved in THF (30 mL), and then NIS (3.06 g, 13.61 mmol) was added at 0 °C and stirred at 25 °C for 16 hours. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with DCM (80 mL). The organic layer was washed three times with brine (400 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give N-(6-(3-oxa-8-azabicyclo[3.2.1]octane-8-yl)-3-iodomizo[1,2-b]pyridazin-8-yl)-N-benzylglycine tert-butyl ester (5.2 g, yield 79.65%). MS: m / z = 576.1 (M+1, ESI+).

[0546] ​ ​

[0547] The product obtained in step 2 (5.2 g, 9.04 mmol) and methyl 2,2-difluoro-2-(fluorosulfonyl)acetate (17.36 g, 90.4 mmol) were dissolved in DMF (30 mL), and then CuI (8.61 g, 45.2 mmol) was added and the mixture was stirred at 100 °C for 16 hours. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (300 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give N-(6-(3-oxa-8-azabicyclo[3.2.1]octane-8-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)-N-benzylglycine tert-butyl ester (2.8 g, yield 59.87%). MS: m / z = 518.5 (M+1, ESI+).

[0548] ​ ​

[0549] The product of step 3 was used as a starting material to obtain the compound of Example 7 (58 mg, yield 14.33%) as a white solid using the same method as steps 1 to 4 of Example 6. However, the reaction time of step 7 of Example 7 was changed to 2 hours. 1H NMR (400 MHz, DMSO-d6) δ 12.75 (s, 1H), 7.89-7.87 (m, 2H), 7.28-7.16(m, 2H), 6.13 (s, 1H), 4.84 (d, 2H), 4.28 (s, 2H), 3.61 (d, 2H), 3.44 (d,2H), 1.94-1.83 (m, 4H); MS: m / z = 480.1 (M+1, ESI+).

[0550] In the following text, the compounds of Examples 8 to 22 were prepared using appropriate intermediates from the intermediate series A in [Table 1] according to Scheme I.

[0551] [Option I]

[0552]

[0553] In scheme I above, R 2 Having R with the compounds of each embodiment 2 The same structure. The intermediates referred to as intermediates A1 to A52 described in the following [Table 1] have the structure shown in the following [Table 1], throughout the embodiments and embodiments 8 to 22.

[0554] [Table 1]

[0555]

[0556]

[0557] Example 8: Preparation of N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine

[0558]

[0559] Step 1: N-(4-methoxybenzyl)-N-(6-(4-methylpiperazin-1-yl)imidazo[1,2-b]pyridazin-8-yl) Preparation of glycine tert-butyl ester

[0560] Intermediate S4 (6.1 g, 15.14 mmol) and intermediate A2 (4.54 g, 45.42 mmol) were dissolved in 1,4-dioxane (100 mL), followed by the addition of cesium carbonate (9.87 g, 30.28 mmol) and Ruphos-Pd-G3 (1.27 g, 1.51 mmol). The mixture was stirred at 100 °C for 16 hours under a nitrogen atmosphere. Water (500 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (100 mL). The organic layer was washed three times with brine (500 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(4-methoxybenzyl)-N-(6-(4-methylpiperazin-1-yl)imidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (3.9 g, yield 55.21%). MS: m / z = 467.3 (M+1, ESI+).

[0561] Step 2: N-(3-iodo-6-(4-methylpiperazin-1-yl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxy) Preparation of benzyl glycine tert-butyl ester

[0562] The product obtained in step 1 (3.8 g, 8.14 mmol) was dissolved in DCM (80 mL), and then NIS (1.03 g, 4.59 mmol) and TFA (1.25 mL, 16.28 mmol) were added to it at 0 °C. The mixture was stirred at 25 °C for 2 hours under a N2 atmosphere. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with DCM (60 mL). The organic layer was washed three times again with sodium sulfite solution (300 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(3-iodo-6-(4-methylpiperazin-1-yl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine tert-butyl ester (2.3 g, yield 47.66%). MS: m / z = 593.2 (M+1, ESI+).

[0563] Step 3: N-(4-methoxybenzyl)-N-(6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-] Preparation of [b]pyridazine-8-yl)glycine tert-butyl ester

[0564] The product obtained in step 2 (2.1 g, 3.85 mmol) and methyl 2,2-difluoro-2-(fluorosulfonyl)acetate (7.40 g, 38.50 mmol) were dissolved in N-methyl-2-pyrrolidone (35 mL), and then CuI (3.67 g, 19.25 mmol) and HMPA (3.45 g, 19.25 mmol) were added. The mixture was stirred at 50 °C for 16 hours. Water (400 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (400 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(4-methoxybenzyl)-N-(6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (850 mg, yield 41.30%). MS: m / z = 535.3 (M+1, ESI+).

[0565] Step 4: N-(4-methoxybenzyl)-N-(6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-] Preparation of [b]pyridazine-8-yl)glycine

[0566] The product obtained in step 3 (850 mg, 1.59 mmol) was dissolved in 1,4-dioxane (10 mL) and water (10 mL), and then sodium hydroxide (1.27 g, 31.80 mmol) was added. The mixture was stirred at 100 °C for 16 hours. 0.5 N hydrochloric acid (80 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (30 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(4-methoxybenzyl)-N-(6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycine (500 mg, yield 65.72%). MS: m / z = 479.2 (M+1, ESI+).

[0567] Step 5: N-(6-amino-2,3-difluorophenyl)-2-((4-methoxybenzyl)(6-(4-methylpiperazin-1-yl)- Preparation of 3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)amino)acetamide

[0568] The product obtained in step 4 (500 mg, 1.05 mmol) and intermediate B1 (181 mg, 1.26 mmol) were dissolved in DMF (15 mL), and then HOBT (706 mg, 5.23 mmol), EDCI (1.00 g, 5.23 mmol), and DIEA (1.82 mL, 10.45 mmol) were added. The mixture was stirred at 25 °C for 2 hours. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(6-amino-2,3-difluorophenyl)-2-((4-methoxybenzyl)(6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)amino)acetamide (380 mg, yield 60.15%). MS: m / z = 605.2 (M+1, ESI+).

[0569] Step 6: N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-6-(4-methyl) Preparation of pyridazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine

[0570] The product obtained in step 5 (380 mg, 629 μmol) was dissolved in AcOH (10 mL), and the mixture was stirred at 100 °C for 2 hours. Sodium bicarbonate solution (100 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (40 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (330 mg, crude). MS: m / z = 587.2 (M+1, ESI+).

[0571] Step 7: N-((6) , 7-Difluoro-1H-benzo[d]imidazol-2-yl)methyl)-6-(4-methylpiperazin-1-yl)-3- Preparation of (trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0572] The product obtained in step 6 (330 mg, 562.60 μmol) was dissolved in DCM (10 mL), and then TFA (5 mL) was added. The mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated and then purified by preparative HPLC to give the compound of Example 8 (55 mg, yield 20.96%) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 12.71 (s, 1H), 7.86 (s, 1H), 7.82 (t, 1H), 7.27-7.15 (m, 2H), 6.19 (s,1H), 4.83 (d, 2H), 3.37 (t, 4H), 2.35 (t, 4H), 2.18 (s, 3H); MS: m / z = 467.2(M+1, ESI+).

[0573] Examples 9 to 18

[0574] The compounds of Examples 9 to 18 were prepared using the intermediates described in [Table 2] in the same manner as in Example 8.

[0575] [Table 2]

[0576]

[0577]

[0578]

[0579] Example 19: Preparation of 1-(8-(((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)piperidine-4-ol

[0580]

[0581] Step 1: N-(4-methoxybenzyl)-N-(6-(4-oxopiperidin-1-yl)imidazo[1,2-b]pyridazin-8-yl) Preparation of glycine tert-butyl ester

[0582] Using intermediates S4 (9 g, 22.34 mmol) and A13 (3.32 g, 33.51 mmol), N-(4-methoxybenzyl)-N-(6-(4-oxopiperidin-1-yl)imidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (5.4 g, yield 51.92%) was obtained in the same manner as in step 1 of Example 8, except that the reaction was carried out at 110 °C. MS: m / z = 466.2 (M+1, ESI+).

[0583] Step 2: N-(3-iodo-6-(4-oxopiperidin-1-yl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxy) Preparation of benzyl glycine tert-butyl ester

[0584] N-(3-iodo-6-(4-oxopiperidin-1-yl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine tert-butyl ester (2.8 g, yield 40.81%) was obtained by the same method as in step 2 of Example 8, except that THF (50 mL) was used instead of DCM (80 mL), TFA was not added, and the mixture was stirred at 0 °C for 1 hour. MS: m / z = 592.3 (M+3, ESI+).

[0585] Step 3: N-(4-methoxybenzyl)-N-(6-(4-oxopiperidin-1-yl)-3-(trifluoromethyl)imidazo[1,2- Preparation of [b]pyridazine-8-yl)glycine tert-butyl ester

[0586] N-(4-methoxybenzyl)-N-(6-(4-oxopiperidin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (1.7 g, yield 67.30%) was obtained by the same method as in step 3 of Example 8, except that DMF (80 mL) was used instead of NMP (35 mL) at 100 °C, and DIEA (11.3 mL, 64.67 mmol) was used instead of HMPA (3.45 g, 19.25 mmol). MS: m / z = 534.3 (M+1, ESI+).

[0587] Step 4: N-(4-methoxybenzyl)-N-(6-(4-oxopiperidin-1-yl)-3-(trifluoromethyl)imidazo[1,2-] Preparation of [b]pyridazine-8-yl)glycine

[0588] N-(4-methoxybenzyl)-N-(6-(4-oxopiperidin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycine (1.2 g, yield 78.88%) was obtained by the same method as in step 4 of Example 8, except that the reaction time was changed to 2 hours. MS: m / z = 492.2 (M+1, ESI+).

[0589] Step 5: N-(6-amino-2,3-difluorophenyl)-2-((4-methoxybenzyl)(6-(4-oxopiridin-1-yl)- Preparation of 3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)amino)acetamide

[0590] N-(6-amino-2,3-difluorophenyl)-2-((4-methoxybenzyl)(6-(4-oxopiperidin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)amino)acetamide (1.1 g, yield 72.85%) was obtained using the same method as in step 5 of Example 8, except that HATU (1.14 g, 3.01 mmol) was used instead of HOBT (706 mg, 5.23 mmol) and EDCI (1.00 g, 5.23 mmol). MS: m / z = 604.1 (M+1, ESI+).

[0591] Step 6: 1-(8-(((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)- Preparation of 3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)piperidin-4-one

[0592] 1-(8-(((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)piperidin-4-one (700 mg, yield 65.60%) was obtained by the same method as in step 6 of Example 8, except that the reaction temperature was changed to 70°C and the mixture was stirred for 16 hours. MS: m / z = 586.2 (M+1, ESI+).

[0593] Step 7: 1-(8-(((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)- Preparation of 3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)piperidine-4-ol

[0594] The product obtained in step 6 (700 mg, 1.2 mmol) was dissolved in methanol (10 mL), and sodium borohydride (55 mg, 1.44 mmol) was added at 0 °C and stirred at 25 °C for 2 hours. The reaction mixture was concentrated and the concentrate was purified by preparative HPLC to give 1-(8-(((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)piperidine-4-ol (550 mg, 78.57% yield). MS: m / z = 588.5 (M+1, ESI+).

[0595] Step 8: 1-(8-(((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazolium Preparation of azido[1,2-b]pyridazine-6-yl)piperidine-4-ol

[0596] The compound of Example 19 (200 mg, yield 45.76%) was obtained as a white solid by the same method as step 7 of Example 8, except that the reaction time was changed to 2 hours. 1 H NMR (400 MHz, DMSO-d6) δ 12.73(s, 1H), 7.85-7.80 (m, 2H), 7.25-7.16 (m, 2H), 6.16 (s, 1H), 4.83 (d, 2H),4.67 (d, 1H), 3.80 (d, 2H), 3.66-3.63 (m, 1H), 3.02 (t, 2H), 1.71 (t, 2H), 1.34-1.27 (m, 2H); MS: m / z = 468.3 (M+1, ESI+).

[0597] Example 20: Preparation of 6-(2-aminopyrimidin-5-yl)-N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-3-(prop-1-yn-1-yl)imidazo[1,2-b]pyridazine-8-amine

[0598]

[0599] Step 1: N-(6-(2-aminopyrimidin-5-yl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl) Preparation of glycine tert-butyl ester

[0600] Intermediate S4 (2.6 g, 6.45 mmol) and intermediate A14 (1.34 g, 9.68 mmol) were dissolved in 1,4-dioxane (50 mL) and water (50 mL), then [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride (469 mg, 645 μmol) and cesium carbonate (4.21 g, 12.91 mmol) were added, and the mixture was stirred at 100 °C for 16 hours under a nitrogen atmosphere. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(6-(2-aminopyrimidin-5-yl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine tert-butyl ester (2.5 g, yield 83.94%). MS: m / z = 462.2 (M+1, ESI+).

[0601] Step 2: N-(6-(2-aminopyrimidin-5-yl)-3-bromoimidazolo[1,2-b]pyridazin-8-yl)-N-(4-methoxy) Preparation of benzyl glycine tert-butyl ester

[0602] The product obtained in step 1 (2.5 g, 5.42 mmol) was dissolved in THF (50 mL), and then NBS (964 mg, 5.42 mmol) was added. The mixture was stirred at -60 °C for 3 hours under a N2 atmosphere. Water (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (50 mL). The organic layer was washed three times again with sodium sulfite solution (200 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(6-(2-aminopyrimidin-5-yl)-3-bromoimidazolo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine tert-butyl ester (2.5 g, yield 85.40%). MS: m / z = 542.2 (M+3, ESI+).

[0603] Step 3: N-(6-(2-aminopyrimidin-5-yl)-3-(prop-1-yn-1-yl)imidazo[1,2-b]pyridazin-8-yl)- Preparation of N-(4-methoxybenzyl)glycine tert-butyl ester

[0604] The product obtained in step 2 (2.5 g, 4.63 mmol) and propyne (3.71 g, 92.52 mmol) were dissolved in DMF (20 mL), and then CuI (176 mg, 925 μmol), Pd(PPh3)2Cl2 (325 mg, 463 μmol) and TEA (0.65 mL, 4.63 mmol) were added. The mixture was stirred at 40 °C for 72 hours under a nitrogen atmosphere. Water (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (60 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to obtain N-(6-(2-aminopyrimidin-5-yl)-3-(prop-1-yn-1-yl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine. Uncle Butyl ester (1.1 g, yield 47.60%). MS: m / z = 500.2 (M+1, ESI+).

[0605] Step 4: N-(6-(2-aminopyrimidin-5-yl)-3-(prop-1-yn-1-yl)imidazo[1,2-b]pyridazin-8-yl)- Preparation of N-(4-methoxybenzyl)glycine

[0606] The product obtained in step 3 (1.1 g, 2.20 mmol) was used to give N-(6-(2-aminopyrimidin-5-yl)-3-(prop-1-yn-1-yl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine (300 mg, yield 30.72%) using the same method as in step 4 of Example 8. MS: m / z = 442.2 (M+1, ESI+).

[0607] Step 5: N-(6-amino-2,3-difluorophenyl)-2-((6-(2-aminopyrimidin-5-yl)-3-(prop-1-yn-1-yl) Preparation of (-)imidazo[1,2-b]pyridazin-8-yl)(4-methoxybenzyl)amino)acetamide

[0608] N-(6-amino-2,3-difluorophenyl)-2-((6-(2-aminopyrimidin-5-yl)-3-(prop-1-yn-1-yl)imidazo[1,2-b]pyridazin-8-yl)(4-methoxybenzyl)amino)acetamide (270 mg, yield 70.07%) was obtained by the same method as in step 5 of Example 8, except that DMF (15 mL) was replaced with DCM (20 mL) and the mixture was stirred for 16 hours. MS: m / z = 570.3 (M+1, ESI+).

[0609] Step 6: 6-(2-aminopyrimidin-5-yl)-N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-N- Preparation of (4-methoxybenzyl)-3-(prop-1-yn-1-yl)imidazo[1,2-b]pyridazine-8-amine

[0610] 6-(2-aminopyrimidin-5-yl)-N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-3-(prop-1-yn-1-yl)imidazo[1,2-b]pyridazin-8-amine (250 mg, 95.62% yield) was obtained by the same method as in step 6 of Example 8, except that the reaction temperature was changed to 70°C and the mixture was stirred for 16 hours. MS: m / z = 552.2 (M+1, ESI+).

[0611] Step 7: 6-(2-aminopyrimidin-5-yl)-N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-3- Preparation of (prop-1-yn-1-yl)imidazo[1,2-b]pyridazine-8-amine

[0612] The compound of Example 20 (8 mg, yield 4.09%) was obtained as a white solid by the same method as step 7 of Example 8. 1 H NMR (400 MHz, DMSO-d6) δ 12.73 (s, 1H), 8.79 (s, 2H), 8.07 (s,1H), 7.73 (s, 1H), 7.22-7.08 (m, 4H), 6.72 (s, 1H), 4.94 (d, 2H), 2.21 (s,3H); MS: m / z = 432.1 (M+1, ESI+).

[0613] Example 21: Preparation of 6-(2-aminopyrimidin-5-yl)-3-(cyclopropylethynyl)-N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)imidazo[1,2-b]pyridazin-8-amine

[0614]

[0615] Step 1: N-(6-(2-aminopyrimidin-5-yl)-3-(cyclopropylethynyl)imidazo[1,2-b]pyridazin-8-yl)- Preparation of N-(4-methoxybenzyl)glycine tert-butyl ester

[0616] The product obtained in step 2 of Example 20 (2.6 g, 4.81 mmol) and ethynylcyclopropane (318 mg, 4.81 mmol) were dissolved in MeCN (20 mL), and then tris(dibenzylacetone)dipalladium(O) (441 mg, 481 μmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (X-phos, 459 mg, 962 μmol) and potassium phosphate (K3PO4, 3.06 g, 14.43 mmol) were added. The mixture was stirred at 70 °C for 48 hours under a nitrogen atmosphere. Water (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (60 mL). The organic layer was washed three times with brine (200 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(6-(2-aminopyrimidin-5-yl)-3-(cyclopropylethynyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine tert-butyl ester (900 mg, yield 35.59%). MS: m / z = 526.3 (M+1, ESI+).

[0617] Step 2: N-(6-(2-aminopyrimidin-5-yl)-3-(cyclopropylethynyl)imidazo[1,2-b]pyridazin-8-yl)- Preparation of N-(4-methoxybenzyl)glycine

[0618] N-(6-(2-aminopyrimidin-5-yl)-3-(cyclopropylethynyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine (780 mg, yield 67.17%) was obtained using the same method as in step 4 of Example 8. MS: m / z = 470.2 (M+1, ESI+).

[0619] Step 3: N-(6-amino-2,3-difluorophenyl)-2-((6-(2-aminopyrimidin-5-yl)-3-(cyclopropylacetylene) Preparation of (-)imidazo[1,2-b]pyridazin-8-yl)(4-methoxybenzyl)amino)acetamide

[0620] N-(6-amino-2,3-difluorophenyl)-2-((6-(2-aminopyrimidin-5-yl)-3-(cyclopropylethynyl)imidazo[1,2-b]pyridazin-8-yl)(4-methoxybenzyl)amino)acetamide (660 mg, yield 66.70%) was obtained by the same method as in step 5 of Example 8, except that DCM (20 mL) was used instead of DMF (15 mL) and the mixture was stirred for 16 hours. MS: m / z = 596.2 (M+1, ESI+).

[0621] Step 4: 6-(2-aminopyrimidin-5-yl)-3-(cyclopropylethynyl)-N-((6,7-difluoro-1H-benzo[d]imidazolium) Preparation of (2-yl)-N-(4-methoxybenzyl)imidazo[1,2-b]pyridazine-8-amine

[0622] 6-(2-aminopyrimidin-5-yl)-3-(cyclopropylethynyl)-N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)imidazo[1,2-b]pyridazin-8-amine (580 mg, 90.62% yield) was obtained by the same method as in step 6 of Example 8, except that the reaction temperature was changed to 70°C and the mixture was stirred for 16 hours. MS: m / z = 578.2 (M+1, ESI+).

[0623] Step 5: 6-(2-aminopyrimidin-5-yl)-3-(cyclopropylethynyl)-N-((6,7-difluoro-1H-benzo[d]imidazolium) Preparation of (-2-yl)methyl)imidazo[1,2-b]pyridazin-8-amine

[0624] The compound of Example 21 (25 mg, yield 5.44%) was obtained as a white solid by the same method as step 7 of Example 8. 1 H NMR (400 MHz, DMSO-d6) δ 12.73 (s, 1H), 8.80 (s, 2H), 8.07-8.03(m, 1H), 7.72 (s, 1H), 7.24-7.17 (m, 2H), 7.08 (s, 2H), 6.73 (s, 1H), 4.93(s, 2H), 1.73-1.69 (m, 1H), 0.99-0.95 (m, 2H), 0.84-0.80 (m, 2H); MS: m / z =458.2 (M+1, ESI+).

[0625] Example 22: Preparation of 6-(2-aminopyrimidin-5-yl)-N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-3-ethynylimidazo[1,2-b]pyridazine-8-amine

[0626]

[0627] Step 1: N-(6-(2-aminopyrimidin-5-yl)-3-ioimidozolo[1,2-b]pyridazin-8-yl)-N-(4-methoxy) Preparation of benzyl glycine tert-butyl ester

[0628] N-(6-(2-aminopyrimidin-5-yl)-3-ioimidazolo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine tert-butyl ester (3.7 g, yield 72.68%) was obtained using the same method as in step 2 of Example 8, except that the product obtained in step 1 of Example 20 (4 g, 8.67 mmol) was used as the starting material, and the reaction time was changed to 1 hour. MS: m / z = 588.2 (M+1, ESI+).

[0629] Step 2: N-(6-(2-aminopyrimidin-5-yl)-3-((trimethylsilyl)ethynyl)imidazo[1,2-b] Preparation of tert-butyl pyridazine-8-yl)-N-(4-methoxybenzyl)glycine

[0630] The product obtained in step 1 (3.7 g, 6.30 mmol) and ethynyl(trimethyl)silane (928 mg, 9.45 mmol) were dissolved in DMF (50 mL), and then CuI (240 mg, 1.26 mmol), Pd(PPh3)2Cl2 (577 mg, 630 μmol) and TEA (2.63 mL, 18.90 mmol) were added. The mixture was stirred at 50 °C for 2 hours under a nitrogen atmosphere. Water (500 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (100 mL). The organic layer was washed three times with brine (500 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(6-(2-aminopyrimidin-5-yl)-3-((trimethylsilyl)ethynyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine tert-butyl ester (2.9 g, yield 82.55%). MS: m / z = 558.5 (M+1, ESI+).

[0631] Steps 3 to 6: 6-(2-aminopyrimidin-5-yl)-N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)- Preparation of 3-ethynylimidazo[1,2-b]pyridazine-8-amine

[0632] The compound of Example 22 (30 mg, yield 9.66%) was obtained as a white solid using the same method as steps 4 to 7 of Example 8. However, the reaction time in step 5 of Example 8 was changed to 16 hours, and the reaction temperature in step 6 of Example 8 was changed to 40°C and stirred for 16 hours. 1 H NMR (400 MHz, DMSO-d6) δ 12.74 (s, 1H),8.80 (s, 2H), 8.15 (s, 1H), 7.88 (s, 1H), 7.22-7.11 (m, 4H), 6.78 (s, 1H),4.95 (d, 2H), 4.86 (s, 1H); MS: m / z = 418.1 (M+1, ESI+).

[0633] Example 23: Preparation of 3-cyclopropyl-N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-6-morpholinylimidazo[1,2-b]pyridazine-8-amine

[0634]

[0635] Step 1: N-(3-cyclopropyl-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine Preparation of amino acid tert-butyl ester

[0636] Intermediate S5 (2.6 g, 4.88 mmol) and cyclopropylboronic acid (1.26 g, 14.65 mmol) were dissolved in 1,4-dioxane / water (40 mL / 10 mL), and then Pd(PPh3)4 (564 mg, 488 μmol) and potassium carbonate (K2CO3, 2.02 g, 14.65 mmol) were added. The mixture was stirred at 110 °C for 16 hours. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (300 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give N-(3-cyclopropyl-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine tert-butyl ester (1.47 g, yield 60.99%). MS: m / z = 494.2 (M+1, ESI+).

[0637] Step 2: N-(3-cyclopropyl-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine Preparation of amino acids

[0638] The product obtained in step 1 (1.47 g, 2.98 mmol) was dissolved in 1,4-dioxane / water (10 mL / 4 mL), and then N-(3-cyclopropyl-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine (620 mg, yield 47.59%) was obtained by the same method as in step 4 of Example 8. MS: m / z = 438.2 (M+1, ESI+).

[0639] Step 3: N-(6-amino-2,3-difluorophenyl)-2-((3-cyclopropyl-6-morpholinylimidazo[1,2-b]pyridazine- Preparation of 8-yl)(4-methoxybenzyl)amino)acetamide

[0640] The product obtained in step 2 (620 mg, 1.42 mmol) and intermediate B1 (245 mg, 1.70 mmol) were dissolved in DMF (15 mL), and then HATU (802 mg, 2.13 mmol) and DIEA (550 mg, 4.25 mmol) were added. The mixture was stirred at 70 °C for 16 hours. Water (150 mL) was added to the reaction mixture, and the product was extracted three times with DCM (40 mL). The organic layer was washed three times with brine (150 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(6-amino-2,3-difluorophenyl)-2-((3-cyclopropyl-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)(4-methoxybenzyl)amino)acetamide (410 mg, yield 51.33%). MS: m / z = 564.2 (M+1, ESI+).

[0641] Step 4: 3-Cyclopropyl-N-((6,7-Difluoro-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl) Preparation of 6-morpholinylimidazo[1,2-b]pyridazine-8-amine

[0642] 3-Cyclopropyl-N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-6-morpholinylimidazo[1,2-b]pyridazin-8-amine (350 mg, 87.88% yield) was obtained by the same method as in step 6 of Example 8, except that the reaction temperature was changed to 70°C and the mixture was stirred for 16 hours. MS: m / z = 546.2 (M+1, ESI+).

[0643] Step 5: 3-Cyclopropyl-N-((6,7-Difluoro-1H-benzo[d]imidazol-2-yl)methyl)-6-morpholinylimidazol Preparation of [1,2-b]pyridazine-8-amine

[0644] The product obtained in step 4 (350 mg, 642 μmol) was dissolved in TFA (5 mL) and then stirred at 25 °C for 1 hour. The reaction mixture was concentrated and the concentrate was purified by preparative HPLC to give the compound of Example 23 (120 mg, yield 43.97%) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 12.70 (s, 1H), 7.47 (s,1H), 7.26-7.16 (m, 2H), 7.09 (s, 1H), 6.00 (s, 1H), 4.80 (d, 2H), 3.68 (t,4H), 3.34 (t, 4H), 2.09-2.04 (m, 1H), 0.96-0.92 (m, 2H), 0.80-0.76 (m, 2H); MS: m / z = 426.2 (M+1, ESI+).

[0645] Example 24: Preparation of N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-6-morpholino-3-(prop-1-yn-1-yl)imidazo[1,2-b]pyridazine-8-amine

[0646]

[0647] Step 1: N-(4-methoxybenzyl)-N-(6-morpholino-3-(prop-1-yn-1-yl)imidazo[1,2-b]pyridazine- Preparation of 8-yl)glycine tert-butyl ester

[0648] Intermediate S5 (4 g, 7.51 mmol) and propyne (3 g, 75.1 mmol) were dissolved in DMF (30 mL), and then CuI (429 mg, 2.25 mmol), Pd(PPh3)2Cl2 (527 mg, 751 μmol) and TEA (3.14 mL, 22.54 mmol) were added. The mixture was stirred at 40 °C for 48 hours under a nitrogen atmosphere. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (80 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(4-methoxybenzyl)-N-(6-morpholino-3-(prop-1-yn-1-yl)imidazo[1,2-b]pyridazin-8-yl)glycine tert-butyl ester (1.45 g, yield 39.26%). MS: m / z = 492.2 (M+1, ESI+).

[0649] Step 2: N-(4-methoxybenzyl)-N-(6-morpholino-3-(prop-1-yn-1-yl)imidazo[1,2-b]pyridazine- Preparation of 8-yl)glycine

[0650] The product obtained in step 1 (1.45 g, 2.95 mmol) was dissolved in 1,4-dioxane / water (15 mL / 15 mL), and then N-(4-methoxybenzyl)-N-(6-morpholino-3-(prop-1-yn-1-yl)imidazo[1,2-b]pyridazin-8-yl)glycine (950 mg, yield 73.96%) was obtained by the same method as in step 4 of Example 8. MS: m / z = 436.2 (M+1, ESI+).

[0651] Step 3: N-(6-amino-2,3-difluorophenyl)-2-((4-methoxybenzyl)(6-morpholino-3-(prop-1-yne-) Preparation of 1-yl)imidazo[1,2-b]pyridazine-8-yl)amino)acetamide

[0652] The product obtained in step 2 (900 mg, 2.07 mmol) was dissolved in DMF (15 mL), and then 1,1-carbonyldiimidazole (312 mg, 2.17 mmol) was added. The mixture was stirred at 25 °C for 2 hours. Intermediate B1 (298 mg, 2.07 mmol) was added to the reaction solution, and the mixture was stirred at 50 °C for 16 hours. Water (150 mL) was added to the reaction mixture, and the product was extracted three times with DCM (40 mL). The organic layer was washed three times with brine (150 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give N-(6-amino-2,3-difluorophenyl)-2-((4-methoxybenzyl)(6-morpholino-3-(prop-1-yn-1-yl)imidazo[1,2-b]pyridazin-8-yl)amino)acetamide (470 mg, yield 40.50%). MS: m / z = 562.2 (M+1, ESI+).

[0653] Step 4: N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-6-(morpholine) Preparation of 3-(prop-1-yn-1-yl)imidazo[1,2-b]pyridazine-8-amine

[0654] N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-6-(morpholino-3-(prop-1-yn-1-yl)imidazo[1,2-b]pyridazin-8-amine (410 mg, 90.13% yield) was obtained by the same method as in step 6 of Example 8, except that the reaction mixture was stirred for 1 hour. MS: m / z = 520.2 (M+1, ESI+).

[0655] Step 5: N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-6-morpholino-3-(prop-1-yne-1-) Preparation of (1,2-b)imidazo[1,2-b]pyridazine-8-amine

[0656] The product obtained in step 4 (410 mg, 754 μmol) was dissolved in TFA (10 mL) and then stirred at 25 °C for 1 hour. The reaction mixture was concentrated and the concentrate was purified by preparative HPLC to give the compound of Example 24 (50 mg, yield 15.67%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.63 (t, 1H), 7.55 (s,1H), 7.28-7.19 (m, 2H), 6.06 (s, 1H), 4.81 (d, 2H), 3.68 (t, 4H), 3.39 (t,4H), 2.15 (s, 3H); MS: m / z = 424.2 (M+1, ESI+).

[0657] Example 25: Preparation of 3-(cyclopropylethynyl)-N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-6-morpholinylimidazo[1,2-b]pyridazine-8-amine

[0658]

[0659] Step 1: N-(3-(cyclopropylethynyl)-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxy) Preparation of benzyl glycine tert-butyl ester

[0660] N-(3-(cyclopropylethynyl)-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine tert-butyl ester (2.66 g, 65.15% yield) was obtained by the same method as in step 1 of Example 24, except that ethynylcyclopropane (782 mg, 11.83 mmol) was used instead of prop-1-yne (3 g, 75.1 mmol) in step 1 of Example 24, starting from intermediate S5 (4.2 g, 7.89 mmol) and stirred at 90 °C. MS: m / z = 518.2 (M+1, ESI+).

[0661] Step 2: N-(3-(cyclopropylethynyl)-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxy) Preparation of benzyl glycine

[0662] The product from step 1 (2.6 g, 5.02 mmol) was used to give N-(3-(cyclopropylethynyl)-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine (1.5 g, 64.71% yield) in the same manner as step 4 of Example 8. MS: m / z = 462.2 (M+1, ESI+).

[0663] Step 3: N-(6-amino-2,3-difluorophenyl)-2-((3-(cyclopropylethynyl)-6-morpholinylimidazo[1, Preparation of 2-b]pyridazine-8-yl)(4-methoxybenzyl)amino)acetamide

[0664] N-(6-amino-2,3-difluorophenyl)-2-((3-(cyclopropylethynyl)-6-morpholinylimidazo[1,2-b]pyridazin-8-yl)(4-methoxybenzyl)amino)acetamide (500 mg, yield 29.09%) was obtained by the same method as in step 5 of Example 8, except that the reaction time was changed to 16 hours. MS: m / z = 588.2 (M+1, ESI+).

[0665] Steps 4 and 5: 3-(cyclopropylethynyl)-N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-6-morpho Preparation of linylimidazo[1,2-b]pyridazine-8-amine

[0666] The compound of Example 25 (58 mg, yield 11.97%) was obtained as a white solid by the same method as steps 6 and 7 of Example 8, except that the reaction time was changed to 4 hours and 2 hours, respectively. 1 H NMR (400MHz, DMSO-d6) δ 7.62 (t, 1H), 7.53 (s, 1H), 7.28-7.16 (m, 2H), 6.05 (s, 1H), 4.81 (d, 2H), 3.68 (s, 4H), 3.34 (s, 4H), 1.66-1.62 (m, 1H), 0.95-0.92 (m,2H), 0.77-0.76 (m, 2H); MS: m / z = 450.4 (M+1, ESI+).

[0667] Example 26: N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-3-iodo-6-morpholinylimidazol[1] , Preparation of 2-b]pyridazine-8-amine

[0668]

[0669] Step 1: Preparation of 2-(chloromethyl)-6,7-difluoro-1H-benzo[d]imidazole

[0670] Intermediate B1 (10 g, 69.44 mmol) was dissolved in 6 M hydrochloric acid solution (100 mL), followed by the addition of 2-chloroacetic acid (9.84 g, 104.2 mmol) and stirring at 100 °C for 16 hours. Water (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give 2-(chloromethyl)-6,7-difluoro-1H-benzi[d]imidazole (10 g, 71.42% yield). MS: m / z = 203.2 (M+1, ESI+).

[0671] Step 2: 2-(chloromethyl)-6,7-difluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[] Preparation of [d]imidazolium

[0672] The product obtained in step 1 (10 g, 49.50 mmol) and DIEA (17.2 mL, 99 mmol) were dissolved in THF, and then TMSCl (11.85 g, 71.4 mmol) was added at 0 °C and the mixture was stirred at 25 °C for 16 hours. Water (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give 2-(chloromethyl)-6,7-difluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzi[d]imidazole (16 g, yield 97.56%). MS: m / z = 333.2 (M+1, ESI+).

[0673] Step 3: Preparation of N,N-bis(4-methoxybenzyl)-6-morpholinylimidazo[1,2-b]pyridazine-8-amine

[0674] Starting from intermediate S6 (4 g, 9.78 mmol), N,N-bis(4-methoxybenzyl)-6-morpholinylimidazo[1,2-b]pyridazine-8-amine (4 g, 88.98% yield) was obtained using the same method as in step 5 of Example 2. MS: m / z = 460.2 (M+1, ESI+).

[0675] Step 4: Preparation of N-(4-methoxybenzyl)-6-morpholinylimidazo[1,2-b]pyridazine-8-amine

[0676] The product obtained in step 3 (4 g, 9.78 mmol) was dissolved in DCM (20 mL), and then TFA (10 mL) was added. The mixture was stirred at 25 °C for 3 hours. 2 N sodium bicarbonate solution (100 mL) was added to the reaction mixture, and the product was extracted three times with DCM (40 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(4-methoxybenzyl)-6-morpholinylimidazo[1,2-b]pyridazine-8-amine (2.8 g, 84.33% yield). MS: m / z = 340.1 (M+1, ESI+).

[0677] Step 5: N-((6,7-difluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol- Preparation of 2-yl)methyl)-N-(4-methoxybenzyl)-6-morpholinylimidazo[1,2-b]pyridazine-8-amine

[0678] The product obtained in step 4 (2.8 g, 8.25 mmol) was dissolved in DMF (30 mL), and then NaH (990 mg, 24.75 mmol, 60% purity) was added in portions at 0 °C and stirred for 30 min. The product from step 2 (4.12 g, 12.38 mmol) was added to the reaction solution, and the mixture was stirred at 25 °C for 16 h. Water (300 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (300 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-((6,7-difluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-6-morpholinylimidazo[1,2-b]pyridazin-8-amine (2.6 g, yield 49.57%). MS: m / z = 636.5 (M+1, ESI+).

[0679] Step 6: N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-6-morpholinylimidazo[1,2-b]pyridyl Preparation of azinon-8-amine

[0680] The product obtained in step 5 (2.6 g, 4.09 mmol) was dissolved in DCM (20 mL), and then TFA (10 mL) was added. The mixture was stirred at 25 °C for 1 hour. 2 N sodium bicarbonate solution (200 mL) was added to the reaction mixture, and the product was extracted three times with DCM (60 mL). The organic layer was washed three times with brine (200 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-6-morpholinylimidazo[1,2-b]pyridazin-8-amine (1 g, yield 63.45%). MS: m / z = 386.2 (M+1, ESI+).

[0681] Step 7: N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-3-iodo-6-morpholinylimidazo[1,2-] b] Preparation of pyridazine-8-amine

[0682] The product obtained in step 6 (500 mg, 1.30 mmol) was dissolved in THF (10 mL), and then NIS (263 mg, 1.17 mmol) was added at 0 °C, and the mixture was stirred for 1 hour. Water (80 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (20 mL). The organic layer was washed three more times with sodium sulfite solution (80 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by preparative HPLC to give the compound of Example 26 (56 mg, yield 8.44%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.96 (d, 1H), 7.43 (d, 1H), 7.24-7.14 (m, 2H), 7.06 (t, 1H), 5.72 (d, 2H), 3.79 (t, 4H), 3.12 (t, 4H); MS: m / z = 512.0 (M+1, ESI+).

[0683] Example 27: Preparation of 3-chloro-N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-6-morpholinylimidazo[1,2-b]pyridazine-8-amine

[0684]

[0685] The product obtained in step 6 of Example 26 (300 mg, 778 μmol) was dissolved in THF (5 mL), and then NCS (135 mg, 1.01 mmol) was added at 0 °C and stirred for 1 hour. Water (50 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (15 mL). The organic layer was washed three more times with sodium sulfite solution (50 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give the compound of Example 27 (155 mg, yield 47.43%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.92 (d, 1H), 7.56 (t,1H), 7.42 (s, 1H), 7.25-7.13 (m, 2H), 5.69 (d, 2H), 3.78 (t, 4H), 3.17 (t,4H); MS: m / z = 421.7 (M+1, ESI+).

[0686] Example 28: Preparation of 6-(4-cyclopropylpiperazin-1-yl)-N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine

[0687]

[0688] Step 1: 6-(4-Cyclopropylpiperazin-1-yl)-N,N-bis(4-methoxybenzyl)-3-(trifluoromethyl)imidazo Preparation of [1,2-b]pyridazine-8-amine

[0689] Intermediate S7 (2.1 g, 4.52 mmol) and intermediate A15 (1.17 g, 6.77 mmol) were dissolved in DMSO (15 mL), and then TsOH (722 mg, 4.19 mmol) was added and the mixture was stirred at 140 °C for 16 hours. Water (150 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (50 mL). The organic layer was washed three times with brine (150 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give 6-(4-cyclopropylpiperazin-1-yl)-N,N-bis(4-methoxybenzyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (1.33 g, yield 55.97%). MS: m / z = 567.5 (M+1, ESI+).

[0690] Step 2: 6-(4-Cyclopropylpiperazin-1-yl)-N-(4-methoxybenzyl)-3-(trifluoromethyl)imidazo[1,2-] b] Preparation of pyridazine-8-amine

[0691] The product obtained in step 1 (800 mg, 1.41 mmol) was dissolved in DCM (6 mL) and TFA (2 mL), and then stirred at 25 °C for 3 hours. Sodium bicarbonate solution (100 mL) was added to the reaction mixture, and the product was extracted three times with DCM (40 mL). The organic layer was washed three times with brine (100 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give 6-(4-cyclopropylpiperazin-1-yl)-N-(4-methoxybenzyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (500 mg, yield 79.32%). MS: m / z = 447.2 (M+1, ESI+).

[0692] Step 3: 6-(4-Cyclopropylpiperazin-1-yl)-N-((6,7-Difluoro-1-((2-(trimethylsilyl)ethoxy) 2-(4-methoxybenzyl)-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-3-(trifluoromethyl)imidazol[1,2-b] Preparation of pyridazine-8-amine

[0693] 6-(4-cyclopropylpiperazin-1-yl)-N-((6,7-difluoro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (650 mg, yield 78.13%) was obtained by the same method as in step 5 of Example 26. MS: m / z = 743.5 (M+1, ESI+).

[0694] Step 4: 6-(4-cyclopropylpiperazin-1-yl)-N-((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)-3- Preparation of (trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0695] The product obtained in step 3 (650 mg, 875 μmol) was dissolved in TFA (10 mL) and stirred at 25 °C for 1 hour. The reaction mixture was concentrated and then purified by preparative HPLC to give the compound of Example 28 (140 mg, yield 32.49%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 12.70 (s, 1H), 7.87 (s,2H), 7.26-7.17 (m, 2H), 6.20 (s, 1H), 4.85 (d, 2H), 3.33 (s, 4H), 2.57 (s,4H), 1.60 (s, 1H), 0.42 (s, 2H), 0.33 (s, 2H); MS: m / z = 493.2 (M+1, ESI+).

[0696] In the following text, the compounds of Examples 29 to 67 were prepared according to Scheme II using appropriate intermediates from the intermediate series A in [Table 1] and the intermediate series B in [Table 3].

[0697] [Option II]

[0698]

[0699] In Scheme II, R 2 Having R with the compounds of each embodiment 2 The same structure is used, and X1, X2, and X3 also represent the substituent structures at corresponding positions in the compounds of each embodiment. The intermediates referred to as intermediates B1 to B30, described in the following [Table 3], have the structures shown in the following [Table 3], throughout the embodiments and Examples 29 to 67.

[0700] [Table 3]

[0701]

[0702] Example 29: Preparation of N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)-6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine

[0703]

[0704] Step 1: N-(4-methoxybenzyl)-N-(6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2- Preparation of [b]pyridazine-8-yl)glycine

[0705] Intermediate S8 (1 g, 2.26 mmol) and intermediate A2 (1.13 g, 11.29 mmol) were dissolved in 1,4-dioxane (20 mL), and then Ruphos-Pd-G3 (189 mg, 226 μmol), 2-dicyclohexylphosphino-2',6'-diisopropoxybiphenyl (RuPhos, 106 mg, 226 μmol), and cesium carbonate (3.68 g, 11.29 mmol) were added. The mixture was stirred at 110 °C for 16 hours. 1 N hydrochloric acid (150 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (50 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(4-methoxybenzyl)-N-(6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycine (400 mg, yield 37.02%). MS: m / z = 479.2 (M+1, ESI+).

[0706] Step 2: N-(2-amino-4,5-dichlorophenyl)-2-((4-methoxybenzyl)(6-(4-methylpiperazin-1-yl)- Preparation of 3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)amino)acetamide

[0707] The product obtained in step 1 (340 mg, 711 μmol) and intermediate B2 (629 mg, 3.55 mmol) were dissolved in DCM (10 mL), and then HOBT (480 mg, 3.55 mmol), EDCI (681 mg, 3.55 mmol), and DIEA (1.24 mL, 7.11 mmol) were added. The mixture was stirred at 25 °C for 16 hours. Water (80 mL) was added to the reaction mixture, and the product was extracted three times with DCM (30 mL). The organic layer was washed three times with brine (80 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(2-amino-4,5-dichlorophenyl)-2-((4-methoxybenzyl)(6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)amino)acetamide (200 mg, yield 44.15%). MS: m / z = 637.1 (M+1, ESI+).

[0708] Step 3: N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-6-(4-methyl) Preparation of pyridazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine

[0709] The product obtained in step 2 (150 mg, 235 μmol) was dissolved in TFA (8 mL), and the mixture was stirred at 70 °C for 16 hours. Sodium bicarbonate solution (100 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (40 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (130 mg, 89.19% yield). MS: m / z = 619.2 (M+1, ESI+).

[0710] Step 4: N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)-6-(4-methylpiperazin-1-yl)-3- Preparation of (trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0711] The product obtained in step 3 (139 mg, 204 μmol) was dissolved in DCM (4 mL), and then TFA (1 mL) was added. The mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated and then purified by preparative HPLC to give the compound of Example 29 (35 mg, yield 34.37%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.86 (s, 1H), 7.81 (t, 1H), 7.76 (s, 2H), 6.16 (s, 1H), 4.83 (d, 2H), 3.36 (t, 4H), 2.35 (t, 4H), 2.17 (s, 3H); MS: m / z = 499.1 (M+1, ESI+).

[0712] Example 30: Preparation of 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)azacyclobutane-3-carboxylonitrile

[0713]

[0714] Step 1: N-(6-(3-cyanozazine-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8- Preparation of ethyl glycine (N-(4-methoxybenzyl)glycine)

[0715] Intermediate S8 (2 g, 4.52 mmol) and intermediate A16 (742 mg, 9.03 mmol) were dissolved in 1,4-dioxane (20 mL), followed by the addition of Ruphos-Pd-G3 (189 mg, 226 μmol), 2-dicyclohexylphosphino-2',6'-diisopropoxybiphenyl (RuPhos, 106 mg, 226 μmol), and cesium carbonate (2.94 g, 9.03 mmol). The mixture was stirred at 110 °C for 48 hours. Water (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (60 mL). The organic layer was washed three times with brine (20 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(6-(3-cyanozycyclobutane-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine ethyl ester (1.5 g, yield 67.99%). MS: m / z = 489.1 (M+1, ESI+).

[0716] Step 2: N-(6-(3-cyanozazine-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8- Preparation of 4-(4-methoxybenzyl)glycine

[0717] The product obtained in step 1 (1.5 g, 3.07 mmol) was dissolved in THF (10 mL) and water (5 mL), and then lithium hydroxide (368 mg, 15.35 mmol) was added and the mixture was stirred at 25 °C for 2 hours. 1N hydrochloric acid (100 mL) was added to the reaction mixture, and the product was extracted three times with DCM (30 mL). The organic layer was washed three times with brine (100 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(6-(3-cyanozycyclobutane-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine (900 mg, yield 63.66%). MS: m / z = 461.1 (M+1, ESI+).

[0718] Step 3: N-(2-amino-4,5-dichlorophenyl)-2-((6-(3-cyanoazacyclobutane-1-yl)-3-(trifluoromethyl) Preparation of (-)imidazo[1,2-b]pyridazin-8-yl)(4-methoxybenzyl)amino)acetamide

[0719] The product obtained in step 2 (900 mg, 1.95 mmol) and intermediate B2 (1.38 g, 7.8 mmol) were dissolved in DCM (20 mL), and then HOBT (1.32 g, 9.77 mmol), EDCI (1.87 g, 9.77 mmol), and DIEA (3.41 mL, 19.55 mmol) were added. The mixture was stirred at 25 °C for 2 hours. Water (80 mL) was added to the reaction mixture, and the product was extracted three times with DCM (30 mL). The organic layer was washed three times with brine (80 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(2-amino-4,5-dichlorophenyl)-2-((6-(3-cyanoazacyclobutane-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)(4-methoxybenzyl)amino)acetamide (600 mg, yield 49.55%). MS: m / z = 619.2 (M+1, ESI+).

[0720] Step 4: 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)- Preparation of 3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)azacyclobutane-3-carboxylon

[0721] The product obtained in step 3 (600 mg, 969 μmol) was dissolved in AcOH (10 mL) and stirred at 100 °C for 1 hour. Sodium bicarbonate solution (100 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (40 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazol[1,2-b]pyridazin-6-yl)azacyclobutane-3-carboxylonitrile (400 mg, yield 68.66%). MS: m / z = 601.2 (M+1, ESI+).

[0722] Step 5: 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazolium Preparation of azido[1,2-b]pyridazine-6-yl)azacyclobutane-3-carboxylon

[0723] The product obtained in step 4 (400 mg, 665 μmol) was dissolved in DCM (6 mL), and then TFA (3 mL) was added. The mixture was stirred at 25 °C for 2 hours. The reaction mixture was concentrated and then purified by preparative HPLC to give the compound of Example 30 (55 mg, yield 17.18%) as a white solid. 1 H NMR (400 MHz, DMSO-d6)δ 12.61 (s, 1H), 7.98 (t, 1H), 7.89 (s, 1H), 7.77 (s, 2H), 5.71 (s, 1H), 4.81(d, 2H), 4.18 (t, 2H), 4.04 (dd, 2H), 3.85-3.79 (m, 1H); MS: m / z = 481.1 (M+1,ESI+).

[0724] Examples 31 to 56

[0725] The compounds of Examples 31 to 56 were prepared using intermediates A (corresponding to the structures of the target compounds in Table 1) and B (corresponding to the structures in Table 3), with appropriate changes to the amounts of reagents, catalysts and reaction conditions, in the same manner as in Example 30.

[0726] [Table 4]

[0727]

[0728]

[0729]

[0730]

[0731]

[0732]

[0733]

[0734] Example 57: Preparation of N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)-6-(piperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine

[0735]

[0736] The compound of Example 51 (650 mg, 1.23 mmol) was dissolved in 1,4-dioxane (10 mL), and then 6 M hydrochloric acid solution (2 mL) was added. The mixture was stirred at 100 °C for 48 hours. After concentrating the reaction mixture, the concentrate was purified by preparative HPLC to give the compound of Example 57 (120 mg, yield 20.06%) as a grayish-white solid. 1 H NMR(400 MHz, DMSO-d6) δ 7.86 (s, 1H), 7.82-7.77 (m, 3H), 6.13 (s, 1H), 4.83 (d,2H), 3.28 (s, 4H), 2.73 (s, 4H); MS: m / z = 485.0 (M+1, ESI+).

[0737] Example 58: Preparation of 6-(3-(cyclopropylamino)azacyclobutane-1-yl)-N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0738]

[0739] Steps 1 to 4: Cyclopropyl(1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl) Preparation of (-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-6-yl)azacyclobutane-3-yl)carbamate Preparation

[0740] Cyclopropyl (1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)azacyclobutane-3-yl)carbamate (450 mg, yield 32.90%) was obtained using the same method as steps 1 to 4 of Example 30. However, intermediate S8 (3 g, 6.77 mmol) and intermediate A36 (4.17 g, 16.94 mmol) were used in step 1 of Example 30 and the mixture was stirred for 16 hours, and the reaction time in step 3 of Example 30 was changed to 16 hours. MS: m / z = 765.3 (M+1, ESI+).

[0741] Step 5: 6-(3-(cyclopropylamino)azacyclobutane-1-yl)-N-((5,6-dichloro-1H-benzo[d]imidazole- Preparation of 2-yl)methyl)-N-(4-methoxybenzyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0742] The product obtained in step 4 (450 mg, 588 μmol) was dissolved in methanol (10 mL), and then Pd / C (45 mg) was added. The mixture was stirred at 25 °C for 3 hours under a H2 atmosphere. The reaction mixture was filtered and concentrated, and the concentrate was purified by silica gel chromatography to give 6-(3-(cyclopropylamino)azacyclobutane-1-yl)-N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (250 mg, yield 67.36%). MS: m / z = 631.2 (M+1, ESI+).

[0743] Step 6: 6-(3-(cyclopropylamino)azacyclobutane-1-yl)-N-((5,6-dichloro-1H-benzo[d]imidazole- Preparation of 2-yl)methyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0744] The compound of Example 58 (65 mg, yield 32.11%) was obtained as a white solid using the same method as step 5 of Example 30, except that the reaction time was changed to 1 hour.

[0745] 1H NMR (400 MHz, DMSO-d6) δ 12.63 (s, 1H), 7.86-7.83 (m, 2H), 7.77 (s,2H), 5.57 (s, 1H), 4.78 (d, 2H), 4.01 (t, 2H), 3.69-3.66 (m, 1H), 3.57 (t,2H), 2.91 (br s, 1H), 2.03-1.99 (m, 1H), 0.36-0.31 (m, 2H), 0.20-0.17 (m,2H); MS: m / z = 511.1 (M+1, ESI+).

[0746] Example 59: Preparation of 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)-3-hydroxyazacyclobutane-3-carboxylonitrile

[0747]

[0748] Steps 1 and 2: N-(6-(3-((tert-butyldimethylsilyl)oxy)azacyclobutane-1-yl)-3-(trifluoro) Preparation of (methyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine

[0749] N-(6-(3-((tert-butyldimethylsilyl)oxy)azacyclobutane-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine (2.5 g, crude) was obtained using the same method as steps 1 and 2 of Example 30. However, in step 1, intermediate S8 (5 g, 11.29 mmol) and intermediate A37 (5.29 g, 28.23 mmol) were used and the mixture was stirred for 16 hours, and in step 2, the reaction time was changed to 16 hours. MS: m / z = 566.2 (M+1, ESI+).

[0750] Step 3: N-(6-hydroxyazacyclobutane-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-yl)- Preparation of N-(4-methoxybenzyl)glycine

[0751] The product obtained in step 2 (1.5 g, 2.65 mmol) was dissolved in THF (20 mL), and then 1 M TBAF solution (10.6 mmol, 10.6 mL) was added. The mixture was stirred at 25 °C for 16 hours. Water (100 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (30 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give N-(6-hydroxyazacyclobutane-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine (700 mg, yield 58.48%). MS: m / z = 452.1 (M+1, ESI+).

[0752] Steps 4 and 5: 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino Preparation of (-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)azacyclobutane-3-ol

[0753] 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazol[1,2-b]pyridazin-6-yl)azacyclobutane-3-ol (780 mg, 90.31% yield) was obtained by the same method as steps 3 and 4 of Example 30. However, the reaction time in step 3 of Example 30 was changed to 16 hours, and the reaction temperature in step 4 of Example 30 was changed to 70°C and stirred for 3 hours. MS: m / z = 592.0 (M+1, ESI+).

[0754] Step 6: 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)- Preparation of 3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)azacyclobutane-3-one

[0755] The product obtained in step 5 (730 mg, 1.23 mmol) was dissolved in DCM (20 mL), and then Dess-Martin periodane (1.57 g, 3.70 mmol) was added. The mixture was stirred at 25 °C for 16 hours. Water (100 mL) was added to the reaction mixture, and the product was extracted three times with DCM (20 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)azacyclobutane-3-one (550 mg, yield 75.60%). MS: m / z = 590.0 (M+1, ESI+).

[0756] Step 7. 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)- Preparation of 3-(trifluoromethyl)imidazo[1,2-b]pyridazine-6-yl)-3-hydroxyazacyclobutane-3-carboxynitrile

[0757] The product obtained in step 6 (400 mg, 678 μmol) and sodium cyanide (166 mg, 3.39 mmol) were dissolved in THF / water (10 mL / 10 mL), and sodium bicarbonate (285 mg, 3.39 mmol) was added. The mixture was stirred at 25 °C for 16 hours. Water (100 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (30 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)-3-hydroxyazacyclobutane-3-carboxylonitrile (250 mg, yield 59.76%). MS:m / z = 617.1 (M+1, ESI+).

[0758] Step 8: 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazolium Preparation of azido[1,2-b]pyridazine-6-yl)-3-hydroxyazacyclobutane-3-carboxynitrile

[0759] The compound of Example 59 (30 mg, yield 14.90%) was obtained as a white solid by the same method as step 5 of Example 30, except that the reaction time was changed to 1 hour. 1 H NMR (400 MHz, DMSO-d6) δ 12.62(s, 1H), 8.00 (t, 1H), 7.90 (s, 1H), 7.77 (s, 2H), 7.53 (s, 1H), 5.78 (s,1H), 4.82 (d, 2H), 4.38 (d, 2H), 3.96 (d, 2H); MS: m / z = 497.1 (M+1, ESI+).

[0760] Example 60: Preparation of (1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)azacyclobutane-3-yl)methanol

[0761]

[0762] Steps 1 to 5: 6-(3-(((tert-butyldiphenylsilyl)oxy)methyl)azacyclobutane-1-yl)-N- Preparation of ((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0763] 6-(3-(((tert-butyldiphenylsilyl)oxy)methyl)azacyclobutane-1-yl)-N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (350 mg, yield 86.83%) was obtained by the same method as steps 1 to 5 of Example 30, except that intermediate S8 (2 g, 4.52 mmol) and intermediate A38 (7.35 g, 22.58 mmol) were used in step 1 and the mixture was stirred for 16 hours. MS: m / z = 724.2 (M+1, ESI+).

[0764] Step 6: (1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazolium Preparation of azido[1,2-b]pyridazine-6-yl)azacyclobutane-3-yl)methanol

[0765] The product obtained in step 5 (350 mg, 484 μmol) was dissolved in THF (10 mL), and then 1 M TBAF solution (2.42 mL, 2.42 mmol) was added. The mixture was stirred at 25 °C for 1 hour. Water (100 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (60 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by preparative HPLC to give the compound of Example 60 (100 mg, yield 42.61%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 12.60 (s, 1H), 7.84-7.77 (m, 4H), 5.56 (s, 1H), 4.79-4.73 (m, 3H), 3.87 (t, 2H), 3.62 (dd, 2H), 3.52 (t, 2H), 2.77-2.70 (m, 1H); MS: m / z = 486.0 (M+1, ESI+).

[0766] Example 61: Preparation of 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazol[1,2-b]pyridazin-6-yl)-3-hydroxypyrrolidine-3-carboxylonitrile

[0767]

[0768] Step 1: N-(6-(1,4-dioxa-7-azaspiro[4.4]non-7-yl)-3-(trifluoromethyl)imidazo[1,2- Preparation of ethyl glycine (b)pyridazine-8-yl)-N-(4-methoxybenzyl)glycine

[0769] N-(6-(1,4-dioxa-7-azaspiro[4.4]non-7-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine ethyl ester (780 mg, yield 17.92%) was obtained by the same method as in step 1 of Example 30, except that intermediate S8 (3.6 g, 8.13 mmol) and intermediate A39 (3.15 g, 24.29 mmol) were used and the mixture was stirred for 16 hours. MS: m / z = 535.2 (M+1, ESI+).

[0770] Step 2: N-(6-(1,4-dioxa-7-azaspiro[4.4]non-7-yl)-3-(trifluoromethyl)imidazo[1,2- Preparation of [b]pyridazine-8-yl)-N-(4-methoxybenzyl)glycine

[0771] The product obtained in step 1 (780 mg, 1.46 mmol) was dissolved in THF / water (10 mL / 5 mL), and lithium hydroxide (185 mg, 4.38 mmol) was added. The mixture was stirred at 50 °C for 2 hours. 0.5 N hydrochloric acid solution (80 mL) was added to the reaction mixture, and the product was extracted three times with DCM (20 mL). The organic layer was washed three times with brine (80 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(6-(1,4-dioxa-7-azaspiro[4.4]non-7-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine (410 mg, yield 55.25%). MS: m / z = 507.2 (M+1, ESI+).

[0772] Steps 3 to 5: 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl) Preparation of imidazo[1,2-b]pyridazin-6-yl)pyrrolidone-3-one

[0773] 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazol[1,2-b]pyridazin-6-yl)pyrrolidone-3-one (200 mg, yield 62.30%) was obtained by the same method as steps 3 to 5 of Example 30, except that the reaction times in steps 3 and 5 of Example 30 were changed to 16 hours and 1 hour, respectively. MS: m / z = 484.0 (M+1, ESI+).

[0774] Step 6: 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazolium Preparation of azido[1,2-b]pyridazine-6-yl)-3-hydroxypyrrolidine-3-carboxynitrile

[0775] The product obtained in step 5 (200 mg, 413 μmol) was used to obtain the compound of Example 61 (95 mg, 42.03% yield) as a white solid in the same manner as in step 7 of Example 59.1 H NMR (400 MHz, DMSO-d6) δ7.89 (t, 1H), 7.85 (s, 1H), 7.79 (s, 2H), 6.96 (s, 1H), 5.83 (s, 1H), 4.87(d, 2H), 3.74 (d, 1H), 3.64 (d, 1H), 3.54-3.49 (m, 1H), 3.44-3.38 (m, 1H), 2.46-2.39 (m, 1H), 2.33-2.28 (m, 1H); MS: m / z = 511.1 (M+1, ESI+).

[0776] Example 62: Preparation of (R)-1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)pyrrolidine-3-ol

[0777]

[0778] Step 1: (R)-N-(6-(3-((tert-butyldimethylsilyl)oxy)pyrrolidine-1-yl)-3-(trifluoromethyl) Preparation of ethyl glycine (4-(4-methoxybenzyl)glycine)-8-yl)-N-(4-methoxybenzyl)glycine

[0779] (R)-N-(6-(3-((tert-butyldimethylsilyl)oxy)pyrrolidine-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine ethyl ester (560 mg, yield 20.40%) was obtained by the same method as in step 1 of Example 30, except that intermediate S8 (2 g, 4.52 mmol) and intermediate A40 (2.73 g, 13.55 mmol) were used and the mixture was stirred for 16 hours. MS: m / z = 608.3 (M+1, ESI+).

[0780] Step 2: (R)-N-(6-(3-hydroxypyrrolidone-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8- Preparation of ethyl glycine (N-(4-methoxybenzyl)glycine)

[0781] The product obtained in step 1 (560 mg, 921 μmol) was dissolved in THF (5 mL), and then 1 M TBAF solution (1.84 mL, 1.84 mmol) was added. The mixture was stirred at 25 °C for 3 hours. A saturated ammonium chloride aqueous solution (50 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (10 mL). The organic layer was washed three times with brine (50 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give (R)-N-(6-(3-hydroxypyrrolidone-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine ethyl ester (400 mg, yield 87.97%). MS: m / z = 494.0 (M+1, ESI+).

[0782] Steps 3 to 6: (R)-1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoro) Preparation of (methyl)imidazo[1,2-b]pyridazin-6-yl)pyrrolidine-3-ol

[0783] The compound of Example 62 (56 mg, yield 25.93%) was obtained as a white solid by the same method as steps 2 to 5 of Example 30, except that the reaction times of steps 2, 3 and 5 of Example 30 were changed to 16 hours, 16 hours and 3 hours respectively, and the reaction temperature in step 4 was changed to 65°C and stirred for 5 hours. 1 H NMR (400MHz, DMSO-d6) δ 12.63 (s, 1H), 7.86 (s, 1H), 7.80-7.76 (m, 2H), 7.68 (s, 1H), 5.72 (s, 1H), 4.93 (d, 1H), 4.81 (d, 2H), 4.33 (s, 1H), 3.39-3.35 (m, 2H), 3.19 (d, 1H), 1.98-1.92 (m, 1H), 1.85-1.81 (m, 1H); MS: m / z = 486.2 (M+1, ESI+).

[0784] Example 63: Preparation of (S)-1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)pyrrolidine-3-ol

[0785]

[0786] Steps 1 to 4: (S)-6-(3-((tert-butyldimethylsilyl)oxy)pyrrolidine-1-yl)-N-((5,6-di) Chloro-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazine- Preparation of 8-amine

[0787] (S)-6-(3-((tert-butyldimethylsilyl)oxy)pyrrolidine-1-yl)-N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (850 mg, 87.12% yield) was obtained using the same method as steps 1 to 4 of Example 30. However, in step 1, intermediate S8 (3.1 g, 7.0 mmol) and intermediate A41 (2.82 g, 14.0 mmol) were used and stirred for 16 hours, and the reaction time in steps 2 and 3 was changed to 16 hours. MS: m / z = 720.2 (M+1, ESI+).

[0788] Step 5: (S)-1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino Preparation of 3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)pyrrolidine-3-ol

[0789] (S)-1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazol[1,2-b]pyridazin-6-yl)pyrrolidine-3-ol (600 mg, 83.89% yield) was obtained by the same method as step 2 of Example 62, except that the reaction time was changed to 16 hours. MS: m / z = 606.1 (M+1, ESI+).

[0790] Step 6: (S)-1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl) Preparation of (1,2-b)pyridine-6-yl)pyrrolidine-3-ol

[0791] The compound of Example 63 (290 mg, yield 59.86%) was obtained as a white solid using the same method as step 5 of Example 30, except that the reaction time was changed to 16 hours. 1 H NMR (400 MHz, DMSO-d6) δ 12.63(s, 1H), 7.80-7.76 (m, 4H), 5.72 (s, 1H), 4.93 (d, 1H), 4.81 (d, 2H), 4.33(s, 1H), 3.39-3.33 (m, 2H), 3.20 (d, 1H), 1.98-1.94 (m, 1H), 1.85-1.82 (m,1H); MS: m / z = 486.2 (M+1, ESI+).

[0792] Example 64: Preparation of 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)azacyclobutane-3-carboxylic acid

[0793]

[0794] Step 1: 1-(8-((2-ethoxy-2-oxoethyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazolium Preparation of [1,2-b]pyridazine-6-yl)azacyclobutane-3-carboxylic acid tert-butyl ester

[0795] 1-(8-((2-ethoxy-2-oxoethyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)azacyclobutane-3-carboxylic acid tert-butyl ester (1 g, yield 39.29%) was obtained by the same method as in step 1 of Example 30, except that intermediate S8 (2 g, 4.52 mmol) and intermediate A42 (3.55 g, 22.58 mmol) were stirred for 16 hours. MS: m / z = 564.2 (M+1, ESI+).

[0796] Step 2: N-(6-(3-(tert-butoxycarbonyl)azacyclobutane-1-yl)-3-(trifluoromethyl)imidazo[1,2-b] Preparation of pyridazine-8-yl)-N-(4-methoxybenzyl)glycine

[0797] The product obtained in step 1 (1 g, 1.77 mmol) was dissolved in 1,2-dichloroethane (20 mL), and then trimethyltin hydroxide (1.60 g, 8.87 mmol) was added. The mixture was stirred at 70 °C for 16 hours. 0.5 N hydrochloric acid solution (100 mL) was added to the reaction mixture, and the mixture was extracted three times with DCM (30 mL). The organic layer was washed three times with brine (100 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give N-(6-(3-(tert-butoxycarbonyl)azacyclobutane-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine (750 mg, 78.93% yield). MS: m / z = 536.1 (M+1, ESI+).

[0798] Steps 3 to 5: 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl) Preparation of imidazo[1,2-b]pyridazin-6-yl)azacyclobutane-3-carboxylic acid

[0799] The compound of Example 64 (70 mg, yield 31.56%) was obtained as a white solid by the same method as steps 3 to 5 of Example 30, except that the reaction time in step 5 of Example 30 was changed to 1 hour. 1 H NMR (400MHz, DMSO-d6) δ 7.90 (t, 1H), 7.85 (s, 1H), 7.77 (s, 2H), 5.65 (s, 1H), 4.80(d, 2H), 4.05 (t, 2H), 3.93 (t, 2H), 3.48-3.41 (m, 1H); MS: m / z = 500.0 (M+1,ESI+).

[0800] Example 65: Preparation of 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazol[1,2-b]pyridazin-6-yl)azacyclobutane-3-carboxamide

[0801]

[0802] Step 1: 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)- Preparation of 3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)azacyclobutane-3-carboxylic acid

[0803] The product obtained in step 4 of Example 64 (310 mg, 458 μmol) was dissolved in 1,4-dioxane (8 mL) and water (4 mL), and then sodium hydroxide (92 mg, 2.29 mmol) was added. The mixture was then stirred at 100 °C for 16 hours. 1 N hydrochloric acid solution (80 mL) was added to the reaction mixture, and the product was extracted three times with DCM (30 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)azacyclobutane-3-carboxylic acid (250 mg, 87.94% yield). MS: m / z = 620.1 (M+1,ESI+).

[0804] Step 2: 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)- Preparation of 3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)azacyclobutane-3-carboxamide

[0805] 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)azacyclobutane-3-carboxamide (150 mg, 60.10% yield) was obtained by the same method as in step 3 of Example 30, except that ammonium chloride (108 mg, 2.01 mmol) was used instead of intermediate B2 (1.38 g, 7.8 mmol). MS: m / z = 619.2 (M+1, ESI+).

[0806] Step 3: 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazolium Preparation of azido[1,2-b]pyridazine-6-yl)azacyclobutane-3-carboxamide

[0807] The compound of Example 65 (50 mg, yield 41.35%) was obtained as a white solid by the same method as step 5 of Example 30, except that the reaction time was changed to 1 hour. 1H NMR (400 MHz, DMSO-d6) δ 12.61(s, 1H), 7.90-7.76 (m, 4H), 7.45 (s, 1H), 7.01 (s, 1H), 5.62 (s, 1H), 4.80(d, 2H), 3.98 (t, 2H), 3.89 (t, 2H), 3.42-3.36 (m, 1H); MS: m / z = 499.0 (M+1,ESI+).

[0808] Example 66: Preparation of 4-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)piperazine-1-carboxamide

[0809]

[0810] Step 1: 4-(8-((2-ethoxy-2-oxoethyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazolium Preparation of [1,2-b]pyridazine-6-yl)piperazine-1-carboxylic acid benzyl ester

[0811] 4-(8-((2-ethoxy-2-oxoethyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)piperazine-1-carboxylic acid benzyl ester (700 mg, yield 24.73%) was obtained by the same method as in step 1 of Example 30, except that intermediate S8 (2 g, 4.52 mmol) and intermediate A11 (4.97 g, 22.58 mmol) were stirred for 16 hours. MS: m / z = 627.3 (M+1, ESI+).

[0812] Step 2: N-(4-methoxybenzyl)-N-(6-(piperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridyl Preparation of azinon-8-yl)glycine ethyl ester

[0813] The product obtained in step 1 (700 mg, 1.12 mmol) was dissolved in methanol (15 mL), and then Pd / C (100 mg) was added. The mixture was then stirred at 25 °C for 48 hours under hydrogen ignition. The reaction mixture was filtered and concentrated, and the concentrate was purified by silica gel chromatography to give N-(4-methoxybenzyl)-N-(6-(piperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycine ethyl ester (500 mg, yield 90.88%). MS: m / z = 493.2 (M+1, ESI+).

[0814] Step 3: N-(6-(4-carbamoylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8- Preparation of ethyl glycine (N-(4-methoxybenzyl)glycine)

[0815] The product obtained in step 2 (500 mg, 1.02 mmol) and trimethoxysilyl isocyanate (351 mg, 3.05 mmol) were dissolved in THF (10 mL), and then DIEA (0.53 mL, 3.05 mmol) was added. The mixture was stirred at 25 °C for 4 hours. Water (80 mL) was added to the reaction mixture, and the product was extracted three times with DCM (30 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give N-(6-(4-carbamoylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine ethyl ester (450 mg, yield 82.77%). MS: m / z = 536.3 (M+1, ESI+).

[0816] Step 4: N-(6-(4-carbamoylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8- Preparation of 4-(4-methoxybenzyl)glycine

[0817] N-(6-(4-carbamoylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine (280 mg, yield 65.66%) was obtained by the same method as in step 2 of Example 64. MS: m / z = 508.2 (M+1, ESI+).

[0818] Steps 5 to 7: 4-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl) Preparation of imidazo[1,2-b]pyridazine-6-yl)piperazine-1-carboxamide

[0819] The compound of Example 66 (39 mg, yield 28.16%) was obtained as a white solid by the same method as steps 3 to 5 of Example 30, except that the reaction temperature in step 4 of Example 30 was changed to 70°C and stirred for 3 hours, and the reaction time in step 5 was changed to 1 hour. 1 H NMR (400 MHz, DMSO-d6) δ 12.58 (s, 1H), 7.88 (s, 1H), 7.84 (t, 1H), 7.77 (s, 2H), 6.22 (s, 1H), 6.05 (s, 2H), 4.84(d, 2H), 3.35 (s, 8H); MS: m / z = 528.1 (M+1, ESI+).

[0820] Example 67: Preparation of 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)piperidine-4-carboxamide

[0821]

[0822] Step 1: 1-(8-((2-ethoxy-2-oxoethyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazolium Preparation of [1,2-b]pyridazine-6-yl)piperidine-4-carboxylic acid tert-butyl ester

[0823] Intermediate S8 (1.1 g, 2.48 mmol) was dissolved in intermediate A43 (15 mL) and stirred at 120 °C for 16 hours. Water (100 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (30 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give tert-butyl 1-(8-((2-ethoxy-2-oxoethyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)piperidine-4-carboxylate (1.05 g, 71.42%). MS: m / z = 592.1 (M+1, ESI+).

[0824] Steps 2 to 5: 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl) Preparation of imidazo[1,2-b]pyridazin-6-yl)piperidine-4-carboxylic acid

[0825] 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazol[1,2-b]pyridazin-6-yl)piperidine-4-carboxylic acid (200 mg, yield 76.05%) was obtained using the same method as steps 2 to 5 of Example 30, except that the reaction times in steps 2, 3, and 5 of Example 30 were changed to 1 hour, 16 hours, and 1 hour, respectively, and the reaction temperature in step 4 was changed to 70°C with stirring for 4 hours. MS: m / z = 528.2 (M+1, ESI+).

[0826] Step 6: 1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazolium Preparation of zo[1,2-b]pyridazine-6-yl)piperidine-4-carboxamide

[0827] The compound of Example 67 (10 mg, yield 5.00%) was obtained as a white solid by the same method as step 3 of Example 30, except that ammonium chloride (102 mg, 1.90 mmol) was used instead of intermediate B2 (1.38 g, 7.8 mmol) and the mixture was stirred for 16 hours. 1H NMR (400 MHz, DMSO-d6) δ 12.56 (s, 1H), 7.85 (s,1H), 7.79-7.72 (m, 3H), 7.24 (d, 1H), 6.76 (s, 1H), 6.16 (s, 1H), 4.83 (d,2H), 4.06-4.01 (m, 2H), 2.82-2.67 (m, 2H), 2.44-2.28 (m, 1H), 1.71-1.66 (m,2H), 1.53-1.44 (m, 2H);MS: m / z = 527.1 (M+1, ESI+) 。

[0828] In the following text, the compounds of Examples 68 to 81 were prepared according to Scheme III using appropriate intermediates from the intermediate series B in [Table 3].

[0829] [Option III]

[0830]

[0831] (In Scheme III, X) 3a X 4a X 5a and X 6a No more than one of them is N, and the rest are CR. 3d , where each R 3d Independently H, halogen, cyano, hydroxyl, C1-C6 alkoxy, or C1-C6 alkyl, provided that R 3d Any C1-C6 alkoxy and C1-C6 alkyl groups may optionally be substituted with a halogen, cyano, or hydroxyl group.

[0832] Example 68: Preparation of N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0833]

[0834] Step 1: N-(2-amino-4,5-dichlorophenyl)-2-((4-methoxybenzyl)(6-morpholino-3-(trifluoromethyl)) Preparation of imidazo[1,2-b]pyridazine-8-yl)amino)acetamide

[0835] N-(2-amino-4,5-dichlorophenyl)-2-((4-methoxybenzyl)(6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)amino)acetamide (1.7 g, yield 66.69%) was obtained using the same method as step 3 of Example 30, except that intermediate S9 (1.9 g, 4.08 mmol) was used as the starting material and the reaction time was changed to 16 hours. MS: m / z = 624.2 (M +1, ESI+).

[0836] Step 2: N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-6-morpholine Preparation of 3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0837] N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (1.6 g, 96.91% yield) was obtained by the same method as in step 4 of Example 30, except that the reaction temperature was changed to 70 °C and the mixture was stirred for 16 hours. MS: m / z = 606.2 (M +1, ESI+).

[0838] Step 3: N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)-6-morpholino-3-(trifluoromethyl)imidazolium Preparation of zo[1,2-b]pyridazine-8-amine

[0839] The compound of Example 68 (720 mg, yield 56.12%) was obtained as a white solid by the same method as step 5 of Example 30. 1 H NMR (400 MHz, DMSO-d6) δ 7.88-7.77 (m, 4H), 6.18 (s, 1H), 4.84(d, 2H), 3.67 (t, 4H), 3.34 (t, 4H); MS: m / z = 486.1 (M+1, ESI+).

[0840] Examples 69 to 81

[0841] The compounds of Examples 69 to 81 were prepared by using intermediates from the intermediate B series corresponding to the structure of the target compound, and by appropriately changing the amounts of reagents, catalysts and reaction conditions, in a manner similar to that of Example 68.

[0842] [Table 5]

[0843]

[0844]

[0845]

[0846]

[0847] Example 82: Preparation of N-(imidazo[1,2-a]pyridin-2-ylmethyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0848]

[0849] Step 1: Preparation of 2-(chloromethyl)imidazo[1,2-a]pyridine

[0850] Pyridine-2-amine (2.2 g, 23.38 mmol) and 1,3-dichloroprop-2-one (2.97 g, 23.38 mmol) were dissolved in ethanol (30 mL), and the mixture was stirred at 90 °C for 16 hours. The reaction mixture was concentrated, and the concentrate was purified by silica gel chromatography to give 2-(chloromethyl)imidazo[1,2-a]pyridine (400 mg, yield 10.27%). MS: m / z = 167.0 (M+1, ESI+).

[0851] Step 2: N-(imidazo[1,2-a]pyridin-2-ylmethyl)-N-(4-methoxybenzyl)-6-morpholino-3-(tri-) Preparation of fluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0852] The product obtained in step 1 (245 mg, 1.47 mmol) and the product obtained in step 2 (400 mg, 982 μmol) used to prepare intermediate S9 were dissolved in DMF (10 mL), and cesium carbonate (960 mg, 2.95 mmol) was added. The mixture was then stirred at 90 °C for 3 hours. A saturated aqueous solution of ammonium chloride (100 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (30 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give N-(imidazo[1,2-a]pyridin-2-ylmethyl)-N-(4-methoxybenzyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (270 mg, yield 51.16%). MS: m / z = 538.3 (M+1, ESI+).

[0853] Step 3: N-(imidazo[1,2-a]pyridin-2-ylmethyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2- b] Preparation of pyridazine-8-amine

[0854] The product obtained in step 2 (270 mg, 502.3 μmol) was dissolved in DCM (6 mL), and then TFA (3 mL) was added. The mixture was then stirred at 25 °C for 3 hours. The reaction mixture was concentrated, and the concentrate was purified by silica gel chromatography to give the compound of Example 82 (90 mg, yield 42.93%) as a white solid.1 H NMR (400 MHz, DMSO-d6) δ 8.48 (d, 1H), 7.84 (d, 2H), 7.74 (t, 1H), 7.50 (d, 1H), 7.23-7.19 (m,1H), 6.87-6.83 (m, 1H), 6.20 (s, 1H), 4.68 (d, 2H), 3.68 (t, 4H), 3.36 (t,4H); MS: m / z = 418.2 (M+1, ESI+).

[0855] Example 83: Preparation of N-(imidazo[1,2-a]pyrimidin-2-ylmethyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0856]

[0857] Step 1: Preparation of 2-(chloromethyl)imidazo[1,2-a]pyrimidine

[0858] Pyrimidine-2-amine (5 g, 52.57 mmol) was dissolved in diethylene glycol dimethyl ether (20 mL), and then 1,3-dichloroprop-2-one (20.03 g, 157.72 mmol) was added. The mixture was stirred at 70 °C for 16 hours. Water (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (50 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give 2-(chloromethyl)imidazo[1,2-a]pyrimidine (2.5 g, yield 28.37%). MS: m / z = 167.8 (M+1, ESI+).

[0859] Step 2: N-(imidazo[1,2-a]pyrimidin-2-ylmethyl)-N-(4-methoxybenzyl)-6-morpholino-3-(tri-) Preparation of fluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0860] The product obtained in step 2 (1 g, 2.45 mmol) used to prepare intermediate S9 was dissolved in DMF (5 mL), and then NaH (295 mg, 7.36 mmol, 60% purity) was added to it at 0 °C, followed by stirring for 30 min. The product from step 1 (411 mg, 2.45 mmol) was added to the reaction solution, and the mixture was stirred at 25 °C for 3 h. Ice water (50 mL) was added to the reaction mixture, and the product was washed three times with EtOAc (10 mL). The organic layer was washed three times with brine (50 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give N-(imidazo[1,2-a]pyrimidin-2-ylmethyl)-N-(4-methoxybenzyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (680 mg, yield 51.44%). MS: m / z = 539.3 (M+1, ESI+).

[0861] Step 3: N-(imidazo[1,2-a]pyrimidin-2-ylmethyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-] b] Preparation of pyridazine-8-amine

[0862] The product obtained in step 2 (680 mg, 1.26 mmol) was dissolved in DCM (10 mL), and then TFA (5 mL) was added. The mixture was then stirred at 25 °C for 16 hours. The reaction mixture was concentrated, and the concentrate was purified by preparative HPLC to give the compound of Example 83 (70 mg, yield 13.25%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ8.89 (dd, 1H), 8.49 (dd, 1H), 7.86 (s, 1H), 7.81 (t, 1H), 7.78 (s, 1H), 7.02(dd, 1H), 6.18 (s, 1H), 4.72 (d, 2H), 3.68 (t, 4H), 3.35 (t, 4H); MS: m / z =419.1 (M+1, ESI+).

[0863] Example 84: Preparation of N-((1H-pyrrolo[2,3-b]pyridin-2-yl)methyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine

[0864]

[0865] Step 1: 1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-2-carboxylic acid Preparation of methyl ester

[0866] 1H-pyrrolo[2,3-b]pyridine-2-carboxylate methyl ester (3.2 g, 18.16 mmol) was dissolved in DMF (50 mL), and then NaH (1.09 g, 27.25 mmol, 60% purity) was added in portions at 0 °C, followed by stirring at 25 °C for 1 hour. 2-(trimethylsilyl)ethoxymethyl chloride (3.86 mL, 21.80 mmol) was added to the reaction solution, and the mixture was stirred at 25 °C for 16 hours. A saturated aqueous solution of ammonium chloride (500 mL) was added to the reaction mixture, and the product was washed three times with EtOAc (100 mL). The organic layer was washed with brine (100 mL), dried over Na₂SO₄, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give methyl 1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine-2-carboxylate (2.8 g, yield 50.31%). MS: m / z = 307.1 (M+1, ESI+).

[0867] Step 2: (1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-2-yl) Preparation of methanol

[0868] The product obtained in step 1 (2.8 g, 9.14 mmol) was dissolved in THF (50 mL) and methanol (10 mL), and sodium borohydride (491 mg, 13.71 mmol) was added. The mixture was then stirred at 50 °C for 5 hours. A saturated aqueous solution of ammonium chloride (200 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (60 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give (1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)methanol (2.1 g, 82.55% yield). MS: m / z = 279.1 (M+1, ESI+).

[0869] Step 3: 2-(chloromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b] Preparation of pyridine

[0870] The product obtained in step 2 (2.1 g, 7.54 mmol) was dissolved in DCM (15 mL), and thionyl chloride (1.64 mL, 22.63 mmol) was slowly added to it at 0 °C, followed by stirring at 25 °C for 3 hours. The reaction mixture was concentrated, and the concentrate was then purified by silica gel chromatography to give 2-(chloromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-b]pyridine (1.5 g, yield 66.99%). MS: m / z = 297.1 (M+1, ESI+).

[0871] Steps 4 and 5: N-((1H-pyrrolo[2,3-b]pyridin-2-yl)methyl)-6-morpholino-3-(trifluoromethyl)imidazolium Preparation of zo[1,2-b]pyridazine-8-amine

[0872] The compound of Example 84 (60 mg, yield 34.28%) was obtained as a white solid by the same method as steps 2 and 3 of Example 82, except that the reaction time in step 2 of Example 82 was changed to 5 hours. 1 H NMR (400MHz, DMSO-d6) δ 11.55 (s, 1H), 8.13 (dd, 1H), 7.86-7.83 (m, 2H), 7.70 (t,1H), 7.00 (dd, 1H), 6.36 (s, 1H), 6.18 (s, 1H), 4.74 (d, 2H), 3.68 (t, 4H), 3.36 (t, 4H); MS: m / z = 418.3 (M+1, ESI+).

[0873] Example 85: Preparation of N-(imidazo[1,2-b]pyridazin-2-ylmethyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine

[0874]

[0875] The compound of Example 85 (70 mg, yield 20.02%) was obtained as a white solid by the same method as steps 1 to 3 of Example 82. However, in step 1 of Example 82, pyridazine-3-amine (5.2 g, 54.68 mmol) was used as the starting material, the mixture was stirred at 85°C for 3 hours, and the reaction time in step 2 was changed to 5 hours. 1H NMR (400 MHz, DMSO-d6) δ 8.48 (dd, 1H), 8.21 (s, 1H), 8.07 (dd, 1H), 7.85 (s, 1H), 7.76 (t,1H), 7.21 (dd, 1H), 6.21 (s, 1H), 4.72 (d, 2H), 3.69 (t, 4H), 3.36 (t, 4H); MS: m / z = 419.0 (M+1, ESI+).

[0876] Example 86: Preparation of 6-morpholino-N-(pyrazolo[1,5-a]pyridin-2-ylmethyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0877]

[0878] Step 1: Preparation of methyl pyrazolo[1,5-a]pyridine-2-carboxylate

[0879] Pyrazolo[1,5-a]pyridine-2-carboxylic acid (1.1 g, 6.78 mmol) was dissolved in methanol (30 mL), and then concentrated sulfuric acid (0.11 mL, 2.04 mmol) was added. The mixture was then stirred at 75 °C for 5 hours. After concentrating the reaction mixture, sodium bicarbonate solution (100 mL) was added, and the product was washed three times with EtOAc (30 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give methyl pyrazolo[1,5-a]pyridine-2-carboxylate (950 mg, yield 79.49%). MS: m / z = 177.1 (M+1, ESI+).

[0880] Steps 2 to 5: 6-morpholino-N-(pyrazolo[1,5-a]pyridin-2-ylmethyl)-3-(trifluoromethyl)imidazo Preparation of [1,2-b]pyridazine-8-amine

[0881] The compound of Example 86 (100 mg, yield 41.54%) was obtained as a white solid by the same method as steps 2 to 5 of Example 84, except that the reaction times in steps 3 and 4 of Example 84 were changed to 2 hours and 3 hours, respectively. 1H NMR (400 MHz, DMSO-d6) δ 8.62 (dd, 1H), 7.85-7.82 (m, 2H), 7.60 (d,1H), 7.20-7.15 (m, 1H), 6.86-6.82 (m, 1H), 6.51 (s, 1H), 6.18 (s, 1H), 4.73(d, 2H), 3.68 (t, 4H), 3.34 (t, 4H); MS: m / z = 418.1 (M+1, ESI+).

[0882] Example 87: Preparation of N-((1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine

[0883]

[0884] The compound of Example 87 (74 mg, yield 34.26%) was obtained as a white solid by the same method as steps 1 to 5 of Example 84. 1 H NMR (400 MHz, DMSO-d6) δ 11.19 (s, 1H), 8.24 (d, 1H), 7.86 (s, 1H), 7.79 (s, 1H), 7.69 (d, 1H), 7.05-7.00 (m, 1H), 6.51 (s, 1H), 6.16 (d, 1H), 4.78 (s, 2H), 3.77-3.62 (m, 4H), 3.40-3.24 (m, 4H); MS: m / z =418.1 (M+1, ESI+).

[0885] Example 88: Preparation of N-((5,7-dimethylimidazo[1,2-a]pyrimidin-2-yl)methyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0886]

[0887] The compound of Example 88 (28 mg, yield 10.14%) was obtained as a white solid using the same method as steps 1 to 3 of Example 82. However, in step 1 of Example 82, 4,6-dimethyl-4,5-dihydropyrimidine-2-amine (6 g, 50 mmol) was dissolved in DME (80 mL) and stirred at 50 °C; in step 2, the mixture was stirred at 25 °C for 2 hours using NaH (145 mg, 6.03 mmol, 60% purity); and in step 3, the reaction time was changed to 1 hour. 1 H NMR (400MHz, DMSO-d6) δ 7.85 (s, 1H), 7.73-7.67 (m, 1H), 7.66 (s, 1H), 6.84 (s, 1H), 6.19 (s, 1H), 4.68 (d, 2H), 3.72-3.64 (m, 4H), 3.38-3.33 (m, 4H), 2.56 (s,3H), 2.48 (s, 3H); MS: m / z = 447.2 (M+1, ESI+).

[0888] Example 89: Preparation of 6-morpholino-N-(pyrazolo[1,5-a]pyrimidin-2-ylmethyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0889]

[0890] Step 1: Preparation of (5-nitro-1H-pyrazole-3-yl)methanol

[0891] Methyl 5-nitro-1H-pyrazole-3-carboxylate (9 g, 52.60 mmol) was dissolved in THF (200 mL), and then lithium borohydride (LiB2) was added in portions over 1 hour at 0 °C. H 4, 5.73 g (263 mmol) were added, and the mixture was stirred at 40 °C for 48 hours. The reaction mixture was slowly cooled to room temperature by pouring in ice water (300 mL), and the product was extracted three times with EtOAc (60 mL). The organic layer was washed three times with brine (300 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give (5-nitro-1H-pyrazole-3-yl)methanol (4.5 g, yield 59.84%). MS: m / z = 144.1 (M+1, ESI+).

[0892] Step 2: Preparation of (5-amino-1H-pyrazole-3-yl)methanol

[0893] The product obtained in step 1 (4.50 g, 31.45 mmol) was dissolved in methanol (50 mL), and then Pd / C (1 g) was added. The mixture was then stirred at 25 °C for 16 hours under a H2 atmosphere. The reaction mixture was filtered and concentrated to give (5-amino-1H-pyrazole-3-yl)methanol (3.5 g, yield 98.39%). MS: m / z = 113.9 (M+1, ESI+).

[0894] Step 3: Preparation of pyrazolo[1,5-a]pyrimidin-2-ylmethanol

[0895] The product obtained in step 2 (3.5 g, 30.94 mmol) and 1,1,3,3-tetramethoxypropane (6.10 g, 37.13 mmol) were dissolved in ethanol (50 mL), and then TsOH (2.66 g, 15.47 mmol) was added. The mixture was then stirred at 80 °C for 1 hour. The reaction mixture was concentrated, and the concentrate was purified by silica gel chromatography to give pyrazolo[1,5-a]pyrimidin-2-ylmethanol (360 mg, yield 7.80%). MS: m / z = 150.2 (M+1, ESI+).

[0896] Step 4: Preparation of 2-(chloromethyl)pyrazolo[1,5-a]pyrimidine

[0897] The product obtained in step 3 (360 mg, 2.41 mmol) was dissolved in toluene (5 mL), and then thionyl chloride (861 mg, 7.24 mmol, 0.53 mL) was added. The mixture was then stirred at 50 °C for 1 hour. The reaction mixture was concentrated to give 2-(chloromethyl)pyrazolo[1,5-a]pyrimidine (360 mg, 2.15 mmol, 88.99% yield). MS: m / z = 168.0 (M+1, ESI+).

[0898] Steps 5 and 6: 6-morpholino-N-(pyrazolo[1,5-a]pyrimidin-2-ylmethyl)-3-(trifluoromethyl)imidazo Preparation of [1,2-b]pyridazine-8-amine

[0899] The compound of Example 89 (290 mg, yield 31.12%) was obtained as a white solid by the same method as steps 4 and 5 of Example 84. 1H NMR (400 MHz, DMSO-d6) δ 9.09-9.06 (m, 1H), 8.51 (dd, 1H), 7.89-7.85 (m, 2H), 7.00 (dd, 1H), 6.64 (s, 1H), 6.17 (s, 1H), 4.77 (d, 2H), 3.68 (t, 4H), 3.35 (t, 4H); MS: m / z = 419.2 (M+1, ESI+).

[0900] Example 90: Preparation of N-((5,6-dichloro-1H-indol-2-yl)methyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0901]

[0902] Step 1: Preparation of 6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0903] The product from step 1 (36 g, 68.31 mmol) used to prepare intermediate S9 was dissolved in DCM (300 mL), and then TFA (300 mL) was added. The mixture was then stirred at 25 °C for 16 hours. Sodium bicarbonate solution (2000 mL) was added to the reaction mixture, and the product was extracted three times with DCM (300 mL). The organic layer was washed three times with brine (2000 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give 18 g (91.83%) of 6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine as a pale yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ7.85 (s, 1H), 6.90 (br s, 2H), 6.09 (s, 1H), 3.71 (t, 4H), 3.33 (t, 4H). MS: m / z = 288.1 (M+1, ESI+).

[0904] Step 2: N-((5,6-dichloro-1-(benzenesulfonyl)-1H-indol-2-yl)methyl)-6-morpholino-3-(trifluoromethyl) Preparation of (1,2-b)imidazo[1,2-b]pyridazine-8-amine

[0905] The product obtained in step 1 (1.22 g, 4.23 mmol) and intermediate B20 (3 g, 8.47 mmol) were dissolved in THF (50 mL), and then Ti(OEt)4 (2.90 g, 12.70 mmol) was added. The mixture was then stirred at 90 °C for 16 hours. Afterward, the reaction mixture was cooled to 0 °C, and NaBH4 (641 mg, 16.94 mmol) was added, followed by stirring at 0 °C for 1 hour. Water (400 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (80 mL). The organic layer was washed three times with brine (400 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give N-((5,6-dichloro-1-(benzenesulfonyl)-1H-indol-2-yl)methyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine (250 mg, crude) as a yellow solid. MS: m / z = 625.2 (M+1, ESI+).

[0906] Step 3: N-((5,6-dichloro-1H-indol-2-yl)methyl)-6-morpholino-3-(trifluoromethyl)imidazo[1, Preparation of 2-b]pyridazine-8-amine

[0907] The product obtained in step 2 (250 mg, crude) was dissolved in THF (10 mL), and then 4 N NaOH aqueous solution (10 mL) was added. The mixture was then stirred at 90 °C for 16 hours. After cooling the reaction mixture to room temperature, water (20 mL) was added, and the mixture was extracted three times with DCM (5 mL). The organic layer was washed three times with brine (20 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give N-((5,6-dichloro-1H-indol-2-yl)methyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine (30 mg, yield 15.47%) as a pale yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 11.27 (s, 1H), 7.86 (s, 1H), 7.78 (t, 1H), 7.71(s, 1H), 7.59 (s, 1H), 6.40 (s, 1H), 6.15 (s, 1H), 4.73 (d, 2H), 3.68 (t,4H), 3.60-3.34 (m, 4H); MS: m / z = 485.1 (M+1, ESI+).

[0908] Example 91: Preparation of N-((5,6-dichlorobenzo[d]oxazol-2-yl)methyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0909]

[0910] Step 1: N-(4,5-dichloro-2-hydroxyphenyl)-2-((4-methoxybenzyl)(6-morpholino-3-(trifluoromethyl)) Preparation of imidazo[1,2-b]pyridazine-8-yl)amino)acetamide

[0911] N-(4,5-dichloro-2-hydroxyphenyl)-2-((4-methoxybenzyl)(6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)amino)acetamide (1.3 g, 48.37% yield) was obtained as a yellow solid using the same method as in step 3 of Example 30, except that intermediate S9 (2 g, 4.34 mmol) and 2-amino-4,5-dichlorophenol (1.91 g, 10.74 mmol) were used and the mixture was stirred for 16 hours. MS: m / z = 625.1 (M+1, ESI+).

[0912] Step 2: N-((5,6-dichlorobenzo[d]oxazol-2-yl)methyl)-6-morpholino-3-(trifluoromethyl)imidazo Preparation of [1,2-b]pyridazine-8-amine

[0913] The product obtained in step 1 (1.3 g, 2.08 mmol) was dissolved in toluene (15 mL), and then TsOH (1.79 g, 10.39 mmol) was added. The mixture was then stirred at 120 °C for 2 hours. The reaction mixture was cooled to room temperature, and ice water (200 mL) was added. The product was extracted three times with EtOAc (50 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the compound of Example 91 (30 mg, yield 2.68%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.22 (s, 1H),8.09 (s, 1H), 7.94 (t, 1H), 7.88 (s, 1H), 6.31 (s, 1H), 4.97 (d, 2H), 3.68(t, 4H), 3.37 (t, 4H); MS: m / z = 487.1 (M+1, ESI+).

[0914] Example 92: Preparation of 4-(8-(((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazol[1,2-b]pyridazin-6-yl)piperazine-1-carboxamide

[0915]

[0916] Step 1: 4-(8-((2-ethoxy-2-oxoethyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazolium Preparation of [1,2-b]pyridazine-6-yl)piperazine-1-carboxylic acid benzyl ester

[0917] 4-(8-((2-ethoxy-2-oxoethyl)(4-methoxybenzyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)piperazine-1-carboxylic acid benzyl ester (1.5 g, yield 35.29%) was obtained as a yellow solid using the same method as in step 1 of Example 30, except that intermediate A11 (4.48 g, 20.37 mmol) was used and the mixture was stirred for 16 hours. MS: m / z = 627.1 (M+1, ESI+).

[0918] Step 2: N-(4-methoxybenzyl)-N-(6-(piperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridyl Preparation of azinon-8-yl)glycine ethyl ester

[0919] The product obtained in step 1 (1.5 g, 2.40 mmol) was dissolved in methanol (20 mL), and then Pd / C (200 mg) was added. The mixture was then stirred at 25 °C for 48 hours under hydrogen ignition. The reaction mixture was filtered and concentrated, and the concentrate was purified by silica gel chromatography to give N-(4-methoxybenzyl)-N-(6-(piperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)glycine ethyl ester (970 mg, yield 82.20%) as a yellow solid. MS: m / z = 493.1 (M+1, ESI+).

[0920] Step 3: N-(6-(4-carbamoylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8- Preparation of ethyl glycine (N-(4-methoxybenzyl)glycine)

[0921] The product obtained in step 2 (970 mg, 1.97 mmol) and trimethylsilyl isocyanate (295 mg, 2.56 mmol) were dissolved in THF (30 mL), and then DIEA (763 mg, 5.91 mmol, 1.04 mL) was added. The mixture was stirred at 25 °C for 16 hours. Water (100 mL) was added to the reaction mixture, and the mixture was extracted three times with EtOAc (40 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give N-(6-(4-carbamoylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine ethyl ester (450 mg, 42.73% yield) as a yellow solid. MS: m / z = 536.1(M+1, ESI+).

[0922] Steps 4 to 7: 4-(8-(((6,7-difluoro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl) Preparation of imidazo[1,2-b]pyridazine-6-yl)piperazine-1-carboxamide

[0923] The compound of Example 92 (60 mg, yield 31.09%) was obtained as a white solid using the same method as in steps 2 to 5 of Example 30. However, in step 5, the product of step 3 (450 mg, 840 μmol) and intermediate B1 (583 mg, 4.05 mmol) were dissolved in DMF (15 mL) and stirred for 16 hours, and in step 7, the reaction temperature was changed to 70 °C and stirred for 16 hours. 1 H NMR (400 MHz, DMSO-d6) δ 12.69 (s, 1H), 7.88-7.85 (m, 2H), 7.25-7.16 (m, 2H), 6.23 (s, 1H), 6.05 (s, 2H), 4.85 (d, 2H), 3.36 (s, 8H); MS: m / z = 496.0 (M+1, ESI+).

[0924] Example 93: Preparation of N-((5,6-dichlorobenzo[d]thiazolyl-2-yl)methyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0925]

[0926] Step 1: Preparation of 6,6'-dithionide dimethylbis(3,4-dichloroaniline)

[0927] 5,6-Dichlorobenzo[d]thiazol-2-amine (3.6 g, 16.43 mmol) was dissolved in water (40 mL), and then NaOH (19.7 g, 493 mmol) was added at 0 °C, followed by stirring at 130 °C for 16 hours. The reaction mixture was concentrated, methanol was added, and the mixture was then filtered and concentrated to give 6,6'-dithioalkyldiylbis(3,4-dichloroaniline) (1.9 g, yield 29.94%) as a yellow solid. MS: m / z = 386.8 (M+1, ESI+).

[0928] Step 2: N-((5,6-dichlorobenzo[d]thiazo-2-yl)methyl)-N-(4-methoxybenzyl)-6-morpholino-3- Preparation of (trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0929] The product obtained in step 1 (480 mg, 1.24 mmol) was dissolved in DMF (10 mL), and then 1,4-dithiothreitol (DTT, 959 mg, 6.22 mmol) was added, followed by stirring at 25 °C for 30 min. Subsequently, intermediate S10 (600 mg, crude) was added, and the mixture was stirred at 25 °C for 16 h. Water (100 mL) was added to the reaction mixture, and the product was extracted three times with DCM (30 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give N-((5,6-dichlorobenzo[d]thiazolyl-2-yl)methyl)-N-(4-methoxybenzyl)-6-morpholino-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (330 mg, 42.58% yield) as a yellow solid. MS: m / z = 623.1 (M+1, ESI+).

[0930] Step 3: N-((5,6-dichlorobenzo[d]thiazolyl)methyl)-6-morpholino-3-(trifluoromethyl)imidazolyl Preparation of [1,2-b]pyridazine-8-amine

[0931] The compound of Example 93 (85 mg, yield 31.91%) was obtained as a white solid by the same method as step 5 of Example 30, except that the reaction time was changed to 2 hours. 1 H NMR (400 MHz, DMSO-d6) δ 8.41(s, 1H), 8.29 (s, 1H), 8.25 (t, 1H), 7.90 (s, 1H), 6.29 (s, 1H), 5.07 (d,2H), 3.67 (t, 4H), 3.35 (t, 4H); MS: m / z = 502.9 (M+1, ESI+).

[0932] Examples 94 to 108

[0933] The compounds of Examples 94 to 108 were prepared by using intermediate B2 and intermediates from the S series corresponding to the structure of the target compound and intermediates from the A series in [Table 1], with appropriate changes to the amounts of reagents, catalysts and reaction conditions, in a manner similar to that of Example 30.

[0934] [Table 6]

[0935]

[0936]

[0937]

[0938]

[0939] Examples 109 to 113

[0940] The compounds of Examples 109 to 113 were prepared by selecting and using appropriate intermediates corresponding to the structure of the target compound and intermediate B5 from the products obtained during the process of preparing the compounds of the above examples, and by appropriately changing the amount of reagents, catalysts and reaction conditions, in a manner similar to that of Example 30.

[0941] [Table 7]

[0942]

[0943]

[0944] Example 114: Preparation of 1-(1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-6-yl)piperidin-4-yl)-2-hydroxyethyl-1-one

[0945]

[0946] Step 1: Preparation of 1-(piperidin-4-yl)ethyl-1-one hydrochloride

[0947] To tert-butyl 4-acetylpiperidin-1-carboxylate (50 g, 219.97 mmol), 4 M hydrochloric acid solution (219.97 mmol, 200 mL) was added, and the mixture was stirred at 25 °C for 16 hours. The reaction mixture was concentrated to give 1-(piperidin-4-yl)ethyl-1-one hydrochloride (70 g, crude product) as a pale yellow solid. MS: m / z = 128.1 (M+1, ESI+).

[0948] Step 2: Preparation of 4-acetylpiperidine-1-carboxylic acid benzyl ester

[0949] The product obtained in step 1 (70 g, 428 mmol) was dissolved in an aqueous solution of Na₂CO₃ (500 mL), and then CbzOSu (170.57 g, 684 mmol) was added. The mixture was then stirred at 25 °C for 16 hours. The reaction mixture was filtered and the resulting solid was dried to give 4-acetylpiperidin-1-carboxylic acid benzyl ester (55 g, yield 49.20%, as a pale yellow solid. MS: m / z = 262.0 (M+1, ESI+).

[0950] Step 3: Preparation of benzyl 4-(2-bromoacetyl)piperidine-1-carboxylate

[0951] The product obtained in step 2 (55 g, 210.47 mmol) was dissolved in methanol (500 mL), and bromine (Br2, 10.8 mL, 210.47 mmol) was slowly added at 0 °C, followed by stirring at 25 °C for 16 hours. Water (2000 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (500 mL). The organic layer was washed three times with brine (2000 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give benzyl 4-(2-bromoacetyl)piperidine-1-carboxylate (30 g, yield 41.90%) as a pale yellow solid. MS: m / z = 340.0 (M+1, ESI+).

[0952] Step 4: Preparation of benzyl 4-(2-hydroxyacetyl)piperidine-1-carboxylate

[0953] The product obtained in step 3 (30 g, 88.18 mmol) was dissolved in THF (80 mL), and then potassium hydroxide (KOH, 9.90 g, 176.36 mmol) was added. The mixture was then stirred at 25 °C for 1 hour. Water (600 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (200 mL). The organic layer was washed three times with brine (600 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give benzyl 4-(2-hydroxyacetyl)piperidine-1-carboxylate (12 g, yield 49.07%) as a pale yellow solid. MS: m / z = 278.0 (M+1, ESI+).

[0954] Step 5: Preparation of benzyl 4-(2-(hydroxymethyl)-1,3-dioxolane-2-yl)piperidine-1-carboxylate

[0955] The product obtained in step 4 (12 g, 43.32 mmol) was dissolved in toluene (150 mL), and then TsOH (225 mg, 4.33 mmol) and ethane-1,2-diol (13.42 g, 216.6 mmol) were added. The mixture was then stirred at 120 °C for 48 hours. After cooling the reaction mixture, water (800 mL) was added, and the product was extracted three times with EtOAc (200 mL). The organic layer was washed three times with brine (800 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give benzyl 4-(2-(hydroxymethyl)-1,3-dioxolane-2-yl)piperidine-1-carboxylate (8.8 g, yield 63.31%) as a pale yellow solid. MS: m / z = 321.9 (M+1, ESI+).

[0956] Step 6: 4-(2-(((tert-butyldimethylsilyl)oxy)methyl)-1,3-dioxolane-2-yl)piperidine- Preparation of 1-Benzyl Formate

[0957] The product obtained in step 5 (8.8 g, 33.72 mmol) was dissolved in DMF (100 mL), and then imidazole (4.6 g, 67.44 mmol) and TBDMSCl (8.8 g, 67.44 mmol) were added. The mixture was then stirred at 25 °C for 16 hours. Water (1000 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (300 mL). The organic extract was washed three times with brine (1000 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give benzyl 4-(2-(((tert-butyldimethylsilyl)oxy)methyl)-1,3-dioxolane-2-yl)piperidine-1-carboxylate (8.4 g, yield 57.53%) as a pale yellow solid. MS: m / z = 436.1 (M+1, ESI+).

[0958] Step 7: 4-(2-(((tert-butyldimethylsilyl)oxy)methyl)-1,3-dioxolane-2-yl)piperidine Preparation

[0959] The product obtained in step 6 (8.4 g, 19.31 mmol) was dissolved in methanol (100 mL), and then Pd / C (1.16 g, 9.66 mmol) was added. The mixture was then stirred at 25 °C for 2 hours under a H2 atmosphere. The reaction mixture was filtered and concentrated to give 4-(2-(((tert-butyldimethylsilyl)oxy)methyl)-1,3-dioxolane-2-yl)piperidine (5.26 g, 90.53% yield) as a pale yellow solid. MS: m / z = 302.1 (M+1, ESI+).

[0960] Steps 8 to 12: 1-(1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoro) Preparation of (methyl)imidazo[1,2-b]pyridazin-6-yl)piperidin-4-yl)-2-hydroxyethyl-1-one

[0961] The compound of Example 114 (70 mg, yield 30.3%) was obtained as a yellow solid using the same method as steps 1 to 5 of Example 30. However, in step 8, the product obtained in step 7 (5.26 g, 17.48 mmol) was used and the mixture was stirred for 96 hours. 1H NMR (400 MHz, DMSO-d6) δ 12.58 (s, 1H), 7.86 (s, 1H), 7.82-7.76 (m, 3H), 6.15 (s, 1H), 5.01 (t, 1H), 4.83 (d, 2H), 4.16 (d, 2H), 4.04(d, 2H), 2.85 (t, 2H), 2.78-2.72 (m, 1H), 1.74 (d, 2H), 1.45-1.35 (m, 2H); MS: m / z = 542.2 (M+1, ESI+).

[0962] Example 115: Preparation of N-(1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl)imidazol[1,2-b]pyridazin-6-yl)piperidin-4-yl)-2-hydroxyacetamide

[0963]

[0964] Steps 1 to 5: 6-(4-aminopiperidin-1-yl)-N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)- Preparation of 3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0965] 6-(4-aminopiperidin-1-yl)-N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (150 mg, yield 71.43%) was obtained as a yellow solid using the same method as steps 1 to 5 of Example 30. However, in step 1 of Example 30, intermediate A12 (4.61 g, 23.07 mmol) was used and the mixture was stirred for 16 hours; in step 2, the reaction temperature was changed to 50°C and the mixture was stirred for 2 hours; and in step 4, the reaction conditions were changed to stirring at 70°C for 4 hours. MS: m / z = 499.0 (M+1, ESI+).

[0966] Step 6: N-(1-(8-(((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)amino)-3-(trifluoromethyl) Preparation of imidazo[1,2-b]pyridazine-6-yl)piperidin-4-yl)-2-hydroxyacetamide

[0967] The product obtained in step 5 (300 mg, 595 μmol) and 2-hydroxyacetamide (46 mg, 0.26 mmol) were dissolved in DCM (10 mL), and then HOBT (176 mg, 1.3 mmol), EDCI (248 mg, 1.3 mmol), and DIEA (336 mg, 2.6 mmol, 0.45 mL) were added. The mixture was then stirred at 25 °C for 16 hours. Water (80 mL) was added to the reaction mixture, and the product was extracted three times with DCM (30 mL). The organic layer was washed three times with brine (80 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel chromatography to give the compound of Example 115 (32 mg, yield 22.22%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 12.60 (s, 1H), 7.85 (d, 1H), 7.82-7.71(m, 3H), 7.59 (d, 1H), 6.18 (s, 1H), 5.36 (t, 1H), 4.83 (d, 2H), 4.02 (d,2H), 3.88-3.80 (m, 1H), 3.77 (d, 2H), 2.88 (t, 2H), 1.70 (d, 2H), 1.52-1.42(m, 2H); MS: m / z = 557.0 (M+1, ESI+).

[0968] Example 116: Preparation of N-((1H-indol-2-yl)methyl)-6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine

[0969]

[0970] Step 1: N,N-bis(4-methoxybenzyl)-6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1, Preparation of 2-b]pyridazine-8-amine

[0971] Intermediate S7 (6 g, 12.58 mmol) was dissolved in intermediate A2 (15 mL), and the mixture was stirred at 120 °C for 16 hours. Water (250 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (50 mL). The organic layer was washed three times with brine (200 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give N,N-bis(4-methoxybenzyl)-6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (6 g, 88.22% yield). MS: m / z = 541.2 (M+1, ESI+).

[0972] Step 2: Preparation of 6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine

[0973] The product obtained in step 1 (2 g, 3.7 mmol) was dissolved in DCM (15 mL), and then TFA (5 mL) was added. The mixture was stirred at 25 °C for 16 hours. The reaction mixture was concentrated, adjusted to pH 8–9 with sodium bicarbonate solution, and extracted three times with DCM (50 mL). The organic layer was washed three times with brine (200 mL), dried over Na₂SO₄, filtered, concentrated, and then purified by silica gel chromatography to give 6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (750 mg, yield 67.57%) as a yellow solid. MS: m / z = 301.1 (M+1, ESI+).

[0974] Step 3: N-((1H-indol-2-yl)methyl)-6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo Preparation of [1,2-b]pyridazine-8-amine

[0975] The product obtained in step 2 (600 mg, 2 mmol) and intermediate B21 (580.64 mg, 4 mmol) were dissolved in THF (10 mL), and Ti(OEt)4 (1.7 g, 6 mmol) was added. The mixture was then stirred at 90 °C for 16 hours. The reaction mixture was then cooled to 0 °C, and NaBH4 (302.64 mg, 8 mmol) was added. The mixture was then stirred at 0 °C for 1 hour. Water (150 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (50 mL). The organic layer was washed three times with brine (400 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by preparative HPLC to give the compound of Example 116 (60 mg, 5.55% yield) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 10.97(s, 1H), 7.84 (s, 1H), 7.64 (t, 1H), 7.44 (d, 1H), 7.33 (d, 1H), 7.04-7.00(m, 1H), 6.96-6.92 (m, 1H), 6.36 (s, 1H), 6.15 (s, 1H), 4.71 (d, 2H), 3.37(t, 4H), 2.36 (t, 4H), 2.18 (s, 3H); MS: m / z = 430.2 (M+1, ESI+).

[0976] Examples 117 to 125

[0977] The compounds of Examples 117 to 125 were prepared by using an intermediate from the intermediate series B corresponding to the structure of the target compound and by appropriately changing the amount of reagents, catalysts and reaction conditions, in a manner similar to that of Example 116.

[0978] [Table 8]

[0979]

[0980]

[0981] Example 126: Preparation of 2-(((6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)amino)methyl)-1H-benzo[d]imidazo-7-ol

[0982]

[0983] Steps 1 and 2: N-((7-methoxy-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-6- Preparation of (4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine

[0984] N-((7-methoxy-1H-benzo[d]imidazol-2-yl)methyl)-N-(4-methoxybenzyl)-6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-amine (410 mg, 82.89% yield) was obtained as a yellow solid using intermediate B8 (718.5 mg, 5.2 mmol) in the same manner as steps 5 and 6 of Example 8. However, in step 1, the reaction time was changed to 16 hours, and in step 2, the reaction temperature was changed to 70 °C with stirring for 16 hours. MS: m / z = 581.3 (M+1, ESI+).

[0985] Step 3: 2-(((6-(4-methylpiperazin-1-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)amino Preparation of (-1H-benzo[d]imidazol-7-ol)

[0986] The product obtained in step 2 (570 mg, 0.97 mmol) was dissolved in a 30% aqueous hydrobromic acid solution (HBr, 10 mL), and the mixture was stirred at 100 °C for 16 hours. After cooling the reaction mixture to room temperature, an aqueous sodium bicarbonate solution (100 mL) was added, and the product was extracted three times with EtOAc (30 mL). The organic layer was washed three times with brine (100 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was purified by preparative HPLC to give the compound of Example 126 (65 mg, yield 20.62%) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 7.85 (s, 1H), 7.72(s, 1H), 6.92 (t, 2H), 6.51 (t, 1H), 6.18 (s, 1H), 4.77 (s, 2H), 3.36 (t,4H), 2.35 (t, 4H), 2.17 (s, 3H); MS: m / z = 447.3 (M+1, ESI+).

[0987] Examples 127 to 129

[0988] The compounds of Examples 127 to 129 were prepared by using intermediate B from [Table 3], which corresponds to the structure of the target compound, and by appropriately varying the amounts of reagents, catalysts and reaction conditions, in a manner similar to that of Example 121.

[0989] [Table 9]

[0990]

[0991] Example 130: Preparation of 6-(5-aminopyridin-2-yl)-N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0992]

[0993] Step 1: Preparation of 6-(tributyltinyl)pyridine-3-amine

[0994] 6-Bromopyridin-3-amine (2.2 g, 12.72 mmol) and 1,1,1,2,2,2-hexabutyldistannane (25.83 g, 44.52 mmol) were dissolved in 1,4-dioxane (40 mL), and then P(Cy)3Pd G3 (827 mg, 1.27 mmol) was added. The mixture was stirred at 120 °C for 48 hours under a nitrogen atmosphere. Water (400 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (100 mL). The organic layer was washed three times with brine (400 mL), dried over Na2SO4, filtered, concentrated, and then purified by silica gel chromatography to give 6-(tributyltinyl)pyridin-3-amine (3.2 g, 65.68% yield) as a yellow solid. MS: m / z = 384.9 (M+1, ESI+).

[0995] Step 2: N-(6-(5-aminopyridin-2-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)-N- Preparation of (4-methoxybenzyl)glycine ethyl ester

[0996] The product obtained in step 1 (3.2 g, 8.36 mmol) and intermediate S8 (1.48 g, 3.34 mmol) were dissolved in 1,4-dioxane (20 mL), and then Pd(PPh3)4 (386 mg, 0.33 mmol) was added. The mixture was then stirred at 120 °C for 72 hours under a nitrogen atmosphere. Water (400 mL) was added to the reaction mixture, and the product was extracted three times with EtOAc (100 mL). The organic layer was washed three times with brine (400 mL), dried over Na2SO4, filtered, and concentrated. The concentrate was then purified by silica gel chromatography to give N-(6-(5-aminopyridin-2-yl)-3-(trifluoromethyl)imidazo[1,2-b]pyridazin-8-yl)-N-(4-methoxybenzyl)glycine ethyl ester (800 mg, 70.78% yield) as a yellow solid. MS: m / z = 501.3 (M+1, ESI+).

[0997] Steps 3 to 6: 6-(5-aminopyridin-2-yl)-N-((5,6-dichloro-1H-benzo[d]imidazol-2-yl)methyl)- Preparation of 3-(trifluoromethyl)imidazo[1,2-b]pyridazine-8-amine

[0998] Using the product from step 2 (300 mg, 0.6 mmol), the compound of Example 130 (33 mg, yield 18.65%) was obtained as a white solid in the same manner as steps 2 to 5 of Example 30. However, in step 3, the reaction time was changed to 16 hours; in step 4, the reaction solvent was changed to DCM and stirred for 16 hours; and in step 5, the reaction temperature was changed to 70°C and stirred for 16 hours. 1H NMR (400 MHz, DMSO-d6) δ 12.66 (s, 1H), 8.20 (t, 1H), 8.10 (s, 1H), 7.95 (d, 1H), 7.90 (d, 1H), 7.76 (s, 2H), 7.11 (s, 1H), 7.03 (dd,1H), 5.82 (s, 2H), 4.92 (d, 2H); MS: m / z = 493.1 (M+1, ESI+).

[0999] Examples 131 to 133

[1000] The compounds of Examples 131 to 133 were prepared in a manner similar to that of Example 130 by using appropriate starting materials corresponding to the structure of the target compound instead of 6-bromopyridin-3-amine, and by appropriately changing the amounts of reagents, catalysts and reaction conditions.

[1001] [Table 10]

[1002]

[1003] Experimental Example 1: Assay of CDK12 and CDK13 kinase inhibitory activity

[1004] Before initiating the CDK12 and CDK13 kinase reactions, the compounds of each example were dissolved at room temperature and diluted in DMSO as a solvent in nine concentration ranges. The diluted solutions were then incubated for 20 minutes with a mixture of kinases and substrates (and, if necessary, cofactors). Next, radiolabeled ATP was added. 33 P-γ-ATP was used to initiate the kinase reaction at room temperature for 120 minutes. After the reaction was complete, the reaction mixture was transferred to a radiolabeled enzyme catalytic product (P-γ-ATP). 33 The P-substrate was placed on filter paper. Unreacted P-substrate was removed by washing with 0.75% phosphoric acid. 33 P-ATP was then used to measure the remaining radiophosphorylated substrate on the filter paper. Kinase activity data were expressed as the percentage of remaining kinase activity in the test sample compared to the DMSO control group. The half-maximal inhibitory concentration (IC50) for each kinase was calculated using Prism (Graphpad) software. 50 ).

[1005] The CDK12 and CDK13 kinase inhibitory activities of the example compounds measured by the above methods were evaluated according to the following criteria, and the results are presented in Table 11.

[1006] A: IC 50 ≤ 0.1 μM

[1007] B: 0.1 μM < IC 50 ≤ 0.5 μM

[1008] C:IC 50 > 0.5 μM

[1009] [Table 11]

[1010]

[1011]

[1012]

[1013] As shown in Table 11 above, compared with SR-4835, the compound examples of the present invention exhibit superior CDK12 and CDK13 kinase inhibitory activity.

[1014] Experimental Example 2: Assay of CDK12 Inhibitory Activity in Cells

[1015] The cellular CDK12 inhibition of the compounds of the examples was confirmed by analyzing the Ser2 phosphorylation level in the C-terminal domain (CTD) of RNA polymerase II subunit B1 using MDA-MB-231 breast cancer cells. MDA-MB-231 cells were dispensed into 96-well plates at 100 μL of cell suspension per well using L-15 (Invitrogen) medium containing 10% FBS and incubated overnight at 37°C and 0% CO2. The next day, each of the compounds of the examples dissolved in DMSO at nine concentration ranges, along with a DMSO control group, was used to treat MDA-MB-231 cells for 6 hours at 37°C and 0% CO2. After compound treatment, the medium was removed from the 96-well plates, and 60 μL of fixation solution (4.0% paraformaldehyde in PBS) was added to each well, followed by incubation for 20 minutes. After removing the fixative and washing with PBS, 60 μL of PBS containing 0.1% Triton X-100 was dispensed into each well, followed by shaking at room temperature for 15 minutes. After washing with PBS, 100 μL of blocking buffer was added, followed by shaking at room temperature for 1 hour. After removing the blocking buffer, 60 μL of primary antibody was added to each well as the primary antibody for phosphorylated Ser2 (1:500, Cell Signaling Technology) and the C-terminal domain of RNA polymerase II subunit B1 (1:200, Cell Signaling Technology), and the reaction was incubated overnight at 4°C. After washing at least twice with 150 μL of PBS containing 0.1% Triton X-100 and shaking for 5 minutes at room temperature, 60 μL of secondary antibody was added, followed by shaking at room temperature for 1 hour in the dark. After adding 150 μL of PBS containing 0.1% Triton X-100 and washing at least twice with shaking for 5 minutes at room temperature, another 150 μL of PBS was added, followed by washing at least twice with shaking for 5 minutes at room temperature. The plate was scanned using a Li-COROdyssey instrument, with each well filled with PBS. The IC50 was calculated by fitting a dose-response curve using a 4-parameter model in Prism (Graphpad) software. 50 value.

[1016] The cellular CDK12 inhibitory activity of the compound in the examples, measured by the above method, was evaluated according to the following criteria, and the results are presented in Table 12.

[1017] IC 50 value range

[1018] A: IC 50 ≤ 0.3 μM

[1019] B: 0.3 μM < IC 50 ≤ 3 μM

[1020] C:IC 50 > 3 μM

[1021] [Table 12]

[1022]

[1023]

[1024] As shown in Table 12 above, compared with SR-4835, the compound of the present invention exhibits superior cellular CDK12 inhibitory activity.

[1025] Experimental Example 3: Determination of inhibitory activity against the proliferation of MDA-MB-231 breast cancer cells

[1026] Experiments were performed using the MDA-MB-231 breast cancer cell line to measure the cell proliferation inhibitory capacity of each compound in each example. MDA-MB-231 cells were dispensed into 96-well plates at 100 μL per well using L-15 (Ingenieur) medium containing 10% FBS, and then incubated in a cell culture incubator at 37°C and 0% CO2. The next day, cells were treated with each compound in DMSO diluted in nine concentration ranges, along with a DMSO control group, for 72 hours. After compound treatment, 100 µL of CellTiter Glo (Promega) reagent was added to each well of the 96-well plate, followed by shaking for 10 minutes and incubation at room temperature for 10 minutes. The luminescence signal was measured after inserting the 96-well plate into an Envision microplate reader (PerkinElmer). The relative IC50 was calculated by fitting dose-response curves using XLfit (IDBS LLC) software. 50 value.

[1027] The breast cancer cell proliferation inhibition activity of the compounds in the examples measured by the above methods was evaluated according to the following criteria, and the results are presented in Table 13.

[1028] IC 50 value range

[1029] A: IC 50 ≤ 0.05 µM

[1030] B: 0.05 μM < IC 50 ≤ 0.5 μM

[1031] C:IC50 > 0.5 μM

[1032] [Table 13]

[1033]

[1034]

[1035] As shown in Table 13 above, compared with SR-4835, the compound of the present invention exhibits superior inhibitory effect on MDA-MB-231 breast cancer cells.

[1036] Experimental Example 4: Assay of CDK1 and CDK2 kinase inhibitory activity and selectivity analysis relative to CDK12 kinase

[1037] Before initiating the CDK1 and CDK2 kinase reactions, the compounds of each example were dissolved at room temperature and diluted in DMSO as a solvent in nine concentration ranges. The diluted solutions were then incubated for 20 minutes with a mixture of kinases and substrates (and, if necessary, cofactors). Next, radiolabeled ATP was added. 33 P-γ-ATP was used to initiate the kinase reaction at room temperature for 120 minutes. After the reaction was complete, the reaction mixture was transferred to a radiolabeled enzyme catalytic product (P-γ-ATP). 33 The P-substrate was placed on filter paper. Unreacted P-substrate was removed by washing with 0.75% phosphoric acid. 33 P-ATP was then used to measure the remaining radiophosphorylated substrate on the filter paper. Kinase activity data were expressed as the percentage of remaining kinase activity in the test sample compared to the DMSO control group. The half-maximal inhibitory concentration (IC50) for each kinase was calculated using Prism software. 50 ).

[1038] The CDK1 and CDK2 kinase inhibitory activities of the example compounds measured by the above methods were evaluated according to the following criteria, and the results are presented in Table 14.

[1039] IC 50 value range

[1040] A: IC 50 ≤ 0.1 μM

[1041] B: 0.1 μM < IC 50 ≤ 1 μM

[1042] C:IC 50 > 1 μM

[1043] The selectivity analysis criterion relative to CDK12 is calculated as CDK1 IC. 50 With CDK12 IC50 The ratio or CDK2IC 50 With CDK12 IC 50 The ratio was evaluated according to the following criteria, and the results are summarized in Table 14 below.

[1044] A: CDK1 / CDK12 or CDK2 / CDK12 > 500x

[1045] B: 100x < CDK1 / CDK12 or CDK2 / CDK12 ≤ 500x

[1046] C: CDK1 / CDK12 or CDK2 / CDK12 ≤ 100x

[1047] [Table 14]

[1048]

[1049]

[1050] As shown in Table 14 above, the compounds of the embodiments of the present invention exhibit excellent selectivity relative to CDK12.

[1051] Experimental Example 5: Determination of Inhibitory Activity on Breast Cancer Cell Proliferation

[1052] Experiments were conducted using three types of breast cancer cell lines to measure the cell proliferation inhibitory effect of each compound in each example. For HCC70 cells, 100 µL of cell suspension was dispensed into 96-well plates using RPMI 1640 (Ingenium) medium containing 10% FBS, followed by incubation at 37°C and 5% CO2. For BT-20 cells, 100 µL of cell suspension was dispensed into 96-well plates using EMEM (Ingenium) medium containing 10% FBS, followed by incubation at 37°C and 5% CO2. For MDA-MB-436 cells, 100 µL of cell suspension was dispensed into 96-well plates using L-15 (Ingenium) medium containing 10% FBS, followed by incubation at 37°C and 0% CO2. The following day, cells were treated with each compound in each example dissolved and diluted in DMSO at nine concentration ranges, along with a DMSO control group, for 72 hours. After compound treatment, 100 µL of CellTiter Glo (Promega) reagent was added to each well of a 96-well plate, followed by shaking for 10 minutes and incubation at room temperature for 10 minutes. The luminescence signal was measured after inserting the 96-well plate into an Envision microplate reader (PerkinElmer). The relative IC50 was calculated by fitting dose-response curves using XLfit (IDBS LLC) software.50 value.

[1053] The breast cancer cell proliferation inhibitory activity of the compound in the examples, measured by the above method, was evaluated according to the following criteria, and the results are presented in Table 15.

[1054] IC 50 value range

[1055] A: IC 50 ≤ 0.05 µM

[1056] B: 0.05 µM < IC 50 ≤ 1 µM

[1057] C:IC 50 > 1 μM

[1058] [Table 15]

[1059]

[1060] As shown in Table 15 above, Examples 8, 29 and 30 of the present invention exhibited superior inhibitory effects on breast cancer cell proliferation compared with the comparative compounds palbociclib, olaparib and SR-4835.

[1061] Experimental Example 6: Determination of Inhibitory Activity Against Gastric Cancer Cell Proliferation

[1062] Experiments were conducted using four types of gastric cancer cell lines (AGS, SNU-1, KATO-III, and MKN45) to measure the cell proliferation inhibitory effect of the compounds in each embodiment. For AGS cells, 90 µL of cell suspension was dispensed into 96-well plates using F-12K (Gibco) medium containing 10% FBS, followed by incubation at 37°C and 5% CO2. For SNU-1 and MKN45 cells, 90 µL of cell suspension was dispensed into 96-well plates using RPMI 1640 (Gibco) medium containing 10% FBS, followed by incubation at 37°C and 5% CO2. For KATO-III cells, 90 µL of cell suspension was dispensed into 96-well plates using IMDM (Gibco) medium containing 10% FBS, followed by incubation at 37°C and 5% CO2. The following day, each example compound dissolved and diluted in DMSO at nine concentration ranges, along with the DMSO control group, was added in 10 µL to each 96-well plate and treated for 72 hours. After compound treatment, 100 µL of CellTiter Glo (Promega) reagent was added to each well of the 96-well plate, followed by shaking for 10 minutes and incubation at room temperature for 10 minutes. The luminescence signal was measured after inserting the 96-well plate into an Envision microplate reader (PerkinElmer). The relative IC50 was calculated by fitting dose-response curves using XLfit (IDBS LLC) software. 50 value.

[1063] The inhibitory activity of the compound in the examples, measured by the above method, on gastric cancer cell proliferation was evaluated according to the following criteria, and the results are presented in Table 16.

[1064] IC 50 value range

[1065] A: IC 50 ≤ 0.2 µM

[1066] B: 0.2 μM < IC 50 ≤ 1 μM

[1067] C:IC 50 > 1 μM

[1068] [Table 16]

[1069]

[1070] As shown in Table 16 above, compounds 8, 29 and 30 of the present invention exhibit superior inhibitory effects on gastric cancer cell proliferation compared with cisplatin, which is used as a comparative compound.

[1071] Experimental Example 7: Analysis of Cyclin K Degradation Capacity

[1072] To confirm the cyclin K degradation ability of the compounds in the examples, HCC70 breast cancer cells and MKN45 gastric cancer cells were cultured in RPMI-1640 medium supplemented with 10% FBS, and then the cells were collected to prepare cell suspensions. The HCC70 breast cancer cell suspension was divided into 2.0 × 10⁶ cells per well in 6-well plates. 6 Cells were randomly assigned and cultured in a 5% CO2 incubator at 37°C for 24 hours. MKN45 gastric cancer cell suspensions were then cultured in 6-well plates at 5.0 × 10⁶ cells per well. 6 Cells were randomly numbered and cultured in a 5% CO2 cell culture incubator at 37°C for 24 hours. The next day, each example compound dissolved and diluted in DMSO at 4 to 5 concentrations, along with a negative control group (DMSO-treated group), was added to 6 wells. HCC70 breast cancer cells were treated for 8 hours, while MKN45 gastric cancer cells were treated with the compounds for either 8 hours or 24 hours. After compound treatment, the culture medium was removed, cells were washed with PBS, and 100 µL of RIPA buffer for cell lysis was added to each of the 6 wells to collect cell lysates. The protein concentration of the cell lysates was quantified by BSA, and loading buffer was added to adjust the protein concentration to 30 µg. The mixture was then incubated at 85°C for 10 minutes, centrifuged, and allowed to stand at room temperature. After electrophoresis on a 4–12% Bis-Tris gel, the proteins were transferred to a nitrocellulose membrane. Primary antibodies, namely cyclin K (1:1000, Bethyl Lab) and focal adhesion protein (1:1000, Sigma), were attached to the membrane, followed by labeling with secondary antibody (1:10000, LI-COR). Signals were detected using an Odyssey imager (LI-COR Biosciences), and the results are presented. Figures 1 to 5 middle.

[1073] exist Figures 1 to 3 In comparison with the negative control group, Examples 8, 30, and 40 showed strong degradation of cyclin K in HCC70 cells, and... Figure 4 and 5 In comparison with the negative control group, Examples 8 and 40 showed strong degradation of cyclin K in MKN45 cells.

[1074] Experimental Example 8: Zebrafish Xenotransplantation Models of Breast and Gastric Cancer

[1075] Transgenic Tg(fli1:EGFP)y1 zebrafish embryos were cultured in E3 embryo culture medium containing 0.2 mM 1-phenyl-2-thiourea (PTU) at 28°C for 48 hours. HCC70 breast cancer cells were cultured in RPMI-1640 medium supplemented with 10% FBS, and AGS gastric cancer cells were cultured in F-12K medium supplemented with 10% FBS. Each type of cell was collected, labeled with Dil red fluorescent dye, and then subcutaneously transplanted into 2-day-old zebrafish embryos. After selecting embryos with tumors, they were classified into experimental groups (20 embryos per group) and each example compound, DMSO control group, or control compound was added, dissolved and diluted in DMSO at 2 to 3 concentration ranges. The embryos were then incubated at 35.5°C for 48 or 72 hours. In a breast cancer model, primary tumors were collected immediately after transplantation (day 0) and 48 hours later (day 2), and in a gastric cancer model, primary tumors were obtained immediately after transplantation (day 0) and 72 hours later (day 3). Images of the collected tumors were analyzed using image processing software to compare tumor size reduction with the DMSO control group, and the number of disseminated cancer cells was manually counted 3 days later using the captured images. The results are shown in [Table data would be inserted here]. Figure 6 , 7 In the breast cancer model, statistical analysis was performed on each example compound treatment group compared with the solvent group using one-way ANOVA and Dunnett's multiple comparison method (*** p < 0.001). In the gastric cancer model, statistical analysis was performed on each example compound treatment group compared with the solvent group using the Kruskal-Wallis test and Dunn's multiple comparison method (* p < 0.05). In the gastric cancer model, statistical analysis was performed on each example compound treatment group compared with the 5-FU group as a control compound using the Mann-Whitney test (* p < 0.05).

[1076] exist Figure 6 In this study, compared to the solvent, the compounds of Example 30 exhibited a dose-dependent reduction in tumor size in the HCC70 breast cancer model. Figure 7 In this study, compared to the solvent, the compound of Example 8 exhibited a dose-dependent reduction in tumor size in the AGS gastric cancer model. Figure 8 In the AGS gastric cancer model, treatment with the compound of Example 8 at 0.1 µM showed a significant reduction in the number of metastatic gastric cancer cells compared to 1 mM 5-FU as a control compound.

[1077] Experimental Example 9: Determination of Lung Cancer Cell Proliferation Inhibitory Activity

[1078] Experiments were conducted using three types of lung cancer cell lines to measure the cell proliferation inhibitory capacity of the compounds of the present invention. NCI-H23 and NCI-H358 cells were dispensed into 96-well plates at 100 µL per well using RPMI 1640 medium containing 10% FBS and 1% PS, and then incubated in a cell culture incubator at 37°C and 5% CO2. A427 cells were dispensed into 96-well plates at 100 µL per well using DMEM medium containing 10% FBS and 1% PS, and then incubated in a cell culture incubator at 37°C and 5% CO2. The following day, cells were treated for 72 hours with each example compound (e.g., Example 8) dissolved and diluted in DMSO at 10 concentration ranges, as well as a DMSO control group. After compound treatment, 10 µL of Cell Counting Kit-8 (Dojindo) reagent was added to each well of the 96-well plate, followed by incubation for 2 hours. The luminescence signal was measured after the 96-well plate was inserted into the Envision microplate reader (Thermo Fisher Scientific). The relative IC50 was calculated by fitting dose-response curves using Graphpad PRISM software. 50 value.

[1079] The lung cancer cell proliferation inhibitory activity of the compound used in the examples, measured by the above method, was evaluated according to the following criteria, and the results are presented in Table 17.

[1080] IC 50 value range

[1081] A: IC 50 ≤ 0.2 µM

[1082] B: 0.2 μM < IC 50 ≤ 1 μM

[1083] C:IC 50 > 1 μM

[1084] [Table 17]

[1085]

[1086] As shown in Table 17 above, the compound of Example 8 exhibited excellent inhibitory effects on lung cancer cell proliferation.

[1087] Experimental Example 10: Determination of Pancreatic Cancer Cell Proliferation Inhibitory Activity

[1088] Experiments were conducted using three types of pancreatic cancer cell lines to measure the cell proliferation inhibitory effect of the compounds of this invention. AsPC-1, MIA-PaCa2-control, and gemcitabine-resistant MIA-PaCa2 cells were dispensed into 96-well plates at 100 µL per well using DMEM medium containing 10% FBS and 1% PS, and then incubated in a cell culture incubator at 37°C and 5% CO2. The next day, cells were treated with the compounds of Example 8 dissolved and diluted in DMSO at 10 concentration ranges, as well as the DMSO control group, for 72 hours. After compound treatment, 10 µL of Cell Counting Kit-8 (Dongjin Chemical Research Institute) reagent was added to each well of the 96-well plate, followed by incubation for 2 hours. The luminescence signal was measured after the 96-well plates were inserted into an Envision microplate reader (Thermo Fisher Scientific). The relative IC50 was calculated by fitting dose-response curves using Graphpad PRISM software. 50 value.

[1089] The inhibitory activity of the compound in the examples, measured by the above method, on the proliferation of pancreatic cancer cells was evaluated according to the following criteria, and the results are presented in Table 18.

[1090] IC 50 value range

[1091] A: IC 50 ≤ 0.2 µM

[1092] B: 0.2 μM < IC 50 ≤ 1 μM

[1093] C:IC 50 > 1 μM

[1094] [Table 18]

[1095]

[1096] As shown in Table 18 above, the compounds in Example 8 exhibited excellent inhibitory effects on the proliferation of pancreatic cancer cells.

[1097] Experimental Example 11: Determination of Inhibitory Activity on Colorectal Cancer Cell Proliferation

[1098] Seven types of colorectal cancer cell lines were used in experiments to measure the cell proliferation inhibitory capacity of each compound in each example. DLD-1, LoVo, SW620, SW480, HCT116, HCT15, and HT29 cells were partitioned into 96-well plates at 100 µL per well using RPMI 1640 medium containing 10% FBS and 1% PS, and then incubated in a cell culture incubator at 37°C and 5% CO2. The next day, cells were treated with each compound in DMSO dissolved and diluted in 10 concentration ranges, along with a DMSO control group, for 72 hours. After compound treatment, 10 µL of Cell Counting Kit-8 (Dongjin Chemical Research Institute) reagent was added to each well of the 96-well plate, followed by incubation for 2 hours. The luminescence signal was measured after the 96-well plates were inserted into an Envision microplate reader (Thermo Fisher Scientific). The relative IC50 was calculated by fitting dose-response curves using Graphpad PRISM software. 50 value.

[1099] The inhibitory activity of the compound in the examples, measured by the above method, on the proliferation of colorectal cancer cells was evaluated according to the following criteria, and the results are presented in Table 19.

[1100] IC 50 value range

[1101] A: IC 50 ≤ 0.2 µM

[1102] B: 0.2 μM < IC 50 ≤ 1 μM

[1103] C:IC 50 > 1 μM

[1104] [Table 19]

[1105]

[1106] As shown in Table 19 above, the compounds of Examples 8 and 40 exhibited excellent inhibitory effects on the proliferation of colorectal cancer cells.

[1107] Experimental Example 12: Determination of Inhibitory Activity Against Gastric Cancer Cell Proliferation

[1108] Six types of gastric cancer cell lines were used in experiments to measure the cell proliferation inhibitory effect of the compounds in each example. NCI-N87, SNU601, KATO-III, MKN1, and AGS cells were dispensed into 96-well plates at 100 µL per well using RPMI 1640 medium containing 10% FBS and 1% PS, and then incubated in a cell culture incubator at 37°C and 5% CO2. HS746T cells were dispensed into 96-well plates at 100 µL per well using DMEM medium containing 10% FBS and 1% PS, and then incubated in a cell culture incubator at 37°C and 5% CO2. The following day, cells were treated with each example compound dissolved and diluted in DMSO at 10 concentration ranges, along with a DMSO control group, for 72 hours. After compound treatment, 10 µL of Cell Counting Kit-8 (Dongjin Chemical Research Institute) reagent was added to each well of the 96-well plate, followed by incubation for 2 hours. The luminescence signal was measured after the 96-well plate was inserted into the Envision microplate reader (Thermo Fisher Scientific). The relative IC50 was calculated by fitting dose-response curves using Graphpad PRISM software. 50 value.

[1109] The inhibitory activity of the compound in the examples, measured by the above method, on gastric cancer cell proliferation was evaluated according to the following criteria, and the results are presented in Table 20.

[1110] IC 50 value range

[1111] A: IC 50 ≤ 0.2 µM

[1112] B: 0.2 μM < IC 50 ≤ 1 μM

[1113] C:IC 50 > 1 μM

[1114] [Table 20]

[1115]

[1116] As shown in Table 20 above, the compounds of Examples 8 and 40 exhibited excellent inhibitory effects on the proliferation of gastric cancer cells.

Claims

1. A compound of formula I, its stereoisomer, hydrate, solvate, or pharmaceutically acceptable salt: [Formula I] In equation I above, R 1 The radical is halogenated, hydroxylated, cyano, C1-C6 alkoxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 ynyl, C3-C6 cycloalkyl, C3-C6 cycloalkyl-C1-C6 alkyl-, C3-C6 cycloalkyl-C2-C6 alkenyl-, or C3-C6 cycloalkyl-C2-C6 ynyl-, wherein R 1 Any C1-C6 alkoxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, and C3-C6 cycloalkyl groups may optionally be substituted with a halogen, hydroxyl, or cyano group; R 2 For C6-C 10 Aryl; a 5- or 6-membered heteroaryl group containing 1 to 2 nitrogen atoms; or a 4- to 12-membered heterocyclic group containing 1 nitrogen atom and linked via said nitrogen atom to an imidazopyridazine ring of Formula I, wherein said heterocyclic group may optionally contain an additional nitrogen atom or an oxygen atom, and R 2 It may optionally be substituted by one or more substituents selected from the group consisting of: (i) H, halogen, hydroxyl, cyano, C1-C6 alkyl, C3-C6 cycloalkyl, (ii) C1-C6 alkyl groups substituted with H, a halogen group, a hydroxyl group, or a cyano group, (iii) C3-C6 cycloalkyl groups substituted with halogen, hydroxyl, or cyano groups, (iv) amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, (C3-C6 cycloalkyl)amino, (C1-C6 hydroxyalkyl)amino, (C1-C6 alkyl)carbonylamino, (C1-C6 hydroxyalkyl)carbonylamino, or (C1-C6 alkoxy)(C1-C6 alkyl)carbonylamino, and (v) (C1-C6 alkyl)carbonyl, (C1-C6 hydroxyalkyl)carbonyl, aminocarbonyl, (C1-C6 alkyl)aminocarbonyl, di(C1-C6 alkyl)aminocarbonyl, carboxyl, or (C1-C6 alkoxy)carbonyl; and R 3 for , or ; X 1a For NR 3a O or S, and X 2a For N or CR 3a The premise is that if X 2a For CR 3a Then X 1a For NR 3a ; X 1b and X 2b One of them is N, and the other is CR. 3b ; X 2c For N or CR 3c ; X 3a X 4a X 5a and X 6a No more than one of them is N, and the rest are CR. 3d ; X 3b X 4b X 5b and X 6b No more than one of them is N, and the rest are CR. 3e ; X 3c X 4c X 5c and X 6c No more than one of them is NR 3f The rest are CR 3f R 3g ; R 3a R 3b and R 3c Each is independently H or C1-C6 alkyl; and R 3d R 3e R 3f and R 3g Each is independently H, halogen, cyano, hydroxyl, amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, C1-C6 alkoxy, or C1-C6 alkyl, wherein R 3d R 3e R 3f and R 3g Any C1-C6 alkoxy and C1-C6 alkyl groups may optionally be substituted with halogen, cyano, or hydroxyl groups.

2. The compound of formula I according to claim 1, its stereoisomers, hydrates, solvates, or pharmaceutically acceptable salts, wherein R 1 It is a halogenated group, optionally substituted with a halogenated group, a C1-C6 alkyl group, a C2-C6 alkenyl group, a C2-C6 alkynyl group, a C3-C6 cycloalkyl group, a C3-C6 cycloalkyl-C1-C6 alkyl- group, a C3-C6 cycloalkyl-C2-C6 alkenyl- group, or a C3-C6 cycloalkyl-C2-C6 alkynyl- group.

3. The compound of formula I according to claim 1, its stereoisomers, hydrates, solvates, or pharmaceutically acceptable salts, wherein R 1 It can be acetylenol, methylacetylenol, cyclopropylacetylenol, methyl, ethyl, isopropyl, cyclopropyl, trifluoromethyl, trifluoroethyl, iodo, or chloro.

4. The compound of formula I according to claim 1, its stereoisomers, hydrates, solvates, or pharmaceutically acceptable salts, wherein R 2 It is a phenyl group; pyrimidinyl, pyridinyl, pyrrolidinyl, or imidazolyl; a cyclohexacyclic butyl, pyrrolidinyl, morpholinyl, piperidinyl, or piperazine group, wherein any two non-adjacent carbon atoms of the morpholinyl, piperidinyl, or piperazine group may optionally be connected to each other by a C1-C3 alkylene group to form a bridged ring; or a 7- to 11-membered cyclohexacyclic ring containing one nitrogen atom and optionally containing one additional nitrogen atom or one oxygen atom.

5. The compound of formula I according to claim 4, its stereoisomers, hydrates, solvates, or pharmaceutically acceptable salts, wherein... The 7- to 11-membered azaspirocyclic rings are 2-azaspiro[3.3]heptane-2-yl, 2,6-diazaspiro[3.3]heptane-2-yl, 6-oxa-2-azaspiro[3.3]heptane-2-yl, 2-azaspiro[3.4]octane-2-yl, 2,6-diazaspiro[3.4]octane-2-yl, and 6-oxa-2-azaspiro[3.4]octane. -2-yl, 6-azaspiro[3.4]octane-6-yl, 2,6-diazaspiro[3.4]octane-6-yl, 2-oxa-6-azaspiro[3.4]octane-6-yl, 2-azaspiro[4.4]nonane-2-yl, 2,7-diazaspiro[4.4]nonane-2-yl, or 2-oxa-7-azaspiro[4.4]nonane-7-yl, and The bridged ring is 3-oxa-8-azabicyclo[3.2.1]octane-8-yl, 8-oxa-3-azabicyclo[3.2.1]octane-3-yl, 3,8-diazabicyclo[3.2.1]octane-3-yl, 3,8-diazabicyclo[3.2.1]octane-8-yl, 2-oxa-5-azabicyclo[2.2.2]octane-5-yl, 8-azabicyclo[3.2.1]octane-8-yl, 3-azabicyclo[3.1.1]heptane-3-yl, 6-oxa-3-azabicyclo[3.1.1]heptane-3-yl, 3,6-diazabicyclo[3.1.1]heptane-3-yl, or 3,6-diazabicyclo[3.1.1]heptane-6-yl.

6. The compound of formula I according to claim 1, its stereoisomers, hydrates, solvates, or pharmaceutically acceptable salts, wherein... R 2 For C6-C 10 Aryl or 5- or 6-membered heteroaryl containing 1 to 2 nitrogen atoms, wherein the aryl and heteroaryl are optionally selected from the group consisting of R. 2d Substitution: H, hydroxyl, amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, and (C3-C6 cycloalkyl)amino; or R 2 Choose from either formula A or formula B: [Formula A] In equation A above, n1 and n2 are each independently 1 or 2, and the carbon atoms of the ring may optionally be substituted with C1-C6 alkyl groups. If both n1 and n2 are 2, then Y 1 For CR 2a R 2b NR 2c Or O, and any two non-adjacent carbon atoms of the ring may optionally be connected to each other through a C1-C3 alkylene group to form a bridged ring, and If one or both of n1 and n2 are 1, then Y 1 For CR 2a R 2b ; [Formula B] In equation B above, n3, n4, n5, and n6 are each independently 1 or 2, and the carbon atoms of the ring can optionally be substituted with C1-C6 alkyl groups. Y 2 For CR 2a R 2b NR 2c Or O, In equations A and B above, R 2a and R 2b Each is independently selected from the group consisting of: H, halogen, hydroxyl, cyano, C1-C6 alkyl or C3-C6 cycloalkyl; C1-C6 alkyl substituted with halogen, hydroxyl or cyano; C3-C6 cycloalkyl substituted with halogen, hydroxyl or cyano; amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, (C3-C6 cycloalkyl)amino, (C1-C6 alkyl)carbonylamino, (C1-C6 hydroxyalkyl)carbonylamino; and (C1-C6 alkyl)carbonyl, (C1-C6 hydroxyalkyl)carbonyl, aminocarbonyl, (C1-C6 alkyl)aminocarbonyl, di(C1-C6 alkyl)aminocarbonyl and carboxyl; and R 2c It can be H, C1-C6 alkyl, C3-C6 cycloalkyl, (C1-C6 alkyl)carbonyl, (C1-C6 hydroxyalkyl)carbonyl, aminocarbonyl, (C1-C6 alkyl)aminocarbonyl, or di(C1-C6 alkyl)aminocarbonyl.

7. The compound of formula I according to claim 6, its stereoisomers, hydrates, solvates, or pharmaceutically acceptable salts, wherein... R 2a and R 2b One of them is selected from the group consisting of: H, halogen, hydroxyl, cyano, C1-C6 alkyl or C3-C6 cycloalkyl; C1-C6 alkyl substituted with halogen, hydroxyl or cyano; C3-C6 cycloalkyl substituted with halogen, hydroxyl or cyano; amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, (C3-C6 cycloalkyl)amino, (C1-C6 alkyl)carbonylamino, (C1-C6 hydroxyalkyl)carbonylamino; and (C1-C6 alkyl)carbonyl, (C1-C6 hydroxyalkyl)carbonyl, aminocarbonyl, (C1-C6 alkyl)aminocarbonyl, di(C1-C6 alkyl)aminocarbonyl and carboxyl, and R 2a and R 2b The other one is H, a halogen group, a hydroxyl group, a cyano group, or a C1-C6 alkyl group.

8. The compound of formula I according to claim 6, its stereoisomers, hydrates, solvates, or pharmaceutically acceptable salts, wherein... R 2a and R 2b Choose from the group consisting of: H, hydroxyl, amino, cyclopropylamino, hydroxyacetamido, methyl, hydroxymethyl, 2-hydroxyisopropyl, hydroxycyclopropyl, carboxyl, carbamoyl, hydroxyacetyl, and cyano. R 2c Choose from the group consisting of: H, methyl, cyclopropyl, acetyl, hydroxyacetyl, and carbamoyl. R 2d Choose from the following groups: hydroxyl and amino.

9. The compound of formula I according to claim 6, its stereoisomers, hydrates, solvates, or pharmaceutically acceptable salts, wherein R 2 Choose from the following groups: (In the above structure, R) 2a R 2b R 2c and R 2d Same as described in claim 6.

10. The compound of formula I according to claim 1, its stereoisomers, hydrates, solvates, or pharmaceutically acceptable salts, wherein R 2 Choose from the following groups: 。 11. The compound of formula I according to claim 1, its stereoisomers, hydrates, solvates, or pharmaceutically acceptable salts, wherein R 3 Choose from the following groups: (In the above structure, X) 3a X 4a X 5a and X 6a No more than one of them is N, and the rest are CR. 3d ;X 3b X 4b X 5b and X 6b No more than one of them is N, and the rest are CR. 3e And X 3c X 4c X 5c and X 6c No more than one of them is NR 3f The rest are CR 3f R 3g ,and R 3a R 3b and R 3c Each is independently H or C1-C6 alkyl; and R 3d R 3e R 3f and R 3g Each is independently H, halogen, cyano, hydroxyl, amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, C1-C6 alkoxy, or C1-C6 alkyl, wherein R 3d R 3e R 3f and R 3g Any C1-C6 alkoxy and C1-C6 alkyl groups may optionally be substituted with a halogen, cyano, or hydroxyl group.

12. The compound of formula I according to claim 11, its stereoisomers, hydrates, solvates, or pharmaceutically acceptable salts, wherein R 3 Choose from the following groups: (In the above structure, R) 3a R 3b and R 3c Each is independently H or C1-C6 alkyl; and R 3d R 3e R 3f and R 3g Each of these groups can be independently H, a halogen group, a hydroxyl group, an amino group, a C1-C6 alkoxy group, a C1-C6 alkyl group, or a C1-C6 haloalkyl group.

13. The compound of formula I according to claim 12, its stereoisomers, hydrates, solvates, or pharmaceutically acceptable salts, wherein R 3d R 3e R 3f and R 3g Each of the following groups is selected independently: H, F, Cl, hydroxyl, amino, methoxy, methyl, and trifluoromethyl.

14. The compound of formula I according to claim 1, its stereoisomers, hydrates, solvates, or pharmaceutically acceptable salts, wherein R 3 Choose from the following groups: 。 15. The compound of formula I according to claim 1, its stereoisomer, hydrate, solvate, or pharmaceutically acceptable salt, wherein the compound is represented by formula IA: [Form IA] In the above formula IA, R 1 The radical is halogenated, hydroxylated, cyano, C1-C6 alkoxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 ynyl, C3-C6 cycloalkyl, C3-C6 cycloalkyl-C1-C6 alkyl-, C3-C6 cycloalkyl-C2-C6 alkenyl-, or C3-C6 cycloalkyl-C2-C6 ynyl-, wherein R 1 Any C1-C6 alkoxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, and C3-C6 cycloalkyl groups may optionally be substituted with a halogen, hydroxyl, or cyano group; and Ring A is a 4- to 12-membered heterocyclic group that may optionally contain an additional nitrogen atom or an oxygen atom, and is a monocyclic, bridged, or spirocyclic ring; and Ring A may optionally be substituted by one or more substituents selected from the group consisting of: (i) H, halogen, hydroxyl, cyano, C1-C6 alkyl, C3-C6 cycloalkyl, (ii) C1-C6 alkyl groups substituted with halogen, hydroxyl, or cyano groups, (iii) C3-C6 cycloalkyl groups substituted with halogen, hydroxyl, or cyano groups, (iv) amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, (C3-C6 cycloalkyl)amino, (C1-C6 hydroxyalkyl)amino, (C1-C6 alkyl)carbonylamino, (C1-C6 hydroxyalkyl)carbonylamino, or (C1-C6 alkoxy)(C1-C6 alkyl)carbonylamino, and (v) (C1-C6 alkyl)carbonyl, (C1-C6 hydroxyalkyl)carbonyl, aminocarbonyl, (C1-C6 alkyl)aminocarbonyl, di(C1-C6 alkyl)aminocarbonyl, carboxyl or (C1-C6 alkoxy)carbonyl; R 3d It is H, halogen, cyano, hydroxyl, amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, C1-C6 alkoxy, or C1-C6 alkyl, wherein R 3d Any C1-C6 alkoxy and C1-C6 alkyl groups may optionally be substituted with halogen, cyano, or hydroxyl groups; and p is an integer from 0 to 3.

16. The compound of formula I according to claim 1, its stereoisomer, hydrate, solvate, or pharmaceutically acceptable salt, wherein the compound is represented by formula IB: [Form IB] In the above formula, R 1 The radical is halogenated, hydroxylated, cyano, C1-C6 alkoxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 ynyl, C3-C6 cycloalkyl, C3-C6 cycloalkyl-C1-C6 alkyl-, C3-C6 cycloalkyl-C2-C6 alkenyl-, or C3-C6 cycloalkyl-C2-C6 ynyl-, wherein R 1 Any C1-C6 alkoxy, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, and C3-C6 cycloalkyl groups may optionally be substituted with a halogen, hydroxyl, or cyano group; Ring B is selected from ; R 2d It can be H, hydroxyl, amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, or (C3-C6 cycloalkyl)amino; R 3d It is H, halogen, cyano, hydroxyl, C1-C6 alkoxy or C1-C6 alkyl, wherein R 3d Any C1-C6 alkoxy and C1-C6 alkyl groups may optionally be substituted with halogen, cyano, or hydroxyl groups; and p is an integer from 0 to 3.

17. The compound of formula I according to claim 16, its stereoisomers, hydrates, solvates, or pharmaceutically acceptable salts, wherein... for ,and R 2d It is an amino, (C1-C6 alkyl)amino, or di(C1-C6 alkyl)amino.

18. The compound of formula I according to claim 1, its stereoisomer, hydrate, solvate, or pharmaceutically acceptable salt, wherein the compound is represented by formula IC-1 or IC-2: In the above equation IC-1, n1 and n2 are each independently 1 or 2, and contain Y. 1 The carbon atoms of the N ring can optionally be replaced by C1-C6 alkyl groups, and If both n1 and n2 are 2, then Y 1 For CR 2a R 2b NR 2c Or O, and If one or both of n1 and n2 are 1, then Y 1 For CR 2a R 2b ; In the above equation IC-2, Y 2 For CR 2a R 2b NR 2c Or O; In the above equations IC-1 and IC-2, R 1 It can be a halogenated group, hydroxyl group, cyano group, C1-C6 alkoxy group, C1-C6 alkyl group, or C1-C6 haloalkyl group. R 3d Each is independently H, a halogen group, a hydroxyl group, a cyano group, a C1-C6 alkoxy group, a C1-C6 alkyl group, or a C1-C6 haloalkyl group, and p is an integer from 0 to 2; R 2a and R 2b Each is independently selected from the group consisting of: H, halogen, hydroxyl, cyano, C1-C6 alkyl or C3-C6 cycloalkyl; C1-C6 alkyl substituted with halogen, hydroxyl or cyano; C3-C6 cycloalkyl substituted with halogen, hydroxyl or cyano; amino, (C1-C6 alkyl)amino, di(C1-C6 alkyl)amino, (C3-C6 cycloalkyl)amino, (C1-C6 alkyl)carbonylamino, (C1-C6 hydroxyalkyl)carbonylamino; and (C1-C6 alkyl)carbonyl, (C1-C6 hydroxyalkyl)carbonyl, aminocarbonyl, (C1-C6 alkyl)aminocarbonyl, di(C1-C6 alkyl)aminocarbonyl and carboxyl; and R 2c It can be H, C1-C6 alkyl, C3-C6 cycloalkyl, (C1-C6 alkyl)carbonyl, (C1-C6 hydroxyalkyl)carbonyl, aminocarbonyl, (C1-C6 alkyl)aminocarbonyl or di(C1-C6 alkyl)aminocarbonyl.

19. A compound of formula I according to claim 1, its stereoisomer, hydrate, solvate, or pharmaceutically acceptable salt, wherein the compound is selected from the group consisting of:

20. A pharmaceutical composition comprising, as an active ingredient, a compound, stereoisomer thereof, hydrate thereof, solvate thereof or pharmaceutically acceptable salt thereof, according to any one of claims 1 to 19.

21. The pharmaceutical composition of claim 20, wherein the pharmaceutical composition is used to treat diseases caused by overexpression, excessive activity, mutation and / or activation of cyclin K-related signaling pathways of cyclin-dependent protein kinase (CDK) 12 and / or CDK 13.

22. The pharmaceutical composition according to claim 21, wherein the disease is cancer, a proliferative disease, a neurodegenerative disease, an autoimmune disease, or a disease caused by abnormal intracellular protein translation function.

23. The pharmaceutical composition according to claim 22, wherein the disease is cancer.

24. The pharmaceutical composition of claim 23, wherein the cancer is selected from the group consisting of: breast cancer, ovarian cancer, colorectal cancer, lung cancer, prostate cancer, gastric cancer, liver cancer, kidney cancer, bladder cancer, pancreatic cancer, head and neck cancer, thyroid cancer, skin cancer, bile duct cancer, esophageal cancer, hematopoietic tumors of the lymphatic and bone marrow systems, tumors of the central and peripheral nervous systems, and sarcomas.

25. The pharmaceutical composition of claim 23, wherein the pharmaceutical composition is used to treat high-grade cancer or cancer that has developed resistance to previously administered therapeutic agents.

26. The pharmaceutical composition of claim 23, wherein the pharmaceutical composition is administered in combination with one or more therapeutic agents selected from the group consisting of: radiotherapy, taxane derivatives, platinum compounds, antimetabolites, anti-CTLA4 therapy, anti-PD1 therapy, anti-PD-L1 therapy, anti-VEGF therapy, anti-EGFR therapy, topoisomerase inhibitors, anti-HER2 therapy, anti-hormonal therapy, estrogen receptor inhibitors, ERK inhibitors, PARP inhibitors, mTOR inhibitors, CDK4 / 6 inhibitors, EGFR inhibitors, HER2 inhibitors, ALK inhibitors, tyrosine kinase inhibitors, MEK inhibitors, BCR-ABL inhibitors, PI3K inhibitors, FGFR inhibitors, ROS1 inhibitors, androgen biosynthesis inhibitors, androgen receptor inhibitors, Hedgehog inhibitors, MET inhibitors, AXL inhibitors, NTRK1 inhibitors, RET inhibitors, KRAS inhibitors, and RAF inhibitors.

27. The pharmaceutical composition according to claim 22, wherein... The neurodegenerative diseases mentioned are Alzheimer's disease, Parkinson's disease, or amyotrophic lateral sclerosis (ALS); and The autoimmune diseases mentioned are rheumatoid arthritis, systemic lupus erythematosus, psoriasis, or Sjögren's syndrome.

28. A method for treating a disease caused by overexpression, hyperactivity, mutation and / or activation of cyclin K-related signaling pathways of CDK12 and / or CDK13, comprising administering to a subject a compound, stereoisomer thereof, hydrate, solvate or pharmaceutically acceptable salt thereof, according to any one of claims 1 to 19.

29. A method for inhibiting CDK12 and / or CDK13 or degrading cyclin K, comprising contacting an effective amount of the compound, stereoisomer, hydrate, solvate or pharmaceutically acceptable salt of any one of claims 1 to 19, with the cyclin-dependent kinase.

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