Imidazolo-pyridine derivatives and uses thereof

Abstract

The present invention relates to a 8-halo-imidazo[1,2-a]pyridin-7-yl-thio derivative compound, which is a low molecular weight compound that regulates SHP2 protein tyrosine phosphatase, and a method for preparing the same. A composition comprising the 8-halo-imidazo[1,2-a]pyridin-7-yl-thio small molecule compound as an active ingredient can be advantageously employed as a composition for the prevention or treatment of cancer.

Description

Imidazolo-pyridine derivatives and uses thereof

[0001] The present invention relates to an anticancer pharmaceutical composition containing an 8-halogen-imidazo[1,2-a]pyridin-7-yl-thio low molecular weight compound as an active ingredient and a method for producing the same.

[0002] SHP2 (Scr homology 2 domian-containing phosphatase 2) is a non-receptor protein tyrosine phosphatase that acts as a positive regulator of growth factor- and cytokine-mediated signaling pathways and is involved in cell proliferation, differentiation, migration, and apoptosis. SHP2 is involved in signal transduction through the RAS-mitogen-activated protein kinase (MAPK), JAK-STAT pathway, and / or phosphoinositol 3-kinase-AKT pathway (X. Yuanet et al., 2020).

[0003] SHP2 has two N-terminal Src homology 2 domains (N-SH2 and C-SH2), a catalytic domain (PTP), and a C-terminal tail. The two SH2 domains regulate the subcellular distribution and function of SHP2 (MD Asmamawet et al., 2022). In the basal state, SHP2 maintains an autoinhibitory state through the interaction of the N-SH2 domain and the PTP domain, thereby suppressing SHP2 activation. Stimulation by growth factors or cytokines acting through RTKs disrupts the binding network of the N-SH2 domain and the PTP domain, exposing the catalytic site, resulting in the enzymatic activation of SHP2 (YT Chouet et al., 2022).

[0004] Mutations in PTPN11, which encodes SHP2, disrupt the autoinhibitory state between the N-SH2 domain and the catalytic site, leading to SHP2 activation and KRAS-GTP activation, which induces RAS-associated diseases. SHP2 mutations have been found in various human diseases, including Noonan syndrome and LEOPARD syndrome, as well as juvenile myelomonocytic leukemia, acute myeloid leukemia, neuroblastoma, neuroblastoma, melanoma, breast cancer, lung cancer, hepatocellular carcinoma, and colon cancer (Yihui Song et al., 2022). In cancer cells, SHP2 is known to be associated with the RAS-ERK, JAK-STAT, PI3K-AKT, NF-kB, and mTOR signaling pathways (L. Donget et al., 2021).

[0005] KRAS is a protein involved in cell signaling pathways such as cell proliferation, maturation, and apoptosis, and when KRAS mutations occur, it induces cancer cell proliferation. KRAS mutations have been reported in 25% of all cancers, and it is estimated that KRAS mutations occur in 88% of pancreatic cancers, 50% of colon cancers, 32% of lung cancers, 20% of biliary tract cancers, and 20% of melanomas, with mutations mainly occurring in codon 12. KRAS mutations are known to activate the RAS-RAF-MEK-ERK downstream signaling pathway required for tumor cell growth by locking RAS into an active GTP-bound state, thereby activating cancer cell growth, survival, migration, metabolism, and compliance (K. Chenet et al., 2021).

[0006] SHP2 functions as a scaffold protein, and when activated, it binds to GRB2-SOS1, converting RAS-GDP to RAS-GTP, thereby activating the RAF-MEK-ERK pathway. SHP2 inhibition inhibits RAS activation by interfering with RAS-GTP binding. Notably, KARS mutant tumors are dependent on SHP2 (X. Yuanet et al., 2020).

[0007] SHP2 is a downstream regulator of the PD-1 signaling pathway, regulating cytokine production and immune responses. Enhanced antitumor immune responses have been reported in SHP2-deficient mouse models of colon cancer, and SHP2 inhibition is known to exhibit anticancer efficacy by enhancing CD8+ cytotoxic T cell-mediated antitumor immunity (M. Liuet et al., 2021).

[0008] Although various RAS mutants or MEK inhibitors have been developed, inhibition of these targets results in a negative feedback loop, leading to reactivation of upstream targets after a certain period of time, resulting in drug resistance. SHP2 activation has been reported to play a key role in this process.

[0009] Therefore, SHP2, which is activated or up-regulated in human cancers and other diseases, may be a good therapeutic target, and the compound of the present invention that can inhibit SHP2 activity has high potential as a novel small molecule therapeutic agent for treating various diseases.

[0010] Meanwhile, international patent publication WO 2023-169170 A1 and international patent publication WO 2023-046198 A1 provide methods for preparing and using compounds as SHP2 inhibitors that are potentially useful for treating SHP2-related diseases.

[0011] [Chemical Formula I] of International Publication No. WO 2023-046198 is characterized by the inclusion of sulfaketone, and thus differs from the chemical structure of the present invention. In addition, International Publication No. WO 2023-169170 A1 is a compound in which the 2nd position of the imidazo[1,2-a]pyridin-7-yl substituent is substituted with hydrogen or a methyl group, and the present invention is a compound in which the 2nd position of the imidazo[1,2-a]pyridin-7-yl substituent is substituted with an amide-containing substituent, heteroaryl, or heterocycloalkyl, and thus differs in chemical structure.

[0012] The present inventors have compared the compounds developed in the existing literature with the imidazo[1,2-a]pyridin-7-yl substituent (R2) in the chemical formula 1 of the present invention. , , The present invention was completed by confirming that a compound substituted with an aryl group having a 6-10 member ring, a heteroaryl group having a 5-10 member ring, or a heterocycloalkyl group having a 4-10 member ring exhibits excellent physiological activity against cancer cells.

[0013] The purpose of the present invention is to provide an anticancer pharmaceutical composition containing an 8-halogen-imidazo[1,2-a]pyridin-7-yl-thio low molecular weight compound as an active ingredient and a method for producing the same.

[0014] The present invention relates to a compound comprising a low molecular weight 8-halogen-imidazo[1,2-a]pyridin-7-yl-thio compound represented by the following formula 1, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof.

[0015] [Chemical Formula 1]

[0016]

[0017] In the above formula,

[0018] R1 is hydrogen, C1-C6alkoxyC1-C6alkylNHC(=O)-, , or and;

[0019] R2 is C1-C6alkoxy, C1-C6alkoxyC1-C6alkyl, C1-C6alkylC(=O)NHC1-C6alkyl-, C1-C6alkoxyC1-C6alkylNHC(=O)-, , , or And,

[0020] In the above formula, A is a substituted or unsubstituted 6-10 membered aryl, a substituted or unsubstituted 5-10 membered heteroaryl, or a substituted or unsubstituted 4-10 membered heterocycloalkyl;

[0021] R3 is halogen;

[0022] R4 is hydrogen or C1-C6 alkyl;

[0023] R5 is substituted or unsubstituted 2-oxa-8-azaspiro[4.5]decan-8-yl or substituted or unsubstituted 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl,

[0024] wherein the substituted 2-oxa-8-azaspiro[4.5]decan-8-yl or 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl is each independently substituted with one or more substituents selected from the group consisting of hydrogen, amino, and C1-C6 alkyl;

[0025] R6 is hydroxymethyl, C1-C6alkoxymethyl or C1-C6alkoxymethoxymethyl;

[0026] R7 is each independently hydrogen, halogen, amino, cyano, acetyl, carboxyl, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkoxy-C(=O)-, C1-C6 alkyl-C(=O)-, C1-C6 alkyl-C(=O)NH-, C1-C6 alkylNH-, C1-C6 alkyl-NHC(=O)-, C1-C6 alkyl-OC(=O)NH-, C1-C6 alkyl-SO2NH-, C1-C6 alkoxy-C1-C6 alkyl-NHC(=O)-(C1-C6 alkoxy) 0-1 -, C1-C6 alkyl-NHC(=O)-(C1-C6 alkoxy) 0-1 -, COOH-(C1-C6alkoxy) 0-1 -, , , or and;

[0027] m is 0, 1, or 2;

[0028] The present invention relates to a compound of the following formula 2, which is composed of an 8-halogen-imidazo[1,2-a]pyridin-7-yl-thio low molecular weight compound, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof.

[0029] [Chemical Formula 2]

[0030]

[0031] In the above formula,

[0032] R1 is hydrogen;

[0033] R2 is C1-C6 alkylC(=O)NHC1-C6 alkyl-, C1-C6 alkoxyC1-C6 alkylNHC(=O)-, , , or And,

[0034] In the above formula, if A is phenyl or pyridinyl, R7 is each independently hydrogen, halogen, amino, cyano, acetyl, carboxyl, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkoxy-C(=O)-, C1-C6 alkyl-C(=O)-, C1-C6 alkyl-C(=O)NH-, C1-C6 alkylNH-, C1-C6 alkyl-NHC(=O)-, C1-C6 alkyl-OC(=O)NH-, C1-C6 alkyl-SO2NH-, C1-C6 alkoxy-C1-C6 alkyl-NHC(=O)-(C1-C6 alkoxy) 0-1 -, C1-C6 alkyl-NHC(=O)-(C1-C6 alkoxy) 0-1 -, COOH-(C1-C6alkoxy) 0-1 -, , , or and;

[0035] In the above formula, if A is pyrrolyl, furanyl, thiophenyl or pyrimidinyl, R7 is each independently hydrogen, halogen, amino, cyano, acetyl, carboxyl, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkoxy-C(=O)-, C1-C6 alkyl-C(=O)-, C1-C6 alkyl-C(=O)NH-, C1-C6 alkylNH-, C1-C6 alkyl-NHC(=O)-, C1-C6 alkyl-OC(=O)NH-, C1-C6 alkyl-SO2NH-, C1-C6 alkoxy-C1-C6 alkyl-NHC(=O)-(C1-C6 alkoxy) 0-1-, C1-C6 alkyl-NHC(=O)-(C1-C6 alkoxy) 0-1 -, or COOH-(C1-C6alkoxy) 0-1 - and;

[0036] In the above formula, if A is piperidinyl, R7 is each independently hydrogen, halogen, amino, cyano, acetyl, carboxyl, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkoxy-C(=O)-, C1-C6 alkyl-C(=O)-, C1-C6 alkyl-C(=O)NH-, C1-C6 alkylNH-, C1-C6 alkyl-NHC(=O)-, C1-C6 alkyl-OC(=O)NH-, C1-C6 alkyl-SO2NH-, C1-C6 alkoxy-C1-C6 alkyl-NHC(=O)-(C1-C6 alkoxy) 0-1 -, C1-C6 alkyl-NHC(=O)-(C1-C6 alkoxy) 0-1 -, or COOH-(C1-C6alkoxy) 0-1 - and;

[0037] R3 is halogen;

[0038] R4 is hydrogen or C1-C6 alkyl;

[0039] R5 is substituted or unsubstituted 2-oxa-8-azaspiro[4.5]decan-8-yl or substituted or unsubstituted 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl,

[0040] wherein the substituted 2-oxa-8-azaspiro[4.5]decan-8-yl or 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl is each independently substituted with one or more substituents selected from the group consisting of hydrogen, amino, and C1-C6 alkyl;

[0041] R6 is hydroxymethyl, C1-C6alkoxymethyl or C1-C6alkoxymethoxymethyl;

[0042] m is 0, 1, or 2;

[0043] The present invention relates to a compound of the following formula 2, which is composed of an 8-halogen-imidazo[1,2-a]pyridin-7-yl-thio low molecular weight compound, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof.

[0044] In the above chemical formula 2,

[0045] R1 is hydrogen;

[0046] R2 is And,

[0047] In the above formula, if A is phenyl or pyridinyl, R7 is each independently hydrogen, halogen, amino, cyano, acetyl, carboxyl, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkoxy-C(=O)-, C1-C6 alkyl-C(=O)-, C1-C6 alkyl-C(=O)NH-, C1-C6 alkylNH-, C1-C6 alkyl-NHC(=O)-, C1-C6 alkyl-OC(=O)NH-, C1-C6 alkyl-SO2NH-, C1-C6 alkoxy-C1-C6 alkyl-NHC(=O)-(C1-C6 alkoxy) 0-1 -, C1-C6 alkyl-NHC(=O)-(C1-C6 alkoxy) 0-1 -, COOH-(C1-C6alkoxy) 0-1 -, , , or and;

[0048] In the above formula, if A is pyrrolyl, furanyl, thiophenyl or pyrimidinyl, R7 is each independently hydrogen, halogen, amino, cyano, acetyl, carboxyl, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkoxy-C(=O)-, C1-C6 alkyl-C(=O)-, C1-C6 alkyl-C(=O)NH-, C1-C6 alkylNH-, C1-C6 alkyl-NHC(=O)-, C1-C6 alkyl-OC(=O)NH-, C1-C6 alkyl-SO2NH-, C1-C6 alkoxy-C1-C6 alkyl-NHC(=O)-(C1-C6 alkoxy) 0-1 -, C1-C6 alkyl-NHC(=O)-(C1-C6 alkoxy) 0-1-, or COOH-(C1-C6alkoxy) 0-1 - and;

[0049] In the above formula, if A is piperidinyl, R7 is each independently hydrogen, halogen, amino, cyano, acetyl, carboxyl, C1-C6 alkyl, haloC1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkoxy-C(=O)-, C1-C6 alkyl-C(=O)-, C1-C6 alkyl-C(=O)NH-, C1-C6 alkylNH-, C1-C6 alkyl-NHC(=O)-, C1-C6 alkyl-OC(=O)NH-, C1-C6 alkyl-SO2NH-, C1-C6 alkoxy-C1-C6 alkyl-NHC(=O)-(C1-C6 alkoxy) 0-1 -, C1-C6 alkyl-NHC(=O)-(C1-C6 alkoxy) 0-1 -, or COOH-(C1-C6alkoxy) 0-1 - and;

[0050] R3 is halogen;

[0051] R4 is hydrogen or C1-C6 alkyl;

[0052] R5 is substituted or unsubstituted 2-oxa-8-azaspiro[4.5]decan-8-yl or substituted or unsubstituted 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl,

[0053] wherein the substituted 2-oxa-8-azaspiro[4.5]decan-8-yl or 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl is each independently substituted with one or more substituents selected from the group consisting of hydrogen, amino, and C1-C6 alkyl;

[0054] R6 is hydroxymethyl, C1-C6alkoxymethyl or C1-C6alkoxymethoxymethyl;

[0055] m is 0, 1, or 2;

[0056] The present invention relates to a compound of formula 2, which is composed of an 8-halogen-imidazo[1,2-a]pyridin-7-yl-thio low molecular weight compound, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof.

[0057] The present invention, in the above chemical formula 2,

[0058] In the above formula,

[0059] R5 is substituted or unsubstituted 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl,

[0060] Here, the above-mentioned substituted 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl is independently substituted with one or more substituents selected from the group consisting of hydrogen, amino, and C1-C6 alkyl.

[0061] The present invention relates to a compound of formula 2, which is composed of an 8-halogen-imidazo[1,2-a]pyridin-7-yl-thio low molecular weight compound, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof.

[0062] The present invention, in the above chemical formula 2,

[0063] R3 is chloro; R4 is hydrogen or methyl;

[0064] In addition, the present invention relates to a compound represented by the chemical formula 1, characterized in that it is one selected from the group of compounds below, or an enantiomer, diastereomer or stereoisomer thereof, a hydrate, a solvate, a prodrug or a pharmaceutically acceptable salt thereof.

[0065] (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(4-methoxyphenyl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 2);

[0066] (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(methoxymethyl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 4);

[0067] N-((7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)methyl)acetamide hydrochloride (Compound 5);

[0068] (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(pyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 6);

[0069] (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(1H-pyrrol-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 7);

[0070] (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(thiophen-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 8);

[0071] (7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-2-methylimidazo[1,2-a]pyridin-3-yl)(piperazin-1-yl)methanone dihydrochloride (Compound 9);

[0072] (7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-2-methylimidazo[1,2-a]pyridin-3-yl)(4-aminopiperidin-1-yl)methanone dihydrochloride (Compound 10);

[0073] 7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-N-(2-methoxyethyl)-2-methylimidazo[1,2-a]pyridine-3-carboxamide hydrochloride (Compound 11);

[0074] 7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-2-methyl-N-(piperidin-4-yl)imidazo[1,2-a]pyridine-3-carboxamide dihydrochloride (Compound 12);

[0075] (7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)(piperazin-1-yl)methanone dihydrochloride (Compound 13);

[0076] (7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)(4-aminopiperidin-1-yl)methanone dihydrochloride (Compound 14);

[0077] 7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-N-(piperidin-4-yl)imidazo[1,2-a]pyridine-2-carboxamide dihydrochloride (Compound 15);

[0078] 7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-N-(2-methoxyethyl)imidazo[1,2-a]pyridine-2-carboxamide hydrochloride (Compound 16);

[0079] 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(piperazin-1-yl)ethanone dihydrochloride (Compound 17);

[0080] 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(4-aminopiperidin-1-yl)ethanone dihydrochloride (Compound 18);

[0081] 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(piperidin-4-yl)acetamide dihydrochloride (Compound 19);

[0082] 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(2-methoxyethyl)acetamide hydrochloride (Compound 20);

[0083] 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-methylacetamide hydrochloride (Compound 21);

[0084] 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetic acid hydrochloride (Compound 22);

[0085] (S)-2-(3-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetic acid hydrochloride (Compound 23);

[0086] 2-(3-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(piperazin-1-yl)ethan-1-one dihydrochloride (Compound 24);

[0087] 2-(3-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(2-methoxyethyl)acetamide hydrochloride (Compound 25);

[0088] 4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)benzoic acid hydrochloride (Compound 26);

[0089] (4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenyl)(piperazin-1-yl)methanone dihydrochloride (Compound 27);

[0090] 4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-N-(2-methoxyethyl)benzamide hydrochloride (Compound 28);

[0091] (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((2-(4-aminophenyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 29);

[0092] N-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenyl)acetamide hydrochloride (Compound 30);

[0093] (S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(4-methoxyphenyl))imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 31);

[0094] (S)-7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-N-(2-methoxyethyl)imidazo[1,2-a]pyridine-2-carboxamide hydrochloride (Compound 32);

[0095] (S)-2-(4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetic acid hydrochloride (Compound 33);

[0096] (S)-2-(4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(piperazin-1-yl)ethanone dihydrochloride (Compound 34);

[0097] (S)-2-(4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(4-aminopiperidin-1-yl)ethan-1-one dihydrochloride (Compound 35);

[0098] (S)-2-(4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(piperidin-4-yl)acetamide dihydrochloride (Compound 36);

[0099] (S)-2-(4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(2-methoxyethyl)acetamide hydrochloride (Compound 37);

[0100] (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(4-methoxyphenyl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol (Compound 38);

[0101] (S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(4-methoxyphenyl))imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol (Compound 39);

[0102] (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(furan-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 40);

[0103] (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(6-chloropyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 41);

[0104] (S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(6-methoxypyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol hydrochloride (Compound 42);

[0105] tert-Butyl ((3S,4S)-8(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(piperidin-4-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol dihydrochloride (Compound 43);

[0106] (S)-1-(4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)piperidin-1-yl)ethan-1-one (Compound 44);

[0107] Ethyl (S)-4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)piperidine-1-carboxylate (Compound 45);

[0108] (S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(5-methoxypyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol hydrochloride (Compound 46);

[0109] (S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(2-methoxypyrimidin-4-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol hydrochloride (Compound 47);

[0110] (S)-4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-N-(2-methoxyethyl)benzamide (Compound 48);

[0111] (S)-4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-N-methylbenzamide (Compound 49);

[0112] (S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((2-(4-aminophenyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol (Compound 50);

[0113] (S)-N-(4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenyl)acetamide (Compound 51);

[0114] (S)-4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-2-fluoro-N-methylbenzamide hydrochloride (Compound 52);

[0115] (S)-6-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-2-chloro-N-methylnicotinamide (Compound 53);

[0116] tert-Butyl (S)-(1'-(5-((8-chloro-2-(6-(methylcarbamoyl)pyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (Compound 54); and

[0117] (S)-6-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-N-methylpicolinamide (Compound 55).

[0118] Additionally, the following terms in the present invention have the following meanings unless otherwise specified. Any undefined term has the meaning understood in the art.

[0119] The term “alkyl” above means a straight or branched chain hydrocarbon group having 1 to 20 carbon atoms and preferably 1 to 6 carbon atoms, and includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, 1-methylpropyl, pentyl and hexyl.

[0120] The above term “alkoxy” means an oxygen group bonded to a straight or branched chain saturated hydrocarbon having 1 to 20 carbon atoms, and preferably 1 to 6 carbon atoms, and includes methoxy, ethoxy, propoxy, n-butoxy, tert-butoxy, 1-methylpropoxy, etc.

[0121] The terms “halo” and “halogen” are used in their usual sense to refer to fluoro, chloro, bromo or iodo substituents.

[0122] The term "haloalkyl" above refers to an alkyl group in which one or more hydrogen atoms of the alkyl group are replaced by a halo group, wherein the alkyl and halo groups are as disclosed above.

[0123] The term "haloalkoxy" above refers to an alkoxy group in which one or more hydrogen atoms of the alkoxy group are replaced by a halo group, wherein halo and alkoxy are as disclosed above.

[0124] The above term "aryl" refers to an aromatic ring compound, including but not limited to phenyl, naphthalene, anthracene, etc.

[0125] The above term “heteroaryl” refers to an aromatic ring compound containing one or more heteroatoms such as N, O, or S, and includes, but is not limited to, pyrrolyl, thiophenyl, pyrazolyl, imidazolyl, isoxazolyl, triazolyl, pyridinyl, pyrazinyl, pyrimidinyl, furanyl, indolyl, indazolyl, benzoimidazolyl, benzoisoxazolyl, benzoxazolyl, benzothiazolyl, benzothiophenyl, naphthofuranyl, quinolinyl, isoquinolinyl, quinoxalinyl, etc., depending on the number and type of heteroatoms contained in the ring and the number of carbon atoms.

[0126] “Heterocycloalkyl group” means a cycloalkyl group as defined above containing one to three identical or different heteroatoms selected from oxygen, nitrogen and sulfur within the ring system, including but not limited to oxetanyl, azetidinyl, tetrahydrofuranyl, dioxolanyl, dithiolanyl, tetrahydropyranyl, piperidinyl, quinuclidinyl, morpholinyl, piperazinyl, dioxanyl or dithianyl.

[0127] Furthermore, the compound of formula 1 of the present invention may include not only pharmaceutically acceptable salts, but also all salts, hydrates, solvates, and prodrugs that can be prepared by conventional methods. In addition, the compound of the present invention may contain one or more asymmetric carbon atoms and may exist in racemic and optically active forms. All such compounds and diastereomers are included within the scope of the present invention.

[0128] In the present invention, the pharmaceutically acceptable salt means a salt or complex of Chemical Formula 1 having a desirable biological activity. Examples of such salts include, but are not limited to, acid addition salts formed with inorganic acids [e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, etc.], and salts formed with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, fumaric acid, maleic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalene sulfonic acid, naphthalene disulfonic acid, and poly-galacturonic acid. The compound may also be administered as a pharmaceutically acceptable quaternary salt known to those skilled in the art, in particular, chloride, bromide, iodide, -O-alkyl, toluenesulfonate, methylsulfonate, sulfonate, phosphate, or carboxylate (e.g., benzoate, succinate, acetate, glycolate, maleate, malate, fumarate, citrate, tartrate, ascorbate, cinnamoate, mandeloate, and diphenylacetate). The compound of formula 1 of the present invention may include not only pharmaceutically acceptable salts, but also all salts, hydrates, solvates, and prodrugs that can be prepared by conventional methods.

[0129] The acid addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the derivative of chemical formula 1 in an organic solvent such as methanol, ethanol, acetone, dichloromethane, acetonitrile, etc., adding an organic acid or inorganic acid, filtering and drying the resulting precipitate, or by distilling the solvent and an excess acid under reduced pressure, drying, and crystallizing in the presence of an organic solvent.

[0130] Additionally, pharmaceutically acceptable metal salts can be prepared using bases. Alkali metal or alkaline earth metal salts can be obtained, for example, by dissolving a compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering out the undissolved compound salt, and evaporating and drying the filtrate. Among the metal salts, sodium, potassium, or calcium salts are pharmaceutically suitable. Furthermore, the corresponding salts can be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

[0131] The present invention relates to a pharmaceutical composition characterized in that it prevents or treats cancer using an 8-halogen-imidazo[1,2-a]pyridin-7-yl-thio low molecular weight compound.

[0132] The above cancer means a solid cancer or blood cancer, and the above solid cancer is brain cancer, brain tumor, benign astrocytoma, malignant astrocytoma, pituitary adenoma, meningioma, brain lymphoma, oligodendroglioma, intracranial tumor, ependymoma, brainstem tumor, head and neck tumor, glioma, glioblastoma, laryngeal cancer, oropharyngeal cancer, nasal cancer, nasopharyngeal cancer, salivary gland cancer, hypopharyngeal cancer, tonsil cancer, thyroid cancer, oral cancer, esophageal cancer, eye cancer, thoracic tumor, small cell lung cancer, non-small cell lung cancer, thymic cancer, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, pleural cancer, mediastinal tumor, breast cancer, male breast cancer, abdominal tumor, stomach cancer, gastric mastoid tumor, liver cancer, hepatoblastoma, gallbladder cancer, biliary tract cancer, pancreatic cancer, small intestine cancer, colon cancer, colon cancer, rectal cancer, duodenal cancer, anal cancer, bladder cancer, It may be selected from the group consisting of kidney cancer, renal pelvis cancer, heart cancer, peritoneal cancer, adrenal cancer, spinal cord cancer, bone cancer, genital tumors, penile cancer, prostate cancer, female genital tumors, cervical cancer, endometrial cancer, ovarian cancer, uterine sarcoma, vaginal cancer, female external genital cancer, female urethral cancer, or skin cancer, but is not particularly limited thereto. In addition, the blood cancer may be selected from the group consisting of acute myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, childhood lymphoma, malignant lymphoma, multiple myeloma, or aplastic anemia, but is not particularly limited thereto.

[0133] The pharmaceutical composition according to the present invention can be formulated into a suitable form together with a pharmaceutically acceptable carrier commonly used. "Pharmaceutically acceptable" refers to a composition that is physiologically tolerable and does not typically cause allergic reactions or similar reactions such as gastrointestinal disorders or dizziness when administered to humans. In addition, the composition can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injectable solutions, each according to a conventional method.

[0134] Carriers, excipients and diluents that may be included in the above composition include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl parahydroxybenzoate, propyl parahydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, the composition is prepared using diluents or excipients such as commonly used fillers, stabilizers, binders, disintegrants and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations are prepared by mixing the compound of the present invention with at least one excipient, such as starch, microcrystalline cellulose, sucrose or lactose, low-substituted hydroxypropyl cellulose, hypromellose, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, syrups, etc., and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, flavoring agents, preservatives, etc. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases include witepsol, macrogol, Tween 61, cocoa butter, laurin, glycerol, and gelatin.In order to formulate a formulation for parenteral administration, the compound of the above chemical formula 1 or a pharmaceutically acceptable salt thereof is mixed in water together with sterilized or preservatives, stabilizers, wetting agents or emulsifying promoters, salts or buffers for osmotic pressure control, and other therapeutically useful substances to prepare a solution or suspension, which can be prepared in an ampoule or vial unit dosage form.

[0135] The pharmaceutical composition comprising the compound of chemical formula 1 disclosed in the present invention as an active ingredient can be administered to mammals such as mice, livestock, and humans by various routes. All modes of administration are conceivable, and for example, it can be administered orally, rectally, or by intravenous, intramuscular, subcutaneous, intrauterine, or intracerebrovascular injection. The dosage will vary depending on the age, sex, and body weight of the subject to be treated, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the time of administration, the route of administration, the absorption, distribution, and excretion rates of the drug, the types of other drugs used, and the judgment of the prescriber. Determination of the dosage based on these factors is within the level of those skilled in the art, and the dosage generally ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferred dosage is 1 mg / kg / day to 500 mg / kg / day. The dosage can be administered once a day or divided into several doses. The above dosage does not limit the scope of the present invention in any way.

[0136] In addition, the pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, hormone therapy, chemotherapy, and biological response modifiers for the prevention or treatment of cancer.

[0137] The present invention relates to an anticancer pharmaceutical composition containing an 8-halogen-imidazo[1,2-a]pyridin-7-yl-thio low molecular weight compound as an active ingredient and a method for producing the same. A composition containing the low molecular weight derivative as an active ingredient can be usefully used as a composition for preventing or treating cancer.

[0138] Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the content presented herein is intended to be thorough and complete, and to fully convey the spirit of the present invention to those skilled in the art.

[0139] <Example 1. Synthesis of compound and confirmation of physicochemical properties>

[0140] The synthesis process of compounds 1 to 55 of the present invention is as follows.

[0141]

[0142] Compound 1. (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-methylimidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride

[0143] Step 1) Preparation of ethyl 6-bromo-3-chloro-5-methylpyrazine-2-carboxylate (1)

[0144] To a solution of triphenylphosphine (22.61 g, 86.19 mmol, 3.00 eq) in 1,4-dioxane (300 mL) was added N-chlorosuccinimide (11.71 g, 87.63 mmol, 3.05 eq) at room temperature for 0.5 h. After this time, the solution became a white slurry. Then, ethyl 6-bromo-3-hydroxy-5-methylpyrazine-2-carboxylate (7.50 g, 28.73 mmol, 1.00 eq) was added to this slurry, and the resulting mixture was heated to 100 °C for 5 h. The reaction mass was then cooled to room temperature, triethylamine (24 mL) was added, and the mixture was concentrated. The residue was quenched with water and extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4 and the solvent was evaporated. The concentrate was purified by silica gel column chromatography using Hex / EtOAc (2 / 8) as the eluent to obtain 1 (6.56 g, 23.46 mmol, 82%) as a colorless oil.

[0145] 1 H NMR (400 MHz, DMSO-d6) δ 4.41 (q,J= 7.1 Hz, 2H), 2.65 (s, 3H), 1.34 (t,J= 7.1 Hz, 3H).

[0146] Step 2) Preparation of ethyl 6-((2-amino-3-chloropyridin-4-yl)thio)-3-chloro-5-methylpyrazine-2-carboxylate (2)

[0147] To a solution of 1 (240 mg, 0.86 mmol, 1.00 eq) in 1,4-dioxane (5.0 mL) were added 2-amino-3-chloro-pyridine-4-thio, sodium salt (188 mg, 1.03 mmol, 1.20 eq), Pd2(dba)3 (39 mg, 0.043 mmol, 0.05 eq), Xantphos (50 mg, 0.086 mmol, 0.10 eq), and DIEA (0.60 mL, 3.44 mmol, 4.00 eq) at room temperature. The mixture was purged with N2 and evacuated three times. The reaction mixture was stirred at 95 °C for 3 h. The reaction was quenched with water and extracted with EtOAc (2 x 20 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (3 / 7) as the eluent to give 2 (187 mg, 0.52 mmol, 61%) as a pale yellow solid.

[0148] ESI-MS m / z 358.8 [M+H + ]

[0149] Step 3) Preparation of ethyl 3-chloro-6-((8-chloro-2-methylimidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazine-2-carboxylate (3)

[0150] To a solution of 2 (130 mg, 0.36 mmol, 1.0 eq) in ethanol (10 mL) at room temperature was added chloroacetone (0.14 mL, 1.81 mmol, 5.0 eq). The reaction mixture was stirred at 100 °C for 16 h, cooled to room temperature, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using Hex / EtOAc (1 / 1) as the eluent to give 3 (120 mg, 0.30 mmol, 83%) as a pale yellow solid.

[0151] 1H NMR (400 MHz, DMSO-d6) δ 8.69 (d,J= 7.1 Hz, 1H), 8.07 (s, 1H), 7.28 (d,J= 7.1 Hz, 1H), 4.27 (q,J= 7.1 Hz, 2H), 2.64 (s, 3H), 2.47 (s, 3H), 1.15 (t,J= 8.9, 5.3 Hz, 3H). ESI-MS m / z 397.0 [M+H + ]

[0152] Step 4) Preparation of ethyl 3-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-methylimidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazine-2-carboxylate (4)

[0153] To a solution of 3 (120 mg, 0.30 mmol, 1.0 eq) and (3S,4S)-3-methyl-2-oxa-8-azaspiro[4,5]decan-4-amine dihydrochloride (88 mg, 0.36 mmol, 1.2 eq) in DMA (5.0 mL) was added DIEA (0.31 mL, 1.80 mmol, 6.0 eq) at room temperature. The reaction mixture was stirred at 85 °C for 4 h. The reaction was cooled to room temperature, Boc2O (262 mg, 1.20 mmol, 4.0 eq) was added to the reaction solution, and the resulting brown solution was allowed to stand at room temperature for 16 h. The reaction was quenched with brine, water, and then extracted with EtOAc (2 x 20 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (1 / 1) as the eluent to obtain 4 (130 mg, 0.21 mmol, 68%) as a pale yellow solid.

[0154] 1H NMR (500 MHz, DMSO-d6) δ 8.31 (d,J= 7.1 Hz, 1H), 7.74 (s, 1H), 7.02 (d,J= 10.6 Hz, 1H), 6.37 (s, 1H), 4.27 (q,J= 7.0 Hz, 2H), 4.19-4.13 (m, 1H), 3.90 (dd,J= 10.5, 4.9 Hz, 1H), 3.67 (d,J= 8.4 Hz, 1H), 3.58-3.40 (m, 5H), 2.42 (s, 3H), 2.34 (s, 3H), 1.72-1.63 (m,J= 25.0, 6.2 Hz, 3H), 1.54-1.48 (m, 1H), 1.41-1.39 (m, 10H), 1.24 (t,J= 7.1 Hz, 3H), 1.03 (d,J= 6.3 Hz, 3H). ESI-MS m / z 631.2 [M+H + ]

[0155] Step 5) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-methylimidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (5)

[0156] To a solution of 4 (100 mg, 0.16 mmol, 1.0 eq) in dichloromethane (DCM, 10 mL) at -78 °C was added dropwise 1 M DIBAL-H in THF (0.79 mL, 0.79 mmol, 5.0 eq). The solution was then stirred at -78 °C for 10 min and then warmed to room temperature for 1 h. The reaction was quenched by pouring into a saturated aqueous Rochelle salt solution. The resulting slurry was warmed to room temperature with vigorous stirring for 1 h, after which DCM and water were added. The organic layer was separated, and the aqueous layer was extracted with DCM (2 x 20 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (2 / 8) as the eluent to give 5 (43 mg, 0.073 mmol, 36%) as a pale yellow solid.

[0157] 1 H NMR (500 MHz, DMSO-d6) δ 8.30 (d,J= 7.1 Hz, 1H), 7.74 (s, 1H), 7.01 (d,J= 10.5 Hz, 1H), 6.36 (d,J= 7.1 Hz, 1H), 5.31 (t,J= 5.8 Hz, 1H), 4.43 (d,J= 5.5 Hz, 2H), 4.17 (dt,J= 12.6, 6.2 Hz, 1H), 3.88 (dd,J= 10.5, 4.9 Hz, 1H), 3.68 (d,J= 8.4 Hz, 1H), 3.53 (d,J= 8.5 Hz, 1H), 3.51-3.46 (m, 1H), 3.41-3.34 (m, 2H), 3.25-3.19 (m, 1H), 2.42 (s, 3H), 2.34 (s, 3H), 1.77-1.66 (m, 3H), 1.61-1.54 (m, 1H), 1.40 (s, 9H), 1.03 (d,J= 6.3 Hz, 3H). ESI-MS m / z 589.1 [M+H + ]

[0158] Step 6) Preparation of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-methylimidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (compound 1)

[0159] To a solution of 5 (15 mg, 0.025 mmol, 1.0 eq) in DCM (5.0 mL) was added 4N-HCl (0.13 mL, 0.050 mmol, 20 eq) in 1,4-dioxane at 0 °C. The resulting mixture was stirred at room temperature for 16 h. The solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 1 (6.2 mg, 0.012 mmol, 46%) as a pale yellow solid.

[0160] 1 H NMR (400 MHz, DMSO-d6) δ 8.56 (d,J= 7.2 Hz, 1H), 8.19 (s, 3H), 8.03 (s, 1H), 6.77 (d,J= 7.3 Hz, 1H), 4.47 (s, 2H), 4.26-4.18 (m, 1H), 3.97-3.84 (m, 3H), 3.68 (d,J= 9.1 Hz, 1H), 3.13-3.03 (m, 2H), 2.45 (d,J= 4.3 Hz, 6H), 2.02-1.81 (m, 4H), 1.77-1.71 (m,J= 12.8 Hz, 1H), 1.66-1.61 (m,J= 12.5 Hz, 1H), 1.24 (d,J= 6.3 Hz, 3H). ESI-MS m / z 489.1 [M+H + ]

[0161]

[0162] Compound 2. (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(4-methoxyphenyl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride

[0163] Step 1) Preparation of ethyl 6-bromo-3-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-5-methylpyrazine-2-carboxylate (6)

[0164] To a solution of 1 (5.00 g, 17.89 mmol, 1.0 eq) and (3S,4S)-3-methyl-2-oxa-8-azaspiro[4,5]decan-4-amine dihydrochloride (5.22 g, 21.47 mmol, 1.2 eq) in DMA (70 mL) was added DIEA (18.75 mL, 107.34 mmol, 6.0 eq) at room temperature. The reaction mixture was stirred at 85 °C for 4 h. The reaction was cooled to room temperature, and Boc2O (15.62 g, 71.56 mmol, 4.0 eq) was added to the reaction mixture. The resulting brown solution was allowed to stand at room temperature for 16 h. The reaction was quenched with brine, water, and then extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed under vacuum. The residue was purified by silica gel column chromatography using Hex / EtOAc (3 / 7) as the eluent to give 6 (6.45 g, 12.56 mmol, 70%) as a pale yellow solid.

[0165] 1H NMR (400 MHz, CDCl3) δ 4.46-4.38 (m, 2H), 4.25-4.15 (m, 1H), 3.81 (d,J= 8.8 Hz, 1H), 3.74-3.53 (m, 3H), 3.33 (d,J= 9.4 Hz, 1H), 3.26-3.16 (m, 1H), 3.03 (d,J= 4.6 Hz, 1H), 2.54 (d,J= 2.6 Hz, 3H), 1.90 (s, 1H), 1.82-1.61 (m, 4H), 1.41 (t,J= 7.1 Hz, 3H), 1.26 (t,J= 4.9 Hz, 3H).

[0166] Step 2) Preparation of tert-butyl ((3S,4S)-8-(5-bromo-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (7)

[0167] To a solution of 6 (6.45 g, 12.56 mmol, 1.00 eq) in DCM (100 ml) at -78 °C was added dropwise 1 M DIBAL-H in THF (62.80 ml, 62.80 mmol, 5.00 eq). The solution was then stirred at -78 °C for 0.5 h and then warmed to room temperature for 0.5 h. The reaction was quenched by pouring into a saturated aqueous Rochelle salt solution. The resulting slurry was warmed to room temperature under vigorous stirring for 1 h, after which DCM and water were added. The organic layer was separated, and the aqueous layer was extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (3 / 7) as the eluent to give 7 (3.85 g, 8.17 mmol, 65%) as a pale yellow solid.

[0168] 1H NMR (400 MHz, CDCl3) δ 4.70-4.59 (m, 3H), 4.22-4.16 (m, 1H), 3.99 (dd,J= 10.7, 4.4 Hz, 1H), 3.73-3.67 (m, 2H), 3.44-3.34 (m, 1H), 3.31-3.23 (m, 1H), 3.15-3.07 (m, 1H), 3.05-2.92 (m, 1H), 2.54 (s, 3H), 1.96-1.80 (m, 3H), 1.72-1.61 (m, 2H), 1.47 (s, 9H), 1.21 (d,J= 6.3 Hz, 3H). ESI-MS m / z 471.0 [M+H + ]

[0169] Step 3) Preparation of tert-butyl ((3S,4S)-8-(5-((2-amino-3-chloropyridin-4-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (8)

[0170] To a solution of 7 (3.85 g, 8.17 mmol, 1.00 eq) in 1,4-dioxane (30 mL) at room temperature were added 2-amino-3-chloro-pyridine-4-thio, sodium salt (2.24 g, 12.26 mmol, 1.50 eq), Pd2(dba)3 (374 mg, 0.41 mmol, 0.05 eq), Xantphos (474 ​​mg, 0.82 mmol, 0.10 eq), and DIEA (4.28 mL, 24.51 mmol, 3.00 eq). The mixture was purged with N2 and evacuated three times. The reaction mixture was stirred at 95 °C for 16 h. The reaction was quenched with water and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed under vacuum. The residue was purified by silica gel column chromatography using Hex / EtOAc (3 / 7) as the eluent to give 8 (3.02 g, 5.48 mmol, 67%) as a pale yellow solid.

[0171] 1H NMR (500 MHz, DMSO-d6) δ 7.65 (s, 1H), 7.01 (d,J= 10.5 Hz, 1H), 6.33 (d,J= 14.5 Hz, 2H), 5.75 (d,J= 5.3 Hz, 1H), 5.42 (t,J= 5.7 Hz, 1H), 4.46 (d,J= 5.7 Hz, 2H), 4.20-4.13 (m, 1H), 3.89 (dd,J= 10.5, 4.9 Hz, 1H), 3.68 (d,J= 8.3 Hz, 1H), 3.53 (d,J= 8.3 Hz, 1H), 3.46-3.40 (m, 3H), 3.31-3.25 (m, 1H), 2.40 (s, 3H), 1.79-1.63 (m,J= 14.5 Hz, 3H), 1.62-1.52 (m,J= 23.5 Hz, 1H), 1.39 (s, 9H), 1.05 (dd,J= 22.0, 6.2 Hz, 3H). ESI-MS m / z 551.0 [M+H + ]

[0172] Step 4) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-(4-methoxyphenyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (9)

[0173] To a solution of 8 (80 mg, 0.145 mmol, 1.0 eq) in MeCN (2.0 mL) was added 2-chloro-1-(4-methoxyphenyl)ethanone (27 mg, 0.145 mmol, 1.0 eq) at room temperature. The reaction mixture was stirred at 100 °C for 16 h, cooled to room temperature, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using DCM / MeOH (19 / 1) as an eluent to give 9 (18 mg, 0.026 mmol 18%) as a pale yellow solid.

[0174] 1H NMR (500 MHz, CDCl3) δ 7.94 (d,J= 6.8 Hz, 1H), 7.88 (d,J= 8.5 Hz, 2H), 7.78 (s, 1H), 6.92 (d,J= 8.6 Hz, 2H), 6.51 (d,J= 7.0 Hz, 1H), 4.57 (d,J= 10.7 Hz, 2H), 4.53 (s, 1H), 4.16-4.09 (m, 1H), 3.94 (dd,J= 10.7, 4.3 Hz, 1H), 3.89-3.73 (m, 4H), 3.68-3.62 (m, 2H), 3.45-3.37 (m, 1H), 3.37-3.27 (m, 1H), 3.21-3.10 (m, 1H), 3.09-2.99 (m, 1H), 2.50 (s, 3H) 1.90-1.73 (m, 3H), 1.64-1.57 (m, 1H), 1.48 (s, 9H), 1.16 (d,J= 6.2 Hz, 3H). ESI-MS m / z 681.4 [M+H + ]

[0175] Step 5) Preparation of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(4-methoxyphenyl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (compound 2)

[0176] To a solution of 9 (18 mg, 0.026 mmol, 1.0 eq) in DCM (3.0 mL) at 0 °C was added 4N-HCl (0.07 mL, 0.26 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 16 h. The solvent was removed under vacuum to give compound 2 (7.8 mg, 0.013 mmol, 48%) as a pale yellow solid.

[0177] 1H NMR (500 MHz, DMSO-d6) δ 8.35 (d,J= 7.7 Hz, 2H), 7.90 (d,J= 8.7 Hz, 2H), 7.03 (d,J= 8.7 Hz, 2H), 6.39 (d,J= 7.1 Hz, 1H), 5.35 (t,J= 5.8 Hz, 1H), 4.45 (d,J= 5.6 Hz, 2H), 4.12-3.99 (m, 2H), 3.81 (s, 3H), 3.67 (d,J= 8.4 Hz, 1H), 3.66-3.57 (m, 2H), 3.50 (d,J= 8.4 Hz, 1H), 3.29-3.23 (m, 1H), 3.22-3.14 (m, 1H), 2.91 (d,J= 5.0 Hz, 1H), 2.44 (s, 3H), 2.04-1.94 (m, 2H), 1.87-1.81 (m, 1H), 1.75-1.69 (m, 1H), 1.61-1.52 (m,J= 22.9 Hz, 2H), 1.09 (d,J= 6.4 Hz, 3H).

[0178]

[0179] Compound 3. (3S,4S)-8-(5-((8-chloro-2-methylimidazo[1,2-a]pyridin-7-yl)thio)-3-((methoxymethoxy)methyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-amine

[0180] Step 1) Preparation of tert-butyl ((3S,4S)-8-(5-bromo-3-((methoxymethoxy)methyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (10)

[0181] To a solution of 7 (150 mg, 0.32 mmol, 1.0 eq) in DCM (2.0 mL) at 0 °C was added DIEA (0.32 mL, 1.92 mmol, 6.0 eq). MOMCl (0.12 mL, 1.60 mmol, 5.0 eq) was added dropwise to the reaction solution, and the reaction was stirred at room temperature for 16 h. The reaction was quenched with water and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (1 / 9) as an eluent to give 10 (169 mg, 0.33 mmol, 99%) as a yellow solid.

[0182] 1 H NMR (400 MHz, DMSO-d6) δ 6.98 (d,J= 10.4 Hz, 1H), 4.69 (s, 2H), 4.49 (s, 2H), 4.14 (q,J= 5.9 Hz, 1H), 3.86 (dd,J= 10.5, 4.9 Hz, 1H), 3.66 (d,J= 8.4 Hz, 1H), 3.30 (s, 3H), 3.25-3.19 (m, 2H), 3.07 (d,J= 9.6 Hz, 1H), 2.46 (s, 3H), 1.78-1.65 (m, 3H), 1.55 (t,J= 9.3 Hz, 1H), 1.39 (s, 9H), 1.02 (d,J= 6.3 Hz, 3H).

[0183] Step 2) Preparation of tert-butyl ((3S,4S)-8-(5-((2-amino-3-chloropyridin-4-yl)thio)-3-((methoxymethoxy)methyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (11)

[0184] To a solution of 10 (160 mg, 0.310 mol, 1.0 eq) in 1,4-dioxane (5.0 mL) at room temperature were added 2-amino-3-chloro-pyridine-4-thio, sodium salt (62.4 mg, 0.341 mmol, 1.1 eq), Pd2(dba)3 (14 mg, 0.016 mmol, 0.05 eq), Xantphos (18 mg, 0.031 mmol, 0.10 eq), and DIEA (0.16 mL, 0.93 mmol, 4.00 eq). The mixture was purged with N2 and evacuated three times. The reaction mixture was stirred at 95 °C for 8 h. The reaction was quenched with water and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed under vacuum. The residue was purified by silica gel column chromatography using DCM / MeOH (19 / 1) as the eluent to give 11 (160 mg, 0.269 mmol, 86%) as a yellow solid.

[0185] 1 H NMR (400 MHz, DMSO-d6) δ 7.65 (d,J= 5.4 Hz, 1H), 7.00 (d,J= 10.6 Hz, 1H), 6.34 (s, 2H), 5.78 (d,J= 5.4 Hz, 1H), 5.75 (s, 1H), 4.66 (s, 2H), 4.53 (s, 2H), 4.22-4.11 (m, 1H), 3.89 (dd,J= 10.5, 5.0 Hz, 1H), 3.68 (d,J= 8.3 Hz, 2H), 3.27 (s, 5H), 3.18 (t,J= 5.6) Hz, 1H), 2.41 (s, 3H), 1.81-1.64 (m, 3H), 1.55 (s, 1H), 1.40 (s, 9H), 1.03 (d,J= 6.3 Hz, 3H).

[0186] Step 3) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-methylimidazo[1,2-a]pyridin-7-yl)thio)-3-((methoxymethoxy)methyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (12)

[0187] To a solution of 11 (69 mg, 0.116 mmol, 1.0 eq) in ethanol (1.0 mL) at room temperature were added chloroacetone (48 μL, 0.579 mmol, 5.0 eq) and KI (3.7 mg, 0.022 mmol, 0.2 eq). The reaction mixture was stirred at 55 °C for 16 h. The reaction mass was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using Hex / EtOAc (1 / 9) as an eluent to give 12 (22 mg, 0.347 mmol, 30%) as a yellow solid.

[0188] 1 H NMR (500 MHz, MeOD) δ 8.21-8.14 (m, 1H), 7.61 (dd,J= 3.9, 1.7 Hz, 1H), 6.94 (dd,J= 10.7, 2.5 Hz, 1H), 6.54 (ddd,J= 7.1, 3.0, 1.5 Hz, 1H), 4.64 (d,J= 1.3 Hz, 2H), 4.58-4.53 (m, 2H), 4.28-4.22 (m, 1H), 3.98 (dd,J= 10.7, 4.7 Hz, 1H), 3.75 (d,J= 8.6 Hz, 1H), 3.68 (dd,J=8.8, 2.0 Hz, 1H), 3.60-3.54 (m, 1H), 3.49-3.39 (m, 2H), 3.26 (d,J= 1.1 Hz, 4H), 2.55-2.48 (m, 3H), 2.43 (d,J= 1.8 Hz, 3H), 1.93-1.78 (m, 3H), 1.67 (ddd,J= 13.4, 7.6, 3.2 Hz, 1H), 1.48 (s, 9H), 1.17 (d,J= 6.3 Hz, 3H). ESI-MS m / z 633.1 [M+H + ]

[0189] Step 4) Preparation of (3S,4S)-8-(5-((8-chloro-2-methylimidazo[1,2-a]pyridin-7-yl)thio)-3-((methoxymethoxy)methyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-amine (compound 3)

[0190] To a solution of 12 (22 mg, 0.035 mmol, 1.0 eq) in DCM (1.0 mL) at 0 °C was added 70% TFA in DCM (1.0 mL). The resulting mixture was stirred at room temperature for 6 h. The solvent was removed in vacuo, and the residue was dissolved in methanol (1.0 mL) and TEA (2 drops). The reaction was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as the eluent to give compound 3 (3.6 mg, 0.006 mmol, 19%) as a pink oil.

[0191] 1 H NMR (500 MHz, MeOD) δ 8.19 (d,J= 7.1 Hz, 1H), 7.64 (s, 1H), 6.58 (d,J= 7.1 Hz, 1H), 4.63 (s, 2H), 4.57 (s, 2H), 4.30-4.22 (m, 1H), 3.87 (d,J= 8.7 Hz, 1H), 3.74 (d,J= 8.7 Hz, 1H), 3.72-3.64 (m, 2H), 3.27-3.18 (m, 4H), 3.13 (ddd,J= 12.8, 10.0, 2.6 Hz, 1H), 3.07 (d,J= 4.9 Hz, 1H), 2.52 (s, 3H), 2.44 (s, 3H), 1.98-1.89 (m, 2H), 1.73 (dd,J= 21.6, 14.6 Hz, 2H), 1.25 (d,J= 6.4 Hz, 3H). ESI-MS m / z 533.0 [M+H + ]

[0192]

[0193] Compound 4. (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(methoxymethyl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride

[0194] Step 1) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-(chloromethyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (13)

[0195] To a solution of 8 (290 mg, 0.53 mmol, 1.0 eq) in ethanol (10 mL) at room temperature was added 1,3-dichloro-2-propanone (81 mg, 0.64 mmol, 1.2 eq). The reaction mixture was stirred at 85 °C for 16 h. The reaction mass was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using DCM / MeOH (19 / 1) as the eluent to give 13 (51 mg, 0.08 mmol, 16%) as a yellow solid.

[0196] 1 H NMR (400 MHz, CDCl3) δ 7.92 (d,J= 7.1 Hz, 1H), 7.67 (s, 1H), 6.58 (d,J= 7.2 Hz, 1H), 4.82 (s, 2H), 4.63 (d,J= 10.6 Hz, 1H), 4.59 (s, 2H), 4.22-4.17 (m, 1H), 4.00 (dd,J= 10.7, 4.3 Hz, 1H), 3.71 (s, 2H), 3.52-3.43 (m, 1H), 3.42-3.36 (m, 1H), 3.26-3.18 (m, 1H), 3.10 (t,J=8.4 Hz, 1H), 2.56 (s, 3H), 1.97-1.78 (m, 4H), 1.48 (s, 9H), 1.22 (d,J= 6.3 Hz, 3H). ESI-MS m / z 623.2 [M+H + ]

[0197] Step 2) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-(methoxymethyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (14)

[0198] To a solution of 13 (15 mg, 0.024 mmol, 1.0 eq) in methanol (2.0 mL) at room temperature was added sodium methoxide (5.2 mg, 0.096 mmol, 4.0 eq). The reaction mixture was stirred at 60 °C for 18 h. The reaction mass was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as the eluent to give 14 (9.0 mg, 0.015 mmol, 60%) as a yellow solid.

[0199] 1 H NMR (500 MHz, CDCl3) δ 7.92 (d,J= 7.1 Hz, 1H), 7.59 (s, 1H), 6.56 (d,J= 7.1 Hz, 1H), 4.70 (s, 2H), 4.64 (d,J= 10.7 Hz, 1H), 4.57 (s, 2H), 4.21-4.15 (m, 1H), 3.99 (dd,J= 10.7, 4.3 Hz, 1H), 3.73-3.68 (m, 2H), 3.52 (s, 3H), 3.48-3.40 (m, 1H), 3.40-3.33 (m, 1H), 3.24-3.16 (m, 1H), 3.10-3.03 (m, 1H), 2.55 (s, 3H), 1.96-1.78 (m, 4H), 1.68-1.62 (m, 1H), 1.49 (d,J= 15.9 Hz, 9H), 1.22 (d,J= 6.3 Hz, 3H). ESI-MS m / z 619.7 [M+H + ]

[0200] Step 3) Preparation of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(methoxymethyl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (compound 4)

[0201] To a solution of 14 (9.0 mg, 0.015 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (40 μL, 0.150 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h. The solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 4 (6.9 mg, 0.012 mmol, 85%) as a pale yellow solid.

[0202] 1 H NMR (500 MHz, MeOD) δ 8.54 (d,J= 7.1 Hz, 1H), 8.19 (s, 1H), 6.98 (d,J= 7.1 Hz, 1H), 4.72 (s, 2H), 4.67 (s, 2H), 4.33 (dd,J= 6.4, 4.1 Hz, 1H), 4.03-3.96 (m, 2H), 3.96-3.86 (m, 2H), 3.53-3.45 (m, 4H), 3.24-3.10 (m, 2H), 2.56 (s, 3H), 2.03-1.96 (m, 2H), 1.96-1.89 (m, 1H), 1.79-1.73 (m, 1H), 1.35 (d,J= 6.5 Hz, 3H). ESI-MS m / z 519.0 [M+H + ]

[0203]

[0204] Compound 5. N-((7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)methyl)acetamide hydrochloride

[0205] Step 1) Preparation of tert-butyl ((3S,4S)-8-(5-((2-(azidomethyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (15)

[0206] To a solution of 13 (40 mg, 0.064 mmol, 1.0 eq) in DMF (1.0 mL) was added NaN3 (6.2 mg, 0.096 mmol, 1.5 eq) at room temperature. The resulting mixture was stirred at room temperature for 2 h. The reaction was quenched with water and brine, then extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo to give 15 (35 mg, 0.056 mmol) as a yellow oil, which was used without further purification.

[0207] ESI-MS m / z 630.6 [M+H + ]

[0208] Step 2) Preparation of tert-butyl ((3S,4S)-8-(5-((2-(aminomethyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (16)

[0209] To a solution of 15 (35 mg, 0.056 mmol, 1.0 eq) in MeOH (2.0 mL) was added PPh3 (22 mg, 0.084 mmol, 1.5 eq) at room temperature. The resulting mixture was stirred at 80 °C for 2 h. After removing methanol under vacuum, the residue was diluted with water (5.0 mL), and the mixture was extracted with EtOAc (2 X 25 mL). The organic layer was dried over Na2SO4, filtered, and concentrated. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 16 (9.0 mg, 0.015 mmol, 58% (2 steps)) as a yellow solid.

[0210] 1 H NMR (400 MHz, CDCl3) δ 7.92 (d,J= 7.1 Hz, 1H), 7.60 (s, 1H), 6.48 (d,J= 7.0 Hz, 1H), 4.67 (d,J= 10.8 Hz, 1H), 4.58 (s, 2H), 4.22-4.15 (m, 1H), 3.98 (dd,J= 10.7, 4.3 Hz, 1H), 3.76-3.64 (m, 2H), 3.49-3.41 (m, 1H), 3.41-3.32 (m, 1H), 3.25-3.15 (m, 1H), 3.12-3.01 (m, 1H), 2.89-2.71 (m, 5H), 2.53 (s, 3H), 1.95-1.76 (m, 3H), 1.68-1.61 (m, 1H), 1.47 (s, 9H), 1.21 (d,J= 6.2 Hz, 3H). ESI-MS m / z 604.7 [M+H + ]

[0211] Step 3) Preparation of tert-butyl ((3S,4S)-8-(5-((2-(acetamidomethyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (17)

[0212] To a solution of 16 (9.0 mg, 0.015 mmol, 1.0 eq) in DCM (2.0 mL) at 0 °C was added acetic anhydride (7.1 μL, 0.075 mmol, 5.0 eq). The resulting mixture was stirred at room temperature for 1 h. The reaction was quenched with water and extracted with DCM (2 X 20 mL). The organic layer was dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 17 (6.6 mg, 0.010 mmol, 69%) as a yellow solid.

[0213] 1 H NMR (500 MHz, CDCl3) δ 7.90 (d,J= 7.1 Hz, 1H), 7.58 (s, 1H), 6.56 (d,J= 7.1 Hz, 1H), 6.54-6.45 (m, 1H), 4.64 (d,J= 10.8 Hz, 1H), 4.60-4.57 (m, 3H), 4.23-4.16 (m, 1H), 3.99 (dd,J= 10.7, 4.3 Hz, 1H), 3.75-3.66 (m, 2H), 3.51-3.42 (m, 1H), 3.42-3.33 (m, 1H), 3.27-3.16 (m, 1H), 3.15-3.04 (m, 1H), 2.55 (s, 3H), 2.07 (s, 2H), 2.03 (s, 3H), 1.95-1.83 (m, 3H), 1.67 (d,J= 3.6 Hz, 1H), 1.49 (d,J= 15.8 Hz, 9H), 1.22 (d,J= 6.3 Hz, 3H). ESI-MS m / z 646.0 [M+H + ]

[0214] Step 4) Preparation of N-((7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)methyl)acetamide hydrochloride (compound 5)

[0215] To a solution of 17 (6.6 mg, 0.010 mmol, 1.0 eq) in DCM (3.0 mL) at 0 °C was added 4N-HCl (0.03 mL, 0.102 mmol, 10 eq) dissolved in dioxane. The resulting mixture was stirred at room temperature for 1 h. The solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring at room temperature for 5 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 5 (4.7 mg, 0.008 mmol, 81%) as a pale yellow solid.

[0216] 1 H NMR (400 MHz, MeOD) δ 8.49 (d,J= 7.0 Hz, 1H), 8.08 (s, 1H), 6.96 (d,J= 6.9 Hz, 1H), 4.66 (s, 2H), 4.62 (s, 2H), 4.38-4.28 (m, 1H), 4.06-3.83 (m, 4H), 3.49 (d,J= 3.8 Hz, 1H), 3.24-3.07 (m, 2H), 2.56 (s, 3H), 2.06 (s, 3H), 2.04-1.90 (m, 3H), 1.80-1.73 (m, 1H), 1.35 (d,J= 6.5 Hz, 3H). ESI-MS m / z 546.1 [M+H + ]

[0217]

[0218] Compound 6. (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(pyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride

[0219] Step 1) Preparation of 2-bromo-1-(pyridin-2-yl)ethenone (18)

[0220] To a solution of 2-acetylpyridine (300 mg, 2.48 mmol, 1.0 eq) in chloroform (10 mL) at room temperature were added NBS (529 mg, 2.97 mmol, 1.2 eq) and pTsOHㆍH2O (94 mg, 0.50 mmol, 0.2 eq). The reaction mixture was stirred at 70 °C for 18 h. The reaction was quenched with saturated aqueous NaHCO3, water, and extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (2 / 8) as an eluent to give 18 (100 mg, 0.50 mmol, 20%) as a white solid.

[0221] 1 H NMR (400 MHz, CDCl3) δ 8.72 (d,J= 4.7 Hz, 1H), 8.13 (d,J= 7.8 Hz, 1H), 7.89 (dd,J= 7.8, 1.7 Hz, 1H), 7.60-7.51 (m, 1H), 4.89 (s, 2H). ESI-MS m / z 199.9 [M+H + ]

[0222] Step 2) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-(pyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (19)

[0223] To a solution of 8 (30 mg, 0.054 mmol, 1.0 eq) in ethanol (2.0 mL) at room temperature were added 18 (13.0 mg, 0.065 mmol, 1.2 eq) and KI (1.8 mg, 0.011 mmol, 0.2 eq). The reaction mixture was stirred at 65 °C for 16 h. The reaction mass was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using DCM / MeOH (19 / 1) as an eluent to give 19 (19 mg, 0.029 mmol, 54%) as a white solid.

[0224] 1 H NMR (500 MHz, CDCl3) δ 8.65 (d,J= 4.7 Hz, 1H), 8.59-8.46 (m, 1H), 8.46-8.36 (m, 1H), 8.00 (d,J= 7.1 Hz, 1H), 7.92 (d,J= 7.2 Hz, 1H), 7.41-7.32 (m, 1H), 6.56 (d,J= 7.1 Hz, 1H), 4.63 (d,J= 10.7 Hz, 1H), 4.60 (s, 2H), 4.22-4.16 (m, 1H), 4.04-3.97 (m, 1H), 3.74-3.68 (m, 2H), 3.52-3.44 (m, 1H), 3.43-3.37 (m, 1H), 3.27-3.20 (m, 1H), 3.14-3.06 (m, 1H), 2.57 (s, 3H), 1.97-1.90 (m, 1H), 1.88-1.78 (m, 2H), 1.70-1.63 (m, 1H), 1.48 (s, 9H), 1.22 (d,J= 6.2 Hz, 3H). ESI-MS m / z 652.0 [M+H + ]

[0225] Step 3) Preparation of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(pyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (compound 6)

[0226] To a solution of 19 (12 mg, 0.018 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.05 mL, 0.180 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for several minutes, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 6 (10 mg, 0.017 mmol, 93%) as a pale yellow solid.

[0227] 1 H NMR (500 MHz, MeOD) δ 8.83 (s, 1H), 8.79 (d,J= 5.6 Hz, 1H), 8.52-8.36 (m, 3H), 7.89-7.81 (m, 1H), 6.76 (d,J= 7.2 Hz, 1H), 4.65 (s, 2H), 4.36-4.30 (m, 1H), 3.99 (d,J= 9.1 Hz, 1H), 3.96-3.82 (m, 3H), 3.49 (d,J= 3.9 Hz, 1H), 3.21-3.14 (m, 1H), 3.14-3.07 (m, 1H), 2.57 (s, 3H), 2.03-1.90 (m, 3H), 1.79-1.72 (m, 1H), 1.35 (d,J= 6.5 Hz, 3H). ESI-MS m / z 552.0 [M+H + ]

[0228]

[0229] Compound 7. (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(1H-pyrrol-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride

[0230] Step 1) Preparation of 2-bromo-1-(1H-pyrrol-2-yl)ethenone (20)

[0231] To a solution of 2-acetylpyrrole (300 mg, 2.75 mmol, 1.0 eq) in chloroform (10 mL) at room temperature, N-bromosuccinimide (NBS, 587 mg, 3.30 mmol, 1.2 eq) and pTsOHㆍH2O (105 mg, 0.55 mmol, 0.2 eq) were added. The reaction mixture was stirred at 70 °C for 18 h. The reaction was washed with NaHCO 3(aq) , quenched with water and extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4 and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (2 / 8) as eluent to give 20 (217 mg, 1.15 mmol, 42%) as a white solid.

[0232] 1 H NMR (500 MHz, CDCl3) δ 7.67 (d,J= 0.8 Hz, 1H), 7.37 (d,J= 3.6 Hz, 1H), 6.63 (dd,J= 3.6, 1.6 Hz, 1H), 4.35 (s, 2H). ESI-MS m / z 188.9 [M+H + ]

[0233] Step 2) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-(1H-pyrrol-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (21)

[0234] To a solution of 8 (30 mg, 0.054 mmol, 1.0 eq) in ethanol (2.0 mL) at room temperature were added 20 (12 mg, 0.065 mmol, 1.2 eq) and KI (1.8 mg, 0.011 mmol, 0.2 eq). The reaction mixture was stirred at 65 °C for 16 h and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using DCM / MeOH (19 / 1) as an eluent to give 21 (7.1 mg, 0.011 mmol, 20%) as a white solid.

[0235] 1 H NMR (500 MHz, CDCl3) δ 7.67 (d,J= 0.8 Hz, 1H), 7.37 (d,J= 3.6 Hz, 1H), 6.63 (dd,J= 3.6, 1.6 Hz, 1H), 4.35 (s, 2H). 1 H NMR (500 MHz, CDCl3) δ 8.00 (d,J= 6.4 Hz, 1H), 7.83 (s, 1H), 7.51 (s, 1H), 6.62 (d,J= 6.6 Hz, 1H), 6.54 (s, 1H), 4.66-4.59 (m, 3H), 4.23-4.17 (m, 1H), 4.01 (dd,J= 10.7, 4.2 Hz, 1H), 3.74-3.69 (m, 2H), 3.54-3.47 (m, 1H), 3.47-3.40 (m, 1H), 3.29-3.23 (m, 1H), 3.17-3.10 (m, 1H), 2.57 (s, 3H), 1.96-1.84 (m, 4H), 1.73-1.68 (m, 2H), 1.51-1.47 (m, 10H), 1.23 (d,J= 6.3 Hz, 3H). ESI-MS m / z 641.0 [M+H + ]

[0236] Step 3) Preparation of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(1H-pyrrol-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (compound 7)

[0237] To a solution of 21 (7.1 mg, 0.011 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.03 mL, 0.110 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 7 (5.7 mg, 0.010 mmol, 91%) as a pale yellow solid.

[0238] 1 H NMR (500 MHz, MeOD) δ 8.53 (d,J= 7.2 Hz, 1H), 8.42 (s, 1H), 7.83 (d,J= 1.0 Hz, 1H), 7.28 (d,J= 3.5 Hz, 1H), 6.98 (d,J= 7.2 Hz, 1H), 6.74 (dd,J= 3.5, 1.8 Hz, 1H), 4.67 (s, 2H), 4.37-4.30 (m, 1H), 4.00-3.89 (m, 4H), 3.49 (d,J= 4.0 Hz, 1H), 3.25-3.17 (m, 1H), 3.17-3.10 (m, 1H), 2.57 (s, 3H), 2.03-1.97 (m, 2H), 1.96-1.90 (m, 1H), 1.80-1.75 (m, 1H), 1.35 (d,J= 6.5 Hz, 3H). ESI-MS m / z 540.9 [M+H + ]

[0239]

[0240] Compound 8. (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(thiophen-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride

[0241] Step 1) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-(thiophen-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (22)

[0242] To a solution of 8 (30 mg, 0.054 mmol, 1.0 eq) in ethanol (2.0 mL) at room temperature were added 2-(bromoacetyl)thiophene (13.3 mg, 0.065 mmol, 1.2 eq) and KI (1.8 mg, 0.011 mmol, 0.2 eq). The reaction mixture was stirred at 65 °C for 16 h. The reaction mass was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using DCM / MeOH (19 / 1) as an eluent to give 22 (10.5 mg, 0.016 mmol, 29%) as a white solid.

[0243] 1 H NMR (500 MHz, CDCl3) δ 8.01 (d,J= 6.5 Hz, 1H), 7.82 (s, 1H), 7.81-7.69 (m, 1H), 7.36 (d,J= 4.9 Hz, 1H), 7.15-7.10 (m, 1H), 6.61 (d,J= 6.9 Hz, 1H), 4.67-4.59 (m, 3H), 4.22-4.17 (m, 1H), 4.01 (dd,J= 10.6, 4.2 Hz, 1H), 3.75-3.69 (m, 2H), 3.55-3.48 (m, 1H), 3.46-3.39 (m, 1H), 3.30-3.22 (m, 1H), 3.17-3.10 (m, 1H), 2.56 (s, 3H), 1.96-1.82 (m, 4H), 1.73-1.67 (m, 1H), 1.48 (s, 9H), 1.23 (d,J= 6.3 Hz, 3H). ESI-MS m / z 657.0 [M+H + ]

[0244] Step 2) Preparation of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(thiophen-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (compound 8)

[0245] To a solution of 22 (10.5 mg, 0.016 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.04 mL, 0.160 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h. The solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for several minutes, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 8 (9.2 mg, 0.015 mmol, 97%) as a pale yellow solid.

[0246] 1 H NMR (500 MHz, MeOD) δ 8.52 (d,J= 7.2 Hz, 1H), 8.44 (s, 1H), 7.84 (d,J= 3.6 Hz, 1H), 7.77 (d,J= 5.0 Hz, 1H), 7.33-7.26 (m, 1H), 7.00 (d,J= 7.2 Hz, 1H), 4.67 (s, 2H), 4.37-4.28 (m, 1H), 4.05-3.96 (m, 2H), 3.95-3.87 (m, 2H), 3.50 (d,J= 4.0 Hz, 1H), 3.25-3.17 (m, 1H), 3.17-3.10 (m, 1H), 2.57 (s, 3H), 2.04-1.96 (m, 3H), 1.96-1.90 (m, 1H), 1.79-1.74 (m, 1H), 1.35 (d,J= 6.5 Hz, 3H). ESI-MS m / z 556.9 [M+H + ]

[0247]

[0248] Compound 9. (7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-2-methylimidazo[1,2-a]pyridin-3-yl)(piperazin-1-yl)methanone dihydrochloride

[0249] Step 1) Preparation of ethyl 2-bromo-3-oxobutanoate (23)

[0250] To a solution of ethyl acetoacetate (10.00 g, 76.84 mmol, 1.0 eq) in DCM (20 mL) at room temperature were added N-bromosuccinimide (13.68 g, 7.84 mmol, 1.0 eq) and pTsOHㆍH2O (2.92 g, 15.37 mmol, 0.2 eq). The reaction mixture was stirred at room temperature for 3 h. The reaction was washed with NaHCO 3(aq) , quenched with water and extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4 and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as the eluent to give 23 (6.87 g, 32.86 mmol, 43%) as a colorless oil.

[0251] 1 H NMR (400 MHz, CDCl3) δ 4.78 (s, 1H), 4.32 (q,J= 7.1 Hz, 2H), 2.47 (s, 3H), 1.34 (t,J= 7.1 Hz, 3H).

[0252] Step 2) Preparation of ethyl 8-chloro-7-iodo-2-methylimidazo[1,2-a]pyridine-3-carboxylate (24)

[0253] To a solution of 3-chloro-4-iodopyridin-2-amine (3.40 g, 13.36 mmol, 1.0 eq) in IPA (10 mL) and H2O (10 mL) at room temperature was added 23 (2.79 mg, 13.36 mmol, 1.0 eq). The reaction mixture was stirred at 95 °C for 2 h. The resulting solid was collected by filtration, washed with methanol, and dried under vacuum to give 24 (1.60 g, 4.39 mmol, 33%) as a red solid.

[0254] 1 H NMR (400 MHz, DMSO-d6) δ 8.90 (d,J= 7.2 Hz, 1H), 7.58 (d,J= 7.2 Hz, 1H), 4.38 (q,J= 7.1 Hz, 2H), 2.62 (s, 3H), 1.37 (t,J= 7.1 Hz, 3H). ESI-MS m / z 364.9 [M+H + ]

[0255] Step 3) Preparation of ethyl 8-chloro-7-((3-methoxy-3-oxopropyl)thio)-2-methylimidazo[1,2-a]pyridine-3-carboxylate (25)

[0256] To a solution of 24 (580 mg, 1.59 mmol, 1.0 eq) in 1,4-dioxane (10 mL) at room temperature were added methyl 3-mercaptopropionate (0.19 mL, 1.75 mmol, 1.1 eq), Pd2(dba)3 (73 mg, 0.08 mmol, 0.05 eq), Xantphos (92 mg, 0.16 mmol, 0.1 eq), and DIEA (0.86 mL, 4.30 mmol, 3.1 eq). The mixture was purged with N2 and evacuated three times. The reaction mixture was stirred at 90 °C for 2 h. The reaction was quenched with water and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (1 / 1) as eluent to give 25 (325 mg, 0.91 mmol, 57%) as a red solid.

[0257] 1 H NMR (400 MHz, DMSO) δ 8.59 (s, 1H), 8.54 (d,J= 7.3 Hz, 1H), 7.19 (d,J= 7.3 Hz, 1H), 3.86 (s, 3H), 3.62 (s, 3H), 3.39-3.35 (m, 2H), 2.73 (t,J= 7.0 Hz, 2H).

[0258] Step 4) Preparation of ethyl 7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-2-methylimidazo[1,2-a]pyridine-3-carboxylate (26)

[0259] To a solution of 25 (170 mg, 0.480 mmol, 1.0 eq) in 1,4-dioxane (3.0 mL) and DMF (1.0 mL) at room temperature were added 8 (273 mg, 0.580 mmol, 1.2 eq), Pd2(dba)3 (22 mg, 0.024 mmol, 0.05 eq), Xanthos (28 mg, 0.048 mmol, 0.1 eq), tBuOK (11 mg, 0.096 mmol, 2.0 eq), and DIEA (0.26 mL, 1.49 mmol, 3.1 eq). The mixture was purged with N2 and evacuated three times. The reaction mixture was stirred under microwave irradiation at 95 °C for 1 h. The reaction was quenched with water and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed under vacuum. The residue was purified by silica gel column chromatography using Hex / EtOAc (3 / 7) as the eluent to give 26 (171 mg, 0.260 mmol, 55%) as a yellow solid.

[0260] 1 H NMR (500 MHz, DMSO-d6) δ 8.97 (d,J= 7.4 Hz, 1H), 7.02 (d,J= 10.5 Hz, 1H), 6.60 (d,J= 7.4 Hz, 1H), 5.38 (t,J= 5.8 Hz, 1H), 4.45 (d,J= 5.7 Hz, 2H), 4.35 (q,J= 7.1 Hz, 2H), 4.22-4.13 (m, 1H), 3.90 (dd,J= 10.3, 5.0 Hz, 1H), 3.69 (d,J= 8.4 Hz, 1H), 3.53 (d,J= 8.4 Hz, 2H), 3.48-3.36 (m, 2H), 2.62 (s, 3H), 2.43 (s, 3H), 1.79-1.66 (m, 3H), 1.62-1.54 (m, 1H), 1.41 (s, 9H), 1.35 (t,J= 7.1 Hz, 3H), 1.31-1.27 (m, 1H), 1.03 (d,J= 6.3 Hz, 3H). ESI-MS m / z 661.2 [M+H+ ]

[0261] Step 5) Preparation of 7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-2-methylimidazo[1,2-a]pyridine-3-carboxylic acid (27)

[0262] To a solution of 26 (150 mg, 0.23 mmol, 1.0 eq) in THF (4.60 mL) and MeOH (1.15 mL) at room temperature was added 1N-LiOH (1.15 mL, 1.15 mmol, 5.0 eq) in H2O. The reaction mixture was stirred at room temperature for 1 h. TLC analysis showed the complete disappearance of the starting material. The reaction mixture was concentrated to remove THF and methanol. The aqueous layer containing the product was separated into 1N-HCl (aq) The pH was adjusted to 2 using and the mixture was extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo to give 27 (100 mg, 0.16 mmol, 68%) as a pale yellow solid.

[0263] 1H NMR (400 MHz, DMSO-d6) δ 9.02 (d,J= 7.4 Hz, 1H), 7.01 (d,J= 10.5 Hz, 1H), 6.60 (d,J= 7.4 Hz, 1H), 5.36 (t,J= 5.3 Hz, 1H), 4.50-4.40 (m, 2H), 4.22-4.12 (m, 1H), 3.89 (dd,J= 10.5, 5.0 Hz, 1H), 3.69 (d,J= 8.4 Hz, 1H), 3.59-3.49 (m, 2H), 3.46-3.37 (m, 2H), 3.30-3.24 (m, 1H), 2.61 (s, 3H), 2.44 (s, 3H), 1.80-1.65 (m, 3H), 1.63-1.53 ​​(m, 1H), 1.41 (s, 9H), 1.33-1.27 (m,J= 7.0 Hz, 1H), 1.03 (d,J= 6.3 Hz, 3H).

[0264] Step 6) Preparation of tert-butyl 4-(7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-2-methylimidazo[1,2-a]pyridine-3-carbonyl)piperazine-1-carboxylate (28)

[0265] To a solution of 27 (25 mg, 0.039 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 1-Boc-piperazine (11 mg, 0.059 mmol, 1.5 eq), HATU (22 mg, 0.059 mmol, 1.5 eq), and DIEA (15 μL, 0.078 mmol, 2.0 eq). After stirring at room temperature for 16 h, the reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 28 (28 mg, 0.035 mmol, 88%) as a yellow solid.

[0266] 1 H NMR (400 MHz, DMSO-d6) δ 8.25 (d,J= 7.3 Hz, 1H), 7.01 (d,J= 10.1 Hz, 1H), 6.47 (d,J= 7.3 Hz, 1H), 5.36 (t,J= 5.7 Hz, 1H), 4.44 (d,J= 5.4 Hz, 2H), 4.22-4.11 (m, 1H), 3.89 (dd,J= 10.5, 4.7 Hz, 1H), 3.68 (d,J= 8.5 Hz, 1H), 3.59-3.47 (m, 6H), 3.45-3.36 (m, 5H), 3.27-3.22 (m, 1H), 2.90 (t,J= 7.2 Hz, 1H), 2.43 (s, 3H), 2.39 (s, 3H), 1.78-1.64 (m, 3H), 1.61-1.53 ​​(m, 1H), 1.41 (d,J= 4.1 Hz, 18H), 1.03 (d,J= 6.3 Hz, 3H). ESI-MS m / z 801.7 [M+H + ]

[0267] Step 7) Preparation of (7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-2-methylimidazo[1,2-a]pyridin-3-yl)(piperazin-1-yl)methanone dihydrochloride (compound 9)

[0268] To a solution of 28 (25 mg, 0.031 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.08 mL, 0.31 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 9 (15 mg, 0.022 mmol, 64%) as a solid.

[0269] 1 H NMR (500 MHz, DMSO-d6) δ 9.46 (s, 2H), 8.33 (d, J= 7.4 Hz, 1H), 8.17 (s, 3H), 6.55 (s, 1H), 4.46 (s, 2H), 4.24-4.19 (m, 1H), 3.95-3.88 (m, 2H), 3.85-3.73 (m, 6H), 3.67 (d,J= 8.9 Hz, 1H), 3.22-3.11 (m, 5H), 3.10-3.00 (m, 2H), 2.44 (d,J= 12.4 Hz, 6H), 1.95-1.84 (m, 2H), 1.73 (dd,J= 12.9, 1.9 Hz, 1H), 1.63 (dd,J= 12.8, 1.5 Hz, 1H), 1.24 (d,J= 6.4 Hz, 3H). ESI-MS m / z 635.2 [M+H - ]

[0270]

[0271] Compound 10. (7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-2-methylimidazo[1,2-a]pyridin-3-yl)(4-aminopiperidin-1-yl)methanone dihydrochloride

[0272] Step 1) Preparation of tert-butyl ((3S,4S)-8-(5-((3-(4-((tert-butoxycarbonyl)amino)piperidine-1-carbonyl)-8-chloro-2-methylimidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (29)

[0273] To a solution of 27 (25 mg, 0.039 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 4-(tert-butoxycarbonylamino)piperidine (13 mg, 0.059 mmol, 1.5 eq), HATU (22 mg, 0.059 mmol, 1.5 eq), and DIEA (14 μL, 0.078 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 16 h, and the reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 29 (27 mg, 0.033 mmol, 84%) as a yellow solid.

[0274] 1 H NMR (400 MHz, DMSO-d6) δ 8.20 (d,J= 7.4 Hz, 1H), 7.01 (d,J= 10.6 Hz, 1H), 6.95-6.83 (m,J= 17.7, 6.9 Hz, 1H), 6.47 (d,J= 7.3 Hz, 1H), 4.44 (s, 2H), 4.19-4.13 (m, 1H), 3.89 (dd,J= 10.5, 4.9 Hz, 1H), 3.72-3.58 (m, 3H), 3.53 (d,J= 8.5 Hz, 2H), 3.44-3.38 (m, 2H), 3.28-3.23 (m, 1H), 3.19-3.06 (m, 4H), 2.92-2.84 (m, 3H), 2.43 (s, 3H), 2.37 (s, 3H), 1.81-1.69 (m, 4H), 1.60-1.54 (m, 1H), 1.42-1.37 (m, 18H), 1.27 (d,J= 5.7 Hz, 3H). ESI-MS m / z 815.4 [M+H + ]

[0275] Step 2) Preparation of (7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-2-methylimidazo[1,2-a]pyridin-3-yl)(4-aminopiperidin-1-yl)methanone dihydrochloride (Compound 10)

[0276] To a solution of 29 (24 mg, 0.029 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.07 mL, 0.29 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 10 (13 mg, 0.019 mmol, 52%) as a solid.

[0277] 1 H NMR (400 MHz, MeOD) δ 8.47 (d,J= 4.6 Hz, 1H), 6.93 (d,J= 7.4 Hz, 1H), 4.65 (s, 2H), 4.34 -4.31 (m, 1H), 4.01-3.98 (m, 1H), 3.97-3.91 (m, 1H), 3.91-3.84 (m, 2H), 3.51-3.46 (m, 2H), 3.23-3.09 (m, 3H), 3.08 (s, 1H), 3.00 (d,J= 2.7 Hz, 2H), 2.97 (s, 1H), 2.59 (s, 3H), 2.55 (d,J= 5.8 Hz, 3H), 2.17-2.11 (m, 2H), 2.02-1.92 (m, 3H), 1.79-1.71 (m, 2H), 1.34 (d,J= 6.5 Hz, 3H). ESI-MS m / z 649.2 [M+H - ]

[0278]

[0279] Compound 11. 7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-N-(2-methoxyethyl)-2-methylimidazo[1,2-a]pyridine-3-carboxamide hydrochloride

[0280] Step 1) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-3-((2-methoxyethyl)carbamoyl)-2-methylimidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (30)

[0281] To a solution of 27 (25 mg, 0.039 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 2-methoxyethylamine (5.1 μL, 0.059 mmol, 1.5 eq), HATU (22 mg, 0.059 mmol, 1.5 eq), and DIEA (14 μL, 0.078 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 16 h, and the reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 30 (13 mg, 0.019 mmol, 48%) as a yellow solid.

[0282] ESI-MS m / z 690.3 [M+H + ]

[0283] Step 2) Preparation of 7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-N-(2-methoxyethyl)-2-methylimidazo[1,2-a]pyridine-3-carboxamide hydrochloride (Compound 11)

[0284] To a solution of 30 (10 mg, 0.014 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.04 mL, 0.140 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 11 (2.3 mg, 0.004 mmol, 26%) as a pale yellow solid.

[0285] 1 H NMR (400 MHz, DMSO-d6) δ 8.77 (d,J= 7.4 Hz, 1H), 8.10-8.02 (m, 3H), 6.59 (d,J= 7.4 Hz, 1H), 4.46 (s, 2H), 4.24-4.18 (m, 1H), 3.93-3.80 (m, 4H), 3.70-3.67 (m, 2H), 3.49-3.46 (m, 5H), 3.29 (s, 3H), 3.12-3.00 (m, 2H), 2.58 (s, 3H), 2.44 (s, 3H), 1.92-1.83 (m, 2H), 1.77-1.71 (m, 1H), 1.66-1.59 (m, 1H), 1.23 (d,J= 6.3 Hz, 3H).

[0286]

[0287] Compound 12. 7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-2-methyl-N-(piperidin-4-yl)imidazo[1,2-a]pyridine-3-carboxamide dihydrochloride

[0288] Step 1) Preparation of tert-butyl 4-(7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-2-methylimidazo[1,2-a]pyridine-3-carboxamido)piperidine-1-carboxylate (31)

[0289] To a solution of 27 (25 mg, 0.039 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 4-amino-1-Boc-piperidine (12 mg, 0.059 mmol, 1.5 eq), HATU (22 mg, 0.059 mmol, 1.5 eq), and DIEA (14 μL, 0.078 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 16 h, and the reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 31 (15 mg, 0.018 mmol, 58%) as a yellow solid.

[0290] 1H NMR (400 MHz, DMSO-d6) δ 8.68 (d,J= 7.5 Hz, 1H), 7.89 (d,J= 8.0 Hz, 1H), 7.01 (d,J= 10.2 Hz, 1H), 6.50 (d,J= 7.3 Hz, 1H), 5.34 (t,J= 5.7 Hz, 1H), 4.44 (d,J= 5.4 Hz, 2H), 4.21-4.13 (m, 1H), 3.96-3.86 (m, 3H), 3.69 (d,J= 8.8 Hz, 1H), 3.65-3.58 (m, 1H), 3.53 (d,J=8.3 Hz, 2H), 3.45-3.37 (m, 2H), 3.29-3.20 (m, 2H), 3.18-3.12 (m, 1H), 2.55 (s, 3H), 2.43 (s, 3H), 1.86-1.80 (m, 2H), 1.75-1.65 (m, 3H), 1.60-1.53 ​​(m, 1H), 1.43-1.38 (m, 20H), 1.03 (d,J= 6.3 Hz, 3H). ESI-MS m / z 815.4 [M+H + ]

[0291] Step 2) Preparation of 7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-2-methyl-N-(piperidin-4-yl)imidazo[1,2-a]pyridine-3-carboxamide dihydrochloride (Compound 12)

[0292] To a solution of 31 (12 mg, 0.015 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.04 mL, 0.150 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h. The solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 12 (6.5 mg, 0.009 mmol, 65%) as a pale yellow solid.

[0293] 1 H NMR (400 MHz, DMSO-d6) δ 8.97-8.86 (m, 1H), 8.86-8.71 (m, 1H), 8.67 (d,J= 7.3 Hz, 1H), 8.26 (d,J= 7.0 Hz, 1H), 8.21-8.05 (m, 3H), 6.58 (d,J= 7.4 Hz, 1H), 4.46 (s, 2H), 4.26-4.18 (m, 1H), 4.12-4.05 (m, 1H), 3.92-3.87 (m, 2H), 3.84-3.77 (m, 2H), 3.42-3.39 (m, 1H), 3.33-3.28 (m, 2H), 3.09-2.99 (m, 4H), 2.57 (s, 3H), 2.44 (s, 3H), 2.04-1.98 (m, 2H), 1.94-1.71 (m, 6H), 1.65-1.58 (m, 1H), 1.24 (d,J= 6.1 Hz, 3H).

[0294]

[0295] Compound 13. (7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)(piperazin-1-yl)methanone dihydrochloride

[0296] Step 1) Preparation of ethyl 7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridine-2-carboxylate (32)

[0297] To a solution of 8 (350 mg, 0.64 mmol, 1.0 eq) in ethanol (5.0 mL) at room temperature was added ethyl 3-bromo-2-oxopropanoate (0.09 mL, 0.70 mmol, 1.1 eq). After stirring at 75 °C for 3 h, the reaction was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using DCM / MeOH (19 / 1) as the eluent to give 32 (150 mg, 0.23 mmol, 36%) as a pale yellow solid.

[0298] 1 H NMR (400 MHz, DMSO-d6) δ 8.60 (s, 1H), 8.38 (d,J= 7.3 Hz, 1H), 7.01 (d,J= 10.5 Hz, 1H), 6.51 (d,J= 7.3 Hz, 1H), 5.35 (t,J= 5.7 Hz, 1H), 4.44 (d,J= 5.7 Hz, 2H), 4.33 (q,J= 7.1 Hz, 2H), 4.22-4.12 (m, 1H), 3.89 (dd,J= 10.5, 4.9 Hz, 1H), 3.68 (d,J= 8.3 Hz, 1H), 3.57-3.47 (m,J=8.4 Hz, 2H), 3.45-3.37 (m, 2H), 3.29-3.20 (m,J= 8.1 Hz, 1H), 2.44 (s, 3H), 1.79-1.64 (m, 3H), 1.62-1.51 (m,J= 13.6 Hz, 1H), 1.40 (s, 9H), 1.33 (t,J= 7.1 Hz, 3H), 1.03 (d,J= 6.3 Hz, 3H). ESI-MS m / z 647.2 [M+H + ]

[0299] Step 2) Preparation of 7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridine-2-carboxylic acid (33)

[0300] To a solution of 32 (150 mg, 0.32 mmol, 1.0 eq) in THF (6.40 mL) and MeOH (1.60 mL) at room temperature was added 1N-LiOH (1.60 mL, 1.60 mmol, 5.0 eq) in H2O. The reaction mixture was stirred at room temperature for 1 h. After TLC analysis showed the complete disappearance of the starting material, the reaction mixture was concentrated to remove THF and methanol. The residue was purified by filtration with 1N-HCl (aq) The pH was adjusted to 2 using and extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo to give 33 (120 mg, 0.19 mmol, 84%) as a pale yellow solid.

[0301] 1 H NMR (400 MHz, DMSO-d6) δ 8.52 (s, 1H), 8.38 (d,J= 7.3 Hz, 1H), 7.01 (d,J= 10.5 Hz, 1H), 6.50 (d,J= 7.3 Hz, 1H), 5.35 (d,J= 5.2 Hz, 1H), 4.44 (d,J= 4.5 Hz, 2H), 4.19-4.13 (m, 1H), 3.89 (dd,J= 10.4, 5.0 Hz, 1H), 3.68 (d,J= 8.5 Hz, 1H), 3.57-3.42 (m, 4H), 3.28-3.21 (m, 2H), 2.44 (s, 3H), 1.78-1.65 (m, 3H), 1.61-1.52 (m, 1H), 1.40 (s, 9H), 1.03 (d,J= 6.3 Hz, 3H).ESI-MS m / z 619.2 [M+H + ]

[0302] Step 3) Preparation of tert-butyl 4-(7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridine-2-carbonyl)piperazine-1-carboxylate (34)

[0303] To a solution of 33 (20 mg, 0.032 mmol, 1.0 eq) in DMF (0.5 mL) at room temperature were added 1-Boc-piperazine (8.9 mg, 0.048 mmol, 1.5 eq), HATU (18 mg, 0.048 mmol, 1.5 eq), and DIEA (11 μL, 0.064 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 34 (22 mg, 0.028 mmol, 88%) as a yellow solid.

[0304] 1 H NMR (500 MHz, DMSO-d6) δ 8.39 (d,J= 8.2 Hz, 2H), 7.01 (d,J= 10.5 Hz, 1H), 6.50 (d,J= 7.2 Hz, 1H), 5.35 (t,J= 5.6 Hz, 1H), 4.44 (d,J= 5.3 Hz, 2H), 4.21-4.06 (m,3H), 3.89 (dd,J= 10.4, 5.0 Hz, 1H), 3.68 (d,J= 8.3 Hz, 1H), 3.67-3.57 (m, 2H), 3.53 (d,J= 8.4 Hz, 1H), 3.52-3.45 (m, 1H), 3.45-3.35 (m, 6H), 3.31-3.20 (m, 2H), 2.43 (s, 3H), 1.78-1.63 (m, 3H), 1.62-1.54 (m, 1H), 1.42 (d,J= 11.5 Hz, 17H), 1.03 (d,J= 6.2 Hz, 3H). ESI-MS m / z 787.2 [M+H + ]

[0305] Step 4) Preparation of (7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)(piperazin-1-yl)methanone dihydrochloride (compound 13)

[0306] To a solution of 34 (20 mg, 0.025 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.06 mL, 0.250 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 16 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 13 (11 mg, 0.017 mmol, 65%) as a pale yellow solid.

[0307] 1 H NMR (500 MHz, DMSO-d6) δ 9.30-9.15 (m, 2H), 8.46 (s, 1H), 8.43-8.42 (m, 1H), 8.23-8.01 (m, 3H), 6.57-6.48 (m, 1H), 4.57-4.33 (m, 4H), 4.25-4.17 (m, 1H), 3.99-3.77 (m, 5H), 3.70-3.65 (m, 1H), 3.24-3.17 (m, 4H), 3.10-2.98 (m, 2H), 2.50-2.40 (m, 4H), 1.96-1.84 (m, 2H), 1.79-1.70 (m, 1H), 1.68-1.59 (m, 1H), 1.29-1.19 (m, 3H). ESI-MS m / z 587.1 [M+H + ]

[0308]

[0309] Compound 14. (7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)(4-aminopiperidin-1-yl)methanone dihydrochloride

[0310] Step 1) Preparation of tert-butyl ((3S,4S)-8-(5-((2-(4-((tert-butoxycarbonyl)amino)piperidine-1-carbonyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (35)

[0311] To a solution of 33 (25 mg, 0.04 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 4-(tert-butoxycarbonylamino)piperidine (13 mg, 0.06 mmol, 1.5 eq), HATU (23 mg, 0.06 mmol, 1.5 eq), and DIEA (14 μL, 0.08 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 35 (24 mg, 0.030 mmol, 75%) as a yellow solid.

[0312] 1H NMR (400 MHz, DMSO-d6) δ 8.38 (d,J= 7.3 Hz, 1H), 8.32 (s, 1H), 7.01 (d,J= 10.5 Hz, 1H), 6.90 (d,J= 7.9 Hz, 1H), 6.48 (d,J= 7.2 Hz, 1H), 5.35 (t,J= 5.8 Hz, 1H), 4.91-4.77 (m, 1H), 4.44 (d,J= 5.7 Hz, 2H), 4.41-4.27 (m, 1H), 4.22-4.12 (m, 1H), 3.89 (dd,J= 10.5, 4.8 Hz, 1H), 3.68 (d,J= 8.4 Hz, 1H), 3.59-3.47 (m, 3H), 3.44-3.38 (m, 3H), 3.31-3.21 (m, 4H), 2.43 (s, 3H), 1.87-1.53 ​​(m, 7H), 1.40 (d,J= 3.9 Hz, 18H), 1.03 (d,J= 6.3 Hz, 3H). ESI-MS m / z 801.6 [M+H + ]

[0313] Step 2) Preparation of (7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)(4-aminopiperidin-1-yl)methanone dihydrochloride (Compound 14)

[0314] To a solution of 35 (20 mg, 0.025 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.13 mL, 0.500 mmol, 20 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 16 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 14 (13 mg, 0.019 mmol, 70%) as a pale yellow solid.

[0315] 1H NMR (400 MHz, DMSO-d6) δ 8.42 (d,J= 7.2 Hz, 1H), 8.38 (s, 1H), 8.18-8.06 (m, 6H), 6.52 (d,J= 7.2 Hz, 1H), 5.07-4.93 (m, 1H), 4.57-4.48 (m, 1H), 4.46 (s, 2H), 4.25-4.18 (m, 1H), 3.87 (dd,J= 11.6, 6.2 Hz, 2H), 3.84-3.77 (m, 1H), 3.68 (d,J= 9.0 Hz, 1H), 3.24-3.20 (m, 1H), 3.12-2.89 (m, 4H), 2.45 (s, 3H), 2.06-2.00 (m, 2H), 1.96-1.82 (m, 3H), 1.78-1.72 (m, 1H), 1.66-1.42 (m, 5H). ESI-MS m / z 601.1 [M+H + ]

[0316]

[0317] Compound 15. 7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-N-(piperidin-4-yl)imidazo[1,2-a]pyridine-2-carboxamide dihydrochloride

[0318] Step 1) Preparation of tert-butyl 4-(7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridine-2-carboxamido)piperidine-1-carboxylate (36)

[0319] To a solution of 33 (25 mg, 0.04 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 4-amino-1-Boc-piperidine (12 mg, 0.06 mmol, 1.5 eq), HATU (23 mg, 0.06 mmol, 1.5 eq), and DIEA (14 μL, 0.08 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 36 (22 mg, 0.027 mmol, 69%) as a yellow solid.

[0320] 1 H NMR (400 MHz, DMSO-d6) δ 8.41 (s, 1H), 8.39 (d,J= 7.2 Hz, 1H), 8.13 (d,J= 8.6 Hz, 1H), 7.01 (d,J= 10.5 Hz, 1H), 6.49 (d,J= 7.2 Hz, 1H), 5.34 (t,J= 5.7 Hz, 1H), 4.44 (d,J= 5.6 Hz, 2H), 4.22-4.13 (m, 1H), 4.02-3.92 (m, 3H), 3.92-3.86 (m, 1H), 3.68 (d,J= 8.4 Hz, 1H), 3.56-3.48 (m, 2H), 3.43-3.38 (m, 2H), 3.29-3.20 (m, 1H), 2.44 (s, 3H), 1.80-1.67 (m, 5H), 1.63-1.51 (m, 3H), 1.46-1.36 (m, 20H), 1.03 (d,J=6.3 Hz, 3H). ESI-MS m / z 801.3 [M+H + ]

[0321] Step 2) Preparation of 7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-N-(piperidin-4-yl)imidazo[1,2-a]pyridine-2-carboxamide dihydrochloride (Compound 15)

[0322] To a solution of 36 (18 mg, 0.022 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.11 mL, 0.44 mmol, 20 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 16 h. The solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 15 (5.2 mg, 0.008 mmol, 35%) as a pale yellow solid.

[0323] 1 H NMR (500 MHz, MeOD) δ 8.47 (s, 1H), 8.41 (d,J= 7.2 Hz, 1H), 6.73 (d,J= 7.2 Hz, 1H), 4.88-4.86 (m, 2H), 4.64 (s, 2H), 4.33 (dd,J= 6.5, 4.1 Hz, 1H), 4.28-4.21 (m, 1H), 3.99 (d,J= 9.2 Hz, 1H), 3.95-3.81 (m, 3H), 3.55-3.47 (m, 3H), 3.26-3.15 (m, 3H), 3.14-3.03 (m, 2H), 2.26 (dd,J= 14.3, 3.3 Hz, 2H), 2.03-1.90 (m, 5H), 1.75 (d,J= 13.0 Hz, 1H), 1.35 (d,J= 6.5 Hz, 3H). ESI-MS m / z 601.1 [M+H + ]

[0324]

[0325] Compound 16. 7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-N-(2-methoxyethyl)imidazo[1,2-a]pyridine-2-carboxamide hydrochloride

[0326] Step 1) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-((2-methoxyethyl)carbamoyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (37)

[0327] To a solution of 33 (25 mg, 0.04 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 2-methoxyethylamine (5.2 μL, 0.06 mmol, 1.5 eq), HATU (23 mg, 0.06 mmol, 1.5 eq), and DIEA (14 μL, 0.08 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 37 (12 mg, 0.033 mmol, 44%) as a yellow solid.

[0328] 1H NMR (400 MHz, DMSO-d6) δ 8.46-8.35 (m, 2H), 8.21-8.10 (m, 1H), 7.01 (d,J= 10.6 Hz, 1H), 6.49 (d,J= 7.2 Hz, 1H), 5.39-5.29 (m, 1H), 4.44 (s, 2H), 4.21-4.11 (m, 1H), 3.89 (dd,J= 10.6, 5.0 Hz, 1H), 3.68 (d,J= 8.4 Hz, 1H), 3.55-3.38 (m, 8H), 3.28 (s, 3H), 2.44 (s, 3H), 1.80-1.66 (m, 3H), 1.62-1.53 ​​(m, 1H), 1.40 (s, 9H), 1.03 (d,J= 6.3 Hz, 3H). ESI-MS m / z 676.1 [M+H + ]

[0329] Step 2) Preparation of 7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-N-(2-methoxyethyl)imidazo[1,2-a]pyridine-2-carboxamide hydrochloride (Compound 16)

[0330] To a solution of 37 (10 mg, 0.015 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.08 mL, 0.30 mmol, 20 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 16 (2.5 mg, 0.004 mmol, 27%) as a pale yellow solid.

[0331] 1H NMR (500 MHz, MeOD) δ 8.31 (s, 1H), 8.30 (d,J= 7.3 Hz, 1H), 6.59 (d,J= 7.2 Hz, 1H), 4.62 (s, 2H), 4.32-4.26 (m, 1H), 3.93 (d,J= 8.9 Hz, 1H), 3.81 (d,J= 9.0 Hz, 1H), 3.80-3.71 (m, 2H), 3.64-3.60 (m, 4H), 3.44-3.41 (m, 3H), 3.27 (d,J= 4.4 Hz, 1H), 3.22-3.16 (m, 1H), 3.14-3.08 (m, 1H), 2.54 (s, 3H), 1.99-1.90 (m, 2H), 1.86-1.81 (m, 1H), 1.75-1.70 (m, 1H), 1.29 (d,J= 6.5 Hz, 3H). ESI-MS m / z 576.1 [M+H + ]

[0332]

[0333] Compound 17. 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(piperazin-1-yl)ethanone dihydrochloride

[0334] Step 1) Preparation of methyl 2-(4-acetylphenoxy)acetate (38)

[0335] To a solution of 4-hydroxyacetonephenone (1.30 g, 9.55 mmol, 1.0 eq) in acetone (20 mL) at room temperature were added methyl bromoacetate (1.08 mL, 11.46 mmol, 1.2 eq) and K2CO3 (1.58 g, 11.46 mmol, 1.2 eq). The reaction mixture was stirred at 80 °C for 3 h. The reaction was quenched with water and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (7 / 3) as an eluent to give 38 (1.80 g, 8.65 mmol, 91%) as a white solid.

[0336] 1 H NMR (500 MHz, CDCl3) δ 8.00-7.91 (m, 2H), 7.03-6.89 (m, 2H), 4.73 (s, 2H), 3.84 (s, 3H), 2.58 (s, 3H).

[0337] Step 2) Preparation of methyl 2-(4-(2-bromoacetyl)phenoxy)acetate (39)

[0338] To a solution of 38 (1.20 g, 5.76 mmol, 1.0 eq) in chloroform (30 mL) at room temperature were added N-bromosuccinimide (1.23 g, 6.91 mmol, 1.2 eq) and pTSAㆍH2O (219 mg, 1.15 mmol, 0.2 eq). The reaction mixture was stirred at 70 °C for 5 h. The reaction was quenched with NaHCO3 solution and water, then extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (7 / 3) as an eluent to give 39 (1.32 g, 4.60 mmol, 80%) as a white solid.

[0339] 1H NMR (500 MHz, CDCl3) δ 8.00 (d,J= 8.9 Hz, 2H), 6.99 (d,J= 8.9 Hz, 2H), 4.74 (s, 2H), 4.42 (s, 2H), 3.85 (s, 3H). ESI-MS m / z 286.9 [M+H + ]

[0340] Step 3) Preparation of methyl 2-(4-(7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetate (40)

[0341] To a solution of 8 (500 mg, 0.91 mmol, 1.0 eq) in ethanol (20 mL) at room temperature were added 39 (314 mg, 1.09 mmol, 1.2 eq) and KI (30 mg, 0.18 mmol, 0.2 eq). The reaction mixture was stirred at 65 °C for 5 h, and the reaction mass was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using DCM / MeOH (19 / 1) as an eluent to give 40 (210 mg, 0.24 mmol, 31%) as a pale yellow solid.

[0342] ESI-MS m / z 739.6 [M+H + ]

[0343] Step 4) Preparation of 2-(4-(7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetic acid (41)

[0344] To a solution of 40 (210 mg, 0.28 mmol, 1.0 eq) in THF (4.56 mL) and MeOH (1.14 mL) at room temperature was added 1N-LiOH (1.14 mL, 1.14 mmol, 4.0 eq) dissolved in H2O. The reaction mixture was stirred at room temperature for 2 h. After TLC analysis showed the complete disappearance of the starting material, the reaction mixture was concentrated to remove THF and methanol. The reaction mass was purified by filtration using 1N-HCl (aq) The pH was adjusted to 2 using and extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4 and the solvent was removed under vacuum to give 41 (150 mg, 0.17 mmol, 73%) as a pale yellow solid.

[0345] 1 H NMR (500 MHz, DMSO-d6) δ 8.40-8.31 (m, 2H), 7.89 (d,J= 8.8 Hz, 2H), 7.04-6.97 (m, 3H), 6.40 (d,J= 7.1 Hz, 1H), 5.35 (t,J= 5.6 Hz, 1H), 4.73 (s, 2H), 4.44 (d,J= 5.3 Hz, 2H), 4.21-4.15 (m, 1H), 3.89 (dd,J= 10.4, 5.0 Hz, 1H), 3.69 (d,J= 8.4 Hz, 1H), 3.56-3.49 (m, 2H), 3.43-3.40 (m, 1H), 3.28-3.20 (m, 2H), 2.44 (s, 3H), 1.78-1.66 (m, 3H), 1.62-1.55 (m, 1H), 1.49-1.35 (m, 10H), 1.03 (d,J= 6.3 Hz, 3H). ESI-MS m / z 725.0 [M+H + ]

[0346] Step 5) Preparation of tert-butyl 4-(2-(4-(7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetyl)piperazine-1-carboxylate (42)

[0347] To a solution of 41 (25 mg, 0.034 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 1-Boc-piperazine (9.5 mg, 0.051 mmol, 1.5 eq), HATU (19 mg, 0.051 mmol, 1.5 eq), and DIEA (12 μL, 0.068 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 42 (26 mg, 0.029 mmol, 86%) as a yellow solid.

[0348] 1H NMR (400 MHz, DMSO-d6) δ 8.35 (d,J= 7.3 Hz, 2H), 7.88 (d,J= 8.9 Hz, 2H), 7.06-6.97 (m, 3H), 6.40 (d,J= 7.2 Hz, 1H), 4.89 (s, 2H), 4.44 (s, 2H), 4.20-4.13 (m, 1H), 3.89 (dd,J= 10.3, 4.8 Hz, 1H), 3.69 (d,J= 8.4 Hz, 1H), 3.53 (d,J= 8.5 Hz, 1H), 3.50-3.44 (m, 5H), 3.39 (s, 3H), 3.25-3.22 (m, 1H), 2.92 (s, 2H), 2.88 (s, 2H), 2.44 (s, 3H), 1.77-1.65 (m, 3H), 1.60-1.54 (m, 1H), 1.43-1.40 (m,J= 4.3 Hz, 18H), 1.03 (d,J= 6.2 Hz, 3H). ESI-MS m / z 893.1 [M+H - ]

[0349] Step 6) Preparation of 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(piperazin-1-yl)ethanone dihydrochloride (Compound 17)

[0350] To a solution of 42 (22 mg, 0.025 mmol, 1.0 eq) in DCM (10 mL) at 0 °C was added 4N-HCl (0.13 mL, 0.500 mmol, 20 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 17 (7.1 mg, 0.009 mmol, 38%) as a pale yellow solid.

[0351] 1H NMR (500 MHz, MeOD) δ 8.26 (d,J= 7.1 Hz, 1H), 8.16 (s, 1H), 7.92 (d,J= 8.8 Hz, 2H), 7.10 (d,J= 8.9 Hz, 2H), 6.59 (d,J= 7.1 Hz, 1H), 4.92 (s, 2H), 4.60 (s, 2H), 4.35-4.27 (m, 2H), 3.97 (d,J= 9.1 Hz, 1H), 3.86 (d,J= 9.1 Hz, 1H), 3.82-3.75 (m, 5H), 3.42 (d,J= 4.2 Hz, 1H), 3.20-3.15 (m, 2H), 3.14-3.10 (m, 2H), 3.08-3.00 (m, 1H), 2.55 (s, 3H), 2.05-1.94 (m, 3H), 1.91-1.86 (m, 1H), 1.75-1.72 (m, 1H), 1.33 (d,J=6.5 Hz, 3H). ESI-MS m / z 726.8 [M+H - ]

[0352]

[0353] Compound 18. 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(4-aminopiperidin-1-yl)ethanone dihydrochloride

[0354] Step 1) Preparation of tert-butyl ((3S,4S)-8-(5-((2-(4-(2-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-2-oxoethoxy)phenyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (43)

[0355] To a solution of 41 (25 mg, 0.034 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 4-(tert-butoxycarbonylamino)piperidine (11 mg, 0.051 mmol, 1.5 eq), HATU (19 mg, 0.051 mmol, 1.5 eq), and DIEA (12 μL, 0.068 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as eluent to give 43 (27 mg, 0.030 mmol, 88%) as a yellow solid.

[0356] 1 H NMR (500 MHz, DMSO-d6) δ 8.35 (d,J= 7.1 Hz, 2H), 7.88 (d,J= 8.7 Hz, 2H), 7.06-6.98 (m, 3H), 6.92 (d,J= 7.6 Hz, 1H), 6.40 (d,J= 7.1 Hz, 1H), 4.86 (q,J= 14.3 Hz, 2H), 4.44 (s, 2H), 4.23-4.14 (m, 2H), 3.89 (dd,J= 10.4, 4.9 Hz, 1H), 3.84-3.78 (m, 1H), 3.69 (d,J= 8.4 Hz, 1H), 3.56-3.49 (m, 3H), 3.44-3.38 (m, 2H), 3.28-3.23 (m, 1H), 3.18-3.09 (m, 2H), 2.91 (s, 1H), 2.87 (d,J= 4.8 Hz, 2H), 2.44 (s, 3H), 1.82-1.68 (m, 5H), 1.61-1.56 (m, 1H), 1.40 (d,J= 5.4 Hz, 18H), 1.03 (d,J= 6.2 Hz, 3H). ESI-MS m / z 907.1 [M+H + ]

[0357] Step 2) Preparation of 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(4-aminopiperidin-1-yl)ethanone dihydrochloride (Compound 18)

[0358] To a solution of 43 (22 mg, 0.024 mmol, 1.0 eq) in DCM (10 mL) at 0 °C was added 4N-HCl (0.12 mL, 0.48 mmol, 20 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 18 (5.3 mg, 0.007 mmol, 28%) as a pale yellow solid.

[0359] 1H NMR (500 MHz, MeOD) δ 8.25 (d,J= 7.1 Hz, 1H), 8.16 (s, 1H), 7.95-7.88 (m, 2H), 7.09 (d,J= 8.9 Hz, 2H), 6.56 (d,J= 7.1 Hz, 1H), 4.98-4.91 (m, 2H), 4.60 (s, 3H), 4.30-4.26 (m, 1H), 4.18-4.11 (m, 1H), 3.92 (d,J= 8.9 Hz, 1H), 3.80 (d,J= 8.9 Hz, 1H), 3.78-3.69 (m, 2H), 3.42-3.34 (m, 2H), 3.29-3.22 (m, 2H), 3.21-3.15 (m, 1H), 3.13-3.07 (m, 1H), 2.86-2.79 (m, 1H), 2.14-2.06 (m, 2H), 2.00-1.89 (m, 2H), 1.85-1.79 (m, 1H), 1.75-1.71 (m, 1H), 1.68-1.58 (m, 2H), 1.54-1.43 (m, 2H), 1.29 (d,J= 6.5 Hz, 3H). ESI-MS m / z 740.9 [M+H - ]

[0360]

[0361] Compound 19. 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(piperidin-4-yl)acetamide dihydrochloride

[0362] Step 1) Preparation of tert-butyl 4-(2-(4-(7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetamido)piperidine-1-carboxylate (44)

[0363] To a solution of 41 (25 mg, 0.034 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 4-amino-1-Boc-piperidine (10 mg, 0.051 mmol, 1.5 eq), HATU (19 mg, 0.051 mmol, 1.5 eq), and DIEA (12 μL, 0.068 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 44 (25 mg, 0.028 mmol, 81%) as a yellow solid.

[0364] 1 H NMR (400 MHz, DMSO-d6) δ 8.41-8.30 (m, 2H), 8.05 (d,J= 8.1 Hz, 1H), 7.91 (d,J= 8.8 Hz, 2H), 7.10-6.96 (m, 3H), 6.41 (d,J= 7.2 Hz, 1H), 4.52 (s, 2H), 4.44 (s, 2H), 4.20-4.13 (m, 1H), 3.94-3.83 (m, 4H), 3.69 (d,J= 8.5 Hz, 1H), 3.66-3.59 (m, 1H), 3.56-3.49 (m, 2H), 3.25-3.12 (m, 2H), 2.92-2.89 (m, 5H), 2.44 (s, 3H), 1.85-1.79 (m, 1H), 1.75-1.66 (m, 4H), 1.61-1.53 ​​(m, 1H), 1.40 (d,J= 2.1 Hz, 18H), 1.03 (d,J= 6.2 Hz, 3H). ESI-MS m / z 907.1 [M+H + ]

[0365] Step 2) Preparation of 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(piperidin-4-yl)acetamide dihydrochloride (Compound 19)

[0366] To a solution of 44 (20 mg, 0.022 mmol, 1.0 eq) in DCM (10 mL) at 0 °C was added 4N-HCl (0.11 mL, 0.440 mmol, 20 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 19 (3.4 mg, 0.004 mmol, 20%) as a pale yellow solid.

[0367] 1 H NMR (500 MHz, MeOD) δ 8.52 (d,J= 7.2 Hz, 1H), 8.40 (s, 1H), 7.93 (d,J= 8.8 Hz, 2H), 7.23 (d,J= 8.9 Hz, 2H), 6.94 (d,J= 7.2 Hz, 1H), 4.67 (s, 3H), 4.37-4.30 (m, 1H), 4.18-4.05 (m, 2H), 4.04-3.94 (m, 2H), 3.93-3.82 (m, 2H), 3.51-3.44 (m, 3H), 3.37 (s, 1H), 3.22-3.09 (m, 4H), 3.05 (s, 2H), 2.18-2.11 (m, 2H), 2.03-1.96 (m, 2H), 1.95-1.84 (m, 3H), 1.80-1.73 (m, 1H), 1.35 (d,J= 6.5 Hz, 3H). ESI-MS m / z 740.8 [M+H - ]

[0368]

[0369] Compound 20. 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(2-methoxyethyl)acetamide hydrochloride

[0370] Step 1) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-(4-(2-((2-methoxyethyl)amino)-2-oxoethoxy)phenyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (45)

[0371] To a solution of 41 (25 mg, 0.034 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 2-methoxyethylamine (4.4 μL, 0.051 mmol, 1.5 eq), HATU (19 mg, 0.051 mmol, 1.5 eq), and DIEA (12 μL, 0.068 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 45 (20 mg, 0.026 mmol, 74%) as a yellow solid.

[0372] 1H NMR (400 MHz, DMSO-d6) δ 8.40-8.30 (m, 2H), 8.14 (t,J= 5.6 Hz, 1H), 7.91 (d,J= 8.8 Hz, 2H), 7.05 (d,J= 8.9 Hz, 2H), 7.01 (d,J= 10.5 Hz, 1H), 6.40 (d,J= 7.1 Hz, 1H), 5.34 (s, 1H), 4.54 (s, 2H), 4.44 (s, 2H), 4.20-4.14 (m, 1H), 3.89 (dd,J= 10.5, 5.0 Hz, 1H), 3.69 (d,J=8.4 Hz, 1H), 3.56-3.49 (m, 2H), 3.41-3.38 (m, 4H), 3.31-3.27 (m, 2H), 3.25 (s, 3H), 2.93-2.87 (m, 1H), 2.44 (s, 3H), 1.78-1.64 (m, 3H), 1.61-1.53 ​​(m, 1H), 1.41 (s, 9H), 1.03 (d,J= 6.3 Hz, 3H). ESI-MS m / z 782.0 [M+H + ]

[0373] Step 2) Preparation of 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(2-methoxyethyl)acetamide hydrochloride (Compound 20)

[0374] To a solution of 45 (16 mg, 0.020 mmol, 1.0 eq) in DCM (10 mL) at 0 °C was added 4N-HCl (0.10 mL, 0.410 mmol, 20 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 20 (4.1 mg, 0.006 mmol, 28%) as a yellow solid.

[0375] 1 H NMR (500 MHz, MeOD) δ 8.27 (d,J= 7.1 Hz, 1H), 8.17 (s, 1H), 7.93 (d,J= 8.8 Hz, 2H), 7.11 (d,J= 8.8 Hz, 2H), 6.60 (d,J= 7.1 Hz, 1H), 4.61 (s, 2H), 4.60 (s, 2H), 4.34-4.29 (m, 1H), 3.98 (d,J= 9.1 Hz, 1H), 3.87 (d,J= 9.3 Hz, 1H), 3.86-3.81 (m, 1H), 3.79-3.74 (m, 1H), 3.52-3.49 (m, 4H), 3.48-3.45 (m, 1H), 3.36 (s, 3H), 3.16-3.10 (m, 1H), 3.09-3.03 (m, 1H), 2.55 (s, 3H), 2.01-1.95 (m, 2H), 1.93-1.88 (m, 1H), 1.76-1.72 (m, 1H), 1.34 (d,J= 6.5 Hz, 3H). ESI-MS m / z 682.0 [M+H + ]

[0376]

[0377] Compound 21. 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-methylacetamide hydrochloride

[0378] Step 1) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-(4-(2-(methylamino)-2-oxoethoxy)phenyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (46)

[0379] To a solution of 41 (25 mg, 0.034 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added methylamine hydrochloride (2.8 mg, 0.051 mmol, 1.5 eq), HATU (19 mg, 0.051 mmol, 1.5 eq), and DIEA (12 μL, 0.068 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 46 (6.3 mg, 0.009 mmol, 25%) as a yellow solid.

[0380] 1 H NMR (500 MHz, CDCl3) δ 7.99-7.90 (m, 3H), 7.81 (s, 1H), 7.00 (d,J= 8.8 Hz, 2H), 6.57 (d,J= 7.1 Hz, 1H), 4.64 (d,J= 10.7 Hz, 1H), 4.57 (d,J= 5.7 Hz, 4H), 4.22-4.16 (m, 1H), 3.99 (dd,J= 10.7, 4.3 Hz, 1H), 3.73-3.67 (m, 2H), 3.49-3.41 (m, 1H), 3.41-3.33 (m, 1H), 3.24-3.16 (m, 1H), 3.11-3.03 (m, 1H), 2.95 (d,J= 5.0 Hz, 3H), 2.56 (s, 3H), 2.07 (s, 2H), 1.95-1.79 (m, 3H), 1.67-1.63 (m, 1H), 1.48 (s, 9H), 1.22 (d,J= 6.3 Hz, 3H). ESI-MS m / z 738.5 [M+H + ]

[0381] Step 2) Preparation of 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-methylacetamide hydrochloride (Compound 21)

[0382] To a solution of 46 (6.3 mg, 0.009 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.05 mL, 0.180 mmol, 20 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 21 (1.8 mg, 0.003 mmol, 31%) as a yellow solid.

[0383] 1 H NMR (500 MHz, MeOD) δ 8.51 (d,J= 7.0 Hz, 1H), 8.41 (s, 1H), 7.92 (d,J= 8.7 Hz, 2H), 7.23 (d,J= 8.8 Hz, 2H), 6.97 (d,J= 7.0 Hz, 1H), 4.67 (s, 2H), 4.64 (s, 2H), 4.34-4.30 (m, 1H), 4.02-3.99 (m, 1H), 3.98-3.95 (m, 1H), 3.92-3.88 (m, 2H), 3.50-3.47 (m, 1H), 3.20-3.12 (m, 2H), 2.85 (s, 3H), 2.57 (s, 3H), 2.01-1.93 (m, 3H), 1.79-1.74 (m, 1H), 1.36 (d,J= 2.5 Hz, 3H). ESI-MS m / z 638.3 [M+H + ]

[0384]

[0385] Compound 22. 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetic acid hydrochloride

[0386] To a solution of 41 (8.0 mg, 0.011 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.03 mL, 0.110 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 22 (4.3 mg, 0.006 mmol, 59%) as a pale yellow solid.

[0387] 1 H NMR (500 MHz, MeOD) δ 8.53 (d,J= 6.9 Hz, 1H), 8.42 (s, 1H), 7.89 (d,J= 8.7 Hz, 2H), 7.18 (d,J= 8.7 Hz, 2H), 7.00 (d,J= 6.9 Hz, 1H), 4.81 (s, 2H), 4.67 (s, 2H), 4.37-4.31 (m, 1H), 4.03-3.96 (m, 2H), 3.94-3.88 (m, 2H), 3.50 (d,J= 3.6 Hz, 1H), 3.24-3.17 (m, 1H), 3.17-3.10 (m, 1H), 2.58 (s, 3H), 2.03-1.92 (m, 3H), 1.79-1.74 (m, 1H), 1.35 (d,J= 6.4 Hz, 3H). ESI-MS m / z 625.1 [M+H + ]

[0388]

[0389] Compound 23. (S)-2-(3-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetic acid hydrochloride

[0390] Step 1) Preparation of methyl 2-(3-acetylphenoxy)acetate (47)

[0391] To a solution of 1-(3-hydroxyphenyl)ethanone (1.00 g, 7.35 mmol, 1.0 eq) in acetone (20 mL) at room temperature were added methyl bromoacetate (0.83 mL, 8.81 mmol, 1.2 eq) and K2CO3 (1.20 g, 8.81 mmol, 1.2 eq). The reaction mixture was stirred at 80 °C for 3 h. The reaction was quenched with water and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (7 / 3) as an eluent to give 47 (1.52 g, 7.30 mmol, 99%) as a clear oil.

[0392] Step 2) Preparation of methyl 2-(3-(2-bromoacetyl)phenoxy)acetate (48)

[0393] To a solution of 47 (500 mg, 2.401 mmol, 1.0 eq) in chloroform (10 mL) at room temperature, N-bromosuccinimide (513 mg, 2.882 mmol, 1.2 eq) and pTSAㆍH2O (91.3 mg, 0.480 mmol, 0.2 eq) were added. The reaction mixture was stirred at 70 °C for 5 h. The reaction was washed with NaHCO 3(aq), quenched with water and extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4 and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (7 / 3) as eluent to give 48 (578 mg, 2.01 mmol, 84%) as a white solid.

[0394] Step 3) Preparation of methyl 2-(3-(7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetate (49)

[0395] To a solution of 8 (150 mg, 0.272 mmol, 1.0 eq) in ethanol (2.0 mL) at room temperature were added 48 (82 mg, 0.286 mmol, 1.2 eq) and KI (9.0 mg, 0.054 mmol, 0.2 eq). The reaction mixture was stirred at 60 °C for 16 h. The reaction mass was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using Hex / EtOAc (1 / 9) as an eluent to give 49 (167 mg, 0.226 mmol, 80%) as a tan solid.

[0396] 1H NMR (500 MHz, MeOD) δ 8.26-8.20 (m, 2H), 7.60 (t,J= 1.9 Hz, 1H), 7.57 (d,J= 7.7 Hz, 1H), 7.38 (t,J= 8.0 Hz, 1H), 6.98-6.94 (m, 2H), 6.53 (d,J= 7.1 Hz, 1H), 4.82 (d,J= 9.8 Hz, 2H), 4.60 (s, 2H), 4.32-4.21 (m, 2H), 3.99 (dd,J= 10.7, 4.7 Hz, 1H), 3.85-3.73 (m, 4H), 3.69 (d,J= 8.7 Hz, 1H), 3.61-3.51 (m, 1H), 3.43 (td,J= 9.7, 8.7, 5.7 Hz, 2H), 3.30-3.26 (m, 1H), 2.53 (s, 3H), 1.94-1.79 (m, 3H), 1.73-1.66 (m, 1H), 1.49 (s, 9H), 1.17 (d,J= 6.3 Hz, 3H).

[0397] Step 4) Preparation of 2-(3-(7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetic acid (50)

[0398] To a solution of 49 (168 mg, 0.22 mmol, 1.0 eq) in MeOH (2.0 mL) at room temperature was added 2N-LiOH (0.44 mL, 0.88 mmol, 4.0 eq) in H2O. The reaction mixture was stirred at room temperature for 5 h. After TLC analysis showed the complete disappearance of the starting material, the reaction mixture was concentrated to remove THF and methanol. The reaction mass was purified by adding 1N-HCl (aq) The pH was adjusted to 2 using and extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed under vacuum to obtain 50 (119 mg, 0.16 mmol, 75%) as a yellow solid.

[0399] 1 H NMR (500 MHz, MeOD) δ 8.26-8.21 (m, 2H), 7.62-7.55 (m, 2H), 7.38 (t,J= 7.9 Hz, 1H), 7.00-6.94 (m, 2H), 6.53 (d,J= 7.2 Hz, 1H), 4.74 (s, 2H), 4.61 (s, 2H), 4.31-4.24 (m, 1H), 3.99 (dd,J= 10.7, 4.7 Hz, 1H), 3.76 (d,J= 8.7 Hz, 1H), 3.69 (d,J= 8.7 Hz, 1H), 3.62-3.55 (m, 1H), 3.50-3.41 (m, 2H), 3.28 (d,J= 9.1 Hz, 1H), 2.53 (s, 3H), 1.94-1.79 (m, 3H), 1.73-1.65 (m, 1H), 1.49 (s, 9H), 1.17 (d,J= 6.3 Hz, 3H). ESI-MS m / z 725.1 [M+H + ]

[0400] Step 5) Preparation of (S)-2-(3-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetic acid hydrochloride (Compound 23)

[0401] To a solution of 50 (14.9 mg, 0.02 mmol, 1.0 eq) in DCM (10 mL) at 0 °C was added 4N-HCl (0.06 mL, 0.24 mmol, 12 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 23 (9.5 mg, 0.014 mmol, 71%) as a yellow solid.

[0402] 1H NMR (500 MHz, MeOD) δ 8.57-8.51 (m, 2H), 7.55 (dd,J= 6.4, 3.1 Hz, 3H), 7.19 (dd,J= 6.2, 2.9 Hz, 1H), 7.01 (d,J= 7.2 Hz, 1H), 4.83 (s, 2H), 4.67 (s, 2H), 4.37-4.31 (m, 1H), 4.00 (d,J= 9.0 Hz, 1H), 3.95 (d,J= 22.8 Hz, 1H), 3.90 (d,J= 9.0 Hz, 1H), 3.49 (d,J= 4.0 Hz, 1H), 3.26-3.07 (m, 3H), 2.58 (s, 3H), 2.05-1.91 (m, 3H), 1.77 (d,J= 13.0 Hz, 1H), 1.35 (d,J= 6.5 Hz, 3H). ESI-MS m / z 625.0 [M+H + ]

[0403]

[0404] Compound 24. 2-(3-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(piperazin-1-yl)ethan-1-one dihydrochloride

[0405] Step 1) Preparation of tert-butyl 4-(2-(3-(7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetyl)piperazine-1-carboxylate (51)

[0406] To a solution of 50 (40 mg, 0.055 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 4-amino-1-Boc-piperidine (11 mg, 0.061 mmol, 1.1 eq), HATU (23 mg, 0.061 mmol, 1.1 eq), and DIEA (29 μL, 0.165 mmol, 3.0 eq). The reaction mixture was stirred at room temperature for 6 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (1 / 9) as the eluent to give 51 (37.7 mg, 0.042 mmol, 76%) as a yellow solid.

[0407] 1 H NMR (500 MHz, MeOD) δ 8.25-8.19 (m, 2H), 7.63 (d,J= 2.5 Hz, 1H), 7.55 (d,J= 7.7 Hz, 1H), 7.37 (t,J= 8.0 Hz, 1H), 7.00 (dd,J= 8.3, 2.7 Hz, 1H), 6.95 (d,J= 10.7 Hz, 1H), 6.52 (d,J= 7.2 Hz, 1H), 4.94 (s, 2H), 4.60 (s, 2H), 4.30-4.24 (m, 1H), 3.99 (dd,J= 10.7, 4.6 Hz, 1H), 3.76 (d,J= 8.9 Hz, 1H), 3.68 (d,J= 8.7 Hz, 1H), 3.64 (t,J= 4.7 Hz, 4H), 3.58 (q,J= 5.2 Hz, 3H), 3.48 (d,J= 12.2 Hz, 3H), 3.45-3.42 (m, 2H), 3.31-3.23 (m, 1H), 2.53 (s, 3H), 1.93-1.78 (m, 3H), 1.72-1.64 (m, 1H), 1.49 (d,J= 4.5 Hz, 18H), 1.17 (d,J= 6.3 Hz, 3H). ESI-MS m / z 893.2 [M+H + ]

[0408] Step 2) Preparation of 2-(3-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(piperazin-1-yl)ethan-1-one dihydrochloride (Compound 24)

[0409] To a solution of 51 (37 mg, 0.041 mmol, 1.0 eq) in DCM (10 mL) at 0 °C was added 4N-HCl (0.10 mL, 0.41 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 24 (18.3 mg, 0.024 mmol, 57%) as a yellow solid.

[0410] 1 H NMR (500 MHz, MeOD) δ 8.55 (d,J= 7.9 Hz, 2H), 7.63 (t,J= 2.0 Hz, 1H), 7.60-7.52 (m, 2H), 7.23-7.19 (m, 1H), 6.99 (d,J= 7.2 Hz, 1H), 5.06 (s, 2H), 4.67 (s, 2H), 4.33 (dd,J= 6.6, 4.2 Hz, 1H), 4.00 (d,J= 9.0 Hz, 1H), 3.97 (s, 1H), 3.92-3.86 (m, 5H), 3.49 (d,J= 4.0 Hz, 1H), 3.38 (d,J= 6.8 Hz, 2H), 3.21 (d,J= 11.2 Hz, 1H), 3.13 (t,J= 11.9 Hz, 1H), 2.58 (s, 3H), 2.02-1.91 (m, 3H), 1.77 (d,J= 13.0 Hz, 1H), 1.35 (d,J=6.5 Hz, 3H). ESI-MS m / z 693.1 [M+H + ]

[0411]

[0412] Compound 25. 2-(3-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(2-methoxyethyl)acetamide hydrochloride

[0413] Step 1) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-(3-(2-((2-methoxyethyl)amino)-2-oxoethoxy)phenyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (52)

[0414] To a solution of 50 (40 mg, 0.055 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 2-methoxyethylamine (5.3 mg, 0.061 mmol, 1.1 eq), HATU (23 mg, 0.061 mmol, 1.1 eq), and DIEA (29 μL, 0.165 mmol, 3.0 eq). The reaction mixture was stirred at room temperature for 6 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (1 / 9) as an eluent to give 52 (29.2 mg, 0.037 mmol, 67%) as a yellow solid.

[0415] 1H NMR (500 MHz, MeOD) δ 8.24-8.19 (m, 2H), 7.65-7.62 (m, 1H), 7.58 (d,J= 7.8 Hz, 1H), 7.39 (t,J= 7.9 Hz, 1H), 7.01 (dd,J= 8.2, 2.6 Hz, 1H), 6.95 (d,J= 10.7 Hz, 1H), 6.51 (d,J= 7.1 Hz, 1H), 4.63 (s, 2H), 4.60 (s, 2H), 4.30-4.21 (m, 1H), 3.99 (dd,J= 10.7, 4.7 Hz, 1H), 3.76 (d,J= 8.7 Hz, 1H), 3.68 (d,J= 8.7 Hz, 1H), 3.57 (ddd,J= 10.9, 6.9, 3.4 Hz, 1H), 3.51 (d,J= 3.2 Hz, 4H), 3.46-3.39 (m, 2H), 3.35 (s, 3H), 3.32-3.24 (m, 1H), 2.52 (s, 3H), 1.89-1.82 (m, 3H), 1.68 (ddd,J= 12.8, 7.5, 3.2 Hz, 1H), 1.48 (s, 9H), 1.17 (d,J= 6.3 Hz, 3H). ESI-MS m / z 782.6 [M+H + ]

[0416] Step 2) Preparation of 2-(3-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(2-methoxyethyl)acetamide hydrochloride (Compound 25)

[0417] To a solution of 52 (29 mg, 0.037 mmol, 1.0 eq) in DCM (10 mL) at 0 °C was added 4N-HCl (0.09 mL, 0.37 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 25 (17.7 mg, 0.025 mmol, 66%) as a yellow solid.

[0418] 1 H NMR (500 MHz, MeOD) δ 8.56-8.52 (m, 2H), 7.58 (d,J= 2.3 Hz, 2H), 7.56 (d,J= 8.1 Hz, 1H), 7.26-7.23 (m, 1H), 7.01 (d,J= 7.2 Hz, 1H), 4.68 (d,J= 3.1 Hz, 3H), 4.33 (dd,J= 6.5, 4.2 Hz, 1H), 4.00 (d,J= 9.0 Hz, 1H), 3.95 (d,J= 22.6 Hz, 1H), 3.90 (d,J= 8.9 Hz, 1H), 3.54-3.48 (m, 5H), 3.36 (d,J= 3.9 Hz, 3H), 3.22 (d,J= 11.0 Hz, 1H), 3.14 (t,J= 11.5 Hz, 1H), 2.58 (s, 3H), 2.02-1.91 (m, 3H), 1.77 (d,J= 12.9 Hz, 1H), 1.35 (d,J= 6.5 Hz, 3H). ESI-MS m / z 682.0 [M+H + ]

[0419]

[0420] Compound 26. 4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)benzoic acid hydrochloride

[0421] Step 1) Preparation of ethyl 4-(2-bromoacetyl)benzoate (53)

[0422] To a solution of ethyl 4-acetylbenzoate (450 mg, 2.34 mmol, 1.0 eq) in chloroform (10 mL) at room temperature were added N-bromosuccinimide (500 mg, 2.81 mmol, 1.2 eq) and pTsOHㆍH2O (89 mg, 0.47 mmol, 0.2 eq). The reaction mixture was stirred at 70 °C for 16 h. The reaction was washed with NaHCO 3(aq) , quenched with water and extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4 and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (1 / 4) as eluent to give 53 (500 mg, 1.84 mmol, 79%) as a white solid.

[0423] 1 H NMR (400 MHz, CDCl3) δ 8.20-8.17 (m, 2H), 8.09-8.02 (m, 2H), 4.49 (s, 2H), 4.44 (q,J= 7.1 Hz, 2H), 1.44 (t,J= 7.1 Hz, 3H). ESI-MS m / z 270.0 [M+H + ]

[0424] Step 2) Preparation of ethyl 4-(7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)benzoate (54)

[0425] To a solution of 8 (390 mg, 0.71 mmol, 1.0 eq) in ethanol (5.0 mL) at room temperature were added 53 (230 mg, 0.85 mmol, 1.2 eq) and KI (24 mg, 0.14 mmol, 0.2 eq). The reaction mixture was stirred at 65 °C for 16 h, and the reaction mass was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using DCM / MeOH (19 / 1) as an eluent to give 54 (130 mg, 0.18 mmol, 25%) as a pale yellow solid.

[0426] 1 H NMR (500 MHz, MeOD) δ 8.35 (s, 1H), 8.27 (d,J= 7.1 Hz, 1H), 8.14-8.07 (m, 4H), 6.55 (d,J= 7.1 Hz, 1H), 4.61 (s, 2H), 4.41 (q,J= 7.1 Hz, 2H), 4.30-4.25 (m, 1H), 4.01-3.97 (m, 1H), 3.76 (d,J= 8.7 Hz, 1H), 3.69 (d,J= 8.7 Hz, 1H), 3.62-3.55 (m, 1H), 3.49-3.41 (m, 2H), 3.28 (s, 1H), 2.54 (s, 3H), 1.94-1.82 (m, 3H), 1.71-1.66 (m, 1H), 1.49 (s, 9H), 1.43 (t,J= 7.1 Hz, 3H), 1.17 (d,J= 6.2 Hz, 3H). ESI-MS m / z 723.1 [M+H + ]

[0427] Step 3) Preparation of 4-(7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)benzoic acid (55)

[0428] To a solution of 54 (130 mg, 0.18 mmol, 1.0 eq) in THF (3.6 mL) and MeOH (0.9 mL) at room temperature was added 1N-LiOH (0.9 mL, 0.90 mmol, 5.0 eq) dissolved in H2O. The reaction mixture was stirred at room temperature for 1 h. After TLC analysis showed the complete disappearance of the starting material, the reaction mixture was concentrated to remove THF and methanol. The residue was purified by filtration with 1N-HCl (aq) The pH was adjusted to 2 using and extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo to give 55 (100 mg, 0.14 mmol, 94%) as a pale yellow solid.

[0429] 1 H NMR (500 MHz, MeOD) δ 8.34 (s, 1H), 8.27 (d,J= 7.2 Hz, 1H), 8.10 (q,J= 8.4 Hz, 4H), 6.54 (d,J= 7.1 Hz, 1H), 4.61 (s, 2H), 4.30-4.25 (m, 1H), 4.02-3.97 (m, 1H), 3.76 (d,J= 8.6 Hz, 1H), 3.69 (d,J= 8.7 Hz, 1H), 3.61-3.55 (m, 1H), 3.49-3.39 (m, 2H), 3.31-3.26 (m, 1H), 2.54 (s, 3H), 1.93-1.80 (m, 3H), 1.73-1.66 (m, 1H), 1.49 (s, 9H), 1.17 (d,J= 6.3 Hz, 3H). ESI-MS m / z 695.0 [M+H + ]

[0430] Step 4) Preparation of 4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)benzoic acid hydrochloride (Compound 26)

[0431] To a solution of 55 (10 mg, 0.014 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.04 mL, 0.140 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 26 (6.5 mg, 0.010 mmol, 71%) as a pale yellow solid.

[0432] 1 H NMR (500 MHz, MeOD) δ 8.35 (s, 1H), 8.27 (d,J= 7.1 Hz, 1H), 8.14-8.07 (m, 4H), 6.55 (d,J= 7.1 Hz, 1H), 4.61 (s, 2H), 4.41 (q,J= 7.1 Hz, 2H), 4.30-4.25 (m, 1H), 4.01-3.97 (m, 1H), 3.76 (d,J= 8.7 Hz, 1H), 3.69 (d,J= 8.7 Hz, 1H), 3.62-3.55 (m, 1H), 3.49-3.41 (m, 2H), 3.31-3.27 (m, 1H), 2.54 (s, 3H), 1.94-1.82 (m, 3H), 1.71-1.66 (m, 1H), 1.49 (s, 9H), 1.43 (t,J= 7.1 Hz, 3H), 1.17 (d,J= 6.2 Hz, 3H). ESI-MS m / z 595.0 [M+H + ]

[0433]

[0434] Compound 27. (4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenyl)(piperazin-1-yl)methanone dihydrochloride

[0435] Step 1) Preparation of tert-butyl 4-(4-(7-((5-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)benzoyl)piperazine-1-carboxylate (56)

[0436] To a solution of 55 (20 mg, 0.029 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 1-Boc-piperazine (8.1 mg, 0.044 mmol, 1.5 eq), HATU (17 mg, 0.044 mmol, 1.5 eq), and DIEA (10 μL, 0.058 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 56 (15 mg, 0.017 mmol, 60%) as a white solid.

[0437] 1 H NMR (500 MHz, CDCl3) δ 8.24 (d,J= 8.1 Hz, 1H), 8.13-8.00 (m, 4H), 7.58-7.46 (m, 3H), 6.63 (d,J= 7.2 Hz, 1H), 4.68-4.62 (m, 3H), 4.22-4.18 (m, 1H), 4.04-3.99 (m, 1H), 3.73-3.71 (m, 2H), 3.55-3.43 (m, 8H), 3.32-3.25 (m, 1H), 3.19-3.13 (m, 1H), 2.59-2.55 (m, 3H), 1.97-1.83 (m, 4H), 1.75-1.69 (m, 1H), 1.49 (d,J= 8.6 Hz, 18H), 1.23 (d,J= 6.2 Hz, 3H). ESI-MS m / z 863.1 [M+H + ]

[0438] Step 2) Preparation of (4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenyl)(piperazin-1-yl)methanone dihydrochloride (Compound 27)

[0439] To a solution of 56 (15 mg, 0.017 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.04 mL, 0.170 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h. The solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 27 (12.0 mg, 0.016 mmol, 94%) as a pale yellow solid.

[0440] 1 H NMR (500 MHz, MeOD) δ 8.62 (s, 1H), 8.57 (d,J= 7.2 Hz, 1H), 8.10 (d,J= 8.3 Hz, 2H), 7.75 (d,J= 8.3 Hz, 2H), 7.00 (d,J= 7.2 Hz, 1H), 4.68 (s, 2H), 4.36-4.31 (m, 1H), 4.05-3.86 (m, 8H), 3.50 (d,J= 4.1 Hz, 1H), 3.36 (s, 4H), 3.24-3.18 (m, 1H), 3.18-3.11 (m, 1H), 2.58 (s, 3H), 2.03-1.98 (m, 2H), 1.96-1.90 (m, 1H), 1.80-1.73 (m, 1H), 1.35 (d,J= 6.5 Hz, 3H). ESI-MS m / z 663.1 [M+H + ]

[0441]

[0442] Compound 28. 4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-N-(2-methoxyethyl)benzamide hydrochloride

[0443] Step 1) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-(4-((2-methoxyethyl)carbamoyl)phenyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (57)

[0444] To a solution of 55 (20 mg, 0.029 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 2-methoxyethylamine (3.8 μL, 0.044 mmol, 1.5 eq), HATU (17 mg, 0.044 mmol, 1.5 eq), and DIEA (10 μL, 0.058 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 57 (10 mg, 0.013 mmol, 46%) as a white solid.

[0445] 1H NMR (500 MHz, CDCl3) δ 8.14-8.04 (m, 2H), 8.03 (d,J= 8.1 Hz, 2H), 7.85 (d,J= 8.2 Hz, 2H), 6.89-6.72 (m, 1H), 6.56 (d,J= 7.0 Hz, 1H), 4.70-4.59 (m, 3H), 4.21-4.17 (m, 1H), 4.01 (dd,J= 10.7, 4.3 Hz, 1H), 3.78-3.64 (m, 5H), 3.62 (t,J= 5.0 Hz, 2H), 3.54-3.47 (m, 1H), 3.42-3.37 (m, 1H), 3.30-3.22 (m, 1H), 3.16-3.06 (m, 1H), 2.56 (s, 3H), 1.97-1.81 (m, 4H), 1.72-1.65 (m, 1H), 1.56-1.41 (m, 11H), 1.23 (d,J= 6.2 Hz, 3H). ESI-MS m / z 752.0 [M+H + ]

[0446] Step 2) Preparation of 4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-N-(2-methoxyethyl)benzamide hydrochloride (Compound 28)

[0447] To a solution of 57 (10 mg, 0.013 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.03 mL, 0.130 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 28 (7.9 mg, 0.011 mmol, 86%) as a pale yellow solid.

[0448] 1H NMR (500 MHz, MeOD) δ 8.61 (s, 1H), 8.57 (d,J= 7.2 Hz, 1H), 8.06 (s, 4H), 7.01 (d,J= 7.2 Hz, 1H), 4.68 (s, 2H), 4.37-4.30 (m, 1H), 4.01 (d,J= 9.0 Hz, 1H), 3.99-3.93 (m, 1H), 3.90 (d,J= 9.0 Hz, 1H), 3.62 (s, 5H), 3.50 (d,J= 4.0 Hz, 1H), 3.42 (s, 3H), 3.25-3.17 (m, 1H), 3.17-3.10 (m, 1H), 2.58 (s, 3H), 2.03-1.97 (m, 2H), 1.97-1.90 (m, 1H), 1.80-1.74 (m, 1H), 1.35 (d,J= 6.5 Hz, 3H). ESI-MS m / z 652.0 [M+H + ]

[0449]

[0450] Compound 29. (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((2-(4-aminophenyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride

[0451] Step 1) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-(4-nitrophenyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (58)

[0452] To a solution of 8 (170 mg, 0.031 mmol, 1.0 eq) in ethanol (10 mL) at room temperature were added 2-bromo-4-nitroacetophenone (9.1 mg, 0.037 mmol, 1.2 eq) and KI (1.0 mg, 0.006 mmol, 0.2 eq). The reaction mixture was stirred at 65 °C for 16 h, and the reaction mass was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using DCM / MeOH (0.5 / 9.5) as the eluent to give 58 (80 mg, 0.11 mmol, 37%) as a yellow solid.

[0453] ESI-MS m / z 696.0 [M+H + ]

[0454] Step 2) Preparation of tert-butyl ((3S,4S)-8-(5-((2-(4-aminophenyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (59)

[0455] To a solution of 58 (80 mg, 0.11 mmol, 1.0 eq) in ethanol (2.0 mL) and H2O (2.0 mL) at room temperature were added Fe (31 mg, 0.55 mmol, 5.0 eq) and NH4Cl (29 mg, 0.55 mmol, 5.0 eq). The reaction mixture was stirred at 65 °C for 2 h. The reaction mixture was cooled to room temperature, diluted with ethyl acetate, and filtered through a silica pad. The filtrate was washed with brine, the organic layer was separated, dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 59 (56 mg, 0.084 mmol, 62%) as a yellow solid.

[0456] 1H NMR (400 MHz, CDCl3) δ 7.93 (d,J= 7.0 Hz, 1H), 7.80 (d,J= 8.3 Hz, 2H), 7.75 (s, 1H), 6.76 (d,J= 8.4 Hz, 2H), 6.55 (d,J= 7.0 Hz, 1H), 4.64 (d,J= 10.9 Hz, 1H), 4.57 (s, 2H), 4.22-4.17 (m, 1H), 3.99 (dd,J= 10.7, 4.3 Hz, 1H), 3.73-3.68 (m, 2H), 3.47-3.40 (m, 1H), 3.35 (s, 1H), 3.22-3.15 (m, 1H), 3.10-3.02 (m, 1H), 2.56 (s, 3H), 1.95-1.80 (m, 5H), 1.68-1.62 (m, 2H), 1.48 (s, 8H), 1.22 (d,J= 6.3 Hz, 3H). ESI-MS m / z 666.0 [M+H + ]

[0457] Step 3) Preparation of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((2-(4-aminophenyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 29)

[0458] To a solution of 59 (4.0 mg, 0.006 mmol, 1.0 eq) in DCM (3.0 mL) at 0 °C was added 4N-HCl (0.02 mL, 0.060 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 29 (3.6 mg, 0.006 mmol, 100%) as a pale yellow solid.

[0459] 1H NMR (400 MHz, MeOD) δ 8.59-8.50 (m, 2H), 8.03 (d,J= 8.3 Hz, 2H), 7.46 (d,J= 8.2 Hz, 2H), 7.00-6.92 (m, 1H), 4.67 (s, 2H), 4.36-4.29 (m, 1H), 4.00 (d,J= 8.8 Hz, 1H), 3.98-3.92 (m, 1H), 3.90 (d,J= 9.0 Hz, 1H), 3.50 (d,J= 3.4 Hz, 1H), 3.24-3.17 (m, 1H), 3.16-3.09 (m, 1H), 2.57 (s, 3H), 2.10-1.89 (m, 4H), 1.80-1.73 (m, 1H), 1.35 (d,J= 6.4 Hz, 3H). ESI-MS m / z 566.0 [M+H + ]

[0460]

[0461] Compound 30. N-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenyl)acetamide hydrochloride

[0462] Step 1) Preparation of tert-butyl ((3S,4S)-8-(5-((2-(4-acetamidophenyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (60)

[0463] To a solution of 59 (20 mg, 0.028 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added acetic anhydride (13 μL, 0.140 mmol, 5.0 eq). After stirring at room temperature for 1 h, the reaction was quenched with water and extracted with DCM (2 X 25 mL). The organic layer was separated, dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 60 (15 mg, 0.021 mmol, 71%) as a yellow solid.

[0464] 1 H NMR (400 MHz, MeOD) δ 8.29 (d,J= 7.2 Hz, 1H), 8.22 (s, 1H), 7.90 (d,J= 8.7 Hz, 2H), 7.70 (d,J= 8.7 Hz, 2H), 6.62 (d,J= 7.2 Hz, 1H), 4.61 (s, 2H), 4.30-4.22 (m, 1H), 4.03-3.96 (m, 1H), 3.76 (d,J= 8.7 Hz, 1H), 3.69 (d,J= 8.7 Hz, 1H), 3.63-3.55 (m, 1H), 3.52-3.41 (m, 2H), 3.32-3.26 (m, 1H), 2.53 (s, 3H), 2.17 (s, 3H), 1.93-1.80 (m, 3H), 1.72-1.63 (m, 1H), 1.49 (s, 9H), 1.17 (d,J= 6.3 Hz, 3H). ). ESI-MS m / z 708.0 [M+H + ]

[0465] Step 2) Preparation of N-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenyl)acetamide hydrochloride (Compound 30)

[0466] To a solution of 60 (11 mg, 0.016 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.04 mL, 0.160 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 30 (7.1 mg, 0.011 mmol, 71%) as a pale yellow solid.

[0467] 1 H NMR (400 MHz, MeOD) δ 8.50 (d,J= 7.4 Hz, 1H), 8.43 (s, 1H), 7.90 (d,J= 8.9 Hz, 2H), 7.82 (d,J= 8.8 Hz, 2H), 6.97 (d,J= 7.2 Hz, 1H), 4.66 (s, 2H), 4.36-4.31 (m, 1H), 4.02-3.88 (m, 4H), 3.50-3.47 (m, 1H), 3.17-3.12 (m, 1H), 2.58 (s, 3H), 2.19 (s, 3H), 2.07-1.94 (m, 4H), 1.79-1.73 (m, 1H), 1.35 (d,J= 6.3 Hz, 3H). ESI-MS m / z 608.1 [M+H + ]

[0468]

[0469] Compound 31. (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(4-methoxyphenyl))imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride

[0470] Step 1) Preparation of ethyl (S)-6-bromo-3-(1-((tert-butoxycarbonyl)amino)-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-5-methylpyrazine-2-carboxylate (61)

[0471] To a solution of 1 (3.00 g, 10.73 mmol, 1.0 eq) and (S)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine dihydrochloride (3.57 g, 12.88 mmol, 1.2 eq) in DMA (50 mL) at room temperature was added DIEA (11.2 mL, 64.38 mmol, 6.0 eq). The reaction mixture was stirred at 85 °C for 4 h and cooled to room temperature. Boc2O (9.40 g, 45.93 mmol, 4.0 eq) was added to the reaction solution, and the resulting brown solution was allowed to stand at room temperature for 16 h. The reaction was quenched with brine, water, and then extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (3 / 7) as the eluent to obtain 61 (4.40 g, 8.07 mmol, 75%) as a yellow solid.

[0472] 1 H NMR (400 MHz, CDCl3) δ 7.25 (p,J= 5.3 Hz, 4H), 5.00 (d,J= 10.0 Hz, 1H), 4.65 (d,J= 10.0 Hz, 1H), 4.42 (qd,J= 7.1, 2.4 Hz, 2H), 3.91-3.76 (m, 2H), 3.30-3.21 (m, 2H), 3.06 (d,J= 15.9 Hz, 1H), 2.81 (d,J= 15.7 Hz, 1H), 2.59-2.51 (m, 3H), 1.95 (ddd,J= 13.5, 11.3, 4.1 Hz, 1H), 1.78-1.72 (m, 2H), 1.53-1.46 (m, 9H), 1.44-1.38 (m, 3H).

[0473] Step 2) Preparation of tert-butyl(S)-(1'-(5-bromo-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (62)

[0474] To a solution of 61 (4.40 g, 8.07 mmol, 1.0 eq) in DCM (120 mL) at -78 °C was added dropwise 1 M DIBAL-H in hexane (33 mL, 32.27 mmol, 4.0 eq). The solution was then stirred at -78 °C for 0.5 h and then warmed to room temperature for 0.5 h. The reaction was quenched by pouring into saturated aqueous Rochelle salt solution. The resulting slurry was warmed to room temperature with vigorous stirring for 1 h, after which DCM and water were added. The organic layer was separated, and the aqueous layer was extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The mixture was purified by silica gel column chromatography using Hex / EtOAc (1 / 1) as the eluent to obtain 62 (2.61 g, 5.18 mmol, 67%) as a yellow solid.

[0475] 1 H NMR (400 MHz, DMSO) δ 7.22-7.18 (m, 4H), 5.45 (t,J= 5.8 Hz, 1H), 4.82 (d,J= 9.8 Hz, 1H), 4.43 (d,J= 5.5 Hz, 2H), 3.56 (s, 1H), 3.13-3.00 (m, 3H), 2.70 (d,J= 16.1 Hz, 1H), 2.45 (d,J= 4.9 Hz, 3H), 1.81-1.54 (m, 4H), 1.42 (s, 9H).

[0476] Step 3) Preparation of tert-butyl(S)-(1'-(5-((2-amino-3-chloropyridin-4-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (63)

[0477] To a solution of 62 (930 mg, 1.850 mmol, 1.00 eq) in 1,4-dioxane (10 mL) at room temperature were added 2-amino-3-chloro-pyridine-4-thio, sodium salt (381 mg, 2.030 mmol, 1.50 eq), Pd2(dba)3 (85 mg, 0.092 mmol, 0.05 eq), Xantphos (107 mg, 0.185 mmol, 0.10 eq), and DIEA (0.93 mL, 5.54 mmol, 3.00 eq). The mixture was purged with N2 and evacuated three times. The reaction mixture was stirred at 95 °C for 16 h. The reaction was quenched with water and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4 and the solvent was removed under vacuum. The residue was purified by silica gel column chromatography using DCM / MeOH (19 / 1) as the eluent to give 63 (630 mg, 1.080 mmol, 58%) as a pale orange solid.

[0478] 1 H NMR (500 MHz, MeOD) δ 7.62 (d,J= 5.5 Hz, 1H), 7.23 (d,J= 2.5 Hz, 4H), 5.91 (d,J= 5.6 Hz, 1H), 4.95-4.92 (m, 1H), 4.65 (s, 2H), 3.91-3.76 (m, 2H), 3.32-3.26 (m, 2H), 3.14 (d,J= 15.7 Hz, 1H), 2.83 (d,J= 15.8 Hz, 1H), 2.50 (s, 3H), 2.01-1.94 (m, 1H), 1.89-1.80 (m, 1H), 1.77-1.68 (m, 1H), 1.60-1.43 (m, 10H). ESI-MS m / z 583.0 [M+H + ]

[0479] Step 4) Preparation of (S)-tert-butyl (1'-(5-((8-chloro-2-(4-methoxyphenyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl))-6-methylpyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (64)

[0480] To a solution of 63 (30 mg, 0.051 mmol, 1.0 eq) in ethanol (10 mL) at room temperature were added 2-chloro-1-(4-methoxyphenyl)ethanone (11 mg, 0.061 mmol, 1.2 eq) and KI (1.7 mg, 0.010 mmol, 0.2 eq). The reaction mixture was stirred at 65 °C for 5 h, cooled to room temperature, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography using DCM / MeOH (0.5 / 9.5) as an eluent to give 64 (9.0 mg, 0.013 mmol, 25%) as a yellow solid.

[0481] 1 H NMR (500 MHz, CDCl3) δ 8.03 (d,J= 5.7 Hz, 1H), 7.96 (d,J= 8.2 Hz, 2H), 7.91-7.85 (m, 1H), 7.34-7.30 (m, 1H), 7.27-7.21 (m, 3H), 6.97 (d,J= 8.2 Hz, 2H), 6.59 (d,J= 6.8 Hz, 1H), 5.03 (d,J= 9.6 Hz, 1H), 4.69 (d,J= 9.5 Hz, 1H), 4.64 (s, 2H), 3.95-3.83 (m, 4H), 3.62-3.52 (m, 2H), 3.22-3.12 (m, 2H), 3.06 (d,J= 15.6 Hz, 1H), 2.82 (d,J= 15.9 Hz, 1H), 2.57 (s, 3H), 2.06-2.00 (m, 1H), 1.87-1.73 (m, 3H), 1.56-1.47 (m, 12H). ESI-MS m / z 713.6 [M+H + ]

[0482] Step 5) Preparation of (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(4-methoxyphenyl))imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 31)

[0483] To a solution of compound 64 (9.0 mg, 0.013 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.03 mL, 0.130 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 31 (5.0 mg, 0.008 mmol, 61%) as a pale yellow solid.

[0484] 1 H NMR (500 MHz, MeOD) δ 8.55 (d,J= 7.0 Hz, 1H), 8.42 (s, 1H), 7.89 (d,J= 8.5 Hz, 2H), 7.55 (d,J= 7.3 Hz, 1H), 7.46-7.41 (m, 2H), 7.40-7.34 (m, 1H), 7.17 (d,J= 8.5 Hz, 2H), 7.01 (d,J= 7.0 Hz, 1H), 4.69 (s, 2H), 4.47 (s, 1H), 4.05 (d,J= 13.8 Hz, 1H), 3.95-3.88 (m, 4H), 3.44-3.36 (m, 1H), 3.32-3.24 (m, 1H), 3.21 (s, 2H), 2.59 (s, 3H), 2.11-2.03 (m, 1H), 1.97-1.90 (m, 1H), 1.86-1.76 (m, 1H), 1.71-1.63 (m, 1H). ESI-MS m / z 613.0 [M+H + ]

[0485]

[0486] Compound 32. (S)-7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-N-(2-methoxyethyl)imidazo[1,2-a]pyridine-2-carboxamide hydrochloride

[0487] Step 1) Preparation of (S)-ethyl 7-((5-(1-((tert-butoxycarbonyl)amino)-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridine-2-carboxylate (65)

[0488] To a solution of 63 (60 mg, 0.10 mmol, 1.0 eq) in ethanol (5.0 mL) at room temperature were added ethyl 3-bromo-2-oxopropanoate (14 μL, 0.11 mmol, 1.1 eq) and KI (3.3 mg, 0.02 mmol, 0.2 eq). The reaction mixture was stirred at 65 °C for 16 h, and the reaction mass was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using DCM / MeOH (19 / 1) as an eluent to give 65 (25 mg, 0.039 mmol, 36%) as a pale yellow solid.

[0489] 1H NMR (500 MHz, MeOD) δ 8.47 (s, 1H), 8.30 (d,J= 7.2 Hz, 1H), 7.29-7.19 (m, 4H), 6.63 (d,J= 7.4 Hz, 1H), 4.96-4.92 (m, 1H), 4.63 (s, 2H), 4.44 (q, 2H), 3.88-3.73 (m, 2H), 3.31-3.26 (m, 1H), 3.13 (d,J= 15.8 Hz, 1H), 2.82 (d,J= 15.6 Hz, 1H), 2.54 (s, 3H), 2.10-1.94 (m, 2H), 1.90-1.82 (m, 1H), 1.77-1.69 (m, 1H), 1.56-1.46 (m, 10H), 1.43 (t,J= 7.0 Hz, 3H). ESI-MS m / z 679.1 [M+H + ]

[0490] Step 2) Preparation of (S)-7-((5-(1-((tert-butoxycarbonyl)amino)-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridine-2-carboxylic acid (66)

[0491] To a solution of 65 (25 mg, 0.039 mmol, 1.0 eq) in THF (0.64 mL) and MeOH (0.16 mL) at room temperature was added 1N-LiOH (0.16 mL, 0.156 mmol, 4.0 eq) dissolved in H2O. The reaction mixture was stirred at room temperature for 2 h. After the starting material completely disappeared according to TLC analysis, the reaction mixture was concentrated to remove THF and methanol. The product-containing aqueous layer was separated by adding 1N-HCl (aq) The pH was adjusted to 2 using and extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo to give 66 (20 mg, 0.031 mmol, 83%) as an ivory solid.

[0492] 1H NMR (400 MHz, MeOD) δ 8.24 (d,J= 7.2 Hz, 1H), 8.12 (s, 1H), 7.26-7.17 (m, 4H), 6.51 (d,J= 7.2 Hz, 1H), 4.94-4.92 (m, 1H), 4.61 (s, 2H), 3.87-3.68 (m, 2H), 3.28-3.23 (m, 1H), 3.12 (d, 1H), 2.81 (d,J= 15.8 Hz, 1H), 2.52 (s, 3H), 2.09-1.86 (m, 2H), 1.86-1.78 (m, 1H), 1.75-1.67 (m, 1H), 1.54-1.46 (m, 10H). ESI-MS m / z 651.0 [M+H + ]

[0493] Step 3) Preparation of (S)-tert-butyl (1'-(5-((8-chloro-2-((2-methoxyethyl)carbamoyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (67)

[0494] To a solution of 66 (20 mg, 0.031 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 2-methoxyethylamine (4.0 μL, 0.047 mmol, 1.5 eq), HATU (18 mg, 0.047 mmol, 1.5 eq), and DIEA (11 μL, 0.062 mmol, 2.0 eq). The reaction mixture was stirred at room temperature for 16 h, and the reaction was quenched with water and brine. The mixture was extracted with EtOAc (2 X 25 mL), and the combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 67 (6.5 mg, 0.009 mmol, 30%) as a light brown solid.

[0495] 1H NMR (500 MHz, CDCl3) δ 8.15 (s, 1H), 7.96 (d,J= 7.0 Hz, 1H), 7.91-7.77 (m, 1H), 7.34-7.29 (m, 1H), 7.27-7.21 (m, 3H), 6.58 (d,J= 6.8 Hz, 1H), 5.03 (d,J= 9.7 Hz, 1H), 4.70 (d,J= 9.9 Hz, 1H), 4.64 (s, 2H), 3.72-3.66 (m, 2H), 3.66-3.56 (m, 4H), 3.42 (s, 3H), 3.26-3.14 (m, 2H), 3.06 (d,J= 15.9 Hz, 1H), 2.82 (d,J= 15.7 Hz, 1H), 2.58 (s, 3H), 2.07-1.90 (m, 2H), 1.89-1.84 (m, 1H), 1.82-1.75 (m, 1H), 1.56-1.48 (m, 10H). ESI-MS m / z 708.1 [M+H + ]

[0496] Step 4) Preparation of (S)-7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-N-(2-methoxyethyl)imidazo[1,2-a]pyridine-2-carboxamide hydrochloride (Compound 32)

[0497] To a solution of 67 (6.5 mg, 0.009 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.02 mL, 0.090 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for several minutes, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 32 (3.5 mg, 0.005 mmol, 62%) as a pale yellow solid.

[0498] 1H NMR (500 MHz, MeOD) δ 8.42 (s, 1H), 7.53 (d,J= 7.4 Hz, 1H), 7.47-7.35 (m, 4H), 6.76 (d,J= 7.2 Hz, 1H), 4.65 (s, 2H), 4.01-3.93 (m, 1H), 3.89-3.81 (m, 1H), 3.67-3.59 (m, 4H), 3.42 (s, 3H), 3.31-3.15 (m, 4H), 2.56 (s, 3H), 2.03 (m, 2H), 1.94-1.87 (m, 1H), 1.82-1.76 (m, 1H), 1.70-1.64 (m, 1H). ESI-MS m / z 608.0 [M+H + ]

[0499]

[0500] Compound 33. (S)-2-(4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetic acid hydrochloride

[0501] Step 1) Preparation of methyl (S)-2-(4-(7-((5-(1-((tert-butoxycarbonyl)amino)-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetate (68)

[0502] To a solution of 63 (300 mg, 0.51 mmol, 1.0 eq) in ethanol (5.0 mL) at room temperature were added 39 (155 mg, 0.56 mmol, 1.1 eq) and KI (17 mg, 0.10 mmol, 0.2 eq). The reaction mixture was stirred at 60 °C for 7 h, and the reaction mass was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using Hex / EtOAc (1 / 9) as an eluent to give 68 (180 mg, 0.23 mmol, 45%) as a pale ivory solid.

[0503] 1 H NMR (400 MHz, DMSO-d6) δ 8.38-8.33 (m, 2H), 7.90 (d,J= 8.8 Hz, 2H), 7.21 (dd,J= 13.9, 10.1 Hz, 6H), 7.03 (d,J= 8.8 Hz, 2H), 6.40 (d,J= 7.1 Hz, 1H), 5.35 (t,J= 5.7 Hz, 1H), 4.84 (d,J= 12.5 Hz, 3H), 4.46 (d,J= 5.8 Hz, 2H), 3.72 (s, 5H), 3.23 (d,J= 11.7 Hz, 1H), 3.08 (d,J= 15.8 Hz, 1H), 2.74 (d,J= 15.9 Hz, 1H), 2.45 (s, 3H), 1.68 (dt,J= 38.6, 13.2 Hz, 4H), 1.42 (s, 9H).

[0504] Step 2) Preparation of (S)-2-(4-(7-((5-(1-((tert-butoxycarbonyl)amino)-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetic acid (69)

[0505] To a solution of 68 (180 mg, 0.23 mmol, 1.0 eq) in MeOH (2.0 mL) at room temperature was added 2N-LiOH (0.47 mL, 0.92 mmol, 4.0 eq) in H2O. The reaction mixture was stirred at room temperature for 5 h. After TLC analysis showed the complete disappearance of the starting material, the reaction mixture was concentrated under reduced pressure to remove THF and methanol. The product-containing aqueous layer was separated into 1N-HCl (aq) The pH was adjusted to 2 using and extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo to give 69 (120 mg, 0.16 mmol, 68%) as a pale orange solid.

[0506] 1H NMR (400 MHz, DMSO-d6) δ 8.39-8.33 (m, 2H), 7.91-7.85 (m, 2H), 7.28-7.17 (m, 5H), 6.98 (d,J= 8.8 Hz, 2H), 6.40 (d,J= 7.1 Hz, 1H), 5.36 (t,J= 5.8 Hz, 1H), 4.84 (d,J= 9.8 Hz, 1H), 4.63 (s, 2H), 4.45 (d,J= 5.5 Hz, 2H), 3.74 (t,J= 15.3 Hz, 3H), 3.08 (d,J= 15.8 Hz, 1H), 2.74 (d,J= 15.8 Hz, 1H), 2.45 (s, 3H), 1.80-1.58 (m, 4H), 1.42 (s, 9H). ESI-MS m / z 757.2 [M+H + ]

[0507] Step 3) Preparation of (S)-2-(4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetic acid hydrochloride (Compound 33)

[0508] To a solution of 69 (8.0 mg, 0.011 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.02 mL, 0.066 mmol, 6.0 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 33 (4.9 mg, 0.007 mmol, 54%) as a yellow solid.

[0509] 1H NMR (500 MHz, MeOD) δ 8.55 (d,J= 7.1 Hz, 1H), 8.43 (s, 1H), 7.90 (d,J= 8.5 Hz, 2H), 7.55 (d,J= 7.6 Hz, 1H), 7.43 (d,J= 7.0 Hz, 2H), 7.39-7.35 (m, 1H), 7.21-7.16 (m, 2H), 7.01 (d,J= 7.1 Hz, 1H), 4.82 (s, 2H), 4.68 (s, 2H), 4.47 (s, 1H), 4.05 (d,J= 13.7 Hz, 1H), 3.93 (d,J= 13.9 Hz, 1H), 3.44-3.35 (m, 2H), 3.21 (s, 2H), 2.59 (s, 3H), 2.09-2.03 (m, 1H), 1.95-1.91 (m, 1H), 1.81 (d,J= 12.8 Hz, 1H), 1.71-1.65 (m, 1H). ESI-MS m / z 657.0 [M+H + ]

[0510]

[0511] Compound 34. (S)-2-(4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(piperazin-1-yl)ethanone dihydrochloride

[0512] Step 1) Preparation of tert-butyl(S)-4-(2-(4-(7-((5-(1-((tert-butoxycarbonyl)amino)-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetyl)piperazine-1-carboxylate (70)

[0513] To a solution of 69 (25 mg, 0.033 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 1-Boc-piperazine (4.0 μL, 0.047 mmol, 1.4 eq), HATU (13.8 mg, 0.036 mmol, 1.1 eq), and DIEA (17 μL, 0.099 mmol, 3.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (1 / 9) as the eluent to give 70 (17.7 mg, 0.019 mmol, 58%) as a yellow solid.

[0514] ESI-MS m / z 925.1 [M+H + ]

[0515] Step 2) Preparation of (S)-2-(4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(piperazin-1-yl)ethanone dihydrochloride (Compound 34)

[0516] To a solution of 70 (7.0 mg, 0.007 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.06 mL, 0.24 mmol, 34 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 34 (3.5 mg, 0.004 mmol, 57%) as a yellow solid.

[0517] 1H NMR (500 MHz, MeOD) δ 8.53 (d,J= 7.3 Hz, 1H), 8.43 (d,J= 2.2 Hz, 1H), 7.91 (d,J= 8.2 Hz, 2H), 7.54 (d,J= 7.5 Hz, 1H), 7.43 (d,J= 7.7 Hz, 2H), 7.37 (t,J= 7.4 Hz, 1H), 7.24 (d,J= 8.4 Hz, 2H), 6.99 (d,J= 7.2 Hz, 1H), 5.04 (s, 2H), 4.69 (s, 2H), 4.46 (s, 1H), 4.03 (d,J=13.6 Hz, 1H), 3.90 (t,J= 5.3 Hz, 5H), 3.37 (d,J= 8.2 Hz, 4H), 3.21 (s, 2H), 2.59 (s, 3H), 2.07 (d,J= 12.0 Hz, 1H), 1.92 (t,J= 12.1 Hz, 1H), 1.81 (d,J= 13.0 Hz, 1H), 1.68 (d,J= 13.4 Hz, 1H). ESI-MS m / z 725.0 [M+H + ]

[0518]

[0519] Compound 35. (S)-2-(4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(4-aminopiperidin-1-yl)ethan-1-one dihydrochloride

[0520] Step 1) Preparation of tert-butyl(S)-(1'-(5-((2-(4-(2-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-2-oxoethoxy)phenyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (71)

[0521] To a solution of 69 (25 mg, 0.033 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 4-(tert-butoxycarbonylamino)piperidine (7.7 mg, 0.036 mmol, 1.5 eq), HATU (13.8 mg, 0.036 mmol, 1.5 eq), and DIEA (17 μL, 0.099 mmol, 3.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (1 / 9) as the eluent to give 71 (15.3 mg, 0.016 mmol, 49%) as a yellow solid.

[0522] 1 H NMR (500 MHz, MeOD) δ 8.24 (d,J= 7.1 Hz, 1H), 8.14 (s, 1H), 7.92 (d,J= 8.4 Hz, 2H), 7.23 (s, 4H), 7.08 (d,J= 8.6 Hz, 2H), 6.54 (d,J= 7.2 Hz, 1H), 4.96-4.90 (m, 3H) 4.61 (d,J= 8.7 Hz, 3H), 4.40 (d,J= 13.1 Hz, 1H), 3.98 (d,J= 14.1 Hz, 1H), 3.75 (dd,J= 25.1, 13.3 Hz, 2H), 3.63 (s, 2H), 3.26 (s, 2H), 3.14 (d,J= 15.4 Hz, 2H), 2.92 (t,J= 12.8 Hz, 1H), 2.82 (d,J= 15.7 Hz, 1H), 2.55 (s, 3H), 2.01-1.81 (m, 5H), 1.73 (d,J= 13.0 Hz, 1H), 1.51 (s, 9H), 1.46 (s, 9H). ESI-MS m / z 939.1 [M+H + ]

[0523] Step 2) Preparation of (S)-2-(4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(4-aminopiperidin-1-yl)ethan-1-one dihydrochloride (Compound 35)

[0524] To a solution of 71 (8.0 mg, 0.009 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.06 mL, 0.240 mmol, 27 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 h, and the solvent was concentrated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 35 (5.9 mg, 0.007 mmol, 77%) as a yellow solid.

[0525] 1H NMR (500 MHz, MeOD) δ 8.55 (d,J= 7.2 Hz, 1H), 8.44 (s, 1H), 7.91 (d,J= 8.4 Hz, 2H), 7.55 (d,J= 7.5 Hz, 1H), 7.43 (d,J= 7.3 Hz, 2H), 7.38 (d,J= 7.3 Hz, 1H), 7.22 (d,J= 8.4 Hz, 2H), 7.01 (d,J= 7.2 Hz, 1H), 5.02 (t,J= 13.7 Hz, 3H), 4.69 (s, 2H), 4.64 (d,J= 13.8 Hz, 1H), 4.46 (s, 1H), 4.16-4.01 (m, 2H), 3.93 (d,J= 13.8 Hz, 1H), 3.51-3.36 (m, 3H), 3.28 (d,J= 13.1 Hz, 1H), 3.21 (s, 2H), 2.84 (t,J= 12.9 Hz, 1H), 2.59 (s, 3H), 2.19-2.05 (m, 2H), 1.93 (t,J= 12.2 Hz, 1H), 1.81 (d,J= 12.9 Hz, 1H), 1.69 (t,J= 12.3 Hz, 2H), 1.60-1.55 (m, 1H). ESI-MS m / z 739.1 [M+H + ]

[0526]

[0527] Compound 36. (S)-2-(4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(piperidin-4-yl)acetamide dihydrochloride

[0528] Step 1) Preparation of tert-butyl(S)-4-(2-(4-(7-((5-(1-((tert-butoxycarbonyl)amino)-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetamido)piperidine-1-carboxylate (72)

[0529] To a solution of 69 (25 mg, 0.033 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added tert-Butyl 4-aminopiperidine-1-carboxylate (7.2 mg, 0.036 mmol, 1.5 eq), HATU (13.8 mg, 0.036 mmol, 1.5 eq), and DIEA (17 μL, 0.099 mmol, 3.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (1 / 9) as the eluent to give 72 (15.5 mg, 0.016 mmol, 50%) as a yellow solid.

[0530] ESI-MS m / z 939.1 [M+H + ]

[0531] Step 2) Preparation of (S)-2-(4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(piperidin-4-yl)acetamide dihydrochloride (Compound 36)

[0532] To a solution of 72 (10 mg, 0.011 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.06 mL, 0.240 mmol, 22 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 36 (4.6 mg, 0.005 mmol, 50%) as a yellow solid.

[0533] 1 H NMR (500 MHz, MeOD) δ 8.53 (d,J= 7.2 Hz, 1H), 8.43 (s, 1H), 7.93 (d,J= 8.4 Hz, 2H), 7.54 (d,J= 7.5 Hz, 1H), 7.43 (d,J= 7.7 Hz, 2H), 7.38 (d,J= 7.3 Hz, 1H), 7.25 (d,J= 8.6 Hz, 2H), 6.99 (d,J= 7.2 Hz, 1H), 4.68 (d,J= 8.6 Hz, 4H), 4.46 (s, 1H), 4.11-4.00 (m, 2H), 3.92 (d,J= 13.8 Hz, 1H), 3.47 (d,J= 12.6 Hz, 2H), 3.40 (d,J= 12.5 Hz, 1H), 3.24-3.10 (m, 5H), 2.59 (s, 3H), 2.15 (d,J= 14.1 Hz, 2H), 2.05 (d,J= 11.3 Hz, 1H), 1.87 (dd,J= 25.5, 12.8 Hz, 3H), 1.68 (d,J= 13.4 Hz, 1H). ESI-MS m / z 739.1 [M+H + ]

[0534]

[0535] Compound 37. (S)-2-(4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(2-methoxyethyl)acetamide hydrochloride

[0536] Step 1) Preparation of tert-butyl(S)-(1'-(5-((8-chloro-2-(4-(2-((2-methoxyethyl)amino)-2-oxoethoxy)phenyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (73)

[0537] To a solution of 69 (25 mg, 0.033 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 2-methoxyethan-1-amine (3.1 μL, 0.036 mmol, 1.5 eq), HATU (13.8 mg, 0.036 mmol, 1.5 eq), and DIEA (17 μL, 0.099 mmol, 3.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (1 / 9) as the eluent to give 73 (14.2 mg, 0.017 mmol, 53%) as a yellow solid.

[0538] ESI-MS m / z 814.0 [M+H + ]

[0539] Step 2) Preparation of (S)-2-(4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(2-methoxyethyl)acetamide hydrochloride (Compound 37)

[0540] To a solution of 73 (10 mg, 0.012 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.06 mL, 0.240 mmol, 20 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 h, and the solvent was evaporated in vacuo. The slurry was diluted by adding methanol and ethyl acetate. After stirring for 10 min, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 37 (2.4 mg, 0.003 mmol, 25%) as a yellow solid.

[0541] 1H NMR (500 MHz, MeOD) δ 8.53 (d,J= 7.1 Hz, 1H), 8.43 (s, 1H), 7.92 (d,J= 8.3 Hz, 2H), 7.55 (d,J= 7.5 Hz, 1H), 7.43 (d,J= 7.1 Hz, 2H), 7.37 (t,J= 7.3 Hz, 1H), 7.23 (d,J= 8.4 Hz, 2H), 6.99 (d,J= 7.1 Hz, 1H), 4.68 (d,J= 15.1 Hz, 4H), 4.46 (s, 1H), 4.04 (d,J= 13.8 Hz, 1H), 3.92 (d,J= 13.9 Hz, 1H), 3.51 (h,J= 4.9 Hz, 4H), 3.37 (d,J= 2.0 Hz, 5H), 3.21 (s, 2H), 2.59 (s, 3H), 2.06 (ddd,J= 14.8, 11.9, 3.8 Hz, 1H), 1.97-1.89 (m, 1H), 1.81 (d,J= 13.0 Hz, 1H), 1.67 (d,J= 13.3 Hz, 1H). ESI-MS m / z 714.0 [M+H + ]

[0542]

[0543] Compound 38. (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(4-methoxyphenyl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol

[0544] Step 1) Preparation of methyl 3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-bromopyrazine-2-carboxylate (74)

[0545] To a solution of methyl 3,6-dibromopyrazine-2-carboxylate (500 mg, 1.62 mmol, 1.0 eq) in MeCN (10 mL) at room temperature were added (3S,4S)-3-methyl-2-oxa-8-azaspiro[4,5]decane-4-amine dihydrochloride (332 mg, 1.95 mmol, 1.2 eq) and DIEA (0.70 mL, 4.07 mmol, 2.5 eq). The resulting mixture was stirred at room temperature for 12 h. The solvent was removed under reduced pressure, and the residue was partitioned between H2O / NH4OH (10:1) and EtOAc. The phases were separated, and the aqueous layer was extracted with EtOAc (2 X 25 mL). The combined organic phase was dried over anhydrous Na2SO4, filtered, and concentrated to obtain 74 (610 mg), which was used without further purification.

[0546] ESI-MS m / z 386.1 [M+H + ]

[0547] Step 2) Preparation of methyl 6-bromo-3-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)pyrazine-2-carboxylate (75)

[0548] To a solution of 74 (610 mg, 1.58 mmol, 1.0 eq) in DCM (3.0 mL) at room temperature were added Boc2O (0.54 mL, 2.29 mmol, 1.5 eq) and TEA (0.64 mL, 4.56 mmol, 3.0 eq). The mixture was stirred at room temperature for 8 h and concentrated under reduced pressure. The residue was purified by silica gel chromatography to give 75 (480 mg, 0.98 mmol, 65%) as a yellow solid.

[0549] 1H NMR (600 MHz, CDCl3) δ 8.45 (s, 1H), 7.67 (s, 1H), 4.47 (s, 1H), 3.83 (s, 3H), 3.78-3.69 (m, 4H), 3.43-3.35 (m, 4H), 3.14 (m, 2H), 1.50 (s, 3H), 1.4 (s, 9H). ESI-MS m / z 486.1 [M+H + ]

[0550] Step 3) Preparation of methyl 6-((2-amino-3-chloropyridin-4-yl)thio)-3-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)pyrazine-2-carboxylate (76)

[0551] To a solution of 75 (100 mg, 0.20 mmol, 1.00 eq) in 1,4-dioxane (5.0 mL) at room temperature were added 2-amino-3-chloro-pyridine-4-thio, sodium salt (37.52 mg, 0.20 mmol, 1.00 eq), Pd2(dba)3 (942 mg, 0.01 mmol, 0.05 eq), Xantphos (13 mg, 0.02 mmol, 0.10 eq), and DIEA (0.05 mL, 0.30 mmol, 1.5 eq). The mixture was purged with N2 and evacuated three times. The resulting mixture was stirred at 95 °C for 4 h. The reaction was quenched with water and extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (7 / 3) as the eluent to give 76 (63 mg, 0.11 mmol, 54%) as a yellow solid.

[0552] 1H NMR (600 MHz, CDCl3) δ 8.29 (d,J= 5.2 Hz, 1H), 7.60 (d,J= 6.5 Hz, 1H), 6.16 (d,J= 6.2 Hz, 1H), 4.65 (d,J= 10.7 Hz, 1H), 4.19-4.15 (m, 1H), 4.00-3.97 (m, 1H), 3.95 (s, 3H), 3.76-3.71 (m, 1H), 3.66 (td,J= 7.4, 3.4 Hz, 1H), 3.57-3.53 (m, 1H), 3.44-3.37 (m, 1H), 1.90-1.85 (m, 1H), 1.83-1.75 (m, 2H), 1.65 (d,J= 13.3, 3.7 Hz, 1H), 1.50 (s, 3H), 1.44 (d,J= 7.2 Hz, 9H). ESI-MS m / z 565.0 [M+H + ]

[0553] Step 4) Preparation of methyl 3-((3S,4S)-4-((tert-butoxycarbonyl)amino)-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(4-methoxyphenyl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazine-2-carboxylate (77)

[0554] To a solution of 76 (100 mg, 0.17 mmol, 1.0 eq) in ethanol (3.0 mL) at room temperature was added 2-bromo-1-(4-methoxyphenyl)ethanone (81 mg, 0.36 mmol, 2.1 eq). The reaction mixture was stirred at 85 °C for 18 h, and the progress of the reaction was monitored by TLC. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using Hex / EtOAc (3 / 7) as an eluent to give 77 (70 mg, 0.10 mmol, 59%) as a yellow solid.

[0555] 1H NMR (600 MHz, CDCl3) δ 8.29 (d,J= 5.2 Hz, 1H), 8.23 ​​(s, 1H), 7.89 (dd,J= 7.2 Hz, 2H), 7.60 (d,J= 6.5 Hz, 1H), 7.01 (dd,J= 7.2 Hz, 2H), 6.16 (d,J= 6.2 Hz, 1H), 4.65 (d,J= 10.7 Hz, 1H), 4.19-4.15 (m, 1H), 4.00-3.97 (m, 1H), 3.95 (s, 3H), 3.81 (s, 3H), 3.76-3.71 (m, 1H), 3.66 (td,J= 7.4, 3.4 Hz, 1H), 3.57-3.53 (m, 1H), 3.44-3.37 (m, 1H), 1.90-1.85 (m, 1H), 1.83-1.75 (m, 2H), 1.65 (dq,J= 13.3, 3.7 Hz, 1H), 1.44 (s, 9H), 1.12 (s, 3H). ESI-MS m / z 695.1 [M+H + ]

[0556] Step 5) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-(4-methoxyphenyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (78)

[0557] To a solution of 77 (100 mg, 0.14 mmol, 1.0 eq) in THF (5.0 mL) at 0 °C was added LiBH4 (6.2 mg, 0.28 mmol, 2.0 eq). The reaction mixture was stirred at 4 °C for 1 h and then quenched by the addition of water (1.0 mL) at 0 °C. The solution was filtered and concentrated to obtain a residue. The residue was purified by silica gel column chromatography using Hex / EtOAc (1 / 9) as an eluent to give 78 (60 mg, 0.09 mmol, 64%) as a yellow solid.

[0558] 1H NMR (600 MHz, CDCl3) δ 8.29 (d,J= 5.2 Hz, 1H), 8.23 ​​(s, 1H), 7.89 (dd,J= 7.2 Hz, 2H), 7.60 (d,J= 6.5 Hz, 1H), 7.01 (dd,J= 7.2 Hz, 2H), 6.16 (d,J= 6.2 Hz, 1H), 4.65 (d,J= 10.7 Hz, 1H), 4.20-3.97 (m, 4H), 3.95 (s, 3H), 3.76-3.71 (m, 1H), 3.66 (td,J= 7.4, 3.4 Hz, 1H), 3.57-3.53 (m, 1H), 3.44-3.37 (m, 1H), 1.90-1.85 (m, 1H), 1.83-1.75 (m, 2H), 1.65 (dq,J= 13.3, 3.7 Hz, 1H), 1.44 (s, 9H), 1.20 (s, 3H). ESI-MS m / z 667.1 [M+H + ]

[0559] Step 6) Preparation of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(4-methoxyphenyl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol (Compound 38)

[0560] To a solution of 78 (40 mg, 0.66 mmol, 1.0 eq) in DCM (3.0 mL) at 0 °C was added 4N-HCl (0.66 mL) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 16 h, and the solvent was removed under vacuum. The residue was purified by preparative HPLC to give compound 38 (22 mg, 0.038 mmol, 66%) as a yellow solid.

[0561] 1H NMR (600 MHz, CDCl3)δ8.29 (d,J= 5.2 Hz, 1H), 8.23 ​​(s, 1H), 7.89 (dd,J= 7.2 Hz, 2H), 7.60 (d,J= 6.5 Hz, 1H), 7.01 (dd,J= 7.2 Hz, 2H), 6.16 (d,J= 6.2 Hz, 1H), 4.65 (d,J= 10.7 Hz, 1H), 4.20-4.15 (m, 3H), 4.00-3.97 (m, 1H), 3.95 (s, 3H), 3.76-3.71 (m, 1H), 3.66 (td,J= 7.4, 3.4 Hz, 1H), 3.57-3.53 (m, 1H), 3.44-3.37 (m, 1H), 1.90-1.85 (m, 1H), 1.83-1.75 (m, 2H), 1.65 (dq,J= 13.3, 3.7 Hz, 1H), 1.50 (s, 3H). ESI-MS m / z 567.1. [M+H + ]

[0562]

[0563] Compound 39. (S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(4-methoxyphenyl))imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol

[0564] Step 1) Preparation of (S)-methyl 3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-bromopyrazine-2-carboxylate (79)

[0565] To a solution of methyl 3,6-dibromopyrazine-2-carboxylate (500 mg, 1.62 mmol, 1.0 eq) in MeCN (10 mL) at room temperature were added (S)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine dihydrochloride (409 mg, 2.02 mmol, 1.2 eq) and DIEA (0.44 mL, 2.53 mmol, 2.5 eq). The resulting mixture was stirred at room temperature for 12 h. The solvent was removed under reduced pressure, and the residue was partitioned between H2O / NH4OH (10:1) and EtOAc. The phases were separated, and the aqueous layer was extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated to obtain 79 (610 mg, 1.46 mmol), which was used without further purification.

[0566] ESI-MS m / z 417.1 [M+H + ]

[0567] Step 2) Preparation of (S)-methyl 6-bromo-3-(1-((tert-butoxycarbonyl)amino)-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)pyrazine-2-carboxylate (80)

[0568] To a solution of 79 (610 mg, 1.58 mmol, 1.0 eq) in DCM (3.0 mL) were added Boc2O (0.54 mL, 2.29 mmol, 1.5 eq) and TEA (0.64 mL, 4.56 mmol, 3.0 eq). The mixture was stirred at room temperature for 8 h and concentrated under reduced pressure. The residue was purified by silica gel chromatography to give 80 (530 mg, 1.02 mmol, 85%) as a yellow solid.

[0569] 1H NMR (600 MHz, CDCl3) δ8.45 (s, 1H), 7.67 (s, 1H), 7.23-7.21 (m, 4H), 4.47 (s, 1H), 3.83 (s, 3H), 3.78-3.69 (m, 4H), 3.43-3.35 (m, 4H), 3.17-3.12 (m, 2H), 1.40 (s, 9H). ESI-MS m / z 517.1 [M+H + ]

[0570] Step 3) Preparation of methyl (S)-6-((2-amino-3-chloropyridin-4-yl)thio)-3-(1-((tert-butoxycarbonyl)amino)-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)pyrazine-2-carboxylate (81)

[0571] To a solution of 80 (200 mg, 0.39 mmol, 1.00 eq) in 1,4-dioxane (5.0 mL) at room temperature were added 2-amino-3-chloro-pyridine-4-thio, sodium salt (70 mg, 0.39 mmol, 1.00 eq), Pd2(dba)3 (46 mg, 0.019 mmol, 0.05 eq), Xantphos (23 mg, 0.039 mmol, 0.10 eq), and DIEA (0.10 mL, 0.58 mmol, 1.5 eq). The mixture was purged with N2 and evacuated three times. After stirring at 95 °C for 16 h, the reaction was quenched with water and extracted with DCM (2 X 25 mL). The combined organic layers were dried over MaSO4 and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using Hex / EtOAc (7 / 3) as the eluent to give 81 (200 mg, 0.33 mmol, 86%) as a yellow solid.

[0572] 1H NMR (600 MHz, CDCl3) δ 8.32 (d,J= 17.9 Hz, 1H), 7.53 (d,J= 6.9 Hz, 1H), 7.25-7.22 (m, 4H), 6.26-6.21 (m, 1H), 4.97 (d,J= 9.6 Hz, 1H), 4.63 (d,J= 9.3 Hz, 1H), 4.00-3.97 (m, 1H), 3.95 (s, 3H), 3.94-3.90 (m, 1H), 3.41 (q,J= 10.4 Hz, 2H), 3.07-3.02 (m, 1H), 2.88-2.84 (m, 1H), 1.85-1.79 (m, 1H), 1.72 (d,J= 13.4 Hz, 2H), 1.47 (s, 9H). ESI-MS m / z 597.1 [M+H + ]

[0573] Step 4) Preparation of methyl (S)-3-(1-((tert-butoxycarbonyl)amino)-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(4-methoxyphenyl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazine-2-carboxylate (82)

[0574] To a solution of 81 (100 mg, 0.16 mmol, 1.0 eq) in ethanol (3.0 mL) at room temperature was added 2-bromo-1-(4-methoxyphenyl)ethanone (79 mg, 0.34 mmol, 2.1 eq). The reaction mixture was stirred at 85 °C for 18 h, and the progress of the reaction was monitored by TLC. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using Hex / EtOAc (3 / 7) as an eluent to give 82 (100 mg, 0.13 mmol, 86%) as a yellow solid.

[0575] 1H-NMR (600 MHz, CDCl3) δ 8.29 (d,J= 5.2 Hz, 1H), 8.23 ​​(s, 1H), 7.89 (dd,J= 7.2 Hz, 2H), 7.60 (d,J= 6.5 Hz, 1H), 7.25-7.22 (m, 4H), 7.01 (dd,J= 7.2 Hz, 2H), 6.16 (d,J= 6.2 Hz, 1H), 4.97 (d,J= 9.6 Hz, 1H), 4.63 (d,J= 9.3 Hz, 1H), 4.00-3.97 (m, 1H), 3.95 (s, 3H), 3.94 (s, 3H), 3.41 (q,J= 10.4 Hz, 2H), 3.07-3.02 (m, 1H), 2.88-2.84 (m, 1H), 1.94-1.89 (m, 1H), 1.85-1.79 (m, 1H), 1.72 (d,J= 13.4 Hz, 2H), 1.47 (s, 9H). ESI-MS m / z 727.1 [M+H + ]

[0576] Step 5) Preparation of tert-butyl(S)-(1'-(5-((8-chloro-2-(4-methoxyphenyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (83)

[0577] To a solution of 82 (100 mg, 0.13 mmol, 1.0 eq) in THF (5.0 mL) at 0 °C was added LiBH4 (5.8 mg, 0.26 mmol, 2.0 eq). The reaction mixture was stirred at 4 °C for 1 h and then quenched by the addition of water (1.0 mL) at 0 °C. The solution was filtered and concentrated to obtain a residue. The residue was purified by silica gel column chromatography using Hex / EtOAc (1 / 9) as an eluent to give 83 (60 mg, 0.08 mmol, 66%) as a yellow solid.

[0578] 1H-NMR (600 MHz, CDCl3) δ 8.29 (d,J= 5.2 Hz, 1H), 8.23 ​​(s, 1H), 7.89 (dd,J= 7.2 Hz, 2H), 7.60 (d,J= 6.5 Hz, 1H), 7.25-7.22 (m, 4H), 7.01 (dd,J= 7.2 Hz, 2H), 6.16 (d,J= 6.2 Hz, 1H), 4.97 (d,J= 9.6 Hz, 1H), 4.63 (d,J= 9.3 Hz, 1H), 4.00-3.97 (m, 1H), 3.95 (s, 3H), 3.92-3.90 (m, 2H), 3.41 (q,J= 10.4 Hz, 2H), 3.07-3.02 (m, 1H), 2.88-2.84 (m, 1H), 1.94-1.89 (m, 1H), 1.85-1.79 (m, 1H), 1.72 (d,J= 13.4 Hz, 2H), 1.47 (s, 9H). ESI-MS m / z 699.1 [M+H + ]

[0579] Step 6) Preparation of (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(4-methoxyphenyl))imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol (Compound 39)

[0580] To a solution of 83 (50 mg, 0.07 mmol, 1.0 eq) in DCM (5.0 mL) at 0 °C was added 4N-HCl (0.70 mL) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 16 h. The solvent was removed under vacuum, and the residue was purified by preparative HPLC to give compound 39 (25 mg, 0.04 mmol, 60%) as a yellow solid.

[0581] 1 H NMR (600 MHz, CDCl3)δ 1H-NMR (600 MHz, CDCl3) δ 8.29 (d,J= 5.2 Hz, 1H), 8.23 ​​(s, 1H), 7.89 (dd,J= 7.2 Hz, 2H), 7.60 (d,J= 6.5 Hz, 1H), 7.25-7.22 (m, 4H), 7.01 (dd,J= 7.2 Hz, 2H), 6.16 (d,J= 6.2 Hz, 1H), 4.97 (d,J= 9.6 Hz, 1H), 4.63 (d,J= 9.3 Hz, 1H), 4.00-3.97 (m, 1H), 3.95 (s, 3H), 3.94-3.91 (m, 2H), 3.41 (q,J= 10.4 Hz, 2H), 3.07-3.02 (m, 1H), 2.88-2.84 (m, 1H), 1.94-1.89 (m, 1H), 1.85-1.79 (m, 1H), 1.72 (d,J= 13.4 Hz, 2H). ESI-MS m / z 598.1 [M+H + ]

[0582]

[0583] Compound 40. (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(furan-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride

[0584] Step 1) Preparation of 2-bromo-1-(furan-2-yl)ethan-1-one (84)

[0585] To a solution of 1-(furan-2-yl)ethenone (300 mg, 2.72 mmol, 1.0 eq) in chloroform (10 mL) were added N-bromosuccinimide (NBS, 581 mg, 3.26 mmol, 1.2 eq) and pTsOHㆍH2O (103 mg, 0.54 mmol, 0.2 eq) at room temperature. The reaction mixture was stirred at 70 °C for 4 h. The reaction was quenched with saturated aqueous NaHCO3, water, and extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The crude mixture was purified by silica gel column chromatography using Hep / EtOAc (8 / 2) as an eluent to give 84 (177 mg, 1.15 mmol, 51%) as a white solid.

[0586] 1 H NMR (500 MHz, CDCl3) δ 7.69-7.64 (m, 1H), 7.36 (d,J= 3.6 Hz, 1H), 6.62 (dd,J= 3.6, 1.7 Hz, 1H), 4.34 (s, 2H). ESI-MS m / z 188.1 [M+H + ]

[0587] Step 2) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-(furan-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (85)

[0588] 84 (12 mg, 0.065 mmol, 1.2 eq) and KI (1.8 mg, 0.011 mmol, 0.2 eq) were added to a solution of 8 (30 mg, 0.054 mmol, 1.0 eq) in ethanol (2.0 mL) at room temperature. The reaction mixture was stirred at 65 °C for 16 h and concentrated under reduced pressure. The residue was then purified by silica gel column chromatography using DCM / MeOH (19 / 1) as the eluent to give 85 (10 mg, 0.016 mmol, 29%) as a white solid.

[0589] 1 H NMR (400 MHz, MeOD) δ 8.24 (d,J= 7.1 Hz, 1H), 8.07 (s, 1H), 7.60 (s, 1H), 7.03 (d,J= 3.1 Hz, 1H), 6.61-6.55 (m, 1H), 6.52 (d,J= 7.1 Hz, 1H), 4.60 (s, 2H), 4.32-4.22 (m, 1H), 4.06-3.96 (m, 1H), 3.76 (d,J= 8.6 Hz, 1H), 3.69 (d,J= 8.6 Hz, 1H), 3.62-3.38 (m, 3H), 3.31-3.22 (m, 1H), 2.53 (s, 3H), 1.96-1.78 (m, 3H), 1.71-1.62 (m, 1H), 1.49 (s, 9H), 1.17 (d,J= 6.2 Hz, 3H). ESI-MS m / z 642.1 [M+H + ]

[0590] Step 3) Preparation of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(furan-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (compound 40)

[0591] To a solution of 85 (10 mg, 0.016 mmol, 1.0 eq) in DCM (5.0 mL) at 0°C was added 4N-HCl (0.04 mL, 0.016 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted with methanol (0.3 mL) and ethyl acetate (1.5 mL). After stirring for several minutes, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to give compound 40 (6.0 mg, 0.010 mmol, 66%) as a pale yellow solid.

[0592] 1H NMR (400 MHz, MeOD) δ 8.50 (d,J= 7.2 Hz, 1H), 8.38 (s, 1H), 7.81 (s, 1H), 7.25 (d,J= 3.4 Hz, 1H), 6.94 (d,J= 7.2 Hz, 1H), 6.72 (d,J= 1.6 Hz, 1H), 4.66 (s, 2H), 4.38-4.28 (m, 1H), 4.05-3.83 (m, 4H), 3.50-3.47 (m, 1H), 3.24-3.08 (m, 2H), 2.57 (s, 3H), 2.06-1.89 (m, 3H), 1.81-1.73 (m,J= 12.6 Hz, 1H), 1.35 (d,J= 6.5 Hz, 3H). ESI-MS m / z 541.0 [M+H + ]

[0593]

[0594] Compound 41. (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(6-chloropyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride

[0595] Step 1) Preparation of 2-chloro-1-(6-chloropyridin-2-yl)ethan-1-one (86)

[0596] To a solution of 6-bromo-2-pyridinecarboxylic acid (700 mg, 4.44 mmol, 1.0 eq) in DCM (20 mL) at room temperature, oxalyl chloride (0.46 mL, 5.33 mmol, 1.2 eq) and DMF (5 drops) were added. The reaction mixture was stirred at room temperature for 2 h and then evaporated using a Rotavapor. Crude 6-chloropicolinoyl chloride (750 mg) was used in the next reaction without further purification.

[0597] To a solution of 6-chloropicolinoyl chloride (750 mg, 4.26 mmol, 1.0 eq) in 1,4-dioxane (15 mL) at 0°C was added 0.6 M TMS diazomethane (14.2 mL, 8.52 mmol, 2.0 eq). After stirring at room temperature for 16 h, 4 N HCl dissolved in 1,4-dioxane (6 mL) was added to the mixture, and the reaction mixture was stirred at room temperature for 2 h. The resulting mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography using Hep / EtOAc (8 / 2) as an eluent to give 86 (418 mg, 2.20 mmol, 50%, 2 steps) as a white solid.

[0598] 1 H NMR (500 MHz, CDCl3) δ 8.05 (d,J= 7.6 Hz, 1H), 7.88 (t,J= 7.8 Hz, 1H), 7.59 (d,J= 8.0 Hz, 1H), 5.10 (s, 2H). ESI-MS m / z 189.1 [M+H + ]

[0599] Step 2) Preparation of tert-butyl ((3S,4S)-8-(5-((8-chloro-2-(6-chloropyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)-6-methylpyrazin-2-yl)-3-methyl-2-oxa-8-azaspiro[4.5]decan-4-yl)carbamate (87)

[0600] To a solution of 8 (300 mg, 0.54 mmol, 1.0 eq) in ethanol (20 mL) were added 86 (123 mg, 0.65 mmol, 1.2 eq) and KI (18 mg, 0.11 mmol, 0.2 eq) at room temperature. The reaction mixture was stirred at 65°C for 16 h and concentrated under reduced pressure. The residue was then purified by silica gel column chromatography using DCM / MeOH (19 / 1) as the eluent to give 87 (80 mg, 0.12 mmol, 21%) as a white solid.

[0601] 1H NMR (400 MHz, MeOD) δ 8.44 (s, 1H), 8.29 (d,J= 7.2 Hz, 1H), 8.16 (d,J= 7.6 Hz, 1H), 7.90 (t,J= 7.8 Hz, 1H), 7.39 (d,J= 8.0 Hz, 1H), 6.54 (d,J= 7.2 Hz, 1H), 4.61 (s, 2H), 4.30-4.23 (m, 1H), 4.02-3.96 (m, 1H), 3.76 (d,J= 8.7 Hz, 1H), 3.69 (d,J= 8.7 Hz, 1H), 3.60-3.40 (m, 3H), 3.28 (s, 1H), 2.53 (s, 3H), 1.95-1.80 (m, 3H), 1.73-1.64 (m, 1H), 1.49 (d,J= 5.2 Hz, 9H), 1.17 (d,J= 6.3 Hz, 3H). ESI-MS m / z 686.1 [M+H + ]

[0602] Step 3) Preparation of (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(6-chloropyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 41)

[0603] To a solution of 87 (10 mg, 0.015 mmol, 1.0 eq) in DCM (5.0 mL) at 0°C was added 4N-HCl (0.04 mL, 0.016 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 1 h, and the solvent was evaporated in vacuo. The slurry was diluted with methanol (0.3 mL) and ethyl acetate (1.5 mL). After stirring for several minutes, the resulting solid was collected by filtration, washed with hexane, and dried in vacuo to obtain compound 41 (6.0 mg, 0.010 mmol, 66%) as a pale yellow solid.

[0604] 1H NMR (400 MHz, MeOD) δ 8.70 (s, 1H), 8.51 (d,J= 7.2 Hz, 1H), 8.13 (d,J= 7.6 Hz, 1H), 8.01 (t,J= 7.9 Hz, 1H), 7.56 (d,J= 8.0 Hz, 1H), 6.90 (d,J= 7.2 Hz, 1H), 4.66 (s, 2H), 4.38-4.29 (m, 1H), 4.06-3.82 (m, 4H), 3.51-3.46 (m, 1H), 3.23-3.06 (m, 2H), 2.57 (s, 3H), 1.99 (dd,J= 23.6, 12.0 Hz, 3H), 1.80-1.72 (m, 1H), 1.35 (d,J= 6.5 Hz, 3H). ESI-MS m / z 585.0 [M+H + ]

[0605]

[0606] Compound 42. (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(6-methoxypyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol hydrochloride

[0607] Step 1) Preparation of tert-butyl(S)-(1'-(5-bromo-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (88)

[0608] To a solution of 80 (5.42 g, 10.47 mmol, 1.0 eq) in DCM (85 mL) at -78°C was added dropwise 1 M DIBAL-H (52.5 mL, 52.5 mmol) in THF. After stirring at -78°C for 0.5 h, the bath was warmed to room temperature and stirred for another 0.5 h at room temperature. The reaction mixture was cooled to -78°C and added dropwise to a saturated aqueous Rochelle salt solution to quench the reaction. The resulting slurry was warmed to room temperature with vigorous stirring for 15 min, and then DCM and water were added. The mixture was extracted three times with DCM, and the combined organic layers were dried over anhydrous Na2SO4. The extracts were evaporated under reduced pressure. The resulting residue was purified by column chromatography (EtOAc / n-hexane = 15 / 100) to obtain 88 (2.50 g, 5.108 mmol, 49%) as a pale yellow solid.

[0609] Step 2) Preparation of tert-butyl(S)-(1'-(5-((2-amino-3-chloropyridin-4-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (89)

[0610] To a solution of 88 (2.40 g, 4.90 mmol, 1.0 eq) in dioxane (5 mL) at room temperature were added Pd(dba)3 (224 mg, 0.245 mmol, 0.05 eq), xantphos (284 mg, 0.490 mmol, 0.10 eq), DIEA (1.28 mL, 7.35 mmol, 1.5 eq), and 2-amino-3-chloro-pyridine-4-thio sodium salt (984 mg, 5.39 mmol, 1.1 eq). The reaction mixture was stirred at 95 °C for 16 h. The reaction was quenched with water and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using Hep / EtOAc (2 / 8) as the eluent to obtain 89 (1.60 g, 2.81 mmol, 57%) as a pale yellow solid.

[0611] 1 H NMR (500 MHz, MeOD) δ 8.30 (s, 1H), 7.63 (d,J= 5.6 Hz, 1H), 7.30-7.16 (m, 4H), 6.07 (d,J= 5.6 Hz, 1H), 4.94 (s, 1H), 4.69 (s, 2H), 3.87-3.75 (m, 2H), 3.31-3.26 (m, 1H), 3.14 (d,J= 15.7 Hz, 1H), 2.83 (d,J= 15.7 Hz, 1H), 2.01-1.93 (m, 1H), 1.89-1.81 (m, 1H), 1.80-1.68 (m, 1H), 1.55-1.46 (m, 10H), 1.37-1.33 (m, 1H). ESI-MS m / z 569.2 [M+H + ]

[0612] Step 3) Preparation of 2-bromo-1-(6-methoxypyridin-2-yl)ethan-1-one (90)

[0613] 1-(6-Methoxy-2-pyridinyl)ethanone (200 mg, 1.32 mmol, 1.0 eq) was dissolved in chloroform (5 mL) at room temperature, and 5 drops of HBr dissolved in acetic acid were added. A chloroform solution containing Br2 (0.08 mL, 1.58 mmol, 1.2 eq) was added dropwise through an addition funnel. The reaction mixture was stirred at room temperature for 16 h and then concentrated under reduced pressure. The residue was then purified by silica gel column chromatography using n-Hex / EtOAc (8 / 2) as an eluent to give 90 (172 mg, 0.75 mmol, 57%) as a white solid.

[0614] 1 H NMR (400 MHz, CDCl3) δ 7.79-7.70 (m, 2H), 7.01 (dd,J= 7.6, 1.5 Hz, 1H), 4.82 (s, 2H), 4.02 (s, 3H). ESI-MS m / z 229.1 [M+H + ]

[0615] Step 4) Preparation of tert-butyl(S)-(1'-(5-((8-chloro-2-(6-methoxypyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (91)

[0616] To a solution of 89 (60 mg, 0.11 mmol, 1.0 eq) in acetonitrile (5.0 mL) at room temperature were added 90 (49 mg, 0.22 mmol, 2.0 eq) and NaHCO3 (37 mg, 0.44 mmol, 4.0 eq). The reaction mixture was stirred at 80°C for 16 h and then cooled to room temperature. The reaction was quenched with water and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (19 / 1) as an eluent to give 91 (53 mg, 0.076 mmol, 73%) as a yellow solid.

[0617] 1 H NMR (400 MHz, MeOD) δ 8.44 (s, 1H), 8.32 (d,J= 7.2 Hz, 1H), 8.29 (s, 1H), 7.81-7.74 (m, 2H), 7.23 (s, 4H), 6.77 (dd,J= 7.7, 1.4 Hz, 1H), 6.70 (d,J= 7.2 Hz, 1H), 4.67 (s, 2H), 4.61 (s, 1H), 4.02 (s, 3H), 3.79-3.68 (m, 2H), 3.29-3.23 (m, 2H), 3.11 (s, 1H), 2.82 (d,J= 15.9 Hz, 1H), 2.07-1.94 (m, 2H), 1.89-1.80 (m, 1H), 1.76-1.71 (m, 1H), 1.50 (s, 89). ESI-MS m / z 699.1 [M+H + ]

[0618] Step 5) Preparation of (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((8-chloro-2-(6-methoxypyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol hydrochloride (Compound 42)

[0619] To a solution of 91 (50 mg, 0.071 mmol, 1.0 eq) in DCM (5.0 mL) at 0°C was added 4N-HCl (0.18 mL, 0.710 mmol, 10 eq) in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 h, and the solvent was evaporated in vacuo. The residue was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as the eluent to give compound 42 (24 mg, 0.038 mmol, 53%) as a pale yellow solid.

[0620] 1H NMR (400 MHz, MeOD) δ 8.73 (s, 1H), 8.59 (d,J= 7.2 Hz, 1H), 8.45 (s, 1H), 7.94-7.85 (m, 1H), 7.73 (d,J= 7.3 Hz, 1H), 7.54 (d,J= 7.4 Hz, 1H), 7.46-7.33 (m, 3H), 7.14 (d,J= 7.2 Hz, 1H), 6.97 (d,J= 8.3 Hz, 1H), 4.74 (s, 2H), 4.47 (s, 1H), 4.09 (s, 3H), 4.03 (d,J=14.4 Hz, 1H), 3.91 (d,J= 13.3 Hz, 1H), 3.46-3.35 (m, 2H), 3.21 (s, 2H), 2.10-2.02 (m, 1H), 1.97-1.89 (m, 1H), 1.86-1.78 (m, 1H), 1.71-1.63 (m, 1H). ESI-MS m / z 600.9 [M+H + ]

[0621]

[0622] Compound 43. tert-Butyl((3S,4S)-8(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(piperidin-4-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol dihydrochloride

[0623] Step 1) Preparation of tert-butyl(S)-4-(7-((5-(1-((tert-butoxycarbonyl)amino)-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)piperidine-1-carboxylate (92)

[0624] To a solution of 89 (350 mg, 0.62 mmol, 1.0 eq) in acetonitrile (20 mL) were added tert-Butyl 4-(2-bromoacetyl)piperidine-1-carboxylate (377 mg, 0.23 mmol, 2.0 eq) and NaHCO3 (221 mg, 2.48 mmol, 4.0 eq) at room temperature. The reaction mixture was stirred at 80 °C for 16 h. The reaction was quenched with water and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The crude mixture was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 92 (400 mg, 0.52 mmol, 84%) as a yellow solid.

[0625] 1 H NMR (400 MHz, MeOD) δ 8.27-8.15 (m, 2H), 7.68 (s, 1H), 7.22 (s, 4H), 6.65 (d, J= 7.2 Hz, 1H), 4.92 (s, 2H), 4.66 (s, 2H), 4.24-4.15 (m, 2H), 3.76-3.63 (m), 3.29-3.19 (m, 2H), 3.12 (d,J= 15.8 Hz, 1H), 3.00-2.91 (m, 2H), 2.81 (d,J= 15.7 Hz, 1H), 2.09 (d,J= 12.5 Hz, 2H), 2.00-1.92 (m, 1H), 1.87-1.80 (m, 1H), 1.77-1.58 (m, 4H), 1.55-1.46 (m, 18H). ESI-MS m / z 777.1 [M+H + ]

[0626] Step 2) Preparation of tert-butyl((3S,4S)-8(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(piperidin-4-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol dihydrochloride (compound 43)

[0627] To a solution of 92 (25 mg, 0.032 mmol, 1.0 eq) in DCM (5.0 mL) at 0°C was added 4N-HCl (0.08 mL, 0.32 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 h, and the solvent was evaporated in vacuo. The concentrate was purified using silica gel column chromatography. DCM / MeOH (9 / 1) was used as the eluent to obtain compound 43 (4.1 mg, 0.006 mmol, 20%) as a pale yellow solid.

[0628] 1 H NMR (500 MHz, MeOD) δ 8.54 (d,J= 7.2 Hz, 1H), 8.43 (s, 1H), 8.16 (d,J= 0.7 Hz, 1H), 7.55 (d,J= 7.5 Hz, 1H), 7.48-7.33 (m, 3H), 7.11 (d,J= 7.2 Hz, 1H), 4.73 (s, 2H), 4.47 (s, 1H), 4.08-3.98 (m, 1H), 3.94-3.86 (m, 1H), 3.63-3.55 (m, 2H), 3.43-3.35 (m, 2H), 3.30-3.24 (m, 2H), 3.21 (s, 2H), 2.47-2.35 (m, 2H), 2.12-2.01 (m, 3H), 1.97-1.89 (m, 1H), 1.85-1.76 (m, 1H), 1.72-1.65 (m, 1H). ESI-MS m / z 575.1 [M+H + ]

[0629]

[0630] Compound 44. (S)-1-(4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)piperidin-1-yl)ethan-1-one

[0631] Step 1) Preparation of (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(piperidin-4-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol (93)

[0632] To a solution of 92 (400 mg, 0.52 mmol, 1.0 eq) in DCM (5.0 mL) at 0°C was added 4N-HCl (1.30 mL, 5.20 mmol, 10 eq) dissolved in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 h and concentrated under reduced pressure. TEA (0.72 mL, 5.20 mmol, 10 eq) was added to the solution at 0°C, stirred at room temperature for 0.5 h, and then concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 93 (70 mg, 0.12 mmol, 24%) as a yellow oil.

[0633] 1 H NMR (500 MHz, MeOD) δ 8.54 (d,J= 7.2 Hz, 1H), 8.43 (s, 1H), 8.16 (d,J= 0.7 Hz, 1H), 7.55 (d,J= 7.5 Hz, 1H), 7.48-7.33 (m, 3H), 7.11 (d,J= 7.2 Hz, 1H), 4.73 (s, 2H), 4.47 (s, 1H), 4.08-3.98 (m, 1H), 3.94-3.86 (m, 1H), 3.63-3.55 (m, 2H), 3.43-3.35 (m, 2H), 3.30-3.24 (m, 2H), 3.21 (s, 2H), 2.47-2.35 (m, 2H), 2.12-2.01 (m, 3H), 1.97-1.89 (m, 1H), 1.85-1.76 (m, 1H), 1.72-1.65 (m, 1H). ESI-MS m / z 575.1 [M+H + ]

[0634] Step 2) Preparation of (S)-1-(4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)piperidin-1-yl)ethan-1-one (Compound 44)

[0635] To a solution of 93 (20 mg, 0.035 mmol, 1.0 eq) in DCM / MeOH (5.0 mL / 1.0 mL) at -20 °C were added TEA (3.9 uL, 0.028 mmol, 0.8 eq) and acetyl chloride (2.0 uL, 0.028 mmol, 0.8 eq). The reaction mixture was stirred at -20 °C for 0.5 h. The reaction was quenched with water and extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as the eluent to give compound 44 (2.3 mg, 0.004 mmol, 11%) as a yellow solid.

[0636] 1 H NMR (400 MHz, MeOD) δ 8.27-8.17 (m, 2H), 7.69 (s, 1H), 7.42-7.35 (m, 1H), 7.27-7.18 (m, 3H), 6.66 (d,J= 7.1 Hz, 1H), 4.70-4.58 (m, 3H), 4.07-3.97 (m, 2H), 3.81-3.71 (m, 2H), 3.30-3.18 (m, 3H), 3.17-3.03 (m, 2H), 2.88-2.76 (m, 2H), 2.22-2.09 (m, 5H), 2.00-1.87 (m, 2H), 1.81-1.66 (m, 2H), 1.65-1.60 (m,J1H), 1.51-1.45 (m, 1H). ESI-MS m / z 617.1 [M+H + ]

[0637]

[0638] Compound 45. Ethyl (S)-4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)piperidine-1-carboxylate

[0639] To a solution of 93 (20 mg, 0.035 mmol, 1.0 eq) in DCM / MeOH (5.0 mL / 1.0 mL) at -20 °C were added TEA (3.9 uL, 0.028 mmol, 0.8 eq) and ethyl chloroformate (2.7 uL, 0.028 mmol, 0.8 eq). The reaction mixture was stirred at -20 °C for 0.5 h. The reaction was quenched with water and extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as the eluent to give compound 45 (1.4 mg, 0.002 mmol, 6%) as a yellow solid.

[0640] 1 H NMR (400 MHz, MeOD) δ 8.27-8.18 (m, 2H), 7.69 (s, 1H), 7.45-7.37 (m, 1H), 7.30-7.22 (m, 3H), 6.67 (d,J= 7.1 Hz, 1H), 4.67 (s, 2H), 4.24 (d,J= 13.0 Hz, 2H), 4.19-4.11 (m, 2H), 3.81-3.71 (m, 2H), 3.29-3.19 (m, 2H), 3.19-3.11 (m, 1H), 3.06-2.94 (m, 3H), 2.92-2.83 (m, 1H), 2.13-1.89 (m, 5H), 1.72-1.61 (m, 3H), 1.56-1.49 (m, 1H), 1.29 (t,J= 7.1 Hz, 3H). ESI-MS m / z 647.9 [M+H + ]

[0641]

[0642] Compound 46. (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((8-chloro-2-(5-methoxypyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol hydrochloride

[0643] Step 1) Preparation of 2-bromo-1-(5-methoxypyridin-2-yl)ethan-1-one (95)

[0644] To a solution of 2-bromo-5-methoxypyridine (200 mg, 0.53 mmol, 1.0 eq) in 1,4-dioxane (5 mL) at room temperature were added Pd(PPh3)2Cl2 (19 mg, 0.027 mmol, 0.05 eq) and tributyl(1-ethoxyvinyl)stannane (0.20 mL, 0.58 mmol, 1.1 eq). The reaction mixture was stirred at 95 °C for 16 h. The reaction was quenched with water and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was briefly purified by silica gel column chromatography using Hep / EtOAc (9 / 1) as the eluent to give 94 (90 mg, 0.50 mmol, 47%) as a pale yellow solid. The unstable product was used in the next reaction without further purification.

[0645] To a solution of 94 (90 mg, 0.50 mmol, 1.0 eq) in THF (3.0 mL) and H2O (1.0 mL) was added NBS (93 mg, 0.53 mmol, 1.05 eq) at room temperature. The reaction mixture was stirred at room temperature for 1 h. The reaction was quenched with NaHCO3 and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using Hep / EtOAc (8 / 2) as the eluent to give 95 (34 mg, 0.15 mmol, 29%) as a pale yellow solid.

[0646] 1H NMR (400 MHz, CDCl3) δ 8.35 (d,J= 2.8 Hz, 1H), 8.13 (d,J= 8.7 Hz, 1H), 7.31 (dd,J= 8.7, 2.9 Hz, 1H), 4.83 (s, 2H), 3.97 (s, 3H). ESI-MS m / z 228.9 [M+H + ]

[0647] Step 2) Preparation of tert-butyl(S)-(1'-(5-((8-chloro-2-(5-methoxypyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (95)

[0648] To a solution of 89 (45 mg, 0.08 mmol, 1.0 eq) in acetonitrile (5.0 mL) at room temperature were added 95 (27 mg, 0.23 mmol, 1.5 eq) and NaHCO3 (77 mg, 0.92 mmol, 4.0 eq). The reaction mixture was stirred at 80 °C for 16 h. The reaction was quenched with water (10 mL) and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 96 (23 mg, 0.033 mmol, 42%) as a yellow solid.

[0649] 1H NMR (400 MHz, MeOD) δ 8.35-8.25 (m, 4H), 8.18 (d,J= 8.7 Hz, 1H), 7.53 (dd,J= 8.8, 3.0 Hz, 1H), 7.22 (s, 4H), 7.05 (d,J= 10.1 Hz, 1H), 6.68 (d,J= 7.2 Hz, 1H), 4.96-4.92 (m, 1H), 4.67 (s, 2H), 4.61 (s, 1H), 3.95 (s, 3H), 3.80-3.66 (m, 2H), 3.30-3.22 (m, 2H), 3.13 (d,J= 15.8 Hz, 1H), 2.81 (d,J= 15.8 Hz, 1H), 2.02-1.92 (m, 1H), 1.89-1.81 (m, 1H), 1.77-1.70 (m, 1H), 1.50 (s, 10H). ESI-MS m / z 699.1 [M+H + ]

[0650] Step 3) Preparation of (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((8-chloro-2-(5-methoxypyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol hydrochloride (compound 46)

[0651] To a solution of 96 (23 mg, 0.033 mmol, 1.0 eq) in DCM (5.0 mL) at 0°C was added 4N-HCl (0.08 mL, 0.33 mmol, 10 eq) in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 h, and the solvent was evaporated in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (19 / 1) as the eluent to give compound 46 (1.9 mg, 0.003 mmol, 10%) as a pale yellow solid.

[0652] 1H NMR (400 MHz, MeOD) δ 8.66 (s, 1H), 8.54-8.46 (m, 2H), 8.41 (s, 1H), 8.21 (d,J= 8.9 Hz, 1H), 7.81 (dd,J= 8.9, 2.8 Hz, 1H), 7.54 (d,J= 7.6 Hz, 1H), 7.44-7.36 (m, 3H), 7.01 (d,J= 7.2 Hz, 1H), 4.73 (s, 2H), 4.46 (s, 1H), 4.07-3.96 (m, 4H), 3.91-3.84 (m, 1H), 3.42-3.35 (m, 2H), 3.21 (s, 2H), 2.05 (t,J= 10.1 Hz, 1H), 1.97-1.89 (m, 1H), 1.85-1.78 (m, 1H), 1.72-1.64 (m, 1H). ESI-MS m / z 599.1 [M+H + ]

[0653]

[0654] Compound 47. (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(2-methoxypyrimidin-4-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol hydrochloride

[0655] Step 1) Preparation of 2-bromo-1-(2-methoxypyrimidin-4-yl)ethan-1-one (97)

[0656] To a solution of 4-bromo-2-methoxypyrimidine (100 mg, 0.53 mmol, 1.0 eq) in 1,4-dioxane (5.0 mL) at room temperature were added Pd(PPh3)2Cl2 (19 mg, 0.027 mmol, 0.05 eq) and tributyl(1-ethoxyvinyl)stannane (0.20 mL, 0.58 mmol, 1.1 eq). The reaction mixture was stirred at 95 °C for 3 h. The reaction was quenched with water (10 mL) and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was briefly purified by silica gel column chromatography using Hep / EtOAc (9 / 1) as the eluent to give 4-(1-ethoxyvinyl)-2-methoxypyrimidine (84 mg, 0.47 mmol, 88%) as a pale yellow solid. The unstable product was used in the next reaction without further purification.

[0657] To a solution of 4-(1-ethoxyvinyl)-2-methoxypyrimidine (84 mg, 0.47 mmol, 1.0 eq) in THF (3.0 mL) and H2O (1.0 mL) was added NBS (87 mg, 0.49 mmol, 1.05 eq) at room temperature. The reaction mixture was stirred at room temperature for 1 h. The reaction was quenched with NaHCO3 and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using Hep / EtOAc (8 / 2) as an eluent to give 97 (80 mg, 0.35 mmol, 74%) as a pale yellow solid.

[0658] 1 H NMR (500 MHz, CDCl3) δ 8.88-8.70 (m, 1H), 7.66-7.49 (m, 1H), 4.76 (s, 2H), 4.12 (s, 3H). ESI-MS m / z 229.8 [M+H + ]

[0659] Step 2) Preparation of tert-butyl(S)-(1'-(5-((8-chloro-2-(2-methoxypyrimidin-4-yl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (98)

[0660] To a solution of 89 (35 mg, 0.061 mmol, 1.0 eq) in acetonitrile (5.0 mL) at room temperature were added 97 (22 mg, 0.122 mmol, 2.0 eq) and NaHCO3 (20 mg, 0.244 mmol, 4.0 eq). The reaction mixture was stirred at 80 °C for 16 h. The reaction was quenched with water (10 mL) and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 98 (26 mg, 0.037 mmol, 60%) as a yellow solid.

[0661] 1 H NMR (400 MHz, MeOD) δ 8.63 (d,J= 5.1 Hz, 1H), 8.57 (s, 1H), 8.29 (d,J= 7.1 Hz, 2H), 7.84 (d,J= 5.1 Hz, 1H), 7.22 (s, 4H), 6.67 (d,J= 7.2 Hz, 1H), 4.93 (s, 1H), 4.67 (s, 2H), 4.12-4.06 (m, 3H), 3.81-3.66 (m, 2H), 3.31-3.21 (m, 2H), 3.12 (d,J= 15.9 Hz, 1H), 2.81 (d,J= 15.7 Hz, 1H), 2.01-1.91 (m1H), 1.90-1.80 (m, 1H), 1.76-1.69 (m, 1H), 1.59-1.44 (m, 10H). ESI-MS m / z 701.1 [M+H + ]

[0662] Step 3) Preparation of (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((8-chloro-2-(2-methoxypyrimidin-4-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol hydrochloride (Compound 47)

[0663] To a solution of 98 (22 mg, 0.031 mmol, 1.0 eq) in DCM (5.0 mL) at 0°C was added 4N-HCl (0.08 mL, 0.31 mmol, 10 eq) in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 h, and the solvent was evaporated in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (19 / 1) as the eluent to give compound 47 (1.8 mg, 0.003 mmol, 10%) as a pale yellow solid.

[0664] 1 H NMR (400 MHz, MeOD) δ 8.95 (s, 1H), 8.81 (d,J= 5.3 Hz, 1H), 8.61-8.56 (m, 1H), 8.46-8.41 (m, 1H), 7.87 (d,J= 5.3 Hz, 1H), 7.55 (d,J= 7.5 Hz, 1H), 7.44-7.35 (m, 3H), 7.09 (d,J= 7.3 Hz, 1H), 4.73 (d,J= 6.8 Hz, 2H), 4.47 (s, 1H), 4.24-4.15 (m, 3H), 4.05-3.97 (m, 1H), 3.96-3.88 (m, 1H), 3.37 (s, 3H), 3.21 (s, 2H), 2.11-2.02 (m, 1H), 1.99-1.89 (m, 1H), 1.87-1.79 (m, 1H), 1.71-1.64 (m, 1H). ESI-MS m / z 600.8 [M+H + ]

[0665]

[0666] Compound 48. (S)-4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-N-(2-methoxyethyl)benzamide

[0667] Step 1) Preparation of ethyl (S)-7-((5-(1-((tert-butoxycarbonyl)amino)-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)benzoate (99)

[0668] To a solution of 89 (500 mg, 0.88 mmol, 1.0 eq) in acetonitrile (20 mL) at room temperature were added 53 (477 mg, 1.76 mmol, 2.0 eq) and NaHCO3 (260 mg, 3.25 mmol, 4.0 eq). The reaction mixture was stirred at 85 °C for 16 h and then cooled to room temperature. The solvent was removed in vacuo, and the residue was diluted with water (20 mL). The mixture was extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 99 (500 mg, 0.67 mmol, 77%) as a yellow solid.

[0669] 1H NMR (500 MHz, MeOD) δ 8.34 (s, 1H), 8.31-8.24 (m, 2H), 8.07 (d,J= 8.5 Hz, 2H), 7.94-7.88 (m, 2H), 7.27-7.16 (m, 4H), 7.06 (d,J= 9.9 Hz, 1H), 6.69 (d,J= 7.2 Hz, 1H), 4.93 (d,J= 10.0 Hz, 1H), 3.79-3.66 (m, 3H), 3.49-3.42 (m, 2H), 3.30-3.20 (m, 3H), 3.13 (d,J= 15.7 Hz, 1H), 2.98-2.92 (m, 3H), 2.81 (d, 1H), 2.00-1.93 (m, 1H), 1.88-1.81 (m, 1H), 1.76-1.68 (m, 1H), 1.54-1.48 (m, 9H). ESI-MS m / z 741.2 [M+H + ]

[0670] Step 2) Preparation of (S)-7-((5-(1-((tert-butoxycarbonyl)amino)-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridine-2-carboxylic acid (100)

[0671] LiOH-H2O (10 mg, 0.45 mmol, 3.0 eq) was added to a solution of 99 (100 mg, 0.15 mmol, 1.0 eq) dissolved in a mixture of THF (4.0 mL), MeOH (1.0 mL), and H2O (1.0 mL) at room temperature. The resulting mixture was stirred at room temperature for 12 h and concentrated under reduced pressure. The residue was diluted with H2O (3 mL), and the pH was adjusted to 4 by adding 1 M HCl to the H2O. The precipitate was collected by filtration with n-heptane, and the solid was dried in vacuo. The crude product 100 was used in the next reaction.

[0672] ESI-MS m / z 712.1 [M+H + ]

[0673] Step 3) Preparation of tert-butyl (S)-(1'-(5-((8-chloro-2-((2-methoxyethyl) carbamoyl) imidazo[1,2-a] pyridin-7-yl) thio)-3-(hydroxymethyl) pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl) carbamate (101)

[0674] To a solution of 100 (30 mg, 0.042 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added 2-methoxyethylamine (4.7 μL, 0.055 mmol, 1.3 eq), HATU (24 mg, 0.063 mmol, 1.5 eq), and DIEA (14.7 μL, 0.084 mmol, 2.0 eq). The resulting mixture was stirred at room temperature for 12 h. The reaction was quenched with water and brine, then extracted with EtOAc (2 X 15 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 101 (14 mg, 0.018 mmol, 44%) as a yellow solid.

[0675] 1 H NMR (400 MHz, MeOD) δ 8.35-8.25 (m, 3H), 8.07 (d,J= 8.5 Hz, 2H), 7.93 (d,J= 8.5 Hz, 2H), 7.22 (s, 4H), 6.69 (d,J= 7.2 Hz, 1H), 4.96-4.91 (m, 1H), 4.67 (s, 2H), 3.80-3.67 (m, 2H), 3.61 (s, 4H), 3.42 (s, 3H), 3.30-3.22 (m, 2H), 3.13 (d, 1H), 2.81 (d,J= 16.0 Hz, 1H), 2.10-1.92 (m, 2H), 1.89-1.80 (m, 1H), 1.77-1.69 (m, 1H), 1.50 (s, 10H). ESI-MS m / z 769.1 [M+H + ]

[0676] Step 4) Preparation of (S)-4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-N-(2-methoxyethyl)benzamide (Compound 48)

[0677] To a solution of 101 (14 mg, 0.018 mmol, 1.0 eq) in DCM (5.0 mL) at 0°C was added 4N-HCl (0.05 mL, 0.18 mmol, 10 eq) in 1,4-dioxane. The resulting mixture was stirred at room temperature for 2 h, and the solvent was evaporated in vacuo. The residue was quenched with NaHCO3 and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give compound 48 (2.8 mg, 0.004 mmol, 23%) as a pale yellow solid.

[0678] 1 H NMR (500 MHz, MeOD) δ 8.33 (s, 1H), 8.29 (d,J= 5.9 Hz, 2H), 8.08 (d,J= 8.5 Hz, 2H), 7.93 (d,J= 8.5 Hz, 2H), 7.42-7.37 (m, 1H), 7.27-7.20 (m, 3H), 6.70 (d,J= 7.2 Hz, 1H), 4.69 (s, 2H), 4.03 (s, 1H), 3.82-3.73 (m, 2H), 3.61 (s, 4H), 3.44-3.40 (m, 3H), 3.29-3.21 (m, 2H), 3.15 (d,J= 15.7 Hz, 1H), 2.84 (d,J= 15.7 Hz, 1H), 2.08-1.90 (m, 3H), 1.67 - 1.61 (d,J= 13.1 Hz, 1H), 1.53 - 1.47 (d,J= 13.1 Hz, 1H). ESI-MS m / z 700.1 [M+H + ]

[0679]

[0680] Compound 49. (S)-4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-N-methylbenzamide

[0681] Step 1) Preparation of tert-butyl(S)-(1'-(5-((8-chloro-2-(4-(methylcarbamoyl)phenyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (102)

[0682] To a solution of 100 (30 mg, 0.042 mmol, 1.0 eq) in DMF (1.0 mL) at room temperature were added methylamine HCl salt (3.7 mg, 0.055 mmol, 1.3 eq), HATU (24 mg, 0.063 mmol, 1.5 eq), and DIEA (22 μL, 0.126 mmol, 3.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 102 (25 mg, 0.034 mmol, 70%) as a yellow solid.

[0683] 1H NMR (500 MHz, MeOD) δ 8.33 (s, 1H), 8.31-8.24 (m, 2H), 8.07 (d,J= 8.5 Hz, 2H), 7.94-7.88 (m, 2H), 7.27-7.16 (m, 4H), 7.06 (d,J= 9.9 Hz, 1H), 6.69 (d,J= 7.2 Hz, 1H), 4.93 (d,J= 10.0 Hz, 1H), 4.67 (s, 2H), 3.79-3.66 (m, 3H), 3.30-3.20 (m, 3H), 3.13 (d,J= 15.7 Hz, 1H), 2.98-2.92 (m, 3H), 2.81 (d, 1H), 2.00-1.93 (m, 1H), 1.88-1.81 (m, 1H), 1.76-1.68 (m, 1H), 1.54-1.48 (m, 9H). ESI-MS m / z 726.2 [M+H + ]

[0684] Step 2) Preparation of (S)-4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-N-methylbenzamide (Compound 49)

[0685] To a solution of 102 (20 mg, 0.028 mmol, 1.0 eq) in DCM (5.0 mL) at 0°C was added 4N-HCl (0.07 mL, 0.28 mmol, 10 eq) in 1,4-dioxane. The resulting mixture was stirred at room temperature for 2 h, and the solvent was evaporated in vacuo. The mixture was quenched with NaHCO3 and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give compound 49 (5.4 mg, 0.009 mmol, 31%) as a pale yellow solid.

[0686] 1H NMR (400 MHz, MeOD) δ 8.32 (s, 1H), 8.28 (t,J= 3.6 Hz, 2H), 8.06 (d,J= 8.4 Hz, 2H), 7.91 (d,J= 8.4 Hz, 2H), 7.44-7.37 (m, 1H), 7.30-7.19 (m, 3H), 6.69 (d,J= 7.2 Hz, 1H), 4.72-4.63 (m, 2H), 4.05 (s, 1H), 3.84-3.72 (m, 2H), 3.30-3.20 (m, 2H), 3.14 (d,J= 15.8 Hz, 1H), 2.96 (s, 3H), 2.85 (d,J= 15.8 Hz, 1H), 2.06-1.89 (m, 2H), 1.63 (d,J= 12.5 Hz, 1H), 1.51 (d,J= 13.2 Hz, 1H). ESI-MS m / z 626.2 [M+H + ]

[0687]

[0688] Compound 50. (S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(4-aminophenyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol

[0689] Step 1) Preparation of tert-butyl (S)-(1'-(5-((8-chloro-2-(4-nitrophenyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (103)

[0690] 89 (500 mg, 0.88 mmol, 1.0 eq) was dissolved in acetonitrile (20 mL) at room temperature, 2-bromo-1-(4-nitrophenyl)ethan-1-one (430 mg, 1.76 mmol, 2.0 eq) and NaHCO3 (260 mg, 3.25 mmol, 4.0 eq) were added. The reaction mixture was stirred at 85 °C for 16 h and then cooled to room temperature. The solvent was removed in vacuo, and the residue was diluted with water (20 mL). The mixture was extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 103 (447 mg, 0.63 mmol, 71%) as a yellow solid.

[0691] 1 H NMR (400 MHz, CDCl3) δ 8.33-8.26 (m, 2H), 8.25 (s, 1H), 8.17-8.10 (m, 2H), 7.98 (s, 1H), 7.96 (d,J= 7.1 Hz, 1H), 7.31-7.27 (m, 1H), 7.23 (dq,J= 7.9, 2.6 Hz, 3H), 6.62 (d,J= 7.1 Hz, 1H), 5.00 (d,J= 9.8 Hz, 1H), 4.66 (d,J= 8.7 Hz, 3H), 3.82 (s, 1H), 3.56 (d,J=13.2 Hz, 2H), 3.17 (td,J= 11.2, 9.6, 4.6 Hz, 2H), 3.03 (d,J= 15.8 Hz, 1H), 2.80 (d,J= 15.8 Hz, 1H), 2.04-1.95 (m, 1H), 1.87-1.78 (m, 1H), 1.73 (d,J= 13.5 Hz, 1H), 1.64 (s, 1H), 1.48 (s, 9H). ESI-MS m / z 715 [M+H + ]

[0692] Step 2) Preparation of tert-butyl(S)-(1'-(5-((2-(4-aminophenyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (104)

[0693] To a solution of 103 (447 mg, 0.63 mmol, 1.0 eq) in H2O / EtOH (1 / 4) were added Fe (175 mg, 3.13 mmol, 5.0 eq) and NH4Cl (167 mg, 3.13 mmol, 5.0 eq). The reaction mixture was refluxed for 2 h and filtered. The prepared mixture was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as the eluent to obtain 104 (273 mg, 0.40 mmol, 64%) as a yellow oil.

[0694] 1 H NMR (400 MHz, CDCl3) δ 8.20 (s, 1H), 7.94 (d, J= 7.0 Hz, 1H), 7.80 (d, 2H), 7.76 (s, 1H), 7.32 - 7.29 (m, 1H), 7.27-7.20 (m, 3H), 6.80-6.73 (m, 2H), 6.64 (d,J= 7.0 Hz, 1H), 5.02 (d,J= 9.8 Hz, 1H), 4.68 (d,J= 10.1 Hz, 3H), 3.77 (s, 2H), 3.53 (d,J= 12.9 Hz, 2H), 3.22-3.09 (m, 2H), 3.05 (d,J= 15.8 Hz, 1H), 2.81 (d,J= 15.9 Hz, 1H), 2.00 (d,J= 3.9 Hz, 1H), 1.89-1.79 (m, 1H), 1.75 (d,J= 13.7 Hz, 1H), 1.66 (s, 2H), 1.51 (s, 9H). ESI-MS m / z 685 [M+H + ]

[0695] Step 3) Preparation of (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(4-aminophenyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol (Compound 50)

[0696] To a solution of 104 (8 mg, 0.017 mmol, 1.0 eq) in DCM (5.0 mL) at 0°C was added 4N-HCl (0.09 mL, 0.34 mmol, 20 eq) in 1,4-dioxane. The resulting mixture was stirred at room temperature for 2 h, and the solvent was evaporated in vacuo. The mixture was quenched with NaHCO3 and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give compound 50 (2.0 mg, 0.003 mmol, 29%) as a yellow solid.

[0697] 1 H NMR (400 MHz, MeOD) δ 8.23-8.18 (m, 2H), 8.01 (s, 1H), 7.84 (s, 1H), 7.71-7.67 (m, 2H), 7.41 (d, J= 6.5 Hz, 1H), 7.28-7.22 (m, 3H), 6.81-6.75 (m, 2H), 6.68 (d,J= 7.2 Hz, 1H), 4.66 (s, 3H), 4.12 (s, 1H), 3.72 (t,J= 14.5 Hz, 2H), 3.22 (t,J= 12.2 Hz, 3H), 3.12 (d,J=16.0 Hz, 2H), 2.91 (d,J= 16.0 Hz, 2H), 2.03 (d,J= 9.1 Hz, 1H), 1.93 (t,J= 12.1 Hz, 2H), 1.59 (dd,J= 23.2, 13.5 Hz, 3H). ESI-MS m / z 584 [M+H + ]

[0698]

[0699] Compound 51. (S)-N-(4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenyl)acetamide

[0700] Step 1) Preparation of tert-Butyl (S)-(1'-(5-((2-(4-acetamidophenyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (105)

[0701] To a solution of 102 (32 mg, 0.073 mmol, 1.0 eq) in DCM (2.0 mL) at 0°C was added acetic anhydride (22 μL, 0.365 mmol, 5.0 eq). The reaction mixture was stirred at room temperature for 2 h, and the solvent was evaporated in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as the eluent to give 105 (32 mg, 0.044 mmol, 61%) as a yellow solid.

[0702] 1H NMR (400 MHz, CDCl3) δ 8.22 (s, 1H), 7.92 (d,J= 7.1 Hz, 1H), 7.88 (d,J= 8.3 Hz, 2H), 7.80 (s, 1H), 7.69 (s, 1H), 7.58 (d,J= 8.3 Hz, 2H), 7.30 (d,J= 5.2 Hz, 1H), 7.24 (dq,J= 7.8, 3.1, 2.5 Hz, 3H), 6.60 (d,J= 7.1 Hz, 1H), 5.02 (d,J= 9.8 Hz, 1H), 4.72 (d,J= 9.8 Hz, 1H), 4.66 (s, 2H), 3.61 - 3.48 (m, 2H), 3.23 - 3.07 (m, 2H), 3.04 (d,J= 15.8 Hz, 1H), 2.80 (d,J= 15.7 Hz, 1H), 2.19 (s, 3H), 2.10 - 1.97 (m, 2H), 1.88 - 1.80 (m, 1H), 1.74 (d,J= 13.5 Hz, 1H), 1.51 (s, 9H). ESI-MS m / z 727 [M+H + ]

[0703] Step 2) Preparation of (S)-N-(4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenyl)acetamide (Compound 51)

[0704] To a solution of 105 (8.0 mg, 0.011 mmol, 1.0 eq) in DCM (3.0 mL) at 0 °C was added 4N-HCl (0.03 mL, 0.11 mmol, 10 eq) in 1,4-dioxane. The resulting mixture was stirred at room temperature for 2 h, and the solvent was evaporated in vacuo. The residue was quenched with saturated aqueous NaHCO3 solution (2.0 mL), diluted with water (10 mL), and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give compound 51 (1.0 mg, 0.002 mmol, 15%) as a pale yellow solid.

[0705] 1 H NMR (400 MHz, MeOD) δ 8.28 (d,J= 7.1 Hz, 1H), 8.22 (d,J= 21.4 Hz, 2H), 7.94-7.87 (m, 2H), 7.66 (d,J= 8.6 Hz, 2H), 7.50 (d,J= 7.5 Hz, 1H), 7.43-7.30 (m, 3H), 6.74 (d,J= 7.2 Hz, 1H), 4.66 (s, 2H), 4.57 (s, 2H), 4.40 (s, 1H), 3.83 (d,J= 12.6 Hz, 1H), 3.73 (d,J= 13.5 Hz, 1H), 2.16 (d,J= 1.6 Hz, 3H), 2.07-1.96 (m, 3H), 1.89 (t,J= 11.4 Hz, 1H), 1.75 (d,J= 13.2 Hz, 1H), 1.64 (d,J= 14.4 Hz, 2H). ESI-MS m / z 626 [M+H + ]

[0706]

[0707] Compound 52. (S)-4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-2-fluoro-N-methylbenzamide hydrochloride

[0708] Step 1) Preparation of 4-bromo-2-fluorobenzoic acid (106)

[0709] To a solution of methyl 4-bromo-2-fluorobenzoate (150 mg, 0.64 mmol, 1.0 eq) in THF (4.0 mL) and MeOH (1.0 mL) at room temperature was added LiOH (46 mg, 1.92 mmol, 3.0 eq) dissolved in H2O (1.0 mL). The reaction mixture was stirred at room temperature for 1 h. TLC analysis showed complete disappearance of the starting material. The reaction mixture was concentrated to remove THF and methanol. The aqueous solution containing the product was diluted with water (5.0 mL), pH adjusted to 2 with 1 N-HCl (aq), and the mixture was extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo to give 106 (120 mg, 0.55 mmol, 86%) as a colorless oil.

[0710] ESI-MS m / z 218.1 [M+H + ]

[0711] Step 2) Preparation of 4-bromo-2-fluoro-N-methylbenzamide (107)

[0712] 106 (120 mg, 0.55 mmol, 1.0 eq) was dissolved in dichloromethane (5.0 mL), and the stirred suspension was added oxalyl chloride (0.24 mL, 2.75 mmol, 5.0 eq) and DMF (1 drop) in an ice bath. The reaction mixture was cooled to room temperature, stirred at room temperature for 2 h, and then evaporated in vacuo.

[0713] To a solution of the acid chloride in DCM (5.0 mL) in an ice bath were added triethylamine (0.38 mL, 2.75 mmol, 5.0 eq) and methylamine HCl salt (45 mg, 0.66 mmol, 1.2 eq). The reaction mixture was stirred at room temperature for 2 h, treated with H2O (5.0 mL), and extracted twice with DCM (10 mL). The organic layer was washed with 1 M HCl in H2O, dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using Hep / EtOAc (1 / 1) as the eluent to give 107 (64 mg, 0.28 mmol, 50%) as a white solid.

[0714] 1 H NMR (400 MHz, CDCl3) δ 8.02 (t,J= 8.5 Hz, 1H), 7.44 (dd,J= 8.4, 1.8 Hz, 1H), 7.34 (dd,J= 11.4, 1.8 Hz, 1H), 6.68 (s, 1H), 3.05 (dd,J= 4.8, 0.8 Hz, 3H). ESI-MS m / z 231.1 [M+H + ]

[0715] Step 3) Preparation of 4-(1-ethoxyvinyl)-2-fluoro-N-methylbenzamide (108)

[0716] To a solution of 107 (60 mg, 0.26 mmol, 1.0 eq) in 1,4-dioxane (5.0 mL) at room temperature were added Pd(PPh3)2Cl2 (9.1 mg, 0.013 mmol, 0.05 eq) and tributyl(1-ethoxyvinyl)stannane (0.10 mL, 0.29 mmol, 1.1 eq). The reaction mixture was stirred at 95 °C for 3 h and then cooled to room temperature. The reaction mixture was quenched with water (5.0 mL) and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using Hep / EtOAc (1 / 1) as the eluent to give 108 (48 mg, 0.22 mmol, 83%) as a pale yellow oil.

[0717] 1 H NMR (500 MHz, CDCl3) δ 8.24 (t,J= 7.9 Hz, 1H), 7.83 (dd,J= 8.1, 1.6 Hz, 1H), 7.72 (dd,J= 12.5, 1.5 Hz, 1H), 6.80 (s, 1H), 3.75 (q,J= 7.0 Hz, 2H), 3.08 (dd,J= 4.8, 0.9 Hz, 3H), 2.65 (s, 3H), 1.27 (t,J= 7.0 Hz, 3H).

[0718] Step 4) Preparation of 4-(2-bromoacetyl)-2-fluoro-N-methylbenzamide (109)

[0719] To a solution of 108 (45 mg, 0.20 mmol, 1.0 eq) in THF (3.0 mL) and H2O (1.0 mL) was added NBS (37 mg, 0.21 mmol, 1.05 eq) at room temperature. The reaction mixture was stirred at room temperature for 1 h. The reaction was quenched with saturated aqueous NaHCO3 solution (5.0 mL) and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using Hep / EtOAc (8 / 2) as the eluent to give 109 (37 mg, 0.13 mmol, 67%) as a pale yellow solid.

[0720] 1 H NMR (500 MHz, CDCl3) δ 8.27 (t,J= 7.9 Hz, 1H), 7.87 (dd,J= 8.1, 1.2 Hz, 1H), 7.77 (d,J= 12.2 Hz, 1H), 6.79 (s, 1H), 4.45 (s, 2H), 3.09 (d,J= 4.8 Hz, 3H). ESI-MS m / z 275.1 [M+H + ]

[0721] Step 5) Preparation of tert-butyl(S)-(1'-(5-((8-chloro-2-(3-fluoro-4-(methylcarbamoyl)phenyl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (110)

[0722] To a solution of 89 (30 mg, 0.053 mmol, 1.0 eq) in acetonitrile (5.0 mL) at room temperature were added 109 (22 mg, 0.080 mmol, 1.5 eq) and NaHCO3 (18 mg, 0.212 mmol, 4.0 eq). The reaction mixture was stirred at 80 °C for 16 h. The reaction was quenched with water and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 110 (18 mg, 0.0 24 mmol, 46%) as a yellow solid.

[0723] 1 H NMR (400 MHz, MeOD) δ 8.35 (s, 1H), 8.30-8.24 (m, 2H), 7.89-7.81 (m, 3H), 7.26-7.18 (m, 4H), 6.68 (d,J= 7.2 Hz, 1H), 3.79-3.65 (m, 2H), 3.32-3.21 (m, 2H), 3.13 (d,J= 15.9 Hz, 1H), 2.98 (s, 3H), 2.81 (d,J= 15.7 Hz, 1H), 2.08-1.92 (m, 3H), 1.90-1.79 (m, 1H), 1.78-1.70 (m, 1H), 1.55-1.43 (m, 10H). ESI-MS m / z 743.1 [M+H + ]

[0724] Step 6) Preparation of (S)-4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-2-fluoro-N-methylbenzamide hydrochloride (Compound 52)

[0725] At 0°C, 4N-HCl (0.06 mL, 0.24 mmol, 10 eq) dissolved in 1,4-dioxane was added to a solution of 110 (18 mg, 0.024 mmol, 1.0 eq) in DCM (5.0 mL). The resulting mixture was stirred at room temperature for 3 h, and the solvent was evaporated in vacuo. The solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as the eluent to give compound 52 (5.7 mg, 0.008 mmol, 35%) as a pale yellow solid.

[0726] 1 H NMR (400 MHz, MeOD) δ 8.60 (s, 1H), 8.52 (d,J= 7.2 Hz, 1H), 8.41 (s, 1H), 7.98-7.84 (m, 3H), 7.54 (d,J= 7.4 Hz, 1H), 7.47-7.35 (m, 3H), 7.05 (d,J= 7.3 Hz, 1H), 5.00-4.92 (m, 1H), 4.73 (s, 2H), 4.46 (s, 1H), 3.99 (d,J= 14.5 Hz, 1H), 3.88 (d,J= 13.8 Hz, 1H), 3.43-3.35 (m, 2H), 3.21 (s, 2H), 2.98 (s, 3H), 2.12-2.01 (m, 1H), 1.97-1.90 (m, 1H), 1.81 (d,J= 12.9 Hz, 1H), 1.68 (d,J= 14.2 Hz, 1H). ESI-MS m / z 643.4 [M+H + ]

[0727]

[0728] Compound 53. (S)-6-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-2-chloro-N-methylnicotinamide

[0729] Step 1) Preparation of 6-bromo-2-chloronicotinic acid (111)

[0730] To a solution of ethyl 6-bromo-2-chloronicotinate (1.00 g, 3.78 mmol, 1.0 eq) in THF (12 mL) and MeOH (4.0 mL) was added a solution of LiOH (271 mg, 11.34 mmol, 3.0 eq) in H2O (4.0 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 h. TLC analysis showed the complete disappearance of the starting material. The reaction mixture was concentrated to remove THF and methanol. The aqueous solution containing the product was adjusted to pH 2 with 1 N-HCl (aq), and the mixture was extracted with DCM (2 X 25 mL). The combined organic layers were dried over Na2SO4 and the solvent was removed in vacuo to obtain 111 (790 mg, 3.34 mmol, 88%) as a colorless oil.

[0731] 1 H NMR (400 MHz, CDCl3) δ 8.21 (d,J= 8.1 Hz, 1H), 7.59 (d,J= 8.1 Hz, 1H). ESI-MS m / z 235.1 [M+H + ]

[0732] Step 2) Preparation of 6-bromo-2-chloro-N-methylnicotinamide (112)

[0733] To a solution of 111 (790 mg, 3.34 mmol, 1.0 eq) in DMF (5.0 mL) at room temperature were added methylamine HCl salt (293 mg, 4.32 mmol, 1.3 eq), HATU (1.91 g, 5.01 mmol, 1.5 eq), and DIEA (1.75 mL, 10.02 mmol, 3.0 eq). The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water and brine, then extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using Hep / EtOAc (1 / 1) as the eluent to give 112 (218 mg, 0.92 mmol, 26%) as a yellow solid.

[0734] 1 H NMR (400 MHz, CDCl3) δ 7.94 (d,J= 8.0 Hz, 1H), 7.47 (d,J= 8.0 Hz, 1H), 6.48 (s, 1H), 2.98 (d,J= 4.9 Hz, 3H). ESI-MS m / z 248.1 [M+H + ]

[0735] Step 3) Preparation of 2-chloro-6-(1-ethoxyvinyl)-N-methylnicotinamide (113)

[0736] To a solution of 112 (218 mg, 0.92 mmol, 1.0 eq) in dioxane (5 mL) at room temperature were added Pd(PPh3)2Cl2 (65 mg, 0.092 mmol, 0.1 eq), CuI (26 mg, 0.138 mmol, 0.15 eq), and tributyl(1-ethoxyvinyl)stannane (0.34 mL, 1.01 mmol, 1.1 eq). The reaction mixture was stirred at 80 °C for 3 h. The reaction was quenched with water and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using Hep / EtOAc (1 / 1) as the eluent to obtain 113 (150 mg, 0.62 mmol, 71%) as a pale yellow oil.

[0737] 1 H NMR (400 MHz, CDCl3) δ 8.22 (d,J= 7.8 Hz, 1H), 8.04 (d,J= 7.8 Hz, 1H), 3.12-3.03 (m, 4H), 2.73 (s, 3H), 1.27-1.17 (m, 3H).

[0738] Step 4) Preparation of 6-(2-bromoacetyl)-2-chloro-N-methylnicotinamide (114)

[0739] To a solution of 113 (80 mg, 0.33 mmol, 1.0 eq) in THF (3.0 mL) and H2O (1.0 mL) at 0°C was added NBS (62 mg, 0.35 mmol, 1.05 eq). The reaction mixture was stirred at 0°C for 0.5 h. The reaction was quenched with NaHCO3 and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using Hep / EtOAc (8 / 2) as an eluent to give 114 (87 mg, 0.30 mmol, 90%) as a pale yellow solid.

[0740] 1 H NMR (500 MHz, CDCl3) δ 8.25 (d,J= 7.4 Hz, 1H), 8.10 (d,J= 7.2 Hz, 1H), 6.47 (s, 1H), 4.80 (s, 2H), 3.10 (d,J= 4.2 Hz, 3H). ESI-MS m / z 290.1 ​​[M+H + ]

[0741] Step 5) Preparation of tert-butyl (S)-(1'-(5-((8-chloro-2-(6-chloro-5-(methylcarbamoyl)pyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (115)

[0742] To a solution of 89 (35 mg, 0.061 mmol, 1.0 eq) in acetonitrile (5.0 mL) at room temperature were added 114 (36 mg, 0.122 mmol, 2.0 eq) and NaHCO3 (20 mg, 0.244 mmol, 4.0 eq). The reaction mixture was stirred at 80°C for 16 h. The reaction was quenched with water and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give 115 (23 mg, 0.030 mmol, 49%) as a yellow solid.

[0743] 1 H NMR (500 MHz, MeOD) δ 8.50-8.44 (m, 1H), 8.33-8.27 (m, 2H), 8.20 (d,J= 7.8 Hz, 1H), 7.97 (d,J= 7.9 Hz, 1H), 7.26-7.17 (m, 4H), 7.05 (d,J= 9.8 Hz, 1H), 6.68 (d,J= 7.2 Hz, 1H), 4.93 (d,J= 9.9 Hz, 1H), 4.67 (s, 2H), 3.81-3.66 (m, 2H), 3.31-3.24 (m, 2H), 3.12 (d,J= 15.8 Hz, 1H), 2.95 (d,J= 11.7 Hz, 3H), 2.81 (d,J= 15.6 Hz, 1H), 2.00-1.92 (m, 1H), 1.88-1.81 (m, 1H), 1.75-1.69 (m, 1H), 1.54-1.47 (m, 10H). ESI-MS m / z 761.1 [M+H + ]

[0744] Step 6) Preparation of (S)-6-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-2-chloro-N-methylnicotinamide (Compound 53)

[0745] To a solution of 115 (23 mg, 0.030 mmol, 1.0 eq) in DCM (5.0 mL) at 0°C was added 4N-HCl (0.08 mL, 0.30 mmol, 10 eq) in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 h and the solvent was evaporated in vacuo. The reaction was quenched with saturated aqueous NaHCO3 solution (1.0 mL), diluted with water (5.0 mL), and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give compound 53 (4.4 mg, 0.007 mmol, 20%) as a pale yellow solid.

[0746] 1 H NMR (400 MHz, MeOD) δ 8.49 (s, 1H), 8.36-8.28 (m, 2H), 8.22 (d,J= 7.9 Hz, 1H), 7.98 (d,J= 7.8 Hz, 1H), 7.45-7.37 (m, 1H), 7.29-7.19 (m, 3H), 6.71 (d,J= 7.2 Hz, 1H), 4.69 (s, 2H), 4.06 (s, 1H), 3.84-3.74 (m, 2H), 3.30-3.20 (m, 2H), 3.15 (d,J= 15.7 Hz, 1H), 2.96 (s, 3H), 2.86 (d,J= 15.9 Hz, 1H), 2.05-1.89 (m, 3H), 1.64 (d,J= 12.6 Hz, 1H), 1.52 (d,J= 14.3 Hz, 1H). ESI-MS m / z 661.1 [M+H + ]

[0747]

[0748] Compound 54. tert-Butyl (S)-(1'-(5-((8-chloro-2-(6-(methylcarbamoyl)pyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate

[0749] Compound 55. (S)-6-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-N-methylpicolinamide

[0750] Step 1) Preparation of 6-bromo-N-methylpicolinamide (116)

[0751] To a solution of 6-bromopicolinic acid (300 mg, 1.49 mmol, 1.0 eq) in DMF (2.0 mL) were added methylamine HCl salt (131 mg, 1.94 mmol, 1.3 eq), HATU (850 mg, 2.24 mmol, 1.5 eq), and DIEA (0.78 mL, 4.47 mmol, 3.0 eq) at room temperature. The reaction mixture was stirred at room temperature for 16 h. The reaction was quenched with water, brine, and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using Hep / EtOAc (1 / 1) as an eluent to give 116 (211 mg, 0.98 mmol, 66%) as a yellow solid.

[0752] ESI-MS m / z 214.1 [M+H + ]

[0753] Step 2) Preparation of 6-(2-bromoacetyl)-N-methylpicolinamide (117)

[0754] To a solution of 116 (211 mg, 0.98 mmol, 1.0 eq) in dioxane (5.0 mL) at room temperature were added Pd(PPh3)2Cl2 (69 mg, 0.098 mmol, 0.1 eq), CuI (196 mg, 1.03 mmol, 0.15 eq), and tributyl(1-ethoxyvinyl)stannane (0.36 mL, 1.08 mmol, 1.1 eq). After stirring at 80 °C for 3 h, the reaction mixture was cooled to room temperature, quenched with water (5.0 mL), and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4. The solvent was removed in vacuo. The concentrate was briefly purified by silica gel column chromatography using Hep / EtOAc (1 / 1) as the eluent to give 6-(1-ethoxyvinyl)-N-methylpicolinamide (160 mg, 0.78 mmol, 79%) as a yellow oil. The unstable product was used in the next reaction without further purification.

[0755] To a solution of 6-(1-ethoxyvinyl)-N-methylpicolinamide (150 mg, 0.73 mmol, 1.0 eq) in THF (3.0 mL) and H2O (1.0 mL) was added NBS (136 mg, 0.76 mmol, 1.05 eq) at room temperature. The reaction mixture was stirred at room temperature for 1 h. The reaction was quenched with saturated aqueous NaHCO3 solution (5.0 mL) and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using Hep / EtOAc (8 / 2) as the eluent to give 117 (160 mg, 0.62 mmol, 86%) as a pale yellow solid.

[0756] 1H NMR (400 MHz, CDCl3) δ 8.47 (d,J= 7.5 Hz, 1H), 8.25 (d,J= 7.7 Hz, 1H), 8.09 (t,J= 7.7 Hz, 1H), 7.81 (s, 1H), 4.81 (s, 2H), 3.13 (d,J= 5.0 Hz, 3H). ESI-MS m / z 256.1 [M+H + ]

[0757] Step 3) Preparation of tert-butyl (S)-(1'-(5-((8-chloro-2-(6-(methylcarbamoyl)pyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (Compound 54)

[0758] To a solution of 89 (35 mg, 0.061 mmol, 1.0 eq) in acetonitrile (5.0 mL) at room temperature were added 117 (31 mg, 0.122 mmol, 2.0 eq) and NaHCO3 (20 mg, 0.244 mmol, 4.0 eq). The reaction mixture was stirred at 80 °C for 16 h. The reaction was quenched with water and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4, and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give compound 54 (20 mg, 0.027 mmol, 45%) as a yellow solid.

[0759] 1H NMR (400 MHz, MeOD) δ 8.66 (s, 1H), 8.33-8.28 (m, 2H), 8.26-8.20 (m, 1H), 8.08-8.03 (m, 2H), 7.23 (s, 4H), 6.72 (d,J= 7.2 Hz, 1H), 4.68 (s, 2H), 4.61 (s, 2H), 3.81-3.67 (m, 2H), 3.30-3.23 (m, 1H), 3.08-3.04 (m, 3H), 2.82 (d,J= 15.3 Hz, 1H), 2.07-2.03 (m, 1H), 2.00-1.93 (m, 1H), 1.89-1.80 (m, 1H), 1.76-1.69 (m, 1H), 1.53-1.45 (m, 10H). ESI-MS m / z 727.1 [M+H + ]

[0760] Step 4) Preparation of (S)-6-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-N-methylpicolinamide (Compound 55)

[0761] To a solution of compound 54 (18 mg, 0.025 mmol, 1.0 eq) in DCM (5.0 mL) at 0°C was added 4N-HCl (0.06 mL, 0.25 mmol, 10 eq) in 1,4-dioxane. The resulting mixture was stirred at room temperature for 3 h and the solvent was evaporated in vacuo. The residue was diluted with saturated aqueous NaHCO3 solution (5 mL) and extracted with EtOAc (2 X 25 mL). The combined organic layers were dried over Na2SO4 and the solvent was removed in vacuo. The concentrate was purified by silica gel column chromatography using DCM / MeOH (9 / 1) as an eluent to give compound 55 (2.5 mg, 0.004 mmol, 16%) as a pale yellow solid.

[0762] 1H NMR (400 MHz, MeOD) δ 8.66 (s, 1H), 8.34-8.28 (m, 2H), 8.26-8.21 (m, 1H), 8.08-8.01 (m, 2H), 7.43-7.37 (m, 1H), 7.27-7.19 (m, 3H), 6.73 (d,J= 7.2 Hz, 1H), 4.69 (s, 2H), 4.03 (s, 1H), 3.79 (d,J= 13.4 Hz, 2H), 3.30-3.21 (m, 2H), 3.15 (d,J= 15.7 Hz, 1H), 3.05 (s, 3H), 2.84 (d,J= 15.7 Hz, 1H), 2.02-1.89 (m, 2H), 1.64 (d,J= 13.8 Hz, 1H), 1.50 (d,J= 13.4 Hz, 1H). ESI-MS m / z 623.1 [M+H + ]

[0763] <Experimental Example 1: SHP2 phosphatase assay (IC 50 )>

[0764] SHP2 is allosterically activated by the binding of a bis-tyrosyl-phosphorylated peptide to its Src homology 2 (SH2) domain. This activation results in the release of the autoinhibitory interface of SHP2, which ultimately activates the SHP2 protein tyrosine phosphatase (PTP), making it available for substrate recognition and reaction catalysis. SHP2 catalytic activity was monitored using DiFUMP, a surrogate, in a rapid fluorescence-based assay.

[0765] The compound of the present invention was diluted to an appropriate multiple using 100% DMSO and analysis buffer, and the final test concentration of the compound of the present invention was used by diluting it to three times from 10 μM or 1 μM.

[0766] SHP2 enzyme activity assays were performed at room temperature in 96-well black polystyrene plates (flat bottom, non-binding surface) with a final reaction volume of 100 μL and the following measurement and buffer conditions. The buffer conditions were 60 mM HEPES, 75 mM NaCl, 75 mM KCl, 1 mM EDTA, 0.05% Tween 20, and 2 mM DTT. PTPs were activated by incubating the above buffer with the SHP2 activating peptide (dPEG8) for 30 minutes. DMSO or a compound of the present invention was added to the mixture, and the reaction was incubated at room temperature for 1 hour. After that, the substrate DiFUMP (Invitrogen, Cat#D6567) was added to the reaction at room temperature for 1 hour. The fluorescence signal was measured with an enzyme reader (Envision, Perkin-Elmer) using an excitation and emission wavelength of 340 nM and 450 nM, respectively.

[0767] The following calculation formula was used for data analysis.

[0768] Inhibition rate % = [1-(conversion rate_ 샘플 - Conversion rate_ min ) / (Conversion rate_ max - Conversion rate_ min )] X 100%

[0769] Here, the conversion rate_ 샘플 is the average reading of the sample well; conversion rate_ min is the average of the blank control wells, representing the readings of wells with no enzyme activity; Conversion rate_ max is the average value of the positive control wells, representing the reading of the wells without inhibition.

[0770] The dose-response curve was fitted in GraphPad Prism software, and the IC was calculated using the “log[inhibitor] vs. response-variable slope” program. 50 Calculate the value.

[0771] Phosphatase IC using the compound according to the present invention as shown in Table 1 below 50 The activity results were confirmed by measuring the values.

[0772] Example Compound Structural Formula PTPN11 / SHP2-FL IC 50 Compound 1 (comparative substance) ++Compound 2 +++Compound 3 (comparative substance) +Compound 4 ++Compound 5 ++Compound 6 +++Compound 7 +++Compound 8 +++Compound 9 ++Compound 10 ++Compound 11 ++Compound 12 ++Compound 13 +++Compound 14 +++Compound 15 +++Compound 16 +++Compound 17 +++Compound 18 +++Compound 19 +++Compound 20 +++Compound 21 +++Compound 22 +++Compound 23 +++Compound 24 +++Compound 25 +++Compound 26 +++Compound 27 +++Compound 28 +++Compound 29 +++Compound 30 +++Compound 31 +++Compound 32 +++Compound 33 +++Compound 34 +++Compound 35 +++Compound 36 +++Compound 37 +++Compound 38 +++Compound 39 +++Compound 40 +++Compound 41 +++Compound 42 ++Compound 43 +++Compound 44 +++Compound 45 +++Compound 46 +++Compound 47 +++Compound 48 ++Compound 49 ++Compound 50 +++Compound 51 +++Compound 52 +++Compound 53 +++Compound 54 NA compound 55 +++PTPN11 / SHP2-FL IC 50 Inhibitory activity:+: 1–100 μM IC 50 values ​​++: 100 nM~1 μM IC 50 values ​​+++: <100 nM IC 50 valuesNA: Not Active

[0773] As shown in Table 1, the compounds of the present invention have excellent phosphatase IC 50 It has a value. In particular, through a structure-activity comparison of the compounds of the present invention, in the chemical formula 1 of the present invention, R1 is hydrogen; R3 is halogen; R4 is hydrogen or C1-C6 alkyl; R5 is substituted or unsubstituted 2-oxa-8-azaspiro[4.5]decan-8-yl or substituted or unsubstituted 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl; R6 is hydroxymethyl, C1-C6 alkoxymethyl or C1-C6 alkoxymethoxymethyl; Compounds in which R2 is substituted with hydrogen, C1-C6 alkyl or C1-C6 alkoxyC1-C6 alkyl (Compounds 1, 3, 4, 9 to 12) are preferred over compounds in which R2 is substituted with C1-C6 alkylC(=O)NHC1-C6 alkyl-, C1-C6 alkoxyC1-C6 alkylNHC(=O)-,

[0774] , , Excellent phosphatase IC in compounds substituted with 6-10 membered aryl, 5-10 membered heteroaryl, or 4-10 membered heterocycloalkyl (compounds 2, 5, 6-8, 13-53, 55) 50 It was confirmed that the value and excellent anticancer activity were obtained from this.

[0775] <Experimental Example 2: Cell viability analysis (GI 50 )>

[0776] To confirm the cell viability by the synthesized compound, NCI-H358 cells (3000 cells / well) were seeded in 96-well white plates and cultured for 24 hours in a 37°C, 5% CO2 incubator. After 24 hours, the cells were treated with DMSO or various concentrations (9 concentrations, 3-fold dilution from 30 μM) diluted in RPMI 1640 medium and cultured for 5 days in a 37°C, 5% CO2 incubator.

[0777] After equilibration of the cultured cells at room temperature for 30 minutes, cell viability was evaluated using the Cell-Titer glo assay according to the manufacturer's protocol. Briefly, the same amount of CellTiter-Glo luminescent cell activity assay solution was added to each well where cells were cultured, the cells were gently shaken for 2 minutes, and incubated at room temperature for 10 minutes. Luminescence was measured using a multifunctional microplate reader (GloMAx Discover, Promega). Cell viability was calculated by analyzing the measured luminescence results, and then GI was determined through nonlinear regression analysis using GraphPad Prism software. 50 The values ​​were derived. Cell viability was expressed as a percentage of absorbance.

[0778] Cell viability (%) = (luminescence of sample treatment group / luminescence of DMSO treatment group) × 100

[0779] Using the compound according to the present invention, cell viability GI is as shown in Table 2 below. 50 The activity results were confirmed by measuring the values.

[0780] Example GI 50 _NCI-H358 Example GI 50 _NCI-H358 Example GI 50 _NCI-H358 Compound 1+ Compound 20+ Compound 39+++ Compound 2+ Compound 21++ Compound 40++ Compound 3 NT Compound 22+ Compound 41++ Compound 4+ Compound 23+ Compound 42++ Compound 5+ Compound 24++ Compound 43++ Compound 6+ Compound 25++ Compound 44+++ Compound 7++ Compound 26+ Compound 45+++ Compound 8++ Compound 27++ Compound 46+++ Compound 9 NT Compound 28+ Compound 47++ Compound 10 NT Compound 29++ Compound 48++ Compound 11 NT Compound 30++ Compound 49+++ Compound 12NT Compound 31++ Compound 50++ Compound 13+ Compound 32++ Compound 51++ Compound 14++ Compound 33++ Compound 52++ Compound 15++ Compound 34++ Compound 53+++ Compound 16+ Compound 35++ Compound 53+++ Compound 17++ Compound 36++ Compound 54NT Compound 18+ Compound 37++ Compound 55+++ Compound 19+ Compound 38++ NCI-H358GI 50 + : 1~100 uM GI 50 values++ :100~1000 nM GI 50 values+++ : <100 nM GI 50 valuesNT: Not tested

[0781] As shown in Table 2, the compounds of the present invention have excellent cell viability GI 50It has a value. In the chemical formula 1 of the present invention, R1 is hydrogen; R3 is halogen; R4 is hydrogen or C1-C6 alkyl; R5 is substituted or unsubstituted 2-oxa-8-azaspiro[4.5]decan-8-yl or substituted or unsubstituted 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl; R6 is hydroxymethyl, C1-C6 alkoxymethyl or C1-C6 alkoxymethoxymethyl; and R2 is C1-C6 alkylC(=O)NHC1-C6 alkyl-, C1-C6 alkoxyC1-C6 alkylNHC(=O)-, rather than compounds in which the R2 substituent is substituted with hydrogen, C1-C6 alkyl or C1-C6 alkoxyC1-C6 alkyl (Compounds 1, 3, 4, 9 to 12). , , , 6-10 membered ring aryl, 5-10 membered ring heteroaryl, or 4-10 membered ring heterocycloalkyl substituted compounds (compounds 2, 5, 6-8, 13-53, 55) phosphatase IC of Table 1 50 Excellent cell viability GI similar to the value 50 The values ​​were shown to confirm excellent anticancer activity.

[0782] <Example 1. Preparation of a powder>

[0783] 2 g of the compound of the present invention and 1 g of lactose were mixed and filled into a sealed bag to prepare a powder.

[0784] <Example 2. Preparation of tablets>

[0785] 100 mg of the compound of the present invention 2, 100 mg of microcrystalline cellulose, 60 mg of lactose monohydrate, 20 mg of low-substituted hydroxypropyl cellulose, and 2 mg of magnesium stearate were mixed and then compressed into tablets according to a conventional tablet manufacturing method.

[0786] <Example 3. Preparation of capsules>

[0787] 100 mg of the compound 2 of the present invention, 100 mg of microcrystalline cellulose, 60 mg of lactose monohydrate, 20 mg of low-substituted hydroxypropyl cellulose, and 2 mg of magnesium stearate were mixed, and then the above ingredients were mixed according to a conventional capsule manufacturing method and filled into a gelatin capsule to manufacture a capsule.

[0788] <Example 4. Preparation of injection>

[0789] After mixing 10 mg of the compound of the present invention, an appropriate amount of sterile distilled water for injection, and an appropriate amount of a pH regulator, the mixture was prepared with the above ingredient contents per 1 ampoule (2 ml) according to a conventional method for preparing injections.

Claims

1. A compound having the following chemical formula 1, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof; [Chemical Formula 1] Among the above formulas, R 1 Silver hydrogen, C 1 -C 6 Alkoxy C 1 -C 6 AlkylNHC(=O)-, , or and; R 2 is C 1 -C 6 Alkoxy, C 1 -C 6 Alkoxy C 1 -C 6 Alkyl, C 1 -C 6 AlkylC(=O)NHC 1 -C 6 Alkyl-, C 1 -C 6 Alkoxy C 1 -C 6 AlkylNHC(=O)-, , , or And, In the above formula, A is a substituted or unsubstituted 6-10 membered aryl, a substituted or unsubstituted 5-10 membered heteroaryl, or a substituted or unsubstituted 4-10 membered heterocycloalkyl; R 3 is a halogen; R 4 is hydrogen or C 1 -C 6 It is alkyl; R 5 is substituted or unsubstituted 2-oxa-8-azaspiro[4.5]decan-8-yl or substituted or unsubstituted 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl, wherein the substituted 2-oxa-8-azaspiro[4.5]decan-8-yl or 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl is each independently hydrogen, amino and C 1 -C 6 Substituted with one or more substituents selected from the group consisting of alkyl; R 6 Silver hydroxymethyl, C 1 -C 6 Alkoxymethyl or C 1 -C 6 It is alkoxymethoxymethyl; R 7 are each independently hydrogen, halogen, amino, cyano, acetyl, carboxyl, C 1 -C 6 Alkyl, HaloC 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Alkoxy-C(=O)-, C 1 -C 6 Alkyl-C(=O)-, C 1 -C 6 Alkyl-C(=O)NH-, C 1 -C 6 AlkylNH-, C 1 -C 6 Alkyl-NHC(=O)-, C 1 -C 6 Alkyl-OC(=O)NH-, C 1 -C 6 Alkyl-SO 2 NH-, C 1 -C 6 Alkoxy-C 1 -C 6 Alkyl-NHC(=O)-(C 1 -C 6 Alkoxy) 0-1 -, C 1 -C 6 Alkyl-NHC(=O)-(C 1 -C 6 Alkoxy) 0-1 -, COOH-(C 1 -C 6 Alkoxy) 0-1 -, , , or and; m is 0, 1 or 2; 2. A compound having the following chemical formula 2, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof; [Chemical formula 2] Among the above formulas, R 1 is hydrogen; R 2 is C 1 -C 6 AlkylC(=O)NHC 1 -C 6 Alkyl-, C 1 -C 6 Alkoxy C 1 -C 6 AlkylNHC(=O)-, , , or And, If A in the above formula is phenyl or pyridinyl, R 7 are each independently hydrogen, halogen, amino, cyano, acetyl, carboxyl, C 1 -C 6 Alkyl, HaloC 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Alkoxy-C(=O)-, C 1 -C 6 Alkyl-C(=O)-, C 1 -C 6 Alkyl-C(=O)NH-, C 1 -C 6 AlkylNH-, C 1 -C 6 Alkyl-NHC(=O)-, C 1 -C 6 Alkyl-OC(=O)NH-, C 1 -C 6 Alkyl-SO 2 NH-, C 1 -C 6 Alkoxy-C 1 -C 6 Alkyl-NHC(=O)-(C 1 -C 6 Alkoxy) 0-1 -, C 1 -C 6 Alkyl-NHC(=O)-(C 1 -C 6 Alkoxy) 0-1 -, COOH-(C 1 -C 6 Alkoxy) 0-1 -, , , or and; In the above formula, if A is pyrrolyl, furanyl, thiophenyl or pyrimidinyl, R 7 are each independently hydrogen, halogen, amino, cyano, acetyl, carboxyl, C 1 -C 6 Alkyl, HaloC 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Alkoxy-C(=O)-, C 1 -C 6 Alkyl-C(=O)-, C 1 -C 6 Alkyl-C(=O)NH-, C 1 -C 6 AlkylNH-, C 1 -C 6 Alkyl-NHC(=O)-, C 1 -C 6 Alkyl-OC(=O)NH-, C 1 -C 6 Alkyl-SO 2 NH-, C 1 -C 6 Alkoxy-C 1 -C 6 Alkyl-NHC(=O)-(C 1 -C 6 Alkoxy) 0-1 -, C 1 -C 6 Alkyl-NHC(=O)-(C 1 -C 6 Alkoxy) 0-1 -, or COOH-(C 1 -C 6 Alkoxy) 0-1 - and; In the above formula, if A is piperidinyl, R 7 are each independently hydrogen, halogen, amino, cyano, acetyl, carboxyl, C 1 -C 6 Alkyl, HaloC 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Alkoxy-C(=O)-, C 1 -C 6 Alkyl-C(=O)-, C 1 -C 6 Alkyl-C(=O)NH-, C 1 -C 6 AlkylNH-, C 1 -C 6 Alkyl-NHC(=O)-, C 1 -C 6 Alkyl-OC(=O)NH-, C 1 -C 6 Alkyl-SO 2 NH-, C 1 -C 6 Alkoxy-C 1 -C 6 Alkyl-NHC(=O)-(C 1 -C 6 Alkoxy) 0-1 -, C 1 -C 6 Alkyl-NHC(=O)-(C 1 -C 6 Alkoxy) 0-1 -, or COOH-(C 1 -C 6 Alkoxy) 0-1 - and; R 3 is a halogen; R 4 is hydrogen or C 1 -C 6 It is alkyl; R 5 is substituted or unsubstituted 2-oxa-8-azaspiro[4.5]decan-8-yl or substituted or unsubstituted 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl, wherein the substituted 2-oxa-8-azaspiro[4.5]decan-8-yl or 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl is each independently hydrogen, amino and C 1 -C 6 Substituted with one or more substituents selected from the group consisting of alkyl; R 6 Silver hydroxymethyl, C 1 -C 6 Alkoxymethyl or C 1 -C 6 It is alkoxymethoxymethyl; m is 0, 1 or 2; 3. In paragraph 2, In the above chemical formula 2, Among the above formulas, R 1 is hydrogen; R 2 Is And, If A in the above formula is phenyl or pyridinyl, R 7 are each independently hydrogen, halogen, amino, cyano, acetyl, carboxyl, C 1 -C 6 Alkyl, HaloC 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Alkoxy-C(=O)-, C 1 -C 6 Alkyl-C(=O)-, C 1 -C 6 Alkyl-C(=O)NH-, C 1 -C 6 AlkylNH-, C 1 -C 6 Alkyl-NHC(=O)-, C 1 -C 6 Alkyl-OC(=O)NH-, C 1 -C 6 Alkyl-SO 2 NH-, C 1 -C 6 Alkoxy-C 1 -C 6 Alkyl-NHC(=O)-(C 1 -C 6 Alkoxy) 0-1 -, C 1 -C 6 Alkyl-NHC(=O)-(C 1 -C 6 Alkoxy) 0-1 -, COOH-(C 1 -C 6 Alkoxy) 0-1 -, , , or and; In the above formula, if A is pyrrolyl, furanyl, thiophenyl or pyrimidinyl, R 7 are each independently hydrogen, halogen, amino, cyano, acetyl, carboxyl, C 1 -C 6 Alkyl, HaloC 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Alkoxy-C(=O)-, C 1 -C 6 Alkyl-C(=O)-, C 1 -C 6 Alkyl-C(=O)NH-, C 1 -C 6 AlkylNH-, C 1 -C 6 Alkyl-NHC(=O)-, C 1 -C 6 Alkyl-OC(=O)NH-, C 1 -C 6 Alkyl-SO 2 NH-, C 1 -C 6 Alkoxy-C 1 -C 6 Alkyl-NHC(=O)-(C 1 -C 6 Alkoxy) 0-1 -, C 1 -C 6 Alkyl-NHC(=O)-(C 1 -C 6 Alkoxy) 0-1 -, or COOH-(C 1 -C 6 Alkoxy) 0-1 - and; In the above formula, if A is piperidinyl, R 7 are each independently hydrogen, halogen, amino, cyano, acetyl, carboxyl, C 1 -C 6 Alkyl, HaloC 1 -C 6 Alkyl, C 1 -C 6 Alkoxy, C 1 -C 6 Alkoxy-C(=O)-, C 1 -C 6 Alkyl-C(=O)-, C 1 -C 6 Alkyl-C(=O)NH-, C 1 -C 6 AlkylNH-, C 1 -C 6 Alkyl-NHC(=O)-, C 1 -C 6 Alkyl-OC(=O)NH-, C 1 -C 6 Alkyl-SO 2 NH-, C 1 -C 6 Alkoxy-C 1 -C 6 Alkyl-NHC(=O)-(C 1 -C 6 Alkoxy) 0-1 -, C 1 -C 6 Alkyl-NHC(=O)-(C 1 -C 6 Alkoxy) 0-1 -, or COOH-(C 1 -C 6 Alkoxy) 0-1 - and; R 3 is a halogen; R 4 is hydrogen or C 1 -C 6 It is alkyl; R 5 is substituted or unsubstituted 2-oxa-8-azaspiro[4.5]decan-8-yl or substituted or unsubstituted 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl, wherein the substituted 2-oxa-8-azaspiro[4.5]decan-8-yl or 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl is each independently hydrogen, amino and C 1 -C 6 Substituted with one or more substituents selected from the group consisting of alkyl; R 6 Silver hydroxymethyl, C 1 -C 6 Alkoxymethyl or C 1 -C 6 It is alkoxymethoxymethyl; m is 0, 1 or 2; 4. In paragraph 3, In the above chemical formula 2, Among the above formulas, R 5 is a substituted or unsubstituted 1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl, Here, the substituted 1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl is each independently hydrogen, amino and C 1 -C 6 It is substituted with one or more substituents selected from the group consisting of alkyl.

5. In paragraph 3 or 4, In the above chemical formula 2 R 3 is chloro; R 4 is hydrogen or methyl; 6. A compound characterized by being any one selected from the group of compounds below, or a pharmaceutically acceptable salt thereof; (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(4-methoxyphenyl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 2); (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(methoxymethyl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 4); N-((7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)methyl)acetamide hydrochloride (Compound 5); (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(pyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 6); (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(1H-pyrrol-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 7); (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(thiophen-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 8); (7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-2-methylimidazo[1,2-a]pyridin-3-yl)(piperazin-1-yl)methanone dihydrochloride (Compound 9); (7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-2-methylimidazo[1,2-a]pyridin-3-yl)(4-aminopiperidin-1-yl)methanone dihydrochloride (Compound 10); 7-((5-((3S,4S)-4-Amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-N-(2-methoxyethyl)-2-methylimidazo[1,2-a]pyridine-3-carboxamide hydrochloride (Compound 11); 7-((5-((3S,4S)-4-Amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-2-methyl-N-(piperidin-4-yl)imidazo[1,2-a]pyridine-3-carboxamide dihydrochloride (Compound 12); (7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)(piperazin-1-yl)methanone dihydrochloride (Compound 13); (7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)(4-aminopiperidin-1-yl)methanone dihydrochloride (Compound 14); 7-((5-((3S,4S)-4-Amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-N-(piperidin-4-yl)imidazo[1,2-a]pyridine-2-carboxamide dihydrochloride (Compound 15); 7-((5-((3S,4S)-4-Amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-N-(2-methoxyethyl)imidazo[1,2-a]pyridine-2-carboxamide hydrochloride (Compound 16); 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(piperazin-1-yl)ethanone dihydrochloride (Compound 17); 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(4-aminopiperidin-1-yl)ethanone dihydrochloride (Compound 18); 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(piperidin-4-yl)acetamide dihydrochloride (Compound 19); 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(2-methoxyethyl)acetamide hydrochloride (Compound 20); 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-methylacetamide hydrochloride (Compound 21); 2-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetic acid hydrochloride (Compound 22); (S)-2-(3-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetic acid hydrochloride (Compound 23); 2-(3-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(piperazin-1-yl)ethan-1-one dihydrochloride (Compound 24); 2-(3-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(2-methoxyethyl)acetamide hydrochloride (Compound 25); 4-(7-((5-((3S,4S)-4-Amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)benzoic acid hydrochloride (Compound 26); (4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenyl)(piperazin-1-yl)methanone dihydrochloride (Compound 27); 4-(7-((5-((3S,4S)-4-Amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-N-(2-methoxyethyl)benzamide hydrochloride (Compound 28); (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((2-(4-aminophenyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 29); N-(4-(7-((5-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenyl)acetamide hydrochloride (Compound 30); (S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(4-methoxyphenyl))imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 31); (S)-7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloro-N-(2-methoxyethyl)imidazo[1,2-a]pyridine-2-carboxamide hydrochloride (Compound 32); (S)-2-(4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)acetic acid hydrochloride (Compound 33); (S)-2-(4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(piperazin-1-yl)ethanone dihydrochloride (Compound 34); (S)-2-(4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-1-(4-aminopiperidin-1-yl)ethan-1-one dihydrochloride (Compound 35); (S)-2-(4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(piperidin-4-yl)acetamide dihydrochloride (Compound 36); (S)-2-(4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-(hydroxymethyl)-3-methylpyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenoxy)-N-(2-methoxyethyl)acetamide hydrochloride (Compound 37); (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(4-methoxyphenyl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol (Compound 38); (S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(4-methoxyphenyl))imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol (Compound 39); (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(furan-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 40); (3-((3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl)-6-((8-chloro-2-(6-chloropyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-5-methylpyrazin-2-yl)methanol hydrochloride (Compound 41); (S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(6-methoxypyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol hydrochloride (Compound 42); tert-Butyl ((3S,4S)-8(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(piperidin-4-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol dihydrochloride (Compound 43); (S)-1-(4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)piperidin-1-yl)ethan-1-one (Compound 44); Ethyl (S)-4-(7-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)piperidine-1-carboxylate (Compound 45); (S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(5-methoxypyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol hydrochloride (Compound 46); (S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((8-chloro-2-(2-methoxypyrimidin-4-yl)imidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol hydrochloride (Compound 47); (S)-4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-N-(2-methoxyethyl)benzamide (Compound 48); (S)-4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-N-methylbenzamide (Compound 49); (S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((2-(4-aminophenyl)-8-chloroimidazo[1,2-a]pyridin-7-yl)thio)pyrazin-2-yl)methanol (Compound 50); (S)-N-(4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)phenyl)acetamide (Compound 51); (S)-4-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-2-fluoro-N-methylbenzamide hydrochloride (Compound 52); (S)-6-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-2-chloro-N-methylnicotinamide (Compound 53); tert-Butyl (S)-(1'-(5-((8-chloro-2-(6-(methylcarbamoyl)pyridin-2-yl)imidazo[1,2-a]pyridin-7-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)carbamate (Compound 54); and (S)-6-(7-((5-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)-8-chloroimidazo[1,2-a]pyridin-2-yl)-N-methylpicolinamide (Compound 55).

7. A pharmaceutical composition for preventing or treating cancer, comprising a therapeutically effective amount of a compound of any one of claims 1 to 6, a stereoisomer thereof, a solvate thereof, a prodrug thereof, or a pharmaceutically acceptable salt thereof.

8. In paragraph 7, A pharmaceutical composition, characterized in that the cancer is a solid cancer or a blood cancer.

9. In paragraph 8, The above solid cancers include brain cancer, brain tumor, benign astrocytoma, malignant astrocytoma, pituitary adenoma, meningioma, brain lymphoma, oligodendroglioma, intracranial tumor, ependymoma, brainstem tumor, head and neck tumor, glioma, glioblastoma, laryngeal cancer, oropharyngeal cancer, nasal cancer, nasopharyngeal cancer, salivary gland cancer, hypopharyngeal cancer, tonsil cancer, thyroid cancer, oral cancer, esophageal cancer, eye cancer, thoracic tumor, small cell lung cancer, non-small cell lung cancer, thymic cancer, lung adenocarcinoma, lung cancer, lung squamous cell carcinoma, pleural cancer, mediastinal tumor, breast cancer, male breast cancer, abdominal tumor, stomach cancer, gastric mastoid tumor, liver cancer, hepatoblastoma, gallbladder cancer, biliary tract cancer, pancreatic cancer, small intestine cancer, colon cancer, colon cancer, rectal cancer, duodenal cancer, anal cancer, urinary bladder cancer, kidney cancer, renal pelvis cancer, heart cancer, peritoneal cancer, A pharmaceutical composition characterized in that the cancer is adrenal cancer, spinal cancer, bone cancer, genital tumor, penile cancer, prostate cancer, female genital tumor, cervical cancer, endometrial cancer, ovarian cancer, uterine sarcoma, vaginal cancer, female external genital cancer, female urethral cancer or skin cancer.

10. In paragraph 8, A pharmaceutical composition characterized in that the blood cancer is acute myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, childhood lymphoma, malignant lymphoma, multiple myeloma, or aplastic anemia.

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