Pyrimidines for degrading bruton's tyrosine kinase

Abstract

This invention provides compounds of formula (I). ; Where R 1. R 2. R 3. R 4A, R 4b and R 5 series as defined herein; and its pharmaceutically acceptable salts, which can be used as a treatment for CLL.

Description

[Previous Technology]

[0001] Bruton's tyrosine kinase (Btk) is a key signal transduction enzyme manifested in hematopoietic cell types. Btk plays an essential role in the B-cell signaling pathway, which links stimulation of the cell surface B-cell receptor (BCR) to downstream intracellular responses. Btk is a key regulator of B-cell development, activation, signal transduction, and survival (Kurosaki, Curr Op Imm, 2000, 276-281; ​​Schaeffer and Schwartzberg, Curr Op Imm 2000, 282-288). In addition, Btk plays a role in other hematopoietic cell signaling pathways, such as TNF-α production mediated by Toll-like receptor (TLR) and intercytokine receptors in macrophages, IgE receptor (FcεRI) signaling in mast cells, inhibition of Fas / APO-1 apoptosis signaling in B-lineage lymphoid cells, and collagen-stimulated platelet aggregation. See, for example, CA Jeffries, et al., (2003), Journal of Biological Chemistry 278:26258-26264; NJ Horwood, et al., (2003), The Journal of Experimental Medicine 197:1603-1611; Iwaki et al. (2005), Journal of Biological Chemistry 280(48):40261-40270; Vassilev et al. (1999), Journal of Biological Chemistry 274(3):1646-1656; and Quek et al. (1998), Current Biology 8(20):1137-1140. Currently, Btk inhibitors are under development. As an alternative to inhibition, the protein hydrolysis and degradation of Btk can affect B cell function not only by blocking BCR signaling but also by reducing the presence of Btk proteins from within the cell itself. A reduction in Btk protein will decrease Btk kinase activity and reduce any protein interactions or scaffold function of Btk. Therefore, there is a medical need to develop Btk degrading agents. [Summary of the Invention]

[0002] In some embodiments, the compounds provided herein induce protein hydrolytic degradation of Btk via the ubiquitin protein hydrolysis pathway.

[0003] In some embodiments, a compound of formula (I) is provided: (I) wherein R1 is selected from the group consisting of hydrogen, C1-C3 alkyl, CN, C1-C3 haloalkyl, halogen, CH2OCH3, CH2OH, and CH2CN; R2 and R3 are each independently selected from the group consisting of hydrogen and halogen; R4a and R4b are each independently selected from the group consisting of hydrogen, C1-C3 alkyl, and CH2OCH3; and R5 is selected from the group consisting of hydrogen, halogen, and C1-C3 alkyl; or a pharmaceutically acceptable salt thereof.

[0004] In some embodiments, R1 is selected from the group consisting of: hydrogen, C1-C3 alkyl, CN (cyano), C1-C3 haloalkyl, halo, CH2OCH3; CH2OH; and CH2CN, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is a C1-C3 alkyl, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is a methyl, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is hydrogen, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is CN, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is a C1-C3 haloalkyl, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is CF3, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is a halo, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is Cl, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is F, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is CH2OCH3, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is CH2OH, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is CH2CN, or a pharmaceutically acceptable salt thereof.

[0005] In some embodiments, R2 is selected from the group consisting of hydrogen and halogen groups or pharmaceutically acceptable salts thereof. In some embodiments, R2 is hydrogen or a pharmaceutically acceptable salt thereof. In some embodiments, R2 is a halogen group or a pharmaceutically acceptable salt thereof. In some embodiments, R2 is F or a pharmaceutically acceptable salt thereof.

[0006] In some embodiments, R3 is selected from the group consisting of hydrogen and halogen groups or pharmaceutically acceptable salts thereof. In some embodiments, R3 is hydrogen or a pharmaceutically acceptable salt thereof. In some embodiments, R3 is a halogen group or a pharmaceutically acceptable salt thereof. In some embodiments, R3 is F or a pharmaceutically acceptable salt thereof.

[0007] In some embodiments, R4a and R4b are each independently selected from the group consisting of hydrogen, C1-C3 alkyl, and CH2OCH3, or pharmaceutically acceptable salts thereof. In some embodiments, R4a is hydrogen and R4b is hydrogen, or a pharmaceutically acceptable salt thereof. In some embodiments, R4a is hydrogen and R4b is methyl, or a pharmaceutically acceptable salt thereof. In some embodiments, R4a is methyl and R4b is hydrogen, or a pharmaceutically acceptable salt thereof. In some embodiments, R4a is hydrogen and R4b is ethyl, or a pharmaceutically acceptable salt thereof. In some embodiments, R4a is ethyl and R4b is hydrogen, or a pharmaceutically acceptable salt thereof. In some embodiments, R4a is methyl and R4b is methyl, or a pharmaceutically acceptable salt thereof. In some embodiments, R4a is hydrogen and R4b is isopropyl, or a pharmaceutically acceptable salt thereof. In some embodiments, R4a is isopropyl and R4b is hydrogen, or a pharmaceutically acceptable salt thereof. In some embodiments, R4a is hydrogen and R4b is CH2OCH3, or a pharmaceutically acceptable salt thereof. In some embodiments, R4a is CH2OCH3 and R4b is hydrogen, or a pharmaceutically acceptable salt thereof.

[0008] In some embodiments, R5 is selected from the group consisting of hydrogen, halogen groups, and C1-C3 alkyl groups, or pharmaceutically acceptable salts thereof. In some embodiments, R5 is hydrogen, or a pharmaceutically acceptable salt thereof. In some embodiments, R5 is a halogen group, or a pharmaceutically acceptable salt thereof. In some embodiments, R5 is F, or a pharmaceutically acceptable salt thereof. In some embodiments, R5 is Cl, or a pharmaceutically acceptable salt thereof. In some embodiments, R5 is methyl, or a pharmaceutically acceptable salt thereof.

[0009] In some embodiments, R1 is methyl and R2 is hydrogen, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is methyl, R2 is hydrogen, and R3 is hydrogen, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is methyl, R2 is hydrogen, R3 is hydrogen, R4a is hydrogen, and R4b is methyl, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is methyl, R2 is hydrogen, R3 is hydrogen, and R5 is hydrogen, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is methyl, R4a is hydrogen, and R4b is methyl, or a pharmaceutically acceptable salt thereof. In some embodiments, R2 is hydrogen, R4a is hydrogen, and R4b is methyl, or a pharmaceutically acceptable salt thereof. In some embodiments, R3 is hydrogen, R4a is hydrogen, and R4b is methyl, or a pharmaceutically acceptable salt thereof.

[0010] In some embodiments, R5 is hydrogen, R4a is hydrogen, and R4b is methyl, or a pharmaceutically acceptable salt thereof. In some embodiments, R5 is hydrogen, R4a is hydrogen, R4b is methyl, and R3 is hydrogen, or a pharmaceutically acceptable salt thereof. In some embodiments, R5 is hydrogen, R4a is hydrogen, R4b is methyl, R3 is hydrogen, and R2 is hydrogen, or a pharmaceutically acceptable salt thereof. In some embodiments, R5 is hydrogen, R4a is hydrogen, R4b is methyl, R3 is hydrogen, and R1 is methyl, or a pharmaceutically acceptable salt thereof.

[0011] In some embodiments, R1 is methyl and R3 is a halogroup, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is methyl and R3 is F, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is methyl and R2 is a halogroup, or a pharmaceutically acceptable salt thereof. In some embodiments, R1 is methyl and R2 is F, or a pharmaceutically acceptable salt thereof.

[0012] In some embodiments, R5 is a halogroup, R4a is hydrogen, and R4b is methyl, or a pharmaceutically acceptable salt thereof. In some embodiments, R5 is F, R4a is hydrogen, and R4b is methyl, or a pharmaceutically acceptable salt thereof. In some embodiments, R5 is a halogroup, R4a is methyl, and R4b is hydrogen, or a pharmaceutically acceptable salt thereof. In some embodiments, R5 is F, R4a is methyl, and R4b is hydrogen, or a pharmaceutically acceptable salt thereof. In some embodiments, R5 is a halogroup, R4a is hydrogen, and R4b is ethyl, or a pharmaceutically acceptable salt thereof. In some embodiments, R5 is F, R4a is hydrogen, and R4b is ethyl, or a pharmaceutically acceptable salt thereof. In some embodiments, R5 is a halogroup, R4a is ethyl, and R4b is hydrogen, or a pharmaceutically acceptable salt thereof. In some embodiments, R5 is F, R4a is ethyl, and R4b is hydrogen, or a pharmaceutically acceptable salt thereof.

[0013] In some embodiments, pharmaceutical compositions are provided that comprise a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof in combination with a pharmaceutically acceptable carrier.

[0014] In some embodiments, pharmaceutical compositions are provided that comprise a therapeutically effective amount of, or a combination of, a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0015] In some embodiments, the compound is: or a pharmaceutically acceptable salt thereof.

[0016] In some embodiments, the compound system is:

[0017] In some embodiments, the compound is a pharmaceutically acceptable salt of the following:

[0018] In some embodiments, the compound is 3-tert-butyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazinan-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine, or a pharmaceutically acceptable salt thereof.

[0019] In some embodiments, the compound is 3-tert-butyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine.

[0020] In some embodiments, the compound is a pharmaceutically acceptable salt of the following: In some embodiments, the compound is 3-tert-butyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine.

[0021] In some embodiments, the compound is selected from the group consisting of: 3-tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine; 2 3-tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine; R)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine; 3 3-tertiary butyl-N-{(1 R)-1-[4-(6-{[5-(4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}piperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine; 4 3-tertiary butyl-N-[(1 R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)-3-fluorophenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine; 5 3-tertiary butyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)-3-fluorophenyl]ethyl]-1,2,4-diazol-5-methylamine; R)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)-3-fluorophenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine; 6 3-tertiary butyl-N-[(1 R)-1-{4-[6-({5-[(2 S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-ethylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine; 7 3-tertiary butyl- N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)-3-fluorophenyl]ethyl}-2-ethylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine;8 3-Tri-butyl-N-[(1R)-1-{4-[6-({5-[(2R)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-ethylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine; 9 3-Tri-butyl-N-{(1 R)-1-[4-(6-{[5-(4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2,2-dimethylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine; 10 3-tertiary butyl-N-[(1 R)-1-{4-[6-({5-[(2 S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-3-fluoro-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine; 11 3-Tributyl-N-{(1R)-1-[4-(6-{[5-(4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}piperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-3-fluoro-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine; 12 3-Tributyl- N-{(1R)-1-[4-(6-{[5-(4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}piperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-5-fluoro-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine; 13 3-tertiary butyl-N-[(1R)-1-{4-[6-({5-[(2 S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-(propyl-2-yl)piperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine; 14 3-Tributyl-N-[(1R)-1-{2-chloro-4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]phenyl}ethyl]-1,2,4-diazol-5-methylamine;15 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2R)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-(methoxymethyl)piperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine; 16 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2R)-4-{2-[ ... S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-trifluoromethyl)phenyl}ethyl]-1,2,4-diazol-5-methylamine; and 17,3-tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-fluorophenyl}ethyl]-1,2,4-diazol-5-methylamine; or a pharmaceutically acceptable salt thereof. Definition of terms;

[0022] As used in the specification and the accompanying claims, unless otherwise stated, the following terms shall have the meaning indicated.

[0023] It should be noted that, unless the context clearly indicates otherwise, the singular forms "a," "an," and "the" as used in this specification and the intended claims all include plural references. Thus, for example, reference to "a compound" includes a single compound as well as one or more identical or different compounds.

[0024] In some cases, the number of carbon atoms in a portion is indicated by the prefix "C xC y", where x is the minimum number of carbon atoms in the substituent and y is the maximum number. Thus, for example, "C 1-C 3 alkyl" means an alkyl substituent containing 1 to 3 carbon atoms.

[0025] Unless otherwise stated, the term "alkyl" as used herein refers to a saturated, straight-chain, or branched hydrocarbon radical having one, two, three, four, five, or six carbon atoms. The term "C1-C3 alkyl" refers to an alkyl group having one, two, or three carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, and the like. In some embodiments, the alkyl group is methyl. In some embodiments, the alkyl group is ethyl. In some embodiments, the alkyl group is propyl. In some embodiments, the alkyl group is isopropyl.

[0026] As used herein, the term "halo" means Cl (chlorine), Br (bromine), I (iodine), and F (fluorine). In some embodiments, the halo group is selected from the group consisting of Cl or F. In some embodiments, the halo group is Cl. In some embodiments, the halo group is F.

[0027] As used herein, the term "haloalkyl" means an alkyl group as defined herein, wherein one or more hydrogen atoms are replaced by halogen groups. As an example, the term "C1-C3 haloalkyl" refers to a haloalkyl group having one, two, or three carbon atoms. In some embodiments, one, two, or three hydrogen atoms may each be replaced by a fluorine group. Examples of haloalkyl groups are trifluoromethyl or CF3.

Implementation Method

[0029] In some embodiments, the compounds disclosed herein comprise two ligands linked via a linker. The first ligand is capable of binding Btk. The second ligand is capable of recruiting a ubiquitin ligase. In some embodiments, the ubiquitin ligase is an E3 ligase. In some embodiments, the E3 ubiquitin ligase is cerebellum (CRBN) and / or contains the cerebellum (CRBN). In some embodiments, the second ligand is capable of recruiting the CRBN. In some embodiments, the second ligand is capable of binding the CRBN. In some embodiments, the compounds described herein are capable of binding Btk and are capable of recruiting ubiquitin ligases containing the cerebellum. In some embodiments, the compounds disclosed herein recruit the E3 ligase to Btk and induce proximity-induced ubiquitination and degradation of Btk by the ubiquitin-proteasome system (UPS). In some embodiments, the compounds disclosed herein are capable of binding Btk and reducing the amount of Btk in cells through degradation.

[0030] In some embodiments, the compounds disclosed herein can be used to treat chronic lymphocytic leukemia (CLL). For example, the compounds described herein can be used to treat CLL by degrading Btk in patients via the ubiquitin protein hydrolysis pathway.

[0031] In some embodiments, the compounds disclosed herein degrade the Btk C481S protein (a Btk protein with a C481 mutation) in cell lines expressing Btk C481S.

[0032] In some embodiments, a method of treating a patient with CLL is provided, the method comprising administering the compound of formula (I) to the patient.

[0033] In some embodiments, a method for treating CLL is provided, the method comprising administering a therapeutically effective amount of the compound of formula (I) to a subject in need.

[0034] The phrase "pharmaceutically acceptable salt" refers to salts that, within the scope of reasonable medical judgment, are suitable for use in contact with human and lower animal tissues without abnormal toxicity, irritation, allergic reactions, etc., and that are commensurate with a reasonable benefit / risk ratio.

[0035] As used herein, the term "subject" refers to a human being. The terms "human," "patient," and "subject" are used interchangeably in this document.

[0036] As used herein, the terms “treat,” “treating,” and “treatment” refer to methods of alleviating or eliminating a disease and / or its accompanying symptoms.

[0037] The phrase "therapeutically effective amount" refers to an amount of a compound or a pharmaceutically acceptable salt thereof that, when administered to a specific subject or group of subjects, is sufficient to prevent or, to some extent, alleviate the development of one or more symptoms of the treated condition or symptom. In some embodiments, this amount effectively degrades Btk in the patient. General synthesis

[0038] The compounds disclosed herein can be better understood in conjunction with the following synthetic schemes and methods, which illustrate the means by which the compounds can be prepared. Representative procedures are shown in Schemes 1-5, but are not limited thereto. In Schemes 1-5, the variables R1, R2, R3, R4, and R5 can be as described herein. The final compounds of the present invention are named using the ACD / Name 2021.1.3 software program (file version N15E41, build 123232, July 7, 2021) and / or by using the Struct=Name naming algorithm as part of CHEMDRAW® Professional v.20.1.1.125. Intermediates are named using CHEMDRAW® Professional v.20.1.1.125.

[0039] The compounds of the present invention can be prepared according to the schemes described below. Scheme 1

[0040] Scheme 1 describes a method for preparing substituted 1-phenyldihydropyrimidine-2,4(1H,3H)-dione building blocks 1-6. As described herein, the substituent R5 on compound 1-1 may be a halogroup, hydrogen, or C1-C3 alkyl. The reaction of compound 1-1 with tributyldimethylchlorosilane in the presence of imidazole provides compound 1-2. The reaction of compound 1-2 with acrylic acid under heat forms compound 1-3. Compound 1-3 can be reacted with heated acetic acid to produce compound 1-4. Compound 1-4 can be treated with an aqueous solution of HCl to provide intermediate 1-5. Intermediate 1-5 can be treated with iodine in the presence of imidazole and then with Ph 3P to provide iodine compound 1-6, or it can be treated with CBr 4 and then with Ph 3P to provide bromine compound 1-6. Scheme 2

[0041] Scheme 2 describes a method for preparing the substituted building block 2-7. The reaction of the substituted benzaldehyde 2-1 with (S)-2-methylpropane-2-sulfinamide 2-2 provides compound 2-3, wherein R1, R2, and R3 are as described herein. Reaction of compound 2-3 with CH3MgBr under Grignard conditions yields intermediate 2-4. Intermediate 2-5 can be synthesized from intermediate 2-4 using bis(pinacolyl)diboron and a palladium catalyst. Amine 2-6 can be prepared from intermediate 2-5 by removing the tert-butylsulfinamide group from tert-butylamine using a strong acid or iodine. The heteroarylmethylamine building block 2-7 can be directly synthesized from amine 2-6 with methyl 3-(tert-butyl)-1,2,4-diazole-5-carboxylate. Scheme 3

[0042] As shown in Scheme 3, construct 3-4 can be prepared from compound 3-3 by hydrogenation or nitro reduction reaction, wherein R4a and R4b are as described herein. Intermediate compound 3-3 can be prepared from 2-nitro-5-halopyridine compound 3-1 and substituted piperidine compound 3-2 by S NAr reaction or Buchwald-Hartwing cross-coupling reaction. Scheme 4

[0043] The general synthesis of intermediate 4-5 is shown in Scheme 4, wherein R1, R2, R3, R4A, and R4b can be as described herein. Intermediate 4-2 can be prepared by the Suzuki reaction of building block 2-7 with 4,6-dichloropyrimidine 4-1. Intermediate 4-4 can be prepared by the Buchwald-Hartwing cross-coupling reaction of compound 4-2 with substituted 2-aminopyridine compound 3-4. The tertiary butoxycarbonyl protecting group can be cleaved with hydrochloric acid or trifluoroacetic acid to give building block 4-5. Scheme 5

[0044] The general synthesis of the target compound of formula 5-4 is shown in Scheme 5, wherein R1, R2, R3, R4A, R4b, and R5 may be as described herein. Iodine or bromine building blocks 1-6 may be reacted with intermediates 4-5 via alkylation using sodium iodide and a base such as N,N-diisopropylethylamine to provide target compound 5-4.

[0045] It is understood that the synthesis schemes and specific examples described in the Examples section are illustrative and should not be construed as limiting the scope of this disclosure as defined in the appended claims. All substitutions, modifications and equivalents of the synthesis methods and specific examples are included within the scope of the claims.

[0046] The following examples are for illustrative purposes and should not be construed as narrowing the scope of this disclosure.

[0047] The illustrative compounds of formula (I) include, but are not limited to, the compounds shown in Table 1 below and their pharmaceutically acceptable salts. Table 1. Illustrative Compounds Example Compound Structure 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 Example abbreviation definition NMR Nuclear magnetic resonance S Single peak br s Broad single peak D Duplet (or doublet) br d Broad double peak br dd Broad double double peak M Multiplets T Triple Peak br t Wide triple peak Td Triple Double Peak Q Four Peaks Qd Quadruple Double Peak Dd Double peak Dt Double triple peak Dq Double Quadruple Peak Ddd Double double double peak Dtd Double triple double peak Quin Five Peaks mL milliliters µL microliter L Lift G gram Mg mg Mol Mo Er Mmol Momo mM Momo M Mole concentration (mol / L) µM Micromer N Equivalent concentration (per liter) Ppm Million points Psi pounds per square inch HPLC High-performance liquid chromatography LC / MS or LCMS Liquid chromatography-mass spectrometry MS Mass spectrometry ESI Electrostatic ionization ATP Adenosine triphosphate BSA Bovine serum albumin EDTA EDTA-1,5-diaminetetraacetic acid DTT Dithiothreitol HEPES (4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid) IC 50 Half-maximum inhibition concentration DC 50 Half-maximum degradation concentration UV Ultraviolet rays TLC Thin-layer chromatography Mm millimeters Mm micrometer (micron) MHz million Hertz Nm nanometer ℃ Celsius m / z mass to charge ratio pH Hydrogen Index Pd / C Palladium on carbon

[0048] The final compound obtained by preparative HPLC may have been concentrated by lyophilization. Final samples are typically purified to ≥95% purity, as determined by HPLC and LCMS at 254 nm. 1H NMR spectra are recorded on a Bruker Avance™ III 400 MHz NMR spectrometer or a Bruker Ascend™ 500 MHz NMR spectrometer. For internal reference, chemical shifts (δ) are reported as parts per million (ppm) relative to residual undeuterated solvent. Example 1: 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine Example 1A (S, E)-N-(4-bromo-2-methylbenzyl)-2-methylpropane-2-sulfinamide

[0049] Triisopropyl borate (1.50 L, 6.53 mol), 4-bromo-2-methylbenzaldehyde (1.00 kg, 5.02 mol), and (S)-2-methylpropane-2-sulfinamide (669 g, 5.53 mol) were added to a 5 L reactor at ambient temperature. The mixture was stirred at 80 °C for 12 hours. The reaction was repeated twice on the same scale. Water (10.0 L) was added to the three batches of the combined reaction mixture, and the mixture was extracted with ethyl acetate (4.0 L × 2), washed with brine (5.0 L), dried with sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was milled in petroleum ether (5.0 L) for 2 hours and filtered. The filter cake was washed with petroleum ether and dried under reduced pressure to provide the title compound. ¹H NMR (400 MHz, CDCl₃): δppm 8.79 (s, ¹H), 7.77 (d, J=6.4 Hz, ¹H), 7.44 (s, ¹H), 7.43 (d, J=6.4 Hz, ¹H), 2.59 (s, 3H), 1.27 (s, 9H) ppm. Example 1B (S)-N-((R)-1-(4-bromo-2-methylphenyl)ethyl)-2-methylpropane-2-sulfinamide

[0050] Dichloromethane (20.0 L) and Example 1A (1.00 kg, 3.31 mol) were added to a 50 L reactor. The mixture was cooled at -20 °C. CH3MgBr (3.0 M contained in tetrahydrofuran, 1.65 L, 4.96 mol) was added dropwise at -20 °C. The mixture was then stirred at ambient temperature for 16 hours. The reaction mixture was cooled in an ice bath, and a saturated aqueous solution of NH4Cl (5.0 L) was added. The organic phase was separated, and the aqueous phase was extracted with dichloromethane (3.0 L × 2). The combined organic phases were washed with brine (5.0 L), dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was ground in petroleum ether. The crude material was isolated by filtration, washed with petroleum ether, and dried under reduced pressure. The same reaction was repeated five times to provide the title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d6): δppm 7.38–7.31 (m, 3H), 5.33 (d, J=5.2 Hz, 1H), 4.55 (m, 1H), 2.30 (s, 3H), 1.40 (d, J=5.2 Hz, 1H), 1.08 (s, 9H) ppm. MS (ESI) m / z 318.0, 320.0 (M+H)+. Example 1: C(S)-2-methyl-N-((R)-1-(2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethyl)propane-2-sulfinamide

[0051] At ambient temperature, Example 1B (600 g, 1.89 mol), dimethyl ether (3.6 L), bis(pinacolyl)diboron (600 g, 2.36 mol), potassium acetate (370 g, 3.77 mol), and Pd(dppf)Cl₂ ([1,1'-bis(diphenylphosphine)ferrocene]dichloropalladium(II), 23.0 g, 0.07 mol) were added to a 5 L reactor. The mixture was degassed with nitrogen and stirred at 85 °C for 8 hours under nitrogen. The same reaction was repeated four times on the same scale. Five batches of the combined reaction mixture were filtered through diatomaceous earth, and the filter cake was washed with ethyl acetate (0.5 L × 3). The filtrate was concentrated under reduced pressure, and the residue was subjected to rapid silica column chromatography using petroleum ether and ethyl acetate to provide the title compound. ¹H NMR (400 MHz, CDCl₃): δppm 7.58–7.38 (m, 3H), 5.31 (d, J=4.8 Hz, 1H), 4.60 (m, 1H), 2.31 (s, 3H), 1.40 (d, J=6.8 Hz, 1H), 1.27 (s, 12H), 1.08 (s, 9H) ppm. MS (ESI) m / z 366.2 (M+H)⁺. Example 1: D(R)-1-(2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethylamine

[0052] At 20°C, Example 1C (250 g, 680 mmol) and ethyl acetate (750 mL) were added to a 3 L reactor. The mixture was cooled to 0°C. At 0°C, hydrogen chloride / ethyl acetate (4 M, 750 mL) was added to the mixture. The reaction mixture was stirred at 20°C for 2 hours. The mixture was filtered, the solid product was ground in ethyl acetate, filtered, and dried under reduced pressure to give the crude title compound as a hydrochloride. ¹H NMR (400 MHz, dimethyl sulfoxide-d6): 8.62 (br s, 3 H), 7.52–7.60 (m, 3 H), 4.51–4.54 (br d, J = 5.2 Hz, 1 H), 2.35 (s, 3 H), 1.46 (d, J = 6.8 Hz, 3 H), 1.28 (s, 12 H) ppm. MS (ESI) m / z 262.1 (M+H) +. Example 1: E(R)-3-(tert-butyl)-N-(1-(2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethyl)-1,2,4-diazol-5-methylamine

[0053] At 20°C, triethylamine (64.0 mL, 459 mmol) was added to a solution of Example 1D (20 g, 77 mmol) and methyl 3-(tributyl)-1,2,4-diazole-5-carboxylate (31.0 g, 168 mmol) in ethanol (200 mL). The mixture was stirred at 65°C for 16 hours, followed by concentration under reduced pressure to remove ethanol. Water was added to the mixture, and the mixture was extracted three times with ethyl acetate. The combined organic layers were washed once with saturated ammonium chloride aqueous solution and once with brine, dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to provide crude material. The crude material was purified by silicone column chromatography, eluted with petroleum ether and ethyl acetate to provide the title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d⁶): δppm 9.83 (d, J=7.7 Hz, 1H), 7.53–7.43 (m, 3H), 5.28 (quin, J=7.1 Hz, 1H), 2.37 (s, 3H), 1.45 (d, J=7.0 Hz, 3H), 1.35 (s, 9H), 1.27 (s, 12H) ppm. MS (ESI) m / z 414.3 (M+H)⁺. Example 1F 4-(2-(tributyldimethylsilyl)oxy)ethyl)aniline

[0054] At ambient temperature, 2-(4-aminophenyl)-ethane-1-ol (176 g, 1.28 mol), N,N-dimethylformamide (1.8 L), imidazole (91.7 g, 1.35 mol), and tributyldimethylchlorosilane (222 g, 1.48 mol) were added to a 3 L reactor. The mixture was stirred at ambient temperature for 6 hours. Ethyl acetate (500 mL) and water (300 mL) were added to the mixture. The organic phase was separated, washed with brine (300 mL), dried over sodium sulfate, concentrated under reduced pressure, and the residue was subjected to rapid column chromatography (eluting with petroleum ether and ethyl acetate) to give the title compound. MS (ESI) m / z 252.2 (M+H)+. Example 1G 3-((4-(2-((tributyldimethylsilyl)oxy)ethyl)phenyl)amino)propionic acid

[0055] Example 1F (196 g, 0.78 mol) was added to a 3 L reactor. Toluene (1.4 L) and acrylic acid (67.4 g, 0.94 mol) were added to the mixture. The reaction mixture was stirred at 110 °C for 12 hours under nitrogen. The mixture was concentrated to dryness under reduced pressure to provide the crude title compound. MS (ESI) m / z 324.2 (M+H)+. Example 1H 4-(2,4-dioxytetrahydropyridinyl-1(2H)-yl)phenethyl acetate

[0056] Acetic acid (1.8 L) and urea (46.8 g, 0.78 mol) were added to crude Example 1G (252 g, 0.78 mol). The reaction mixture was stirred at 110 °C for 12 hours under nitrogen and cooled to ambient temperature. Water (600 mL) and ethyl acetate (600 mL) were added to the mixture. The organic phase was separated, and the aqueous phase was extracted with ethyl acetate (600 mL). The combined organic phases were washed with brine, dried over sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure to provide the crude title compound. MS (ESI) m / z 277.1 (M+H) +. Example 1I 1-(4-(2-hydroxyethyl)phenyl)dihydropyrimidine-2,4(1 H,3 H)-dione

[0057] A 6 N HCl aqueous solution (1.23 L, 7.39 mol) was added to crude Example 1H (176 g, 0.63 mol), and the mixture was stirred at ambient temperature for 6 hours. The reaction mixture was concentrated to dryness under reduced pressure, and the residue was milled with ethanol / ethyl acetate (250 mL / 250 mL) for 30 minutes. The material was isolated by filtration and dried under vacuum to provide the crude title compound. MS (ESI) m / z 235.1 (M+H) +. Example 1J 1-(4-(2-iodoethyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0058] To a 250 mL flask, add Example 1I (10.0 g, 0.04 mol) and dichloromethane (100 mL). In an ice bath, add iodine (16.3 g, 0.06 mol) in small portions, followed by Ph 3P (16.8 g, 0.06 mol), and then imidazole (4.36 g, 0.06 mol). Stir the reaction mixture at ambient temperature for 3 hours. Repeat the same reaction seven times in parallel on the same scale. Combine the eight batches of reaction mixture and pour into a mixture of water (800 mL) and ethyl acetate (800 mL). After vigorous stirring, separate the organic phase, filter, dry with sodium sulfate, and filter again. Concentrate the filtrate under reduced pressure to provide the title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d6): δppm 10.34 (s, ¹H), 7.32–7.24 (m, 4H), 3.75 (m, 2H), 3.49 (m, 2H), 3.13 (m, 2H), 2.72 (m, 2H) ppm. MS (ESI) m / z 345.0 (M+H)+. Example 1K(S)-3-methyl-4-(6-nitropyridin-3-yl)piperazine-1-carboxylic acid tributyl ester

[0059] At 20°C, tributyl (S)-3-methylpiperazine-1-carboxylate (37.9 g, 189 mmol), potassium phosphate (60.2 g, 284 mmol), and (R)-(+)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (5.89 g, 9.46 mmol) were added to a solution of 5-chloro-2-nitropyridine (30 g, 189 mmol) in toluene (600 mL). The reaction vessel was evacuated and backfilled with argon three times, and palladium(II) acetate (2.124 g, 9.46 mmol) was added in a single dose at 20°C. The vessel was then evacuated and backfilled with argon three times again. The reaction mixture was heated to 90°C and stirred at 90°C for 24 hours under an argon atmosphere. The reaction mixture was cooled, filtered through a diatomaceous earth mat, and washed with ethyl acetate. The filtrate was concentrated under reduced pressure, and the residues were purified by a silica rapid column (eluting from 0% to 50% with tetrahydrofuran in petroleum ether) to provide the title compound. 1H NMR (400 MHz, dimethyl sulfoxide-d 6): δ ppm 8.11-8.26 (m, 2 H), 7.42 (dd, J=9.38, 3.00 Hz, 1 H), 4.32 (br s, 1 H), 3.93 (br s, 1 H), 3.79 (br d, J=12.76 Hz, 2 H), 3.14-3.29 (m, 2 H), 3.06 (br s, 1 H), 1.42 (s, 9 H), 1.08 (d, J=6.63 Hz, 3 H) ppm. MS (ESI) m / z323.2 (M+H) +. Example 1: L(S)-4-(6-aminopyridin-3-yl)-3-methylpiperazine-1-carboxylic acid tributyl ester

[0060] Example 1K (70 g, 217 mmol) was added to a suspension of Pd / C (10 wt%, 46.2 g, 43.4 mmol) in tetrahydrofuran (700 mL), and the reaction mixture was stirred at ambient temperature under hydrogen (15 psi) for 24 hours. The same reaction was repeated on the same scale. The combined reaction mixture was filtered through a diatomaceous earth mat, and the filtrate was concentrated under reduced pressure to give a crude product. The material was ground with petroleum ether and ethyl acetate, the resulting solid was filtered, and the filter cake was dried under vacuum to provide the title compound. 1H NMR (400 MHz, CDCl 3): δppm 7.62 (d, J=2.63 Hz, 1 H), 7.20 (dd, J=8.76, 2.63 Hz, 1 H), 6.40 (d, J=8.76 Hz, 1 H), 5.56 (s, 2 H), 3.55 (br d, J=11.38 Hz, 1 H), 3.39 (br d, J=5.13 Hz, 2 H), 3.17 (br s, 2 H), 2.71 - 2.90 (m, 2 H), 1.41 (s, 9 H), 0.75 (d, J=6.25 Hz, 3 H) ppm. MS (ESI) m / z293.2 (M+H) +. Example 1: M(R)-3-(tert-butyl)-N-(1-(4-(6-chloropyrimidin-4-yl)-2-methylphenyl)ethyl)-1,2,4-diazol-5-methamide

[0061] At 20°C, a solution of potassium phosphate (10.27 g, 48.4 mmol) in water (100 mL) was added to a mixture of Example 1E (10 g, 24.19 mmol) and 4,6-dichloropyrimidine (7.21 g, 48.4 mmol) in tetrahydrofuran (400 mL). The mixture was degassed with a stream of nitrogen at ambient temperature, and [1,1'-bis(diphenylphosphine)ferrocene]dichloropalladium(II) (0.885 g, 1.210 mmol) was added to the mixture under N2. The reaction mixture was heated to 80°C and stirred for 2 hours. The mixture was cooled, poured into water, and extracted with ethyl acetate (three times). The combined organic layers were washed with brine, dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica column chromatography, eluted with tetrahydrofuran / petroleum ether to provide the crude title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d⁶): δppm 9.93 (d, J=7.8 Hz, ¹H), 9.07 (s, ¹H), 8.30 (s, ¹H), 8.09 (d, J=7.8 Hz, ¹H), 7.63 (d, J=8.5 Hz, ¹H), 5.33 (quin, J=7.2 Hz, ¹H), 2.48–2.47 (m, ¹H), 2.47 (s, 2H), 1.50 (d, J=7.0 Hz, 3H), 1.36 (s, 9H) ppm. MS (ESI) m / z 400.1 (M+H)⁺. Example 1: N(S)-4-(6-((6-(4-((R)-1-(3-(tributyl)-1,2,4-diazol-5-methamido)ethyl)-3-methylphenyl)pyrimidin-4-yl)amino)pyridin-3-yl)-3-methylpiperazine-1-carboxylic acid tributyl ester

[0062] At 20°C, Example 1M (34.2 g, 86 mmol), Cs₂CO₃ (55.7 g, 171 mmol), and (9,9-dimethyl-9H-dibenzopiperan-4,5-diyl)bis(diphenylphosphine) (4.95 g, 8.55 mmol) were added to a solution of Example 1L (25 g, 86 mmol) in 1,4-dimethylacetylene (500 mL). The reaction vessel was evacuated and backfilled with argon, and this process was repeated three times. Subsequently, ((tris(dibenzyl ethyl ketone)dipalladium(0)) (3.91 g, 4.28 mmol) was added under argon atmosphere. The reaction vessel was evacuated and backfilled with argon, and this process was repeated three times. The reaction mixture was heated to 100 °C and stirred at 100 °C for 16 hours. The reaction mixture was cooled, filtered through a diatomaceous earth mat, and washed with ethyl acetate. The filtrate was concentrated under reduced pressure, and the residue was purified by silica column chromatography, eluted with tetrahydrofuran / petroleum ether to provide the title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d6): δppm 10.02 (s, ¹H), 9.89 (d, J=7.88 Hz, ¹H), 8.71 (d, J=0.75 Hz, ¹H), 7.97 - 8.10 (m, ²H), 7.79 - 7.88 (m, 2 H), 7.71 (br d, J=8.50 Hz, 1 H), 7.61 (d, J=8.63 Hz, 1 H), 7.44 (dd, J=9.13, 2.88 Hz, 1 H), 5.33 (quin, J=7.13 Hz, 1 H), 3.73 - 3.92 (m, 2 H), 3.60 (br d, J=11.51 Hz, 1 H), 3.31 (br s, 1 H), 3.16 (br d, J=11.88 Hz, 2 H), 2.88 - 3.01 (m, 1 H), 2.47 (s, 3 H), 1.51 (d, J=6.88 Hz, 3 H), 1.42 (s, 9 H), 1.36 (s, 9 H), 0.88 (d, J=6.38 Hz, 3 H) ppm. MS (ESI) m / z 656.2 (M+H) +. Example 1 O 3-(tert-butyl)-N-((R)-1-(2-methyl-4-(6-((5-((S)-2-methylpiperazol-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)phenyl)ethyl)-1,2,4-diazol-5-methylamine

[0063] To a solution of Example 1N (32 g, 48.8 mmol) in dimethyl methacrylate (300 mL), the hydrochloride (4 N contained in dimethyl methacrylate, 300 mL) was added dropwise. The reaction mixture was stirred at ambient temperature for 2 hours. The reaction mixture was filtered to isolate the crude material and dried under vacuum. The crude material was diluted with water (300 mL), and a saturated aqueous solution of sodium bicarbonate was added to the mixture to adjust the pH to 8. The mixture was extracted twice with ethyl acetate, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to provide the title compound. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 10.00 (s, 1 H), 9.88 (d, J=7.78 Hz, 1 H), 8.70 (d, J=0.88 Hz, 1 H), 8.04 (br s, 1 H), 8.00 (d, J=2.89 Hz, 1 H), 7.79-7.89 (m, 2 H), 7.71 (br d, J=8.66 Hz, 1 H), 7.61 (d, J=8.66 Hz, 1 H) 7.43 (dd, J=9.16, 2.89 Hz, 1 H) 5.33 (quin, J=7.12 Hz, 1 H) 3.67-3.81 (m, 2 H), 3.67-3.81 (m, 1 H), 3.67-3.81 (m, 1 H), 3.08-3.17 (m, 1 H), 2.89-3.06 (m, 3 H), 2.74-2.89 (m, 2 H), 2.47 (s, 3 H), 1.51 (d, J=7.03 Hz, 3 H), 1.36 (s, 9 H), 0.96 (d, J=6.53 Hz, 3 H) ppm. MS (ESI) m / z556.2 (M+H) +. Example 1P 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine

[0064] At 20°C, Example 1J (18.36 g, 53.3 mmol), N,N-diisopropylethylamine (46.6 mL, 267 mmol), and sodium iodide (19.99 g, 133 mmol) were added to a solution of Example 1O (24.7 g, 44.5 mmol) in acetonitrile (500 mL). The reaction mixture was then stirred at 90°C for 16 hours. The reaction mixture was cooled, poured into ice water, and filtered. The filter cake was milled twice with ethyl acetate / tetrahydrofuran. The material was dried under reduced pressure to provide the title compound. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 10.33 (s, 1 H), 9.99 (s, 1 H), 9.89 (d, J=7.78 Hz, 1 H), 8.70 (s, 1 H), 7.96 - 8.09 (m, 2 H), 7.80 - 7.87 (m, 2 H), 7.70 (br d, J=7.65 Hz, 1 H), 7.61 (d, J=8.66 Hz, 1 H), 7.44 (dd, J=9.10, 2.82 Hz, 1 H), 7.19 - 7.32 (m, 4 H), 5.33 (br t, J=7.28 Hz, 1 H), 3.86 (br d, J=6.65 Hz, 1 H), 3.76 (t, J=6.65 Hz, 2 H), 3.21 (br d, J=11.92 Hz, 1 H), 2.99 (br t, J=9.35 Hz, 1 H), 2.84 (br d, J=10.79 Hz, 1 H), 2.77 (br t, J=7.47 Hz, 2 H), 2.69 (br t, J=6.65 Hz, 3 H), 2.56 (br s, 2 H), 2.47 (s, 3 H), 2.44 (br s, 1 H), 2.26 - 2.37 (m, 1 H), 1.51 (d, J=6.90 Hz, 3 H), 1.36 (s, 9 H), 0.99 (d, J=6.27 Hz, 3 H) ppm. MS (ESI) m / z772.2 (M+H) +.Example 2 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2R)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperidin-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-carboxamide Example 2A (R)-3-methyl-4-(6-nitropyridin-3-yl)piperidin-1-carboxylic acid tributyl ester.

[0065] The title compound was prepared according to the procedure used in Example 1K, with (R)-3-methylpiperazine-1-carboxylic acid tributyl ester substituted for (S)-3-methylpiperazine-1-carboxylic acid tributyl ester. ¹H NMR (400 MHz, dimethyl sulfoxide-d6): δ ppm 8.13–8.24 (m, 2H), 7.43 (dd, J=9.32, 3.06 Hz, 1H), 4.32 (br s, 1H), 3.85–4.03 (br s, 1H), 3.79 (br d, J=12.88 Hz, 2H), 2.97–3.30 (m, 3H), 1.43 (s, 9H), 1.08 (d, J=6.50 Hz, 3H) ppm. MS (ESI) m / z 323.2 (M+H) +. Example 2B (R)-4-(6-aminopyridin-3-yl)-3-methylpiperazine-1-carboxylic acid tributyl ester

[0066] The title compound was prepared according to the procedure used in Example 1L, with Example 2A replacing Example 1K. ¹H NMR (400 MHz, dimethyl sulfoxide-d⁶): δppm 7.62 (d, J=2.63 Hz, ¹H), 7.20 (dd, J=8.76, 2.75 Hz, ¹H), 6.40 (d, J=8.76 Hz, ¹H), 5.54 (s, ²H), 3.56 (br d, J=10.76 Hz, ¹H), 3.40 (br s, ²H), 3.17 (br s, ²H), 2.71–2.88 (m, ²H), 1.41 (s, ⁹H), 0.76 (d, J=6.13 Hz, ³H) ppm. MS (ESI) m / z 293.1 (M+H) +. Example 2C (R)-4-(6-((6-(4-((R)-1-(3-(tributyl)-1,2,4-diazol-5-methamido)ethyl)-3-methylphenyl)pyrimidin-4-yl)amino)pyridin-3-yl)-3-methylpiperazine-1-carboxylic acid tributyl ester

[0067] The title compound was prepared according to the procedure used in Example 1N, replacing Example 1L with Example 2B. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δ ppm 9.99 (s, 1 H), 9.87 (d, J=7.78 Hz, 1 H), 8.70 (d, J=1.00 Hz, 1 H), 7.98 - 8.07 (m, 2 H), 7.79 - 7.88 (m, 2 H), 7.71 (br d, J=9.16 Hz, 1 H), 7.60 (d, J=8.66 Hz, 1 H), 7.45 (dd, J=9.10, 2.82 Hz, 1 H), 5.33 (quin, J=7.25 Hz, 1 H), 3.75 - 3.88 (m, 2 H), 3.56 - 3.65 (m, 1 H), 3.11 - 3.22 (m, 2 H), 2.90 (s, 1 H), 2.52 (br s, 1 H), 2.47 (s, 3 H), 1.51 (d, J=7.03 Hz, 3 H), 1.42 (s, 9 H), 1.36 (s, 9 H), 0.89 (d, J=6.40 Hz, 3 H) ppm. MS (ESI) m / z 656.3 (M+H) +. Example 2D 3-(tributyl)-N-((R)-1-(2-methyl-4-(6-((5-(((R)-2-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)phenyl)ethyl)-1,2,4-diazol-5-methylamine

[0068] The title compound was prepared according to the procedure used in Example 10, with Example 2C replacing Example 1N. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δ ppm 9.97 (s, 1 H), 9.87 (d, J=7.78 Hz, 1 H), 8.70 (d, J=1.00 Hz, 1 H), 8.03 (br s, 1 H), 7.98 (d, J=2.76 Hz, 1 H), 7.80 - 7.87 (m, 2 H), 7.69 (br d, J=8.66 Hz, 1 H), 7.60 (d, J=8.66 Hz, 1 H), 7.41 (dd, J=9.16, 2.89 Hz, 1 H), 5.26 - 5.40 (m, 1 H), 3.65 - 3.77 (m, 1H), 3.01 - 3.15 (m, 1 H), 2.90 - 2.98 (m, 2 H), 2.84 - 2.90 (m, 1 H), 2.69 - 2.79 (m, 2 H), 2.52 (br d, J=1.88 Hz, 1 H), 2.47 (s, 3 H), 1.51 (d, J=6.90 Hz, 3 H), 1.36 (s, 9 H), 0.96 (d, J=6.53 Hz, 3 H) ppm. MS (ESI) m / z556.4 (M+H) +. Example 2E 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2R)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine

[0069] The title compound was prepared according to the procedure used in Example 1P, replacing Example 1O with Example 2D. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 10.31 (br s, 1 H), 9.97 (s, 1 H), 9.87 (br d, J=6.02 Hz, 1 H), 8.70 (d, J=0.88 Hz, 1 H), 7.94 - 8.10 (m, 2 H), 7.78 - 7.89 (m, 2 H), 7.70 (br d, J=8.03 Hz, 1 H), 7.60 (d, J=8.66 Hz, 1 H), 7.43 (dd, J=9.16, 2.89 Hz, 1 H), 7.18 - 7.32 (m, 4 H), 5.24 - 5.42 (m, 1 H), 3.81 - 3.90 (m, 1 H), 3.76 (t, J=6.65 Hz, 2 H), 3.16 - 3.26 (m, 1 H), 2.93 - 3.05 (m, 1 H), 2.84 (br d, J=10.67 Hz, 1 H), 2.73 - 2.81 (m, 2 H), 2.67 - 2.73 (m, 3 H), 2.53 - 2.62 (m, 2 H), 2.47 (s, 3 H), 2.44 (br s, 1 H), 2.27 - 2.36 (m, 1 H), 1.51 (d, J=7.03 Hz, 3 H), 1.36 (s, 9 H), 0.99 (d, J=6.40 Hz, 3 H) ppm. MS (ESI) m / z 772.2 (M+H) +. Example 3 3-Tributyl-N-{(1R)-1-[4-(6-{[5-(4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}piperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine Example 3A 4-(6-nitropyridin-3-yl)piperazine-1-carboxylic acid tributyl ester

[0070] The title compound was prepared according to the procedure used in Example 1K, with (S)-3-methylpiperazine-1-carboxylate tributyl ester substituted for (S)-3-methylpiperazine-1-carboxylate tributyl ester. MS (ESI) m / z 309.2 (M+H) +. Example 3B 4-(6-aminopyridin-3-yl)piperazine-1-carboxylate tributyl ester

[0071] The title compound was prepared according to the procedure used in Example 1L, replacing Example 1K with Example 3A. MS (ESI) m / z 279.3 (M+H)+. Example 3C (R)-4-(6-((6-(4-(1-(3-(tributyl)-1,2,4-diazol-5-methamino)ethyl)-3-methylphenyl)pyrimidin-4-yl)amino)pyridin-3-yl)piperazine-1-carboxylic acid tributyl ester

[0072] The title compound was prepared according to the procedure used in Example 1N, replacing Example 1L with Example 3B. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δ ppm 10.03 (s, 1H), 9.90 (d, J= 7.8 Hz, 1H), 8.71 (s, 1H), 8.06 (br d, J= 2.9 Hz, 2H), 7.85 - 7.80 (m, 2H), 7.71 (br d, J= 8.0 Hz, 1H), 7.60 (d, J= 8.6 Hz, 1H), 7.47 (dd, J= 2.9, 9.1 Hz, 1H), 5.38 - 5.28 (m, 1H), 3.46 (br d, J= 4.8 Hz, 4H), 3.12 - 3.02 (m, 4H), 2.47 (s, 3H), 1.51 (d, J = 6.9 Hz, 3H), 1.42 (s, 9H), 1.37 - 1.34 (m, 9H) ppm. MS (ESI) m / z 642.4 (M+H)+. Example 3D(R)-3-(tert-butyl)-N-(1-(2-methyl-4-(6-((5-(piperidin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)phenyl)ethyl)-1,2,4-diazol-5-methylamine

[0073] The title compound was prepared according to the procedure used in Example 10, with Example 3C replacing Example 1N. ¹H NMR (400 MHz, dimethyl sulfoxide-d⁶): δppm 10.01 (s, ¹H), 9.89 (d, J=7.8 Hz, ¹H), 8.71 (s, ¹H), 8.14–7.95 (m, 2H), 7.89–7.77 (m, 2H), 7.71 (br d, J=8.4 Hz, 1H), 7.60 (d, J=8.8 Hz, 1H), 7.46 (dd, J=2.9, 9.1 Hz, 1H), 5.33 (quin, J=7.1 Hz, 1H), 3.19–3.11 (m, 4H), 3.05–2.96 (m, 4H), 2.47 (s, 3H), 1.51 (d, J=7.0 Hz, 3H), 1.36 (s, 9H) ppm. MS (ESI) m / z 542.3 (M+H) +. Example 3E 3-Tributyl-N-{(1R)-1-[4-(6-{[5-(4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}piperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine

[0074] The title compound was prepared according to the procedure used in Example 1P, replacing Example 1O with Example 3D. ¹H NMR (500 MHz, dimethyl sulfoxide-d⁶): δppm 10.34 (s, ¹H), 9.99 (s, ¹H), 9.88 (d, J = 7.8 Hz, ¹H), 8.70 (s, ¹H), 8.08–8.00 (m, 2H), 7.85–7.80 (m, 2H), 7.69 (brs, ¹H), 7.63–7.58 (m, ¹H), 7.47 (dd, J = 9.1, 3.0 Hz, 1H), 7.30–7.21 (m, J = 8.3 Hz, 4H), 5.33 (p, J = 7.1 Hz, 1H), 3.76 (t, J = 6.7 Hz, 2H). 3.14 (brs, 4H), 2.79 (brs, 2H), 2.70 (t, J= 6.7 Hz, 2H), 2.61 (brs, 5H), 2.47 (s, 3H), 1.51 (d, J= 7.0 Hz, 3H), 1.36 (s, 9H) ppm. MS (ESI) m / z758.3 (M+H) +. Example 4 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)-3-fluorophenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine Example 4A (E)-2-(3-fluoro-4-nitrophenyl)-N,N-dimethylethylene-1-amine

[0075] A mixture of 2-fluoro-4-methyl-1-nitrobenzene (1.50 kg, 9.63 mol) and N,N-dimethylformamide-dimethyl acetal (1.72 kg, 14.5 mol, 1.92 L) in N,N-dimethylformamide (2.4 L) was degassed and purged three times with nitrogen. The mixture was stirred under nitrogen at 120 °C (internal reading) for 2 hours. The reaction mixture was concentrated under reduced pressure. The crude material was milled with petroleum ether / methyl tributyl ether (1:1, 10.0 L) and filtered. The filter cake was washed with petroleum ether / methyl tributyl ether (1:1, 2.0 L) and dried under vacuum to provide the title compound. ¹H NMR (400 MHz, CDCl₃): δppm 7.92 (t, J = 8.4 Hz, ¹H), 7.03 (d, J = 13.6 Hz, ¹H), 6.89–6.84 (m, 2H), 5.05 (d, J = 13.2 Hz, ¹H), 2.95 (s, 6H) ppm. Example 4B: 2-(3-fluoro-4-nitrophenyl)ethane-1-ol

[0076] At ambient temperature, a small amount of Example 4A (1.00 kg, 4.76 mol) was added to an aqueous hydrochloric acid solution (4 M, 7.50 L). The mixture was stirred at ambient temperature for 12 hours and extracted with 2-methyltetrahydrofuran (3.0 L × 7). The combined organic phases were washed with brine (5.0 L) and dried over sodium sulfate. After filtration, the filtrate was concentrated to a volume of 10 L under reduced pressure and cooled in an ice bath. A small amount of sodium borate (214 g, 5.68 mol) was added to the mixture in an ice bath, and the reaction mixture was stirred at ambient temperature for 16 hours. The mixture was cooled in an ice bath and quenched with an aqueous hydrochloric acid solution (5.0 L). The organic phase was separated, washed with brine (3.0 L), dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to provide the crude title compound. Example 4C: Tributyl(3-fluoro-4-nitrophenylethoxy)dimethylsilane

[0077] At ambient temperature, tributyldimethylchlorosilane (804 g, 5.34 mol, 654 mL) was added aliquots to a mixture of crude Example 4B (660 g, 3.56 mol) and imidazole (364 g, 5.34 mol) in tetrahydrofuran (4.4 L). The mixture was stirred at ambient temperature for 12 hours, quenched by the addition of water (3.0 L), and diluted with ethyl acetate (3.0 L). The organic layer was separated, washed with brine (5.0 L), dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The residue was subjected to silica rapid column chromatography (petroleum ether and ethyl acetate) to provide the title compound. ¹H NMR (400 MHz, CDCl₃): δppm 7.92–8.05 (m, ¹H), 7.11–7.20 (m, 2H), 3.79–3.92 (m, 2H), 2.80–2.93 (m, 2H), 0.80 (s, 9H), -0.09 (s, 6H) ppm. Example 4D 4-(2-((tributyldimethylsilyl)oxy)ethyl)-2-fluoroaniline

[0078] At 25°C, Example 4C (5.5 g, 18.37 mmol) was added to a suspension of Pd / C (2.346 g, 2.204 mmol) in tetrahydrofuran (100 mL). The resulting mixture was stirred at 25°C for 12 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure at 40°C to give the title compound. MS (ESI) m / z 270.1 (M+H)+. Example 4E 3-((4-(2-((tributyldimethylsilyl)oxy)ethyl)-2-fluorophenyl)amino)propionic acid

[0079] Acrylic acid (0.802 g, 11.13 mmol) was added to a solution of Example 4D (2.5 g, 9.28 mmol) in toluene (25 mL). The reaction mixture was heated to 120 °C and stirred for 16 hours. The reaction mixture was cooled to 20 °C and concentrated under reduced pressure to give the crude title compound. MS (ESI) m / z 342.1 (M+H)+. Example 4F 4-(2,4-dioxytetrahydropyridinyl-1(2H)-yl)-3-fluorophenylethyl acetate

[0080] Urea (1.583 g, 26.4 mmol) was added to a solution of Example 4E (3 g, 8.78 mmol) in acetic acid (30 mL). The reaction mixture was heated to 120 °C and stirred for 16 hours. The reaction mixture was cooled to 20 °C and concentrated under reduced pressure. The crude material was purified by preparative HPLC (Phenomenex Luna® C18 column, 50*250 mm, 10 µm, using 0.1% formic acid / water and acetonitrile as the mobile phase) to provide the title compound. MS (ESI) m / z 295.1 (M+H)+. Example 4G 1-(2-fluoro-4-(2-hydroxyethyl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0081] At 20°C, concentrated hydrochloric acid (37%, 1 mL) was added to a solution of Example 4F (1.1 g, 3.74 mmol) in water (1 mL). The reaction mixture was stirred at 20°C for 12 hours. The reaction mixture was concentrated under reduced pressure to provide the title compound. MS (ESI) m / z 253.1 (M+H) +. Example 4H 1-(4-(2-bromoethyl)-2-fluorophenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0082] At 0°C, CBr 4 (0.789 g, 2.379 mmol) was added to a solution of Example 4G (0.3 g, 1.189 mmol) in dichloromethane (20 mL), followed by Ph 3P (0.624 g, 2.379 mmol). The reaction mixture was stirred at 20°C for 12 hours. The reaction mixture was concentrated under reduced pressure at 40°C. The residue was purified by silica rapid column chromatography (ethyl acetate / petroleum ether, 0-100%) to give the title compound. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 10.47 (s, 1H), 7.37 (t, J= 8.0 Hz, 1H), 7.27 (br d, J= 11.2 Hz, 1H), 7.16 (br d, J= 7.6 Hz, 1H), 3.77 (t, J= 7.2 Hz, 2H), 3.70 (t, J= 6.8 Hz, 2H), 3.16 (t, J= 7.2 Hz, 2H), 2.71 (t, J= 6.8 Hz, 2H) ppm. MS (ESI) m / z315.0, 317.0 (M+H) +. Example 4I 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)-3-fluorophenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine

[0083] Example 1O (80 mg, 0.14 mmol) was added to a solution of N,N-dimethylformamide (1 mL) containing Example 4H (54 mg, 0.17 mmol), sodium iodide (65 mg, 0.43 mmol), and N,N-diisopropylethylamine (56 mg, 0.43 mmol). The mixture was stirred at 80 °C for 16 hours. The mixture was cooled to ambient temperature and filtered. The filtrate was directly subjected to reversed-phase preparative HPLC to provide the title compound (Gilson GX-281 system, Waters™ Xbridge C18 column, 30 × 100 mm × 10 µm, 10 mM ammonium bicarbonate / water and acetonitrile as mobile phase). 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 10.46 (s, 1 H), 9.99 (s, 1 H), 9.89 (d, J=7.89 Hz, 1 H), 8.70 (d, J=0.66 Hz, 1 H), 7.96 - 8.08 (m, 2 H), 7.80 - 7.86 (m, 2 H), 7.66 - 7.75 (m, 1 H), 7.60 (d, J=8.77 Hz, 1 H), 7.44 (dd, J=9.21, 2.85 Hz, 1 H), 7.33 (t, J=8.11 Hz, 1 H), 7.24 (dd, J=11.62, 1.53 Hz, 1 H), 7.15 (dd, J=8.11, 1.53 Hz, 1 H), 5.33 (t, J=7.34 Hz, 1 H), 3.82 - 3.91 (m, 1 H), 3.69 (t, J=6.69 Hz, 2 H), 3.21 (br d, J=11.84 Hz, 1 H), 2.98 (br t, J=9.32 Hz, 1 H), 2.77 - 2.83 (m, 2 H), 2.69 - 2.72 (m, 2 H), 2.53 - 2.64 (m, 2 H), 2.52 (d, J=1.75 Hz, 2 H), 2.47 (s, 3 H), 2.41 - 2.46 (m, 1 H), 2.26 - 2.32 (m, 1 H), 1.51 (d, J=7.02 Hz, 3 H), 1.36 (s, 9 H), 0.98 (d, J=6.36 Hz, 3 H) ppm. LC / MS (ESI+) m / z790.3 (M+H) +.Example 5 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2R)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)-3-fluorophenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine.

[0084] Example 2D (100 mg, 0.18 mmol) was added to a solution of N,N-dimethylformamide (2 mL) with Example 4H (68 mg, 0.22 mmol), sodium iodide (81 mg, 0.54 mmol), and N,N-diisopropylethylamine (70 mg, 0.54 mmol). The mixture was stirred at 80 °C for 16 hours. The mixture was cooled to ambient temperature and filtered. The filtrate was directly subjected to reversed-phase preparative HPLC to isolate the title compound (Gilson GX-281 system, Phenomenex Luna® C18 column, 30 × 75 mm × 3 µm, 0.1% formic acid / water and acetonitrile as mobile phase). 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 10.45 (s, 1 H), 9.99 (s, 1 H), 9.88 (d, J=7.82 Hz, 1 H), 8.70 (s, 1 H), 7.98 - 8.07 (m, 2 H), 7.79 - 7.87 (m, 2 H), 7.66 - 7.74 (m, 1 H), 7.60 (d, J=8.68 Hz, 1 H), 7.40 - 7.48 (m, 1 H), 7.33 (t, J=8.19 Hz, 1 H), 7.21 - 7.28 (m, 1 H), 7.15 (d, J=8.31 ​​Hz, 1 H), 5.24 - 5.44 (m, 1 H), 3.76 - 3.97 (m, 1 H), 3.69 (t, J=6.66 Hz, 2 H), 3.13 - 3.26 (m, 2 H), 2.94 - 3.06 (m, 1 H), 2.77 - 2.92 (m, 3 H), 2.71 (t, J=6.66 Hz, 3 H), 2.56 - 2.65 (m, 1 H), 2.47 (s, 3 H), 2.33 - 2.46 (m, 2 H), 1.51 (d, J=6.85 Hz, 3 H), 1.36 (s, 9 H), 0.98 (d, J=6.24 Hz, 3 H) ppm. LC / MS (ESI+): m / z790.2 (M+H)+.Example 6 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-ethylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-carboxamide Example 6A (S)-3-ethyl-4-(6-nitropyridin-3-yl)piperazine-1-carboxylic acid tributyl ester.

[0085] 5-Fluoro-2-nitropyridine (1.00 g, 7.04 mmol) and tributyl (S)-3-ethylpiperazine-1-carboxylic acid (1.96 g, 9.15 mmol) were dissolved in N,N-dimethylformamide, and potassium carbonate (2.92 g, 21.11 mmol) was added. The reaction mixture was stirred at 70 °C for 15 hours and concentrated under reduced pressure to remove most of the solvent. The residue was diluted with dichloromethane, washed with water, dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give the crude title compound. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δ ppm 8.21 (d, J=2.88 Hz, 1 H), 8.15 (d, J=9.26 Hz, 1 H), 7.42 (dd, J=9.32, 2.94 Hz, 1 H), 3.89 - 4.17 (m, 3 H), 3.82 (br d, J=12.88 Hz, 1 H), 2.90 - 3.27 (m, 3 H), 1.47 - 1.69 (m, 2 H), 1.42 (s, 9 H), 0.90 (br s, 3 H) ppm. LC / MS (ESI+): m / z337.0 (M+H)+. Example 6B (S)-4-(6-aminopyridin-3-yl)-3-ethylpiperazine-1-carboxylic acid tributyl ester

[0086] Example 6A (2.0 g, 5.95 mmol) was dissolved in methanol (200 mL). Pd / C (10 wt%, 0.32 g, 3.0 mmol) was added to the mixture. The reaction mixture was hydrogenated using a hydrogen balloon at ambient temperature for 2 days. The mixture was filtered through diatomaceous earth, and the solid cake was washed with methanol and ethyl acetate. The filtrate was concentrated under reduced pressure to give the crude title compound. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δ ppm 7.59 (br d, J=2.01 Hz, 1 H), 7.12 - 7.21 (m, 1 H), 6.40 (d, J=8.78 Hz, 1 H), 5.46 (br s, 2 H), 3.47 - 3.76 (m, 2 H), 2.98 - 3.23 (m, 2 H), 2.85 (br d, J=4.64 Hz, 2 H), 2.52 (br d, J=1.88 Hz, 1 H), 1.41 (s, 9 H), 1.35 (s, 1 H), 1.25 (br s, 1 H), 0.77 (br t, J=7.22 Hz, 3 H) ppm. LC / MS (ESI +): m / z 307.2 (M+H) +. Example: 6C(S)-4-(6-((6-(4-((R)-1-(3-(tributyl)-1,2,4-diazol-5-methamino)ethyl)-3-methylphenyl)pyrimidin-4-yl)amino)pyridin-3-yl)-3-ethylpiperazine-1-carboxylic acid tributyl ester.

[0087] To a solution of Example 1E (300 mg, 0.75 mmol) in 1,4-dimethylamine (6 mL), Example 6B (253 mg, 0.83 mmol), Pd₂(dba)₃ (CAS 52409-22-0, 34 mg, 0.038 mmol), XantPhos (CAS 161265-03-8, 43 mg, 0.075 mmol) and cesium carbonate (489 mg, 1.50 mmol) were added. The mixture was degassed using an argon stream and stirred at 100 °C for 16 hours. The reaction mixture was cooled, diluted with ethyl acetate, and filtered through diatomaceous earth. The filtrate was concentrated under reduced pressure and purified by silica chromatography using ethyl acetate and petroleum ether to give the title compound. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δ ppm 9.98 (s, 1 H), 9.89 (d, J=7.78 Hz, 1 H), 8.70 (s, 1 H), 7.96 - 8.07 (m, 2 H), 7.80 - 7.86 (m, 2 H), 7.66 - 7.74 (m, 1 H), 7.60 (d, J=8.66 Hz, 1 H), 7.38 - 7.48 (m, 1 H), 5.33 (quin, J=7.09 Hz, 1 H), 3.90 (br d, J=11.04 Hz, 2 H), 3.60 (br dd, J=6.53, 3.01 Hz, 1 H), 3.12 - 3.29 (m, 2 H), 2.89 - 3.05 (m, 2 H), 2.47 (s, 3 H), 1.61 (s, 1 H), 1.51 (br d, J=6.90 Hz, 3 H), 1.42 (s, 9 H), 1.36 (s, 9 H), 1.21 - 1.31 (m, 1 H), 0.86 (br s, 3 H) ppm. LC / MS (ESI+): m / z670.4 (M+H) +. Example 6D 3-(tert-butyl)-N-((R)-1-(4-(6-((5-((S)-2-ethylpiperazol-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)-2-methylphenyl)ethyl)-1,2,4-diazol-5-methylamine

[0088] At ambient temperature, hydrochloride (4N contained in diane, 3 mL) was added dropwise to a solution of Example 6C (150 mg, 0.22 mmol) in 1,4-diane (3 mL). The mixture was stirred for 2 hours, concentrated under reduced pressure, diluted with water (5 mL), and treated with a saturated aqueous sodium carbonate solution to adjust the pH to 8. The mixture was extracted with ethyl acetate (15 mL, three times), and the organic phase was dried over sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give the crude title compound. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δ ppm 9.94 (s, 1 H), 9.89 (d, J=7.78 Hz, 1 H), 8.69 (s, 1 H), 7.92 - 8.05 (m, 2 H), 7.80 - 7.87 (m, 2 H), 7.63 - 7.72 (m, 1 H), 7.60 (d, J=8.66 Hz, 1 H), 7.37 (dd, J=9.03, 2.89 Hz, 1 H), 5.33 (quin, J=7.09 Hz, 1 H), 3.45 (br dd, J=6.71, 2.45 Hz, 1 H), 3.09 - 3.17 (m, 1 H), 2.80 - 2.94 (m, 4 H), 2.66 - 2.74 (m, 1 H), 2.47 (s, 3 H), 1.75 (ddd, J=13.61, 9.47, 7.40 Hz, 1 H), 1.51 (d, J=6.90 Hz, 3 H), 1.36 (s, 9 H), 1.22 - 1.30 (m, 1 H), 0.79 (t, J=7.47 Hz, 3 H) ppm. LC / MS (ESI+): m / z570.4 (M+H) +. Example 6E 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-ethylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine

[0089] Example 6D (480 mg, 0.79 mmol), Example 1J (327 mg, 0.95 mmol), N,N-diisopropylethylamine (277 µL, 1.58 mmol), and potassium iodide (657 mg, 3.96 mmol) were mixed in N,N-dimethylformamide (8 mL). The mixture was stirred at 80 °C for 4 hours. The mixture was cooled, quenched with a few drops of formic acid, and directly subjected to reversed-phase preparative HPLC to isolate the title compound (Phenomenex Gemini® AXA C18 column, 30 × 250 mm × 5 µm, 0.1% formic acid / water and acetonitrile as mobile phase). 1H NMR (500 MHz, dimethyl sulfoxide - d 6): δppm 9.89 (s, 1H), 9.53 (s, 1H), 9.38 (d, J = 7.6 Hz, 1H), 8.64 (d, J = 1.1 Hz, 1H), 7.94 (dd, J = 11.8, 2.1 Hz, 2H), 7.84 – 7.78 (m, 2H), 7.64 (d, J = 9.0 Hz, 1H), 7.58 (d, J = 8.8 Hz, 1H), 7.33 (dd, J = 9.1, 3.1 Hz, 1H), 7.28 – 7.18 (m, 4H), 5.34 (p, J = 7.1 Hz, 1H), 3.73 (t, J = 6.7 Hz, 2H), 3.53 (s, 1H), 3.24 (d, J = 12.2 Hz, 1H), 3.10 – 3.02 (m, 1H), 2.79 (ddt, J = 14.1, 7.8, 3.6 Hz, 4H), 2.68 (t, J = 6.7 Hz, 2H), 2.60 (s, 2H), 2.45 (s, 3H), 2.41 – 2.27 (m, 3H), 1.73 – 1.59 (m, 1H), 1.52 (d, J = 7.0 Hz, 3H), 1.35 (s, 9H), 0.79 (t, J = 7.4 Hz, 3H) ppm. LC / MS (ESI+): m / z786.7 (M+H) +.Example 7 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)-3-fluorophenyl]ethyl}-2-ethylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine.

[0090] Example 6D (120 mg, 0.21 mmol), Example 4H (100 mg, 0.32 mmol), N,N-diisopropylethylamine (184 µL, 1.05 mmol), and sodium iodide (95 mg, 0.63 mmol) were mixed in acetonitrile (2.4 mL). The mixture was stirred at 80 °C for 16 hours. The mixture was cooled, quenched with a few drops of formic acid, and directly subjected to reversed-phase preparative HPLC to isolate the title compound (Phenomenex Luna® C18 column, 30 × 75 mm × 3 µm, 0.1% formic acid / water and acetonitrile as mobile phase). 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 10.46 (s, 1 H), 9.96 (s, 1 H), 9.89 (d, J=7.75 Hz, 1 H), 8.69 (s, 1 H), 7.94 - 8.05 (m, 2 H), 7.80 - 7.86 (m, 2 H), 7.64 - 7.73 (m, 1 H), 7.60 (d, J=8.63 Hz, 1 H), 7.37 - 7.44 (m, 1 H), 7.33 (t, J=8.19 Hz, 1 H), 7.21 - 7.28 (m, 1 H), 7.08 - 7.17 (m, 1 H), 5.25 - 5.41 (m, 1 H), 3.68 (t, J=6.63 Hz, 2 H), 3.57 - 3.65 (m, 1 H), 3.00 (br t, J=10.57 Hz, 1 H), 2.86 - 2.95 (m, 2 H), 2.81 (br d, J=6.38 Hz, 2 H), 2.71 (br t, J=6.63 Hz, 2 H), 2.60 - 2.65 (m, 1 H), 2.47 (br s, 5 H), 2.21 - 2.31 (m, 2 H), 1.62 - 1.76 (m, 1 H), 1.51 (d, J=7.00 Hz, 3 H), 1.36 (s, 9 H), 1.22 - 1.31 (m, 1 H), 0.81 (br t, J=7.38 Hz, 3 H) ppm. LC / MS (ESI+): m / z804.3 (M+H) +.Example 8 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2R)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-ethylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-carboxamide Example 8A (R)-3-ethyl-4-(6-nitropyridin-3-yl)piperazine-1-carboxylic acid tributyl ester.

[0091] The title compound was prepared according to the procedure used in Example 6A, with (R)-3-ethylpiperazine-1-carboxylic acid tributyl ester substituted for (S)-3-ethylpiperazine-1-carboxylic acid tributyl ester. LC / MS (ESI +): m / z 337.0 (M+H) +. Example 8B: (R)-4-(6-aminopyridin-3-yl)-3-ethylpiperazine-1-carboxylic acid tributyl ester

[0092] The title compound was prepared according to the procedure used in Example 6B, replacing Example 6A with Example 8A. LC / MS (ESI+): m / z 307.2 (M+H)+. Example 8C (R)-4-(6-((6-(4-((R)-1-(3-(tributyl)-1,2,4-diazol-5-methamino)ethyl)-3-methylphenyl)pyrimidin-4-yl)amino)pyridin-3-yl)-3-ethylpiperazine-1-carboxylic acid tributyl ester

[0093] The title compound was prepared according to the procedure used in Example 6C, replacing Example 6B with Example 8B. LC / MS (ESI+): m / z 670.1 (M+H)+. Example 8D 3-(tributyl)-N-((R)-1-(4-(6-((5-(((R)-2-ethylpiperazol-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine

[0094] The title compound was prepared according to the procedure used in Example 6D, replacing Example 6C with Example 8C. LC / MS (ESI+): m / z 570.7 (M+H)+. Example 8E 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2R)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-ethylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine

[0095] The title compound was prepared according to the procedure used in Example 6E, replacing Example 6D with Example 8D. ¹H NMR (400 MHz, dimethyl sulfoxide-d⁶): δ 9.95 (s, ¹H), 9.79 (s, ¹H), 9.40 (d, J = 7.5 Hz, ¹H), 8.70 (s, ¹H), 8.09 (s, ¹H), 7.99 (s, ¹H), 7.82 (d, J = 7.8 Hz, 2H), 7.74 (d, J = 8.7 Hz, 1H), 7.59 (d, J = 7.9 Hz, 1H), 7.51 (s, ¹H), 7.33–7.28 (m, 4H), 5.34 (p, J = 7.1 Hz, 1H), 3.75 (t, J = 6.7 Hz, 2H). 3.40-3.32 (m, 4H), 3.28 (s, 2H), 3.20 (s, 1H), 3.10-3.03 (m, 1H), 3.00-2.90 (m, 3H), 2.69 (t, J = 6.7 Hz, 2H), 2.46-2.45 (m, 5H), 1.52 (d, J = 6.9 Hz, 3H), 1.35 (s, 9H), 0.83 (t, J = 7.1 Hz, 3H) ppm. LC / MS (ESI+): m / z786.9 (M+H) +. Example 9: 3-Tributyl-N-{(1R)-1-[4-(6-{[5-(4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2,2-dimethylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-carboxamide Example 9A: 3,3-Dimethyl-4-(6-nitropyridin-3-yl)piperazine-1-carboxylic acid tributyl ester

[0096] 5-Bromo-2-nitropyridine (5.68 g, 28.0 mmol) and tributyl 3,3-dimethylpiperazine-1-carboxylate (3.0 g, 14.00 mmol) were dissolved in dimethyl ether (150 mL). Cesium carbonate (9.12 g, 28.0 mmol), BINAP ((R / S)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl, 0.872 g, 1.40 mmol), and tris(dibenzylacetone)dipalladium(O) (1.282 g, 1.40 mmol) were added to the mixture. The reaction mixture was stirred at 120 °C for 15 hours under a nitrogen atmosphere. The mixture was cooled and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica chromatography (ethyl acetate and hexane) to give the title compound. LC / MS (ESI+) m / z 336.9 (M+H) +. Example 9B: Tributyl 4-(6-aminopyridin-3-yl)-3,3-dimethylpiperazine-1-carboxylic acid.

[0097] Example 9A (1.1 g, 3.27 mmol) was dissolved in methanol (100 mL). Pd / C (10 wt%, 0.17 g, 1.64 mmol) was added to the mixture. The reaction mixture was hydrogenated using a hydrogen balloon at ambient temperature for 2 days. The mixture was filtered through diatomaceous earth, and the filter cake was washed with methanol and ethyl acetate. The filtrate was concentrated under reduced pressure to give the crude title compound. LC / MS (ESI+) m / z 307.2 (M+H)+. Example 9C (R)-4-(6-((6-(4-(1-(3-(tributyl)-1,2,4-diazol-5-methamido)ethyl)-3-methylphenyl)pyrimidin-4-yl)amino)pyridin-3-yl)-3,3-dimethylpiperazine-1-carboxylic acid tributyl ester

[0098] A stirred mixture of Example 9B (400 mg, 1.30 mmol), Example 1E (522 mg, 1.30 mmol), racemic BINAP-Pd-G3 (CAS 2151915-22-7, 130 mg, 0.13 mmol), and cesium carbonate (1276 mg, 3.92 mmol) in 1,4-dimethylamine (6.5 mL) was heated for 2 hours at 105 °C in a Biotage™ initiator + microwave synthesizer. The mixture was cooled and filtered. The filtrate was concentrated under reduced pressure and purified by silica chromatography with ethyl acetate and hexane to provide the title compound. LC / MS (ESI+): m / z 670.6 (M+H)+. Example 9D(R)-3-(tert-butyl)-N-(1-(4-(6-((5-(2,2-dimethylpiperazine-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)-2-methylphenyl)ethyl)-1,2,4-diazol-5-methylamine

[0099] Example 9C (845 mg, 1.26 mmol) was dissolved in 1,4-dimethylamine (3 mL), and the hydrochloride salt (4 N contained in 1,4-dimethylamine, 1.6 mL) was added to the mixture. The mixture was stirred at ambient temperature for 2 hours and concentrated under reduced pressure to give the crude title compound as the hydrochloride salt. LC / MS (ESI+): m / z 570.7 (M+H)+. Example 9E 3-Tributyl-N-{(1R)-1-[4-(6-{[5-(4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2,2-dimethylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine

[0100] Example 9D (200 mg, 0.35 mmol), Example 1J (145 mg, 0.42 mmol), N,N-diisopropylethylamine (123 µL, 0.70 mmol), and potassium iodide (291 mg, 1.76 mmol) were mixed in N,N-dimethylformamide (3.5 mL). The mixture was stirred at 80 °C for 4 hours. The mixture was cooled, quenched with a few drops of formic acid, and directly subjected to reversed-phase preparative HPLC to isolate the title compound (Phenomenex Gemini® AXA C18 column, 30 × 250 mm × 5 µm, 0.1% formic acid / water and acetonitrile as mobile phase). 1H NMR (500 MHz, dimethyl sulfoxide - d 6): δppm 10.25 (s, 1H), 10.07 (s, 1H), 9.81 (d, J = 7.8 Hz, 1H), 8.68 (d, J = 1.1 Hz, 1H), 8.04 (d, J = 3.0 Hz, 2H), 7.80-7.76 (m, 2H), 7.67 (d, J = 8.8 Hz, 1H), 7.54 (d, J = 8.7 Hz, 1H), 7.50 (dd, J = 8.9, 2.7 Hz, 1H), 7.21 (d, J = 8.6 Hz, 2H), 7.17 (d, J = 8.5 Hz, 2H), 5.27 (p, J = 7.1 Hz, 1H), 3.70 (t, J = 6.7 Hz, 2H), 3.01 (s, 2H), 2.70 (t, J = 7.6 Hz, 2H), 2.64 (td, J = 6.7, 2.6 Hz, 2H), 2.50 – 2.43 (m, 4H), 2.41 (s, 3H), 2.37-2.27 (m, 2H), 1.45 (d, J = 7.0 Hz, 3H), 1.29 (s, 9H), 0.94 (s, 6H). LC / MS (ESI+) m / z786.9 (M+H) +. Example 10 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-3-fluoro-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine Example 10A 1-Bromo-2-fluoro-4-iodo-3-methylbenzene

[0101] 1-Bromo-2-fluoro-4-iodobenzene (288 g, 0.96 mol) in anhydrous tetrahydrofuran (2.0 L) was added to a 5 L reactor. The mixture was cooled to -60 °C, and lithium diisopropylamine (2 M in tetrahydrofuran, 526 mL, 1.05 mol) was added, and the mixture was stirred at -60 °C for 1 hour. Iodomethane (163 g, 1.15 mol) was added, and the mixture was stirred at -60 °C for 12 hours. The reaction mixture was diluted with water (1.0 L) and extracted with methyl tributyl ether (1.0 L × 3). The same reaction was repeated three times on the same scale. The combined organic layers from the four batches were washed with brine, dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to provide the crude title compound. ¹H NMR (400 MHz, CDCl₃): δppm 7.46 (d, J=8.4 Hz, ¹H), 7.10 (t, J=7.6 Hz, ¹H), 2.41 (s, ³H) ppm. Example 10B: 4-Bromo-3-fluoro-2-methylbenzaldehyde

[0102] Example 10A (360 g, 1.14 mol) and anhydrous tetrahydrofuran (1.8 L) were added to a 5 L reactor and cooled to -60 °C. iPrMgCl (isopropylmagnesium chloride, 2 M contained in tetrahydrofuran, 572 mL, 1.14 mol) was added, and the mixture was stirred at -60 °C for 1 hour. Morpholine-4-carboxaldehyde (158 g, 1.37 mol) was added to the mixture at -60 °C. The mixture was stirred at 0–5 °C for 4 hours. The reaction mixture was diluted with saturated NH4Cl aqueous solution (0.8 L) and extracted with methyl tributyl ether (1.7 L × 3). The same reaction was repeated twice on the same scale. The combined organic layers from the three batches were washed with brine, dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to provide the crude title compound. ¹H NMR (400 MHz, CDCl₃): δppm 10.18 (s, ¹H), 7.54 (d, J=8.4 Hz, ¹H), 7.45 (d, J=8.4 Hz, ¹H), 2.60 (s, 3H) ppm. Example 10C(S, E)-N-(4-bromo-3-fluoro-2-methylbenzyl)-2-methylpropane-2-sulfinamide

[0103] At ambient temperature, triisopropyl borate (840 g, 4.47 mol), crude Example 10B (746 g, 3.44 mol), and (S)-2-methylpropane-2-sulfinamide (542 g, 4.47 mol) were added to a 3 L reactor. The mixture was stirred at 70 °C for 12 hours. The mixture was cooled to ambient temperature, diluted with water (3.0 L), extracted with dichloromethane (2.0 L × 2), washed with brine, dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was ground with methyl tributyl ether to provide the title compound. ¹H NMR (400 MHz, CDCl₃): δppm 8.78 (s, ¹H), 7.60 (d, J=8.4 Hz, ¹H), 7.49 (t, J=7.6 Hz, ¹H), 2.55 (s, ³H), 1.28 (s, 9H) ppm. MS (ESI) m / z 320.0, 322.0 (M+H) +. Example 10D(S)-N-((R)-1-(4-bromo-3-fluoro-2-methylphenyl)ethyl)-2-methylpropane-2-sulfinamide

[0104] Anhydrous dichloromethane (4.4 L) and Example 10C (220 g, 0.69 mol) were added to a 10 L reactor. The mixture was cooled to 0 °C. CH3MgBr (3.0 M contained in tetrahydrofuran, 573 L, 1.72 mol) was added dropwise at 0 °C. The mixture was then stirred at ambient temperature for 12 hours. The reaction mixture was cooled in an ice bath, and a saturated aqueous solution of NH4Cl (2.2 L) was added to the mixture. The organic phase was separated, and the aqueous phase was extracted with dichloromethane (2.2 L × 2). The combined organic layers were washed with brine, dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was ground in hexane, filtered, and dried under reduced pressure to provide the title compound. ¹H NMR (400 MHz, dimethyl sulfoxide): δppm 7.52 (t, J=8.0 Hz, 1H), 7.19 (d, J=8.8 Hz, 1H), 5.43 (d, J=5.6 Hz, 1H), 4.59 (m, 1H), 2.26 (s, 3H), 1.42 (d, J=6.8 Hz, 3H), 1.09 (s, 9H) ppm. MS (ESI) m / z 336.0, 338.0 (M+H)+. Example 10E(S)-N-(R)-1-(3-fluoro-2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethyl)-2-methylpropane-2-sulfinamide

[0105] At 20°C, bis(pinacol)diboron (9.13 g, 36.0 mmol) and potassium acetate (5.88 g, 60.0 mmol) were added to a solution of Example 10D (10.5 g, 30.0 mmol) in 200 mL of 1,4-dioxane. The reaction vessel was evacuated and backfilled with nitrogen three times, followed by the addition of a 1,1'-bis(diphenylphosphino)ferrocene-dichloropalladium(II)dichloromethane complex (2.45 g, 3.00 mmol). The reaction vessel was again evacuated and backfilled with nitrogen three times. The reaction mixture was heated to 100°C and stirred for 16 hours. The reaction mixture was cooled and filtered through a diatomaceous earth filter. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography, eluting with petroleum ether and ethyl acetate to provide the title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d6): δppm 7.38–7.48 (m, ¹H), 7.24 (d, J=7.75 Hz, ¹H), 5.35–5.43 (m, ¹H), 4.53–4.69 (m, ¹H), 2.19 (d, J=2.00 Hz, 3H), 1.42 (d, J=6.63 Hz, 3H), 1.28 (s, ¹²H), 1.08 (s, ⁹H) ppm. Example 10F(R)-1-(3-fluoro-2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethane-1-amine

[0106] At ambient temperature, hydrochloride (4 N contained in methyl tributyl ether, 400 mL) was added dropwise to a solution of Example 10E (20.0 g, 52.2 mmol) in methyl tributyl ether (400 mL). The mixture was stirred for 3 hours, concentrated under reduced pressure, and ground in methyl tributyl ether (50 mL). The solid was isolated by filtration and dried under vacuum to give the crude title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d⁶): δ 8.63 (br s, 3H), 7.56–7.49 (m, 1H), 7.43 (d, J = 7.9 Hz, 1H), 4.57 (q, J = 6.7 Hz, 1H), 2.22 (d, J = 1.8 Hz, 3H), 1.46 (d, J = 6.6 Hz, 3H), 1.29 (s, 12H) ppm. MS (ESI) m / z 280.0 (M+H)⁺. Example 10G (R)-3-(tert-butyl)-N-(1-(3-fluoro-2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethyl)-1,2,4-diazol-5-methylamine

[0107] At 20°C, triethylamine (23.5 mL, 169 mmol) was added to a solution of Example 10F (7.00 g, 21.1 mmol) and methyl 3-(tributyl)-1,2,4-diazol-5-carboxylate (9.70 g, 52.7 mmol) in ethanol (140 mL). The mixture was stirred at 60°C for 12 hours, followed by concentration under reduced pressure to remove ethanol. The crude material was purified by silicone column chromatography, eluted with petroleum ether and ethyl acetate to provide the title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d⁶): δppm 9.90 (br d, J = 7.6 Hz, ¹H), 7.47 (t, J = 6.9 Hz, ¹H), 7.38–7.22 (m, ¹H), 5.28 (br t, J = 7.2 Hz, ¹H), 2.33–2.18 (m, 3H), 1.46 (br d, J = 6.9 Hz, 3H), 1.42–1.19 (m, 2¹H) ppm. MS (ESI) m / z 432.2 (M+H)⁺. Example 10H(R)-3-(tert-butyl)-N-(1-(4-(6-chloropyrimidinyl-4-yl)-3-fluoro-2-methylphenyl)ethyl)-1,2,4-diazol-5-methamide

[0108] At 20°C, a solution of potassium phosphate (2.76 g, 12.98 mmol) in water (30 mL) was added to a mixture of Example 10 G (2.80 g, 6.49 mmol) and 4,6-dichloropyrimidine (1.93 g, 12.98 mmol) in tetrahydrofuran (120 mL). The mixture was degassed under a stream of nitrogen at ambient temperature, and [1,1'-bis(diphenylphosphine)ferrocene]dichloropalladium(II) (265 mg, 0.33 mmol) was added to the mixture under N2. The reaction mixture was heated to 80°C and stirred for 2 hours. The mixture was cooled, poured into water, and extracted with ethyl acetate (three times). The combined organic layers were washed with brine, dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica column chromatography, eluted with tetrahydrofuran / petroleum ether to provide the crude title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d⁶): δppm 9.93 (d, J=7.8 Hz, ¹H), 9.07 (s, ¹H), 8.30 (s, ¹H), 8.09 (d, J=7.8 Hz, ¹H), 7.63 (d, J=8.5 Hz, ¹H), 5.33 (quin, J=7.2 Hz, ¹H), 2.48–2.47 (m, ¹H), 2.47 (s, 2H), 1.50 (d, J=7.0 Hz, 3H), 1.36 (s, 9H) ppm. MS (ESI) m / z 418.2 (M+H)⁺. Example 10I (S)-4-(6-((6-(4-((R)-1-(3-(tributyl)-1,2,4-diazol-5-methamido)ethyl)-2-fluoro-3-methylphenyl)pyrimidin-4-yl)amino)pyridin-3-yl)-3-methylpiperazol-1-carboxylic acid tributyl ester

[0109] To a solution of Example 10H (500 mg, 1.20 mmol) in 1,4-dimethylamine (10 mL), add Example 1L (385 mg, 1.32 mmol), Pd₂(dba)₃ (CAS 52409-22-0, 55 mg, 0.06 mmol), XantPhos (CAS 161265-03-8, 69 mg, 0.12 mmol), and cesium carbonate (780 mg, 2.39 mmol). Degas the mixture using a nitrogen stream and stir at 100 °C for 16 hours under a nitrogen atmosphere. Cool the reaction mixture, dilute with ethyl acetate, filter through diatomaceous earth, concentrate under reduced pressure, and purify by silica chromatography using ethyl acetate and petroleum ether to give the title compound. LC / MS (ESI+): m / z 674.4 (M+H)⁺. Example 10J 3-(tert-butyl)-N-((R)-1-(3-fluoro-2-methyl-4-(6-((5-((S)-2-methylpiperazine-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)phenyl)ethyl)-1,2,4-diazol-5-methylamine

[0110] The hydrochloride salt (4 N contained in methyl tributyl ether, 2 mL) was added dropwise to a solution of Example 10I (150 mg, 0.223 mmol) in methyl tributyl ether (2 mL). The reaction mixture was stirred at ambient temperature for 2 hours. The reaction mixture was concentrated under reduced pressure to give the title compound as the hydrochloride salt. LC / MS (ESI+): m / z 574.4 (M+H)+. Example 10K 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-3-fluoro-2-methylphenyl]ethyl]-1,2,4-diazol-5-methylamine

[0111] Example 10J (100 mg, 0.17 mmol) was added to a solution of N,N-dimethylformamide (2 mL) containing Example 1J (54 mg, 0.17 mmol), potassium iodide (145 mg, 0.87 mmol), and potassium carbonate (48 mg, 0.35 mmol). The mixture was stirred at 80 °C for 2 hours. The mixture was cooled to ambient temperature, filtered, and subjected directly to reversed-phase preparative HPLC to isolate the title compound (Gilson GX-281 system, Phenomenex Gemini® NX C18 column, 30 × 75 mm × 3 µm, 10 mM ammonium bicarbonate / water and acetonitrile as mobile phase). 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 10.33 (s, 1H), 10.08 (s, 1H), 9.93 (br d, J = 7.5 Hz, 1H), 8.72 (s, 1H), 7.96 (br d, J = 2.4 Hz, 2H), 7.81 (t, J = 8.0 Hz, 1H), 7.72 (br d, J = 2.0 Hz, 1H), 7.43 (br d, J = 8.3 Hz, 2H), 7.32-7.17 (m, 4H), 5.33 (quin, J = 6.9 Hz, 1H), 3.92-3.83 (m, 1H), 3.76 (t, J = 6.7 Hz, 2H), 3.26-3.16 (m, 1H), 2.98 (br t, J = 9.5 Hz, 1H), 2.85 (br d, J = 10.6 Hz, 1H), 2.81-2.74 (m, 2H), 2.69 (br t, J = 6.7 Hz, 3H), 2.61 - 2.54 (m, 2H), 2.46-2.42 (m, 1H), 2.35 (d, J = 1.8 Hz, 3H), 2.31-2.25 (m, 1H), 1.52 (d, J = 7.0 Hz, 3H), 1.36 (s, 9H), 0.98 (d, J = 6.3 Hz, 3H) ppm. MS (ESI) m / z790.3 (M+H) +.Example 11 3-Tributyl-N-{(1R)-1-[4-(6-{[5-(4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}piperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-3-fluoro-2-methylphenyl}ethyl]-1,2,4-diazol-5-carboxylic acid Example 11A (R)-4-(6-((6-(4-(1-(3-(tributyl)-1,2,4-diazol-5-carboxylic acid)ethyl)-2-fluoro-3-methylphenyl)pyrimidin-4-yl)amino)pyridin-3-yl)-piperazine-1-carboxylic acid tributyl ester.

[0112] To a solution of Example 10H (2.40 g, 5.46 mmol) in 1,4-dimethylamine (80 mL), Example 3B (1.67 g, 6.00 mmol), Pd₂(dba)₃ (CAS 52409-22-0, 250 mg, 0.27 mmol), XantPhos (CAS 161265-03-8, 316 mg, 0.32 mmol) and cesium carbonate (3.56 g, 10.91 mmol) were added. The mixture was degassed using a nitrogen stream and stirred at 100 °C for 16 hours under a nitrogen atmosphere. The reaction mixture was cooled, diluted with ethyl acetate, filtered through diatomaceous earth, concentrated under reduced pressure, and purified by silica chromatography using ethyl acetate and petroleum ether to give the title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d⁶): δppm 10.15 (s, ¹H), 9.96 (d, J = 7.8 Hz, ¹H), 8.75 (s, ¹H), 8.08 - 7.94 (m, 2H), 7.87 - 7.68 (m, 2H), 7.52 - 7.40 (m, 2H), 5.41 - 5.29 (m, 1H), 3.48 (br d, J = 4.8 Hz, 4H), 3.14 - 3.03 (m, 4H), 2.37 (d, J = 1.9 Hz, 3H), 1.53 (d, J = 7.0 Hz, 3H), 1.43 (s, 9H), 1.38 (s, 9H) ppm. LC / MS (ESI+): m / z 660.4 (M+H)+. Example 11B (R)-3-(tributyl)-N-(1-(3-fluoro-2-methyl-4-(6-((5-(piperidin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)phenyl)ethyl)-1,2,4-diazol-5-methylamine

[0113] The hydrochloride salt (4 N contained in 1,4-dimethylamine, 40 mL) was added dropwise to a solution of Example 11A (3.15 g, 4.30 mmol) in 1,4-dimethylamine (40 mL). The reaction mixture was stirred at ambient temperature for 2 hours. The reaction mixture was concentrated under reduced pressure to give the title compound as the hydrochloride salt. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 10.11 (s, 1H), 10.06 - 9.88 (m, 1H), 8.74 (s, 1H), 8.00 (br s, 2H), 7.87 - 7.64 (m, 2H), 7.45 (br d, J= 7.1 Hz, 2H), 5.35 (br t, J= 6.9 Hz, 1H), 3.04 (br s, 4H), 2.86 (br s, 4H), 2.37 (br s, 3H), 1.53 (br d, J= 6.6 Hz, 3H), 1.37 (s, 9H)ppm. LC / MS (ESI+): m / z 560.3 (M+H)+. Example 11C 3-Tributyl-N-{(1R)-1-[4-(6-{[5-(4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}piperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-3-fluoro-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine

[0114] Example 1J (86 mg, 0.25 mmol), sodium bromide (129 mg, 1.25 mmol), and sodium bicarbonate (70 mg, 0.84 mmol) were added to a solution of N,N-dimethylformamide (1 mL) containing Example 11B (130 mg, 0.21 mmol). The mixture was stirred at 80 °C for 12 hours. The mixture was cooled to ambient temperature, filtered, and subjected directly to reversed-phase preparative HPLC to isolate the title compound (Phenomenex Gemini® NX C18 column, 40 × 80 mm × 3 µm, 10 mM ammonium bicarbonate / water and acetonitrile as mobile phase). 1H NMR (400 MHz, dimethyl sulfoxide-d 6): δppm 10.34 (s, 1 H), 10.09 (s, 1 H), 9.94 (br d, J=7.63 Hz, 1 H), 8.72 (s, 1 H), 8.01 (br s, 1 H), 7.81 (br t, J=8.07 Hz, 1 H), 7.39-7.49 (m, 2 H), 7.21-7.30 (m, 3 H), 5.33 (br t, J=6.94 Hz, 1 H), 3.76 (t, J=6.63 Hz, 2 H), 3.14 (br s, 5 H), 2.74-2.82 (m, 2 H), 2.69 (br t, J=6.69 Hz, 2 H), 2.52-2.64 (m, 6 H), 2.35 (d, J=1.50 Hz, 3 H), 1.51 (br d, J=6.88 Hz, 3 H), 1.36 (s, 9 H) ppm. MS (ESI) m / z776.3 (M+H) +. Example 12 3-Tributyl-N-{(1R)-1-[4-(6-{[5-(4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}piperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-5-fluoro-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine Example 12A 4-Bromo-5-fluoro-2-methylbenzaldehyde

[0115] 1-Bromo-2-fluoro-4-iodo-5-methylbenzene (25 g, 79 mmol) and anhydrous tetrahydrofuran (400 mL) were added to a 2 L flask and cooled to -25 °C. iPrMgCl (isopropylmagnesium chloride, 1.3 M contained in tetrahydrofuran, 67.2 mL, 87 mmol) was added, and the mixture was stirred at -25 °C for 30 min. Anhydrous N,N-dimethylformamide (7.38 mL, 96 mmol) was added to the mixture at -25 °C. The mixture was heated to 0-5 °C and stirred for 1.5 h. The reaction mixture was carefully quenched with 1 N hydrochloric acid (200 mL), and the mixture was extracted with ethyl acetate (three times). The combined organic phases were dried over sodium sulfate, filtered, and concentrated under reduced pressure to give the crude title compound. ¹H NMR (400 MHz, CDCl₃): δppm 10.21 (d, J=1.7 Hz, ¹H), 7.58–7.54 (m, 2H), 2.64 (s, 3H) ppm. Example 12B (S, E)-N-(4-bromo-5-fluoro-2-methylbenzyl)-2-methylpropane-2-sulfinamide

[0116] Example 12A (17.00 g, 78.0 mmol) and dichloromethane (340 mL) were added to a 1 L flask. At ambient temperature, (S)-2-methylpropane-2-sulfinamide (10.44 g, 86.0 mmol) and titanium ethoxide (IV) (35.7 g, 157 mmol) were added to this stirred solution. The mixture was stirred for 16 hours and then quenched with water (36 mL) at 0 °C. The mixture was filtered, and the filter cake was washed with dichloromethane (300 mL three times). The filtrate was concentrated under reduced pressure, and the crude material was ground in hexane (50 mL) for 30 minutes. The solid was isolated by filtration, washed with hexane, and dried under high vacuum to give the title compound. Example 12C(S)-N-((R)-1-(4-bromo-5-fluoro-2-methylphenyl)ethyl)-2-methylpropane-2-sulfinamide

[0117] Add anhydrous dichloromethane (400 mL) and Example 12B (21.6 g, 67.5 mmol) to a 2 L flask. Cool the mixture to 0 °C. Add CH3MgBr (3.0 M in tetrahydrofuran, 56.2 L, 169 mmol) dropwise at 0 °C. Then, stir the mixture at 0 °C for 5 hours. Quench the reaction mixture with saturated NH4Cl aqueous solution (500 mL). Separate the organic phase and extract the aqueous phase with dichloromethane (100 mL × 3). Wash the combined organic layers with brine, dry with sodium sulfate, and filter. Concentrate the filtrate under reduced pressure and purify the residue by silica column chromatography using ethyl acetate and petroleum ether to isolate the title compound. MS (ESI) m / z 335.9, 337.9 (M+H)+. Example 12D(S)-N-(R)-1-(5-fluoro-2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethyl)-2-methylpropane-2-sulfinamide

[0118] At 20°C, bis(pinacolyl)diboron (13.19 g, 51.9 mmol) and potassium acetate (8.49 g, 87.0 mmol) were added to a solution of Example 12C (15.0 g, 43.3 mmol) in 1,4-dioxane (300 mL). The reaction vessel was evacuated and backfilled with nitrogen three times, followed by the addition of a 1,1'-bis(diphenylphosphino)ferrocene-dichloropalladium(II)dichloromethane complex (3.53 g, 4.33 mmol). The reaction vessel was again evacuated and backfilled with nitrogen three times. The reaction mixture was heated to 100°C and stirred for 16 hours. The reaction mixture was cooled and filtered through a diatomaceous earth filter. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography, eluting with petroleum ether and ethyl acetate to provide the title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d6): δppm 7.51 (d, J=6.0 Hz, 1H), 7.05 (d, J=10.5 Hz, 1H), 4.80 (br d, J=3.4 Hz, 1H), 3.49 (br d, J=2.6 Hz, 1H), 2.34 (s, 3H), 1.50–1.44 (m, 3H), 1.36 (s, 12H), 1.23–1.19 (m, 9H) ppm. Example 12E(R)-1-(5-fluoro-2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethane-1-amine

[0119] At ambient temperature, hydrochloride (4 N contained in 1,4-dimethylethane, 30 mL) was added dropwise to a solution of Example 12D (3.00 g, 7.83 mmol) in dichloromethane (10 mL). The mixture was stirred for 24 hours and concentrated under reduced pressure to give the crude title compound. MS (ESI) m / z 280.0 (M+H)+. Example 12F (R)-3-(tributyl)-N-(1-(5-fluoro-2-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethyl)-1,2,4-diazol-5-methylamine

[0120] At ambient temperature, 2,3,4,6,7,8,9,10-octahydropyrimidine[1,2-a]azine (6.16 mL, 40.80 mmol) was added to a solution of Example 12E (3.80 g, 13.61 mmol) and methyl 3-(tributyl)-1,2,4-diazol-5-carboxylate (3.76 g, 20.42 mmol) in tetrahydrofuran (60 mL). The reaction mixture was stirred at 85 °C for 16 hours, cooled, concentrated under reduced pressure, and purified by silicone column chromatography, eluting with hexane and ethyl acetate to provide the title compound. MS (ESI) m / z 432.2 (M+H)+. Example 12G(R)-3-(tert-butyl)-N-(1-(4-(6-chloropyrimidinyl-4-yl)-5-fluoro-2-methylphenyl)ethyl)-1,2,4-diazol-5-methamide

[0121] The title compound was prepared according to the procedure used for Example 10H, replacing Example 10G with Example 12F. MS (ESI) m / z 418.2 (M+H)+. Example 12H (R)-4-(6-((6-(4-(1-(3-(tributyl)-1,2,4-diazol-5-methamido)ethyl)-2-fluoro-5-methylphenyl)pyrimidin-4-yl)amino)pyridin-3-yl)-piperazine-1-carboxylic acid tributyl ester

[0122] At 110 °C under a nitrogen atmosphere, a stirred mixture of Example 3B (406 mg, 1.46 mmol), Example 12G (435 mg, 1.04 mmol), racemic BINAP-Pd-G3 (CAS 2151915-22-7, 99 mg, 0.1 mmol), and cesium carbonate (746 mg, 2.29 mmol) in 6.5 mL of 1,4-dimethylamine was heated for 4 hours. The mixture was cooled, filtered, concentrated under reduced pressure, and the residue was purified by silica chromatography with ethyl acetate and hexane to isolate the title compound. LC / MS (ESI+): m / z 660.4 (M+H)+. Example 12I (R)-3-(tert-butyl)-N-(1-(5-fluoro-2-methyl-4-(6-((5-(piperidin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)phenyl)ethyl)-1,2,4-diazol-5-methylamine

[0123] The title compound was prepared according to the procedure used in Example 11B, replacing Example 11A with Example 12H. MS (ESI) m / z 560.4 (M+H) +. Example 12J 3-Tributyl-N-{(1R)-1-[4-(6-{[5-(4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}piperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-5-fluoro-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine

[0124] Example 1J (81 mg, 0.24 mmol), potassium iodide (139 mg, 0.84 mmol), and N,N-diisopropylethylamine (205 µL, 1.17 mmol) were added to a solution of Example 12I (100 mg, 0.17 mmol) in N,N-dimethylformamide (1.7 mL). The mixture was stirred at 80 °C for 2 hours. The mixture was cooled to ambient temperature, filtered, and directly subjected to reversed-phase preparative HPLC to separate the title compound (Phenomenex Gemini® AXA C18 column, 30 × 250 mm × 5 µm, 0.1% formic acid / water and acetonitrile as mobile phase). 1H NMR (500 MHz, dimethyl sulfoxide - d 6): δppm 10.34 (s, 1H), 10.11 (s, 1H), 9.87 (d, J= 7.9 Hz, 1H), 8.73 (s, 1H), 8.04-7.95 (m, 2H) 7.83 (d, J= 8.0 Hz, 1H), 7.72 (brs, 1H), 7.50-7.45 (m, 1H), 7.43 (d, J= 12.5 Hz, 1H), 7.30-7.20 (m, 4H), 5.29 (p, J= 7.2 Hz, 1H), 3.76 (t, J= 6.7 Hz, 2H), 3.14 (brs, 4H), 2.81 (brs, 2H), 2.70 (t, J= 6.8 Hz, 2H), 2.67-2.55 (m, 3H), 2.42 (s, 3H), 1.51 (d, J= 7.0 Hz, 3H), 1.37 (s, 9H) ppm. MS (ESI) m / z776.8 (M+H) +. Example 13 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-(propyl-2-yl)piperidin-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-carboxamide Example 13A (S)-3-isopropyl-4-(6-nitropyridin-3-yl)piperidin-1-carboxylic acid tributyl ester

[0125] At 20°C under N2, tributyl (S)-3-isopropylpiperazine-1-carboxylic acid (500 mg, 2.190 mmol) and N,N-diisopropylethylamine (1.147 mL, 6.57 mmol) were added to a mixture of 5-fluoro-2-nitropyridine (467 mg, 3.28 mmol) in dimethyl sulfoxide (10 mL). The reaction mixture was stirred at 120°C for 16 hours. The mixture was cooled and poured into ice water (30 mL). The material in the mixture was collected by filtration. The material was milled in a mixture of ethyl acetate and petroleum ether to form a slurry. The slurry was filtered, and the filter cake was dried under vacuum to provide the title compound. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 8.25 (d, J=3.00 Hz, 1 H), 8.11 (d, J=9.26 Hz, 1 H), 7.46 (dd, J=9.38, 3.00 Hz, 1 H), 4.02-4.13 (m, 1 H), 3.92 (br d, J=12.26 Hz, 3 H), 3.18 (br s, 1 H), 2.84-3.10 (m, 2 H), 2.01-2.15 (m, 1 H), 1.41 (s, 9 H), 1.05 (br d, J=4.38 Hz, 3 H), 0.73 (d, J=6.75 Hz, 3 H) ppm. Example 13B (S)-4-(6-aminopyridin-3-yl)-3-isopropylpiperazine-1-carboxylic acid tributyl ester

[0126] At ambient temperature, Example 13A (400 mg, 1.142 mmol) was added to a mixture of Pd / C (10 wt%, 121 mg, 0.114 mmol) in ethyl acetate (12 mL). The mixture was hydrogenated using a hydrogen balloon for 2 hours. The mixture was filtered through a diatomaceous earth pad, and the filtrate was concentrated under reduced pressure to provide the crude title compound. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 7.66 (br s, 1 H), 7.21 (br d, J=7.75 Hz, 1 H), 6.39 (d, J=8.75 Hz, 1 H), 5.49 (br s, 2 H), 3.35-3.65 (m, 2 H), 2.90 (br s, 3 H), 2.51-2.54 (m, 2 H), 1.77 (br s, 1 H), 1.41 (s, 9 H), 0.70 - 0.91 (m, 6 H) ppm. Example 13C(S)-4-(6-((6-(4-((R)-1-(3-(tributyl)-1,2,4-diazol-5-methamido)ethyl)-3-methylphenyl)pyrimidin-4-yl)amino)pyridin-3-yl)-3-isopropylpiperazine-1-carboxylic acid tributyl ester

[0127] To a solution of Example 1M (200 mg, 0.49 mmol) in 1,4-dimethylamine (5 mL), Example 13B (233 mg, 0.73 mmol), Pd₂(dba)₃ (CAS 52409-22-0, 44 mg, 0.049 mmol), XantPhos (CAS 161265-03-8, 28 mg, 0.049 mmol) and cesium carbonate (316 mg, 0.97 mmol) were added. The mixture was degassed using a nitrogen stream and stirred at 100 °C for 12 hours. The reaction mixture was cooled, diluted with ethyl acetate, and filtered through diatomaceous earth. The filtrate was concentrated under reduced pressure, and the residue was purified by silica chromatography using tetrahydrofuran and petroleum ether to give the title compound. LC / MS (ESI+): m / z 684.5 (M+H)⁺. Example 13D 3-(tert-butyl)-N-((R)-1-(4-(6-((5-((S)-2-isopropylpiperazol-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)-2-methylphenyl)ethyl)-1,2,4-diazol-5-methylamine

[0128] At ambient temperature, the hydrochloride (4N contained in diane, 3 mL) of Example 13C (280 mg, 0.372 mmol) was added dropwise to a solution in 1,4-diane (3 mL). The mixture was stirred for 1 hour and concentrated under reduced pressure to give the crude title compound as hydrochloride. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δ ppm 9.89 (s, 1 H), 8.67 (s, 1 H), 7.97 (br d, J=2.64 Hz, 2 H), 7.77 - 7.88 (m, 1 H), 7.74 - 7.89 (m, 1 H), 7.55 - 7.69 (m, 2 H), 7.21 - 7.43 (m, 2 H), 5.33 (quin, J=7.12 Hz, 1 H), 3.21 - 3.31 (m, 3 H), 3.00 - 3.11 (m, 1 H), 2.83 - 2.92 (m, 1 H), 2.62 - 2.82 (m, 3 H), 2.47 (s, 3 H), 1.51 (d, J=6.90 Hz, 3 H), 1.36 (s, 9 H), 0.89 (d, J=6.53 Hz, 3 H), 0.75 (d, J=6.78 Hz, 3 H) ppm. LC / MS (ESI+): m / z584.5 (M+H) +. Example 13E 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-(propyl-2-yl)piperidin-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine

[0129] Example 1J (175 mg, 0.51 mmol), sodium iodide (153 mg, 1.02 mmol), and N,N-diisopropylethylamine (296 µL, 1.70 mmol) were added to a solution of Example 13D (220 mg, 0.34 mmol) in acetonitrile (5 mL). The mixture was stirred at 80 °C for 16 hours. The mixture was cooled to ambient temperature, filtered, and subjected directly to reversed-phase preparative HPLC to isolate the title compound (Waters™ Xbridge C18 column, 30 × 100 mm × 10 µm, 10 mM ammonium bicarbonate / water and acetonitrile as mobile phase). 1H NMR (400 MHz, dimethyl sulfoxide-d 6): δppm 10.35 (s, 1 H), 9.86-10.00 (m, 2 H), 8.68 (s, 1 H), 8.00 (br d, J=2.63 Hz, 2 H), 7.82 (br s, 2 H), 7.60 (br d, J=8.55 Hz, 2 H), 7.36-7.47 (m, 1 H), 7.25 (q, J=8.70 Hz, 4 H), 5.27-5.39 (m, 1 H), 3.69-3.80 (m, 2 H), 3.38-3.43 (m, 2 H), 3.09-3.20 (m, 1 H), 2.88 (br s, 1 H), 2.73-2.79 (m, 2 H), 2.67-2.71 (m, 2 H), 2.52 (d, J=1.75 Hz, 2 H), 2.47 (s, 3 H), 2.28-2.35 (m, 2 H), 2.17-2.26 (m, 2 H), 1.51 (d, J=7.02 Hz, 3 H), 1.36 (s, 9 H), 0.89 (d, J=6.58 Hz, 3 H), 0.76 (d, J=6.80 Hz, 3 H) ppm. MS (ESI) m / z800.5 (M+H) +. Example 14 3-Tributyl-N-[(1R)-1-{2-chloro-4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]phenyl}ethyl]-1,2,4-diazol-5-methylamine Example 14A (S, E)-N-(4-bromo-2-chlorobenzyl)-2-methylpropane-2-sulfinamide

[0130] Triisopropyl borate (139 g, 0.74 mol), 4-bromo-2-chlorobenzaldehyde (125 g, 0.57 mol), and (S)-2-methylpropane-2-sulfinamide (90 g, 0.74 mol) were added to a 1 L flask at ambient temperature. The mixture was stirred at 70 °C for 12 hours. Water (0.5 L) was added, and the mixture was extracted with dichloromethane (0.5 L × 3), washed with brine (1.0 L), dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure. The reaction was repeated three times on the same scale. The crude materials from all four batches were combined to provide the title compound. MS (ESI) m / z 322.0, 324.0 (M+H)+. Example 14B (S)-N-((R)-1-(4-bromo-2-chlorophenyl)ethyl)-2-methylpropane-2-sulfinamide

[0131] Dichloromethane (3.5 L) and Example 14A (173 g, 0.53 mol) were added to a 5 L reactor. The mixture was cooled at 0 °C. CH3MgBr (3.0 M contained in tetrahydrofuran, 268 L, 0.80 mol) was added dropwise at 0 °C. The mixture was stirred at ambient temperature for 12 hours. The reaction mixture was cooled in an ice bath, and a saturated aqueous solution of NH4Cl (1.6 L) was added to the mixture. The organic phase was separated, and the aqueous phase was extracted with dichloromethane (0.5 L × 3). The reaction was repeated four times on the same scale. The combined organic layers were washed with brine (5.5 L), dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was ground in hexane to provide the crude title compound. MS (ESI) m / z 338.0, 340.0 (M+H)+. Example 14C(S)-N-((R)-1-(2-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethyl)-2-methylpropane-2-sulfinamide

[0132] At 20°C, bis(pinacolyl)diboron (10.1 g, 40.0 mmol) and potassium acetate (7.82 g, 80.0 mmol) were added to a solution of Example 14B (15.0 g, 40.0 mmol) in 1,4-dioxane (150 mL). The reaction vessel was evacuated and backfilled with nitrogen three times, and a 1,1'-bis(diphenylphosphino)ferrocene-dichloropalladium(II)dichloromethane complex (3.26 g, 4.0 mmol) was added. The reaction vessel was again evacuated and backfilled with nitrogen three times. The reaction mixture was heated to 110°C and stirred for 16 hours. The reaction mixture was cooled and filtered through a diatomaceous earth filter. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography, eluted with petroleum ether and ethyl acetate to provide the title compound. MS (ESI) m / z 386.2 (M+H) +. Example 14D(R)-1-(2-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethane-1-amine

[0133] At ambient temperature, the hydrochloride salt (4N contained in 1,4-dimethylbenzane, 100 mL) was added dropwise to a solution of Example 14C (8.00 g, 20.7 mmol) in 1,4-dimethylbenzane (100 mL). The mixture was stirred for 2 hours and concentrated under reduced pressure to give the crude title compound as the hydrochloride salt. MS (ESI) m / z 282.3 (M+H)+. Example 14E (R)-3-(tributyl)-N-(1-(2-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethyl)-1,2,4-diazol-5-methylamine

[0134] At 20°C, triethylamine (17.4 mL, 125 mmol) was added to a solution of Example 14D (6.60 g, 20.8 mmol) and methyl 3-(tributyl)-1,2,4-diazol-5-carboxylate (7.64 g, 41.5 mmol) in ethanol (180 mL). The mixture was stirred at 80°C for 12 hours, followed by concentration under reduced pressure to remove ethanol. The crude material was purified by silicone column chromatography, eluted with petroleum ether and ethyl acetate to provide the title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d6): δppm 9.95 (d, J=7.5 Hz, 1H), 7.65–7.56 (m, 3H), 5.42 (quin, J=7.2 Hz, 1H), 1.48 (d, J=7.1 Hz, 3H), 1.36 (s, 9H), 1.28 (s, 12H) ppm. MS (ESI) m / z 434.4 (M+H)+. Example 14F(R)-3-(tributyl)-N-(1-(4-(6-chloropyrimidinyl-4-yl)-2-chlorophenyl)ethyl)-1,2,4-diazol-5-methylamine

[0135] [1,1'-bis(diphenylphosphine)ferrocene]dichloropalladium(II) (1.13 g, 1.38 mmol) and potassium phosphate (5.87 g, 27.7 mmol) were added to a mixture of Example 14E (6.00 g, 13.83 mmol) and 4,6-dichloropyrimidine (2.47 g, 16.60 mmol) in 1,4-dimethylamine (111 mL) and water (28 mL). The mixture was degassed with a stream of nitrogen at ambient temperature and stirred at 80 °C for 1 hour under a nitrogen atmosphere. The mixture was cooled, poured into water, and extracted with ethyl acetate (three times). The combined organic layers were washed with brine, dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silicon column chromatography, eluted with ethyl acetate / hexane, to give the crude title compound. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 10.02 (br d, J=7.63 Hz, 1H), 9.11 (d, J=0.63 Hz, 1H), 8.39 (d, J=0.75 Hz, 1H), 8.31 (d, J=1.63 Hz, 1H), 8.23 ​​(dd, J=8.25, 1.63 Hz, 1H), 7.74 (d, J=8.25 Hz, 1H), 5.47 (br t, J=7.19 Hz, 1H), 1.53 (d, J=7.00 Hz, 3H), 1.37 (s, 9H) ppm. MS (ESI) m / z 420.1, 422.1 (M+H) +. Example 14G (S)-4-(6-((6-(4-((R)-1-(3-(tributyl)-1,2,4-diazol-5-methamido)ethyl)-3-phenylchloro)pyrimidin-4-yl)amino)pyridin-3-yl)-3-methylpiperazine-1-carboxylic acid tributyl ester

[0136] To a solution of Example 14H (288 mg, 0.69 mmol) in 1,4-dimethylamine (6.4 mL), Example 1L (240 mg, 0.82 mmol), Pd₂(dba)₃ (CAS 52409-22-0, 38 mg, 0.04 mmol), XantPhos (CAS 161265-03-8, 40 mg, 0.07 mmol), and cesium carbonate (447 mg, 1.37 mmol) were added. The mixture was degassed using a nitrogen stream and stirred at 100 °C for 16 hours under a nitrogen atmosphere. The reaction mixture was cooled, diluted with ethyl acetate, filtered through diatomaceous earth, concentrated under reduced pressure, and purified by silica chromatography using ethyl acetate and petroleum ether to give the title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d⁶): δppm 9.97–10.11 (m, 2H), 7.92–8.11 (m, 4H), 7.72 (d, J=8.11 Hz, 2H), 7.44 (dd, J=9.12, 2.80 Hz, 1H), 5.46 (m, 1H), 3.75–3.89 (m, 2H), 3.61 (br d, J=11.09 Hz, 1H), 3.17 (br d, J=11.80 Hz, 3H), 2.90–3.00 (m, 1H), 1.53 (d, J=7.03 Hz, 3H), 1.41 (s, 9H), 1.37 (s, 9H), 0.88 (d, J=6.32 Hz, 3H) ppm. LC / MS (ESI+): m / z 676.4 (M+H)+. Example 14H 3-(tributyl)-N-((R)-1-(2-chloro-4-(6-((5-((S)-2-methylpiperazol-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)phenyl)ethyl)-1,2,4-diazol-5-methylamine

[0137] Hydrochloride (4N contained in 1,4-dimethylamine, 3 mL) was added dropwise to a solution of Example 14 G (258 mg, 0.38 mmol) in 1,4-dimethylamine (3 mL). The reaction mixture was stirred at ambient temperature for 1 hour. The reaction mixture was concentrated under reduced pressure, dissolved in water (10 mL), and alkalized with a saturated aqueous sodium bicarbonate solution to adjust the pH to 8. The mixture was extracted with dichloromethane (10 mL, three times), and the combined organic phases were washed with brine (10 mL), dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give the title compound. 1H NMR (400 MHz, dimethyl sulfoxide-d 6): δppm 10.08 (br s, 2H), 8.72 (s, 1H), 7.91-8.10 (m, 4H), 7.71 (br d, J=8.13 Hz, 2H), 7.40 (br dd, J=9.13, 2.75 Hz, 2H), 5.46 (br s, 1H), 3.64-3.80 (m, 1H), 3.01-3.15 (m, 1H), 2.79-2.98 (m, 3H), 2.62-2.77 (m, 2H), 1.53 (br d, J=6.88 Hz, 3H), 1.36 (s, 9H), 0.95 (d, J=6.38 Hz, 3H) ppm. LC / MS (ESI+): m / z 576.3 (M+H)+. Example 14I 3-Tributyl-N-[(1R)-1-{2-chloro-4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]phenyl}ethyl]-1,2,4-diazol-5-methylamine

[0138] To a solution of Example 14H (85 mg, 0.15 mmol) in acetonitrile (2 mL), Example 1J (102 mg, 0.30 mmol), sodium iodide (66 mg, 0.44 mmol), and N,N-diisopropylethylamine (129 µL, 0.74 mmol) were added. The mixture was stirred at 80 °C for 16 hours. The mixture was cooled to ambient temperature, filtered, and subjected directly to reversed-phase preparative HPLC to isolate the title compound (Phenomenex Luna® C18 column, 30 × 75 mm × 3 µm, 0.1% formic acid / water and acetonitrile as mobile phase). 1H NMR (400 MHz, dimethyl sulfoxide-d 6): δppm 10.34 (s, 1 H), 9.97-10.13 (m, 2 H), 8.73 (s, 1 H), 7.91-8.11 (m, 4 H), 7.72 (br d, J=8.13 Hz, 2 H), 7.47 (br d, J=8.00 Hz, 1 H), 7.20-7.33 (m, 4 H), 5.46 (quin, J=7.10 Hz, 1 H), 3.80 - 3.95 (m, 1 H), 3.76 (t, J=6.63 Hz, 2 H), 3.16-3.26 (m, 2 H), 2.95-3.10 (m, 2 H), 2.57-2.93 (m, 8 H), 1.53 (d, J=7.00 Hz, 3 H), 1.37 (s, 9 H), 1.00 (br d, J=6.00 Hz, 3 H) ppm. MS (ESI) m / z792.2 (M+H) +. Example 15 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2R)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-(methoxymethyl)piperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine Example 15A (R)-2-(hydroxymethyl)piperazine-1,4-dicarboxylic acid 1-benzyl 4-tributyl ester

[0139] At 20 °C, a solution of sodium carbonate (2.205 g, 20.81 mmol) in water (6.00 mL) was added to a solution of tributyl (R)-3-(hydroxymethyl)piperazine-1-carboxylate (1.5 g, 6.94 mmol) in tetrahydrofuran (30 mL). Benzyl chloroformate (1.775 g, 10.40 mmol) was added dropwise at 0 °C. The reaction mixture was extracted with ethyl acetate (three times), washed with brine, dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography using ethyl acetate / petroleum ether to provide the title compound. 1H NMR (400 MHz, dimethyl sulfoxide-d 6): δppm 7.28-7.39 (m, 5 H), 5.09 (s, 2 H), 4.82 (br t, J=4.63 Hz, 1 H), 3.97 (br d, J=13.63 Hz, 1 H), 3.80 (br d, J=12.76 Hz, 2 H), 3.34-3.49 (m, 2 H), 2.68-3.07 (m, 4 H), 1.99 (s, 1 H), 1.40 (s, 9 H), 1.17 (t, J=7.07 Hz, 1 H) ppm. MS (ESI) m / z295.2 (M-tBu+H) +. Example 15B(R)-2-(methoxymethyl)piperazine-1,4-dicarboxylic acid 1-benzyl-4-tert-butyl ester

[0140] Example 15A (2.0 g, 5.71 mmol) was dissolved in dichloromethane (40 mL), and the mixture was cooled to 0°C. A fraction of 1,8-bis(dimethylamino)naphthalene (4.89 g, 22.83 mmol) was added to the mixture, followed by the slow addition of trimethoxyonium tetrafluoroborate (3.38 g, 22.83 mmol) over 5 minutes. The mixture was stirred at 0°C for 30 minutes, then heated to 20°C and stirred for 1 hour. The reaction mixture was extracted with ethyl acetate (five times). The combined organic layers were washed with brine, dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica column chromatography using ethyl acetate / petroleum ether to provide the title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d⁶): δppm 7.27–7.40 (m, 4H), 5.01–5.17 (m, 2H), 4.18 (br d, J=0.88 Hz, 1H), 3.68–3.97 (m, 3H), 3.32 (d, J=2.26 Hz, 2H), 3.24 (br s, 3H), 2.68–3.04 (m, 3H), 1.40 (s, 9H) ppm. Example 15C(R)-3-(methoxymethyl)piperazine-1-carboxylic acid tributyl ester

[0141] Example 15B (1.3 g, 3.57 mmol) was added to a suspension of Pd / C (10 wt%, 0.38 g, 0.357 mmol) in ethyl acetate (26 mL). The reaction mixture was hydrogenated using a hydrogen balloon at ambient temperature for 16 hours. The mixture was filtered through a diatomaceous earth pad, and the filtrate was concentrated under reduced pressure to provide the crude title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d6): δppm 3.65–3.87 (m, 2H), 3.26 (s, 3H), 3.20–3.22 (m, 1H), 3.22 (d, J=6.02 Hz, 1H), 2.83 (br d, J=11.80 Hz, 1H), 2.59–2.77 (m, 2H), 2.46–2.56 (m, 2H), 2.30 (br s, 1H), 1.40 (s, 9H) ppm. Example 15D(R)-3-(methoxymethyl)-4-(6-nitropyridin-3-yl)piperazine-1-carboxylic acid tributyl ester

[0142] At ambient temperature, Example 15C (800 mg, 3.47 mmol) and N,N-diisopropylethylamine (1.820 mL, 10.42 mmol) were added to a mixture of 5-fluoro-2-nitropyridine (740 mg, 5.21 mmol) in dimethyl sulfoxide (16 mL). The reaction mixture was heated to 120 °C and stirred for 12 hours. The reaction mixture was cooled and poured into water, and the mixture was extracted twice with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica column chromatography using ethyl acetate / petroleum ether to provide the title compound. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 8.64 (d, J=2.88 Hz, 1 H), 8.44 (d, J=8.88 Hz, 1 H), 8.22 (d, J=3.00 Hz, 1 H), 8.16 (d, J=9.26 Hz, 1 H), 8.04 (dd, J=8.94, 2.81 Hz, 1 H), 7.44 (dd, J=9.26, 3.00 Hz, 1 H), 4.34 (br s, 1 H), 3.96-4.07 (m, 1 H), 3.90 (br d, J=1.63 Hz, 1 H), 3.80 (br d, J=12.88 Hz, 1 H), 3.36 - 3.48 (m, 2 H), 3.32 (s, 1 H), 3.23 (s, 5 H), 1.42 (s, 9 H) ppm. MS (ESI) m / z 353.3 (M+H) +. Example 15E(R)-4-(6-aminopyridin-3-yl)-3-(methoxymethyl)piperazine-1-carboxylic acid tributyl ester

[0143] Example 15D (1.2 g, 3.41 mmol) was added to a suspension of Pd / C (10 wt%, 362 mg, 0.341 mmol) in ethyl acetate (36 mL). The reaction mixture was hydrogenated using a hydrogen balloon at ambient temperature for 16 hours. The reaction mixture was filtered through a diatomaceous earth pad, and the filtrate was concentrated under reduced pressure to provide the crude title compound. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 7.62 (d, J=2.63 Hz, 1 H), 7.21 (dd, J=8.76, 2.50 Hz, 1 H), 6.40 (d, J=8.76 Hz, 1 H), 5.51 (br s, 2 H), 3.39-3.69 (m, 3 H), 3.01-3.18 (m, 5 H), 2.72-2.94 (m, 2 H), 1.41 (s, 9 H) ppm. Example 15F (R)-4-(6-((6-(4-((R)-1-(3-(tributyl)-1,2,4-diazol-5-methamido)ethyl)-3-methylphenyl)pyrimidin-4-yl)amino)pyridin-3-yl)-3-(methoxymethyl)piperazine-1-carboxylic acid tributyl ester

[0144] To a solution of Example 1E (300 mg, 0.75 mmol) in 1,4-dimethylamine (5 mL), Example 15E (363 mg, 1.13 mmol), Pd₂(dba)₃ (CAS 52409-22-0, 69 mg, 0.075 mmol), XantPhos (CAS 161265-03-8, 43 mg, 0.075 mmol) and cesium carbonate (489 mg, 1.50 mmol) were added. The mixture was degassed using an argon stream and stirred at 100 °C for 12 hours. The reaction mixture was cooled, diluted with ethyl acetate, filtered through diatomaceous earth, concentrated under reduced pressure, and purified by silica chromatography using ethyl acetate and petroleum ether to give the title compound. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δ ppm 10.02 (s, 1 H), 9.91 (d, J=7.89 Hz, 1 H), 8.70 (s, 1 H), 8.02 (br d, J=2.85 Hz, 1 H), 7.79 - 7.87 (m, 2 H), 7.65 - 7.74 (m, 1 H), 7.60 (d, J=8.55 Hz, 1 H), 7.46 (dd, J=9.21, 2.63 Hz, 1 H), 7.25 - 7.35 (m, 1 H), 5.24 - 5.39 (m, 1 H), 3.69 - 4.00 (m, 3 H), 3.24 - 3.33 (m, 2 H), 3.20 (br s, 5 H), 2.89 - 3.10 (m, 2 H), 2.47 (s, 3 H), 1.51 (d, J=6.80 Hz, 3 H), 1.42 (s, 9 H), 1.36 (s, 9 H) ppm. LC / MS (ESI+): m / z 686.5 (M+H)+. Example 15G 3-(tributyl)-N-((R)-1-(4-(6-((5-(((R)-2-(methoxymethyl)piperazine-1-yl]pyridin-2-yl)amino)pyrimidin-4-yl]-2-methylphenyl)ethyl)-1,2,4-diazol-5-methylamine

[0145] At ambient temperature, the hydrochloride (4N contained in diane, 3 mL) was added dropwise to a solution of Example 15F (450 mg, 0.66 mmol) in 1,4-diane (3 mL). The mixture was stirred for 1 hour and concentrated under reduced pressure to give the crude title compound as hydrochloride. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δ ppm 9.97 (s, 1 H), 9.91 (br d, J=8.11 Hz, 1 H), 8.69 (s, 1 H), 7.95 - 8.00 (m, 1 H), 7.80 - 7.86 (m, 2 H), 7.55 - 7.70 (m, 2 H), 7.38 - 7.43 (m, 1 H), 7.27 - 7.34 (m, 1 H), 5.27 - 5.38 (m, 1 H), 3.67 - 3.81 (m, 2 H), 3.56 (s, 6 H), 3.51 - 3.61 (m, 1 H), 3.34 (s, 3 H), 3.14 - 3.20 (m, 3 H), 3.12 - 3.20 (m, 1 H), 2.92 - 3.05 (m, 1 H), 2.90 - 3.03 (m, 1 H), 2.78 - 2.86 (m, 1 H), 2.65 - 2.74 (m, 1 H), 2.47 (s, 3 H), 1.51 (br d, J=6.80 Hz, 3 H), 1.33 - 1.39 (m, 9 H) ppm. LC / MS (ESI+): m / z586.4 (M+H) +. Example 15H 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2R)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-(methoxymethyl)piperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-methylphenyl}ethyl]-1,2,4-diazol-5-methylamine

[0146] Example 15G (130 mg, 0.22 mmol), Example 1J (115 mg, 0.33 mmol), N,N-diisopropylethylamine (194 µL, 1.11 mmol), and sodium iodide (100 mg, 0.66 mmol) were mixed in acetonitrile (5 mL). The mixture was stirred at 80 °C for 16 hours. The mixture was cooled, quenched with a few drops of formic acid, and directly subjected to reversed-phase preparative HPLC to isolate the title compound (Phenomenex Luna® C18 column, 30 × 75 mm × 3 µm, 0.1% formic acid / water and acetonitrile as mobile phase). 1H NMR (400 MHz, dimethyl sulfoxide-d 6): δppm 10.33 (s, 1 H), 9.96 (s, 1 H), 9.88 (br d, J=7.65 Hz, 1 H), 8.70 (s, 1 H), 8.14 (s, 1 H), 8.00 (br d, J=2.38 Hz, 2 H), 7.76-7.91 (m, 2 H), 7.64-7.73 (m, 1 H), 7.60 (br d, J=8.66 Hz, 1 H), 7.44 (br dd, J=8.91, 2.64 Hz, 1 H), 7.18-7.31 (m, 4 H), 5.33 (quin, J=7.09 Hz, 1 H), 3.85-3.96 (m, 1 H), 3.76 (t, J=6.65 Hz, 2 H), 3.51-3.68 (m, 1 H), 3.29 (br s, 1 H), 3.17 (s, 3 H), 2.85-3.08 (m, 3 H), 2.73-2.82 (m, 2 H), 2.69 (br t, J=6.65 Hz, 2 H), 2.51-2.63 (m, 3 H), 2.47 (s, 3 H), 2.17-2.34 (m, 2 H), 1.51 (br d, J=6.90 Hz, 3 H), 1.36 (s, 9 H) ppm. MS (ESI) m / z802.3 (M+H) +.Example 16 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-(trifluoromethyl)phenyl}ethyl]-1,2,4-diazol-5-methamide Example 16A (S, E)-N-(4-bromo-2-(trifluoromethyl)benzylidene)-2-methylpropane-2-sulfinamide.

[0147] 4-Bromo-2-(trifluoromethyl)benzaldehyde (10.00 g, 39.5 mmol) and dichloromethane (100 mL) were added to a 500 mL flask. At ambient temperature, (S)-2-methylpropane-2-sulfinamide (6.23 g, 51.4 mmol) was added to the stirred mixture, followed by titanium ethoxide (IV) (18.03 g, 79.0 mmol). The mixture was stirred at 30 °C for 12 hours and quenched with ethyl acetate (50 mL) and water (50 mL). The mixture was filtered, and the organic phase was separated. The aqueous phase was extracted with ethyl acetate (100 mL, three times), and the combined organic phases were dried over sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give the crude title compound. ¹H NMR (400 MHz, CDCl₃): δppm 8.89 (d, J=1.5 Hz, ¹H), 8.09 (d, J=8.4 Hz, ¹H), 7.91 (d, J=1.3 Hz, ¹H), 7.79 (d, J=8.4 Hz, ¹H), 1.31–1.21 (m, 9H) ppm. Example 16B(S)-N-((R)-1-(4-bromo-2-(trifluoromethyl)phenyl)ethyl)-2-methylpropane-2-sulfinamide

[0148] Anhydrous toluene (60 mL) and Example 16A (5.40 g, 15.16 mmol) were added to a 250 mL flask. The mixture was cooled to -5 °C. CH3MgBr (3.0 M in tetrahydrofuran, 10.1 L, 30.3 mmol) was added dropwise. The mixture was then stirred at 0 °C for 3 hours, and a saturated aqueous solution of NH4Cl (120 mL) was added to quench the reaction. The organic phase was separated, and the aqueous phase was extracted with ethyl acetate (30 mL, three times). The combined organic layers were washed with brine (30 mL), dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica column chromatography using ethyl acetate and petroleum ether to give the title compound. 1H NMR (400 MHz, dimethyl sulfoxide): δppm 7.92 (dd, J=8.50, 1.75 Hz, 1 H), 7.81 (d, J=1.88 Hz, 1 H), 7.74 (d, J=8.50 Hz, 1 H), 5.63 (d, J=4.50 Hz, 1 H), 4.69 (quin, J=5.78 Hz, 1 H), 1.44 (d, J=6.63 Hz, 3 H), 1.06 (s, 9 H) ppm. MS (ESI) m / z372.0, 374.0 (M+H) +. Example 16C(S)-N-((R)-1-(2-trifluoromethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethyl)-2-methylpropane-2-sulfinamide

[0149] At 20°C, bis(pinacol)diboron (853 mg, 3.36 mmol) and potassium acetate (527 mg, 5.37 mmol) were added to a solution of Example 16B (1.00 g, 2.69 mmol) in 20 mL of 1,4-dioxane. The reaction vessel was evacuated and backfilled with nitrogen three times, followed by the addition of a 1,1'-bis(diphenylphosphino)ferrocene-dichloropalladium(II)dichloromethane complex (197 mg, 0.27 mmol). The reaction vessel was again evacuated and backfilled with nitrogen three times. The reaction mixture was heated to 80°C and stirred for 16 hours. The reaction mixture was cooled and filtered through a diatomaceous earth filter. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography, eluting with petroleum ether and ethyl acetate to provide the title compound. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 7.93 (br d, J=6.58 Hz, 1 H), 7.71 - 7.89 (m, 2 H), 5.62 (br d, J=3.73 Hz, 1 H), 4.66 - 4.82 (m, 1 H), 1.45 (br d, J=6.14 Hz, 3 H), 1.30 (s, 9 H), 1.06 (br d, J=2.19 Hz, 12 H) ppm. MS (ESI) m / z420.2 (M+H) +. Example 16D(R)-1-(2-trifluoromethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethane-1-amine

[0150] At ambient temperature, the hydrochloride salt (4N contained in 1,4-dimethylamine, 10 mL) was added dropwise to a solution of Example 16C (900 mg, 2.15 mmol) in 1,4-dimethylamine (5 mL). The mixture was stirred for 1 hour and then concentrated under reduced pressure to give the crude title compound as hydrochloride. ¹H NMR (400 MHz, dimethyl sulfoxide-d6): 8.94 (br s, 3 H), 8.01–8.15 (m, 2 H), 7.93 (s, 1 H), 4.52 (br d, J=4.75 Hz, 1 H), 1.54 (d, J=6.63 Hz, 3 H), 1.31 (s, 12 H) ppm. MS (ESI) m / z 316.2 (M+H)+. Example 16E (R)-3-(tert-butyl)-N-(1-(2-trifluoromethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethyl)-1,2,4-diazol-5-methylamine

[0151] At 20°C, triethylamine (622 mg, 6.14 mmol) was added to a solution of Example 16D (360 mg, 1.02 mmol) and methyl 3-(tributyl)-1,2,4-diazol-5-carboxylate (471 mg, 2.56 mmol) in ethanol (8 mL). The mixture was stirred at 60°C for 16 hours, followed by concentration under reduced pressure to remove ethanol. The crude material was purified by silicone column chromatography, eluted with petroleum ether and ethyl acetate to provide the title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d6): δppm 10.04 (d, J=7.23 Hz, 1 H), 7.93–8.00 (m, 1 H), 7.84–7.92 (m, 2 H), 5.40 (quin, J=6.91 Hz, 1 H), 1.49 (d, J=6.80 Hz, 3 H), 1.36 (s, 9 H), 1.30 (s, 12 H) ppm. MS (ESI) m / z 468.2 (M+H)+. Example 16F(R)-3-(tributyl)-N-(1-(4-(6-chloropyrimidin-4-yl)-2-(trifluoromethyl)phenyl)ethyl)-1,2,4-diazol-5-methylamine

[0152] A solution of potassium phosphate (209 mg, 0.98 mmol) in water (2.5 mL) and [1,1'-bis(diphenylphosphine)ferrocene]dichloropalladium(II) (22 mg, 0.03 mmol) were added to a mixture of Example 16E (230 mg, 0.49 mmol) and 4,6-dichloropyrimidine (147 mg, 0.98 mmol) in tetrahydrofuran (10 mL). The mixture was degassed with a stream of nitrogen at ambient temperature and stirred at 80 °C for 2 hours under a nitrogen atmosphere. The mixture was cooled, poured into water, and extracted with ethyl acetate (three times). The combined organic layers were washed with brine, dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silicon column chromatography, eluted with ethyl acetate / hexane, to give the crude title compound. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 10.10 (br d, J=7.23 Hz, 1 H), 9.14 (s, 1 H), 8.52 - 8.59 (m, 2 H), 8.49 (s, 1 H), 8.06 (d, J=8.11 Hz, 1 H), 5.45 (br t, J=6.91 Hz, 1 H), 1.55 (d, J=6.80 Hz, 3 H), 1.37 (d, J=1.32 Hz, 9 H) ppm. MS (ESI) m / z454.1 (M+H) +. Example 16G (S)-4-(6-((6-(4-((R)-1-(3-(tributyl)-1,2,4-diazol-5-methamido)ethyl)-3-(trifluoromethyl)phenyl)pyrimidin-4-yl)amino)pyridin-3-yl)-3-methylpiperazine-1-carboxylic acid tributyl ester

[0153] To a solution of Example 16F (170 mg, 0.38 mmol) in 1,4-dimethylamine (4 mL), Example 1L (138 mg, 0.45 mmol), Pd₂(dba)₃ (CAS 52409-22-0, 21 mg, 0.022 mmol), XantPhos (CAS 161265-03-8, 22 mg, 0.037 mmol), and cesium carbonate (244 mg, 0.75 mmol) were added. The mixture was degassed using a nitrogen stream and stirred at 100 °C for 16 hours under a nitrogen atmosphere. The reaction mixture was cooled, diluted with ethyl acetate, and filtered through diatomaceous earth. The filtrate was concentrated under reduced pressure, and the residue was purified by silica chromatography using ethyl acetate and petroleum ether to give the title compound. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 10.24 (br s, 1 H), 10.10 (d, J=7.23 Hz, 1 H), 8.78 (s, 1 H), 8.34 (s, 1 H), 8.29 (br d, J=8.33 Hz, 1 H), 7.99 - 8.07 (m, 2 H), 7.67 - 7.78 (m, 1 H), 7.49 (br dd, J=8.99, 2.41 Hz, 1 H), 7.25 - 7.35 (m, 1 H), 5.45 (br t, J=7.13 Hz, 1 H), 3.77 - 3.92 (m, 2 H), 3.56 - 3.69 (m, 1 H), 3.26 - 3.37 (m, 1 H), 3.19 (br d, J=11.18 Hz, 2 H), 2.91 - 3.01 (m, 1 H), 1.56 (br d, J=6.80 Hz, 3 H), 1.42 (s, 9 H), 1.37 (s, 9 H), 0.89 (d, J=6.58 Hz, 3 H) ppm. LC / MS (ESI+): m / z710.4 (M+H) +. Example 16H 3-(tert-butyl)-N-((R)-1-(2-trifluoromethyl-4-(6-((5-((S)-2-methylpiperazine-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)phenyl)ethyl)-1,2,4-diazol-5-methylamine

[0154] The hydrochloride salt (4 N contained in methanol, 20 mL) was added dropwise to a solution of Example 16 G (100 mg, 0.14 mmol) in methanol (20 mL). The reaction mixture was stirred at ambient temperature for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude title compound as the hydrochloride salt. 1H NMR (400 MHz, dimethyl sulfoxide - d 6): δppm 9.99 - 10.17 (m, 2 H), 8.75 (s, 1 H), 8.35 (s, 1 H), 8.28 (br d, J=8.25 Hz, 1 H), 7.93 - 8.18 (m, 3 H), 7.65 - 7.78 (m, 1 H), 7.41 (dd, J=9.13, 2.75 Hz, 1 H), 7.22 - 7.35 (m, 1 H), 5.45 (quin, J=6.69 Hz, 1 H), 3.68 - 3.78 (m, 1 H), 3.18 - 3.35 (m, 1 H), 3.01 - 3.12 (m, 1 H), 2.81 - 2.98 (m, 3 H), 2.66 - 2.77 (m, 2 H), 1.55 (br d, J=6.88 Hz, 3 H), 1.37 (s, 9 H), 0.96 (d, J=6.38 Hz, 3 H) ppm. LC / MS (ESI+): m / z610.3 (M+H) +. Example 16I 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-(trifluoromethyl)phenyl}ethyl]-1,2,4-diazol-5-methylamine

[0155] To a solution of Example 16H (63 mg, 0.11 mmol) in acetonitrile (2 mL), add Example 1J (56 mg, 0.16 mmol), sodium iodide (65 mg, 0.43 mmol), and N,N-diisopropylethylamine (56 µL, 0.32 mmol). Stir the mixture at 80 °C for 16 hours. Cool the mixture to ambient temperature, filter, and directly subject it to reversed-phase preparative HPLC to isolate the title compound (Waters™ Xbridge C18 column, 30 × 100 mm × 10 µm, 10 mM ammonium bicarbonate / water and acetonitrile as mobile phase). 1H NMR (400 MHz, dimethyl sulfoxide-d 6): δppm 10.34 (br s, 1 H), 10.10 (s, 2 H), 8.75 (s, 1 H), 8.35 (s, 1 H), 8.28 (br d, J=8.25 Hz, 1 H), 8.10 (br s, 1 H), 8.02 (br d, J=10.38 Hz, 2 H), 7.72 (br s, 1 H), 7.39 - 7.54 (m, 1 H), 7.17 - 7.33 (m, 4 H), 5.45 (br d, J=5.88 Hz, 1 H), 3.88 (br d, J=5.13 Hz, 1 H), 3.76 (br t, 2.66 - 2.73 (m, 4 H), 2.44 (br d, J=11.63 Hz, 2 H), 2.31 (br d, J=11.63 Hz, 1 H), 1.56 (br d, J=6.75 Hz, 3 H), 1.37 (s, 9 H), 0.99 (br d, J=6.25 Hz, 3 H) ppm. MS (ESI) m / z826.4 (M+H) +.Example 17 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-fluorophenyl}ethyl]-1,2,4-diazol-5-methylamine Example 17A (S, E)-N-(4-bromo-2-fluorobenzyl)-2-methylpropane-2-sulfinamide.

[0156] Triisopropyl borate (243 g, 1.29 mol), 4-bromo-2-fluorobenzaldehyde (250 g, 1.23 mol), and (S)-2-methylpropane-2-sulfinamide (179 g, 1.48 mol) were added to a 1 L reactor at ambient temperature. The mixture was stirred at 70 °C for 12 hours. Water (500 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (500 mL × 3). The combined organic phases were washed with brine (1.0 L), dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to provide the title compound. ¹H NMR (400 MHz, CDCl₃): δppm 8.82 (s, ¹H), 7.81–8.92 (m, ¹H), 7.30–7.47 (m, 2H), 1.26 (s, 9H) ppm. Example 17B (S)-N-((R)-1-(4-bromo-2-fluorophenyl)ethyl)-2-methylpropane-2-sulfinamide

[0157] Anhydrous dichloromethane (3.0 L) and Example 16A (150 g, 0.49 mol) were added to a 5 L reactor. The mixture was cooled to 0 °C. CH3MgBr (3.0 M contained in tetrahydrofuran, 245 L, 0.73 mmol) was added dropwise. The mixture was heated to ambient temperature and stirred for 12 hours, followed by the addition of a saturated aqueous solution of NH4Cl (1.0 L) to quench the reaction. The organic phase was separated, and the aqueous phase was extracted with dichloromethane (1.0 L, three times). The combined organic layers were washed with brine (2.0 L), dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give the crude title compound. ¹H NMR (400 MHz, CDCl₃): δppm 7.21–7.28 (m, 3H), 4.73–4.90 (m, 1H), 3.33 (d, J=4.03 Hz, 1H), 1.56 (d, J=6.85 Hz, 3H), 1.20 (s, 9H) ppm. Example 17C(R)-1-(4-bromo-2-fluorophenyl)ethane-1-amine

[0158] Example 17B (87.0 g, 0.27 mol) and methanol (870 mL) were added to a 1 L reactor. The hydrochloride salt (4 N in methanol, 435 mL, 1.74 mol) was added to the stirred solution at 0 °C. The reaction mixture was stirred at 0 °C for 2 hours and concentrated under reduced pressure to give the crude title compound as its hydrochloride salt. ¹H NMR (400 MHz, dimethyl sulfoxide-d6): δppm 8.80 (s, 3H), 7.59–7.76 (m, 2H), 7.53 (dd, J=8.38, 1.41 Hz, 1H), 4.41–4.63 (m, 1H), 1.44–1.57 (m, 3H) ppm. Example 17D(R)-(1-(4-bromo-2-fluorophenyl)ethyl)aminocarbamate tributyl ester

[0159] Example 17C (65.0 g, 0.30 mol), tetrahydrofuran (650 mL), and water (650 mL) were added to a 3 L reactor. Sodium hydroxide (35.8 g, 0.89 mol) and di-tert-butyl dicarbonate (78.1 g, 0.36 mol) were added to the stirred solution at 0 °C. The reaction was heated to ambient temperature and stirred for 2 hours. The mixture was diluted with water (1.0 L) and extracted with ethyl acetate (1.0 L, three times). The combined organic layers were washed with brine (2.0 L), dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica column chromatography with ethyl acetate and petroleum to give the title compound. ¹H NMR (400 MHz, CD 3OD): δppm 7.18–7.39 (m, 3H), 4.88–4.95 (m, 1H), 1.28–1.49 (m, 12H) ppm. Example 17E(R)-(1-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethyl)aminocarbamate tributyl ester

[0160] At 20°C, bis(pinacolyl)diboron (72.8 g, 0.29 mol) and potassium acetate (46.9 g, 0.48 mol) were added to a solution of Example 17D (76.0 g, 0.24 mol) in 1,4-dimethylbenzene (760 mL). The reaction vessel was evacuated and backfilled with nitrogen three times, followed by the addition of a 1,1'-bis(diphenylphosphino)ferrocene-dichloropalladium(II)dichloromethane complex (17.5 g, 0.02 mol). The reaction vessel was again evacuated and backfilled with nitrogen three times. The reaction mixture was heated to 100°C and stirred under a nitrogen atmosphere for 16 hours. The reaction mixture was cooled and filtered through a diatomaceous earth pad. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography, eluted with petroleum ether and ethyl acetate to provide the title compound. ¹H NMR (400 MHz, CDCl₃): δppm 7.54 (d, J = 7.50 Hz, ¹H), 7.45 (d, J = 11.03 Hz, ¹H), 7.28–7.32 (m, ¹H), 4.99 (s, 2H), 1.42 (s, 12H), 1.34 (s, 12H) ppm. Example 17F(R)-1-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethane-1-amine

[0161] At ambient temperature, the hydrochloride salt (4N contained in 1,4-dimethylamine, 14 mL) was added dropwise to a solution of Example 17E (1030 mg, 2.15 mmol) in dichloromethane (20 mL). The mixture was stirred for 3 hours and then concentrated under reduced pressure to give the crude title compound as hydrochloride. ¹H NMR (400 MHz, dimethyl sulfoxide-d6): 8.68 (br s, 3H), 7.64–7.72 (m, 1H), 7.56 (d, J=7.58 Hz, 1H), 7.40 (d, J=10.64, 1H), 4.62 (br s, 1H), 1.52 (d, J=6.85 Hz, 3H), 1.31 (s, 12H) ppm. MS (ESI) m / z 249.2 (M+H) +. Example 17G (R)-3-(tributyl)-N-(1-(2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)phenyl)ethyl)-1,2,4-diazol-5-methylamine

[0162] At ambient temperature, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU, 1.28 mL, 8.46 mmol) was added to a solution of Example 17F (850 mg, 2.82 mmol) and methyl 3-(tributyl)-1,2,4-diazol-5-carboxylate (779 mg, 4.23 mmol) in 10 mL of 2-methyltetrahydrofuran. The mixture was stirred for 2 hours and then diluted with ethyl acetate (50 mL) and brine (20 mL). The organic phase was separated and the aqueous phase was extracted with ethyl acetate (20 mL). The combined organic phases were dried over sodium sulfate and filtered. The filtrate was concentrated under reduced pressure and eluted with hexane and ethyl acetate by silica gel column chromatography to provide the title compound. ¹H NMR (400 MHz, dimethyl sulfoxide-d⁶): 9.95 (d, J=7.70 Hz, 1H), 7.49–7.63 (m, 2H), 7.39 (d, J=10.64 Hz, 1H), 5.42 (t, J=7.27 Hz, 1H), 1.56 (d, J=6.97 Hz, 3H), 1.42 (s, 9H), 1.35 (s, 12H) ppm. MS (ESI) m / z 418.2 (M+H)⁺. Example 17H(R)-3-(tributyl)-N-(1-(4-(6-chloropyrimidin-4-yl)-2-fluorophenyl)ethyl)-1,2,4-acediazole-5-methylamine

[0163] A solution of potassium phosphate (775 mg, 3.65 mmol) in water (2 mL) and [1,1'-bis(diphenylphosphine)ferrocene]dichloropalladium(II) (149 mg, 0.18 mmol) were added to a mixture of Example 17G (762 mg, 1.83 mmol) and 4,6-dichloropyrimidine (326 mg, 2.19 mmol) in 1,4-dimethylamine (7.3 mL). The mixture was degassed at ambient temperature using a nitrogen stream and stirred at 85 °C for 20 min under a nitrogen atmosphere. The mixture was cooled, poured into water, and extracted three times with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silicon column chromatography, eluting with ethyl acetate / hexane to give the crude title compound. MS (ESI) m / z 404.1 (M+H)+. Example 17I (S)-4-(6-((6-(4-((R)-1-(3-(tributyl)-1,2,4-diazol-5-methamido)ethyl)-3-fluorophenyl)pyrimidin-4-yl)amino)pyridin-3-yl)-3-methylpiperazine-1-carboxylic acid tributyl ester

[0164] To a solution of Example 17G (268 mg, 0.66 mmol) in 1,4-dimethylamine (10 mL), Example 1L (349 mg, 1.20 mmol), racemic BINAP-Pd-G3 (CAS 2151915-22-7, 66 mg, 0.066 mmol), and cesium carbonate (649 mg, 1.99 mmol) were added. The mixture was heated at 75 °C under a nitrogen atmosphere for 40 minutes. The mixture was cooled, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica chromatography with ethyl acetate and hexane to provide the title compound. LC / MS (ESI+): m / z 660.4 (M+H)+. Example 17J 3-(tert-butyl)-N-((R)-1-(2-fluoro-4-(6-((5-((S)-2-methylpiperazine-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)phenyl)ethyl)-1,2,4-diazol-5-methylamine

[0165] To a solution of Example 17I (346 mg, 0.52 mmol) in dichloromethane (10 mL), hydrochloride (4 N contained in 1,4-dimethylamine, 6.6 mL) was added. The reaction mixture was stirred at ambient temperature for 2 hours. The reaction mixture was concentrated under reduced pressure and purified by silica column chromatography using dichloromethane and methanol (with 7 N ammonia / methanol added) to give the title compound. LC / MS (ESI+): m / z 560.3 (M+H)+. Example 17K 3-Tributyl-N-[(1R)-1-{4-[6-({5-[(2S)-4-{2-[4-(2,4-dioxy-1,3-diazacyclohexane-1-yl)phenyl]ethyl}-2-methylpiperazine-1-yl]pyridin-2-yl}amino)pyrimidin-4-yl]-2-fluorophenyl}ethyl]-1,2,4-diazol-5-methylamine

[0166] To a solution of Example 17J (80 mg, 0.14 mmol) in N,N-dimethylformamide (1 mL), Example 1J (56 mg, 0.16 mmol), potassium iodide (119 mg, 0.72 mmol), and N,N-diisopropylethylamine (50 µL, 0.29 mmol) were added. The mixture was stirred at 80 °C for 2 hours. After cooling to ambient temperature, it was filtered, and the filtrate was directly subjected to reversed-phase preparative HPLC to isolate the title compound (Phenomenex Gemini® NX C18 column, 30 × 250 mm × 5 µm, 0.1% ammonium hydroxide / water and acetonitrile as mobile phase). 1H NMR (500 MHz, dimethyl sulfoxide - d 6): δppm 10.30 (s, 1H), 9.98 (s, 1H), 9.86 (d, J = 7.8 Hz, 1H), 8.66 (s, 1H), 8.00 (s, 1H), 7.94 (d, J = 2.9 Hz, 1H), 7.78 (dd, J = 8.1, 1.7 Hz, 1H), 7.71 (dd, J = 11.5, 1.7 Hz, 1H), 7.66 – 7.57 (m, 2H), 7.37 (dd, J = 9.3, 2.9 Hz, 1H), 7.21 (d, J = 8.4 Hz, 2H), 7.17 (d, J = 8.4 Hz, 2H), 5.34 (p, J = 7.2 Hz, 1H), 3.80 (s, 1H), 3.69 (t, J = 6.7 Hz, 2H), 3.15 (d, J = 11.3 Hz, 1H), 2.93 (dd, J = 10.6 Hz, 1H), 2.79 (d, J = 10.4 Hz, 1H), 2.71 (dd, J = 7.7, 7.7 Hz, 2H), 2.66 – 2.59 (m, 2H), 2.55 – 2.42 (m, 2H), 2.38 (d, J = 10.9 Hz, 1H), 2.31 – 2.20 (m, 2H), 1.49 (d, J = 7.0 Hz, 3H), 1.30 (s, 9H), 0.93 (d, J = 6.3 Hz, 3H) ppm. MS (ESI) m / z 776.4 (M+H)+. Overview of BTK screening.

[0167] Btk kinase activity was measured in vitro using a mobility shift assay (MSA). The ability of Btk phosphorylated fluorescent peptide acceptor (FAM-GEEPLYWSFPAKKK-NH2) was measured. Kinase reactions were assembled in 384-well disks with a total volume of 25 µL per well. The following were added to each well: compound buffer (or control); enzyme buffer; and acceptor buffer, as further described below.

[0168] Specifically, the following were added: (1) Compound buffer or control: 5 µl 5X compound buffer [(5X compound buffer contains: 1X master buffer containing X µM of the test compound in 5% dimethyl sulfoxide; (2X master buffer containing 200 mM HEPES, pH 7.5, 0.2% BSA, and 0.02% Triton X-100)]; and (2) Enzyme buffer: 10 µL 2.5X enzyme buffer (1X master buffer, 12.5 mM MgCl2, 2.5 mM DTT, 25 µM sodium orthovanadate, 25 µM β-glycerophosphate and 1.25 nM BTK enzyme). Human BTK enzyme Nanosyn-293HEK, wild type, is available from Nanosyn, Santa Clara, CA. The enzyme and compound were pre-incubated for 15 minutes. In addition, the following items were added: (3) Acceptance buffer: 10 µL 2.5X acceptance buffer (1X master buffer, 50 µM ATP and 2.5 µM peptide acceptor FAM). Each plate was incubated at 25°C for 3 hours. The reaction was terminated by adding 45 µL 1.55X stop buffer (1X master buffer and 31 mM EDTA) to each well. The final reaction mixture was as follows: 100 mM HEPES, pH 7.5; 0.1% BSA; 0.01% Triton X-100; 1 mM DTT; 5 mM MgCl2; 10 µM sodium orthovanadate; 10 µM β-glycerophosphate; 50 µM ATP; 1% dimethyl sulfoxide (from the compound); 1 µM fluorescent peptide acceptor and 0.5 nM BTK enzyme.

[0169] The terminated reaction was analyzed using a 12-channel LABCHIP® 3000 microfluidic instrument (available from Caliper Life Sciences, Waltham, MA). Enzymatic phosphorylation of the peptide leads to a change in net charge, allowing the product and acceptor peptides to be separated by electrophoresis. Two fluorescence peaks were observed when the acceptor and product peptides separated. The change in the relative fluorescence intensity of the acceptor and product peaks is a measured parameter reflecting enzyme activity. In the presence of active compounds, the ratio between product and acceptor is altered. The product signal weakens, while the acceptor signal strengthens. The capillary electrophoresis pattern (RDA acquisition file) was analyzed using HTS Well Analyzer software (available from Caliper Life Sciences, Waltham, MA).

[0170] The activity in each sample was determined as a product-to-sum ratio (PSR): P / (S+P), where P represents the peak height of the product peptide and S represents the peak height of the receptor peptide. For each compound, enzyme activity was measured at different concentrations (12 concentrations of compounds, separated by a 3x dilution interval). Four replicates were formed by negative control samples (0% inhibition in the absence of active compounds) and positive control samples (100% inhibition in the presence of 20 mM EDTA) and used to calculate the % inhibition value of each active compound at each concentration. The percentage inhibition (Pinh) was determined using the following equation: Pinh = (PSR0% - PSRinh) / (PSR0% - PSR100%)*100, where PSRinh is the product-to-sum ratio in the presence of inhibitors, PSR0% is the average product-to-sum ratio in the absence of active compounds, and PSR100% is the average product-to-sum ratio in the 100% inhibition control samples.

[0171] The IC50 value of the active compound was determined by fitting a 4-parameter sigmoid dose-response model to the inhibition curve (Pinh versus inhibitor concentration) using XLfit® 4 software (IDBS, Boston, MA). The IC50 values ​​are reported in the BTK IC50 (nM) column of Table 2. Cerebellar binding analysis

[0172] Cerebellar binding was measured in vitro using the AlphaLISA® Human Cerebellum Binding Kit (PerkinElmer, Waltham, MA) according to the manufacturer's protocol ("Alpha" refers to amplified luminescent proximity homogeneous analysis). In this analysis, donor beads and acceptor beads approach each other via ligands that bind to the CRBN protein. Excitation of the donor bead triggers a single-peak release of oxygen, which in turn triggers a cascade of energy transfer reactions to the acceptor bead, resulting in a sharp peak of light emission at 615 nm.

[0173] Specifically, nickel chelate acceptor beads (PerkinElmer AL108 M) were used to capture 6XHIS-tagged CRBN protein, and streptavidin-coated donor beads (PerkinElmer 6760002) were used to capture 50 nM biotinylated ligands. Each well included the following: sample or control; HIS-tagged CRBN protein; biotinylated ligand; nickel chelate acceptor beads; and streptavidin donor beads.

[0174] Competitive Analysis: Using a Labcyte Echo acoustic dispenser, the test compound in 100% dimethyl sulfoxide was plated onto a white low-volume 384-well plate (PerkinElmer Proxiplate 384 plus # 6008289) with an 11-point dose profile and a 3-fold dilution protocol. After adding 10 µL of the His-tagged cerebellar / DDB 1 complex and 50 µM of the biotin-labeled ligand to the buffer (buffer: 10 mM HEPES pH 7.4, 150 mM NaCl, 0.05% Tween-20, and 10 mM DTT), this resulted in a maximum dose (initial concentration) of 10 µM and a final concentration of 0.1% DMSO. The plate was incubated at room temperature for 30 minutes.

[0175] Detection: Under low light conditions, 10 µL of detection reagent containing nickel chelate acceptor beads and streptavidin donor beads (Perkin Elmer 6760002) was added to achieve a final concentration of 20 g / ml. The disc was incubated for 1 hour and read on a PerkinElmer™ Envision™ disc reader (680 nm excitation and 570 nM emission settings). The IC50 of each compound was determined by calculating the percentage of inhibition without drug and protein control, and the Ki of each compound was calculated using Cheng-Prusoff conversion. The IC50 value was determined based on Ki. The IC50 values ​​are reported in the CRBN IC50 (nM) column of Table 2. Analysis of Bik protein MSD in human whole blood using ELISA.

[0176] Btk protein degradation was tested using the Meso Scale Discovery-ELISA system (Meso Scale Discovery-ELISA, MSD-ELISA): (1) the test compound was added to human whole blood; (2) human whole blood cells treated with the test compound were lysed; (3) the lysed cells were treated with a Btk antibody capture antibody; (4) the lysed cells were treated with an anti-mouse Btk primary antibody; and (5) the lysed cells were treated with an anti-mouse antibody (tag).

[0177] Specifically, approximately 80 µl of fresh human whole blood from a healthy donor was transferred to each well of a 96-well V-shaped dish (Costar, catalog number 3894). Using a TECAN™ D300e digital dispenser (Tecan), the compound was repeatedly added at a 1:4 serial dilution [(8 pt analysis)] [starting from a maximum concentration of 3 µM in DMSO solution (final 0.2%)]. The dish was then placed in an incubator at 37°C and incubated for 6 hours.

[0178] At the end of the incubation, add 80 µl of 2X CST dissolution buffer (Cell Signaling Technology, catalog number 9803) prepared from 10X stock solution to each well to dissolve the blood. Then seal the tray with a tray sealer and freeze the samples at -80°C overnight.

[0179] The total amount of BTK protein remaining in blood samples after compound treatment was measured using an MSD-ELISA. After thawing the frozen trays, the blood lysate was thoroughly mixed and transferred to a small-spot high-binding MSD tray (Meso Scale Discovery, catalog number L45XB-3), and coated with the capture antibody rabbit anti-human BTK (D3H5) (Cell Signaling Technology, catalog number 8547CF). Subsequently, the primary detection antibody mouse anti-human BTK monoclonal antibody (BD Biosciences, catalog number 611117) and the secondary detection antibody anti-mouse sulfonyl tag (Meso Scale Discovery, catalog number R32AC-1) were added. The analysis tray was read on an MSD S600 tray reader by adding 2X MSD read buffer (Meso Scale Discovery, R92TC-1). All reagent transfers, including those involving hemolysis, were performed using a Biomed i7 liquid processor. The DC 50 value was calculated by averaging data from repeated experiments based on the dose-response curves, and curve fitting was performed using GraphPad Prism with nonlinear regression. The DC 50 values ​​are reported in the hWB BTK DC 50 (nM) column of Table 2. The D max value represents the maximum amount of BTK degradation observed during compound treatment compared to samples treated with DMSO alone. The dMax (%) value is reported in the hWB BTK dMax (%) column of Table 2. Mouse bone marrow analysis.

[0180] This study used female C57BL / 6J mice (Jackson Laboratories). Compounds or mediators / controls were prepared using EtOH; PEG-400; Phosal 53 MCT (10:30:60, v / v). Four mice were tested for each compound or mediator / control. On day 1, mice were administered a single dose of dose A. On day 2 (24 hours later), tissue (bone marrow) was collected from one hind leg of each mouse.

[0181] Analysis based on MSD (Meso Scale Discovery) ELISA (enzyme-linked immunosorbent assay) was used to determine BTK degradation in mouse bone marrow. The following materials and equipment were used: 10% NP40 solution (nonionic polyoxyethylene surfactant, ThermoFisher, catalog number 28324); protease inhibitor mini tablets (Roche, catalog number 05 892 970 001); PhosphoSTOP™ tablets (Roche, catalog number 04 906 837 001); Pierce™ BCA protein analysis kit (ThermoFisher, catalog number 23225); Precellys® 2 mL dissolving tubes with 2.8 mm beads (VWR, catalog number 10032-756); Bertin Precellys® 24 / Cryolys® cooling system; 96-well MSD streptavidin pre-coated trays (MSD, catalog number L15SA-1); MSD inhibitor A (MSD, catalog number R93BA-4); capture antibody, biotinylated BTK mAb (BD). Biosciences, catalog number 624008; custom orders); primary detection antibody, BTK RpAb (rabbit multi-strain antibody, CST, catalog number 8547S, D3H5 strain); secondary detection antibody, anti-rabbit sulfonyl tag (MSD, catalog number R32AB-1); read buffer, 4X (MSD, catalog number R92TC-1); recombinant BTK protein (Thermofisher, catalog number PR5442A); MSD Tris wash buffer (10x) (MSD, catalog number R61TX-1); PBS (phosphate-buffered saline); deionized water (diH2O); dissolution buffer [PBS in 0.2% NP40+ protease inhibitor tablets (1 tablet / 10 mL) + PhosphoSTOP tablets (1 tablet / 10 mL)]; 3% MSD blocker A [MSD blocker A + 450 mL diH2O + 50 mL]; and 10x MSD Tris wash buffer. Sample processing for programmed PD analysis: Preparation of mouse bone marrow lysate

[0182] Prepare lysis buffer [(PBS + 0.2% NP40 + PI (protease inhibitor) + PhosphoSTOP)] and place it on ice until completely cooled. Add ice-cold lysis buffer (300 µl) to each Precellys® tube containing bead and bone marrow samples. Transfer the sample tubes to a Precellys® instrument and homogenize using 5000 rpm - 2 × 20 seconds. After homogenization, centrifuge the sample lysate at 14,000 rpm for 20 minutes at 4°C. Protein estimation (BCA)

[0183] Following the manufacturer's recommended procedure, use the Pierce™ BCA protein analysis kit to determine protein concentration. Prepare protein standardization working solutions, using 30 µg of protein per sample solution for analysis. Standard curve sample preparation.

[0184] Prepare a 300 ng / ml working stock solution of recombinant BTK protein in dissolution buffer, and serially dilute the sample 3-fold (10-point analysis, with the highest concentration being 300 ng / ml and the lowest concentration being 0.015 ng / ml) for MSD analysis on day 1.

[0185] MSD Disk Coating: Prepare a stock solution of coated antibody (biotinylated BTK mAb) at 1 µg / mL in PBS buffer. Add 50 µL / well of the capture antibody (biotinylated BTK mAb) solution and gently agitate the disk for two hours at ambient temperature. Wash the disk three times with 1X MSD Tris wash buffer (300 µL / well) using a disk washer, and block the disk with 3% MSD blocking agent A (150 µL / well). Gently agitate the disk at ambient temperature and incubate for another 1–2 hours. Wash the disk three times with 1X MSD wash buffer using a disk washer. Add 30 µg of bone marrow lysate to the disk to a final volume of 50 µL / well. Add serially diluted standard samples to the same disk to a final volume of 50 µL / well. Incubate the disk overnight at 4°C with gentle agitation. Day 2

[0186] Wash the plates three times with 1X MSD wash buffer (prepared from 10X concentration buffer) using a plate washer. Add 50 µL / well of primary detection antibody (BTK RpAb; 1:1000 diluted in 1% MSD blocking agent A). Gently agitate the plates at room temperature and incubate for two hours. Wash the plates three times with 1X MSD wash buffer using a plate washer. Add 50 µL / well of anti-rabbit sulfonyl tag secondary antibody, diluted 1:500 in 1% MSD blocking agent A, and incubate the plates at room temperature for one to two hours. Wash the plates three times with 1X MSD tris wash buffer using a plate washer. Add 150 µL (1X) of MSD read buffer and read the plates using an MSD machine. Generate a standard curve and calculate the remaining BTK% in the medium relative to the treated sample using the absolute BTK number. Table 2 reports the remaining BTK in the samples.

[0187] The analysis results of Examples 1 to 19 are shown in the "Mouse Bone Marrow (%)" column of Table 2. Table 2 Example hWB BTK DC 50 (nM) hWB BTK dMax (%) BTK IC 50 (nM) CRBN IC 50 (nM) Mouse bone marrow (%) 1 4.08 99.99 0.0977 1260 11 2 5.93 98.84 0.128 749 12 3 2.88 99.24 0.0339 9570 twenty two 4 7.62 99.78 0.141 ≥10000 13 5 6.00 99.26 0.113 648 12 6 2.45 99.79 0.0403 378 19 7 18.7 99.34 0.0979 609 22 8 4.04 98.56 0.0804 3380 30 9 9.61 99.51 0.0988 658 31 10 3.04 98.5 0.106 6310 9 11 4.31 97.67 0.0973 2150 29 12 2.27 99.82 0.0363 2010 17 13 17.3 98.98 0.14 ≥10000 26 14 7.91 99.73 0.053 2330 16 15 4.21 99.91 0.0986 ≥10000 Nd 16 14.22 100.29 0.438 ≥10000 Nd 17 4.13 99.56 0.103 3870 Nd 18 15.6 99.5 0.496 1250 25 19 3 100 0.1 ≥10000 51 Nd = Undetermined compound 1 (Example 1) controls the generation of circulating leukemia burden tumor cells in C57BL / 6 mice by TCL1 carrying the BTKC481S mutation.

[0188] Eµ-TCL1CLL gene-transfected mice (Crogen Pharmaceuticals LLC) were crossed with BTK C481S knock-in mice (AbbVie) to generate Eµ-TCL1 gene-transfected mice carrying the BTK C481S mutation. Leukemia spleen cells from these BTK C481SEµ-TCL1 gene-transfected mice (generation 1) were intravenously transplanted into C57BL / 6 mice, resulting in CLL development in peripheral blood and spleen. When the circulating CLL burden (CD5+CD19+ cells as described below) in the recipient mice reached over 90%, the spleen cells containing high CLL burden (generation 2) were cryopreserved and used for CLL efficacy studies. Animal husbandry and management.

[0189] The study on the efficacy of tumor therapy in mice was conducted in accordance with AbbVie's Institutional Animal Care and Use Committee and the National Institutes of Health Guide for Care and Use of Laboratory Animals guidelines in the vivarium at South San Francisco, California. C57BL / 6 mice were obtained from Jackson Laboratories and housed in groups of 5 mice per cage under a 14-hour light:10-hour dark schedule. Tumor implantation and animal treatment were investigated.

[0190] Second-generation Eµ-TCL1BTK C481S spleen cells with high CLL load were thawed, washed, and filtered through a 70 µM filter in cold Hanks' Balanced Salt Solution (HBSS). 10 × 10⁶ spleen cells from 0.2 mL of HBSS were intravenously injected into the tail vein of 8-10 week old female C57BL / 6 mice to induce CLL development.

[0191] To determine the absolute / total white blood cell count, the complete blood cell count of each mouse was determined using a Genesis™ Veterinary Hematology Analyzer (Oxford Science Inc.). Flow cytometry

[0192] Cells isolated from mouse peripheral blood mononuclear cells (PMBCs) were collected to determine antigen expression. PMBCs were stained with antibodies specific to mouse CD5, CD19, and CD45 (cells expressing CD5+CD19+ are frequently found in mouse B-cell leukemia and are considered CLL cells). Cells were gated at CD45+, and the percentage of CD5+CD19+ cells from this subset was calculated. Flow cytometry was performed on a BD LSRFortessa™ X-20 cell analyzer (BD Biosciences).

[0193] Whole blood was collected via submandibular bleeding each week after tumor implantation. Circulating leukemia burden was calculated by multiplying the absolute / total white blood cell count by the percentage of white blood cells with the immunophenotype CD19+CD5+.

[0194] Starting 9 days post-inoculation, mice were weighed, bled, and randomly divided into five groups (N=10 for each group). The average circulating CD19+CD5+CLL cell count was 1,525 / µL. Compound 1 (Example 1) was prepared at different concentrations in a mediator containing 10% ethanol, 30% PEG-400, and 60% Phosal-50 MCT. The mediator contained 10% ethanol, 30% PEG-400, and 60% Phosal-50 MCT.

[0195] Two days after CLL count randomization, mice were treated for 19 days with the following once daily (QD): Group 1: mediator; Group 2: compound 1 dose 1; Group 3: compound 1 dose 2; Group 4: compound 1 dose 3; Group 5: compound 1 dose 4.

[0196] As shown in Figure 1, compound 1 (Example 1) dose-dependently controlled the circulating leukemia burden of TCL1-driven CLL carrying the BTK C481S mutation in C57BL / 6 mice. The y-axis shows the number of CLL cells (CD19+CD5+) per µL of mouse PMBC. Compound 1 achieved complete control of CLL burden (i.e., no increase from the initial tumor burden) at doses 3 and 4.

[0197] All publications and patents mentioned herein are incorporated herein by full reference, as if each individual publication or patent were specifically and individually indicated to be incorporated herein by reference.

[0198] It should be understood that the foregoing detailed description and accompanying examples are illustrative only and should not be regarded as a limitation on the scope of this disclosure, which is defined by the scope of the appended patent application and its equivalents. [Simplified Explanation of the Diagram]

[0028] [Figure 1] shows the in vivo activity of compound 1 (Example 1) in C57BL / 6 mice carrying the Btk C481S mutation in TCL1-driven CLL.

Claims

1. A compound, which is a system or a pharmaceutically acceptable salt thereof.

2. A compound which is a pharmaceutically acceptable salt.

3. A compound, its system.

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