Creatine kinase inhibitors

Abstract

The present invention relates to compounds of Formula (I): or a pharmaceutically acceptable salt thereof useful for treatment of a disease or disorder associated with CK and methods of inhibiting CK.

Description

CREATINE KINASE INHIBITORS GOVERNMENT LICENSE RIGHTS

[0001] This invention was made with government support under R21 CA259739 awarded by the National Institutes of Health. The government has certain rights in the invention. 5 RELATED APPLICATIONS

[0002] This application claims priority to US Provisional Application No.63 / 647,198, filed on May 14, 2024, the contents of which are hereby incorporated by reference in their entirety. FIELD

[0003] The present invention relates to inhibitors of creatine kinase useful for the treatment of 10 cancers. BACKGROUND

[0004] Creatine kinase (CK), also known as creatine phosphokinase (CPK) or phosphocreatine kinase is known to play a key role in cellular energy buffering and transport and provide local ATP production in periods of elevated energetic demand, such as during rapid anabolism and growth. CK 15 is essential for the growth and metastasis of aggressive acute myeloid leukemias, liver and breast cancer metastases, and pancreatic cancers. Because somatic tissues do not rely on CK for viability, the essentiality of CK for growth and metastasis of these cancers suggests CK inhibitor will be attractive targets for cancer therapy. Covalent CK inhibitors are described in WO2022087433 and Narek Darabedian, et al., “Depletion of creatine phosphagen energetics with a covalent creatine 20 kinase inhibitor, Nature Chemical Biology, 19, 815-824 (July 2023). New CK inhibitors are required to address an ongoing unmet need for new cancer therapies. SUMMARY OF THE INVENTION

[0005] An aspect of the invention is to provide compounds of Formula I, or a pharmaceutically acceptable salt thereof: 25where: A is phenyl or pyridinyl; R1is an optional substituent that, when present, is bonded to a ring carbon atom and is selected from the group consisting of C1 – C3 alkoxy, cyclopropoxy, amino, -NHCHO, and -NHC(O)CH3; X is -C(O)-, -C(O)NH-, or -CH2-; ;cycloalkyl). of Formula I, or a pharmaceutically acceptable salt thereof, wherein A is pyridinyl and R1is an optional substituent that, when present, is bonded to a ring carbon atom and is selected from the group consisting of C1 – C3 alkoxy, cyclopropoxy, amino, -NHCHO, and -NHC(O)CH3.

[0007] Another aspect of the invention is to provide compounds of Formula I, or a pharmaceutically acceptable salt thereof, wherein A is pyridinyl, R1is an optional substituent that, when present, is bonded to a ring carbon atom and is selected from the group consisting of C1– C3alkoxy, cyclopropoxy, amino, -NHCHO, and -NHC(O)CH3, and X is -C(O)-.

[0008] Another aspect of the invention is to provide compounds of Formula I, or a pharmaceutically acceptable salt thereof, wherein A is pyridinyl, R1is an optional substituent that, when present, is bonded to a ring carbon atom and is selected from the group consisting of C1 – C3 alkoxy,cyclopropoxy, amino, .

[0009] Anotheracceptable salt thereof, wherein A is pyridinyl, R1is an optional substituent that, when present, is bonded to a ring carbon atom and is selected from the group consisting of C1 – C3 alkoxy,cyclopropoxy, amino,R2is -CN.

[0010] Another aspect of the invention is to provide compounds of Formula I, or a pharmaceutically acceptable salt thereof, wherein A is phenyl, R1is an optional substituent that, when present, is bondedto a ring carbon atom and is selected from the group consisting of C1 – C3 alkoxy, cyclopropoxy,amino, -NHCHO, and -NHC(O)CH3, and .

[0011] Another aspect of the invention is I, or a pharmaceuticallyacceptable salt thereof, wherein A is phenyl, that, when present, is bonded to a ring carbon atom and is selected from the group consisting of C1– C3alkoxy, cyclopropoxy,amino,

[0012] Formula I, or a with amino, C1 – C3 alkoxy, cyclopropoxy, -NHCHO, or -NHC(O)CH3, X is -C(O)NH-, Y isprovides a pharmaceutical composition comprising a compound of Formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0014] A further aspect of the present invention provides a compound of Formula I or a pharmaceutically acceptable salt thereof for use in therapy.

[0015] Another aspect of the present invention provides a method for treating cancer susceptible to inhibition of CK comprising administering to a patient in need thereof a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof.

[0016] A further aspect of the present invention provides the use of a compound of Formula I or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of disorders susceptible to inhibition of CK.

[0017] Another aspect of the invention is the use of a compound of Formula I or a pharmaceutically acceptable salt thereof for the treatment of a cancer associated with creatine kinase overexpression. DETAILED DESCRIPTION

[0018] Terms used herein but not separately defined are taken to have their normal and customary meaning as understood by one of ordinary skill in the art.

[0019] The term “C1 – C4 alkyl” is taken to mean a straight or branched alkyl chain of from 1 to 4 carbon atoms and includes methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, and the like.

[0020] The term “C3 – C4 cycloalkyl” is taken to mean a carbocyclic ring containing 3 or 4 carbon 5 atoms and includes cyclopropyl and cyclobutyl, and the like.

[0021] The term “C1 – C3 alkoxy” is taken to mean a straight or branched alkyl chain of from 1 to 3 carbon atoms bonded to an oxygen atom and includes methoxy, ethoxy, propoxy, isopropoxy, and the like.

[0022] The term “patient” means mammal and “mammal” includes, but is not limited to, a human. 10

[0023] “Therapeutically effective amount” means the dosage of a compound of Formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing a compound of Formula I, or a pharmaceutically acceptable salt thereof, necessary to inhibit CK in a patient in need thereof. Anticipated dosages of a compound of Formula I, or a pharmaceutically acceptable salt thereof, are in the range of 1 mg / patient / day to 2000 mg / patient / day. The exact dosage required to 15 treat a patient and the duration of treatment will be determined by a physician in view of the stage and severity of the disease as well as the specific needs and response of the individual patient. Dosage administration may be adjusted to provide an optimal therapeutic benefit to an individual patient and to manage or avoid drug-related toxicities. For example, in addition to single daily dosing, multiple smaller daily doses or administration on a staggered daily, weekly, or monthly schedule may be 20 appropriate.

[0024] The terms “treatment”, “treat”, and “treating” are meant to include the full spectrum of pharmaceutical intervention for a patient in need of CK inhibition, such as administration of a CK inhibitor of the present invention to alleviate, slow, or reverse one or more of a patient’s symptoms or to delay progression of the disorder even if the disorder is not actually eliminated. 25

[0025] The compounds of Formula I are useful for the treatment of cancers associated with CK overexpression. These cancers include liver cancer, pancreatic cancer, lung cancer, colon cancer, and hematological cancers, including myelogenous leukemia, myeloid leukemia (including acute myeloid leukemia), myelodysplastic syndrome, lymphoblastic leukemia (including acute lymphoblastic leukemia), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), high risk 30 CLL, follicular lymphoma, diffuse large B-cell lymphoma (including DLBCL or ABC-DLBCL), mantle cell lymphoma (MCL), Waldenstrom’s macroglobulinemia (WM), multiple myeloma, marginal zone lymphoma (MZL), Burkitt’s lymphoma, non-Burkitt high grade B-cell lymphoma,extranodal marginal zone B-cell lymphoma, transformed high grade B-cell lymphoma (HGBL), lymphoplasmacytic lymphoma (LPL), central nervous system lymphoma (CNSL), or MALT lymphoma.

[0026] A further aspect of the invention is the use of a compound of Formula I for the treatment of acute myeloid leukemia (AML).

[0027] The compounds of Formula I are inhibitors of CK. The CK susceptible to inhibition of a compound of Formula I may be an isoform of creatine kinase such as CK-MM, CK-BB, or CK-MB.

[0028] Compounds of Formula I or a pharmaceutically acceptable salt thereof are preferably formulated as a pharmaceutical composition using a pharmaceutically acceptable carrier and administered by a variety of routes. Preferably, such compositions are for oral administration. Such pharmaceutical compositions and methods for preparing them are well known in the art. See, for example, REMINGTON: THE SCIENCE AND PRACTICE OF PHARMACY (L.V. Allen ed., Pharmaceutical Press, 22ndEdition, 2012).

[0029] A compound of Formula I or a pharmaceutically acceptable salt thereof may be administered either simultaneously with, or before, or after, one or more other therapeutic agents. When said compounds or pharmaceutically acceptable salts thereof are administered with one or more therapeutic agents, they may be administered separately, by the same or different route of administration, or together in the same pharmaceutical composition as the other therapeutic agent or agents. Where one or more additional therapeutic agents are administered, the administration of each therapeutic agent may be simultaneous, separate, or sequential.

[0030] Compounds of Formula I are capable of reacting with a number of inorganic and organic acids to form pharmaceutically acceptable acid addition salts. Such pharmaceutically acceptable salts and common methodology for preparing them are well known in the art. See, for example, P. Stahl, et al., HANDBOOK OFPHARMACEUTICALSCIENCES, (VCHA / Wiley – VCH, 2002); S.M. Berge, et al., Pharmaceutical Salts, 66 JOURNAL OF PHARMACEUTICAL SCIENCES 1 (1977).

[0031] It will be understood that compounds of Formula I have a chiral center and may be depicted as single stereoisomers. For example, a compound of Formula I is illustrated below:5

[0032] All stereoisomers of the compounds of Formula I are contemplated within the scope of the present invention. As used herein, references to a single stereoisomer are meant to also include stereoisomeric mixtures including the named or depicted compound of Formula I. Herein, theCahn-Ingold-Prelog designations of (R)- and (S)- may be used to refer to specific stereoisomers. Specific stereoisomers can be prepared by stereospecific synthesis using enantiomerically pure or enriched starting materials. The specific stereoisomers of either starting materials, intermediates, or racemic mixtures including compounds of Formula I and II can be resolved by techniques well 5 known in the art, such as those found in Stereochemistry of Organic Compounds, E. I. Eliel and S. H. Wilen (Wiley 1994) and Enantiomers, Racemates, and Resolutions, J., Jacques, A Collet, and S. H. Wilen (Wiley 1991), including chromatography on chiral stationary phases, enzymatic resolutions, or fractional crystallization or chromatography of diastereomers formed for that purpose, such as diastereomeric salts. 10

[0033] It will be further understood that when A is pyridinyl, the following four isomers are contemplated:and when the term pyridinyl is used herein with respect to ring A it is taken to mean all four isomers unless a specific isomer is identified. 15

[0034] It should be understood that the compounds described herein, unless otherwise specified, encompass and include all isomers and stable isotopic variants of the compounds of the present disclosure, such as deuterated compounds. All isotopic variants of the compounds provided herein, whether radioactive or not, should be included within the scope of the present disclosure.

[0035] Compounds of the present invention may be named according to IUPAC, and may also be 20 named according to CAS, and other naming conventions may be used to unambiguously identify a compound of Formula I or a pharmaceutically acceptable salt thereof.

[0036] The compounds employed as initial starting materials in the synthesis of compounds of Formula I are well known and, to the extent not commercially available, are readily synthesized using specific references provided, by standard procedures commonly employed by those of ordinary skill 25 in the art, or are found in general reference texts. Examples of known procedures and methods include those in general reference texts such as: COMPREHENSIVE ORGANIC TRANSFORMATIONS (VCHPublishers Inc., 1989); COMPENDIUM OFORGANICSYNTHETICMETHODS(Wiley Interscience, Volumes 1 – 10, 1974 – 2002); Michael B. Smith and Jerry March, ADVANCED ORGANIC CHEMISTRY, REACTIONS, MECHANISMS, AND STRUCTURE (Wiley Interscience, 5thed.2001); Francis A. Carey andRichard J. Sundberg, ADVANCEDORGANICCHEMISTRY, PARTB, REACTIONS ANDSYNTHESIS(KluwerAcademic / Plenum Publishers, 4thed.2000), and references cited therein.

[0037] Certain intermediates described in the following preparations may contain one or more nitrogen protecting groups. It is understood that protecting groups may be varied as appreciated by one of skill in the art depending on the actual reaction conditions and the transformations to be performed. The protection and deprotection conditions are well known to the skilled artisan and are described in the literature (See for example "Greene's Protective Groups in Organic Synthesis", Fifth Edition, by Peter G.M. Wuts and Theodora W. Greene, John Wiley and Sons, Inc.2014).

[0038] Compounds of Formula I or pharmaceutically acceptable salts thereof may be prepared by a variety of procedures known in the art, some of which are illustrated in the Schemes, Preparations, and Examples below which are provided to further illustrate the invention without limiting the scope of the invention in any way. The specific steps and methodology for each of the synthetic routes described may be combined in different ways, or in conjunction with steps from different schemes, to prepare compounds of the present invention or pharmaceutically acceptable salts thereof. The products of each step in the schemes below may be isolated by conventional methods well known in the art, including extraction, evaporation, precipitation, chromatography, filtration, trituration, and crystallization. In the schemes below, all substituents are as previously defined unless otherwise indicated.

[0039] The abbreviations used herein are defined according to Aldrichimica Acta, vol.17, No.1, 1984. Other abbreviations are defined as follows: “ACN” refers to acetonitrile; “AcOH” refers to acetic acid; “BF3.Et2O” refers to boron trifluoride etherate; “(Boc)2O” refers to di-tert-butyl decarbonate; “cataCXium A Pd G3” refers to [(di(1-adamantyl)-butylphosphine)-2-(2′-amino-1,1′- biphenyl)]palladium(II) methanesulfonate; “DBU” refers to 1,8-diazabicyclo[5.4.0]undec-7-ene; “DCC” refers to N,N′-dicyclohexylcarbodiimide; “DCM” refers to dichloromethane; “DIEA” refers to diisopropylethylamine; “DMAP” refers to 4-dimethylaminopyridine; “DMF” refers to dimethylformamide; “DMP” refers to Dess–Martin periodinane; “EDCI” refers to 1-ethyl-3-(3- dimethylaminopropyl)carbodiimide; “EtOAc” refers to ethyl acetate; “EtOH” refers to ethanol; “HATU” refers to 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate; “h” refers to hour(s); “LAH” refers to lithium aluminium hydride; “m-CPBA”refers to meta-chloroperoxybenzoic acid; “MeOH” refers to methanol; “NaBH3CN” refers to sodium cyanoborohydride; “NaOAc” refers to sodium acetate; “NaSME” refers to sodium methanethiolate; “n-BuLi” refers to n-butyllithium; “Pd(t-Bu3P)2” refers to bis(tri-tert-butylphosphine)palladium(0); “PG” refers to protecting group; “PhOH” refers to phenol; “prep-HPLC” refers to preparative high 5 performance liquid chromatography; “prep-TLC” refers to preparative thin layer chromatography; “Py” refers to pyridine; “TBME” refers to tert-butyl methyl ether; “t-BuOH” refers to tert-butanol; “TEA” refers to triethylamine; “TFA” referstotrifluoroacetic acid; “THF” refers to tetrahydrofuran;“TsCl” refers to 4-toluenesulfonyl chloride.

[0040] In the schemes below, all substituents unless otherwise indicated,are aspreviously defined. 10 The reagents and starting materials are either commercially available or may be prepared by methods well known to one of ordinary skill in the art, some of which are presented in the preparations below. Without limiting the scope of the invention, the following schemes, preparations, and examples are provided to further illustrate the invention. 15 Scheme 1

[0041] X is -C(O)-beginning with a suitable acid (i) and amine (ii). Amide coupling conditions are well known to the skilled person and include reacting a solution of the carboxylic acid (i) and a desired amine (ii) in a 20 suitable solvent, such as DMF, with an appropriate coupling reagent, such as HATU, in the presence of a suitable organic base, such as DIEA or TEA, with stirring at ambient temperature for at least 2 h.Scheme 2

[0042] where X is - CH2- under standard reductive amination conditions. A suitable aldehyde (iii) is reacted with the 5 amine (ii) in the presence of a reducing agent, such as NaBH3CN, at a suitable pH adjusted between 5 and 6 with AcOH, in an appropriate solvent, such as MeOH, at 25 °C for at least 1 h. Scheme 3 10

[0043] where X is - C(O)NH- under urea formation conditions well known to a skilled artisan. The suitable amines (iv) and (ii) were reacted in the presence of triphosgene and an appropriate base, such as TEA, in a suitable solvent, such as DCM.

[0044] The skilled artisan will appreciate that protecting groups employed in the preparation of 15 compounds of Formula I may be removed as necessary or desired at any convenient point in the synthesis.PREPARATIONS Preparation 1 tert-Butyl (2,3-dioxoindolin-4-yl)carbamate

[0045] To a solution of 4- g, 16.4 mmol) and tert-butyl carbamate(11.51 g, 98.22 mmol) in dioxane (50 (10.67 g, 32.74 mmol) and XPhos Pd G3 (2.77 g, 3.27 mmol). The mixture was stirred at 80 °C for 12 h under N2atmosphere. The reaction mixture was concentrated under reduced pressure to remove solvent. The reaction mixture was diluted with H2O (20 mL), and then extracted with EtOAc (3 x 20 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, 0- 16% EtOAc / Petroleum ether gradient @ 90 mL / min) to give the title compound (2.2 g, 8.4 mmol, 51%) as a yellow solid.

[0046] 1H NMR (400 MHz, DMSO-d6) δ (ppm) 11.20 - 10.92 (m, 1H), 8.84 - 8.56 (m, 1H), 7.78 - 7.44 (m, 2H), 6.49 (d, J = 7.4 Hz, 1H), 1.55 - 1.45 (m, 9H) Preparation 2 (E)-3-(Hydroxyimino)-1,3-dihydro-2H-pyrrolo[2,3-b]pyridin-2-one

[0047] To a solution of 1H-pyrrolodione (4.6 g, 31.1 mmol) in H2O (60 mL) was added NH2OH.HCl (2.27 g, 32.61 mmol) and the mixture was stirred at 100 °C for 0.5 h. Then NaOAc (2.29 g, 27.95 mmol) was added and the mixture was stirred at 100 °C for 0.5 h. The reaction mixture was filtered and the cake was washed with H2O (20 mL) and dried to give the title compound (4.2 g, 24.2 mmol, 78%) as a red solid.

[0048] 1H NMR (400 MHz, DMSO-d6) δ (ppm) 7.03 (dd, J = 7.38, 5.38 Hz, 1 H) 8.12 - 8.20 (m, 2 H) 11.31 (br s, 1 H) 13.58 (s, 1 H)

[0049] The compound in the following table was prepared essentially as described in Preparation 2. Preparation Chemical name Structure tert-Butyl (Z)-(3-(hydroxyimino)-2- 3 oxoindolin-4-yl)carbamate Preparation 4 tert-Butyl (2-oxo-2,3-dihydro-1H-pyrrolo[2,3-b]pyridin-3-yl)carbamate 5

[0050] To a solution of (E)-3- 2H-pyrrolo[2,3-b]pyridin-2-one (4.2 g,25.8 mmol) and (Boc)2O (11.24 g, 51.49 in MeOH (80 mL) was added Pd / C (4.11 g, 3.86 mmol). The mixture was stirred at 25 °C for 4 h under H2 (15 psi). Then Pd / C (4.11 g, 3.86 mmol) was added and the mixture was stirred at 25 °C for 4 h under H2 (15 psi). The reaction mixture 10 was filtered and the filtrate was concentrated to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0-100% EtOAc / Petroleum ether gradient @ 150 mL / min) and then triturated with EtOAc (20 mL). The resulting residue was collected by filtration, washed with EtOAc (5 mL) and dried to give the title compound (1.3 g, 4.9 mmol, 19%) as a pink solid. 15

[0051] ESI-MS (m / z) = 250 (M+H).Preparation 5 Di-tert-butyl (2-oxoindoline-3,4-diyl)dicarbamate

[0052] To a solution of tert-butyl -2-oxoindolin-4-yl) carbamate (2 g, 7mmol) in EtOH (20 mL) was added 2O (3.15 g, 14.43 mmol, 3.31 mL). The mixture was stirred at 20 °C for 1.5 h under H2 (15 Psi). The reaction mixture was concentrated under reduced pressure to remove solvent. The reaction mixture was diluted with H2O (20 mL), and then extracted with EtOAc (3 x 20 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0-40% EtOAc / Petroleum ether gradient @ 80 mL / min) to give the title compound (1 g, 3 mmol, 38%) as a yellow solid.

[0053] 1H NMR (400 MHz, DMSO-d6) δ (ppm) 10.62 - 10.25 (m, 1H), 8.28 - 8.05 (m, 1H), 7.75 - 7.46 (m, 2H), 7.13 (br t, J = 8.0 Hz, 1H), 6.48 (d, J = 7.6 Hz, 1H), 4.97 (br d, J = 8.3 Hz, 1H), 1.52 - 1.38 (m, 18H) Preparation 6 tert-Butyl (2,3-dihydro-1H-pyrrolo[2,3-b]pyridin-3-yl)carbamate

[0054] To a solution of tert-butyl1H-pyrrolo[2,3-b]pyridin-3-yl)carbamate (500 mg, 2 mmol) in THF (10 mL) was added LAH (2.5 M, 1.60 mL) at 0 °C under N2 atmosphere and the mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated. The residue was purified by prep-TLC (SiO2, 50% EtOAc in Petroleum ether) to give the title compound (110 mg, 468 μmol, 23%).

[0055] ESI-MS (m / z) = 236 (M+H).Preparation 7 Di-tert-butyl indoline-3,4-diyldicarbamate

[0056] To a mixture of di-tert- diyl)dicarbamate (600 mg, 2 mmol) andBF3.Et2O (468.66 mg, 3.30 mmol,was added LiAlH4 (2.5 M, 1.32 mL) dropwise at 0 °C under N2 atmosphere, stirred at 0 °C for 1 h. Then the mixture was stirred at 70 °C for 3 h. The reaction mixture was quenched by added 10.H2O Na2SO4(300 mg) at 0 °C and stirred at 0 °C for 1 h. Then the reaction mixture was diluted with H2O (10 mL), and then extracted with EtOAc (3 x 10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give the title compound (500 mg) as a brown oil.

[0057] ESI-MS (m / z) = 372 (M+Na). Preparation 8 4-Methoxy-3-nitro-1H-indole

[0058] To a solution of 4-methoxy-1H-g, 34.7 mmol) in ACN (150 mL) was added AgNO3 (6.18 g, 36.39 mmol) at 25 °C under N2 atmosphere. Then the reaction mixture was cooled to 0 °C, benzoyl chloride (5.11 g, 36.39 mmol, 4.22 mL) was added dropwise to the reaction mixture over 0.5 h at 0 °C and the reaction mixture was stirred at 0 °C for 0.5 h under N2atmosphere. The reaction mixture was concentrated to give a residue. The residue was diluted with H2O (30 mL) and extracted with EtOAc (3 x 100 mL). The combined organic layers were dried over Na2SO4, filtered and the filtrate was concentrated to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash ® Silica Flash Column, 0-16% EtOAc / Petroleum ether gradient @ 100 mL / min) to give the title compound (1 g, 5 mmol, 15%) as a yellow oil.Preparation 9 tert-Butyl 4-methoxy-3-nitro-1H-indole-1-carboxylate

[0059] To a solution of 4-methoxy-3- g, 10 mmol) in THF (20 mL) were addedTEA (3.16 g, 31.22 mmol, 4.35 mL), tert-butyl carbonate (4.54 g, 20.81 mmol, 4.78 mL) and DMAP (127.14 mg, 1.04 mmol), the mixture was stirred at 20 °C for 12 h. The reaction mixture was concentrated to give a residue. The residue was diluted with H2O (20 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layers were dried over Na2SO4, filtered and the filtrate was concentrated to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash ® Silica Flash Column, 0-10% EtOAc / Petroleum ether gradient @ 80 mL / min) to give the title compound (1.3 g, 4.5 mmol, 43%) as a white solid. Preparation 10 N-(2-Bromo-6-nitrophenyl)-4-methylbenzenesulfonamide

[0060] To a solution of 2-bromo-6-mmol) in THF (250 mL) was added NaH (7.32 g, 182.89 mmol) at 0 °C under N2atmosphere. After addition, the mixture was stirred at this temperature for 0.5 h, and then TsCl (26.15 g, 137.16 mmol) was added at 0 °C. The resulting mixture was stirred at 0 °C for 0.5 h, then warmed to 20 °C and stirred at 20 °C for 1 h. The mixture was then warmed to 80 °C and was stirred at 80 °C for 12 h. The reaction mixture was quenched by saturated NH4Cl solution (500 mL) and extracted with EtOAc (3 x 500 mL). The combined organic phase was concentrated to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 220 g SepaFlash®Silica Flash Column, 0-15% EtOAc / Petroleum ether gradient @ 200 mL / min) to give the title compound (20 g, 44 mmol, 49%) as a yellow solid.Preparation 11 4-Methyl-N-(2-nitro-6-vinylphenyl)benzenesulfonamide

[0061] To a solution of N-(2- methyl-benzenesulfonamide (18 g, 49 mmol) in toluene (180 mL) was (23.06 g, 72.74 mmol, 21.24 mL) under N2atmosphere, then Pd(t-Bu3P)2(1.224 g, 2.42 mmol) was added under N atmosphere at 20 °C. The mixture was stirred at 120 °C for 4 h. The reaction mixture was quenched by KF solution (200 mL). The residue was diluted with H2O (100 mL) and extracted with EtOAc (3 x 300 mL). The combined organic layers were dried over Na2SO4, filtered and the filtrate was concentrated to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash®Silica Flash Column, Eluent of 0-15% EtOAc / Petroleum ether gradient @ 80 mL / min) to give the title compound (17 g, 16 mmol, 32%) as a yellow solid. Preparation 12 N-Benzyl-1-tosyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-3-amine

[0062] To a solution of 4-methyl-yl)benzenesulfonamide (2 g, 7 mmol) in ACN (20 mL) was added phenylmethanamine (1.56 g, 14.58 mmol, 1.59 mL), KI (1.21 g, 7.29 mmol) and [acetoxy(phenyl)-iodanyl] acetate (3.52 g, 10.94 mmol). The mixture was stirred at 20 °C for 18 h. The reaction mixture was diluted with H2O (100 mL), and then extracted with DCM (4 x 50 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40g SepaFlash® Silica Flash Column, 0-60% EtOAc / Petroleum ether gradient @ 80 mL / min) to give the title compound (1.1 g, 2.5 mmol, 34%) as a white solid.

[0063] The compound in the following table was prepared essentially as described in Preparation 12.Preparation Chemical name Structure 13 N-Benzyl-7-nitro-1-tosylindolin-3-amine Preparation 14 tert-Butyl 3-amino-4-methoxyindoline-1-carboxylate

[0064] To a solution of tert-butyl 4-indole-1-carboxylate (500 mg, 2 mmol) in 5 2,2,2-trifluoroethanol (3 mL) was added Pd / C (2.80 g) at 25 °C. The suspension was degassed and purged with H2for 3 times. The mixture was stirred at 30 °C for 12 h under H2(342.13 μmol) (15 psi). The reaction mixture was concentrated to give the title compound (400 mg) as a yellow oil.

[0065] The compounds in the following table were prepared essentially as described in Preparation 14. Preparation Chemical name Structure 15 1-Tosyl-2,3-dihydro-1H-pyrrolo[3,2- b]pyridin-3-amine 161-Tosylindoline-3,7-diaminePreparation 17 tert-Butyl (1-tosyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-3-yl)carbamate

[0066] To a solution of 1-tosyl-2,3-[3,2-b]pyridin-3-amine (400 mg, 1 mmol) 5 in DCM (10 mL) was added (Boc)2O mg, 476 μL) and TEA (167.86 mg, 1.66 mmol, 231 μL). The mixture was stirred at 20 °C for 12 h. The reaction mixture was diluted with Na2SO3 saturated (100 mL) and then extracted with DCM (4 x 50 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40g SepaFlash® Silica Flash 10 Column, 0-30% EtOAc / Petroleum ether gradient @ 80 mL / min) to give the title compound (250 mg, 520 μmol, 38%) as a white solid.

[0067] The compound in the following table was prepared essentially as described in Preparation 17. Preparation Chemical name Structure 18 tert-Butyl (7-amino-1-tosylindolin-3- yl)carbamate 15Preparation 19 tert-Butyl (7-formamido-1-tosylindolin-3-yl)carbamate

[0068] To a solution of tert-butyl 3-yl)carbamate (350 mg, 867 μmol) in 5 DCM (2 mL) were added EDCI HCOOH (45.84 mg, 954.16 μmol) at 20 °C and the mixture was stirred at 20 °C for 2 h. The reaction mixture was concentrated to give a residue. The residue was dissolved inACN (2.5 mL), and the resulting solution was purified by prep- HPLC (TFA condition, column: Phenomenex Luna C18100*30mm*5um; mobile phase: [H2O(0.1% TFA)-ACN]; gradient: 35%- 65% B over 8.0 min) to give the title compound (250 mg, 579 μmol, 10 67%) as a white solid. Preparation 20 tert-Butyl (7-formamidoindolin-3-yl)carbamate

[0069] To a solution of1.14 mL) in dry THF (3 mL) was 15 added Na (319.67 mg, 13.90 mmol, 330 μL) a 25 °C and the mixture was treated by sonication for 30 min. A solution of tert-butyl (7-formamido-1-tosylindolin-3-yl)carbamate (200 mg, 464 μmol) in dry THF (3 mL) was added to the mixture at -78 °C. The resulting mixture was stirred at -78 °C for 5 min. The reaction mixture was quenched b H2O (10 mL) and extracted with TBME (3 x 10 mL). The combined organic phase was washed with brine (5 mL), dried over anhydrous Na2SO4, filtered 20 and concentrated give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® Silica Flash Column, 0-65% EtOAc / Petroleum ether gradient @ 40 mL / min) to give the title compound (80 mg, 242 52%) as a brown solid.Preparation 21 tert-Butyl (2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-3-yl)carbamate

[0070] To a solution of tert-butyl 1H-pyrrolo[3,2-b]pyridin-3-yl)carbamate(300 mg, 770 μmol) in MeOH (5 mL) added Mg (280.83 mg, 11.55 mmol) and NH4Cl (123.61 mg, 2.31 mmol). The mixture was stirred at 20 °C for 1h. The reaction mixture was diluted with H2O (10 mL), and then extracted with DCM (4 x 5 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give the title compound (180 mg) as a white solid. Preparation 22 tert-Butyl (1-acryloylindolin-3-yl)carbamate (racemic)

[0071] To a solution of tert-butyl(50 mg, 213 μmol) in DCM (3 mL) was added TEA (43.19 mg, 426.82 μmol, 59 μL). Then acryloyl chloride (21.25 mg, 234.75 μmol, 19 μL) was added at 0 °C. The mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by prep-TLC (SiO2, 50% EtOAc in Petroleum ether) to give the title compound (70 mg, 202 μmol, 95% yield) as yellow oil.

[0072] ESI-MS (m / z) = 233 (M+H-56).

[0073] The compounds in the following table were prepared essentially as described in Preparation 22. Prep Chemical name Struct ESI-MS No. ure (m / z) 23 tert-Butyl (1-acryloylindolin-3- yl)carbamate (Isomer 1) N / A tert-Butyl (1-ac 426 24 ryloylindolin-3- yl)carbamate (Isomer 2) (M+Na) Preparation 25 tert-Butyl (1-acryloyl-2,3-dihydro-1H-pyrrolo[3,2-b]pyridin-3-yl)carbamate 5

[0074] To a solution of tert-butyl[3,2-b]pyridin-3-yl)carbamate (180 mg, 765 μmol) in DCM (2 mL) was added TEA (77.41 mg, 765.04 μmol, 107 μL) and prop-2-enoyl chloride (69.24 mg, 765.04 μmol, 62 μL). The mixture was stirred at 20 °C for 1 h. The reaction mixture was diluted with Na2SO3saturated solution (10 mL), and then extracted with DCM (4 x 5 10 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 4g SepaFlash® Silica Flash Column, Eluent of 0-30% EtOAc / Petroleum ethergradient @ 80 mL / min) to give the title compound (20 mg, 56 μmol, 7%) as a white solid.

[0075] 1H NMR (400 MHz, DMSO-d6) δ (ppm) 10.62 - 10.25 (m, 1H), 8.28 - 8.05 (m, 1H), 7.75 - 7.46 (m, 2H), 7.13 (br t, J = 8.0 Hz, 1H), 6.48 (d, J = 7.6 Hz, 1H), 4.97 (br d, J = 8.3 Hz, 1H), 1.52 - 1.38 (m, 18H)

[0076] The compounds in the following table were prepared essentially as described in Preparation 5 25.PrepChemical na ESI-MS No. me Structure (m / z)tert-Butyl (1-acryloyl-2,3-dihydro- 26 1H-pyrrolo[2,3-b]pyridin-3- N / A yl)carbamateDi-tert-butyl (1-acryloylindoli 426 27 ne- 3,4-diyl)dicarbamate(M+Na)28 tert-Butyl (1-acryloyl-7- formamidoindolin-3-yl)carbamateN / Preparation 29 5-Bromo-2-(methylthio)benzo[d]thiazole

[0077] To a solution of 5-bromo-2-chlorobenzo[d]thiazole (1.5 g, 6.0 mmol) in EtOH (30 mL) was added NaSMe (0.85 mg, 12.13 μmol, 1 μL). The mixture was stirred at 20 °C for 2 h. The reaction mixture was diluted with H2O (20 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with NaCl saturated solution (4 x 20 mL), dried over, filtered and concentrated under reduced pressure to give the title compound (1.5 g, 5.8 mmol, 96%) as a white solid.

[0078] ESI-MS (m / z) = 260 (M+H). Preparation 30 2-(Methylthio)benzo[d]thiazole-5-carboxylic acid

[0079] To a solution of 5-bromo-thiazole (500 mg, 2 mmol) in THF (10 mL) was added n-BuLi (2.5 M, 1.15 mL) at -70 °C. The mixture was stirred for 20 min. The dry ice was added to the mixture and stirred at -70 °C for 1 h. The reaction mixture was quenched by addition HCl (1 M) 20 mL at 0 °C, and then extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with NaCl saturated solution (3 x 20 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the title compound (400 mg, 2 mmol, 31%) as a yellow solid.

[0080] ESI-MS (m / z) = 226 (M+H). Preparation 31 2-(Methylsulfonyl)benzo[d]thiazole-5-carboxylic acid

[0081] To a solution of 2-carboxylic acid (100 mg, 444 μmol) in DCM (5 mL) was added m-CPBA (210.64 mg, 976.53 μmol). The mixture was stirred at 25 °C for 2 h. The reaction mixture was quenched by addition Na2SO3(5 mL) at 0 °C, and then extracted with DCM / MeOH (v / v = 5 / 1) (3 x 5 mL). The combined organic layers were washed with NaCl saturated solution (3 x 3 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Luna C18100 * 30 mm * 5um; mobile phase: [H2O (0.1% TFA) - ACN]; gradient: 15% - 45% B over 8.0 min) to give the title compound (270 mg, 1 mmol, 59%) as a light yellow solid.

[0082] ESI-MS (m / z) = 258 (M+H). Preparation 32 tert-Butyl 4-bromo-3-nitrobenzoate

[0083] A mixture of 4-bromo-3-mmol), DMAP (993.17 mg, 8.13 mmol), and DCC (5.87 g, 28.45 mmol, was degassed and purged with N2 (3 times), and then t-BuOH (7.68 g, 103.65 mmol, 9.91 mL) was added at 0 °C. This mixture was stirred at 25 °C for 12 h under N2 atmosphere. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel column chromatography with EtOAc and petroleum ether to obtain the title compound (2 g, 7 mmol, 33%) as a white solid.

[0084] 1H NMR (400 MHz, CDCl3) δ 8.37 (d, J = 2.0 Hz, 1H), 8.01 (dd, J = 1.9, 8.3 Hz, 1H), 7.81 (d, J = 8.3 Hz, 1H), 1.61 (s, 9H) Preparation 33 tert-Butyl 3-amino-4-bromobenzoate

[0085] To a solution of tert-butyl(2 g, 7 mmol) in EtOH (15 mL) and H2O (5 mL) was added Fe (1.85 g, 33.10 mmol) and NH4Cl (1.77 g, 33.10 mmol). The mixture was stirred at 85 °C for 2 h. The residue was diluted with H2O (10 mL) and extracted with EtOAc (3 x 10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography with EtOAc and petroleum ether to obtain the title compound (1.56 g, 5.73 mmol, 87%) as a white solid.

[0086] 1H NMR (400 MHz, CDCl3) δ 7.44 (d, J = 8.3 Hz, 1H), 7.38 (d, J = 2.0 Hz, 1H), 7.22 (dd, J = 2.0, 8.3 Hz, 1H), 4.24 - 4.14 (m, 2H), 1.58 (s, 9H).Preparation 34 tert-Butyl (Z)-4-bromo-3-((4-chloro-5H-1,2,3-dithiazol-5-ylidene)amino)benzoate

[0087] A mixture of tert-butyl 3- mg, 184 μmol) and 4,5-dichloro-5 1,2,3-dithiazolium chloride (45.97(3 mL) was degassed and purged with N2 (3 times), and then the mixture was at 20 °C for 1 h under N2 atmosphere. DBU (55.94 mg, 367.46 μmol, 55 μL) was added at 0 °C and the mixture was stirred at 20 °C for 0.5 h. Then the mixture was stirred at 40 °C for 5 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography with 10 EtOAc and petroleum ether to obtain the title compound (637 mg, 2 mmol, 79%) as a yellow oil.

[0088] ESI-MS (m / z) = 407 (M+H). Preparation 35 tert-Butyl 2-cyanobenzo[d]thiazole-5-carboxylate 15

[0089] A solution of3-((4-chloro-5H-1,2,3-dithiazol-5- ylidene)amino)benzoate (300 mg, 736 μmol) and CuI (210.19 mg, 1.10 mmol) in Py (10 mL) was taken up into a microwave tube and the sealed tube was heated at 115 °C for 1 h under microwave irradiation. The reaction mixture was concentrated under reduced pressure to remove solvent and the residue was diluted with H2O (5 mL) and extracted with EtOAc (3 x 5 mL). The combined organic20layers were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography with EtOAc and petroleum ether to obtain the title compound (200 mg, 636 μmol, 86%) as a yellow oil.

[0090] ESI-MS (m / z) = 261 (M+H).Preparation 36 1-(3-Aminoindolin-1-yl)prop-2-en-1-one (racemic)

[0091] To a solution of tert-butyl (1- yl)carbamate (50 mg, 173 μmol) in DCM (25 mL) was added TFA (197.72 mg, 1.73 . The mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure to obtain the title compound (50 mg) as a yellow oil.

[0092] The compounds in the following table were prepared essentially as described in Preparation 36. Prep Chemical name Structur ESI-MS No. e (m / z) 37 2-Cyanobenzo[d]thiazole-5- carboxylic acid N / A1-(3-Amino-2,3-dihydro-1H-38 pyrrolo[3,2-b]pyridin-1-yl)prop-2- N / A en-1-one 1-(3-Amino-2,3-dihydro-1H- 39 pyrrolo[2,3-b]pyridin-1-yl)prop-2- N / A en-1-one 1-(3,4-Diaminoindolin-1- 187 40 yl)prop- 2-en-1-one[M-NH3]-26-41 N-(1-Acryloyl-3-aminoindolin-7- yl)formamide N / A 42 1-(3-Aminoindolin-1-yl)prop-2- en-1-one (Isomer 1) N / A 43 1-(3-Aminoindolin-1-yl)prop-2- en-1-one (Isomer 2) N / A Preparation 44 Methyl 2-phenoxybenzo[d]thiazole-5-carboxylate

[0093] To a solution of PhOHμL) in ACN (4 mL) was added K2CO35 (364.23 mg, 2.64 mmol), stirred at 20 °C for 1 h. Then methyl 2-chlorobenzo[d]thiazole-5- carboxylate (300 mg, 1 mmol) was added to the mixture. The mixture was stirred at 80 °C for 12 h. The reaction mixture was diluted with H2O (5mL), and then extracted with EtOAc (3 x 5mL). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g 10 SepaFlash® Silica Flash Column, 0-3% EtOAc / Petroleum ether gradient @ 70 mL / min) to give the title compound (320 mg, 1 mmol, 85%) as a white solid.

[0094] ESI-MS (m / z) = 286 (M+H). Preparation 45 2-Phenoxybenzo[d]thiazole-5-carboxylic acid 15

[0095] To a solution of methyl 2-phenoxybenzo[d]thiazole-5-carboxylate (300 mg, 1 mmol) in THF (3.2 mL), H2O (0.8 mL) was added LiOH.H2O (220.62 mg, 5.26 mmol). The mixture was stirred at 20 °C for 12 h. The reaction mixture was diluted with H2O (2 mL), and then extracted with EtOAc (2 mL). The organic phase was discarded. The aqueous phase was adjusted to pH = 5 with 1N HCl and the aqueous phase extracted with EtOAc (3 x 3 mL). The combined organic phase was dried with anhydrous Na2SO4, the mixture was filtered and the filtrate was concentrated in vacuo to give the title compound (280 mg) as a white solid.

[0096] ESI-MS (m / z) = 272 (M+H) Preparation 46 6-Hydroxymethyl)benzo[d]thiazole-2-carbonitrile

[0097] A mixture of 6-(500 mg, 2 mmol), tributylstannylmethanol (1.34 g, 4.18 mmol) and cataCXium A Pd G3 (152.30 mg, 209.12 μmol) in dioxane (10 mL) was degassed and purged with N2(3 times), and then the mixture was stirred at 60 °C for 12 h under N2 atmosphere. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by flash silica gel chromatography (0-40% EtOAc in Petroleum ether @ 100 mL / min) to give the title compound (310 mg, 1 mmol, 62%) as a yellow solid.

[0098] 1H NMR (400 MHz, DMSO-d6) δ 8.27 (d, J = 0.9 Hz, 1H), 8.21 (d, J = 8.6 Hz, 1H), 7.66 (dd, J = 1.6, 8.6 Hz, 1H), 5.52 (t, J = 5.7 Hz, 1H), 4.70 (d, J = 5.8 Hz, 2H). Preparation 47 6-Formylbenzo[d]thiazole-2-carbonitrile

[0099] To a solution of 6-2-carbonitrile (100 mg, 526 μmol) in DCM (3 mL) was added DMP (245.27 mg, 578.27 μmol, 179 μL). The mixture was stirred at 25 °C for 1 h. The residue was diluted with H2O (5 mL) and extracted with DCM (3 x 5 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give the title compound (100 mg) as a yellow solid.Preparation 48 tert-Butyl 3-(2-cyanobenzo[d]thiazole-5-carboxamido)-4-methoxyindoline-1-carboxylate

[0100] To a solution of 2-cyano-acid (154.51 mg, 756.66 μmol) in 5 DMF (1 mL) were added HATU mg, and DIEA (195.59 mg, 1.51 mmol, 264 μL)and the mixture was stirred at 20 °C for 0.5 h. tert-butyl 3-amino-4-methoxy-indoline-1- carboxylate (200 mg, 757 μmol) was added and the reaction mixture was stirred at 20 °C for 1 h. The reaction mixture was diluted with DMF (2 mL), and the resulting solution was purified by prep-HPLC (column: Waters Xbridge Prep OBD C18150 * 40 mm * 10 um; mobile phase: [H2O (10 mM 10 NH4HCO3) - ACN]; gradient: 50% - 80% B over 8.0 min) to give the title compound (30 mg, 67 μmol, 9%) as a white solid. Preparation 49 2-Cyano-N-(4-methoxyindolin-3-yl)benzo[d]thiazole-5-carboxamide 15

[0101] To a solution of tert-5-carbonyl)amino]-4-methoxy- indoline-1-carboxylate (30 mg, 67 μmol) in DCM (1 mL) was added TFA (22.78 mg, 199.77 μmol, 15 μL), the mixture was stirred at 20 °C for 1h. The reaction mixture was concentrated to give the title compound (30 mg, TFA salt) as a yellow oil.Preparation 50 Chiral separation of tert-butyl indolin-3-ylcarbamate

[0102] The compound tert-butyl indolin-3-ylcarbamate (300 mg) was separated by SFC (condition: column: DAICEL CHIRALPAK AYH(250mm × 30mm,10um); mobile phase: [CO2- EtOH(0.1%NH3H2O)];B%:33%, isocratic elution mode) to give tert-butyl (S or R)-indolin-3- ylcarbamate (134.6 mg, 574.49 μmol, 44.87% yield, retention time = 0.917 min) as a white solid (Isomer 1) and tert-butyl (R or S)-indolin-3-ylcarbamate (143 mg, 610.35 μmol, 47.67% yield, retention time = 1.138 min) as a white solid (Isomer 2). EXAMPLES Example 1 N-(1-Acryloylindolin-3-yl)-2-cyanobenzo[d]thiazole-5-carboxamide (racemate)

[0103] To a solution of 2-acid (54.24 mg, 265.64 μmol) in DMF (2 mL) was added 1-(3-aminoindolin-1-yl)prop-2-en-1-one (50 mg, 266 μmol), HATU (151.51 mg, 398.46 μmol) and DIEA (171.66 mg, 1.33 mmol, 231 μL). The reaction mixture was stirred at 20 °C for 2 h. The reaction mixture was filtered and the residue was purified by prep-HPLC (neutral condition, column: Waters Xbridge BEH C18 100×30mm×10um; mobile phase: [H2O(10mM NH4HCO3)-ACN]; gradient: 25%-55% B over 8.0 min) to obtain the title compound (9.53 mg, 25.09 μmol, 10%) as a white solid.

[0104] ESI-MS (m / z) = 375 (M+H)

[0105] MW: 374.42

[0106] The compounds in the following table were prepared essentially as described in Example 1. Ex. ESI-MS No. Chemical name Structure (m / z) MW (M+H) N-(1-Acryloylindolin-3-yl)-2- 2 phenoxybenzo[d]thiazole-5- 442 441.51 carboxamide N-(1-Acryloylindolin-3-yl)-2- 3 (methylsulfonyl)benzo[d]thiaz 448 427.49 ole-5-carboxamide N-(1-Acryloyl-2,3-dihydro- 4 1H-pyrrolo[3,2-b]pyridin-3- yl)-2-cyanobenzo[d]thiazole- 390 375.41 5-carboxamide N-(1-Acryloyl-2,3-dihydro- 5 1H-pyrrolo[2,3-b]pyridin-3- yl)-2-cyanobenzo[d]thiazole- 376 375.41 5-carboxamide N-(1-Acryloyl-4- 6 aminoindolin-3-yl)-2- cyanobenzo[d]thiazole-5- 390 389.43 carboxamideN-(1-Acryloyl-4- 7 methoxyindolin-3-yl)-2- cyanobenzo[d]thiazole-5- 405 404.44 carboxamide N-(1-Acryloyl-7- 8 formamidoindolin-3-yl)-2- cyanobenzo[d]thiazole-5- 418 417.44 carboxamide N-(1-Acryloylindolin-3-yl)-2- c 397.2 9 yanobenzo[d]thiazole-5- carboxamide 374.42 (M + Na) (isomer 1) N-(1-Acryloylindolin-3-yl)-2- cyanobenzo[d] 397.2 10 thiazole-5- carboxamide 374.42 (M + Na) (isomer 2) Example 11 1-(1-Acryloylindolin-3-yl)-3-(2-cyanobenzo[d]thiazol-5-yl)urea

[0107] To a solution of 5-(100 mg, 571 μmol) in DCM (4 5 mL) was added bis(trichloromethyl)carbonate (84.68 mg, 285.37 μmol) and TEA (144.38 mg, 1.43mmol, 199 μL) at 0 °C and the mixture was stirred at 30 °C for 1 h, then 1-(3-aminoindolin-1-yl)prop- 2-en-1-one (107.43 mg, 570.74 μmol) in DCM (1mL) was added to the mixture and the mixture at 20 °C for 12 h. The residue was diluted with H2O (10 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were dried over Na2SO4, filtered and the filtrate was concentrated to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, 0-35% EtOAc / Petroleum ether gradient @ 200 mL / min) to give the title compound (86.40 mg, 219.20 μmol, 38%) as a white solid.

[0108] ESI-MS (m / z) = 486 (M+H).

[0109] MW: 389.43 Example 12 6-(((1-Acryloylindolin-3-yl)amino)methyl)benzo[d]thiazole-2-carbonitrile

[0110] To a solution of 6-(56.04 mg, 297.77 μmol) in MeOH (2 mL) was added 1-(3-aminoindolin-1-yl)prop-2-en-1-one (60 mg, 199 μmol, TFA salt) and adjusted pH=5-6 with AcOH. The mixture was stirred at 25 °C for 1 h. Then NaBH3CN (37.42 mg, 595.54 μmol) was added to the mixture. The mixture was stirred at 25 °C for 12 h. The reaction mixture was filtered and the residue was purified by prep-HPLC (neutral condition, column: Waters Xbridge BEH C18100×30mm×10um; mobile phase: [H2O (10mM NH4HCO3)-ACN]; gradient: 35%-65% B over 8.0 min) to give the title compound (11.11 mg, 30.53 μmol, 15% yield) as a white solid.

[0111] ESI-MS (m / z) = 383 (M+Na)

[0112] MW: 360.44 Biological Assays CK Target Engagement Assay CKB Protein Expression and purification

[0113] The N-terminal His tag construct of human CKB (residues 1 – 381) was overexpressed in Escherichia coli BL21 (DE3) and purified using affinity chromatography and size-exclusion chromatography. Briefly, cells were grown at 37oC in TB medium in the presence of 50 µg / mL kanamycin to an optical density of 0.8, cooled to 17oC, collected by centrifugation and stored at -80oC. Cell pellets were lysed in buffer A (25 mM HEPES, pH 7.5, 500 mM NaCl, 7 mM mercaptoethanol and 20 mM imidazole) using a Microfluidizer (Microfluidics) and the resulting lysate was centrifuged at 30,000 g for 40 min. Ni-NTA beads (Qiagen) were mixed with cleared lysate for 30 min and washed with buffer A. Beads were transferred to a fast protein liquid 5 chromatography-compatible column, and the bound protein was washed further with buffer A for 10 column volumes and eluted with buffer B (25 mM HEPES, ph 7.5, 500 mM NaCl, 7 mM mercaptoethanol, and 400 mM imidazole). The eluted samples were concentrated and purified further using a Superdex 20016 / 600 column (Cytvia) in buffer C (20 mM HEPES, ph 7.5, 200 mM Nacl, 5% glycerol, 1 mm dithiothreitol, and 0.5 mM TCEP). Fractions containing CKB were concentrated 10 to 50 mg / mL and stored at -80oC. Intact protein mass spectrometry

[0114] 1 μL of test compound (100x stock) was added to 50 μL of 800 nM human recombinant CKB and incubated for 2 h at 37 °C. The sample was then subjected to LC-MS analysis using a PLRP- S 1000A, 2.1 × 50 mm, 5 µm (Agilent) column on a Q-Exactive HF-X. The buffers used for 15 separation were 2% formic Acid in water (Buffer A) and 2% Formic Acid in acetonitrile (Buffer B) which was run at 0.3 mL / min at 60 °C. The gradient was 15% B for 0.5 mins, followed up by ramping up to 95% over 4.5 mins. Positive ion modes were collected with full scan analysis over m / z 900– 2600 m / z at 7,500 resolution, 1e5 AGC, 25 ms maximum ion accumulation time and 60 eV in-source CID. Data processing was conducted using Thermo Biopharma Finder software version 4.1 (Thermo 20 Fisher Scientific). Masses were identified demonstrating one (single), two (double), or three (triple) molecules of test compound were bound to one CKB protein molecule.

[0115] Exemplified compounds were tested essentially as described above and exhibited the following binding data.EXAMPLE MOLECULES BOUND 1 double 2 double 3 triple 4 double 5 double 6 double 7 double 8 double 9 double 10 double 11 double 12 double

[0116] These data demonstrate that the exemplified compounds label CKB. CK Inhibition Assay

[0117] Recombinant CKB was diluted to 250 nM using 50 mM TRIS, 100 μM TCEP, pH 9, and 19 μL was added to each well of a 384 well plate.1ul of compound dissolved in DMSO was added 5 to each well, resulting in 12 different concentrations ranging between 125uM to 0.03nM [each concentration was serially diluted by 1 / 4] then incubated for 2 h at 37 °C. Next, samples were diluted into assay buffer with the following final concentrations: 100 nM CKB, 40 μM TCEP, 3 mM ATP, 1.2 mM PEP, 12 mM MgCl2, 210 μM NADH, 60 mM KCl, 50 mM TRIS, 9 units / mL Pyruvate Kinase, 4.2 units / mL L-Lactate Dehydrogenase, and 6 mM creatine at pH 9. The absorbance was 10 measured at a wavelength of 340 nm over 30 mins. Measured absorbance over time in each well was then subjected to analyze the linear regression. And the slope measured from each well based on the linear regression analysis were used for calculating IC50 by PRISM software based on the following equation: Y=Bottom + (Top-Bottom) / (1+10^((LogIC50-X)*HillSlope)) 15 ^ X: Log of dose or concentration ^ Y: Response ^ Top and Bottom: Plateaus in same units as Y, Bottom-Y min, Top- Y max. ^ LogIC50: same log units as X- Valus of X at Ymid^ HillSlope: Slope factor or Hill slope- SIGN(YatMAX-YatMin)

[0118] Exemplified compounds were tested essentially as described above and exhibited the following IC50 for CK: EXAMPLE CK IC50 (µM) 1 6.55 2 n.d. 3 n.d. 4 6.8 5 n.d. 6 18.4 7 11.94 8 12.22 9 7.87 10 13.46 11 n.d. 12 n.d. n.d. = not determined 5

[0119] These data demonstrate that the compounds of Formula I are inhibitors of CK. AML and A549 Inhibition Assay

[0120] UCSD-AML1 or A549 cells were diluted to 20,000 cell / ml and then 45 μL of this suspension was introduced into every well of a 384-well plate (1125 cell / well). Following a 24-hour incubation period, the compound was first dissolved in DMSO. Subsequently, 2.5 μL of the dissolved compound 10 was mixed with 122.5 μL of culture media. Each concentration was serially diluted by 1 / 2. Then, 5 μL of the culture media, containing the compounds, was added to each well, resulting in 10 different concentrations ranging from 20 μM to 40 nM. After 3 days of incubation, 10 μL of CellTiter-Glo Luminescent Cell Viability Assay was added to each well incubated at RT for 10 min. Luminescence is measured using plate reader. All experiments are performed in 8 biological replicates per 15 condition / titrated concentration. All the data analysis was performed using PRISM. Each luminescent value was normalized to present result as percentages by averaging the subcolumns and normalizing the means. Then LD50 was calculated based on the following equation: Y=100 / (1+10^((LogLD50-X)*HillSlope)), ^ X: log of dose or concentration,^ Y: Normalized response, 100% down to 0%, ^ LogLD50: Same log units as X -Value of X at Ymid ^ HillSlope: Slope factor or Hill slope- SIGN(YatMAX-YatMin)

[0121] The exemplified compounds of Formula I were tested essentially as described above and 5 data from these assays are summarized in the following table: EXAMPLE AML IC50 (µM) A549 IC50 (µM) A549 / AML** 1 0.76 8.22 10.82 2 2.6 9.74 3.75 3 2.85 25* 8.77 4 0.54 7.77 14.39 5 0.75 2.55 3.40 6 0.73 10.77 14.75 7 0.57 7.64 13.40 8 0.67 8.01 11.96 9 0.63 8.58 13.62 10 0.37 7.84 21.19 11 1.01 17.85 17.67 12 1.45 5.21 3.59 *IC50was greater than 20 µM. A value of 25 µM was arbitrarily assigned to allow the calculation of the A549 / AML ratio. **CK is essential for AML cell viability but A549 cells are a representative non-CK sensitive cell type. The ratio of A549 IC50to AML IC50(A549 / AML) greater than 1 for a compound of Formula 10 I demonstrates engagement of the CK protein.

Claims

CLAIMS We claim:

1. A compound of Formula I, or a pharmaceutically acceptable salt thereof: where:A is phenyl or pyridinyl; R1is an optional substituent that, when present, is bonded to a ring carbon atom and is selected from the group consisting of C1 – C3 alkoxy, cyclopropoxy, amino, -NHCHO, and -NHC(O)CH3; X is -C(O)-, -C(O)NH-, or -CH2-; ;cycloalkyl).

2. A compound of Claim 1 or a pharmaceutically acceptable salt thereof wherein A is pyridinyl.

3. A compound of Claim 2 or a pharmaceutically acceptable salt thereof wherein A is pyridinyl and R1is absent.

4. A compound of Claim 1 or a pharmaceutically acceptable salt thereof wherein A is phenyl.

5. A compound of Claim 4 where A is phenyl and R1is absent.

6. A compound of any of Claims 1 – 5 or a pharmaceutically acceptable salt thereof where X is -C(O)- or -C(O)NH-.

7. A compound of any of Claims 1 – 6 or a pharmaceutically acceptable salt thereof where Y is .any of Claims 1 – 7 or a pharmaceutically acceptable salt thereof where R2is -CN.

9. A pharmaceutical composition comprising a compound of any of Claims 1 – 8 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

10. A compound of any of Claims 1 – 8 or a pharmaceutically acceptable salt thereof for use in therapy.

11. A method for treating a disorder susceptible to inhibition of CK comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any of Claims 1 – 8 or a pharmaceutically acceptable salt thereof.

12. A method of Claim 11 where the disorder is cancer.

13. A method of Claim 12 where the disorder is liver cancer, pancreatic cancer, lung cancer, colon cancer, and hematological cancers, including myelogenous leukemia, myeloid leukemia (including acute myeloid leukemia), myelodysplastic syndrome, lymphoblastic leukemia (including acute lymphoblastic leukemia), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), high risk CLL, follicular lymphoma, diffuse large B-cell lymphoma (including DLBCL or ABC-DLBCL), mantle cell lymphoma (MCL), Waldenstrom’s macroglobulinemia (WM), multiple myeloma, marginal zone lymphoma (MZL), Burkitt’s lymphoma, non-Burkitt high grade B-cell lymphoma, extranodal marginal zone B-cell lymphoma, transformed high grade B-cell lymphoma(HGBL), lymphoplasmacytic lymphoma (LPL), central nervous system lymphoma (CNSL), or MALT lymphoma.

14. A method of Claim 13 where the disorder is acute myeloid leukemia. 5 15. A compound of any of Claims 1 - 8 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of disorders susceptible to inhibition of CK.

16. The use of Claim 15 where the disorder is cancer.

17. The use of Claim 16 where the disorder is liver cancer, pancreatic cancer, lung cancer, colon cancer, and hematological cancers, including myelogenous leukemia, myeloid leukemia (including acute myeloid leukemia), myelodysplastic syndrome, lymphoblastic leukemia (including acute lymphoblastic leukemia), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), high risk CLL, follicular lymphoma, diffuse large B-cell lymphoma (including DLBCL or ABC-DLBCL), mantle cell lymphoma (MCL), Waldenstrom’s macroglobulinemia (WM), multiple myeloma, marginal zone lymphoma (MZL), Burkitt’s lymphoma, non-Burkitt high grade B-cell lymphoma, extranodal marginal zone B-cell lymphoma, transformed high grade B-cell lymphoma (HGBL), lymphoplasmacytic lymphoma (LPL), central nervous system lymphoma (CNSL), or MALT lymphoma.

18. The use of Claim 17 where the disorder is acute myeloid leukemia.

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