CDK6 / DYRK2 dual-target inhibitor, method for manufacturing the same, and use of the same

Abstract

The present invention discloses a compound represented by the following chemical formula (I) or a pharmaceutically acceptable salt thereof. The present invention also discloses a method for preparing said compound and the use of said compound in the prevention and / or treatment of cancer or tumor-related diseases, particularly breast cancer, prostate cancer, lung cancer, multiple myeloma, leukemia, gastric cancer, ovarian cancer, colorectal cancer, liver cancer, pancreatic cancer, human glioma, etc. The compound of the present invention is expected to be developed as a next-generation anticancer agent.

Description

Technology Field

[0001] The present invention relates to the field of pharmaceutical chemistry, particularly to CDK6 / DYRK2 dual-target inhibitors, methods for manufacturing the same, and uses the same. Background Technology

[0002] Cyclin-dependent kinase 6 (CDK6) is a serine / tyrosine kinase that regulates the transition from G1 to S in the cell cycle. During the early G1 phase of the cell cycle, cyclin D binds to and activates CDK6, and the resulting cyclin D-CDK6 complex promotes the phosphorylation of retinoblastoma protein (Rb). The phosphorylation of Rb leads to the release of transcription factor E2F, which accelerates the progression of the cell cycle from G1 to S. The upregulation of the proto-oncogene CDK6 induces accelerated progression of the cell cycle from G1 to S, thereby leading to accelerated cell cycle and proliferation. Uncontrolled cell proliferation is a key characteristic of cancer. Therefore, inhibition of CDK6 can exhibit antiproliferative and anticancer effects by slowing the transition of the cell cycle from the G1 to S phase. However, currently marketed CDK6 inhibitors—palbociclib, ribociclib, and abemaciclib—are highly toxic and have developed resistance.

[0003] Bispecific tyrosine phosphorylation-regulating kinases (DYRK) and CDKs belong to the CMGC family and play important regulatory roles in the cell cycle and cell proliferation. DYRK2 regulates the cell cycle-dependent phosphorylation of Rpt3-T25 and promotes the degradation of CDK inhibitors such as p21 and p27, as well as accelerating the progression of the cell cycle from G1 to S. Inhibition of DYRK2 can also slow the cell cycle transition from the G1 phase to the S phase, leading to antiproliferative and anticancer effects. To date, only a small number of DYRK2 inhibitors have been reported. The acridine compound LDN192960 was originally identified as a rhizine kinase inhibitor with some therapeutic effects in triple-negative breast cancer and multiple myeloma. Curcumin, another drug confirmed to act on DYRK2 and DYRK3, may exhibit specific anti-multiple myeloma effects when used in combination with carfilzomib. However, the anticancer activity and target selectivity of existing DYRK2 inhibitors still need to be optimized, and in particular, their drug formation characteristics need to be further improved.

[0004] Targeted drugs exhibit potent efficacy and excellent safety. However, due to the complexity and integrity of cancer, when a single-target drug inhibits one pathway of the cancer, related pathways are activated to compensate for the inhibited one, leading to drug resistance. The problem to be solved

[0005] To address the problem of drug resistance caused by existing drugs in single-target therapies, the present invention provides a compound or a pharmaceutically acceptable salt thereof that can be simultaneously targeted to CDK6 and DYRK2 (wherein the compound is a CDK6 / DYRK2 dual-target inhibitor) by utilizing the synergistic effect of CDK6 and DYRK2, and enhances the anticancer activity of the compound and reduces drug resistance easily caused by CDK6 single-target drugs by inhibiting DYRK2 and blocking the compensation pathway of CDK6. The present invention also provides a specific method for manufacturing a compound and a drug for preventing and / or treating cancer or tumor-related diseases, particularly including breast cancer, prostate cancer, lung cancer, multiple myeloma, leukemia, gastric cancer, ovarian cancer, colon cancer, liver cancer, pancreatic cancer, human glioma, etc., and is expected to be developed as a next-generation anticancer agent. means of solving the problem

[0006] The present invention relates to a compound represented by the formula (I) or a pharmaceutically acceptable salt thereof:

[0007]

[0008] Here, X is O, (CH2) n , C(O), NH or S(O)2, and n is 0 or 1;

[0009] R1 is hydrogen, deuterium, halogen, hydroxyl group, mercapto group, cyano group, nitro group, C1-C8 alkyl group, halo-C1-C8 alkyl group, C1-C8 alkoxy group, C3-C8 cycloalkyl group, C6-C 10 Aryl group, C3-C 10 Heteroaryl group, C4-C8 heterocyclic group, -C 0-8 Selected from -NR4R5;

[0010] R2 is selected from hydrogen, deuterium, halogen, hydroxyl group, mercapto group, cyano group, nitro group, C1-C8 alkyl group, and C3-C8 cycloalkyl group;

[0011] R3 is hydrogen, deuterium, halogen, C1-C8 alkyl group, halo-C1-C8 alkyl group, C1-C8 alkoxy group, C3-C8 cycloalkyl group, -C 0-8 -S(O)2R6, -C 0-8 Selected from -C(O)OR7;

[0012] R4 and R5 are each independently selected from hydrogen, deuterium, C1-C8 alkyl groups, halo-C1-C8 alkyl groups, C1-C8 alkoxy groups, and C3-C8 cycloalkyl groups;

[0013] R6 and R7 are each independently selected from hydrogen, C1-C8 alkyl groups, halo-C1-C8 alkyl groups, and C3-C8 cycloalkyl groups;

[0014] Preferably,

[0015] The above X is C(O) or (CH2) n and; n is 0 or 1 and;

[0016] The above R1 is hydrogen, a C1-C8 alkyl group, or -C 0-8 -NR4R5, where R4 and R5 are hydrogen and C1-C8 alkyl groups. or selected from C3-C8 cycloalkyl groups;

[0017] The above R2 is hydrogen or halogen and ;

[0018] The above R3 is hydrogen, C1-C8 alkyl group, -C 0-8 -S(O)2R6, or -C 0-8 -C(O)OR7 and R6, R7 are selected from C1-C8 alkyl groups, and

[0019] Preferably,

[0020] The above X is C(O) or (CH2) n and; n is 0 or 1 and;

[0021] The above R1 is hydrogen, C1-C3 alkyl group or selected from -NR4R5, where R4 and R5 are selected from hydrogen, C1-C3 alkyl groups, cyclopentane, or cyclohexane;

[0022] The above R2 is hydrogen or F;

[0023] The above R3 is hydrogen or a C1-C4 alkyl group, -S(O)2R6 or -C(O)OR7, where R6 and R7 are selected from C1-C4 alkyl groups;

[0024] Preferably,

[0025] The above X is C(O) or (CH2) n and; n is 0 or 1 and;

[0026] The above R1 is selected from hydrogen, a methyl group, or -NR4R5, wherein R4 and R5 are selected from hydrogen, a methyl group, an ethyl group, or cyclopentane;

[0027] R2 is F and;

[0028] R3 is selected from hydrogen, ethyl group, isopropyl group, -S(O)2R6 or -C(O)OR7, R6 is a methyl group, and R7 is selected from a tert-butyl group or an ethyl group.

[0029] Preferably,

[0030] The above X is C(O) or (CH2) n Selected from; n is 0 or 1 and;

[0031] The above R1 is selected from hydrogen, a methyl group, or -NR4R5, wherein R4 and R5 are selected from hydrogen, a methyl group, an ethyl group, or cyclopentane;

[0032] The above R2 is F and ;

[0033] The above R3 is selected from hydrogen, ethyl group, isopropyl group, or -C(O)OR7, and R7 is selected from tert-butyl group.

[0034] Preferably,

[0035] The above X is (CH2) n or selected from C(O) , n is 0 or 1 and;

[0036] The above R1 is selected from hydrogen, a methyl group, or -NR4R5 and , Here, R4 is selected from hydrogen, methyl group, and ethyl group, and R5 is selected from hydrogen, methyl group, ethyl group, or cyclopentane;

[0037] The above R2 is F;

[0038] The above R3 is It is selected from hydrogen, ethyl group, isopropyl group or -C(O)OR7, and R7 is selected from tert-butyl group.

[0039] Preferably, the compounds of the present application are selected from I-1 to I-53:

[0040]

[0041]

[0042]

[0043]

[0044]

[0045] Preferably:

[0046] The above X is (CH2) n or C(O), n is selected from 0 or 1;

[0047] The above R1 is Selected from hydrogen, methyl group, or -NR4R5, where R4 and R5 are each independently selected from hydrogen, methyl group, and ethyl group; or R4 is h is, and R5 is It is cyclopentane;

[0048] The above R2 is F;

[0049] The above R3 is It is selected from hydrogen, ethyl group, and isopropyl group.

[0050] The above pharmaceutically acceptable salt is an acidic addition salt of a compound of formula (I), wherein the salt-forming acid comprises an inorganic acid and an organic acid, the inorganic acid comprises hydrochloric acid, sulfuric acid, phosphoric acid and methanesulfonic acid, and the organic acid comprises acetic acid, trichloroacetic acid, propionic acid, butyric acid, maleic acid, p-toluenesulfonic acid, malic acid, malonic acid, cinnamic acid, citric acid, fumaric acid, camphor, digluconic acid, aspartic acid and tartaric acid.

[0051] Preferably, the pharmaceutically acceptable salt of the present invention is a hydrochloride salt.

[0052] The present invention relates to a method for preparing a compound of the following formula (I): preparing compound (I) from compound (A) and compound (B) through a coupling reaction under the action of a palladium catalyst:

[0053]

[0054] Here,

[0055] X is O, (CH2) n, Selected from C(O), NH or S(O)2, where n is 0 or 1;

[0056] R1 is hydrogen, deuterium, halogen, hydroxyl group, mercapto group, cyano group, nitro group, C1-C8 alkyl group, halo-C1-C8 alkyl group, C1-C8 alkoxy group, C1-C8 alkoxy group, C3-C8 cycloalkyl group, C6-C 10 Aryl group, C3-C 10 Heteroaryl group, C4-C8 heterocyclic group, -C 0-8 Selected from -NR4R5,

[0057] R2 is selected from hydrogen, deuterium, halogen, hydroxyl group, mercapto group, cyano group, nitro group, C1-C8 alkyl group, and C3-C8 cycloalkyl group;

[0058] R3 is hydrogen, deuterium, C1-C8 alkyl group, halo-C1-C8 alkyl group , C1-C8 alkoxy group, C3-C8 cycloalkyl group, -C 0-8-S(O)2R6, -C 0-8 Selected from -C(O)OR7;

[0059] R4 and R5 are each independently selected from hydrogen, deuterium, C1-C8 alkyl groups, halo-C1-C8 alkyl groups, C1-C8 alkoxy groups, and C3-C8 cycloalkoxy groups;

[0060] R6 and R7 are each independently selected from hydrogen, C1-C8 alkyl groups, halo-C1-C8 alkyl groups, and C3-C8 cycloalkyl groups.

[0061] Preferably, the reaction is carried out under an argon protective atmosphere; the reaction temperature is 95-105°C, and preferably the reaction temperature is 100°C.

[0062] The present invention also discloses a pharmaceutical composition comprising a compound of the aforementioned formula (I), a pharmaceutically acceptable salt thereof, or an isomer thereof, and a pharmaceutically acceptable carrier.

[0063] Pharmaceutically acceptable carriers refer to excipients or diluents that do not cause significant irritation to the organism and do not interfere with the biological activity and properties of the administered compound.

[0064] The present invention relates to the use of a compound or a pharmaceutically acceptable salt thereof in the preparation of a CDK6 / DYRK2 dual-target inhibitor.

[0065] CDK6 / DYRK2 dual-target inhibitor drugs can be used to treat cancer or tumor-related diseases.

[0066] The present invention relates to the use of a compound or a pharmaceutically acceptable salt thereof in the manufacture of a drug for preventing and / or treating cancer or tumor-related diseases. Cancer or tumor-related diseases include breast cancer, prostate cancer, lung cancer, multiple myeloma, leukemia, gastric cancer, ovarian cancer, colorectal cancer, liver cancer, pancreatic cancer, and human glioma.

[0067] The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof having CDK6 / DYRK2 dual-target inhibitory activity and exhibiting a therapeutic effect against cell malignant proliferative tumors.

[0068] Unless otherwise specified, the terms used in this invention have the following meanings.

[0069] The term "alkyl group" refers to a linear or branched saturated hydrocarbon group having a number of carbon atoms.

[0070] Term "C 1- "C8 alkyl group" refers to a linear or branched saturated hydrocarbon group having 1 to 8 carbon atoms. C1-C8 alkyl groups include, but are not limited to, methyl groups, ethyl groups, n-propyl groups, isopropyl groups, n-butyl groups, isobutyl groups, tert-butyl groups, n-pentyl groups, isopentyl groups, neopentyl groups, n-hexyl groups, isohexyl groups, 2,2-dimethylbutyl groups, and 2,3-dimethylbutyl groups. The term "C1-C3 alkyl group" refers to a linear or branched saturated hydrocarbon group having 1 to 3 carbon atoms.

[0071] The term "alkoxy group" means an O-alkyl group. The term "C1-C8 alkoxy group" refers to a group having an O-C1-C8 alkyl group.

[0072] "C(O)" means "-C(O)-", specifically a carbonyl group. The term "halogen" is fluorine, chlorine, bromine, or iodine. Preferably, it is fluorine, chlorine, or bromine.

[0073] The term "halo-alkyl group" means an alkyl group having at least one (including one) halogen substituent.

[0074] The term "cycloalkyl group" refers to a saturated monocyclic or polycyclic ring structure composed of carbon atoms.

[0075] Term "C 3-"C8 cycloalkyl group" refers to a saturated monocyclic or polycyclic ring structure having a total of 3 to 8 atoms. C3-C6 cycloalkyl groups include, but are not limited to, cyclopropyl groups, cyclobutyl groups, cyclopentyl groups, and cyclohexyl groups.

[0076] The term "cycloalkenyl" refers to a monocyclic or polycyclic alkyl substituent having at least one cyclic carbon-carbon double bond.

[0077] The term "C3-C8 cycloalkenyl" refers to a cycloalkenyl having 3 to 8 carbon atoms. C3-C8 cycloalkenyls include, but are not limited to, cyclopentenyl groups and cyclobutenyl groups.

[0078] The term "C2-C8 alkenyl" refers to a linear or branched hydrocarbon group having one or more carbon-carbon double bonds and 2 to 8 carbon atoms.

[0079] Term "C 2- "C8 alkynyl" refers to a linear or branched hydrocarbon group having one or more carbon-carbon triple bonds and 2 to 8 carbon atoms.

[0080] Term "C 6- C 10 "Aryl group" means a monocyclic or fused polycyclic group having a fully conjugated π-electron system consisting of 6 to 10 carbon atoms, and typically includes, but is not limited to, phenyl groups and naphthyl groups.

[0081] The term "heteroaryl group" means a monocyclic or fused ring having 1, 2, 3, or 4 cyclic heteroatoms selected from the group consisting of N, O, or S, the remainder of the cyclic atoms being C, and additionally having a fully conjugated π-electron system. The term "C3-C 10 "Heteroaryl group" refers to a heteroaryl group having 3 to 10 carbon atoms in a ring. C3-C 10Heteroaryl groups include, but are not limited to, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyrimidine, and pyridine.

[0082] The term "heterocyclic group" refers to a monocyclic or fused ring group having one or more heteroatoms of N, O, or S as a heterocycloalkyl group. The term "C4-C8 heterocyclic group" refers to a heterocyclic group having 4 to 8 carbon atoms in the ring. C4-C8 heterocyclic groups include, but are not limited to, piperazino groups, morpholino groups, piperidino groups, pyrrolidino groups, etc. Effects of the invention

[0083] Compared to the prior art, the present invention discloses a novel compound represented by general formula (I) capable of simultaneously inhibiting multiple pathways of cancer, which exhibits excellent therapeutic effects, low toxicity, good drug metabolic characteristics, and is resistant to developing drug resistance, and can be used in the manufacture of drugs for treating cancer or tumor-related diseases. The present invention also discloses a method for manufacturing a compound of formula (I). Brief explanation of the drawing

[0084] Figure 1 shows the change in body weight of a mouse in the acute toxicity analysis of the present invention. Figure 2 is a graph showing the HE staining results in the acute toxicity analysis method of the present invention. Figure 3 is a graph of the results for the prostate cancer tumor volume according to the present invention. Specific details for implementing the invention

[0085] The present application is described in detail below with reference to specific symbols.

[0086] I. Synthesis of Intermediate Reactants

[0087] Reactant (A) and reactant (B) can be purchased directly or developed independently, and costs can be drastically reduced by developing them independently. Specific manufacturing methods for independently developed reactant (A) and reactant (B) are as follows.

[0088] (1) Synthesis of 6-(2-chloro-5-fluoropyrimidine-4-yl)benzothiazole (A-1)

[0089]

[0090] Step 1. Synthesis of 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzothiazole: 6-bromobenzothiazole (0.43 g, 2.0 mmol) was dissolved in DMF (10 mL). Subsequently, pinacol borate (0.53 g, 2.1 mmol), Pd(dppf)Cl2 (22 mg, 0.06 mmol), and potassium acetate (0.59 g, 6.0 mmol) were added. The reaction was carried out for 24 hours at 80°C with argon replaced three times. The mixture was cooled, filtered, and concentrated, and purified by flash silica gel column chromatography to obtain compound 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzothiazole (0.47 g, 90% yield).

[0091] 1 HNMR (300 MHz, CDCl3) δ 9.07 (s, 1H), 8.46 (s, 1H), 8.14 (d, J = 8.2 Hz, 1H), 7.94 (dd, J = 8.2, 1.1 Hz, 1H), 1.38 (s, 12H).

[0092]

[0093] Step 2. Synthesis of 6-(2-chloro-5-fluoropyrimidine-4-yl)benzothiazole (A-1): Compound 2,4-dichloro-5-fluoropyrimidine (0.23 g, 1.4 mmol) was weighed and placed in a 250 mL three-necked flask. Then, Pd(PPh3)2Cl2 (21 mg, 0.03 mmol), sodium carbonate (0.27 g, 2.5 mmol), glyme (10 mL), and H2O (0.25 mL) were added. The reaction was heated to 80°C with argon replaced three times. Compound 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzothiazole (0.26 g, 1.0 mmol) was dissolved in glyme (5 mL), added dropwise to a 3-necked flask, and reacted for 16 hours. The mixture was cooled, filtered, and concentrated, and purified by flash silica gel column chromatography to obtain compound 6-(2-chloro-5-fluoropyrimidine-4-yl)benzothiazole (0.22 g, 82% yield).

[0094] 1 HNMR (300 MHz, CDCl3) δ 9.17 (s, 1H), 8.84 (d, J = 1.7 Hz, 1H), 8.58 (d, J = 3.1 Hz, 1H), 8.37 - 8.24 (m, 2H).

[0095] (2) Synthesis of 6-(2-chloro-5-fluoropyrimidine-4-yl)-2-methylbenzothiazole(A-2)

[0096]

[0097] Referring to the synthesis of compound (A-1), the yields were 90% and 84%, respectively.

[0098] 1 H NMR (400 MHz, CDCl3) δ 8.70 (d, J = 1.9 Hz, 1H), 8.55 (d, J = 3.1 Hz, 1H), 8.28 - 8.25 (m, 1H), 8.07 (d, J = 8.6 Hz, 1H), 2.90 (s, 3H).

[0099] (3) Synthesis of 6-(2-chloro-5-fluoropyrimidine-4-yl)-N-cyclopentylbenzothiazole-2-amine (A-3)

[0100]

[0101] Step 1. Synthesis of 6-Bromo-N-Cyclopentylbenzothiazole-2-amine: 6-Bromo-2-chlorobenzothiazole (0.50 g, 2.0 mmol) was dissolved in DMSO (10 mL), and cyclopentylamine (0.19 g, 2.2 mmol) and N-ethyldiisopropylamine (0.39 g, 3.0 mmol) were added. The argon was replaced three times during the reaction, and the mixture was heated to 80°C and reacted for 12 hours. The mixture was cooled, filtered, concentrated, and purified by flash silica gel column chromatography to obtain the compound 6-Bromo-N-Cyclopentylbenzothiazole-2-amine (0.53 g, 90% yield).

[0102] 1 H NMR (400 MHz, CDCl3) δ 7.68 (d, J = 1.7 Hz, 1H), 7.38 - 7.33 (m, 2H), 6.28 (s, 1H), 3.98 - 3.93 (m, 1H), 2.14 - 2.04 (m, 2H), 1.72 - 1.54 (m, 6H).

[0103]

[0104] Step 2. Synthesis of 6-(2-chloro-5-fluoropyrimidine-4-yl)-N-cyclopentylbenzothiazole-2-amine (A-3): Referring to the synthesis of compound (A-1), the yields were 88% and 83%, respectively.

[0105] 1 H NMR (400 MHz, CDCl3) δ 8.49 (d, J = 1.9 Hz, 1H), 8.46 (d, J = 3.5 Hz, 1H), 8.17 - 8.14 (m, 1H), 7.58 (d, J= 8.6 Hz, 1H), 5.88 (s, 1H), 4.13 - 4.07 (m, 1H), 2.19 - 2.11 (m, 2H), 1.77 - 1.61 (m, 6H).

[0106] (4) Synthesis of 6-(2-chloro-5-fluoropyrimidine-4-yl)-N,N-dimethylbenzothiazole-2-amine (A-4)

[0107]

[0108] Step 1. Synthesis of 6-Bromo-N,N-Dimethylbenzothiazole-2-amine: 4-Bromo-2-Iodoaniline (0.60 g, 2.0 mmol), sodium dimethyl dithiocarbamate dihydrate (0.72 g, 4.0 mmol), copper acetate (0.36 g, 2.0 mmol), and potassium carbonate (0.55 g, 4.0 mmol) were weighed, dissolved in DMF (10 mL), and reacted at 120°C for 6 hours. The mixture was cooled, filtered, concentrated, and purified by flash silica gel column chromatography to obtain the compound 6-Bromo-N,N-Dimethylbenzothiazole-2-amine (0.44 g, 85% yield).

[0109] 1 H NMR (400 MHz, CDCl3) δ 7.69 (d, J = 1.9 Hz, 1H), 7.41 - 7.35 (m, 2H), 3.20 (s, 6H).

[0110]

[0111] Step 2. Synthesis of 6-(2-chloro-5-fluoropyrimidine-4-yl)-N,N-dimethylbenzothiazole-2-amine (A-4): Referring to the synthesis of compound (A-1), the yields were 88% and 80%.

[0112] 1 H NMR (400 MHz, CDCl3) δ 8.49 (d, J = 1.9 Hz, 1H), 8.45 (d, J = 3.6 Hz, 1H), 8.17 - 8.14 (m, 1H), 7.62 (d,J = 8.7 Hz, 1H), 3.27 (s, 6H).

[0113] (5) Synthesis of 6-(2-chloro-5-fluoropyrimidine-4-yl)-N,N-diethylbenzothiazole-2-amine (A-5)

[0114]

[0115] Step 1. Synthesis of 6-Bromo-N,N-Diethylbenzothiazole-2-amine: 4-Bromo-2-Iodoaniline (0.60 g, 2.0 mmol), sodium diethyldithiocarbamate trihydrate (0.90 g, 4.0 mmol), copper acetate (0.36 g, 2.0 mmol), and potassium carbonate (0.55 g, 4.0 mmol) were weighed, dissolved in DMF (10 mL), and reacted at 120°C for 6 hours. The mixture was cooled, filtered, concentrated, and purified by flash silica gel column chromatography to obtain the compound 6-Bromo-N,N-Dimethylbenzothiazole-2-amine (0.46 g, yield 80%).

[0116]

[0117] Step 2. Synthesis of 6-(2-chloro-5-fluoropyrimidine-4-yl)-N,N-diethylbenzothiazole-2-amine (A-5): Referring to the synthesis of compound (A-1), the yields were 90% and 82%. 1 H NMR (300 MHz, CDCl3) δ 8.44 (dd, J = 8.8, 2.7 Hz, 2H), 8.13 (d, J = 8.6 Hz, 1H), 7.58 (d, J = 8.7 Hz, 1H), 3.61 (q, J = 7.2 Hz, 4H), 1.32 (t, J = 7.2 Hz, 6H).

[0118] (6) Synthesis of (6-aminopyridine-3-yl)(4-ethylpiperazine-1-yl)ketone (B-1)

[0119]

[0120] 6-aminonicotinic acid (0.28 g, 2.0 mmol) and N,N'-carbonyldiimidazole (0.39 g, 2.4 mmol) were weighed, dissolved in DMF (5 mL), and reacted at 70°C for 10 minutes. N-ethylpiperazine (0.46 g, 4.0 mmol) was added for 1 hour at room temperature. The reaction was carried out overnight at room temperature, concentrated, and purified by flash silica gel column chromatography to obtain the compound (6-aminopyridine-3-yl)(4-ethylpiperazine-1-yl)ketone (0.40 g, 85% yield).

[0121] 1 H NMR (300 MHz, CDCl3) δ 8.19 - 8.17 (m, 1H), 7.57 - 7.54 (m, 1H), 6.51 - 6.48 (m, 0.9 Hz, 1H), 4.79 (s, 2H), 3.73 - 3.60 (m, 4H), 2.49 - 2.42 (m, 6H), 1.13 - 1.08 (m, 3H).

[0122] (7) Synthesis of 5-((4-ethylpiperazine-1-yl)methyl)pyridine-2-amine (B-2)

[0123]

[0124] 2-amino-5-formylpyridine (0.32 g, 2.6 mmol) and N-ethylpiperazine (0.45 g, 3.9 mmol) were dissolved in 1,2-dichloroethane (20 mL) and stirred at room temperature for 2 hours. Then, sodium triacetylborohydride (0.87 g, 4.1 mmol) was added. The reaction was stirred at room temperature for 8 hours. 1 M NaOH (30 mL) was added and the mixture was quenched. The mixture was extracted with DCM (20 mL × 3), dried and concentrated with anhydrous sodium sulfate, and then subjected to column chromatography (DCM / MeOH = 10:1) to obtain compound 5-((4-ethylpiperazine-1-yl)methyl)pyridine-2-amine (0.52 g, 91%).

[0125] 1 HNMR (300 MHz, CDCl3) d 7.94 (d,J = 2.3 Hz, 1H), 7.40 (dd, J = 8.3, 2.4 Hz, 1H), 6.46 (d, J = 8.3 Hz, 1H), 4.57 (s, 2H), 3.36 (s, 2H), 2.47-2.37 (m, 10H), 1.07 (t, J = 7.2 Hz, 3H).

[0126] (8) Synthesis of tert-butyl 4-((6-aminopyridine-3-yl)methyl)piperazine-1-carboxylate (B-3)

[0127]

[0128] Referring to the synthesis of compound (B-2), the yield was 89%.

[0129] 1 HNMR (300 MHz, CDCl3): d 7.94 (d, J = 2.3 Hz, 1H), 7.40 (dd, J = 8.4, 2.3 Hz, 1H), 6.48 (d, J = 8.4 Hz, 1H), 4.54 (s, 2H), 3.40 (t, J = 5.1 Hz, 4H), 3.36 (s, 2H), 2.35 (t, J = 5.1 Hz, 4H), 1.45 (s, 9H).

[0130] (9) Synthesis of tert-butyl 4-(6-aminonicotinoyl)piperazine-1-carboxylate (B-4)

[0131]

[0132] Referring to the synthesis of compound (B-1), the yield was 87%.

[0133] 1 H NMR (300 MHz, CDCl3) δ 8.18 (d, J = 2.2 Hz, 1H), 7.56 (dd, J = 8.5, 2.2 Hz, 1H), 6.51 (d, J= 8.5 Hz, 1H), 4.76 (s, 2H), 3.65 - 3.56 (m, 4H), 3.48 - 3.42 (m, 4H), 1.48 (s, 9H).

[0134] (10) Synthesis of tert-butyl 4-(6-aminopyridine-3-yl)piperazine-1-carboxylate (B-5)

[0135]

[0136] Step 1. Synthesis of tert-butyl 4-(6-nitropyridine-3-yl)piperazine-1-carboxylate: 5-bromo-2-nitropyridine (0.41 g, 2.0 mmol), tert-butylpiperazine-1-carboxylate (0.48 g, 2.6 mmol), and triethylamine (0.41 g, 4.0 mmol) were weighed, dissolved in DMSO (5 mL), heated to 60°C, and reacted for 18 hours. The mixture was cooled, filtered, concentrated, and purified by flash silica gel column chromatography to obtain the compound tert-butyl 4-(6-nitropyridine-3-yl)piperazine-1-carboxylate (0.49 g, 80% yield).

[0137] 1 H NMR (400 MHz, CDCl3) δ 8.17 - 8.13 (m, 2H), 7.22 (dd, J = 9.2, 3.1 Hz, 1H), 3.66 - 3.64 (m, 4H), 3.49 - 3.46 (m, 4H), 1.49 (s, 9H).

[0138]

[0139] Step 2. Synthesis of tert-butyl 4-(6-aminopyridine-3-yl)piperazine-1-carboxylate: 0.31 g, 1.0 mmol of tert-butyl 4-(6-nitropyridine-3-yl)piperazine-1-carboxylate, 0.17 g, 3.0 mmol of reduced iron powder, and 0.49 g, 9.0 mmol of ammonium chloride were weighed, dissolved in 10 mL of 70% ethanol, heated to 70°C, and reacted for 6 hours. The mixture was cooled, filtered, concentrated, and purified by flash silica gel column chromatography to obtain the compound tert-butyl 4-(6-aminopyridine-3-yl)piperazine-1-carboxylate (0.24 g, 85% yield).

[0140] 1 H NMR (300 MHz, CDCl3) δ 7.78 (d, J = 2.9 Hz, 1H), 7.17 (dd, J = 8.8, 2.9 Hz, 1H), 6.49 (d, J = 8.8 Hz, 1H), 4.19 (s, 2H), 3.59 - 3.55 (m, 4H), 2.98 - 2.94 (m, 4H), 1.48 (s, 9H).

[0141] (11) Synthesis of 5-((4-(methanesulfonyl)piperazine-1-yl)methyl)pyridine-2-amine (B-6)

[0142]

[0143] Referring to the synthesis of compound (B-2), the yield was 90%.

[0144] 1 H NMR (400 MHz, CDCl3) δ 8.10 - 7.79 (m, 1H), 7.40 (d, J = 8.3 Hz, 1H), 6.57 (s, 1H), 4.55 (s, 2H), 3.41 (s, 2H), 3.22 (t, J = 4.9 Hz, 4H), 2.77 (s, 3H), 2.53 (t, J = 5.0 Hz, 4H).

[0145] (12) Synthesis of ethyl 4-((6-aminopyridine-3-yl)methyl)piperazine-1-carboxylate (B-7)

[0146]

[0147] Referring to the synthesis of compound (B-2), the yield was 86%.

[0148] 1 H NMR (400 MHz, CDCl3) δ 7.87 (d, J = 2.2 Hz, 1H), 7.43 (dd, J = 8.5, 2.3 Hz, 1H), 6.51 (d, J = 8.5 Hz, 1H), 5.71 (s, 2H), 4.12 (q, J = 7.1 Hz, 2H), 3.46 (t, J = 5.1 Hz, 4H), 3.36 (s, 2H), 2.37 (t, J = 5.1 Hz, 4H), 1.25 (t, J = 7.1 Hz, 3H).

[0149] (13) Synthesis of 5-((4-isopropylpiperazine-1-yl)methyl)pyridine-2-amine (B-8)

[0150]

[0151] Referring to the synthesis of compound (B-2), the yield was 81%.

[0152] 1 H NMR (400 MHz, CDCl3) δ 7.88 (d, J = 2.2 Hz, 1H), 7.43 (dd, J = 8.4, 2.2 Hz, 1H), 6.48 (d, J = 8.4 Hz, 1H), 4.95 (s, 2H), 3.39 (s, 2H), 2.88 - 2.80 (m, 1H), 2.72 - 2.51 (m, 8H), 1.10 (d, J = 6.6 Hz, 6H).

[0153] (14) Synthesis of (6-aminopyridine-3-yl)(4-isopropylpiperazine-1-yl)ketone (B-9)

[0154]

[0155] Referring to the synthesis of compound (B-1), the yield was 85%.

[0156] 1 H NMR (300 MHz, CDCl3) δ 8.18 (s, 1H), 7.55 (d, J = 8.4 Hz, 1H), 6.49 (dd, J = 8.6, 2.1 Hz, 1H), 4.86 (s, 2H), 3.67 - 3.61 (m, 4H), 2.77 - 2.71 (m, 1H), 2.55 - 2.51 (m, 4H), 1.05 (d, J = 6.0 Hz, 6H).

[0157] II. Synthesis of Compounds I-1 to I-53

[0158] Example 1:

[0159] Synthesis of (6-((4-(benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)(4-ethylpiperazine-1-yl)ketone(I-1):

[0160]

[0161] Compounds 6-(2-chloro-5-fluoropyrimidine-4-yl)benzothiazole (133 mg, 0.5 mmol) and 6-aminopyridine-3-yl)(4-ethylpiperazine-1-yl)ketone (141 mg, 0.6 mmol) were dissolved in dioxane (5 mL). Subsequently, Pd2(dba)3 (23 mg, 0.025 mmol), xantphos (58 mg, 0.1 mmol), and cesium carbonate (326 mg, 1.0 mmol) were added. The reaction was carried out for 12 hours at 100°C with the argon replaced three times. The mixture was cooled, filtered, and concentrated, and column chromatography (DCM~DCM / MeOH=10:1) was performed to obtain the compound (6-((4-(benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)(4-ethylpiperazine-1-yl)ketone (88 mg, 38% yield).

[0162] 1 H NMR (300 MHz, CDCl3) δ 9.68 (s, 1H), 9.15 (s, 1H), 8.76 - 8.75 (m, 1H), 8.61 - 8.58 (m, 2H), 8.52 (dd, J = 8.7, 0.9 Hz, 1H), 8.33 - 8.26 (m, 2H), 7.86 (dd, J = 8.7, 2.3 Hz, 1H), 3.80 - 3.61 (m, 4H), 2.51 - 2.44 (m, 6H), 1.12 (t, J = 7.2 Hz, 3H).

[0163] Example 2:

[0164] Synthesis of 4-(benzothiazole-6-yl)-N-(5-((4-ethylpiperazine-1-yl)methyl)pyridine-2-yl)-5-fluoropyrimidine-2-amine(I-2):

[0165]

[0166] Referring to the synthesis of compound (I-1), the yield was 43%.

[0167] 1H NMR (300 MHz, CDCl3) δ 9.15 (s, 1H), 9.11 (s, 1H), 8.78 - 8.76 (m, 1H), 8.54 (d, J = 3.5 Hz, 1H), 8.41 (d, J = 8.6 Hz, 1H), 8.35 - 8.26 (m, 3H), 7.73 (dd, J = 8.6, 2.3 Hz, 1H), 3.52 (s, 2H), 2.58 - 2.47 (m, 10H), 1.14 (t, J = 7.2 Hz, 3H).

[0168] Example 3:

[0169] Synthesis of tert-butyl 4-((6-((4-(benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)methyl)piperazine-1-carboxylate(I-3):

[0170]

[0171] Referring to the synthesis of compound (I-1), the yield was 52%.

[0172] 1 H NMR (400 MHz, CDCl3) δ 9.14 (s, 1H), 8.83 (s, 1H), 8.77 (d, J = 1.5 Hz, 1H), 8.52 (d, J = 3.5 Hz, 1H), 8.41 (d, J = 8.6 Hz, 1H), 8.34 - 8.27 (m, 3H), 7.75 - 7.73 (m, 1H), 3.50 (s, 2H), 3.45 - 3.43 (m, J = 5.2 Hz, 4H), 2.43 - 2.40 (m, 4H), 1.46 (s, 9H).

[0173] Example 4:

[0174] Synthesis of 4-(benzothiazole-6-yl)-5-fluoro-N-(5-(piperazine-1-ylmethyl)pyridine-2-yl)pyrimidine-2-amine hydrochloride (I-4):

[0175]

[0176] tert-butyl 4-((6-((4-(benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)methyl)piperazine-1-carboxylate was dissolved in dichloromethane and reacted for 2 hours under 0°C conditions with HCl gas introduced. After the reaction was finished, the mixture was concentrated to obtain the compound 4-(benzothiazole-6-yl)-5-fluoro-N-(5-(piperazine-1-ylmethyl)pyridine-2-yl)pyrimidine-2-amine hydrochloride, with a yield of 100%.

[0177] 1 ¹H NMR (300 MHz, DMSO- d 6) δ 12.17 (s, 1H), 10.08 (s, 1H), 9.62 (s, 1H), 8.95 (t, J = 2.7 Hz, 2H), 8.71 (s, 1H), 8.53 (d, J = 8.8 Hz, 1H), 8.30 (q, J = 8.7 Hz, 2H), 7.99 (d, J = 9.0 Hz, 1H), 4.57 (s, 2H), 3.54 - 3.42 (m, 10H).

[0178] Example 5:

[0179] Synthesis of tert-butyl 4-(6-((4-(benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)nicotinoyl)piperazine-1-carboxylate(I-5):

[0180]

[0181] Referring to the synthesis of compound (I-1), the yield was 52%.

[0182] 1 H NMR (400 MHz, CDCl3) δ 9.16 (s, 1H), 8.78 - 8.78 (m, 1H), 8.52 - 8.49 (m, 2H), 8.44 (dd, J= 2.4, 0.8 Hz, 1H), 8.34 - 8.28 (m, 3H), 7.84 (dd, J = 8.7, 2.3 Hz, 1H), 3.70 - 3.44 (m, 8H), 1.48 (s, 9H).

[0183] Example 6:

[0184] Synthesis of 6-((4-(benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)(piperazine-1-yl)ketone hydrochloride (I-6):

[0185]

[0186] Referring to the synthesis of compound (I-4), the yield of 6-((4-(benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)(piperazine-1-yl)ketone hydrochloride was 100%.

[0187] 1 ¹H NMR (400 MHz, DMSO- d 6) δ 11.25 (s, 1H), 9.60 - 9.58 (m, 3H), 8.93 (d, J = 1.6 Hz, 1H), 8.88 (d, J = 3.3 Hz, 1H), 8.51 (d, J = 2.2 Hz, 1H), 8.32 - 8.25 (m, 2H), 8.19 - 8.09 (m, 2H), 3.79 - 3.76 (m, 4H), 3.18 - 3.16 (m, 4H).

[0188] Example 7:

[0189] Synthesis of tert-butyl 4-(6-((4-(benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)piperazine-1-carboxylate(I-7):

[0190]

[0191] Referring to the synthesis of compound (I-1), the yield was 47%.

[0192] 1H NMR (400 MHz, CDCl3) δ 9.15 (s, 1H), 8.77 - 8.76 (m, 1H), 8.45 (d, J = 3.4 Hz, 1H), 8.33 - 8.26 (m, 3H), 8.04 - 8.02 (m, 2H), 7.38 (dd, J = 9.1, 3.0 Hz, 1H), 3.63 - 3.60 (m, 4H), 3.11 - 3.09 (m, 4H), 1.49 (s, 9H).

[0193] Example 8:

[0194] Synthesis of 4-(benzothiazole-6-yl)-5-fluoro-N-(5-(piperazine-1-yl)pyridine-2-yl)pyrimidine-2-amine hydrochloride (I-8):

[0195]

[0196] Referring to the synthesis of compound (I-4), the yield was 100%.

[0197] 1 ¹H NMR (400 MHz, DMSO- d 6) δ 11.43 (s, 1H), 9.60 (s, 1H), 9.33 (s, 2H), 8.91 (s, 1H), 8.87 (d, J = 3.3 Hz, 1H), 8.33 - 8.23 ​​(m, 2H), 8.07 (d, J = 9.4 Hz, 1H), 8.01 (d, J = 2.9 Hz, 1H), 7.85 (d, J = 9.4 Hz, 1H), 3.44 (t, J = 5.1 Hz, 4H), 3.27 - 3.25 (m, 4H).

[0198] Example 9:

[0199] Synthesis of 4-(benzothiazole-6-yl)-5-fluoro-N-(5-((4-(methanesulfonyl)piperazine-1-yl)methyl)pyridine-2-yl)pyrimidine-2-amine(I-9):

[0200]

[0201] Referring to the synthesis of compound (I-1), the yield was 45%.

[0202] 1 H NMR (400 MHz, CDCl3) δ 9.15 (s, 1H), 8.78 - 8.77 (m, 1H), 8.48 (d, J = 3.4 Hz, 1H), 8.40 (dd, J = 8.5, 0.8 Hz, 1H), 8.34 - 8.29 (m, 2H), 8.25 - 8.25 (m, 1H), 8.18 (s, 1H), 7.70 (dd, J = 8.6, 2.4 Hz, 1H), 3.53 (s, 2H), 3.25 (t, J = 4.9 Hz, 4H), 2.78 (s, 3H), 2.58 (t, J = 5.0 Hz, 4H).

[0203] Example 10:

[0204] Synthesis of ethyl 4-((6-((4-(benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)methyl)piperazine-1-carboxylate(I-10):

[0205]

[0206] Referring to the synthesis of compound (I-1), the yield was 55%.

[0207] 1 H NMR (400 MHz, CDCl3) δ 9.15 (s, 1H), 8.78 - 8.77 (m, 1H), 8.48 (d, J = 3.4 Hz, 1H), 8.39 (dd, J = 8.6, 0.8 Hz, 1H), 8.34 - 8.27 (m, 2H), 8.25 - 8.24 (m, 2H), 7.72 (dd, J = 8.6, 2.3 Hz, 1H), 4.13 (q, J = 7.1 Hz, 2H), 3.50 - 3.47 (m, 6H), 2.42 (t, J = 5.0 Hz, 4H), 1.26 (t, J) = 7.1 Hz, 3H).

[0208] Example 11:

[0209] Synthesis of 4-(benzothiazole-6-yl)-5-fluoro-N-(5-((4-isopropylpiperazine-1-yl)methyl)pyridine-2-yl)pyrimidine-2-amine(I-11):

[0210]

[0211] Referring to the synthesis of compound (I-1), the yield was 48%.

[0212] 1 H NMR (400 MHz, CDCl3) δ 9.15 (s, 1H), 8.78 (d, J = 1.6 Hz, 1H), 8.48 (d, J = 3.5 Hz, 1H), 8.38 (dd, J = 8.5, 0.8 Hz, 1H), 8.35 - 8.32 (m, 1H), 8.30 - 8.27 (m, 1H), 8.25 - 8.23 ​​(m, 2H), 7.73 (dd, J = 8.6, 2.3 Hz, 1H), 3.50 (s, 2H), 2.68 - 2.44 (m, 9H), 1.06 (d, J = 6.5 Hz, 6H).

[0213] Example 12:

[0214] Synthesis of (4-ethylpiperazine-1-yl)(6-((5-fluoro-4-(2-methylbenzothiazole-6-yl)pyrimidine-2-yl)amino)pyridine-3-yl)ketone(I-12):

[0215]

[0216] Referring to the synthesis of compound (I-1), the yield was 40%.

[0217] 1H NMR (400MHz, CDCl3) δ 9.25(s, 1H), 8.63(d, J = 1.8Hz, 1H), 8.56 - 8.54(m, 2H), 8.51(d, J = 8.7Hz, 1H), 8.24(dt, J = 8.7, 1.3Hz, 1H), 8.09(d, J= 8.6Hz, 1H), 7.86(dd, J = 8.7, 2.4Hz, 1H), 3.78 - 3.64(m, 4H), 2.90 (s, 3H), 2.51 - 2.46(m, 6H), 1.13(t, J = 7.1Hz, 3H).

[0218] Example 13:

[0219] Synthesis of N-(5-((4-ethylpiperazine-1-yl)methyl)pyridine-2-yl)-5-fluoro-4-(2-methylbenzothiazole-6-yl)pyrimidine-2-amine(I-13):

[0220]

[0221] Referring to the synthesis of compound (I-1), the yield was 52%.

[0222] 1 H NMR (300 MHz, CDCl3) δ 8.63 (d, J = 1.8 Hz, 1H), 8.49 - 8.47 (m, 2H), 8.38 (d, J = 8.6 Hz, 1H), 8.28 - 8.24 (m, 2H), 8.08 (d, J = 8.7 Hz, 1H), 7.72 (dd, J = 8.6, 2.3 Hz, 1H), 3.51 (s, 2H), 2.90 (s, 3H), 2.55 - 2.45 (m, 10H), 1.11 (t, J = 7.2 Hz, 3H).

[0223] Example 14:

[0224] Synthesis of tert-butyl 4-((6-((5-fluoro-4-(2-methylbenzothiazole-6-yl)pyrimidine-2-yl)amino)pyridine-3-yl)methyl)piperazine-1-carboxylate(I-14):

[0225]

[0226] Referring to the synthesis of compound (I-1), the yield was 55%.

[0227] 1H NMR (300 MHz, CDCl3) δ 8.63 (s, 1H), 8.46 (d, J = 3.5 Hz, 1H), 8.39 (d, J = 8.6 Hz, 1H), 8.32 (s, 1H), 8.27 - 8.24 (m, 2H), 8.08 (d, J = 8.7 Hz, 1H), 7.72 (d, J = 8.4 Hz, 1H), 3.49 (s, 2H), 3.43 (t, J = 4.8 Hz, 4H), 2.91 (s, 3H), 2.42 - 2.39 (m, 4H), 1.46 (s, 9H).

[0228] Example 15:

[0229] Synthesis of 5-fluoro-4-(2-methylbenzothiazole-6-yl)-N-(5-(piperazine-1-ylmethyl)pyridine-2-yl)pyrimidine-2-amine hydrochloride (I-15):

[0230]

[0231] Referring to the synthesis of compound (I-4), the yield was 100%.

[0232] 1 ¹H NMR (400 MHz, DMSO- d 6) δ 11.67 (s, 1H), 9.90 (s, 2H), 8.89 (d, J = 3.3 Hz, 1H), 8.80 (d, J = 1.7 Hz, 1H), 8.64 (d, J = 2.2 Hz, 1H), 8.40 (dd, J = 9.0, 2.2 Hz, 1H), 8.21 (dt, J = 8.6, 1.3 Hz, 1H), 8.13 (d, J = 8.6 Hz, 1H), 8.04 (d, J = 8.9 Hz, 1H), 4.50 (s, 2H), 3.50 - 3.45 (m, 8H), 2.88 (s, 3H).

[0233] Example 16:

[0234] Synthesis of tert-butyl 4-(6-((5-fluoro-4-(2-methylbenzothiazole-6-yl)pyrimidine-2-yl)amino)nicotinoyl)piperazine-1-carboxylate(I-16):

[0235]

[0236] Referring to the synthesis of compound (I-1), the yield was 40%.

[0237] 1 H NMR (400 MHz, CDCl3) δ 9.11 (s, 1H), 8.63 (s, 1H), 8.55 - 8.51 (m, 3H), 8.24 (d, J = 8.6 Hz, 1H), 8.09 (dd, J = 8.7, 3.3 Hz, 1H), 7.86 (dt, J = 8.9, 2.6 Hz, 1H), 3.67 - 3.46 (m, 8H), 2.91 (s, 3H), 1.48 (s, 9H).

[0238] Example 17:

[0239] Synthesis of (6-((5-fluoro-4-(2-methylbenzothiazole-6-yl)pyrimidine-2-yl)amino)pyridine-3-yl)(piperazine-1-yl)ketone hydrochloride (I-17):

[0240]

[0241] Referring to the synthesis of compound (I-4), the yield was 100%.

[0242] 1 ¹H NMR (300 MHz, DMSO- d 6) δ 12.04 (s, 1H), 9.85 (s, 2H), 8.92 (d, J = 2.9 Hz, 1H), 8.81 (s, 1H), 8.60 (s, 1H), 8.28 (d, J = 9.1 Hz, 1H), 8.21 (d, J = 8.7 Hz, 1H), 8.13 (d, J = 8.6 Hz, 1H), 8.06 (d,J = 9.0 Hz, 1H), 3.83 - 3.77 (m, 4H), 3.20 - 3.14 (m, 4H), 2.88 (s, 3H).

[0243] Example 18:

[0244] Synthesis of (6-((4-(2-(cyclopentylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)(4-ethylpiperazine-1-yl)ketone(I-18):

[0245]

[0246] Referring to the synthesis of compound (I-1), the yield was 45%.

[0247] 1 H NMR (400 MHz, CDCl3) δ 8.90 (s, 1H), 8.50 (d, J = 8.7 Hz, 1H), 8.46 (d, J = 2.2 Hz, 1H), 8.43 (d, J = 3.6 Hz, 1H), 8.40 (d, J = 1.8 Hz, 1H), 8.07 (dd, J = 8.7, 1.8 Hz, 1H), 7.84 (dd, J = 8.7, 2.3 Hz, 1H), 7.64 (d, J = 8.5 Hz, 1H), 6.08 (s, 1H), 3.80 - 3.65 (m, 4H), 2.52 - 2.47 (m, 6H), 2.18 - 2.12 (m, 2H), 1.81 - 1.64 (m, 6H), 1.13 (t, J = 7.2 Hz, 3H).

[0248] Example 19:

[0249] Synthesis of N-cyclopentyl-6-(2-((5-((4-ethylpiperazine-1-yl)methyl)pyridine-2-yl)amino)-5-fluoropyrimidine-4-yl)benzothiazole-2-amine(I-19):

[0250]

[0251] Referring to the synthesis of compound (I-1), the yield was 51%.

[0252] 1H NMR (400 MHz, CDCl3) δ 8.59 (s, 1H), 8.43 - 8.35 (m, 3H), 8.23 ​​(d, J = 2.2 Hz, 1H), 8.08 - 8.05 (m, 1H), 7.71 (dd, J = 8.6, 2.3 Hz, 1H), 7.64 (d, J = 8.6 Hz, 1H), 6.12 (d, J = 6.7 Hz, 1H), 4.08 - 4.04 (m, 1H), 3.50 (s, 2H), 2.54 - 2.42 (m, 10H), 2.19 - 2.11 (m, 2H), 1.79 - 1.64 (m, 6H), 1.10 (t, J = 7.2 Hz, 3H).

[0253] Example 20:

[0254] Synthesis of tert-butyl 4-((6-((4-2-(cyclopentylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)methyl)piperazine-1-carboxylate(I-20):

[0255]

[0256] Referring to the synthesis of compound (I-1), the yield was 57%.

[0257] 1 H NMR (400 MHz, CDCl3) δ 8.43 - 8.40 (m, 2H), 8.38 (d, J = 3.7 Hz, 1H), 8.22 (s, 2H), 8.11 (d, J = 7.8 Hz, 1H), 7.79 (s, 1H), 7.63 (d, J = 8.4 Hz, 1H), 5.71 (s, 1H), 4.13 - 4.05 (m, 1H), 3.60 - 3.45 (m, 6H), 2.57 - 2.43 (m, 4H), 2.20 - 2.12 (m, 2H), 1.80 - 1.61 (m, 6H), 1.46 (s, 9H).

[0258] Example 21:

[0259] Synthesis of N-cyclopentyl-6-(5-fluoro-2-((5-(piperazine-1-ylmethyl)pyridine-2-yl)amino)pyrimidine-4-yl)benzothiazole-2-amine hydrochloride (I-21):

[0260]

[0261] Referring to the synthesis of compound (I-4), the yield was 100%.

[0262] 1 ¹H NMR (400 MHz, DMSO- d 6) δ 11.31 (s, 1H), 9.80 (s, 2H), 9.40 (s, 1H), 8.79 (d, J = 3.6 Hz, 1H), 8.60 (d, J = 2.2 Hz, 1H), 8.53 (d, J = 1.8 Hz, 1H), 8.32 (dd, J = 9.1, 2.2 Hz, 1H), 8.10 - 8.06 (m, 2H), 7.65 (d, J = 8.6 Hz, 1H), 4.44 (s, 2H), 4.29 - 4.24 (m, 1H), 3.46 - 3.38 (m, 8H), 3.17 (s, 1H), 2.06 - 1.99 (m, 2H), 1.73 - 1.59 (m, 6H).

[0263] Example 22:

[0264] Synthesis of tert-butyl 4-(6-((4-(2-(cyclopentylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)nicotinoyl)piperazine-1-carboxylate(I-22):

[0265]

[0266] Referring to the synthesis of compound (I-1), the yield was 53%.

[0267] 1 H NMR (400 MHz, CDCl3) δ 8.82 (s, 1H), 8.51 (d, J= 8.4 Hz, 1H), 8.46 - 8.41 (m, 3H), 8.09 (d, J = 8.3 Hz, 1H), 7.85 - 7.82 (m, 1H), 7.63 (d, J = 8.3 Hz, 1H), 6.20 (s, 1H), 4.09 - 4.03 (m, 1H), 3.68 - 3.45 (m, 8H), 2.18 - 2.12 (m, 2H), 1.80 - 1.62 (m, 6H), 1.48 (s, 9H).

[0268] Example 23:

[0269] Synthesis of (6-((4-(2-(cyclopentylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)(piperazine-1-yl)ketone hydrochloride (I-23):

[0270]

[0271] Referring to the synthesis of compound (I-4), the yield was 100%.

[0272] 1 ¹H NMR (400 MHz, DMSO- d 6) δ 11.15 (s, 1H), 9.70 (s, 1H), 9.52 (s, 2H), 8.80 (s, 1H), 8.56 - 8.50 (m, 2H), 8.15 - 8.09 (m, 3H), 7.69 (d, J = 8.6 Hz, 1H), 4.32 - 4.25 (m, 1H), 3.80 - 3.74 (m, 4H), 3.33 (s, 1H), 3.20 - 3.14 (m, 4H), 2.05 - 1.99 (m, 2H), 1.76 - 1.59 (m, 6H).

[0273] Example 24:

[0274] Synthesis of ethyl 4-((6-((4-(2-(cyclopentylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)methyl)piperazine-1-carboxylate(I-24):

[0275]

[0276] Referring to the synthesis of compound (I-1), the yield was 56%. 1 H NMR (400 MHz, CDCl3) δ 8.82 (s, 1H), 8.42 - 8.38 (m,3H), 8.25 - 8.24 (m, 1H), 8.05 (d, J = 8.3 Hz, 1H), 7.73 (d, J = 8.4 Hz, 1H), 7.64 (d, J = 8.3 Hz, 1H), 6.25 (s, 1H), 4.14 (q, J = 7.0 Hz, 2H), 4.08 - 4.02 (m,1H), 3.54 - 3.47 (m, 6H), 2.46 - 2.42 (m, 4H), 2.19 - 2.11 (m, 2H), 1.79 - 1.61 (m, 6H), 1.26 (t, J = 7.0 Hz, 3H).

[0277] Example 25:

[0278] Synthesis of N-cyclopentyl-6-(5-fluoro-2-((5-((4-(methanesulfonyl)piperazine-1-yl)methyl)pyridine-2-yl)amino)pyrimidine-4-yl)benzothiazole-2-amine(I-25):

[0279]

[0280] Referring to the synthesis of compound (I-1), the yield was 53%.

[0281] 1H NMR (400 MHz, CDCl3) δ 8.54 (s, 1H), 8.42 - 8.39 (m, 3H), 8.24 (d, J = 2.3 Hz, 1H), 8.10 - 8.07 (m, 1H), 7.69 (dd, J = 8.6, 2.3 Hz, 1H), 7.64 (d, J = 8.6 Hz, 1H), 5.96 (d, J = 6.8 Hz, 1H), 4.09 - 4.04 (m, 1H), 3.52 (s, 2H), 3.25 (t, J = 4.9 Hz, 4H), 2.79 (s, 3H), 2.58 (t, J = 4.9 Hz, 4H), 2.19 - 2.11 (m, 2H), 1.80 - 1.70 (m, 6H).

[0282] Example 26:

[0283] Synthesis of tert-butyl 4-(6-((4-(2-(cyclopentylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)piperazine-1-carboxylate(I-26):

[0284]

[0285] Referring to the synthesis of compound (I-1), the yield was 44%.

[0286] 1 H NMR (400 MHz, CDCl3) δ8.48 (s, 1H), 8.39 (d, J = 1.8 Hz, 1H), 8.36 (d, J = 3.7 Hz, 1H), 8.31 (d, J = 9.1 Hz, 1H), 8.08 - 8.05 (m, 1H), 8.03 (d, J = 2.9 Hz, 1H), 7.63 (d, J = 8.5 Hz, 1H), 7.37 (dd, J = 9.1, 3.0 Hz, 1H), 6.10 (d, J = 6.6 Hz, 1H), 4.08 - 4.03 (m, 1H), 3.61 (t, J = 5.1 Hz, 4H), 3.09 (t, J = 5.0 Hz, 4H), 2.17 - 2.11 (m, 2H), 1.77 - 1.64 (m, 6H), 1.49 (s, 9H).

[0287] Example 27:

[0288] Synthesis of N-cyclopentyl-6-(5-fluoro-2-((5-(piperazine-1-yl)pyridine-2-yl)amino)pyrimidine-4-yl)benzothiazole-2-amine hydrochloride (I-27):

[0289]

[0290] Referring to the synthesis of compound (I-4), the yield was 100%.

[0291] 1 ¹H NMR (400 MHz, DMSO- d 6) δ 11.97 (s, 1H), 10.53 (s, 1H), 9.86 (s, 2H), 8.83 (d, J = 3.3 Hz, 1H), 8.57 (d, J = 1.7 Hz, 1H), 8.27 (dd, J = 9.6, 2.6 Hz, 1H), 8.11 - 8.09 (m, 1H), 8.05 (d, J = 2.6 Hz, 1H), 7.86 (d, J = 9.5 Hz, 1H), 7.77 (d, J = 8.6 Hz, 1H), 4.41 - 4.35 (m, 1H), 3.52 - 3.50 (m, 4H), 3.26 - 3.23 (m, 4H), 2.08 - 2.02 (m, 2H), 1.76 - 1.60 (m, 6H).

[0292] Example 28:

[0293] Synthesis of N-cyclopentyl-6-(5-fluoro-2-((5-((4-isopropylpiperazine-1-yl)methyl)pyridine-2-yl)amino)pyrimidine-4-yl)benzothiazole-2-amine(I-28):

[0294]

[0295] Referring to the synthesis of compound (I-1), the yield was 50%.

[0296] 1H NMR (400 MHz, CDCl3) δ8.78 (s, 1H), 8.40 - 8.38 (m, 3H), 8.23 ​​(d, J = 2.2 Hz, 1H), 8.05 (dd, J = 8.5, 1.8 Hz, 1H), 7.71 (dd, J = 8.6, 2.3 Hz, 1H), 7.65 (d, J = 8.6 Hz, 1H), 6.32 (d, J = 6.2 Hz, 1H), 4.07 - 4.02 (m, 1H), 3.49 (s, 2H), 2.70 - 2.55 (m, 9H), 2.18 - 2.11 (m, 2H), 1.80 - 1.64 (m, 6H), 1.06 (d, J = 6.5 Hz, 6H).

[0297] Example 29:

[0298] Synthesis of (6-((4-(2-(cyclopentylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)(4-isopropylpiperazine-1-yl)ketone(I-29):

[0299]

[0300] Referring to the synthesis of compound (I-1), the yield was 48%.

[0301] 1 H NMR (400 MHz, CDCl3) δ8.78 (s, 1H), 8.40 - 8.38 (m, 3H), 8.23 ​​(d, J = 2.2 Hz, 1H), 8.05 (dd, J = 8.5, 1.8 Hz, 1H), 7.71 (dd, J = 8.6, 2.3 Hz, 1H), 7.65 (d, J = 8.6 Hz, 1H), 6.32 (d, J = 6.2 Hz, 1H), 4.07 - 4.02 (m, 1H), 3.49 (s, 2H), 2.70 - 2.55 (m, 9H), 2.18 - 2.11 (m, 2H), 1.80 - 1.64 (m, 6H), 1.06 (d, J = 6.5 Hz, 6H).

[0302] Example 30:

[0303] Synthesis of 6-(2-((5-((4-ethylpiperazine-1-yl)methyl)pyridine-2-yl)amino)-5-fluoropyrimidine-4-yl)-N,N-dimethylbenzothiazole-2- Synthesis of amine (I-30):

[0304]

[0305] Referring to the synthesis of compound (I-1), the yield was 42%.

[0306] 1 H NMR (400 MHz, CDCl3) δ 8.45 (d, J = 1.8 Hz, 1H), 8.38 - 8.36 (m, 2H), 8.21 (d, J = 2.3 Hz, 1H), 8.15 (dt, J = 8.7, 1.2 Hz, 1H), 7.94 (s, 1H), 7.71 (dd, J = 8.6, 2.3 Hz, 1H), 7.65 (d, J = 8.6 Hz, 1H), 3.49 (s, 2H), 3.27 (s, 6H), 2.60 - 2.36 (m, 10H), 1.09 (t, J = 7.2 Hz, 3H).

[0307] Example 31:

[0308] Synthesis of (6-((4-(2-(dimethylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)(4-ethylpiperazine-1-yl)ketone(I-31):

[0309]

[0310] 6-(2-chloro-5-fluoropyrimidine-4-yl)-N,N-dimethylbenzothiazole-2-amine (154 mg, 0.5 mmol) and 6-aminopyridine-3-yl)(4-ethylpiperazine-1-yl) ketone (141 mg, 0.6 mmol) were dissolved in dioxane (5 mL). Subsequently, Pd2(dba)3 (23 mg, 0.025 mmol), BINAP (31 mg, 0.05 mmol), and sodium tert-butoxide (96 mg, 1.0 mmol) were added. The reaction was carried out for 12 hours at 100°C, with the argon being switched three times. The mixture was cooled, filtered, and concentrated, and column chromatography (DCM~DCM / MeOH=10:1) was performed to obtain the compound (6-((4-(2-(dimethylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)(4-ethylpiperazine-1-yl)ketone (139 mg, 55% yield).

[0311] 1 ¹H NMR (400 MHz, DMSO- d 6) δ 10.27 (s, 1H), 8.69 (d, J = 3.8 Hz, 1H), 8.52 (d, J = 2.0 Hz, 1H), 8.35 (d, J = 2.4 Hz, 1H), 8.31 (d, J = 8.7 Hz, 1H), 8.06 (d, J = 8.6 Hz, 1H), 7.86 (dd, J = 8.6, 2.4 Hz, 1H), 7.59 (d, J = 8.6 Hz, 1H), 3.59 - 3.47 (m, 4H), 3.21 (s, 6H), 2.44 - 2.34 (m, 6H), 1.01 (t, J = 7.1 Hz, 3H).

[0312] Example 32:

[0313] Synthesis of tert-butyl 4-((6-((4-(2-(dimethylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)methyl)piperazine-1-carboxylate(I-32):

[0314]

[0315] Referring to the synthesis of compound (I-1), the yield was 48%.

[0316] 1 H NMR (400 MHz, CDCl3) δ 8.45 (d, J = 1.8 Hz, 1H), 8.40 - 8.36 (m, 2H), 8.20 (d, J = 2.2 Hz, 1H), 8.15 (dt, J = 8.8, 1.2 Hz, 1H), 7.94 (s, 1H), 7.71 (dd, J = 8.6, 2.3 Hz, 1H), 7.65 (d, J = 8.6 Hz, 1H), 3.48 (s, 2H), 3.45 - 3.42 (m, 4H), 3.27 (s, 6H), 2.41 - 2.39 (m, 4H), 1.46 (s, 9H).

[0317] Example 33:

[0318] Synthesis of 6-(5-fluoro-2-((5-(piperazine-1-ylmethyl)pyridine-2-yl)amino)pyrimidine-4-yl)-N,N-dimethylbenzothiazole-2-amine hydrochloride (I-33):

[0319]

[0320] Referring to the synthesis of compound (I-4), the yield was 100%.

[0321] 1 ¹H NMR (300 MHz, DMSO- d 6) δ 11.44 (s, 1H), 9.83 (s, 2H), 8.79 (d, J = 2.9 Hz, 1H), 8.59 (d, J = 11.9 Hz, 2H), 8.35 (d, J= 8.9 Hz, 1H), 8.07 (dd, J = 14.1, 8.8 Hz, 2H), 7.65 (d, J = 8.6 Hz, 1H), 4.46 (s, 2H), 3.49 - 3.41 (m, 8H), 3.24 (s, 6H).

[0322] Example 34:

[0323] Synthesis of tert-butyl 4-(6-((4-(2-(dimethylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)nicotinoyl)piperazine-1-carboxylate(I-34):

[0324]

[0325] Referring to the synthesis of compound (I-1), the yield was 50%.

[0326] 1 H NMR (400 MHz, CDCl3) δ 8.91 (s, 1H), 8.53 (dd, J = 8.6, 0.8 Hz, 1H), 8.50 (dd, J = 2.4, 0.8 Hz, 1H), 8.45 (d, J = 3.8 Hz, 1H), 8.44 (d, J = 1.8 Hz, 1H), 8.16 - 8.13 (m, 1H), 7.84 (dd, J = 8.7, 2.4 Hz, 1H), 7.66 (d, J = 8.6 Hz, 1H), 3.70 - 3.59 (m, 4H), 3.51 - 3.46 (m, 4H), 3.27 (s, 6H), 1.48 (s, 9H).

[0327] Example 35:

[0328] Synthesis of (6-((4-(2-(dimethylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)(piperazine-1-yl)ketone hydrochloride (I-35):

[0329]

[0330] Referring to the synthesis of compound (I-1), the yield was 100%.

[0331] 1 H NMR (300 MHz, CDCl3) δ 11.70 (s, 1H), 9.84 (s, 1H), 9.71 (s, 1H), 8.83 (s, 1H), 8.58 (d, J = 17.9 Hz, 2H), 8.21 (d, J = 8.9 Hz, 1H), 8.14 - 8.05 (m, 2H), 7.70 (d, J = 8.6 Hz, 1H), 3.35 - 3.32 (m, 4H), 3.28 (s, 6H), 3.20 - 3.15 (m, 4H).

[0332] Example 36:

[0333] Synthesis of 6-(5-fluoro-2-((5-((4-isopropylpiperazine-1-yl)methyl)pyridine-2-yl)amino)pyrimidine-4-yl)-N,N-dimethylbenzothiazole-2-amine(I-36):

[0334]

[0335] Referring to the synthesis of compound (I-1), the yield was 52%.

[0336] 1 H NMR (400 MHz, CDCl3) δ 8.45 (d, J = 1.8 Hz, 1H), 8.38 (dd, J = 6.3, 2.4 Hz, 2H), 8.23 ​​(d, J = 2.3 Hz, 1H), 8.16 - 8.14 (m, 2H), 7.71 (dd, J = 8.6, 2.3 Hz, 1H), 7.65 (d, J = 8.6 Hz, 1H), 3.49 (s, 2H), 3.27 (s, 6H), 2.68 - 2.54 (m, 9H), 1.05 (d, J = 6.5 Hz, 6H).

[0337] Example 37:

[0338] Synthesis of 6-(5-fluoro-2-((5-((4-(methanesulfonyl)piperazine-1-yl)methyl)pyridine-2-yl)amino)pyrimidine-4-yl)-N,N-dimethylbenzothiazole-2-amine(I-37):

[0339]

[0340] Referring to the synthesis of compound (I-1), the yield was 43%.

[0341] 1 H NMR (400 MHz, CDCl3) δ 8.44 - 8.35 (m, 3H), 8.21 (d, J = 7.7 Hz, 1H), 8.16 - 8.07 (m, 2H), 7.71 (s, 1H), 7.66 - 7.61 (m, 1H), 3.55 (s, 2H), 3.29 - 3.25 (m, 10H), 2.79 (s, 3H), 2.63 - 2.57 (m, 4H).

[0342] Example 38:

[0343] Synthesis of ethyl (6-((4-(2-(dimethylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)methyl)piperazine-1-carboxylate(I-38):

[0344]

[0345] Referring to the synthesis of compound (I-1), the yield was 53%.

[0346] 1 H NMR (400 MHz, CDCl3) δ 8.44 - 8.36 (m, 3H), 8.31 - 8.23 ​​(m, 2H), 8.15 (d, J = 8.3 Hz, 1H), 7.73 (s, 1H), 7.65 (d, J = 8.3 Hz, 1H), 4.13 (q, J = 7.1 Hz, 2H), 3.52 - 3.49 (m, 6H), 3.27 (s, 6H), 2.46 - 2.41 (m, 4H), 1.26 (t, J = 7.3 Hz, 3H).

[0347] Example 39:

[0348] Synthesis of (6-((4-(2-(dimethylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)(4-isopropylpiperazine-1-yl)ketone(I-39):

[0349]

[0350] Referring to the synthesis of compound (I-1), the yield was 44%.

[0351] 1 H NMR (400 MHz, CDCl3) δ 8.73 (s, 1H), 8.52 - 8.48 (m, 2H), 8.45 - 8.44 (m, 2H), 8.16 - 8.13 (m, 1H), 7.85 (dd, J = 8.7, 2.4 Hz, 1H), 7.65 (d, J = 8.6 Hz, 1H), 3.75 - 3.63 (m, 4H), 3.27 (s, 6H), 2.79 - 2.75 (m, 1H), 2.62 - 2.55 (m, 4H), 1.08 (d, J = 6.5 Hz, 6H).

[0352] Example 40:

[0353] Synthesis of tert-butyl 4-(6-((4-(2-(dimethylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)piperazine-1-carboxylate(I-40):

[0354]

[0355] Referring to the synthesis of compound (I-1), the yield was 40%.

[0356] 1 H NMR (400 MHz, CDCl3) δ 8.44 (s, 1H), 8.36 - 8.33 (m, 2H), 8.25 (s, 1H), 8.14 (dd, J = 8.7, 1.6 Hz, 1H), 8.00 (d, J = 2.8 Hz, 1H), 7.64 (dd,J = 8.6, 1.2 Hz, 1H), 7.40 (dd, J = 9.3, 2.9 Hz, 1H), 3.61 (t, J = 5.0 Hz, 4H), 3.27 (s, 3H), 3.09 (t, J = 5.0 Hz, 4H).

[0357] Example 41:

[0358] Synthesis of 6-(5-fluoro-2-((5-(piperazine-1-yl)pyridine-2-yl)amino)pyrimidine-4-yl)-N,N-dimethylbenzothiazole-2-amine hydrochloride (I-41):

[0359]

[0360] Referring to the synthesis of compound (I-4), the yield was 100%.

[0361] 1 ¹H NMR (400 MHz, DMSO- d 6) δ 11.99 (s, 1H), 9.77 (s, 2H), 8.81 (d, J = 3.5 Hz, 1H), 8.57 (d, J = 1.8 Hz, 1H), 8.28 (dd, J = 9.7, 2.8 Hz, 1H), 8.12 - 8.05 (m, 1H), 8.02 (d, J = 2.9 Hz, 1H), 7.82 (d, J = 9.6 Hz, 1H), 7.69 (d, J = 8.6 Hz, 1H), 3.49 (t, J = 5.1 Hz, 4H), 3.27 (s, 6H), 3.25 - 3.23 (m, 4H).

[0362] Example 42:

[0363] Synthesis of (6-((4-(2-(diethylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)(4-ethylpiperazine-1-yl)ketone(I-42):

[0364]

[0365] Referring to the synthesis of compound (I-1), the yield was 44%.

[0366] 1 H NMR (300 MHz, CDCl3) δ 8.56 (s, 1H), 8.50 (d, J = 8.7 Hz, 1H), 8.47 (d, J = 2.3 Hz, 1H), 8.43 - 8.41 (m, 2H), 8.13 (dd, J = 8.7, 1.7 Hz, 1H), 7.84 (dd, J = 8.7, 2.4 Hz, 1H), 7.62 (d, J = 8.6 Hz, 1H), 3.80 - 3.60 (m, 8H), 2.54 - 2.47 (m, 6H), 1.33 (t, J = 7.1 Hz, 6H), 1.13 (t, J = 7.1 Hz, 3H).

[0367] Example 43:

[0368] Synthesis of N,N-diethyl-6-(2-((5-((4-ethylpiperazine-1-yl)methyl)pyridine-2-yl)amino)-5-fluoropyrimidine-4-ylbenzothiazole-2-amine(I-43):

[0369]

[0370] Referring to the synthesis of compound (I-1), the yield was 38%.

[0371] 1 H NMR (400 MHz, CDCl3) δ 8.69 (s, 1H), 8.43 - 8.39 (m, 3H), 8.29 (d, J = 2.3 Hz, 1H), 8.15 - 8.12 (m, 1H), 7.71 (dd, J = 8.6, 2.3 Hz, 1H), 7.62 (d, J = 8.6 Hz, 1H), 3.63 (q, J= 7.2 Hz, 4H), 3.50 (s, 2H), 2.53 - 2.42 (m, 10H), 1.33 (t, J = 7.1 Hz, 6H), 1.10 (t, J = 7.2 Hz, 3H).

[0372] Example 44:

[0373] Synthesis of tert-butyl 4-((6-((4-(2-(diethylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)methyl)piperazine-1-carboxylate(I-44):

[0374]

[0375] Referring to the synthesis of compound (I-1), the yield was 48%.

[0376] 1 H NMR (400 MHz, CDCl3) δ 8.45 (s, 1H), 8.43 - 8.39 (m, 3H), 8.26 (d, J = 2.2 Hz, 1H), 8.15 - 8.12 (m, 1H), 7.73 (d, J = 8.2 Hz, 1H), 7.62 (d, J = 8.6 Hz, 1H), 3.63 (q, J = 7.1 Hz, 4H), 3.51 (s, 2H), 3.46 - 3.44 (m, 4H), 2.44 - 2.41 (m, 4H), 1.46 (s, 9H), 1.33 (t, J = 7.1 Hz, 6H).

[0377] Example 45:

[0378] Synthesis of N,N-diethyl-6-(5-fluoro-2-((5-(piperazine-1-ylmethyl)pyridine-2-yl)amino)pyrimidine-4-yl)benzothiazole-2-amine hydrochloride (I-45):

[0379]

[0380] Referring to the synthesis of compound (I-4), the yield was 100%.

[0381] 1 H NMR (400 MHz, CDCl3) δ 8.45 (s, 1H), 8.43 - 8.39 (m, 3H), 8.26 (d, J = 2.2 Hz, 1H), 8.15 - 8.12 (m, 1H), 7.73 (d, J = 8.2 Hz, 1H), 7.62 (d, J = 8.6 Hz, 1H), 3.63 (q, J = 7.1 Hz, 4H), 3.51 (s, 2H), 3.46 - 3.44 (m, 4H), 2.44 - 2.41 (m, 4H), 1.46 (s, 9H), 1.33 (t, J = 7.1 Hz, 6H).

[0382] Example 46:

[0383] Synthesis of tert-butyl 4-((6-((4-(2-(diethylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)nicotinoyl)piperazine-1-carboxylate(I-46):

[0384]

[0385] Referring to the synthesis of compound (I-1), the yield was 40%.

[0386] 1 H NMR (400 MHz, CDCl3) δ 8.99 (s, 1H), 8.53 (dd, J = 8.7, 0.8 Hz, 1H), 8.51 (dd, J = 2.4, 0.9 Hz, 1H), 8.45 (d, J = 3.8 Hz, 1H), 8.42 (d, J = 1.9 Hz, 1H), 8.15 - 8.12 (m, 1H), 7.84 (dd, J = 8.8, 2.4 Hz, 1H), 7.64 (d, J = 8.6 Hz, 1H), 3.67 - 3.61 (m, 8H), 3.50 - 3.47 (m, 4H), 1.48 (s, 9H), 1.34 (t, J= 7.1 Hz, 6H).

[0387] Example 47:

[0388] Synthesis of (6-((4-(2-(diethylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)(piperazine-1-yl)ketone(I-47):

[0389]

[0390] Referring to the synthesis of compound (I-4), the yield was 100%.

[0391] 1 ¹H NMR (400 MHz, DMSO- d 6) δ 11.78 (s, 1H), 10.03 (s, 1H), 9.88 (s, 2H), 8.83 (d, J = 3.6 Hz, 1H), 8.60 (d, J = 1.8 Hz, 1H), 8.58 (d, J = 2.1 Hz, 1H), 8.23 ​​(dd, J = 8.9, 2.2 Hz, 1H), 8.13 - 8.07 (m, 2H), 7.71 (d, J = 8.6 Hz, 1H), 3.86 - 3.78 (m, 3H), 3.67 (q, J = 7.1 Hz, 4H), 3.19 - 3.15 (m, 3H), 1.27 (t, J = 7.1 Hz, 6H).

[0392] Example 48:

[0393] Synthesis of N,N-diethyl-6-(5-fluoro-2-((5-((4-(methanesulfonyl)piperazine-1-yl)methyl)pyridine-2-yl)amino)pyrimidine-4-yl)benzothiazole-2-amine(I-48):

[0394]

[0395] Referring to the synthesis of compound (I-1), the yield was 44%.

[0396] 1H NMR (300 MHz, CDCl3) δ 8.92 (s, 1H), 8.44 - 8.42 (m, 3H), 8.32 - 8.21 (m, 1H), 8.13 (d, J = 8.6 Hz, 1H), 7.70 (dd, J = 8.6, 2.3 Hz, 1H), 7.62 (d, J = 8.7 Hz, 1H), 3.63 (q, J = 7.2 Hz, 4H), 3.52 (s, 2H), 3.25 (t, J = 4.8 Hz, 3H), 2.78 (s, 3H), 2.58 (t, J = 4.7 Hz, 4H), 1.33 (t, J = 7.1 Hz, 6H).

[0397] Example 49:

[0398] Synthesis of ethyl 4-((6-((4-(2-(diethylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)methyl)piperazine-1-carboxylate(I-49):

[0399]

[0400] Referring to the synthesis of compound (I-1), the yield was 50%.

[0401] 1 H NMR (300 MHz, CDCl3) δ 8.82 (s, 1H), 8.45 - 8.42 (m, 3H), 8.30 (d, J = 2.2 Hz, 1H), 8.13 (d, J = 8.6 Hz, 1H), 7.74 (d, J = 8.6 Hz, 1H), 7.62 (d, J = 8.6 Hz, 1H), 4.13 (q, J = 7.2 Hz, 2H), 3.63 (q, J = 7.2 Hz, 4H), 3.53 - 3.49 (m, 6H), 2.46 - 2.42 (m, 4H), 1.33 (t, J= 7.1 Hz, 6H), 1.26 (t, J = 7.1 Hz, 3H).

[0402] Example 50:

[0403] Synthesis of N,N-diethyl-6-5-fluoro-2-((5-((4-isopropylpiperazine-1-yl)methyl)pyridine-2-yl)amino)pyrimidine-4-yl)benzothiazole-2-amine(I-50):

[0404]

[0405] Referring to the synthesis of compound (I-1), the yield was 48%.

[0406] 1 H NMR (300 MHz, CDCl3) δ 9.05 (s, 1H), 8.45 - 8.40 (m, 3H), 8.32 (d, J = 2.3 Hz, 1H), 8.14 (d, J = 8.6 Hz, 1H), 7.71 (dd, J = 8.7, 2.3 Hz, 1H), 7.61 (d, J = 8.7 Hz, 1H), 3.62 (q, J = 7.2 Hz, 4H), 3.49 (s, 2H), 2.68 - 2.55 (m, 9H), 1.32 (t, J = 7.1 Hz, 6H), 1.05 (d, J = 6.4 Hz, 6H).

[0407] Example 51:

[0408] Synthesis of (6-((4-(2-(diethylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)(4-isopropylpiperazine-1-yl)ketone(I-51):

[0409]

[0410] Referring to the synthesis of compound (I-1), the yield was 47%.

[0411] 1H NMR (300 MHz, CDCl3) δ 9.69 (s, 1H), 8.60 (d, J = 2.3 Hz, 1H), 8.54 (d, J = 8.7 Hz, 1H), 8.50 (d, J = 3.7 Hz, 1H), 8.41 (s, 1H), 8.12 (d, J = 8.6 Hz, 1H), 7.86 (dd, J = 8.7, 2.3 Hz, 1H), 7.62 (d, J = 8.6 Hz, 1H), 3.77 - 3.59 (m, 8H), 2.78 - 2.72 (m, 1H), 2.59 - 2.54 (m, 4H), 1.32 (t, J = 7.1 Hz, 6H), 1.07 (d, J = 6.4 Hz, 6H).

[0412] Example 52:

[0413] Synthesis of tert-butyl 4-(6-((4-(2-(diethylamino)benzothiazole-6-yl)-5-fluoropyrimidine-2-yl)amino)pyridine-3-yl)piperazine-1-carboxylate(I-52):

[0414]

[0415] Referring to the synthesis of compound (I-1), the yield was 49%.

[0416] 1 H NMR (300 MHz, CDCl3) δ 8.58 (s, 1H), 8.41 (d, J = 1.8 Hz, 1H), 8.37 (d, J = 4.0 Hz, 1H), 8.34 (d, J = 9.1 Hz, 1H), 8.14 - 8.10 (m, 1H), 8.06 (d, J = 2.9 Hz, 1H), 7.61 (d, J = 8.6 Hz, 1H), 7.38 (dd, J= 9.1, 3.0 Hz, 1H), 3.66 - 3.58 (m, 8H), 3.09 (t, J = 5.1 Hz, 4H), 1.49 (s, 9H), 1.32 (t, J = 7.1 Hz, 6H).

[0417] Example 53:

[0418] Synthesis of N,N-diethyl-6-(5-fluoro-2-((5-(piperazine-1-yl)pyridine-2-yl)amino)pyrimidine-4-yl)benzothiazole-2-amine hydrochloride (I-53):

[0419]

[0420] Referring to the synthesis of compound (I-4), the yield was 100%.

[0421] 1 ¹H NMR (300 MHz, DMSO- d 6) δ 12.15 (s, 1H), 10.04 (s, 2H), 8.79 (s, 1H), 8.57 (s, 1H), 8.34 (d, J = 9.3 Hz, 1H), 8.08 - 8.05 (m, 2H), 7.87 (d, J = 8.9 Hz, 1H), 7.71 (d, J = 8.4 Hz, 1H), 3.69 - 3.67 (m, 4H), 3.56 - 3.53 (m, 4H), 3.27 - 3.23 (m, 4H), 1.30 - 1.25 (m, 6H).

[0422] In the above example, the corresponding pharmaceutically acceptable salt was reacted by dissolving the main product in dichloromethane and introducing HCl gas at 0°C for 2 hours. After the reaction is finished, the hydrochloride salt can be obtained by concentration.

[0423] II. Biological Analysis

[0424] (1) Method for analyzing and detecting CDK6 kinase activity

[0425] In this experiment, the Lance Ultra method from PerkinElmer Co., Ltd. was used for detection. Protein kinase, Ulight-labeled polypeptide substrate, ATP, and the compound were mixed on a test plate and the reaction was incubated. Subsequently, EDTA was added to stop the reaction, and a europium (Eu) chelate-labeled antibody was added for detection. The analysis of this experiment was performed using the Envision instrument from PerkinElmer Co., Ltd. in TR-FRET mode. After excitation at 320 / 340 nm, fluorescence signals were emitted at 665 nm and 615 nm wavelengths. Eu was transferred to an adjacent fluorophore ULight receptor via energy transfer, and the emitted light was detected.

[0426] Measured IC 50 The values ​​are shown in Table 1 below. From the experimental results, it can be seen that the compound of the embodiment of the present invention has strong inhibitory activity against CDK6 kinase activity.

[0427]

[0428] (2) DYRK2 kinase activity analysis and detection method

[0429] The DYRK2 kinase inhibitory activity of the compounds of the present invention was measured. The method was briefly described as follows (for specific methods, refer to the literature [Banerjee S, Wei T, Wang J, et al. Inhibition of dual-specificity tyrosine phosphorylation-regulated kinase 2 perturbs 26S proteasome-addicted neoplastic progression[J]. Proceedings of the National Academy of Sciences, 2019, 116(49): 24881-24891)]):

[0430] 1) Add compounds of different concentrations to a 384-well plate and redissolve, then DYRK2 protein, substrate Woodtide (KKISGRLSPIMTEQ), 33 P-γATP was added and mixed evenly.

[0431] 2) The mixture was incubated at room temperature for 30 minutes;

[0432] 3) After stopping the reaction by adding 0.5M (3%) orthophosphoric acid solution, the mixture was transferred to a P81 plate and washed with 50mM orthophosphoric acid solution.

[0433] 4) IC using GraphPad Prism software 50 Calculated the result.

[0434] IC 50 / The measured values ​​are shown in Table 2 below. From the experimental results, it can be seen that the compound of the embodiment of the present invention has strong inhibitory activity against DYRK2 kinase activity.

[0435]

[0436] (3) Check whether the proliferation of various cancer cells is inhibited

[0437] The inhibitory activity of compounds on the proliferation of 14 types of cells, including human breast cancer (MCF-7), triple-negative breast cancer (MDA-MB-231) cell line, multiple myeloma (RPMI8226) cell line, leukemia (K562) cell line, gastric cancer (MGC-803) cell line, ovarian cancer (SK-OV-3) cell line, colorectal cancer (HT-29) cell line, liver cancer (HepG2) cell line, pancreatic cancer (Panc-1) cell line, human glioma (U251) cell line, lung cancer (A-549), non-small cell lung cancer (NCI-H1299) cell line, and prostate cancer (PC-3, Du-145) cell line, was measured by the following method.

[0438] Experimental Protocol:

[0439] The inhibition of compounds on the proliferation of various cancer cells was measured according to the MTT method, and the half-inhibitory concentration (IC10) of the compounds for cell proliferation activity was determined. 50 I obtained.

[0440] 1) 1×10⁶ cells in the logarithmic phase in a 96-well plate 5 Cells were seeded per well and cultured at 37°C under 5% CO2 conditions until 90% fusion was achieved. Subsequently, the cells were synchronized by incubating for 2 hours in serum-free DMEM medium, RPMI-1640 medium, L-15 medium, F12K medium, MEM medium, F-12 medium, or IMDM medium (the corresponding medium was used for each cell).

[0441] 2) 100 μL of gradient dilution solutions of test compounds of different concentrations were added to a culture plate, and the culture plate was incubated in a 5% CO2 incubator at 37°C for 72 hours.

[0442] 3) 20 μL of MTT solution (5 mg / mL) was added to each well 4 hours before the end of culture. After incubation, the supernatant from each well was discarded, and 150 μL of DMSO was added to each well. The solution was vibrated on a cell oscillator for 10 minutes, and after the crystals were completely dissolved, the OD570 was measured using a microplate reader. Inhibition rate = (Control OD value - Experimental OD value) / Control OD value × 100%.

[0443] 4) After obtaining the data, fit it with GraphPad Prism 6 to create an IC 50 I obtained.

[0444] Various cancer cell proliferation activities were tested for the compound of Example 31 (I-31) and the commercially available CDK4 / 6 inhibitor palbociclib, and the measured IC50 50The values ​​are shown in Table 3. Compound I-31 exhibited inhibitory activity against the proliferation of 14 types of cells, including human breast cancer (MCF-7), triple-negative breast cancer (MDA-MB-231) cell line, multiple myeloma (RPMI8226) cell line, leukemia (K562) cell line, gastric cancer (MGC-803) cell line, ovarian cancer (SK-OV-3) cell line, colorectal cancer (HT-29) cell line, liver cancer (HepG2) cell line, pancreatic cancer (Panc-1) cell line, human glioma (U251) cell line, lung cancer (A-549), non-small cell lung cancer (NCI-H1299) cell line, and prostate cancer (PC-3, Du-145) cell line, and the inhibitory activity against the 14 types of cells was much stronger than that of palbociclib, a commercially available CDK4 / 6 inhibitor.

[0445]

[0446] (4) Determination of acute toxicity of the compound

[0447] Test animals: ICR mice; 18-22g; half males and half females; total 40 animals.

[0448] Dosage setting for each group: (1) Control group: One equal amount of normal saline was administered once by gastrointestinal feeding to 10 mice (half male, half female) in each group. (2) 2500 mg / kg group: One dose was administered once by gastrointestinal feeding to 10 mice (half male, half female). (3) 5000 mg / kg group: One dose was administered once by gastrointestinal feeding to 10 mice (half male, half female). (4) 10000 mg / kg group: One dose was administered once by gastrointestinal feeding to 10 mice (half male, half female).

[0449]

[0450] Laboratory environment: Room temperature 24±2℃, relative humidity 60–70%. Subjects of observation: The test drug (compound prepared in Example 31) was administered once according to the dosage in Table 4, and toxic symptoms and deaths in mice were recorded. The deceased animals were necropsied. The observation period was 14 days. As a result, no abnormalities were detected within 12 hours of administration in any group. No animals died within 24 hours of administration, nor did any animals die after 14 days of administration. No other obvious abnormalities were observed.

[0451] Changes in body weight are shown in Figure 1. No significant toxic effects were observed when 2500 mg / kg, 5000 mg / kg, or 10000 mg / kg were administered intragastically compared to the control group.

[0452] As shown in Figure 2 by the HE staining results, the compound (I-31) prepared in Example 31 did not show significant toxicity to the heart, liver, spleen, lungs, kidneys, and other major organs.

[0453] (5) Determination of pharmacokinetics of compounds

[0454] The weight of the test compound was measured and placed in a sterile vial, followed by the addition of 250 μL of DMSO and 10 μL of methanesulfonic acid. After dissolution, 4.78 mL of 5% glucose injection solution was added, and the mixture was uniformly mixed using ultrasound and a shaker to prepare a 2 mg / mL test compound solution, which was used as a gastric tube feeding agent. Additionally, 4.5 mL of 5% glucose injection solution was added to 0.5 mL of the 2 mg / mL test solution and mixed by shaking to prepare a 0.2 mg / mL test solution, which was used as an intravenous injection agent.

[0455] Six SD rats were divided into two groups. One was administered via tail vein (1 mg / kg), and the other was administered via gastrointestinal feeding with Example 31 (10 mg / kg). Approximately 0.25 mL of blood samples were collected from the retroorbital venous plexus at 2, 5, 15, 30, 1h, 2h, 4h, 6h, 8h, and 12h after administration for the intravenous group, and at 5, 15, 30, 1h, 2h, 4h, 6h, 8h, 12h, and 24h after administration for the gastrointestinal feeding group. The concentration of Example 31 in plasma samples from SD rats was determined by LC-MS / MS, pharmacokinetic parameters were calculated using WinNolin software, and the results are shown in Table 5.

[0456] The results show that the compound (I-31) of Example 31 of the present invention has good metabolism, good absorption and exposure, and high bioavailability in rats.

[0457]

[0458] (6) Measurement of the anti-lung cancer activity of the compound

[0459] The drug was compound (I-31) prepared in Example 31 with the commercially available CDK4 / 6 inhibitor palbociclib. Human non-small cell lung cancer cell line A-549 was cultured in RPMI-1640 medium containing 10% fetal bovine serum. The test animals were SPF grade BALB / c nude mice (5 males per group). The drug dosage settings are shown in Table 6.

[0460]

[0461] Drug formulation method:

[0462] Example 31 (150 mg / kg): 30 mg of the compound powder to be tested was weighed, dissolved in 2 mL of physiological saline, formulated into a 15 mg / mL drug, and administered orally by gastric tube feeding at a volume of 0.2 mL / 20 g.

[0463] Example 31 (300 mg / kg): 60 mg of the compound powder to be tested was weighed, dissolved in 2 ml of physiological saline, formulated into a 30 mg / mL drug, and administered orally at a volume of 0.2 mL / 20 g.

[0464] Palbociclib (150 mg / kg): 30 mg of the complex powder to be tested was weighed and dissolved in 2 mL of physiological saline, and prepared into a 15 mg / mL formulation for oral gastric tube administration at a dose of 0.2 mL / 20 g.

[0465] Experimental Methods: A nude mouse model of human lung cancer xenografts was established by inoculating the human lung cancer cell line A549 into the axillary skin of nude mice. Logarithmic-phase A549 cells were subcutaneously inoculated into the right axilla of 30 nude mice under sterile conditions, and the cell inoculation volume was 5 × 10⁶ 6 It was cell / mouse. The diameter of the xenograft was measured with a vernier caliper. The tumor was approximately 80 mm 3 When the mice had grown to a certain size, 20 tumor-bearing nude mice with good growth status and uniform tumor size were selected and randomly divided into four groups of four mice each: a model group, a low-dose group of Example 31 (150 mg / kg), a high-dose group of Example 31 (300 mg / kg), and a positive drug palbociclib (150 mg / kg). The test drugs Example 31 and palbociclib were administered intragastrically to the low-dose, high-dose, and positive drug groups once every two days. An equal amount of vehicle control was administered intragastrically to the model group. The antitumor effect of the test substances was observed dynamically by measuring tumor diameter. Tumor diameter was measured every other day, and the body weight of the nude mice was measured during the tumor diameter measurement. On day 22, the mice were sacrificed, and the surgically removed tumor fragments were fixed in 10% formaldehyde and stored in liquid nitrogen for later use.

[0466] Experimental results showed the relative tumor proliferation rate T / C (%) of the low-dose group (150 mg / kg) of Example 31 and the high-dose group (300 mg / kg) of Example 31 compared to the model group, which were 44.8% and 35.9%, respectively, and the tumor growth inhibition rates were 55.2% and 64.1%, respectively. When the positive drug palbociclib was administered via gastric tube feeding at a dose of 150 mg / kg, the relative tumor proliferation rate T / C (%) and tumor inhibition rate were 39.6% and 60.4%, respectively.

[0467] Therefore, the test drug prepared in Example 31 showed a significant inhibitory effect on the growth of human lung cancer A549 xenografts in nude mice, and the effect was superior to that of the positive control drug palbociclib.

[0468] (7) Determination of the prostate cancer (PC3) activity of the compound

[0469] The drug was compound (I-31) prepared in Example 31 with the commercially available CDK4 / 6 inhibitor palbociclib. The cell line was human prostate cancer PC-3 cells. The test animals were SPF-grade BALB / c nude mice; 8 males per group. The dosage settings are shown in Table 7.

[0470]

[0471] Drug manufacturing method:

[0472] Example 31 (100 mg / kg): 20 mg of the compound powder to be tested was weighed, dissolved in 2 mL of physiological saline, formulated into a 10 mg / mL drug, and administered orally by gastric tube feeding at a volume of 0.2 mL / 20 g.

[0473] Example 31 (200 mg / kg): 40 mg of the compound powder to be tested was weighed, dissolved in 2 ml of physiological saline, formulated into a 20 mg / mL drug, and administered orally by gastric tube feeding at a volume of 0.2 mL / 20 g.

[0474] Palbociclib (100 mg / kg): 20 mg of the compound powder to be tested was weighed and dissolved in 2 mL of physiological saline, and prepared as a 10 mg / mL formulation for oral gastric tube administration at a dose of 0.2 mL / 20 g.

[0475] Experimental Methods: A nude mouse model of human prostate cancer xenografts was established by inoculating human prostate cancer PC-3 cells subcutaneously into the axillary skin of nude mice. Logarithmic phase PC-3 cells were subcutaneously inoculated into the right axilla of 40 nude mice under sterile conditions, with a cell inoculation volume of 5 × 10⁶ cells. 6 It was cell / mouse. The diameter of the xenograft was measured with a vernier caliper. The tumor was approximately 90 mm 3 When the mice had grown to a certain size, 32 nude mice with tumors that were in good growth condition and had uniform tumor sizes were selected and randomly assigned 8 mice each to the model group, the low-dose group of Example 31 (100 mg / kg), the high-dose group of Example 31 (200 mg / kg), and the positive drug palbociclib (100 mg / kg) group. Test drugs Example 31 and palbociclib were administered intragastrically once daily to the low-dose, high-dose, and positive drug groups, respectively. An equal amount of vehicle control was administered intragastrically to the model group. The antitumor effect of the test substances was observed dynamically by measuring tumor diameter. Tumor diameters were measured every other day, and the body weight of the nude mice was measured during the tumor diameter measurement. On day 29, the mice were sacrificed, and the surgically removed tumor fragments were fixed in 10% formaldehyde and stored in liquid nitrogen for later use.

[0476] As a result of the experiment, the relative tumor proliferation rate T / C (%) of the low-dose group (100 mg / kg) of Example 31 and the high-dose group (200 mg / kg) of Example 31 compared to the model group were 35.7% and 23.4%, respectively, and the tumor growth inhibition rates were 64.3% and 76.6%, respectively. When palbociclib, a positive drug, was administered via gastric tube feeding at a dose of 100 mg / kg, the relative tumor proliferation rate T / C (%) and tumor inhibition rate were 35.5% and 64.5%, respectively.

[0477] Therefore, the test drug prepared in Example 31 showed a significant inhibitory effect on the growth of human prostate cancer PC3 xenografts in nude mice, and the effect was superior to that of the positive control drug palbociclib.

[0478] (8) Measurement of the prostate cancer (Du-145) activity of the compound

[0479] The drugs were the compound prepared in Example 31 (I-31), the commercially available CDK4 / 6 inhibitor palbociclib, and the first-line treatment for prostate cancer enzalutamide. The cell line was human prostate cancer Du-145 cells. The test animals were SPF-grade BALB / c male nude mice, 10 mice per group. The drug dosage settings are shown in Table 8.

[0480]

[0481] Drug manufacturing method:

[0482] Example 31 (100 mg / kg): 20 mg of the compound powder to be tested was weighed, dissolved in 2 mL of physiological saline, formulated into a 10 mg / mL drug, and administered orally by gastric tube feeding at a volume of 0.2 mL / 20 g.

[0483] Example 31 (200 mg / kg): 40 mg of the compound powder to be tested was weighed, dissolved in 2 ml of physiological saline, formulated into a 20 mg / mL drug, and administered orally by gastric nutrition at a volume of 0.2 mL / 20 g.

[0484] Enzalutamide (100 mg / kg): 20 mg of the compound powder to be tested was weighed and dissolved in 2 mL of physiological saline to prepare a 10 mg / mL drug for oral gastric tube administration in a volume of 0.2 mL / 20 g.

[0485] Experimental Methods: A nude mouse model of human prostate cancer xenograft was established by inoculating human prostate cancer Du-145 under the axillary skin of nude mice. Logarithmic-phase Du-145 cells were subcutaneously inoculated into the right axilla of 60 nude mice under sterile conditions, and the cell inoculation volume was 5 × 10⁶ 6 It was cell / mouse. The diameter of the xenograft was measured with a vernier caliper. The tumor was approximately 90 mm 3 Fifty nude mice with good growth status and uniform tumor size were selected for each group. Ten mice were randomly assigned to five groups: a model group, a low-dose group of Example 31 (100 mg / kg), a high-dose group of Example 31 (200 mg / kg), a positive drug palbociclib group (100 mg / kg), and a positive drug enzalutamide group (100 mg / kg). Test drugs Example 31, palbociclib, and enzalutamide were administered intragastrically once daily to the low-dose, high-dose, and positive drug groups. An equal amount of vehicle control was administered intragastrically to the model group. The antitumor effect of the test substances was observed dynamically by measuring tumor diameter. Tumor diameters were measured every other day, and the body weight of the nude mice was measured during the tumor diameter measurement. On day 35, the control group mice were sacrificed, and after surgical stripping, the tumor fragments were fixed in 10% formaldehyde and stored in liquid nitrogen for future use. The remaining mice were sacrificed on day 49, and after surgical stripping, the tumor fragments were fixed in 10% formaldehyde and stored in liquid nitrogen for later use.

[0486] The experimental results are illustrated in Fig. 3: The low-dose group (100 mg / kg) of test drug Example 31 showed superior tumor growth inhibition compared to the positive drug palbociclib (100 mg / kg) group, and the tumor growth inhibitory effect of the low-dose group (100 mg / kg) of test drug Example 31 and the positive drug enzalutamide (100 mg / kg) was similar. The high-dose group (200 mg / kg) of Example 31 significantly inhibited tumor growth compared to the positive drug palbociclib group (100 mg / kg) and the positive drug enzalutamide group (100 mg / kg), and the tumor volume began to decrease on day 31.

[0487] Therefore, the test drug prepared in Example 31 has a significant inhibitory effect on the growth of xenografts in nude mice with human prostate cancer Du-145, and the effect is superior to that of the positive control drug, the CDK4 / 6 inhibitor palbociclib, and the first-line treatment for prostate cancer enzalutamide.

Claims

Claim 1 Compound represented by the following chemical formula (I) or its pharmaceutically permissible salt: Here, X is C(O) or (CH2) n and; n is 0 or 1; R1 is hydrogen, a C1-C8 alkyl group or -NR4R5, where R4, R5 are hydrogen, a C1-C8 alkyl group or selected from C3-C8 cycloalkyl groups; R2 is F; and R3 is hydrogen or a C1-C8 alkyl group. Claim 2 In paragraph 1, the above X is C(O) or (CH2) n and; n is 0 or 1; and R1 is hydrogen, C1-C3 alkyl group or selected from -NR4R5, wherein R4 and R5 are selected from hydrogen, a C1-C3 alkyl group, cyclopentane, or cyclohexane; R2 is F; and R3 is hydrogen or a C1-C4 alkyl group, a compound represented by the formula (I) or a pharmaceutically permissible salt thereof. Claim 3 In paragraph 1, the above X is C(O) or (CH2) n and; n is 0 or 1; and R1 is hydrogen, C1-C3 alkyl group or selected from -NR4R5, wherein R4, R5 are selected from hydrogen, methyl group, ethyl group, cyclopentane or cyclohexane; said R2 is F and ; The above R3 is a compound represented by the formula (I) or a pharmaceutically acceptable salt thereof, wherein R3 is hydrogen or a C1-C4 alkyl group. Claim 4 In paragraph 1, the above X is (CH2) n or selected from C(O) , n is 0 or 1; and R1 is selected from hydrogen, a methyl group or -NR4R5. , Here, R4 is selected from the group consisting of hydrogen, methyl group, and ethyl group, and R5 is selected from hydrogen, methyl group, ethyl group, or cyclopentane; R2 is F; and R3 is A compound represented by the chemical formula (I) selected from hydrogen, ethyl group, and isopropyl group, or a pharmaceutically acceptable salt thereof. Claim 5 In claim 1, the compound represented by the formula (I) or a salt thereof that is pharmaceutically permissible, wherein the compound is selected from any one of the following compounds. Claim 6 In claim 1, the compound is a compound represented by the formula (I) or a salt thereof that is pharmaceutically permissible and selected from any one of the following compounds. Claim 7 In claim 1, the pharmaceutically permissible salt is an acidic addition salt of the compound represented by the formula (I), wherein the salt-forming acid comprises an inorganic acid and an organic acid, the inorganic acid comprises hydrochloric acid, sulfuric acid, phosphoric acid and methanesulfonic acid, and the organic acid comprises acetic acid, trichloroacetic acid, propionic acid, butyric acid, maleic acid, p-toluenesulfonic acid, malic acid, malonic acid, cinnamic acid, citric acid, fumaric acid, camphor acid, digluconic acid, aspartic acid and tartaric acid, the compound represented by the formula (I) or the pharmaceutically permissible salt thereof. Claim 8 In claim 1, the compound represented by formula (I) or a salt thereof that is pharmaceutically permissible, wherein the compound represented by formula (I) is prepared from compound (A) and compound (B) through a coupling reaction under the action of a palladium catalyst: Here, X is C(O) or (CH2) n and; n is 0 or 1; R1 is hydrogen, C1-C8 alkyl group, -NR4R5, where R4 and R5 are hydrogen, C1-C8 alkyl group or selected from C3-C8 cycloalkyl groups; R2 is F; and R3 is hydrogen, a C1-C8 alkyl group, a compound represented by the formula (I) or a pharmaceutically acceptable salt thereof. Claim 9 A pharmaceutical composition for treating and / or preventing at least one cancer or tumor-related disease selected from the group consisting of breast cancer, prostate cancer, lung cancer, multiple myeloma, leukemia, gastric cancer, ovarian cancer, colorectal cancer, liver cancer, pancreatic cancer, and human glioma, comprising a compound represented by the formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. Claim 10 A method for manufacturing a medicine for treating and / or preventing at least one cancer or tumor-related disease selected from the group consisting of breast cancer, prostate cancer, lung cancer, multiple myeloma, leukemia, gastric cancer, ovarian cancer, colorectal cancer, liver cancer, pancreatic cancer, and human glioma, wherein the medicine is manufactured using a compound represented by the chemical formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof.

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