Chromone spiro compound and preparation method and application thereof

Abstract

The application discloses a chromone spiro compound, a preparation method and application thereof, and the chromone spiro compound is shown as formula I. The compound has selective CDK4 / 6 inhibitory activity, can relieve and / or treat myelosuppression caused by chemotherapy, improve weight loss caused by chemotherapy and improve survival rate, and has good drug safety. The compound has simple preparation method, is convenient for large-scale production, and has great application prospect.

Description

Technical Field

[0001] This invention relates to a chromogenone spirocyclic compound, and also to the preparation and application of the above compound. Background Technology

[0002] Chemotherapy-induced myelosuppression (CIM) is a common adverse reaction during cancer chemotherapy, clinically manifested as neutropenia, lymphopenia, anemia, and thrombocytopenia. It can lead to adverse consequences including infection, sepsis, bleeding, and fatigue, and in severe cases, death. Current clinical treatment strategies for CIM are primarily symptomatic, mainly using growth factors such as recombinant human granulocyte colony-stimulating factor, erythropoietin, and thrombopoietin. However, these drugs target only a single lineage of blood cells, have a slow onset of action, and require long durations of treatment, potentially leading to bone marrow depletion and the risk of drug-related adverse reactions. Therefore, prophylactic use of a drug that can temporarily inhibit bone marrow cell proliferation can reduce the sensitivity of bone marrow cells to chemotherapy drugs, providing a new strategy for preventing or alleviating CIM.

[0003] Cyclin-dependent kinases (CDKs) play a crucial role in regulating the cell cycle. CDK4 / 6, belonging to the serine / threonine kinase family, bind to cyclin D in the pre-G1 phase of the cell cycle to form the CDK4 / 6-cyclin D complex. This complex promotes phosphorylation of retinoblastoma protein (RB), leading to the release of the transcription factor E2F and regulating the cell cycle transition from G1 to S phase. For hematopoietic stem and progenitor cells (HSPCs), CDK4 / 6 is essential for regulating HSPC proliferation.

[0004] CDK4 / 6 are key initiating proteins for the cell cycle transition from G1 to S phase. Inhibition of CDK4 / 6 can arrest the cell cycle in G1 phase, thereby reducing cell proliferation and playing an important role in the treatment of cancer and CIM. Currently, five selective CDK4 / 6 inhibitors, Palbociclib, Ribociclib, Abemaciclib, Dalpiciclib, and Trilaciclib (TC), have been approved for marketing. The first four are used for the treatment of breast cancer, while TC is the only one approved by the US FDA for the treatment of CIM in patients with advanced small cell lung cancer. Intravenous infusion of TC before chemotherapy can significantly induce G1 phase arrest in HSPCs, protecting bone marrow cells from the toxic damage of chemotherapy drugs.

[0005] Although TC is the only FDA-approved CDK4 / 6 inhibitor for the treatment of CIM, it still has some shortcomings, such as: (1) TC was initially developed as an anticancer drug, and its therapeutic effect on CIM was found in later clinical trials, but studies on drug toxicity and bone marrow penetration are lacking; (2) In clinical studies, TC can significantly increase neutrophil and erythrocyte counts, but has little effect on platelets, thus increasing the risk of bleeding in patients. Therefore, novel CDK4 / 6 inhibitors with novel structures, lower toxicity, better efficacy, and suitable for the treatment of CIM are still highly anticipated. Summary of the Invention

[0006] Purpose of the invention: The purpose of this invention is to provide a chromogenone spirocyclic compound with selective CDK4 / 6 inhibitory activity, and also to provide a method for preparing the above compound and its application in the preparation of CDK4 / 6 inhibitors, and in remedies for alleviating and / or treating chemotherapy-induced myelosuppression.

[0007] Technical solution: This invention discloses a chromogen ketone spirocyclic compound with the structure shown in Formula I:

[0008]

[0009] Where X is selected from O and (CH2). n C(O), NH or S(O)2, where n is 0 or 1;

[0010] R1 and R2 are selected from C3-C8 cycloalkyl, C3-C8 cycloalkyloxy or C3-C8 heterocyclic groups, wherein the C3-C8 cycloalkyl, C3-C8 cycloalkyloxy or C3-C8 heterocyclic group is optionally substituted by one or more substituents selected from halogen, hydroxyl, amino, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkyloxy or C4-C8 heterocyclic group;

[0011] R3 is hydrogen, halogen, hydroxyl, mercapto, amino, -OR', -SR', -NHR', or -NR'R", where R' and R" are C1-C3 alkyl, C2-C6 alkenyl, C3-C6 allyl, C2-C6 alkynyl, C1-C3 haloalkyl, C3-C6 cycloalkyl, C6-C 10 Aryl, C3-C 10 heteroaryl or C4-C8 heterocyclic group;

[0012] R4 is selected from C3-C8 cycloalkyloxy, C3-C8 heterocyclic, C3-C8 heterocyclic phenyl, or C3-C8 heterocyclic aryl, wherein the C3-C8 cycloalkyloxy, C3-C8 heterocyclic, C3-C8 heterocyclic phenyl, or C3-C8 heterocyclic aryl is optionally substituted by one or more substituents selected from halogen, hydroxyl, amino, nitro, cyano, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkyloxy, or C4-C8 heterocyclic.

[0013] Wherein, X is C(O) or (CH2). n n is 0 or 1; R1 and R2 are C3-C8 cycloalkyl or C3-C8 heterocyclic groups; R3 is hydrogen or halogen; R4 is a C3-C8 heterocyclic group. Wherein E is glutamic acid, and R1 and R2 are organic carbon chains selected from C6-C6. 24 C6-C substituted with alkyl or substituent groups 24 Alkyl, C6-C 24 C6-C substituents of alkenyl or substituent groups 24 alkenyl, C6-C 24 Alkyne or substituted C6-C 24 Alkyne group; n is a positive integer ≥ 1.

[0014] Wherein, X is C(O) or (CH2). n n is 0 or 1; R1 and R2 are C4-C6 cycloalkyl or C6 heterocyclic groups; R3 is a halogen; R4 is a C6 heterocyclic group.

[0015] Wherein, X is selected from: C(O) or (CH2). n n is 0 or 1; R1 and R2 are selected from cyclobutyl, cyclopentyl, cyclohexyl, cyclohexyl derivatives, tetrahydropyran, tetrahydrothiothioran or N-amide piperidine; R3 is F; R4 is selected from piperidine, piperidine derivatives, morpholine, piperazine or piperazine derivatives.

[0016] Specifically, the aforementioned chromogen spirocyclic compounds are selected from compounds 1-55:

[0017]

[0018]

[0019] This also includes pharmaceutically acceptable salts of the aforementioned chromogenone spirocyclic compounds, specifically, salts of compounds of general formula (I) formed by acid addition. The acids used for salt formation include inorganic and organic acids. The inorganic acids include hydrochloric acid, sulfuric acid, phosphoric acid, and methanesulfonic acid; the organic acids include galactoside, D-glucuronic acid, glycerophosphate, hippuric acid, hydroxyethanesulfonic acid, lactobionic acid, maleic acid, naphthalene-2-sulfonic acid, neopentanoic acid, and terephthalic acid. Cholic acid, benzenesulfonic acid, citric acid, lactic acid, malic acid, malonic acid, mandelic acid, acetic acid, propionic acid, tartaric acid, succinic acid, formic acid, methanesulfonic acid, picric acid, L-pyroglutamic acid, salicylic acid, gentian acid, p-toluenesulfonic acid, valeric acid, palmitic acid, stearic acid, lauric acid, adipic acid, 4-benzenesulfonic acid, fumaric acid, 3-hydroxynaphthalene-2-carboxylic acid, ascorbic acid, camphoric acid, camphorsulfonic acid, cinnamic acid, dichloroacetic acid, and ethanesulfonic acid.

[0020] The preparation method of the above-mentioned chromogen spirocyclic compound is as follows: (1) 1-(4-bromo-2-hydroxyphenyl) ethyl ketone as shown in Formula II is condensed with a cyclic ketone under alkaline conditions to obtain chromogen intermediate compound III; (2) chromogen intermediate compound III is reacted with dipinacol boronic acid ester under the action of palladium catalyst to obtain compound IV; (3) compound IV is coupled with a chloropyrimidine derivative to obtain compound V; (4) compound V is coupled with a 2-aminopyrimidine derivative under the action of palladium catalyst and phosphine ligand to obtain the chromogen spirocyclic compound shown in Formula I.

[0021] Specifically, the synthetic route is as follows: First, 1-(4-bromo-2-hydroxyphenyl)ethyl ketone (II) is reacted with cyclic ketones of different substitutions under alkaline conditions via Knoevenagel condensation to obtain the corresponding chromone intermediates of formula (III). Then, it is reacted with bis-pinacol boronic acid ester under palladium catalyst to obtain compound (IV). Subsequently, it undergoes a Suzuki coupling reaction with a chloropyrimidine derivative to obtain compound (V). Compound (V) is then reacted with a 2-aminopyrimidine derivative under palladium catalyst and phosphine ligand via a Buchwald-Hartwig coupling reaction to obtain compound (I). The synthetic route is as follows:

[0022]

[0023] In step (1), compound III is prepared from compound II. The base used is selected from pyrrolidine, piperidine, tetrahydropyridine, sodium hydroxide, potassium hydroxide, potassium carbonate, or sodium carbonate, preferably pyrrolidine. The reaction solvent is selected from methanol, ethanol, isopropanol, perfluoroisopropanol, n-butanol, or tert-butanol, preferably methanol. The reaction temperature is 50-100℃, preferably 60-80℃.

[0024] In step (2), compound IV is prepared from compound III. The base used is selected from potassium phosphate, potassium carbonate, sodium carbonate, cesium carbonate, potassium fluoride, potassium acetate, sodium acetate, potassium tert-butoxide, or sodium tert-butoxide, preferably potassium acetate. The palladium catalyst used is selected from Pd(dppf)Cl2, Pd(PPh3)2Cl2, Pd(PPh3)4, Pd2(dba)3, PdCl2(dippf)2, or [Pd(allyl)Cl]2, preferably Pd(dppf)Cl2. The reaction solvent is selected from dioxane, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, acetonitrile, or N-methylpyrrolidone, preferably dioxane. The reaction temperature is 50-100℃, preferably 80-100℃.

[0025] In step (3), compound V is prepared from compound IV. The base used is selected from sodium hydroxide, potassium hydroxide, barium hydroxide, potassium phosphate, potassium carbonate, sodium carbonate, cesium carbonate, potassium fluoride, potassium acetate, sodium acetate, sodium methoxide, potassium methoxide, sodium ethoxide, potassium ethoxide, potassium tert-butoxide, or sodium tert-butoxide, preferably sodium carbonate. The palladium catalyst used is selected from Pd(dppf)Cl2, Pd(PPh3)2Cl2, Pd(PPh3)4, Pd2(dba)3, PdCl2(dippf)2, or [Pd(allyl)Cl]2, preferably Pd(PPh3)2Cl2. The reaction solvent is selected from tetrahydrofuran, ethanol, acetonitrile, dioxane, ethylene glycol dimethyl ether, dimethyl sulfoxide, or toluene, preferably ethylene glycol dimethyl ether. The volume ratio of the reaction solvent to water is 8:1 to 12:1, preferably 10:1. The reaction temperature is 50-80℃, preferably 60-80℃.

[0026] In step (4), compound I is prepared from compound V. The base used is selected from sodium hydroxide, potassium hydroxide, potassium carbonate, sodium carbonate, cesium carbonate, sodium methoxide, potassium methoxide, sodium ethoxide, potassium ethoxide, potassium tert-butoxide, or sodium tert-butoxide, preferably cesium carbonate. The palladium catalyst used is selected from Pd2(dba)3 or Pd(OAc)2, preferably Pd2(dba)3. The phosphine ligand used is selected from PCy3, P(t-Bu)3, P(o-tolyl)3, DPPF, JohnPhos, SPhos, XantPhos, or BINAP, preferably BINAP. The reaction solvent is selected from toluene, xylene, dioxane, tetrahydrofuran, ethylene glycol dimethyl ether, N,N-dimethylformamide, N,N-dimethylacetamide, or dimethyl sulfoxide, preferably dioxane. The reaction temperature is 60-110℃, preferably 80-100℃.

[0027] The aforementioned chromogenone spirocyclic compounds can be used in the preparation of CDK4 / 6 inhibitors.

[0028] The aforementioned chromogenone spirocyclic compounds can also be used in the preparation of drugs to alleviate and / or treat chemotherapy-induced myelosuppression.

[0029] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof or an isomer thereof, and a pharmaceutically acceptable carrier.

[0030] Therapeutic effective amount can be an amount that, to some extent, alleviates one or more symptoms of a disease or condition in the subject, partially or completely restores one or more physiological or biochemical parameters related to or caused by the disease or condition to normal, and / or reduces the likelihood of the onset of the disease or condition; pharmaceutically acceptable carrier refers to an excipient or diluent that does not cause significant irritation to the organism and does not interfere with the biological activity and properties of the compound given.

[0031] Invention Principle: Currently, the only small molecule drug available for the clinical treatment of myelosuppression is triprazine, a selective CDK4 / 6 inhibitor. Structural analysis of triprazine and other marketed CDK4 / 6 inhibitors reveals that triprazine's myeloprotective effect stems from its unique azaspirocyclic skeleton. Therefore, this invention, based on the structure of triprazine, utilizes the principle of skeleton substitution, replacing the azaspirocyclic skeleton with a chromogenone spirocyclic skeleton while retaining the active fragment pyrimidine-NH-pyridine structure of the selective CDK4 / 6 inhibitor, thus synthesizing a series of chromogenone spirocyclic derivatives. These derivatives not only exhibit good CDK4 / 6 inhibitory activity but also excellent myeloprotective effects.

[0032] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: (1) The chromogen spirocyclic compound of the present invention has selective CDK4 / 6 inhibitory activity, and can alleviate and / or treat chemotherapy-induced bone marrow suppression, improve chemotherapy-induced weight loss and improve survival rate, and has good drug safety; (2) The synthesis steps of the chromogen spirocyclic compound of the present invention are short and the post-processing is simple, which facilitates large-scale production and has great application prospects. Attached Figure Description

[0033] Figure 1 The structural formula of the chromogenone spirocyclic compound of the present invention is shown below;

[0034] Figure 2 This is a schematic diagram illustrating the interaction between compound 42 and the CDK6 (PDB:5L2S) protein.

[0035] Figure 3Figure 42 shows the in vivo efficacy of compound 42. Figure A is a time-cycle diagram of the animal, Figures B to G show the effects of 42 on platelets, red blood cells, hemoglobin, white blood cells, lymphocytes, and neutrophils in the blood, Figure H shows the changes in body weight of mice after treatment with 5-FU or 42, and Figure I shows the survival curve of mice after 14 days of treatment with 5-FU or 42.

[0036] Figure 4 This is a schematic diagram of the safety evaluation of compound 42, where Figures A and B show the changes in food intake of female and male mice after administration of 42; Figures C and D show the organ coefficients of female and male mice after administration of 42.

[0037] Figure 5 The diagram shows the HE staining results, where Figure A shows the HE staining of various tissues from female mice, and Figure B shows the HE staining of various tissues from male mice. Detailed Implementation

[0038] The technical solution of the present invention will be further described below with reference to the embodiments. The test materials used in the embodiments can all be obtained through conventional means.

[0039] Example 1 7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,1'-cyclobutyl]-4-one (Compound 1)

[0040]

[0041] 4-Bromo-2-hydroxyacetophenone II (3 g, 23.2 mmol), cyclobutanone (1.96 g, 28 mmol), and pyrrolidine (2.5 g, 35 mmol) were dissolved in 50 mL of methanol. The mixture was stirred at 60 °C for 8 h, and the reaction was monitored by TLC until completion. The reaction solution was cooled to room temperature, poured into 100 mL of water, and extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed with water (80 mL) and saturated sodium chloride solution (80 mL), dried over anhydrous Na₂SO₄, and concentrated under reduced pressure to remove the solvent. The residue was purified by silica gel column chromatography (PE / EA, 40:1–25:1) to give compound III-1 as a pale yellow liquid (1.61 g, 43.1%). 1 H NMR (300MHz, CDCl3) δ7.71(d,J=8.4Hz,1H),7.20(d,J=1.8Hz,1H),7.14(dd,J=8.4,1.8Hz,1 H),2.90(s,2H),2.42-2.27(m,2H),2.12-2.13(m,2H),2.03-1.88(m,1H),1.81-1.65(m,1H).

[0042] Compound III-1 (1.54 g, 5.78 mmol), pinacol diboronate (1.76 g, 6.94 mmol), Pd(dppf)₂Cl₂ (0.21 g, 0.29 mmol), and KOAc (1.7 g, 17.34 mmol) were dissolved in 20 mL of dioxane. The mixture was stirred at 80 °C for 24 h under a nitrogen atmosphere, and the reaction was stopped by TLC. After cooling the reaction solution to room temperature, insoluble matter was removed by filtration. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE / EA, 10:1–6:1) to obtain compound IV-1 as a pale yellow liquid (1.55 g, 85.7%). 1 H NMR (300MHz, CDCl3) δ7.84(d,J=7.7Hz,1H),7.46(s,1H),7.41(dd,J=7.7,1.0Hz,1H),2.92(s, 2H),2.39-2.25(m,2H),2.21-2.13(m,2H),2.01-1.87(m,1H),1.82-1.64(m,1H),1.36(s,12H).

[0043] IV-1 (0.95 g, 3 mmol), 2,4-dichloro-5-fluoropyrimidine (0.75 g, 4.5 mmol), Pd(PPh3)2Cl2 (63 mg, 0.29 mmol), and Na2CO3 (0.65 g, 7.5 mmol) were dissolved in a mixture of ethylene glycol dimethyl ether (DME) and water (9 mL / 1 mL). The mixture was stirred at 80 °C for 12 h under a nitrogen atmosphere, and the reaction was stopped by TLC. After cooling the reaction solution to room temperature, the mixture was poured into 30 mL of water and extracted with ethyl acetate (20 mL × 3). The organic phases were combined, washed with water (60 mL) and saturated sodium chloride solution (60 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to remove the solvent. The residue was purified by silica gel column chromatography (PE / EA, 6:1–2:1) to give compound V-1 as a white solid (0.52 g, 53.3%). 1 H NMR(300MHz, CDCl3)δ8.60(d,J=2.9Hz,1H),8.00(d,J=8.2Hz,1H),7.82-7.80(m,1H),7.76(dt,J=8.3 ,1.5Hz,1H),2.98(s,2H),2.46-2.33(m,2H),2.28-2.17(m,2H),2.07-1.91(m,1H),1.84-1.72(m,1H). 13C NMR (151MHz, CDCl3) δ191.94,159.50,156.00,155.69,155.67,154.22,152.94,152.88,149.17,149 .00,138.39,138.35,127.24,122.82,121.45,121.41,119.64,119.59,81.39,46.02,33.07,12.16.

[0044] In a 25 mL sealed tube, V-1 (220 mg, 0.7 mmol), 5-(4-methylpiperazin-1-yl)-pyridine-2-amino (162 mg, 0.84 mmol), Pd2(dba)3 (64 mg, 0.07 mmol), BINAP (87 mg, 0.14 mmol), Cs2CO3 (456 mg, 1.4 mmol), and dioxane (8 mL) were added sequentially. The mixture was stirred at 100 °C for 12 h under a nitrogen atmosphere, and the reaction was stopped by TLC. After cooling the reaction solution to room temperature, the solvent was removed by concentration under reduced pressure. The residue was purified by silica gel column chromatography (DCM / CH3OH, 40:1–10:1) to give a yellow solid compound 1 (135 mg, 41.4%). 1 H NMR (300MHz, CDCl3) δ8.47(d,J=3.3Hz,1H),8.33(s,1H),8.27(d,J=9.1Hz,1H),8.09(d,J=3.0Hz,1H),8.00(d,J=8.6H z,1H),7.76-7.72(m,2H),7.39(dd,J=9.1,3.0Hz,1H),3.24(t,J=5.0Hz,4H),2.99(s,2H),2.67(t,J=5.0Hz,4H),2.49 -2.35(m,5H),2.29-2.17(m,2H),2.06-1.93(m,2H). 13 C NMR (101MHz, CDCl3) δ192.27,159.49,155.67,155.64,152.08,150.48,150.39,149.53,147.81,147.55,146.19,142.94,140.71,1 40.66,136.83,127.03,126.52,122.11,121.43,121.38,119.35,119.28,112.37,81.21,54.94,49.63,46.10,46.05,33.12,12.16.

[0045] Example 2 7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,1'-cyclopentyl]-4-one (Compound 2)

[0046] Compound 2 was prepared using the same method as Compound 1; 1 H NMR (300MHz, CDCl3) δ8.48-8.44(m,2H),8.27(d,J=9.0Hz,1H),8.10(d,J=3.0 Hz,1H),8.01(d,J=8.2Hz,1H),7.72(dt,J=8.2,1.4Hz,1H),7.69-7.67(m,1H), 7.38(dd,J=9.1,3.0Hz,1H),3.22(t,J=5.0Hz,4H),2.91(s,2H),2.65(t,J=5.0 Hz,4H),2.41(s,3H),2.22-2.10(m,2H),1.98-1.90(m,2H),1.83-1.65(m,4H). 13 C NMR (101MHz, CDCl3) δ192.33,160.23,155.71,155.68,152.04,150.62,150.53,149.50,147.74,147.48,146.24,142.95,140.56,1 40.50,136.81,127.00,126.50,122.07,121.05,120.99,119.46,119.38,112.39,90.37,54.97,49.69,47.08,46.16,37.50,23.90.

[0047] Example 3 7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,1'-cyclohexyl]-4-one (Compound 3)

[0048] Compound 3 was prepared using the same method as compound 1; 1H NMR (300MHz, CDCl3) δ8.47(d,J=3.3Hz,1H),8.36(s,1H),8.27(d,J=9.1Hz,1H),8.09(d,J=3.0Hz,1H),8.02-7.97(m,1H),7.73-7.69(m,2H),7.37(dd ,J=9.1,3.0Hz,1H),3.23(t,J=5.0Hz,4H),2.78(s,2H),2.66(t,J=5.0Hz, 4H),2.41(s,3H),2.12-1.95(m,3H),1.85-1.64(m,3H),1.62-1.49(m,4H). 13 C NMR (101MHz, CDCl3) δ192.35,159.45,155.66,152.08,150.69,150.60,149.54,147.71,147.44,146.16,142.99,140.66,140.60,1 36.81,126.77,126.45,121.98,120.97,120.91,119.35,119.27,112.40,80.45,54.96,49.68,48.23,46.14,34.75,25.15,21.45.

[0049] Example 4 7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)-4'-methylspiro[benzopyran-2,1'-cyclohexyl]-4-one (Compound 4)

[0050] Compound 4 was prepared using the same method as compound 1; 1 H NMR (300MHz, CDCl3) δ8.47(d,J=3.3Hz,1H),8.33(s,1H),8.29(d,J=9.1Hz,1 H),8.09(d,J=2.9Hz,1H),8.00(d,J=8.1Hz,1H),7.76-7.68(m,2H),7.37(dd ,J=9.1,3.0Hz,1H),3.25(t,J=5.0Hz,4H),2.74(s,2H),2.71(t,J=5.0Hz,4H ),2.44(s,3H),2.20-2.12(m,2H),1.67-1.37(m,7H),0.99(d,J=3.1Hz,3H). 13CNMR(101MHz, CDCl3)δ192.43,159.38,155.67,155.64,152.06,150.65,150.56,149.51,147.74,147.48,146.20,142.96,140.69,140.63, 136.82,126.79,126.40,122.05,121.04,120.98,119.33,119.26,11 2.40,79.60,54.96,49.67,49.12,46.14,34.44,31.60,29.59,22.24.

[0051] Example 5 4'-Ethyl-7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,1'-cyclohexyl]-4-one (Compound 5)

[0052] Compound 5 was prepared using the same method as compound 1; 1 H NMR (300MHz, CDCl3) δ8.47(d,J=3.3Hz,1H),8.33-8.27(m,2H),8.09(d,J=3.3Hz,1H),8. 00(d,J=8.2Hz,1H),7.74(s,1H),7.71(dt,J=8.1,1.4Hz,1H),7.37(dd,J=9.1,3.0Hz,1H ),3.24(t,J=5.0Hz,4H),2.75(s,2H),2.68(t,J=4.9Hz,4H),2.43(s,3H),2.19(d,J=9.5 Hz,2H),1.67-1.57(m,2H),1.50-1.39(m,4H),1.37-1.19(m,3H),0.93(t,J=7.3Hz,3H). 13 C NMR (101MHz, CDCl3) δ192.45,159.40,155.68,155.65,152.04,150.61,150.52,149.49,147.78,147.52,146.25,142.92,140.70,140.65,136 .81,126.79,126.41,122.06,121.05,120.99,119.31,119.24,112.39, 79.97,54.96,49.65,49.15,46.13,38.26,34.41,29.57,27.26,11.52.

[0053] Example 6 4'-tert-butyl-7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,1'-cyclohexyl]-4-one (Compound 6)

[0054] Compound 6 was prepared using the same method as compound 1; 1 H NMR(300MHz, CDCl3) δ8.46(d,J=3.3Hz,1H),8.29(d,J=9.7Hz,1H),8.23(s,1H),8.08 (d,J=3.8Hz,1H),8.00(d,J=7.4Hz,1H),7.74-7.68(m,2H),7.36(dd,J=9.1,3.0Hz,1 H),3.24(t,J=5.0Hz,4H),2.74(s,2H),2.69(t,J=5.1Hz,4H),2.43(s,3H),2.24(d,J =12.9Hz,2H),1.68-1.58(m,2H),1.57-1.35(m,4H),1.14-1.03(m,1H),0.92(s,9H). 13 C NMR (101MHz, CDCl3) δ192.46,159.40,155.64,155.61,152.06,150.65,150.56,149.52,147.77,147.51,146.15,142.96,140.69,140.63,136 .80,126.80,126.40,122.07,121.10,121.03,119.17,119.11,112.29, 79.62,54.95,49.65,49.14,47.05,46.13,34.96,32.48,27.57,21.99.

[0055] Example 7 7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)-4',4'-dimethylspiro[benzopyran-2,1'-cyclohexyl]-4-one (Compound 7)

[0056] Compound 7 was prepared using the same method as compound 1; 1H NMR(300MHz, CDCl3)δ8.46(d,J=3.3Hz,1H),8.28(d,J=9.1Hz,1H),8.24(s,1H),8 .08(d,J=3.0Hz,1H),8.00(d,J=8.0Hz,1H),7.74-7.69(m,2H),7.37(dd,J=9.1,3. 0Hz,1H),3.24(t,J=5.0Hz,4H),2.79(s,2H),2.68(t,J=5.0Hz,4H),2.43(s,3H),2 .03-1.96(m,2H),1.72-1.59(m,4H),1.30-1.24(m,2H),1.02(s,3H),0.97(s,3H). 13 C NMR (126MHz, CDCl3) δ192.30,159.48,155.62,151.91,150.71,150.63,149.87,147.72,147.50,146.17,142.99,140.68,136.94, 126.82,126.46,122.08,121.06,121.02,119.31,119.25,112.40,80.26,54.93,49.60,48.13,46.04,34.20,30.96,29.43,25.37.

[0057] Example 8 4',4'-Difluoro-7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,1'-cyclohexyl]-4-one (Compound 8)

[0058] Compound 8 was prepared using the same method as Compound 1; 1 H NMR (300MHz, DMSO-d6) δ9.86 (s, 1H), 8.70 (d, J = 3.4Hz, 1H), 8.02 (s, 1H), 8. 00(d,J=6.5Hz,1H),7.92(d,J=8.2Hz,1H),7.75(t,J=1.2Hz,1H),7.71(dt,J =8.2,1.4Hz,1H),7.41(dd,J=9.1,3.1Hz,1H),3.12(t,J=5.0Hz,4H),2.97(s ,2H),2.50-2.45(m,4H),2.24(s,3H),2.19-1.94(m,6H),1.86-1.72(m,2H). 13CNMR(126MHz,DMSO-d6)δ191.66,158.80,156.43,151.80,150.67,149.78,148.55,145.92,143.25,140.96, 135.95,126.99,125.51,122.09,121.91,119.25,113.72,78.94,54.97,48.93,46.83,46.26,30.79,30.72.

[0059] Example 9 7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,1'-cyclohexyl-4',2”-[1,3]-dioxolane]-4-one (Compound 9)

[0060] Compound 9 was prepared using the same method as compound 1; 1 H NMR (300MHz, CDCl3) δ8.47(d,J=3.3Hz,1H),8.27(d,J=9.0Hz,1H),8.25(s,1H),8.08(d ,J=2.9Hz,1H),8.02(d,J=8.6Hz,1H),7.77-7.72(m,2H),7.39(dd,J=9.1,3.0Hz,1H),4 .04-3.95(m,4H),3.25(t,J=5.0Hz,4H),2.80(s,2H),2.69(t,J=5.0Hz,4H),2.44(s,3H ),2.18(d,J=13.8Hz,2H),2.11-1.98(m,2H),1.87-1.74(m,2H),1.66(d,J=13.0Hz,2H). 13 C NMR (101MHz, CDCl3) δ191.88,159.19,155.67,155.64,152.04,150.35,150.26,149.49,147.90,147.63,146.21,142.92,140.90,140.85,136. 75,126.90,126.48,121.93,121.32,121.27,119.34,119.26,112.47,1 07.91,79.20,64.47,64.34,54.94,49.58,47.97,46.10,32.21,30.01.

[0061] Example 10 7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)-5”,5”-dimethylspiro[benzopyran-2,1'-cyclohexyl-4',2”-[1,3]-dioxane]-4-one (Compound 10)

[0062] Compound 10 was prepared using the same method as Compound 1; 1 H NMR (300MHz, CDCl3) δ8.47(d,J=3.3Hz,1H),8.26(d,J=9.1Hz,1H),8.21(s,1H),8.08 (d,J=3.0Hz,1H),8.01(d,J=8.6Hz,1H),7.76-7.72(m,2H),7.38(dd,J=9.1,3.0Hz,1H ),3.57(s,2H),3.50(s,2H),3.25(t,J=5.1Hz,4H),2.78(s,2H),2.69(t,J=5.0Hz,4H ),2.43(s,3H),2.16-2.03(m,4H),1.94-1.83(m,2H),1.79-1.66(m,2H),1.00(s,6H). 13 C NMR (101MHz, CDCl3) δ191.91,159.21,155.68,155.65,152.03,150.42,1 50.33,149.49,147.84,147.59,146.20,142.93,140.86,140.81,136.76 ,126.89,126.47,121.89,121.24,121.19,119.36,119.28,112.44,96.7 5,79.63,70.13,54.95,49.61,48.02,46.12,30.92,30.25,27.38,22.70.

[0063] Example 11 7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)-2',3',5',6'-tetrahydrospiro[benzopyran-2,4'-pyran]-4-one (Compound 11)

[0064] Compound 11 was prepared using the same method as Compound 1; 1H NMR (300MHz, CDCl3) δ8.50-8.47(m,2H),8.26(d,J=9.1Hz,1H),8.11(d,J=2.4Hz,1H),8.02(dd,J=7.8,0.9Hz,1H),7.78-7.72(m,2H),7.37(dd,J= 9.1,3.0Hz,1H),3.97-3.78(m,4H),3.23(t,J=5.0Hz,4H),2.83(s,2H),2 .66(t,J=5.0Hz,4H),2.41(s,3H),2.09-2.00(m,2H),1.90-1.77(m,2H). 13 C NMR (101MHz, CDCl3) δ191.35,158.93,155.69,155.66,152.06,150.42,150.33,149.51,147.82,147.56,146.12,142.99,140.97,1 40.92,136.81,126.92,126.46,121.88,121.52,121.46,119.26,119.18,112.40,77.59,63.15,54.94,49.62,48.31,46.10,34.79.

[0065] Example 12 7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)-2',3',5',6'-tetrahydrospiro[benzopyran-2,4'-thiopyran]-4-one (Compound 12)

[0066] Compound 12 was prepared using the same method as compound 1; 1 H NMR(300MHz, CDCl3)δ8.49(d,J=3.3Hz,1H),8.41(s,1H),8.26(d,J=9.2Hz, 1H),8.10(d,J=2.3Hz,1H),8.01(d,J=8.7Hz,1H),7.77-7.73(m,2H),7.38( dd,J=9.1,3.0Hz,1H),3.24(t,J=5.0Hz,4H),3.20-3.07(m,2H),2.78(s,2H ),2.67(t,J=5.0Hz,4H),2.52-2.43(m,4H),2.42(s,3H),1.95-1.81(m,2H). 13CNMR(126MHz, CDCl3)δ191.28,158.65,155.62,151.93,150.46,150.39,149.89,147.80,147.59,146.14,142.98,140.98,1 37.00,126.93,126.51,121.87,121.55,121.50,119.24,119.18,112.45,78.52,54.88,49.50,48.89,45.93,35.66,23.51.

[0067] Example 13: tert-butyl-7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)-4-oxaspiro[benzopyran-2,4'-piperidine]-1'-carboxylate (Compound 13)

[0068] Compound 13 was prepared using the same method as compound 1; 1 H NMR (300MHz, CDCl3) δ8.47(d,J=3.2Hz,1H),8.28-8.23(m,2H),8.08(d,J=2.3Hz,1H),8.02(dd,J=7.9,0.8Hz,1H),7.77-7.72(m,2H),7.37(dd,J=9.1 ,3.0Hz,1H),3.92(s,2H),3.36-3.17(m,6H),2.80(s,2H),2.67(t,J=5.0Hz ,4H),2.42(s,3H),2.09(d,J=13.7Hz,2H),1.72-1.61(m,2H),1.49(s,9H). 13 C NMR (101MHz, CDCl3) δ191.26,158.86,155.68,155.65,154.72,152.05,150.39,150.31,149.51,147.81,147.55,146.10,142.99,140.99,1 40.94,136.75,126.96,126.47,121.86,121.55,121.50,119.22,119 .14,112.39,79.88,78.32,54.93,49.62,48.08,46.11,34.00,28.43.

[0069] Example 14 1'-Acetyl-7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 14)

[0070] Compound 14 was prepared using the same method as compound 1; 1 H NMR(300MHz, CDCl3)δ8.48(d,J=3.3Hz,1H),8.29-8.22(m,2H),8.08(d,J=2.9Hz,1 H),8.03(d,J=8.1Hz,1H),7.80-7.74(m,2H),7.37(dd,J=9.1,3.0Hz,1H),4.45(d,J =13.7Hz,1H),3.73-3.52(m,2H),3.23(t,J=5.0Hz,4H),3.15(t,J=11.5Hz,1H),2.8 1(s,2H),2.68(t,J=5.0Hz,4H),2.42(s,3H),2.22-2.11(m,5H),1.76-1.60(m,2H). 13 C NMR (101MHz, CDCl3) δ190.93,168.97,158.67,155.73,155.70,152.02,1 50.29,150.21,149.47,147.85,147.59,146.13,142.99,141.11,141.06, 136.74,127.02,126.46,121.77,121.73,121.68,119.15,119.08,112.4 2,78.12,54.93,49.61,48.01,46.11,41.85,36.88,34.56,33.86,21.46.

[0071] Example 15 1'-Acetyl-7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)-1'-(methanesulfonyl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 15)

[0072] Compound 15 was prepared using the same method as Compound 1; 1H NMR(300MHz, CDCl3)δ8.48(d,J=3.2Hz,1H),8.31(s,1H),8.25(d,J=8.4Hz,1H), 8.08(d,J=2.3Hz,1H),8.04(d,J=8.2Hz,1H),7.80-7.73(m,2H),7.37(dd,J=9.1 ,3.0Hz,1H),3.69(d,J=11.9Hz,2H),3.26-3.12(m,6H),2.87(s,3H),2.83(s,2H ),2.66(t,J=4.9Hz,4H),2.42(s,3H),2.24(d,J=13.9Hz,2H),1.93-1.82(m,2H). 13 C NMR (101MHz, CDCl3) δ190.63,158.41,155.71,155.68,152.06,150.32,150.23,149.51,147.78,147.52,145.99,143.08,141.09,141. 04,136.67,127.14,126.42,121.91,121.85,121.82,119.01,118.94,112.40,77.21,54.94,49.60,47.93,46.14,41.35,35.12,33.84.

[0073] Example 16 7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)-1'-propionylspiro[benzopyran-2,4'-piperidin]-4-one (Compound 16)

[0074] Compound 16 was prepared using the same method as compound 1; 1 H NMR (300MHz, CDCl3) δ8.48 (d, J = 3.3Hz, 1H), 8.31 (s, 1H), 8.25 (d, J = 9.1Hz, 1H), 8.08 (d, J = 2. 9Hz,1H),8.03(d,J=8.1Hz,1H),7.79-7.74(m,2H),7.37(dd,J=9.1,3.0Hz,1H),4.46(d,J=13. 6Hz,1H),3.71(d,J=13.8Hz,1H),3.54(t,J=12.6Hz,1H),3.26-3.10(m,5H),2.81(s,2H),2.69 -2.62(m,4H),2.45-2.34(m,5H),2.21-2.09(m,2H),1.73-1.59(m,2H),1.18(t,J=7.4Hz,3H).13 C NMR (101MHz, CDCl3) δ190.99,172.29,158.71,155.71,155.68,152.03,15 0.31,150.22,149.48,147.85,147.59,146.12,142.98,141.09,141.04,13 6.75,127.01,126.47,121.78,121.71,121.65,119.16,119.09,112.41,78 .24,54.92,49.60,48.03,46.10,40.89,37.04,34.59,33.96,26.54,9.54.

[0075] Example 17 1'-Butyryl-7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 17)

[0076] Compound 17 was prepared using the same method as compound 1; 1 H NMR (300MHz, CDCl3) δ8.47(d,J=3.3Hz,1H),8.24(d,J=9.1Hz,1H),8.18(s,1H),8.07(d,J=2.3Hz, 1H),8.03(d,J=8.7Hz,1H),7.80-7.74(m,2H),7.37(dd,J=9.1,3.0Hz,1H),4.46(d,J=13.4Hz,1H) ,3.72(d,J=13.6Hz,1H),3.55(t,J=12.7Hz,1H),3.27-3.09(m,5H),2.81(s,2H),2.67(t,J=5.0Hz ,4H),2.42(s,3H),2.38-2.30(m,2H),2.18-2.10(m,2H),1.74-1.62(m,4H),1.00(t,J=7.4Hz,3H). 13C NMR (101MHz, CDCl3) δ190.97,171.51,158.71,155.70,155.67,152.05,150. 33,150.24,149.51,147.84,147.58,146.10,142.99,141.08,141.02,136.80 ,127.02,126.45,121.80,121.71,121.65,119.16,119.09,112.39,78.25,54.92,49.59,48.04,46.08,41.07,36.96,35.29,34.70,33.98,18.80,14.04.

[0077] Example 18 1'-(cyclopropylacyl)-7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 18)

[0078] Compound 18 was prepared using the same method as compound 1; 1 H NMR (300MHz, CDCl3) δ8.47(d,J=3.2Hz,1H),8.26(d,J=9.7Hz,1H),8.19(s,1H),8.07(d,J=2.3H z,1H),8.04(d,J=8.8Hz,1H),7.79-7.75(m,2H),7.37(dd,J=9.1,3.0Hz,1H),4.43(d,J=13.2Hz, 1H),4.06(d,J=13.5Hz,1H),3.67(d,J=13.1Hz,1H),3.33-3.12(m,5H),2.83(s,2H),2.77-2.66( m,4H),2.46(s,3H),2.20-2.09(m,2H),1.83-1.63(m,3H),1.05-0.98(m,2H),0.83-0.75(m,2H). 13CNMR(101MHz,CDCl3)δ191.02,172.00,158.75,155.72,155.69,152.04,15 0.33,150.24,149.50,147.85,147.59,146.10,143.01,141.08,141.03,13 6.76,127.01,126.44,121.81,121.69,121.63,119.19,119.11,112.40,78 .41,54.94,49.62,48.07,46.12,41.02,37.66,34.74,33.96,10.98,7.47.

[0079] Example 19 7-(5-fluoro-2-((5-(4-methylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)-1'-(2,2,2-trifluoroacetyl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 19)

[0080]

[0081] Compound 19 was prepared using the same method as compound 1; 1 H NMR (300MHz, CDCl3) δ8.48(d,J=3.3Hz,1H),8.24(d,J=9.1Hz,1H),8.20(s,1H),8.08(d ,J=2.4Hz,1H),8.05(d,J=8.3Hz,1H),7.83–7.76(m,2H),7.37(dd,J=9.1,3.0Hz,1H),4. 45(d,J=13.4Hz,1H),3.92(d,J=14.1Hz,1H),3.67(t,J=13.2Hz,1H),3.39-3.20(m,5H), 2.84(s,2H),2.75-2.66(m,4H),2.44(s,3H),2.26(d,J=14.1Hz,2H),1.83-1.68(m,2H). 13C NMR (126MHz, CDCl3) δ190.33,158.40,155.71,151.90,150.17,149.86,147.85,147.64,145.99,143.11,141.24,141.20,136.86,127. 19,126.39,122.07,122.02,121.78,119.07,119.02,117.65,112.42,77.67,54.94,49.59,47.94,46.07,41.22,39.02,34.66,33.69.

[0082] Example 20 1'-Acetyl-7-(5-fluoro-2-((5-(piperidin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 20)

[0083] In a 25 mL sealed tube, V-14 (220 mg, 0.56 mmol), 5-(piperidin-1-yl)pyridine-2-amino (120 mg, 0.672 mmol), Pd2(dba)3 (52 mg, 0.056 mmol), BINAP (70 mg, 0.112 mmol), Cs2CO3 (365 mg, 1.12 mmol), and dioxane (8 mL) were added sequentially. The mixture was stirred at 100 °C for 12 h under a nitrogen atmosphere, and the reaction was stopped by TLC. After cooling the reaction solution to room temperature, the solvent was removed by concentration under reduced pressure. The residue was purified by silica gel column chromatography (DCM / CH3OH, 40:1–10:1) to give a yellow solid compound 19 (139 mg, 46.7%). 1 H NMR (300MHz, CDCl3) δ8.47(d,J=3.2Hz,1H),8.33(s,1H),8.24(d,J=8.1Hz,1H),8 .06(d,J=2.8Hz,1H),8.03(dd,J=8.1,0.6Hz,1H),7.80-7.74(m,2H),7.40(dd,J= 9.1,3.0Hz,1H),4.45(d,J=13.5Hz,1H),3.75-3.52(m,2H),3.21-3.09(m,5H),2. 81(s,2H),2.22-2.15(m,2H),2.14(s,3H),1.81-1.72(m,4H),1.71-1.56(m,4H). 13C NMR (101MHz, CDCl3) δ190.92,168.93,158.67,155.76,155.73,152.00,152.24,152.15,149.45,147.87,147.61,145.66,144.01,141.15,141 .09,136.89,127.01,126.97,121.75,121.69,119.16,119.08,112.43, 78.12,51.23,48.03,41.85,36.88,34.55,33.88,25.76,24.01,21.46.

[0084] Example 21 1'-Acetyl-7-(5-fluoro-2-((5-(4-methylpiperidin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 21)

[0085] Compound 21 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-methylpiperidin-1-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.47(d,J=3.3Hz,1H),8.32(s,1H),8.25(d,J=9.1Hz,1H),8.05(s,1H),8.03(d,J= 8.1Hz,1H),7.80-7.75(m,2H),7.42(dd,J=9.1,3.0Hz,1H),4.45(d,J=13.5Hz,1H),3.74-3.51(m,4H),3 .22-3.08(m,1H),2.81(s,2H),2.74(td,J=11.9,2.5Hz,2H),2.22-2.16(m,2H),2.14(s,3H),1.80(d,J= 12.7Hz,2H),1.73-1.60(m,2H),1.58-1.49(m,1H),1.43(td,J=12.1,3.8Hz,2H),1.02(d,J=6.2Hz,3H). 13C NMR (101MHz, CDCl3) δ190.94,168.98,158.66,155.74,155.72,152.00,1 50.24,150.15,149.45,147.86,147.60,145.59,143.72,141.14,141.09 ,136.88,127.01,126.92,121.75,121.69,119.16,119.08,112.46,78.1 1,50.56,48.01,41.86,36.89,34.54,33.98,33.88,30.45,21.89,21.45.

[0086] Example 22 1'-Acetyl-7-(5-fluoro-2-((5-(4-methoxypiperidin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 22)

[0087] Compound 22 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-methoxypiperidin-1-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.47(d,J=3.2Hz,1H),8.34(s,1H),8.27(d,J=9.1Hz,1H),8.05(s,1H), 8.04-8.02(m,1H),7.80-7.75(m,2H),7.44(dd,J=9.1,2.3Hz,1H),4.45(d,J=13.6Hz,1H),3.7 4-3.52(m,2H),3.51-3.42(m,3H),3.41(s,3H),3.22-3.09(m,1H),3.02-2.94(m,2H),2.81(s, 2H),2.23-2.15(m,2H),2.15(s,3H),2.07-2.04(m,1H),1.85-1.72(m,3H),1.71-1.61(m,2H). 13C NMR (101MHz, CDCl3) δ190.92,168.94,158.67,155.72,155.70,152.02,150 .27,150.19,149.47,147.87,147.60,145.86,143.21,141.11,141.06,137 .04,127.02,126.96,121.77,121.74,121.68,119.16,119.08,112.40,78.13,75.55,55.67,48.01,47.75,41.85,36.88,34.56,33.87,30.50,21.46.

[0088] Example 23 Ethyl 1-(6-((4-(1'-acetyl-4-oxospiro[benzopyran-2,4'-piperidine]-7-one)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)piperidine-4-carboxylate (Compound 23)

[0089] Compound 23 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with ethyl 1-(6-aminopyridin-3-yl)piperidin-4-carboxylate, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.48(d,J=3.2Hz,1H),8.40(s,1H),8.26(d,J=9.1Hz,1H),8.06(d,J=2.9Hz,1H), 8.03(d,J=8.2Hz,1H),7.81–7.73(m,2H),7.40(dd,J=9.1,3.0Hz,1H),4.45(d,J=13.6Hz,1H),4.19(q,J =7.1Hz,2H),3.73-3.53(m,4H),3.15(t,J=12.3Hz,1H),2.88-2.77(m,4H),2.51-2.41(m,1H),2.22-2. 15(m,2H),2.14(s,3H),2.08-2.04(m,1H),2.00-1.85(m,3H),1.74-1.61(m,2H),1.30(t,J=7.1Hz,3H). 13C NMR (101MHz, CDCl3) δ190.91,174.70,168.94,158.67,155.69,155.66,152.05,15 0.31,150.22,149.50,147.86,147.60,146.00,143.32,141.08,141.03,137.04,1 27.19,127.03,121.78,121.73,121.68,119.16,119.09,112.43,78.14,60.55,51.83,49.81,48.01,41.85,40.70,40.58,36.88,34.57,33.87,28.00,21.46,14.26.

[0090] Example 24 1'-Acetyl-7-(5-fluoro-2-((5-(4-dimethylaminopiperidin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 24)

[0091] Compound 24 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-dimethylaminopiperidin-1-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.47(d,J=3.3Hz,1H),8.23(d,J=9.1Hz,1H),8.07(s,1H),8.06-8.02( m,2H),7.80-7.74(m,2H),7.38(dd,J=9.1,3.0Hz,1H),4.45(d,J=13.4Hz,1H),3.67(d,J=11 .2Hz,3H),3.64-3.51(m,1H),3.15(t,J=12.2Hz,1H),2.81(s,2H),2.75(dd,J=12.2,2.4Hz, 2H),2.40(s,6H),2.16-2.21(m,2H),2.15(s,3H),2.03(d,J=12.4Hz,2H),1.61-1.80(m,5H). 13C NMR (101MHz, CDCl3) δ190.93,168.97,158.68,155.69,155.66,152.05,150 .29,150.20,149.50,147.85,147.59,145.82,143.25,141.10,141.05,137 .11,127.03,126.88,121.78,121.74,121.68,119.15,119.08,112.36,78.13,61.86,49.80,48.02,41.86,41.64,36.89,34.56,33.88,28.17,21.46.

[0092] Example 25 1'-Acetyl-7-(5-fluoro-2-((5-(4-morpholinopyridin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 25)

[0093] Compound 25 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-morpholinoperidin-1-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.47(d,J=3.3Hz,1H),8.23(d,J=9.1Hz,1H),8.10(s,1H),8.07-8. 01(m,2H),7.80-7.74(m,2H),7.37(dd,J=9.1,3.0Hz,1H),4.45(d,J=13.5Hz,1H),3.82( s,4H),3.68(d,J=12.3Hz,3H),3.64-3.52(m,1H),3.15(t,J=12.3Hz,1H),2.81(s,2H),2 .81-2.59(m,6H),2.25-2.15(m,2H),2.15(s,3H),2.09-2.00(m,2H),1.78-1.64(m,5H). 13C NMR (101MHz, CDCl3) δ190.92,168.96,158.67,155.72,155.69,152.02,150. 27,150.18,149.47,147.87,147.61,145.88,143.21,141.11,141.05,137.07 ,127.02,126.84,121.77,121.73,121.67,119.16,119.08,112.38,78.13,67.22,61.74,49.79,49.76,48.01,41.85,36.88,34.55,33.87,27.99,21.46.

[0094] Example 26 1'-Acetyl-7-(5-fluoro-2-((5-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 26)

[0095] Compound 26 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.47(d,J=3.3Hz,1H),8.25-8.19(m,2H),8.07(d,J=2.9Hz,1H),8.03(d,J=8 .1Hz,1H),7.80-7.73(m,2H),7.36(dd,J=9.1,3.0Hz,1H),4.44(d,J=13.5Hz,1H),3.67(d,J=12.0H z,3H),3.63-3.51(m,1H),3.15(t,J=12.0Hz,1H),2.81(s,2H),2.80-2.64(m,6H),2.54(s,4H),2. 43-2.37(m,1H),2.34(s,3H),2.19(s,2H),2.14(s,3H),1.99(d,J=12.4Hz,2H),1.78-1.60(m,4H). 13C NMR (101MHz, CDCl3) δ190.91,168.94,158.66,155.73,155.70,152.01,150.26 ,150.17,149.46,147.86,147.60,145.79,143.18,141.11,141.06,137.03,12 7.01,126.77,121.76,121.73,121.67,119.15,119.07,112.36,78.12,61.46,55.42,49.90,49.03,48.01,46.02,41.85,36.88,34.55,33.87,28.09,21.46.

[0096] Example 27 4-(6-((4-(1'-acetyl-4-oxospiro[benzopyran-2,4'-piperidine]-7-one)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)piperidine-1-carboxylic acid tert-butyl ester (Compound 27)

[0097] Compound 27 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 1-(6-aminopyridin-3-yl)piperidin-4-carboxylic acid tert-butyl ester, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.57(s,1H),8.52(d,J=3.2Hz,1H),8.36(d,J=8.6Hz,1H),8.21(d,J=2 .3Hz,1H),8.04(dd,J=8.1,0.6Hz,1H),7.80-7.75(m,2H),7.62(dd,J=8.8,2.4Hz,1H),4.45 (d,J=13.5Hz,1H),4.27(s,2H),3.75-3.52(m,2H),3.21-3.08(m,1H),2.92-2.80(m,4H),2. 75-2.64(m,1H),2.22-2.12(m,5H),1.86(d,J=13.0Hz,2H),1.75-1.62(m,4H),1.51(s,9H). 13C NMR (101MHz, CDCl3) δ190.89,168.97,158.69,155.50,155.47,154.83,152 .30,151.19,150.48,150.39,149.74,147.89,147.63,146.40,140.87,140. 82,136.53,134.98,127.08,121.87,121.73,121.68,1119.18,119.11,111.85,79.62,78.18,48.01,41.85,39.64,36.89,34.60,33.84,28.50,21.46.

[0098] Example 28 1'-Acetyl-7-(5-fluoro-2-((5-(piperidin-4-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 28)

[0099] Compound 28 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(piperidin-4-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, DMSO-d6) δ10.12(s,1H),9.20(s,1H),8.76(d,J=3.3Hz,1H),8.20(d,J=2.3Hz,1H) ,8.15(d,J=8.6Hz,1H),7.95-7.90(m,1H),7.75-7.69(m,2H),7.65(dd,J=8.7,2.5Hz,1H),4.12( d,J=13.3Hz,1H),3.73-3.62(m,1H),3.46-3.38(m,2H),3.32(brs,2H),3.07-2.94(m,3H),2.94( s,2H),2.90-2.81(m,1H),2.02(s,3H),1.98-1.86(m,5H),1.84-1.72(m,1H),1.69-1.55(m,1H). 13C NMR(101MHz,DMSO-d6)δ191.73,168.76,158.93,156.13,152.09,152.07,150.00,149.93,148.81,148.59,146.64,140.73,140.68,13 6.33,134.27,126.99,122.15,121.71,119.27,119.22,112.76,79.18,47.30,43.87,41.83,36.92,36.48,34.28,33.48,29.52,21.79.

[0100] Example 29 1'-Acetyl-7-(5-fluoro-2-((5-(1-methylpiperidin-4-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 29)

[0101] Compound 29 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(1-methylpiperidin-4-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.51(d,J=3.2Hz,1H),8.38(s,1H),8.32(d,J=8.2Hz,1H),8.25(d,J=2. 4Hz,1H),8.04(dd,J=8.0,0.7Hz,1H),7.81-7.74(m,2H),7.63(dd,J=8.6,2.5Hz,1H),4.45(d, J=13.6Hz,1H),3.76-3.50(m,2H),3.22-3.02(m,3H),2.82(s,2H),2.59-2.48(m,1H),2.40(s, 3H),2.22-2.16(m,2H),2.15(s,3H),2.13-2.09(m,1H),1.95-1.83(m,5H),1.76-1.60(m,2H). 13C NMR (101MHz, CDCl3) δ190.90,168.98,158.68,155.66,155.63,152.19,151 .23,150.39,150.30,149.64,147.90,147.64,146.81,140.98,140.92,136 .39,135.35,127.06,121.81,121.76,121.70,119.15,119.08,111.81,78.15,56.17,48.01,46.42,41.86,38.90,36.89,34.58,33.85,33.32,21.45.

[0102] Example 30 1'-Acetyl-7-(5-fluoro-2-((5-morpholinpyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 30)

[0103] Compound 30 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-morpholinopyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.48(d,J=3.2Hz,1H),8.43(s,1H),8.31(dd,J=9.1,0.7Hz,1H),8.07-8.00(m,2H),7.80-7.75(m,2H),7.39(dd,J=9.2,3 .0Hz,1H),4.44(d,J=13.7Hz,1H),3.95-3.87(m,4H),3.74-3.51(m,2H) ,3.22-3.07(m,5H),2.82(s,2H),2.22-2.10(m,5H),1.76-1.60(m,2H). 13 C NMR (125MHz, CDCl3) δ190.86,168.97,158.73,155.55,151.98,150.47,150.39,149.93,147.79,147.58,146.14,143.09,141.00,140.96, 136.91,127.08,126.46,121.90,121.77,121.72,119.18,119.12,11 2.59,78.20,66.80,49.86,48.06,41.89,36.93,34.64,33.92,21.43.

[0104] Example 31 4-(6-((4-(1'-acetyl-4-oxospiro[benzopyran-2,4'-piperidine]-7-one)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)piperazine-1-carboxylic acid tert-butyl ester (Compound 31)

[0105] Compound 31 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 4-(6-aminopyridin-3-yl)piperazine-1-carboxylic acid tert-butyl ester, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.48(d,J=3.2Hz,1H),8.39(s,1H),8.31(d,J=9.1Hz,1H),8.07-8.00(m,2H),7.81-7.74(m,2H),7.41(dd,J=9.2,3 .0Hz,1H),4.44(d,J=13.7Hz,1H),3.71-3.55(m,6H),3.21-3.07(m,5H),2.82(s,2H),2.22-2.10(m,5H),1.74-1.64(m,2H),1.51(s,9H). 13 C NMR (101MHz, CDCl3) δ190.92,168.99,158.68,155.63,155.60,154.66,152 .09,150.36,150.27,149.54,147.87,147.61,146.55,142.96,141.01,140 .96,137.08,127.33,127.04,121.80,121.72,121.67,119.16,119.09,112.47,80.09,78.16,49.99,47.99,41.85,36.89,34.57,33.84,28.44,21.45.

[0106] Example 32 1'-Acetyl-7-(5-fluoro-2-((5-(piperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 32)

[0107] Compound 32 was prepared by replacing 5-(piperazin-1-yl)pyridine-2-amino with 5-(piperazin-1-yl)pyridine-2-amino, following the same method as compound 20. 1H NMR (400MHz, CDCl3) δ8.47(d,J=3.2Hz,1H),8.26-8.22(m,1H),8.09(s,1H),8.06-8.01(m,2H),7.80-7.74(m,2H),7.36(dd,J=9.1,3.0Hz ,1H),4.45(d,J=13.3Hz,1H),3.72-3.54(m,2H),3.23-3.12(m,5H),3.11-3.07(m,4H),2.82(s,2H),2.20-2.13(m,5H),1.70-1.63(m,5H). 13 C NMR (126MHz, DMSO-d6) δ191.72,168.71,158.94,156.38,151.83,149.88,149.81,148.74,148.54,146.33,143.11,140.90,140.85 ,136.33,126.94,126.01,122.12,121.72,119.23,119.18,113.64,79.18,48.50,47.31,44.65,41.82,36.91,34.28,33.50,21.78.

[0108] Example 33 1'-Acetyl-7-(5-fluoro-2-((5-(4-ethylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 33)

[0109] Compound 33 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-ethylpiperazin-1-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR(300MHz, CDCl3)δ8.48(d,J=3.3Hz,1H),8.28-8.22(m,2H),8.10-8.07(m,1H),8.03( d,J=8.3Hz,1H),7.80-7.74(m,2H),7.37(dd,J=9.1,3.0Hz,1H),4.45(d,J=13.3Hz,1H),3 .74-3.52(m,2H),3.26(t,J=4.9Hz,4H),3.21-3.09(m,1H),2.81(s,2H),2.73(t,J=4.5Hz ,4H),2.58(q,J=7.2Hz,2H),2.21-2.12(m,5H),1.75-1.61(m,2H),1.20(t,J=7.2Hz,3H). 13C NMR (101MHz, CDCl3) δ190.92,168.94,158.67,155.75,155.72,152.01,1 50.28,150.19,149.46,147.87,147.60,146.12,143.05,141.13,141.07 ,136.74,127.02,126.39,121.76,121.68,119.15,119.08,112.41,78.1 3,52.66,52.37,49.66,48.02,41.85,36.88,34.56,33.87,21.46,11.97.

[0110] Example 34 1'-Acetyl-7-(5-fluoro-2-((5-(4-isopropylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 34)

[0111] Compound 34 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-isopropylpiperazin-1-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.47(d,J=3.2Hz,1H),8.24(d,J=8.8Hz,1H),8.21(s,1H),8.07(d, J=2.3Hz,1H),8.03(d,J=8.2Hz,1H),7.81-7.73(m,2H),7.37(dd,J=9.1,3.0Hz,1H),4.4 5(d,J=13.6Hz,1H),3.75-3.51(m,2H),3.23(t,J=4.9Hz,4H),3.20-3.10(m,1H),2.81(s ,2H),2.77(t,J=5.1Hz,4H),2.21-2.10(m,5H),1.77-1.61(m,3H),1.15(d,J=6.5Hz,6H). 13C NMR (101MHz, CDCl3) δ190.93,168.96,158.67,155.74,155.71,152.01,1 50.27,150.18,149.46,147.86,147.60,146.05,143.13,141.13,141.08 ,136.72,127.01,126.37,121.75,121.69,119.15,119.07,112.41,78.1 2,54.59,50.00,48.57,48.02,41.86,36.89,34.56,33.87,21.46,18.57.

[0112] Example 35 1'-Acetyl-7-(5-fluoro-2-((5-(4-(2-alkoxyethyl)piperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 35)

[0113] Compound 35 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-(2-alkoxyethyl)piperazin-1-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR(300MHz, CDCl3)δ8.48(d,J=3.3Hz,1H),8.38(s,1H),8.24(d,J=9.2Hz,1H),8.08 (d,J=2.3Hz,1H),8.03(d,J=8.3Hz,1H),7.80-7.73(m,2H),7.36(dd,J=9.1,3.0Hz,1H ),4.44(d,J=13.4Hz,1H),3.73-3.52(m,4H),3.40(s,3H),3.24(t,J=5.0Hz,4H),3.2 1-3.08(m,1H),2.81(s,2H),2.76-2.67(m,6H),2.21-2.10(m,5H),1.75-1.61(m,2H). 13C NMR (101MHz, CDCl3) δ190.94,168.97,158.67,155.73,155.70,152.02,15 0.27,150.18,149.47,147.87,147.61,146.08,143.06,141.12,141.07,13 6.74,127.02,126.40,121.76,121.68,119.15,119.07,112.41,78.12,70. 05,58.98,57.94,53.41,49.51,48.01,41.85,36.89,34.56,33.86,21.45.

[0114] Example 36 1'-Acetyl-7-(5-fluoro-2-((5-(4-cyclopropylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 36)

[0115] Compound 36 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-cyclopropylpiperazin-1-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.48(d,J=3.3Hz,1H),8.43(s,1H),8.24(d,J=9.3Hz,1H),8.09(d,J=2 .3Hz,1H),8.03(d,J=7.9Hz,1H),7.80-7.73(m,2H),7.36(dd,J=9.1,3.0Hz,1H),4.44(d,J= 13.6Hz,1H),3.74-3.52(m,2H),3.17(t,J=5.2Hz,4H),3.12-3.10(m,1H),2.85(t,J=5.0Hz, 4H), 2.81 (s, 2H), 2.19 (s, 1H), 2.17-2.10 (m, 5H), 1.74-1.65 (m, 2H), 0.54 (d, J = 6.2Hz, 4H). 13C NMR (101MHz, CDCl3) δ190.92,168.95,158.67,155.73,155.70,152.02,15 0.29,150.20,149.48,147.85,147.59,146.06,143.17,141.11,141.06,13 6.81,127.03,126.48,121.77,121.74,121.68,119.15,119.08,112.39,78 .13,53.15,49.67,48.02,41.86,38.52,36.88,34.56,33.87,21.46,5.81.

[0116] Example 37 1'-Acetyl-7-(5-fluoro-2-((5-(4-cyclohexylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 37)

[0117] Compound 37 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-cyclohexylpiperazin-1-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.47(d,J=3.3Hz,1H),8.24(d,J=9.1Hz,1H),8.09(s,1H),8.06(d,J=2.9Hz ,1H),8.03(dd,J=8.1,0.6Hz,1H),7.80-7.74(m,2H),7.37(dd,J=9.1,3.0Hz,1H),4.45(d,J=13.5 Hz,1H),3.75-3.51(m,2H),3.29(brs,4H),3.15(t,J=12.2Hz,1H),2.89(brs,4H),2.81(s,2H),2 .12-2.10(m,5H),2.03(brs,2H),1.88(brs,2H),1.75-1.61(m,5H),1.38-1.22(d,J=13.3Hz,4H). 13C NMR (101MHz, CDCl3) δ190.94,168.98,158.67,155.70,155.67,152.04,150. 28,150.19,149.49,147.85,147.59,145.96,143.17,141.11,141.06,136.6 8,127.02,126.34,121.77,121.69,119.14,119.07,112.40,78.12,63.59,50.09,48.84,48.02,41.86,36.89,34.55,33.88,28.91,26.27,25.84,21.45.

[0118] Example 38 1'-Acetyl-7-(5-fluoro-2-((5-(4-(oxecyclobutane-3-yl)piperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 38)

[0119] Compound 38 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-(oxecyclobutan-3-yl)piperazin-1-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.48(d,J=3.2Hz,1H),8.42(s,1H),8.28(d,J=9.3Hz,1H),8.07(d, J=2.9Hz,1H),8.03(dd,J=8.1,0.6Hz,1H),7.80-7.74(m,2H),7.38(dd,J=9.1,3.0Hz,1H) ,4.79-4.66(m,4H),4.44(d,J=13.4Hz,1H),3.75-3.51(m,3H),3.25(t,J=5.0Hz,4H),3.2 0-3.10(m,1H),2.81(s,2H),2.58(t,J=5.0Hz,4H),2.20-2.12(m,5H),1.76-1.60(m,2H). 13C NMR (101MHz, CDCl3) δ190.91,168.96,158.68,155.65,155.63,152.09,150 .34,150.25,149.54,147.85,147.59,146.25,142.89,141.05,141.00,136 .77,127.04,126.67,121.80,121.73,121.67,119.16,119.09,112.44,78.15,75.36,59.19,49.50,49.42,48.02,41.85,36.88,34.57,33.86,21.46.

[0120] Example 39 1'-Acetyl-7-(5-fluoro-2-((5-(4-cyclopropylmethylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 39)

[0121] Compound 39 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-cyclopropylmethylpiperazin-1-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR(300MHz, CDCl3)δ8.48(d,J=3.3Hz,1H),8.29-8.22(m,2H),8.08(dd,J=3.0,0.7Hz,1H),8.03 (dd,J=8.1,0.6Hz,1H),7.80-7.74(m,2H),7.37(dd,J=9.1,3.0Hz,1H),4.45(d,J=13.6Hz,1H),3. 74-3.53(m,2H),3.26(t,J=5.0Hz,4H),3.22-3.08(m,1H),2.83-2.74(m,6H),2.40(d,J=6.6Hz,2H ),2.21-2.13(m,5H),1.74-1.62(m,2H),1.02-0.91(m,1H),0.64-0.56(m,2H),0.23-0.16(m,2H). 13C NMR (101MHz, CDCl3) δ190.93,168.97,158.67,155.74,155.71,152.02,15 0.28,150.19,149.47,147.87,147.61,146.09,143.10,141.12,141.07,1 36.74,127.02,126.42,121.76,121.69,119.15,119.07,112.42,78.12,6 3.70,53.05,49.61,48.01,41.86,36.89,34.56,33.87,21.45,8.25,3.98.

[0122] Example 40 1'-Acetyl-7-(5-fluoro-2-((5-(4-acetylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 40)

[0123] Compound 40 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-acetylpiperazin-1-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.49(d,J=3.2Hz,1H),8.46(s,1H),8.32(d,J=9.1Hz,1H ),8.07-8.01(m,2H),7.79-7.75(m,2H),7.41(dd,J=9.2,3.0Hz,1H),4.44(d, J=13.7Hz,1H),3.86-3.79(m,2H),3.72-3.63(m,3H),3.65-3.52(m,1H),3.20 -3.11(m,5H),2.82(s,2H),2.22-2.16(m,5H),2.14(s,3H),1.76-1.61(m,2H). 13C NMR (101MHz, CDCl3) δ190.92,169.06,169.00,158.68,155.59,155.56,152.14 ,150.41,150.32,149.58,147.85,147.59,146.80,142.62,140.98,140.93,13 7.24,127.41,127.05,121.82,121.72,121.66,119.18,119.11,112.48,78.18,50.35,49.96,47.99,46.16,41.84,41.26,36.88,34.59,33.82,21.46,21.37.

[0124] Example 41 1'-Acetyl-7-(5-fluoro-2-((5-(4-trifluoroformylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 41)

[0125] Compound 41 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-trifluoroformylpiperazin-1-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.59(s,1H),8.50(d,J=3.2Hz,1H),8.37(d,J=9.1Hz,1H),8 .07-8.01(m,2H),7.80-7.74(m,2H),7.43(dd,J=9.2,2.9Hz,1H),4.44(d,J=13.6H z,1H),3.90(t,J=5.1Hz,2H),3.83(t,J=5.0Hz,2H),3.74-3.52(m,2H),3.23(t,J= 5.1Hz,4H),3.19-3.08(m,1H),2.82(s,2H),2.22-2.12(m,5H),1.76-1.61(m,2H). 13C NMR (101MHz, CDCl3) δ190.88,168.98,158.69,155.71,155.51,155.48,155.35,1 52.20,150.48,150.39,149.65,147.86,147.59,147.19,142.11,140.90,140.85 ,137.43,127.75,127.07,121.86,121.71,121.65,119.20,119.12,117.83,112.47,78.20,50.38,49.88,47.99,45.69,43.17,41.84,36.88,34.62,33.80,21.47.

[0126] Example 42 1'-Acetyl-7-(5-fluoro-2-((5-(4-cyclopropanoylpiperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 42)

[0127] Compound 42 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-trifluoroformylpiperazin-1-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.50 (s, 1H), 8.49 (s, 1H), 8.33 (d, J = 9.1Hz, 1H), 8.07-8.0 1(m,2H),7.81-7.74(m,2H),7.43(dd,J=9.2,3.0Hz,1H),4.44(d,J=13.6Hz,1H), 3.87(brs,4H),3.73-3.53(m,2H),3.27-3.09(m,5H),2.82(s,2H),2.22-2.10(m, 5H),1.84-1.77(m,1H),1.75-1.64(m,2H),1.09-1.01(m,2H),0.89-0.80(m,2H). 13C NMR (101MHz, CDCl3) δ190.93,172.16,169.02,158.68,155.61,155.58,152.12 ,150.38,150.29,149.56,147.87,147.61,146.70,142.69,141.00,140.95,13 7.09, 127.30, 127.05, 121.81, 121.72, 121.67, 119.18, 119.10, 112.50, 78.17, 50.42, 49.92, 47.99, 45.38, 41.85, 36.89, 34.58, 33.82, 21.46, 10.98, 7.64.

[0128] Example 43 4-(6-((4-(1'-acetyl-4-oxospiro[benzopyran-2,4'-piperidine]-7-one)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)piperazine-1-carboxylic acid ethyl ester (Compound 43)

[0129] Compound 43 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with ethyl 4-(6-aminopyridin-3-yl)piperazine-1-carboxylate, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.49(d,J=3.2Hz,1H),8.43(s,1H),8.30(d,J=9.1Hz,1 H),8.08-8.01(m,2H),7.80-7.74(m,2H),7.40(dd,J=9.1,3.0Hz,1H),4.44( d,J=13.3Hz,1H),4.20(q,J=7.1Hz,2H),3.75-3.52(m,6H),3.22-3.07(m,5H) ),2.82(s,2H),2.22-2.10(m,5H),1.76-1.62(m,2H),1.31(t,J=7.1Hz,3H). 13C NMR (101MHz, CDCl3) δ190.90,168.97,158.68,155.63,155.60,155.45,152. 11,150.39,150.30,149.55,147.85,147.59,146.67,142.90,141.01,140.95 ,137.27,127.39,127.05,121.81,121.72,121.67,119.17,119.09,112.44,78.17,61.61,50.01,48.00,43.56,41.85,36.88,34.59,33.84,21.46,14.71.

[0130] Example 44 1'-Acetyl-7-(5-fluoro-2-((5-(morpholin-4-carbonyl)piperazin-1-yl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 44)

[0131] Compound 44 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(morpholino-4-carbonyl)piperazin-1-yl)pyridine-2-amino, following the same method as compound 20. 1 H NMR(300MHz, CDCl3)δ9.05(s,1H),8.60-8.53(m,2H),8.45(dd,J=2.3,0.8Hz,1H),8.08-8.03(m,1H),7.92(dd,J=8.8,2.3Hz,1H),7.81-7.76(m,2 H),4.44(d,J=13.4Hz,1H),3.81-3.65(m,8H),3.59(t,J=11.5Hz,2H),3. 14(t,J=12.7Hz,1H),2.82(s,2H),2.22-2.12(m,5H),1.75-1.62(m,2H). 13 C NMR (101MHz, CDCl3) δ190.87,169.01,168.09,158.73,155.06,153.76,152.60,150.76,150.67,150.03,148.00,147.74,147.35,140.55,1 40.50,138.11,127.20,124.61,121.99,121.71,121.65,119.20,119 .13,111.24,78.25,66.88,47.99,41.85,36.89,34.66,33.76,21.45.

[0132] Example 45 4-(6-((4-(1'-acetyl-4-oxospiro[benzopyran-2,4'-piperidine]-7-one)-5-fluoropyrimidin-2-yl)amino)nicotinyl)piperazine-1-carboxylic acid tert-butyl ester (Compound 45)

[0133] Compound 45 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 4-(6-aminonicotinyl)piperazine-1-carboxylic acid tert-butyl ester, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.98(s,1H),8.58(d,J=3.1Hz,1H),8.53-8.48(m,2H),8.05(d,J=8.2Hz,1H),7.87(d,J=8.5Hz,1H),7.81-7.75(m,2H),4 .44(d,J=13.4Hz,1H),3.81-3.56(m,6H),3.51(s,4H),3.14(t,J=12.7H z,1H),2.82(s,2H),2.21-2.10(m,5H),1.76-1.63(m,2H),1.50(s,9H). 13 C NMR (101MHz, CDCl3) δ190.85,169.00,168.31,158.73,155.11,155.07,154 .54,153.82,152.58,150.73,150.64,150.01,148.03,147.77,147.44,140. 56,140.51,138.00,127.19,124.75,121.99,121.71,121.65,119.20,119.13,111.19,80.51,78.25,48.00,41.85,36.89,34.66,33.76,28.38,21.46.

[0134] Example 46 1'-Acetyl-7-(5-fluoro-2-((5-(4-methylpiperazin-1-carbonyl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidine]-4-one (Compound 46)

[0135] Compound 46 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-methylpiperazin-1-carbonyl)pyridine-2-amino, following the same method as compound 20. 1H NMR (300MHz, CDCl3) δ8.73 (s, 1H), 8.57 (d, J = 3.2Hz, 1H), 8.52-8.45 (m, 2H), 8.05(dd,J=7.9,0.8Hz,1H),7.85(dd,J=8.6,2.4Hz,1H),7.78(dd,J=8.1,1. 2Hz,2H),4.44(d,J=13.6Hz,1H),3.91-3.52(m,6H),3.14(t,J=12.4Hz,1H), 2.82(s,2H),2.55(s,4H),2.41(s,3H),2.23-2.12(m,5H),1.76-1.62(m,2H). 13 C NMR (101MHz, CDCl3) δ190.88,169.01,168.05,158.73,155.17,155.14,153.70,152.54,150.68,150.59,149.97,148.03,147.76,147.50,140. 62,140.56,137.92,127.18,125.07,121.96,121.71,121.65,119.19,1 19.12,111.11,78.23,48.00,46.00,41.85,36.89,34.65,33.76,21.45.

[0136] Example 47 1'-Acetyl-7-(5-fluoro-2-((5-(4-ethylpiperazin-1-carbonyl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidine]-4-one (Compound 47)

[0137] Compound 47 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-ethylpiperazine-1-carbonyl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.55 (d, J = 3.1Hz, 1H), 8.51-8.47 (m, 2H), 8.47-8.44 (m, 1H),8.07-8.03(m,1H),7.85(dd,J=8.8,2.2Hz,1H),7.78(dd,J=8.0,1.2Hz,2H ),4.45(d,J=13.6Hz,1H),3.95-3.52(m,6H),3.14(t,J=12.6Hz,1H),2.82(s, 2H),2.58(s,4H),2.23-2.11(m,5H),1.74-1.63(m,4H),1.18(t,J=7.2Hz,3H). 13C NMR (101MHz, CDCl3) δ190.89,169.02,167.97,158.73,155.17,155.14,1 53.68,152.54,150.67,150.58,149.97,148.03,147.76,147.51,140.62, 140.57,137.94,127.18,125.10,121.96,121.71,121.65,119.19,119.12 ,111.10,78.23,52.26,48.00,41.86,36.90,34.65,33.76,21.45,11.86.

[0138] Example 48 1'-Acetyl-7-(5-fluoro-2-((5-(4-acetylpiperazin-1-carbonyl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidine]-4-one (Compound 48)

[0139] Compound 48 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-acetylpiperazin-1-carbonyl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.95(s,1H),8.58(d,J=3.1Hz,1H),8.54-8.49(m,2H),8.07-8.02(m,1H),7.87(dd,J=8.6,2.4Hz,1H),7.78(dd,J=8.1,1.2Hz,2 H),4.44(d,J=13.6Hz,1H),3.81-3.61(m,7H),3.59-3.53(m,3H),3.21-3. 07(m,1H),2.82(s,2H),2.23-2.15(m,5H),2.14(s,3H),1.76-1.62(m,2H). 13 C NMR (101MHz, CDCl3) δ190.86,169.29,169.02,168.39,158.73,155.08,155.0 5,153.99,152.60,150.77,150.68,150.03,148.03,147.77,147.51,140.53, 140.48,138.06,127.19,124.38,121.99,121.69,121.64,119.21,119.14,111.24,78.26,47.98,46.15,41.84,41.50,36.89,34.65,33.74,21.46,21.42.

[0140] Example 49 1'-Acetyl-7-(5-fluoro-2-((5-(4-cyclopropionylpiperazin-1-carbonyl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 49)

[0141] Compound 49 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(4-cyclopropylpiperazin-1-carbonyl)pyridine-2-amino, following the same method as compound 20. 1 H NMR (300MHz, CDCl3) δ8.88(s,1H),8.57(d,J=3.1Hz,1H),8.54(d,J=8.9Hz,1H),8.48(d, J=2.2Hz,1H),8.05(d,J=8.8Hz,1H),7.90(dd,J=8.6,2.1Hz,1H),7.82-7.75(m,2H),4.4 4(d,J=13.5Hz,1H),3.88-3.65(m,8H),3.59(t,J=12.4Hz,2H),3.14(t,J=12.7Hz,1H),2 .83(s,2H),2.23-2.21(m,5H),1.78-1.63(m,3H),1.08-1.02(m,2H),0.89-0.80(m,2H). 13 C NMR (101MHz, CDCl3) δ190.86,172.44,169.01,168.37,158.73,155.09,155 .06,153.94,152.60,150.76,150.67,150.03,148.03,147.77,147.51,140 .54,140.49,138.07,127.19,124.50,121.99,121.71,121.65,119.21,119.14,111.23,78.26,47.99,41.85,36.89,34.66,33.75,21.46,11.06,7.82.

[0142] Example 50 1'-Acetyl-7-(5-fluoro-2-((5-(morpholinomethyl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 50)

[0143] Compound 50 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-(morpholinomethyl)pyridine-2-amino, following the same method as compound 20. 1H NMR(300MHz, CDCl3)δ8.53(d,J=3.2Hz,1H),8.51(s,1H),8.38(d,J=8.6Hz, 1H),8.31-8.29(m,1H),8.05(d,J=7.9Hz,1H),7.82-7.75(m,3H),4.45(d,J= 13.6Hz,1H),3.75(t,J=4.6Hz,4H),3.69-3.56(m,2H),3.54(s,2H),3.24-3 .09(m,1H),2.82(s,2H),2.52(s,4H),2.23-2.11(m,5H),1.75-1.63(m,2H). 13 C NMR (101MHz, CDCl3) δ190.90,169.00,158.70,155.53,155.50,152.29,1 52.11,150.46,150.37,149.73,148.80,147.94,147.68,140.90,140.84, 139.24,127.09,126.81,121.86,121.74,121.68,119.17,119.10,111.5 5,78.19,66.90,60.21,53.45,48.01,41.85,36.89,34.58,33.84,21.46.

[0144] Example 51 4-((6-((4-(1'-acetyl-4-oxospiro[benzopyran-2,4'-piperidine]-7-one)-5-fluoropyrimidin-2-yl)amino)pyridin-3-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (Compound 51)

[0145] Compound 51 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 4-((6-aminopyridin-3-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester, following the same method as compound 20. 1H NMR (300MHz, CDCl3) δ8.53(d,J=3.2Hz,1H),8.44(s,1H),8.37(d,J=8.6Hz,1H), 8.28(d,J=2.2Hz,1H),8.04(dd,J=8.1,0.6Hz,1H),7.81-7.72(m,3H),4.45(d,J =13.7Hz,1H),3.74-3.57(m,2H),3.54(s,2H),3.47(s,4H),3.15(t,J=11.6Hz,1 H),2.82(s,2H),2.45(s,4H),2.23-2.10(m,5H),1.69-1.60(m,2H),1.47(s,9H). 13 C NMR (101MHz, CDCl3) δ190.89,168.98,158.70,155.52,155.49,154.74,152.29 ,152.09,150.45,150.36,149.73,148.75,147.96,147.69,140.89,140.84,13 9.18, 127.09, 126.94, 121.86, 121.74, 121.68, 119.17, 119.10, 111.54, 79.71, 78.19, 59.83, 52.73, 48.01, 41.85, 36.89, 34.59, 33.84, 28.43, 26.91, 21.46.

[0146] Example 52 1'-Acetyl-7-(5-fluoro-2-((5-(((4-methylpiperazin-1-yl)methyl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 52)

[0147] Compound 52 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-((4-methylpiperazin-1-yl)methyl)pyridine-2-amino, following the same method as compound 20. 1H NMR(300MHz, CDCl3)δ8.52(d,J=3.2Hz,1H),8.43(s,1H),8.36(dd,J=8.5,0.8Hz,1H) ,8.30-8.27(m,1H),8.04(dd,J=8.0,0.6Hz,1H),7.81-7.75(m,2H),7.72(dd,J=8.6, 2.3Hz,1H),4.45(d,J=13.3Hz,1H),3.73-3.57(m,2H),3.52(s,2H),3.15(t,J=12.2H z,1H),2.82(s,2H),2.56(s,8H),2.36(s,3H),2.21-2.13(m,5H),1.75-1.63(m,2H). 13 C NMR (101MHz, CDCl3) δ190.90,168.96,158.70,155.56,155.53,152.26,151 .98,150.39,150.30,149.70,148.78,147.97,147.71,140.93,140.88,139 .17,127.31,127.08,121.85,121.75,121.69,119.16,119.08,111.49,78.18,59.79,55.02,52.85,48.01,45.94,41.86,36.89,34.56,33.86,21.46.

[0148] Example 53 1'-Acetyl-7-(5-fluoro-2-((5-(((4-ethylpiperazin-1-yl)methyl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 53)

[0149] Compound 53 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-((4-ethylpiperazin-1-yl)methyl)pyridine-2-amino, following the same method as compound 20. 1H NMR (300MHz, CDCl3) δ8.56 (s, 1H), 8.53 (d, J = 3.2Hz, 1H), 8.35 (dd, J = 8.5, 0.8Hz, 1H), 8.2 9(dd,J=2.3,0.8Hz,1H),8.04(dd,J=8.1,0.6Hz,1H),7.81-7.75(m,2H),7.72(dd,J=8.6,2 .3Hz,1H),4.45(d,J=13.4Hz,1H),3.74-3.57(m,2H),3.52(s,2H),3.23-3.09(m,1H),2.8 2(s,2H),2.75-2.31(m,10H),2.21-2.13(m,5H),1.75-1.62(m,2H),1.12(t,J=7.2Hz,3H). 13 C NMR (101MHz, CDCl3) δ190.90,168.95,158.70,155.56,155.53,152.26,151. 97,150.38,150.29,149.70,148.80,147.97,147.71,140.93,140.88,139.19 ,127.27,127.07,121.85,121.75,121.69,119.16,119.08,111.49,78.18,59.82,52.80,52.69,52.29,48.02,41.86,36.89,34.56,33.86,21.46,11.89.

[0150] Example 54 1'-Acetyl-7-(5-fluoro-2-((5-(((4-acetylpiperazin-1-yl)methyl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 54)

[0151] Compound 54 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-((4-acetylpiperazin-1-yl)methyl)pyridine-2-amino, following the same method as compound 20. 1H NMR (300MHz, CDCl3) δ8.56-8.52(m,2H),8.38(dd,J=8.5,0.8Hz,1H),8.29(dd ,J=2.3,0.9Hz,1H),8.04(dd,J=8.1,0.6Hz,1H),7.82-7.71(m,3H),4.45(d,J =13.7Hz,1H),3.74-3.58(m,4H),3.55-3.46(m,4H),3.22-3.09(m,1H),2.82( s,2H),2.53-2.42(m,4H),2.22-2.12(m,5H),2.11(s,3H),1.73-1.61(m,2H). 13 C NMR (101MHz, CDCl3) δ190.90,169.01,158.70,155.49,155.46,152.31,152.18 ,150.49,150.40,149.75,148.70,147.95,147.69,140.86,140.80,139.15,12 7.10, 126.70, 121.87, 121.73, 121.67, 119.18, 119.11, 111.59, 78.20, 59.66, 52.96, 52.54, 48.00, 46.21, 41.85, 41.34, 36.89, 34.58, 33.83, 21.46, 21.36.

[0152] Example 55 1'-Acetyl-7-(5-fluoro-2-((5-(((4-cyclopropionylpiperazin-1-yl)methyl)pyridin-2-yl)amino)pyrimidin-4-yl)spiro[benzopyran-2,4'-piperidin]-4-one (Compound 55)

[0153] Compound 55 was prepared by replacing 5-(piperidin-1-yl)pyridine-2-amino with 5-((4-cyclopropionylpiperazin-1-yl)methyl)pyridine-2-amino, following the same method as compound 20. 1H NMR (300MHz, CDCl3) δ8.61 (s, 1H), 8.54 (d, J = 3.2Hz, 1H), 8.39 (dd, J = 8.5, 0.8Hz, 1H), 8.33-8.29(m,1H),8.07-8.01(m,1H),7.81-7.73(m,3H),4.44(d,J=13.5Hz,1H),3.78 -3.58(m,6H),3.55(s,2H),3.15(t,J=12.0Hz,1H),2.82(s,2H),2.50(s,4H),2.22-2. 11(m,5H),1.76-1.72(m,1H),1.72-1.61(m,2H),1.02-0.96(m,2H),0.81-0.74(m,2H). 13 C NMR (101MHz, CDCl3) δ190.90,171.99,169.01,158.70,155.51,155.48,152.30,15 2.19,150.47,150.38,149.74,148.74,147.96,147.70,140.87,140.82,139.20,1 27.09, 126.68, 121.87, 121.73, 121.68, 119.18, 119.11, 111.60, 78.19, 59.69, 53.15, 52.62, 48.00, 45.35, 42.00, 41.85, 36.89, 34.58, 33.82, 21.46, 10.91, 7.46.

[0154] Biological evaluation experiments were conducted on the compounds of this invention:

[0155] Example 56: CDK6 / cyclin D1 inhibitory activity test

[0156] Different concentrations of the compound were added to 384-well plates (Perkin Elmer, Cat. No. 6007299), replicated, followed by the addition of CDK6 / cyclin D1 kinase (Invitrogen, Cat. No. PV4437) and substrate ULight-4E-BP1 peptide (Perkin Elmer, Cat. No. TRF0128-M), and mixed thoroughly. After incubation at room temperature for 15 minutes, ATP solution was added to initiate the reaction. After reacting at room temperature for 3 hours, EDTA was added to terminate the reaction, and the antibody Eu-phos-4E-BP1 (Perkin Elmer, Cat. No. TRF0216-M) was added to generate a detection signal. After incubation at room temperature for 1 hour, the results were read using an Envision plate reader (BMGPOLARstar Omega). IC50 analysis was performed using GraphPad Prism 8.0 software (San Diego, USA, RRID: SCR_002798). 50 Values ​​and curve fitting.

[0157] Table 1. IC50 values ​​of the compounds in this invention against CDK6 / cyclin D1 kinase inhibitory activity. 50 value

[0158]

[0159]

[0160] Experimental results showed that compounds 14, 23, 33, 36-39 and 42 exhibited strong inhibitory activity against CDK6 at levels below 25 nM.

[0161] Example 57: Cytotoxicity evaluation and cytoprotective effects of active compounds 14, 23, 33, 36-39 and 42

[0162] Cytotoxicity evaluation of the compound: OP9 cells were inoculated at 1×10⁻⁶ cells. 5 The cells were seeded at a density of [number] cells / mL in 96-well plates. After 24 h, the plates were treated with different concentrations of the compound. A blank control group was set up, and three replicates were set up. After incubation at 37°C and 5% CO2 for 48 h, 20 μL of MTT solution (5 mg / mL) was added to each well. After incubation for another 4 h, the supernatant was discarded, and 150 μL of LDMSO was added to each well. The plates were shaken on a shaker for 10 min, and the absorbance was measured at 490 nm. The concentrations of the compound (CC) were calculated using GraphPad Prism 8.0 software (San Diego, USA, RRID: SCR_002798). 50 value.

[0163] Compound-induced cell protection of 5-FU-induced OP9 cells: OP9 cells were inoculated at 1×10⁻⁶ cells per cell line. 5 Inoculated at a density of 1000 cells / mL in 96-well plates, after 24 h, the plates were treated with different concentrations of the compound. A blank control group was set up, and three replicates were set up. After incubation at 37°C and 5% CO2 for 8 h, 20 μL of 5-FU solution (50 μg / mL) was added to each well. After incubation for another 48 h, 20 μL of MTT solution (5 mg / mL) was added to each well. After incubation for another 4 h, the supernatant was discarded, and 150 μL of LDMSO was added to each well. The plates were shaken on a shaker for 10 min, and the absorbance was measured at 490 nm. EC50 was calculated using GraphPad Prism 8.0 software (San Diego, USA, RRID: SCR_002798). 50 value.

[0164] Table 2. Effects of active compounds 14, 23, 33, 36-39, and 42 in this invention on CC in OP9 cells. 50 Values ​​and EC50 values ​​for 5-FU-induced OP9 cell damage 50 value

[0165]

[0166] Experimental results showed that compound 42 exhibited minimal cytotoxicity (CC) compared to the positive control drug Trilaciclib. 50 >100μM) and the strongest protective effect (EC) 50 =4.02μM).

[0167] Example 58: Interaction between active compound 42 and CDK6 protein

[0168] Using Discovery Studio software, molecular docking was performed between active compound 42 and CDK6 protein (PDB:5L2S). The docking results showed that the chromone spirocyclic backbone, pyrimidine, and secondary amine fragments in compound 42 formed hydrogen bonds with CDK6. Figure 2 As can be seen, the chromogenone spirocyclic skeleton, pyrimidine, and secondary amine fragments in the active compound of the present invention are crucial components of compound 42's CDK4 / 6 inhibitory activity; the absence of any one of these components will affect the activity.

[0169] Example 59: Pharmacokinetic determination of compound 42

[0170] Pharmacokinetic characteristics of compound 42 were tested. Ten SD rats were used in this experiment and divided into two groups (n=5 per group). Compound 42 was administered via tail vein injection (2 mg / kg) and gavage (50 mg / kg), respectively. Blood samples (150 μL) were collected via the retro-orbital venous plexus at predetermined time points (0.05 h, 0.083 h, 0.167 h, 0.25 h, 0.5 h, 0.75 h, 1 h, 2 h, 4 h, 6 h, 9 h, 12 h, and 24 h). The concentration of the compound in the plasma samples of SD rats was determined by LC-MS / MS, and pharmacokinetic parameters were calculated using WinNolin software.

[0171] Table 3. Pharmacokinetic characteristics of compound 42

[0172] The experimental results showed that the half-life of the intravenous injection group was 1.54 h, the mean residence time was 1.55 h, and the AUC was 591.10 h·ng / mL; the half-life of the oral administration group was 7.13 h, the Tmax was 6 h, the Cmax was 480.55 ng / mL, the AUC was 4215.57 h·ng / mL, and the oral bioavailability was 28.53%. These results indicate that compound 42 has good drug-like properties.

[0173] Example 60: Pharmacodynamic study of compound 42

[0174] Balb / c mice were randomly divided into 5 groups (n=10): control group, 5-FU group, 5-FU + Trilaciclib group (50 mg / kg, intraperitoneal injection), and 5-FU + 42 group (12.5 mg / kg, 25 mg / kg, gavage). 42 or Trilaciclib was administered one day before inducing bone marrow suppression, followed by a single intraperitoneal injection of 5-FU (100 mg / kg) on ​​day 1. Control mice received an equal volume of physiological saline intraperitoneally or by gavage at the time of administration or model induction. Blood samples were collected from the fundus at designated time points (days 4, 6, 8, and 14) and analyzed using a BC-5000Vet fully automated blood analyzer (Mindray, China). Figure 3 The experimental results showed that, compared with the positive control drug Trilaciclib, the 42 treatment group significantly increased the number of platelets (PLT), red blood cells (RBC), hemoglobin (HGB), white blood cells (WBC), lymphocytes (Lym), and neutrophils (Neu), and improved chemotherapy-induced weight loss and survival rate in mice.

[0175] Example 61: Safety evaluation of compound 42

[0176] Acute toxicity test: Half-sex Balb / c mice were randomly divided into four groups: male control group (n=5), male 42 treatment group (n=10), female control group (n=5), and female 42 treatment group (n=10). After a single oral administration of 42 (1000 mg / kg), mice were weighed every two days. After 14 days, the mice were euthanized using the cervical dislocation method, and the major organs (heart, liver, spleen, lungs, and kidneys) were removed and weighed. The results showed that there was no difference in food intake between the control group and the 42 treatment group. Figure 4 Furthermore, the weights of the mice's major organs, including the heart, liver, spleen, lungs, and kidneys, did not differ between the control and treatment groups. Figure 4 H&E staining showed no obvious lesions in the major organs of mice in the 1000 mg / kg group. Figure 5 The above results indicate that 42 has good drug safety; therefore, the chromogenone spirocyclic compound of the present invention has selective CDK4 / 6 inhibitory activity, which can alleviate and / or treat chemotherapy-induced bone marrow suppression, improve chemotherapy-induced weight loss and improve survival rate, and has great clinical application prospects.

Claims

1. A chromogenone spirocyclic compound, characterized in that, The compound has the structure shown in Formula I: ； Where X is selected from (CH2). n Or C(O), n is 0 or 1; R1 and R2 are selected from C4-C6 cycloalkyl or C6 heterocyclic groups; R3 is a halogen; R4 is a C6 heterocyclic group.

2. The chromogenone spirocyclic compound according to claim 1, characterized in that, X is selected from: C(O) or (CH2). n n is 0 or 1; R1 and R2 are selected from cyclobutyl, cyclopentyl, cyclohexyl, tetrahydropyran, tetrahydrothiothioran or N-amide piperidine; R3 is F; R4 is selected from piperidine, morpholine or piperazine.

3. A chromogenone spirocyclic compound, characterized in that, Selected from the following compounds 1-55: 。 4. The chromogen spirocyclic compound according to claim 1 or claim 3, characterized in that, It also includes pharmaceutically acceptable salts of the aforementioned chromone spirocyclic compounds, specifically salts of compounds of general formula (I) formed by adding an acid, wherein the acid used for salt formation is selected from inorganic or organic acids, wherein the inorganic acid is selected from hydrochloric acid, sulfuric acid, phosphoric acid, and methanesulfonic acid; and the organic acid is selected from galactosic acid, D-glucuronic acid, glycerophosphate, hippuric acid, hydroxyethanesulfonic acid, lactobionic acid, maleic acid, naphthalene-2-sulfonic acid, neopentanoic acid, terephthalic acid, cholic acid, benzenesulfonic acid, citric acid, lactic acid, malic acid, malonic acid, mandelic acid, acetic acid, propionic acid, tartaric acid, succinic acid, formic acid, methanesulfonic acid, picric acid, L-pyroglutamic acid, salicylic acid, gentian acid, p-toluenesulfonic acid, valeric acid, palmitic acid, stearic acid, lauric acid, adipic acid, 4-benzenesulfonic acid, fumaric acid, 3-hydroxynaphthalene-2-carboxylic acid, ascorbic acid, camphoric acid, camphorsulfonic acid, cinnamic acid, dichloroacetic acid, and ethanesulfonic acid.

5. A method for preparing the chromogenone spirocyclic compound according to claim 1 or claim 3, characterized in that, The manufacturing method is as follows: (1) 1-(4-bromo-2-hydroxyphenyl) ethyl ketone shown in Formula II undergoes a condensation reaction with a cyclic ketone under alkaline conditions to give the chromone intermediate compound III; (2) The intermediate compound III of chromone reacts with dipinacolborate in the presence of a palladium catalyst to give compound IV; (3) Compound IV was coupled with a chloropyrimidine derivative to give compound V; (4) Compound V and a 2-aminopyrimidine derivative were coupled together with a palladium catalyst and a phosphine ligand to obtain the chromogenone spirocyclic compound shown in Formula I. The synthetic route is as follows, wherein R1, R2, R3, R4, and X are as described in claim 1: 。 6. The manufacturing method according to claim 5, characterized in that, In step (1), the base used for alkaline conditions is selected from pyrrolidine, piperidine, tetrahydropyridine, sodium hydroxide, potassium hydroxide, potassium carbonate or sodium carbonate; the reaction solvent is selected from methanol, ethanol, isopropanol, perfluoroisopropanol, n-butanol or tert-butanol. Step (2) is carried out under alkaline conditions. The base used is selected from potassium phosphate, potassium carbonate, sodium carbonate, cesium carbonate, potassium fluoride, potassium acetate, sodium acetate, potassium tert-butoxide, or sodium tert-butoxide. The palladium catalyst is selected from Pd(dppf)Cl2, Pd(PPh3)2Cl2, Pd(PPh3)4, Pd2(dba)3, PdCl2(dippf)2, or [Pd(allyl)Cl]2. The reaction solvent is selected from dioxane, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, acetonitrile, or N-methylpyrrolidone. Step (3) is carried out under alkaline conditions, and the base used is selected from sodium hydroxide, potassium hydroxide, barium hydroxide, potassium phosphate, potassium carbonate, sodium carbonate, cesium carbonate, potassium fluoride, potassium acetate, sodium acetate, sodium methoxide, potassium methoxide, sodium ethoxide, potassium ethoxide, potassium tert-butoxide, or sodium tert-butoxide; the palladium catalyst used in the coupling reaction is selected from Pd(dppf)Cl2, Pd(PPh3)2Cl2, Pd(PPh3)4, Pd2(dba)3, PdCl2(dippf)2, or [Pd(allyl)Cl]2; Step (4) is carried out under alkaline conditions. The base used is selected from sodium hydroxide, potassium hydroxide, potassium carbonate, sodium carbonate, cesium carbonate, sodium methoxide, potassium methoxide, sodium ethoxide, potassium ethoxide, potassium tert-butoxide, or sodium tert-butoxide. The palladium catalyst is selected from Pd2(dba)3 or Pd(OAc)2. The phosphine ligand is selected from PCy3, P(t-Bu)3, P(o-tolyl)3, DPPF, JohnPhos, SPhos, XantPhos, or BINAP. The reaction solvent is selected from toluene, xylene, dioxane, tetrahydrofuran, ethylene glycol dimethyl ether, N,N-dimethylformamide, N,N-dimethylacetamide, or dimethyl sulfoxide.

7. The use of the chromogenone spirocyclic compound of claim 1 or claim 3 in the preparation of CDK4 / 6 inhibitors.

8. Use of the chromogenone spirocyclic compound of claim 1 or claim 3 in the preparation of a medicament for relieving and / or treating chemotherapy-induced myelosuppression.

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