PARP/PI3K double-target inhibit containing pyridopyrimidine structure

Abstract

The present disclosure relates to the field of pharmaceutical chemistry, in particular to a class of PARP/PI3K double-target inhibitors (I) containing structures of 5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine and phthalazin-1(2H)-one and a preparation method thereof. As proved by pharmacodynamic tests, the compounds of the present disclosure have PARP/PI3K double-target inhibitory activity and can be used for anti-tumor.

Claims

1. A compound of a formula I or a pharmaceutically acceptable salt thereof: ##STR00020## wherein R.sup.1 represents ##STR00021## R.sup.2 represents ##STR00022##  and R.sup.3 represents H, F, Br, Cl, CF.sub.3, CN, CH.sub.3 or OCH.sub.3; X represents CH or N; Y represents O, NH, NCH.sub.3 or CH.sub.2; and m=1 or 2.

2. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sup.1 represents ##STR00023##

3. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sup.2 represents ##STR00024##

4. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is prepared by the following reaction: ##STR00025## wherein R.sup.1 and R.sup.2 are as defined in claim 1.

5. The compound or the pharmaceutically acceptable salt thereof according to claim 4, wherein the Suzuki reaction is performed under the conditions of adding a catalyst, a base and a reaction solvent, and the catalyst is selected from bis(triphenylphosphine)palladium dichloride, tetrakis(triphenylphosphine)palladium or [1,1′-bis(diphenylphosphino)ferrocene]palladium dichloride; the base is selected from sodium ethoxide, sodium acetate, potassium acetate, potassium phosphate, sodium carbonate or potassium carbonate; the reaction solvent is selected from N,N-dimethylformamide, N,N-dimethylacetamide, ethylene glycol dimethyl ether, dioxane, tetrahydrofuran, toluene, ethanol, water or a mixed solvent of any two or three of the solvents; and a reaction temperature is 80° C. to 120° C.

6. The compound or the pharmaceutically acceptable salt thereof according to claim 5, wherein the catalyst is the tetrakis(triphenylphosphine)palladium; the base is the potassium carbonate; the solvent is a mixed solvent of the dioxane and the water; and the reaction temperature is 100° C. to 110° C.

7. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein the pharmaceutically acceptable salt is an acid addition salt of the compound of the formula (I), wherein an acid used for forming the salt is: hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or ferulic acid.

8. A pharmaceutical composition, comprising the compound or the pharmaceutically acceptable salt thereof according to claim 1 and a pharmaceutically acceptable carrier.

9. A pharmaceutical drug comprising a therapeutically effective amount of the compound or the pharmaceutically acceptable salt thereof according to claim 1 for treating a tumor, wherein the pharmaceutical drug is capable of inhibiting activities of PARP-1 and PI3K in the tumor.

10. The pharmaceutical drug according to claim 9, wherein the pharmaceutical drug is used for treating a subject with the tumor that expresses the activities of the PARP-1 or/and PI3K.

11. A method for treating a cancer comprising a step of administrating a therapeutically effective amount of the compound or the pharmaceutically acceptable salt thereof of claim 1 to a subject in need, wherein cell of the cancer over-expresses the PARP-1 or/and PI3K.

12. The method according to claim 11, wherein the cancer is selected from the group consisting of a colon cancer, a breast cancer and an ovarian cancer.

Description

DETAILED DESCRIPTION OF THE INVENTION

Embodiment 1

Synthesis of 4-(3-(2-(2-aminopyrimidin-5-yl)-4-morpholinyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (I-1)

7-benzyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-2,4(1H,3H)-dione (2)

(1) Ethyl 1-benzyl-3-oxopiperidine-4-carboxylate hydrochloride (1) (40.0 g, 134.33 mmol) and urea (17.09 g, 282.01 mmol) were added into a 500 mL three-necked flask, 250 mL of absolute methanol was added and stirred to be dissolved, the temperature was reduced to 0° C., and a methanol solution of sodium methoxide (228.0 mL, 1.0 mol/L) was dropwise added. After dropwise addition, heating reflux reaction under nitrogen protection was performed for about 20 hours. TLC (petroleum ether: ethyl acetate=5:1) was adopted to detect that the raw material 1 was completely reacted, heating was stopped, a mixture was cooled to the room temperature, a large amount of white solids were precipitated, and the temperature was reduced to 0° C. Stirring was continued for 1 hour, suction filtration was carried out, and a filter cake was washed with 50 mL of methanol to obtain a white solid. The white solid was dissolved in 400 mL of water and adjusted to be neutral with 1 mol/L hydrochloric acid, and a large amount of solids were precipitated. Suction filtration and drying were carried out to obtain 27.68 g of a white solid with the yield of 80.1%. Without purification, the white solid was directly applied to the next step.

7-benzyl-2,4-dichloro-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine (3)

(2) 200 mL of phosphorus oxychloride was added to a 500 mL three-necked flask, and the temperature was reduced to 0° C. A compound 2 (27.68 g, 107.58 mmol) was slowly added. After the addition, a reaction solution was pink and cloudy. Under nitrogen protection, heating reflux reaction was performed for 6-8 hours. TLC (petroleum ether: ethyl acetate=3:1) was adopted to detect that the raw material 2 was completely reacted, heating was stopped, a mixture was slightly cooled, and the phosphorus oxychloride was evaporated under reduced pressure. A residue was slowly poured into 400 g of crushed ice, the pH was adjusted to 8 to 9 with a 5 mol/L NaOH solution, and a solid was precipitated. Suction filtration was carried out, and a filter cake was washed with water and dried to obtain 29.0 g of an off-white solid with the yield of 91.6%. .sup.1HNMR (300 MHz, CDCl.sub.3) δ (ppm): 7.34-7.29 (5H, m, ArH), 3.75 (2H, s, CH.sub.2), 3.67 (2H, s, CH.sub.2), 2.85 (4H, m, 2×CH.sub.2).

4-(7-benzyl-2-chloro-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)morpholine (II-1)

(3) The compound 3 (29.0 g, 98.58 mmol) was dissolved in a mixed solvent of isopropanol (200 mL) and dichloromethane (20 mL), and morpholine (10.31 g, 118.30 mmol) and DIEA (32.59 mL, 197.16 mmol) were slowly added. After the addition, the temperature was raised to 50° C. for reaction for 3 to 5 hours, TLC (petroleum ether: ethyl acetate=9:1) was adopted to detect that the raw material 3 was completely reacted, heating was stopped, the temperature was reduced to the room temperature, a solid was precipitated, and the temperature was reduced to 0° C. Stirring was continued for 30 minutes, suction filtration was carried out, and a filter cake was washed with 50 mL of isopropanol and dried to obtain 23.0 g of an off-white solid. A filtrate was concentrated under reduced pressure, a residue was dissolved with 200 mL of ethyl acetate and washed with water (100 mL) and a saturated sodium chloride solution (100 mL) in sequence, and concentration under reduced pressure was performed to obtain a yellow grease. Column chromatography purification (petroleum ether: ethyl acetate=50:1 to 5:1) was performed to obtain 7.42 g of a white solid, and 30.42 g of a product was obtained in total with the yield of 89.5%. m.p. 154-156° C. .sup.1HNMR (300 MHz, DMSO-d.sub.6) δ (ppm): 7.34-7.23 (5H, m, ArH), 3.66-3.62 (6H, m, 2×CH.sub.2), 3.47-3.44 (6H, m, 3×CH.sub.2), 2.65-2.54 (4H, m, 2×CH.sub.2).

4-(2-chloro-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)morpholine (III-1)

(4) A compound 4 (21.10 g, 61.19 mmol) was dissolved in 200 mL of dichloromethane and cooled to 0° C. in an ice bath, 1-chloroethyl chloroformate (26.41 mL, 244.76 mmol) was added dropwise, and after the addition, a mixture was stirred with heat preservation for 30 minutes. The temperature was raised to 25° C., and stirring was performed for reaction for 8 to 10 hours. TLC (petroleum ether: ethyl acetate=1:1) was adopted to detect that the raw material 4 was completely reacted, and the solvent was evaporated under reduced pressure. 200 mL of methanol was added for reflux reaction for 1 hour, and the solvent was evaporated under reduced pressure. 200 mL of water was added to dissolve a residue, and the pH was adjusted to 8 to 9 with a 1 mol/L NaOH solution. Dichloromethane (150 mL×3) was adopted for extraction, an organic layer was combined, a saturated sodium chloride solution (200 mL×2) was adopted for washing, and drying was performed with anhydrous Na.sub.2SO.sub.4. Suction filtration was carried out, a filtrate was concentrated, and a residue was subjected to column chromatography purification (dichloromethane:methanol=80:1 to 20:1 for gradient elution) to obtain 10.5 g of a yellow solid with the yield of 67.4%. .sup.1HNMR (300 MHz, DMSO-d.sub.6) δ (ppm): 3.75-3.64 (6H, m, 3×CH.sub.2), 3.44-3.41 (4H, m, 2×CH.sub.2), 2.79 (2H, m, CH.sub.2), 2.66 (1H, s, NH), 2.50-2.48 (2H, m, CH.sub.2).

4-(3-(2-chloro-4-morpholinyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (IV-1)

(5) 2-fluoro-5-((4-oxo-3,4-dihydrophthalazin)methyl)benzoic acid (4) (11.24 g, 37.68 mmol), 4-(2-chloro-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)morpholine (III-1) (9.60 g, 37.69 mmol) and PyBOP (23.54 g, 45.23 mmol) were added into a 200 mL three-necked flask. 100 mL of DMF was added and stirred to be dissolved, and then DIEA (24.92 mL, 150.79 mmol) was added and stirred at 25° C. for reaction for 6-8 hours. TLC (dichloromethane:methanol=20:1) was adopted to detect that the raw material 4 was completely reacted, a reaction solution was poured into 300 mL of water, and a large amount of solids were precipitated. Suction filtration was carried out, a filter cake was washed with 100 mL of water and dried to obtain a crude product, and column chromatography purification (dichloromethane:methanol=100:1 to 20:1 for gradient elution) was performed to obtain 12.06 g of an off-white solid with the yield of 59.8%. m.p. 174-177° C. .sup.1HNMR (300 MHz, DMSO-d.sub.6) δ (ppm): 12.61 (1H, s, CONH), 8.29-8.25 (1H, m, ArH), 7.98 (1H, d, J=7.7 Hz, ArH), 7.92-7.80 (2H, m, ArH), 7.52-7.40 (2H, m, ArH), 7.32-7.24 (1H, m, ArH), 4.63 (1H, s, 0.5×CH.sub.2), 4.36 (2H, s, CH.sub.2), 4.29 (1H, s, 0.5×CH.sub.2), 3.85-3.61 (5H, m, 2.5×CH.sub.2), 3.54-3.42 (4H, m, 2×CH.sub.2), 3.29-3.13 (1H, m, 0.5×CH.sub.2), 2.74-2.57 (2H, m, CH.sub.2).

4-(3-(2-(2-aminopyrimidin-5-yl)-4-morpholinyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (I-1)

(6) The compound IV-1 (9.06 g, 16.94 mmol) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (V-1) (5.61 g, 25.40 mmol) were added into a 250 mL reaction flask, and 100 mL of dioxane was added and stirred to be dissolved. K.sub.2CO.sub.3 (9.37 g, 67.76 mmol) was dissolved in 10 mL of water, a reaction solution was added slowly, and then tetratriphenylphosphine palladium (0.98 g, 0.85 mmol) was added. Under nitrogen protection, heating reflux reaction was performed for 4 to 6 hours to precipitate a yellow solid. TLC (dichloromethane:methanol=20:1) was adopted to detect that the raw material IV-1 was completely reacted, and a mixture was cooled to the room temperature. Suction filtration was performed, and a filter cake was washed with water (40 mL) and ethyl acetate (20 mL) in sequence and dried to obtain a crude product. 120 mL of ethyl acetate was added and beaten for 2 hours, and suction filtration and drying were carried out to obtain 5.80 g of a light-yellow solid with the yield of 57.7%. m.p. 159-160° C. .sup.1HNMR (300 MHz, DMSO-d.sub.6) δ (ppm): 12.73 (1H, s, CONH), 9.16 (1H, s, ArH), 9.07 (1H, s, ArH), 8.37 (1H, t, J=6.8 Hz, ArH), 8.11-7.88 (3H, m, ArH), 7.63-7.54 (2H, m, ArH), 7.40 (1H, t, J=8.7 Hz, ArH), 7.30 (2H, s, NH.sub.2), 4.81 (1H, s, 0.5×CH.sub.2), 4.47 (2H, s, CH.sub.2), 4.45 (1H, s, 0.5×CH.sub.2), 3.95-3.55 (10H, m, 5×CH.sub.2), 2.85-2.71 (2H, m, CH.sub.2). .sup.13CNMR (75 MHz, DMSO-d.sub.6) δ (ppm): 164.12, 163.67, 162.66, 160.27, 159.89, 159.34, 157.75, 154.92, 144.85, 134.85, 133.44, 132.01, 131.54, 129.05, 127.85, 126.03, 125.43, 123.67, 119.47, 117.77, 115.88, 112.42, 65.97, 50.45, 47.69, 46.43, 36.44, 26.14. HRMS (ESI): m/z [M+H].sup.+. Calcd for C.sub.31H.sub.28FN.sub.9O.sub.3: 594.2372; Found: 594.2371.

Embodiment 2

Synthesis of 4-(3-(2-(6-aminopyrimidin-3-yl)-4-morpholinyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (I-2)

(7) A compound IV-1 (300 mg, 0.56 mmol) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (V-2) (136 mg, 0.62 mmol) were added into a 100 mL three-necked flask, and 20 mL of dioxane was added and stirred to be dissolved. K.sub.2CO.sub.3 (310 mg, 2.24 mmol) was dissolved in 2 mL of water, a reaction solution was added, and then tetratriphenylphosphine palladium (65 mg, 0.06 mmol) was added. Under nitrogen protection, heating reflux reaction was performed for 4 to 5 hours. TLC (dichloromethane:methanol=20:1) was adopted to detect that the raw material IV-1 was completely reacted, heating was stopped, and a mixture was cooled to the room temperature. Suction filtration was performed, a filtrate was concentrated, a residue was added into 40 mL of ethyl acetate, a mixed solution was washed with water (20 mL×1) and a saturated sodium chloride solution (20 mL×2) in sequence, and drying was performed with anhydrous Na.sub.2SO.sub.4. Suction filtration was performed, a filtrate was concentrated, and a residue was subjected to column chromatography purification (dichloromethane:methanol=100:1 to 30:1) to obtain 220 mg of a light-yellow solid with the yield of 66.3%. m.p. 206-207° C. .sup.1HNMR (300 MHz, DMSO-d.sub.6) δ (ppm): 12.61 (1H, s, CONH), 8.89-8.80 (1H, m, ArH), 8.28-8.12 (2H, m, ArH), 8.00-7.78 (3H, m, ArH), 7.51-7.45 (2H, m, ArH), 7.29 (1H, t, J=9.1 Hz, ArH), 6.51-6.47 (1H, m, ArH), 6.44 (2H, s, NH.sub.2), 4.69 (1H, s, 0.5×CH.sub.2), 4.36 (2H, s, CH.sub.2), 4.33 (1H, s, 0.5×CH.sub.2), 3.88-3.47 (10H, m, 5×CH.sub.2), 2.73-2.58 (2H, m, CH.sub.2). .sup.13CNMR (75 MHz, DMSO-d.sub.6) δ (ppm): 164.39, 163.78, 161.00, 160.22, 159.84, 159.37, 154.97, 148.57, 144.84, 136.17, 134.88, 133.43, 132.04, 131.52, 129.07, 127.88, 126.04, 125.42, 123.77, 121.29, 116.10, 115.83, 111.90, 107.18, 65.98, 47.77, 46.51, 43.74, 36.47, 26.03. HRMS (ESI): m/z [M+H].sup.+. Calcd for C.sub.32H.sub.29FN.sub.8O.sub.3: 593.2419; Found: 593.2418.

Embodiment 3

Synthesis of 4-(3-(2-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-4-morpholinyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (1-3)

(8) With the compound IV-1 (1.0 g, 1.87 mmol) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4-(trifluoromethyl)pyridin-2-amine (V-3) (646 mg, 2.24 mmol) as raw materials, operation was the same as the method of I-2, and column chromatography purification (dichloromethane:methanol=100:1 to 20:1 for gradient elution) was carried out to obtain 680 mg of a light-yellow solid with the yield of 55.1%. m.p. 158-160° C. .sup.1HNMR (300 MHz, DMSO-d.sub.6) δ (ppm): 12.56 (1H, s, CONH), 8.51-8.42 (1H, m, ArH), 8.25 (1H, t, J=7.3 Hz, ArH), 7.98-7.75 (3H, m, ArH), 7.50-7.43 (2H, m, ArH), 7.30-7.22 (1H, m, ArH), 6.82 (1H, s, ArH), 6.80 (2H, s, NH.sub.2), 4.68 (1H, s, ½ArCH.sub.2N), 4.35 (2H, s, ArCH.sub.2), 4.32 (1H, s, ½ArCH.sub.2N), 3.84-3.39 (10H, m, 5CH.sub.2), 2.76-2.60 (2H, m, NCH.sub.2CH.sub.2). .sup.13CNMR (75 MHz, DMSO-d.sub.6) δ (ppm): 164.43, 164.03, 163.55, 160.50, 159.82, 159.37, 158.22, 154.98, 152.35, 144.79, 135.56, 134.89, 133.38, 131.96, 131.47, 129.08, 127.90, 126.03, 125.38, 123.47, 121.34, 119.67, 116.10, 112.56, 104.37, 66.00, 47.70, 46.29, 43.64, 36.50, 25.99. HRMS (ESI): m/z [M+H].sup.+. Calcd for C.sub.33H.sub.28F.sub.4N.sub.8O.sub.3: 661.2293; Found: 661.2298.

Embodiment 4

Synthesis of 4-(3-(2-(6-amino-5-(trifluoromethyl)pyridin-3-yl)-4-morpholinyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (I-5)

(9) With the compound IV-1 (300 mg, 0.56 mmol) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-(trifluoromethyl)pyridin-2-amine (V-4) (178 mg, 0.62 mmol) as raw materials, operation was the same as the method of 1-2, and column chromatography purification (dichloromethane:methanol=100:1 to 40:1) was carried out to obtain 250 mg of a light-yellow solid with the yield of 67.6%. m.p. 256-258° C. .sup.1HNMR (300 MHz, DMSO-d.sub.6) δ (ppm): 12.61 (1H, s, CONH), 9.11-9.03 (1H, m, ArH), 8.51-8.41 (1H, m, ArH), 8.26 (1H, t, J=7.2 Hz, ArH), 8.00-7.76 (3H, m, ArH), 7.56-7.44 (2H, m, ArH), 7.29 (1H, t, J=8.7 Hz, ArH), 7.01 (2H, s, NH.sub.2), 4.71 (2H, s, CH.sub.2), 4.36 (2H, s, CH.sub.2), 3.93-3.39 (10H, m, 5×CH.sub.2), 2.75-2.61 (2H, m, CH.sub.2). .sup.13CNMR (75 MHz, DMSO-d.sub.6) δ (ppm): 164.40, 163.93, 160.42, 160.06, 159.35, 157.93, 156.51, 152.23, 144.81, 137.91, 134.97, 133.87, 133.41, 131.94, 131.50, 129.61, 129.09, 128.84, 127.90, 126.04, 125.43, 123.71, 120.89, 116.11, 112.64, 65.96, 47.72, 46.48, 43.71, 36.47, 26.12. HRMS (ESI): m/z [M+H].sup.+. Calcd for C.sub.33H.sub.28F.sub.4N.sub.8O.sub.3: 661.2293; Found: 661.2290.

Embodiment 5

Synthesis of 4-(3-(2-(4-amino-2-fluorophenyl)-4-morpholinyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (I-6)

(10) With the compound IV-1 (300 mg, 0.56 mmol) and 3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (V-5) (147 mg, 0.62 mmol) as raw materials, operation was the same as the method of I-2, and column chromatography purification (dichloromethane:methanol=100:1 to 50:1 for gradient elution) was carried out to obtain 145 mg of a light-yellow solid with the yield of 42.5%. m.p. 204-206° C. .sup.1HNMR (300 MHz, DMSO-d.sub.6) δ (ppm): 12.58 (1H, s, CONH), 8.28-8.22 (1H, s, ArH), 8.00-7.68 (4H, m, ArH), 7.51-7.43 (2H, m, ArH), 7.30-7.25 (1H, m, ArH), 6.44-6.27 (2H, m, ArH), 5.81 (2H, s, NH.sub.2), 4.65 (1H, s, 0.5×CH.sub.2), 4.35 (2H, s, CH.sub.2), 4.30 (1H, s, 0.5×CH.sub.2), 3.82-3.38 (10H, m, 5×CH.sub.2), 2.72-2.57 (2H, m, CH.sub.2). .sup.13CNMR (75 MHz, DMSO-d.sub.6) δ (ppm): 164.37, 163.99, 163.53, 160.07, 159.36, 152.55, 144.84, 134.88, 133.43, 132.10, 131.53, 129.06, 128.79, 127.87, 126.04, 125.43, 123.77, 123.50, 121.31, 115.85, 112.61, 111.47, 109.46, 100.48, 100.09, 65.96, 47.76, 46.44, 43.70, 36.46, 25.96. HRMS

(11) (ESI): m/z [M+H].sup.+. Calcd for C.sub.33H.sub.29F.sub.2N.sub.7O.sub.3: 610.2373; Found: 610.2368.

Embodiment 6

Synthesis of 4-(3-(2-(3-amino-4-fluorophenyl)-4-morpholinyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (I-7)

(12) With the compound IV-1 (300 mg, 0.56 mmol) and 2-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (V-6) (147 mg, 0.62 mmol) as raw materials, operation was the same as the method of I-2, and column chromatography purification (dichloromethane:methanol=100:1 to 50:1 for gradient elution) was carried out to obtain 160 mg of a light-yellow solid with the yield of 46.9%. m.p. 236-239° C. .sup.1HNMR (300 MHz, DMSO-d.sub.6) δ (ppm): 12.59 (1H, s, CONH), 8.25 (1H, t, J=6.6 Hz, ArH), 7.97 (1H, d, J=7.8 Hz, ArH), 7.91-7.71 (3H, m, ArH), 7.58-7.42 (3H, m, ArH), 7.29 (1H, t, J=9.0 Hz, ArH), 7.15-7.04 (1H, m, ArH), 4.74 (1H, s, 0.5×CH.sub.2), 4.38 (1H, s, 0.5×CH.sub.2), 4.35 (2H, m, CH.sub.2), 3.84-3.39 (10H, m, 5×CH.sub.2), 2.77-2.62 (2H, m, CH.sub.2). .sup.13CNMR (75 MHz, DMSO-d.sub.6) δ (ppm): 164.43, 164.02, 163.28, 159.36, 158.34, 154.99, 144.80, 138.50, 134.89, 133.42, 132.18, 131.52, 129.09, 128.81, 127.90, 126.04, 125.42, 123.55, 117.40, 116.62, 116.15, 115.87, 115.18, 114.93, 112.34, 66.01, 47.81, 45.59, 43.55, 36.48, 26.21. HRMS (ESI): m/z [M+H].sup.+. Calcd for C.sub.33H.sub.29F.sub.2N.sub.7O.sub.3: 610.2373; Found: 610.2374.

Embodiment 7

Synthesis of 4-(3-(2-(1H-indazole-4-yl)-4-morpholinyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (I-8)

(13) With the compound IV-1 (300 mg, 0.56 mmol) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (V-7) (151 mg, 0.62 mmol) as raw materials, operation was the same as the method of 1-2, and column chromatography purification (dichloromethane:methanol=100:1 to 40:1) was carried out to obtain 130 mg of a light-yellow solid with the yield of 37.6%. m.p. 198-200° C. .sup.1HNMR (300 MHz, DMSO-d.sub.6) δ (ppm): 13.24 (1H, s, NH), 12.60 (1H, s, CONH), 8.82-8.70 (1H, m, ArH), 8.28-8.07 (2H, m, ArH), 7.97 (t, J=7.2 Hz, 1H), 7.91-7.72 (2H, m, ArH), 7.67 (1H, t, J=7.0 Hz, ArH), 7.50-7.40 (3H, m, ArH), 7.32-7.25 (1H, m, ArH), 4.83 (1H, s, 0.5×CH.sub.2), 4.46 (1H, s, 0.5×CH.sub.2), 4.36 (2H, s, CH.sub.2), 3.88-3.49 (10H, m, 5×CH.sub.2), 2.80-2.66 (2H, m, CH.sub.2). .sup.13CNMR (75 MHz, DMSO-d.sub.6) δ (ppm): 169.67, 164.47, 164.19, 160.48, 159.36, 158.23, 154.85, 144.82, 140.74, 134.99, 133.42, 131.93, 131.52, 130.29, 129.10, 127.91, 126.05, 125.42, 123.76, 123.51, 121.14, 120.79, 116.12, 115.83, 113.43, 112.42, 66.03, 48.04, 46.56, 43.68, 36.49, 26.18. HRMS (ESI): m/z [M+H].sup.+. Calcd for C.sub.34H.sub.29FN.sub.8O.sub.3: 617.2419; Found: 617.2425.

Embodiment 8

Synthesis of 4-(3-(2-(1H-indole-5-yl)-4-morpholinyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (I-9)

(14) With the compound IV-1 (300 mg, 0.56 mmol) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole (V-8) (151 mg, 0.62 mmol) as raw materials, operation was the same as the method of 1-2, and column chromatography purification (dichloromethane:methanol=100:1 to 60:1 for gradient elution) was carried out to obtain 153 mg of a light-yellow solid with the yield of 44.4%. m.p. 238-240° C. .sup.1HNMR (300 MHz, DMSO-d.sub.6) δ (ppm): 12.59 (1H, s, CONH), 11.24 (1H, s, NH), 8.60-8.51 (1H, m, ArH), 8.26 (1H, t, J=6.5 Hz, ArH), 8.18-8.05 (1H, m, ArH), 8.00-7.75 (3H, m, ArH), 7.50-7.36 (4H, m, ArH), 7.28 (1H, t, J=8.9 Hz, ArH), 6.53 (1H, d, J=9.6 Hz, ArH), 4.73 (1H, s, 0.5×CH.sub.2), 4.38 (1H, s, 0.5×CH.sub.2), 4.36 (2H, s, CH.sub.2), 3.86-3.40 (10H, m, 5×CH.sub.2), 2.75-2.61 (2H, m, CH.sub.2). .sup.13CNMR (75 MHz, DMSO-d.sub.6) (ppm): 164.41, 164.05, 161.34, 160.29, 159.92, 159.36, 151.00, 144.83, 137.37, 134.99, 133.43, 131.90, 131.52, 129.09, 128.49, 127.90, 127.59, 126.05, 125.44, 121.11, 120.19, 117.44, 116.13, 112.10, 111.01, 102.17, 90.23, 66.02, 47.87, 46.56, 43.76, 36.50, 26.05. HRMS (ESI): m/z [M+H].sup.+. Calcd for C.sub.35H.sub.30FN.sub.7O.sub.3: 616.2467; Found: 616.2462.

Embodiment 9

Synthesis of 4-(3-(2-(1H-indole-4-yl)-4-morpholinyl-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (I-10)

(15) With the compound IV-1 (300 mg, 0.56 mmol) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indole (V-9) (151 mg, 0.62 mmol) as raw materials, operation was the same as the method of 1-2, and column chromatography purification (dichloromethane:methanol=100:1 to 60:1 for gradient elution) was carried out to obtain 125 mg of an off-white solid with the yield of 36.3%. m.p. 178-181° C. .sup.1HNMR (300 MHz, DMSO-d.sub.6) δ (ppm): 12.62 (1H, s, CONH), 11.29 (1H, s, NH), 8.26 (1H, t, J=8.9 Hz, ArH), 8.12 (1H, d, J=6.9 Hz, ArH), 8.02-7.74 (4H, m, ArH), 7.55-7.40 (4H, m, ArH), 7.34-7.14 (2H, m, ArH), 4.80 (1H, s, 0.5×CH.sub.2), 4.45 (1H, s, 0.5×CH.sub.2), 4.37 (2H, s, CH.sub.2), 3.90-3.46 (10H, m, 5×CH.sub.2), 2.80-2.66 (2H, m, CH.sub.2). .sup.13CNMR (75 MHz, DMSO-d.sub.6) δ (ppm): 164.47, 164.10, 162.16, 160.20, 159.89, 159.35, 158.23, 151.45, 144.82, 140.70, 137.03, 134.91, 133.42, 132.03, 131.51, 129.10, 128.85, 127.89, 126.10, 125.41, 123.79, 120.36, 116.11, 115.85, 113.69, 112.56, 103.32, 66.05, 48.09, 46.59, 43.77, 36.49, 26.05. HRMS (ESI): m/z [M+H].sup.+. Calcd for C.sub.35H.sub.30FN.sub.7O.sub.3: 616.2467; Found: 616.2464.

Embodiment 10

Synthesis of (S)-4-(3-(2-(2-aminopyrimidin-5-yl)-4-(3-methylmorpholinyl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (I-11)

(S)-4-(7-benzyl-2-chloro-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-3-methylmorpholine (II-2)

(16) An intermediate 3 (2.0 g, 6.80 mmol) was dissolved in a mixed solvent of isopropanol (20 mL) and dichloromethane (4 mL), and (S)-3-methylmorpholine (0.83 g, 8.20 mmol) and DIEA (2.24 mL, 13.56 mmol) were slowly added. After the addition, the temperature was raised to 50° C. for reaction for 8 to 10 hours. TLC (petroleum ether: ethyl acetate=9:1) was adopted to detect that a raw material 3 was completely reacted, heating was stopped, and a mixture was slightly cooled. The solvent was evaporated under reduced pressure, a residue was dissolved by 50 mL of ethyl acetate and washed with water (50 mL) and a saturated sodium chloride solution (50 mL×2) in sequence, and drying was performed with anhydrous Na.sub.2SO.sub.4. Suction filtration was carried out, a filtrate was concentrated under reduced pressure to obtain a yellow grease, and column chromatography purification (petroleum ether: ethyl acetate=20:1 to 5:1 for gradient elution) was performed to obtain 1.4 g of a yellow solid with the yield of 57.4%. .sup.1HNMR (300 MHz, CDCl.sub.3) δ (ppm): 7.39-7.30 (5H, m, ArH), 4.18-4.11 (1H, m, 0.5×CH.sub.2), 3.97-3.92 (1H, m, 0.5×CH.sub.2), 3.78-3.44 (9H, m, 4×CH.sub.2, CH), 2.82-2.57 (4H, m, 2×CH.sub.2), 1.34 (3H, d, J=6.8 Hz, CH.sub.3).

(S)-4-(2-chloro-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-3-methylmorpholine (III-2)

(17) An intermediate 11-2 (1.10 g, 3.07 mmol) was dissolved in 20 mL of dichloromethane, and the temperature was reduced to 0° C. 1-chloroethyl chloroformate (1.43 mL, 13.28 mmol) was added dropwise and stirred with heat preservation for 15 min. The temperature was raised to 25° C., and stirring was performed for reaction for 6 to 10 hours. TLC (petroleum ether: ethyl acetate=1:1) was adopted to detect that the raw material II-2 was completely reacted, and a solvent was evaporated under reduced pressure. 20 mL of methanol was added for reflux reaction for 30 minutes. A solvent was evaporated under reduced pressure, a residue was dissolved in 20 mL of water, the pH was adjusted to 8 to 9 with a 1 mol/L NaOH solution, dichloromethane (30 mL×3) was adopted for extraction, an organic layer was combined, a saturated sodium chloride solution (40 mL×3) was adopted for washing, and drying was performed with anhydrous Na.sub.2SO.sub.4. Suction filtration was carried out, a filtrate was concentrated to obtain a brown grease, and column chromatography purification (dichloromethane:methanol=40:1 to 5:1 for gradient elution) was carried out to obtain 550 mg of a yellow solid with the yield of 66.7%. .sup.1HNMR (300 MHz, CDCl.sub.3) δ (ppm): 4.12 (1H, m, 0.5×CH.sub.2), 3.99 (2H, s, CH.sub.2), 3.95-3.91 (1H, m, 0.5×CH.sub.2), 3.78-3.59 (4H, m, 2×CH.sub.2), 3.53-3.43 (1H, m, CH), 3.14-3.07 (1H, m, 0.5×CH.sub.2), 3.00-2.92 (1H, m, 0.5×CH.sub.2), 2.66-2.53 (2H, m, CH.sub.2), 2.29 (1H, s, NH), 1.32 (3H, d, J=6.8 Hz, CH.sub.3).

(S)-4-(3-(2-chloro-4-(3-methylmorpholinyl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (IV-2)

(18) A compound 4 (555 mg, 1.86 mmol), a compound 111-2 (500 mg, 1.86 mmol) and PyBOP (1.16 g, 2.23 mmol) were added into a 100 mL three-necked flask, 20 mL of DMF was added and stirred to be dissolved, and then DIEA (1.08 mL, 6.51 mmol) was added and stirred at 25° C. for reaction for 6-8 hours. TLC (dichloromethane:methanol=20:1) was adopted to detect that the raw material 111-2 was completely reacted, a reaction solution was poured into 60 mL of water, and a yellow solid was precipitated. Suction filtration was carried out, and a filter cake was washed with 20 mL of water and dried to obtain a crude product. Column chromatography purification (dichloromethane:methanol=100:1 to 20:1 for gradient elution) was carried out to obtain 740 mg of a yellow solid with the yield of 72.5%. m.p. 144-147° C. .sup.1HNMR (300 MHz, CDCl.sub.3) δ (ppm): 10.82 (1H, s, CONH), 8.51-8.46 (1H, m, ArH), 7.83-7.70 (3H, m, ArH), 7.45-7.36 (2H, m, ArH), 7.12-7.04 (1H, m, ArH), 4.46 (1H, s, 0.5×CH.sub.2), 4.33 (1H, s, 0.5×CH.sub.2), 4.30 (2H, s, CH.sub.2), 4.17-4.06 (2H, m, CH.sub.2), 3.95 (2H, m, CH.sub.2), 3.73-3.51 (7H, m, 3×CH.sub.2, CH), 1.38 (3H, d, J=6.7 Hz, CH.sub.3).

(S)-4-(3-(2-(2-aminopyrimidin-5-yl)-4-(3-methylmorpholinyl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (I-11)

(19) With the compound IV-2 (300 mg, 0.55 mmol) and the compound V-1 (181 mg, 0.82 mmol) as raw materials, operation was the same as the method of 1-2, and column chromatography purification (dichloromethane:methanol=60:1 to 20:1 for gradient elution) was carried out to obtain 170 mg of a light-yellow solid with the yield of 50.9%. m.p. 214-216° C. .sup.1HNMR (300 MHz, DMSO-d.sub.6) δ (ppm): 12.60 (1H, s, CONH), 9.03 (1H, s, ArH), 8.94 (1H, s, ArH), 8.25 (1H, t, J=6.7 Hz, ArH), 7.99-7.76 (3H, m, ArH), 7.49-7.42 (2H, s, ArH), 7.28 (1H, t, J=9.1 Hz, ArH), 7.15 (2H, s, NH.sub.2), 4.78-4.58 (H, m, 0.5×CH.sub.2), 4.35 (2H, s, CH.sub.2), 4.16-4.04 (H, m, 0.5×CH.sub.2), 3.87-3.38 (9H, m, 4×CH.sub.2, CH), 2.71-2.58 (2H, m, CH.sub.2), 1.23 (3H, d, J=7.2 Hz, CH.sub.3). .sup.13CNMR (75 MHz, DMSO-d.sub.6) δ (ppm): 164.36, 164.13, 163.63, 160.35, 159.97, 159.35, 157.67, 144.83, 134.88, 133.43, 132.01, 131.53, 129.04, 127.87, 126.82, 126.03, 125.44, 123.71, 123.45, 119.53, 116.10, 112.56, 70.27, 66.25, 49.54, 46.52, 43.71, 42.06, 36.45, 26.15, 14.09. HRMS (ESI): m/z [M+H].sup.+. Calcd for C.sub.32H.sub.30FN.sub.9O.sub.3: 608.2528; Found: 608.2535.

Embodiment 11

Synthesis of 4-(3-(2-(2-aminopyrimidin-5-yl)-4-((2S,6R)-2,6-dimethylmorpholinyl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (I-12)

(2S,6R)-4-(7-benzyl-2-chloro-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-2,6-dimethylmorpholine (II-3)

(20) A compound 3 (2.0 g, 6.80 mmol) was dissolved in 30 mL of isopropanol, 4 mL of dichloromethane was added to assist dissolution, and (2R,6S)-2,6-dimethylmorpholine (0.94 g, 8.16 mmol) and DIEA (2.24 mL, 13.56 mmol) were slowly added. After the addition, the temperature was raised to 50° C. for reaction for 8 to 10 hours. TLC (petroleum ether: ethyl acetate=9:1) was adopted to detect that the raw material 3 was completely reacted, and a mixture was slightly cooled. A solvent was evaporated under reduced pressure, a residue was dissolved in 100 mL of ethyl acetate and washed with water (50 mL) and a saturated sodium chloride solution (50 mL×2) in sequence, and drying was performed with anhydrous Na.sub.2SO.sub.4. Suction filtration was carried out, a filtrate was concentrated to obtain 1.86 g of a yellow grease with the yield of 73.2%, and the yellow grease was directly used in the next step of reaction without purification.

(2S,6R)-4-(2-chloro-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-4-yl)-2,6-dimethylmorpholine (III-3)

(21) The compound 11-3 (1.66 g, 4.45 mmol) was dissolved in 20 mL of dichloromethane, and the temperature was reduced to 0° C. 1-chloroethyl chloroformate (2.08 mL, 19.24 mmol) was added dropwise and stirred with heat preservation for 15 minutes. The temperature was raised to 25° C., and stirring was carried out for reaction for 8 to 10 hours. TLC (petroleum ether: ethyl acetate=1:1) was adopted to detect that the raw material II-3 was completely reacted, and a solvent was evaporated under reduced pressure. 20 mL of methanol was added, and heating reflux reaction was performed for 30 minutes. A solvent was evaporated under reduced pressure, a residue was dissolved in 20 mL of water, and the pH was adjusted to 8 to 9 with a 1 mol/L NaOH solution. Dichloromethane (20 mL×3) was adopted for extraction, an organic layer was combined, washing was performed with a saturated sodium chloride solution (30 mL×2), and drying was performed with anhydrous Na.sub.2SO.sub.4. Suction filtration was carried out, a filtrate was concentrated to obtain a brown grease, and column chromatography purification (dichloromethane:methanol=40:1 to 5:1 for gradient elution) was carried out to obtain 0.90 g of a yellow solid with the yield of 71.4%.

4-(3-(2-chloro-44(2S,6R)-2,6-dimethylmorpholinyl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (IV-3)

(22) An intermediate 4 (0.95 g, 3.19 mmol), the compound III-3 (0.90 g, 3.18 mmol) and PyBOP (1.99 g, 3.82 mmol) were added into a 50 mL three-necked flask, and 20 mL of DMF was added and stirred to be dissolved. Then DIEA (1.84 mL, 11.14 mmol) was added and stirred at 25° C. for reaction for 6 to 8 hours. TLC (dichloromethane:methanol=20:1) was adopted to detect that the raw material 111-3 was completely reacted, and a reaction solution was poured into 60 mL of water to precipitate a yellow solid. Suction filtration was carried out, and a filter cake was washed with 20 mL of water and dried to obtain a crude product. Column chromatography purification (dichloromethane:methanol=100:1 to 40:1 for gradient elution) was carried out to obtain 1.02 g of a yellow solid with the yield of 57.0%. m.p. 149-152° C. .sup.1HNMR (300 MHz, CDCl.sub.3) δ (ppm): 11.03 (1H, s, CONH), 8.51-8.46 (1H, m, ArH), 7.83-7.70 (3H, m, ArH), 7.46-7.35 (2H, m, ArH), 7.12-7.04 (1H, m, ArH), 4.45 (1H, s, 0.5×CH.sub.2), 4.33 (1H, s, 0.5×CH.sub.2), 4.30 (2H, s, CH.sub.2), 3.92-3.67 (6H, m, 2×CH.sub.2, 2×CH), 2.78-2.74 (4H, m, 2×CH.sub.2), 1.27-1.23 (6H, m, 2CH.sub.3).

4-(3-(2-(2-aminopyrimidin-5-yl)-4-((2S,6R)-2,6-dimethylmorpholinyl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidin-7-formyl)-4-fluorobenzyl)phthalazin-1(2H)-one (I-12)

(23) With the compound IV-3 (300 mg, 0.53 mmol) and the compound V-1 (177 mg, 0.80 mmol) as raw materials, operation was the same as the method of 1-2, and column chromatography purification (dichloromethane:methanol=100:1 to 20:1 for gradient elution) was carried out to obtain 160 mg of a light-yellow solid with the yield of 48.6%. m.p. 204-206° C. .sup.1HNMR (300 MHz, DMSO-d.sub.6) δ (ppm): 12.60 (1H, s, CONH), 9.03 (1H, s, ArH), 8.94 (1H, s, ArH), 8.25 (1H, t, J=6.9 Hz, ArH), 8.00-7.76 (3H, m, ArH), 7.50-7.42 (2H, m, ArH), 7.27 (1H, t, J=9.0 Hz, ArH), 7.14 (2H, s, NH.sub.2), 4.68 (1H, s, 0.5×CH.sub.2), 4.35-4.32 (3H, m, 1.5×CH.sub.2), 3.95-3.79 (3H, m, 1.5×CH.sub.2), 3.69-3.62 (2H, m, 2CH), 3.42-3.38 (1H, m, 0.5×CH.sub.2), 2.74-2.58 (4H, m, 2×CH.sub.2), 1.14 (3H, d, J=6.2 Hz, CH.sub.3), 1.09 (3H, d, J=6.1 Hz, CH.sub.3). .sup.13CNMR (75 MHz, DMSO-d.sub.6) δ (ppm): 164.42, 164.13, 163.38, 160.31, 159.88, 159.34, 157.74, 154.81, 144.81, 134.74, 133.41, 132.07, 131.50, 129.10, 127.89, 126.07, 125.43, 123.53, 119.45, 116.21, 115.82, 112.38, 70.95, 52.63, 46.49, 43.75, 36.47, 26.15, 18.59. HRMS (ESI): m/z [M+H].sup.+. Calcd for C.sub.33H.sub.32FN.sub.9O.sub.3: 622.2685; Found: 622.2691.