Synthesis and Anti-tumor Activity Evaluation of a Series of 2- or 4-Substituted Pyrido[3,2-d]pyrimidines as Nonclassical Antifolates †

doi:10.7503/cjcu20190108

Abstract

Abstract:

Using compounds 2,4-diamino-6-(4-methylphenethyl)pyrido [3,2-d] pyrimidine(wm-5b) and 2,4-diaminopyrido [3,2-d] pyrimidine as the lead compounds, a series of 2- or 4-substituted pyrido[3,2-d]pyrimidines was synthesized as potential nonclassical antifolates by means of microwave efficiently. Besides, the effects of different substituted positions which provides more theoretical basis for the design and synthesis of nonclassical antifolates can be explored. The target compounds were characterized by ¹H NMR, ¹³C NMR as well as MS. All compounds showed certain anti-tumor activity[compound 6b, the most potent one, has IC₅₀ values of (4.09±0.48) μmol/L against HL-60 cells, IC₅₀value of (17.99±7.20) μmol/L against A549 cells and IC₅₀ value of (14.52±4.74) μmol/L against HCT116 cells], while some of them exhibited certain inhibition of recombinant human DHFR(rhDHFR). Some compounds and the lead were applied to molecular docking of the crystal structure of dihydrofolate reductase, and the activity results and structure-activity relationship were explained at the molecular level.

Key words: Nonclassical antifolate, Pyrido[3,2-d]pyrimidine derivative, Antitumor activity, Inhibition of dihydrofolate reductase

CLC Number:

O626

TrendMD:

FANG Fang,XUE Liangmin,CONG Jing,TIAN Chao,WANG Xiaowei,LIU Junyi,ZHANG Zhili. Synthesis and Anti-tumor Activity Evaluation of a Series of 2- or 4-Substituted Pyrido[3,2-d]pyrimidines as Nonclassical Antifolates ^†[J]. Chem. J. Chinese Universities, 2019, 40(10): 2111.

Figures/Tables 7

References 22

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Compd.
4	(E)-2-Chloro-6-(4-methylstyryl)pyrido[3,2-d]pyrimidin-4-amine				8.54(s, 1H), 8.44(s, 1H), 8.06(d,J=16.1 Hz, 1H), 7.98(s, 2H), 7.59(d, J=8.1 Hz, 2H), 7.37(d, J=16.1 Hz, 1H), 7.26(d, J=8.0 Hz, 2H), 2.34(s, 3H)			163.60, 156.86, 154.69, 145.23, 138.97, 135.55, 135.27, 133.95, 130.36, 130.01, 128.58, 127.70, 126.08, 21.42
5a	(E)-N²-(3-Chloro-4-fluorophenyl)-6-(4-methylstyryl)pyrido[3,2-d]pyrimidine-2,4-diamine				9.39(s, 1H), 8.29(dd,J=6.9, 2.5 Hz, 1H), 7.88(m, J=10.0 Hz, 1H), 7.85(d, J=2.8 Hz, 1H), 7.83(s, 1H), 7.7(d, J=8.6 Hz, 2H),7.71(s, 1H), 7.57(d, J=7.9 Hz, 2H), 7.33(d, J=2.8 Hz, 1H), 7.31—7.27(m, 1H), 7.24(d, J=7.8 Hz,2H), 2.34(s, 3H)			162.25, 156.93, 153.20, 150.90(d, J=21.1 Hz), 150.55, 145.33, 139.04, 138.25, 134.33, 133.68, 132.96, 129.92, 129.02, 127.69, 127.36, 126.80, 119.95, 119.34(d, J=17.1 Hz), 116.79(d, J=21.1 Hz), 21.37
5b	(E)-6-(4-Methylstyryl)-N²-(3,4,5-trimethoxyphenyl)pyrido[3,2-d]pyrimidine-2,4-diamine				9.04(s, 1H), 7.85(d,J=16.3 Hz, 1H), 7.82(d, J=8.8 Hz, 1H), 7.76(s, 1H), 7.70(d, J=28.3 Hz, 2H), 7.57(d, J=8.0 Hz, 2H), 7.40(s, 2H), 7.31(d, J=16.2 Hz, 1H), 7.24(d, J=7.9 Hz, 2H), 3.79(s, 6H), 3.62(s, 3H), 2.34(s, 3H)			162.09, 157.14, 153.00, 150.22, 138.28, 137.66, 134.38, 133.61, 132.72, 132.35, 129.95, 128.93, 127.63, 127.36, 126.92, 97.19, 60.53, 56.02, 21.33
5c	(E)-4-((4-Amino-6-(4-methylstyryl)pyrido[3,2-d]pyrimidin-2-yl)amino)benzenesulfonamide				9.61(s, 1H), 8.10(d,J=8.7 Hz, 2H),7.90(d, J=16.3 Hz, 2H), 7.84(d, J=10.1 Hz, 2H), 7.79(s, 1H), 7.71(d, J=8.8 Hz, 2H), 7.58(d, J=7.9 Hz, 2H), 7.33(d, J=16.2 Hz, 1H), 7.25(d, J=7.9 Hz, 2H), 7.17(s, 2H), 2.34(s, 3H)			162.33, 156.93, 150.81, 144.82, 138.38, 135.99, 134.33, 133.84, 133.13, 129.95, 129.04, 127.76, 127.41, 126.80, 118.30, 70.34, 21.40
7a	(E)-2-Chloro-N-(3-chloro-4-fluorophenyl)-6-(4-methylstyryl)pyrido[3,2-d]pyrimidin-4-amine				10.45(s, 1H), 8.27(dd,J=6.8, 2.6 Hz, 1H), 8.17(d, J=16.2 Hz, 1H), 8.11(t, J=5.9 Hz, 2H), 8.01(m, J=9.0, 4.2, 2.7 Hz, 1H), 7.64(d, J=8.0 Hz, 2H), 7.55(t, J= 9.1 Hz, 1H), 7.47(d, J=16.2 Hz, 1H), 7.29(d, J=7.9 Hz, 2H), 2.36(s, 3H)			158.66, 155.79, 155.61(s, J=227 Hz), 145.33, 139.22, 136.50, 135.75, 135.57, 133.84, 130.49, 130.07, 128.93, 127.84, 126.08, 124.70, 123.57, 123.50, 117.39(d, J=25 Hz), 21.45
Compd.		Name		¹H NMR(400 MHz, DMSO-d₆), δ		¹³C NMR(101 MHz, DMSO-d₆), δ
7b		(E)-2-Chloro-6-(4-methylstyryl)-N-(3,4,5-trimethoxyphenyl)pyrido[3,2-d]pyrimidin-4-amine		10.18(s, 1H), 8.11(d, J=16.5 Hz, 2H), 8.07(s, 1H), 7.63(d, J=8.0 Hz, 2H), 7.49(s, 2H), 7.45(d, J=16.3 Hz, 1H), 7.28(d, J=8.0 Hz, 2H), 3.85(s, 6H), 3.71(s, 3H), 2.35(s, 3H)		158.37, 155.99, 155.43, 153.08, 145.19, 139.20, 136.26, 135.66, 134.97, 134.22, 133.83, 130.55, 130.03, 128.45, 127.80, 126.22, 100.67, 60.64, 56.36, 21.45
7c		(E)-4-((2-Chloro-6-(4-methylstyryl)pyrido[3,2-d]pyrimidin-4-yl)amino)benzenesulfonamide		10.55(s, 1H), 8.22—8.13(m, 5H), 7.91(d,J=8.8 Hz, 2H), 7.65(d, J=8.0 Hz, 2H), 7.50(d, J=16.2 Hz, 1H), 7.37(s, 2H), 7.30(d, J=8.0 Hz, 2H), 2.36(s, 3H)		158.73, 155.75, 145.50, 141.26, 140.07, 139.32, 136.61, 135.83, 133.83, 130.55, 130.08, 128.95, 127.86, 126.86, 126.12, 122.55, 21.45
8a		(E)-N⁴-(3-Chloro-4-fluorophenyl)-6-(4-methylstyryl)pyrido[3,2-d]pyrimidine-2,4-diamine		9.79(s, 1H), 8.46(dd,J=6.8, 2.7 Hz, 1H), 8.21(m, J=9.1, 4.3, 2.7 Hz, 1H), 7.91(d, J=16.3 Hz, 1H), 7.88(d, J=8.8 Hz, 1H), 7.68(d, J=8.7 Hz, 1H), 7.62(d, J=8.1 Hz, 2H), 7.45(t, J=9.1 Hz, 1H), 7.38(d, J=16.3 Hz, 1H), 7.28(d, J=8.0 Hz, 2H), 6.73(s, 2H), 2.37(s, 3H)		160.82, 157.73, 154.72, 152.31(d, J=241 Hz), 149.47, 146.57, 138.53, 136.92, 134.35, 132.94, 132.82, 129.93, 127.70, 127.35, 126.90, 122.85, 121.80, 119.47(d, J=18.2 Hz), 116.94(d, J=21.7 Hz), 21.35
8b		(E)-6-(4-Methylstyryl)-N⁴-(3,4,5-trimethoxyphenyl)pyrido[3,2-d]pyrimidine-2,4-diamine		9.49(s, 1H), 7.88(d,J=8.8 Hz, 1H), 7.83(d, J=16.4 Hz, 1H), 7.64(d, J=8.7 Hz, 1H), 7.60(d, J=8.0 Hz, 2H), 7.56(s, 2H), 7.36(d, J=16.4 Hz, 1H), 7.25(d, J=8.0 Hz, 2H), 6.63(s, 2H), 3.86(s, 6H), 3.67(s, 3H), 2.34(s, 3H)		160.23, 157.61, 153.14, 149.30, 146.62, 138.28, 135.56, 134.35, 133.76, 133.01, 132.56, 129.93, 127.86, 127.35, 127.17, 99.21, 60.59, 56.48, 21.39
8c		(E)-4-((2-Amino-6-(4-methylstyryl)pyrido[3,2-d]pyrimidin-4-yl)amino)benzenesulfonamide		9.88(s, 1H), 8.39(d,J=8.4 Hz, 2H), 7.91(d, J=11.3 Hz, 1H), 7.86(d, J=18.3 Hz, 2H), 7.81(s, 1H), 7.68(d, J=9.0 Hz, 1H), 7.61(d, J=8.0 Hz, 2H), 7.38(d, J=16.3 Hz, 1H), 7.31(s, 2H), 7.26(d, J=7.6 Hz), 6.78(s, 2H), 2.35(s, 3H)		160.14, 157.69, 149.57, 146.95, 142.66, 138.36, 138.26, 134.34, 133.21, 132.91, 129.95, 127.73, 127.68, 127.40, 126.95, 126.79, 120.69, 21.40
13a		2-Chloro-N-(3-chloro-4-fluorophenyl)-6-methylpyrido[3,2-d]pyrimidin-4-amine		10.46(s, 1H), 8.29—8.23(m, 1H), 8.08(d,J=8.6 Hz, 1H), 7.99(m, J=9.0, 4.3, 2.7 Hz, 1H), 7.85(d, J=8.6 Hz, 1H), 7.52(t, J=9.1 Hz, 1H), 2.78(s, 3H)		159.30, 158.56, 155.61(d,J=243 Hz), 153.15, 144.92, 135.63(d, J=3.0 Hz), 135.38, 130.55, 129.79, 124.39, 123.20, 119.43(d, J=18.5 Hz), 117.14(d, J=21.7 Hz), 24.93
13b		2-Chloro-6-methyl-N-(3,4,5-trimethoxyphenyl)pyrido[3,2-d]pyrimidin-4-amine		10.15(s, 1H, NH-4), 8.06(d,J=8.5 Hz, 1H), 7.83(d, J=8.6 Hz, 1H), 7.52(s, 2H), 3.84(s, 6H), 3.70(s, 3H), 2.77(s, 3H)		159.07, 158.23, 155.80, 152.98, 144.68, 135.37, 134.71, 134.33, 130.36, 129.88, 100.25, 60.55, 56.49, 24.83
13c		4-((2-Chloro-6-methylpyrido[3,2-d]pyrimidin-4-yl)amino)benzenesulfonamide		10.51(s, 1H), 8.13(d,J=8.8 Hz, 2H), 8.10—8.04(m, 1H), 7.87(d, J=8.8 Hz, 2H), 7.83(m, J=5.4, 3.1 Hz, 1H), 7.33(s, 2H), 2.77(s, 3H)		159.49, 158.69, 155.67, 145.06, 141.29, 139.92, 135.45, 130.68, 129.85, 126.80, 122.35, 24.89
6a		N²-(3-Chloro-4-fluorophenyl)-6-(4-methylphenethyl)pyrido[3,2-d]pyrimidine-2,4-diamine		9.32(s, 1H), 8.28(d,J=6.7 Hz, 1H), 7.84(d, J=9.1 Hz, 1H), 7.70(d, J=8.5 Hz, 1H), 7.58(s, 2H), 7.52(s, J=8.7 Hz, 1H), 7.29(t, J=9.1 Hz, 1H), 7.14(d, J=7.6 Hz, 2H), 7.07(d, J=7.6 Hz, 2H), 3.11(d, J=6.6 Hz, 2H), 3.07(d, J=5.7 Hz, 2H), 2.25(s, 3H)		162.14, 156.81, 156.30, 153.20(d, J=238 Hz), 150.82, 144.85, 139.23, 138.90, 135.10, 133.63, 129.27, 128.77, 128.73, 128.42, 119.78, 119.00(d, J=20.9, 12.1 Hz), 116.77(d, J=21.4 Hz), 34.57, 21.29, 14.23
6b		6-(4-Methylphenethyl)-N²-(3,4,5-trimethoxyphenyl)pyrido[3,2-d]pyrimidine-2,4-diamine		8.96(s, 1H), 7.68(d,J=8.5 Hz, 1H), 7.48(s, 1H), 7.45(d, J=11.3 Hz, 2H), 7.42(s, 2H), 7.12(d, J=7.9 Hz, 2H), 7.05(d, J=7.9 Hz, 2H), 3.79(s, 6H), 3.62(s, 3H), 3.09(t, 2H), 3.05(t, 2H), 2.23(s, 3H)		161.96, 157.13, 155.84, 152.97, 145.03, 138.85, 137.94, 135.07, 133.66, 132.14, 129.24, 128.69, 128.61, 128.35, 97.02, 60.54, 56.10, 34.65, 21.01, 18.99
Compd.		Name	¹H NMR(400 MHz, DMSO-d₆), δ				¹³C NMR(101 MHz, DMSO-d₆), δ
6c		4-((4-Amino-6-(4-methylphenethyl)pyrido[3,2-d]pyrimidin-2-yl)amino)benzenesulfonamide	9.57(s, 1H), 8.12(d,J=8.8 Hz, 2H), 7.78(d, J=8.6 Hz, 1H), 7.71(d, J=8.8 Hz, 2H), 7.64(d, J=10.2 Hz, 2H), 7.58(d, J=8.6 Hz, 1H), 7.18(d, J=9.7 Hz, 4H), 7.10(d, J=7.9 Hz, 2H), 3.15(t, 2H), 3.10(t, 2H), 2.28(s, 3H)				162.19, 156.80, 156.57, 144.96, 144.88, 138.89, 135.78, 135.12, 133.79, 129.29, 128.83, 128.75, 128.41, 126.78, 118.08, 34.52, 31.75, 21.09
9a		N⁴-(3-Chloro-4-fluorophenyl)-6-(4-methylphenethyl)pyrido[3,2-d]pyrimidine-2,4-diamine	9.62(s, 1H), 8.41(dd,J=6.8, 2.5 Hz, 1H), 8.18—8.09(m, 1H), 7.61(d, J=8.6 Hz, 1H), 7.53(d, J=8.6 Hz, 1H), 7.43(t, J=9.1 Hz, 1H), 7.18(d, J=7.8 Hz, 2H), 7.10(d, J=7.8 Hz, 2H), 6.62(s, 2H), 3.23—3.14(t, 2H), 3.13—3.05(t, 2H), 2.27(s, 3H)				159.96, 157.63, 155.15, 154.63(d, J=241 Hz), 152.22, 146.03, 138.90, 136.93, 135.11, 132.96, 129.27, 128.73, 127.11, 122.64, 121.57, 119.45(d, J=18.2 Hz), 116.91(d, J=21.6 Hz), 36.64, 30.39, 22.03
9b		6-(4-Methylphenethyl)-N4-(3,4,5-trimethoxyphenyl)pyrido[3,2-d]pyrimidine-2,4-diamine	9.34(s, 1H), 7.59(d,J=8.5 Hz, 1H), 7.52(d, 1H), 7.50(s, 2H), 7.17(d, J=6.9 Hz, 2H), 7.09(d, J=7.4 Hz, 2H), 6.53(s, 2H), 3.86(s, 6H), 3.67(s, 3H), 3.17(t, 2H), 3.09(t, 2H), 2.26(s, 3H)				160.07, 157.54, 154.94, 153.15, 145.91, 138.84, 135.56, 135.15, 133.67, 133.00, 129.28, 128.76, 127.30, 98.98, 60.59, 56.47, 34.86, 30.48, 21.10
9c		4-((2-Amino-6-(4-methylphenethyl)pyrido[3,2-d]pyrimidin-4-yl)amino)benzenesulfonamide	9.71(s, 1H), 8.31(d,J=8.5 Hz,2H), 7.80(d, J=8.6 Hz, 2H), 7.62(d, J=8.6 Hz, 1H), 7.54(d, J=8.7 Hz, 1H), 7.31(s, 2H), 7.17(d, J=7.6 Hz, 2H), 7.09(d, J=7.5 Hz, 2H), 6.69(s, 2H), 3.17(t, 2H), 3.09(t, 2H), 2.25(s, 3H)				159.79, 157.48, 155.27, 145.56, 142.28, 138.72, 137.99, 135.10, 132.98, 129.29, 128.95, 128.77, 127.08, 126.79, 120.32, 34.55, 29.48, 21.12
12a		N²-(3-Chloro-4-fluorophenyl)-6-methylpyrido[3,2-d]pyrimidine-2,4-diamine	9.34(s, 1H), 8.32(dd,J=6.9, 2.5 Hz, 1H), 7.91—7.83(m, 1H), 7.75(d, J=8.6 Hz, 1H), 7.57(d, J=8.6 Hz, 1H), 7.54(s, 2H), 7.32(t, J=9.1 Hz, 1H), 2.62(s, 3H)				162.06, 156.74, 153.43, 153.31(d, J=238 Hz), 144.66, 139.27, 133.77, 129.20, 128.40, 119.74(d, J=29 Hz), 118.99(d, J=23.6 Hz), 116.88, 116.67, 24.44
12b		6-Methyl-N²-(3,4,5-trimethoxyphenyl)pyrido[3,2-d]pyrimidine-2,4-diamine	8.96(s, 1H), 7.74(d,J=8.5 Hz, 1H), 7.56(d, J=8.6 Hz, 1H), 7.45(s, 2H), 7.44(s, 2H), 3.83(s, 6H), 3.66(s, 3H), 2.62(s, 3H)				161.87, 157.06, 152.97, 144.88, 137.95, 133.81, 132.14, 129.06, 128.31, 97.03, 60.58, 56.13, 24.43
12c		4-((4-Amino-6-methylpyrido[3,2-d]pyri-midin-2-yl)amino)benzenesulfonamide	10.95(s, 1H), 9.27(d,J=97.5 Hz, 2H), 7.95(d, J=8.6 Hz, 1H), 7.87(m, J=20.7, 8.9 Hz, 4H), 7.77(d, J=8.6 Hz, 1H), 7.38(s, 2H), 2.65(s, 3H)				162.93, 156.57, 153.04, 150.33, 140.90, 139.87, 131.09, 127.17, 126.99, 126.40, 121.56, 24.07
14a		N⁴-(3-Chloro-4-fluorophenyl)-6-methylpyrido[3,2-d]pyrimidine-2,4-diamine	9.68(s, 1H), 8.43(dd,J=6.8, 2.3 Hz, 1H), 8.18—8.04(m, 1H), 7.62(d, J=8.6 Hz, 1H), 7.53(d, J=8.6 Hz, 1H), 7.41(t, J=9.1 Hz, 1H), 6.58(s, 2H), 2.64(s, 3H)				159.95, 157.58, 154.55, 152.14(d, J=236 Hz), 145.97, 137.09, 133.11, 129.26, 127.15, 122.54, 121.47, 119.40(d, J=18.2 Hz), 116.87(d, J=21.5 Hz), 24.29
14b		6-Methyl-N⁴-(3,4,5-trimethoxyphenyl)pyrido[3,2-d]pyrimidine-2,4-diamine	9.37(s, 1H), 7.60(d,J=8.6 Hz, 1H), 7.53(s, 2H), 7.51(d, J=8.6 Hz, 1H), 6.51(s, 2H), 3.85(s, 6H), 3.66(s, 3H), 2.63(s, 3H)				159.99, 157.44, 153.12, 151.85, 145.66, 135.67, 133.53, 133.08, 129.09, 127.29, 98.78, 60.55, 56.40, 24.72
14c		4-((2-Amino-6-methylpyrido[3,2-d]pyri-midin-4-yl)amino)benzenesulfonamide	9.78(s, 1H, NH-4), 8.34(d, J=8.8 Hz, 2H), 7.81(d, J=8.8 Hz, 2H), 7.65(d, J=8.6 Hz, 1H), 7.56(d, J=8.6 Hz, 1H), 7.32(s, 2H), 6.65(s, 2H), 2.65(s, 3H)				159.90, 157.44, 152.29, 146.07, 142.63, 137.96, 133.10, 129.39, 127.08, 126.76, 120.46, 24.32

Compd.
4	(E)-2-Chloro-6-(4-methylstyryl)pyrido[3,2-d]pyrimidin-4-amine				8.54(s, 1H), 8.44(s, 1H), 8.06(d,J=16.1 Hz, 1H), 7.98(s, 2H), 7.59(d, J=8.1 Hz, 2H), 7.37(d, J=16.1 Hz, 1H), 7.26(d, J=8.0 Hz, 2H), 2.34(s, 3H)			163.60, 156.86, 154.69, 145.23, 138.97, 135.55, 135.27, 133.95, 130.36, 130.01, 128.58, 127.70, 126.08, 21.42
5a	(E)-N²-(3-Chloro-4-fluorophenyl)-6-(4-methylstyryl)pyrido[3,2-d]pyrimidine-2,4-diamine				9.39(s, 1H), 8.29(dd,J=6.9, 2.5 Hz, 1H), 7.88(m, J=10.0 Hz, 1H), 7.85(d, J=2.8 Hz, 1H), 7.83(s, 1H), 7.7(d, J=8.6 Hz, 2H),7.71(s, 1H), 7.57(d, J=7.9 Hz, 2H), 7.33(d, J=2.8 Hz, 1H), 7.31—7.27(m, 1H), 7.24(d, J=7.8 Hz,2H), 2.34(s, 3H)			162.25, 156.93, 153.20, 150.90(d, J=21.1 Hz), 150.55, 145.33, 139.04, 138.25, 134.33, 133.68, 132.96, 129.92, 129.02, 127.69, 127.36, 126.80, 119.95, 119.34(d, J=17.1 Hz), 116.79(d, J=21.1 Hz), 21.37
5b	(E)-6-(4-Methylstyryl)-N²-(3,4,5-trimethoxyphenyl)pyrido[3,2-d]pyrimidine-2,4-diamine				9.04(s, 1H), 7.85(d,J=16.3 Hz, 1H), 7.82(d, J=8.8 Hz, 1H), 7.76(s, 1H), 7.70(d, J=28.3 Hz, 2H), 7.57(d, J=8.0 Hz, 2H), 7.40(s, 2H), 7.31(d, J=16.2 Hz, 1H), 7.24(d, J=7.9 Hz, 2H), 3.79(s, 6H), 3.62(s, 3H), 2.34(s, 3H)			162.09, 157.14, 153.00, 150.22, 138.28, 137.66, 134.38, 133.61, 132.72, 132.35, 129.95, 128.93, 127.63, 127.36, 126.92, 97.19, 60.53, 56.02, 21.33
5c	(E)-4-((4-Amino-6-(4-methylstyryl)pyrido[3,2-d]pyrimidin-2-yl)amino)benzenesulfonamide				9.61(s, 1H), 8.10(d,J=8.7 Hz, 2H),7.90(d, J=16.3 Hz, 2H), 7.84(d, J=10.1 Hz, 2H), 7.79(s, 1H), 7.71(d, J=8.8 Hz, 2H), 7.58(d, J=7.9 Hz, 2H), 7.33(d, J=16.2 Hz, 1H), 7.25(d, J=7.9 Hz, 2H), 7.17(s, 2H), 2.34(s, 3H)			162.33, 156.93, 150.81, 144.82, 138.38, 135.99, 134.33, 133.84, 133.13, 129.95, 129.04, 127.76, 127.41, 126.80, 118.30, 70.34, 21.40
7a	(E)-2-Chloro-N-(3-chloro-4-fluorophenyl)-6-(4-methylstyryl)pyrido[3,2-d]pyrimidin-4-amine				10.45(s, 1H), 8.27(dd,J=6.8, 2.6 Hz, 1H), 8.17(d, J=16.2 Hz, 1H), 8.11(t, J=5.9 Hz, 2H), 8.01(m, J=9.0, 4.2, 2.7 Hz, 1H), 7.64(d, J=8.0 Hz, 2H), 7.55(t, J= 9.1 Hz, 1H), 7.47(d, J=16.2 Hz, 1H), 7.29(d, J=7.9 Hz, 2H), 2.36(s, 3H)			158.66, 155.79, 155.61(s, J=227 Hz), 145.33, 139.22, 136.50, 135.75, 135.57, 133.84, 130.49, 130.07, 128.93, 127.84, 126.08, 124.70, 123.57, 123.50, 117.39(d, J=25 Hz), 21.45
Compd.		Name		¹H NMR(400 MHz, DMSO-d₆), δ		¹³C NMR(101 MHz, DMSO-d₆), δ
7b		(E)-2-Chloro-6-(4-methylstyryl)-N-(3,4,5-trimethoxyphenyl)pyrido[3,2-d]pyrimidin-4-amine		10.18(s, 1H), 8.11(d, J=16.5 Hz, 2H), 8.07(s, 1H), 7.63(d, J=8.0 Hz, 2H), 7.49(s, 2H), 7.45(d, J=16.3 Hz, 1H), 7.28(d, J=8.0 Hz, 2H), 3.85(s, 6H), 3.71(s, 3H), 2.35(s, 3H)		158.37, 155.99, 155.43, 153.08, 145.19, 139.20, 136.26, 135.66, 134.97, 134.22, 133.83, 130.55, 130.03, 128.45, 127.80, 126.22, 100.67, 60.64, 56.36, 21.45
7c		(E)-4-((2-Chloro-6-(4-methylstyryl)pyrido[3,2-d]pyrimidin-4-yl)amino)benzenesulfonamide		10.55(s, 1H), 8.22—8.13(m, 5H), 7.91(d,J=8.8 Hz, 2H), 7.65(d, J=8.0 Hz, 2H), 7.50(d, J=16.2 Hz, 1H), 7.37(s, 2H), 7.30(d, J=8.0 Hz, 2H), 2.36(s, 3H)		158.73, 155.75, 145.50, 141.26, 140.07, 139.32, 136.61, 135.83, 133.83, 130.55, 130.08, 128.95, 127.86, 126.86, 126.12, 122.55, 21.45
8a		(E)-N⁴-(3-Chloro-4-fluorophenyl)-6-(4-methylstyryl)pyrido[3,2-d]pyrimidine-2,4-diamine		9.79(s, 1H), 8.46(dd,J=6.8, 2.7 Hz, 1H), 8.21(m, J=9.1, 4.3, 2.7 Hz, 1H), 7.91(d, J=16.3 Hz, 1H), 7.88(d, J=8.8 Hz, 1H), 7.68(d, J=8.7 Hz, 1H), 7.62(d, J=8.1 Hz, 2H), 7.45(t, J=9.1 Hz, 1H), 7.38(d, J=16.3 Hz, 1H), 7.28(d, J=8.0 Hz, 2H), 6.73(s, 2H), 2.37(s, 3H)		160.82, 157.73, 154.72, 152.31(d, J=241 Hz), 149.47, 146.57, 138.53, 136.92, 134.35, 132.94, 132.82, 129.93, 127.70, 127.35, 126.90, 122.85, 121.80, 119.47(d, J=18.2 Hz), 116.94(d, J=21.7 Hz), 21.35
8b		(E)-6-(4-Methylstyryl)-N⁴-(3,4,5-trimethoxyphenyl)pyrido[3,2-d]pyrimidine-2,4-diamine		9.49(s, 1H), 7.88(d,J=8.8 Hz, 1H), 7.83(d, J=16.4 Hz, 1H), 7.64(d, J=8.7 Hz, 1H), 7.60(d, J=8.0 Hz, 2H), 7.56(s, 2H), 7.36(d, J=16.4 Hz, 1H), 7.25(d, J=8.0 Hz, 2H), 6.63(s, 2H), 3.86(s, 6H), 3.67(s, 3H), 2.34(s, 3H)		160.23, 157.61, 153.14, 149.30, 146.62, 138.28, 135.56, 134.35, 133.76, 133.01, 132.56, 129.93, 127.86, 127.35, 127.17, 99.21, 60.59, 56.48, 21.39
8c		(E)-4-((2-Amino-6-(4-methylstyryl)pyrido[3,2-d]pyrimidin-4-yl)amino)benzenesulfonamide		9.88(s, 1H), 8.39(d,J=8.4 Hz, 2H), 7.91(d, J=11.3 Hz, 1H), 7.86(d, J=18.3 Hz, 2H), 7.81(s, 1H), 7.68(d, J=9.0 Hz, 1H), 7.61(d, J=8.0 Hz, 2H), 7.38(d, J=16.3 Hz, 1H), 7.31(s, 2H), 7.26(d, J=7.6 Hz), 6.78(s, 2H), 2.35(s, 3H)		160.14, 157.69, 149.57, 146.95, 142.66, 138.36, 138.26, 134.34, 133.21, 132.91, 129.95, 127.73, 127.68, 127.40, 126.95, 126.79, 120.69, 21.40
13a		2-Chloro-N-(3-chloro-4-fluorophenyl)-6-methylpyrido[3,2-d]pyrimidin-4-amine		10.46(s, 1H), 8.29—8.23(m, 1H), 8.08(d,J=8.6 Hz, 1H), 7.99(m, J=9.0, 4.3, 2.7 Hz, 1H), 7.85(d, J=8.6 Hz, 1H), 7.52(t, J=9.1 Hz, 1H), 2.78(s, 3H)		159.30, 158.56, 155.61(d,J=243 Hz), 153.15, 144.92, 135.63(d, J=3.0 Hz), 135.38, 130.55, 129.79, 124.39, 123.20, 119.43(d, J=18.5 Hz), 117.14(d, J=21.7 Hz), 24.93
13b		2-Chloro-6-methyl-N-(3,4,5-trimethoxyphenyl)pyrido[3,2-d]pyrimidin-4-amine		10.15(s, 1H, NH-4), 8.06(d,J=8.5 Hz, 1H), 7.83(d, J=8.6 Hz, 1H), 7.52(s, 2H), 3.84(s, 6H), 3.70(s, 3H), 2.77(s, 3H)		159.07, 158.23, 155.80, 152.98, 144.68, 135.37, 134.71, 134.33, 130.36, 129.88, 100.25, 60.55, 56.49, 24.83
13c		4-((2-Chloro-6-methylpyrido[3,2-d]pyrimidin-4-yl)amino)benzenesulfonamide		10.51(s, 1H), 8.13(d,J=8.8 Hz, 2H), 8.10—8.04(m, 1H), 7.87(d, J=8.8 Hz, 2H), 7.83(m, J=5.4, 3.1 Hz, 1H), 7.33(s, 2H), 2.77(s, 3H)		159.49, 158.69, 155.67, 145.06, 141.29, 139.92, 135.45, 130.68, 129.85, 126.80, 122.35, 24.89
6a		N²-(3-Chloro-4-fluorophenyl)-6-(4-methylphenethyl)pyrido[3,2-d]pyrimidine-2,4-diamine		9.32(s, 1H), 8.28(d,J=6.7 Hz, 1H), 7.84(d, J=9.1 Hz, 1H), 7.70(d, J=8.5 Hz, 1H), 7.58(s, 2H), 7.52(s, J=8.7 Hz, 1H), 7.29(t, J=9.1 Hz, 1H), 7.14(d, J=7.6 Hz, 2H), 7.07(d, J=7.6 Hz, 2H), 3.11(d, J=6.6 Hz, 2H), 3.07(d, J=5.7 Hz, 2H), 2.25(s, 3H)		162.14, 156.81, 156.30, 153.20(d, J=238 Hz), 150.82, 144.85, 139.23, 138.90, 135.10, 133.63, 129.27, 128.77, 128.73, 128.42, 119.78, 119.00(d, J=20.9, 12.1 Hz), 116.77(d, J=21.4 Hz), 34.57, 21.29, 14.23
6b		6-(4-Methylphenethyl)-N²-(3,4,5-trimethoxyphenyl)pyrido[3,2-d]pyrimidine-2,4-diamine		8.96(s, 1H), 7.68(d,J=8.5 Hz, 1H), 7.48(s, 1H), 7.45(d, J=11.3 Hz, 2H), 7.42(s, 2H), 7.12(d, J=7.9 Hz, 2H), 7.05(d, J=7.9 Hz, 2H), 3.79(s, 6H), 3.62(s, 3H), 3.09(t, 2H), 3.05(t, 2H), 2.23(s, 3H)		161.96, 157.13, 155.84, 152.97, 145.03, 138.85, 137.94, 135.07, 133.66, 132.14, 129.24, 128.69, 128.61, 128.35, 97.02, 60.54, 56.10, 34.65, 21.01, 18.99
Compd.		Name	¹H NMR(400 MHz, DMSO-d₆), δ				¹³C NMR(101 MHz, DMSO-d₆), δ
6c		4-((4-Amino-6-(4-methylphenethyl)pyrido[3,2-d]pyrimidin-2-yl)amino)benzenesulfonamide	9.57(s, 1H), 8.12(d,J=8.8 Hz, 2H), 7.78(d, J=8.6 Hz, 1H), 7.71(d, J=8.8 Hz, 2H), 7.64(d, J=10.2 Hz, 2H), 7.58(d, J=8.6 Hz, 1H), 7.18(d, J=9.7 Hz, 4H), 7.10(d, J=7.9 Hz, 2H), 3.15(t, 2H), 3.10(t, 2H), 2.28(s, 3H)				162.19, 156.80, 156.57, 144.96, 144.88, 138.89, 135.78, 135.12, 133.79, 129.29, 128.83, 128.75, 128.41, 126.78, 118.08, 34.52, 31.75, 21.09
9a		N⁴-(3-Chloro-4-fluorophenyl)-6-(4-methylphenethyl)pyrido[3,2-d]pyrimidine-2,4-diamine	9.62(s, 1H), 8.41(dd,J=6.8, 2.5 Hz, 1H), 8.18—8.09(m, 1H), 7.61(d, J=8.6 Hz, 1H), 7.53(d, J=8.6 Hz, 1H), 7.43(t, J=9.1 Hz, 1H), 7.18(d, J=7.8 Hz, 2H), 7.10(d, J=7.8 Hz, 2H), 6.62(s, 2H), 3.23—3.14(t, 2H), 3.13—3.05(t, 2H), 2.27(s, 3H)				159.96, 157.63, 155.15, 154.63(d, J=241 Hz), 152.22, 146.03, 138.90, 136.93, 135.11, 132.96, 129.27, 128.73, 127.11, 122.64, 121.57, 119.45(d, J=18.2 Hz), 116.91(d, J=21.6 Hz), 36.64, 30.39, 22.03
9b		6-(4-Methylphenethyl)-N4-(3,4,5-trimethoxyphenyl)pyrido[3,2-d]pyrimidine-2,4-diamine	9.34(s, 1H), 7.59(d,J=8.5 Hz, 1H), 7.52(d, 1H), 7.50(s, 2H), 7.17(d, J=6.9 Hz, 2H), 7.09(d, J=7.4 Hz, 2H), 6.53(s, 2H), 3.86(s, 6H), 3.67(s, 3H), 3.17(t, 2H), 3.09(t, 2H), 2.26(s, 3H)				160.07, 157.54, 154.94, 153.15, 145.91, 138.84, 135.56, 135.15, 133.67, 133.00, 129.28, 128.76, 127.30, 98.98, 60.59, 56.47, 34.86, 30.48, 21.10
9c		4-((2-Amino-6-(4-methylphenethyl)pyrido[3,2-d]pyrimidin-4-yl)amino)benzenesulfonamide	9.71(s, 1H), 8.31(d,J=8.5 Hz,2H), 7.80(d, J=8.6 Hz, 2H), 7.62(d, J=8.6 Hz, 1H), 7.54(d, J=8.7 Hz, 1H), 7.31(s, 2H), 7.17(d, J=7.6 Hz, 2H), 7.09(d, J=7.5 Hz, 2H), 6.69(s, 2H), 3.17(t, 2H), 3.09(t, 2H), 2.25(s, 3H)				159.79, 157.48, 155.27, 145.56, 142.28, 138.72, 137.99, 135.10, 132.98, 129.29, 128.95, 128.77, 127.08, 126.79, 120.32, 34.55, 29.48, 21.12
12a		N²-(3-Chloro-4-fluorophenyl)-6-methylpyrido[3,2-d]pyrimidine-2,4-diamine	9.34(s, 1H), 8.32(dd,J=6.9, 2.5 Hz, 1H), 7.91—7.83(m, 1H), 7.75(d, J=8.6 Hz, 1H), 7.57(d, J=8.6 Hz, 1H), 7.54(s, 2H), 7.32(t, J=9.1 Hz, 1H), 2.62(s, 3H)				162.06, 156.74, 153.43, 153.31(d, J=238 Hz), 144.66, 139.27, 133.77, 129.20, 128.40, 119.74(d, J=29 Hz), 118.99(d, J=23.6 Hz), 116.88, 116.67, 24.44
12b		6-Methyl-N²-(3,4,5-trimethoxyphenyl)pyrido[3,2-d]pyrimidine-2,4-diamine	8.96(s, 1H), 7.74(d,J=8.5 Hz, 1H), 7.56(d, J=8.6 Hz, 1H), 7.45(s, 2H), 7.44(s, 2H), 3.83(s, 6H), 3.66(s, 3H), 2.62(s, 3H)				161.87, 157.06, 152.97, 144.88, 137.95, 133.81, 132.14, 129.06, 128.31, 97.03, 60.58, 56.13, 24.43
12c		4-((4-Amino-6-methylpyrido[3,2-d]pyri-midin-2-yl)amino)benzenesulfonamide	10.95(s, 1H), 9.27(d,J=97.5 Hz, 2H), 7.95(d, J=8.6 Hz, 1H), 7.87(m, J=20.7, 8.9 Hz, 4H), 7.77(d, J=8.6 Hz, 1H), 7.38(s, 2H), 2.65(s, 3H)				162.93, 156.57, 153.04, 150.33, 140.90, 139.87, 131.09, 127.17, 126.99, 126.40, 121.56, 24.07
14a		N⁴-(3-Chloro-4-fluorophenyl)-6-methylpyrido[3,2-d]pyrimidine-2,4-diamine	9.68(s, 1H), 8.43(dd,J=6.8, 2.3 Hz, 1H), 8.18—8.04(m, 1H), 7.62(d, J=8.6 Hz, 1H), 7.53(d, J=8.6 Hz, 1H), 7.41(t, J=9.1 Hz, 1H), 6.58(s, 2H), 2.64(s, 3H)				159.95, 157.58, 154.55, 152.14(d, J=236 Hz), 145.97, 137.09, 133.11, 129.26, 127.15, 122.54, 121.47, 119.40(d, J=18.2 Hz), 116.87(d, J=21.5 Hz), 24.29
14b		6-Methyl-N⁴-(3,4,5-trimethoxyphenyl)pyrido[3,2-d]pyrimidine-2,4-diamine	9.37(s, 1H), 7.60(d,J=8.6 Hz, 1H), 7.53(s, 2H), 7.51(d, J=8.6 Hz, 1H), 6.51(s, 2H), 3.85(s, 6H), 3.66(s, 3H), 2.63(s, 3H)				159.99, 157.44, 153.12, 151.85, 145.66, 135.67, 133.53, 133.08, 129.09, 127.29, 98.78, 60.55, 56.40, 24.72
14c		4-((2-Amino-6-methylpyrido[3,2-d]pyri-midin-4-yl)amino)benzenesulfonamide	9.78(s, 1H, NH-4), 8.34(d, J=8.8 Hz, 2H), 7.81(d, J=8.8 Hz, 2H), 7.65(d, J=8.6 Hz, 1H), 7.56(d, J=8.6 Hz, 1H), 7.32(s, 2H), 6.65(s, 2H), 2.65(s, 3H)				159.90, 157.44, 152.29, 146.07, 142.63, 137.96, 133.10, 129.39, 127.08, 126.76, 120.46, 24.32

Compd.	m. p./℃	MS, m/z[M+H]⁺	Compd.	m. p./℃	MS, m/z[M+H]⁺
6a	139—140	408.13	12a	172—173	304.07
6b	129—130	446.21	12b	171—172	342.15
6c	233—234	435.15	12c	>300	331.09
9a	183—184	408.13	14a	221	323.20
9b	165—166	446.21	14b	164	361.10
9c	266—267	435.15	14c	290—291	350.02

Compd.	m. p./℃	MS, m/z[M+H]⁺	Compd.	m. p./℃	MS, m/z[M+H]⁺
6a	139—140	408.13	12a	172—173	304.07
6b	129—130	446.21	12b	171—172	342.15
6c	233—234	435.15	12c	>300	331.09
9a	183—184	408.13	14a	221	323.20
9b	165—166	446.21	14b	164	361.10
9c	266—267	435.15	14c	290—291	350.02

Compd.	IC₅₀/(μmol·L^-1)			Compd.	IC₅₀/(μmol·L^-1)
Compd.	HL-60	A549	HCT116	Compd.	HL-60	A549	HCT116
6a	12.46±1.97	6.72±3.42	23.11±2.96	12b	30.75±2.17	34.91±18.30	35.36±0.08
6b	4.09±0.48	17.99±7.20	14.52±4.74	12c	39.29±5.45	62.51±10.23	>100
6c	6.57±0.17	22.21±8.99	>100	14a	15.43±582	30.91±0.28	16.51±11.23
9a	18.39±10.54	ND	ND	14b	6.20±0.60	18.86±4.48	16.67±7.10
9b	6.74±1.44	16.57±1.64	13.70±1.02	14c	65.35±12.89	41.82±7.01	39.99±6.67
9c	5.28±1.70	20.02±2.40	22.28±0.01	wm-5b	0.07±0.05	3.25±1.20	4.68±3.93
12a	20.35±2.35	24.40±4.88	32.58±3.43	MTX	0.023±0.001	0.041±0.02	0.281±4.56