Embodiment
The preparation of embodiment one: 4-(5-(1.3-benzodioxan)-3-phenyl-4,5-pyrazoline) benzsulfamide (compound 39)
Under agitation successively by 1,3-epoxy benzodioxan cinnamophenone (1.0g, 3.87mmol), ethanol (25mL), to Hydrazinobenzenesulfonamide (0.97g, 5.03mmol), acetic acid (1.0mL) joins in the round-bottomed flask of 50mL, still has fraction solids insoluble; Transferred to by flask in oil bath pan, back flow reaction 6h, TLC follow the tracks of reaction (developping agent V
acOEt: V
normal hexane=1: 2), after reaction terminates, filter, solid distilled water wash, last vacuum-drying, the solid obtained is dissolved in the purification of dehydrated alcohol recrystallization and obtains lenticular target compound.
White crystal, productive rate 66.4%.m.p.212~214℃;
1H?NMR(DMSO-d6,400MHz)δ:7.80(d,J=7.4Hz,2H,ArH),7.62(d,J=8.6Hz,2H,ArH),7.48~7.40(m,3H,ArH),7.11(d,J=8.6Hz,2H,ArH),7.04(s,2H,NH
2),6.87(d,J=7.9Hz,1H,ArH),6.78~6.75(m,2H,ArH),5.90(s,2H,CH
2),5.56(dd,J
1=4.7,J
2=4.7Hz,1H,5-H),?3.91(dd,J
1=12.4,J
2=12.0Hz,1H,4-H
b),3.19(dd,J
1=4.7,J
2=4.6Hz,1H,4-Ha).ESI-MS:422.1[M+H]
+.Anal.Calcd?for?C
22H
19N
3O
4S:C,H,N.
Embodiment two: the 4-(preparation of 5-(1.3-benzodioxan)-3-(m-aminomethyl phenyl-4,5-pyrazoline) benzsulfamide (compound 40)
Preparation method is with embodiment one.Obtain white crystal, productive rate 75.8%.m.p.187~188℃;
1H?NMR(DMSO-d6,400MHz)δ:7.64~7.56(m,4H,ArH),7.34(t,J=7.6Hz,1H,ArH),7.23(d,J=7.4Hz,1H,ArH),6.10(d,J=8.6Hz,2H,ArH),7.04(s,2H,NH
2),6.87(d,J=7.8Hz,1H,ArH),6.75(d,J=9.9Hz,2H,ArH),5.98(s,2H,CH
2),5.55(dd,J
1=4.7,J
2=4.8Hz,1H,5-H),3.91(dd,J
1=12.1,J
2=12.1Hz,1H,4-H
b),3.17(dd,J
1=4.8,J
2=4.7Hz,1H,4-Ha),2.37(s,3H,CH
3),.ESI-MS:436.1[M+H]
+.Anal.Calcd?for?C
21H
23N
3O
4S:C,H,N.
Embodiment three: the 4-(preparation of 5-(1.3-benzodioxan)-3-(m-p-methoxy-phenyl-4,5-pyrazoline) benzsulfamide (compound 41)
Preparation method is with embodiment one.Obtain white crystal, productive rate 76.6%.m.p.219~221℃;
1H?NMR(DMSO-d6,400MHz)δ:7.61(d,J=8.7Hz,2H,ArH),7.37~6.99(m,8H,ArH),6.87(d,J=7.9Hz,1H,ArH),6.76(s,2H,NH
2),6.00(s,2H,CH
2),5.56(dd,J
1=4.9,J
2=5.0Hz,1H,5-H),3.91(dd,J
1=12.2,J
2=12.0Hz,1H,4-H
b),3.82(s,3H,CH
3),3.18(dd,J
1=5.0,J
2=4.8Hz,1H,4-Ha).ESI-MS:452.1[M+H]
+.Anal.Calcd?for?C
21H
23N
3O
5S:C,H,N.
Embodiment four: the 4-(preparation of 5-(1.3-benzodioxan)-3-(o-fluorophenyl-4,5-pyrazoline) benzsulfamide (compound 42)
Preparation method is with embodiment one.Obtain white crystal, productive rate 66.5%.m.p.180~181℃;
1H?NMR(DMSO-d6,400MHz)δ:7.95(t,J=7.6Hz,1H,ArH),7.63(d,J=8.6Hz,2H,ArH),7.46(q,J=7.2Hz,1H,ArH),7.28(q,J=8.1Hz,2H,ArH),7.11(d,J=8.5Hz,2H,ArH),7.06(s,2H,NH
2),6.87(d,J=7.9Hz,1H,ArH),6.77(t,J=7.0Hz,2H,ArH),5.99(s,2H,CH
2),5.56(dd,J
1=5.0,J
2=5.0Hz,1H,5-H),4.01(dd,J
1=12.2,J
2=12.2Hz,1H,4-H
b),3.20(dd,J
1=4.6,J
2=4.6Hz,1H,4-Ha).ESI-MS:440.1[M+H]
+.Anal.Calcd?for?C
22H
18FN
3O
4S:C,H,N.
Embodiment five: the 4-(preparation of 5-(1.3-benzodioxan)-3-(m-fluorophenyl-4,5-pyrazoline) benzsulfamide (compound 43)
Preparation method is with embodiment one.Obtain white crystal, productive rate 69.8%.m.p.192~193℃;
1H?NMR(DMSO-d6,400MHz)δ:7.95(t,J=7.6Hz,1H,ArH),7.63(d,J=8.6Hz,2H,ArH),7.46(q,J=7.2Hz,1H,ArH),7.28(q,J=8.1Hz,2H,ArH),7.11(d,J=8.5Hz,2H,ArH),7.06(s,2H,NH
2),6.87(d,J=7.9Hz,1H,ArH),6.77(t,J=7.0Hz,2H,ArH),5.99(s,2H,CH
2),5.56(dd,J
1=5.0,J
2=5.0Hz,1H,5-H),4.01(dd,J
1=12.2,J
2=12.2Hz,1H,4-H
b),3.20(dd,J
1=4.6,J
2=4.6Hz,1H,4-Ha).ESI-MS:440.1[M+H]
+.Anal.Calcd?for?C
22H
18FN
3O
4S:C,H,N.
Embodiment six: the 4-(preparation of 5-(1.3-benzodioxan)-3-(p-ethoxyl phenenyl-4,5-pyrazoline) benzsulfamide (compound 44)
Preparation method is with embodiment one.Obtain white crystal, productive rate 77.6%.m.p.182~184℃;
1H?NMR(DMSO-d6,400MHz)δ:7.71(d,J=8.6Hz,2H,ArH),7.60(d,J=8.7Hz,2H,ArH),7.08~6.98(m,6H,ArH),6.86(d,J=7.8Hz,1H,ArH),6.74(s,2H,NH
2),5.98(s,2H,CH
2),5.50(dd,J
1=4.9,J
2=5.0Hz,1H,5-H),4.05(q,J=6.9Hz,2H,CH
2),3.87(dd,J
1=12.1,J
2=11.8Hz,1H,4-H
b),3.13(dd,J
1=5.0,J
2=4.8Hz,1H,4-Ha),1.34(t,J=6.9Hz,3H,CH
3).ESI-MS:466.1[M+H]
+.Anal.Calcd?for?C
24H
23N
3O
5S:C,H,N.
Embodiment seven: the 4-(preparation of 5-(1.3-benzodioxan)-3-(p-p-methoxy-phenyl-4,5-pyrazoline) benzsulfamide (compound 45)
Preparation method is with embodiment one.Obtain white crystal, productive rate 76.3%.m.p.200~201℃;
1H?NMR(DMSO-d6,400MHz)δ:7.73(d,J=8.8Hz,2H,ArH),7.59(d,J=8.7Hz,2H,ArH),7.08~7.01(m,6H,ArH?and?NH
2),6.87(d,J=7.9Hz,1H,ArH),6.77~6.73(m,2H,ArH),5.98(s,2H,CH
2),5.51(dd,J
1=5.0,J
2=5.1Hz,1H,5-H),3.88(dd,J
1=12.0,J
2=11.9Hz,1H,4-H
b),3.80(s,3H,OCH
3),3.15(dd,J
1=5.1,J
2=5.0Hz,1H,4-Ha).ESI-MS:452.1[M+H]
+.Anal.Calcd?for?C
23H
21N
3O
5S:C,H,N.
Embodiment eight: the 4-(preparation of 5-(1.3-benzodioxan)-3-(2.4-Dimethoxyphenyl-4,5-pyrazoline) benzsulfamide (compound 46)
Preparation method is with embodiment one.Obtain white crystal, productive rate 72.9%.m.p.180~182℃;
1H?NMR(DMSO-d6,400MHz)δ:7.60(d,J=8.6Hz,2H,ArH),7.35(d,J=7.9Hz,1H,ArH),7.17(s,1H,ArH),7.10~7.03(m,5H,ArH?and?NH
2),6.87(d,J=7.7Hz,1H,ArH),6.77~6.74(m,2H,ArH),6.98(s,2H,CH
2),5.48(dd,J
1=4.8,J
2=4.8Hz,1H,5-H),3.98(dd,J
1=12.0,J
2=11.8Hz,1H,4-H
b),3.20(dd,J
1=4.9,J
2=4.8Hz,1H,4-Ha),2.67(s,3H,CH
3),2.31(s,3H,CH
3).ESI-MS:450.1[M+H]
+.Anal.Calcd?for?C
24H
23N
3O
4S:C,H,N.
Embodiment nine: the 4-(preparation of 5-(1.3-benzodioxan)-3-(3.4-Dimethoxyphenyl-4,5-pyrazoline) benzsulfamide (compound 47)
Preparation method is with embodiment one.Obtain white crystal, productive rate 73.8%.m.p.179~180℃;
1H?NMR(DMSO-d6,400MHz)δ:7.60(d,J=8.0Hz,3H,ArH),7.49(d,J=7.8Hz,1H,ArH),7.21(d,J=7.8Hz,1H,ArH),7.08(d,J=8.3Hz,2H,ArH),7.02(s,2H,NH
2),6.86(d,J=7.8Hz,1H,ArH),6.75(s,2H,ArH),5.98(s,2H,CH
2),5.53(dd,J
1=4.4,J
2=4.6Hz,1H,5-H),3.88(dd,J
1=12.1,J
2=24.0Hz,1H,4-H
b),3.14(dd,J
1=4.5,J
2=4.4Hz,1H,4-Ha).ESI-MS:450.1[M+H]
+.Anal.Calcd?for?C
24H
23N
3O
4S:C,H,N.
Embodiment ten: the 4-(preparation of 5-(1.3-benzodioxan)-3-(p-iodophenyl-4,5-pyrazoline) benzsulfamide (compound 48)
Preparation method is with embodiment one.Obtain white crystal, productive rate 58.9%.m.p.212~214℃;
1H?NMR(DMSO-d6,400MHz)δ:7.82(d,J=7.9Hz,2H,ArH),7.62~7.54(m,4H,ArH),7.16(d,J=8.5Hz,2H,ArH),7.04(s,2H,NH
2),6.86(d,J=7.9Hz,1H,ArH),6.54(d,J=4.2Hz,2H,ArH),5.98(s,2H,CH
2),5.57(dd,J
1=4.7,J
2=5.1Hz,1H,5-H),3.91(dd,J
1=12.2,J
2=12.0Hz,1H,4-H
b),3.16(dd,J
1=4.9,J
2=5.0Hz,1H,4-Ha).ESI-MS:548.0[M+H]
+.Anal.Calcd?for?C
22H
18IN
3O
4S:C,H,N.
Embodiment 11: the 4-(preparation of 5-(1.3-benzodioxan)-3-(p-chloro-phenyl--4,5-pyrazoline) benzsulfamide (compound 49)
Preparation method is with embodiment one.Obtain white crystal, productive rate 53.8%.m.p.168~170℃;
1H?NMR(DMSO-d6,400MHz)δ:7.78(d,J=8.5Hz,2H,ArH),7.63(d,J=8.8Hz,2H,ArH),?7.49(d,J=8.6Hz,2H,ArH),7.07~7.13(m,4H,ArH),6.86(d,J=7.8Hz,1H,ArH),6.75(s,2H,NH
2),5.98(s,2H,CH
2),5.56(dd,J
1=5.1,J
2=5.3Hz,1H,5-H),3.90(dd,J
1=12.2,J
2=12.1Hz,1H,4-H
b),3.17(dd,J
1=5.3,J
2=5-2Hz,1H,4-Ha).ESI-MS:456.9[M+H]
+.Anal.Calcd?for?C
22H
18ClN
3O
4S:C,H,N.
The system back of the body of embodiment 12: 4-(5-(1.4-benzodioxan)-3-phenyl-4,5-pyrazoline) benzsulfamide (compound 50)
Preparation method is with embodiment one.To replace the cinnamophenone of 1.3-benzodioxan skeleton to obtain white crystal, productive rate 73.5% containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.156~157℃;
1H?NMR(DMSO-d6,400MHz)δ:7.79(d,J=6.8Hz,2H,ArH),7.61(d,J=8.9Hz,2H,ArH),7.47~7.40(m,3H,ArH),7.09(t,J=8.9Hz,2H,ArH),6.99(s,2H,NH
2),6.82(d,J=8.4Hz,1H,ArH),6.72~6.99(m,2H,ArH),5.54(dd,J
1=4.8,J
2=4.8Hz,1H,5-H),4.18(s,4H,CH
2),3.91(dd,J
1=12.1,J
2=12.0Hz,1H,4-H
b),3.17(dd,J
1=4.9,J
2=3.6Hz,1H,4-Ha).ESI-MS:435.1[M+H]
+.Anal.Calcd?for?C
23H
21N
3O
4S:C,H,N.
The preparation of embodiment 13: 4-(5-(1.4-benzodioxan)-3-(m-bromophenyl)-4,5-pyrazolines) benzsulfamide (compound 51)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 86.8% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.196~197℃;
1H?NMR(DMSO-d6,400MHz)δ:7.96(s,1H,ArH),7.74(d,J=7.9Hz,1H,ArH),7.76~7.57(m,3H,ArH),7.39(t,J=7.9Hz,1H,ArH),7.1(d,J=8.9Hz,2H,ArH),7.06(s,2H,NH
2),5.56(dd,J
1=4.88,J
2=5.0Hz,1H,5-H),4.19(s,4H,CH
2),3.88(dd,J
1=12.2,J
2=12.0Hz,1H,4-H
b),,3.18(dd,J
1=5.1,J
2=4.9Hz,1H,4-Ha).ESI-MS:515.4[M+H]
+.Anal.Calcd?for?C
23H
20BrN
3O
4S:C,H,N.
The preparation of embodiment 14: 4-(5-(1.4-benzodioxan)-3-(m-p-methoxy-phenyl)-4,5-pyrazolines) benzsulfamide (compound 52)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 70.1% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.146~147℃;
1H?NMR(DMSO-d6,400MHz)δ:7.60(d,J=8.9Hz,2H,ArH),7.34(d,J=4.9Hz,2H,ArH),7.11~7.00(m,3H,ArH),6.99(s,2H,NH
2),6.82(2,J=8.4Hz,2H,ArH),6.71~6.69(m,2H,ArH),5.54(dd,J
1=4.7,J
2=4.9Hz,1H,5-H),4.19(s,4H,CH
2),3.89(dd,J
1=12.1,J
2=12.0Hz,1H,4-H
b),3.80(s,3H,OCH
3),3.17(dd,J
1=4.9,J
2=4.8Hz,1H,4-Ha).ESI-MS:466.1[M+H]
+.Anal.Calcd?for?C
24H
23N
3O
5S:C,H,N.
The preparation of embodiment 15: 4-(5-(1.4-benzodioxan)-3-(o-fluorophenyl)-4,5-pyrazolines) benzsulfamide (compound 53)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 69.4% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.191~192℃;
1H?NMR(DMSO-d6,300MHz)δ:7.94(t,J=7.6Hz,1H,ArH),7.63(d,J=8.6Hz,2H,ArH),7.45(q,J=7.1Hz,1H,ArH),7.27(q,J=8.7Hz,2H,ArH),7.11(2,J=9.0Hz,4H,ArH),7.82(d,J=8.1Hz,4H,ArH),6.72(s,2H,NH
2),5.53(dd,J
1=4.7,J
2=4.8Hz,1H,5-H),4.19(s,4H,CH
2),3.99(dd,J
1=12.1,J
2=12.1Hz,1H,4-H
b),3.17(dd,J
1=2.5,J
2=2.2Hz,1H,4-Ha).ESI-MS:454.1[M+H]
+.Anal.Calcd?for?C
23H
20FN
3O
4S:C,H,N.
The preparation of embodiment 16: 4-(5-(1.4-benzodioxan)-3-(m-fluorophenyl)-4,5-pyrazolines) benzsulfamide (compound 54)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 68.7% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.200~201℃;
1H?NMR(DMSO-d6,300MHz)δ:7.84(q,J=5.6Hz,2H,ArH),7.60(d,J=8.9Hz,2H,ArH),7.29(t,J=8.8Hz,2H,ArH),7.08(t,J=8.9Hz,2H,ArH),6.99(s,2H,NH
2),6.82(q,2,J=8.1Hz,2H,ArH),6.70(t,J=8.0Hz,2H,ArH),5.54(dd,J
1=4.8,J
2=4.8Hz,1H,5-H),4.19(s,4H,CH
2),3.91(dd,J
1=12.1,J
2=11.9Hz,1H,4-H
b),3.16(dd,J
1=5.0,J
2=4.8Hz,1H,4-Ha).ESI-MS:454.1[M+H]
+.Anal.Calcd?for?C
23H
20FN
3O
4S:C,H,N.
The preparation of embodiment 17: 4-(5-(1.4-benzodioxan)-3-(3.5-difluorophenyl)-4,5-pyrazolines) benzsulfamide (compound 55)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 63.4% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.182~183℃;
1H?NMR(DMSO-d6,300MHz)δ:7.62(q,J=8.9Hz,2H,ArH),7.50~7.47(m,2H,ArH),7.31~7.26(m,1H,ArH),7.15(t,J=8.9Hz,2H,ArH),7.10(s,2H,NH
2),6.82(t,J=8.2Hz,2H,ArH),6.72~6.69(m,2H,ArH),5.60(dd,J
1=4.9,J
2=5.4Hz,1H,5-H),4.19(s,4H,CH
2),3.88(dd,J
1=12.1,J
2=11.9Hz,1H,4-H
b),3.19(dd,J
1=5.0,J
2=4.8Hz,1H,4-Ha).ESI-MS:472.1[M+H]
+.Anal.Calcd?for?C
23H
19F
2N
3O
4S:C,H,N.
The preparation of embodiment 18: 4-(5-(1.4-benzodioxan)-3-(p-ethoxyl phenenyl)-4,5-pyrazolines) benzsulfamide (compound 56)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 59.8% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.193~194℃;
1H?NMR(DMSO-d6,400MHz)δ:7.71(d,J=8.6Hz,2H,ArH),7.60(d,J=8.7Hz,2H,ArH),7.07~6.98(m,6H,ArH),6.81(d,J=8.0Hz,1H,ArH),6.70(s,2H,NH
2),5.48(dd,J
1=5.0,J
2=4.7Hz,1H,5-H),4.18(s,4H,CH
2),4.05(q,J=6.8Hz,2H,CH
2),3.86(dd,J
1=12.0,J
2=11.7Hz,1H,4-H
b),3.12(dd,J
1=4.7,J
2=4.6Hz,1H,4-Ha),1.34(t,J=6.9Hz,3H,CH
3).ESI-MS:480.1[M+H]
+.Anal.Calcd?for?C
25H
25N
3O
5S:C,H,N.
The preparation of embodiment 19: 4-(5-(1.4-benzodioxan)-3-(p-p-methoxy-phenyl)-4,5-pyrazolines) benzsulfamide (compound 57)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 75.6% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.207~208℃;
1H?NMR(DMSO-d6,400MHz)δ:7.73(d,J=8.8Hz,2H,ArH),7.59(d,J=8.7Hz,2H,ArH),7.08~7.01(m,6H,ArH?and?NH
2),6.87(d,J=7.9Hz,1H,ArH),6.77~6.73(m,2H,ArH),5.49(dd,J
1=5.0,J
2=5.1Hz,1H,5-H),4.19(s,4H,CH
2),3.87(dd,J
1=12.0,J
2=11.9Hz,1H,4-H
b),3.81(s,3H,OCH
3),3.13(dd,J
1=5.1,J
2=5.0Hz,1H,4-Ha).ESI-MS:466.1[M+H]
+.Anal.Calcd?for?C
24H
23N
3O
5S:C,H,N.
The preparation of embodiment 20: 4-(5-(1.4-benzodioxan)-3-(p-aminomethyl phenyl)-4,5-pyrazolines) benzsulfamide (compound 58)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 75.6% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.153~154℃;
1H?NMR(DMSO-d6,400MHz)δ:7.68(d,J=8.1Hz,2H,ArH),7.59(d,J=8.9Hz,2H,ArH),7.26(d,J=8.0Hz,2H,ArH),7.06(t,J=8.9Hz,1H,ArH),6.99(s,2H,NH
2),6.82(d,J=8.6Hz,1H,ArH),6.70~6.68(m,2H,ArH),5.51(dd,J
1=4.7,J
2=4.7Hz,1H,5-H),4.18(s,4H,CH
2),3.88(dd,J
1=12.0,J
2=11.8Hz,1H,4-H
b),3.13(dd,J
1=4.9,J
2=4.8Hz,1H,4-Ha),2.34(s,3H,CH
3).ESI-MS:450.1[M+H]
+.Anal.Calcd?for?C
24H
23N
3O
4S:C,H,N.
The preparation of embodiment 21: 4-(5-(1.4-benzodioxan)-3-(3.4-3,5-dimethylphenyl)-4,5-pyrazolines) benzsulfamide (compound 59)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 72.3% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.166~168℃;
1H?NMR(DMSO-d6,400MHz)δ:7.60(d,J=8.7Hz,3H,ArH),7.48(d,J=7.8Hz,2H,ArH),7.20(d,J=7.9Hz,1H,ArH),7.07(t,J=8.7Hz,3H,ArH),6.81(d,J=8.4Hz,1H,ArH),6.90(s,2H,NH
2),5.50(dd,J
1=4.5,J
2=4.6Hz,1H,5-H),4.19(s,4H,CH
2),3.86(dd,J
1=12.0,J
2=11.8Hz,1H,4-H
b),3.12(dd,J
1=4.6,J
2=4.5Hz,1H,4-Ha),2.27(s,3H,CH
3),2.26(s,3H,CH
3).ESI-MS:464.1[M+H]
+.Anal.Calcd?for?C
25H
25N
3O
4S:C,H,N.
The preparation of embodiment 22: 4-(5-(1.4-benzodioxan)-3-(3-methyl-4-chloro-phenyl-)-4,5-pyrazolines) benzsulfamide (compound 60)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 71.4% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.159~161℃;
1H?NMR(DMSO-d6,300MHz)δ:7.78(q,J=8.9Hz,1H,ArH),7.64~7.60(m,3H,ArH),7.48~7.40(m,1H,ArH),7.11(t,J=8.9Hz,2H,ArH),7.09(s,2H,NH
2),6.82(t,J=8.2Hz,2H,ArH),6.72~6.69(m,2H,ArH),5.54(dd,J
1=4.9,J
2=5.4Hz,1H,5-H),4.19(s,4H,CH
2),3.86(dd,J
1=12.1,J
2=11.9Hz,1H,4-H
b),3.15(dd,J
1=5.0,J
2=4.8Hz,1H,4-Ha),2.35(s,3H,CH
3),.ESI-MS:468.1[M+H]
+.Anal.Calcd?for?C
24H
22ClN
3O
4S:C,H.N.
The preparation of embodiment 23: 4-(5-(1.4-benzodioxan)-3-(p-chloro-phenyl-)-4,5-pyrazolines) benzsulfamide (compound 61)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 73.9% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.146~148℃;
1H?NMR(DMSO-d6,400MHz)δ:7.79(d,J=8.6Hz,2H,ArH),7.61(d,J=8.8Hz,2H,ArH),7.50(d,J=8.6Hz,2H,ArH),7.11~7.08(m,3H,ArH),6.82(2,J=8.0Hz,2H,ArH),6.99(s,2H,NH
2),5.55(dd,J
1=4.8,J
2=4.8Hz,1H,5-H),4.19(s,4H,CH
2),3.89(dd,J
1=7.0,J
2=12.0Hz,1H,4-H
b),3.16(dd,J
1=5.0,J
2=4.9Hz,1H,4-Ha).ESI-MS:470.1[M+H]
+.Anal.Calcd?for?C
23H
20ClN
3O
4S:C,H,N.
The preparation of embodiment 24: 4-(5-(1.4-benzodioxan)-3-(3.4-dichlorophenyl)-4,5-pyrazolines) benzsulfamide (compound 62)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 73.5% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.171~173℃;
1H?NMR(DMSO-d6,400MHz)δ:7.7.97(d,J=1.9Hz,2H,ArH),7.76~7.61(m,5H,ArH),7.24~7.06(m,4H,hrH),6.82~6.68(m,3H,ArH?and?NH
2),5.59(dd,J
1=4.8,J
2=5.0Hz,1H,5-H),4.19(s,4H,CH
2),3.89(dd,J
1=12.0,J
2=24.0Hz,1H,4-H
b),3.19(dd,J
1=5.0,J
2=4.9Hz,1H,4-Ha).ESI-MS:504.1[M+H]
+.Anal.Calcd?for?C
23H
19C1
2N
3O
4S:C,H,N.
The preparation of embodiment 25: 4-(5-(1.4-benzodioxan)-3-(p-bromophenyl)-4,5-pyrazolines) benzsulfamide (compound 63)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 63.5% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.153~154℃;
1H?NMR(DMSO-d6,300MHz)δ:7.72(d,J=8.6Hz,2H,ArH),7.61(q,J=8.8Hz,2H,ArH),7.09(t,J=8.8Hz,2H,ArH),6.99(s,2H,NH
2),6.81(2,J=8.0Hz,2H,ArH),5.55(dd,J
1=4.9,J
2=4.9Hz,1H,5-H),4.19(s,4H,CH
2),3.91(dd,J
1=12.2,J
2=12.0Hz,1H,4-H
b),3.16(dd,J
1=5.0,J
2=4.9Hz,1H,4-Ha).ESI-MS:514.0[M+H]
+.Anal.Calcd?for?C
23H
20BrN
3O
4S:C,H,N.
The preparation of embodiment 26: 4-(5-(1.4-benzodioxan)-3-(p-fluorophenyl)-4,5-pyrazolines) benzsulfamide (compound 64)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 67.8% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.195~196℃;
1H?NMR(DMSO-d6,300MHz)δ:7.84(q,J=5.6Hz,2H,ArH),7.60(d,J=8.9Hz,2H,ArH),7.29(t,J=8.8Hz,2H,ArH),7.08(t,J=8.9Hz,2H,ArH),6.99(s,2H,NH
2),6.82(q,2,J=8.1Hz,2H,ArH),6.70(t,J=8.0Hz,2H,ArH),5.54(dd,J
1=4.8,J
2=4.8Hz,1H,5-H),4.19(s,4H,CH
2),3.91(dd,J
1=12.1,J
2=11.9Hz,1H,4-H
b),3.16(dd,J
1=5.0,J
2=4.8Hz,1H,4-Ha).ESI-MS:454.1[M+H]
+.Anal.Calcd?for?C
23H
20FN
3O
4S:C,H,N.
The preparation of embodiment 27: 4-(5-(1.4-benzodioxan)-3-(2.4-difluorophenyl)-4,5-pyrazolines) benzsulfamide (compound 65)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 65.3% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.168~169℃;
1H?NMR(DMSO-d6,300MHz)δ:7.97(q,J=7.6Hz,1H,ArH),7.61(d,J=7.9Hz,2H,ArH),7.36(q,J=9.7Hz,1H,ArH),7.20(t,J=8.4Hz,2H,ArH),7.10(d,J=9.0Hz,2H,ArH),7.08(s,2H,NH
2),6.81(d,J=8.4Hz,1H,ArH),6.70(d,J=10.0Hz,2H,ArH),5.53(dd,J
1=4.5,J
2=4.7Hz,1H,5-H),4.19(s,4H,CH
2),3.98(dd,J
1=12.3,J
2=12.2Hz,1H,5-H),3.44(dd,J
1=6.8,J
2=7.4Hz,1H,4-H
b),3.14(dd,J
1=2.5,J
2=4.3Hz,1H,4-Ha).ESI-MS:472.1[M+H]
+.Anal.Calcd?for?C
23H
19F
2N
3O
4S:C,H,N.
The preparation of embodiment 28: 4-(5-(1.4-benzodioxan)-3-(p-iodophenyl)-4,5-pyrazolines) benzsulfamide (compound 66)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 72.3% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.187~189℃;
1H?NMR(DMSO-d6,300MHz)δ:7.79(d,J=8.1Hz,2H,ArH),7.63(d,J=7.6Hz,2H,ArH),7.55(d,J=8.0Hz,2H,ArH),7.11(d,J=8.6Hz,2H,ArH),6.82(2,J=8.1Hz,2H,ArH),6.70(s,2H,NH
2),5.53(dd,J
1=4.6,J
2=4.7Hz,1H,5-H),4.18(s,4H,CH
2),3.87(dd,J
1=12.4,J
2=12.0Hz,1H,4-H
b),3.13(dd,J
1=4.7,J
2=4.5Hz,1H,4-Ha).ESI-MS:562.0[M+H]
+.Anal.Calcd?for?C
23H
20IN
3O
4S:C,H,N.
The preparation of embodiment 29: 4-(5-(1.4-benzodioxan)-3-(the fluoro-4-p-methoxy-phenyl of 2-)-4,5-pyrazolines) benzsulfamide (compound 67)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 89.5% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.211~212℃;
1H?NMR(DMSO-d6,400MHz)δ:7.85(t,J=8.8Hz,1H,ArH),7.60(d,J=8.9Hz,2H,ArH),7.06(d,J=8.9Hz,3H,ArH),6.69~6.93(m,5H,ArH?and?NH
2),5.47(dd,J
1=4.9,J
2=5.0Hz,1H,5-H),4.09(s,4H,CH
2),3.94(dd,J
1=12.0,J
2=11.5Hz,1H,4-H
b),3.82(s,3H,CH
3),3.12(dd,J
1=3.6,J
2=2.64Hz,1H,4-Ha).ESI-MS:484.5[M+H]
+.Anal.Calcd?for?C
24H
22FN
3O
5S:C,H,N.
The preparation of embodiment 30: 4-(5-(1.4-benzodioxan)-3-(p-methylthio group phenyl)-4,5-pyrazolines) benzsulfamide (compound 68)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 87.3% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.202~203℃;
1H?NMR(DMSO-d6,400MHz)δ:7.71(d,J=8.4Hz,2H,ArH),7.60(d,J=9.0Hz,2H,ArH),7.32(d,J=8.5Hz,2H,ArH),6.69~7.09(m,7H,ArH?and?NH
2),5.52(dd,J
1=4.8,J
2=5.0Hz,1H,5-H),4.19(s,4H,CH
2),3.88(dd,J
1=12.0,J
2=11.9Hz,1H,4-H
b),3.15(dd,J
1=5.0,J
2=4.8Hz,1H,4-Ha),2.52(s,3H,CH
3).ESI-MS:482.6[M+H]
+.Anal.Calcd?for?C
24H
23N
3O
4S
2:C,H,N.
The preparation of embodiment 31: 4-(5-(1.4-benzodioxan)-3-(m-iodophenyl)-4,5-pyrazolines) benzsulfamide (compound 69)
Preparation method is with embodiment one.To replace obtaining tawny crystal, productive rate 87.3% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.209~210℃;
1H?NMR(DMSO-d6,400MHz)δ:8.13(s,H,ArH),7.75(d,J=8.9Hz,2H,ArH),7.61(d,J=8.8Hz,2H,ArH),7.23(t,J=7.8Hz,H,ArH),7.11(d,J=8.9Hz,2H,ArH),7.06(s,2H,NH
2),6.81(d,J=8.1Hz,H,ArH),6.68~6.70(m,2H,ArH),5.55(dd,J
1=4.8,J
2=4.9Hz,1H,5-H),4.18(s,4H,CH
2),3.87(dd,J
1=12.2,J
2=12.0Hz,1H,4-H
b),3.16(dd,J
1=5.0,J
2=4.8Hz,1H,4-Ha).ESI-MS:562.4[M+H]
+.Anal.Calcd?for?C
23H
20IN
3O
4S:C,H,N.
The preparation of embodiment 32: 4-(5-(1.4-benzodioxan)-3-(p-trifluoromethyl)-4,5-pyrazolines) benzsulfamide (compound 70)
Preparation method is with embodiment one.To replace obtaining yellow crystals, 63.3% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.216~218℃;
1H?NMR(DMSO-d6,400MHz)δ:7.97(d,J=8.1Hz,2H,ArH),7.79(d,J=8.4Hz,2H,ArH),7.63(d,J=8.9Hz,2H,ArH),7.14(d,J=8.9Hz,2H,ArH),6.70~6.83(m,3H,ArH?and?NH
2),5.61(dd,J
1=5.0,J
2=5.1Hz,1H,5-H),4.19(s,4H,CH
2),3.91~4.03(m,2H,ArH?and?4-H
b),3.21(dd,J
1=5.1,J
2=5.0Hz,1H,4-Ha).ESI-MS:504.5[M+H]
+.Anal.Calcd?for?C
24H
20F
3N
3O
4S:C,H,N.
The preparation of embodiment 33: 4-(5-(1.4-benzodioxan)-3-(p-nitrophenyl)-4,5-pyrazolines) benzsulfamide (compound 71)
Preparation method is with embodiment one.To replace obtaining red crystals, productive rate 89.6% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.219~220℃;
1H?NMR(DMSO-d6,400MHz)δ:8.27(d,J=9.0Hz,2H,ArH),7.99(d,J=8.0Hz,2H,ArH),7.65(d,J=9.9Hz,2H,ArH),7.18(d,J=8.9Hz,2H,ArH),7.09(s,2H,NH
2),6.82(d,J=8.24Hz,H,ArH),6.70~6.74(m,2H,ArH),5.65(dd,J
1=5.0,J
2=5.2Hz,1H,5-H),4.19(s,4H,CH
2),3.96(dd,J
1=12.4,J
2=12.2Hz,1H,4-H
b),3.23(dd,J
1=5.2,J
2=5.1Hz,1H,4-Ha).ESI-MS:481.5[M+H]
+.Anal.Calcd?for?C
23H
20IN
3O
4S:C,H,N.Anal.Calcd?for?C
23H
20N
4O
6S:C,H,N.
The preparation of embodiment 34: 4-(3.5-(1.3-benzodioxan)-4,5-pyrazolines) benzsulfamide (compound 72)
Preparation method is with embodiment one.Obtain white crystal, productive rate 48.6%.m.p.196~198℃;
1HNMR(DMSO-d6,400MHz)δ:7.60(d,J=8.5Hz,2H,ArH),7.43(s,1H,ArH),7.20(d,J=8.1Hz,1H,ArH),7.08(d,J=8.4Hz,2H,ArH),7.04(s,2H,NH
2),6.97(d,J=8.0Hz,1H,ArH),6.86(d,J=7.9Hz,1H,ArH),6.74(d,J=8.12Hz,2H,ArH),6.09(s,2H,CH
2),5.98(s,2H,CH
2),5.51(dd,J
1=4.6,J
2=4.8Hz,1H,5-H),3.86(dd,J
1=12.2,J
2=12.0Hz,1H,4-H
b),3.13(dd,J
1=4.6,J
2=4.5Hz,1H,4-Ha).ESI-MS:465.1[M+H]
+.Anal.Calcd?for?C
23H
19N
3O
6S:C,H,N.
The preparation of embodiment 35: 4-(3.5-(1.3-benzodioxan)-5-(1.4-benzodioxan)-4,5-pyrazolines) benzsulfamide (compound 73)
Preparation method is with embodiment one.To replace obtaining white crystal, productive rate 58.4% containing the cinnamophenone of 1.3-benzodioxan skeleton containing the cinnamophenone of 1.4-benzodioxan skeleton.m.p.186~188℃;
1H?NMR(DMSO-d6,300MHz)δ:7.58(d,J=8.9Hz,2H,ArH),7.41(s,1H,ArH),7.20(q,J=1.2Hz,1H,ArH),7.05(q,J=2.8Hz,4H,ArH),6.98(d,J=8.1Hz,1H,ArH),7.81(d,J=8.0Hz,1H,ArH),6.70(s,2H,NH
2),6.10(s,2H,CH
2),5.50(dd,J
1=4.7,J
2=4.8Hz,1H,5-H),4.19(s,4H,CH
2),3.84(dd,J
1=12.1,J
2=11.8Hz,1H,4-H
b),3.12(dd,J
1=4.9,J
2=4.7Hz,1H,4-Ha).ESI-MS:480.1[M+H]
+.Anal.Calcd?for?C
24H
21N
3O
6S:C,H,N.
Embodiment 36: the pyrazoline sulphone amide derivative anti tumor activity in vitro containing benzodioxan skeleton is studied
Adopt MTT [3-(4,5)-bis-methyl-2-thiazole-(2,5)-phenyl bromination tetrazole is blue] method measures the half-inhibition concentration (IC of pyrazoline sulphone amide derivative to human breast cancer cell (MCF-7), cervical cancer cell (HeLa), lung carcinoma cell (A549) and liver cancer cell (Hepg2)
50).
(1) preparation of nutrient solution (often liter): 1. suspension cell: DMEM cultivates one bag, powder (10.4g), new-born calf serum 100mL, penicillin solution (200,000 U/mL) 0.5mL, Streptomycin Solution (200,000 U/mL) 0.5mL, after adding tri-distilled water dissolving, with the NaHCO of 5.6%
3solution adjusts pH value to 7.2-7.4, is finally settled to 1000mL.Filtration sterilization.2. attached cell: the same, then add NaHCO
32.00g, HEPES 2.38g.
(2) preparation of D-Hanks damping fluid (often liter): NaCl 8.00g, KCl 0.40g, Na
2hPO
412H
2o0.06g, KH
2pO
40.06g, NaHCO
30.35g.Autoclaving.
(3) preparation of trypsin solution: utilizing D-Hanks damping fluid to be made into concentration is 0.5% trypsin solution.Filtration sterilization.
(4) preparation of liquid is tested: dissolved by a small amount of tri-distilled water of test sample and be made into storing solution, generally 10 times of preparation storing solutions of empirically maximum concentration.Different according to compound dissolution, available tri-distilled water directly dissolves, or with a small amount of DMSO hydrotropy, then adds tri-distilled water and dissolve.The concentration of DMSO in nutrient solution is unsuitable excessive, and in the every porocyte suspension after dosing, the final concentration of DMSO is generally no more than 0.05-0.1%.Storing solution is stored in-20 DEG C of refrigerators for subsequent use.
(5) cultivation of human breast cancer cell (MCF-7), cervical cancer cell (HeLa), lung carcinoma cell (A549) and liver cancer cell (HepG2): be adherent growth cell, cellar culture is (containing 10% calf serum, 100U/mL Streptomycin sulphate) in DMEM nutrient solution, is placed in 37 DEG C, 5%CO
2cultivate in incubator, went down to posterity once every 3-4 days.When going down to posterity, nutrient solution in former bottle is transferred in centrifuge tube, the centrifugal 5min of 1000rpm, discard original fluid, add equivalent fresh medium, piping and druming evenly, pipette appropriate in fresh culture bottle, then supplement fresh medium to original volume (nutrient solution volume is about 1/10 of culturing bottle capacity).
(6) cell incubation: the tumour cell in vegetative period of taking the logarithm, adjusts concentration of cell suspension to be (1-1.5) × 10
5individual mL
-1.In 96 well culture plates, every hole adds cell suspension 100 μ L, puts 37 DEG C, 5%CO
224h is cultivated in incubator.After cultivating 24h, add liquid by design respectively.
(7) dosing: join in each hole by test liquid respectively according to the concentration gradient of ultimate density, each concentration establishes 6 parallel holes.Experiment is divided into drug test group (adding the test medicine of different concns respectively), control group (only add nutrient solution and cell, do not add test medicine) and blank group (only add nutrient solution, do not add cell and test medicine).96 orifice plates after dosing are placed in 37 DEG C, 5%CO
248h is cultivated in incubator.The activity of positive control medicine measures according to the method for test sample.
(8) mensuration of survivaling cell: in 96 orifice plates after having cultivated 48h, every hole adds MTT 40 μ L (being made into 4mg/mL with D-Hanks damping fluid).After placing 4h at 37 DEG C, remove supernatant liquor.Every hole adds 150 μ LDMSO, and vibration 5min, makes formazan dissolving crystallized.Finally, automatic microplate reader is utilized to detect the optical density(OD) (OD value) in each hole at 570nm wavelength place.
Half-inhibition concentration (IC
50) be defined as drug level when the tumor cell survival of 50%.According to the optical density(OD) (OD value) measured, make the typical curve of inhibitory rate of cell growth, typical curve is tried to achieve the drug level of its correspondence.
The IC recorded
50be shown in Table 1.
The listed pyrazoline sulphone amide derivative containing benzodioxan skeleton of table 1 the present invention is to the suppression IC of tumour cell
50value (μm ol/mL)
a3 parallel tests, experimental result is averaged, and error is between 5%-10%
Embodiment 37: containing the pyrazoline sulphone amide derivative anti tumor activity in vitro of benzodioxan skeleton about the research suppressing MMP-2 (MMP-2) active
1. experiment material
Reagent: MMP-2 proteolytic enzyme rhMMP2,10082-HNAH, Sino Biological Inc.; Mca-PLGL-Dpa-AR-NH
2(AnaSpec, Catalog#27076), Shanghai gill biochemistry corporation,Ltd..P-aminophenylmercuric acetate (APMA) (Genmed Scientifics Inc.U.S.A, Catalog#GMS12197v.A), Sigma company produces.Other conventional chemical reagent is domestic analytical pure.
Instrument: TECAN A-5882 microplate reader (Switzerland), Other Instruments is the same.
2. experimental technique
(1) with containing the Assay Buffer of 1mM APMA by rhMMP2 protein activation, its ultimate density is made to be 100 μ g/mL.
(2) rhMMP2 albumen is hatched 1h at 37 DEG C.
(3) with Assay Buffer, the rhMMP2 albumen after activation is diluted to 0.2 μ g/mL.
(4) with Assay Buffer, substrate is diluted to 40 μMs.
(5) in 96 orifice plates, add the rhMMP2 after 50 μ L dilutions, and add the liquid 25 μ L of different concns, act on and add 25 μ L at low temperatures after 10 minutes, the substrate Mca-PLGL-Dpa-AR-NH of 40 μMs
2.
(6) fluorescence is surveyed in Ex320nm, EM450nm place, kinetic measurement 5min.
Active calculating: inhibiting rate=[(Ke-Kc)-(Ks-Kc)]/(Ke-Kc) × 100%
Ke: enzyme activity contrast Kc: blank Background control Ks: experimental value
The IC recorded
50be shown in Table 2.
The listed pyrazoline sulphone amide derivative containing benzodioxan skeleton of table 2 the present invention is to the suppression IC of matrix metalloproteinase
50value (μm ol/mL)
a3 parallel tests, experimental result is averaged, and error is between 5%-10%
Embodiment 38: containing the pyrazoline sulphone amide derivative anti tumor activity in vitro of benzodioxan skeleton about Cytotoxic research
The present invention tests new synthetic compound 39-73 to the cytotoxicity of people's renal epithelial cell (293T), cytotoxicity result as table 3, using Celecoxib as positive control.The toxicity suppressor T cell survival rate of each compound to 50% time concentration (CC
50) represent.
Experimental technique:
(1) cultivator renal epithelial cell (293T) is tending towards fusion until reach its logarithmic growth end of term cell, digests cell dispersion, be mixed with 1 × 10 with cell culture fluid with cell dissociation buffer
4the cell suspension of individual/mL.Get 96 well culture plates, in every hole, add the cell suspension of 100 μ L.Horizontally rotating culture plate gently makes cell be evenly dispersed in the surface in ware hole.
(2) be placed in containing 5%CO
2in cell culture incubator, at 37 ± 2 DEG C of temperature, cultivate 24h.Discard original fluid, every hole adds the blank liquid of 100 μ L, negative controls, positive control solution, the test sample vat liquor of 100% and 50% concentration.Often organize and at least establish 8 holes.Note: lixiviate stoste or make the serial lixiviate diluent of thinner with substratum.When adopting 0.9% sodium chloride injection lixiviate, use 2 times of concentrated substratum when diluting lixiviate.
(3) be placed in containing 5%CO
2in incubator, cultivate at 37 ± 2 DEG C of temperature.Cultivate 48h.
(4) after date between each cultivation, every hole adds MTT solution 20 μ L, is placed in containing 5%CO
2in incubator, at 37 ± 2 DEG C of temperature, cultivate 5h.
(5) discard liquid in hole, every hole adds 200 μ L DMSO respectively, and culture plate is placed 10min, and horizontal jolting makes solution colour in hole even.
(6) measure absorbancy by microplate reader, wavelength adopts 570nm.
The CC recorded
50be shown in Table 3.
The listed pyrazoline sulphone amide derivative containing benzodioxan skeleton of table 3 the present invention is to the suppression CC of 293T cell
50value (μm ol/mL)
a3 parallel tests, experimental result is averaged, and error is between 5%-10%.