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Parameter	Data
Wavelengths/Å	0.97852
Space group	P 3₂ 2 1
Cell dimensions/Å	90.870/90.870/122.820
Resolution/Å	2.90
Number of reflections	12 608
Redundancy	7.2 (7.4)
Completeness/%	94.7 (95.9)
R_merge/%^a	28.3 (53.0)
Ⅰ/σ (Ⅰ)^b	6.6 (4.0)
R/R_free/%^c	15.77 (21.96)
Bonds/Å	0.010
Angles (deg.)	1.236

Parameter

Data

Wavelengths/Å

0.97852

Space group

P 3₂ 2 1

Cell dimensions/Å

90.870/90.870/122.820

Resolution/Å

2.90

Number of reflections

12 608

Redundancy

7.2 (7.4)

Completeness/%

94.7 (95.9)

R_merge/%^a

28.3 (53.0)

Ⅰ/σ (Ⅰ)^b

6.6 (4.0)

R/R_free/%^c

15.77 (21.96)

Bonds/Å

0.010

Angles (deg.)

1.236

Parameter	Data
Wavelengths/Å	0.97852
Space group	P 3₂ 2 1
Cell dimensions/Å	90.870/90.870/122.820
Resolution/Å	2.90
Number of reflections	12 608
Redundancy	7.2 (7.4)
Completeness/%	94.7 (95.9)
R_merge/%^a	28.3 (53.0)
Ⅰ/σ (Ⅰ)^b	6.6 (4.0)
R/R_free/%^c	15.77 (21.96)
Bonds/Å	0.010
Angles (deg.)	1.236

Parameter

Data

Wavelengths/Å

0.97852

Space group

P 3₂ 2 1

Cell dimensions/Å

90.870/90.870/122.820

Resolution/Å

2.90

Number of reflections

12 608

Redundancy

7.2 (7.4)

Completeness/%

94.7 (95.9)

R_merge/%^a

28.3 (53.0)

Ⅰ/σ (Ⅰ)^b

6.6 (4.0)

R/R_free/%^c

15.77 (21.96)

Bonds/Å

0.010

Angles (deg.)

1.236

Compd.	Apearance	Yield/%	¹H NMR
2b	White solid	43	¹H NMR (400 MHz, DMSO-d₆)δ10.13(s, 1H), 9.43(s, 1H), 8.10(d, J=8.0 Hz, 2H), 7.95(s, 1H), 7.88(d, J=8.0 Hz, 2H)^[15]
3b	White solid	53	¹H NMR (400 MHz, DMSO-d₆)δ 14.09(s, 1H), 13.83(s, 1H), 8.12(d, J=8.4 Hz, 2H), 7.91(d, J=8.4 Hz, 2H)^[16]
4b	White solid	47	¹H NMR (400 MHz, DMSO-d₆)δ 8.19(d, J=8.8 Hz, 2H), 7.88(s, 2H), 3.21(d, J=1.6 Hz, 2H), 1.34(t, J=7.2 Hz, 3H)^[17]
2c	White solid	56	¹H NMR (400 MHz, DMSO-d₆)δ10.47(s, 1H), 9.37(s, 1H), 7.91(d, J=8.4 Hz, 2H), 7.70(s, 1H), 7.57(d, J=8.4 Hz, 2H)^[18]
3c	White solid	60	¹H NMR (400 MHz, DMSO-d₆)δ 13.93(s, 1H), 13.75(s, 1H), 7.92(d, J=8.4 Hz, 2H), 7.61(d, J=8.4 Hz, 2H)^[19]
4c	White solid	59	¹H NMR (400 MHz, DMSO-d₆)δ7.98(d, J=8 Hz, 2H), 7.57 (d, J=12 Hz, 2H), 3.36-3.18(m, 2H), 1.34(t, J=7.4 Hz, 3H)^[20]
4h	White solid	51	¹H NMR (400 MHz, DMSO-d₆)δ 8.00(d, J=8.4 Hz, 2H), 7.60(s, 2H), 7.42(d, J=7.2 Hz, 2H), 7.32(t, J=7.2 Hz, 2H), 7.42(d, J=7.2 Hz, 1H), 4.45(s, 2H)^[21]

Compd.

Apearance

Yield/%

¹H NMR

White solid

¹H NMR (400 MHz, DMSO-d₆)δ10.13(s, 1H), 9.43(s, 1H), 8.10(d, J=8.0 Hz, 2H), 7.95(s, 1H), 7.88(d, J=8.0 Hz, 2H)^[15]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ 14.09(s, 1H), 13.83(s, 1H), 8.12(d, J=8.4 Hz, 2H), 7.91(d, J=8.4 Hz, 2H)^[16]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ 8.19(d, J=8.8 Hz, 2H), 7.88(s, 2H), 3.21(d, J=1.6 Hz, 2H), 1.34(t, J=7.2 Hz, 3H)^[17]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ10.47(s, 1H), 9.37(s, 1H), 7.91(d, J=8.4 Hz, 2H), 7.70(s, 1H), 7.57(d, J=8.4 Hz, 2H)^[18]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ 13.93(s, 1H), 13.75(s, 1H), 7.92(d, J=8.4 Hz, 2H), 7.61(d, J=8.4 Hz, 2H)^[19]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ7.98(d, J=8 Hz, 2H), 7.57 (d, J=12 Hz, 2H), 3.36-3.18(m, 2H), 1.34(t, J=7.4 Hz, 3H)^[20]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ 8.00(d, J=8.4 Hz, 2H), 7.60(s, 2H), 7.42(d, J=7.2 Hz, 2H), 7.32(t, J=7.2 Hz, 2H), 7.42(d, J=7.2 Hz, 1H), 4.45(s, 2H)^[21]

Compd.	Apearance	Yield/%	¹H NMR
2b	White solid	43	¹H NMR (400 MHz, DMSO-d₆)δ10.13(s, 1H), 9.43(s, 1H), 8.10(d, J=8.0 Hz, 2H), 7.95(s, 1H), 7.88(d, J=8.0 Hz, 2H)^[15]
3b	White solid	53	¹H NMR (400 MHz, DMSO-d₆)δ 14.09(s, 1H), 13.83(s, 1H), 8.12(d, J=8.4 Hz, 2H), 7.91(d, J=8.4 Hz, 2H)^[16]
4b	White solid	47	¹H NMR (400 MHz, DMSO-d₆)δ 8.19(d, J=8.8 Hz, 2H), 7.88(s, 2H), 3.21(d, J=1.6 Hz, 2H), 1.34(t, J=7.2 Hz, 3H)^[17]
2c	White solid	56	¹H NMR (400 MHz, DMSO-d₆)δ10.47(s, 1H), 9.37(s, 1H), 7.91(d, J=8.4 Hz, 2H), 7.70(s, 1H), 7.57(d, J=8.4 Hz, 2H)^[18]
3c	White solid	60	¹H NMR (400 MHz, DMSO-d₆)δ 13.93(s, 1H), 13.75(s, 1H), 7.92(d, J=8.4 Hz, 2H), 7.61(d, J=8.4 Hz, 2H)^[19]
4c	White solid	59	¹H NMR (400 MHz, DMSO-d₆)δ7.98(d, J=8 Hz, 2H), 7.57 (d, J=12 Hz, 2H), 3.36-3.18(m, 2H), 1.34(t, J=7.4 Hz, 3H)^[20]
4h	White solid	51	¹H NMR (400 MHz, DMSO-d₆)δ 8.00(d, J=8.4 Hz, 2H), 7.60(s, 2H), 7.42(d, J=7.2 Hz, 2H), 7.32(t, J=7.2 Hz, 2H), 7.42(d, J=7.2 Hz, 1H), 4.45(s, 2H)^[21]

Compd.

Apearance

Yield/%

¹H NMR

White solid

¹H NMR (400 MHz, DMSO-d₆)δ10.13(s, 1H), 9.43(s, 1H), 8.10(d, J=8.0 Hz, 2H), 7.95(s, 1H), 7.88(d, J=8.0 Hz, 2H)^[15]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ 14.09(s, 1H), 13.83(s, 1H), 8.12(d, J=8.4 Hz, 2H), 7.91(d, J=8.4 Hz, 2H)^[16]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ 8.19(d, J=8.8 Hz, 2H), 7.88(s, 2H), 3.21(d, J=1.6 Hz, 2H), 1.34(t, J=7.2 Hz, 3H)^[17]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ10.47(s, 1H), 9.37(s, 1H), 7.91(d, J=8.4 Hz, 2H), 7.70(s, 1H), 7.57(d, J=8.4 Hz, 2H)^[18]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ 13.93(s, 1H), 13.75(s, 1H), 7.92(d, J=8.4 Hz, 2H), 7.61(d, J=8.4 Hz, 2H)^[19]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ7.98(d, J=8 Hz, 2H), 7.57 (d, J=12 Hz, 2H), 3.36-3.18(m, 2H), 1.34(t, J=7.4 Hz, 3H)^[20]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ 8.00(d, J=8.4 Hz, 2H), 7.60(s, 2H), 7.42(d, J=7.2 Hz, 2H), 7.32(t, J=7.2 Hz, 2H), 7.42(d, J=7.2 Hz, 1H), 4.45(s, 2H)^[21]

Compd.	Apearance	Yield/%	mp/℃	¹H NMR, ¹³C NMR and HR-MS
b	White solid	44	76.3-76.9	¹H NMR (400 MHz, CDCl₃)δ 8.29(d, J=8.0 Hz, 4H), 7.73(d, J=8.4 Hz, 2H), 7.68(d, J=7.2 Hz, 1H), 7.56(t, J=7.6 Hz, 2H), 3.42(q, J=7.2 Hz, 2H), 1.52(t, J=7.2 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 166.04, 162.07, 161.10, 133.73, 133.1 6, 132.09, 131.66, 130.89, 128.27, 127.38, 125.60, 125.56, 27.18, 13.96. HR-MS (ESI) C₁₈H₁₄F₃N₃OS [M-H]^- Calcd.: 378.088 8, Found: 378.089 0
c	White solid	44	79.2-80.1	¹H NMR (400 MHz, CDCl₃)δ 8.78(t, J₁=7.2 Hz, J₂=1.6 Hz, 2H), 8.12(t, J₁=1.6 Hz, J₂=6.8 Hz, 2H), 7.68(t, J₁=7.6 Hz, J₂=7.6 Hz, 1H), 7.56(t, J₁=8.0 Hz, J₂=7.6 Hz, 2H), 7.45(d, J=8.8 Hz, 2H), 3.44-3.38(m, 2H), 1.52(t, J₁=7.6 Hz, J₂=7.2 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 166.03, 161.82, 161.44, 136.31, 133.62, 131.66, 131.01, 128.88, 128.42, 128.28, 128.23, 27.15, 13.99. HR-MS (ESI) C₁₇H₁₄ClN₃OS [M-H]^- Calcd.: 344.062 4, Found: 344.062 7
d	White solid	34	114.7-115.6	¹H NMR (400 MHz, CDCl₃)δ 8.31(d, J=8.0 Hz, 2H), 8.21(d, J=3.6 Hz, 1H), 7.85(s, 1H), 7.76(d, J=8.0 Hz, 2H), 6.71(d, J=2.0 Hz, 1H), 3.39(q, J=7.2 Hz, 2H), 1.54(t, J=7.2 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 161.98, 161.42, 154.54, 149.10, 144.07, 133.07, 132.18, 127.35, 125.65, 125.61, 124.97, 112.80, 27.06, 13.93. HR-MS (ESI) C₁₆H₁₃F₃N₃O₂S [M-H]^- Calcd.: 368.068 1, Found: 368.068 5
e	White solid	40	127.0-127.5	¹H NMR (400 MHz, CDCl₃)δ 8.21(d, J=3.6 Hz, 1H), 8.15(d, J=8.8 Hz, 2H), 7.85(s, 1H), 7.48(d, J=8.4 Hz, 2H), 6.70(dd, J₁=3.6 Hz, J₂=1.6 Hz, 1H)3.36(q, J=7.2 Hz, 2H), 1.52(t, J=7.2 Hz, 3H).¹³C NMR (100 MHz, CDCl₃)δ 161.76, 161.74, 154.58, 148.98, 144.16, 136.42, 128.94, 128.38, 128.20, 124.89, 112.76, 27.03, 13.97. HR-MS (ESI) C₁₅H₁₂ClN₃O₂S [M-H]^- Calcd.: 334.041 7, Found: 334.041 5
f	White solid	37	94.9-96.5	¹H NMR (400 MHz, CDCl₃)δ 8.26(d, J=8.0 Hz, 2H), 7.71(d, J=8.0 Hz, 2H), 3.33(q, J=7.6 Hz, 2H), 3.16(q, J=7.6 Hz, 2H), 1.51(t, J=7.6 Hz, 3H), 1.31(t, J=7.6 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 173.04, 160.86, 159.87, 133.22, 132.03, 131.71, 127.25, 125.59, 125.55, 28.70, 26.84, 13.98, 7.88. HR-MS (ESI) C₁₄H₁₅F₃N₃OS [M-H]^-Calcd.: 330.088 8, Found: 330.089 2
g	White solid	41	116.1-116.7	¹H NMR (400 MHz, CDCl₃)δ 8.08(d, J=8.4 Hz, 2H), 7.43(d, J=8.8 Hz, 2H), 3.32(q, J=7.6 Hz, 2H), 3.14(q, J=7.6 Hz, 2H), 1.48(t, J=7.6 Hz, 3H), 1.29(t, J=7.6 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 173.04, 161.19, 159.62, 136.22, 128.87, 128.35, 138.29, 28.69, 26.80, 14.00, 7.91. HR-MS (ESI) C₁₃H₁₅ClN₃OS [M-H]^- Calcd.: 296.062 4, Found: 296.062 2
h	White solid	38	158.3-159.5	¹H NMR (400 MHz, CDCl₃)δ 8.25(d, J=7.2 Hz, 2H), 8.11(d, J=8.8 Hz, 2H), 7.65(t, J=7.6 Hz, 1H), 7.53(t, J=8.0 Hz, 4H), 7.43(d, J=8.4 Hz, 2H), 7.35(t, J=7.2 Hz, 2H), 7.29(d, J=7.2 Hz, 1H), 4.61(s, 2H). ¹³C NMR (100 MHz, CDCl₃)δ165.97, 161.40, 136.41, 136.34, 133.72, 131.69, 130.80, 130.61, 129.46, 128.97, 128.69, 128.43, 128.26, 128.24, 127.72, 37.30. HR-MS (ESI) C₂₂H₁₇ClN₃OS [M-H]^- Calcd.: 406.078 1, Found: 406.078 2
i	White solid	37	163.6-165.3	¹H NMR (400 MHz, CDCl₃)δ 8.18(d, J=3.2 Hz, 1H), 8.14(d, J=8.4 Hz, 2H), 7.82(s, 1H), 7.51(s, 1H), 7.49(q, J=3.2 Hz, 2H), 7.47(s, 1H), 7.34(t, J=7.2 Hz, 2H), 7.29(d, J=7.2 Hz, 1H), 6.68(m, 1H), 4.60(s, 2H). ¹³C NMR (100 MHz, CDCl₃)δ 161.72, 161.28, 154.52, 149.09, 144.02, 136.52, 136.27, 129.43, 129.03, 128.66, 128.39, 128.16, 127.70, 125.01, 112.79, 37.15. HR-MS (ESI) C₂₀H₁₄ClN₃O₂S [M-H]^- Calcd.: 396.057 3, Found: 396.057 0

Compd.

Apearance

Yield/%

mp/℃

¹H NMR, ¹³C NMR and HR-MS

White solid

76.3-76.9

¹H NMR (400 MHz, CDCl₃)δ 8.29(d, J=8.0 Hz, 4H), 7.73(d, J=8.4 Hz, 2H), 7.68(d, J=7.2 Hz, 1H), 7.56(t, J=7.6 Hz, 2H), 3.42(q, J=7.2 Hz, 2H), 1.52(t, J=7.2 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 166.04, 162.07, 161.10, 133.73, 133.1 6, 132.09, 131.66, 130.89, 128.27, 127.38, 125.60, 125.56, 27.18, 13.96. HR-MS (ESI) C₁₈H₁₄F₃N₃OS [M-H]^- Calcd.: 378.088 8, Found: 378.089 0

White solid

79.2-80.1

¹H NMR (400 MHz, CDCl₃)δ 8.78(t, J₁=7.2 Hz, J₂=1.6 Hz, 2H), 8.12(t, J₁=1.6 Hz, J₂=6.8 Hz, 2H), 7.68(t, J₁=7.6 Hz, J₂=7.6 Hz, 1H), 7.56(t, J₁=8.0 Hz, J₂=7.6 Hz, 2H), 7.45(d, J=8.8 Hz, 2H), 3.44-3.38(m, 2H), 1.52(t, J₁=7.6 Hz, J₂=7.2 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 166.03, 161.82, 161.44, 136.31, 133.62, 131.66, 131.01, 128.88, 128.42, 128.28, 128.23, 27.15, 13.99. HR-MS (ESI) C₁₇H₁₄ClN₃OS [M-H]^- Calcd.: 344.062 4, Found: 344.062 7

White solid

114.7-115.6

¹H NMR (400 MHz, CDCl₃)δ 8.31(d, J=8.0 Hz, 2H), 8.21(d, J=3.6 Hz, 1H), 7.85(s, 1H), 7.76(d, J=8.0 Hz, 2H), 6.71(d, J=2.0 Hz, 1H), 3.39(q, J=7.2 Hz, 2H), 1.54(t, J=7.2 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 161.98, 161.42, 154.54, 149.10, 144.07, 133.07, 132.18, 127.35, 125.65, 125.61, 124.97, 112.80, 27.06, 13.93. HR-MS (ESI) C₁₆H₁₃F₃N₃O₂S [M-H]^- Calcd.: 368.068 1, Found: 368.068 5

White solid

127.0-127.5

¹H NMR (400 MHz, CDCl₃)δ 8.21(d, J=3.6 Hz, 1H), 8.15(d, J=8.8 Hz, 2H), 7.85(s, 1H), 7.48(d, J=8.4 Hz, 2H), 6.70(dd, J₁=3.6 Hz, J₂=1.6 Hz, 1H)3.36(q, J=7.2 Hz, 2H), 1.52(t, J=7.2 Hz, 3H).¹³C NMR (100 MHz, CDCl₃)δ 161.76, 161.74, 154.58, 148.98, 144.16, 136.42, 128.94, 128.38, 128.20, 124.89, 112.76, 27.03, 13.97. HR-MS (ESI) C₁₅H₁₂ClN₃O₂S [M-H]^- Calcd.: 334.041 7, Found: 334.041 5

White solid

94.9-96.5

¹H NMR (400 MHz, CDCl₃)δ 8.26(d, J=8.0 Hz, 2H), 7.71(d, J=8.0 Hz, 2H), 3.33(q, J=7.6 Hz, 2H), 3.16(q, J=7.6 Hz, 2H), 1.51(t, J=7.6 Hz, 3H), 1.31(t, J=7.6 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 173.04, 160.86, 159.87, 133.22, 132.03, 131.71, 127.25, 125.59, 125.55, 28.70, 26.84, 13.98, 7.88. HR-MS (ESI) C₁₄H₁₅F₃N₃OS [M-H]^-Calcd.: 330.088 8, Found: 330.089 2

White solid

116.1-116.7

¹H NMR (400 MHz, CDCl₃)δ 8.08(d, J=8.4 Hz, 2H), 7.43(d, J=8.8 Hz, 2H), 3.32(q, J=7.6 Hz, 2H), 3.14(q, J=7.6 Hz, 2H), 1.48(t, J=7.6 Hz, 3H), 1.29(t, J=7.6 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 173.04, 161.19, 159.62, 136.22, 128.87, 128.35, 138.29, 28.69, 26.80, 14.00, 7.91. HR-MS (ESI) C₁₃H₁₅ClN₃OS [M-H]^- Calcd.: 296.062 4, Found: 296.062 2

White solid

158.3-159.5

¹H NMR (400 MHz, CDCl₃)δ 8.25(d, J=7.2 Hz, 2H), 8.11(d, J=8.8 Hz, 2H), 7.65(t, J=7.6 Hz, 1H), 7.53(t, J=8.0 Hz, 4H), 7.43(d, J=8.4 Hz, 2H), 7.35(t, J=7.2 Hz, 2H), 7.29(d, J=7.2 Hz, 1H), 4.61(s, 2H). ¹³C NMR (100 MHz, CDCl₃)δ165.97, 161.40, 136.41, 136.34, 133.72, 131.69, 130.80, 130.61, 129.46, 128.97, 128.69, 128.43, 128.26, 128.24, 127.72, 37.30. HR-MS (ESI) C₂₂H₁₇ClN₃OS [M-H]^- Calcd.: 406.078 1, Found: 406.078 2

White solid

163.6-165.3

¹H NMR (400 MHz, CDCl₃)δ 8.18(d, J=3.2 Hz, 1H), 8.14(d, J=8.4 Hz, 2H), 7.82(s, 1H), 7.51(s, 1H), 7.49(q, J=3.2 Hz, 2H), 7.47(s, 1H), 7.34(t, J=7.2 Hz, 2H), 7.29(d, J=7.2 Hz, 1H), 6.68(m, 1H), 4.60(s, 2H). ¹³C NMR (100 MHz, CDCl₃)δ 161.72, 161.28, 154.52, 149.09, 144.02, 136.52, 136.27, 129.43, 129.03, 128.66, 128.39, 128.16, 127.70, 125.01, 112.79, 37.15. HR-MS (ESI) C₂₀H₁₄ClN₃O₂S [M-H]^- Calcd.: 396.057 3, Found: 396.057 0

Compd.	Apearance	Yield/%	mp/℃	¹H NMR, ¹³C NMR and HR-MS
b	White solid	44	76.3-76.9	¹H NMR (400 MHz, CDCl₃)δ 8.29(d, J=8.0 Hz, 4H), 7.73(d, J=8.4 Hz, 2H), 7.68(d, J=7.2 Hz, 1H), 7.56(t, J=7.6 Hz, 2H), 3.42(q, J=7.2 Hz, 2H), 1.52(t, J=7.2 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 166.04, 162.07, 161.10, 133.73, 133.1 6, 132.09, 131.66, 130.89, 128.27, 127.38, 125.60, 125.56, 27.18, 13.96. HR-MS (ESI) C₁₈H₁₄F₃N₃OS [M-H]^- Calcd.: 378.088 8, Found: 378.089 0
c	White solid	44	79.2-80.1	¹H NMR (400 MHz, CDCl₃)δ 8.78(t, J₁=7.2 Hz, J₂=1.6 Hz, 2H), 8.12(t, J₁=1.6 Hz, J₂=6.8 Hz, 2H), 7.68(t, J₁=7.6 Hz, J₂=7.6 Hz, 1H), 7.56(t, J₁=8.0 Hz, J₂=7.6 Hz, 2H), 7.45(d, J=8.8 Hz, 2H), 3.44-3.38(m, 2H), 1.52(t, J₁=7.6 Hz, J₂=7.2 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 166.03, 161.82, 161.44, 136.31, 133.62, 131.66, 131.01, 128.88, 128.42, 128.28, 128.23, 27.15, 13.99. HR-MS (ESI) C₁₇H₁₄ClN₃OS [M-H]^- Calcd.: 344.062 4, Found: 344.062 7
d	White solid	34	114.7-115.6	¹H NMR (400 MHz, CDCl₃)δ 8.31(d, J=8.0 Hz, 2H), 8.21(d, J=3.6 Hz, 1H), 7.85(s, 1H), 7.76(d, J=8.0 Hz, 2H), 6.71(d, J=2.0 Hz, 1H), 3.39(q, J=7.2 Hz, 2H), 1.54(t, J=7.2 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 161.98, 161.42, 154.54, 149.10, 144.07, 133.07, 132.18, 127.35, 125.65, 125.61, 124.97, 112.80, 27.06, 13.93. HR-MS (ESI) C₁₆H₁₃F₃N₃O₂S [M-H]^- Calcd.: 368.068 1, Found: 368.068 5
e	White solid	40	127.0-127.5	¹H NMR (400 MHz, CDCl₃)δ 8.21(d, J=3.6 Hz, 1H), 8.15(d, J=8.8 Hz, 2H), 7.85(s, 1H), 7.48(d, J=8.4 Hz, 2H), 6.70(dd, J₁=3.6 Hz, J₂=1.6 Hz, 1H)3.36(q, J=7.2 Hz, 2H), 1.52(t, J=7.2 Hz, 3H).¹³C NMR (100 MHz, CDCl₃)δ 161.76, 161.74, 154.58, 148.98, 144.16, 136.42, 128.94, 128.38, 128.20, 124.89, 112.76, 27.03, 13.97. HR-MS (ESI) C₁₅H₁₂ClN₃O₂S [M-H]^- Calcd.: 334.041 7, Found: 334.041 5
f	White solid	37	94.9-96.5	¹H NMR (400 MHz, CDCl₃)δ 8.26(d, J=8.0 Hz, 2H), 7.71(d, J=8.0 Hz, 2H), 3.33(q, J=7.6 Hz, 2H), 3.16(q, J=7.6 Hz, 2H), 1.51(t, J=7.6 Hz, 3H), 1.31(t, J=7.6 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 173.04, 160.86, 159.87, 133.22, 132.03, 131.71, 127.25, 125.59, 125.55, 28.70, 26.84, 13.98, 7.88. HR-MS (ESI) C₁₄H₁₅F₃N₃OS [M-H]^-Calcd.: 330.088 8, Found: 330.089 2
g	White solid	41	116.1-116.7	¹H NMR (400 MHz, CDCl₃)δ 8.08(d, J=8.4 Hz, 2H), 7.43(d, J=8.8 Hz, 2H), 3.32(q, J=7.6 Hz, 2H), 3.14(q, J=7.6 Hz, 2H), 1.48(t, J=7.6 Hz, 3H), 1.29(t, J=7.6 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 173.04, 161.19, 159.62, 136.22, 128.87, 128.35, 138.29, 28.69, 26.80, 14.00, 7.91. HR-MS (ESI) C₁₃H₁₅ClN₃OS [M-H]^- Calcd.: 296.062 4, Found: 296.062 2
h	White solid	38	158.3-159.5	¹H NMR (400 MHz, CDCl₃)δ 8.25(d, J=7.2 Hz, 2H), 8.11(d, J=8.8 Hz, 2H), 7.65(t, J=7.6 Hz, 1H), 7.53(t, J=8.0 Hz, 4H), 7.43(d, J=8.4 Hz, 2H), 7.35(t, J=7.2 Hz, 2H), 7.29(d, J=7.2 Hz, 1H), 4.61(s, 2H). ¹³C NMR (100 MHz, CDCl₃)δ165.97, 161.40, 136.41, 136.34, 133.72, 131.69, 130.80, 130.61, 129.46, 128.97, 128.69, 128.43, 128.26, 128.24, 127.72, 37.30. HR-MS (ESI) C₂₂H₁₇ClN₃OS [M-H]^- Calcd.: 406.078 1, Found: 406.078 2
i	White solid	37	163.6-165.3	¹H NMR (400 MHz, CDCl₃)δ 8.18(d, J=3.2 Hz, 1H), 8.14(d, J=8.4 Hz, 2H), 7.82(s, 1H), 7.51(s, 1H), 7.49(q, J=3.2 Hz, 2H), 7.47(s, 1H), 7.34(t, J=7.2 Hz, 2H), 7.29(d, J=7.2 Hz, 1H), 6.68(m, 1H), 4.60(s, 2H). ¹³C NMR (100 MHz, CDCl₃)δ 161.72, 161.28, 154.52, 149.09, 144.02, 136.52, 136.27, 129.43, 129.03, 128.66, 128.39, 128.16, 127.70, 125.01, 112.79, 37.15. HR-MS (ESI) C₂₀H₁₄ClN₃O₂S [M-H]^- Calcd.: 396.057 3, Found: 396.057 0

Compd.

Apearance

Yield/%

mp/℃

¹H NMR, ¹³C NMR and HR-MS

White solid

76.3-76.9

White solid

79.2-80.1

White solid

114.7-115.6

White solid

127.0-127.5

White solid

94.9-96.5

White solid

116.1-116.7

White solid

158.3-159.5

White solid

163.6-165.3


a	Cl		CH₃	58.87	5.84±0.01
b	CF₃	CH₂CH₃	75.00	2.31±0.15
c	Cl	CH₂CH₃	74.42	3.18±0.11
d	CF₃	CH₂CH₃	81.98	1.50±0.04
e	Cl	CH₂CH₃	75.57	2.85±0.38
f	CF₃	CH₂CH₃	74.81	2.94±0.37
g	Cl	CH₂CH₃	65.76	4.68±0.12
h	Cl	Bn	30.01	> 10
i	Cl	Bn	37.89	> 10
Brequinar	-	-	-	-	0.01±0.00

Compd.

R¹

R²

R³

Inhibition rate (10 μmol·L^-1)/%

IC₅₀/μmol·L^-1a

CH₃

58.87

5.84±0.01

CF₃

CH₂CH₃

75.00

2.31±0.15

CH₂CH₃

74.42

3.18±0.11

CF₃

CH₂CH₃

81.98

1.50±0.04

CH₂CH₃

75.57

2.85±0.38

CF₃

CH₂CH₃

74.81

2.94±0.37

CH₂CH₃

65.76

4.68±0.12

30.01

> 10

37.89

> 10

Brequinar

0.01±0.00


a	Cl		CH₃	58.87	5.84±0.01
b	CF₃	CH₂CH₃	75.00	2.31±0.15
c	Cl	CH₂CH₃	74.42	3.18±0.11
d	CF₃	CH₂CH₃	81.98	1.50±0.04
e	Cl	CH₂CH₃	75.57	2.85±0.38
f	CF₃	CH₂CH₃	74.81	2.94±0.37
g	Cl	CH₂CH₃	65.76	4.68±0.12
h	Cl	Bn	30.01	> 10
i	Cl	Bn	37.89	> 10
Brequinar	-	-	-	-	0.01±0.00

Compd.

R¹

R²

R³

Inhibition rate (10 μmol·L^-1)/%

IC₅₀/μmol·L^-1a

CH₃

58.87

5.84±0.01

CF₃

CH₂CH₃

75.00

2.31±0.15

CH₂CH₃

74.42

3.18±0.11

CF₃

CH₂CH₃

81.98

1.50±0.04

CH₂CH₃

75.57

2.85±0.38

CF₃

CH₂CH₃

74.81

2.94±0.37

CH₂CH₃

65.76

4.68±0.12

30.01

> 10

37.89

> 10

Brequinar

0.01±0.00

二氢乳清酸脱氢酶抑制剂的设计、合成和构效关系研究

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宫荥恢 , 刘莉 , 齐甜甜 , 李洪林 , 朱丽丽 ^* , 赵振江 ^*

药学学报 | 药物化学 2017,52(2): 264-270

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药学学报 | 药物化学 2017, 52(2): 264-270

二氢乳清酸脱氢酶抑制剂的设计、合成和构效关系研究

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宫荥恢, 刘莉, 齐甜甜, 李洪林, 朱丽丽^*, 赵振江^*

作者信息

华东理工大学药学院, 上海市新药设计重点实验室, 上海 200237

通讯作者:

* 朱丽丽, Tel:86-21-64253962, E-mail:zhulfl@ecust.edu.cn;

赵振江, E-mail:zhjzhao@ecust.edu.cn

Design, synthesis and structure-activity relationship studies of human dihydroorotate dehydrogenase inhibitors

Ying-hui GONG, Li LIU, Tian-tian QI, Hong-lin LI, Li-li ZHU^*, Zhen-jiang ZHAO^*

Affiliations

Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China

出版时间: 2017-02-12 doi: 10.16438/j.0513-4870.2016-1120

文章导航

摘要

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本文将实验室前期发现的1-丁酰基-3-（4-氯苯基）-5-甲硫基-1H-1，2，4-三氮唑作为人类二氢乳清酸脱氢酶（HsDHODH）抑制剂设计的苗头化合物，为开展结构优化，首先培养并解析了苗头化合物与HsDHODH的复合物晶体结构；在此基础上，设计合成了一系列基于苗头结构新颖的HsDHODH抑制剂。活性结果表明，新合成的目标化合物对HsDHODH具有较好的抑制活性，其中1-呋喃甲酰基-3-（4-三氟甲基苯基）-5-乙硫基-1H-1，2，4-三氮唑的IC₅₀值达到1.50μmol·L^-1。结合生物学活性测试及复合物晶体结构对该系列化合物的构效关系进行了分析与总结，为1H-1，2，4-三氮唑衍生物HsDHODH抑制剂的进一步优化提供了借鉴经验。

关键词

人类二氢乳清酸脱氢酶抑制剂 / 三氮唑衍生物 / 自身免疫性疾病 / 复合物共晶结构 / 构效关系

Abstract

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In this study, 1-(3-(4-chlorophenyl)-5-methylthio-1H-1, 2, 4-triazol-1-yl)-butan-1-one discovered previously in our lab was selected as a inhibitor of human dihydroorotate dehydrogenase (HsDHODH) for structural optimization. The co-crystal of HsDHODH with the hit was obtained and analyzed for guiding the subsequent structural optimization. As a result, a series of novel triazole derivatives were designed and synthesized as potent HsDHODH inhibitors. Among them, compound (3-(4-chlorophenyl)-5-ethylthio-1H-1, 2, 4-triazol-1-yl)-furan-2-yl-methanone displayed high potency in the inhibition of HsDHODH with an IC₅₀ value of 1.50 μmol·L^-1. Meanwhile, the structure-activity relationships were analyzed based on the biological data and the co-crystal structure. These results provide a valuable reference for optimization of 1H-1, 2, 4-triazole derivatives as HsDHODH inhibitors in the future.

Key words

human dihydroorotate dehydrogenase inhibitor / triazole derivative / autoimmune disease / complex co-crystal structure / structure-activity relationship

引用本文

宫荥恢, 刘莉, 齐甜甜, 李洪林, 朱丽丽, 赵振江. 二氢乳清酸脱氢酶抑制剂的设计、合成和构效关系研究. 药学学报, 2017 , 52 (2) : 264 -270 . DOI: 10.16438/j.0513-4870.2016-1120

Ying-hui GONG, Li LIU, Tian-tian QI, Hong-lin LI, Li-li ZHU, Zhen-jiang ZHAO. Design, synthesis and structure-activity relationship studies of human dihydroorotate dehydrogenase inhibitors[J]. Acta Pharmaceutica Sinica, 2017 , 52 (2) : 264 -270 . DOI: 10.16438/j.0513-4870.2016-1120

正文

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二氢乳清酸脱氢酶(dihydroorotate dehydrogenase, DHODH) 是一种催化嘧啶核苷酸从头合成第4步反应的黄素依赖性线粒体酶^[1]。人类二氢乳清酸脱氢酶(HsDHODH) 抑制剂可以阻断嘧啶核苷酸生物从头合成途径, 阻止细胞进入DNA复制的S期, 进而减少快速增殖的淋巴细胞及肿瘤细胞的增殖。因此, HsDHODH可作为癌症和类风湿性关节炎、红斑狼疮、牛皮癣及多发性硬化症等自身免疫性疾病药物的靶点^[2]。

DHODH分为两个家族: Ⅰ型和Ⅱ型。HsDHODH属于家族Ⅱ型酶, 以辅酶Q作为电子受体^{[3, 4]}。在DHODH参与的催化反应过程中, 首先二氢乳清酸(DHO) 被氧化为乳清酸(ORO), 同时黄素单核苷酸(FMN) 被还原为还原型黄素单核苷酸(FMNH₂), 其后在辅酶Q的作用下FMNH₂被重新氧化为FMN, 参与到下一轮的催化循环反应中^[5]。

布喹那(化合物1, 图 1) 和来氟米特(化合物2, 图 1) 是两种比较典型的DHODH抑制剂^{[6, 7]}。来氟米特是唯一一个已上市的用于治疗类风湿性关节炎的HsDHODH抑制剂, 然而来氟米特的长期使用会引发较大的不良反应如腹泻、高血压、皮疹, 并且会导致肝功能损伤严重^{[8, 9]}。布喹那联合顺铂或是环孢霉素A给药时, 会引起白细胞和血小板减少以及黏膜炎等不良反应^{[10, 11]}。因此, 寻找高效新型、不良反应小、安全性高的DHODH抑制剂具有很高的研究价值。首先, 在布喹那和来氟米特基础上对其结构进行了优化尝试。化合物4(FK778) 进一步延长了A771726烯醇上的疏水基团, 该化合物在2004年曾作为抑制肾移植排异反应的药物进行临床试验^{[12, 13]}。2012年, 本课题组采用分子对接程序Glide和结合自由能计算法Prime/MM-GBSA联用的策略, 获得噻唑衍生物HsDHODH抑制剂先导物, 经过优化得到化合物5(IC₅₀=18 nmol·L^-1), 同时对于CIA关节炎模型大鼠的关节肿胀有显著缓解, 且组织病理学切片镜检显示关节组织病变有明显改善^{[14, 15]}。

为寻找新型HsDHODH抑制剂, 本文以课题组前期发现的带有1, 2, 4-三氮唑母核的1-丁酰基-3-(4-氯苯基)-5-甲硫基-1H-1, 2, 4-三氮唑作为苗头化合物^[14], 其抑制活性IC₅₀值为5.84 μmol·L^-1。为开展结构优化, 培养和分析了苗头化合物与DHODH的复合物晶体结构, 并制订了结构改造方案。以取代苯甲酸和氨基硫脲为原料, 设计合成了一系列结构新颖且具有较高抑制活性的基于三氮唑骨架的潜在人类二氢乳清酸脱氢酶抑制剂, 产物结构经¹H NMR、¹³C NMR和HR-MS确认, 并对其构效关系进行深入研究, 目标化合物的合成路线如下所示(合成路线1)。

结果与讨论

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1 苗头化合物a与HsDHODH共晶培养与解析

为进一步阐述苗头化合物a与HsDHODH的结合模式, 提高后续结构优化的有效性, 培养了苗头化合物a与HsDHODH的复合物晶体(表 1)。

苗头化合物a与HsDHODH复合物的晶体结构如图 2所示。苯环上的氯原子位于酶口袋深处由氨基酸残基Val134和Val143形成的一个小的疏水空腔中而产生疏水相互作用。同时, 1, 2, 4-三氮唑环和甲硫基处于辅酶Q结合口袋入口处较大的疏水空腔中, 整个分子主要通过与酶口袋活性位点处的氨基酸产生疏水相互作用而保持较高的抑制活性。

2 苗头化合物a结构优化策略

基于对苗头化合物与蛋白酶复合物晶体结构解析, 将苗头化合物展开以下几个方面的优化, 如图 3所示:为增强疏水性相互作用, 探寻与酶口袋内部由氨基酸残基Val134和Val143组成的小的疏水空腔相匹配的苯环取代基, 本研究将在苗头化合物中R¹处引入疏水基团; 为通过增强抑制剂与Leu42、Ala59、Phe62、Leu68、Phe98、Met111和Leu359等残基的疏水作用和范德华力, 本研究将在苗头化合物中R²处引入一些疏水性基团; 针对苗头化合物中R³处疏水基团的优化, 希望找到与口袋疏水部分相匹配的疏水取代基, 进而增强化合物的抑制活性。

3 中间体和目标化合物的表征

中间体的物理性质及¹H NMR数据见表 2^[15-21], 目标化合物表征见表 3。

4 目标化合物构效关系讨论

合成的目标化合物HsDHODH活性测试结果见表 4。比较化合物b与c、d与e、f与g的活性可以看出, 三氟甲基的引入比苗头化合物中的氯原子可较好地提高抑制活性, 化合物d比化合物e抑制活性提高一倍。原因可能是三氟甲基的体积较氯原子略大, 疏水相互作用的加强导致抑制活性的提高。在R²引入呋喃甲酰基时, 活性得到改善, 其中化合物d的IC₅₀值可达1.50 μmol·L^-1, 明显优于苗头化合物的抑制活性。比较化合物h和i, 发现活性很差, 而化合物c和e表现出高活性, 这说明乙硫基的引入, 可更好地与酶口袋疏水部分相匹配, 苄硫基的空间体积过大或空间取向冲突, 导致与周边氨基酸残基发生碰撞而活性变差。

5 结论

本文基于苗头化合物与HsDHODH共晶结构培养和解析, 优化设计合成了一系列结构新颖的HsDHODH抑制剂, 并分析该系列化合物的构效关系。在苯环对位引入三氟甲基较氯原子可更好地占据酶口袋深处由氨基酸残基Val134和Val143组成的疏水凹槽, 有利于增强抑制活性; 疏水端三氮唑环R²处引入芳香环(如苯甲酰基、呋喃甲酰基) 较烷基更有利于增强与酶口袋入口处氨基酸残基Leu42、Ala59、Phe62、Leu68、Phe98、Met111和Leu359的π-π堆积疏水相互作用而提高化合物抑制活性, 化合物d的抑制活性达到1.50 μmol·L^-1, 明显优于苗头化合物的抑制活性; 三氮唑环R³处乙硫基的引入, 与苗头化合物中的甲硫基比较可加强与酶口袋周围氨基酸残基的疏水相互作用。值得注意的是, 在三氮唑环的氮原子上引入较大体积芳香基团的同时在硫原子上引入较小烷基链时, 由Leu46和Thr360包围的酶口袋处不足以容纳过大的基团, 化合物在酶口袋内部可发生近180度的空间构型旋转, 较大体积的芳香基团占据酶口袋入口处疏水空腔, 进而提高抑制活性。此结论对三氮唑环类HsDHODH抑制剂的进一步优化提供了参考, 有助于发现更高活性的HsDHODH抑制剂。

实验部分

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1 合成部分

核磁共振仪(瑞士, Bruker AV-400);显微熔点仪(上海精密科学仪器有限公司, SGW X-4);质谱仪(惠普, HP5989A); 柱色谱硅胶(200~300目, 青岛海洋化工有限公司); 药品与试剂均为分析纯。

目标化合物的合成(以目标化合物c为例):

对氯苯甲酰氯(1c):称量对氯苯甲酸(500 mg, 3.20 mmol) 于25 mL烧瓶中, 加入二氯亚砜(1 mL) 及一滴DMF。加热至回流, 反应过夜。过量的二氯亚砜减压蒸馏后, 制备的酰氯直接用于下步反应。

4-氯苯甲酰氨基硫脲(2c):称量氨基硫脲(230mg, 2.53 mmol) 于25 mL烧瓶中, 加入吡啶10 mL和中间体对氯苯甲酰氯(400 mg, 2.30 mmol), 加热至50 ℃, 反应12 h。将吡啶旋干, 剩余的固体, 加入水(10 mL) 搅拌, 过滤后得到白色的固体。得到的固体经过乙酸乙酯重结晶, 得到白色晶体。

3-(4-氯苯基)-1H-1, 2, 4-三氮唑-5(4H)-硫酮(3c):称量中间体4-氯苯甲酰氨基硫脲(200 mg, 0.87 mmol) 于25 mL烧瓶中, 加入1 mol·L^-1 NaHCO₃ (10mL), 加热至回流, 反应24 h, 用稀HCl中和至中性, 之后用乙酸乙酯萃取, 有机相加入无水硫酸钠干燥, 浓缩的有机相用乙酸乙酯和石油醚重结晶, 得到白色晶体。

3-(4-氯苯基)-5-乙硫基-1H-1, 2, 4-三氮唑(4c):称量中间体3-(4-氯苯基)-1H-1, 2, 4-三氮唑-5(4H)-硫酮(600 mg, 2.84 mmol) 于25 mL烧瓶中, 加入1 mol·L^-1 NaOH (6 mL), 量取碘乙烷(230 μL, 2.84mmol) 和乙醇(1 mL) 加入烧瓶中, 室温反应24 h。加入水用乙酸乙酯萃取, 收集有机相, 经过无水硫酸钠干燥后, 浓缩有机相, 经柱色谱纯化, 得到白色固体。

3-(4-氯苯基)-5-乙硫基-1H-1, 2, 4-三氮唑-1-基) 苯甲酮(目标化合物c):称量化合物3-(4-氯苯基)-5-乙硫基-1H-1, 2, 4-三氮唑(50.0 mg, 0.21 mmol) 和三乙烯二胺(70.7 mg, 0.63 mmol) 溶于无水DMF (1 mL) 中, 苯甲酰氯(88.6 mg, 0.63mmol) 溶于无水DMF (1 mL) 后加入上述溶液中, 氩气保护下, 在室温反应16 h。将溶液旋干后, 用二氯甲烷萃取, 用无水硫酸钠干燥。浓缩有机相, 得到油状物粗品, 用甲醇重结晶, 得到白色针状固体。

2 化合物的生物活性测试及苗头化合物-HsDHODH的复合物晶体培养

收起

2.1 化合物的生物活性测试

HsDHODH的蛋白表达和纯化方法参考文献所述^{[22, 23]}。HsDHODH的抑制剂筛选采用DCIP的方法^{[22, 24]}。将纯化后的HsDHODH蛋白用测活溶液(50 mmol·L^-1 HEPES pH 8.0, 150 mmol·L^-1 KCl, 0.1%Triton X-100) 稀释至10nmol·L^-1, 加入辅酶Q及DCIP至其终浓度分别为100及120 μmol·L^-1。混匀后用排枪以每孔199 μL加入到96孔板中, 之后置于室温下孵育5 min, 然后每孔加入1 μL底物DHO至其终浓度为500 μmol·L^-1。运用酶标仪读取600 nm处吸光值, 一共读取6 min, 每30 s读取一次。计算酶促反应初速度V₀, 抑制剂活性测试则是在上述反应体系中加入不同浓度的抑制剂, 计算酶促反应初速度V_i, 通过公式(1-V_i/V₀)×100%计算出化合物的抑制率。用软件Origin8.0计算化合物的IC₅₀值。该实验以brequinar为阳性对照, 每次实验至少设置3个平行。

2.2 HsDHODH与苗头化合物的共晶培养

苗头化合物和HsDHODH的复合晶体由悬滴蒸汽扩散法于20 ℃培养^[22]。将纯化后的HsDHODH蛋白浓缩至20 mg·mL^-1, 加入底物DHO及苗头化合物至终浓度均为2 mmol·L^-1, 然后置于冰上孵育2 h。沉淀剂溶液包含0.1mol·L^-1醋酸pH 4.8、40 mmol·L^-1 C11DAO、20mmol·L^-1 DDAO以及1.6~1.81 mol·L^-1硫酸铵。一般情况下在3天内晶体即以黄色小方块的形式显现, 在3周内晶体可达到0.2 mm × 0.2 mm × 0.2 mm的大小。

2.3 HsDHODH与苗头化合物的晶体衍射数据及结构解析

X射线晶体衍射数据是在上海同步辐射光源(SSRF) 生物大分子线站(BL17U1) 收集^[25], 温度为100 K。收集的衍射数据运用MOSFLM软件进行初步处理^[26], 用ccp4软件中的scale程序进行归并^[27], 接着以1D3G为模板(去掉所有的水分子和配体) 采用分子置换法进行解析, 用Coot^[24]和Refmac5^[28]进行交替精修。晶体衍射数据及结构解析见表 1。分子图形用Pymol (DeLano, 2002) 软件作图。

基金

收起

国家自然科学基金资助项目(21372078)
上海市科技支撑资助项目(14431902400)

参考文献

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文献

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参考文献引证文献

排序方式：

[1]

Das P, Deng X, Zhang L, et al. SAR-based optimization of a 4-quinoline carboxylic acidanalogue with potent antiviral activity[J]. ACS Med Chem Lett, 2013, 4:517-521.

[2]

Vincenti F, Cohen SD, Appel G. Novel B cell therapeutic targets in transplantation and immune-mediated glomerular diseases[J]. Clin J Am Soc Nephrol, 2010, 5:142-151.

[3]

Hansen M, Le Nours J, Johansson EVA, et al. Inhibitor binding in a class 2 dihydroorotate dehydrogenase causes variations in the membrane-associated N-terminal domain[J]. Protein Sci, 2004, 13:1031-1042.

[4]

Palfey BA, Björnberg O, Jensen KF. Insight into the chemistry of flavin reduction and oxidation in Escherichia coli dihydroorotate dehydrogenase obtained by rapid reaction studies[J]. Biochemistry, 2001, 40:4381-4390.

[5]

Malmquist NA, Gujjar R, Rathod PK, et al. Analysis of flavin oxidation and electron-transfer inhibition in Plasmodium falciparum dihydroorotate dehydrogenase[J]. Biochemistry, 2008, 47:2466-2475.

[6]

Gummert JF, Ikonen T, Morris RE. Newer immunosuppressive drugs a review[J]. J Am Soc Nephrol, 1999, 10:1366-1380.

[7]

Williamson RA, Yea CM, Robson PA, et al. Dihydroorotate dehydrogenase is a high affinity binding protein for A771726 and mediator of a range of biological effects of the immunomodulatory compound[J]. J Biol Chem, 1995, 270:22467-22472.

[8]

Herrmann ML, Schleyerbach R, Kirschbaum BJ. Leflunomide:an immunomodulatory drug for the treatment of rheumatoid arthritis and other autoimmune diseases[J]. Immunopharmacology, 2000, 47:273-289.

[9]

Alcorn N, Saunders S, Madhok R. Benefit-risk assessment of leflunomide[J]. Drug Safety, 2009, 32:1123-1134.

[10]

Gordon MS, Margolin K, Talpaz M, et al. Phase I safety and pharmacokinetic study of recombinant human anti-vascular endothelial growth factor in patients with advanced cancer[J]. J Clin Oncol, 2001, 19:843-850.

[11]

Pally C, Smith D, Jaffee B, et al. Side effects of brequinar and brequinar analogues, in combination with cyclosporine, in the rat[J]. Toxicology, 1998, 127:207-222.

[12]

Cullell-Young M, Castañer RM, Leeson PA. FK-778. Treatment of transplant rejection, dihydroorotate dehydrogenase inhibitor[J]. Drugs Future, 2002, 27:733-739.

[13]

Vanrenterghem Y, Van Hooff JP, Klinger M, et al. The effects of FK778 in combination with tacrolimus and steroids:a phase Ⅱ multicenter study in renal transplant patients[J]. Transplantation, 2004, 78:9-14.

[14]

Diao Y, Lu W, Jin H, et al. Discovery of diverse human dihydroorotate dehydrogenase inhibitors as immunosuppressive agents by structure-based virtual screening[J]. J Med Chem, 2012, 55:8341-8349.

[15]

Sharpe PL. Herbicidal triazoles:US, 033285[P]. 2016-03-03.

[16]

Cui J, Jin J, Chaudhary AS, et al. Design, synthesis and evaluation of triazole-pyrimidine analogues as seca inhibitors[J]. ChemMedChem, 2015, 11:43-56.

[17]

Sarkar D, Joshi RA, Likhite AP, et al. 1, 2, 4-Triazole, 1, 3, 4-oxadiazole, and 1, 3, 4-thiadiazole derivatives and their antimycobacterial activity:WO, 108249[P]. 2016-07-07.

[18]

Plumitallo A, Cardia MC, Distinto S, et al. Synthesis and anti-microbial activity evaluation of some new 1-benzoylisothiosemicarbazides[J]. Il Farmaco, 2004, 59:945-952.

[19]

Guda DR, Wang T, Cho HM, et al. Trimethylsilyl isothiocyanate (TMSNCS):an efficient reagent for the one-pot synthesis of mercapto-1, 2, 4-triazoles[J]. Tetrahedron Lett, 2012, 53:5238-5242.

[20]

Tozkoparan B, Küpeli E, Yeşilada E, et al. Preparation of 5-aryl-3-alkylthio-l, 2, 4-triazoles and corresponding sulfones with antiinflammatory-analgesic activity[J]. Bioorg Med Chem, 2007, 15:1808-1814.

[21]

Britsun VN, Esipenko AN, Lozinskii MO. Reaction of 2-aryl-5-R-5, 6-dihydro-7H-[1, 2, 4]-triazolo[5, 1-b] [1, 3] thiazin-7-ones with aryl bromomethyl ketones and benzyl bromide[J]. Chem Heterocycl Compd, 2005, 41:782-786.

[22]

Zhu J, Han L, Diao Y, et al. Design, synthesis, X-ray crystallographic analysis, and biological evaluation of thiazole derivatives as potent and selective inhibitors of human dihydroorotate dehydrogenase[J]. J Med Chem, 2015, 58:1123-1139.

[23]

Liu S, Neidhardt EA, Grossman TH, et al. Structures of human dihydroorotate dehydrogenase in complex with antiproliferative agents[J]. Structure, 2000, 8:25-33.

[24]

Li S, Luan G, Ren X, et al. Rational design of benzylidenehydrazinyl-substituted thiazole derivatives as potent inhibitors of human dihydroorotate dehydrogenase with in vivo antiarthritic activity[J]. Sci Rep, 2015, 5:14836.

[25]

Wang QS, Yu F, Huang S, et al. The macromolecular crystallography beamline of SSRF[J]. Nucl Sci Tech, 2015, 26:12-17.

[26]

Powell HR. The Rossmann Fourier autoindexing algorithm in MOSFLM[J]. Acta Crystallogr D Biol Crystallogr, 1999, 55(Pt 10):1690-1695.

[27]

Collaborative Computational Project, Number 4. The CCP4 suite:programs for protein crystallography[J]. Acta Crystallogr D Biol Crystallogr, 1994, 50(Pt 5):760-763.

[28]

Murshudov GN, Skubák P, Lebedev AA, et al. REFMAC5 for the refinement of macromolecular crystal structures[J]. Acta Crystallogr D Biol Crystallogr, 2011, 67(Pt 4):355-367.

2017年第52卷第2期

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doi: 10.16438/j.0513-4870.2016-1120

接收时间：2016-11-22
首发时间：2026-01-14
出版时间：2017-02-12

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收稿日期：2016-11-22
修回日期：2016-12-22

基金

国家自然科学基金资助项目(21372078)

上海市科技支撑资助项目(14431902400)

作者信息

华东理工大学药学院, 上海市新药设计重点实验室, 上海 200237

通讯作者:

* 朱丽丽, Tel:86-21-64253962, E-mail:zhulfl@ecust.edu.cn;

赵振江, E-mail:zhjzhao@ecust.edu.cn

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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Parameter	Data
Wavelengths/Å	0.97852
Space group	P 3₂ 2 1
Cell dimensions/Å	90.870/90.870/122.820
Resolution/Å	2.90
Number of reflections	12 608
Redundancy	7.2 (7.4)
Completeness/%	94.7 (95.9)
R_merge/%^a	28.3 (53.0)
Ⅰ/σ (Ⅰ)^b	6.6 (4.0)
R/R_free/%^c	15.77 (21.96)
Bonds/Å	0.010
Angles (deg.)	1.236

Parameter

Data

Wavelengths/Å

0.97852

Space group

P 3₂ 2 1

Cell dimensions/Å

90.870/90.870/122.820

Resolution/Å

2.90

Number of reflections

12 608

Redundancy

7.2 (7.4)

Completeness/%

94.7 (95.9)

R_merge/%^a

28.3 (53.0)

Ⅰ/σ (Ⅰ)^b

6.6 (4.0)

R/R_free/%^c

15.77 (21.96)

Bonds/Å

0.010

Angles (deg.)

1.236

Compd.	Apearance	Yield/%	¹H NMR
2b	White solid	43	¹H NMR (400 MHz, DMSO-d₆)δ10.13(s, 1H), 9.43(s, 1H), 8.10(d, J=8.0 Hz, 2H), 7.95(s, 1H), 7.88(d, J=8.0 Hz, 2H)^[15]
3b	White solid	53	¹H NMR (400 MHz, DMSO-d₆)δ 14.09(s, 1H), 13.83(s, 1H), 8.12(d, J=8.4 Hz, 2H), 7.91(d, J=8.4 Hz, 2H)^[16]
4b	White solid	47	¹H NMR (400 MHz, DMSO-d₆)δ 8.19(d, J=8.8 Hz, 2H), 7.88(s, 2H), 3.21(d, J=1.6 Hz, 2H), 1.34(t, J=7.2 Hz, 3H)^[17]
2c	White solid	56	¹H NMR (400 MHz, DMSO-d₆)δ10.47(s, 1H), 9.37(s, 1H), 7.91(d, J=8.4 Hz, 2H), 7.70(s, 1H), 7.57(d, J=8.4 Hz, 2H)^[18]
3c	White solid	60	¹H NMR (400 MHz, DMSO-d₆)δ 13.93(s, 1H), 13.75(s, 1H), 7.92(d, J=8.4 Hz, 2H), 7.61(d, J=8.4 Hz, 2H)^[19]
4c	White solid	59	¹H NMR (400 MHz, DMSO-d₆)δ7.98(d, J=8 Hz, 2H), 7.57 (d, J=12 Hz, 2H), 3.36-3.18(m, 2H), 1.34(t, J=7.4 Hz, 3H)^[20]
4h	White solid	51	¹H NMR (400 MHz, DMSO-d₆)δ 8.00(d, J=8.4 Hz, 2H), 7.60(s, 2H), 7.42(d, J=7.2 Hz, 2H), 7.32(t, J=7.2 Hz, 2H), 7.42(d, J=7.2 Hz, 1H), 4.45(s, 2H)^[21]

Compd.

Apearance

Yield/%

¹H NMR

White solid

¹H NMR (400 MHz, DMSO-d₆)δ10.13(s, 1H), 9.43(s, 1H), 8.10(d, J=8.0 Hz, 2H), 7.95(s, 1H), 7.88(d, J=8.0 Hz, 2H)^[15]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ 14.09(s, 1H), 13.83(s, 1H), 8.12(d, J=8.4 Hz, 2H), 7.91(d, J=8.4 Hz, 2H)^[16]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ 8.19(d, J=8.8 Hz, 2H), 7.88(s, 2H), 3.21(d, J=1.6 Hz, 2H), 1.34(t, J=7.2 Hz, 3H)^[17]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ10.47(s, 1H), 9.37(s, 1H), 7.91(d, J=8.4 Hz, 2H), 7.70(s, 1H), 7.57(d, J=8.4 Hz, 2H)^[18]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ 13.93(s, 1H), 13.75(s, 1H), 7.92(d, J=8.4 Hz, 2H), 7.61(d, J=8.4 Hz, 2H)^[19]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ7.98(d, J=8 Hz, 2H), 7.57 (d, J=12 Hz, 2H), 3.36-3.18(m, 2H), 1.34(t, J=7.4 Hz, 3H)^[20]

White solid

¹H NMR (400 MHz, DMSO-d₆)δ 8.00(d, J=8.4 Hz, 2H), 7.60(s, 2H), 7.42(d, J=7.2 Hz, 2H), 7.32(t, J=7.2 Hz, 2H), 7.42(d, J=7.2 Hz, 1H), 4.45(s, 2H)^[21]

Compd.	Apearance	Yield/%	mp/℃	¹H NMR, ¹³C NMR and HR-MS
b	White solid	44	76.3-76.9	¹H NMR (400 MHz, CDCl₃)δ 8.29(d, J=8.0 Hz, 4H), 7.73(d, J=8.4 Hz, 2H), 7.68(d, J=7.2 Hz, 1H), 7.56(t, J=7.6 Hz, 2H), 3.42(q, J=7.2 Hz, 2H), 1.52(t, J=7.2 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 166.04, 162.07, 161.10, 133.73, 133.1 6, 132.09, 131.66, 130.89, 128.27, 127.38, 125.60, 125.56, 27.18, 13.96. HR-MS (ESI) C₁₈H₁₄F₃N₃OS [M-H]^- Calcd.: 378.088 8, Found: 378.089 0
c	White solid	44	79.2-80.1	¹H NMR (400 MHz, CDCl₃)δ 8.78(t, J₁=7.2 Hz, J₂=1.6 Hz, 2H), 8.12(t, J₁=1.6 Hz, J₂=6.8 Hz, 2H), 7.68(t, J₁=7.6 Hz, J₂=7.6 Hz, 1H), 7.56(t, J₁=8.0 Hz, J₂=7.6 Hz, 2H), 7.45(d, J=8.8 Hz, 2H), 3.44-3.38(m, 2H), 1.52(t, J₁=7.6 Hz, J₂=7.2 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 166.03, 161.82, 161.44, 136.31, 133.62, 131.66, 131.01, 128.88, 128.42, 128.28, 128.23, 27.15, 13.99. HR-MS (ESI) C₁₇H₁₄ClN₃OS [M-H]^- Calcd.: 344.062 4, Found: 344.062 7
d	White solid	34	114.7-115.6	¹H NMR (400 MHz, CDCl₃)δ 8.31(d, J=8.0 Hz, 2H), 8.21(d, J=3.6 Hz, 1H), 7.85(s, 1H), 7.76(d, J=8.0 Hz, 2H), 6.71(d, J=2.0 Hz, 1H), 3.39(q, J=7.2 Hz, 2H), 1.54(t, J=7.2 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 161.98, 161.42, 154.54, 149.10, 144.07, 133.07, 132.18, 127.35, 125.65, 125.61, 124.97, 112.80, 27.06, 13.93. HR-MS (ESI) C₁₆H₁₃F₃N₃O₂S [M-H]^- Calcd.: 368.068 1, Found: 368.068 5
e	White solid	40	127.0-127.5	¹H NMR (400 MHz, CDCl₃)δ 8.21(d, J=3.6 Hz, 1H), 8.15(d, J=8.8 Hz, 2H), 7.85(s, 1H), 7.48(d, J=8.4 Hz, 2H), 6.70(dd, J₁=3.6 Hz, J₂=1.6 Hz, 1H)3.36(q, J=7.2 Hz, 2H), 1.52(t, J=7.2 Hz, 3H).¹³C NMR (100 MHz, CDCl₃)δ 161.76, 161.74, 154.58, 148.98, 144.16, 136.42, 128.94, 128.38, 128.20, 124.89, 112.76, 27.03, 13.97. HR-MS (ESI) C₁₅H₁₂ClN₃O₂S [M-H]^- Calcd.: 334.041 7, Found: 334.041 5
f	White solid	37	94.9-96.5	¹H NMR (400 MHz, CDCl₃)δ 8.26(d, J=8.0 Hz, 2H), 7.71(d, J=8.0 Hz, 2H), 3.33(q, J=7.6 Hz, 2H), 3.16(q, J=7.6 Hz, 2H), 1.51(t, J=7.6 Hz, 3H), 1.31(t, J=7.6 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 173.04, 160.86, 159.87, 133.22, 132.03, 131.71, 127.25, 125.59, 125.55, 28.70, 26.84, 13.98, 7.88. HR-MS (ESI) C₁₄H₁₅F₃N₃OS [M-H]^-Calcd.: 330.088 8, Found: 330.089 2
g	White solid	41	116.1-116.7	¹H NMR (400 MHz, CDCl₃)δ 8.08(d, J=8.4 Hz, 2H), 7.43(d, J=8.8 Hz, 2H), 3.32(q, J=7.6 Hz, 2H), 3.14(q, J=7.6 Hz, 2H), 1.48(t, J=7.6 Hz, 3H), 1.29(t, J=7.6 Hz, 3H). ¹³C NMR (100 MHz, CDCl₃)δ 173.04, 161.19, 159.62, 136.22, 128.87, 128.35, 138.29, 28.69, 26.80, 14.00, 7.91. HR-MS (ESI) C₁₃H₁₅ClN₃OS [M-H]^- Calcd.: 296.062 4, Found: 296.062 2
h	White solid	38	158.3-159.5	¹H NMR (400 MHz, CDCl₃)δ 8.25(d, J=7.2 Hz, 2H), 8.11(d, J=8.8 Hz, 2H), 7.65(t, J=7.6 Hz, 1H), 7.53(t, J=8.0 Hz, 4H), 7.43(d, J=8.4 Hz, 2H), 7.35(t, J=7.2 Hz, 2H), 7.29(d, J=7.2 Hz, 1H), 4.61(s, 2H). ¹³C NMR (100 MHz, CDCl₃)δ165.97, 161.40, 136.41, 136.34, 133.72, 131.69, 130.80, 130.61, 129.46, 128.97, 128.69, 128.43, 128.26, 128.24, 127.72, 37.30. HR-MS (ESI) C₂₂H₁₇ClN₃OS [M-H]^- Calcd.: 406.078 1, Found: 406.078 2
i	White solid	37	163.6-165.3	¹H NMR (400 MHz, CDCl₃)δ 8.18(d, J=3.2 Hz, 1H), 8.14(d, J=8.4 Hz, 2H), 7.82(s, 1H), 7.51(s, 1H), 7.49(q, J=3.2 Hz, 2H), 7.47(s, 1H), 7.34(t, J=7.2 Hz, 2H), 7.29(d, J=7.2 Hz, 1H), 6.68(m, 1H), 4.60(s, 2H). ¹³C NMR (100 MHz, CDCl₃)δ 161.72, 161.28, 154.52, 149.09, 144.02, 136.52, 136.27, 129.43, 129.03, 128.66, 128.39, 128.16, 127.70, 125.01, 112.79, 37.15. HR-MS (ESI) C₂₀H₁₄ClN₃O₂S [M-H]^- Calcd.: 396.057 3, Found: 396.057 0

Compd.

Apearance

Yield/%

mp/℃

¹H NMR, ¹³C NMR and HR-MS

White solid

76.3-76.9

White solid

79.2-80.1

White solid

114.7-115.6

White solid

127.0-127.5

White solid

94.9-96.5

White solid

116.1-116.7

White solid

158.3-159.5

White solid

163.6-165.3


a	Cl		CH₃	58.87	5.84±0.01
b	CF₃	CH₂CH₃	75.00	2.31±0.15
c	Cl	CH₂CH₃	74.42	3.18±0.11
d	CF₃	CH₂CH₃	81.98	1.50±0.04
e	Cl	CH₂CH₃	75.57	2.85±0.38
f	CF₃	CH₂CH₃	74.81	2.94±0.37
g	Cl	CH₂CH₃	65.76	4.68±0.12
h	Cl	Bn	30.01	> 10
i	Cl	Bn	37.89	> 10
Brequinar	-	-	-	-	0.01±0.00

Compd.

R¹

R²

R³

Inhibition rate (10 μmol·L^-1)/%

IC₅₀/μmol·L^-1a

CH₃

58.87

5.84±0.01

CF₃

CH₂CH₃

75.00

2.31±0.15

CH₂CH₃

74.42

3.18±0.11

CF₃

CH₂CH₃

81.98

1.50±0.04

CH₂CH₃

75.57

2.85±0.38

CF₃

CH₂CH₃

74.81

2.94±0.37

CH₂CH₃

65.76

4.68±0.12

30.01

> 10

37.89

> 10

Brequinar

0.01±0.00