药学学报

Compound	NMR spectrum and HRMS result	Property	Melting point/℃	Yield
SDFZ-E1	¹H NMR (400 MHz, DMSO-d₆ (dimethyl sulfoxide-d₆)) δ 11.73 (s, 1H), 8.26-8.19 (m, 1H), 8.18-8.08 (m, 2H), 8.02 (t, J = 6.6 Hz, 2H), 7.95 (s, 1H), 7.91-7.80 (m, 2H), 7.61 (p, J = 7.4 Hz, 4H), 6.01 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 133.8, 132.1, 131.6, 131.6, 129.7, 129.0, 128.2, 127.6, 127.1, 126.6, 125.9, 124.3, 122.3, 121.7, 100.4, 70.0. HRMS (ESI⁺): m/z calculated for C₂₁H₁₆N₂O₃ 345.123 4, found [M+H]⁺ 345.121 0.	White solid	250-252	15%
SDFZ-E2	¹H NMR (400 MHz, DMSO-d₆) δ 11.77 (s, 1H), 8.25 (d, J = 8.4 Hz, 1H), 8.13 (d, J = 8.5 Hz, 1H), 7.90 (t, J = 7.7 Hz, 1H), 7.76 (s, 1H), 7.71 (t, J = 7.7 Hz, 1H), 7.60 (d, J = 7.5 Hz, 2H), 7.52-7.36 (m, 3H), 5.55 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 136.3, 136.3, 131.4, 131.4, 129.8, 129.1, 128.7, 128.7, 128.1, 122.2, 70.8. HRMS (ESI⁺): m/z calculated for C₁₇H₁₄N₂O₃ 295.107 7, found [M+H]⁺ 295.105 2.	White solid	248-250	18%
SDFZ-E3	¹H NMR (400 MHz, DMSO-d₆) δ 11.48 (s, 1H), 9.20 (s, 1H), 8.23 (d, J = 8.3 Hz, 1H), 8.04 (d, J = 8.4 Hz, 1H), 7.84 (q, J = 6.9, 6.3 Hz, 1H), 7.66 (q, J = 6.6 Hz, 4H), 7.56 (d, J = 8.0 Hz, 2H), 5.49 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 163.2, 162.0, 151.9, 147.7, 139.2, 131.5, 131.3, 131.3, 131.1, 130.7, 129.5, 127.7, 127.6, 127.1, 127.1, 122.2, 122.1, 121.6, 99.5, 69.6. HRMS (ESI⁺): m/z calculated for C₁₇H₁₃BrN₂O₃ 373.018 2, found [M+H]⁺ 373.018 0.	Yellowish solid	220-222	22%
SDFZ-E4	¹H NMR (400 MHz, DMSO-d₆) δ 11.90 (s, 1H), 8.18 (dd, J = 23.3, 8.7 Hz, 2H), 8.04-7.63 (m, 4H), 7.59-7.51 (m, 2H), 7.07-6.99 (m, 2H), 5.49 (d, J = 5.3 Hz, 2H), 3.79 (s, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 159.9, 159.7, 140.6, 133.4, 130.8, 130.4, 129.6, 128.3, 124.6, 114.5, 114.4, 106.0, 100.5, 71.7, 71.2, 55.6, 55.5, 46.7. HRMS (ESI⁺): m/z calculated for C₁₈H₁₆N₂O₄ 325.118 3, found [M+H]⁺ 325.118 8.	White solid	248-250	16%
SDFZ-E5	¹H NMR (400 MHz, DMSO-d₆) δ 11.85 (s, 1H), 8.24 (d, J = 8.4 Hz, 1H), 8.16 (d, J = 8.5 Hz, 1H), 7.92 (t, J = 7.8 Hz, 1H), 7.82 (d, J = 3.4 Hz, 1H), 7.71 (t, J = 7.7 Hz, 1H), 7.57-7.45 (m, 4H), 5.52 (s, 2H), 1.31 (d, J = 1.9 Hz, 9H). ¹³C NMR (101 MHz, DMSO-d₆) δ 151.4, 132.8, 128.4, 125.9, 122.6, 121.5, 100.4, 71.7, 34.8, 31.5. HRMS (ESI⁺): m/z calculated for C₂₁H₂₂N₂O₃ 351.170 3, found [M+H]⁺ 351.167 7.	White solid	256-258	32%
SDFZ-E6	¹H NMR (400 MHz, DMSO-d₆) δ 11.58 (s, 1H), 8.24 (d, J = 8.4 Hz, 1H), 8.10 (d, J = 9.5 Hz, 1H), 7.88 (t, J = 7.4 Hz, 1H), 7.82-7.40 (m, 6H), 5.54 (t, J = 4.4 Hz, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 163.2, 162.1, 151.9, 147.7, 136.0, 132.0, 131.1, 130.2, 129.5, 127.6, 122.1, 121.8, 121.7, 99.6, 69.7. HRMS (ESI⁺): m/z calculated for C₁₇H₁₃BrN₂O₃ 373.018 2, found [M+H]⁺ 373.017 2.	White solid	286-288	16%
SDFZ-E7	¹H NMR (400 MHz, DMSO-d₆) δ 11.67 (s, 1H), 8.25 (d, J = 8.4 Hz, 1H), 8.11 (dd, J = 8.5, 3.4 Hz, 1H), 7.89 (t, J = 7.9 Hz, 1H), 7.84-7.63 (m, 4H), 7.57 (d, J = 8.1 Hz, 2H), 5.53 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 135.6, 132.1, 132.0, 131.8, 130.4, 128.2, 122.4, 122.0, 121.6, 100.2, 70.4, 57.8. HRMS (ESI⁺): m/z calculated for C₁₇H₁₃ClN₂O₃ 329.068 8, found [M+H]⁺ 329.067 4.	White solid	214-216	18%
SDFZ-E8	¹H NMR (400 MHz, DMSO-d₆) δ 11.79 (s, 1H), 8.26 (d, J = 9.2 Hz, 2H), 7.96 (s, 1H), 7.76 (s, 2H), 4.70-4.42 (m, 2H), 1.53 (dt, J = 9.2, 4.7 Hz, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 166.1, 158.8, 149.0, 143.0, 133.5, 128.7, 125.2, 122.7, 121.3, 100.4, 67.1, 14.6. HRMS (ESI⁺): m/z calculated for C₁₂H₁₂N₂O₃ 233.092 1, found [M+H]⁺ 233.091 0.	White solid	194-196	23%
SDFZ-E9	¹H NMR (400 MHz, DMSO-d₆) δ 11.65 (s, 1H), 8.25-8.02 (m, 3H), 7.87 (d, J = 8.4 Hz, 2H), 7.77 (s, 1H), 7.64 (t, J = 7.6 Hz, 1H), 7.46 (dq, J = 24.6, 7.8 Hz, 4H), 7.13 (d, J = 7.5 Hz, 1H), 4.91 (d, J = 4.9 Hz, 2H), 4.71 (d, J = 3.6 Hz, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 154.2, 134.5, 127.9, 126.9, 126.6, 125.8, 125.4, 122.3, 121.9, 121.6, 120.8, 106.3, 99.9, 68.7, 67.1. HRMS (ESI⁺): m/z calculated for C₂₂H₁₈N₂O₄ 375.133 9, found [M+H]⁺ 375.131 8.	Yellowish solid	192-194	14%
SDFZ-E10	¹H NMR (400 MHz, DMSO-d₆) δ 11.69 (s, 1H), 8.21 (d, J = 8.5 Hz, 1H), 8.10 (d, J = 8.5 Hz, 1H), 7.93-7.85 (m, 1H), 7.70 (t, J = 7.7 Hz, 1H), 7.61 (s, 1H), 4.17 (s, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 145.4, 144.9, 134.2, 133.6, 132.8, 132.2, 131.9, 5.3. HRMS (ESI⁺): m/z calculated for C₁₁H₁₀N₂O₃ 219.076 4, found [M+H]⁺ 219.075 2.	White solid	238-240	20%
SDFZ-E11	¹H NMR (400 MHz, DMSO-d₆) δ 10.92 (s, 1H), 8.34 – 8.20 (m, 3H), 7.93 (dd, J = 8.8, 6.7 Hz, 2H), 7.74 (h, J = 8.2 Hz, 5H), 7.55-7.32 (m, 4H), 4.19 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 131.9, 131.4, 128.9, 128.4, 128.2, 128.1, 127.5, 127.2, 125.2, 124.4, 122.2, 121.8, 99.0, 57.3, 57.1. HRMS (ESI⁺): m/z calculated for C₂₃H₁₈N₂O₃ 371.139 0, found [M+H]⁺ 371.139 4.	White solid	> 300	13%
SDFZ-E12	¹H NMR (400 MHz, DMSO-d₆) δ 11.95 (s, 1H), 8.19 (dt, J = 8.4, 2.2 Hz, 2H), 7.99-7.90 (m, 1H), 7.83-7.71 (m, 2H), 7.44 (d, J = 7.1 Hz, 2H), 7.34 (t, J = 7.6 Hz, 2H), 7.28-7.22 (m, 1H), 4.68 (t, J = 6.4 Hz, 2H), 3.26 (t, J = 6.4 Hz, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 165.6, 159.1, 149.4, 138.3, 133.3, 129.5, 128.8, 128.7, 126.9, 125.7, 122.5, 121.3, 100.4, 71.3, 34.8. HRMS (ESI⁺): m/z calculated for C₁₈H₁₆N₂O₃ 309.123 4, found [M+H]⁺ 309.121 9.	White solid	180-182	21%
SDFZ-E13	¹H NMR (400 MHz, DMSO-d₆) δ 11.73 (s, 1H), 8.21-8.04 (m, 2H), 7.89 (t, J = 7.4 Hz, 1H), 7.69 (d, J = 6.9 Hz, 2H), 6.91 (s, 2H), 4.86-4.67 (m, 2H), 4.62-4.46 (m, 2H), 2.30 (s, 6H). ¹³C NMR (101 MHz, DMSO-d₆) δ 156.8, 137.1, 128.2, 125.8, 122.4, 121.4, 115.4, 100.0, 69.1, 66.5, 20.8. HRMS (ESI⁺): m/z calculated for C₂₀H₁₉ClN₂O₄ 387.110 6, found [M+H]⁺ 387.109 5.	White solid	216-218	17%

Compound

NMR spectrum and HRMS result

Property

Melting point/℃

Yield

SDFZ-E1

¹H NMR (400 MHz, DMSO-d₆ (dimethyl sulfoxide-d₆)) δ 11.73 (s, 1H), 8.26-8.19 (m, 1H), 8.18-8.08 (m, 2H), 8.02 (t, J = 6.6 Hz, 2H), 7.95 (s, 1H), 7.91-7.80 (m, 2H), 7.61 (p, J = 7.4 Hz, 4H), 6.01 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 133.8, 132.1, 131.6, 131.6, 129.7, 129.0, 128.2, 127.6, 127.1, 126.6, 125.9, 124.3, 122.3, 121.7, 100.4, 70.0. HRMS (ESI⁺): m/z calculated for C₂₁H₁₆N₂O₃ 345.123 4, found [M+H]⁺ 345.121 0.

White solid

250-252

15%

SDFZ-E2

¹H NMR (400 MHz, DMSO-d₆) δ 11.77 (s, 1H), 8.25 (d, J = 8.4 Hz, 1H), 8.13 (d, J = 8.5 Hz, 1H), 7.90 (t, J = 7.7 Hz, 1H), 7.76 (s, 1H), 7.71 (t, J = 7.7 Hz, 1H), 7.60 (d, J = 7.5 Hz, 2H), 7.52-7.36 (m, 3H), 5.55 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 136.3, 136.3, 131.4, 131.4, 129.8, 129.1, 128.7, 128.7, 128.1, 122.2, 70.8. HRMS (ESI⁺): m/z calculated for C₁₇H₁₄N₂O₃ 295.107 7, found [M+H]⁺ 295.105 2.

White solid

248-250

18%

SDFZ-E3

¹H NMR (400 MHz, DMSO-d₆) δ 11.48 (s, 1H), 9.20 (s, 1H), 8.23 (d, J = 8.3 Hz, 1H), 8.04 (d, J = 8.4 Hz, 1H), 7.84 (q, J = 6.9, 6.3 Hz, 1H), 7.66 (q, J = 6.6 Hz, 4H), 7.56 (d, J = 8.0 Hz, 2H), 5.49 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 163.2, 162.0, 151.9, 147.7, 139.2, 131.5, 131.3, 131.3, 131.1, 130.7, 129.5, 127.7, 127.6, 127.1, 127.1, 122.2, 122.1, 121.6, 99.5, 69.6. HRMS (ESI⁺): m/z calculated for C₁₇H₁₃BrN₂O₃ 373.018 2, found [M+H]⁺ 373.018 0.

Yellowish solid

220-222

22%

SDFZ-E4

¹H NMR (400 MHz, DMSO-d₆) δ 11.90 (s, 1H), 8.18 (dd, J = 23.3, 8.7 Hz, 2H), 8.04-7.63 (m, 4H), 7.59-7.51 (m, 2H), 7.07-6.99 (m, 2H), 5.49 (d, J = 5.3 Hz, 2H), 3.79 (s, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 159.9, 159.7, 140.6, 133.4, 130.8, 130.4, 129.6, 128.3, 124.6, 114.5, 114.4, 106.0, 100.5, 71.7, 71.2, 55.6, 55.5, 46.7. HRMS (ESI⁺): m/z calculated for C₁₈H₁₆N₂O₄ 325.118 3, found [M+H]⁺ 325.118 8.

White solid

248-250

16%

SDFZ-E5

¹H NMR (400 MHz, DMSO-d₆) δ 11.85 (s, 1H), 8.24 (d, J = 8.4 Hz, 1H), 8.16 (d, J = 8.5 Hz, 1H), 7.92 (t, J = 7.8 Hz, 1H), 7.82 (d, J = 3.4 Hz, 1H), 7.71 (t, J = 7.7 Hz, 1H), 7.57-7.45 (m, 4H), 5.52 (s, 2H), 1.31 (d, J = 1.9 Hz, 9H). ¹³C NMR (101 MHz, DMSO-d₆) δ 151.4, 132.8, 128.4, 125.9, 122.6, 121.5, 100.4, 71.7, 34.8, 31.5. HRMS (ESI⁺): m/z calculated for C₂₁H₂₂N₂O₃ 351.170 3, found [M+H]⁺ 351.167 7.

White solid

256-258

32%

SDFZ-E6

¹H NMR (400 MHz, DMSO-d₆) δ 11.58 (s, 1H), 8.24 (d, J = 8.4 Hz, 1H), 8.10 (d, J = 9.5 Hz, 1H), 7.88 (t, J = 7.4 Hz, 1H), 7.82-7.40 (m, 6H), 5.54 (t, J = 4.4 Hz, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 163.2, 162.1, 151.9, 147.7, 136.0, 132.0, 131.1, 130.2, 129.5, 127.6, 122.1, 121.8, 121.7, 99.6, 69.7. HRMS (ESI⁺): m/z calculated for C₁₇H₁₃BrN₂O₃ 373.018 2, found [M+H]⁺ 373.017 2.

White solid

286-288

16%

SDFZ-E7

¹H NMR (400 MHz, DMSO-d₆) δ 11.67 (s, 1H), 8.25 (d, J = 8.4 Hz, 1H), 8.11 (dd, J = 8.5, 3.4 Hz, 1H), 7.89 (t, J = 7.9 Hz, 1H), 7.84-7.63 (m, 4H), 7.57 (d, J = 8.1 Hz, 2H), 5.53 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 135.6, 132.1, 132.0, 131.8, 130.4, 128.2, 122.4, 122.0, 121.6, 100.2, 70.4, 57.8. HRMS (ESI⁺): m/z calculated for C₁₇H₁₃ClN₂O₃ 329.068 8, found [M+H]⁺ 329.067 4.

White solid

214-216

18%

SDFZ-E8

¹H NMR (400 MHz, DMSO-d₆) δ 11.79 (s, 1H), 8.26 (d, J = 9.2 Hz, 2H), 7.96 (s, 1H), 7.76 (s, 2H), 4.70-4.42 (m, 2H), 1.53 (dt, J = 9.2, 4.7 Hz, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 166.1, 158.8, 149.0, 143.0, 133.5, 128.7, 125.2, 122.7, 121.3, 100.4, 67.1, 14.6. HRMS (ESI⁺): m/z calculated for C₁₂H₁₂N₂O₃ 233.092 1, found [M+H]⁺ 233.091 0.

White solid

194-196

23%

SDFZ-E9

¹H NMR (400 MHz, DMSO-d₆) δ 11.65 (s, 1H), 8.25-8.02 (m, 3H), 7.87 (d, J = 8.4 Hz, 2H), 7.77 (s, 1H), 7.64 (t, J = 7.6 Hz, 1H), 7.46 (dq, J = 24.6, 7.8 Hz, 4H), 7.13 (d, J = 7.5 Hz, 1H), 4.91 (d, J = 4.9 Hz, 2H), 4.71 (d, J = 3.6 Hz, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 154.2, 134.5, 127.9, 126.9, 126.6, 125.8, 125.4, 122.3, 121.9, 121.6, 120.8, 106.3, 99.9, 68.7, 67.1. HRMS (ESI⁺): m/z calculated for C₂₂H₁₈N₂O₄ 375.133 9, found [M+H]⁺ 375.131 8.

Yellowish solid

192-194

14%

SDFZ-E10

¹H NMR (400 MHz, DMSO-d₆) δ 11.69 (s, 1H), 8.21 (d, J = 8.5 Hz, 1H), 8.10 (d, J = 8.5 Hz, 1H), 7.93-7.85 (m, 1H), 7.70 (t, J = 7.7 Hz, 1H), 7.61 (s, 1H), 4.17 (s, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 145.4, 144.9, 134.2, 133.6, 132.8, 132.2, 131.9, 5.3. HRMS (ESI⁺): m/z calculated for C₁₁H₁₀N₂O₃ 219.076 4, found [M+H]⁺ 219.075 2.

White solid

238-240

20%

SDFZ-E11

¹H NMR (400 MHz, DMSO-d₆) δ 10.92 (s, 1H), 8.34 – 8.20 (m, 3H), 7.93 (dd, J = 8.8, 6.7 Hz, 2H), 7.74 (h, J = 8.2 Hz, 5H), 7.55-7.32 (m, 4H), 4.19 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 131.9, 131.4, 128.9, 128.4, 128.2, 128.1, 127.5, 127.2, 125.2, 124.4, 122.2, 121.8, 99.0, 57.3, 57.1. HRMS (ESI⁺): m/z calculated for C₂₃H₁₈N₂O₃ 371.139 0, found [M+H]⁺ 371.139 4.

White solid

> 300

13%

SDFZ-E12

¹H NMR (400 MHz, DMSO-d₆) δ 11.95 (s, 1H), 8.19 (dt, J = 8.4, 2.2 Hz, 2H), 7.99-7.90 (m, 1H), 7.83-7.71 (m, 2H), 7.44 (d, J = 7.1 Hz, 2H), 7.34 (t, J = 7.6 Hz, 2H), 7.28-7.22 (m, 1H), 4.68 (t, J = 6.4 Hz, 2H), 3.26 (t, J = 6.4 Hz, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 165.6, 159.1, 149.4, 138.3, 133.3, 129.5, 128.8, 128.7, 126.9, 125.7, 122.5, 121.3, 100.4, 71.3, 34.8. HRMS (ESI⁺): m/z calculated for C₁₈H₁₆N₂O₃ 309.123 4, found [M+H]⁺ 309.121 9.

White solid

180-182

21%

SDFZ-E13

¹H NMR (400 MHz, DMSO-d₆) δ 11.73 (s, 1H), 8.21-8.04 (m, 2H), 7.89 (t, J = 7.4 Hz, 1H), 7.69 (d, J = 6.9 Hz, 2H), 6.91 (s, 2H), 4.86-4.67 (m, 2H), 4.62-4.46 (m, 2H), 2.30 (s, 6H). ¹³C NMR (101 MHz, DMSO-d₆) δ 156.8, 137.1, 128.2, 125.8, 122.4, 121.4, 115.4, 100.0, 69.1, 66.5, 20.8. HRMS (ESI⁺): m/z calculated for C₂₀H₁₉ClN₂O₄ 387.110 6, found [M+H]⁺ 387.109 5.

White solid

216-218

17%

Compound	NMR spectrum and HRMS result	Property	Melting point/℃	Yield
SDFZ-E1	¹H NMR (400 MHz, DMSO-d₆ (dimethyl sulfoxide-d₆)) δ 11.73 (s, 1H), 8.26-8.19 (m, 1H), 8.18-8.08 (m, 2H), 8.02 (t, J = 6.6 Hz, 2H), 7.95 (s, 1H), 7.91-7.80 (m, 2H), 7.61 (p, J = 7.4 Hz, 4H), 6.01 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 133.8, 132.1, 131.6, 131.6, 129.7, 129.0, 128.2, 127.6, 127.1, 126.6, 125.9, 124.3, 122.3, 121.7, 100.4, 70.0. HRMS (ESI⁺): m/z calculated for C₂₁H₁₆N₂O₃ 345.123 4, found [M+H]⁺ 345.121 0.	White solid	250-252	15%
SDFZ-E2	¹H NMR (400 MHz, DMSO-d₆) δ 11.77 (s, 1H), 8.25 (d, J = 8.4 Hz, 1H), 8.13 (d, J = 8.5 Hz, 1H), 7.90 (t, J = 7.7 Hz, 1H), 7.76 (s, 1H), 7.71 (t, J = 7.7 Hz, 1H), 7.60 (d, J = 7.5 Hz, 2H), 7.52-7.36 (m, 3H), 5.55 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 136.3, 136.3, 131.4, 131.4, 129.8, 129.1, 128.7, 128.7, 128.1, 122.2, 70.8. HRMS (ESI⁺): m/z calculated for C₁₇H₁₄N₂O₃ 295.107 7, found [M+H]⁺ 295.105 2.	White solid	248-250	18%
SDFZ-E3	¹H NMR (400 MHz, DMSO-d₆) δ 11.48 (s, 1H), 9.20 (s, 1H), 8.23 (d, J = 8.3 Hz, 1H), 8.04 (d, J = 8.4 Hz, 1H), 7.84 (q, J = 6.9, 6.3 Hz, 1H), 7.66 (q, J = 6.6 Hz, 4H), 7.56 (d, J = 8.0 Hz, 2H), 5.49 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 163.2, 162.0, 151.9, 147.7, 139.2, 131.5, 131.3, 131.3, 131.1, 130.7, 129.5, 127.7, 127.6, 127.1, 127.1, 122.2, 122.1, 121.6, 99.5, 69.6. HRMS (ESI⁺): m/z calculated for C₁₇H₁₃BrN₂O₃ 373.018 2, found [M+H]⁺ 373.018 0.	Yellowish solid	220-222	22%
SDFZ-E4	¹H NMR (400 MHz, DMSO-d₆) δ 11.90 (s, 1H), 8.18 (dd, J = 23.3, 8.7 Hz, 2H), 8.04-7.63 (m, 4H), 7.59-7.51 (m, 2H), 7.07-6.99 (m, 2H), 5.49 (d, J = 5.3 Hz, 2H), 3.79 (s, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 159.9, 159.7, 140.6, 133.4, 130.8, 130.4, 129.6, 128.3, 124.6, 114.5, 114.4, 106.0, 100.5, 71.7, 71.2, 55.6, 55.5, 46.7. HRMS (ESI⁺): m/z calculated for C₁₈H₁₆N₂O₄ 325.118 3, found [M+H]⁺ 325.118 8.	White solid	248-250	16%
SDFZ-E5	¹H NMR (400 MHz, DMSO-d₆) δ 11.85 (s, 1H), 8.24 (d, J = 8.4 Hz, 1H), 8.16 (d, J = 8.5 Hz, 1H), 7.92 (t, J = 7.8 Hz, 1H), 7.82 (d, J = 3.4 Hz, 1H), 7.71 (t, J = 7.7 Hz, 1H), 7.57-7.45 (m, 4H), 5.52 (s, 2H), 1.31 (d, J = 1.9 Hz, 9H). ¹³C NMR (101 MHz, DMSO-d₆) δ 151.4, 132.8, 128.4, 125.9, 122.6, 121.5, 100.4, 71.7, 34.8, 31.5. HRMS (ESI⁺): m/z calculated for C₂₁H₂₂N₂O₃ 351.170 3, found [M+H]⁺ 351.167 7.	White solid	256-258	32%
SDFZ-E6	¹H NMR (400 MHz, DMSO-d₆) δ 11.58 (s, 1H), 8.24 (d, J = 8.4 Hz, 1H), 8.10 (d, J = 9.5 Hz, 1H), 7.88 (t, J = 7.4 Hz, 1H), 7.82-7.40 (m, 6H), 5.54 (t, J = 4.4 Hz, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 163.2, 162.1, 151.9, 147.7, 136.0, 132.0, 131.1, 130.2, 129.5, 127.6, 122.1, 121.8, 121.7, 99.6, 69.7. HRMS (ESI⁺): m/z calculated for C₁₇H₁₃BrN₂O₃ 373.018 2, found [M+H]⁺ 373.017 2.	White solid	286-288	16%
SDFZ-E7	¹H NMR (400 MHz, DMSO-d₆) δ 11.67 (s, 1H), 8.25 (d, J = 8.4 Hz, 1H), 8.11 (dd, J = 8.5, 3.4 Hz, 1H), 7.89 (t, J = 7.9 Hz, 1H), 7.84-7.63 (m, 4H), 7.57 (d, J = 8.1 Hz, 2H), 5.53 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 135.6, 132.1, 132.0, 131.8, 130.4, 128.2, 122.4, 122.0, 121.6, 100.2, 70.4, 57.8. HRMS (ESI⁺): m/z calculated for C₁₇H₁₃ClN₂O₃ 329.068 8, found [M+H]⁺ 329.067 4.	White solid	214-216	18%
SDFZ-E8	¹H NMR (400 MHz, DMSO-d₆) δ 11.79 (s, 1H), 8.26 (d, J = 9.2 Hz, 2H), 7.96 (s, 1H), 7.76 (s, 2H), 4.70-4.42 (m, 2H), 1.53 (dt, J = 9.2, 4.7 Hz, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 166.1, 158.8, 149.0, 143.0, 133.5, 128.7, 125.2, 122.7, 121.3, 100.4, 67.1, 14.6. HRMS (ESI⁺): m/z calculated for C₁₂H₁₂N₂O₃ 233.092 1, found [M+H]⁺ 233.091 0.	White solid	194-196	23%
SDFZ-E9	¹H NMR (400 MHz, DMSO-d₆) δ 11.65 (s, 1H), 8.25-8.02 (m, 3H), 7.87 (d, J = 8.4 Hz, 2H), 7.77 (s, 1H), 7.64 (t, J = 7.6 Hz, 1H), 7.46 (dq, J = 24.6, 7.8 Hz, 4H), 7.13 (d, J = 7.5 Hz, 1H), 4.91 (d, J = 4.9 Hz, 2H), 4.71 (d, J = 3.6 Hz, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 154.2, 134.5, 127.9, 126.9, 126.6, 125.8, 125.4, 122.3, 121.9, 121.6, 120.8, 106.3, 99.9, 68.7, 67.1. HRMS (ESI⁺): m/z calculated for C₂₂H₁₈N₂O₄ 375.133 9, found [M+H]⁺ 375.131 8.	Yellowish solid	192-194	14%
SDFZ-E10	¹H NMR (400 MHz, DMSO-d₆) δ 11.69 (s, 1H), 8.21 (d, J = 8.5 Hz, 1H), 8.10 (d, J = 8.5 Hz, 1H), 7.93-7.85 (m, 1H), 7.70 (t, J = 7.7 Hz, 1H), 7.61 (s, 1H), 4.17 (s, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 145.4, 144.9, 134.2, 133.6, 132.8, 132.2, 131.9, 5.3. HRMS (ESI⁺): m/z calculated for C₁₁H₁₀N₂O₃ 219.076 4, found [M+H]⁺ 219.075 2.	White solid	238-240	20%
SDFZ-E11	¹H NMR (400 MHz, DMSO-d₆) δ 10.92 (s, 1H), 8.34 – 8.20 (m, 3H), 7.93 (dd, J = 8.8, 6.7 Hz, 2H), 7.74 (h, J = 8.2 Hz, 5H), 7.55-7.32 (m, 4H), 4.19 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 131.9, 131.4, 128.9, 128.4, 128.2, 128.1, 127.5, 127.2, 125.2, 124.4, 122.2, 121.8, 99.0, 57.3, 57.1. HRMS (ESI⁺): m/z calculated for C₂₃H₁₈N₂O₃ 371.139 0, found [M+H]⁺ 371.139 4.	White solid	> 300	13%
SDFZ-E12	¹H NMR (400 MHz, DMSO-d₆) δ 11.95 (s, 1H), 8.19 (dt, J = 8.4, 2.2 Hz, 2H), 7.99-7.90 (m, 1H), 7.83-7.71 (m, 2H), 7.44 (d, J = 7.1 Hz, 2H), 7.34 (t, J = 7.6 Hz, 2H), 7.28-7.22 (m, 1H), 4.68 (t, J = 6.4 Hz, 2H), 3.26 (t, J = 6.4 Hz, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 165.6, 159.1, 149.4, 138.3, 133.3, 129.5, 128.8, 128.7, 126.9, 125.7, 122.5, 121.3, 100.4, 71.3, 34.8. HRMS (ESI⁺): m/z calculated for C₁₈H₁₆N₂O₃ 309.123 4, found [M+H]⁺ 309.121 9.	White solid	180-182	21%
SDFZ-E13	¹H NMR (400 MHz, DMSO-d₆) δ 11.73 (s, 1H), 8.21-8.04 (m, 2H), 7.89 (t, J = 7.4 Hz, 1H), 7.69 (d, J = 6.9 Hz, 2H), 6.91 (s, 2H), 4.86-4.67 (m, 2H), 4.62-4.46 (m, 2H), 2.30 (s, 6H). ¹³C NMR (101 MHz, DMSO-d₆) δ 156.8, 137.1, 128.2, 125.8, 122.4, 121.4, 115.4, 100.0, 69.1, 66.5, 20.8. HRMS (ESI⁺): m/z calculated for C₂₀H₁₉ClN₂O₄ 387.110 6, found [M+H]⁺ 387.109 5.	White solid	216-218	17%

Compound

NMR spectrum and HRMS result

Property

Melting point/℃

Yield

SDFZ-E1

White solid

250-252

15%

SDFZ-E2

White solid

248-250

18%

SDFZ-E3

Yellowish solid

220-222

22%

SDFZ-E4

White solid

248-250

16%

SDFZ-E5

White solid

256-258

32%

SDFZ-E6

White solid

286-288

16%

SDFZ-E7

White solid

214-216

18%

SDFZ-E8

White solid

194-196

23%

SDFZ-E9

Yellowish solid

192-194

14%

SDFZ-E10

White solid

238-240

20%

SDFZ-E11

White solid

> 300

13%

SDFZ-E12

White solid

180-182

21%

SDFZ-E13

White solid

216-218

17%


SDFZ-E1	-1-Napthene	> 50
SDFZ-E2	-Ph	2.49 ± 0.49
SDFZ-E3	-3-Br-Ph	7.45 ± 0.26
SDFZ-E4	-4-Methoxy-Ph	32.98 ± 0.29
SDFZ-E5	-4-t-Bu-Ph	> 50
SDFZ-E6	-4-Br-Ph	> 50
SDFZ-E7	-4-Cl-Ph	> 50
SDFZ-E8	-CH₃	> 50
SDFZ-E9		> 50
SDFZ-E10	H	> 50
SDFZ-E11	-4-Ph-Ph	> 50
SDFZ-E12	-Bn	12.94 ± 1.14
SDFZ-E13		> 50
SAHA	-	6.39 ± 0.62
PCI-34051	-	0.051 ± 0.016

Compound

^aIC₅₀ of HDAC8/μmol·L^-1

SDFZ-E1

-1-Napthene

> 50

SDFZ-E2

-Ph

2.49 ± 0.49

SDFZ-E3

-3-Br-Ph

7.45 ± 0.26

SDFZ-E4

-4-Methoxy-Ph

32.98 ± 0.29

SDFZ-E5

-4-t-Bu-Ph

> 50

SDFZ-E6

-4-Br-Ph

> 50

SDFZ-E7

-4-Cl-Ph

> 50

SDFZ-E8

-CH₃

> 50

SDFZ-E9

> 50

SDFZ-E10

> 50

SDFZ-E11

-4-Ph-Ph

> 50

SDFZ-E12

-Bn

12.94 ± 1.14

SDFZ-E13

> 50

SAHA

6.39 ± 0.62

PCI-34051

0.051 ± 0.016


SDFZ-E1	-1-Napthene	> 50
SDFZ-E2	-Ph	2.49 ± 0.49
SDFZ-E3	-3-Br-Ph	7.45 ± 0.26
SDFZ-E4	-4-Methoxy-Ph	32.98 ± 0.29
SDFZ-E5	-4-t-Bu-Ph	> 50
SDFZ-E6	-4-Br-Ph	> 50
SDFZ-E7	-4-Cl-Ph	> 50
SDFZ-E8	-CH₃	> 50
SDFZ-E9		> 50
SDFZ-E10	H	> 50
SDFZ-E11	-4-Ph-Ph	> 50
SDFZ-E12	-Bn	12.94 ± 1.14
SDFZ-E13		> 50
SAHA	-	6.39 ± 0.62
PCI-34051	-	0.051 ± 0.016

Compound

^aIC₅₀ of HDAC8/μmol·L^-1

SDFZ-E1

-1-Napthene

> 50

SDFZ-E2

-Ph

2.49 ± 0.49

SDFZ-E3

-3-Br-Ph

7.45 ± 0.26

SDFZ-E4

-4-Methoxy-Ph

32.98 ± 0.29

SDFZ-E5

-4-t-Bu-Ph

> 50

SDFZ-E6

-4-Br-Ph

> 50

SDFZ-E7

-4-Cl-Ph

> 50

SDFZ-E8

-CH₃

> 50

SDFZ-E9

> 50

SDFZ-E10

> 50

SDFZ-E11

-4-Ph-Ph

> 50

SDFZ-E12

-Bn

12.94 ± 1.14

SDFZ-E13

> 50

SAHA

6.39 ± 0.62

PCI-34051

0.051 ± 0.016

Compound	^aIC₅₀/μmol·L^-1
SDFZ-E2	> 50	> 50	33.12 ± 10.51	2.49 ± 0.49
SDFZ-E3	> 50	> 50	> 50	7.45 ± 0.26
SDFZ-E12	> 50	> 50	41.23 ± 3.21	12.95 ± 1.14
SAHA	0.037 ± 0.007	0.015 ± 0.002	0.090 ± 0.007	6.39 ± 0.62

Compound

^aIC₅₀/μmol·L^-1

HDAC1

HDAC2

HDAC6

HDAC8

SDFZ-E2

> 50

33.12 ± 10.51

2.49 ± 0.49

SDFZ-E3

> 50

7.45 ± 0.26

SDFZ-E12

> 50

41.23 ± 3.21

12.95 ± 1.14

SAHA

0.037 ± 0.007

0.015 ± 0.002

0.090 ± 0.007

6.39 ± 0.62

Compound	^aIC₅₀/μmol·L^-1
SDFZ-E2	> 50	> 50	33.12 ± 10.51	2.49 ± 0.49
SDFZ-E3	> 50	> 50	> 50	7.45 ± 0.26
SDFZ-E12	> 50	> 50	41.23 ± 3.21	12.95 ± 1.14
SAHA	0.037 ± 0.007	0.015 ± 0.002	0.090 ± 0.007	6.39 ± 0.62

Compound

^aIC₅₀/μmol·L^-1

HDAC1

HDAC2

HDAC6

HDAC8

SDFZ-E2

> 50

33.12 ± 10.51

2.49 ± 0.49

SDFZ-E3

> 50

7.45 ± 0.26

SDFZ-E12

> 50

41.23 ± 3.21

12.95 ± 1.14

SAHA

0.037 ± 0.007

0.015 ± 0.002

0.090 ± 0.007

6.39 ± 0.62

Compound	^aIC₅₀/μmol·L^-1
SDFZ-E2	13.99 ± 1.75	11.25 ± 0.94	42.07 ± 5.17
SDFZ-E3	9.68 ± 0.24	3.72 ± 1.12	10.65 ± 0.25
SDFZ-E12	37.44 ± 4.02	26.44 ± 5.90	> 200
PCI-34051	15.91 ± 0.58	21.12 ± 2.25	67.10 ± 9.97

Compound

^aIC₅₀/μmol·L^-1

SK-N-BE(2)

SU-DHL-2

Jurkat

SDFZ-E2

13.99 ± 1.75

11.25 ± 0.94

42.07 ± 5.17

SDFZ-E3

9.68 ± 0.24

3.72 ± 1.12

10.65 ± 0.25

SDFZ-E12

37.44 ± 4.02

26.44 ± 5.90

> 200

PCI-34051

15.91 ± 0.58

21.12 ± 2.25

67.10 ± 9.97

Compound	^aIC₅₀/μmol·L^-1
SDFZ-E2	13.99 ± 1.75	11.25 ± 0.94	42.07 ± 5.17
SDFZ-E3	9.68 ± 0.24	3.72 ± 1.12	10.65 ± 0.25
SDFZ-E12	37.44 ± 4.02	26.44 ± 5.90	> 200
PCI-34051	15.91 ± 0.58	21.12 ± 2.25	67.10 ± 9.97

Compound

^aIC₅₀/μmol·L^-1

SK-N-BE(2)

SU-DHL-2

Jurkat

SDFZ-E2

13.99 ± 1.75

11.25 ± 0.94

42.07 ± 5.17

SDFZ-E3

9.68 ± 0.24

3.72 ± 1.12

10.65 ± 0.25

SDFZ-E12

37.44 ± 4.02

26.44 ± 5.90

> 200

PCI-34051

15.91 ± 0.58

21.12 ± 2.25

67.10 ± 9.97

喹啉类HDAC8选择性抑制剂的设计、合成及抗肿瘤活性研究

PDF下载

周易 ¹ , 邵文晴 ¹ , 杨新颖 ² , 侯旭奔 ²^,^* , 方浩 ²^,^*

药学学报 | 研究论文 2024,59(4): 979-986

收起

药学学报 | 研究论文 2024, 59(4): 979-986

喹啉类HDAC8选择性抑制剂的设计、合成及抗肿瘤活性研究

全屏

周易¹, 邵文晴¹, 杨新颖², 侯旭奔²^,^*, 方浩²^,^*

作者信息

1.山东第一医科大学附属省立医院药学部, 山东济南 250021

2.山东大学药学院, 天然产物化学生物学教育部重点实验室, 山东济南 250012

通讯作者:

^*侯旭奔, E-mail: hxb@sdu.edu.cn;

方浩, Tel: 86-531-88381168, E-mail: haofangcn@sdu.edu.cn

Design, synthesis and anti-tumor activity evaluation of quinoline derivatives as histone deacetylase 8 inhibitors

Yi ZHOU¹, Wen-qing SHAO¹, Xin-ying YANG², Xu-ben HOU²^,^*, Hao FANG²^,^*

Affiliations

1. Department of Pharmacy, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Ji'nan 250021, China

2. Key Laboratory of Chemical Biology (Ministry of Education), School of Pharmaceutical Science, Cheeloo College of Medicine, Shandong University, Ji'nan 250012, China

出版时间: 2024-04-12 doi: 10.16438/j.0513-4870.2023-0523

文章导航

摘要

收起

作为I亚族组蛋白去乙酰化酶(histone deacetylase, HDAC) 成员, HDAC8是重要的抗肿瘤靶点。本研究基于课题组前期构建的HDAC8药效团模型, 以喹啉作为母核, 设计并合成了13个目标化合物, 其中SDFZ-E2和SDFZ-E3具有较好的HDAC8抑制活性和亚型选择性; 在细胞实验中, SDFZ-E2和SDFZ-E3表现出优于HDAC8选择性抑制剂PCI-34051的抗肿瘤细胞增殖活性; 还通过分子对接和分子动力学模拟研究了SDFZ-E2与HDAC8的结合模式。该项工作是以喹啉为母核发展HDAC8选择性抑制剂的一次探索, 相关活性化合物可以作为先导化合物进行进一步的结构改造。

关键词

喹啉衍生物 / 组蛋白去乙酰化酶8抑制剂 / 抗肿瘤活性 / 药效团

Abstract

收起

As a member of class I histone deacetylase (HDACs), HDAC8 is an important anticancer drug target. Based on our previously developed pharmacophore model for the HDAC8 inhibitor, we designed and synthesized 13 quinoline acid derivatives as new HDAC8 inhibitors. Among them, the compound SDFZ-E2 and SDFZ-E3 exhibited good HDAC8 inhibitory activities and isoform selectivity. In cell experiments, the target compounds SDFZ-E2 and SDFZ-E3 showed better antiproliferation activities than the known HDAC8 selective inhibitor PCI-34051. In addition, the proposed binding mode of SDFZ-E2 was investigated using molecular docking and molecular dynamics simulation. This work is a new attempt to develop HDAC8 selective inhibitor using quinoline as the scaffold, and the active compounds could serve as lead compounds for further structural optimization.

Key words

quinoline derivative / histone deacetylase 8 inhibitor / anti-tumor activity / pharmacophore model

引用本文

周易, 邵文晴, 杨新颖, 侯旭奔, 方浩. 喹啉类HDAC8选择性抑制剂的设计、合成及抗肿瘤活性研究. 药学学报, 2024 , 59 (4) : 979 -986 . DOI: 10.16438/j.0513-4870.2023-0523

Yi ZHOU, Wen-qing SHAO, Xin-ying YANG, Xu-ben HOU, Hao FANG. Design, synthesis and anti-tumor activity evaluation of quinoline derivatives as histone deacetylase 8 inhibitors[J]. Acta Pharmaceutica Sinica, 2024 , 59 (4) : 979 -986 . DOI: 10.16438/j.0513-4870.2023-0523

正文

收起

组蛋白去乙酰化酶(histone deacetylase, HDAC) 是一种具有催化N-乙酰化赖氨酸脱乙酰基功能的酶, 在表观遗传修饰中发挥重要作用^[1]。目前, 人体内已发现18种HDACs, 根据辅因子依赖性可分为两个家族: 锌离子依赖的HDACs (HDAC1~11) 和NAD⁺依赖的Sirtuin家族(SIRT1~7)。根据序列相似性, 锌离子依赖的HDACs可以分为I (HDAC1~3和HDAC8)、II (HDAC4~7和HDAC9~10)、IV (HDAC11) 三个亚族^{[2, 3]}。

作为I亚族HDACs成员, HDAC8由377个氨基酸(42 kDa)构成, 广泛分布于细胞核与细胞质中^[4]。除了组蛋白外, HDAC8可以催化多种非组蛋白的底物去乙酰化, 例如染色体结构维持蛋白3 (structural maintenance of chromosomes protein 3, SMC3)、雌激素相关受体α (estrogen related receptor alpha, ERRα)、p53等^[5-7], 因此参与了多种生命活动和信号转导。目前研究已经证实, HDAC8的异常表达与恶性肿瘤等多种疾病密切相关^[8-10]。近年来, 随着对HDAC8蛋白结构及生理功能研究的深入, HDAC8已成为重要的抗肿瘤药物靶标。目前已上市的非选择性HDAC抑制剂伏立诺他(vorinostat, SAHA)、贝利司他(belinostat, PXD-101)、帕比司他(panobinostat, LBH-589) 均对HDAC8具有较好的抑制活性, 但普遍缺乏选择性, 对其他HDAC亚型同样具有抑制作用^[11] (图 1)。近年来, HDAC8选择性抑制剂的研究成为抗肿瘤药物研发领域的重要方向, 部分HDAC8选择性抑制剂如PCI-34051 (图 1) 等被发现, 为HDAC8抑制剂的研究打下基础^[12]。

在前期工作中, 本课题组^[13]以HDAC8-抑制剂晶体结构复合物为训练集, 通过引入自定义的锌离子螯合基团(zinc binding group, ZBG) 元素, 构建了基于ZBG的HDAC8抑制剂药效团模型, 其主要特征是酶表面识别基团与锌离子螯合基团呈L型, 并以此为基础发现了HDAC8选择性抑制剂H8-A5 (图 1), 这为设计HDAC8抑制剂提供了理论基础。此外, 在课题组的前期研究中, 以喹啉为母核设计并合成了多个系列HDACs抑制剂, 部分化合物表现出了较好的HDACs抑制活性和抗肿瘤细胞增殖活性^{[14, 15]}。本研究以喹啉为母核, 通过在4位引入不同的取代基作为酶表面识别区, 设计一系列新型喹啉类HDAC8抑制剂, 代表性结构能够很好的匹配前期发展的HDAC8抑制剂药效团模型(图 2)。本研究共合成了13个目标化合物, 并通过核磁共振氢谱(hydrogen nuclear magnetic resonance spectroscopy, ¹H NMR)、核磁共振碳谱(carbon nuclear magnetic resonance spectroscopy, ¹³C NMR) 以及高分辨质谱(high resolution mass spectroscopy, HRMS)确证其化学结构。生物活性测试发现, 部分化合物具有较好的HDAC8抑制活性和亚型选择性, 并且对多个肿瘤细胞表现出了抗增殖活性。该项工作是以喹啉为母核发展HDAC8选择性抑制剂的一次探索, 相关活性化合物可以作为先导化合物进行进一步的结构改造。

结果与讨论

收起

1 化学合成

如路线1所示, 以苯胺为起始原料, 与丁炔二酸甲酯发生亲核加成反应后, 在多聚磷酸存在条件下成环得到中间体3。随后通过威廉姆森反应, 在4位酚羟基引入不同的侧链, 然后在碱性条件下脱除甲酯, 最后经缩合反应引入异羟肟酸获得目标化合物SDFZ-E1~SDFZ-E13 (表 1)。

2 生物活性测试

首先, 应用重组的人HDAC8蛋白对13个目标化合物进行了HDAC8抑酶活性测试, 选取已上市的HDAC泛抑制剂SAHA和HDAC8选择性抑制剂PCI-34051作为阳性对照。相关结果总结于表 2, 其中SDFZ-E2、SDFZ-E3、SDFZ-E12均表现出了微摩尔级的半数抑制浓度(half maximal inhibitory concentration, IC₅₀), 特别是SDFZ-E2的HDAC8抑制活性(IC₅₀ = 2.49 μmol·L^-1) 优于阳性对照药SAHA (IC₅₀ = 6.39 μmol·L^-1), 但是与PCI-34051 (IC₅₀ = 0.051 μmol·L^-1) 相比仍有差距。

随后, 对目标化合物SDFZ-E2、SDFZ-E3和SDFZ-E12进行了HDAC亚型选择性测试, 选取已上市的HDAC泛抑制剂SAHA作为阳性对照, 结果总结于表 3。目标化合物对HDAC1、HDAC2和HDAC6均表现出较差的抑制活性。其中, 目标化合物SDFZ-E2的HDAC8抑制活性较SAHA强2.6倍, 而HDAC1、HDAC2和HDAC6活性远弱于SAHA, 表明该目标化合物对HDAC8具有较好的亚型选择性, 符合设计目标。

根据以上结果, 目标化合物对HDAC8具有较好的抑制活性和亚型选择性, 进一步选取了三种高表达HDAC8的肿瘤细胞[人神经胶质瘤细胞SK-N-BE(2)、人弥漫性大B细胞淋巴瘤细胞SU-DHL-2和人急性T淋巴细胞白血病细胞Jurkat], 采用CCK-8法进行了抗细胞增殖实验^{[16, 17]}。结果见表 4, SDFZ-E2、SDFZ-E3、SDFZ-E12在SK-N-B(2) 和SU-DHL-2细胞中均表现出了微摩尔级的抗增殖活性, SDFZ-E2、SDFZ-E3在Jurkat细胞中, 也表现出了微摩尔级别的IC₅₀。与HDAC8选择性抑制剂PCI-34051相比, SDFZ-E2、SDFZ-E3在三种测试的细胞中均优于PCI-34051, 尤其在SU-DHL-2细胞中, SDFZ-E3 (IC₅₀ = 3.72 μmol·L^-1) 和SDFZ-E2 (IC₅₀ = 11.25 μmol·L^-1) 的抗增殖活性优于PCI-34051 (IC₅₀ = 21.12 μmol·L^-1)。

3 目标化合物与HDAC8的结合模式研究

为了进一步探究目标化合物和HDAC8的结合模式, 选取代表性化合物SDFZ-E2和HDAC8 (PDB ID: 5FCW) 晶体结构, 应用Schrodinger软件进行了分子对接和分子动力学模拟, 研究了SDFZ-E2与HDAC8的相互作用。

分子对接的结果表明, SDFZ-E2结构中的异羟肟酸部分深入了HDAC8催化口袋并与锌离子螯合, 喹啉环母核占据了疏水的催化口袋(图 3)。特别是, 喹啉4位的取代基作为酶表面识别基团, 与HDAC8表面由参与催化反应的Tyr306形成的特异性口袋结合(图 3)。该口袋位于L1 Loop和L6 Loop区之间, 为HDAC8所特有, 而在其他HDAC亚型中由于L1 Loop和L6 Loop结合紧密, 参与催化反应的Tyr306不外露。上述研究有助于理解SDFZ-E2具有较好的HDAC8选择性的结构基础。

进一步对SDFZ-E2和HDAC8的分子对接复合物进行了10 ns的分子动力学(molecular dynamics, MD) 模拟研究。如图 4A所示, 蛋白-小分子复合物在4 ns的MD后基本稳定, 而SDFZ-E2在10 ns的MD模拟过程中始终占据HDAC8特异口袋, 表明分子对接所预测的结合模式较为稳定。另一方面, 根据蛋白各个氨基酸在MD模拟中的RMSF值, 发现Ser13和Asp87表现出较大的柔性(RMSF > 3 Å), 但上述氨基酸位于远离催化口袋的Loop区, 与SDFZ-E2与HDAC8的结合关系不密切(图 4B)。在MD模拟中, SDFZ-E2结构中异羟肟酸的羟基和羰基与锌离子螯合较好, 锌离子与羰基O原子和羟基O原子的距离在模拟过程中始终小于3 Å, 符合形成配位键的距离(图 4C、D)。

4 小结

本文设计并合成了一系列新型喹啉类HDAC8小分子抑制剂。在化合物设计中, 根据前期发展的药效团模型, 以喹啉作为母核, 通过在4位引入不同的取代基探究目标化合物的构效关系。在活性评价中, 目标化合物SDFZ-E2表现出优于已上市HDAC抑制剂SAHA的HDAC8抑制活性和亚型选择性; 在抗肿瘤细胞增殖活性实验中, 目标化合物SDFZ-E2、SDFZ-E3表现出了较好的抗肿瘤活性, 显著优于HDAC8选择性抑制剂PCI-34051; 还通过分子对接和分子动力学模拟, 研究了代表性目标化合物和HDAC8的相互作用。该项工作是以喹啉为母核发展HDAC8选择性抑制剂的一次探索, 相关活性化合物可以作为先导化合物进行进一步的结构优化, 以获得活性更高、选择性更强的HDAC8抑制剂。

实验部分

收起

本实验所有试剂和溶剂未经说明的均为分析纯。HDAC蛋白(HDACs混酶HDAC1、HDAC2、HDAC6、HDAC8) 购自Enzo公司。HDAC底物购自上海毕得公司。Tris-base、胰酶购自北京索莱宝公司。HDACs抑制剂TSA购自MCE公司。本实验中所有¹H NMR谱和¹³C NMR谱通过Bruker DRX光谱仪测定(400 MHz)。HRMS谱由SCI-EX X500e Q-TOF LC/MS系统测定。本实验所有熔点用电热熔点仪测定。超纯水由MilliQ超纯水制备系统制得。

1 化学合成

1.1 2-(苯氨基)富马酸二甲酯(2)的合成

苯胺(20 g, 215 mmol) 溶于40 mL甲醇, 0 ℃下逐滴滴入丁炔二酸二甲酯(30.54 g, 215 mmol)。随后, 将反应液加热至80 ℃回流8 h。旋去溶剂, 得到黄色油状产物, 用硅胶扮样后, 柱层析纯化(石油醚∶乙酸乙酯= 99∶1), 最终得到纯的淡黄色油状中间体2, 产率70%。

1.2 4-羟基喹啉-2-羧酸甲酯(3)的合成

将30 g中间体2与100 g多聚磷酸用机械搅拌混合, 缓慢加热至120 ℃, 待反应液逐渐变红后保持温度2 h。随后, 向反应液中缓慢滴加10%的碳酸钾溶液至所有黏稠状液体溶解, 并不再产生气泡, 过滤悬浊液, 滤饼用少量水洗, 烘干即得中间体3, 收率75%。棕色粉末, 熔点: 224~226 ℃。

1.3 4-甲氧基喹啉-2-羧酸甲酯(4a)的合成

中间体3 (2 g, 10 mmol)、碘甲烷(1.68 g, 11 mmol)、碳酸钾(4.08 g, 30 mmol) 溶于50 mL DMF, 加热至70 ℃反应8 h。自然冷却至室温后, 加水淬灭反应, 静置沉淀, 过滤分离固体, 用少量水洗滤饼, 烘干即得4a。产率: 74%, 棕色粉末, 熔点: 224~226 ℃。

1.4 4-乙氧基喹啉-2-羧酸甲酯(4b)的合成

中间体3 (2 g, 10 mmol)、溴乙烷(1.09 g, 10 mmol)、碳酸钾(4.08 g, 30 mmol) 溶于50 mL DMF, 加热至70 ℃反应8 h。自然冷却至室温后, 加水淬灭反应, 乙酸乙酯萃取, 水和饱和食盐水各洗一次有机层, 无水硫酸镁干燥, 旋干得化合物4b。产率: 65%, 棕色油。

1.5 4-(苄氧基)喹啉-2-羧酸甲酯(4c) 的合成

合成与纯化方法类似4b。产率: 68%, 棕色油。

1.6 4-((4-溴苄基)氧基)喹啉-2-羧酸甲酯(4d)的合成

合成与纯化方法类似4a。产率: 70%, 白色固体, 熔点: 180~182 ℃。

1.7 4-((3-溴苄基)氧基)喹啉-2-羧酸甲酯(4e)的合成

合成与纯化方法类似4a。产率: 72%, 白色固体, 熔点: 176~178 ℃。

1.8 4-((4-(叔丁基)苄基)氧基)喹啉-2-羧酸甲酯(4f)的合成

合成与纯化方法类似4b。产率: 65%, 黑色油。

1.9 4-((4-氯苄基)氧基)喹啉-2-羧酸甲酯(4g)的合成

合成与纯化方法类似4a。产率: 68%, 白色固体, 熔点: 220~222 ℃。

1.10 4-(2-(4-氯-3, 5-二甲基苯氧基)乙氧基)喹啉-2-羧酸甲酯(4h)的合成

合成与纯化方法类似4b。产率: 55%, 棕色油。

1.11 4-((4-甲氧基苄基)氧基)喹啉-2-羧酸甲酯(4i)的合成

合成与纯化方法类似4a。产率: 75%, 白色固体, 熔点: 190~192 ℃。

1.12 4-苯乙氧基喹啉-2-羧酸甲酯(4j)的合成

合成与纯化方法类似4a。产率: 74%, 白色固体, 熔点: 170~172 ℃。

1.13 4-([1, 1′-联苯]-4-基甲氧基)喹啉-2-羧酸甲酯(4k)的合成

合成与纯化方法类似4a。产率: 58%, 白色固体, 熔点: > 300 ℃。

1.14 4-(萘-1-基甲氧基)喹啉-2-羧酸甲酯(4l)的合成

合成与纯化方法类似4b。产率: 60%, 黄色油。

1.15 4-(2-(萘-1-基氧基)乙氧基)喹啉-2-羧酸甲酯(4m)的合成

合成与纯化方法类似4a。产率: 60%, 淡黄色固体, 熔点: 194~196 ℃。

1.16 4-甲氧基喹啉-2-羧酸(5a)的合成

中间体4a (1.5 g, 6.91 mmol) 和氢氧化锂(0.83 g, 34.53 mmol) 溶于80 mL的四氢呋喃/水(2/1) 溶液, 室温搅拌12 h, 反应完毕后旋去四氢呋喃, 用2 mol·L^-1盐酸调节pH至中性, 过滤分离沉淀, 用少量水洗滤饼, 烘干, 乙酸乙酯打浆, 过滤, 烘干, 即得产物5a。产率: 85%, 白色固体, 熔点: > 300 ℃。

1.17 4-乙氧基喹啉-2-羧酸(5b)的合成

合成与纯化方法类似5a。产率: 80%, 黄白色固体, 熔点: 104~106 ℃。

1.18 4-(苄氧基)喹啉-2-羧酸(5c)的合成

合成与纯化方法类似5a。产率: 75%, 白色固体, 熔点: 272~274 ℃。

1.19 4-((4-溴苄基)氧基)喹啉-2-羧酸(5d)的合成

合成与纯化方法类似5a。产率: 72%, 白色固体, 熔点: > 300 ℃。

1.20 4-((3-溴苄基)氧基)喹啉-2-羧酸(5e)的合成

合成与纯化方法类似5a。产率: 78%, 白色固体, 熔点: > 300 ℃。

1.21 4-((4-(叔丁基)苄基)氧基)喹啉-2-羧酸(5f)的合成

合成与纯化方法类似5a。产率: 64%, 棕色固体, 熔点: 286~288 ℃。

1.22 4-((4-氯苄基)氧基)喹啉-2-羧酸(5g)的合成

合成与纯化方法类似5a。产率: 70%, 白色固体, 熔点: 254~256 ℃。

1.23 4-(2-(4-氯-3, 5-二甲基苯氧基)乙氧基)喹啉-2-羧酸(5h)的合成

合成与纯化方法类似5a。产率: 55%, 黄白色固体, 熔点: 196~198 ℃。

1.24 4-((4-甲氧基苄基)氧基)喹啉-2-羧酸(5i)的合成

合成与纯化方法类似5a。产率: 78%, 白色固体, 熔点: > 300 ℃。

1.25 4-苯乙基喹啉-2-羧酸(5j)的合成

合成与纯化方法类似5a。产率: 65%, 白色固体, 熔点: 236~238 ℃。

1.26 4-([1, 1′-联苯] -4-基甲氧基)喹啉-2-羧酸(5k)的合成

合成与纯化方法类似5a。产率: 60%, 白色固体, 熔点: > 300 ℃。

1.27 4-(萘-1-基甲氧基)喹啉-2-羧酸(5l)的合成

合成与纯化方法类似5a。产率: 80%, 白色固体, 熔点: > 300 ℃。

1.28 4-(2-(萘-1-基氧基)乙氧基)喹啉-2-羧酸(5m)的合成

合成与纯化方法类似5a。产率: 50%, 黄白色固体, 熔点: 160~162 ℃。

1.29 N-羟基-4-(萘-1-基甲氧基)喹啉-2-羧酰胺(SDFZ-E1)的合成

将5l(0.33 g, 1 mmol)、O-(四氢-2H-吡喃-2-基)羟胺(0.14 g, 1.2 mmol) 和HATU (0.45 g, 1.2 mmol) 在冰浴下溶解在DMF中, 加入NMM (0.3 mL, 3 mmol), 在氮气保护下搅拌8 h, 倒入水中, 并用乙酸乙酯(3×20 mL) 萃取。合并的有机层用5%柠檬酸, 饱和NaHCO₃和饱和食盐水洗涤。用无水MgSO₄干燥后, 过滤并旋干, 得到粗中间体。将该中间体溶于氯化氢的乙酸乙酯饱和溶液中。搅拌10 min后, 过滤沉淀, 得到粗产物。通过无水乙醇重结晶纯化最终得到终产物。

1.30 SDFZ-E2~SDFZ-E13的合成

SDFZ-E2~SDFZ-E13的合成与SDFZ-E1的合成类似。

2 生物活性评价

2.1 HDAC酶活测试

将10 μL酶溶液(HDAC1、HDAC2、HDAC6或HDAC8) 与不同浓度的受试化合物(50 μL) 混合。将混合物在37 ℃下孵育5 min, 然后加入荧光底物Boc-Lys(Ac)-AMC。在37 ℃下孵育30 min后, 通过添加100 μL含有胰蛋白酶和TSA的终止剂来停止反应, 并将去乙酰化的底物转化为具有荧光的化合物。20 min后, 使用微孔板读取器分别在390和460 nm的激发波长和发射波长下测量荧光强度。根据测试井相对于对照井的荧光强度读数计算抑制率, 并使用GraphPad Prism软件中的浓度/抑制数据的回归分析计算IC₅₀值^[18]。

2.2 抗肿瘤细胞增殖活性研究

本工作中的所有细胞都用RPMI-1640 (含有10%的胎牛血清) 培养。首先, 将持续培养至对数期的实体瘤细胞(3 000个细胞/孔) 或血液瘤细胞(10 000个细胞/孔) 接种在96孔板中。12 h后, 加入溶解在培养基中的不同浓度的药物。培养48 h后, 将CCK-8溶液加入每个孔中。将96孔板置于37 ℃孵育1~4 h, 待96孔板变为橘黄色后, 将96孔板置于摇床摇匀, 用酶标仪测量450 nm处的吸收值, 并计算不同浓度的抑制率和IC₅₀值。

3 结合模式预测

3.1 对接研究

应用Schrodinger 2018 suite中的LigPrep对目标化合物进行处理, 选取OPLS_2005全原子力场, 目标pH设为7 ± 2, 其余参数保持默认。随后, 导入人源的HDAC8蛋白结构(PDB ID: 5FCW), 应用Protein preparation wizard对其进行删除水分子、加氢、加电荷、调整锌离子附近组氨酸残基的质子化状态等预处理。接着, 以HDAC8催化位点为中心, 生成边长为20 Å的网格, 其余参数均保持默认。使用Schrodinger 2018 suite中的对接模块, 对处理好的配体和对接格子进行对接, 其余选项保持默认。选取打分最高的构象作为对接结果, 并用于后续的分子动力学模拟。

3.2 分子动力学模拟

应用Desmond进行^[19]对目标化合物-靶蛋白复合物进行分子动力学模拟, 以分子对接生成的复合物作为起始结构进行10 ns的分子动力学模拟。选用OPLS_2005全原子力场, SPC水模型, 引入Na⁺或Cl^-使体系电荷保持中性。结果的处理应用Schrodinger中的simulation events analysis模块, 分析蛋白和配体的RMSD变化, 蛋白中各氨基酸残基的RMSF变化, 关键原子间的距离变化。

作者贡献: 方浩对本文提供指导性支持, 设计论文总纲, 终审论文; 侯旭奔负责设计研究方案, 指导研究开展, 对文章进行修改; 杨新颖负责提供技术指导, 进行结构确证; 周易负责化学合成、生物评价、计算机模拟和文章写作; 邵文晴负责生物评价、计算机模拟和统计分析。

利益冲突: 本文不存在任何利益相关问题。

基金

收起

国家自然科学基金资助项目(81874288)
国家自然科学基金资助项目(82204198)

参考文献

收起

文献

收起

参考文献引证文献

排序方式：

[1]

Peixoto P, Cartron PF, Serandour AA, et al. From 1957 to nowadays: a brief history of epigenetics [J]. Int J Mol Sci, 2020, 21: 7571.

[2]

Witt O, Deubzer HE, Milde T, et al. HDAC family: what are the cancer relevant targets? [J]. Cancer Lett, 2009, 277: 8-21.

[3]

Gao X, Han W, Tian S, et al. Structurally novel HDAC inhibitors based on the trans-β-arylacryl tetrahydroisoquinoline scaffold: the design, synthesis, and anticancer activity [J]. Acta Pharm Sin (药学学报), 2023, 58: 413-422.

[4]

Kim JY, Cho H, Yoo J, et al. Pathological role of HDAC8: cancer and beyond [J]. Cells, 2022, 11: 3161.

[5]

Deardorff MA, Bando M, Nakato R, et al. HDAC8 mutations in Cornelia de Lange syndrome affect the cohesin acetylation cycle [J]. Nature, 2012, 489: 313-317.

[6]

Wilson BJ, Tremblay AM, Deblois G, et al. An acetylation switch modulates the transcriptional activity of estrogen-related receptor alpha [J]. Mol Endocrinol, 2010, 24: 1349-1358.

[7]

Chakrabarti A, Oehme I, Witt O, et al. HDAC8: a multifaceted target for therapeutic interventions [J]. Trends Pharmacol Sci, 2015, 36: 481-492.

[8]

Nakagawa M, Oda Y, Eguchi T, et al. Expression profile of class I histone deacetylases in human cancer tissues [J]. Oncol Rep, 2007, 18: 769-774.

[9]

Wu J, Du C, Lv Z, et al. The up-regulation of histone deacetylase 8 promotes proliferation and inhibits apoptosis in hepatocellular carcinoma [J]. Dig Dis Sci, 2013, 58: 3545-3553.

[10]

Vanaja GR, Ramulu HG, Kalle AM. Overexpressed HDAC8 in cervical cancer cells shows functional redundancy of tubulin deacetylation with HDAC6 [J]. Cell Commun Signal, 2018, 16: 20.

[11]

Liang T, Wang F, Elhassan RM, et al. Targeting histone deacetylases for cancer therapy: trends and challenges [J]. Acta Pharm Sin B, 2023, 13: 2425-2463.

[12]

Bai SY, Li ML, Ren Y, et al. HDAC8-inhibitor PCI-34051-induced exosomes inhibit human bronchial smooth muscle cell proliferation via miR-381-3p mediated TGFB3 [J]. Pulm Pharmacol Ther, 2021, 71: 102096.

[13]

Hou X, Du J, Liu R, et al. Enhancing the sensitivity of pharmacophore-based virtual screening by incorporating customized ZBG features: a case study using histone deacetylase 8 [J]. J Chem Inf Model, 2015, 55: 861-871.

[14]

Wang L, Hou XB, Fu HS, et al. Design, synthesis and preliminary bioactivity evaluations of substituted quinoline hydroxamic acid derivatives as novel histone deacetylase (HDAC) inhibitors [J]. Bioorg Med Chem, 2015, 23: 4364-4374.

[15]

Chen C, Hou X, Wang G, et al. Design, synthesis and biological evaluation of quinoline derivatives as HDAC class I inhibitors [J]. Eur J Med Chem, 2017, 133: 11-23.

[16]

Heimburg T, Kolbinger FR, Zeyen P, et al. Structure-based design and biological characterization of selective histone deacetylase 8 (HDAC8) inhibitors with anti-neuroblastoma activity [J]. J Med Chem, 2017, 60: 10188-10204.

[17]

Sixto-Lopez Y, Gomez-Vidal JA, De Pedro N, et al. Hydroxamic acid derivatives as HDAC1, HDAC6 and HDAC8 inhibitors with antiproliferative activity in cancer cell lines [J]. Sci Rep, 2020, 10: 10462.

[18]

Zhou Y, Liu X, Xue J, et al. Discovery of peptide boronate derivatives as histone deacetylase and proteasome dual inhibitors for overcoming bortezomib resistance of multiple myeloma [J]. J Med Chem, 2020, 63: 4701-4715.

[19]

Shaw DE. A fast, scalable method for the parallel evaluation of distance-limited pairwise particle interactions [J]. J Comput Chem, 2005, 26: 1318-1328.

2024年第59卷第4期

PDF下载

156

引用本文

BibTeX

文章信息

doi: 10.16438/j.0513-4870.2023-0523

接收时间：2023-06-28
首发时间：2025-11-28
出版时间：2024-04-12

补充材料

相关文章

文章信息

作者

出版历史

收稿日期：2023-06-28
修回日期：2023-11-28

基金

国家自然科学基金资助项目(81874288)

国家自然科学基金资助项目(82204198)

作者信息

1.山东第一医科大学附属省立医院药学部, 山东济南 250021

2.山东大学药学院, 天然产物化学生物学教育部重点实验室, 山东济南 250012

通讯作者:

^*侯旭奔, E-mail: hxb@sdu.edu.cn;

方浩, Tel: 86-531-88381168, E-mail: haofangcn@sdu.edu.cn

参考文献

分享链接

https://castjournals.cast.org.cn/joweb/yxxb/CN/10.16438/j.0513-4870.2023-0523

分享至

全文二维码

扫描看全文

引用本文

BibTeX

本文的引用情况

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

关闭全屏

BibTeX
EndNote
RefWorks
TxT

Compound	NMR spectrum and HRMS result	Property	Melting point/℃	Yield
SDFZ-E1	¹H NMR (400 MHz, DMSO-d₆ (dimethyl sulfoxide-d₆)) δ 11.73 (s, 1H), 8.26-8.19 (m, 1H), 8.18-8.08 (m, 2H), 8.02 (t, J = 6.6 Hz, 2H), 7.95 (s, 1H), 7.91-7.80 (m, 2H), 7.61 (p, J = 7.4 Hz, 4H), 6.01 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 133.8, 132.1, 131.6, 131.6, 129.7, 129.0, 128.2, 127.6, 127.1, 126.6, 125.9, 124.3, 122.3, 121.7, 100.4, 70.0. HRMS (ESI⁺): m/z calculated for C₂₁H₁₆N₂O₃ 345.123 4, found [M+H]⁺ 345.121 0.	White solid	250-252	15%
SDFZ-E2	¹H NMR (400 MHz, DMSO-d₆) δ 11.77 (s, 1H), 8.25 (d, J = 8.4 Hz, 1H), 8.13 (d, J = 8.5 Hz, 1H), 7.90 (t, J = 7.7 Hz, 1H), 7.76 (s, 1H), 7.71 (t, J = 7.7 Hz, 1H), 7.60 (d, J = 7.5 Hz, 2H), 7.52-7.36 (m, 3H), 5.55 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 136.3, 136.3, 131.4, 131.4, 129.8, 129.1, 128.7, 128.7, 128.1, 122.2, 70.8. HRMS (ESI⁺): m/z calculated for C₁₇H₁₄N₂O₃ 295.107 7, found [M+H]⁺ 295.105 2.	White solid	248-250	18%
SDFZ-E3	¹H NMR (400 MHz, DMSO-d₆) δ 11.48 (s, 1H), 9.20 (s, 1H), 8.23 (d, J = 8.3 Hz, 1H), 8.04 (d, J = 8.4 Hz, 1H), 7.84 (q, J = 6.9, 6.3 Hz, 1H), 7.66 (q, J = 6.6 Hz, 4H), 7.56 (d, J = 8.0 Hz, 2H), 5.49 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 163.2, 162.0, 151.9, 147.7, 139.2, 131.5, 131.3, 131.3, 131.1, 130.7, 129.5, 127.7, 127.6, 127.1, 127.1, 122.2, 122.1, 121.6, 99.5, 69.6. HRMS (ESI⁺): m/z calculated for C₁₇H₁₃BrN₂O₃ 373.018 2, found [M+H]⁺ 373.018 0.	Yellowish solid	220-222	22%
SDFZ-E4	¹H NMR (400 MHz, DMSO-d₆) δ 11.90 (s, 1H), 8.18 (dd, J = 23.3, 8.7 Hz, 2H), 8.04-7.63 (m, 4H), 7.59-7.51 (m, 2H), 7.07-6.99 (m, 2H), 5.49 (d, J = 5.3 Hz, 2H), 3.79 (s, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 159.9, 159.7, 140.6, 133.4, 130.8, 130.4, 129.6, 128.3, 124.6, 114.5, 114.4, 106.0, 100.5, 71.7, 71.2, 55.6, 55.5, 46.7. HRMS (ESI⁺): m/z calculated for C₁₈H₁₆N₂O₄ 325.118 3, found [M+H]⁺ 325.118 8.	White solid	248-250	16%
SDFZ-E5	¹H NMR (400 MHz, DMSO-d₆) δ 11.85 (s, 1H), 8.24 (d, J = 8.4 Hz, 1H), 8.16 (d, J = 8.5 Hz, 1H), 7.92 (t, J = 7.8 Hz, 1H), 7.82 (d, J = 3.4 Hz, 1H), 7.71 (t, J = 7.7 Hz, 1H), 7.57-7.45 (m, 4H), 5.52 (s, 2H), 1.31 (d, J = 1.9 Hz, 9H). ¹³C NMR (101 MHz, DMSO-d₆) δ 151.4, 132.8, 128.4, 125.9, 122.6, 121.5, 100.4, 71.7, 34.8, 31.5. HRMS (ESI⁺): m/z calculated for C₂₁H₂₂N₂O₃ 351.170 3, found [M+H]⁺ 351.167 7.	White solid	256-258	32%
SDFZ-E6	¹H NMR (400 MHz, DMSO-d₆) δ 11.58 (s, 1H), 8.24 (d, J = 8.4 Hz, 1H), 8.10 (d, J = 9.5 Hz, 1H), 7.88 (t, J = 7.4 Hz, 1H), 7.82-7.40 (m, 6H), 5.54 (t, J = 4.4 Hz, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 163.2, 162.1, 151.9, 147.7, 136.0, 132.0, 131.1, 130.2, 129.5, 127.6, 122.1, 121.8, 121.7, 99.6, 69.7. HRMS (ESI⁺): m/z calculated for C₁₇H₁₃BrN₂O₃ 373.018 2, found [M+H]⁺ 373.017 2.	White solid	286-288	16%
SDFZ-E7	¹H NMR (400 MHz, DMSO-d₆) δ 11.67 (s, 1H), 8.25 (d, J = 8.4 Hz, 1H), 8.11 (dd, J = 8.5, 3.4 Hz, 1H), 7.89 (t, J = 7.9 Hz, 1H), 7.84-7.63 (m, 4H), 7.57 (d, J = 8.1 Hz, 2H), 5.53 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 135.6, 132.1, 132.0, 131.8, 130.4, 128.2, 122.4, 122.0, 121.6, 100.2, 70.4, 57.8. HRMS (ESI⁺): m/z calculated for C₁₇H₁₃ClN₂O₃ 329.068 8, found [M+H]⁺ 329.067 4.	White solid	214-216	18%
SDFZ-E8	¹H NMR (400 MHz, DMSO-d₆) δ 11.79 (s, 1H), 8.26 (d, J = 9.2 Hz, 2H), 7.96 (s, 1H), 7.76 (s, 2H), 4.70-4.42 (m, 2H), 1.53 (dt, J = 9.2, 4.7 Hz, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 166.1, 158.8, 149.0, 143.0, 133.5, 128.7, 125.2, 122.7, 121.3, 100.4, 67.1, 14.6. HRMS (ESI⁺): m/z calculated for C₁₂H₁₂N₂O₃ 233.092 1, found [M+H]⁺ 233.091 0.	White solid	194-196	23%
SDFZ-E9	¹H NMR (400 MHz, DMSO-d₆) δ 11.65 (s, 1H), 8.25-8.02 (m, 3H), 7.87 (d, J = 8.4 Hz, 2H), 7.77 (s, 1H), 7.64 (t, J = 7.6 Hz, 1H), 7.46 (dq, J = 24.6, 7.8 Hz, 4H), 7.13 (d, J = 7.5 Hz, 1H), 4.91 (d, J = 4.9 Hz, 2H), 4.71 (d, J = 3.6 Hz, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 154.2, 134.5, 127.9, 126.9, 126.6, 125.8, 125.4, 122.3, 121.9, 121.6, 120.8, 106.3, 99.9, 68.7, 67.1. HRMS (ESI⁺): m/z calculated for C₂₂H₁₈N₂O₄ 375.133 9, found [M+H]⁺ 375.131 8.	Yellowish solid	192-194	14%
SDFZ-E10	¹H NMR (400 MHz, DMSO-d₆) δ 11.69 (s, 1H), 8.21 (d, J = 8.5 Hz, 1H), 8.10 (d, J = 8.5 Hz, 1H), 7.93-7.85 (m, 1H), 7.70 (t, J = 7.7 Hz, 1H), 7.61 (s, 1H), 4.17 (s, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 145.4, 144.9, 134.2, 133.6, 132.8, 132.2, 131.9, 5.3. HRMS (ESI⁺): m/z calculated for C₁₁H₁₀N₂O₃ 219.076 4, found [M+H]⁺ 219.075 2.	White solid	238-240	20%
SDFZ-E11	¹H NMR (400 MHz, DMSO-d₆) δ 10.92 (s, 1H), 8.34 – 8.20 (m, 3H), 7.93 (dd, J = 8.8, 6.7 Hz, 2H), 7.74 (h, J = 8.2 Hz, 5H), 7.55-7.32 (m, 4H), 4.19 (s, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 131.9, 131.4, 128.9, 128.4, 128.2, 128.1, 127.5, 127.2, 125.2, 124.4, 122.2, 121.8, 99.0, 57.3, 57.1. HRMS (ESI⁺): m/z calculated for C₂₃H₁₈N₂O₃ 371.139 0, found [M+H]⁺ 371.139 4.	White solid	> 300	13%
SDFZ-E12	¹H NMR (400 MHz, DMSO-d₆) δ 11.95 (s, 1H), 8.19 (dt, J = 8.4, 2.2 Hz, 2H), 7.99-7.90 (m, 1H), 7.83-7.71 (m, 2H), 7.44 (d, J = 7.1 Hz, 2H), 7.34 (t, J = 7.6 Hz, 2H), 7.28-7.22 (m, 1H), 4.68 (t, J = 6.4 Hz, 2H), 3.26 (t, J = 6.4 Hz, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 165.6, 159.1, 149.4, 138.3, 133.3, 129.5, 128.8, 128.7, 126.9, 125.7, 122.5, 121.3, 100.4, 71.3, 34.8. HRMS (ESI⁺): m/z calculated for C₁₈H₁₆N₂O₃ 309.123 4, found [M+H]⁺ 309.121 9.	White solid	180-182	21%
SDFZ-E13	¹H NMR (400 MHz, DMSO-d₆) δ 11.73 (s, 1H), 8.21-8.04 (m, 2H), 7.89 (t, J = 7.4 Hz, 1H), 7.69 (d, J = 6.9 Hz, 2H), 6.91 (s, 2H), 4.86-4.67 (m, 2H), 4.62-4.46 (m, 2H), 2.30 (s, 6H). ¹³C NMR (101 MHz, DMSO-d₆) δ 156.8, 137.1, 128.2, 125.8, 122.4, 121.4, 115.4, 100.0, 69.1, 66.5, 20.8. HRMS (ESI⁺): m/z calculated for C₂₀H₁₉ClN₂O₄ 387.110 6, found [M+H]⁺ 387.109 5.	White solid	216-218	17%

Compound

NMR spectrum and HRMS result

Property

Melting point/℃

Yield

SDFZ-E1

White solid

250-252

15%

SDFZ-E2

White solid

248-250

18%

SDFZ-E3

Yellowish solid

220-222

22%

SDFZ-E4

White solid

248-250

16%

SDFZ-E5

White solid

256-258

32%

SDFZ-E6

White solid

286-288

16%

SDFZ-E7

White solid

214-216

18%

SDFZ-E8

White solid

194-196

23%

SDFZ-E9

Yellowish solid

192-194

14%

SDFZ-E10

White solid

238-240

20%

SDFZ-E11

White solid

> 300

13%

SDFZ-E12

White solid

180-182

21%

SDFZ-E13

White solid

216-218

17%


SDFZ-E1	-1-Napthene	> 50
SDFZ-E2	-Ph	2.49 ± 0.49
SDFZ-E3	-3-Br-Ph	7.45 ± 0.26
SDFZ-E4	-4-Methoxy-Ph	32.98 ± 0.29
SDFZ-E5	-4-t-Bu-Ph	> 50
SDFZ-E6	-4-Br-Ph	> 50
SDFZ-E7	-4-Cl-Ph	> 50
SDFZ-E8	-CH₃	> 50
SDFZ-E9		> 50
SDFZ-E10	H	> 50
SDFZ-E11	-4-Ph-Ph	> 50
SDFZ-E12	-Bn	12.94 ± 1.14
SDFZ-E13		> 50
SAHA	-	6.39 ± 0.62
PCI-34051	-	0.051 ± 0.016

Compound

^aIC₅₀ of HDAC8/μmol·L^-1

SDFZ-E1

-1-Napthene

> 50

SDFZ-E2

-Ph

2.49 ± 0.49

SDFZ-E3

-3-Br-Ph

7.45 ± 0.26

SDFZ-E4

-4-Methoxy-Ph

32.98 ± 0.29

SDFZ-E5

-4-t-Bu-Ph

> 50

SDFZ-E6

-4-Br-Ph

> 50

SDFZ-E7

-4-Cl-Ph

> 50

SDFZ-E8

-CH₃

> 50

SDFZ-E9

> 50

SDFZ-E10

> 50

SDFZ-E11

-4-Ph-Ph

> 50

SDFZ-E12

-Bn

12.94 ± 1.14

SDFZ-E13

> 50

SAHA

6.39 ± 0.62

PCI-34051

0.051 ± 0.016

Compound	^aIC₅₀/μmol·L^-1
SDFZ-E2	> 50	> 50	33.12 ± 10.51	2.49 ± 0.49
SDFZ-E3	> 50	> 50	> 50	7.45 ± 0.26
SDFZ-E12	> 50	> 50	41.23 ± 3.21	12.95 ± 1.14
SAHA	0.037 ± 0.007	0.015 ± 0.002	0.090 ± 0.007	6.39 ± 0.62

Compound

^aIC₅₀/μmol·L^-1

HDAC1

HDAC2

HDAC6

HDAC8

SDFZ-E2

> 50

33.12 ± 10.51

2.49 ± 0.49

SDFZ-E3

> 50

7.45 ± 0.26

SDFZ-E12

> 50

41.23 ± 3.21

12.95 ± 1.14

SAHA

0.037 ± 0.007

0.015 ± 0.002

0.090 ± 0.007

6.39 ± 0.62

Compound	^aIC₅₀/μmol·L^-1
SDFZ-E2	13.99 ± 1.75	11.25 ± 0.94	42.07 ± 5.17
SDFZ-E3	9.68 ± 0.24	3.72 ± 1.12	10.65 ± 0.25
SDFZ-E12	37.44 ± 4.02	26.44 ± 5.90	> 200
PCI-34051	15.91 ± 0.58	21.12 ± 2.25	67.10 ± 9.97

Compound

^aIC₅₀/μmol·L^-1

SK-N-BE(2)

SU-DHL-2

Jurkat

SDFZ-E2

13.99 ± 1.75

11.25 ± 0.94

42.07 ± 5.17

SDFZ-E3

9.68 ± 0.24

3.72 ± 1.12

10.65 ± 0.25

SDFZ-E12

37.44 ± 4.02

26.44 ± 5.90

> 200

PCI-34051

15.91 ± 0.58

21.12 ± 2.25

67.10 ± 9.97