药学学报

Compd.	¹H NMR (400 MHz, DMSO-d₆)	ESI-MS (m/z) [M+H]⁺
K1	11.16 (s, 1H), 9.69 (s, 1H), 8.51 (d, J = 8.4 Hz, 1H), 7.84 (t, J = 7.9 Hz, 1H), 7.61 (d, J = 7.2 Hz, 1H), 5.26-5.12 (m, 2H), 3.94 (t, J = 9.1 Hz, 1H), 2.77 (d, J = 9.1 Hz, 1H), 2.59 (m, 4H), 2.47 (d, J = 7.0 Hz, 2H), 2.25-1.56 (m, 17H), 1.32 (d, J = 15.5 Hz, 2H), 1.25 (d, J = 10.2 Hz, 6H), 1.20 (s, 3H), 1.12 (m, 2H)	675.46
K2	11.15 (s, 1H), 9.70 (s, 1H), 8.48 (d, J = 8.4 Hz, 1H), 7.87-7.81 (m, 1H), 7.62 (d, J = 7.1 Hz, 1H), 5.22-5.07 (m, 2H), 3.96 (t, J = 9.1 Hz, 1H), 2.78 (d, J = 9.1 Hz, 1H), 2.61 (m, 5H), 2.43-1.92 (m, 17H), 1.63 (d, J = 9.3 Hz, 6H), 1.55-1.48 (m, 3H), 1.24 (s, 3H), 1.20 (s, 2H), 0.89-0.81 (m, 2H)	689.57
K3	11.15 (s, 1H), 9.70 (s, 1H), 8.48 (d, J = 8.4 Hz, 1H), 7.92-7.80 (m, 1H), 7.62 (d, J = 7.2 Hz, 1H), 5.26-5.11 (m, 2H), 3.96 (t, J = 9.1 Hz, 1H), 2.96-2.85 (m, 1H), 2.78 (d, J = 9.1 Hz, 1H), 2.60 (m, 6H), 2.48-1.91 (m, 17H), 1.74-1.56 (m, 6H), 1.48 (s, 2H), 1.33 (m, 3H), 1.20 (s, 3H), 1.11 (m, 2H)	703.64
K4	11.16 (s, 1H), 9.69 (s, 1H), 8.48 (d, J = 8.4 Hz, 1H), 7.89-7.78 (m, 1H), 7.62 (d, J = 7.0 Hz, 1H), 5.24-5.10 (m, 2H), 3.96 (t, J = 9.1 Hz, 1H), 2.79 (d, J = 9.1 Hz, 1H), 2.67-2.58 (m, 4H), 2.49-2.44 (m, 4H), 2.42-1.91 (m, 14H), 1.69-1.23 (m, 19H), 1.20 (s, 3H)	732.60
K5	11.15 (s, 1H), 9.70 (s, 1H), 8.50 (d, J = 8.4 Hz, 1H), 7.90-7.79 (m, 1H), 7.62 (d, J = 7.3 Hz, 1H), 6.03 (d, J = 3.4 Hz, 1H), 5.65-5.53 (m, 2H), 5.15 (m, 1H), 4.14 (t, J = 9.4 Hz, 1H), 3.18 (s, 1H), 2.96-2.83 (m, 2H), 2.61 (m, 2H), 2.48-1.57 (m, 19H), 1.50 (s, 2H), 1.43-1.29 (m, 1H), 1.28-1.21 (m, 2H), 1.16-0.98 (m, 2H)	687.30
K6	11.16 (s, 1H), 9.70 (s, 1H), 8.48 (t, J = 6.9 Hz, 1H), 7.84 (t, J = 7.9 Hz, 1H), 7.62 (d, J = 7.2 Hz, 1H), 6.68 (d, J = 8.3 Hz, 1H), 6.03 (d, J = 3.4 Hz, 1H), 5.57 (d, J = 3.0 Hz, 1H), 5.15 (m, 1H), 4.15 (t, J = 9.4 Hz, 1H), 2.37-1.98 (m, 14H), 1.64 (d, J = 7.9 Hz, 4H), 1.32 (d, J = 14.5 Hz, 6H), 1.25 (t, J = 6.6 Hz, 11H)	715.30
K7	11.10 (s, 1H), 8.09 (t, J = 5.7 Hz, 1H), 7.64-7.52 (m, 1H), 7.08-6.99 (m, 2H), 6.66 (t, J = 6.0 Hz, 1H), 5.98 (t, J = 5.6 Hz, 2H), 5.54 (d, J = 3.0 Hz, 1H), 4.17 (m, 3H), 3.14 (m, 2H), 2.65-2.54 (m, 2H), 2.15 (m, 7H), 1.74-1.62 (m, 4H), 1.49 (s, 3H), 1.25 (m, 4H)	591.20
K8	11.09 (s, 1H), 8.02 (s, 1H), 7.65-7.52 (m, 1H), 7.06 (m, 2H), 6.51 (t, J = 5.6 Hz, 1H), 6.02 (d, J = 3.4 Hz, 1H), 5.97-5.87 (m, 1H), 5.56 (d, J = 2.4 Hz, 1H), 5.05 (m, 1H), 4.11 (t, J = 9.4 Hz, 1H), 3.27-3.15 (m, 3H), 2.95-2.82 (m, 1H), 2.65-2.53 (m, 2H), 2.36-1.53 (m, 10H), 1.48 (s, 3H), 1.45-1.23 (m, 8H)	647.50
K9	11.09 (s, 1H), 7.59 (m, 1H), 7.06 (m, 2H), 6.02 (d, J = 3.4 Hz, 1H), 5.92 (t, J = 7.9 Hz, 1H), 5.54 (d, J = 3.1 Hz, 1H), 5.05 (m, 1H), 4.10 (t, J = 9.4 Hz, 1H), 3.30-2.56 (m, 12H), 2.35-1.52 (m, 12H), 1.48 (s, 3H), 1.24 (s, 3H), 1.03 (t, J = 12.6 Hz, 1H), 0.88-0.80 (m, 2H)	646.74
K10	11.09 (s, 1H), 8.14-7.83 (m, 1H), 7.58 (m, 2H), 7.06 (m, 3H), 6.51 (d, J = 5.3 Hz, 1H), 6.14-5.80 (m, 3H), 5.76 (s, 1H), 5.53 (d, J = 3.1 Hz, 2H), 5.03 (m, 2H), 4.05 (m, 2H), 3.31-3.25 (m, 3H), 3.24-3.11 (m, 2H), 3.03-2.91 (m, 2H), 2.91-2.81 (m, 1H), 2.80-2.65 (m, 2H), 2.65-2.54 (m, 3H), 2.32 (d, J = 12.1 Hz, 2H), 2.24 (m, 3H), 2.22-2.14 (m, 2H), 2.10 (m, 2H), 2.04 (m, 1H), 1.59 (t, J = 15.4 Hz, 5H), 1.48 (s, 4H), 0.84 (m, 2H)	716.75
K11	11.10 (s, 1H), 8.12 (s, 1H), 7.60 (m, 1H), 7.09 (m, 2H), 6.58 (t, J = 5.4 Hz, 1H), 6.01 (m, 1H), 5.95 (s, 1H), 5.57 (m, 1H), 5.06 (m, 1H), 4.09 (t, J = 9.1 Hz, 1H), 3.57 (d, J = 5.2 Hz, 2H), 3.44 (m, 3H), 3.30-2.53 (m, 8H), 2.14 (m, 7H), 1.48 (s, 3H), 1.25 (d, J = 5.5 Hz, 1H), 1.00 (m, 1H)	621.30
K12	11.09 (s, 1H), 8.11 (s, 1H), 7.59 (m, 1H), 7.15 (d, J = 8.6 Hz, 1H), 7.05 (d, J = 7.0 Hz, 1H), 6.02 (d, J = 3.4 Hz, 1H), 5.95 (t, J = 8.4 Hz, 1H), 5.57 (d, J = 3.1 Hz, 1H), 5.06 (m, 1H), 4.08 (t, J = 9.4 Hz, 1H), 3.60 (t, J = 5.4 Hz, 2H), 3.54-3.38 (m, 9H), 3.22-2.54 (m, 7H), 2.37-2.00 (m, 8H), 1.47 (s, 3H), 1.02 (t, J = 11.8 Hz, 1H)	664.51
K13	11.09 (s, 1H), 8.12 (t, J = 5.6 Hz, 1H), 7.59 (m, 1H), 7.15 (d, J = 8.6 Hz, 1H), 7.05 (d, J = 7.0 Hz, 1H), 6.02 (d, J = 3.4 Hz, 1H), 5.95 (t, J = 8.0 Hz, 1H), 5.76 (s, 1H), 5.57 (d, J = 3.1 Hz, 1H), 5.06 (m, 1H), 4.08 (t, J = 9.4 Hz, 1H), 3.65-3.36 (m, 15H), 2.77-2.57 (m, 3H), 2.37-1.52 (m, 10H), 1.47 (s, 3H), 1.28-1.21 (m, 2H)	709.40
K14	11.09 (s, 1H), 8.13 (t, J = 5.5 Hz, 1H), 7.59 (m, 1H), 7.15 (d, J = 8.6 Hz, 1H), 7.05 (d, J = 7.0 Hz, 1H), 6.04-5.89 (m, 2H), 5.57 (d, J = 3.1 Hz, 1H), 5.08-5.00 (m, 1H), 4.08 (t, J = 9.4 Hz, 1H), 3.63 (t, J = 5.4 Hz, 2H), 3.57-3.46 (m, 10H), 3.44 (s, 3H), 3.23-2.56 (m, 8H), 2.37-1.99 (m, 10H), 1.47 (s, 3H), 1.25 (d, J = 9.5 Hz, 2H)	752.49
K15	11.07 (s, 1H), 7.57 (m, 1H), 7.13 (t, J = 8.3 Hz, 2H), 6.03 (d, J = 3.5 Hz, 1H), 5.67-5.56 (m, 2H), 5.06 (m, 1H), 4.14 (t, J = 9.4 Hz, 1H), 3.55 (m, 6H), 2.88 (m, 3H), 2.74 (s, 1H), 2.34 (m, 3H), 2.22-1.52 (m, 11H), 1.50 (s, 3H), 1.42-1.01 (m, 4H)	656.56
K16	11.10 (s, 1H), 7.59 (m, 1H), 7.04 (m, 2H), 6.49 (d, J = 6.3 Hz, 1H), 6.02 (d, J = 3.5 Hz, 1H), 5.59 (m, 2H), 5.06 (m, 1H), 4.14 (t, J = 9.4 Hz, 2H), 2.96-2.82 (m, 2H), 2.69-2.55 (m, 2H), 2.35 (m, 6H), 2.23-1.96 (m, 5H), 1.76 (t, J = 9.3 Hz, 3H), 1.63 (m, 4H), 1.50 (s, 2H), 1.39 (d, J = 7.4 Hz, 1H), 1.27-1.23 (m, 3H)	656.73
K17	11.08 (s, 1H), 7.68 (m, 1H), 7.33 (m, 2H), 6.03 (d, J = 2.4 Hz, 1H), 5.65-5.51 (m, 2H), 5.09 (m, 1H), 4.14 (t, J = 9.3 Hz, 1H), 3.74 (d, J = 11.6 Hz, 2H), 2.87-2.81 (m, 3H), 2.66-2.54 (m, 3H), 2.44-1.72 (m, 13H), 1.51 (s, 3H), 1.44-1.36 (m, 3H), 1.24 (s, 3H), 1.13-1.00 (m, 3H)	684.59
K18	11.10 (s, 1H), 8.00 (t, J = 5.6 Hz, 1H), 7.81 (m, 1H), 7.48 (m, 2H), 6.02 (d, J = 3.4 Hz, 1H), 5.92 (t, J = 8.0 Hz, 1H), 5.54 (d, J = 2.9 Hz, 1H), 5.09 (m, 1H), 4.20 (t, J = 6.4 Hz, 2H), 4.10 (t, J = 9.4 Hz, 1H), 3.22 (m, 1H), 3.16 (d, J = 9.6 Hz, 1H), 3.01-2.56 (m, 5H), 2.55-2.52 (m, 1H), 2.34-2.00 (m, 7H), 1.79-1.72 (m, 2H), 1.58 (t, J = 11.3 Hz, 3H), 1.45-1.26 (m, 9H), 1.02 (m, 2H)	662.40
K19	11.06 (s, 1H), 8.00 (t, J = 5.7 Hz, 1H), 7.56 (d, J = 8.4 Hz, 1H), 7.08 (t, J = 5.3 Hz, 1H), 6.95 (d, J = 1.8 Hz, 1H), 6.85 (m, 1H), 6.03 (d, J = 3.5 Hz, 1H), 5.92 (t, J = 7.9 Hz, 1H), 5.54 (d, J = 3.1 Hz, 1H), 5.03 (m, 1H), 4.10 (t, J = 9.4 Hz, 1H), 3.26-3.12 (m, 4H), 3.02-2.52 (m, 6H), 2.37-1.51 (m, 9H), 1.48 (s, 3H), 1.34 (m, 6H), 1.24 (s, 3H)	647.50
K20	11.06 (s, 1H), 7.99 (t, J = 5.7 Hz, 1H), 7.53 (m, 1H), 7.09 (t, J = 5.2 Hz, 1H), 6.95 (d, J = 1.7 Hz, 1H), 6.85 (m, 1H), 6.02 (d, J = 3.5 Hz, 1H), 5.92 (t, J = 8.0 Hz, 1H), 5.54 (d, J = 3.1 Hz, 1H), 5.03 (m, 1H), 4.10 (t, J = 9.4 Hz, 1H), 3.25-2.52 (m, 14H), 2.37-1.96 (m, 8H), 1.64-1.53 (m, 3H), 1.24 (s, 3H), 1.21-1.16 (m, 3H), 1.02 (m, 2H)	661.80

Compd.	¹H NMR (400 MHz, DMSO-d₆)	ESI-MS (m/z) [M+H]⁺
K1	11.16 (s, 1H), 9.69 (s, 1H), 8.51 (d, J = 8.4 Hz, 1H), 7.84 (t, J = 7.9 Hz, 1H), 7.61 (d, J = 7.2 Hz, 1H), 5.26-5.12 (m, 2H), 3.94 (t, J = 9.1 Hz, 1H), 2.77 (d, J = 9.1 Hz, 1H), 2.59 (m, 4H), 2.47 (d, J = 7.0 Hz, 2H), 2.25-1.56 (m, 17H), 1.32 (d, J = 15.5 Hz, 2H), 1.25 (d, J = 10.2 Hz, 6H), 1.20 (s, 3H), 1.12 (m, 2H)	675.46
K2	11.15 (s, 1H), 9.70 (s, 1H), 8.48 (d, J = 8.4 Hz, 1H), 7.87-7.81 (m, 1H), 7.62 (d, J = 7.1 Hz, 1H), 5.22-5.07 (m, 2H), 3.96 (t, J = 9.1 Hz, 1H), 2.78 (d, J = 9.1 Hz, 1H), 2.61 (m, 5H), 2.43-1.92 (m, 17H), 1.63 (d, J = 9.3 Hz, 6H), 1.55-1.48 (m, 3H), 1.24 (s, 3H), 1.20 (s, 2H), 0.89-0.81 (m, 2H)	689.57
K3	11.15 (s, 1H), 9.70 (s, 1H), 8.48 (d, J = 8.4 Hz, 1H), 7.92-7.80 (m, 1H), 7.62 (d, J = 7.2 Hz, 1H), 5.26-5.11 (m, 2H), 3.96 (t, J = 9.1 Hz, 1H), 2.96-2.85 (m, 1H), 2.78 (d, J = 9.1 Hz, 1H), 2.60 (m, 6H), 2.48-1.91 (m, 17H), 1.74-1.56 (m, 6H), 1.48 (s, 2H), 1.33 (m, 3H), 1.20 (s, 3H), 1.11 (m, 2H)	703.64
K4	11.16 (s, 1H), 9.69 (s, 1H), 8.48 (d, J = 8.4 Hz, 1H), 7.89-7.78 (m, 1H), 7.62 (d, J = 7.0 Hz, 1H), 5.24-5.10 (m, 2H), 3.96 (t, J = 9.1 Hz, 1H), 2.79 (d, J = 9.1 Hz, 1H), 2.67-2.58 (m, 4H), 2.49-2.44 (m, 4H), 2.42-1.91 (m, 14H), 1.69-1.23 (m, 19H), 1.20 (s, 3H)	732.60
K5	11.15 (s, 1H), 9.70 (s, 1H), 8.50 (d, J = 8.4 Hz, 1H), 7.90-7.79 (m, 1H), 7.62 (d, J = 7.3 Hz, 1H), 6.03 (d, J = 3.4 Hz, 1H), 5.65-5.53 (m, 2H), 5.15 (m, 1H), 4.14 (t, J = 9.4 Hz, 1H), 3.18 (s, 1H), 2.96-2.83 (m, 2H), 2.61 (m, 2H), 2.48-1.57 (m, 19H), 1.50 (s, 2H), 1.43-1.29 (m, 1H), 1.28-1.21 (m, 2H), 1.16-0.98 (m, 2H)	687.30
K6	11.16 (s, 1H), 9.70 (s, 1H), 8.48 (t, J = 6.9 Hz, 1H), 7.84 (t, J = 7.9 Hz, 1H), 7.62 (d, J = 7.2 Hz, 1H), 6.68 (d, J = 8.3 Hz, 1H), 6.03 (d, J = 3.4 Hz, 1H), 5.57 (d, J = 3.0 Hz, 1H), 5.15 (m, 1H), 4.15 (t, J = 9.4 Hz, 1H), 2.37-1.98 (m, 14H), 1.64 (d, J = 7.9 Hz, 4H), 1.32 (d, J = 14.5 Hz, 6H), 1.25 (t, J = 6.6 Hz, 11H)	715.30
K7	11.10 (s, 1H), 8.09 (t, J = 5.7 Hz, 1H), 7.64-7.52 (m, 1H), 7.08-6.99 (m, 2H), 6.66 (t, J = 6.0 Hz, 1H), 5.98 (t, J = 5.6 Hz, 2H), 5.54 (d, J = 3.0 Hz, 1H), 4.17 (m, 3H), 3.14 (m, 2H), 2.65-2.54 (m, 2H), 2.15 (m, 7H), 1.74-1.62 (m, 4H), 1.49 (s, 3H), 1.25 (m, 4H)	591.20
K8	11.09 (s, 1H), 8.02 (s, 1H), 7.65-7.52 (m, 1H), 7.06 (m, 2H), 6.51 (t, J = 5.6 Hz, 1H), 6.02 (d, J = 3.4 Hz, 1H), 5.97-5.87 (m, 1H), 5.56 (d, J = 2.4 Hz, 1H), 5.05 (m, 1H), 4.11 (t, J = 9.4 Hz, 1H), 3.27-3.15 (m, 3H), 2.95-2.82 (m, 1H), 2.65-2.53 (m, 2H), 2.36-1.53 (m, 10H), 1.48 (s, 3H), 1.45-1.23 (m, 8H)	647.50
K9	11.09 (s, 1H), 7.59 (m, 1H), 7.06 (m, 2H), 6.02 (d, J = 3.4 Hz, 1H), 5.92 (t, J = 7.9 Hz, 1H), 5.54 (d, J = 3.1 Hz, 1H), 5.05 (m, 1H), 4.10 (t, J = 9.4 Hz, 1H), 3.30-2.56 (m, 12H), 2.35-1.52 (m, 12H), 1.48 (s, 3H), 1.24 (s, 3H), 1.03 (t, J = 12.6 Hz, 1H), 0.88-0.80 (m, 2H)	646.74
K10	11.09 (s, 1H), 8.14-7.83 (m, 1H), 7.58 (m, 2H), 7.06 (m, 3H), 6.51 (d, J = 5.3 Hz, 1H), 6.14-5.80 (m, 3H), 5.76 (s, 1H), 5.53 (d, J = 3.1 Hz, 2H), 5.03 (m, 2H), 4.05 (m, 2H), 3.31-3.25 (m, 3H), 3.24-3.11 (m, 2H), 3.03-2.91 (m, 2H), 2.91-2.81 (m, 1H), 2.80-2.65 (m, 2H), 2.65-2.54 (m, 3H), 2.32 (d, J = 12.1 Hz, 2H), 2.24 (m, 3H), 2.22-2.14 (m, 2H), 2.10 (m, 2H), 2.04 (m, 1H), 1.59 (t, J = 15.4 Hz, 5H), 1.48 (s, 4H), 0.84 (m, 2H)	716.75
K11	11.10 (s, 1H), 8.12 (s, 1H), 7.60 (m, 1H), 7.09 (m, 2H), 6.58 (t, J = 5.4 Hz, 1H), 6.01 (m, 1H), 5.95 (s, 1H), 5.57 (m, 1H), 5.06 (m, 1H), 4.09 (t, J = 9.1 Hz, 1H), 3.57 (d, J = 5.2 Hz, 2H), 3.44 (m, 3H), 3.30-2.53 (m, 8H), 2.14 (m, 7H), 1.48 (s, 3H), 1.25 (d, J = 5.5 Hz, 1H), 1.00 (m, 1H)	621.30
K12	11.09 (s, 1H), 8.11 (s, 1H), 7.59 (m, 1H), 7.15 (d, J = 8.6 Hz, 1H), 7.05 (d, J = 7.0 Hz, 1H), 6.02 (d, J = 3.4 Hz, 1H), 5.95 (t, J = 8.4 Hz, 1H), 5.57 (d, J = 3.1 Hz, 1H), 5.06 (m, 1H), 4.08 (t, J = 9.4 Hz, 1H), 3.60 (t, J = 5.4 Hz, 2H), 3.54-3.38 (m, 9H), 3.22-2.54 (m, 7H), 2.37-2.00 (m, 8H), 1.47 (s, 3H), 1.02 (t, J = 11.8 Hz, 1H)	664.51
K13	11.09 (s, 1H), 8.12 (t, J = 5.6 Hz, 1H), 7.59 (m, 1H), 7.15 (d, J = 8.6 Hz, 1H), 7.05 (d, J = 7.0 Hz, 1H), 6.02 (d, J = 3.4 Hz, 1H), 5.95 (t, J = 8.0 Hz, 1H), 5.76 (s, 1H), 5.57 (d, J = 3.1 Hz, 1H), 5.06 (m, 1H), 4.08 (t, J = 9.4 Hz, 1H), 3.65-3.36 (m, 15H), 2.77-2.57 (m, 3H), 2.37-1.52 (m, 10H), 1.47 (s, 3H), 1.28-1.21 (m, 2H)	709.40
K14	11.09 (s, 1H), 8.13 (t, J = 5.5 Hz, 1H), 7.59 (m, 1H), 7.15 (d, J = 8.6 Hz, 1H), 7.05 (d, J = 7.0 Hz, 1H), 6.04-5.89 (m, 2H), 5.57 (d, J = 3.1 Hz, 1H), 5.08-5.00 (m, 1H), 4.08 (t, J = 9.4 Hz, 1H), 3.63 (t, J = 5.4 Hz, 2H), 3.57-3.46 (m, 10H), 3.44 (s, 3H), 3.23-2.56 (m, 8H), 2.37-1.99 (m, 10H), 1.47 (s, 3H), 1.25 (d, J = 9.5 Hz, 2H)	752.49
K15	11.07 (s, 1H), 7.57 (m, 1H), 7.13 (t, J = 8.3 Hz, 2H), 6.03 (d, J = 3.5 Hz, 1H), 5.67-5.56 (m, 2H), 5.06 (m, 1H), 4.14 (t, J = 9.4 Hz, 1H), 3.55 (m, 6H), 2.88 (m, 3H), 2.74 (s, 1H), 2.34 (m, 3H), 2.22-1.52 (m, 11H), 1.50 (s, 3H), 1.42-1.01 (m, 4H)	656.56
K16	11.10 (s, 1H), 7.59 (m, 1H), 7.04 (m, 2H), 6.49 (d, J = 6.3 Hz, 1H), 6.02 (d, J = 3.5 Hz, 1H), 5.59 (m, 2H), 5.06 (m, 1H), 4.14 (t, J = 9.4 Hz, 2H), 2.96-2.82 (m, 2H), 2.69-2.55 (m, 2H), 2.35 (m, 6H), 2.23-1.96 (m, 5H), 1.76 (t, J = 9.3 Hz, 3H), 1.63 (m, 4H), 1.50 (s, 2H), 1.39 (d, J = 7.4 Hz, 1H), 1.27-1.23 (m, 3H)	656.73
K17	11.08 (s, 1H), 7.68 (m, 1H), 7.33 (m, 2H), 6.03 (d, J = 2.4 Hz, 1H), 5.65-5.51 (m, 2H), 5.09 (m, 1H), 4.14 (t, J = 9.3 Hz, 1H), 3.74 (d, J = 11.6 Hz, 2H), 2.87-2.81 (m, 3H), 2.66-2.54 (m, 3H), 2.44-1.72 (m, 13H), 1.51 (s, 3H), 1.44-1.36 (m, 3H), 1.24 (s, 3H), 1.13-1.00 (m, 3H)	684.59
K18	11.10 (s, 1H), 8.00 (t, J = 5.6 Hz, 1H), 7.81 (m, 1H), 7.48 (m, 2H), 6.02 (d, J = 3.4 Hz, 1H), 5.92 (t, J = 8.0 Hz, 1H), 5.54 (d, J = 2.9 Hz, 1H), 5.09 (m, 1H), 4.20 (t, J = 6.4 Hz, 2H), 4.10 (t, J = 9.4 Hz, 1H), 3.22 (m, 1H), 3.16 (d, J = 9.6 Hz, 1H), 3.01-2.56 (m, 5H), 2.55-2.52 (m, 1H), 2.34-2.00 (m, 7H), 1.79-1.72 (m, 2H), 1.58 (t, J = 11.3 Hz, 3H), 1.45-1.26 (m, 9H), 1.02 (m, 2H)	662.40
K19	11.06 (s, 1H), 8.00 (t, J = 5.7 Hz, 1H), 7.56 (d, J = 8.4 Hz, 1H), 7.08 (t, J = 5.3 Hz, 1H), 6.95 (d, J = 1.8 Hz, 1H), 6.85 (m, 1H), 6.03 (d, J = 3.5 Hz, 1H), 5.92 (t, J = 7.9 Hz, 1H), 5.54 (d, J = 3.1 Hz, 1H), 5.03 (m, 1H), 4.10 (t, J = 9.4 Hz, 1H), 3.26-3.12 (m, 4H), 3.02-2.52 (m, 6H), 2.37-1.51 (m, 9H), 1.48 (s, 3H), 1.34 (m, 6H), 1.24 (s, 3H)	647.50
K20	11.06 (s, 1H), 7.99 (t, J = 5.7 Hz, 1H), 7.53 (m, 1H), 7.09 (t, J = 5.2 Hz, 1H), 6.95 (d, J = 1.7 Hz, 1H), 6.85 (m, 1H), 6.02 (d, J = 3.5 Hz, 1H), 5.92 (t, J = 8.0 Hz, 1H), 5.54 (d, J = 3.1 Hz, 1H), 5.03 (m, 1H), 4.10 (t, J = 9.4 Hz, 1H), 3.25-2.52 (m, 14H), 2.37-1.96 (m, 8H), 1.64-1.53 (m, 3H), 1.24 (s, 3H), 1.21-1.16 (m, 3H), 1.02 (m, 2H)	661.80


K1		n = 3	1	6.45	10.60
K2		n = 4	1	8.85	14.41
K3		n = 5	1	19.09	19.25
K4		n = 7	1	18.83	34.13
K5		n = 3	1	N.D^b.	N.D.
K6		n = 5	1	N.D.	N.D.
K7		n = 3	1	N.D.	N.D.
K8		n = 5	1	26.16	30.63
K9		n = 7	1	2.81	4.67
K10		n = 12	1	2.03	2.50
K11		n = 1	1	38.29	N.D.
K12		n = 2	1	11.67	N.D.
K13		n = 3	1	41.67	N.D.
K14		n = 4	1	N.D.	N.D.
K15			1	4.57	9.89
K16			1	4.29	6.63
K17			1	2.11	4.60
K18		n = 8	1	3.63	5.85
K19		n = 7	2	33.81	38.06
K20		n = 8	2	5.18	8.01
FT671	--		--	8.32	N.D.


K1		n = 3	1	6.45	10.60
K2		n = 4	1	8.85	14.41
K3		n = 5	1	19.09	19.25
K4		n = 7	1	18.83	34.13
K5		n = 3	1	N.D^b.	N.D.
K6		n = 5	1	N.D.	N.D.
K7		n = 3	1	N.D.	N.D.
K8		n = 5	1	26.16	30.63
K9		n = 7	1	2.81	4.67
K10		n = 12	1	2.03	2.50
K11		n = 1	1	38.29	N.D.
K12		n = 2	1	11.67	N.D.
K13		n = 3	1	41.67	N.D.
K14		n = 4	1	N.D.	N.D.
K15			1	4.57	9.89
K16			1	4.29	6.63
K17			1	2.11	4.60
K18		n = 8	1	3.63	5.85
K19		n = 7	2	33.81	38.06
K20		n = 8	2	5.18	8.01
FT671	--		--	8.32	N.D.

小白菊内酯降解剂的设计、合成及生物学研究

PDF下载

高桐 ¹ , 张文涛 ² , 宋姗姗 ³ , 周棣 ¹ , 刘同超 ³^,^* , 缪泽鸿 ³ , 熊兵 ³^,^*

药学学报 | 研究论文 2023,58(9): 2715-2726

收起

药学学报 |研究论文 2023 , 58 (9) : 2715 -2726

小白菊内酯降解剂的设计、合成及生物学研究

全屏

[Author(id=1198960228596286264, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, orderNo=0, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=null, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1198960228709532488, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, authorId=1198960228596286264, language=EN, stringName=Tong GAO, firstName=Tong, middleName=null, lastName=GAO, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=¹, address=1. Anhui University of Chinese Medicine, Hefei 230031, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1198960228847944533, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, authorId=1198960228596286264, language=CN, stringName=高桐, firstName=桐, middleName=null, lastName=高, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=¹, address=1.安徽中医药大学, 安徽合肥 230031, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1198960227870671614, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, xref=null, ext=[AuthorCompanyExt(id=1198960227887448833, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, companyId=1198960227870671614, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1. Anhui University of Chinese Medicine, Hefei 230031, China), AuthorCompanyExt(id=1198960227900031747, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, companyId=1198960227870671614, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.安徽中医药大学, 安徽合肥 230031)])]), Author(id=1198960228982162276, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, orderNo=1, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=null, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1198960229133157237, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, authorId=1198960228982162276, language=EN, stringName=Wen-tao ZHANG, firstName=Wen-tao, middleName=null, lastName=ZHANG, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=², address=2. Shenyang Pharmaceutical University, Shenyang 110016, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1198960229254792060, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, authorId=1198960228982162276, language=CN, stringName=张文涛, firstName=文涛, middleName=null, lastName=张, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=², address=2.沈阳药科大学, 辽宁沈阳 110016, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1198960228113941264, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, xref=null, ext=[AuthorCompanyExt(id=1198960228134912786, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, companyId=1198960228113941264, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2. Shenyang Pharmaceutical University, Shenyang 110016, China), AuthorCompanyExt(id=1198960228139107091, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, companyId=1198960228113941264, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.沈阳药科大学, 辽宁沈阳 110016)])]), Author(id=1198960229355455363, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, orderNo=2, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=null, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1198960229468701586, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, authorId=1198960229355455363, language=EN, stringName=Shan-shan SONG, firstName=Shan-shan, middleName=null, lastName=SONG, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=³, address=3. Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1198960229611307934, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, authorId=1198960229355455363, language=CN, stringName=宋姗姗, firstName=姗姗, middleName=null, lastName=宋, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=³, address=3.中国科学院上海药物研究所, 上海 201203, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1198960228382376743, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, xref=null, ext=[AuthorCompanyExt(id=1198960228394959658, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, companyId=1198960228382376743, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3. Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China), AuthorCompanyExt(id=1198960228403348266, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, companyId=1198960228382376743, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3.中国科学院上海药物研究所, 上海 201203)])]), Author(id=1198960229720359851, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, orderNo=3, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=null, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1198960229821023157, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, authorId=1198960229720359851, language=EN, stringName=Di ZHOU, firstName=Di, middleName=null, lastName=ZHOU, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=¹, address=1. Anhui University of Chinese Medicine, Hefei 230031, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1198960229938463683, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, authorId=1198960229720359851, language=CN, stringName=周棣, firstName=棣, middleName=null, lastName=周, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=¹, address=1.安徽中医药大学, 安徽合肥 230031, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1198960227870671614, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, xref=null, ext=[AuthorCompanyExt(id=1198960227887448833, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, companyId=1198960227870671614, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1. Anhui University of Chinese Medicine, Hefei 230031, China), AuthorCompanyExt(id=1198960227900031747, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, companyId=1198960227870671614, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.安徽中医药大学, 安徽合肥 230031)])]), Author(id=1198960230051709903, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, orderNo=4, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=tongchao_liu@simm.ac.cn, emailSecond=null, emailThird=null, correspondingAuthor=1, authorType=1, ext={EN=AuthorExt(id=1198960230181733343, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, authorId=1198960230051709903, language=EN, stringName=Tong-chao LIU, firstName=Tong-chao, middleName=null, lastName=LIU, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=³^,^*, address=3. Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1198960230307562478, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, authorId=1198960230051709903, language=CN, stringName=刘同超, firstName=同超, middleName=null, lastName=刘, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=³^,^*, address=3.中国科学院上海药物研究所, 上海 201203, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1198960228382376743, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, xref=null, ext=[AuthorCompanyExt(id=1198960228394959658, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, companyId=1198960228382376743, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3. Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China), AuthorCompanyExt(id=1198960228403348266, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, companyId=1198960228382376743, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3.中国科学院上海药物研究所, 上海 201203)])]), Author(id=1198960230500500473, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, orderNo=5, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=null, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1198960230643105802, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, authorId=1198960230500500473, language=EN, stringName=Ze-hong MIAO, firstName=Ze-hong, middleName=null, lastName=MIAO, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=³, address=3. Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1198960230785712150, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, authorId=1198960230500500473, language=CN, stringName=缪泽鸿, firstName=泽鸿, middleName=null, lastName=缪, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=³, address=3.中国科学院上海药物研究所, 上海 201203, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1198960228382376743, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, xref=null, ext=[AuthorCompanyExt(id=1198960228394959658, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, companyId=1198960228382376743, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3. Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China), AuthorCompanyExt(id=1198960228403348266, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, companyId=1198960228382376743, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3.中国科学院上海药物研究所, 上海 201203)])]), Author(id=1198960230903152675, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, orderNo=6, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=bxiong@simm.ac.cn, emailSecond=null, emailThird=null, correspondingAuthor=1, authorType=1, ext={EN=AuthorExt(id=1198960231066730543, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, authorId=1198960230903152675, language=EN, stringName=Bing XIONG, firstName=Bing, middleName=null, lastName=XIONG, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=³^,^*, address=3. Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1198960231163199548, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, authorId=1198960230903152675, language=CN, stringName=熊兵, firstName=兵, middleName=null, lastName=熊, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=³^,^*, address=3.中国科学院上海药物研究所, 上海 201203, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1198960228382376743, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, xref=null, ext=[AuthorCompanyExt(id=1198960228394959658, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, companyId=1198960228382376743, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3. Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China), AuthorCompanyExt(id=1198960228403348266, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1198656153577161382, companyId=1198960228382376743, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3.中国科学院上海药物研究所, 上海 201203)])])]

高桐¹, 张文涛², 宋姗姗³, 周棣¹, 刘同超³^,^*, 缪泽鸿³, 熊兵³^,^*

作者信息

1.安徽中医药大学, 安徽合肥 230031

2.沈阳药科大学, 辽宁沈阳 110016

3.中国科学院上海药物研究所, 上海 201203

通讯作者:

*刘同超, Tel: 86-21-50806600-5407, E-mail: tongchao_liu@simm.ac.cn;

熊兵, Tel: 86-21-50806600-5412, E-mail: bxiong@simm.ac.cn

Synthesis, evaluation and proteomic analysis of PROTAC based on parthenolide

Tong GAO¹, Wen-tao ZHANG², Shan-shan SONG³, Di ZHOU¹, Tong-chao LIU³^,^*, Ze-hong MIAO³, Bing XIONG³^,^*

Affiliations

1. Anhui University of Chinese Medicine, Hefei 230031, China

2. Shenyang Pharmaceutical University, Shenyang 110016, China

3. Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China

出版时间: 2023-09-12 doi: 10.16438/j.0513-4870.2023-0228

文章导航

摘要

收起

小白菊内酯作为药用历史悠久的天然产物, 引起了化学家和生物学家的浓厚兴趣。现有的研究表明其具有抗炎、抗肿瘤等药理活性, 也揭示了其作用于NF-κB信号通路、DNMT1酶及Wnt/β-catenin信号通路等的生物调控功能, 但其确切的生物学靶点仍有待系统阐明。蛋白降解剂为天然产物的靶点发现提供了新的策略, 可通过蛋白组学的考察, 探究细胞中蛋白的全景变化, 从而分析其潜在的靶点。本研究基于这一思路, 以小白菊内酯为母体结构, 设计、合成了20个小白菊内酯降解剂, 测定了其体外抗肿瘤增殖活性, 并优选化合物开展蛋白组学实验, 鉴定出139个下调的差异表达蛋白(DEPs), 对小白菊内酯的作用靶点发现进行了初步探索。

关键词

小白菊内酯 / PROTAC / 靶点 / 抗肿瘤 / 蛋白组学

Abstract

收起

As a natural product with a long history of medicinal use, parthenolide has aroused great interest of chemists and biologists. Existing studies have shown that it has anti-inflammatory, antitumor and other pharmacological activities, and also revealed its action on NF-κB signaling pathway, DNMT1 enzyme and Wnt/β-catenin signaling pathway. But its biological targets remain to be elucidated systematically. Proteolysis Targeting Chimeras (PROTAC) provides a new strategy for target discovery of natural products, which can be used to explore the panorama of protein changes in cells through proteomic investigation, so as to analyze their potential targets. Based on this idea, current study designed and synthesized 20 parthenolide-derived degraders. After measured their antitumor activity in vitro, selected compounds were carried out the proteomic experiment. Finally, 139 down-regulated differentially expressed proteins were identified and the discovery of parthenolide interacting protein was preliminarily explored.

Key words

parthenolide / PROTAC / target / antitumor / proteomics

引用本文

高桐, 张文涛, 宋姗姗, 周棣, 刘同超, 缪泽鸿, 熊兵. 小白菊内酯降解剂的设计、合成及生物学研究. 药学学报, 2023 , 58 (9) : 2715 -2726 . DOI: 10.16438/j.0513-4870.2023-0228

Tong GAO, Wen-tao ZHANG, Shan-shan SONG, Di ZHOU, Tong-chao LIU, Ze-hong MIAO, Bing XIONG. Synthesis, evaluation and proteomic analysis of PROTAC based on parthenolide[J]. Acta Pharmaceutica Sinica, 2023 , 58 (9) : 2715 -2726 . DOI: 10.16438/j.0513-4870.2023-0228

正文

收起

小白菊内酯(parthenolide, PTL) 是一种倍半萜烯内酯类天然产物, 常提取于菊科(雏菊) 和木兰科(木兰属) 等药用植物中。大量研究阐述了其在抗炎、抗肿瘤、抗动脉粥样硬化等方面的生物活性^[1]。小白菊内酯及其衍生物在肺癌^[2]、结直肠癌^[3]、肝癌^[4]、骨肉瘤^[5]等多种癌症中也表现出一定的治疗潜力, 其中候选药物DMAPT已开展临床试验, 探索其替代常规化疗的治疗价值^[6]。

但同很多天然产物类似, PTL具有广泛的药理活性, 对NF-κB信号通路、DNA甲基化及细胞内氧化还原状态等生化过程均起到一定程度的调节作用^{[7, 8]}。由于其含有α-亚甲基-γ-内酯环骨架和环氧结构(图 1), PTL可以与蛋白的亲核位点形成共价结合, 故其潜在的靶点复杂多样。Michael课题组^[9]发现PTL通过泛素-蛋白酶体降解机制特异性地消耗HDAC1蛋白, 从而特异性靶向癌症干细胞; Li课题组^[10]报道了PTL抑制细胞增殖和抑制KSHV感染细胞中的NF-κB信号通路来降低KSHV致病性; Liu等^[11]在2022年首次发现了PTL能够浓度依赖性地降低了HepG2细胞中NPC1L1的表达, 这表明PTL作为一种潜在的NPC1L1抑制剂, 具有抗高胆固醇血症的治疗潜力。尽管现有研究揭示了PTL在调控生化过程中的一些作用机制, 但系统探索其确切的作用靶点仍鲜见报道。

蛋白降解靶向联合体(proteolysis targeting chimeras, PROTAC) 是一类新型药物模态(modality), 可通过生物体内的泛素-蛋白酶体系统来降解目标蛋白。故其化学结构通常由3部分组成: 目标蛋白配体分子、泛素系统中E3连接酶配体及将两者相连的连接子^[12]。与传统的小分子药物不同, 它不需要高浓度的药物剂量来占据靶点的结合口袋, 而是遵循“驱动模式”, 只需循环利用微量药物将E3连接酶与目标蛋白形成三元复合物, 进而泛素化标记目标蛋白, 通过蛋白酶体进行降解^[13-15]。随着一系列PROTAC分子(ARV-110、ARV-471、KT-474等) 进入Ⅰ/Ⅱ期临床试验, 进一步表明该类药物具有巨大的应用价值。近年来针对天然产物进行相应PROTAC分子探针的设计合成及生物活性研究也逐渐出现了^[16]。Cao课题组^[17]基于天然产物靛红(indirubin) 开发了一系列indirubin-PROTACs, 并在小鼠体内初步证实了其对HDAC6的靶向降解作用。Huang课题组^[18]设计并合成了以索拉菲尼(sorafenib) 为蛋白抑制剂的PROTACs, 通过生物学实验证实了PDEδ与索拉非尼的直接结合作用; Chen课题组^[19]报道了以乙烯二萜ZCY-001为核心支架结构, 合成ZCY-PROTACs, 发现并鉴定了MAFF是乙烯二萜的靶点。

本文针对小白菊内酯, 开展了其降解剂的设计合成, 期望通过靶向蛋白降解策略, 并结合蛋白组学初步探究其细胞中的靶点全景, 以期为天然产物靶点发现提供一个新颖的思路。

结果与讨论

收起

1 化合物的设计

由于萜类化合物特殊的骨架以及PTL不饱和内酯, 使其具有较大的改造空间。迄今, 在对PTL各个位点处进行修饰的衍生物均已得到不同程度的报道, 其中当属在C13和C14这两个位点处的改造工作居多(图 2)。Crooks课题组^[20]早在2006年, 通过Michael加成的方式在C13位点处引入二甲氨基, 大大改善了PTL的水溶性。随后Crooks和Janganati课题组^{[21, 22]}又引入了不同的胺类基团, 进一步完善在C13位进行修饰的小白菊内酯衍生物的机制研究。Zhang课题组^[23]将C14位点处的甲基通过氧化反应得到PTL羧酸衍生物; 随后Ding等^[24]首次引入了叠氮基团, 通过Click反应合成了三唑类衍生物, 并对60种肿瘤细胞系进行了筛选, 表现出了很好的抗增殖活性。基于以上研究, 设想以PTL作为目标蛋白配体设计PROTAC分子, 选择泊马度胺作为E3连接酶配体, 通过不同类型的Linker将其与PTL的C13位点及C14位点处的衍生物连接起来, 合成一系列PTL-PROTACs (图 2)。

2 化合物的合成

PTL羧酸衍生物参考已有报道的路线进行合成^[22]。在合成路线1中, 以小白菊内酯为原料, 用二氧化硒和过氧化叔丁醇进行氧化反应, 得到中间体2; 再通过Dess-Martin试剂进一步氧化, 得到醛基中间体3; 加入亚氯酸钠和2-甲基-2-丁烯进一步氧化, 得到羧酸中间体4。

目标化合物K1~K4的合成如路线2所示。商业可得原料5与不同长度的溴代酰氯通过酰化得到中间体6A~6D; 中间体6A~6D与Boc保护的哌嗪进行取代反应得到中间体7A~7D; 在酸性条件脱保护基得到中间体8A~8D; 最后在碱性环境下与PTL进行迈克尔加成反应, 得到化合物K1~K4。

中间体8A和8C分别与中间体4在碱性环境下进行酰胺缩合得到化合物K5~K6。合成步骤见路线3。

选取化合物9为起始原料, 通过与Boc保护的PEG、烷基链、饱和杂环类片段进行取代反应得到中间体10A~10K; 在酸性条件下脱去保护基得到中间体11A~11K; 最后与小白菊内酯的羧酸衍生物进行酰胺缩合, 得到化合物K7~K17。合成路线4如下。

以化合物12为起始原料, 以化合物K7~K17类似的反应路线得到化合物K18。合成步骤见路线5。

化合物K19和K20的合成如路线6所示, 以化合物15为起始原料, 反应条件参考化合物K7~K17的合成。

化合物结构经MS和¹H NMR确认, 数据见表 1。

3 活性评价

3.1 体外抗增殖活性

Li课题组^[25]在2019年揭示了PTL能够抑制USP7的活性, 并在人结肠癌细胞株HCT-116和SW480上展现了促进细胞凋亡的作用。本实验选择人结肠癌HCT-116和HT29细胞, 将文献报道的USP7抑制剂FT671^[25]作为阳性对照化合物, 采用SRB法对20个目标化合物进行HCT-116和HT29细胞增殖活性评价。表 2数据显示, 20个目标化合物对所测试的细胞均有不同程度的增殖抑制作用。其中化合物K1、K9、K10、K15、K16、K17、K18、K20对HCT-116细胞均表现出强于FT671的抗增殖活性, IC₅₀值为2.03~6.45 μmol·L^-1。

在已报道的PROTAC连接链构效关系的考察中, PEG基团常作为提高分子亲水性的优良选择^{[26, 27]}。在本实验中, 化合物K11~K14相较于其他目标化合物并没有对所测细胞株展现出一定的抑制活性; 化合物K15、K16、K17具有饱和杂环片段作为连接链, 分子具有较强的刚性结构, 表现出较好的细胞增殖抑制活性。同时, 采用烷基直链作为连接链的化合物K9和K10对HCT-116和HT29均表现出较好的抑制活性, 其中化合物K10对HCT-116和HT29的IC₅₀值分别为2.03和2.50 μmol·L^-1。

从细胞增殖活性实验的测试结果中选出抗细胞增殖能力较强的化合物K10, 进行蛋白组学考察。处理组从定量蛋白中鉴定出165个上调的差异表达蛋白(DEPs) 和139个下调的DEPs。筛选样本组内重复实验数据中至少有一半为非空值的数据, 对药物处理组(3次测定) 峰强度(intensity) 和对照组(3次测定) 峰强度进行t检验, 符合表达差异倍数大于1.5倍(上下调)且t检验小于0.05筛选标准的蛋白质视为差异表达蛋白质(图 3)。

用BIOGRID结果进行文本搜索, 发现下调蛋白与所有差异蛋白之间的PPI (protein-protein interaction networks) (图 4)。

将下调倍数排名前九的蛋白绘制散点图(图 5)。

其中CYP24A1是细胞色素P450酶家族的氧化酶之一, 负责催化1,25-双羟基维生素D3 (vitamin D3, 1,25-VD3, 1,25-dihydroxyvitamin D3, [1α,25-(OH)2D3]) 的羟基化, 将其转化为活性较低的中间产物, 并最终经一系列代谢步骤传导后排出体外, 参与了细胞增殖、分化、凋亡以及DNA修复等多个生物学环节^[28]。CYP24A1作为一种潜在的致癌基因, 它的过度表达可以激活多个信号级联反应^[29]。已有越来越多的文献报道了CYP24A1与NF-κB、VDR等信号通路之间的相互作用与关联^[30]。Li课题组^[28]发现CYP24A1可能通过NF-κB信号通路实现对Wnt/β-catenin信号通路的调控; Zhu等^[31]研究人员发现PTL可以通过结合人核糖体蛋白L10 (RPL10) 降低转录调节因子TCF4/LEF1的表达, 从而对Wnt/β-catenin信号通路产生抑制作用。本研究通过蛋白组学的考察, 发现PTL降解剂能够下调CYP24A1的表达, 不仅确证了PTL与Wnt/β-catenin通路之间的关联性, 同时也猜想, PTL可能基于下调CYP24A1的表达从而抑制Wnt/β-catenin信号通路的作用机制, 产生结肠癌细胞的抗增殖活性。

癌细胞中表观遗传格局的变化会改变基因组的结构和稳定性, 并直接导致恶性肿瘤的形成与发展。DNA甲基化是影响基因表达的主要表观遗传机制之一, 在基因表达调控、基因组印迹、X染色体失活等事件中发挥关键作用^[32]。哺乳动物DNA甲基化主要由DNA甲基转移酶(DNA methyltransferases, DNMTs) 催化甲基从S-腺苷甲硫氨酸(S-adenosylmethionine, SAM) 转移至胞嘧啶第5位碳原子^[33]。其中, DNMT1是十分关键的DNA甲基转移酶。已有文献^[34]报道了乳腺癌、胃癌常常伴随着部分肿瘤细胞中DNMT1的过表达。在Michael Goggins的研究中发现, 相较于正常细胞, 大部分胰腺癌细胞中的DNMT1数量水平显著增高^[35]。Liu等^[36]研究人员论述了PTL可以与DNA甲基转移酶活性位点共价结合并下调DNMT1蛋白的表达。本研究发现PTL降解剂显著下调了DNMT1蛋白的水平, 推测PTL可能通过抑制DNMT1从而发挥抗肿瘤作用。

双特异性酪氨酸磷酸化调节激酶(CDC-like Kinases, CLK) 是一种作用于丝氨酸/苏氨酸和酪氨酸的双特异性激酶。其中CLK3作为CDK家族的一个亚型, 通过磷酸化富含丝氨酸/精氨酸的蛋白质如SRSF1和SRSF3来调节RNA剪接, 多项研究表明这一过程的失调与癌症、神经疾病以及多种遗传疾病息息相关^[37]。Zhang课题组^[38]发现胆管癌细胞中常常伴随着CLK3的异常表达, 揭示了CLK3突变体通过激活嘌呤合成信号通路促进胆管癌细胞发育的相关机制。本研究中, 小白菊内酯降解剂下调CLK3蛋白的表达水平也说明小白菊内酯可能通过抑制CLK3发挥抗结肠癌细胞功能。

用DAVID网站对下调蛋白进行在线富集分析, 获得每个蛋白相关的功能词条, 发现这些蛋白的功能比较集中于线粒体和核糖体功能, 以及RNA结合区域。结合Wang课题组^[39]对PTL代谢组学的考察, 推测PTL可能通过能量代谢的方式参与对肿瘤细胞的调控。此外, Li课题组^{[40, 41]}相继报道了PTL对USP7和USP47的抑制作用。USP10是泛素－蛋白酶体系统(ubiquitin-proteasome system, UPS) 中的去泛素化酶之一, 发现PTL降解剂同样对其产生了一定程度上的抑制。基于此, 推测PTL抑制结肠癌细胞活性的同时, 也能够借助UPS系统在一定程度上促进肿瘤细胞的凋亡。

小结

收起

基于小白菊内酯的化学结构及已有报道针对其衍生物所进行的合成工作的报道, 本实验设计并合成了20个小白菊内酯降解剂。采用SRB法评价化合物对人结肠癌HCT-116和HT29细胞的增殖抑制作用。结果表明, 大多数化合物具有一定的抗肿瘤增殖活性, 其中化合物K10对HCT-116细胞的IC₅₀值为2.03 μmol·L^-1。在细胞水平上, 选用Exploris 480对K10进行蛋白组学分析, 从定量蛋白中鉴定出165个上调的DEPs和139个下调的DEPs, 发现了DNMT1、CLK3、MRPs等鲜有报道的相关蛋白, 为小白菊内酯作用靶点的发现提供了新颖的思路。

实验部分

收起

柱色谱硅胶(200~300目) 为青岛海洋化工厂生产, 薄层色谱板采用254 nm紫外灯检测。¹H NMR和¹³C NMR采用Bruker 400和Bruker 101核磁共振仪测定。所有试剂均为市售分析纯或化学纯, 除特别说明外, 一般不经纯化处理直接使用。

1 化学合成

1.1 中间体2的合成

取单口瓶, 依次加入小白菊内酯(1 g, 4.02 mmol)、干燥二氯甲烷25 mL、二氧化硒(716 mg, 6.40 mmol), 在氮气保护下加入预干燥的过氧化叔丁醇(1.50 mL, 9.70 mmol), 继续反应18 h, TLC监测反应完毕, 加入二氯甲烷将反应液稀释至80 mL, 饱和硫代硫酸钠水溶液20 mL洗3次, 无水硫酸钠干燥, 抽滤, 减压蒸干溶剂, 得到粗品。硅胶色谱[V_石油醚∶V_乙酸乙酯 = 1∶1] 纯化得到白色固体即为中间体2, 产率为60%。

1.2 中间体3的合成

取单口瓶依次按顺序加入中间2 (100 mg, 0.37 mmol)、干燥二氯甲烷15 mL、碳酸氢钠(320 mg, 3.8 mmol), 在氮气保护下分批加入Dess-Martin试剂(240 mg, 0.57 mmol)。室温搅拌2 h反应完毕。加入二氯甲烷将反应液稀释至80 mL, 饱和硫代硫酸钠水溶液20 mL洗3次, 无水硫酸钠干燥, 抽滤, 减压蒸干溶剂, 得到粗品。硅胶色谱[V_石油醚∶V_乙酸乙酯 = 2∶1] 纯化得到白色固体即为中间体3, 产率70%。

1.3 中间体4的合成

取单口瓶依次按顺序加入中间体3 (200 mg, 0.76 mmol)、叔丁醇10 mL、纯水3.5 mL、2-甲基-2-丁烯220 μL、亚氯酸钠(140 mg, 1.54 mmol)、磷酸二氢钠(900 mg, 7.50 mmol), 在氮气保护下过夜搅拌。反应结束后向反应液中加入纯水40 mL, 加入乙酸乙酯80 mL, 萃取3次。无水硫酸钠干燥, 抽滤, 减压蒸干溶剂, 得到粗品。硅胶色谱[V_石油醚∶V_乙酸乙酯 = 1∶1~2] 得到白色固体即为中间体4, 产率70%。

1.4 中间体6A~6D的合成

向三口烧瓶中加入化合物5 (250 mg, 0.92 mmol)、干燥四氢呋喃5 mL, 氮气保护下缓慢加入相应的溴代酰氯(3.66 mmol), 加毕升温至70 ℃回流, 反应1 h, TLC监测反应进程。反应结束后体系自然回温至室温, 加入甲醇搅拌30 min, 搅拌过程中逐渐有固体析出。将固液混合物转移至单口瓶减压蒸干溶剂得到固体, 加入乙酸乙酯5 mL, 石油醚1 mL打浆, 抽滤, 得到中间体6A~6D, 均为淡黄色固体, 产率60%。

1.5 中间体7A~7D的合成

取三口瓶, 依次按顺序加入中间体6A/6B/6C/6D (300 mg)、1-N-Boc-哌嗪(3.39 mmol)、无水乙腈8 mL、三乙胺(1.02 mmol), 升温至60 ℃回流, 3 h反应完毕。向反应体系加入乙酸乙酯40 mL, 饱和氯化铵水溶液5 mL, 洗2次, 有机相无水硫酸钠干燥, 抽滤。减压蒸干溶剂, 硅胶色谱[V_二氯甲烷∶V_甲醇 = 100∶1] 分离得到淡黄色固体, 产率50%。

1.6 中间体8A~8D的合成

取单口瓶, 依次加入中间体7A/7B/7C/7D (80 mg)、无水二氯甲烷2 mL、盐酸二氧六环1.5 mL, 室温反应1 h, 反应完毕。减压蒸干溶剂, 得到淡黄色固体, 产率90%。

1.7 化合物K1~K4的合成

取单口瓶, 依次加入小白菊内酯(25 mg, 0.10 mmol)、中间体8A~8D (0.10 mmol)、无水乙醇4 mL、三乙胺(15 μL, 0.10 mmol), 氮气保护下搅拌过夜。加入二氯甲烷将反应液稀释至60 mL, 饱和氯化铵水溶液15 mL洗3次。无水硫酸钠干燥, 抽滤, 减压蒸干溶剂得到粗品。柱色谱纯化[V_二氯甲烷∶V_甲醇 = 50∶1~40∶1] 得到化合物K1~K4, 均为白色固体, 产率60%。

1.8 化合物K5、K6的合成

取单口瓶, 依次加入中间体4 (30 mg, 0.11 mmol)、无水二氯甲烷3 mL、N, N-二异丙基乙胺(40 μL, 0.22 mmol), 室温搅拌20 min, 加入中间体8A~8D (50 mg, 0.11 mmol)、2-(7-氮杂苯并三氮唑)-N, N, N′, N′-四甲基脲六氟磷酸酯(82 mg, 0.22 mmol), 氮气保护下过夜搅拌。反应完毕, 向反应液中加入二氯甲烷80 mL, 饱和氯化铵水溶液20 mL洗3次, 无水硫酸钠干燥, 抽滤, 减压蒸干溶剂, 得到粗品。柱色谱纯化[V_二氯甲烷∶V_甲醇 = 50∶1~40∶1] 得到化合物K5和K6, 均为白色固体, 产率60%。

1.9 中间体10A~10K的合成

取单口瓶依次加入中间体10 (250 mg, 0.91 mmol), 干燥N, N-二甲基甲酰胺5 mL, Boc保护的PEG、烷基链、饱和杂环结构(0.91 mmol), N, N-二异丙基乙胺升温至80 ℃回流。搅拌12 h, 反应结束。反应装置回温至室温, 加入乙酸乙酯80 mL, 饱和氯化锂水溶液洗(6~8次), 直至将反应体系中的N, N-二甲基甲酰胺除净。无水硫酸钠干燥有机相, 抽滤, 减压蒸干溶剂, 得到粗品。柱色谱纯化[V_二氯甲烷∶V_甲醇 = 90∶1~80∶1] 得到中间体10A~10K, 均为黄绿色固体, 产率60%。

1.10 中间体11A~11K的合成

取单口瓶, 依次加入中间体10A~10K (100 mg)、无水二氯甲烷2 mL、盐酸二氧六环1.5 mL, 室温反应1 h, 反应完毕。减压蒸干溶剂, 得到中间体11A~11K, 均为淡黄色固体, 产率90%。

1.11 化合物K7~K17的合成

采用与化合物K5和K6类似的方法, 得到化合物K7~K17, 均为淡黄色固体, 产率60%。

1.12 中间体13的合成

取单口瓶, 依次加入中间体12 (225 mg, 0.76 mmol)、N, N-二甲基甲酰胺3 mL、碘化钾(13 mg, 0.08 mmol)、碳酸氢钾(114 mg, 1.14 mmol)、1-叔丁氧羰基-1,8-二氨基辛烷(250 mg, 0.96 mmol)。升温至60 ℃, 回流4 h, 反应结束。反应装置回温至室温, 反应液中加入乙酸乙酯80 mL, 饱和氯化锂水溶液12 mL洗(6~8次), 直至将反应体系中的N, N-二甲基甲酰胺除净。无水硫酸钠干燥有机相, 抽滤, 减压蒸干溶剂, 得到粗品。柱色谱纯化[V_二氯甲烷∶V_甲醇 = 90∶1~80∶1] 得到黄绿色固体, 产率60%。

1.13 中间体14的合成

取单口瓶, 依次加入中间体13 (100 mg, 0.19 mmol)、无水二氯甲烷2 mL、盐酸二氧六环1.5 mL, 室温反应1 h, 反应完毕。减压蒸干溶剂, 得到淡黄色固体, 产率90%。

1.14 化合物K18的合成

取单口瓶, 依次加入中间体4 (30 mg, 0.11 mmol)、无水二氯甲烷3 mL、N, N-二异丙基乙胺(40 μL, 0.22 mmol), 室温搅拌20 min, 加入中间体14 (45 mg, 0.11 mmol)、2-(7-氮杂苯并三氮唑)-N, N, N′, N′-四甲基脲六氟磷酸酯(82 mg, 0.22 mmol), 氮气保护下过夜搅拌。反应完毕, 向反应液中加入二氯甲烷80 mL, 饱和氯化铵水溶液20 mL洗3次, 无水硫酸钠干燥, 抽滤, 减压蒸干溶剂, 得到粗品。柱色谱纯化[V_二氯甲烷∶V_甲醇 = 50∶1~40∶1] 得到淡黄色固体, 产率60%。

1.15 化合物K19、K20的合成

与化合物K7~K17类似的方法, 得到淡黄色固体K19和K20, 产率60%。

2 生物部分

2.1 体外抗增殖作用评价

McCoy's 5A培养基和胎牛血清购自美国Gibco公司; 磺酰罗单明B (sulforodamine B, SRB) 购自Sigma-Aldrich公司; 三氯醋酸(TCA)、冰醋酸(HAC) 及Tris均购自国药集团。人结肠癌细胞HCT-116及HT29均购自American type culture collection (ATCC) 并按要求培养。

采用SRB法进行体外抗增殖活性研究, 具体步骤如下: 将处于对数生长期的HCT-116和HT29细胞以90 μL (每孔1 200个和1 500个) 接种于96孔培养板, 37 ℃培养过夜。每孔加入10 μL一定稀释浓度的药物, 每个浓度设3个重复, 并设相应浓度的生理盐水溶媒对照及无细胞调零孔。加药后肿瘤细胞在37 ℃、5% CO₂条件下培养3天。弃去培养液, 加入4 ℃预冷的10%的三氯醋酸(TCA) (每孔100 μL), 4 ℃固定1 h后用蒸馏水洗涤5次, 65 ℃烘箱热风干燥1 h。再加入由1%冰醋酸配制的SRB (4 mg·mL^-1) 溶液(每孔100 μL), 室温中染色15 min。而后去上清液, 用1%醋酸洗涤5次, 65 ℃烘箱热风干燥1 h。每孔加入150 μL的Tris (10 mmol·L^-1) 溶液, 室温放置15 min。SPECTAR MAX190酶标仪560 nm波长下测定OD值。按下列公式计算被测物对癌细胞生长的抑制率, 半数抑制量IC₅₀值采用Logit法计算。抑制率= (对照组OD值－给药组OD值)/对照组OD值×100%, 计算IC₅₀。

2.2 蛋白组学细胞处理

细胞样品准备: 将处于对数生长期的HCT-116细胞以每孔4.5×10⁵个接种于6孔细胞培养板中, 37 ℃培养过夜待细胞贴壁。每孔更换1 800 μL新鲜培养基后加入200 μL K10 (终浓度为10 μmol·L^-1), 设3个重复并设置3个DMSO对照孔。加药后细胞在37 ℃、5% CO₂条件下培养24 min。弃掉培养液, 加入4 ℃预冷的生理盐水, 平放轻轻摇动1 min洗涤细胞, 倒掉生理盐水。重复清洗两次后, 将培养皿置于冰上, 继续加入4 ℃预冷的生理盐水, 然后用干净的细胞刮棒快速收集细胞, 移液管吸取至预冷的冻存管内。最后, 4 ℃、1 000 g离心5 min, 去上清, 最终收集细胞沉淀于冻存管中。

2.3 质谱分析

提取肽段在C18容器上脱盐, 然后真空离心浓缩, 在40 µL 0.1% (v/v) 甲酸中重建。根据脊椎动物蛋白质中色氨酸和酪氨酸的频率计算, 在0.1% (g·L^-1) 溶液中使用1.1的消光系数, 在280 nm处通过紫外光谱密度估算肽含量。LC-MS/MS分析在Orbitrap Exploris 480质谱仪(Thermo Fisher Scientific) 上进行, 该质谱仪与Easy nLC (Thermo Fisher Scientific) 耦合。2 μL肽在缓冲液A (0.1%甲酸) 中加载到C18反相分析柱(Thermo Fisher Scientific, Acclaim PepMap RSLC 50 μm X 150 mm, nano viper, P/N164943) 上, 以缓冲液B (80%乙腈和0.1%甲酸) 线性梯度分离, 流速为300 nL·min^-1。

使用数据依赖的top10方法获取MS数据, 从调查扫描(350~1 200 m/z) 中动态选择最丰富的前体离子用于HCD碎片。MS1扫描在m/z 200的分辨率为120 000, AGC目标为300%, 最大IT为50 ms。数据依赖模式为周期时间, 周期时间设置为1.5 s。MS2扫描在m/z 200的分辨率为15 000, AGC目标为75%, 最大IT为35 ms, 隔离宽度为1.6 m/z。Microscans设置为1。只有电荷在2~6之间的离子。所选离子的动态排除时间为30秒。归一化碰撞能量为33%。

2.4 数据分析

MS数据采用MaxQuant软件版本1.6.17.0进行分析。在数据库中搜索MS数据(由项目确定)。最初的搜索设定在6 ppm的前体质量窗口。搜索遵循胰蛋白酶/P的酶促裂解规则, 允许最大两个缺失的裂解位点和对片段离子的质量耐受为20 ppm。将半胱氨酸氨基甲基化定义为固定修饰, 将蛋白质N端乙酰化和蛋氨酸氧化定义为可变修饰, 便于数据库检索。肽和蛋白质鉴定的全球错误发现率(FDR) 的截止值设置为0.01。蛋白质丰度根据归一化光谱蛋白质强度(LFQ强度) 计算。Fold变化为2或0.5, P值为0.05的蛋白被认为是差异表达蛋白。

2.5 生物信息学

在Linux服务器上使用NCBI BLAST + (NCBI-BLAST-2.3.0+) 将所有蛋白质序列比对到数据库, 只保留前10位和E-value<=1e-3的序列。其次, 选择GO术语(数据库版本: go_20190701。由Blast2GO的顶级比特分数序列的obo)。然后通过Blast2GO Command Line完成GO术语到蛋白质的标注。在基本注释完成后, 利用InterProScan根据motif搜索EBI数据库, 然后将motif的功能信息添加到蛋白质中以提高注释, 再由ANNEX对GO术语之间的注释和连接进行了进一步的改进。采用Fisher精确检验(Fisher's Exact Test), 通过比较与GO项相关的差异表达蛋白数量和总蛋白数量来丰富GO项。

通过KEGG数据库[数据库版本: KO_INFO_END.txt (2022.11.05)] 进行通路分析。通过比较与通路相关的差异表达蛋白数量和总蛋白数量, 使用Fisher's Exact Test来识别显著富集的通路。

作者贡献: 高桐负责完成相关实验合成工作; 张文涛协助完成合成工作及化合物鉴定工作; 宋姗姗负责活性测定实验; 周棣协助完成化合物鉴定工作; 刘同超协助完成论文撰写等工作; 缪泽鸿和熊兵负责选题、实验指导和论文撰写。

利益冲突: 作者声明本文不存在任何利益冲突。

参考文献

收起

文献

收起

参考文献引证文献

排序方式：

[1]

Végh K, Alberti Á, Riethmüller E, et al. Supercritical fluid extraction and convergence chromatographic determination of parthenolide in Tanacetum parthenium L.: experimental design, modeling and optimization[J]. J Supercrit Fluids, 2014, 95: 84-91.

https://doi.org/10.1016/j.supflu.2014.07.029

[2]

Talib WH, Al Kury LT. Parthenolide inhibits tumor-promoting effects of nicotine in lung cancer by inducing P53-dependent apoptosis and inhibiting VEGF expression[J]. Biomed Pharmacother, 2018, 107: 1488-1495.

https://doi.org/10.1016/j.biopha.2018.08.139

[3]

Kim SL, Trang KT, Kim SH, et al. Parthenolide suppresses tumor growth in a xenograft model of colorectal cancer cells by inducing mitochondrial dysfunction and apoptosis[J]. Int J Oncol, 2012, 41: 1547-1553.

https://doi.org/10.3892/ijo.2012.1587

[4]

Carlisi D, D′Anneo A, Angileri L, et al. Parthenolide sensitizes hepatocellular carcinoma cells to trail by inducing the expression of death receptors through inhibition of STAT3 activation[J]. J Cell Physiol, 2011, 226: 1632-1641.

https://doi.org/10.1002/jcp.22494

[5]

Mendonca MS, Turchan WT, Alpuche ME, et al. DMAPT inhibits NF-kappa B activity and increases sensitivity of prostate cancer cells to X-rays in vitro and in tumor xenografts in vivo[J]. Free Radical Biol Med, 2017, 112: 318-326.

https://doi.org/10.1016/j.freeradbiomed.2017.08.001

[6]

Liu X, Wang X. Recent advances on the structural modification of parthenolide and its derivatives as anticancer agents[J]. Chin J Nat Med, 2022, 20: 814-829.

[7]

Freund RRA, Gobrecht P, Fischer D, et al. Advances in chemistry and bioactivity of parthenolide[J]. Nat Prod Rep, 2020, 37: 541-565.

https://doi.org/10.1039/C9NP00049F

[8]

Freund RRA, Gobrecht P, Rao Z, et al. Stereoselective total synthesis of parthenolides indicates target selectivity for tubulin carboxypeptidase activity[J]. Chem Sci, 2019, 10: 7358-7364.

https://doi.org/10.1039/C9SC01473J

[9]

Gopal YV, Arora TS, Van Dyke MW. Parthenolide specifically depletes histone deacetylase 1 protein and induces cell death through ataxia telangiectasia mutated[J]. Chem Biol, 2007, 14: 813-823.

https://doi.org/10.1016/j.chembiol.2007.06.007

[10]

Li Y, Xu H, Tan X, et al. Parthenolide inhibits proliferation of cells infected with Kaposi′s sarcoma-associated herpesvirus by suppression of the NF-κB signaling pathway[J]. Arch Virol, 2023, 168: 39.

https://doi.org/10.1007/s00705-022-05626-0

[11]

Liu W, Liang B, Zeng J, et al. First discovery of cholesterol-lowering activity of parthenolide as NPC1L1 inhibitor[J]. Molecules, 2022, 27: 6270.

https://doi.org/10.3390/molecules27196270

[12]

Li XY, Pu WC, Zheng QQ, et al. Proteolysis-targeting chimeras (PROTACs) in cancer therapy[J]. Mol Cancer, 2022, 21: 99.

https://doi.org/10.1186/s12943-021-01434-3

[13]

Bekes M, Langley DR, Crews CM. PROTAC targeted protein degraders: the past is prologue[J]. Nat Rev Drug Discov, 2022, 21: 181-200.

https://doi.org/10.1038/s41573-021-00371-6

[14]

Fisher SL, Phillips AJ. Targeted protein degradation and the enzymology of degraders[J]. Curr Opin Chem Biol, 2018, 44: 47-55.

https://doi.org/10.1016/j.cbpa.2018.05.004

[15]

Kozicka Z, Thoma NH. Haven′t got a glue: protein surface variation for the design of molecular glue degraders[J]. Cell Chem Biol, 2021, 28: 1032-1047.

https://doi.org/10.1016/j.chembiol.2021.04.009

[16]

Bian J, Ren J, Li Y, et al. Discovery of Wogonin-based PROTACs against CDK9 and capable of achieving antitumor activity[J]. Bioorg Chem, 2018, 81: 373-381.

https://doi.org/10.1016/j.bioorg.2018.08.028

[17]

Cao ZX, Gu ZC, Lin SX, et al. Attenuation of NLRP3 inflammasome activation by indirubin-derived PROTAC targeting HDAC6[J]. ACS Chem Biol, 2021, 16: 2746-2751.

https://doi.org/10.1021/acschembio.1c00681

[18]

Li Y, Meng Q, Wang P, et al Identification of PDE6D as a potential target of sorafenib via PROTAC technology bioRxiv 2020. DOI: 10.1101/2020.05.06.079947.

https://doi.org/10.1101/2020.05.06.079947

[19]

Wu YL, Yang YY, Wang W, et al. PROTAC technology as a novel tool to identify the target of lathyrane diterpenoids[J]. Acta Pharm Sin B, 2022, 12: 4262-4265.

https://doi.org/10.1016/j.apsb.2022.07.007

[20]

Nasim S, Crooks PA. Antileukemic activity of aminoparthenolide analogs[J]. Bioorg Med Chem Lett, 2008, 18: 3870-3873.

https://doi.org/10.1016/j.bmcl.2008.06.050

[21]

Janganati V, Penthala NR, Cragle CE, et al. Heterocyclic aminoparthenolide derivatives modulate G(2)-M cell cycle progression during Xenopus oocyte maturation[J]. Bioorg Med Chem Lett, 2014, 24: 1963-1967.

https://doi.org/10.1016/j.bmcl.2014.02.067

[22]

Yang ZJ, Ge WZ, Li QY, et al. Syntheses and biological evaluation of costunolide, parthenolide, and their fluorinated analogues[J]. J Med Chem, 2015, 58: 7007-7020.

https://doi.org/10.1021/acs.jmedchem.5b00915

[23]

Zeng BL, Cheng Y, Zheng KL, et al. Design, synthesis and in vivo anticancer activity of novel parthenolide and micheliolide derivatives as NF-kB and STAT3 inhibitors[J]. Bioorg Chem, 2021, 111: 104973.

https://doi.org/10.1016/j.bioorg.2021.104973

[24]

Ding YH, Li SZ, Ge Z, et al. Design and synthesis of parthenolide and 5-fluorouracil conjugates as potential anticancer agents against drug resistant hepatocellular carcinoma[J]. Eur J Med Chem, 2019, 183: 111706.

https://doi.org/10.1016/j.ejmech.2019.111706

[25]

Li X, Kong L, Yang Q, et al. Parthenolide inhibits ubiquitin-specific peptidase 7 (USP7), Wnt signaling, and colorectal cancer cell growth[J]. J Biol Chem, 2020, 295: 3576-3589.

https://doi.org/10.1074/jbc.RA119.011396

[26]

Turnbull AP, Ioannidis S, Krajewski WW, et al. Molecular basis of USP7 inhibition by selective small-molecule inhibitors[J]. Nature, 2017, 550: 481-486.

https://doi.org/10.1038/nature24451

[27]

Rana S, Bendjennat M, Kour S, et al. Selective degradation of CDK6 by a palbociclib based PROTAC[J]. Bioorg Med Chem Lett, 2019, 29: 1375-1379.

https://doi.org/10.1016/j.bmcl.2019.03.035

[28]

Chen XQ, Mao JY, Wang CS, et al. CYP24A1 involvement in inflammatory factor regulation occurs via the Wnt signaling pathway[J]. Curr Med Sci, 2022, 42: 1022-1032.

https://doi.org/10.1007/s11596-022-2564-x

[29]

Sun H, Jiang C, Cong L, et al. CYP24A1 inhibition facilitates the antiproliferative effect of 1, 25(OH)2D3 through downregulation of the WNT/β-catenin pathway and methylation-mediated regulation of CYP24A1 in colorectal cancer cells[J]. DNA Cell Biol, 2018, 37: 742-749.

https://doi.org/10.1089/dna.2017.4058

[30]

Anderson MG, Nakane M, Ruan X, et al. Expression of VDR and CYP24A1 mRNA in human tumors[J]. Cancer Chemother Pharmacol, 2006, 57: 234-240.

https://doi.org/10.1007/s00280-005-0059-7

[31]

Zhu X, Yuan C, Tian C, et al. The plant sesquiterpene lactone parthenolide inhibits Wnt/beta-catenin signaling by blocking synthesis of the transcriptional regulators TCF4/LEF1[J]. J Biol Chem, 2018, 293: 5335-5344.

https://doi.org/10.1074/jbc.M117.819300

[32]

Klutstein M, Nejman D, Greenfield R, et al. DNA methylation in cancer and aging[J]. Cancer Res, 2016, 76: 3446-3450.

[33]

Ren W, Gao L, Song J. Structural basis of DNMT1 and DNMT3A-mediated DNA methylation[J]. Genes (Basel), 2018, 9: 620-639.

https://doi.org/10.3390/genes9120620

[34]

Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation[J]. Cell, 2011, 144: 646-674.

https://doi.org/10.1016/j.cell.2011.02.013

[35]

Li A, Omura N, Hong SM, et al. Pancreatic cancer DNMT1 expression and sensitivity to DNMT1 inhibitors[J]. Cancer Biol Ther, 2010, 9: 321-329.

https://doi.org/10.4161/cbt.9.4.10750

[36]

Liu Z, Liu S, Xie Z, et al. Modulation of DNA methylation by a sesquiterpene lactone parthenolide[J]. J Pharmacol Exp Ther, 2009, 329: 505-514.

https://doi.org/10.1124/jpet.108.147934

[37]

Lee JY, Yun JS, Kim WK, et al. Structural basis for the selective inhibition of Cdc2-like kinases by CX-4945[J]. Biomed Res Int, 2019, 2019: 6125068.

[38]

Zhou Q, Lin M, Feng X, et al. Targeting CLK3 inhibits the progression of cholangiocarcinoma by reprogramming nucleotide metabolism[J]. J Exp Med, 2020, 217: e20191779.

https://doi.org/10.1084/jem.20191779

[39]

Yuan L, Wang Z, Zhang D, et al. Metabonomic study of the intervention effects of parthenolide on anti-thyroid cancer activity[J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2020, 1150: 122179.

https://doi.org/10.1016/j.jchromb.2020.122179

[40]

Li X, Kong L, Yang Q, et al. Parthenolide inhibits ubiquitin-specific peptidase 7 (USP7), Wnt signaling, and colorectal cancer cell growth[J]. J Biol Chem, 2020, 295: 3576-3589.

https://doi.org/10.1074/jbc.RA119.011396

[41]

Zhang S, Ju X, Yang Q, et al. USP47 maintains the stemness of colorectal cancer cells and is inhibited by parthenolide[J]. Biochem Biophys Res Commun, 2021, 562: 21-28.

https://doi.org/10.1016/j.bbrc.2021.05.017

2023年第58卷第9期

PDF下载

520

247

引用本文

BibTeX

文章信息

doi: 10.16438/j.0513-4870.2023-0228

接收时间：2023-02-27
首发时间：2025-11-21
出版时间：2023-09-12

补充材料

相关文章

文章信息

作者

出版历史

收稿日期：2023-02-27
修回日期：2023-03-28

基金

作者信息

1.安徽中医药大学, 安徽合肥 230031

2.沈阳药科大学, 辽宁沈阳 110016

3.中国科学院上海药物研究所, 上海 201203

通讯作者:

*刘同超, Tel: 86-21-50806600-5407, E-mail: tongchao_liu@simm.ac.cn;

熊兵, Tel: 86-21-50806600-5412, E-mail: bxiong@simm.ac.cn

参考文献

分享链接

https://castjournals.cast.org.cn/joweb/yxxb/CN/10.16438/j.0513-4870.2023-0228

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

Compd.	¹H NMR (400 MHz, DMSO-d₆)	ESI-MS (m/z) [M+H]⁺
K1	11.16 (s, 1H), 9.69 (s, 1H), 8.51 (d, J = 8.4 Hz, 1H), 7.84 (t, J = 7.9 Hz, 1H), 7.61 (d, J = 7.2 Hz, 1H), 5.26-5.12 (m, 2H), 3.94 (t, J = 9.1 Hz, 1H), 2.77 (d, J = 9.1 Hz, 1H), 2.59 (m, 4H), 2.47 (d, J = 7.0 Hz, 2H), 2.25-1.56 (m, 17H), 1.32 (d, J = 15.5 Hz, 2H), 1.25 (d, J = 10.2 Hz, 6H), 1.20 (s, 3H), 1.12 (m, 2H)	675.46
K2	11.15 (s, 1H), 9.70 (s, 1H), 8.48 (d, J = 8.4 Hz, 1H), 7.87-7.81 (m, 1H), 7.62 (d, J = 7.1 Hz, 1H), 5.22-5.07 (m, 2H), 3.96 (t, J = 9.1 Hz, 1H), 2.78 (d, J = 9.1 Hz, 1H), 2.61 (m, 5H), 2.43-1.92 (m, 17H), 1.63 (d, J = 9.3 Hz, 6H), 1.55-1.48 (m, 3H), 1.24 (s, 3H), 1.20 (s, 2H), 0.89-0.81 (m, 2H)	689.57
K3	11.15 (s, 1H), 9.70 (s, 1H), 8.48 (d, J = 8.4 Hz, 1H), 7.92-7.80 (m, 1H), 7.62 (d, J = 7.2 Hz, 1H), 5.26-5.11 (m, 2H), 3.96 (t, J = 9.1 Hz, 1H), 2.96-2.85 (m, 1H), 2.78 (d, J = 9.1 Hz, 1H), 2.60 (m, 6H), 2.48-1.91 (m, 17H), 1.74-1.56 (m, 6H), 1.48 (s, 2H), 1.33 (m, 3H), 1.20 (s, 3H), 1.11 (m, 2H)	703.64
K4	11.16 (s, 1H), 9.69 (s, 1H), 8.48 (d, J = 8.4 Hz, 1H), 7.89-7.78 (m, 1H), 7.62 (d, J = 7.0 Hz, 1H), 5.24-5.10 (m, 2H), 3.96 (t, J = 9.1 Hz, 1H), 2.79 (d, J = 9.1 Hz, 1H), 2.67-2.58 (m, 4H), 2.49-2.44 (m, 4H), 2.42-1.91 (m, 14H), 1.69-1.23 (m, 19H), 1.20 (s, 3H)	732.60
K5	11.15 (s, 1H), 9.70 (s, 1H), 8.50 (d, J = 8.4 Hz, 1H), 7.90-7.79 (m, 1H), 7.62 (d, J = 7.3 Hz, 1H), 6.03 (d, J = 3.4 Hz, 1H), 5.65-5.53 (m, 2H), 5.15 (m, 1H), 4.14 (t, J = 9.4 Hz, 1H), 3.18 (s, 1H), 2.96-2.83 (m, 2H), 2.61 (m, 2H), 2.48-1.57 (m, 19H), 1.50 (s, 2H), 1.43-1.29 (m, 1H), 1.28-1.21 (m, 2H), 1.16-0.98 (m, 2H)	687.30
K6	11.16 (s, 1H), 9.70 (s, 1H), 8.48 (t, J = 6.9 Hz, 1H), 7.84 (t, J = 7.9 Hz, 1H), 7.62 (d, J = 7.2 Hz, 1H), 6.68 (d, J = 8.3 Hz, 1H), 6.03 (d, J = 3.4 Hz, 1H), 5.57 (d, J = 3.0 Hz, 1H), 5.15 (m, 1H), 4.15 (t, J = 9.4 Hz, 1H), 2.37-1.98 (m, 14H), 1.64 (d, J = 7.9 Hz, 4H), 1.32 (d, J = 14.5 Hz, 6H), 1.25 (t, J = 6.6 Hz, 11H)	715.30
K7	11.10 (s, 1H), 8.09 (t, J = 5.7 Hz, 1H), 7.64-7.52 (m, 1H), 7.08-6.99 (m, 2H), 6.66 (t, J = 6.0 Hz, 1H), 5.98 (t, J = 5.6 Hz, 2H), 5.54 (d, J = 3.0 Hz, 1H), 4.17 (m, 3H), 3.14 (m, 2H), 2.65-2.54 (m, 2H), 2.15 (m, 7H), 1.74-1.62 (m, 4H), 1.49 (s, 3H), 1.25 (m, 4H)	591.20
K8	11.09 (s, 1H), 8.02 (s, 1H), 7.65-7.52 (m, 1H), 7.06 (m, 2H), 6.51 (t, J = 5.6 Hz, 1H), 6.02 (d, J = 3.4 Hz, 1H), 5.97-5.87 (m, 1H), 5.56 (d, J = 2.4 Hz, 1H), 5.05 (m, 1H), 4.11 (t, J = 9.4 Hz, 1H), 3.27-3.15 (m, 3H), 2.95-2.82 (m, 1H), 2.65-2.53 (m, 2H), 2.36-1.53 (m, 10H), 1.48 (s, 3H), 1.45-1.23 (m, 8H)	647.50
K9	11.09 (s, 1H), 7.59 (m, 1H), 7.06 (m, 2H), 6.02 (d, J = 3.4 Hz, 1H), 5.92 (t, J = 7.9 Hz, 1H), 5.54 (d, J = 3.1 Hz, 1H), 5.05 (m, 1H), 4.10 (t, J = 9.4 Hz, 1H), 3.30-2.56 (m, 12H), 2.35-1.52 (m, 12H), 1.48 (s, 3H), 1.24 (s, 3H), 1.03 (t, J = 12.6 Hz, 1H), 0.88-0.80 (m, 2H)	646.74
K10	11.09 (s, 1H), 8.14-7.83 (m, 1H), 7.58 (m, 2H), 7.06 (m, 3H), 6.51 (d, J = 5.3 Hz, 1H), 6.14-5.80 (m, 3H), 5.76 (s, 1H), 5.53 (d, J = 3.1 Hz, 2H), 5.03 (m, 2H), 4.05 (m, 2H), 3.31-3.25 (m, 3H), 3.24-3.11 (m, 2H), 3.03-2.91 (m, 2H), 2.91-2.81 (m, 1H), 2.80-2.65 (m, 2H), 2.65-2.54 (m, 3H), 2.32 (d, J = 12.1 Hz, 2H), 2.24 (m, 3H), 2.22-2.14 (m, 2H), 2.10 (m, 2H), 2.04 (m, 1H), 1.59 (t, J = 15.4 Hz, 5H), 1.48 (s, 4H), 0.84 (m, 2H)	716.75
K11	11.10 (s, 1H), 8.12 (s, 1H), 7.60 (m, 1H), 7.09 (m, 2H), 6.58 (t, J = 5.4 Hz, 1H), 6.01 (m, 1H), 5.95 (s, 1H), 5.57 (m, 1H), 5.06 (m, 1H), 4.09 (t, J = 9.1 Hz, 1H), 3.57 (d, J = 5.2 Hz, 2H), 3.44 (m, 3H), 3.30-2.53 (m, 8H), 2.14 (m, 7H), 1.48 (s, 3H), 1.25 (d, J = 5.5 Hz, 1H), 1.00 (m, 1H)	621.30
K12	11.09 (s, 1H), 8.11 (s, 1H), 7.59 (m, 1H), 7.15 (d, J = 8.6 Hz, 1H), 7.05 (d, J = 7.0 Hz, 1H), 6.02 (d, J = 3.4 Hz, 1H), 5.95 (t, J = 8.4 Hz, 1H), 5.57 (d, J = 3.1 Hz, 1H), 5.06 (m, 1H), 4.08 (t, J = 9.4 Hz, 1H), 3.60 (t, J = 5.4 Hz, 2H), 3.54-3.38 (m, 9H), 3.22-2.54 (m, 7H), 2.37-2.00 (m, 8H), 1.47 (s, 3H), 1.02 (t, J = 11.8 Hz, 1H)	664.51
K13	11.09 (s, 1H), 8.12 (t, J = 5.6 Hz, 1H), 7.59 (m, 1H), 7.15 (d, J = 8.6 Hz, 1H), 7.05 (d, J = 7.0 Hz, 1H), 6.02 (d, J = 3.4 Hz, 1H), 5.95 (t, J = 8.0 Hz, 1H), 5.76 (s, 1H), 5.57 (d, J = 3.1 Hz, 1H), 5.06 (m, 1H), 4.08 (t, J = 9.4 Hz, 1H), 3.65-3.36 (m, 15H), 2.77-2.57 (m, 3H), 2.37-1.52 (m, 10H), 1.47 (s, 3H), 1.28-1.21 (m, 2H)	709.40
K14	11.09 (s, 1H), 8.13 (t, J = 5.5 Hz, 1H), 7.59 (m, 1H), 7.15 (d, J = 8.6 Hz, 1H), 7.05 (d, J = 7.0 Hz, 1H), 6.04-5.89 (m, 2H), 5.57 (d, J = 3.1 Hz, 1H), 5.08-5.00 (m, 1H), 4.08 (t, J = 9.4 Hz, 1H), 3.63 (t, J = 5.4 Hz, 2H), 3.57-3.46 (m, 10H), 3.44 (s, 3H), 3.23-2.56 (m, 8H), 2.37-1.99 (m, 10H), 1.47 (s, 3H), 1.25 (d, J = 9.5 Hz, 2H)	752.49
K15	11.07 (s, 1H), 7.57 (m, 1H), 7.13 (t, J = 8.3 Hz, 2H), 6.03 (d, J = 3.5 Hz, 1H), 5.67-5.56 (m, 2H), 5.06 (m, 1H), 4.14 (t, J = 9.4 Hz, 1H), 3.55 (m, 6H), 2.88 (m, 3H), 2.74 (s, 1H), 2.34 (m, 3H), 2.22-1.52 (m, 11H), 1.50 (s, 3H), 1.42-1.01 (m, 4H)	656.56
K16	11.10 (s, 1H), 7.59 (m, 1H), 7.04 (m, 2H), 6.49 (d, J = 6.3 Hz, 1H), 6.02 (d, J = 3.5 Hz, 1H), 5.59 (m, 2H), 5.06 (m, 1H), 4.14 (t, J = 9.4 Hz, 2H), 2.96-2.82 (m, 2H), 2.69-2.55 (m, 2H), 2.35 (m, 6H), 2.23-1.96 (m, 5H), 1.76 (t, J = 9.3 Hz, 3H), 1.63 (m, 4H), 1.50 (s, 2H), 1.39 (d, J = 7.4 Hz, 1H), 1.27-1.23 (m, 3H)	656.73
K17	11.08 (s, 1H), 7.68 (m, 1H), 7.33 (m, 2H), 6.03 (d, J = 2.4 Hz, 1H), 5.65-5.51 (m, 2H), 5.09 (m, 1H), 4.14 (t, J = 9.3 Hz, 1H), 3.74 (d, J = 11.6 Hz, 2H), 2.87-2.81 (m, 3H), 2.66-2.54 (m, 3H), 2.44-1.72 (m, 13H), 1.51 (s, 3H), 1.44-1.36 (m, 3H), 1.24 (s, 3H), 1.13-1.00 (m, 3H)	684.59
K18	11.10 (s, 1H), 8.00 (t, J = 5.6 Hz, 1H), 7.81 (m, 1H), 7.48 (m, 2H), 6.02 (d, J = 3.4 Hz, 1H), 5.92 (t, J = 8.0 Hz, 1H), 5.54 (d, J = 2.9 Hz, 1H), 5.09 (m, 1H), 4.20 (t, J = 6.4 Hz, 2H), 4.10 (t, J = 9.4 Hz, 1H), 3.22 (m, 1H), 3.16 (d, J = 9.6 Hz, 1H), 3.01-2.56 (m, 5H), 2.55-2.52 (m, 1H), 2.34-2.00 (m, 7H), 1.79-1.72 (m, 2H), 1.58 (t, J = 11.3 Hz, 3H), 1.45-1.26 (m, 9H), 1.02 (m, 2H)	662.40
K19	11.06 (s, 1H), 8.00 (t, J = 5.7 Hz, 1H), 7.56 (d, J = 8.4 Hz, 1H), 7.08 (t, J = 5.3 Hz, 1H), 6.95 (d, J = 1.8 Hz, 1H), 6.85 (m, 1H), 6.03 (d, J = 3.5 Hz, 1H), 5.92 (t, J = 7.9 Hz, 1H), 5.54 (d, J = 3.1 Hz, 1H), 5.03 (m, 1H), 4.10 (t, J = 9.4 Hz, 1H), 3.26-3.12 (m, 4H), 3.02-2.52 (m, 6H), 2.37-1.51 (m, 9H), 1.48 (s, 3H), 1.34 (m, 6H), 1.24 (s, 3H)	647.50
K20	11.06 (s, 1H), 7.99 (t, J = 5.7 Hz, 1H), 7.53 (m, 1H), 7.09 (t, J = 5.2 Hz, 1H), 6.95 (d, J = 1.7 Hz, 1H), 6.85 (m, 1H), 6.02 (d, J = 3.5 Hz, 1H), 5.92 (t, J = 8.0 Hz, 1H), 5.54 (d, J = 3.1 Hz, 1H), 5.03 (m, 1H), 4.10 (t, J = 9.4 Hz, 1H), 3.25-2.52 (m, 14H), 2.37-1.96 (m, 8H), 1.64-1.53 (m, 3H), 1.24 (s, 3H), 1.21-1.16 (m, 3H), 1.02 (m, 2H)	661.80

Compd.

¹H NMR (400 MHz, DMSO-d₆)

ESI-MS
(m/z) [M+H]⁺

11.16 (s, 1H), 9.69 (s, 1H), 8.51 (d, J = 8.4 Hz, 1H), 7.84 (t, J = 7.9 Hz, 1H), 7.61 (d, J = 7.2 Hz, 1H), 5.26-5.12 (m, 2H), 3.94 (t, J = 9.1 Hz, 1H), 2.77 (d, J = 9.1 Hz, 1H), 2.59 (m, 4H), 2.47 (d, J = 7.0 Hz, 2H), 2.25-1.56 (m, 17H), 1.32 (d, J = 15.5 Hz, 2H), 1.25 (d, J = 10.2 Hz, 6H), 1.20 (s, 3H), 1.12 (m, 2H)

675.46

11.15 (s, 1H), 9.70 (s, 1H), 8.48 (d, J = 8.4 Hz, 1H), 7.87-7.81 (m, 1H), 7.62 (d, J = 7.1 Hz, 1H), 5.22-5.07 (m, 2H), 3.96 (t, J = 9.1 Hz, 1H), 2.78 (d, J = 9.1 Hz, 1H), 2.61 (m, 5H), 2.43-1.92 (m, 17H), 1.63 (d, J = 9.3 Hz, 6H), 1.55-1.48 (m, 3H), 1.24 (s, 3H), 1.20 (s, 2H), 0.89-0.81 (m, 2H)

689.57

11.15 (s, 1H), 9.70 (s, 1H), 8.48 (d, J = 8.4 Hz, 1H), 7.92-7.80 (m, 1H), 7.62 (d, J = 7.2 Hz, 1H), 5.26-5.11 (m, 2H), 3.96 (t, J = 9.1 Hz, 1H), 2.96-2.85 (m, 1H), 2.78 (d, J = 9.1 Hz, 1H), 2.60 (m, 6H), 2.48-1.91 (m, 17H), 1.74-1.56 (m, 6H), 1.48 (s, 2H), 1.33 (m, 3H), 1.20 (s, 3H), 1.11 (m, 2H)

703.64

11.16 (s, 1H), 9.69 (s, 1H), 8.48 (d, J = 8.4 Hz, 1H), 7.89-7.78 (m, 1H), 7.62 (d, J = 7.0 Hz, 1H), 5.24-5.10 (m, 2H), 3.96 (t, J = 9.1 Hz, 1H), 2.79 (d, J = 9.1 Hz, 1H), 2.67-2.58 (m, 4H), 2.49-2.44 (m, 4H), 2.42-1.91 (m, 14H), 1.69-1.23 (m, 19H), 1.20 (s, 3H)

732.60

11.15 (s, 1H), 9.70 (s, 1H), 8.50 (d, J = 8.4 Hz, 1H), 7.90-7.79 (m, 1H), 7.62 (d, J = 7.3 Hz, 1H), 6.03 (d, J = 3.4 Hz, 1H), 5.65-5.53 (m, 2H), 5.15 (m, 1H), 4.14 (t, J = 9.4 Hz, 1H), 3.18 (s, 1H), 2.96-2.83 (m, 2H), 2.61 (m, 2H), 2.48-1.57 (m, 19H), 1.50 (s, 2H), 1.43-1.29 (m, 1H), 1.28-1.21 (m, 2H), 1.16-0.98 (m, 2H)

687.30

11.16 (s, 1H), 9.70 (s, 1H), 8.48 (t, J = 6.9 Hz, 1H), 7.84 (t, J = 7.9 Hz, 1H), 7.62 (d, J = 7.2 Hz, 1H), 6.68 (d, J = 8.3 Hz, 1H), 6.03 (d, J = 3.4 Hz, 1H), 5.57 (d, J = 3.0 Hz, 1H), 5.15 (m, 1H), 4.15 (t, J = 9.4 Hz, 1H), 2.37-1.98 (m, 14H), 1.64 (d, J = 7.9 Hz, 4H), 1.32 (d, J = 14.5 Hz, 6H), 1.25 (t, J = 6.6 Hz, 11H)

715.30

11.10 (s, 1H), 8.09 (t, J = 5.7 Hz, 1H), 7.64-7.52 (m, 1H), 7.08-6.99 (m, 2H), 6.66 (t, J = 6.0 Hz, 1H), 5.98 (t, J = 5.6 Hz, 2H), 5.54 (d, J = 3.0 Hz, 1H), 4.17 (m, 3H), 3.14 (m, 2H), 2.65-2.54 (m, 2H), 2.15 (m, 7H), 1.74-1.62 (m, 4H), 1.49 (s, 3H), 1.25 (m, 4H)

591.20

11.09 (s, 1H), 8.02 (s, 1H), 7.65-7.52 (m, 1H), 7.06 (m, 2H), 6.51 (t, J = 5.6 Hz, 1H), 6.02 (d, J = 3.4 Hz, 1H), 5.97-5.87 (m, 1H), 5.56 (d, J = 2.4 Hz, 1H), 5.05 (m, 1H), 4.11 (t, J = 9.4 Hz, 1H), 3.27-3.15 (m, 3H), 2.95-2.82 (m, 1H), 2.65-2.53 (m, 2H), 2.36-1.53 (m, 10H), 1.48 (s, 3H), 1.45-1.23 (m, 8H)

647.50

11.09 (s, 1H), 7.59 (m, 1H), 7.06 (m, 2H), 6.02 (d, J = 3.4 Hz, 1H), 5.92 (t, J = 7.9 Hz, 1H), 5.54 (d, J = 3.1 Hz, 1H), 5.05 (m, 1H), 4.10 (t, J = 9.4 Hz, 1H), 3.30-2.56 (m, 12H), 2.35-1.52 (m, 12H), 1.48 (s, 3H), 1.24 (s, 3H), 1.03 (t, J = 12.6 Hz, 1H), 0.88-0.80 (m, 2H)

646.74

K10

11.09 (s, 1H), 8.14-7.83 (m, 1H), 7.58 (m, 2H), 7.06 (m, 3H), 6.51 (d, J = 5.3 Hz, 1H), 6.14-5.80 (m, 3H), 5.76 (s, 1H), 5.53 (d, J = 3.1 Hz, 2H), 5.03 (m, 2H), 4.05 (m, 2H), 3.31-3.25 (m, 3H), 3.24-3.11 (m, 2H), 3.03-2.91 (m, 2H), 2.91-2.81 (m, 1H), 2.80-2.65 (m, 2H), 2.65-2.54 (m, 3H), 2.32 (d, J = 12.1 Hz, 2H), 2.24 (m, 3H), 2.22-2.14 (m, 2H), 2.10 (m, 2H), 2.04 (m, 1H), 1.59 (t, J = 15.4 Hz, 5H), 1.48 (s, 4H), 0.84 (m, 2H)

716.75

K11

11.10 (s, 1H), 8.12 (s, 1H), 7.60 (m, 1H), 7.09 (m, 2H), 6.58 (t, J = 5.4 Hz, 1H), 6.01 (m, 1H), 5.95 (s, 1H), 5.57 (m, 1H), 5.06 (m, 1H), 4.09 (t, J = 9.1 Hz, 1H), 3.57 (d, J = 5.2 Hz, 2H), 3.44 (m, 3H), 3.30-2.53 (m, 8H), 2.14 (m, 7H), 1.48 (s, 3H), 1.25 (d, J = 5.5 Hz, 1H), 1.00 (m, 1H)

621.30

K12

11.09 (s, 1H), 8.11 (s, 1H), 7.59 (m, 1H), 7.15 (d, J = 8.6 Hz, 1H), 7.05 (d, J = 7.0 Hz, 1H), 6.02 (d, J = 3.4 Hz, 1H), 5.95 (t, J = 8.4 Hz, 1H), 5.57 (d, J = 3.1 Hz, 1H), 5.06 (m, 1H), 4.08 (t, J = 9.4 Hz, 1H), 3.60 (t, J = 5.4 Hz, 2H), 3.54-3.38 (m, 9H), 3.22-2.54 (m, 7H), 2.37-2.00 (m, 8H), 1.47 (s, 3H), 1.02 (t, J = 11.8 Hz, 1H)

664.51

K13

11.09 (s, 1H), 8.12 (t, J = 5.6 Hz, 1H), 7.59 (m, 1H), 7.15 (d, J = 8.6 Hz, 1H), 7.05 (d, J = 7.0 Hz, 1H), 6.02 (d, J = 3.4 Hz, 1H), 5.95 (t, J = 8.0 Hz, 1H), 5.76 (s, 1H), 5.57 (d, J = 3.1 Hz, 1H), 5.06 (m, 1H), 4.08 (t, J = 9.4 Hz, 1H), 3.65-3.36 (m, 15H), 2.77-2.57 (m, 3H), 2.37-1.52 (m, 10H), 1.47 (s, 3H), 1.28-1.21 (m, 2H)

709.40

K14

11.09 (s, 1H), 8.13 (t, J = 5.5 Hz, 1H), 7.59 (m, 1H), 7.15 (d, J = 8.6 Hz, 1H), 7.05 (d, J = 7.0 Hz, 1H), 6.04-5.89 (m, 2H), 5.57 (d, J = 3.1 Hz, 1H), 5.08-5.00 (m, 1H), 4.08 (t, J = 9.4 Hz, 1H), 3.63 (t, J = 5.4 Hz, 2H), 3.57-3.46 (m, 10H), 3.44 (s, 3H), 3.23-2.56 (m, 8H), 2.37-1.99 (m, 10H), 1.47 (s, 3H), 1.25 (d, J = 9.5 Hz, 2H)

752.49

K15

11.07 (s, 1H), 7.57 (m, 1H), 7.13 (t, J = 8.3 Hz, 2H), 6.03 (d, J = 3.5 Hz, 1H), 5.67-5.56 (m, 2H), 5.06 (m, 1H), 4.14 (t, J = 9.4 Hz, 1H), 3.55 (m, 6H), 2.88 (m, 3H), 2.74 (s, 1H), 2.34 (m, 3H), 2.22-1.52 (m, 11H), 1.50 (s, 3H), 1.42-1.01 (m, 4H)

656.56

K16

11.10 (s, 1H), 7.59 (m, 1H), 7.04 (m, 2H), 6.49 (d, J = 6.3 Hz, 1H), 6.02 (d, J = 3.5 Hz, 1H), 5.59 (m, 2H), 5.06 (m, 1H), 4.14 (t, J = 9.4 Hz, 2H), 2.96-2.82 (m, 2H), 2.69-2.55 (m, 2H), 2.35 (m, 6H), 2.23-1.96 (m, 5H), 1.76 (t, J = 9.3 Hz, 3H), 1.63 (m, 4H), 1.50 (s, 2H), 1.39 (d, J = 7.4 Hz, 1H), 1.27-1.23 (m, 3H)

656.73

K17

11.08 (s, 1H), 7.68 (m, 1H), 7.33 (m, 2H), 6.03 (d, J = 2.4 Hz, 1H), 5.65-5.51 (m, 2H), 5.09 (m, 1H), 4.14 (t, J = 9.3 Hz, 1H), 3.74 (d, J = 11.6 Hz, 2H), 2.87-2.81 (m, 3H), 2.66-2.54 (m, 3H), 2.44-1.72 (m, 13H), 1.51 (s, 3H), 1.44-1.36 (m, 3H), 1.24 (s, 3H), 1.13-1.00 (m, 3H)

684.59

K18

11.10 (s, 1H), 8.00 (t, J = 5.6 Hz, 1H), 7.81 (m, 1H), 7.48 (m, 2H), 6.02 (d, J = 3.4 Hz, 1H), 5.92 (t, J = 8.0 Hz, 1H), 5.54 (d, J = 2.9 Hz, 1H), 5.09 (m, 1H), 4.20 (t, J = 6.4 Hz, 2H), 4.10 (t, J = 9.4 Hz, 1H), 3.22 (m, 1H), 3.16 (d, J = 9.6 Hz, 1H), 3.01-2.56 (m, 5H), 2.55-2.52 (m, 1H), 2.34-2.00 (m, 7H), 1.79-1.72 (m, 2H), 1.58 (t, J = 11.3 Hz, 3H), 1.45-1.26 (m, 9H), 1.02 (m, 2H)

662.40

K19

11.06 (s, 1H), 8.00 (t, J = 5.7 Hz, 1H), 7.56 (d, J = 8.4 Hz, 1H), 7.08 (t, J = 5.3 Hz, 1H), 6.95 (d, J = 1.8 Hz, 1H), 6.85 (m, 1H), 6.03 (d, J = 3.5 Hz, 1H), 5.92 (t, J = 7.9 Hz, 1H), 5.54 (d, J = 3.1 Hz, 1H), 5.03 (m, 1H), 4.10 (t, J = 9.4 Hz, 1H), 3.26-3.12 (m, 4H), 3.02-2.52 (m, 6H), 2.37-1.51 (m, 9H), 1.48 (s, 3H), 1.34 (m, 6H), 1.24 (s, 3H)

647.50

K20

11.06 (s, 1H), 7.99 (t, J = 5.7 Hz, 1H), 7.53 (m, 1H), 7.09 (t, J = 5.2 Hz, 1H), 6.95 (d, J = 1.7 Hz, 1H), 6.85 (m, 1H), 6.02 (d, J = 3.5 Hz, 1H), 5.92 (t, J = 8.0 Hz, 1H), 5.54 (d, J = 3.1 Hz, 1H), 5.03 (m, 1H), 4.10 (t, J = 9.4 Hz, 1H), 3.25-2.52 (m, 14H), 2.37-1.96 (m, 8H), 1.64-1.53 (m, 3H), 1.24 (s, 3H), 1.21-1.16 (m, 3H), 1.02 (m, 2H)

661.80


K1		n = 3	1	6.45	10.60
K2		n = 4	1	8.85	14.41
K3		n = 5	1	19.09	19.25
K4		n = 7	1	18.83	34.13
K5		n = 3	1	N.D^b.	N.D.
K6		n = 5	1	N.D.	N.D.
K7		n = 3	1	N.D.	N.D.
K8		n = 5	1	26.16	30.63
K9		n = 7	1	2.81	4.67
K10		n = 12	1	2.03	2.50
K11		n = 1	1	38.29	N.D.
K12		n = 2	1	11.67	N.D.
K13		n = 3	1	41.67	N.D.
K14		n = 4	1	N.D.	N.D.
K15			1	4.57	9.89
K16			1	4.29	6.63
K17			1	2.11	4.60
K18		n = 8	1	3.63	5.85
K19		n = 7	2	33.81	38.06
K20		n = 8	2	5.18	8.01
FT671	--		--	8.32	N.D.

Compd.

Linker

Site

IC₅₀/μmol·L^-1 (mean ± SD) (n = 2)^a

HCT-116

HT29

n = 3

6.45

10.60

n = 4

8.85

14.41

n = 5

19.09

19.25

n = 7

18.83

34.13

n = 3

N.D^b.

N.D.

n = 5

N.D.

n = 3

N.D.

n = 5

26.16

30.63

n = 7

2.81

4.67

K10

n = 12

2.03

2.50

K11

n = 1

38.29

N.D.

K12

n = 2

11.67

N.D.

K13

n = 3

41.67

N.D.

K14

n = 4

N.D.

K15

4.57

9.89

K16

4.29

6.63

K17

2.11

4.60

K18

n = 8

3.63

5.85

K19

n = 7

33.81

38.06

K20

n = 8

5.18

8.01

FT671

8.32

N.D.