药学学报

Compd.	mp, ¹H NMR, ¹³C NMR, HRMS-ESI
Ⅰ-1	mp: 159-162 ℃. ¹H NMR (500 MHz, CDCl₃), δ: 9.72 (s, 1H, ArH), 8.54 (d, J = 5.3 Hz, 1H, ArH), 7.50 (s, 1H, ArH), 7.46 (d, J = 5.3 Hz, 1H, ArH), 3.78 (p, J = 8.4 Hz, 1H, -CH), 2.61-2.53 (m, 2H, CH₂), 2.45-2.38 (m, 2H, CH₂), 2.19-2.00 (m, 2H, CH₂); ¹³C NMR (125 MHz, CDCl₃), δ: 172.0, 143.5, 138.7, 134.9, 132.0, 126.5, 113.9, 98.4, 39.0, 25.2, 18.3. HRMS: (ESI, m/z); [M+H]⁺ calcd for 279.012 8; found 279.013 8.
Ⅰ-2	mp: 102-105 ℃. ¹H NMR (500 MHz, CDCl₃), δ: 9.64 (s, 1H, ArH), 8.49 (d, J = 5.3 Hz, 1H, ArH), 7.77-7.74 (m, 2H, ArH), 7.67-7.63 (m, 1H, ArH), 7.57-7.54 (m, 3H, ArH), 7.49 (d, J = 3.7 Hz, 1H, ArH), 6.64 (d, J = 3.5 Hz, 1H, ArH); ¹³C NMR (125 MHz, CDCl₃), δ: 168.4, 143.4, 138.8, 136.6, 133.9, 133.4, 132.9, 131.0, 129.6, 129.2, 115.9, 107.8. HRMS: (ESI, m/z); [M+H]⁺ calcd for 223.086 6; found 223.086 9.
Ⅰ-3	mp: 119-121 ℃。¹H NMR (500 MHz, CDCl₃), δ: 9.25 (s, 1H, ArH), 8.56 (d, J = 5.5 Hz, 1H, ArH), 7.61 (s, 1H, ArH), 7.55-7.53(m, 1H, ArH), 2.67-2.62 (m, 1H, CH), 1.48-1.44 (m, 2H, CH₂), 1.15-1.10 (m, 2H, CH₂); ¹³C NMR (125 MHz, CDCl₃), δ: 142.7, 135.7, 135.2, 131.5, 128.1, 114.2, 97.3, 32.0, 6.6. HRMS: (ESI, m/z); [M+H]⁺ calcd for 300.964 1; found 300.963 5.
Ⅱ-1	mp: 215-217 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.74 (s, 1H, ArH), 8.15-8.14 (m, 1H, ArH), 7.87 (d, J = 5.4 Hz, 1H, ArH), 7.79-7.78 (m, 1H, ArH), 7.50-7.46 (m, 2H, ArH), 7.34-7.31 (m, 1H, ArH), 7.11 (d, J = 7.5 Hz, 1H, ArH), 2.42 (s, 3H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆) δ: 138.5, 137.9, 135.0, 134.7, 129.0, 128.8, 127.2, 127.0, 126.4, 124.1, 115.2, 113.8, 113.4, 21.7. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 209.107 3; found: 209.108 4.
Ⅱ-2	mp: 214-217 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.12 (s, 1H, NH), 9.35 (d, J = 2.3 Hz, 1H, ArH), 8.86 (s, 1H, ArH), 8.67 (d, J = 2.6 Hz, 1H, ArH), 8.33 (d, J = 2.4 Hz, 1H, ArH), 8.27 (d, J = 5.5 Hz, 1H, ArH), 8.08 (t, J = 6.2 Hz, 2H, ArH), 8.02 (d, J = 8.2 Hz, 1H, ArH), 7.72-7.69 (m, 1H, ArH), 7.64-7.60 (m, 1H, ArH); ¹³C NMR (125 MHz, MeOD-d₄), δ: 150.6, 147.0, 139.1, 135.7, 135.5, 133.8, 131.8, 130.3, 130.0, 129.9, 129.7, 129.1, 128.9, 128.4, 115.3, 114.0. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 246.102 6; found: 246.102 2.
Ⅱ-3	mp: 211-213 ℃. ¹H NMR (500 MHz, MeOD-d₄) δ: 8.74 (s, 1H, ArH), 8.14 (d, J = 5.7 Hz, 1H, ArH), 7.89-7.87(m, 1H, ArH), 7.80 (s, 1H, ArH), 7.69-7.67 (m, 2H, ArH), 7.44 (t, J = 7.7 Hz, 2H, ArH), 7.31-7.26 (m, 1H, ArH). HRMS: (ESI, m/z); [M+H]⁺ calcd for: 195.091 7; found: 195.091 2.
Ⅱ-4	mp: 218-220 ℃. ¹H NMR (500 MHz, MeOD-d₄) δ: 8.71 (s, 1H, ArH), 8.08 (d, J = 5.6 Hz, 1H, ArH), 7.79 (s, 1H, ArH), 7.68 (d, J = 5.6 Hz, 1H, ArH), 7.55 (d, J = 9.3 Hz, 1H, ArH), 7.31-7.27 (m, 1H, ArH), 7.10 (d, J = 9.5 Hz, 1H, ArH), 7.06-7.02 (m, 1H, ArH), 3.85 (s, 3H, OCH₃); ¹³C NMR (125 MHz, DMSO-d₆) δ: 156.1, 137.7, 134.5, 133.3, 130.3, 129.5, 129.2, 127.4, 123.0, 120.6, 114.5, 111.6, 111.5, 55.3. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 225.102 2; found: 225.101 7.
Ⅲ-1	mp: 214-216 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.36 (s, 1H, NH), 8.83 (d, J = 1.1 Hz, 1H, ArH), 8.28 (d, J = 5.5 Hz, 1H, ArH), 7.94 (d, J = 5.5 Hz, 1H, ArH), 7.88 (s, 1H, ArH), 7.75 (s, 1H, ArH), 2.48 (s, 3H, CH₃), 2.32 (s, 3H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 206.7, 197.7, 140.0, 137.9, 135.80, 133.8, 132.5, 132.4, 132.0, 113.9, 109.0, 31.4, 26.5. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 229.097 2; found: 229.097 1.
Ⅲ-2	mp: 165-168 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.27 (s, 1H, NH), 8.82 (d, J = 1.1 Hz, 1H, ArH), 8.26 (d, J = 5.5 Hz, 1H, ArH), 7.92 (d, J = 5.3 Hz, 2H, ArH), 7.60 (s, 1H, ArH), 2.92 (q, J = 7.2 Hz, 2H, CH₂), 2.58 (q, J = 7.2 Hz, 2H, CH₂), 1.06-1.02 (m, 6H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 209.2, 199.6, 139.4, 136.9, 135.3, 133.3, 131.8, 131.3, 130.3, 113.4, 108.6, 36.0, 30.2, 8.4, 7.7. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 257.128 5; found: 257.129 1.
Ⅲ-3	mp: 180-181 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.41 (s, 1H, NH), 8.83 (d, J = 1.1 Hz, 1H, ArH), 8.25 (d, J = 5.5 Hz, 1H, ArH), 7.99 (s, 1H, ArH), 7.94 (s, 1H, ArH), 7.72-7.71 (m, 1H, ArH), 3.84 (s, 3H, OCH₃), 3.77 (s, 3H, OCH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 167.6, 164.9, 140.0, 135.6, 134.5, 133.6, 133.1, 131.6, 118.9, 113.2, 108.4, 52.8, 52.4. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 261.087 0; found: 261.087 0.
Ⅲ-4	mp: 171-173 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.75 (s, 1H, ArH), 8.23 (d, J = 5.7 Hz, 1H, ArH), 8.04–7.99 (m, 2H, ArH), 7.77-7.76 (m, 1H, ArH), 4.38 (q, J = 7.1 Hz, 2H, OCH₂), 4.31 (q, J = 7.1 Hz, 2H, OCH₂), 1.36-1.32 (m, 6H, CH₃); ¹³C NMR (125 MHz, MeOD-d₄), δ: 169.1, 166.7, 140.0, 135.7, 135.0, 134.9, 134.5, 134.2, 122.2, 114.7, 110.6, 62.9, 62.6, 14.7, 14.5. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 289.118 3; found: 289.118 6.
Ⅲ-5	mp: 163-164 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.38 (s, 1H, NH), 8.83 (d, J = 1.1 Hz, 1H, ArH), 8.25 (d, J = 5.6 Hz, 1H, ArH), 7.95 (d, J = 1.1 Hz, 2H, ArH), 7.72-7.71 (m, 1H, ArH), 4.24 (t, J = 6.5 Hz, 2H, OCH₂), 4.14 (t, J = 6.5 Hz, 2H, OCH₂), 1.67-1.62 (m, 4H, CH₂), 0.92 (t, J = 7.4 Hz, 3H, CH₃), 0.87 (t, J = 7.4 Hz, 3H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 167.1, 164.3, 139.7, 135.4, 133.7, 133.4, 132.7, 131.4, 119.6, 113.0, 108.3, 66.7, 66.2, 21.6, 21.3, 10.2. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 317.149 6; found: 317.150 4.
Ⅳ-1	mp: 129-135 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.64 (d, J = 1.1 Hz, 1H, ArH), 8.04 (d, J = 5.6 Hz, 1H, ArH), 7.66-7.65 (m, 1H, ArH), 7.50 (s, 1H, ArH), 7.11-7.07 (m, 2H, ArH), 6.73-6.71 (m, 2H, ArH), 6.63-6.60 (m, 1H, ArH), 4.45 (s, 2H, CH₂); ¹³C NMR (125 MHz, MeOD-d₄), δ: 150.4, 137.7, 135.4, 134.7, 133.5, 130.1, 129.7, 118.4, 115.3, 115.0, 114.5, 40.4. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 224.118 2; found: 224.119 0.
Ⅳ-2	mp: 155-156 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.64 (s, 1H, ArH), 8.04 (d, J = 5.6 Hz, 1H, ArH), 7.65-7.63 (m, 1H, ArH), 7.49 (s 1H, ArH), 7.05-7.02 (m, 2H, ArH), 6.66-6.63 (m, 2H, ArH), 4.42 (s, 2H, CH₂); ¹³C NMR (125 MHz, MeOD-d₄), δ: 149.0, 137.6, 135.3, 134.6, 133.3, 129.7, 129.6, 122.3, 117.5, 115.2, 114.5, 40.2. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 258.079 3; found: 258.080 1.
Ⅳ-3	mp: 156-160 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.60 (s, 1H, ArH), 8.00 (d, J = 5.6 Hz, 1H), 7.61-7.59 (m, 1H, ArH), 7.45 (s, 1H, ArH), 7.14-7.11 (m, 2H, ArH), 6.58–6.55 (m, 2H, ArH), 4.38 (s, 2H, CH₂); ¹³C NMR (125 MHz, MeOD-d₄), δ: 149.4, 137.6, 135.3, 134.7, 133.3, 132.6, 129.6, 115.7, 115.1, 114.4, 109.1, 40.1. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 302.028 7; found: 302.029 0.
Ⅳ-4	mp: 143-145 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 8.72 (s, 1H, ArH), 8.07 (d, J = 5.5 Hz, 1H, ArH), 7.63 (d, J = 5.5 Hz, 1H, ArH), 7.54 (s, 1H, ArH), 6.85 (d, J = 8.0 Hz, 2H, ArH), 6.58 (d, J = 8.1 Hz, 2H, ArH), 4.34 (s, 2H, CH₂), 2.11 (s, 3H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 146.7, 137.3, 134.4, 133.6, 131.1, 129.4, 127.8, 124.4, 113.9, 113.2, 112.8, 38.7, 20.2. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 238.133 9; found: 238.134 7.
Ⅳ-5	mp: 146-148 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 8.71 (s, 1H, ArH), 8.07 (d, J = 5.5 Hz, 1H, ArH), 7.63 (d, J = 5.5 Hz, 1H, ArH), 7.54 (s, 1H, ArH), 6.68 (d, J = 8.9 Hz, 2H, ArH), 6.62 (d, J = 8.7 Hz, 2H, ArH), 4.32 (s, 2H, CH₂), 3.63 (s, 3H, OCH₃); ¹³C NMR (125 MHz, MeOD-d₄), δ: 153.9, 144.2, 137.5, 135.2, 134.5, 133.4, 129.8, 116.3, 115.7, 115.2, 114.9, 56.2, 41.5. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 254.128 8; found: 254.129 6.
Ⅳ-6	mp: 134-136 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 8.71 (s, 1H), 8.07 (d, J = 5.5 Hz, 1H, ArH), 7.63 (d, J = 5.5 Hz, 1H, ArH), 7.55 (s, 1H, ArH), 6.87 (t, J = 8.7 Hz, 2H, ArH), 6.65-6.63 (m, 2H, ArH), 4.34 (s, 2H, CH₂); ¹³C NMR (125 MHz, DMSO-d₆), δ: 154.5 (d, J = 230.8 Hz), 145.7, 137.4, 134.5, 133.6, 131.1, 127.9, 115.3 (d, J = 21.8 Hz), 113.9, 113.3 (d, J = 7.2 Hz), 112.8, 38.9. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 242.108 8; found: 242.109 1.

Compd.

mp, ¹H NMR, ¹³C NMR, HRMS-ESI

Ⅰ-1

mp: 159-162 ℃. ¹H NMR (500 MHz, CDCl₃), δ: 9.72 (s, 1H, ArH), 8.54 (d, J = 5.3 Hz, 1H, ArH), 7.50 (s, 1H, ArH), 7.46 (d, J = 5.3 Hz, 1H, ArH), 3.78 (p, J = 8.4 Hz, 1H, -CH), 2.61-2.53 (m, 2H, CH₂), 2.45-2.38 (m, 2H, CH₂), 2.19-2.00 (m, 2H, CH₂); ¹³C NMR (125 MHz, CDCl₃), δ: 172.0, 143.5, 138.7, 134.9, 132.0, 126.5, 113.9, 98.4, 39.0, 25.2, 18.3. HRMS: (ESI, m/z); [M+H]⁺ calcd for 279.012 8; found 279.013 8.

Ⅰ-2

mp: 102-105 ℃. ¹H NMR (500 MHz, CDCl₃), δ: 9.64 (s, 1H, ArH), 8.49 (d, J = 5.3 Hz, 1H, ArH), 7.77-7.74 (m, 2H, ArH), 7.67-7.63 (m, 1H, ArH), 7.57-7.54 (m, 3H, ArH), 7.49 (d, J = 3.7 Hz, 1H, ArH), 6.64 (d, J = 3.5 Hz, 1H, ArH); ¹³C NMR (125 MHz, CDCl₃), δ: 168.4, 143.4, 138.8, 136.6, 133.9, 133.4, 132.9, 131.0, 129.6, 129.2, 115.9, 107.8. HRMS: (ESI, m/z); [M+H]⁺ calcd for 223.086 6; found 223.086 9.

Ⅰ-3

mp: 119-121 ℃。¹H NMR (500 MHz, CDCl₃), δ: 9.25 (s, 1H, ArH), 8.56 (d, J = 5.5 Hz, 1H, ArH), 7.61 (s, 1H, ArH), 7.55-7.53(m, 1H, ArH), 2.67-2.62 (m, 1H, CH), 1.48-1.44 (m, 2H, CH₂), 1.15-1.10 (m, 2H, CH₂); ¹³C NMR (125 MHz, CDCl₃), δ: 142.7, 135.7, 135.2, 131.5, 128.1, 114.2, 97.3, 32.0, 6.6. HRMS: (ESI, m/z); [M+H]⁺ calcd for 300.964 1; found 300.963 5.

Ⅱ-1

mp: 215-217 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.74 (s, 1H, ArH), 8.15-8.14 (m, 1H, ArH), 7.87 (d, J = 5.4 Hz, 1H, ArH), 7.79-7.78 (m, 1H, ArH), 7.50-7.46 (m, 2H, ArH), 7.34-7.31 (m, 1H, ArH), 7.11 (d, J = 7.5 Hz, 1H, ArH), 2.42 (s, 3H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆) δ: 138.5, 137.9, 135.0, 134.7, 129.0, 128.8, 127.2, 127.0, 126.4, 124.1, 115.2, 113.8, 113.4, 21.7. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 209.107 3; found: 209.108 4.

Ⅱ-2

mp: 214-217 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.12 (s, 1H, NH), 9.35 (d, J = 2.3 Hz, 1H, ArH), 8.86 (s, 1H, ArH), 8.67 (d, J = 2.6 Hz, 1H, ArH), 8.33 (d, J = 2.4 Hz, 1H, ArH), 8.27 (d, J = 5.5 Hz, 1H, ArH), 8.08 (t, J = 6.2 Hz, 2H, ArH), 8.02 (d, J = 8.2 Hz, 1H, ArH), 7.72-7.69 (m, 1H, ArH), 7.64-7.60 (m, 1H, ArH); ¹³C NMR (125 MHz, MeOD-d₄), δ: 150.6, 147.0, 139.1, 135.7, 135.5, 133.8, 131.8, 130.3, 130.0, 129.9, 129.7, 129.1, 128.9, 128.4, 115.3, 114.0. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 246.102 6; found: 246.102 2.

Ⅱ-3

mp: 211-213 ℃. ¹H NMR (500 MHz, MeOD-d₄) δ: 8.74 (s, 1H, ArH), 8.14 (d, J = 5.7 Hz, 1H, ArH), 7.89-7.87(m, 1H, ArH), 7.80 (s, 1H, ArH), 7.69-7.67 (m, 2H, ArH), 7.44 (t, J = 7.7 Hz, 2H, ArH), 7.31-7.26 (m, 1H, ArH). HRMS: (ESI, m/z); [M+H]⁺ calcd for: 195.091 7; found: 195.091 2.

Ⅱ-4

mp: 218-220 ℃. ¹H NMR (500 MHz, MeOD-d₄) δ: 8.71 (s, 1H, ArH), 8.08 (d, J = 5.6 Hz, 1H, ArH), 7.79 (s, 1H, ArH), 7.68 (d, J = 5.6 Hz, 1H, ArH), 7.55 (d, J = 9.3 Hz, 1H, ArH), 7.31-7.27 (m, 1H, ArH), 7.10 (d, J = 9.5 Hz, 1H, ArH), 7.06-7.02 (m, 1H, ArH), 3.85 (s, 3H, OCH₃); ¹³C NMR (125 MHz, DMSO-d₆) δ: 156.1, 137.7, 134.5, 133.3, 130.3, 129.5, 129.2, 127.4, 123.0, 120.6, 114.5, 111.6, 111.5, 55.3. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 225.102 2; found: 225.101 7.

Ⅲ-1

mp: 214-216 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.36 (s, 1H, NH), 8.83 (d, J = 1.1 Hz, 1H, ArH), 8.28 (d, J = 5.5 Hz, 1H, ArH), 7.94 (d, J = 5.5 Hz, 1H, ArH), 7.88 (s, 1H, ArH), 7.75 (s, 1H, ArH), 2.48 (s, 3H, CH₃), 2.32 (s, 3H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 206.7, 197.7, 140.0, 137.9, 135.80, 133.8, 132.5, 132.4, 132.0, 113.9, 109.0, 31.4, 26.5. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 229.097 2; found: 229.097 1.

Ⅲ-2

mp: 165-168 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.27 (s, 1H, NH), 8.82 (d, J = 1.1 Hz, 1H, ArH), 8.26 (d, J = 5.5 Hz, 1H, ArH), 7.92 (d, J = 5.3 Hz, 2H, ArH), 7.60 (s, 1H, ArH), 2.92 (q, J = 7.2 Hz, 2H, CH₂), 2.58 (q, J = 7.2 Hz, 2H, CH₂), 1.06-1.02 (m, 6H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 209.2, 199.6, 139.4, 136.9, 135.3, 133.3, 131.8, 131.3, 130.3, 113.4, 108.6, 36.0, 30.2, 8.4, 7.7. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 257.128 5; found: 257.129 1.

Ⅲ-3

mp: 180-181 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.41 (s, 1H, NH), 8.83 (d, J = 1.1 Hz, 1H, ArH), 8.25 (d, J = 5.5 Hz, 1H, ArH), 7.99 (s, 1H, ArH), 7.94 (s, 1H, ArH), 7.72-7.71 (m, 1H, ArH), 3.84 (s, 3H, OCH₃), 3.77 (s, 3H, OCH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 167.6, 164.9, 140.0, 135.6, 134.5, 133.6, 133.1, 131.6, 118.9, 113.2, 108.4, 52.8, 52.4. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 261.087 0; found: 261.087 0.

Ⅲ-4

mp: 171-173 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.75 (s, 1H, ArH), 8.23 (d, J = 5.7 Hz, 1H, ArH), 8.04–7.99 (m, 2H, ArH), 7.77-7.76 (m, 1H, ArH), 4.38 (q, J = 7.1 Hz, 2H, OCH₂), 4.31 (q, J = 7.1 Hz, 2H, OCH₂), 1.36-1.32 (m, 6H, CH₃); ¹³C NMR (125 MHz, MeOD-d₄), δ: 169.1, 166.7, 140.0, 135.7, 135.0, 134.9, 134.5, 134.2, 122.2, 114.7, 110.6, 62.9, 62.6, 14.7, 14.5. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 289.118 3; found: 289.118 6.

Ⅲ-5

mp: 163-164 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.38 (s, 1H, NH), 8.83 (d, J = 1.1 Hz, 1H, ArH), 8.25 (d, J = 5.6 Hz, 1H, ArH), 7.95 (d, J = 1.1 Hz, 2H, ArH), 7.72-7.71 (m, 1H, ArH), 4.24 (t, J = 6.5 Hz, 2H, OCH₂), 4.14 (t, J = 6.5 Hz, 2H, OCH₂), 1.67-1.62 (m, 4H, CH₂), 0.92 (t, J = 7.4 Hz, 3H, CH₃), 0.87 (t, J = 7.4 Hz, 3H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 167.1, 164.3, 139.7, 135.4, 133.7, 133.4, 132.7, 131.4, 119.6, 113.0, 108.3, 66.7, 66.2, 21.6, 21.3, 10.2. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 317.149 6; found: 317.150 4.

Ⅳ-1

mp: 129-135 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.64 (d, J = 1.1 Hz, 1H, ArH), 8.04 (d, J = 5.6 Hz, 1H, ArH), 7.66-7.65 (m, 1H, ArH), 7.50 (s, 1H, ArH), 7.11-7.07 (m, 2H, ArH), 6.73-6.71 (m, 2H, ArH), 6.63-6.60 (m, 1H, ArH), 4.45 (s, 2H, CH₂); ¹³C NMR (125 MHz, MeOD-d₄), δ: 150.4, 137.7, 135.4, 134.7, 133.5, 130.1, 129.7, 118.4, 115.3, 115.0, 114.5, 40.4. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 224.118 2; found: 224.119 0.

Ⅳ-2

mp: 155-156 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.64 (s, 1H, ArH), 8.04 (d, J = 5.6 Hz, 1H, ArH), 7.65-7.63 (m, 1H, ArH), 7.49 (s 1H, ArH), 7.05-7.02 (m, 2H, ArH), 6.66-6.63 (m, 2H, ArH), 4.42 (s, 2H, CH₂); ¹³C NMR (125 MHz, MeOD-d₄), δ: 149.0, 137.6, 135.3, 134.6, 133.3, 129.7, 129.6, 122.3, 117.5, 115.2, 114.5, 40.2. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 258.079 3; found: 258.080 1.

Ⅳ-3

mp: 156-160 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.60 (s, 1H, ArH), 8.00 (d, J = 5.6 Hz, 1H), 7.61-7.59 (m, 1H, ArH), 7.45 (s, 1H, ArH), 7.14-7.11 (m, 2H, ArH), 6.58–6.55 (m, 2H, ArH), 4.38 (s, 2H, CH₂); ¹³C NMR (125 MHz, MeOD-d₄), δ: 149.4, 137.6, 135.3, 134.7, 133.3, 132.6, 129.6, 115.7, 115.1, 114.4, 109.1, 40.1. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 302.028 7; found: 302.029 0.

Ⅳ-4

mp: 143-145 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 8.72 (s, 1H, ArH), 8.07 (d, J = 5.5 Hz, 1H, ArH), 7.63 (d, J = 5.5 Hz, 1H, ArH), 7.54 (s, 1H, ArH), 6.85 (d, J = 8.0 Hz, 2H, ArH), 6.58 (d, J = 8.1 Hz, 2H, ArH), 4.34 (s, 2H, CH₂), 2.11 (s, 3H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 146.7, 137.3, 134.4, 133.6, 131.1, 129.4, 127.8, 124.4, 113.9, 113.2, 112.8, 38.7, 20.2. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 238.133 9; found: 238.134 7.

Ⅳ-5

mp: 146-148 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 8.71 (s, 1H, ArH), 8.07 (d, J = 5.5 Hz, 1H, ArH), 7.63 (d, J = 5.5 Hz, 1H, ArH), 7.54 (s, 1H, ArH), 6.68 (d, J = 8.9 Hz, 2H, ArH), 6.62 (d, J = 8.7 Hz, 2H, ArH), 4.32 (s, 2H, CH₂), 3.63 (s, 3H, OCH₃); ¹³C NMR (125 MHz, MeOD-d₄), δ: 153.9, 144.2, 137.5, 135.2, 134.5, 133.4, 129.8, 116.3, 115.7, 115.2, 114.9, 56.2, 41.5. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 254.128 8; found: 254.129 6.

Ⅳ-6

mp: 134-136 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 8.71 (s, 1H), 8.07 (d, J = 5.5 Hz, 1H, ArH), 7.63 (d, J = 5.5 Hz, 1H, ArH), 7.55 (s, 1H, ArH), 6.87 (t, J = 8.7 Hz, 2H, ArH), 6.65-6.63 (m, 2H, ArH), 4.34 (s, 2H, CH₂); ¹³C NMR (125 MHz, DMSO-d₆), δ: 154.5 (d, J = 230.8 Hz), 145.7, 137.4, 134.5, 133.6, 131.1, 127.9, 115.3 (d, J = 21.8 Hz), 113.9, 113.3 (d, J = 7.2 Hz), 112.8, 38.9. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 242.108 8; found: 242.109 1.

Compd.	mp, ¹H NMR, ¹³C NMR, HRMS-ESI
Ⅰ-1	mp: 159-162 ℃. ¹H NMR (500 MHz, CDCl₃), δ: 9.72 (s, 1H, ArH), 8.54 (d, J = 5.3 Hz, 1H, ArH), 7.50 (s, 1H, ArH), 7.46 (d, J = 5.3 Hz, 1H, ArH), 3.78 (p, J = 8.4 Hz, 1H, -CH), 2.61-2.53 (m, 2H, CH₂), 2.45-2.38 (m, 2H, CH₂), 2.19-2.00 (m, 2H, CH₂); ¹³C NMR (125 MHz, CDCl₃), δ: 172.0, 143.5, 138.7, 134.9, 132.0, 126.5, 113.9, 98.4, 39.0, 25.2, 18.3. HRMS: (ESI, m/z); [M+H]⁺ calcd for 279.012 8; found 279.013 8.
Ⅰ-2	mp: 102-105 ℃. ¹H NMR (500 MHz, CDCl₃), δ: 9.64 (s, 1H, ArH), 8.49 (d, J = 5.3 Hz, 1H, ArH), 7.77-7.74 (m, 2H, ArH), 7.67-7.63 (m, 1H, ArH), 7.57-7.54 (m, 3H, ArH), 7.49 (d, J = 3.7 Hz, 1H, ArH), 6.64 (d, J = 3.5 Hz, 1H, ArH); ¹³C NMR (125 MHz, CDCl₃), δ: 168.4, 143.4, 138.8, 136.6, 133.9, 133.4, 132.9, 131.0, 129.6, 129.2, 115.9, 107.8. HRMS: (ESI, m/z); [M+H]⁺ calcd for 223.086 6; found 223.086 9.
Ⅰ-3	mp: 119-121 ℃。¹H NMR (500 MHz, CDCl₃), δ: 9.25 (s, 1H, ArH), 8.56 (d, J = 5.5 Hz, 1H, ArH), 7.61 (s, 1H, ArH), 7.55-7.53(m, 1H, ArH), 2.67-2.62 (m, 1H, CH), 1.48-1.44 (m, 2H, CH₂), 1.15-1.10 (m, 2H, CH₂); ¹³C NMR (125 MHz, CDCl₃), δ: 142.7, 135.7, 135.2, 131.5, 128.1, 114.2, 97.3, 32.0, 6.6. HRMS: (ESI, m/z); [M+H]⁺ calcd for 300.964 1; found 300.963 5.
Ⅱ-1	mp: 215-217 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.74 (s, 1H, ArH), 8.15-8.14 (m, 1H, ArH), 7.87 (d, J = 5.4 Hz, 1H, ArH), 7.79-7.78 (m, 1H, ArH), 7.50-7.46 (m, 2H, ArH), 7.34-7.31 (m, 1H, ArH), 7.11 (d, J = 7.5 Hz, 1H, ArH), 2.42 (s, 3H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆) δ: 138.5, 137.9, 135.0, 134.7, 129.0, 128.8, 127.2, 127.0, 126.4, 124.1, 115.2, 113.8, 113.4, 21.7. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 209.107 3; found: 209.108 4.
Ⅱ-2	mp: 214-217 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.12 (s, 1H, NH), 9.35 (d, J = 2.3 Hz, 1H, ArH), 8.86 (s, 1H, ArH), 8.67 (d, J = 2.6 Hz, 1H, ArH), 8.33 (d, J = 2.4 Hz, 1H, ArH), 8.27 (d, J = 5.5 Hz, 1H, ArH), 8.08 (t, J = 6.2 Hz, 2H, ArH), 8.02 (d, J = 8.2 Hz, 1H, ArH), 7.72-7.69 (m, 1H, ArH), 7.64-7.60 (m, 1H, ArH); ¹³C NMR (125 MHz, MeOD-d₄), δ: 150.6, 147.0, 139.1, 135.7, 135.5, 133.8, 131.8, 130.3, 130.0, 129.9, 129.7, 129.1, 128.9, 128.4, 115.3, 114.0. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 246.102 6; found: 246.102 2.
Ⅱ-3	mp: 211-213 ℃. ¹H NMR (500 MHz, MeOD-d₄) δ: 8.74 (s, 1H, ArH), 8.14 (d, J = 5.7 Hz, 1H, ArH), 7.89-7.87(m, 1H, ArH), 7.80 (s, 1H, ArH), 7.69-7.67 (m, 2H, ArH), 7.44 (t, J = 7.7 Hz, 2H, ArH), 7.31-7.26 (m, 1H, ArH). HRMS: (ESI, m/z); [M+H]⁺ calcd for: 195.091 7; found: 195.091 2.
Ⅱ-4	mp: 218-220 ℃. ¹H NMR (500 MHz, MeOD-d₄) δ: 8.71 (s, 1H, ArH), 8.08 (d, J = 5.6 Hz, 1H, ArH), 7.79 (s, 1H, ArH), 7.68 (d, J = 5.6 Hz, 1H, ArH), 7.55 (d, J = 9.3 Hz, 1H, ArH), 7.31-7.27 (m, 1H, ArH), 7.10 (d, J = 9.5 Hz, 1H, ArH), 7.06-7.02 (m, 1H, ArH), 3.85 (s, 3H, OCH₃); ¹³C NMR (125 MHz, DMSO-d₆) δ: 156.1, 137.7, 134.5, 133.3, 130.3, 129.5, 129.2, 127.4, 123.0, 120.6, 114.5, 111.6, 111.5, 55.3. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 225.102 2; found: 225.101 7.
Ⅲ-1	mp: 214-216 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.36 (s, 1H, NH), 8.83 (d, J = 1.1 Hz, 1H, ArH), 8.28 (d, J = 5.5 Hz, 1H, ArH), 7.94 (d, J = 5.5 Hz, 1H, ArH), 7.88 (s, 1H, ArH), 7.75 (s, 1H, ArH), 2.48 (s, 3H, CH₃), 2.32 (s, 3H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 206.7, 197.7, 140.0, 137.9, 135.80, 133.8, 132.5, 132.4, 132.0, 113.9, 109.0, 31.4, 26.5. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 229.097 2; found: 229.097 1.
Ⅲ-2	mp: 165-168 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.27 (s, 1H, NH), 8.82 (d, J = 1.1 Hz, 1H, ArH), 8.26 (d, J = 5.5 Hz, 1H, ArH), 7.92 (d, J = 5.3 Hz, 2H, ArH), 7.60 (s, 1H, ArH), 2.92 (q, J = 7.2 Hz, 2H, CH₂), 2.58 (q, J = 7.2 Hz, 2H, CH₂), 1.06-1.02 (m, 6H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 209.2, 199.6, 139.4, 136.9, 135.3, 133.3, 131.8, 131.3, 130.3, 113.4, 108.6, 36.0, 30.2, 8.4, 7.7. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 257.128 5; found: 257.129 1.
Ⅲ-3	mp: 180-181 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.41 (s, 1H, NH), 8.83 (d, J = 1.1 Hz, 1H, ArH), 8.25 (d, J = 5.5 Hz, 1H, ArH), 7.99 (s, 1H, ArH), 7.94 (s, 1H, ArH), 7.72-7.71 (m, 1H, ArH), 3.84 (s, 3H, OCH₃), 3.77 (s, 3H, OCH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 167.6, 164.9, 140.0, 135.6, 134.5, 133.6, 133.1, 131.6, 118.9, 113.2, 108.4, 52.8, 52.4. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 261.087 0; found: 261.087 0.
Ⅲ-4	mp: 171-173 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.75 (s, 1H, ArH), 8.23 (d, J = 5.7 Hz, 1H, ArH), 8.04–7.99 (m, 2H, ArH), 7.77-7.76 (m, 1H, ArH), 4.38 (q, J = 7.1 Hz, 2H, OCH₂), 4.31 (q, J = 7.1 Hz, 2H, OCH₂), 1.36-1.32 (m, 6H, CH₃); ¹³C NMR (125 MHz, MeOD-d₄), δ: 169.1, 166.7, 140.0, 135.7, 135.0, 134.9, 134.5, 134.2, 122.2, 114.7, 110.6, 62.9, 62.6, 14.7, 14.5. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 289.118 3; found: 289.118 6.
Ⅲ-5	mp: 163-164 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.38 (s, 1H, NH), 8.83 (d, J = 1.1 Hz, 1H, ArH), 8.25 (d, J = 5.6 Hz, 1H, ArH), 7.95 (d, J = 1.1 Hz, 2H, ArH), 7.72-7.71 (m, 1H, ArH), 4.24 (t, J = 6.5 Hz, 2H, OCH₂), 4.14 (t, J = 6.5 Hz, 2H, OCH₂), 1.67-1.62 (m, 4H, CH₂), 0.92 (t, J = 7.4 Hz, 3H, CH₃), 0.87 (t, J = 7.4 Hz, 3H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 167.1, 164.3, 139.7, 135.4, 133.7, 133.4, 132.7, 131.4, 119.6, 113.0, 108.3, 66.7, 66.2, 21.6, 21.3, 10.2. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 317.149 6; found: 317.150 4.
Ⅳ-1	mp: 129-135 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.64 (d, J = 1.1 Hz, 1H, ArH), 8.04 (d, J = 5.6 Hz, 1H, ArH), 7.66-7.65 (m, 1H, ArH), 7.50 (s, 1H, ArH), 7.11-7.07 (m, 2H, ArH), 6.73-6.71 (m, 2H, ArH), 6.63-6.60 (m, 1H, ArH), 4.45 (s, 2H, CH₂); ¹³C NMR (125 MHz, MeOD-d₄), δ: 150.4, 137.7, 135.4, 134.7, 133.5, 130.1, 129.7, 118.4, 115.3, 115.0, 114.5, 40.4. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 224.118 2; found: 224.119 0.
Ⅳ-2	mp: 155-156 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.64 (s, 1H, ArH), 8.04 (d, J = 5.6 Hz, 1H, ArH), 7.65-7.63 (m, 1H, ArH), 7.49 (s 1H, ArH), 7.05-7.02 (m, 2H, ArH), 6.66-6.63 (m, 2H, ArH), 4.42 (s, 2H, CH₂); ¹³C NMR (125 MHz, MeOD-d₄), δ: 149.0, 137.6, 135.3, 134.6, 133.3, 129.7, 129.6, 122.3, 117.5, 115.2, 114.5, 40.2. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 258.079 3; found: 258.080 1.
Ⅳ-3	mp: 156-160 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.60 (s, 1H, ArH), 8.00 (d, J = 5.6 Hz, 1H), 7.61-7.59 (m, 1H, ArH), 7.45 (s, 1H, ArH), 7.14-7.11 (m, 2H, ArH), 6.58–6.55 (m, 2H, ArH), 4.38 (s, 2H, CH₂); ¹³C NMR (125 MHz, MeOD-d₄), δ: 149.4, 137.6, 135.3, 134.7, 133.3, 132.6, 129.6, 115.7, 115.1, 114.4, 109.1, 40.1. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 302.028 7; found: 302.029 0.
Ⅳ-4	mp: 143-145 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 8.72 (s, 1H, ArH), 8.07 (d, J = 5.5 Hz, 1H, ArH), 7.63 (d, J = 5.5 Hz, 1H, ArH), 7.54 (s, 1H, ArH), 6.85 (d, J = 8.0 Hz, 2H, ArH), 6.58 (d, J = 8.1 Hz, 2H, ArH), 4.34 (s, 2H, CH₂), 2.11 (s, 3H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 146.7, 137.3, 134.4, 133.6, 131.1, 129.4, 127.8, 124.4, 113.9, 113.2, 112.8, 38.7, 20.2. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 238.133 9; found: 238.134 7.
Ⅳ-5	mp: 146-148 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 8.71 (s, 1H, ArH), 8.07 (d, J = 5.5 Hz, 1H, ArH), 7.63 (d, J = 5.5 Hz, 1H, ArH), 7.54 (s, 1H, ArH), 6.68 (d, J = 8.9 Hz, 2H, ArH), 6.62 (d, J = 8.7 Hz, 2H, ArH), 4.32 (s, 2H, CH₂), 3.63 (s, 3H, OCH₃); ¹³C NMR (125 MHz, MeOD-d₄), δ: 153.9, 144.2, 137.5, 135.2, 134.5, 133.4, 129.8, 116.3, 115.7, 115.2, 114.9, 56.2, 41.5. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 254.128 8; found: 254.129 6.
Ⅳ-6	mp: 134-136 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 8.71 (s, 1H), 8.07 (d, J = 5.5 Hz, 1H, ArH), 7.63 (d, J = 5.5 Hz, 1H, ArH), 7.55 (s, 1H, ArH), 6.87 (t, J = 8.7 Hz, 2H, ArH), 6.65-6.63 (m, 2H, ArH), 4.34 (s, 2H, CH₂); ¹³C NMR (125 MHz, DMSO-d₆), δ: 154.5 (d, J = 230.8 Hz), 145.7, 137.4, 134.5, 133.6, 131.1, 127.9, 115.3 (d, J = 21.8 Hz), 113.9, 113.3 (d, J = 7.2 Hz), 112.8, 38.9. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 242.108 8; found: 242.109 1.

Compd.

mp, ¹H NMR, ¹³C NMR, HRMS-ESI

Ⅰ-1

Ⅰ-2

Ⅰ-3

Ⅱ-1

Ⅱ-2

Ⅱ-3

Ⅱ-4

Ⅲ-1

Ⅲ-2

Ⅲ-3

Ⅲ-4

Ⅲ-5

Ⅳ-1

Ⅳ-2

Ⅳ-3

Ⅳ-4

Ⅳ-5

Ⅳ-6

6-氮杂吲哚类化合物的合成及抗胰腺癌活性评价

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曹阳 ¹^,² , 李前 ² , 王亚玲 ² , 崔文慧 ² , 钱晨亮 ² , 司鑫鑫 ²^,^*

药学学报 | 研究论文 2024,59(10): 2828-2835

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药学学报 | 研究论文 2024, 59(10): 2828-2835

6-氮杂吲哚类化合物的合成及抗胰腺癌活性评价

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曹阳¹^,², 李前², 王亚玲², 崔文慧², 钱晨亮², 司鑫鑫²^,^*

作者信息

1.南京大学化学化工学院, 江苏南京 210093

2.江苏海洋大学, 江苏省海洋药物活性分子筛选重点实验室, 江苏连云港 222005

通讯作者:

^*司鑫鑫, E-mail: sixx@jou.edu.cn

Synthesis and activity evaluation of 6-azazindole derivatives for pancreatic cancer therapy

Yang CAO¹^,², Qian LI², Ya-ling WANG², Wen-hui CUI², Chen-liang QIAN², Xin-xin SI²^,^*

Affiliations

1. School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China

2. Jiangsu Key Laboratory of Marine Pharmaceutical Compound Screening, Jiangsu Ocean University, Lianyungang 222005, China

出版时间: 2024-10-12 doi: 10.16438/j.0513-4870.2024-0135

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摘要

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通过筛选内部化合物库, 鉴定出了具有一定抗胰腺癌活性的片段。经系统改造合成了4类共计18个化合物, 并进行了抗胰腺癌活性评价。化合物Ⅱ-1 (IC₅₀ = 6.40 ± 0.34 μmol·L^-1) 和Ⅱ-2 (IC₅₀ = 7.15 ± 0.51 μmol·L^-1) 活性表现突出。随后用细胞划痕实验及侵袭实验评价了Ⅱ-1的抗迁移能力及侵袭能力, 结果显示, Ⅱ-1具有良好的抗迁移能力及突出的抗侵袭能力。利用分子对接技术及分子动力学模拟技术, 锁定了Ⅱ-1的靶点为双特异性酪氨酸磷酸化调控激酶1A (DYRK1A)。经酶活测试Ⅱ-1和Ⅱ-2分别有48%及32%酶抑制能力。

关键词

6-氮杂吲哚 / 抗胰腺癌 / 双特异性酪氨酸磷酸化调控激酶1A / 合成 / 抗增殖

Abstract

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Fragment with some anti-pancreatic cancer activity was identified by screening our internal chemical library. Eighteen compounds in 4 classes were synthesized by systematic modification and their anti-pancreatic cancer activity were evaluated. Ⅱ-1 (IC₅₀ = 6.40 ± 0.34 μmol·L^-1) and Ⅱ-2 (IC₅₀ = 7.15 ± 0.51 μmol·L^-1) exhibited outstanding activity. Subsequently, the anti-migration ability and invasion ability of Ⅱ-1 was evaluated by wound healing assay and invasion assay, Ⅱ-1 exhibited good anti-migration ability and outstanding anti-invasion ability. Using molecular docking technology and molecular dynamics simulation technology, the potential target was locked on bispecific tyrosine phosphorylation regulates kinase 1A (DYRK1A). By enzyme activity testing, the inhibitory capacity of Ⅱ-1 and Ⅱ-2 was 48% and 32%, respectively.

Key words

6-azaindole / anti-pancreatic cancer / bispecific tyrosine phosphorylation regulates kinase 1A / synthesis / anti-proliferation

引用本文

曹阳, 李前, 王亚玲, 崔文慧, 钱晨亮, 司鑫鑫. 6-氮杂吲哚类化合物的合成及抗胰腺癌活性评价. 药学学报, 2024 , 59 (10) : 2828 -2835 . DOI: 10.16438/j.0513-4870.2024-0135

Yang CAO, Qian LI, Ya-ling WANG, Wen-hui CUI, Chen-liang QIAN, Xin-xin SI. Synthesis and activity evaluation of 6-azazindole derivatives for pancreatic cancer therapy[J]. Acta Pharmaceutica Sinica, 2024 , 59 (10) : 2828 -2835 . DOI: 10.16438/j.0513-4870.2024-0135

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胰腺癌是最具侵袭性和致命性的癌症之一, 死亡率与发病率相近, 病死率极高^[1]。常见的治疗手段包括手术治疗、化疗、免疫疗法、放疗等^[2]。化疗是治疗难以切除和转移性胰腺癌的主要方法, 常用的药物包括氟尿嘧啶类、吉西他滨、金属铂类、伊立替康类、紫杉醇等^[3]。然而, 这些药物在提高总生存率方面发挥的作用依然有限。因此, 广泛开发具有抗胰腺的药物, 特别是能提高生存率的药物非常关键。

常见的药物发现手段包括小分子化合物库和高通量筛选、计算机辅助药物设计(CADD)、天然产物、基因工程、已上市药物开发新疗效等^[4]手段。利用小分子化合物库和高通量技术发现先导化合物并进行深入研究, 是制药工业开发药物的主要方法之一^[5]。利用该方法, 筛选了课题组化合物库, 发现了一种具有一定活性的化合物片段, 即A-1, 化学名6-氮杂吲哚(图 1)。

该骨架类化合物在抗胰腺癌方面未见报道, 作为一种具有较大潜力的结构母核, 有必要探究其在抗胰腺癌活性方面的表现。希望通过在母核上多个位点修饰发现具有良好活性的抗胰腺癌化合物。

结果与讨论

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1 6-氮杂吲哚母核抗胰腺癌活性的测试

采用CCK-8法(细胞增殖毒性检测) 对所筛选出的活性片段A-1 (图 1) 进行抗胰腺癌活性测试, 进行了三组平行实验, 结果显示(图 2) 该母核结构能够抑制胰腺癌细胞(PANC-1) 的存活, 具有一定的抗胰腺癌活性, 接下来将对该母核片段进行结构修饰, 希望发现更多且抗胰腺癌活性更强的化合物。

2 化合物的合成

所有化合物通过合成路线1所示获得, 其结构经 ¹H NMR、¹³C NMR、HR-MS确认为目标结构。理化性质及谱图数据见表 1。

为了研究的全面性, 对具有该母核的简单取代衍生物也进行了抗胰腺癌活性研究(图 3)。

3 体外抗胰腺癌活性评价

对所有化合物的抗胰腺癌活性进行了评价, 结果显示, Ⅱ-1和Ⅱ-2在10 μmol·L^-1浓度下对胰腺癌细胞展现出显著的抑制活性。随后对2个化合物的IC₅₀ (24 h) 进行了研究, Ⅱ-1和Ⅱ-2的IC₅₀分别为6.40 ± 0.34和7.15 ± 0.51 μmol·L^-1 (图 4)。

4 构效关系分析

Ⅰ类化合物抗胰腺癌活性较差, 推测6-氮杂吲哚母核氮上活泼氢对活性影响较大。Ⅱ类化合物活性差异性大, 芳环或芳杂环及其环上的取代基及位置十分重要, 对比4个化合物可以得出芳环的3位取代基可能是提高活性关键。Ⅲ类化合物虽然有富含氧元素的支链, 但活性未见明显提高。Ⅳ类化合物引入的亚甲胺基未能提升活性。

5 抗迁移和侵袭能力评价

癌细胞的迁移和侵袭与肿瘤的发展密切相关^{[6, 7]}。利用划痕实验与侵袭实验评估了Ⅱ-1在抵抗癌细胞迁移和侵袭方面的能力。图 5展示了Ⅱ-1不同时间(24和48 h) 不同浓度(1、5、10 μmol·L^-1) 抑制癌细胞迁移情况, 结果显示, Ⅱ-1展现出良好的抗迁移能力。图 6展示了Ⅱ-1在48 h不同浓度下(1、5、10 μmol·L^-1) 的抗侵袭情况, 结果显示, Ⅱ-1展现出显著的抗侵袭能力, 较低浓度(1 μmol·L^-1) 下也表现出强的抗侵袭能力。

6 分子对接

II-1和II-2结构上与Liu等^[8]报道的DYRK1A抑制剂具有一定相似性。利用分子对接技术将Ⅱ-1、Ⅱ-2与DYRK1A (PDB代码：7aj2) 蛋白对接以评估化合物与蛋白的亲和力, 去氢骆驼蓬碱(harmine, 是一种具有抗癌和抗炎活性的天然DYRK抑制剂) 为阳性对照。Ⅱ-1、Ⅱ-2和去氢骆驼蓬碱与DYRK1A的亲和力分别为-8.976 7、-9.386 6、-8.920 3, Ⅱ-1和Ⅱ-2与DYRK1A蛋白之间表现出良好的结合力, 甚至优于去氢骆驼蓬碱。上述结果表明, Ⅱ-1和Ⅱ-2与DYRK1A之间有相互作用。

7 分子动力学模拟

为了进一步阐明DYRK1A与化合物Ⅱ-1之间的相互作用, 利用GROMACS对DYRK1A与Ⅱ-1进行了100 ns的分子动力学模拟(MD)。图 7A模拟了DYRK1A蛋白C-α原子与Ⅱ-1重原子在100 ns内的RMSD (均方根误差) 变化, DYRK1A~Ⅱ-1配合物在约90 ns后趋于稳定。图 7B展示了Ⅱ-1与DYRK1A的结合模式。Ⅱ-1的核心骨架6-氮杂吲哚环与Lys188有氢键相互作用, 与Val173、Ile165、Ala186、Met240、Phe238、Leu241、Leu294和Val306有疏水键作用。利用分子力学泊松-玻尔兹曼表面积法(MM-PBSA) 基于最后5 ns模拟的结合构象计算平均结合能, 化合物Ⅱ-1的结合自由能为-137.08 ± 11.12 kJ·mol^-1。进一步验证了Ⅱ-1与DYRK1A有强相互作用, 推测DYRK1A可能是靶点之一。

8 DYRK1A活性评价

对Ⅱ-1和Ⅱ-2进行了DYRK1A酶活测试。Ⅱ-1能将DYRK1A酶活力降低48%, Ⅱ-2能降低35%, 说明DYRK1A是Ⅱ-1和Ⅱ-2的靶点之一。

9 小结

通过筛选内部化合物库的方法鉴定出了具有一定抗胰腺癌活性的6-氮杂吲哚母核, 合成了一系列6-氮杂吲哚衍生物, 并进行了抗胰腺癌活性评价。Ⅱ-1和Ⅱ-2表现出优异的抗胰腺癌活性。对Ⅱ-1的抗迁移能力及侵袭能力进行了深入研究, 发现具有较好的抵抗作用。利用分子对接、分子动力学将靶点锁定为DYRK1A, 酶活测试证实化合物Ⅱ-1和Ⅱ-2具有酶抑制作用。

实验部分

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核磁共振波谱仪, Avance Ⅲ 500 MHZ型, 内标为TMS; 液质联用仪, Q Exactive HF型; 薄层色谱硅胶, 山西诺泰生物; 柱色谱硅胶(200~300目), 烟台新诺; 中性氧化铝(100~200目), 上海盛亚; 实验所用试剂均为化学纯或分析纯, 购自毕得医药与上海泰坦公司。

化学实验

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1.1 合成方法

1.1.1 Ⅰ类化合物的合成步骤

以(3-溴-1H-吡咯并[2, 3-c]吡啶)-1-环丁甲酰胺(Ⅰ-1) 为例。将化合物3-溴-6-氮杂吲哚100 mg (0.51 mmol) 溶解在4 mL二氯甲烷(DCM) 中, 加入4-二甲氨基吡啶(DMAP) 6 mg (0.05 mmol), 环丁甲酰氯88 μL (0.77 mmol) 用1 mL DCM稀释室温搅拌下缓慢加入反应体系(持续5 min), 加三乙胺212 μL (1.53 mmol)。室温下反应12 h, TLC检测反应完成^{[9, 10]}。反应液用饱和NH₄Cl水溶液淬灭, DCM (10 mL×3) 萃取3次, 合并有机相, 无水Na₂SO₄干燥, 旋干溶剂。用硅胶柱色谱纯化得Ⅰ-1 (乙酸乙酯-石油醚, 1∶5)。

1.1.2 Ⅱ类化合物的合成步骤

以3-苯基(3-甲基)-1H-吡咯并[2, 3-c]吡啶(Ⅱ-1) 为例。取化合物3-溴-6-氮杂吲哚100 mg (0.51 mmol), 依次加入XPhos Pd G2 40 mg (0.05 mmol), XPhos 24 mg (0.05 mmol), 无水K₃PO₄ 324 mg (1.53 mmol), 水0.76 mL, 1, 4二氧六环2.4 mL, N₂置换3次, 100 ℃下反应4 h^[11]。旋干反应液, 乙酸乙酯(10 mL×3) 萃取3次, 合并有机相, 无水Na₂SO₄干燥, 旋干溶剂。用硅胶柱色谱纯化得到Ⅱ-1 (甲醇-二氯甲烷, 1∶20)。

1.1.3 Ⅲ类化合物的合成步骤

以3-((1H-吡咯并[2, 3-c]吡啶)-3)-烯-2, 4-戊二酮(Ⅲ-1) 为例。取化合物6-氮杂吲哚-3-羧醛100 mg (0.68 mmol), 5 mL甲醇(MeOH) 溶解, 依次加入乙酰丙酮105 μL (1.03 mmol), 哌啶7 μL (0.07 mmol), 60 ℃反应3 h^{[12, 13]}。TLC检测反应结束, 旋干反应液, 硅胶柱色谱纯化得Ⅲ-1 (甲醇-二氯甲烷, 1∶20)。

1.1.4 Ⅳ类化合物的合成步骤

以N-((1H-吡咯并[2, 3-c]吡啶)-3)-亚甲基苯胺(Ⅳ-1) 为例。取化合物6-氮杂吲哚-3-羧醛100 mg (0.68 mmol), 5 mL乙醇溶解, 加苯胺94 μL (1.03 mmol), 50 ℃下反应3 h。TLC检测反应结束, 加NaBH₄ 260 mg (6.8 mmol) 室温反应2 h, TLC检测反应结束^{[14, 15]}。加水至不再有气泡产生为止, 乙酸乙酯(10 mL×3) 萃取3次, 合并有机相, 无水Na₂SO₄干燥, 旋干溶剂, 用中性氧化铝柱色谱纯化得到Ⅳ-1 (甲醇-二氯甲烷, 1∶20)。

1.2 胰腺癌细胞存活率测试

采用CCK-8法测定化合物处理后的细胞存活率。在37 ℃含5% CO₂的环境下用含10%胎牛血清的DMEM培养基(改良杜氏伊格尔培养基) 培养胰腺癌细胞(PANC-1)。收集对数期的细胞, 以5×10³个/孔的密度接种于96孔板中, 继续培养24 h。更换为含10 μmol·L^-1化合物的培养基或含1‰对照溶剂的培养液(DMSO与培养基体积比1∶1 000) 继续培养48 h。在每个孔中加入10 µL CCK-8溶液孵育2 h。用CCK-8法测定化合物处理后的细胞存活率。根据吸光度计算细胞抑制率, 存活率= (给药组-空白组) / (阴性对照组-空白组) × 100%, 抑制率= 100% - 存活率。显著性差异用P值来表示, P值利用GraphPad Prism 9制作及计算。

1.3 IC₅₀值测试

在37 ℃含5% CO₂的环境下用含10%胎牛血清的DMEM培养基培养PANC-1细胞。收集对数期的细胞, 以5×10³个/孔的密度接种于96孔板中, 继续培养24 h。更换为含1‰对照溶剂的培养液(DMSO与培养基体积比1∶1 000) 或含不同浓度(100、10、1、0.1、0.01 μmol·L^-1) 化合物的培养液继续培养48 h。然后在每个孔中加入10 µL CCK-8溶液孵育2 h。最后, 在酶标仪上450 nm波长下测定吸光度。根据吸光度计算细胞抑制率, 存活率 = (给药组-空白组) / (阴性对照组-空白组) × 100%, 抑制率= 100% - 存活率。使用GraphPad Prism 9软件基于抑制率计算IC₅₀值。

1.4 分子对接

人类DYRK1A蛋白(PDB代码: 7aj2) 的晶体结构从PDB数据库(https://www.rcsb.org/) 下载。DYRK1A蛋白由Discovery Studio (DS) 软件中的“Clean protein”模块进行准备。使用Smina进行分子对接, 以活性配体坐标为中心, 半径为8 Å为活性位点^[16]。利用Lamarckian Genetic算法(LGA) 实现蛋白与配体的对接。得分最高的位置是针对每个分子的。评分函数为“最小化亲和力”评分。

1.5 分子动力学模拟

使用Smina将Ⅱ-1与DYRK1A蛋白重新对接, 选择合适的构象作为动力学模拟复合物的初始结合构象。分子动力学模拟使用GROMACS (2019.4版本) 软件, 并对每个DYRK1A配体复合物进行分子动力学模拟。配体的电荷和力场参数由CHARMM-GUI (https://www.charmm-gui.org/) 生成, 配体和蛋白的力场分别为GAFF2和AMBER ff19SB^[17]。整个系统由27767水(TIP3P) 溶剂化^[18]。然后, 水相在每个维度上从系统外围延伸10 Å。之后, 向系统中分别加入78个K⁺和89个Cl^-离子使系统总电荷为零。MD模拟包括能量最小化、平衡和生产。模拟开始于基于最速下降算法的5 000步能量最小化。在平衡阶段, 包括NVT条件下的500 ps模拟和NPT条件下的500 ps模拟。使用Parrinello Rahman将系统保持在1 atm的压力下, 运用V-rescale将系统维持在300 K的恒定温度。最后, 在NPT下进行100 ns MD模拟, 每500 ps保存一次复合体坐标。

采用MM/PBSA法估算配体与蛋白质之间的结合能^[19]。从95~100 ns的MD模拟轨迹中每5 ps提取一次快照。以快照作为输入, 实现MM/PBSA方法的g_mmpbsa (http://rashmikumari.github.io/g_mmpbsa/) 计算每个蛋白质-配体复合物的结合自由能。平均结合能表示蛋白质与配体的结合亲和力。

1.6 DYRK1A酶活性测试

使用Promega公司的ADP-Glo^TM Max Assay试剂盒检测DYRK1A激酶活性。通过检测激酶反应中生成的ADP来反应激酶活性, 在ADP生成反应完成后, 加入等体积的ADP-Glo^TM试剂终止反应, 并消耗尽剩余的ATP; 加入ADP-Glo^TM Max检测试剂, 将ADP转变为ATP的同时, 在偶联的萤光素酶/荧光素反应中将ATP转变为光信号。根据试剂盒操作说明书, 向反应板中每孔加入2 μL DYRK1A激酶(0.2 ng·μL^-1), 加入1 μL待测化合物(终浓度10 μmol·L^-1) 彻底振荡混匀, 离心1 min, 共同在EP管中25 ℃孵育30 min。加入2 μL ATP (30 μmol·L^-1) 和DYRK1A底物(0.1 mg·mL^-1) 混合液后, 将反应混合物在25 ℃下孵育60 min。之后, 加入4 μL ADP-Glo激酶试剂以停止激酶反应, 并在25 ℃下孵育40 min。然后, 加入8 μL激酶检测试剂并在25 ℃下再孵育40 min。发光信号(相对发光单位) 记录在读板器(BMG Labtech, PHERAstar FSX) 上。

1.7 划痕实验

将胰腺癌细胞放在6孔板中, 培养至融合。使用无菌移液管尖端在细胞单层中创建一个划痕, 确保伤口的大小和宽度相似, 在形成伤口后, 用培养基轻轻清洗细胞, 以清除任何脱落的细胞或碎片。在各孔中加入化合物或DMSO, 再孵育48 h, 使用相衬显微镜定期间隔(0、24、48 h) 拍摄初始受伤区域的图像, 最后使用图像分析软件测量伤口闭合或迁移距离。

1.8 侵袭实验

上室预涂Matrigel (37 ℃, 30 min), 填充含5×10⁴个细胞的无血清DMEM, 下室填充含10%胎牛血清的DMEM, 37 ℃孵育48 h后, 收集下室细胞, 室温0.5%结晶紫(质量分数) 染色20 min。用倒置光学显微镜对染色细胞计数。

1.9 数据统计分析

实验数据采用GraphPad Prism 9软件进行统计分析, 所有实验数据以三个独立实验的平均值± SD表示, 两组间数据的比较采用单因素方差分析和Dunnett比较检验进行差异分析, 当P < 0.05时, 差异具有统计显著性。

作者贡献: 曹阳完成化合物合成路线设计、结构解析、论文稿件润色; 李前完成化合物合成、表征以及论文初稿撰写; 王亚玲完成分子对接及分子动力学模拟; 钱晨亮完成了细胞活性测试及酶活测试; 崔文慧完成审核论文; 司鑫鑫完成实验设计、提供学术指导、审核论文。

利益冲突: 所有作者声明不存在利益冲突。

基金

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江苏海洋大学研究生科研与实践创新计划项目(KYCX2022-31)
中国博士后科学基金(2023M741444)

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2024年第59卷第10期

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doi: 10.16438/j.0513-4870.2024-0135

接收时间：2024-02-14
首发时间：2025-11-24
出版时间：2024-10-12

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收稿日期：2024-02-14
修回日期：2024-07-19

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江苏海洋大学研究生科研与实践创新计划项目(KYCX2022-31)

中国博士后科学基金(2023M741444)

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1.南京大学化学化工学院, 江苏南京 210093

2.江苏海洋大学, 江苏省海洋药物活性分子筛选重点实验室, 江苏连云港 222005

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^*司鑫鑫, E-mail: sixx@jou.edu.cn

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2种不同金属材料的力学参数

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

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Compd.	mp, ¹H NMR, ¹³C NMR, HRMS-ESI
Ⅰ-1	mp: 159-162 ℃. ¹H NMR (500 MHz, CDCl₃), δ: 9.72 (s, 1H, ArH), 8.54 (d, J = 5.3 Hz, 1H, ArH), 7.50 (s, 1H, ArH), 7.46 (d, J = 5.3 Hz, 1H, ArH), 3.78 (p, J = 8.4 Hz, 1H, -CH), 2.61-2.53 (m, 2H, CH₂), 2.45-2.38 (m, 2H, CH₂), 2.19-2.00 (m, 2H, CH₂); ¹³C NMR (125 MHz, CDCl₃), δ: 172.0, 143.5, 138.7, 134.9, 132.0, 126.5, 113.9, 98.4, 39.0, 25.2, 18.3. HRMS: (ESI, m/z); [M+H]⁺ calcd for 279.012 8; found 279.013 8.
Ⅰ-2	mp: 102-105 ℃. ¹H NMR (500 MHz, CDCl₃), δ: 9.64 (s, 1H, ArH), 8.49 (d, J = 5.3 Hz, 1H, ArH), 7.77-7.74 (m, 2H, ArH), 7.67-7.63 (m, 1H, ArH), 7.57-7.54 (m, 3H, ArH), 7.49 (d, J = 3.7 Hz, 1H, ArH), 6.64 (d, J = 3.5 Hz, 1H, ArH); ¹³C NMR (125 MHz, CDCl₃), δ: 168.4, 143.4, 138.8, 136.6, 133.9, 133.4, 132.9, 131.0, 129.6, 129.2, 115.9, 107.8. HRMS: (ESI, m/z); [M+H]⁺ calcd for 223.086 6; found 223.086 9.
Ⅰ-3	mp: 119-121 ℃。¹H NMR (500 MHz, CDCl₃), δ: 9.25 (s, 1H, ArH), 8.56 (d, J = 5.5 Hz, 1H, ArH), 7.61 (s, 1H, ArH), 7.55-7.53(m, 1H, ArH), 2.67-2.62 (m, 1H, CH), 1.48-1.44 (m, 2H, CH₂), 1.15-1.10 (m, 2H, CH₂); ¹³C NMR (125 MHz, CDCl₃), δ: 142.7, 135.7, 135.2, 131.5, 128.1, 114.2, 97.3, 32.0, 6.6. HRMS: (ESI, m/z); [M+H]⁺ calcd for 300.964 1; found 300.963 5.
Ⅱ-1	mp: 215-217 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.74 (s, 1H, ArH), 8.15-8.14 (m, 1H, ArH), 7.87 (d, J = 5.4 Hz, 1H, ArH), 7.79-7.78 (m, 1H, ArH), 7.50-7.46 (m, 2H, ArH), 7.34-7.31 (m, 1H, ArH), 7.11 (d, J = 7.5 Hz, 1H, ArH), 2.42 (s, 3H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆) δ: 138.5, 137.9, 135.0, 134.7, 129.0, 128.8, 127.2, 127.0, 126.4, 124.1, 115.2, 113.8, 113.4, 21.7. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 209.107 3; found: 209.108 4.
Ⅱ-2	mp: 214-217 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.12 (s, 1H, NH), 9.35 (d, J = 2.3 Hz, 1H, ArH), 8.86 (s, 1H, ArH), 8.67 (d, J = 2.6 Hz, 1H, ArH), 8.33 (d, J = 2.4 Hz, 1H, ArH), 8.27 (d, J = 5.5 Hz, 1H, ArH), 8.08 (t, J = 6.2 Hz, 2H, ArH), 8.02 (d, J = 8.2 Hz, 1H, ArH), 7.72-7.69 (m, 1H, ArH), 7.64-7.60 (m, 1H, ArH); ¹³C NMR (125 MHz, MeOD-d₄), δ: 150.6, 147.0, 139.1, 135.7, 135.5, 133.8, 131.8, 130.3, 130.0, 129.9, 129.7, 129.1, 128.9, 128.4, 115.3, 114.0. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 246.102 6; found: 246.102 2.
Ⅱ-3	mp: 211-213 ℃. ¹H NMR (500 MHz, MeOD-d₄) δ: 8.74 (s, 1H, ArH), 8.14 (d, J = 5.7 Hz, 1H, ArH), 7.89-7.87(m, 1H, ArH), 7.80 (s, 1H, ArH), 7.69-7.67 (m, 2H, ArH), 7.44 (t, J = 7.7 Hz, 2H, ArH), 7.31-7.26 (m, 1H, ArH). HRMS: (ESI, m/z); [M+H]⁺ calcd for: 195.091 7; found: 195.091 2.
Ⅱ-4	mp: 218-220 ℃. ¹H NMR (500 MHz, MeOD-d₄) δ: 8.71 (s, 1H, ArH), 8.08 (d, J = 5.6 Hz, 1H, ArH), 7.79 (s, 1H, ArH), 7.68 (d, J = 5.6 Hz, 1H, ArH), 7.55 (d, J = 9.3 Hz, 1H, ArH), 7.31-7.27 (m, 1H, ArH), 7.10 (d, J = 9.5 Hz, 1H, ArH), 7.06-7.02 (m, 1H, ArH), 3.85 (s, 3H, OCH₃); ¹³C NMR (125 MHz, DMSO-d₆) δ: 156.1, 137.7, 134.5, 133.3, 130.3, 129.5, 129.2, 127.4, 123.0, 120.6, 114.5, 111.6, 111.5, 55.3. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 225.102 2; found: 225.101 7.
Ⅲ-1	mp: 214-216 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.36 (s, 1H, NH), 8.83 (d, J = 1.1 Hz, 1H, ArH), 8.28 (d, J = 5.5 Hz, 1H, ArH), 7.94 (d, J = 5.5 Hz, 1H, ArH), 7.88 (s, 1H, ArH), 7.75 (s, 1H, ArH), 2.48 (s, 3H, CH₃), 2.32 (s, 3H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 206.7, 197.7, 140.0, 137.9, 135.80, 133.8, 132.5, 132.4, 132.0, 113.9, 109.0, 31.4, 26.5. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 229.097 2; found: 229.097 1.
Ⅲ-2	mp: 165-168 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.27 (s, 1H, NH), 8.82 (d, J = 1.1 Hz, 1H, ArH), 8.26 (d, J = 5.5 Hz, 1H, ArH), 7.92 (d, J = 5.3 Hz, 2H, ArH), 7.60 (s, 1H, ArH), 2.92 (q, J = 7.2 Hz, 2H, CH₂), 2.58 (q, J = 7.2 Hz, 2H, CH₂), 1.06-1.02 (m, 6H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 209.2, 199.6, 139.4, 136.9, 135.3, 133.3, 131.8, 131.3, 130.3, 113.4, 108.6, 36.0, 30.2, 8.4, 7.7. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 257.128 5; found: 257.129 1.
Ⅲ-3	mp: 180-181 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.41 (s, 1H, NH), 8.83 (d, J = 1.1 Hz, 1H, ArH), 8.25 (d, J = 5.5 Hz, 1H, ArH), 7.99 (s, 1H, ArH), 7.94 (s, 1H, ArH), 7.72-7.71 (m, 1H, ArH), 3.84 (s, 3H, OCH₃), 3.77 (s, 3H, OCH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 167.6, 164.9, 140.0, 135.6, 134.5, 133.6, 133.1, 131.6, 118.9, 113.2, 108.4, 52.8, 52.4. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 261.087 0; found: 261.087 0.
Ⅲ-4	mp: 171-173 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.75 (s, 1H, ArH), 8.23 (d, J = 5.7 Hz, 1H, ArH), 8.04–7.99 (m, 2H, ArH), 7.77-7.76 (m, 1H, ArH), 4.38 (q, J = 7.1 Hz, 2H, OCH₂), 4.31 (q, J = 7.1 Hz, 2H, OCH₂), 1.36-1.32 (m, 6H, CH₃); ¹³C NMR (125 MHz, MeOD-d₄), δ: 169.1, 166.7, 140.0, 135.7, 135.0, 134.9, 134.5, 134.2, 122.2, 114.7, 110.6, 62.9, 62.6, 14.7, 14.5. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 289.118 3; found: 289.118 6.
Ⅲ-5	mp: 163-164 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 12.38 (s, 1H, NH), 8.83 (d, J = 1.1 Hz, 1H, ArH), 8.25 (d, J = 5.6 Hz, 1H, ArH), 7.95 (d, J = 1.1 Hz, 2H, ArH), 7.72-7.71 (m, 1H, ArH), 4.24 (t, J = 6.5 Hz, 2H, OCH₂), 4.14 (t, J = 6.5 Hz, 2H, OCH₂), 1.67-1.62 (m, 4H, CH₂), 0.92 (t, J = 7.4 Hz, 3H, CH₃), 0.87 (t, J = 7.4 Hz, 3H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 167.1, 164.3, 139.7, 135.4, 133.7, 133.4, 132.7, 131.4, 119.6, 113.0, 108.3, 66.7, 66.2, 21.6, 21.3, 10.2. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 317.149 6; found: 317.150 4.
Ⅳ-1	mp: 129-135 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.64 (d, J = 1.1 Hz, 1H, ArH), 8.04 (d, J = 5.6 Hz, 1H, ArH), 7.66-7.65 (m, 1H, ArH), 7.50 (s, 1H, ArH), 7.11-7.07 (m, 2H, ArH), 6.73-6.71 (m, 2H, ArH), 6.63-6.60 (m, 1H, ArH), 4.45 (s, 2H, CH₂); ¹³C NMR (125 MHz, MeOD-d₄), δ: 150.4, 137.7, 135.4, 134.7, 133.5, 130.1, 129.7, 118.4, 115.3, 115.0, 114.5, 40.4. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 224.118 2; found: 224.119 0.
Ⅳ-2	mp: 155-156 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.64 (s, 1H, ArH), 8.04 (d, J = 5.6 Hz, 1H, ArH), 7.65-7.63 (m, 1H, ArH), 7.49 (s 1H, ArH), 7.05-7.02 (m, 2H, ArH), 6.66-6.63 (m, 2H, ArH), 4.42 (s, 2H, CH₂); ¹³C NMR (125 MHz, MeOD-d₄), δ: 149.0, 137.6, 135.3, 134.6, 133.3, 129.7, 129.6, 122.3, 117.5, 115.2, 114.5, 40.2. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 258.079 3; found: 258.080 1.
Ⅳ-3	mp: 156-160 ℃. ¹H NMR (500 MHz, MeOD-d₄), δ: 8.60 (s, 1H, ArH), 8.00 (d, J = 5.6 Hz, 1H), 7.61-7.59 (m, 1H, ArH), 7.45 (s, 1H, ArH), 7.14-7.11 (m, 2H, ArH), 6.58–6.55 (m, 2H, ArH), 4.38 (s, 2H, CH₂); ¹³C NMR (125 MHz, MeOD-d₄), δ: 149.4, 137.6, 135.3, 134.7, 133.3, 132.6, 129.6, 115.7, 115.1, 114.4, 109.1, 40.1. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 302.028 7; found: 302.029 0.
Ⅳ-4	mp: 143-145 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 8.72 (s, 1H, ArH), 8.07 (d, J = 5.5 Hz, 1H, ArH), 7.63 (d, J = 5.5 Hz, 1H, ArH), 7.54 (s, 1H, ArH), 6.85 (d, J = 8.0 Hz, 2H, ArH), 6.58 (d, J = 8.1 Hz, 2H, ArH), 4.34 (s, 2H, CH₂), 2.11 (s, 3H, CH₃); ¹³C NMR (125 MHz, DMSO-d₆), δ: 146.7, 137.3, 134.4, 133.6, 131.1, 129.4, 127.8, 124.4, 113.9, 113.2, 112.8, 38.7, 20.2. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 238.133 9; found: 238.134 7.
Ⅳ-5	mp: 146-148 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 8.71 (s, 1H, ArH), 8.07 (d, J = 5.5 Hz, 1H, ArH), 7.63 (d, J = 5.5 Hz, 1H, ArH), 7.54 (s, 1H, ArH), 6.68 (d, J = 8.9 Hz, 2H, ArH), 6.62 (d, J = 8.7 Hz, 2H, ArH), 4.32 (s, 2H, CH₂), 3.63 (s, 3H, OCH₃); ¹³C NMR (125 MHz, MeOD-d₄), δ: 153.9, 144.2, 137.5, 135.2, 134.5, 133.4, 129.8, 116.3, 115.7, 115.2, 114.9, 56.2, 41.5. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 254.128 8; found: 254.129 6.
Ⅳ-6	mp: 134-136 ℃. ¹H NMR (500 MHz, DMSO-d₆), δ: 8.71 (s, 1H), 8.07 (d, J = 5.5 Hz, 1H, ArH), 7.63 (d, J = 5.5 Hz, 1H, ArH), 7.55 (s, 1H, ArH), 6.87 (t, J = 8.7 Hz, 2H, ArH), 6.65-6.63 (m, 2H, ArH), 4.34 (s, 2H, CH₂); ¹³C NMR (125 MHz, DMSO-d₆), δ: 154.5 (d, J = 230.8 Hz), 145.7, 137.4, 134.5, 133.6, 131.1, 127.9, 115.3 (d, J = 21.8 Hz), 113.9, 113.3 (d, J = 7.2 Hz), 112.8, 38.9. HRMS: (ESI, m/z); [M+H]⁺ calcd for: 242.108 8; found: 242.109 1.

Compd.

mp, ¹H NMR, ¹³C NMR, HRMS-ESI

Ⅰ-1

Ⅰ-2

Ⅰ-3

Ⅱ-1

Ⅱ-2

Ⅱ-3

Ⅱ-4

Ⅲ-1

Ⅲ-2

Ⅲ-3

Ⅲ-4

Ⅲ-5

Ⅳ-1

Ⅳ-2

Ⅳ-3

Ⅳ-4

Ⅳ-5

Ⅳ-6