General
Melting points were determined on a Gallan-Kamp apparatus and are uncorrected. The IR spectra were recorded on a Shimadzu 470 IR-spectrophotometer (KBr; νmax in cm−1). The 1H and 13C NMR spectra were recorded on a Varian A5 500 MHz spectrometer using DMSO-d6 (except for compounds 3 and 5a in CDCl3) as a solvent and tetramethylsilane (TMS) as internal reference. Coupling constants (J values) are given in Hertz (Hz). The purity of the obtained products is checked by TLC.
Reaction of 2-acetylcyclohexanones 1a,b with cyanothioacetamide; Synthesis compounds 2a,b; general method.
A mixture of compound 1a,b (10 mmol), cyanothioacetamide (10 mmol) and piperidine (0..8 mL, 10 mmol) in ethanol (100 mL) was refluxed for 2 h. The yellow crystals that formed on hot were collected, washed with methanol, dried in air to give compounds 2a,b. The purity of these products is 100% and needs no any purification.
7-Acetyl-4-cyano-1,6-dimethyl-6-hydroxy-8-(3-nitrophenyl)-5,6,7,8-tetra-hydroisoquinoline-3(2 H )-thione (2a). It was synthesized by reaction of 1a with cyanothioacetamide. Yield: 96%; m. p: 279-280°C. IR: 3429 (O-H), 3235 (N-H); 3139 (C-H, sp2); 2971 (C-H, sp3); 2221 (C≡N); 1710 (C=O). 1H NMR: δ 13.68 (s, 1H, NH); 7.95-8.05 (m, 2H, ArH); 7.51-7.58 (m, 2H, ArH); 5.05 (s, 1H, OH); 4.61-4.63 (d, J =10, 1H, C8H); 3.23-3.26 (d, J =15, 1H, C5H), 2.88-2.90 (d, J =10, 1H, C7H), 2.83-2.87 (d, J =20, 1H, C5H); 2.12 (s, 3H, COCH3); 1.86 (s, 3H, CH3);1.23 (s, 3H, CH3). Anal. calcd for C20H19N3O4S (397.11): C, 60.44; H, 4.82; N, 10.57%. Found: C, 60.67; H, 5.11; N, 10.28%.
7-Acetyl-4-cyano-1,6-dimethyl-6-hydroxy-8-(4-nitrophenyl)-5,6,7,8-tetra-hydroisoquinoline-3(2 H )-thione (2b). It was synthesized by reaction of 1b with cyanothioacetamide. Yield: 93%; m. p 290-291°C. IR: 3482 (O-H); 3235 (NH); 3106 (C-H, sp2); 2971, 2872 (C-H, sp3); 2220 (C≡N); 1708 (C=O). 1H NMR: δ13.83 (s, H, NH), 7.84-7.86 (d, J =10, H, ArH); 7.62-7.64 (d, J =10, H, ArH); 7.51-7.53 (d, J =10, H, ArH); 7.33-7.34 (d, J =5, H, Ar), 5.04 (s, 1H, OH); 4.97-4.99 (d, J =10, 1H, C8H); 3.33-3.16 (d, J =20, 1H, C5H), 3.16-3.10 (d, J =5, 1H, C7H), 2.86-2.90 (d, J =20, 1H, C5H); 2.02 (s, 3H, COCH3); 1.93 (s, 3H, CH3); 1.29 (s, 3H, CH3). Anal. calcd for C20H19N3O4S (397.11): C, 60.44; H, 4.82; N, 10.57%. Found: C, 60.32; H, 5.04; N, 10.33%.
Reaction of compounds 2a,b with ethyl iodide, 2-chloroacetamide (4a) or its N-aryl-2-chloroacetamides 4b-f; Synthesis of compounds 3 and 5a-j; general method.
A mixture of 2a,b (10 mmol), ethyl iodide, 2-chloroacetamide (4a) or N-aryl-2-chloroacetamides 4b-f (10 mmol) and sodium acetate trihydrate (1.50 g, 11 mmol) in ethanol (100 mL) was refluxed for one hour. The solid that formed after cooling (50 mL) was collected and then recrystallized from ethanol to give white crystals of compounds 3 and 5a-i.
7-Acetyl-4-cyano-1,6-dimethyl-3-ethylthio-6-hydroxy-8-(4-nitrophenyl)-5,6,7,8-tetrahydroisoquinoline (3): It was synthesized by reaction of 2b with ethyl iodide. Yield: 83%; m.p.: 144-145°C. IR: 3509 (O-H); 3098 (C-H, sp2); 2974, 2919 (C-H, sp3); 2213 (C≡N); 1698 (C=O), 1603(C=N). 1H NMR: δ 8.13-8.15 (d, J = 10, 2H, ArH), 7.35-7.37 (d, J = 10, 2H, ArH), 4.99 (s, 1H, OH), 4.75-4.78 (d, J = 15, 1H, C8H), 3.15-3.31 (m, 3H: C5H and SCH2), 2.87-2.95 (m, 2H: C7H and C5H), 2.18 (s, 3H, COCH3), 1.98 (s, 3H, CH3), 1.31 (s, 3H, CH3), 1.29 (t, 3H, CH3). Anal. Calcd for C22H23N3O4S ( 425.14): C, 62.10; H, 5.45; N, 9.88%. Found: C, 62.37; H, 5.18; N, 10.01%.
2-[(7-Acetyl-4-cyano-1,6-dimethyl-6-hydroxy-8-(3-nitrophenyl)-5,6,7,8-tetrahydro-isoquinolin-3-yl)thio]acetamide (5a): It was synthesized by reaction of 2a with 2-chloroacetamide (4a). Yield: 91%; m.p.: 174-175°C. IR: 3481, 3373 (O-H, NH2); 2991, 2930 (C-H, sp3); 2215 (C≡N); 1701 (C=O, acetyl); 1660 (C=O, amide). 1H NMR: δ 7.26-8.18 (m, 6H: NH2 and ArH), 4.53-4.55 (d, J = 10.0, 1H, OH), 3.82- 3.97 (dd, J = 15.0, 2H: C7H and C8H), 3.02-3.21(m, 4H: SCH2 and C5H2), 1.96 (s, 3H, COCH3), 1.87 (s, 3H, CH3), 1.42 (s, 3H, CH3). 13C NMR: δ 214.79, 175.43, 161.92, 160.77, 160.44, 160.11, 159.78, 158.14, 149.88, 149.66, 145.64, 134.84, 131.34, 129.23, 123.41, 122.78, 118.74, 116.47, 114.65, 114.20, 111.93, 106.45, 69.90, 64.12, 45.89, 42.55, 35.59, 33.60, 28.30, 25.83. Anal. Calcd. for C22H22N4O5S (454.13): C, 58.14; H, 4.88; N, 12.33%. Found: C, 58.00; H, 5.03; N, 11.98%.
2-[(7-Acetyl-4-cyano-1,6-dimethyl-6-hydroxy-8-(3-nitrophenyl)-5,6,7,8-tetrahydro-isoquinolin-3-yl)thio]- N -phenylacetamide (5b). It was synthesized by reaction of 2a with N-phenyl-2-chloroacetamide (4b).Yield: 93%; m.p.: 191-192°C. IR: 3467 (O-H); 3335(N-H); 3063 (C-H, sp2); 2999, 2914 (C-H, sp3); 2214 (C≡N); 1702 (C=O, acetyl); 1687 (C=O, amide). 1H NMR: δ 10.25 (s, 1H, NH), 8.06-8.08 (d, 1H, ArH), 7.94-7.95 (d, 1H, ArH), 7.51-7.56 (m, 4H, ArH), 7.24-7.28 (m, 2H, ArH), 7.00-7.04 (m, 1H, ArH), 5.00 (s, 1H, OH), 4.76-4.79 (d, J = 15, 1H, C8H), 4.08-4.18 (dd, J = 15, 2H, SCH2), 3.45 (m, 1H, C5H), 2.93-2.97 (m, 2H: C7H and C5H), 2.19 (s, 3H, COCH3), 1.91 (s, 3H, CH3), 1.28 (s, 3H, CH3). Anal. Calcd. for C28H26N4O5S (530.16): C, 63.38; H, 4.94; N, 10.56%. Found: C, 62.99; H, 5.12; N, 10.33%.
2-[(7-Acetyl-4-cyano-1,6-dimethyl-6-hydroxy-8-(3-nitrophenyl)-5,6,7,8-tetrahydro-isoquinolin-3-yl)thio]- N -(4-tolyl)acetamide (5c). It was synthesized by reaction of 2a with N-(4-tolyl)-2-chloroacetamide (4c).Yield: 95%; m.p.: 187-188°C. IR: 3559 (O-H); 3317 (N-H); 3034 (C-H, sp2); 2973, 2924 (C-H, sp3); 2213 (C≡N); 1701 (C=O, acetyl); 1675 (C=O, amide). 1H NMR: δ 10.12 (s, 1H, NH), 8.06-8.08 (d, 1H, ArH), 7.94-7.95 (m, 1H, ArH), 7.53-7.55 (m, 2H, Ar-H), 7.38-7.40 (d, 2H, ArH ), 7.04-7.06 (d, 2H, ArH), 4.99 (s, 1H, OH), 4.76-4.78 (d, J = 10.0, 1H, C8H), 4.06-4.15 (dd, J = 15, 2H, SCH2), 2.89-3.32 (m, 3H: C7H and C5H2), 2.21 (s, 3H, CH3 of 4-tolyl residue), 2.17 (s, 3H, COCH3), 1.99 (s, 3H, CH3), 1.28 (s, 3H, CH3). 13C NMR: δ 208.74, 200.27, 181.20, 165.58, 160.36, 157.54, 150.02, 147.75, 145.84, 136.23, 134.97, 132.03, 130.00, 128.88, 128.47, 122.54, 121.56, 118.87, 114.90, 103.87, 67.23, 65.74, 55.86, 43.11, 42.28, 34.55, 30.84, 27.33, 24.51, 20.21, 18.36. Anal. Calcd. Anal. calcd for C29H28N4O5S (544.18): C, 63.95; H, 5.18; N, 10.29%. Found: C, 64.04; H, 4.92; N, 9.91%.
2-[(7-Acetyl-4-cyano-1,6-dimethyl-6-hydroxy-8-(3-nitrophenyl)-5,6,7,8-tetrahydro-isoquinolin-3-yl)thio]- N -(4-chlorophenyl)acetamide (5d): It was synthesized by reaction of 2a with N-(chlorophenyl)-2-chloroacetamide (4d). Yield: 84%; m.p.: 205-206°C. IR: 3536 (O-H); 3289 (N-H); 3074 (C-H, sp2); 2973, 2924 (C-H, sp3); 2216 (C≡N); 1694 (C=O, acetyl); 1666 (C=O, amide). 1H NMR: δ 10.37 (s, 1H, NH), 8.06 (d, 1H, ArH), 7.94 (s, 1H, ArH), 7.54-7.56 (m, 4H, ArH), 7.29-7.31 (d, J = 10, 2H, ArH), 4.99 (s, 1H, OH), 4.76-4.78 (d, J = 10, 1H C8H), 4.14-4.17 (dd, 2H, SCH2), 3.30-3.32 (d, J =10, 1H, C5H), 2.93-2.95 (m, 2H: C7H and C5H), 2.17 (s, 3H, COCH3), 1.89 (s, 3H, CH3), 1.28 (s, 3H, CH3). 13C NMR: δ 204.15, 161.49, 155.77, 152.85, 145.47, 143.16, 141.23, 133.10, 130.38, 125.41, 123.93, 123.84, 122.10, 117.96, 116.97, 115.78, 110.30, 99.30, 62.66, 61.15, 38.53, 37.70, 30.01, 26.27, 22.75, 19.90. Anal. Calcd. for C28H25ClN4O5S (564.12): C, 59.52; H, 4.46; N, 9.92%. Found: C, 59.31; H, 4.50; N, 10.13%.
2-[(7-Acetyl-4-cyano-1,6-dimethyl-6-hydroxy-8-(4-nitrophenyl)-5,6,7,8-tetrahydro-isoquinolin-3-yl)thio]acetamide (5e): It was synthesized by reaction of 2b with 2-chloroacetamide (4a).Yield: 88%; m.p.: 178-179°C. IR: 3466, 3355 (O-H, NH2); 2968, 2919 (C-H, sp3); 2222 (C≡N); 1709 (C=O, acetyl); 1662 (C=O, amide).1H NMR: δ 8.09-8.11 (d, J = 10.0, 2H, ArH), 7.54 (s, 1H, NH), 7.30-7.32 (dd, J = 5.0, 2H, ArH), 7.09 (s, 1H, NH), 5.00 (s, 1H, OH), 4.70-4.72 (d, J = 10.0, 1H, C8H), 3.81-3.89 (dd, J = 15.0, 2H, SCH2), 3.25-3.28 (d, J = 15.0, 1H, C5H), 2.88-2.90 (d, J = 10.0, 1H, C7H), 2.83-2.87 (d, J = 20.0, 1H, C5H), 2.23 (s, 3H, COCH3), 1.91 (s, 3H, CH3), 1.24 (s, 3H, CH3). Anal. calcd for C22H22N4O5S (454.13): C, 58.14; H, 4.88; N, 12.33%. Found: C, 57.92; H, 4.59; N, 12.52%.
2-[(7-Acetyl-4-cyano-1,6-dimethyl-6-hydroxy-8-(4-nitrophenyl)-5,6,7,8-tetrahydro-isoquinolin-3-yl)thio]- N -phenylacetamide (5f): It was synthesized by reaction of 2b with N-phenyl-2-chloroacetamide (4b). Yield: 92%; m.p.: 137-138°C. IR: 3525 (O-H); 3322 (N-H); 3061 (C-H, sp2); 2994, 2935 (C-H, sp3); 2217 (C≡N); 1702 (C=O, acetyl); 1687 (C=O, amide).1H NMR: δ 10.21 (s, 1H, NH); 8.09-8.11 (d, J = 10.0, 2H, ArH); 7.50-7.52 (d, J = 10, 2H, ArH); 7.32-7.33 (d, J = 5, 2H, ArH); 7.23-7.24(d, J = 5, 2H, ArH); 6.98-7.01 (t, J = 5, 1H, ArH); 4.98 (s, 1H, OH); 4.73-4.75 (d, J =10,1H, C8H); 4.08-4.16 (dd, 2H, SCH2); 3.28-3.30 (d, J =10, 1H, C5H), 2.89-2.94 (m,, 2H: C7H and C5H); 2.16 (s, 3H, COCH3); 1.89 (s, 3H, CH3); 1.28 (s, 3H, CH3). 13C NMR: δ 208.52, 166.01, 160.51, 157.73, 151.77, 150.03, 146.07, 138.89, 129.52, 128.67, 123.78, 123.26, 119.05, 115.01, 104.03, 78.72, 67.41, 65.71, 56.02, 42.70, 34.74, 31.07, 27.48, 24.50,18.50. Anal. Calcd. for C28H26N4O5S (530.16): C, 63.38; H, 4.94; N, 10.56%. Found: C, 62.99; H, 5.09; N, 10.53%.
2-[(7-Acetyl-4-cyano-1,6-dimethyl-6-hydroxy-8-(4-nitrophenyl)-5,6,7,8-tetrahydro-isoquinolin-3-yl)thio]- N -(4-tolyl)acetamide (5g). It was synthesized by reaction of 2b with N-(4-tolyl)-2-chloroacetamide (4c).Yield: 90%; m.p.: 234-235°C. IR: 3456 (O-H); 3297 (N-H); 3107 (C-H, sp2); 2970 (C-H, sp3); 2218 (C≡N); 1702 (C=O, acetyl); 1682 (C=O, amide). 1H NMR: δ 10.12 (s, 1H, NH), 8.07-8.09 (d, J = 10, 2H, ArH), 7.28-7.36 (m, 4H, ArH), 7.01-7.04 (d, 2H, ArH), 4.99 (s, 1H, OH), 4.68-4.71 (d, 1H, C8H), 4.05-4.07(m, 2H, SCH2), 3.25-33.28(d, 1H, C5H), 2.86-2.88 (m, 2H: C7H and C5H), 2.18 (s, 3H, CH3 of 4-tolyl residue), 2.13 (s, 3H, COCH3), 1.84 (s, 3H, CH3), 1.24 (s, 3H, CH3). Anal. Calcd. for C29H28N4O5S (544.62): C, 63.95; H, 5.18; N, 10.29%. Found: C, 64.08; H, 4.91; N, 9.93%.
2-[(7-Acetyl-4-cyano-1,6-dimethyl-6-hydroxy-8-(4-nitrophenyl)-5,6,7,8-tetrahydro-isoquinolin-3-yl)thio]- N -(4-chlorophenyl)acetamide (5h). It was synthesized by reaction of 2b with N-(4-chlorophenyl)-2-chloroacetamide (4d).Yield: 94%; m.p.: 144-145°C. IR: 3563 (O-H), 3344 (N-H); 3134 (C-H, sp2); 2971, 2937 (C-H, sp3); 2221 (C≡N); 1705 (C=O, acetyl); 1681 (C=O, amide). 1H NMR: δ 10.35 (s, 1H, NH), 8.08-8.11 (m, 2H, ArH), 7.60-7.62 (d, 2H, ArH), 7.29-7.54 (m, 4H, ArH), 4.98 (s, 1H, OH), 4.71-4.73 (d, J = 10, 1H, CH at C-8), 4.06-4.14 (dd, J =15.0, 2H, SCH2), 3.42-3.44 (d, J = 7.0, 1H, C5H), 2.90-2.92 (m,, 2H: C7H and C5H), 2.15(s, 3H, COCH3), 1.85 (s, 3H, CH3), 1.27 (s, 3H, CH3).13C NMR: δ 208.53, 166.23, 164.75, 160.47, 157.63, 151.75, 150.04, 146.07, 137.85, 129.52, 128.73, 128.60, 126.83, 123.77, 120.90, 120.53, 114.98, 103.98, 67.39, 65.71, 55.99, 43.21, 42.65, 34.72, 31.02, 27.46, 24.45, 18.50. Anal. Calcd. for C28H25ClN4O5S (564.12): C, 59.52; H, 4.46; N, 9.92%. Found: C, 59.20; H, 4.67; N, 10.07%.
2-[(7-Acetyl-4-cyano-1,6-dimethyl-6-hydroxy-8-(4-nitrophenyl)-5,6,7,8-tetrahydro-isoquinolin-3-yl)thio]- N -(4-acetylphenyl)acetamide (5i): It was synthesized by reaction of 2a with N-(4-acetylphenyl)-2-chloroacetamide (4e). Yield: 86%; m.p.: 193-194°C. IR: 3540 (O-H); 3337(N-H); 3109 (C-H, sp2); 2968 (C-H, sp3); 2220 (C≡N); 1683 (3 C=O); 1595 (C=N).1H NMR: δ 10.57 (s, 1H, NH), 8.06-8.11 (d, 2H, ArH), 7.84-7.86 (d, 2H, ArH), 7.62-7.65 (d, 2H, ArH), 7.28-7.31 (d, 2H, ArH), 5.02 (s, 1H, OH), 4.76-4. 78 (d, 1H, C8H), 4.36-4.38 (d, 1H, C5H), 4.11-4.13 (dd, 2H, SCH2), 2.88-2..91 (m, 2H: C7H and C5H), 2.12 (s, 3H,COCH3), 1.80 (s, 3H, COCH3), 1.23 (s, 3H, CH3 attached to pyridine ring), 1.03 (s, 3H, CH3). Anal. Calcd. for C30H28N4O6S (572.17): C, 62.92; H, 4.93; N, 9.78%. Found: C, 63.00; H, 4.85; N, 10.06%.
7-Acetyl-1-amino-2-( N -arylcarbamoyl)-5,8-dimethyl-8-hydroxy-6-(3-nitrophenyl or 4-nitrophenyl)-6,7,8,9-tetrahydrothieno[2,3- c ]isoquinolines 6b-d,f,g; general methods.
Method A)
To a suspension of 5b-d,f,g (10 mmol) in abs. ethanol (60 mL), anhydrous sodium carbonate (0.30 g)) was added. The reaction mixture was refluxed for 3 hours. The yellow crystals that formed while hot were collected, washed with water, dried in air and then recrystallized from dioxane to give 6b-d,f,g, repectively.
7-Acetyl-1-amino-5,8-dimethyl-8-hydroxy-6-(3-nitrophenyl)- N -phenyl-6,7,8,9-tetrahydrothieno[2,3-c]isoquinoline-2-carboxamide (6b).
Yield: 87%; m.p.: 287-288°C. IR: 3415, 3388, 3314 (O-H, NH2, N-H); 2914 (C-H, sp3); 1703 (C=O, acetyl); 1622 (C=O, amide).1H NMR: δ 9.43 (s, 1H, NH); 7.31-7.84 (m, 9H, ArH); 7.09 (s, 2H, NH2); 4.86-4.88 (d, J =10,1H, C6H); 4.84 (s, 1H, OH); 3.64-3.67(d, J =15, 1H, C9H), 3.41-3.44 (d, J =15, 1H, C7H); 2.92-2.94 (d, J =10, 1H, C9H); 2.21 (s, 3H, COCH3); 2.03 (s, 3H, CH3); 1.33 (s, 3H, CH3). 13C NMR: δ 209.44, 164.31, 158.22, 156.58, 149.38, 147.92, 147.07, 142.88, 138.83, 135.08, 130.11, 128.36, 128.24, 123.45, 123.02, 122.40, 121.51, 121.26, 97.03, 67.14, 65.90, 42.90, 41.98, 31.17, 27.94, 24.74. Anal. Calcd. for C28H26N4O5S (530.16): C, 63.38; H, 4.94; N, 10.56%. Found: C, 62.99; H, 5.12; N, 10.46%.
7-Acetyl-1-amino-5,8-dimethyl-8-hydroxy-6-(3-nitrophenyl)- N -(4-tolyl)-6,7,8,9-tetrahydrothieno[2,3- c ]isoquinoline-2-carboxamide (6c). Yield: 92%; m.p.: 291-292°C. IR: 3418, 3386, 3313 (O-H, NH2, N-H); 3075 (C-H, sp2); 2914(C-H, sp3); 1706 (C=O, acetyl); 1624 (C=O, amide).1H NMR: δ 9.35 (s, 1H, NH); 7.06-8.08 (d, J = 10, 1H, ArH); 7.84 (s, 1H, ArH); 7.53-7.58 (m, 4H, ArH); 7.12-7.14 (d, 2H, J = 10, ArH); 7.07 (s, 2H, NH2); 4.86-4.88 (d, J =10, 1H, C6H); 4.84 (s, 1H, OH); 3.64-3.67(d, J =15, 1H, C9H), 3.41-3.45 (d, J = 20, 1H, C7H); 2.93-2.95 (d, J =10,1H, C9H); 2.28 (s, 3H, CH3 of 4-tolyl residue); 2.21 (s, 3H, COCH3); 2.03 (s, 3H, CH3); 1.33 (s, 3H, CH3).13C NMR: δ 209.44, 164.19, 158.12, 156.53, 149.19, 147.92, 147.08, 142.83, 136.25, 135.07, 132.42, 130.11, 128.77, 128.21, 123.08, 122.40, 121.51, 121.31, 97.20, 67.15, 65.90, 42.91, 41.97, 31.18, 27.95, 24.73, 20.46. Anal. Calcd. for C29H28N4O5S (544.18): C, 63.95; H, 5.18; N, 10.29%. Found: C, 64.14; H, 4.92; N, 9.93%.
7-Acetyl-1-amino- N -(4-chlorophenyl)-5,8-dimethyl-8-hydroxy-6-(3-nitrophenyl)-6,7,8,9-tetrahydrothieno[2,3- c ]isoquinoline-2-carboxamide (6d). It was obtained by cyclization of compound 8c. Yield: 94%; m.p.: 293-294°C. IR: 3417, 3383, 3314 (O-H, NH2, N-H); 3095 (C-H, sp2); 2967, 2916(C-H, sp3); 1706 (C=O, acetyl); 1622 (C=O, amide).1H NMR: δ 9.56 (s, 1H, NH); 7.36-8.08 (m, 8H, ArH); 7.13 (s, 2H, NH2); 4.86-4.88 (d, J =10,1H, C6H); 4.85 (s, 1H, OH); 3.64-3.67 (d, J =15, 1H, C9H), 3.40-3.44 (d, J =20, 1H, C7H); 2.93-2.95 (d, J =10,1H, C9H); 2.21 (s, 3H, COCH3); 2.04 (s, 3H, CH3); 1.33 (s, 3H, CH3).13C NMR: δ 209.42, 164.35, 158.33, 156.65, 149.62, 147.92, 147.04, 142.94, 135.07, 130.10, 128.27, 128.23, 126.96, 122.95, 122.65, 122.41, 121.51, 96.81, 67.14, 65.88, 42.8, 41.99, 31.17, 27.94, 24.74. Anal. Calcd. for C28H25ClN4O5S (564.12): C, 59.52; H, 4.46; N, 9.92%. Found: C, 59.69; H, 4.41; N, 10.16%.
7-Acetyl-1-amino-5,8-dimethyl-8-hydroxy-6-(4-nitrophenyl)-N-phenyl-6,7,8,9-tetrahydrothieno[2,3-c]isoquinoline-2-carboxamide (6f). Yield: 91%; m.p.: 285-286°C. IR: 3406, 3320 (O-H, NH2, N-H); 2921(C-H, sp3); 1703 (C=O, acetyl); 1622 (C=O, amide).1H NMR: δ 9.41 (s, 1H, NH); 8.11-8.13 (d, J =10, 2H, ArH); 7.67-7.69 (d, J =10, 2H, ArH); 7.28-7.33 (m, 5H, ArH); 7.08 (s, 2H, NH2); 4.84 (s, 1H, OH); 4.82-4.84 (d, J =10, 1H, C6H); 3.59-3.63(d, J =20, 1H, C9H), 3.40-3.43 (d, J =15, 1H, C7H); 2.87-2.89 (d, J =10,1H, C9H); 2.19 (s, 3H, COCH3); 2.00 (s, 3H, CH3); 1.32 (s, 3H, CH3). 13C NMR: δ 209.25, 164.33, 158.17, 156.61, 152.92, 149.35, 145.94, 142.71, 138.84, 129.40, 128.37, 128.22, 123.80, 123.46, 123.02, 121.26, 97.03, 67.14, 65.73, 43.19, 41.96, 31.19, 27.92, 24.61. Anal. Calcd. for C28H26N4O5S (530.16): C, 63.38; H, 4.94; N, 10.56%. Found: C, 62.98; H, 5.01; N, 10.62%.
7-Acetyl-1-amino-5,8-dimethyl-8-hydroxy-6-(4-nitrophenyl)- N -(4-tolyl)-6,7,8,9-tetrahydrothieno[2,3- c ]isoquinoline-2-carboxamide (6g). Yield: 92%; m.p.:292- 293°C. IR: 3400, 3322 (O-H, NH2, N-H); 2919 (C-H, sp3); 1701 (C=O, acetyl); 1623 (C=O, amide).1H NMR: δ 9.33 (s, 1H, NH); 8.11-8.13 (d, J =10, 2H, Ar-H); 7.55-7.57 (d, J =10, 2H, ArH); 7.27-7.29 (d, J =10, 2H, ArH); 7.11-7.13 (d, J =10, 2H, ArH); 7.05 (s, 2H, NH2); 4.84 (br s, 1H, OH); 4.82-4.84 (d, J =10, 1H, C6H); 3.59-3.62 (d, J =15, 1H, C9H), 3.40-3.44 (d, J = 20, 1H, C7H); 2.86-2.89 (d, J =15, 1H, C9H); 2.27 (s, 3H, CH3 of 4-tolyl residue); 2.19 (s, 3H, COCH3); 2.01 (s, 3H, CH3); 1.32 (s, 3H, CH3). 13C NMR: δ 209.25, 164.19, 158.06, 156.55, 152.92,149.14, 145.76, 142.65, 136.26, 129.38, 128.77, 128.17, 123.79, 123.06, 121.30, 97.19, 67.13, 65.74, 43.18, 41.94, 31.18, 27.92, 24.59, 20.44. Anal. Calcd. for C29H28N4O5S (544.18): C, 63.95; H, 5.18; N, 10.29%. Found: C, 64.13; H, 4.92; N, 9.99%.
Method B).
To mixture of 2a,b (10 mmol) and respective N-aryl-2-chloroacetamide 4b-d (10 mmol) in ethanol (60 mL), anhydrous sodium carbonate (1.30 g) was added. The resulting mixture was refluxed for 3 hours. The solid that formed while hot was collected, washed with water, dried in air and then recrystallized from dioxane to give compounds 6b-d,f,g; yield: 80-86%.
Cytotoxic activty
The cytotoxicity activity of the some synthesized compounds was determined according to the MTT method [26–28].
The pancreatic (PACA2) and human cancer lung (A549) cells were cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum and 1% GlutaMAX. Then the cells were seeded into sterile 96-well plates at a density of 10 × 103 cells/well and maintained at 37°C for 24 h. Cancerous cells were exposed to compounds at concentrations of 0.75, 1.75, 3.125, 6.250, 12.500, 25, 50, and 100 µM for 72 h. The media was removing and add 40µl MTT stock solution to each well. The resulting solutions were incubated for more than 4 h. Subsequently, then add 120 µL of 10% SDS as solubilising reagent. SPSS Software program was used to calculate the IC50 and IC50 ranges.
Antioxidant activity
DPPH has been used for measurement of free radical scavenging ability of antioxidants. Reduction of an alcoholic DPPH solution [29–31] in the presence of a hydrogen-donating antioxidant is the mainly step of this method. Hydrogen atom or electron-donation ability of the tested compounds were measured spectrophotometrically from the bleaching of the purple-colored ethanol solution of 2,2-diphenyl-1-picrylhydrazyl (DPPH). In this study, antioxidant activity of the tested compounds was measured using the stable radical 2,2- diphenyl-1-picrylhydraziyl (DPPH). The free radical scavenging capacity of the tested compounds was determined using DPPh. A solution 1: prepared by dissolving DPPH (0.002 gm) in ethanol (50 mL etnanol ). Solution 2: prepared by dissolving different weights 0.1, 0.05, 0.01 grams of each sample in 1mL of DMSO then take 10 µL of each sample solution with 1mL ethanol. Then mix 1mL of solution 1 with 1mL of solution 2 and the resulting mixture was vortexed thoroughly and left in the dark for about 30 min. The absorbance of the mixture was spectrophotometrically measured at λmax = 517 nm against blank 1mL absolute ethanol and compared to the ascorbic acid (Vitamin C). DPPH radical scavenging activity (% RSA) of compounds was calculated from the absorbance at the start (0) and after some reaction time (T) according to the equation (1).
(% RSA) = (ABS-ATS) / ABS x 100 (1)
Where ABS is the absorbance of blank sample (DPPH) solution without the compound to be tested and ATS is the absorbance of tested sample.