N-arylalkylamines as plasmodione analogues is described. Using the naturally occurring and commercially available menadione as starting material, a 2-step sequence using a Kochi-Anderson reaction and subsequent Pd-catalyzed Suzuki-Miyaura coupling was developed to prepare three representative biphenyl derivatives in good yields for antimalarial evaluation. In addition, synthetic methodologies to afford 3-benzylmenadione derivatives bearing a terminal -N(Me)2 or -N(Et)2 in different positions  on the aryl ring of the benzylic chain of plasmodione were investigated through reductive amination was used as the optimal route to prepare these protonable N-arylalkylamine privileged scaffolds. The antimalarial activities were evaluated and discussed in light of their physicochemical properties. Among the newly synthesized compounds, the para-position of the substituent remains the most favourable position on the benzyl chain and the carbamate -NHBoc was found active both in vitro (42 nM versus 29 nM for plasmodione) and in vivo in Plasmodium berghei-infected mice. The measured acido-basic features of these new molecules support the cytosol-food vacuole shuttling properties of non-protonable plasmodione derivatives essential for redox-cycling. These findings may be useful in antimalarial drug optimization.With the aim of increasing the structural diversity on the early antimalarial drug plasmodione, an efficient and versatile procedure to prepare a series of biaryl- and  Plasmodium in tropical and subtropical areas and is curable if treated promptly and adequately. The most dangerous and harmful species, P. falciparum is responsible for malignant malaria with complications such as cerebral malaria or severe anaemia, and is responsible for significant mortality and morbidity including the death of approximately 0.6 million people worldwide by year, mostly African children younger than 5 years + calcd. for C10H12O3N, 194.0817; found, 194.0770.rocedure , 4-amino2-(3-Acetamidophenyl)acetic Acid (15). 3-Aminophenylacetic acid  was added to acetic acid (2 mL) under microwave irradiation (150 °C) for 10 min. The reaction mixture was poured into a mixture of ethyl acetate (1:50), then washed with water and then allowed to dry to yield the compound 15 : δ = 12.27 , 9.90 , 7.47 , 7.24–7.19 , 6.91 , 3.51 , 2.03 . 13C-NMR : δ = 172.52 (Cq), 168.21 (Cq), 139.26 (Cq), 135.39 (Cq), 128.48 (Cq), 124.00 (Cq), 119.76 (Cq), 117.32 (Cq), 40.86 (CH2), 23.94 (CH3). IR: 3298 (w), 1699 (s), 1662 (s), 1544 (m), 1492 (m), 1411 (m), 1308 (m), 1273 (m), 1232 (m), 886 (m), 782 (s), 714 (s).pound 15  acetic Acid (21a). Oxindole 20  was diluted in 6 M HCl (15 mL) and heated to reflux with stirring overnight. The solution was cooled and washed with dichloromethane (3 × 100 mL). The aqueous phase was concentrated to give 21 as a white solid . The crude product was involved directly in the next step. A solution of 2-aminophenylacetic acid hydrochloride  in dichloromethane (110 mL) at room temperature was treated with triethylamine  and acetyl chloride . The reaction mixture was stirred at r.t. for 3 h. The reaction mixture was diluted with EtOAc and washed with water. The aqueous layer was extracted with ethyl acetate. The combined extracts were rinsed with brine, dried over MgSO4. Purification on silica gel with cyclohexane/ethyl acetate (20:5 to 1:1) gives the desired product 21a : δ = 8.21 , 7.34–7.25 , 7.20–7.15 , 3.71 , 2.67 . 13C-NMR : δ = 175.47 (Cq), 171.03 (Cq), 141.54 (Cq), 128.37 (CH), 125.09 (CH), 124.10 (CH), 123.57 (Cq), 116.82 (CH), 36.80 (CH2), 26.83 (CH3).tert-Butylmethyl)phenyl) carbamate (6). A solution of menadione  and phenylacetic acid 5  in acetonitrile (27 mL) and water (9 mL) was heated to 70 °C. AgNO3  was added. (NH4)2S2O8  in acetonitrile (3 mL) and water (1 mL) was added dropwise over a period of 45 min and then heated at reflux for 2 h. The acetonitrile was removed in vacuo. The product was extracted with EtOAc (1 × 10 mL), dichloromethane (4 × 10 mL), dried over MgSO4 and purified by chromatography on silica gel (cyclohexane/dichloromethane 30:5) to give the pure compound 6 : δ = 8.05–8.08 , 7.66–7.71 , 7.23 , 7.12 , 6.36 , 3.95 , 2.21 , 1.47 . 13C-NMR : δ = 185.42 (Cq), 184.68 (Cq), 152.73 (Cq), 145.38 (Cq), 144.21 (Cq), 136.70 (Cq), 133.47 (CH), 133.44 (CH), 132.61 (Cq), 132.10 (Cq), 132.03 (Cq), 129.14 (CH), 126.45 (CH), 126.25 (CH), 118.88 (CH), 80.49 (Cq), 31.74 (CH2), 28.30 (CH3), 13.22 (CH3). EI MS ): 377.2 , 321.1 (66), 305.9 (100), 261.1 (59), 201.3 (14), 160.1 (18), 121.1 (21). IR: 3439 , 1704 (w), 1685 (w), 1660 (s), 1618 (m), 1596 (m), 1521 (m), 1370 (w), 1315 (m), 1296 (m), 1236 (w), 1162 (s), 709 (w). EA: obs. C, 72.94%; H, 6.16%; N, 3.74%, calcd. C, 73.19%; H, 6.14%, N, 3.71% for C23H23NO4.tert-Butyl methyl)phenyl) carbamate (14). A solution of menadione  and phenylacetic acid 13  in acetonitrile (36 mL) and water (12 mL) was heated to 70 °C. AgNO3  was added. (NH4)2S2O8  in acetonitrile (11 mL) and water (3.5 mL) was added dropwise over a period of 45 min and then heated at reflux for 3 h. The acetonitrile was removed in vacuo. The product was extracted with EtOAc (2 × 10 mL), dichloromethane (4 × 10 mL), dried over MgSO4 and purified by chromatography on silica gel (toluene) a second chromatography on silica was necessary (cyclohexane/dichloromethane 10:1) to give the pure compound 14 : δ = 8.10–8.05 , 7.72–7.66 , 7.28–7.26 , 7.20–7.14 , 6.86 , 6.46 , 3.99 , 2.24 , 1.49 . 13C-NMR : δ = 185.45 (Cq), 184.73 (Cq), 152.73 (Cq), 145.24 (Cq), 144.70 (Cq), 139.04 (Cq), 138.76 (Cq), 133.58 (CH), 133.57 (CH), 132.23 (Cq), 132.12 (Cq), 129.37 (CH), 126.60 (CH), 126.38 (CH), 123.31 (CH), 118.81 (CH), 116.79 (CH), 80.60 (CH), 32.48 (CH2), 28.43 (CH3), 13.44 (CH3). HRMS-ESI (m/z): [M + Na]+ calcd. for C23H23NO4Na 400.1519; found 400.1539.N-methyl)phenyl)acetamide (9). A solution of menadione  and phenylacetic acid 8  in acetonitrile (12 mL) and water (4 mL) was heated to 70 °C. AgNO3  was added. (NH4)2S2O8  in 4 mL acetonitrile and 2 mL water was added dropwise over a period of 45 min and then heated at reflux for 3 h. The acetonitrile was removed in vacuo. The product was extracted with dichloromethane (4 × 10 mL), dried over MgSO4 and purified by chromatography on silica gel (cyclohexane/ethyl acetate 80:20 to cyclohexane/ethyl acetate 50:50) to give the pure compound 9 : δ = 8.12–8.05 , 7.72–7.67 , 7.38 , 7.17 , 3.99 , 2.24 , 2.14 . 13C-NMR : δ = 185.50 (Cq), 184.80 (Cq), 168.28 (Cq), 145.38 (Cq), 144.49 (Cq), 136.39 (Cq), 134.13 (Cq), 133.64 (CH), 132.27 (Cq), 132.17 (Cq), 129.29 (CH), 126.60 (CH), 126.44 (CH), 120.36 (CH), 32.01 (CH2), 24.67 (CH3), 13.40 (CH3); IR: 3359 (w), 1690 (s), 1661 (s), 1648 (s), 1593 (m), 1533 (s), 1411 (w), 1371 (m), 1334 (m), 1311 (s), 1295 (vs), 1259 (m), 972 (w), 818 (m), 735 (m), 709 (s), 679 (w).N-methyl)phenyl)acetamide (16). A solution of menadione  and phenylacetic acid 15  in acetonitrile (6 mL) and water (2 mL) was heated to 70 °C. AgNO3  was added. (NH4)2S2O8  in acetonitrile (1.5 mL) and water (0.5 mL) was added dropwise over a period of 45 min and then heated at reflux for 3 h. The acetonitrile was removed in vacuo. The product was extracted with dichloromethane (4 × 10 mL), dried over MgSO4 and purified by chromatography on silica gel (cyclohexane/ethyl acetate 10:1) to give the pure compound 16 : δ = 8.10–8.03 , 7.71–7.65 , 7.41 , 7.32 , 7.20 , 6.96 , 3.99 , 2.24 , 2.12 . 13C-NMR : δ = 185.41 (Cq), 184.79 (Cq), 168.41 (Cq), 145.15 (Cq), 144.80 (Cq), 139.09 (Cq), 138.36 (Cq), 133.63 (CH), 132.25 (Cq), 132.12 (Cq), 129.41 (CH), 126.59 (CH), 126.43 (CH), 124.59 (CH), 119.99 (CH), 118.16 (CH), 32.50 (CH2), 24.71 (CH3), 13.44 (CH3); IR: 1661 (m), 01609 (w), 1551 (w), 1490 (w), 1374 (w), 1319 (w), 1293 (m), 726 (w).N-methyl)phenyl)acetamide (22). A solution of 2-aminophenylacetic acid hydrochloride  in dichloromethane (110 mL) at room temperature was treated with triethylamine  and acetyl chloride . The reaction mixture was stirred at r.t. for 3 h. The reaction mixture was diluted with EtOAc and washed with water. The aqueous layer was extracted with ethyl acetate. The combined extracts were rinsed with brine, dried over MgSO4. Thus, a solution of the crude and menadione  in acetonitrile (32 mL) and water (11 mL) was heated to 70 °C. AgNO3  was added. (NH4)2S2O8  in acetonitrile (10 mL) and water (5 mL) was added dropwise over a period of 45 min and then heated at reflux for 3 h. The acetonitrile was removed in vacuo. The product was extracted with dichloromethane (4 × 10 mL), dried over MgSO4 and purified by chromatography on silica gel (cyclohexane/ethyl acetate 10:1 to cyclohexane/ethyl acetate 5:5) to give the pure compound 22 : δ = 8.99 , 8.11–8.05 , 7.86 , 7.73–7.70 , 7.25–7.19 , 7.07–7.02 , 3.95 , 2.43 , 2.31 . 13C-NMR : δ = 186.32 (Cq), 184.90 (Cq), 168.86 (Cq), 145.77 (Cq), 145.07 (Cq), 135.98 (Cq), 134.12 (CH), 133.70 (CH), 132.10 (Cq), 131.81 (Cq), 129.95 (CH), 129.24 (Cq), 127.72 (CH), 126.63 (CH), 125.08 (CH), 124.74 (CH), 28.43 (CH2), 24.39 (CH3), 14.14 (CH3). IR: 3394 (w), 1674 (m), 1660 (s), 1505 (m), 1467 (m), 1295 (vs), 794 (m), 762 (s), 716 (s), 690 (m).2-Methyl-3-(4-bromo-benzyl)-4a,8a-dihydro-naphthoquinone (25c). A solution of menadione (5.81 mmol) and a phenylacetic acid derivative 24c (11.58 mmol) in a mixture of (52.5 mL of acetonitrile and 17.5 mL of water) was heated to 85 °C. Then AgNO3  was added and (NH4)2S2O8  in 15 mL of acetonitrile and 5 mL of water was added dropwise over a period of 45 min and then heated at reflux for two hours. The acetonitrile was removed in vacuo. The aqueous phase was extracted with dichloromethane (4 × 10 mL), dried over MgSO4 and purified by flash-chromatography on silica gel (petroleum ether/dichloromethane 1:1) to give the compound 25c : δ = 8.03–8.10 , 7.66–7.71 , 7.36 , 7.09 , 3.96 , 2.22 . 13C-NMR : δ = 185.20 (Cq), 184.54 (Cq), 144.75 (Cq), 144.57 (Cq), 137.06 (Cq), 133.58 (CH), 132.08 (Cq), 131.94 (Cq), 131.71 (CH), 130.32 (CH), 126.50 (CH), 126.35 (CH), 120.31 (Cq), 31.93 (CH2), 13.31 (CH3). EI MS ): 340.1 , 325.0 (100), 246.1 (63), 215.1 (41), 202.1 (49), 128.1 (72), 76.0 (74). IR (KBr): 3449 cm−1 , 3068 (w), 2962 (w), 1661 (vs), 1624 (m), 1618 (m), 1594 (s), 1486 (s), 1376 (m), 1332 (s), 1315 (s), 1294 (vs), 1071 (m), 1010 (s), 971 (w), 815 (m), 787 (s), 730 (m), 702 (m), 629 (w), 426 (w). EA: obs. C, 63.02%; H, 3.84%, calcd. C, 63.36%; H, 3.84% for C18H13BrO2.2-Methyl-3-(2-bromo-benzyl)-4a,8a-dihydro-naphthoquinone (25a). After chromatography on silica gel (petroleum ether/dichloromethane 1:1), 1.75 g  of 25a : δ = 8.06–8.11 , 7.66–7.72 , 7.56 , 7.13 , 7.04 , 6.89 , 4.11 , 2.10 . 13C-NMR : δ = 184.98 (Cq), 184.30 (Cq), 145.85 (Cq), 144.53 (Cq), 137.31 (Cq), 133.55 (CH), 133.53 (CH), 132.87 (CH), 132.13 (Cq), 131.96 (Cq), 128.59 (CH), 127.93 (CH), 127.55 (CH), 126.53 (CH), 126.33 (CH), 124.67 (Cq), 32.65 (CH2), 13.26 (CH3). EI MS ): 261.1 , 231.1 (11), 202.1 (11), 130.1 (8), 76.0 (10). IR: 3441 cm−1 , 3068 (w), 3017 (w), 2923 (w), 1660 (vs), 1621 (s), 1594 (s), 1467 (m), 1439 (m), 1376 (w), 1318 (m), 1296 (vs), 1263 (m), 1223 (w), 1184 (w), 1025 (m), 976 (m), 787 (w), 749 (s), 729 (m), 695 (w), 663 (w). EA: obs. C, 63.12%; H, 3.91%; Br, 23.31%, calcd. C, 63.36%; H, 3.84%; Br, 23.42% for C18H13BrO2.) of 25a  were iso2-Methyl-3-(3-bromo-benzyl)-4a,8a-dihydro-naphthoquinone (25b). After chromatography on silica gel (petroleum ether/dichloromethane 1:1), 328 mg  of 25b : δ = 7.98–8.02 , 7.59–7.65 , 7.28 , 7.24 , 7.02–7.09 , 3.92 , 2.16 . 13C-NMR : δ = 185.16 (Cq), 184.45 (Cq), 144.78 (Cq), 144.50 (Cq), 140.35 (Cq), 133.59 (CH), 132.10 (Cq), 131.94 (Cq), 131.52 (CH), 130.16 (CH), 129.65 (CH), 127.26 (CH), 126.54 (CH), 126.36 (CH), 122.72 (Cq), 32.09 (CH2), 13.35 (CH3). EI MS ): 340.0 , 325.06 (100), 246.13 (18), 215.14 (7), 202.12 (8), 184.99 (12), 76.0 (10). IR (KBr): 3430 cm−1 , 1658 (vs), 1620 (vs), 1595 (vs), 1568 (s), 1474 (s), 1431 (m), 1381 (s), 1334 (vs), 1290 (vs), 1261 (s), 1180 (s), 1074 (m), 974 (s), 955 (s), 793 (s), 780 (vs), 728 (vs), 692 (s), 687 (s), 422 (m). EA: obs. C, 63.55%; H, 3.94%; Br, 23.69%, calcd. C, 63.36%; H, 3.84%; Br, 23.42% for C18H13BrO2.) of 25b  were iso2-(4-Aminobenzyl)-3-methylnaphthalene-1,4-dione (7). 4-Acetamide derivative 9  was added to HCl (1 mL) in a mixture of water and methanol  under microwave irradiation (150 °C) for 2 h. The reaction mixture was poured into ethyl acetate then washed with a saturated Na2CO3 solution and then allowed to dry. 172 mg (76%) of compound 7 : δ = 8.09–8.06 , 7.69–7.67 , 7.03 , 6.60 , 3.91 , 2.24 . 13C-NMR : δ = 185.68 (Cq), 184.93 (Cq), 145.99 (Cq), 144.95 (Cq), 143.97 (Cq), 133.53 (CH), 133.49 (CH), 132.27 (Cq), 132.25 (Cq), 129.67 (CH), 127.97 (Cq), 126.54 (CH), 126.33 (CH), 115.50 (CH), 31.69 (CH2), 13.33 (CH3). IR: 3474 (w), 3377 (w), 2932 (w), 1653 (s), 1617 (s), 1590 (m), 1515 (s), 1336 (m), 1293 (vs), 1182 (m), 973 (m) 816 (m), 768 (m), 708 (vs). HRMS-ESI (m/z): [M + H]+ calcd. for C18H16NO2 278.1176; found 278.1154.mpound 7  was isol2-(3-Aminobenzyl)-3-methylnaphthalene-1,4-dione (17). 3-Acetamide derivative 16  was added to HCl (1 mL) in a mixture of water and methanol  under microwave irradiation (150 °C) for 20 min. The reaction mixture was poured into ethyl acetate then washed with a saturated Na2CO3 solution and then allowed to dry. After chromatography on silica gel (cyclohexane/ethyl acetate 75:25 to cyclohexane/ethyl acetate 1:1), 150 mg  of compound 17 : δ = 8.12–8.06 , 7.73–7.67 , 7.05 , 6.62 , 6.56–6.48 , 3.95 , 2.24 . 13C-NMR : δ = 185.67 (Cq), 184.95 (Cq), 146.88 (Cq), 145.63 (Cq), 144.66 (Cq), 139.46 (Cq), 133.69 (CH), 133.37 (CH), 132.40 (Cq), 132.32 (CH), 129.75 (CH), 126.72 (CH), 126.50 (CH), 119.19 (CH), 115.49 (CH), 113.54 (CH), 32.56 (CH2), 13.52 (CH3); ESI MS: 278.12 (M+). IR: 3452 cm−1 (w), 3372 (w), 2921 (w), 1649 (m), 1615 (s), 1588 (s), 1492 (m), 1335 (m), 1292 (vs), 974 (m), 867 (m), 716 (s).pound 17  was isol2-(4-(Dimethylamino)benzyl)-3-methylnaphthalene-1,4-dione (3). Sodium cyanoborohydride  was added by small portions to a stirred solution of 7  in acetonitrile (5 mL) and 37% aqueous formaldehyde  in acetonitrile (1 mL) placed in a cool bath was then added. An additional glacial acetic acid portion  was added and stirring for 2 h. The reaction mixture was poured into diethyl ether (10 mL) and then washed with 1N potassium hydroxide (10 mL) and brine. The organic phase was dried and evaporated in vacuo. The crude product was purified by chromatography on silica gel using a mixture of cyclohexane/ethyl acetate (10:1 to 5:5) affording the compound 3 : δ = 8.09–8.06 , 7.71–7.67 , 7.12 , 6.67 , 3.93 , 2.89 , 2.27 . 13C-NMR : δ = 185.72 (Cq), 185.00 (Cq), 145.96 (Cq), 143.65 (Cq), 142.94 (Cq), 133.51 (CH), 133.47 (CH), 132.28 (Cq), 129.52 (CH), 126.55 (CH), 126.31 (CH), 113.26 (Cq), 40.96 (CH3), 31.56 (CH2), 13.35 (CH3). IR: 2923 cm−1 (w), 2852 (w), 1660 (s), 1615 (m), 1594 (m), 1520 (s), 1291 (vs), 1130 (m), 945 (m), 814 (m), 689 (s).2-(3-(Dimethylamino)benzyl)-3-methylnaphthalene-1,4-dione (18). Sodium cyanoborohydride  was added by small portions to a stirred solution of 17  in acetonitrile (5 mL) and 37% aqueous formaldehyde  in acetonitrile (1 mL) was then added. An additional glacial acetic acid portion  was added and stirring for 2 h. The reaction mixture was poured into diethyl ether (10 mL) and then washed with potassium hydroxide 1 N (10 mL) and brine. The organic phase was dried and evaporated in vacuo. The crude product was purified by chromatography on silica gel using a mixture of cyclohexane/ethyl acetate (10:1 to 5:5) affording the compound 18 : δ = 8.12–8.06 , 7.72–7.66 , 7.12 , 6.65 , 6.60–6.55 , 4.00 , 2.91 , 2.27 . 13C-NMR : δ = 185.58 (Cq), 184.82 (Cq), 150.93 (Cq), 145.74 (Cq), 144.35 (Cq), 138.95 (Cq), 133.51 (CH), 133.46 (CH), 132.26 (Cq), 132.25 (Cq), 129.37 (CH), 126.56 (CH), 126.31 (CH), 116.97 (CH), 113.17 (CH), 110.94 (CH), 40.69 (CH3), 32.90 (CH2), 13.42 (CH3). IR: 2920 cm−1 (w), 2807 (w), 1661 (vs), 1594 (vs), 1501 (m), 1361 (m), 1292 (m), 1292 (vs), 997 (m), 974 (m), 842 (m), 758 (s), 744 (s), 691 (vs).2-(4-(Diethylamino)benzyl)-3-methylnaphthalene-1,4-dione (11). Sodium cyanoborohydride  was added by small portions to a stirred solution of 7  and acetaldehyde  in acetonitrile (1 mL) placed in a cool bath was added. Glacial acetic acid  was then added and stirring for 2 h. The reaction mixture was poured into diethyl ether (10 mL) and then washed with potassium hydroxide 1N (10 mL) and brine. The organic phase was dried and evaporated in vacuo. The crude product was purified by chromatography on silica gel using a mixture of cyclohexane/ethyl acetate (10:1 to 5:5) affording the compounds 10 (17%) and 11 : δ = 8.11–8.04 , 7.71–7.65 , 7.08 , 6.58 , 3.91 , 3.29 , 2.28 , 1.11 ; 13C-NMR : δ = 185.81 (Cq), 185.02 (Cq), 146.60 (Cq), 146.21 (Cq), 143.69 (Cq), 133.48 (Cq), 133.43 (CH), 132.32 (CH), 129.73 (Cq), 129.62 (CH), 126.55 (CH), 126.30 (CH), 124.56 (Cq), 112.25 (CH), 44.48 (CH2), 31.50 (CH2), 13.37 (CH3), 12.71 (CH3). ESI MS: 333.17 (M+); EA: obs. C, 80.19%; H, 8.01%; N, 5.35%, calcd. C, 79.25%; H, 6.25%, N, 4.20% for C22H23NO2; HRMS-ESI (m/z): [M + H]+ calcd. for C22H24NO2 334.1802; found 334.1765.) and 11 ; 36% as 2-(3-(Diethylamino)benzyl)-3-methylnaphthalene-1,4-dione (19). Sodium cyanoborohydride  was added by small portions to a stirred solution of 17  and acetaldehyde  in acetonitrile (1 mL) placed in a cool bath was added. Glacial acetic acid  was then added and stirring for 2 h. The reaction mixture was poured into diethyl ether (10 mL) and then washed with 1N potassium hydroxide (10 mL) and brine. The organic phase was dried and evaporated in vacuo. The crude product was purified by chromatography on silica gel using a mixture of cyclohexane/ethyl acetate (10:1 to 5:5) to afford the compound 19 : δ = 8.12–8.05 , 7.72–7.66 , 7.09 , 6.60 , 6.53–6.47 , 3.99 , 3.31 , 2.29 , 1.13 . 13C-NMR : δ = 185.66 (Cq), 184.87 (Cq), 148.13 (Cq), 145.88 (Cq), 144.16 (Cq), 139.07 (Cq), 133.52 (CH), 133.45 (CH), 132.27 (Cq), 132.24 (Cq), 129.57 (CH), 126.54 (CH), 126.31 (CH), 115.67 (CH), 112.40 (CH), 110.10 (CH), 44.47 (CH2), 32.98 (CH2), 13.45 (CH3), 12.70 (CH3). IR: 2975 cm−1 (w), 2929 (w), 1658 (s), 1595 (vs), 1498 (m), 1290 (vs), 1023 (m), 975 (m), 755 (m), 731 (s), 693 (s). HRMS-ESI (m/z): [M + H]+ calcd. for C22H24NO2 334.1802; found 334.1819.25 , boronic acid derivative , K2CO3  and finally dissolved in dioxane/water (12 mL/3 mL). The solution was degassed by bubbling argon through the mixture for 20 min. Then 4 mol % PdCl2(dppf)  was added, the Schlenk-tube was sealed and heated overnight at 80 °C. The reaction was quenched by adding 10 mL H2O. All volatiles solvents were removed in vacuo, the product was extracted with CH2Cl2, dried over MgSO4 and purified by flash-chromatography.A Schlenk-tube was flushed with argon and successively filled with 2-(4′-Dimethylamino-biphenyl-4-ylmethyl)-3-methyl-naphthoquinone (26). 4-Dimethylamino phenylboronic acid was used as the boronic acid coupling partner for compound 25. After chromatography on silica gel (dichloromethane:petroleum ether/3:1), 96 mg  of 26 : δ = 8.05–8.11 , 7.65–7.71 , 7.43 , 7.24 , 6.76 , 4.03 , 2.96 , 2.27 . 13C-NMR : δ = 185.47 (Cq), 184.74 (Cq), 145.43 (Cq), 144.34 (Cq), 139.43 (Cq), 135.69 (Cq), 133.49 (CH), 133.46 (CH), 132.15 (Cq), 132.09 (Cq), 128.94 (CH), 127.58 (CH), 126.53 (CH), 126.52 (CH), 126.28 (CH), 112.75 (CH), 40.60 (CH3), 32.08 (CH2), 13.37 (CH3). EI MS ): 381.2 , 366.1 (19). IR (KBr): 3432 cm−1 , 3028 (w), 2922 (w), 2855 (w), 2803 (w), 1660 (vs), 1612 (vs), 1595 (s), 1534 (w), 1504 (vs), 1444 (w), 1375 (w), 1357 (m), 1333 (m), 1294 (vs), 1226 (w), 1168 (w), 946 (w), 810 (s), 790 (s), 766 (w), 722 (w), 711 (w), 692 (w). EA: obs. C, 81.58%; H, 619%; N, 3.60%, calcd. C, 81.86%; H, 6.08%; N, 3.67% for C26H23NO2.d) of 26  was isol2-(4′-tert-Butyl-biphenyl-4-ylmethyl)-3-methyl-naphthoquinone (27). tert-Butylphenylboronic acid was used as the boronic acid coupling partner for compound 25. After chromatography on silica gel (dichloromethane:petroleum ether/3:1), 113 mg  of 27 : δ = 8.06–8.13 , 7.66–7.71 , 7.41–7.50 , 7.24–7.30 , 4.06 , 2.28 , 1.35 . 13C-NMR : δ = 185.34 (Cq), 184.63 (Cq), 150.13 (Cq), 145.25 (Cq), 144.40 (Cq), 139.24 (Cq), 137.85 (Cq), 136.75 (Cq), 133.45 (CH), 132.12 (Cq), 132.03 (Cq), 128.93 (CH), 127.21 (CH), 126.59 (CH), 126.46 (CH), 126.25 (CH), 125.64 (CH), 34.47 (Cq), 32.08 (CH2), 31.32 (CH3), 13.30 (CH3). EI MS ): 394.3 , 379.2 (100). IR: 3439 cm−1 , 2962 (w), 1659 (vs), 1620 (m), 1595 (m), 1498 (w), 1462 (w), 1377 (w), 1334 (w), 1295 (s), 1182 (w), 1114 (w), 976 (w), 815 (m), 790 (w), 713 (w), 568 (w). EA: obs. C, 85.16%; H, 6.66%, calcd. C, 85.25%; H, 6.64% for C28H26O2.d) of 27  was isol2-(4′-nitro-Butyl-biphenyl-4-ylmethyl)-3-methyl-naphthoquinone (28). 4-Nitrophenylboronic acid was used as the boronic acid coupling partner for compound 25. After chromatography on silica gel (dichloromethane:petroleum ether/3:1), 67 mg  of 28 : δ = 8.23–8.26 , 8.05–8.11 , 7.65–7.73 , 7.51 , 7.34 , 4.08 , 2.27 . 13C-NMR : δ = 185.27 (Cq), 184.65 (Cq), 147.18 (Cq), 147.00 (Cq), 144.86 (Cq), 144.68 (Cq), 139.16 (Cq), 136.95 (Cq), 133.63 (CH), 132.12 (Cq), 131.98 (Cq), 129.40 (CH), 127.65 (CH), 127.59 (CH), 126.52 (CH), 126.38 (CH), 124.12 (CH), 32.22 (CH2), 13.40 (CH3). EI MS ): 383.3 , 368.2 (100). IR (KBr): 3436 cm−1 , 3073 (w), 2934 (w), 1661 (vs), 1620 (m), 1596 (vs), 1513 (vs), 1485 (m), 1375 (w), 1344 (vs), 1295 (vs), 1261 (w), 1182 (w), 1111 (m), 974 (w), 852 (m), 821 (m), 787 (w), 745 (m), 711 (m), 693 (m), 555 (w). EA: obs. C, 72.95%; H, 4.47%; N, 3.56%, calcd. C, 72.79%; H, 4.68%; N, 3.54% for C24H17NO4 0.7 H2O.d) of 28  was isol50 values were determined using standard in vitro proliferation assays [The ICn assays . Erythro50) of intraerythrocytic parasite development, using the SYBRgreen I assay as described before [green staining of parasitic DNA [50 values were calculated using SigmaPlot.In vitro antiplasmodial activity is expressed as the 50% inhibitory concentration . The results are expressed as % reduction in cell growth/viability compared to untreated control wells and CC50 is determined. Compounds are tested at 5 concentrations (64–16–4–1–0.25 µM). When the CC50 is lower than 16 µM, the compound is classified as toxic.MRC-5ad libitum throughout the experiment. The weight of the individual animals did not differ too much from the group mean.Swiss mice  were allocated randomly to five groups of four animals each. Drinking water and food were available P. berghei (ANKA-strain) is maintained in the laboratory by weekly mechanical subpassage in Swiss mice. The infection inoculum was prepared by taking heparinized blood was collected from a clinically ill donor mouse (approximately 20% parasitaemia) and diluted in PBS to obtain an infection inoculum of 0.15 mL with about 4 × 108 infected erythrocytes. The infection-inoculum was given intraperitoneally. The untreated infected controls developed severe malaria with most animals showing severe clinical signs on day 4. Chloroquine was used as reference treatment (10 mg/kg ip for 5 days). Treatment with chloroquine resulted in 100% survival until day 7 with low parasitaemia at day 4 post infection. The tested compounds were evaluated after ip dosing at 50 mg/kg for 5 days.2- and O2-free argon (Sigma Oxiclear cartridge) before use. All the stock solutions were prepared in spectroscopic grade dimethylsulfoxide  by weighing solid products using a XA105 Dual Range (0.01/0.1 mg–41/120 g) balance Mettler Toledo  and the complete dissolution was achieved using an ultrasonic bath. The ionic strength was maintained at 0.1 M with sodium perchlorate , and all measurements were carried out at 25.0 ± 0.2 °C. CAUTION! Perchlorate salts combined with organic ligands are potentially explosive and should be handled in small quantities and with the adequate precautions [Distilled water was further purified by passing it through a mixed bed of ion-exchanger  and activated carbon (Bioblock Scientific ORC-83005) and was de-oxygenated by COcautions .7 and 17 were performed in water (0.1 M NaClO4) while those of compounds 3, 10, 11, 18 and 19 were performed in NaClO4 0.1 M/DMSO (91:9 v/v) for solubility reasons. Absorption spectrophotometric titrations versus pH titrations of plasmodione derivatives 3 (1.81 × 10−5 M), 7 (2.00 × 10−5 M), 10 (1.81 × 10−5 M), 11 (2.18 × 10−5 M), 17 (2.00 × 10−5 M), 18 (1.84 × 10−5 M) and 19 (1.79 × 10−5 M) were carried out by diluting stock solutions freshly prepared in spectroscopic grade DMSO in 40 mL of the corresponding solvent contained in a jacketed cell  maintained at 25.0 ± 0.2 °C . The free hydrogen ion concentration was measured with a combined glass electrode  filled with 0.1 M NaCl in water and an automatic titrator system 794 Basic Titrino (Metrohm) connected to a microcomputer (Tiamo light 1.2 program for the acquisition of the potentiometric data). The combined glass electrode was calibrated as a hydrogen concentration probe by titrating known amounts of hydrochloric acid  with CO2-free sodium hydroxide solution  [4 and NaOH solutions were freshly prepared just before use and titrated with sodium tetraborate decahydrate  and potassium hydrogen phthalate , respectively, using methyl orange (RAL) and phenolphthalein  as the indicators. The GLEE program [0/mV and slope of the electrode/mV pH−1) and to check carbonate levels of the NaOH solutions used (<5%). The cell was thermostated at 25.0 ± 0.2 °C by the flow of a Lauda E200 thermostat. A stream of argon, pre-saturated with water vapour, was passed over the surface of the solution. The initial pH was adjusted to ~2–3 with HClO4  and the absorption spectrophotometric titrations of the plasmodione derivatives versus pH (~3 < pH < ~9–10) were carried out by addition of known volumes of NaOH solutions using the automatic titrator of the 794 Basic Titrino device (DET method). After each addition, an absorption spectra was automatically and repeatedly recorded using a CARY 50 spectrophotometer  fitted with Hellma optical fibers  and an immersion probe made of quartz suprazil  and interfaced (Cetrib) with the potentiometric unit. The distribution curves of the protonated species of the plasmodione derivatives as a function of pH were calculated using the Hyss program [The acido-basic properties of plasmodione derivatives ibution) . The HCl program  was appl program .The spectrophotometric data were analyzed with the Specfit program ,65,66 whN(Me)2 or -N(Et)2 in the different position of the benzylic chain were prepared in a 4-step synthesis for antimalarial drug evaluation using the multiresistant P. falciparum Dd2 strain. Interestingly, the non-protonable 4′-NHBoc-amino-3-benzylmenadione was selected for its potent in vitro activity, suggesting that cytosol-food vacuole shuttling of redox-cyclers may not need to accumulate in the food vacuole through lysosomotropic effects. This compound also revealed to be active in vivo in P. berghei-infected mice. These observations encourage pursuing further optimization of 3-benzyl-menadiones to improve the pharmacokinetic profile of plasmodione that has been shown to fulfill the criteria required for early antimalarial lead compounds.In summary, new synthetic procedures were successfully developed for the preparation of biaryl analogues of the antimalarial plasmodione in a 2-step sequence based on Kochi-Anderson and Suzuki-Miyaura coupling reactions. 3-Benzylmenadione derivatives bearing a terminal -


The final wR2 (F2) values were 0.1399 [I > 2σ(I)]. Flack parameter = 0.00 (12).Crystal X-ray diffraction data were collected on a Aglient Technologies Gemini A Ultra system with Cu KR,6S,8R,10R,19R,20R)-3α,6β,29-trihydroxylupan-28-oic acid (1), -3α,29-dihydroxy-6-oxo-lupan-28-oic acid (2) and  -3α,6β,7β,29-tetrahydroxylupan-28-oic acid (3), are revised from the previously reported ursane-type structure to lupane-type triterpenoids.Finally, the structures of (3


S-glycosides and N-glycosides incorporating 1,2,3,4-tetrahydronaphthalene and or 1,2-dihydropyridines moieties has been described. The aglycons 2, 4, and 7 were coupled with different activated halosugars in the presence of basic and acidic medium. The preliminary in-vitro cytotoxic evaluation revealed that compounds 3c, 3f, 5c and 7b show promising activity. A molecular docking study was performed against tyrosine kinase (TK) (PDB code: 1t46) by Autodock Vina. The docking output was analyzed and some compounds have shown hydrogen bond (H-B) formation with reasonable distances ranged from 2.06 A° to 3.06 A° with Thr 670 and Cys 673 residues found in the specified pocket. No hydrogen bond was observed with either Glu 640 nor Asp 810 residues, as was expected from pdbsum.A facile, convenient and high yielding synthesis of novel  Tetrahydronaphthalene derivatives have attracted significant attention in the field of drug discovery because of their wide array of pharmacological activities, including action as non-steroidal glucocorticoid receptor modulators, anticancer, analgesic, anti-inflammatory, antimicrobial, antiplatelet aggregation, hypotensive, antiarrhythmic and anti-HIV effects ,9,10,11.On the other hand recent advances have implicated the role of tyrosine kinases (TK) in the physiology of cancer. Though their activity is tightly regulated in normal cells, they may acquire transforming functions due to mutation(s), over expression leading to malignancy ,19. The S-glycosidic or N-glycosidic bond formation. A molecular docking study was performed in order to interpret the activity expressed by compounds 3c, 3f, 5b, 5c, 7a and 7b.Regarding the above mentioned findings and our previous reports  and 24]24], the S-glycosides and N-glycosides was designed in such a way that they incorporate a tetrahydronaphthalene derivative attached to a glycosyl moiety through a S-linkage or N-linkage. Synthesis of our desired compounds was achieved by allowing the acetyl derivative 1-ethanone (1) to react directly with the appropriate aromatic aldehyde and cyanothioacetamide in the presence of ammonium acetate, to afford the corresponding 2-thioxo(1H)pyridines 2a–c, respectively  pyranosyl bromide was achieved in a basic medium to give the corresponding thioglycoside derivatives 3a–f in a good yields, respectively .A series of ectively , Table 1ectively , Table 14a,b was accomplished by heating the acetyl derivative 1 with malononitrile and the appropriate aldehyde in the presence of ammonium acetate .Also, one-step synthesis of 2-imino (1H)pyridines  acetate , Table 1te  crystal structure (PBD code = 1t46) was downloaded . In ordeThe affinity of any small molecule can be considered to be a unique tool in the field of drug design, as there is a relationship between the affinity of organic molecules and the free binding energy and this can contribute in prediction and interpretation of the activity of organic compounds toward a specific target protein. Here we used the Autodock Vina ,26 progrThe most visible feature that was observed in the docking output that all compounds fitted well inside the same docking site and hence, the next step was to compare their affinities and try to rank them as observed in the previous table and then to measure the ligands’ interaction to find out which ones interact well and correlate that to the activity.7b, 3f, 5c, 5b, 3c and 7a, respectively . Compounds 3c and 7b were found to have low RMSD values that reflect their better fit as they were expected to have a conformation with a low deviation from the expected best binding mode. No clear correlation between the biological activity and the calculated affinity was observed. The only observation that was found that the active compounds have the best affinities regardless of their ranking in activity.The six compounds with the highest calculated affinities were found to be compounds 3c THR 670  whether they were involved in these interactions or not. For compound  THR 670  was the 3f shows two hydrogen bonds with Thr 670 and Cys 673 , as shown in Compound  Cys 673 . Compoun2b, 3a–f, 5b, 5c, 7a and 7b were tested for in vitro cytotoxic activity at two different concentrations. The results  cells were used for cytotoxic activity screening. The selected synthesized analogues  results  are expr−1 scale using KBr discs. 1H-NMR and 13C-NMR were recorded on a JEOL 300 MHz Spectrometer; chemical shifts are expressed in δ (ppm) with reference to TMS. The mass spectra were run at 70 ev with Packard 5890 Thewlett GC/MS Spectrometer, using the Electron Ionization (EI) technique. Elemental analyses  were carried out at the Microanalytical Data Center, Faculty of Science, Cairo, Egypt, and were in full agreement with the proposed structures within ±0.4% of the theoretical values. Thin layer chromatography was performed on precoated (0.75 mm) silica gel GF254 plates . Visualization was performed by illumination with UV light source (254 nm). Compounds 2b, 2c, 4a, 4b, 6a and 6b were prepared using the previously published procedures [Melting points  were determined in open glass capillaries using a Barnstead 9001 Electrothermal melting point apparatus. Infrared (IR) spectra were recorded on a Perkin Elmer FT-IR Spectrum BX Spectrometer at cmocedures  and 11]−1 scale 1 , 2-cyanothioacetamide , the an appropriate aromatic aldehyde, namely 2-furfural, p-nitrobenzaldehyde, and/or 3-ethoxy-4-hydroxybenzaldehyde (0.00172 mol) and excess ammonium acetate  was gently refluxed in absolute ethanol (15 mL) for 3–8 h. On cooling, the separated solid was filtered off, washed successively with water, and then with ether and recrystallized with aqueous ethanol to give 2a–c respectively -6--2-thioxo)-1,2-dihydropyridine-3-carbonitrile (2a). IR:  3,300 (NH), 2,200 (CN) and 1,206 (C-O). 1H-NMR (DMSO-d6): δ 1.76 , 2.78  attached to the aromatic ring of tetrahydronaphthalene), 6.89–8.61  and 13.9 (NH). 13C-NMR (DMSO-d6): δ 22.9 (-CH2-CH2-), 29.2 [2(CH2)] attached to the aromatic ring of tetrahydronaphthalene), 114.2 (CN), 106.2, 107.6, 117.7, 117.9, 125.66, 128.7, 129.3, 129.8, 137.8, 141.5, 142.1, 147.5, 148.3, 152.6, 180.4 (Ar-C), MS, m/z (%): .2a–c (0.01 mol) in ethanol (10 mL) and potassium hydroxide  in distilled water (6 mL) was added to a solution of 2,3,4,6-tetra-O-acetyl-α-D-gluco- or galactopyranosyl bromide  in acetone (30 mL). The reaction mixture was stirred at room temperature until completion (TLC) 20–24 h. and then the solution was evaporated and poured onto ice/cold water. The formed solid product was collected by filtration. The residue was washed with distilled water to remove the formed potassium bromide. The resulting solid product was dried and crystallized from dilute ethanol (A mixture of  ethanol .4-(Furan-2-yl)-6--2-pyridine-3-carbonitrile (3a). IR:  2937 , 2,216 (CN), 1,748 (C=O) and 1,230 (C-O). 1H-NMR (DMSO-d6): δ 1.78 , 2.02–2.03 , 2.88  attached to the aromatic ring of tetrahydronaphthalene), 4.02–4.06  4.31 , 5.03  5.26 , 5.71 , 6.24  and 6.84–8.13 . 13C-NMR (DMSO-d6): δ 22.9 (-CH2-CH2-), 29.2 [2(CH2)] attached to the aromatic ring of tetrahydronaphthalene), 115.7 (CN), 62.29 , 68.20 (C-4'), 69.76 (C-2'), 73.1 (C-3'), 75.17 (C-5'), 83.90 (C-1'), 106–180 (Ar-C) and 169.5–170.3 (4 × CO). MS, m/z (%): .4-(4-Nitrophenyl)-6--2-pyridine-3-carbonitrile (3b). IR:  2,930 , 2,217 (CN), 1,752 (C=O) and 1,348 & 1,528 (NO2). 1H-NMR (DMSO-d6): δ 1.75 , 2.04 , 2.76  attached to the aromatic ring of tetrahydronaphthalene), 4.04–4.06  4.35 , 5.05  5.24 , 5.74 , 6.31  and 7.1–8.4 . 13C-NMR (DMSO-d6): δ 22.18 (-CH2-CH2-), 19.62–19.90 (4 × -COCH3), 28.65 [2(CH2)] attached to the aromatic ring of tetrahydronaphthalene), 61.5 , 67.2 (C-4'), 69.5 (C-2'), 72.9 (C-3'), 74.6 (C-5'), 79.6 (C-1'), 115.46 (CN), 102–147 (Ar-C), 169.5 (4 × CO).MS, m/z (%): .4-(3-Ethoxy-4-hydroxyphenyl)-6--2-pyridine-3-carbonitrile (3c). IR:  3,458 (OH), 2,937 , 2,216 (CN), 1,754 (C=O) and 1,125 (C-O). 1H-NMR (DMSO-d6): δ 1.36 , 1.78 , 1.9–2.03 , 2.87  attached to the aromatic ring of tetrahydronaphthalene), 4.01–4.35 , 4.9  5.2 , 5.7 , 6.29  6.94–8.1  and 9.6 (OH). 13C-NMR (DMSO-d6): δ 14.49 (-OCH2CH3), 20.28 (4 × -COCH3), 22.49 (-CH2-CH2-), 28.63 [2(CH2)] attached to the aromatic ring of tetrahydronaphthalene), 61.54 , 63.89 (-O-CH2-CH3), 67.52 (C-4'), 69.50 (C-2'), 72.89 (C-3'), 74.63 (C-5'), 79.60 (C-1'), 115.75 (CN), 113–158 (Ar-C) and 169.45 (4 × CO). MS, m/z (%): .4-(Furan-2-yl)-6--2-pyridine-3-carbonitrile (3d). IR:  2,936 , 2,216 (CN), 1,754 (C=O) and 1,239 (C-O). 1H-NMR (DMSO-d6) δ 1.78 , 1.96–2.14 , 2.88  attached to the aromatic ring of tetrahydronaphthalene), 3.95 , 4.10 , 4.53  5.25 , 5.41 , 6.22 , 6.8–8.1 . 13C-NMR (DMSO-d6): δ 22.9 (-CH2-CH2-), 29.2 (2(CH2) attached to the aromatic ring of tetrahydronaphthalene), 115.7 (CN), 61.70 , 67.74 (C-4'), 69.45 (C-2'), 72.64 (C-3'), 74.97 (C-5'), 84.74 (C-1'), 106–180 (Ar-C) and 169.03–169.87 (4 × CO). MS, m/z (%): .4-(4-Nitrophenyl)-6--2-pyridine-3-carbonitrile (3e). IR:  2,932 , 2,217 (CN), 1751 (C=O) and 1,348 & 1,528 (NO2). 1H-NMR (DMSO-d6): δ 1.76 , 1.97–2.14 , 2.77  attached to the aromatic ring of tetrahydronaphthalene), 3.94 , 4.08 , 4.55  5.30 , 5.40 , 6.25  and 7.21–8.43 . 13C-NMR (DMSO-d6): δ 22.2 (-CH2-CH2-), 19.6–19.9 (4 × -COCH3), 28.82 (2(CH2) attached to the aromatic ring of tetrahydronaphthalene), 61.28 , 65.23 (C-4'), 66.67 (C-2'), 71.34 (C-3'), 74.46 (C-5'), 80.75 (C-1'), 115.46 (CN), 102–147 (Ar-C), 169.56–169.70 (4 × -CO of COCH3 groups). MS, m/z (%): .4-(3-Ethoxy-4-hydroxyphenyl)-6--2-pyridine-3-carbonitrile (3f). IR:  3,458 (OH), 2,217 (CN), 1,752 (C=O) and 1,056 (C-O). 1H-NMR (DMSO-d6): δ 1.39 , 1.80 , 1.96–2.06 , 2.79  attached to the aromatic ring of tetrahydronaphthalene), 4.08  4.11 , 4.13  4.5  5.26–5.65 , 6.25  and 6.98–8.08 . 13C-NMR (DMSO-d6): δ 13.96 (-OCH2CH3) 19.62–19.85 (4 × -COCH3), 22.17 (-CH2-CH2-), 28.81 [2(CH2)] attached to the aromatic ring of tetrahydronaphthalene), 61.25 , 64.06 (-O-CH2-CH3), 65.27 (C-4'), 66.73 (C-2'), 71.40(C-3'), 74.34 (C-5'), 80.69 (C-1'), 115.6 (CN), 110–147 (Ar-C) and 168.56–169.52 (4 × CO). MS, m/z (%): .4a and/or 4b (0.001 mol) in acetone (10 mL), 2,3,4,6-tetra-O-acetyl-α-D-gluco- or galactopyranosyl bromide  was added. A few drops of acetic acid were added and the reaction mixture was heated with stirring on a water bath at 40–50 °C for 25 h. The reaction mixture was cooled and then poured onto ice water. The formed precipitate was washed for several times with water and recrystallized from aqueous ethanol -6--2-pyridine-3-carbonitrile (5a). IR:  3,450 (NH), 2,990 (CH2), 2,216 (CN), 1,750 (C=O). 1H-NMR (DMSO-d6): δ 1.79 , 1.98–2.29  2.81 , 4.09 , 4.54 , 4.64 , 5.46 , 5.95 , 7.14–7.84 . 13C-NMR (DMSO-d6): 20.65–20.87 (4 × -COCH3), δ 22.2 (-CH2-CH2-), 28.7 (2(CH2) attached to the aromatic ring of tetrahydronaphthalene), 61.9 , 68.4 (C-4'), 68.8 (C-2'), 73.9 (C-3'), 75.25 (C-5'), 81.9 (C-1'), 115 (CN), 106–176 (Ar-C & pyridine-C), 169.5–170.3 (4 × CO). MS, m/z (%): .4--6--2-pyridine-3-carbonitrile (5b). IR:  3,360 (NH), 2,929 (CH2), 2,212 (CN), 1,737 (C=O). 1H-NMR (DMSO-d6): δ 1.74 , 2–2.01  2.76 , 4.01–4.06 , 4.51 , 4.71 , 5.47 , 5.62 , 6.53 , 7.14–7.84 . 13C-NMR (DMSO-d6): 20.69–19.89 (4 × -COCH3), 22.17 (-CH2-CH2-), 28.81 (2(CH2) attached to the aromatic ring of tetrahydronaphthalene), 61.94 , 66.22 (C-4'), 68.01 (C-2'), 71.73(C-3'), 74.33 (C-5'), 82.28 (C-1'), 115.6 (CN), 111–180 (Ar-C & pyridine C) and 169.56–170.33(4 × CO).MS, m/z (%): .4--6--2-pyridine-3-carbonitrile (5c).IR:  3,359 (NH), 2,932 (CH2), 2,214 (CN), 1,750 (C=O). 1H-NMR (DMSO-d6): δ 1.75 , 2–2.02  2.77 , 4.02 , 4.18 , 4.70 , 4.92 , 5.25 , 6.62 , 7.15–8.55 . MS, m/z (%):13C-NMR (DMSO-d6): 20.65–20.87 (4 × -COCH3), δ 22.4 (-CH2-CH2-), 28.91 (2(CH2) attached to the aromatic ring of tetrahydronaphthalene), 61.9 , 68.4 (C-4'), 68.8 (C-2'), 73.9 (C-3'), 75.25 (C-5'), 81.9 (C-1'), 115.8 (CN), 111.52–174.1 (Ar-C and pyridine-C), 169.5–170.3 (4 × CO). MS, m/z (%): .4--6--2-pyridine-3-carbonitrile (5d). IR:  3,430 (NH), 2,990 (CH2), 2,217 (CN), 1,750 (C=O). 1H-NMR (DMSO-d6): δ 1.75 , 1.99–2.01  2.76 , 4.05 , 4.2 , 4.68 , 4.95 , 5.37 , 6.65 , 7.15–9.54 . 13C-NMR (DMSO-d6): 20.69–19.89 (4 × -COCH3), 22.17 (-CH2-CH2-), 28.81 (2(CH2) attached to the aromatic ring of tetrahydronaphthalene), 61.94 , 66.22 (C-4'), 68.01 (C-2'), 71.73(C-3'), 74.33 (C-5'), 82.28 (C-1'), 115 (CN), 111–176 (Ar-C and pyridine C) and 169.56–170.33 (4 × CO). MS, m/z (%): .6a,b (0.01 mol) and conc. sulphuric acid (15 mL) was stirred at room temperature 6 h, and then poured on cold water. The formed solid product was collected by filtration and recrystallized from ethanol to give 7a,b -2-oxo)-1,2-dihydropyridine-3-carboxamide (7a). IR:  br. 3,382 (NH & NH2), 2,931 (CH2) and 1,668 (C=O). 1H-NMR (DMSO-d6): δ 1.69 , 2.70  attached to the aromatic ring of tetrahydronaphthalene), 4.03  and 6.40–8.89 . 13C-NMR (DMSO-d6): δ 22.97 (-CH2-CH2-), 29.23 [2(CH2)] attached to the aromatic ring of tetrahydronaphthalene), 107.17, 118.39, 124.78, 128.05, 128.42, 129.20, 129.56, 131.71, 137.98, 140.12, 140.68, 149.39, 153.11 (Ar-C), 163.88, 164.97 (2C=O). MS, m/z (%): .4--6--2-oxo-1,2-dihydropyridine-3-carboxamide (7b). IR:  br. 3,380 (NH & NH2), 2,932 (CH2) and 1,666 (C=O). 1H-NMR (DMSO-d6): δ 1.69 , 2.71  attached to the aromatic ring of tetrahydronaphthalene), 3.34 , 6.55–8.72  and 12.5 (NH). 13C-NMR (DMSO-d6): δ 22.98 (-CH2-CH2-), 29.18 (2(CH2) attached to the aromatic ring of tetrahydronaphthalene, 108.16, 111.59, 112.45, 118.37, 120.53, 123.07, 124.80, 128.41, 129.45, 130.15, 137.91, 140.51, 145.09, 148.75, 157.87, 159.81 (Ar-C), 163.20, 165.54 (2C=O). MS, m/z (%): .7a and/or 7b (0.001 mol) in acetone (10 mL), 2,3,4,6-tetra-O-acetyl-α-D-gluco- or galactopyranosyl bromide  was added. Few drops of acetic acid were added and the reaction mixture was heated with stirring on a water bath at 40–50 °C for 25 h. The reaction mixture was cooled and then poured onto ice water. The formed precipitate was washed for several times with water and recrystallized from ethanol -6--2-oxo)-1,2-dihydropyridine-3-carboxamide (8a). IR:  1H-NMR (DMSO-d6): δ 1.73 , 1.90–2.08 , 2.75  attached to the aromatic ring of tetrahydronaphthalene), 3.93  4.08 , 4.20  5.15–5.17 , 5.35 , 6.42–8.99  and 12.67 (NH). 13C-NMR (DMSO-d6): δ 19.85–20.04 (4 × -COCH3), 22.98 (-CH2-CH2-), 29.14 [2(CH2)] attached to the aromatic ring of tetrahydronaphthalene), 61.25 , 65.88 (C-4'), 67.34 (C-2'), 72.1 (C-3'), 74.34 (C-5'), 80.23 (C-1'), 107.17–153.09 (Ar-C) and 163.87–190.99 (6 × CO). MS, m/z (%): .4--6--2-oxo)-1,2-dihydropyridine-3-carboxamide (8b). IR: . 1H-NMR (DMSO-d6): δ 1.73 , 1.99–2.08 , 2.74  attached to the aromatic ring of tetrahydronaphthalene), 3.92  4.08 , 4.56  5.15–5.17 , 5.40 , 6.59–9.48  and 12.66 (NH). 13C-NMR (DMSO-d6): δ 19.85–20.04 (4 × -COCH3), 22.98 (-CH2-CH2-), 29.14 (2(CH2) attached to the aromatic ring of tetrahydronaphthalene), 61.25 , 65.88 (C-4'), 67.34 (C-2'), 72.1(C-3'), 74.34 (C-5'), 80.23 (C-1'), 108.17–159.79 (Ar-C) and 163.22–170.33 (6 × CO). MS, m/z (%): .Autodock vina allows the flexible docking of ligands into their site of action. It has the ability to use all the rotable bonds of the ligands to give a number of conformations from which the best mode could be deduced. The crystal structure of tyrosine kinase was downloaded from protein data bank . Pdb cod5 cells/mouse according to the method recommended by the Egyptian National Cancer Institute, Cairo University. Such developed tumor is characterized by its moderate rapid growth which could not kill the animal due to the accumulation of ascites before about 14 days after transplantation. Cells were counted before injection using the bright line haemocytometer and dilutions made by physiological saline. The desired number of cells was injected in a volume of 0.5 mL/mouse. Solid Ehrlich carcinoma was induced by inoculation of 2.5 × 105 cells in the back between the thighs of each animal [Ehrlich Ascites Carcinoma (EAC) cells was kindly provided by the National Cancer Institute . The tumor line was maintained in female Swiss albino mice by weekly intra-peritoneal injection of 2.5 × 10h animal .ad libitum.Adult female Swiss albino mice, obtained from the animal house of Cairo Cancer Institute  weighing 18–22 g were used. The local committee approved the design of the experiments, and the protocol conforms to the guidelines of the National Institutes of Health (NIH). The animals were maintained on a standard pellet diet and tap water 5 tumor cells per ml were suspended in phosphate buffer saline, then 25, 50, 100 µg/mL of the test compound were added to the suspension, kept at 37 °C for 2 h. Trypan blue dye exclusion test was then carried out to calculate the percentage of non-viable cells [A set of sterile test tubes were used, where 2.5 × 10le cells .O-acetyl-β-D-gluco/-galactopyranosyl) derivatives were synthesized and tested for their anticancer activity. The preliminary in vitro cytotoxicity evaluation revealed that five compounds show promised activity. This encouraged us to do a molecular docking study to find out a suitable binding mode for these compounds. The docking was done against TK and the docking output was analyzed. The compounds have shown hydrogen bond formation with a reasonable distance ranging from 2.06 A° to 3.06 A° with Thr 670 and Cys 673 residues. No hydrogen bond was observed with neither Glu 640 nor Asp 810 residues as was expected from pdbsum. From that we can conclude that substituted tetrahydronaphthalene  compounds may provide more activity by further study trying to modify their substitutions to give new derivatives with expected improved anticancer activity and that may be the subject of our future work.Some substituted tetrahydronaphthalene (2,3,4,6-tetra-


Alpinia galanga (Zingiberaceae), we newly isolated three new labdane-type diterpenes, termed galangalditerpenes A–C (1–3), along with four known sesquiterpenes (4–7) and two diterpenes (8 and 9). The stereostructures of 1–3 were elucidated on the basis of their spectroscopic properties. The melanogenesis inhibitory activities in theophylline-stimulated murine B16 melanoma 4A5 cells of these isolates, including the new diterpenes , were found to be more than 6–87-fold higher than that of arbutin (174 μM), a commercially available positive control.In our continuing study of biologically active natural products from the fruit of  Alpinia galanga Swartz, which is known as Greater Galangal in English and Kulanjan in Hindi, is widely cultivated in China, India, and Southeast Asian countries such as Thailand, Indonesia, and the Philippines 3). The positive-ion EIMS spectrum of 1 showed a molecular ion peak at m/z 318 (M+), and the molecular formula was determined as C20H30O3 by high-resolution MS. The UV spectrum exhibited maximum absorption at 230 nm, while the IR spectrum showed absorption bands at 1686 cm−1 ascribable to the conjugated aldehyde function. The 1H- and 13C-NMR spectra of 1 , heteronuclear multiple-quantum correlation (HMQC), and heteronuclear multiple bond correlation (HMBC) experiments ), six methylenes and a methylene bearing an oxygen function , 1.87 , H2-1), , 1.74 , H2-6), , 1.41 , H2-3), , 1.60 , H2-2), , 1.84 , H2-7), , 3.13 , H2-14), , 2.57 , H2-11), 3.62 and 4.35 ), two methines and an acetal methine , 1.64 , 5.44 ), a tri-substituted olefin ), and an aldehyde function ). As shown in 1H–1H COSY experiment on 1 indicated the presence of partial structures, shown by bold lines. In the HMBC experiment on 1, long-range correlations were observed between the following proton and carbon pairs: H-7 and C-8; H-9 and C-8, 10; H-12 and C-16; H-14 and C-12; H-16 and C-14; H2-17 and C-7–9, 15; H3-18 and C-3–5, 19; H3-19 and C-3–5, 18; H3-20 and C-1, 5, 9, 10. Thus, the linkage positions of the quaternary carbons  were clarified unambiguously, and the planar structure of 1 was elucidated. Next, the stereostructure of 1 was investigated using a nuclear Overhauser effect spectroscopy (NOESY) experiment, in which NOE correlations were observed between the following proton pairs: H-1α (δ 1.04) and H-5; H-2β (δ 1.51) and H3-19; H-3α (δ 1.19) and H-5; H-5 and H-7α (δ 1.64); H-6α (δ 1.15) and H3-19; H-7β (δ 1.84) and H-17β (δ 4.35); H-9 and H-17α (δ 3.62); H-12 and H-16; H-14α (δ 2.27) and H-15; H3-19 and H3-20, as shown in E)-8β,17-epoxylabd-15,16-dial, which was isolated from the seeds of A. galanga dec-4-en structure from natural sources.Galangalditerpene A  was obtained as a colorless oil with negative optical rotation ([α]3). In the positive-ion ESIMS profile, a sodium adduct ion peak was observed at m/z 341 [M + Na]+, and high-resolution ESIMS analysis revealed the molecular formula to be C20H30O3. The UV spectrum exhibited absorption maximum at 230 nm, while the IR spectrum showed absorption bands at 1728, 1712, and 1647 cm−1 ascribable to α,β-unsaturated lactone and aldehyde functions. The 1H- and 13C-NMR spectroscopic properties of 2 were generally superimposable onto those of galanolactone (8), but a difference was due to the 17α-formyl group in 2. As shown in 3), these were three methyls ), seven methylenes and a methylene bearing an oxygen function , 1.76 , H2-1), , 1.76 , H2-3), , 1.74 , H2-6), , 1.74 , H2-7), , 1.76 , H2-2), , 2.40 , H2-11), 2.70 and 2.74 , , 4.37 , H2-15)), three methines , 1.54 , 2.28 ), a tri-substituted olefin ), and an aldehyde function ). As shown in 2 was determined using 1H–1H COSY and HMBC experiments. NOE correlations in the NOESY experiment were observed between the following proton pairs, as shown in 3-20; H-3α (δ 1.20) and H-5; H-5 and H-7α (δ 1.40); H-6β (δ 1.35) and H3-20; H-7α (δ 1.40) and H-9; H-8 and H3-20; H3-19 and H3-20, and on this basis, the relative stereostructure of 2 was determined. To elucidate the absolute stereostructure, we carried out the conversion of 8 into 2. As shown in 8, the absolute configuration of which has been reported 3), and its molecular formula, C18H28O2, was determined using EIMS and high-resolution EIMS measurements. The 1H- and 13C-NMR spectra of 3 ), five methylenes and a methylene bearing an oxygen function , 1.42 , H2-1), , 1.43 (1H m), H2-3), 1.41 and 1.51 , , 1.97 , H2-7), , 1.71 , H2-6), and 2.31 and 2.38 ), two methines , 1.42 ), and a di-substituted trans-olefin , 6.49 ). The 1H and 13C-NMR spectroscopic properties of 3 were quite similar to those of (E)-15,16-bisnorlabda-8(17),11-diene-13-one (9), except for the signal due to the 17-position  in tetrahydrofuran (THF) yielded 3a  to give 11 fractions (Fr. 1 (1.16 g), Fr. 2 (2.91 g), Fr. 3 (2.95 g), Fr. 4 (6.45 g), Fr. 5 (9.80 g), Fr. 6 (11.47 g), Fr. 7 (1.25 g), Fr. 8 (10.03 g), Fr. 9 (2.80 g), Fr. 10 (16.28 g), and Fr. 11 (37.70 g)), as described previously . Fraction 3-3 (375.6 mg) was purified by HPLC ) to give (E)-15,16-bisnorlabda-8(17),11-diene-13-one . Fraction 4 (6.45 g) was subjected to reversed-phase silica gel CC  → MeOH) to afford eight fractions (Fr. 4-1 (62.4 mg), Fr. 4-2 (2.06 g), Fr. 4-3 (239.1 mg), Fr. 4-4 (53.2 mg), Fr. 4-5 (802.6 mg), Fr. 4-6 (252.8 mg), Fr. 4-7 (187.6 mg), and Fr. 4-8 (2.80 g)), as described previously [2O ] to give galangalditerpene C . Fraction 4-6 (252.8 mg) was purified by HPLC ) to give 1 . Fraction 5 (9.80 g) was subjected to reversed-phase silica gel CC  → MeOH) to afford nine fractions (Fr. 5-1 (63.8 mg), Fr. 5-2 (323.8 mg), Fr. 5-3 (2.57 g), Fr. 5-4 (483.9 mg), Fr. 5-5 (223.9 mg), Fr. 5-6 (2.08 g), Fr. 5-7 (827.2 mg), Fr. 5-8 (140.8 mg), and Fr. 5-9 (2.78 g)), as described previously , Fr. 9-4 (20.8 mg), Fr. 9-5 ), Fr. 9-6 (658.7 mg), and Fr. 9-7 (947.1 mg)).Dried fruit of Galangalditerpene A (1): A white powder; [α]c 0.35, CHCl3); UV ): 230 (3.42); IR (KBr) vmax cm−1: 1686, 1649, 1136, 1103; 1H- and 13C-NMR spectroscopic data, see m/z (%): 318 , 279 (9), 190 (100); HREIMS m/z 318.2197 .Galangalditerpene B (2): A colorless oil; [α]c 0.29, CHCl3); UV ): 230 (3.40); IR (KBr) vmax cm−1: 1728, 1712, 1647; 1H- and 13C-NMR spectroscopic data, see m/z: 341 [M + Na]+; HRESIMS m/z: 341.2104 [M + Na]+ .Galangalditerpene C (3): A colorless oil; [α]c 0.47, CHCl3); UV ): 230 (3.71); IR (film) vmax cm−1: 1697, 1635, 1123; 1H- and 13C-NMR spectroscopic data, see m/z (%): 276 , 258 (14), 43 (100); HREIMS m/z 276.2092 .8  in dry toluene (1.0 mL) at 0 °C was treated with BF3·Et2O , and the entire mixture was stirred at 0 °C for 30 min. The reaction mixture was poured into cold aqueous saturated NaHCO3 and extracted with EtOAc. The EtOAc extract was washed with brine, then dried over anhydrous Na2SO4 and filtered. Removal of the solvent under reduced pressure gave a crude product, which was purified by HPLC ) to furnish 2 .A solution of 3  in dry THF (1.0 mL) was treated with H2 in the presence of 5% Pd/C(en) (5.0 mg), and the entire mixture was stirred at room temperature for 6 h. The catalyst was removed by filtration, and the filtrate was evaporated under reduced pressure to give a crude product, which was purified by HPLC ) to furnish 3a .A solution of Dulbecco’s modified Eagle’s medium  was purchased from Sigma-Aldrich ; fetal bovine serum (FBS), penicillin, and streptomycin were purchased from Gibco ; and other chemicals used in this study were purchased from Wako Pure Chemical Co., Ltd. . The 48- and 96-well microplates (Sumilon) were purchased from Sumitomo Bakelite Co., Ltd. .2/air. Cells were harvested by incubation in phosphate-buffered saline (PBS) containing 0.05% ethylenediaminetetraacetic acid (EDTA) and 0.02% trypsin for ca. 5 min at 37 °C and used for the subsequent bioassays.Murine B16 melanoma 4A5 cells (RCB0557) were obtained from Riken Cell Bank . The cells were grown in DMEM  supplemented with 10% FBS, penicillin (100 units/mL), and streptomycin (100 µg/mL) at 37 °C in 5% COEffects on theophylline-stimulated melanogenesis and the viability of B16 melanoma 4A5 cells were determined as described previously ,27,28,29l-tyrosine or 3,4-dihydroxyphenyl-l-alanine (l-DOPA) as a substrate were determined as described previously [Tyrosinase activities using eviously ,27,28,29p) values less than 0.05 were considered significant.Values are expressed as mean ± S.E.M. One-way analysis of variance followed by Dunnett’s test was used for statistical analyses. Probability (1–3), were newly isolated from the 80% aqueous acetone extract of the fruit of A. galanga (Zingiberaceae) together with four known sesquiterpenes, clovane-2β,9α-diol (4), caryolane-1,9β-diol (5), (−)-2-oxoisodauc-5-en-12-al (6), and kobusone (7), and two diterpenes, galanolactone (8) and (E)-15,16-bisnorlabda-8(17),11-dien-13-one (9). The melanogenesis inhibitory activities in theophylline-stimulated murine B16 melanoma 4A5 cells of these isolates including the new diterpenes  were found to be more than 6–87-fold higher than that of arbutin (174 μM), a commercially used tyrosinase inhibitor. Considering the mechanisms of action, we examined the activity of the isolates against tyrosinase using either L-tyrosine or L-DOPA as a substrate. However, none of the isolates (1–9) showed inhibitory activity at the effective concentration. The detailed mechanisms of action of the melanogenesis inhibitory activity of the isolates require further examination.In conclusion, three new labdane-type diterpenes, galangalditerpenes A–C (


N-hydroxysuccinimide ester of O-acetylsalicylic acid. The procedure is based on a tandem C-acylation-cyclization process under mild reaction conditions. The structure of 3-methoxycarbonyl-4-hydroxy coumarin has been established by X-ray diffraction analysis and its geometry was compared with optimized parameters by means of DFT calculations.A novel short-step methodology for the synthesis in good yields of functionalized coumarins has been developed starting from an activated precursor, the  It is generally recognized that for an accurate description of hydrogen bonds at least double zeta quality basis augmented with a set of polarization and diffuse functions set is needed. Therefore a somewhat better geometry description is expected with 6–311++G standard basis set. All geometry optimization were followed by calculations of frequencies in order to identify obtained structures as energy minima (no imaginary frequencies). All minima for the three tautomeric forms were verified by establishing that the matrix of energy second derivatives (Hessian) has only positive eigenvalues . HOMO and LUMO frontier orbitals of the molecule were also computed at the same level of theory. All calculations were carried out with the Gaussian 09W program .DFT calculations have been performed for the tautomers of 3-methoxycarbonyl-4-hydroxy coumarin. Optimized molecular structures of the most stable tautomers are depicted in 4a which expected to be the predominant structure is shown in The optimized structure of compound The predominant tautomer a is coplanar. Major variation in geometry of different tautomeric forms are at C8-O3, C9-C10, C10-O4, C1-O1, O5-C11, C10-O5 bonds and hydrogen bonds at O1, O3 and O5 atoms . The remainder of the bonds and angles in the three tautomeric forms do not change significantly. Bond distances in the three tautomers indicate a more double bond character or a more single bond character as shown in Molecular orbital calculations provide a detailed description of orbitals including spatial characteristics, nodal patterns and individual atom contributions. The contour plots of the frontier orbitals for the ground state of 3-methoxycarbonyl-4-hydroxy coumarin are shown in The orbital energy levels of HOMO, second HOMO (HOMO-1), LUMO and second LUMO (LUMO+1) of 3-methoxy-4-hydroxy coumarin were deduced using the DFT/6–311++G method and are presented in 2Cl2 as the solvent. Results for relative energies and HOMO and LUMO orbitals are presented below in On top of our gas phase geometries we have also performed single-point C-PCM (Conductor like Polarized Continuum Model) calculations using CH2CO. Melting points were determined on a Gallenkamp MFB-595 melting point apparatus and are uncorrected. IR spectra were recorded on a Jasco 4200 FTIR spectrometer. NMR spectra were recorded on a Varian Gemini-2000 300 MHz spectrometer operating at 300 MHz (1H) and 75 MHz (13C). Chemical shifts δ are reported in ppm relative to DMSO-d6  and CDCl3 . J values are given in Hz. All reagents were purchased from Aldrich, Fluka and Acros and used without further purification. Dry THF was distilled from Na/PhO-acetylsalcylic acid  was treated under argon with N-hydroxysuccininimide  in anhydrous THF (11.5 mL), and a solution of DCC  in anhydrous THF (8.5 mL) was added dropwise at 0 °C. The reaction mixture was allowed to stir at 0 °C for 2 h. The resulting suspension was refrigerated overnight at 3-5 °C. The precipitated solid (DCCU) was filtered off and the filtrate was evaporated under reduced pressure and dried in vacuo to afford the N-hydroxysuccinimide ester of the corresponding O-acetylsalicylic acid as a white solid. Yield 2.4 g, 87%, mp 80 °C (lit. mp 87-89 °C [1H-NMR (DMSO-d6): δ 2.28 , 2.88 , 7.39-7.42 , 7.54 , 7.85 , 8.07-8.11 . 87-89 °C ). 1H-NMRN-hydroxysuccinimide ester of acetylsalicylic acid 2 (10 mmol) was added at 0 °C and the reaction mixture was stirred at r.t for 2h and then concentrated in vacuo. The obtained gum was diluted with H2O (10 mL) and washed with Et2O (10 mL). The aqueous extract was acidified with aqueous HCl (10%) in an ice-water bath to afford an oily product, which was extracted with CH2Cl2 (3 × 15 mL). The combined organic layers were dried over anhydrous Na2SO4, concentrated under reduced pressure and dried in vacuo to give the oily residue, which was treated either with method A or B.Sodium hydride  was added in anhydrous THF (65 mL) at 0 °C and the resulting mixture was stirred under argon for 15 min at room temperature. The appropriate active methylene compound  was then added at 0 °C, and the resulting mixture was stirred at room temperature for 1h. The C-acylation compound (10 mmol) was added to a solution of sodium  in absolute ethanol and stirred at room temperature for 24 h. The solvent was evaporated in vacuo and the residue was diluted with H2O and washed with Et2O, and the aqueous layer was acidified with 10% HCl at 0 °C to afford the desired coumarins as solid products. Method A: The 2Cl2 (3 × 15 mL). The combined organic layers were dried with Na2SO4, concentrated under reduced pressure and dried in vacuo to afford the desired coumarins as solid products.Method B: The C-acylation compound (10 mmol) was dissolved in MeOH (20 mL) and treated with aqueous HCl  for 48 h at room temperature to afford a gummy solid which was extracted with CH4-Hydroxy-3-methoxycarbonylcoumarin (4a): According to method A. White solid , mp: 139-140 °C : δ 4.02 , 7.30-7.38 , 7.65 , 8.00 , 14.55 ; 13C-NMR (CDCl3): δ 53.1 (COOCH3), 93.2 (C-3), 114.6 (C-4a), 117.1 (C-8), 124.4 (C-6), 125.0 (C-5), 135.8 (C-7), 154.5 (C-8a), 157.6 (C-2), 172.5 (C-4), 175.7 (COOCH3).9-140 °C ), IR (KB3-Ethoxycarbonyl-4-hydroxycoumarin (4b): According to method A. White solid , mp 98.5-100 °C : δ 1.43 , 4.50 , 7.28-7.36 , 7.67 , 8.00 , 14.73 ; 13C-NMR (CDCl3): δ 14.1 (COOCH2CH3), 62.9 (COOCH2CH3), 93.0 (C-3), 114.5 (C-4a), 116.8 (C-8), 124.2 (C-6), 125.0 (C-5), 135.5 (C-7), 154.2 (C-8a), 157.4 (C-2), 172.0 (C-4), 175.5 (COOCH2CH3).0-101 °C ), IR (KB3-Benzoyl-4-hydroxycoumarin (4c): According to method A. White solid , mp 115-116 °C : δ 7.30-7.70 , 8.10 , 16.72 ; 13C-NMR (CDCl3): δ 100.5 (C-3), 115.3 (C-4a), 117.4 (C-8), 124.6 (C-6), 125.7 (C-5), 128.1 (C-c), 128.5 (C-b), 132.7 (C-d), 136.3 (C-7), 137.8 (C-a), 155.2 (C-8a), 159.7 (C-2), 178.2 (C-4), 201.0 (COPh).6-117 °C ), IR (KB3-Cyano-4-hydroxycoumarin (4d): According to method B. White solid , mp 252-254 °C : δ 7.23 , 7.54 , 7.81 ; 13C-NMR (DMSO-d6): δ 75.0 (C-3), 116.3 (C-4a), 119.1 (C-8), 120.9 (CN), 123.0 (C-5), 124.9 (C-6), 132.6 (C-7), 153.5 (C-8a), 163.1 (C-2), 176.3 (C-4).0-251 °C ), IR /c, α = 3.802(3), b = 21.945(15), c = 11.352(8) A’, β= 90.097(10)°, V= 947,1(11) A’3, T=150(2) K, λ=0.71073 A’, Z = 4, 7326 reflections measured, 1686 unique (Rint = 0.0758), wR2 = 0.1897 , R1 = 0.0662 (I > 2σ(Ι). Data were collected on a Bruker APEX II diffractometer. The structure was solved by direct methods and refined on F2 using all the reflections [Compound lections . All theN-hydroxysuccinimide ester of acetylsalicylic acid as a new efficient precursor (scaffold). The structure of 3-methoxy-4-hydroxy coumarin has been determined by single-crystal X-ray diffraction and its geometry was compared with optimized parameters obtained by means of Density Functional Theory calculations at B3LYP/6-311++ level. A good agreement between theory and X-ray diffraction was found. The HOMO and LUMO levels and the lowest energy tautomer of 3-methoxy-4-hydroxy coumarin have been studied with DFT at B3LYP/6–311++G level. Further work in the benzopyranone series and the application of N-hydroxysuccinimide methodology towards the synthesis of more complex substrates with various substituents to explore potential biological applications will be reported in due course. In summary, we have successfully synthesized a range of functionalized 4-hydroxycoumarins using the


Xanthohumol (XN) and four minor hops prenylflavonoids: α,β-dihydroxanthohumol (2HXN), isoxanthohumol (IXN), 8-prenylnaringenin (8PN), and 6-prenylnaringenin (6PN), were tested for antiproliferative activity towards human cancer and normal cell lines. Nonprenylated naringenin (NG) was used as a model compound. Xanthohumol, α,β-dihydroxanthohumol and 6-prenylnaringenin were the most active compounds. Xanthohumol exhibited higher antiproliferative activity than cisplatin (CP) against five cancer cell lines: ovarian resistant to cisplatin A2780cis, breast MDA-MB-231 and T-47D, prostate PC-3, and colon HT-29. Isoxanthohumol was more potent than cisplatin against breast cancer cell lines MDA-MB-231 and T-47D whereas 6-prenylnaringenin was stronger than cisplatin against breast cancer cell line T-47D. It was found that tested chalcones possessed highly selective antiproliferative activity towards all tested breast cancer lines compared to the normal breast MCF 10A cell line . Low antiproliferative activity of naringenin indicates the importance of the prenyl group with respect to antiproliferative activity. Hop cones (hops) are a rich source of prenylated flavonoids. The major flavonoid found in hops with 0.1–1% of dry weight is xanthohumol (XN), the chalcone with a very broad spectrum of biological activities, such as antioxidant, anti-inflammatory and anticancer ones, suggesting a potential chemo-preventive effect ,3.Other prenylated flavonoids occurred at 10–100 fold lower concentrations than xanthohumol and, therefore, they were studied less intensely with the exception of 8-prenylnaringenin (8PN)—the most potent phytoestrogen so far known .Although xanthohumol is the major flavonoid found in hops, during wort boiling it undergoes thermal isomerisation to isoxanthohumol (IXN), which becomes the major hop flavonoid in beer (500 µg/L–3.44 mg/L)  or intesSo as to evaluate the effects of xanthohumol, isoxanthohumol and 8-prenylnaringenin on breast cancer tissue, Bolca and co-workers assessed their concentration, nature of metabolites, and biodistribution and compared them to 17β-estradiol . Phase IThe most common treatment for breast cancer patients involves using doxorubicin and docetaxel. However, therapeutic effects of these anticancer drugs are frequently accompanied by harmful side effects and also chemoresistance of cancer cells is often observed. Hence, there is a constant search for new effective and safe drugs . A part Delmulle and co-workers studied the antitumor activity of prenylated flavonoids against two prostate cancer cell lines: PC-3 and DU 145. In the case of PC-3 cells treated with isoxanthohumol and 6-prenylnaringenin and DU 145 cells treated with IXN, 8PN and 6PN, the formation of vacuoles was observed. The results showed that IXN, 6PN and 8PN, but not XN, induced a caspase-independent form of cell death and seem to be promising drugs for cancer therapy  for A-2780 to 187.10 ± 72.41 [µM] for MCF7), but its selectivity towards cancer cell lines compared to two normal cell lines (with the exception of PC-3 and MDA-MB-231) was within the range of selectivity index SI = 0.88–1.87. Naringenin  isoxanthohumol was more active than 8-prenylonaringenin. For the rest of the cell lines the cytotoxic activity of both compounds was similar. Comparing the sensitivity of all investigated cell lines to tested flavonoids one can observe that except for the hydrogenation of the α,β-double bond in xanthohumol, most of the structural modifications, such as isomerization of XN to IXN or demethylation of IXN to 8PN led to reduction of the cytostatic activity. Among the tested compounds, cisplatin exhibited higher antiproliferative activity only against three of the examined cancer cell lines: MCF7, A2780 and DU 145, and against one of the lines (HT-29), the activity of cisplatin was comparable to xanthohumol. However, this reference compound showed very high antiproliferative against normal human lung microvascular endothelial cells (HLMEC) and normal breast epithelial cell line (MCF 10A), which was 143 and 126-fold higher, respectively, than for tested flavonoids.50 for a normal cell line  to IC50 value for the respective cancerous cell line using the following equation:Commonly used anticancer drugs, such as cisplatin or doxorubicin have a big disadvantage of being toxic to both cancer and normal cells. Hence, to solve this problem, there is a search for new effective and safe pharmaceuticals. The selective action of a drug can be expressed by the selectivity index (SI), which is determined by comparing the cytotoxic activity of each compound to each cancerous cell line with that to a normal cell line. SI was calculated as a ratio of ICThe SI values higher than 1.0 indicate the compounds of considerable anticancer specificity, and SI much greater than 1.0—highly selective ones.A and SIB calculated for normal cell lines HLMEC and MCF 10A, respectively, are presented in SIBB = 5.16–9.91) and the highest one to the ovarian cancer cells (A2780) (SIB 1.94–2.85 times higher than cisplatin). The main prenylflavonoid present in human diet, isoxanthohumol, also showed very high selectivity to the A2780 line, and, what is more important, to cisplatin-resistant ovarian cancer cell line (A2780cis).All investigated compounds were selective towards the MCF 10A cell line  was noted for the chalcones XN and 2HXN against the A2780 cell line, which is almost 5-8 times higher than the reference. What is the most interesting is xanthohumol and α,β-dihydroxanthohumol were found to be selective agents towards all examined cancer cell lines, with the exception of MCF7 and PC-3 respectively (SIA).The selectivity of the tested compounds towards cancer cell lines in relation to the normal human lung microvascular endothelial cell line is not as high though. The highest SIThe results of our research indicate that all tested flavonoids, with particular emphasis on chalcones, are compounds with selective action, and as dietary supplements, they can effectively prevent carcinogenesis and work in conjunction with a known anticancer drug.Especially interesting are the results related to the breast cancer cells, because in developed countries they are the most often diagnosed tumors in women along with other estrogen-dependent cancers.Among the selected in this study of breast cancer and the mammary gland tumor cell lines, the most sensitive to the antiproliferative activity of flavonoids was the T-47D cell line, and the least sensitive were the cells of the MDA-MB-231 line. Our results need further verification in estrogen-sensitive tissue using a proper in vivo model, so as to find whether this activity depends on the estrogen receptor level in the cells.The examined compounds demonstrated high antiproliferative activity and very high selectivity with respect to cancer cells. They could be used as a dietary supplement, which can effectively prevent carcinogenesis in combination with a known anticancer drug.The isolation of xanthohumol from spent hops, the isomerization of XN into isoxantohumol, and chemical synthesis of 8-prenylnaringenin were described by Bartmańska et al. ,34. α,β-d6, CD3OD or acetone-d6. The structures of compounds  were con1H NMR (DMSO-d6) δH: 14.65 , 10.61 , 10.11 , 7.76 , 7,65 , 7.56 , 6.85 , 6.09 , 5.14 , 3.86 , 3.14 , 1.69 , 1.60 . 13C NMR δC: 191.7 (C=O), 164.7 (C-2′), 162.4 (C-4′), 160.6 (C-6′), 159.9 (C-4), 142.6 (C-β), 130.5 (C-3”), 129.9 , 126.1 (C-1), 123.8 (C-α), 123.0 (C-2”), 116.0 , 107.4 (C-3′), 104.6 (C-1′), 90.1, (C-5′), 55.8 (C-6′-OCH3), 25.5 (C-5”), 21.1 (C-1”), 17.7 (C-4”).Xanthohumol -4,2′,4′-trihydroxy-6′-methoxychalcone], 1H NMR (CD3OD): δH = 7.03 , 6.70 , 5.98 , 5.18 , 3.82 , 3.20 , 2.83 , 1.75 , 1.64 . 13C NMR δC: 206.1 (C=O), 165.7 (C-2′), 163.7 (C-4′), 162.6 (C-6′), 156.5 (C-4), 133.9 (C-1), 131.3 (C-3”), 130.3 , 124.2 (C-2”), 116.2 , 109.2 (C-3′), 105.8 (C-1′), 91.2 (C-5′), 55.9 (C-6′-OCH3), 47.5 (C-α), 31.6 (C-β), 26.0 (C-5”), 22.2 (C-1”), 17.9 (C-4”).α,β-Dihydroxanthohumol -4,2′,4′-trihydroxy-6′-methoxy-α,β-dihydrochalcone), 1H NMR (acetone-d6): δH = 10.43 , 9.53 , 7.28 , 6.77 , 6.14 , 5.32 , 5.09 , 3.70 , 3.11 , 2.92 , 2.57 , 1.58 , 1.53 ; 13C NMR δC: 188.8 (C=O), 162.0 (C-5), 161.9 (C-9), 160.1 (C-7), 157.8 (C-4′), 130.5 (C-1′), 130.1 (C-3”), 128.3 , 123.3 (C-2”), 115.5 , 107.9 (C-8), 105.0 (C-10), 93.1 (C-6), 78.3 (C-2), 55.8 (C-5-OCH3), 45.1 (C-3), 26.0 (C-5”), 22.0 (C-1”), 18.0 (C-4”).Isoxanthohumol -4,7-dihydroxy-5-methoxyflavanone), 1H NMR (DSMO-d6): δH = 12.10 , 10.80 , 9.60 , 7.31 , 6.78 , 5.96 , 5.41 , 5.08 , 3.19 , 3.07 , 2.72 , 1.58 , and 1.53 . 13C NMR: δC = 196.8 (C=O), 164.4 (C-7), 161.2 (C-5), 159.7 (C-9), 157.6 (C-4′), 130.2 (C-3″), 129.3 (C-1′), 128.1 , 122.70 (C-2″), 115.2 , 107.0 (C-8), 101.8 (C-10), 95.3 (C-6), 78.3 (C-2), 41.9 (C-3), 25.6 (C-5″), 21.3 (C-1″), and 17.6 (C-4″).8-Prenylnaringenin -5,7,4′-trihydroxyflavanone) 1H NMR (acetone-d6): δH = 12.46 , 9.74 , 8.64 , 7.40 , 6.89 , 6.03 , 5.41 , 5.23 , 3.23 , 3.15 , 2.72 , 1.75 , and 1.63 . 13C NMR δC = 196.40 (C=O), 163.7 (C-7), 161.3 (C-5), 161.0 (C-9), 157.7 (C-4′), 130.3 (C-3”), 129.9 (C-1′), 128.2 , 122.6 (C-2”), 115.2 , 108.0 (C-6), 102.1 (C-10), 94.3 (C-8), 78.9 (C-2), 42.6 (C-3), 24.90 (C-5”), 20.6 (C-1”), and 16.8 (C-4”). 6-Prenylnaringenin -5,7,4′-trihydroxyflavanone), 1H NMR and 13C NMR spectra of tested compounds can be found in The following established in vitro cell lines were applied: MCF7, T-47D, MDA-MB-231 (human breast cancer cell line), A2780 (human ovarian cancer cell line), A2780cis (human cisplatin-resistant ovarian cancer cell line), PC-3 and DU 145 (human prostate cancer cell lines), HT-29 (human colon cancer cell line), HLMEC , and MCF 10A .L-glutamine, 5% fetal bovine serum and 8 µg/mL of insulin (T-47D) or 1 mM sodium pyruvate (HT-29). MDA-MB-231, PC-3 and HLMEC cell lines were cultured in RPMI 1640 medium supplemented with 2 mM L-glutamine and 10% fetal bovine serum. Medium for MCF 10A cell line consisted of F-12 nutrient mixture supplemented with cholera toxin (0.05 µg/mL), insulin (10 µg/mL), hydrocortisone (0.5 µg/mL), 5% horse serum, and EGFH (20 ng/mL). All the culture media contained antibiotics: 100 units/mL penicillin , and 100 µg/mL streptomycin . All cell lines were grown at 37 °C with 5% CO2 humidified atmosphere.The cell lines: MDA-MB-231, HT-29, PC-3, DU 145, and MCF 10A were obtained from the American Type Culture Collection , MCF7, T-47D, A2780, A2780cis from the European Collection Cell Cultures , while the HLMEC cell line was obtained by M. Paprocka from the institute . All celTwenty-four hours before the incubation with the tested compounds, the cells were seeded in 96-plates  at a density of 1 × 104 cells per well (0.75 × 104 per well for MCF7 and 1 × 103 per well for HLMEC) in 100 µL of a culture medium.Each agent at each concentration (from 0.1 to 100 µg/mL) was tested in a single experiment, which was repeated four to five times. DMSO, which was used as a solvent (in a dilution corresponding to its highest concentration applied to tested compounds), did not exert any inhibitory effect on cell proliferation. Cisplatin was used as a reference drug.50 (inhibitory concentration 50%)—the concentration of the tested compound which inhibits 50% of the cells population. IC50 values were calculated for each experiment separately and mean values ± SD are presented in the The results were calculated as the ICAn assay was performed after 72 h of exposure to varying concentrations of the tested compounds (from 0.1 to 100 µg/mL); the time of incubation was chosen according to studies of Yong et al.  who analTested flavonoids have antiproliferative activity against human cancer cell lines and they are very selective.50 [µM]) and their selectivity indexes (SI) were much greater than one, i.e., T-47D, MDA-MB-231, and A2780cis.The anticancer activity is related to the structures of flavonoids. Comparing the antiproliferative activity and cancer selectivity of tested compounds, we came to the conclusion that there are chalcones that are promising potential chemotherapeutics, especially to these cancer lines, to which they showed higher antiproliferative activity than cisplatin (IC50 lower than for cisplatin) and more selective.The most promising compound concerning high antiproliferative activity against tested human cancer cell lines is xanthohumol, which is inexpensively accessible from waste (spent hops). However, in comparison to the activity on normal cell lines (selective action), it seems that chalcone 2HXN is even more interesting in the context of using it as a potent and selective anticancer drug. As far as the antiproliferative activity is concerned, the presence of a prenyl group is necessary, whereas O-methylation of the hydroxyl group at C-5 increased the cytotoxicity of the compounds against all tested cancer cell lines. The prenyl group at C-8 increased the antiproliferative activity, with the exception of the breast cancer cell lines. For these lines, especially T-47D, it was 6-prenylnaringenin that proved to be more active (ICThe results of our research provide valuable information, which after the complementary in vivo study may be useful for designing new medicines that are safe and devoid of harmful side effects.


The structures of all isolates were determined by spectroscopic analysis. The anti-inflammatory activity of the isolates was evaluated by testing their inhibitory effect on NO production in LPS-stimulated RAW 264.7 macrophages. Among the isolated compounds, 16-hydroxycleroda-3,13-dien-15,16-olide (6) and 16-oxocleroda-3,13-dien-15-oic acid (7) showed promising NO inhibitory activity at 10 µg/mL, with 81.1% and 86.3%, inhibition, respectively.Three new clerodane diterpenes, (4→2)- Polyalthia longifolia var. pendula, known as “Indian Mast Tree”, is a lofty evergreen tree which is distributed in tropical and subtropical regions. It is commonly cultivated in Asia and especially in Taiwan, as ornamental street tree due to its effectiveness in combating noise pollution  indicated that the α,β-unsaturated carboxylic acid is connected to C-9 by a short methylene chain [-C(11)H2-C(12)H2-] (trans- ring junction between rings A/B was suggested based on the NOESY cross peaks   with previous NMR data. We found close similarity of proton signals: δH 5.70 (H-14) and 2.15 (H-16); as well as carbon signals: δC 160.2 (C-13) and 115.8 (C-14) 169.4 (C-15) and 17.7 (C-16) with known solidagonal acid (4) and other similar structures  . Further(Heq-6)]  12,13].H2-] . Fu.H2-] . FH 5.70 H-/δC 169.4 169.4 C-, 34.5  signal in 2 was shifted downfield (δH 6.49) instead of δH 5.70 in 1. Also the disappearance of the methyl signal at δH 2.15 implied that the methyl group (H3-16) in 2 should be replaced by an aldehyde group (δH 9.55 (H-16)/δC 154.7 (C-13); δH 6.49 (H-14)/δC 194.4 (C-16), 154.7 (C-13)]. The C-2 position of the remaining aldehyde group was suggested by comparing the 1H-NMR spectra of 2 and 4 and was also supported by the HMBC correlations  , 9.55 (H-16] and carbons  are similar to compounds possessing similar fragments with trans-configuration such as 16-oxocleroda-3,13-dien-15-oic acid (7)  . An octaelations , which wgroup H3- in 2 shoacid (7) ,17 and 13 was predicted as C21H32O3 by the ESI-MS data, and it was thus deduced to have six IHD. In the 1H- and 13C-NMR spectra of 3, the downfield shifted proton signals  as well as two methine carbons  and two quaternary carbons  indicated the presence of an α,β-unsaturated lactone moiety, which was also supported by the UV absorption band at 207 nm and the IR absorption band at 1,758 cm−1. Moreover, a terminal double bond, one methoxy and three methyl groups were suggested based on certain characteristic proton signals  as well as the 13C-NMR signals of a methylene carbon [δC 103.0], a quaternary carbon [δC 159.0], a methoxy carbon [δC 56.1], and three methyl carbons . The detailed analysis of 1H and 13C-NMR data of 3 indicated that it possesses a skeleton similar to that of 16-hydroxycleroda-4(18),13-dien-15,16-olide (5)  confirmed that the junction of A/B ring in 3 is trans. The HMBC correlation [δH 3.43/δC 104.0] indicated that the additional methoxy is connected to C-16. Thus, the structure of 3 was assigned as 16(R&S)-methoxycleroda-4(18),13-dien-15,16-olide.The molecular formula of compound 4)  +2.0 , UV λmax (log ε) (MeOH) nm: 220 (3.24). IR νmax (KBr) cm−1: 3,361, 2,957, 2,922, 1,684, 1,654, 1,437, 1,208, 1,142. ESIMS (positive mode) m/z: 357.3 [M+Na]+. ESIMS (negative mode) m/z: 333.3 [M-H]−. HR-FAB-MS m/z: 335.2226 [M+H]+ . 1H-NMR  and 13C-NMR , see (4→2)-abeo-2,13-Diformyl-cleroda-2,13E-dien-14-oic acid (2). Oil. [α] +13.3 , UV λmax (log ε) (MeOH) nm: 219 (2.78). IR νmax (KBr) cm−1: 2,924, 2,853, 1,687, 1,676, 1,639, 1,564, 1,383, 1,208, 1,183. ESIMS (positive mode) m/z: 355.2 [M+Na]+. ESIMS (negative mode) m/z: 331.3 [M-H]−. HR-FAB-MS m/z: 355.1880 [M+Na]+ , 333.2060 [M+H]+ . 1H-NMR  and 13C-NMR , see 16(R&S)-Methoxycleroda-4(18),13-dien-15,16-olide (3). Oil. [α] −71.0 , UV λmax (log ε) (MeOH) nm: 207 (4.32). IR νmax (KBr) cm−1: 2,955, 2,924, 2,853, 1,758, 1,680, 1,639, 1,456, 1,206. ESIMS (positive mode) m/z: 355.24 [M+Na]+. ESIMS (negative mode) m/z: 331.20 [M-H]−. HR-FAB-MS m/z: 355.2251 [M+Na]+ , 333.2427 [M+H]+ . 1H-NMR  and 13C-NMR , see l-glutamine , and incubated in a 5% CO2 incubator at 37 °C [Murine RAW 264.7 macrophages were obtained from the American Type Culture Collection . Cells were propagated in RPMI-1640 medium supplemented with 10% heated-inactivated FCS and 2 mM at 37 °C .5 cells/mL, and then incubated with or without LPS (1 µg/mL) in the absence or presence of the tested compounds  for 24 h. The effect of the tested compounds on NO production was measured indirectly by determining the nitrite levels using the Griess reaction [RAW 264.7 cells were seeded in 24-well plate at a density of 5 × 10reaction .p values < 0.05 were considered to be significant. All tested compounds were re-purified by reversed-phase HPLC before the bioassay test (purity > 99%). All results are expressed as the means ± S.E. from three independent experiments. Data analysis involved one-way ANOVA with subsequent Scheffé test; P. longifolia var. pendula resulted in the isolation of three new clerodane diterpenes: (4→2)-abeo-cleroda-2,13E-dien-2,14-dioic acid (1), (4→2)-abeo-2,13-diformyl-cleroda-2,13E-dien-14-oic acid (2), and 16(R&S)-methoxycleroda-4(18),13-dien-15,16-olide (3), together with five known compounds 4–8. The inhibitory activity of the isolated compounds on NO production in LPS-stimulated macrophage RAW 264.7 was evaluated utilizing the Griess reaction. Among the tested compounds, 16-hydroxycleroda-3,13-dien-15,16-olide (6) and 16-oxocleroda-3,13(14)E-diene-15-oic acid (7) reduced NO production in LPS-induced cells in a dose-dependent manner with 81.1% and 86.3%, inhibition, respectively. These results add weight to the accumulating evidence supporting the potential application of P. longifolia var. pendula as a potent anti-inflammatory remedy.Chemical investigation of the unripe fruits of


Ephedra sinica, an important plant in Chinese traditional medicine, contains a complex mixture of proanthocyanidin oligomers as major constituents; however, only the minor components have been chemically characterized. In this study, oligomers with relatively large molecular weights, which form the main body of the proanthocyanidin fractions, were separated by adsorption and size-exclusion chromatography. Acid-catalyzed degradation in the presence of mercaptoethanol or phloroglucinol led to the isolation of 18 fragments, the structures of which were elucidated from their experimental and TDDFT-calculated ECD spectra. The results indicated that (−)-epigallocatechin was the main extension unit, while catechin, the A-type epigallocatechin–gallocatechin dimer, and the A-type epigallocatechin homodimer, were identified as the terminal units. Among the degradation products, thioethers of gallocatechin with 3,4-cis configurations, a B-type prodelphinidin dimer, a prodelphinidin trimer with both A- and B-type linkages, and a prodelphinidin dimer with an α-substituted A-type linkage were new compounds. In addition, a phloroglucinol adduct of an A-type prodelphinidin dimer, a doubly-linked phloroglucinol adduct of epigallocatechin, and a unique product with a flavan-3-ol skeleton generated by the rearrangement of the aromatic rings were also isolated. Ephedra sinica Stapf (Fam. Ephedraceae) is one of the most important plants in traditional medicine, and is used as a diuretic, antipyretic, diaphoretic, and for relieving a cough and asthma + peak at m/z 687.1382 in HRFABMS, indicating that 1 was a mercaptoethanol adduct of a prodelphinidin dimer with a B-type linkage. This was confirmed by the appearance of two intense signals at δH 6.57 and δH 6.67 (each 2H) arising from two pyrogallol-type B-rings and two methine proton signals attributable to C-ring H-2 in the 1H-NMR signals (H 4.34 (J = 9.4 Hz) was correlated to a carbon signal at δC 83.2, while the other F-ring H-2 (H 5.31 (br s) was found to be connected to the carbon that resonated at δC 75.05. The former indicated a 2,3-trans configuration and the latter, a 2,3-cis configuration .lleagues . Accordi2 showed the [M + H]+ peak at m/z 383.0801 in HRFABMS, confirming the molecular formula as C17H18O8S. The 1H-NMR spectrum (H 4.77 (J = 9.6 Hz), indicating the 2,3-trans configuration characteristic of gallocatechin. The C-ring H-4 resonated as a doublet at δH 4.36 (J = 4.3 Hz), which indicated the 3,4-cis configuration + peak at m/z 1011.1639 in HRFABMS. This implied that 3 was a thioether of a prodelphinidin trimer involving both A-type and B-type linkages. In the 1H-NMR spectrum  indicated the presence of three pyrogallol-type B-rings. The presence of two B-type linkages was apparent from the two C-ring H-2 signals resonating at δH 5.31 and δH 5.29 with small J2,3 values (<2 Hz). This again indicated that the two units were epigallocatechin. These spectroscopic features suggested a close relationship between 3 and the epigallocatechin trimer isolated from E. sinica with A- and B-type linkages +) confirmed the molecular formula as C21H18O10, the same as that of 11 and 12. Because of overlapping C-ring proton signals in the 1H-NMR spectrum measured in acetone-d6, the 2D NMR spectra were measured in methanol-d4  and phloroglucinol  rings, as well as mutually meta-coupled A-ring H-6 and H-8 , which were related to those observed in the spectra of 11 and 12 + peak at m/z 429.0818 in HRFABMS, indicating the molecular formula C21H16O10. An unambiguous assignment of the 1H- and 13C-NMR signals was achieved by 1H-1H COSY, HSQC, HMBC, and NOESY spectroscopy. The absence of a C-2 proton signal and appearance of a C-2 carbon signal at δC 100.21 confirmed the presence of an A-type linkage + ; 1H- and 13C-NMR data, see (+)-Gallocatechin-4-(2-hydroxyethyl)thioether (2): pale brown amorphous powder; c = 0.13, MeOH); UV (MeOH) λmax (log ε) 273 (3.17), 235 (4.42), 209 (4.72) nm; CD (MeOH) Δε220 −2.8, Δε249 +2.2, Δε284 −0.4; IR νmax 3344, 1621, 1537, 1515, 1455, 1345 cm−1; HRFABMS m/z 383.0801 [M + H]+ ; 1H- and 13C-NMR data, see (−)-Epigallocatechin--(−)-epigallocatechin-(4α→8)-(−)-epigallocatechin-4-(2-hydroxyethyl) thioether (3): reddish brown amorphous powder; c = 0.11, MeOH); UV (MeOH) λmax (log ε) 270 (3.83), 241 (4.65), 205 (5.31) nm; CD (MeOH) Δε218 −25.0, Δε233 +14.2, Δε284 −4.3; IR νmax 3276, 1615, 1541, 1445, 1348 cm−1; HRFABMS m/z 1011.1639 [M + Na]+ ; 1H- and 13C-NMR data, see (+)-Epigallocatechin--(+)-gallocatechin-4-(2-hydroxyethyl)thioether (7): pale brown amorphous powder; c = 0.15, MeOH); UV (MeOH) λmax (log ε) 270 (3.55), 247 (4.32), 208 (5.00) nm; CD (MeOH) Δε228 −25.6, Δε252 +3.7, Δε284 −3.0; IR νmax 3389, 1612, 1537, 1502, 1447, 1333 cm−1; HRFABMS m/z 707.1043 [M + Na]+ ; 1H- and 13C-NMR data, see 2O containing increasing proportions of MeOH to give 10 fractions. Fr. 1-1-5 (810.7 mg) was separated by Diaion HP 20SS column chromatography (3 cm × 21 cm) with H2O-MeOH to yield 12 (84.2 mg), 16 (103.7 mg), and five subfractions. Fr. 1-1-5-3 (454.4 mg) was successively separated by a Sephadex LH-20 (H2O-MeOH) and a Chromatorex ODS (H2O-MeOH) to afford 11 (24.4 mg). From Fr. 1-1-5-5 (59.8 mg), 17 (6.8 mg) was isolated by Chromatorex ODS chromatography (H2O-MeOH). Purification of 1-1-6 (102.3 mg) by Diaion HP20SS column chromatography (3 cm × 26 cm) with 0–100% MeOH in H2O yielded 13 (10.7 mg). Fractionation of 1-1-7 (120 mg) on a Diaion HP20SS (2 cm × 17 cm) with H2O-MeOH gave 5 (9.3 mg) and nine subfractions. Purification of Fr. 1-1-7-2 (6.3 mg) and Fr. 1-1-7-7 (11.4 mg) by a Chromatorex ODS (H2O-MeOH) furnished 13 (1.2 mg) and 17 (3.2 mg), respectively. Fr. 1-1-8 (210.7 mg) was separated by Diaion HP20SS chromatography to yield 8 (16.3 mg) and five subfractions, and subfraction 1-1-8-1 (59.4 mg) was further subjected to purification using a Sephadex LH 20 (2 cm × 16 cm) with EtOH-H2O-acetone  and then a Chromatorex ODS (H2O-MeOH) to give 14 (39.6 mg). Purification of Fr. 1-1-9 (204.8 mg) by Diaion HP 20SS chromatography (2 × 17 cm) with H2O-MeOH resulted in the isolation of 18 (11.4 mg).Fr. 1-1 was subjected to phloroglucinolysis according to the method reported by Kennedy and Jones and Bautista-Ortin and colleagues ,36, with2--4--3,4-dihydro-2H-1-benzopyran-3,5,7-triol  (13): pale brown amorphous powder; c = 0.11, MeOH); UV (MeOH) λmax (log ε) 270 (3.50), 234 (4.40), 210 (4.85) nm; CD (MeOH) Δε227 +0.8, Δε267 +1.2; IR νmax 3333, 1614, 1522, 1462, 1328 cm−1; HRFABMS m/z 431.0979 [M + H]+ ; 1H- and 13C-NMR data, see (−)-Epigallocatechin--(−)-epigallocatechin-4-phloroglucinol (14): pale brown amorphous powder; c = 0.14, MeOH); UV (MeOH) λmax (log ε) 270 (3.62), 235 (4.39), 214 (4.84) nm; CD (MeOH) Δε232 +18.9, Δε270 +1.5, Δε283 −1.2; IR νmax 3297, 1625, 1476, 1347 cm−1; HRFABMS m/z 733.1406 [M + H]+ ; 1H- and 13C-NMR data, see (−)-Epigallocatechin--phloroglucinol (16): pale yellow amorphous powder; c = 0.14, MeOH); UV (MeOH) λmax (log ε) 270 (3.62), 235 (4.39), 214 (4.84) nm; CD (MeOH) Δε216 +4.2, Δε246 +1.2, Δε270 −1.6; IR νmax 3297, 1625, 1476, 1347 cm−1; HRFABMS m/z 429.0818 [M + H]+ ; 1H- and 13C-NMR data, see A conformational search was performed using the Monte Carlo method with the MMFF94 force field using Spartan’14 . The resulting low-energy conformers within 6 kcal/mol of the global minimum were optimized at the B3LYP-SCRF/6-31G level in MeOH (PCM). The vibrational frequencies were also calculated at the same level to confirm that they were true minima, and no imaginary frequencies were found. The energies, oscillator strengths, and rotational strengths of the low-energy conformers with Boltzmann populations greater than 1% were calculated using TDDFT at the CAM-B3LYP-SCRF/6-31G level in MeOH (PCM). The ECD spectra were simulated by overlapping Gaussian functions with a 0.3 eV exponential half-width, and red-shifted by 25 nm. The calculated data for each conformer were averaged according to the Boltzmann distribution theory at 298 K based on their relative Gibbs free energies. All DFT calculations were performed using Gaussian 09 .E. sinica are mainly composed of oligomers, and in this study, the oligomers were separated and chemically characterized for the first time. Acid-catalyzed degradation with mercaptoethanol and phloroglucinol afforded 18 products, among which seven were previously unreported compounds. Epigallocatechin was the major extension unit, and catechin and A-type prodelphinidin dimers were identified as terminal units. The new compounds were characterized by spectroscopic analyses, and the stereochemistry of the trimeric products was determined with the aid of TDDFT calculations of the ECD spectra. Since E. sinica is one of the most important crude drugs in East Asia, and proanthocyanidins are major constituents with a comparable abundance to alkaloids, our results provide important insights into the molecular basis of traditional medicine.The proanthocyanidins of


Herreria montevidensis Klotzsch ex Griseb. (Herreriaceae) are used in traditional medicine by several Amerindian groups of the Paraguayan Chaco. Little is known on the chemistry of the plant, despite its widespread use across the South American Chaco. From the ethyl acetate/methanol 1:1 extract of the roots, four new and one known homoisoflavanoid, two flavan derivatives, a stilbene, a new alkaloid, and three new and four known spirostane steroids were isolated. The corresponding structures were elucidated by spectroscopic and spectrometric means. The homoisoflavonoids of the plant are related to compounds isolated from the Dracaenaceae (formerly Agavaceae) sources of the Chinese crude drug Dragon’s Blood. The new alkaloid is a novel skeleton that can be used as a chemical marker of Herreria. The spirostane steroids suggest chemotaxonomic relations with the Liliales. This is the first comprehensive report on the chemistry of a South American Herreria species.The roots of the South American vine  Herreria montevidensis Klotzsch ex Griseb. (family Herreriaceae) is abundant in the Chaco domains of South America. It is used in traditional medicine by many Amerindian groups of the Paraguayan Chaco. The Ayoreo rubbed the stem sap on the knees of children to accelerate growth + (100), 178 [M − C6H4(OH)CH3]+ (40), 107 [C7H7O hydroxytropylium]+ (100); HR-EI-MS 286.1205 .(3R)-7-Methoxy-3-(4-hydroxybenzyl)chroman (2). Colorless resin; 1H-NMR and 13C-NMR see m/z (rel. int.): 270 [M]+ (100), 162 [M − C6H4(OH)CH3]+ (40), 107 [C7H7O hydroxytropylium]+ (100); HR-EI-MS 270.1256 ; c = 2 × 10−3 g/100 mL, MeOH).(3R)-5,7-Dimethoxy-6-methyl-3-(4-hydroxybenzyl)chroman (3). Colorless resin; 1H-NMR and 13C-NMR see m/z (rel. int.): 314 [M]+ (100), 206 [M − C6H4(OH)CH3]+ (35), 107 [C7H7O hydroxytropylium]+ (30); HR-EI-MS 314.1518 . c = 15 × 10−3 g/100 mL, MeOH).(3R)-7-Hydroxy-5-methoxy-6-methyl-3-(4-hydroxybenzyl)chroman (4). Colorless resin; 1H-NMR and 13C-NMR see m/z (rel. int.): 300 [M]+ (100), 192 [M − C6H4(OH)CH3]+ (30), 107 [C7H7O hydroxytropylium]+ (30); HR-EI-MS 300.1362 . c = 21 × 10−3 g/100 mL, MeOH).7-Hydroxy-8-methoxy-3-(4-hydroxybenzyl)-3-chromen (5). Pale yellow resin; 1H-NMR and 13C-NMR see m/z (rel. int.): 284 [M]+ (100), 177 [M − C6H4(OH)CH2]+ (100), 107 [C7H7O hydroxytropylium]+ (85); HR-EI-MS 284.1059 . c = 8 × 10−3 g/100 mL, MeOH).Herrerin (9). Yellow needles, m.p. 228 °C; 1H-NMR and 13C-NMR see max (KBr) 3307, 2936, 1580, 1571, 1473, 1442, 1357, 1311, 1251, 1163, 1115, 1049, 963, 731, 673, 549, 433 cm−1; EI-MS m/z (rel. int.): 236 [M]+ (98), 207 (14), 143 [M − C8H7N]+ (100); HR-EI-MS 236.0950 .14α-Hydroxygitogenin ((25R)-5α-Spirostan-2α,3β,14α-triol) (11) isolated as 2O,3O-diacetate. White solid, 1H-NMR and 13C-NMR see max (KBr) 3564, 2929, 2859, 1741, 1450, 1368, 1252, 1180, 1134, 1098, 1043, 981, 902, 869, 734, 675, 609 cm−1; EI-MS m/z (rel. int.): 516 [M]+ (12), 457 [M − OAc]+ (6), 444 (20), 402 (18), 387 (28), 373 (32), 149 (56), 139 [C9H15O]+ (100); HR-EI-MS 516.3451 .14α-Hydroxyneotigogenin ((25S)-5α-Spirostan-3β,14α-diol) (14) isolated as 3O-acetate. White solid, 1H-NMR and 13C-NMR see m/z (rel. int.): 516 [M]+ (12), 457 [M − OAc]+ (6), 444 (20), 402 (18), 387 (28), 373 (32), 149 (56), 139 (C9H15O)+ (100); HR-EI-MS 516.3451 .14α-Hydroxyneogitogenin ((25S)-5α-Spirostan-2α,3β,14α-triol) (15) isolated as 2O,3O-diacetate. White solid, 1H-NMR and 13C-NMR see max (KBr) 3542, 2941, 1743, 1729, 1449, 1371, 1307, 1232, 1178, 1133, 1059, 1001, 945, 920, 900, 866, 735, 678, 649, 605 cm−1; EI-MS m/z (rel. int.): 532 [M]+ (12), 514 [M − H2O]+ (5), 473 [M − OAc]+ (10), 460 (20), 418 [460 − ketene]+ (15), 400 [460 − AcOH]+ (85), 385 [400 − Me]+ (8), 340 [400 − AcOH]+ (10), 167 (35), 149 (95), 139 [C9H15O]+ (100); HR-EI-MS 532.3400 .14α-Hydroxyyuccagenin ((25R)-Spirost-5-en-2α,3β,14α-triol) (16) isolated as 2O,3O-diacetate. White solid, 1H-NMR and 13C-NMR see max (KBr) 3550, 2951, 2872, 1738, 1455, 1369, 1239, 1180, 1101, 1053, 980, 919, 900, 869, 732, 648, 606 cm−1. EI-MS m/z (rel. int.): 530 [M]+ (1), 512 [M − H2O]+ (1), 452 [512 − AcOH]+ (4), 392 [452 − AcOH]+ (12), 377 [392 − Me]+ (45), 177 (68), 139 [C9H15O]+ (50), 43 (100); HR-EI-MS 530.3244 .2–5, a new alkaloid 9 belonging to a novel skeleton type and three new spirostane steroids 11, 15 and 16 were isolated and identified from the roots of H. montevidensis. The findings show clear chemotaxonomic relations with the Liliaceae and Agavaceae plant family, but with distinctive compounds that up to this point have not been identified in other species. To confirm the possible effect of the crude drug in traditional medicine, additional studies are needed, including bioactivity testing using suitable bioassays and chemical profiling of the extracts using hyphenated techniques.Several new compounds, including the rare homoisoflavanes


Their structures were elucidated on the basis of spectroscopic analyses and comparison with those previous literature data.A new cembrane diterpene, 6-acetoxy-7,8-epoxynephthenol acetate ( Nephthea  are a rich source of sesquiterpenoids, diterpenoids and steroids with diverse chemical structures and interesting biological activities , an acetoxyisopropyl residue , an oxymethine , a trisubstituted epoxide , two trisubstituted double bonds , and three tertiary methyls . Compound ral data  revealed13C-NMR spectra of 1 closely resembled those of 2 except for the presence of one additional acetoxy group. Therefore, it was suggested that compound 1 was a common 14-membered cyclic cembrane with an acetoxy, an acetoxyisopropyl and an epoxide functionalities. Assignments were performed based on 1H–1H COSY, HSQC and HMBC spectra data. 1H–1H COSY experiment revealed the sequences of the correlations depicted by the bold lines in In addition, the 1, the attachment of the acetoxyisopropyl group to C-1 was confirmed by correlations between H-1 to C-15, H-2 to C-15, H-16 to C-1 and H-17 to C-1. HMBC correlations between H-7 to C-8, H-9 to C-7 and C-8, H-10 to C-8 and H-19 to C-7, C-8 and C-9 were key for confirming that the methyl-bearing epoxide should be placed at the 7,8 positions. The vinyl methyl group at C-4 was confirmed by HMBC correlations between H-2 to C-4, H-5 to C-3 and C-4 and H-18 to C-3, C-4 and C-5. The other vinyl methyl group at C-12 was revealed by correlations between H-10 to C-12, H-11 to C-13, H-13 to C-12 and H-20 to C-11, C-12 and C-13. In addition, the chemical shift for C-6  clearly indicated that the acetoxy group was attached to the oxymethine carbon at C-6. Based on these findings, the gross structure of 1 was determined to be as shown in In the HMBC experiment of 1 was deduced from the NOESY experiments -6,15-diacetoxy-7,8-epoxycembra-3,11-diene. Compound 2 is reported to have R configuration at C-1 + ; 1H-NMR and 13C-NMR spectral data: see Colorless oil; [α]:25D –15.5 ; 13C-NMR (CDCl3) δ: 170.3 , 134.8 , 132.0 , 126.6 , 125.2 , 85.7 , 62.3 , 60.0 , 46.0 , 38.7 , 36.5 , 36.4 , 28.9 , 28.6 , 25.4 , 23.5 , 23.4 , 23.2 , 22.7 , 16.9 , 15.8 , 15.0 .Colorless oil; [α]:Escherichia coli (CSV01-08), Proteus mirabilis (CSV03-08), Pseudomonas aurelis (CSV04-08), Salmonella enteridis (CSV07-08), Salmonella thyphymunium (CSV08-08), Staphylococcus aereus (CSV09-08) and Vibrio cholerae (CSV10-08). The assay was performed as previously described [The antimicrobial bioassays for the isolated compounds were carried out using seven strains of human pathogenic bacteria: escribed .1) was isolated along with the known epoxynephthenol acetate (2), from a Nephthea sp. specimen collected from Layangan Island, Sabah. Their structures were established on the basis of spectral analysis. These findings have enriched our knowledge of the chemical constituents of Bornean soft corals. The isolated compounds 1 and 2 were tested for their antibacterial activities against human pathogenic bacteria by the disc diffusion method. Although both these compounds were found to have no activity, it is believed that they could pave the way to utilize secondary metabolites as chemotaxonomic markers for soft coral genus Nephthea.As a part of our chemical investigation on Bornean soft corals, a new cembrane diterpene, 6-acetoxy-7,8-epoxynephthenol acetate (