H31O6: [M + H]+ 403.2115; located 403.2118. Synthesis of (3S,3aR,3a1R,6aR,7S,7aS,11aR,11bS)-7-hydroxy-5,five,eight,8-tetramethyl-15methylene-3,3a,7,7a,eight,11b-hexahydro-1H-6a,11a-(epoxymethano)-3,3a1ethanophenanthro[1,10-de][1,3]dioxine-9,14(2H)-dione (19) To a solution of 18 (10 mg, 0.025 mmol) in dichloromethane (two mL) was added PDC (11.2 mg, 0.03 mmol) at rt. The resulting mixture was stirred at rt for 4 h. The reaction mixture was then filtered, along with the filtrate was diluted with water and extracted with dichloromethane. The extract was washed with saturated NaHCO3 (aq.) option and brine, dried more than anhydrous Na2SO4, filtered, and evaporated to provide an oily residue. The residue was purified utilizing preparative TLC created by 50 EtOAc in hexane to afford the desired product 19 as a colorless amorphous gel (8.0 mg, 80 ). []25D -100 (c 0.ten, CH2Cl2); HPLC purity 97.five (tR = 18.62 min); 1H NMR (600 MHz, CDCl3) six.29 (d, 1H, J = 10.two Hz), 6.19 (s, 1H), 6.00 (d, 1H, J = 10.2 Hz), five.60 (s, 1H), five.54 (d, 1H, J = 12.0 Hz), 4.84 (s, 1H), 4.16 (d, 1H, J = ten.2 Hz), 4.07 (m, 1H), four.01 (d, 1H, J = ten.2 Hz), three.10 (d, 1H, J = eight.4 Hz), two.58 (m, 1H), 1.93 (d, 1H, J = 7.8 Hz), 1.88 (m, 2H), 1.76 (m, 1H), 1.66 (s, 3H), 1.60 (m, 2H), 1.37 (s, 3H), 1.36 (s, 3H), 1.27 (s, 3H); 13C NMR (150 MHz, CDCl3) 204.1, 203.two, 149.9, 142.six, 129.8, 121.two, 101.5, 95.four, 70.9, 69.8, 64.1, 56.2, 55.7, 48.three, 44.6, 40.1, 38.eight, 30.1, 29.9, 25.four, 23.470482-44-1 uses 9, 22.four, 17.1; HRMS Calcd for C23H29O6: [M + H]+ 401.1959; found 361.1962. Synthesis of (4aS,5S,6S,6aR,9S,11aS,11bR,14R)-5,6,14-trihydroxy-4,4-dimethyl-8methylene-4,4a,five,6,9,10,11,11a-octahydro-3H-6,11b-(epoxymethano)-6a,9methanocyclohepta[a]naphthalene-3,7(8H)-dione (20) To a remedy of 19 (15 mg, 0.037 mmol) inside a mixture of MeOH (2 mL) and CH2Cl2 (0.5 mL) was added five HCl aqueous option (0.five mL) at rt. The resulting mixture was stirred at rt for 4 h. The reaction mixture was then diluted with water and extracted with dichloromethane. The extract was washed with saturated NaHCO3 (aq.) resolution and brine, dried over anhydrous Na2SO4, filtered, and evaporated to give an oily residue. The residue was purified applying preparative TLC developed by 66 EtOAc in hexane to afford the preferred solution 20 as a colorless amorphous gel (11.5 mg, 85 ). []25D -128 (c 0.10, CH2Cl2); HPLC purity 98.two (tR = 14.87 min); 1H NMR (600 MHz, CDCl3) 6.31 (d, 1H, J = 10.2 Hz), six.22 (s, 1H), 6.13 (d, 1H, J = 11.four Hz), 6.02 (d, 1H, J = 10.eight Hz), five.63 (s, 1H), 4.92 (s, 1H), 4.17 (d, 1H, J = 10.2 Hz), 4.06 (dd, 1H, J = 1.8 Hz, ten.2 Hz), 3.98 (m, 1H), three.ten (d, 1H, J = 9.0 Hz), 2.58 (m, 1H), 1.95 (d, 1H, J = 9.0 Hz), 1.91 (m, 2H), 1.65 (m, 3H), 1.34 (s, 3H), 1.1831130-33-6 Chemscene 26 (s, 3H); 13C NMR (150 MHz, CDCl3) 206.PMID:24580853 0, 202.8, 150.five, 142.7, 130.0, 122.two, 97.7, 72.4, 72.1, 64.eight, 61.7, 55.6, 51.4, 44.four, 42.5, 39.2, 29.four, 23.6, 22.0, 17.five; HRMS Calcd for C20H25O6: [M + H]+ 361.1646; located 361.1651. In Vitro Determination of Effects of Synthesized Compounds on Cancer Cell Proliferation Breast cancer cell lines MCF-7 and MDA-MB-231 have been seeded in 96-well plates at a density of 1 ?104 cells/well and treated with DMSO, 0.01 M, 0.1 M, 1 M, 5 M, 10 M, and 100 M of person compound for 48 h, and after that 20 L of 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) (5 mg/mL in PBS) was added to every nicely and further incubated for a different four h. Then, MTT resolution was removedNIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptJ Med Chem. Author manuscript; avai.
