E was stirred for 24 h at space temperature. The reaction mixture was poured onto

E was stirred for 24 h at space temperature. The reaction mixture was poured onto ice and extracted with ethyl acetate (3 30 mL). The organic layer was dried more than anhydrous Na2 SO4 along with the solvent was evaporated below a vacuum. The crude reaction item was stirred in MeCN and hexane for 3 h and was then purified by column chromatography utilizing ethyl acetate/ethanol (10/1) mobile phase. The solvent was evaporated in vacuum to supply the solution (266 mg, 76 ) as a brown solid. 1 H NMR (500 MHz, DMSO-d6 , 25 C): = 11.40 (s, 1H, OH), 11.16 (s, 1H, OH), 7.35 (d, J = 1.9 Hz, 1H, ArH), 7.14 (d, J = 1.9 Hz, 1H, ArH), six.75 (s, 1H, ArH), 2.38 (s, 3H, CH3 ) ppm. 13 C NMR (126 MHz, DMSO-d6 , 25 C): = 158.two, 151.1, 146.8, 139.7, 134.7, 133.5, 126.7, 121.four, 112.1, 110.6, 109.4, 108.9, 105.0, 96.7, 14.6 ppm. IR: 3451, 1738, 1600, 1426, 1367, 1202, 1094 cm-1 . HRMS (ESI- ): m/z calcd for C15 H10 O8 S 349.0024 [M-H]- , identified: 349.0032 [M-H]- . 1,three,8-trimethoxy-6-methylanthracene-9,10-dione (E_OCH3 ) [30]. Potassium carbonate (415 mg, 3.0 mmol) was added to a resolution of emodin (one hundred mg, 0.37 mmol) in acetone (7 mL). Then dimethyl sulfate (285 , 3.0 mmol) was added gradually and the reaction mixture was stirred at -Irofulven MedChemExpress reflux for 24 h. The reaction mixture was allowed to cool to space temperature. Just after cooling to area temperature, the solvent was evaporated. Then water (5 mL) and acetone (five mL) were added for the reaction mixture under stirring for 15 min. The solution was filtrated off, washed with water, and dried in vacuum to supply the product (94 mg, 81 ) as a yellow hite solid. 1 H NMR (500 MHz, IL-4 Protein manufacturer Chloroform-d, 25 C): = 7.64 (s, 1H, ArH), 7.32 (s, 1H, ArH), 7.09 (s, 1H, ArH), 6.76 (s, 1H, ArH), three.98 (s, 3H, OCH3 ), three.96 (s, 3H, OCH3 ), three.95 (s, 3H, OCH3 ), two.47 (s, 3H, CH3 ) ppm. 13 C NMR (126 MHz, Chloroform-d, 25 C): = 184.four, 181.8, 163.7, 161.7, 159.8, 144.6, 136.4, 134.four, 121.5, 119.six, 119.0, 118.four, 105.three, 101.9, 56.five, 56.5, 55.9, 22.1 ppm. IR: 1657, 1599, 1322, 1241, 1021, 946, 910 cm-1 . HRMS (ESI ): m/z calcd for C18 H16 O5 313.1071 [MH] , identified: 313.1077 [MH] . 2,four,five,7-tetrabromo-1,three,8-trimethoxy-6-methylanthracene-9,10-dione (E_Br_OCH3 ). Potassium carbonate (415 mg, three.0 mmol) was added to a resolution of brominated emodin five (216 mg, 0.37 mmol) in acetone (7 mL). Then dimethyl sulfate (285 , three.0 mmol) was added slowly along with the reaction mixture was heated to reflux for 24 h. The reaction mixture was permitted to cool to space temperature. Following cooling to area temperature, the solvent was evaporated. Then water (5 mL) and acetone (5 mL) have been added towards the reaction mixture with stirring for 15 min. The solution was filtrated off, washed with water and dried inside a vacuum to provide the solution (202 mg, 87 ) as a light pink strong. 1 H NMR (500 MHz, Chloroform-d, 25 C): = four.02 (s, 3H, OCH3 ), four.01 (s, 3H, OCH3 ), three.98 (s, 3H, OCH3 ), two.78 (s, 3H, CH3 ) ppm. 13 C NMR (126 MHz, Chloroform-d, 25 C): = 184.1, 179.5, 160.1, 156.9, 155.1, 147.0, 135.eight, 135.0, 128.7, 128.0, 126.7, 121.7, 117.7, 112.2, 63.2, 63.1, 61.1, 25.9 ppm. IR: 2153, 2036, 1695, 1367, 1322, 1216, 991 cm-1 . HRMS: m/z calcd for C18 H12 Br4 O5 624.7491 [MH] , discovered: 624.7487 [MH] . 3.two. Evaluation of Antiviral Activity Compound preparation. For testing purposes, all compounds have been dissolved in DMSO to a final concentration of 50 mM. With these stock solutions, mother plates were prepared in DMSO and generated stocks for testing eight point ose responses. 1:1 dilutions had been prepared, startin.