We previously demonstrated that uncoupling protein 1 activity as measured in isolated brown adipose tissue mitochondria (and as a native protein reconstituted into liposome membranes) was not activated by the non-flippable modified saturated fatty acid glucose-8. (CDCl3 126 174.3 (CH2 C-1) 63.05 (CH2 C-12) 51.39 (OCH3) 34.1 (CH2 C-2) 32.79 29.52 29.45 29.37 29.2 29.11 25.71 24.93 (each CH2); ES-HRMS calculated for C13H27O3 231.1960 found m/z 231.1917 [M?+?H]+. Methyl 12-(2 3 4 6 (20H ms aromatic H) 5.91 (1H t J 9.6?Hz H-3) 5.67 (1H t J 9.6?Hz H-4) 5.52 (1H dd J 9.6?Hz J 7.9?Hz H-2) 4.84 (1H d J 7.9?Hz H-1) Apixaban 4.64 (1H dd J 12.1?Hz J 3.4?Hz H-6a) 4.51 (1H dd J 12.1?Hz J 5.2?Hz H-6b) 4.16 (1H ddd J 9.6?Hz J 5.2?Hz J 3.4?Hz H-5) 3.91 (1H dt J 9.7 J` 6.3?Hz CHHO) 3.66 (3H s OCH3) 3.54 (1H dt J 9.7?Hz J 6.7?Hz CHHO) 2.29 (2H t J 7.6?Hz CH2CO2Me) 1 (18H m 9 13 NMR (126?MHz CDCl3) 174.24 166.1 165.8 165.17 165.02 (each C?=?O) 133.34 133.14 133.09 133.03 129.78 129.72 129.7 (each aromatic CH) 129.65 129.41 128.91 128.87 (each aromatic C) 128.34 128.33 128.29 128.27 128.23 (each aromatic CH) 101.28 (CH C-1) 72.97 (CH C-3) 72.16 (CH C-5) 71.95 (CH C-2) 70.3 (CH2O) 69.92 (CH C-4) 63.26 (CH C-6) 51.36 (OCH3) 34.08 29.38 29.37 29.34 29.18 29.11 26.89 25.74 24.92 (each CH2). ES-HRMS calculated for C47H52O12Na 831.3356 found 831.3391 [M?+?Na]+. Methyl 16-(2 3 4 6 (20H ms aromatic H) 5.9 (1H t J 9.7?Hz H-3) 5.66 (1H t J 9.7?Hz H-4) 5.51 (1H dd J 7.9?Hz J 9.7?Hz H-2) 4.83 (1H d J 7.9?Hz H-1) 4.63 (1H dd J 12.1?Hz J 3.3?Hz H-6a) 4.51 (1H dd J 12.1?Hz J 5.3?Hz H-6b) 4.15 (1H ddd J 9.7?Hz J 5.3?Hz J 3.3?Hz H-5) 3.91 (1H dt J 9.7?Hz J 6.3?Hz CHHO) 3.66 (3H s OCH3) 3.53 (1H dt J 9.6?Hz J 6.7?Hz CHHO) 2.3 (2H t J 7.6?Hz CH2CO2) Apixaban 1 (28H ms 14 13 NMR (126?MHz CDCl3) 174.31 166.14 165.84 165.21 165.06 (each C?=?O) 133.37 133.17 133.11 133.06 129.88 129.82 129.78 129.76 129.75 129.73 (each aromatic CH) 129.66 129.44 128.9 128.88 (each aromatic C) 128.43 128.38 128.38 128.33 128.3 128.27 (each aromatic CH) 101.31 (CH C-1) 72.99 (CH C-3) 72.18 (CH Rabbit Polyclonal to GSPT1. C-5) 71.97 (CH C-5) 70.37 (CH2O) 69.95 (CH C-4) 63.29 (CH2 C-6) 51.4 (OCH3) 34.13 29.64 29.62 29.59 29.53 29.46 29.42 29.26 29.16 25.79 24.97 (each CH2). ES-HRMS calculated for C51H60O12Na 887.3982 found 887.3981 [M?+?Na]+. 12 acid (1a) Protected glycoside 4a (72?mg 0.0891 was dissolved in MeOH (5?mL). A catalytic amount of NaOMe (1?M in MeOH) was added and the resulting solution was stirred for 2?h at room temperature. The reaction was quenched with Amberlite IR-120 (plus) until pH?=?6.0 the resin was filtered off and washed with THF-MeOH 1:1. The solvent was removed under reduced pressure and dried thoroughly under high vacuum. The white solid was taken up in H2O (2?mL) and THF was added until the solution became clear. To this LiOH?·?H2O (20?mg) was added and the Apixaban reaction mixture was stirred for 1?h. The reaction was quenched using Amberlite IR-120 (plus) until pH?=?6.0; the resin was filtered off and washed with THF-MeOH 1:1. The solvent was removed under reduced pressure. Apixaban Chromatography (1:9 MeOH-CH2Cl2) gave 1a (27?mg 81 as a white solid; IR (film) cm?1: 3 362 2 918 2 850 1 553 1 261 1 83 1 37 839 1 NMR (CD3OD 500 4.28 (1H d J 7.8?Hz H-1) 3.92 (1H dt J 9.5?Hz J 6.9?Hz CHHO) 3.89 (1H dd J 11.8?Hz J 2.0?Hz H-6a) 3.7 (1H dd J 11.8?Hz J 5.4?Hz H-6b) 3.57 (1H dt J 9.5?Hz J 6.9?Hz CHHO) 3.26 (3H overlapping signals H-3 H4 and H-5) 3.2 (1H dd J 9.1?Hz J 7.8?Hz H-2) 2.29 (2H t J 7.4?Hz CH2CO2H) 1.3 (18H ms 9 CH2); 13C NMR (CD3OD 125 174.62 (C?=?O detected indirectly using HMBC) 104.44 (CH C-1) 78.22 (CH C-3) 77.98 (CH C-4) 75.21 (CH C-2) 71.77 (CH C-5) 70.96 (CH2O) 62.87 (CH2 C-6) 30.86 30.75 30.68 30.64 30.53 30.5 27.15 (each CH2); ES-HRMS calculated for C18H33O8 377.2175 found 377.2159 [M???H]?. 16 acid (1b) The protected glycoside 4b (100?mg 0.0891 was dissolved in MeOH (5?mL). A catalytic amount of NaOMe (1?M in MeOH) was added and the resulting solution was stirred for 2?h at room temperature. The reaction was quenched with Amberlite IR-120 (plus) until pH?=?6.0; the resin was filtered off and washed with THF-MeOH 1:1. The solvent was removed under reduced pressure and the residue was dried thoroughly under high vacuum. The white solid was taken up in H2O (2?mL) and THF was added until the solution became clear. To this LiOH.