Trabecular bone parameters were assessed, including BV/TV, Tb

Trabecular bone parameters were assessed, including BV/TV, Tb.Th, Tb.N Fendiline hydrochloride and Tb.Sp. 2.2.4. MHz, CDCl3): 13.085. IR cm?1: 3427, 2981, 2927, 1714, 1537. HRMS (ESI) Fendiline hydrochloride (M + H) Calcd for C16H37N2O10P2: 479.1917. Found: 479.1923. 2.1.2. (((2-(8-((R)-3-methyl-1-((S)-3-phenyl-2-(pyrazine-2-carboxamido)propanamido)butyl)tetrahydro-8H-4I4,8I4-[1,3,2]oxazaborolo[2,3-b]-[1,3,2]oxazaborol-4-yl)ethoxy)carbonyl)amino)methylene)bis(phosphonic acid (BP-Btz) A dry 10 mL round bottom flask was charged with pre-activated (at 120 C) powdered 3? molecular sieves (1 g) and a solution of BP-Linker-Tetraester (125 mg, 0.26 mmol) and (120.5 mg of Bortezomib (Btz) (0.31 mmol, 1.2 equiv) in dry THF (5 mL). The reaction mixture was stirred at rt for 3 days. The mixture was filtered through glass wood, then a celite pad, and concentrated in vacuo. The crude product was washed with Et2O (10 mL 2) to remove the unreacted Btz affording 215 mg of crude product as a brown solid. Crude material was characterized by mass spectroscopy and exhibited no ion at 385.2 corresponding to Btz: Thermo-MS (ESI) (M + H) Calcd for C35H58BN6O12P2: 827.3. Found: 827.4. (M + Na) Calcd for C35H57BN6O12P2Na: 849.35. Found: 849.4. (M + K) Calcd for C35H57BN6O12P2K: 865.32. Found: 865.3. A dry 10 mL round bottom flask was charged with a solution of the above crude product (215 mg = 26 mmol) in dry CH2Cl2 (8 mL) under Ar at 0 C. Trimethylsilyl bromide (TMSBr) (239.7 mg, 0.156 mmol, 6 equiv) was added dropwise with magnetic stirring. After the addition was complete, the reaction mixture was allowed to stir and warm to rt overnight. The reaction mixture was then concentrated in vacuo and kept under high vacuum for 10 min to afford a brown solid. Then, methanol (5 mL) was added to dissolve the solid, and the resulting solution was concentrated in vacuo. This procedure was repeated 3 additional times. The resulting solid was dissolved in EtOH (0.5 mL), and this was followed by the addition of 10 mL of Et2O, resulting in the formation of a brownish-white precipitate. The solid was isolated by filtration and dried under high vacuum at 90 C affording 170 mg of 3 (BP-Btz) as a light brown solid (91%). Characterization Data for BP-Btz: 1H NMR (500 MHz, CDCl3): 8.95 (s, 1H), 8.68 (s, 1H), 8.62 (s, 1H), 7.37 (s, 4H), 7.16 (s, 1H), 4.71 (t, = 10 Hz, 1H), 4.30 (s, 2H), 3.94 (t, = 20 Hz, 1H), 3.74 (s, 4H), 3.47 (s, 2H), 3.28 (s, 4H), 3.08 (m, 1H), 2.97 (m, 1H), 2.78 (m, 1H), 1.32C1.24 (m, 2H), 1.16 (m, 1H), 0.72 (m, 6H). 13C NMR (400 MHz, CDCl3): 177.38, 163.85, 155.7, 148.34, 144.34, 144.14, 143.88, 135.23, 129.72, 128.77, 127.31, 59.59, 55.81, 55.68, 52.67, 50.67, 48.74, 47.88, 36.39, 24.83, 23.85, 21.99. 31P NMR (400 MHz, CDCl3): ?1.43, ?4.81. IR cm?1: 3275, 3055, 2954, 2146, 1716, 1701, 1676, 1629, 1521. TOF-MS (ES+) (M + 3H) Calcd for C27H44BN6O12P2: 717.26. Found: 717.55. Fendiline hydrochloride (M + 2H + Na) Calcd for C27H43BN6O12P2Na: 739.24. Found: 739.64. (M + H + 2Na) Calcd for C27H42BN6O12P2Na2: 761.22. Found: 761.59. HRMS (ESI) (M + Na) Calcd for C27H41BN6O12P2Na: 737.2241. Found: 737.2248. 2.1.3. 2-(bis(2-hydroxyethyl)amino)ethyl (bis(dihydroxyphosphoryl)methyl)carbamate (1 BP-Linker) An oven dried round bottom flask was charged with a solution of 0.1 g of Fendiline hydrochloride 2-(bis(2-hydroxyethyl)amino)ethyl(bis(diethoxyphosphoryl)methyl)carbamate (BP-Linker-) (0.209 mmol) in 1 mL anh CH2Cl2. After cooling to 0 C, neat bromotrimethyl silane (0.17 mL, 1.25 mmol) was added dropwise and the content was gradually warmed to room heat and stirred Rabbit polyclonal to AnnexinA11 for 16 h. The reaction mixture was concentrated under reduced pressure and treated with 5 mL of anh methanol. The alcoholic answer was concentrated and the procedure was repeated three times to yield a pale brown waxy solid. The crude mixture was dissolved in a methanol/water mixture and added to diethyl ether, whereupon 47 mg (62%) of 2-(((diphosphonomethyl)carbamoyl)oxy)= 2.4 Hz, 1H), 4.03C3.89 (m, 4H), 3.72C3.61 (m, 2H), 3.59C3.41 (m, 4H); 31P-NMR (400 MHz, CD3OD/D2O) 14.99;.