Ocetaxel (2-Br-C16DX)[7] A flame-dried round-bottom flask was charged with (-
Ocetaxel (2-Br-C16DX)[7] A flame-dried round-bottom flask was charged with (-2-bromohexadecanoic acid (0.62 g, 1.85 10-3 mol, 1.5N) and DCC (0.5 g, two.47 10-3 mol, 2N) in dry CH2Cl2 (200 mL) GSK-3α drug beneath argon. The resolution was stirred for ten min at area temperature. DX (1.0 g, 1.24 10-3 mol, 1N) was added as well as a catalytic volume of DMAP (0.15 g, 1.24 10-3 mol, 1N) and the reaction mixture was stirred at area temperature for an further five min. The reaction was monitored by TLC (CH2Cl2: MeOH 95:5 vv; Rf = 0.58) for completion. The white precipitate of dicyclohexyl urea byproduct was filtered by way of a fritted funnel, and also the filtrate was evaporated below vaccuo. The crude solution was purified by preparative TLC in CHCl3: MeOH (95:5). The silica gel was removed by filtration by means of a fine fritted funnel as well as the filtrate was evaporated below vaccuo to give the preferred solution as a white powder (0.four mg, 86 ). 1H NMR (400 MHz, CDCl3): (ppm) = 0.eight (t, 3H, H3(CH2)14), 1.05 (s, 6H, 16,17), 1.16 (s, 9H, 7”), 1.19 (s, 3H, 19), 1.23 (m, 28H, (CH2)14CH3), 1.68 (s, 3H, 18), 1.78 (m, 2H, 14), 1.67 (d, 2H, H2C1″), 1.87 (s, 3H, H22), two.24 (m, 1H, 3), two.38 (s, 1H, 7), 3.86 (d, 1H, 4), 4.12 (d, 1H, two), four.two (t, 1H, HBrC1″), 4.26 (t, 2H, 13), 4.88 (d, 1H, 10), five.two (d, 2H, 20), 5.22 (d, 1H, 2′),Adv Healthc Mater. Author manuscript; obtainable in PMC 2014 November 01.Feng et al.Page5.62 (d, 1H, 3′), 7.22.53 (m, 8H, r-H268 and Ar-H305), eight.05 (d, 2H, rH25,29). 13C NMR (100 MHz, CD3OD): (ppm) = eight.9 ( 19), 14.1 ( H3(CH2)20), 20.9 (C18), 22.6 ( 22), 23.7 (CH2)19CH2CH3), 27 ( 16,17), 28.1 ( 7”), 29.6 ((CH2)14C1″), 31.9 ( six,14), 43.1 ( 15), 44.5 ( 3), 45 ( HBr), 46.four ( 3′), 57.5 ( eight), 71.eight ( 13), 72.1 ( 7), 74.four ( 2), 75 ( ten), 75.three ( 20), 78.9 ( 6′), 79.9 ( 1), 80.9 (C4), 84.two ( 5), 126.3 ( 31,33,35), 128.9 ( 32,34), 129.two ( 26,28), 130.2 ( 24,25,29), 133.six ( 27), 135.five ( 11), 138.9 ( 12), 154.2 ( 5′), 167 ( 23), 167.three ( 21), 169 ( 1), 169.7 ( 1″), 211.five ( 9). Characterization of DX and DX conjugates Electrospray Ionization (ESI) coupled with direct injection was employed to figure out the mz of your final synthetic conjugate solution by Thermo Scientific TSQ Quantum Access with positive ionization. The mz in the observed molecular ion was 1125, which clearly corresponded for the H adduct of 2-Br-C16-DX. The 2-Br-C16-DX concentrations were quantified by HPLC working with a Finnigan Surveyor HPLC program with a Photodiode Array (PDA) detector, autosampler and LC pump plus having a InertsilODS-3 column (4 , 4.six 150 mm, GL Sciences) at 25 . Chromatographic separation was achieved by gradient elution working with mobile phase 2-propanol, acetonitrile (ACN) and water (five: 55: 40 vvv). The flow rate was 1.0 mLmin plus the total run time was 25 min for each 25 injection. The wavelength was 230 nm. The DX concentration was quantified by LCMSMS as described previously.[4] 2-Br-C16-DX digestion in fresh mouse plasma The esterase digestion study was performed in fresh BALBc mouse plasma. The 2-Br-C16DX NPs (0.five mgmL) were spiked into the plasma to create a final concentration of 10 mL. The mixture was incubated at 37 inside a water bath shaker. At designated time points, 100 of digestion mixture was removed. The concentration of 2-Br-C16-DX was determined by DP Purity & Documentation Hybrid-SPE precipitate technique as described previously followed by HPLC evaluation.[4] The 2-Br-C16-DX remaining at any time point was calculated as one hundred the ratio of remaining drug amount towards the total drug spiked into this.