Conjugate with fine-tuned lipophilicity and activation kinetics successfully enhanced the therapeutic index of DX. The encouraging benefits of these research suggest that the novel formulation holds promise for additional preclinical improvement.five. Experimental SectionMaterials and Animals: DX, PX, 2-bromohexadecanoic acid (99 ), 4-(dimethylamino) pyridine (DMAP) and N,N’-dicyclohexyl-carboiimide (DCC, 99 ) have been bought from Sigma-Aldrich (St. Louis, MO). Miglyol 808 was obtained from Sasol (Witten, Germany). Polyoxyl 20-stearyl ether (Brij 78) was obtained from Uniqema (Wilmington, DE). D-alphatocopheryl polyethylene glycol-1000 succinate (Vitamin E TPGS) was bought from Eastman Chemicals (Kingsport, TN). BALB/c mouse plasma was bought from Innovative Research Inc. (Novi, MI). Sepharose CL-4B was purchased from GE Healthcare (Uppsala, Sweden). Hybrid-SPE?cartridge was bought from Sigma-Aldrich Supelco (St. Louis, MO). The human prostate cancer cell line DU-145, and murine breast cancer cell line 4T1 had been obtained from American Type Culture Collection (ATCC) and have been maintained in RPMI-1640 medium with 10 fetal bovine serum (FBS). Female BALB/c mice, four to five weeks old, had been purchased from Charles River (Wilmington, MA) and housed in a pathogen-free area. All experiments involving mice have been performed according to an authorized animal protocol by the University of North Carolina Institutional Animal Care and Use Committee. General procedure for the synthesis of 2′-(2-bromohexadecanoyl)-docetaxel (2-Br-C16DX)[7] A flame-dried round-bottom flask was charged with (?-2-bromohexadecanoic acid (0.1373253-24-7 Data Sheet 62 g, 1.(R)-2-Methylazetidine hydrochloride Purity 85 ?10-3 mol, 1.5N) and DCC (0.5 g, two.47 ?10-3 mol, 2N) in dry CH2Cl2 (200 mL) beneath argon. The resolution was stirred for 10 min at space temperature. DX (1.0 g, 1.24 ?10-3 mol, 1N) was added as well as a catalytic amount of DMAP (0.15 g, 1.24 ?10-3 mol, 1N) and also the reaction mixture was stirred at area temperature for an extra five min.PMID:25804060 The reaction was monitored by TLC (CH2Cl2: MeOH 95:5 v/v; 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 product was purified by preparative TLC in CHCl3: MeOH (95:5). The silica gel was removed by filtration via a fine fritted funnel along with the filtrate was evaporated beneath vaccuo to provide the desired product as a white powder (0.4 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), 2.24 (m, 1H, three), two.38 (s, 1H, 7), three.86 (d, 1H, four), 4.12 (d, 1H, 2), four.2 (t, 1H, HBrC1″), 4.26 (t, 2H, 13), 4.88 (d, 1H, 10), five.2 (d, 2H, 20), five.22 (d, 1H, 2′),Adv Healthc Mater. Author manuscript; offered in PMC 2014 November 01.Feng et al.Page5.62 (d, 1H, 3′), 7.22?.53 (m, 8H, r-H26?eight and Ar-H30?5), 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.six (?(CH2)14C1″), 31.9 ( 6,14), 43.1 ( 15), 44.five ( 3), 45 ( HBr), 46.4 ( 3′), 57.5 ( eight), 71.8 ( 13), 72.1 ( 7), 74.four ( two), 75 ( 10), 75.3 ( 20), 78.9 ( 6′), 79.9 ( 1), 80.9 (?C4), 84.two ( five), 126.three ( 31,33,35), 128.9 ( 32,34), 129.two ( 26,28), 130.2 ( 24,25,29), 133.six ( 27), 135.5 ( 11), 138.9 ( 12), 154.2 ( 5′), 167 ( 23), 167.three ( 21), 169 ( 1), 169.7 ( 1.