The oil-filled lipid NPs containing a DX-lipid conjugate with fine-tuned lipophilicity and activation kinetics effectively enhanced the therapeutic index of DX. The encouraging results of these studies suggest that the novel formulation holds guarantee for additional preclinical development.5. Experimental SectionMaterials and Animals: DX, PX, 2-bromohexadecanoic acid (99 ), 4-(dimethylamino) pyridine (DMAP) and N,N’-dicyclohexyl-carboiimide (DCC, 99 ) have been purchased 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 Bcl-W manufacturer polyethylene glycol-1000 succinate (Vitamin E TPGS) was bought from Eastman Chemicals (Kingsport, TN). BALB/c mouse plasma was bought from Revolutionary Investigation Inc. (Novi, MI). Sepharose CL-4B was purchased from GE Healthcare (Uppsala, Sweden). Hybrid-SPEcartridge 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 Form Culture Collection (ATCC) and have been maintained in RPMI-1640 medium with 10 fetal bovine serum (FBS). Female BALB/c mice, four to 5 weeks old, have been bought from Charles River (Wilmington, MA) and housed in a pathogen-free room. All experiments involving mice have been conducted 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 (0.62 g, 1.85 10-3 mol, 1.5N) and DCC (0.5 g, 2.47 10-3 mol, 2N) in dry CH2Cl2 (200 mL) beneath argon. The remedy 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) along with the reaction mixture was stirred at area temperature for an added five min. 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 through a fritted funnel, and the filtrate was evaporated beneath vaccuo. The crude product was purified by preparative TLC in CHCl3: MeOH (95:5). The silica gel was removed by filtration by means of a fine fritted funnel and the filtrate was evaporated below vaccuo to provide the preferred solution as a white CYP26 review powder (0.four mg, 86 ). 1H NMR (400 MHz, CDCl3): (ppm) = 0.8 (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, three), two.38 (s, 1H, 7), three.86 (d, 1H, four), 4.12 (d, 1H, two), 4.2 (t, 1H, HBrC1″), four.26 (t, 2H, 13), four.88 (d, 1H, ten), 5.2 (d, 2H, 20), 5.22 (d, 1H, 2′),Adv Healthc Mater. Author manuscript; accessible in PMC 2014 November 01.Feng et al.Page5.62 (d, 1H, 3′), 7.22.53 (m, 8H, r-H268 and Ar-H305), 8.05 (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 ( six,14), 43.1 ( 15), 44.five ( 3), 45 ( HBr), 46.4 ( 3′), 57.5 ( 8), 71.eight ( 13), 72.1 ( 7), 74.four ( two), 75 ( ten), 75.3 ( 20), 78.9 ( 6′), 79.9 ( 1), 80.9 (C4), 84.2 ( five), 126.3 ( 31,33,35), 128.9 ( 32,34), 129.2 ( 26,28), 130.two ( 24,25,29), 133.six ( 27), 135.5 ( 11), 138.9 ( 12), 154.2 ( 5′), 167 ( 23), 16.