| The proposed research focuses on the following two areas, one is to synthesize gold nanorods (GNR) and conjugates the antibody with nanorods, and characterize the properties and stabilities of the conjugates; to evaluate the binding specific, affinity and thermal ablation efficacy of the resulting conjugates in vitro assays; in vivo photothermal treatments performed on tumor bearing mice by intravenous injection of the GNR conjugates. The other one is to develop itraconazole (ITZ)-loaded Poly (lactic- co-glycolic acid) (PLGA) nanospheres (PLGA-ITZ-NS) around 200 nm with high encapsulation efficiency as a novel formulation for intravenous ITZ administration. On the first study, synthesis of gold nanorods according to the seed-mediated growth method, conjugation of GNR with PEG and anti-CA IX (PEG-GNR-anti-CA IX), characterization of the conjugates, evaluation of the binding affinity and then sandwich ELISA experiments were performed by coating the 96-well plates with CA IX antigen. Thermal ablation of HT29 cell lines and tumor bearing mice with HT29 cells were conducted. On the second study, preparation of PLGA-ITZ-NS by the nanoprecipitation and evaporation methods, characterization and lyophilization of PLGA-ITZ-NS; Developed and validated of analytical methods (including high performance liquid chromatography [HPLC] and liquid chromatography-tandem mass spectrometry [LCMS/MS]); pharmacokinetic study was performed by comparing the bio exposure between PLGA-ITZ-NS with SporanoxRTM. We demonstrated both the selectively photothermaI destruction of CA IX-positive cancer cells in vitro and CA IX-positive tumors in vivo using GNR/anti-CA IX conjugates. The optimal PLGA-ITZ-NS formulation has a similar bio exposure to Sporanox while providing a sustained plasma level (100 ng/mL) for up to 24 hours after intravenous administration. In the future, GNR conjugating with anti-CA IX could be a therapeutic nanoconstruct for efficient tumor targeting and photothermal therapy. Our newly developed PLGA-ITZ-NS has shown great sustained release and comparable bio exposure with Sporanox, therefore having the potential to be an alternative injectable formulation of ITZ. |
