Fabrication of Multifunctional Targeted Icg-Doxorubicin- Incorporated Perfluorinated Nano-Theranostic Agents for Fluorescence Diffuse Optical Imaging and Photo-/Chemo-Therapy of Breast Cancer (Ii)(Iii)

Specific Aim: We aim to design and fabricate multifunctional targeted Indocyanine green(ICG)-Doxorubicin (DOX) incorporated perfluorinated nanoemulsions (TIDPNEs), and explore theirpotential for uses in 1) photodynamic-, 2) photothermal-, 3) chemo-therapies, and 4) near-infrared (NIR)fluorescence diffuse optical tomography (FDOT).Background: Theranostic agent has been demonstrated as an emerging and promising therapeutic strategyfor cancer resistance. DOX is one of the most commonly used anticancer drugs. However, serious side effectscaused from high dose hamper its application. In terms of the cancer diagnostics, FDOT has been widelyinvestigated especially in clinical detection of breast cancer due to its noninvasive accessibility, highsensitivity, and easy-to-use capability, whereas ICG, a water-soluble tricarbocyanine dye, is one of fewclinically proved NIR fluorophores. Recently ICG is further used as the agent of photo-therapy for cancertreatment since ICG can produce singlet oxygen and heat upon NIR light irradiation. However, manydrawbacks of ICG such as 1) concentration-dependent aggregation, 2) irreversible and accelerateddegradation in aqueous solution, and 3) short half-life (3 – 4 min) in vivo tremendously hamper itsapplicability. Therefore, development of a targeted ICG-DOX incorporated probe with high stability andeffectiveness is urgently needed for breast cancer resistance.Features: To enhance the applicability of the developed theranostic agent, TIDPNEs were assembled byusing perfluorocarbon liquid, ICG, and DOX molecules that enable to provide following advantages:[1] Multifunctional TIDPNEs can provide both diagnosis and treatment for breast cancer cells where thedestruction of cancer cells can be achieved by physical and chemical approaches.[2] ICG and DOX molecules can be protected by the emulsions. Furthermore, the developed TIDPNEs withhigh binding specificity may enhance the amount and efficiency of ICG and DOX delivery to the targetcancer cells. Therefore, the drug dose can be diminished and the stability of ICG can be improved.[3] Since perfluorocarbon possesses high O2-dissolving capability, the developed TIDPNEs enable to provideabundance of oxygen molecules for photodynamic therapy.[4] Based on the binding specificity of the TIDPNEs, ICG and DOX molecules can be concentrated on thetarget cells and therefore the efficacy of imaging diagnosis, photo-, and chemo-therapy can be raised.Research Schedule: The following tasks will be performed orderly in the next two years: Stage 1: Optimization and characterization of TIDPNEs (The 1st Year)[1] To evaluate the loading efficiency, content percentage, degradation rate and drug release efficiency forencapsulated ICG and/or DOX under static setting without laser irradiation[2] Cytotoxicity of the TIDPNEs[3] Capability of oxygen carry of TIDPNEs[4] Binding specificity of the TIDPNEs for breast cancer cells Stage 2: Applicability of the TIDPNEs for imaging, photo- and chemotherapy (The 1st Year)[1] Efficacy of TIDPNEs on NIR fluorescent expression[2] Effect of TIDPNEs on FDOT analysis[3] Effect of TIDPNEs on singlet oxygen generation[4] Effect of TIDPNEs on temperature increase[5] Release efficiency of encapsulated DOX molecules[6] Optimization of TIDPNEs for FDOT and therapeutic applications Stage 3: In vitro and in vivo examinations for the developed TIDPNEs (The 1st - 2nd Year)[1] Cell assay using human breast cancer cell line (in vitro assay)[2] Animal test. Modification of the TIDPNEs and reproducibility examination (in vivo assay)