Development And Properties Of Multifunctional IR780-based Derivates

Objective:To develop novel multifunctional IR780-based derivatives and explore their potential application in biomedicine area.Method:Two novel IR780-based derivatives?L₁and L₂?with different chain length are prepared by the substitution reaction of indocyanine core IR780 and alkylmorpholine?2-aminoethylmorpholine,3-aminopropylmorpholine?.Modern analytical methods,including ¹H-NMR,¹³C-NMR,high resolution mass spectrometry,FT-IR,are used to characterize the structures of L₁ and L₂.Ultraviolet-visible absorption and fluorescence spectrophotometer are used to study their in vitro spectroscopic properties and selective hypochlorous acid recognition abilities.DPBF method is employed to detect in vitro singlet oxygen production under 660 nm light.Quantum chemical calculations of two IR780-based derivatives and the control compound IR780 are performed by Gaussian09software.NMR titration,mass spectrometry test,and spectroscopic analysis are performed to evaluate the photostability of the IR780-based derivatives.At the cellular level,MTT assay is used to examine the darktoxicity and phototoxicity of L₁ and L₂ on HepG2?B16F10 two cancer cell lines as well as the darktoxicity of RAW 264.7 cells.Fluorescence inverted microscop is used to verify properties of mitochondria-?lysome-targeting ability in HepG2?B16F10 and RAW 264.7 cells as well as the properties of endogenous and exogenous hypochlorous acid recognition in A549 and RAW 264.7 cells were studied.At the living body level,tail vein injection is first used to evaluate the tumor-targeting property,photodynamic antitumor activity,and NIR fluorescence imaging performance of L₁ and L₂in C57 mice bearing with B16F10 tumor model.The in vivo safety of L₁ and L₂ are examined by H&E staining of the main organs such as heart,liver,spleen,lung,and kidney,together with the determination of liver and kidney function-related serum indexes.Further,the properties of hypochlorous acid near-infrared fluorescence imaging are evaluated by near-infrared fluorescence imaging in a mouse model of LPS/D-GalN-induced acute liver injury and LPS-induced leg inflammation.Results:The exact molecular structures of L₁and L₂ were confirmed by ¹H-NMR,¹³C-NMR,FT-IR and high resolution mass spectrometry.Studies at the molecular,cellular and living levels have shown that:?1?the two target molecules have improved photostability,dual tumor targeting properties,and near-infrared fluorescence imaging and PDT-targeting properties compared to IR780.?2?two target molecules can selectively recognize hypochlorous acid with"On-Off"fluorescence response and perform near-infrared fluorescence imaging fo hypochlorous acid in mouse models of LPS/D-GalN-induced acute liver injury and LPS-induced leg inflammation.?3?preliminary in vivo safety studies also shows that they have no obvious toxicity to common organelles and tissues.Conclusion:Two novel multifunctional IR780-based derivatives with near-infrared fluorescence imaging diagnosis-mediated photodynamic therapy and highly selective recognition of hypochlorous acid have been obtained.The studies in this thesis provides new design ideas and research references.