Application Of Near-infrared Fluorescent Dye IR-780 Coupled With Cabazitaxel In The Diagnosis And Treatment Of Renal Cell Carcinoma And Prostate Cancer

Background:Renal cell carcinoma（RCC）is the most common malignant tumor of urinary system.The disease is insidious and develops rapidly.About 15%of the patients have metastasis at the time of diagnosis and miss the best opportunity of surgical treatment.The prognosis of metastatic renal cell carcinoma（m RCC）is poor.Systemic drug treatment can benefit the survival of patients,but some of the m RCC patients are not sensitive to the treatment,and some of the patients get drug resistant after treatment.Therefore,early detection,diagnosis and resolution of drug resistance become the key to the treatment of RCC.Prostate cancer（PCa）is the most common malignant tumor of male genitourinary system.In recent years,its morbidity and mortality are increasing in China.The main cause of death of PCa patients is the progression of high-risk PCa to castration resistant prostate cancer（CRPC）after treatment.CRPC patients have high drug resistance and poor prognosis.Early diagnosis and resolution of drug resistance is also the key to PCa treatment.Near infrared fluorescence（NIRF）dye IR-780 has tumor targeting,which can be efficiently absorbed by tumor cells and gathered in tumor tissue,and make tumor tissue imaging under the condition of near infrared（NIR）.Therefore,we take advantage of this feature of IR-780 and combine it with the antitumor drug cabazitaxel to synthesize a new drug Caba-780 which should be used in the diagnosis and treatment of RCC and CRPC.Method:1.A new drug named Caba-780 was formed by combining IR-780 with cabazitaxel by chemical coupling.The characterization structure of Caba-780 was verified by infrared spectroscopy,nuclear magnetic resonance spectroscopy（NMR）,and high-resolution mass spectra（HRMS）.2.Cell experiments were carried out on two RCC cells（ACHN and 786-O）and normal human embryonic kidney cell line HEK293.The uptake and subcellular localization of Caba-780 by RCC cells and normal cells were observed by laser confocal microscopy.The antitumor activity of Caba-780 in vitro was evaluated by cytotoxic assay,plate cloning assay,scratch migration assay,Transwell chamber invasion assay and cell flow cytometry.ACHN cells were used to establish xenograft tumor bearing nude mice model,and the tumor imaging and antitumor effect of Caba-780 in vivo were evaluated by animal experiments.3.Another cell experiments were carried out on two CRPC cells（DU145 and PC-3）and human normal prostate stromal immortalized cell WPMY-1.The uptake and subcellular localization of Caba-780 by CRPC cells and normal cells were observed by laser confocal microscopy.The antitumor activity of Caba-780 in vitro was evaluated by cytotoxic assay,plate cloning assay,scratch migration assay,Transwell chamber invasion assay and cell flow cytometry.DU145 and PC-3 cells were used to establish xenograft tumor bearing nude mice model,and the tumor imaging and antitumor effect of Caba-780in vivo were evaluated by animal experiments.Results:1.The synthetic new drug Caba-780 is a lake-blue solid powder under normal temperature and pressure,which is fat-soluble.Its molecular formula is C₈₇H₁₀₈N₅O₁₆and molecular weight is 1507.7689 g/mol.Its excitation,ultraviolet absorption and fluorescence spectra are all located in the NIR region.Caba-780 has good biocompatibility and stable imaging in the NIR region.2.Caba-780 can be efficiently absorbed by ACHN and 786-O cells and the absorbed Caba-780 is located on the lysosome and mitochondria of RCC cells.In vitro,Caba-780can significantly inhibit the proliferation,cloning,migration and invasion of ACHN and786-O cells,and can promote the apoptosis of ACHN and 786-O cells by inhibiting G1phase and prolonging G2 phase in cell cycle.Moreover,Caba-780 has stronger anti RCC cell ability than IR-780 and cabazitaxel.In vivo,Caba-780 can gather in RCC tissue,make RCC tissue imaging under NIR condition,and also can significantly inhibit the growth of RCC tissue.3.Caba-780 can be efficiently absorbed by DU145 and PC-3 cells in a concentration and time-dependent manner.The ingested Caba-780 is located in lysosomes and mitochondria of CRPC cells.In vitro,Caba-780 can significantly inhibit the proliferation,cloning,migration and invasion of DU145 and PC-3 cells,and can promote the apoptosis of DU145 and PC-3 cells by inhibiting G1 phase and prolonging G2 phase in cell cycle.In addition,Caba-780 has stronger anti-CRPC cells ability in vitro than IR-780 and cabazitaxel.In vivo,caba-780 can gather in CRPC tissue,make CRPC tissue imaging under NIR condition,and also can significantly inhibit the growth of CRPC tissue.Conclusion:The new synthetic drug Caba-780 has the characteristics of NIRF.It can be efficiently absorbed by RCC and CRPC cells in vitro and can inhibit the proliferation and promote the apoptosis of RCC and CRPC cells by inhibiting G1 phase and prolonging G2 phase in cell cycle.In vivo,Caba-780 can gather in RCC and CRPC tissues,imaging tumor tissues under NIR condition,and can significantly inhibit the growth of both tumor tissues.The new drug Caba-780 has a certain application prospect both in the diagnosis and treatment of RCC and CRPC.