Preparation And Characterization Of Glycolysis Inhibitor 3BP Coupled Temperature-sensitive Nanogel

Primary Liver Cancer(PLC)has become the sixth highest-incidence cancer in the world,and its mortality rate is second only to lung cancer and colorectal cancer,and China is a region with a high incidence of liver cancer.For patients with advanced liver cancer,transcatheter arterial chemoembolization(TACE)is often used clinically for treatment.The therapeutic effect of TACE is closely related to the fluidity and embolization of the vascular embolization agent.The temperature-sensitive nanogel embolization agent developed in the early stage of our research group takes into account both fluidity and embolism,which is an ideal vascular embolization agent.Aiming at the problem of aggravating the hypoxic microenvironment at the tumor site after TACE,glycolysis inhibitor is combined with temperature-sensitive nanogels in this article,and they are applied in TACE treatment to achieve the purpose of improving the hypoxic microenvironment.3-Bromopyruvate(3BP),as a typical glycolysis inhibitor,shows obvious anti-tumor effects and is tumor-specific.In this paper,based on the temperature-sensitive nanogel(PNH),3BP is connected to the nanogel by chemical coupling to prepare nanogels with glycolysis inhibitor function,namely 3BP-CO-PNH(ester bond connection)and 3BP-CN-PNH(acylhydrazone bond connection).Under the stimulation of cell esterase and acidic environment,3BP can be released from these two drug-loaded nanogels and exert glycolysis inhibitory effect.Through the NMR characterization of the nanogels before and after drug loading,it was determined that 3BP was coupled to the PNH nanogels by means of ester bond and acylhydrazone bond,respectively.The introduction of the hydrophilic molecule 3BP has no effect on the morphology and particle size of the nanogels,but it has an effect on the zeta potential of the PNH nanogels at 37 ℃,and makes the volume phase transition temperature(VPTT)and the lower critical solution temperature(LCST)of 3BP-CO-PNH and 3BP-CN-PNH have a certain increase.Because the temperature range of VPTT and LCST is 32 ～ 34 ℃,3BPCO-PNH and 3BP-CN-PNH can be used as a vascular embolization agent in TACE treatment.In addition,cytotoxicity experiments show that 3BP-CO-PNH and 3BP-CN-PNH nanogels have strong cytotoxicity to HepG2 cells,have certain biological safety to HUVEC cells,and can significantly reduce the ATP expression level of tumor cells.In summary,in this paper,two temperature-sensitive nanogels coupled with glycolysis inhibitor 3BP were prepared,and the structure,morphology,performance and effect evaluation in vitro of the two were studied.The VPTT and LCST of 3BP-CO-PNH and3BP-CN-PNH are close to human body temperature and are suitable as vascular embolization agents for liver cancer.Both of them also show obvious killing effects on tumor cells,laying the foundation for their application in TACE therapy.