Study On Pharmacokinetics,Tissue Distribution And Acute Toxicity Of Cisplatin-loaded Sterically Stabilized And Targeted Liposome

Objective:Cisplatin is a cell cycle non-specific drug that is used clinically as a first-line chemotherapy drug for the treatment of multiple solid tumors.However,cisplatin administration causes various dose-limiting toxic reactions,the most important of which is nephrotoxicity.To improve the targeting and reduce toxic and side effects of cisplatin treatment,we developed a cisplatin-loaded sterically stabilized and targeted liposome?ES-SSL-DDP?that has been shown effectively target to Hela cells and cervical tumors which positively express estrogen receptors,and has a significant anti-tumor effect.At present,in order to elucidate the distribution,metabolism and toxicity of ES-SSL-DDP in vivo,the pharmacokinetics,tissue distribution and acute toxicity of this cisplatin long-circulating targeted liposome were studied.Methods:?1?Establish and validate in vivo analysis methods of cisplatin.Pre-column derivatization HPLC was used to complex the cisplatin with sodium diethyldithiocarbamate?DDTC?.The resulting Pt?DDTC?₂ exhibited strong UV absorption at 254 nm.The HPLC method uses methanol/water?75:25?as the mobile phase and a flow rate of 1 mL/min.?2?Injection of 6.0 mg/kg dose of free cisplatin?DDP?,normal cisplatin liposome?L-DDP?and cisplatin-loaded sterically stabilized and targeted liposome?ES-SSL-DDP?through mouse tail vein.The blood and tissue samples were taken at different time points.After treatment,the plasma concentration and tissue content of cisplatin were determined by HPLC.The pharmacokinetic parameters were calculated using DAS 2.0 software to investigate the pharmacokinetics and tissue distribution.?3?After mice were injected with different doses of DDP,L-DDP and ES-SSL-DDP,the state of the animals and the number of deaths were observed and recorded for 14 days.The LD₅₀ was calculated and whole blood analysis was performed on the 14th day surviving mice.The viscera coefficient was calculated and the blood urea nitrogen and creatinine content were measured.Then the acute toxicity of ES-SSL-DDP was evaluated comprehensively.Results:?1?The retention time of cisplatin was 8.22 min.There was no impurity interference and the resolution was good?R>1.5?.The calibration curves for plasma and tissue samples showed good linearity with linear ranges of 0.1 to 10.0?g/mL.The minimum limit of detection and detection limit were 0.05?g/mL and 0.1?g/mL,respectively.The intra-day and inter-day precision of plasma samples were all less than 10%,and the recoveries were between 100 and 110%.The precision of the tissue samples were less than 10%,and the recoveries were between 90 and 110%.The stability of the sample after treatment indicates that the sample must be processed within 12 h after treatment.?2?Compared to free DDP,ES-SSL-DDP showed a2.4-fold and 2-fold increase in C_max and t_1/2,respectively,and a 25-fold increase in AUC.The tissue distribution of ES-SSL-DDP is different from that of free DDP.ES-SSL-DDP has a higher distribution in heart and spleen tissues and is less distributed in kidney tissue than DDP group.?3?The LD₅₀ of DDP,L-DDP and ES-SSL-DDP injected into the tail vein of mice was 13.1,16.1 and 18.0 mg/kg,respectively.Compared to free DDP and L-DDP,mice injected tail vein with ES-SSL-DDP had better survival and weight recovery at the same dose.There was no significant change in whole blood analysis and organ coefficient in the ES-SSL-DDP group.BUN and Cre levels in the free DDP group increased significantly,while the BUN and Cre levels in the ES-SSL-DDP group and L-DDP group were comparable to those in the normal saline group.Conclusion:Conclusion:In this paper,a method for in vivo analysis of cisplatin was successfully established by high performance liquid chromatography.The HPLC method meets the methodological requirements and can be used for the determination of cisplatin in plasma and tissue samples.The mouse tail vein injection of cisplatin-loaded sterically stabilized and targeted liposome?ES-SSL-DDP?has higher plasma concentration and longer half-life,indicating that ES-SSL-DDP can slow the metabolic elimination rate of cisplatin,prolong the circulation time of cisplatin in the body.ES-SSL-DDP can alter the distribution of cisplatin tissue and reduce the accumulation of cisplatin in the kidney.ES-SSL-DDP has less acute toxicity and reduces the toxicity to the blood system and nephrotoxicity of cisplatin.