Antitumor Activity And Mechanism Of The Novel Platinum Compound TCP-02

Objective:Platinum anticancer drug is one of the most representative medicines in tumor therapeutic area,search for new platinum based chemotherapy has been one of the hot research in the field of anti tumor.This dissertation by using the structure of cyclohexanediamine-platinum to couple GA and its derivatives,design and synthesis of a series of TCP compounds,through the research of its antitumor activity in vivo and in vitro,pharmacokinetic parameters and mechanism of action,in order to obtain a new platinum anticancer drugs involving the higher antitumor activity.Methods:1.Using MTS colorimetric method to screen the antitumor activity of the TCP series compounds in vitro to select the target compound（TCP-02）,and using the same method to detect the proliferation inhibition rates of TCP-02 on eight different tumor cells,so as to evaluate its antitumor activity in vitro.2.Adopting the animal model of tumor-bearing mice to evaluate the antitumor activity of TCP-02 in vivo.The tumor vaccine after 24 h and randomly divided them into four groups,namely model group,positive drug cisplatin group,high and low TCP-02 concentration group,continuous dosing for 10 d,through measuring the rate of life extension and the change index of abdominal perimeter to evaluate the antitumor activity of TCP-02 in vivo.3.Flow cytometry as the main detection means to study the antitumor mechanism of TCP-02.Using PI reagent to detect the influence of TCP-02 on Hep G2cells cycle,employing the phosphatidylserine eversion method to detect the influence on apoptosis of Hep G2,and detecting the across mitochondrial membrane potential and the intracellular ROS level of cells,to evaluate the role on mitochondrial dysfunction in the process of TCP-02 induced cell apoptosis of Hep G2.4.Studying the pharmacokinetic parameters change of the SD rats after intravenous given TCP-02 in vivo,analyzing the blood drug concentration dynamic change rules and characteristics of liver tissue distribution of TCP-02 in animals,preliminary observing the pharmacokinetic characteristics of TCP-02 in vivo.Results:1.The TCP-02 was selected as the most obvious target compound,and further study showed that the TCP-02 has better inhibition on proliferation of liver cancer,lung cancer,breast cancer,cervical cancer and other tumor cells,and all the IC₅₀values were around 10μmol·L^-1,among those,the inhibiting effect on liver cancer cell proliferation was relatively strong,and the IC₅₀ value was 8.67μmol·L^-1.2.According to the results of Dutch S180 ascites tumor mouse model to evaluate the antitumor activity of TCP-02 in vivo,the change of abdominal perimeter and life extension rate in mice were statistically significant（P<0.01）,which indicated that TCP-02 with a significant inhibitory effect on the formation of ascites tumor in mice.Besides,the life extension rate of the TCP-02 group was significantly higher than that of the cisplatin group with the same dose.3.When TCP-02 function on Hep G2 cells at the concentrations of IC₅₀,could appear a significant G2/M retardant on cell cycle after 36 h,and has obvious dose dependent.TCP-02 function for 24 h,the apoptosis of Hep G2 cells could be induced by dose-dependent.And the reduction of the membrane potential of mitochondria and the sustained release of ROS maybe the important reasons for inducing apoptosis in the process of TCP-02.4.The pharmacokinetic parameters in vivo of TCP-02 in rats showed it widely distributed in the body,eliminated slowly,and the elimination half-life was 11 h.The concentration of prototype drug in liver tissue was 20 to 50 times than it in the blood,and can maintain a higher drug concentration in the liver tissue for a long time.Conclusions:TCP-02 has antitumor activity both in vivo and in vitro,and has different mechanism compares with cisplatin,and extensive distribution and elimination in rat,and can be kept higher drug concentration for a long time in liver tissue.Consequently,TCP-02 possesses excellent potential medicinal prospects for liver cancer and is expected to become a new kind of valuable platinum drug.