Inhibitory Effect Of Irinotecan Hydroclorjde-Dihydromyricetin Compound Liposomes On Liver Cancer Cells And Its Pharmacokinetics

[Object]:To explore the combination of Irinotecan(CPT-11)and Dihydromyricetin(DMY),To study the effect of compound liposome of CPT-11 and DMY on HepG-2 cells;Through the preparation of the compound liposome of CPT-11 and DMY,its morphology,particle size distribution,entrapment rate,drug loading were explored;To investigate its release behavior in vitro and its freeze-drying process;To study the effect of the compound liposome of CPT-11 and DMY on HepG-2 cells;The pharmacokinetic behavior after intravenous injection and administrations-of rats tail was studied,and the related parameters were calculated.[Method]:Screend the effective concentration range of CPT-11 and DMY against tumor cells in vitro by MTT,and to determine the optimal proportion of HepG-2.The cell inhibition of liposome against HepG-2 was investigated by MTT method;CPT-11-DMY-Lip solution was prepared by the ethanol injection method and ammonium sulfate gradient method.According to L9(34)orthogonal test,the drug prescription was optimized,and the encapsulation efficiency,drug loading,particle size distribution and appearance morphology were taken as indexes to futher optimize the drug prescription,determine the best technology and verify it;Particle size analyzer determine the potential of the compound liposomes,particle size and distribution;The freeze-drying process was optimized with appearance,color,redispersibility and Particle size as the indicators;The morphology of liposome was observed by transmission electron microscopy;Through hemolytic test to examine the preparation can lead to rupture of red blood cells,causing hemolysis;The in vitro releasing tendency of CPT-11-DMY-Lip was analyzed by dynamic dialysis method;To study the effect of the compound liposome of CPT-11 and DMY on HepG-2 cells;The pharmacokinetic parameters were calculated by DAS2.0 software and analyzed using SPSS 17.0 statistic software.[Results]:The quality ratio of CPT-11 to DMY was determined to be 1:4,The combination of the two has a synergistic effect;In the CPT-11-DMY-Lip prepared by screening the optimum formulation with orthogonal experiment,the entrapment rate of CPT-11 reached(8 2.5 8± 1.4 8)%,and its drug loading reached(4.5±1.3)%;the average encapsulation efficiency of DMY reached(71.45± 1.03)%,and its drug loading reached(5.7±1.1)%;The determination results of particle size analyzer showed that the particle size haduniform distribution,with average particle size of(123.1±1.8)nm;the PDI value was 0.181;and the Zeta electrical potential was(-24.3±0.51)mV;7%mannitol was chosen as lyoprotectant.The appearance of lyophilized samples was loose.After adding water for injection,the dispersibility was good.The liposome solution was milky white with blue opalescence;The stability of freeze-dried powder experiments found that the compound liposomes at 4℃ for 6 months.The particle size decreased by 5.8nm,the entrapment efficiency decreased by 6.9%and 8.3%,and the stability was good;In the water for injection,the freeze-drying CPT-11-DMY-Lip had good redispersibility,and from the observation of transmission electron microscope,CPT-11-DMY-Lip had the spherical shape and uniform size;There were no phenomena of hemolysis and flocculation occurring in the injection-use freeze-drying powder;The results of in vitro fitting showed that the release behavior of the composite liposomes was consistent with the Weibull model.The release of 2%Tween-80 in PBS buffer(pH6.5)resulted in the cumulative release rates of CPT-11 and DMY being 61.45±1.88%and 72.67±1.07%;The results of in vitro cytology showed that the compound liposomes could inhibit the normal growth of tumor cells.After a series of concentrations of drug were applied to HepG2 cells for 48h,the inhibitory rate of compound liposomes was higher than that of the same drug concentration;According to the pharmacokinetic parameters of rats,the half-lives of the liposomes were longer than those of the free drugs at the same dose,which had a strong sustained-release effect.[Conclusion]The preparation of CPT-11-DMY-Lip compound liposome by the ethanol injection method method and ammonium sulfate gradient method is simple and convenient;The average particle sizes were small and uniformLy distributed,and had good morphology;the electrical potential was stable,and the encapsulation rate was high;CPT-11 is encapsulated in a hydrophilic inner layer and DMY is encapsulated in a hydrophobic outer layer;In vitro anti-tumor study,CPT-11-DMY compound liposomes on HepG2 cells have a significant effect of promoting apoptosis;CPT-11-DMY compound liposomes in vivo pharmacokinetic studies in higher bioavailability,reducing plasma clearance.