Preparation Of Formulation Of Liver Cancer-Targeted DOX-LPD-polyplexes And Evaluation In Vivo

Objective:Malignant tumor seriously disease threaten human health and has become a serious social problem. Doxorubicin is a broad-spectrum anticancer drugwith good effect.Due to its serious side effects, such as cardiac toxicity,its clinical use is limited. Polyethyleneimine (PEI)-DNA-doxorubicin (DOX) complexes can effectively deliver doxorubicin to the target site and reduce its toxicity. In this experiment, we prepared a liver targeted nanoparticle with liposome-encapsulated PEI-DNA-DOX complexes in order to obtain lower toxicity, and hepatic targeting.Method:This experiment included three parts. First, determine the doxorubicin content determination conditions, preparation the PEI-DNA-DOX lipopolyplexes and investigating its pharmaceutical character. Next, inspecting the lipopolyplexes pharmacokinetic properties in vivo and establishing an analysis method in vivo and the dosage. Finally, examining the lipopolyplexes distribution and targeting evaluation in vivo.Result:1. The conditions of doxorubicin HPLC determination:mobile phase, methanol-0.01M phosphate buffer-glacial acetic acid (85:15:0.25; V/V); detector: fluorescence detector; detection wavelength, Ex:505nm, Em:559nm; flow velocity,1ml/min; column temperature,25℃; sample size,20μl.2. The best reaction conditions of DNA and DOX. Concentration of doxorubicin was60μg/ml in a buffer solution of pH=4; incubation60min in room temperature. Its N/P ratio was22, liposome/pDNA was210.3. Established analysis method of doxorubicin in vivo.the method has high specificity and good reproducibility. the standard curve of plasma samples: C=0.0458A-2.9078; R2=0.9997, the linear range was:0.375-192μg/ml. The results of doxorubicin solution pharmacokinetics:The process in vivo was accord with the three rooms model, t1/2pi=2.484min, t1/2a=27.778min, t1/2β=1042.892min. The process of LPD containing doxorubicin in vivo was also accord with the three rooms model, t1/2pi=8.244min, t1/2a=225.172min, t1/2β=269538.91min. According to the median lethal dose, final dosage of intravenous injection in mice was:0.6mg/kg.4. The results of tissue distribution show:compared with the doxorubicin solution, the distribution of doxorubicin lipopolyplexes in the heart was significantly reduced accumulation of doxorubicin was increased significantly in liver and spleen. Targeted evaluation showed that after intravenous injection LPD nanoparticles containing doxorubicin, the value of AUC in liver was3.6times greater than that in the other tissues. The Cmax of DOX-LPD polyplexes nanoparticles groups in the tissues were less than doxorubicin solution group, but in the liver and spleen tissue compared with doxorubicin solution was more than1.5times. The Re value of liver significantly greater than that in other tissues. The above three data indicated that the preparation of LPD-DOX lipopolyplexes has good liver targeting.Conclusion:Studied the doxorubicin-lipid-polycation-DNA polymers systematically, established determination of content analysis method of doxorubicin in vitro and in vivo, researched and evaluated thepreparation technology and prescription, encapsulation efficiency, stability, pharmacokinetics, tissue distribution and targeting of doxorubicin-lipid-polycation-DNA-polymerand. Preparation of LPD can significantly reduced cardiotoxicity of doxorubicin and improved its hepatic targeting.