The Preparation, Evaluation And Anti-tumor Activity Of The Irinotecan Long-circulating Nano-liposome

Purpose:The purpose of this study was to explore the preparation process, evaluating method and anti-tumor activity of ILCNL and to lay a foundation of further study of the preparation.Methods:(1) Orthogonal design was used to optimal the preparation method The different kinds of preparation methods were utilized to investigate and finalize the best preparation method. At last, the ethanol injection-ammonium sulfate gradient method was obtained to prepare Irinotecan long-circulating nano-liposome. Then the best prescription was tested by the orthogonal experiment.(2) Quality evaluation of ILCNLAccording to the best prescription, Irinotecan long-circulating nano-liposome was produced and examined on its particle size, zeta potential, shape, entrapment efficient, in vitro release, stability and so on.(3) Anti-tumor activity of ILCNLThe effect of Irinotecan long-circulating nano-liposome on three kinds of human rectal carcinoma cancer cells were tested by MTT method. At the same time, the effect of ILCNL on the cell morphology of HepG2was investigated.Results:(1) Preparation of the ILCNLThe ethanol injection-ammonium sulfate gradient method was selected to prepare this kind of liposome. The results showed good characteristics of size distribution (110-120nm), entrapment efficiency (92.67%) and drug loading (9%). The best preparation was:phospholipid300mg,10mg/mL, cholesterol50mg, PEG200050mg. The volume ratio of ammonium sulfate and the ethanol was3:1. The mass ratio of lecithin and irinotecan was10:1, and the mass ratio of lecithin and PEG2000was5:1. Water bath temperature was40℃.(2) Quality evaluation of ILCNLAccording to the best prescription, the average size of ILCNL was about115nm, zeta potential was-14.7mV, PH value was6.474, the entrapment efficiency was92.67%, the liposome was more stable at room temperature, and the drug release of ILCNL was slow?.(3) Anti-tomor activity of irinotecan loaded long-circulating nano-liposomeCompared with the CPT-11, the ILCNL was more effective to inhibit HCT-15, lovo and colo320cells and showed a concentration and time dependent manner in vitro. While at the same concentration, ILCNL had more potent killing function than that of free CPT-11. According to AO/EB method, ILCNL could induce changes in morphology of HepG2cells and cause their apoptosis.Conclusions:The preparation process of ILCNL was optimized by orthogonal design, and an effective quality evaluation method was created. This kind of liposome could protect the structure of lactone ring, prolong the drug action time, and enhance the anti-tumor activity. The study accumulated more data on liposome research for further clinic exploration.