Preparation Of Podophyllotoxin Liposome Chitosan Film And Its Researches On Animals

Objective:1. Prepare the podophyllotoxin liposomes liquid and observe its diameter,embedding ratio and stability, etc, and discuss the relation among them.2. Prepare the chitosan film and observe the stability to podophyllotoxinliposomes.3. Observe the suscitation of podophyllotoxin liposomes chitosan film torabbits' vagina.4. Study the systemic absorption after topical applying podophyllotoxinliposomes chitosan film to rabbits' cervix.5. Set up the method of detecting podophyllotoxin in rabbits' blood byHPLC.Methods:1. The podophyllotoxin liposomes liquid was prepared by rotary filmevaporation and ultrasonic wave. The embedding ratio of podophyllotoxinwas determined by HPLC. Film agent were made of chitosan and 3% Aceticacid (6g: 100 ml). The podophyllotoxin liposomes chitosan film and thepodophyllotoxin tincture were contrasted in the suscitation experiment.2. 40 rabbits were divided into four groups randomly, which including onetest group and three control groups, each group ten rabbits. The test rabbitswere applied with podophyllotoxin liposomes chitosan film and the controlrabbits were applied with podophyllotoxin tincture, podophyllotoxinointment and podophyllotoxin chitosan film respectively. Rabbits' blood wasdrew out in regular time and was extracted by dichlormethane and wasdetermined by HPLC and the data was analysed by spss10.0.Result:1. The podophyllotoxin liposomes prepared by rotary film evaporation andultrasonic wave are all unilaminar vesicles. The diameter is from 0.2 μM to1.0μ m and the embedding ratio is between 70% to 80% and the averageembedding ratio is 76.3%. The podophyllotoxin liposomes chitosan filmunder the room temperature didn't appear obvious variety in six months.2. The suscitation of 0.5% podophyllotoxin liposomes chitosan film torabbits' vagina is lighter than that of 0.5% podophyllotoxin tincture and thefilm agent is good adherent to rabbits' cervix.3. The podophyllotoxin concentration of test group is always lower than thatof three control groups, the highest podophyllotoxin concentration value oftest group is 175±43.957ng/ml and that of control groups is respective 893.8±52.548 ng/ml, 499.4±44.1694 ng/ml and 458.6±45.3338ng/ml ( p<0.001).The AUC of podophyllotoxin of the control groups are respective 4.83-fold ,2.63-fold and 2.58-fold higher than that of the test group.Conclusion:1. The stability of podophyllotoxin liposomes was improved by the chitosanfilm and the suscitation to rabbits' vagina is lighter than that of thepodophyllotoxin tincture and is good fit with rabbits' cervix.2. The systemic absorption of podophyllotoxin of the test group is lower thanthat of the control groups, It is sure that the side effect of podophyllotoxincan be decreased by liposomes agent.3. The HPLC method is precise, facile and reliable, and it can be used withdetermining the podophyllotoxin concentration of animals' blood.