贝伐单抗膜联蛋白A5脂质体兔眼点眼后眼内组织药物浓度变化和药代动力学特征

发布时间：2018-04-25 20:06

本文选题：贝伐单抗 + 膜联蛋白A5脂质体　；参考：《新乡医学院》2017年硕士论文

【摘要】：背景目前,新生血管性眼病的发病率不断上升。贝伐单抗是一种治疗新生血管性眼病的单克隆抗体药物。由于各种眼部屏障的存在,局部点眼治疗无法在眼内到达有效浓度。目前临床上采用玻璃体注射此类药物来治疗眼内新生血管性疾病。但多次玻璃体注射可能产生一些严重并发症。近来研究发现,采用膜联蛋白A5脂质体作为载体可以使贝伐单抗穿过角膜的能力有较大的提高,具有成为对眼内新生血管性疾病进行无创性治疗方法的潜力。因此,对贝伐单抗膜联蛋白A5脂质体滴眼液在兔眼的药物代谢动力学进行研究将会对临床治疗眼内新生血管性疾病能够提供帮助。目的为减少眼部并发症,提高药物局部应用的眼内通透性,本研究通过制备一种新型贝伐单抗膜联蛋白A5脂质体滴眼液,通过对实验动物进行局部点眼,对其眼部药动学进行研究,为临床治疗视网膜脉络新生血管性疾病提供实验基础与理论依据。方法1.制备药品:贝伐单抗来自购买的成品。贝伐单抗脂质体由脂质体微泡和贝伐单抗采用双水化法制成。贝伐单抗膜联蛋白A5脂质体由贝伐单抗脂质体加酶联蛋白A5于搅拌器上低速旋转制成。2.实验动物分组:105只健康无眼病的新西兰大白兔,不分雌雄,随机数字表法分为实验组、对照组1、对照组2,每组35只,每组再随机分为7个亚组,每个亚组5只,分别对应给药后的7个时间点(5min、15min、30min、1h、2h、4h、8h)。3.给药及样品采集:实验组新西兰大白兔单眼接受贝伐单抗膜联蛋白A5脂质体局部点眼50μL;对照组1新西兰大白兔单眼接受贝伐单抗脂质体局部点眼50μL;对照组2新西兰大白兔单眼接受贝伐单抗局部点眼50μL。各组分别在给药后预定时间点处死动物,摘取眼球,取房水及玻璃体、视网膜脉络膜标本,备用。4.获取药代动力学参数及统计学分析:酶联免疫吸附法计算各样本浓度值。应用DAS 2.1.1软件计算各个药物代谢动力学参数,拟合曲线。应用SPSS 19.0软件进行统计分析,采用方差分析和LSD检验对数据进行统计学分析。结果1.各组实验数据证明贝伐单抗膜联蛋白A5脂质体在各个已测时间点的浓度明显高于对照组1和对照组2(P0.05)。2.实验组视网膜脉络膜组织中的贝伐单抗浓度高于体外贝伐单抗对hVEGF的IC50。结论贝伐单抗膜联蛋白A5脂质体有望成为治疗新生血管性眼病的新剂型,通过局部点眼的方式给药从而避免眼内注射,减少并发症。
[Abstract]:Background at present, the incidence of neovascular ophthalmopathy is increasing. Bevacizumab is a monoclonal antibody drug for the treatment of neovascular ophthalmopathy. Due to the presence of various eye barriers, local eye therapy cannot reach the effective concentration in the eye. At present, vitreous injection is used to treat intraocular neovascularization. But multiple vitreous injections can lead to serious complications. Recently, it has been found that the ability of bevacizumab to penetrate the cornea can be improved by using A5 liposome as carrier, and it has the potential to be a non-invasive therapy for intraocular neovascularization. Therefore, the study of pharmacokinetics of bevacizumab A5 liposome eye drops in rabbit eyes will be helpful for the clinical treatment of intraocular neovascularization. Objective to reduce the ocular complications and improve the intraocular permeability of the drug, we prepared a new type of bevacizumab membrane binding protein A 5 liposome eye drops, and made a local eye spot on the experimental animals by the preparation of a new type of bevacizumab A5 liposome eye drops. The study of ocular pharmacokinetics provides experimental and theoretical basis for the clinical treatment of retinal choroidal neovascularization. Method 1. Preparation of drugs: bevacizumab comes from the finished product purchased. The bevacizumab liposome was prepared by double hydration of liposome microbubbles and bevacizumab. The bevacizumab membrane linked protein A5 liposome was prepared from bevacizumab liposome and enzyme linked protein A5 by rotating at low speed on the agitator. The experimental animals were divided into 10 groups: 105 healthy New Zealand white rabbits without eye disease, male and female, randomly divided into experimental group, control group 1, control group 2, each group 35. Each group was randomly divided into 7 subgroups, 5 in each subgroup. The results showed that 7 time points after administration were 5 min ~ 15 min ~ 30 min ~ (-1) ~ 1 h ~ (-1) ~ 2 h ~ (-1) ~ 4 h ~ (8) h ~ (-1) ~ 8 h ~ (-1) ~ (3). Administration and sample collection: the experimental group New Zealand white rabbits received bevacizumab A5 liposome 50 渭 L locally; the control group 1 New Zealand white rabbit single eye received bevacizumab liposome 50 渭 L; control group 2 The local point of bevacizumab was 50 渭 L in single eye of blue white rabbit. The animals in each group were killed at a predetermined time point after administration, eyeball was removed, aqueous humor and vitreous body, retinal choroid specimen were taken, reserve. 4. The pharmacokinetic parameters and statistical analysis were obtained: enzyme linked immunosorbent assay (Elisa) was used to calculate the concentration of each sample. DAS 2.1.1 software was used to calculate the pharmacokinetic parameters and fit the curve. SPSS 19.0 software was used for statistical analysis, and ANOVA and LSD test were used for statistical analysis. Result 1. The experimental data showed that the concentration of bevacizumab A5 liposome at each time point was significantly higher than that of control group 1 and control group 2 (P0.05 路2). The concentration of bevacizumab in retinal choroid tissue of experimental group was higher than that of in vitro bevacizumab on hVEGF. Conclusion the bevacizumab A5 liposome is expected to be a new dosage form for the treatment of neovascular ophthalmopathy, which can avoid intraocular injection and reduce complications.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R77

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