三苯双脒抗华支睾吸虫药效与机制的研究

发布时间：2018-05-03 04:19

本文选题：三苯双脒 + 华支睾吸虫　；参考：《中国疾病预防控制中心》2013年博士论文

【摘要】：根据第二次全国人体寄生虫病分布调查的估算,我国的华支睾吸虫感染人数已达1249万。由于缺乏预防食源性吸虫病的疫苗和人们的饮食方式难以改变,药物化疗仍然是控制该类疾病的主要手段。目前用于治疗华支睾吸虫感染的药物主要是吡喹酮,根据世界卫生组织的推荐,吡喹酮用于治疗华支睾吸虫感染的适宜剂量疗程是每次口服25mg/kg,3次/d,连给2d。该疗法虽有较好的疗效,但应用于大规模群体治疗具有一定的难度,因此,继续寻求新的抗华支睾吸虫病药物仍是十分必要的。近年来动物试验发现顿服单剂三苯双脒对华支睾吸虫和麝猫后睾吸虫有很好的杀灭效果,且其对麝猫后睾吸虫的疗效在临床试验中得到了验证。本论文先通过实验室研究确定三苯双脒对华支睾吸虫的疗效,并初步探讨其抗虫机制及在胆汁中的代谢动力学,然后尝试通过三苯双脒治疗肠道线虫和华支睾吸虫混合感染的病人,评价三苯双脒对感染华支睾吸虫的疗效及安全性,为华支睾吸虫病的现场群体化疗提供参考。在实验室研究中,我们首先通过观察不同剂量的三苯双脒对体外培养的华支睾吸虫和对感染华支睾吸虫囊蚴的仓鼠的疗效,评价其对体外和体内的华支睾吸虫的作用。该研究明确了三苯双脒对体外培养的华支睾吸虫具有很强的杀灭作用,且对其最低致死剂量仅为吡喹酮的一半；对仓鼠体内的华支睾吸虫童虫和成虫均有好的疗效,其中100mg/kg三苯双脒治疗华支睾吸虫成虫可有高于90%的减虫率,即使剂量低至12.5mg/kg仍可有50%的减虫率,效价强度显著高于吡喹酮。其次,我们通过观察三苯双脒对华支睾吸虫超微结构的影响,对其抗虫机制进行了初步探讨。大鼠经胃灌服三苯双脒300mg/kg后4h、8h、24h、48h和72h分别剖杀取虫,经预处理后置于透射电镜下观察。结果显示三苯双脒能引起华支睾吸虫皮层乳突的肿胀、破溃和溶解,层皮内杆状颗粒、盘状颗粒和膜样小泡的减少,线粒体和内质网的减少,线粒体形成空泡以及皮层细胞核膜损伤。再次,本论文对三苯双脒代谢物氨脒在大鼠体内的药物代谢动力学进行了研究。首先建立了大鼠血浆和胆汁中氨脒浓度的测定方法,然后用200mg/kg三苯双脒治疗大鼠后于不同时间点取血样及收集胆汁,同样时间点收集未经治疗的空白大鼠的血样和胆汁,经处理后进样测定,绘制血药浓度—时间曲线、胆汁药物浓度—时间曲线以及胆汁排泄速率和累计排泄量曲线。结果发现氨脒在血浆中的达峰时间Tmax为0.90±0.22h,峰浓度Cmax为8.12±2.05mg/L.而氨脒在胆汁中的达峰时间Tmax为1.15±0.42h,峰浓度Cmax为11.16±2.07mg/L,11h内氨脒的胆汁累积排泄量为40.12±12.00μ g,累积排泄率0.38‰,给药后0.5～3h的胆汁排泄速率最大。最后,本论文还通过观察三苯双脒治疗肠道线虫和华支睾吸虫混合感染的病人的治愈率、虫卵减少率和不良反应,评价三苯双脒抗华支睾吸虫疗效和安全性。通过伦理审查后,在试验地区采用Kato-Katz法进行基线调查,根据入选标准和排除标准确定拟服药人员,随机分为3组,签订知情同意书及经过临床医生体检后,分别口服吡喹酮(总剂量为75mg/kg)、三苯双脒(400mg,顿服)、三苯双脒(200mg*2次)。部分接受三苯双脒治疗的病人在治疗前和治疗后24h检测血尿常规、心电图和肝肾功能(空腹抽血)。研究人员对服药人员治疗后48h内进行随访,询问有无不良反应并记录具体情况。治疗后3w,收集病人粪样,用Kato-Katz法检测并计数粪样的华支睾吸虫、钩虫、蛔虫、鞭虫和绦虫的虫卵数,计算华支睾吸虫的治愈率和虫卵减少率。然后根据第一次试验结果,邀请未治愈病人,按照同样的操作标准进行第二次相同药物治疗,并评价疗效与安全性。最后所有未治愈的受试者,采用吡喹酮标准疗法进行治疗。本研究发现顿服400mg的三苯双脒组的治愈率和虫卵减少率与吡喹酮组无显著性差异,但不良反应较吡喹酮组少,且吡喹酮组更易出现眩晕的不良反应。综合以上各部分的研究结果,我们认为三苯双脒对华支睾吸虫具有良好的治疗作用,但是还需开展大规模多中心的临床试验以进一步验证其疗效和评价安全性,为华支睾吸虫病现场群体化疗提供依据。
[Abstract]:According to the second national survey of the distribution of human parasitology, the number of Clonorchis sinensis infection in China has reached 12 million 490 thousand. Due to the lack of vaccines to prevent foodborne parasitition and the difficult change of people's diet, drug and chemotherapy is still the main means to control the disease. If praziquantel, according to the recommendation of the WHO, the suitable dose of praziquantel for the treatment of Clonorchis sinensis infection is 25mg/kg every time, 3 times /d, even to 2D., although it has a good effect, but it is difficult to apply to mass group therapy, because of this, continuing to seek new anticlonorchiasis drugs is still ten. In recent years, the animal test found that the single dose of three benzamine had a good killing effect on Clonorchis sinensis and the posterior testosterone of the cat cat, and its effect on the posterior testosterone of the cat was verified in clinical trials. First, the effect of three benzamid on Clonorchis sinensis was determined by laboratory study, and its resistance was preliminarily discussed. The mechanism of the insect and the metabolic kinetics in the bile, and then try to evaluate the efficacy and safety of the three benzamid in the infection of Clonorchis sinensis through the treatment of the mixed infection of intestinal nematodes and Clonorchis sinensis by three benzamines, and provide reference for the field group chemotherapy of Clonorchis sinensis.
In laboratory studies, we first evaluated the effect of different doses of three benzamid on Clonorchis sinensis in vitro and hamsters infected with cysts of Clonorchis sinensis in vitro, and evaluated its effect on Clonorchis sinensis in vitro and in vivo. The study clearly showed that three benzamiprid was strongly killed in vitro culture of Clonorchis sinensis. The minimum lethal dose was only half of the praziquantel, which had a good effect on the adults and adults of Clonorchis sinensis in the hamster, and 100mg/kg three benzamiprid could have a higher than 90% worm reduction rate in the treatment of Clonorchis sinensis adults. Even if the dose was low to 12.5mg/kg, the rate of insect reduction was 50%, and the titer intensity was significantly higher than that of praziquantel.
Secondly, by observing the ultrastructure of three benzamidin on the ultrastructure of Clonorchis sinensis, the mechanism of its insect resistance was preliminarily discussed. The rats were killed by 4h, 8h, 24h, 48h and 72h after the gastric perfusion of 300mg/kg, and then observed under transmission electron microscope after preprocessing. The results showed that three benzamidamidin could cause the clonorchis sinensis cortical mastoid process. Swelling, ulceration and dissolution, rod like particles in the skin, disc and membrane like vesicles decreased, mitochondria and endoplasmic reticulum decreased, mitochondria formed vacuoles, and cortical cell nuclear membrane damage.
In this paper, the pharmacokinetics of amamamid, a three benzamid metabolite, was studied in rats. First, the determination of amamamid concentration in rat plasma and bile was established, and then 200mg/kg three benzamiprid was used to collect blood samples at different time points and collect bile juice at different time points, and the untreated blank was collected at the same time. The blood samples and bile of rats were measured after treatment. The concentration of blood drug concentration time curve, the concentration time curve of bile drugs and the curve of bile excretion rate and cumulative excretion were drawn. The results showed that the peak time of amamamid in plasma was Tmax 0.90 + 0.22h, the peak concentration Cmax was 8.12 + 2.05mg/L. and the peak time of amamiamid in the bile was Tm The ax was 1.15 + 0.42h, the peak concentration Cmax was 11.16 + 2.07mg/L, the accumulated excretion of bile in 11h was 40.12 + 12 mu g, and the cumulative excretion rate was 0.38 per thousand. The rate of bile excretion in 0.5 ~ 3H after administration was the largest.
Finally, the efficacy and safety of three benzamid in the treatment of mixed infection of intestinal nematodes and Clonorchis sinensis, the rate of egg reduction and adverse reactions, and the efficacy and safety of the three benzamid in the anti Clonorchis Clonorchis were also evaluated. After the ethical review, the Kato-Katz method was used in the baseline survey in the experimental area. The people were divided into 3 groups randomly, including 3 groups. After signing informed consent and after medical examination, they were taken oral praziquantel (total dose of 75mg/kg), three benzendine (400mg, meal), and three benzenamiprid (200mg*2 times). The patients receiving three benzendine treated and treated with three benzamines were tested for hematuria routine, electrocardiogram and liver and kidney before and after treatment. Function (fasting blood). The researchers were followed up in 48h after treatment, asked for adverse reactions and recorded specific conditions. After treatment, 3W, collected patient feces, detected and counted feces of Clonorchis sinensis, hookworm, Ascaris, flagellate and tapeworm eggs, and calculated the cure rate of Clonorchis sinensis and the decrease of eggs by Kato-Katz. Rate. Then according to the results of the first trial, the uncured patients were treated with the same medication for second times of the same drug treatment, and the efficacy and safety were evaluated. Finally, all uncured subjects were treated with praziquantel standard therapy. This study found the cure rate and the rate of egg reduction in the three benzamid group of 400mg. There was no significant difference between the praziquantel group and the praziquantel group, but the adverse reaction was less than that in the praziquantel group, and the praziquantel group was more likely to have adverse reactions to vertigo.
We think that three benzamiprid has a good therapeutic effect on Clonorchis sinensis, but we need to carry out large-scale multicenter clinical trials to further verify its efficacy and evaluate safety, and provide a basis for the field group chemotherapy of Clonorchis sinensis.

【学位授予单位】：中国疾病预防控制中心
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R532.23;R969

【参考文献】