【作者】 刘莎；

【导师】 薛玉英； 刘晶；

【作者基本信息】 东南大学 ， 公共卫生（专业学位）， 2018， 硕士

【摘要】 研究目的:兰索拉唑属于质子泵抑制剂(proton pump inhibitors,PPIs),可用于胃食管反流性疾病(gastroesophageal reflux disease,GERD)的治疗。右旋兰索拉唑是兰索拉唑的右旋对映体,而注射用右旋兰索拉唑则可用于口服疗法不适用的伴有出血的十二指肠溃疡。本研究遵照药物临床前毒性研究指导原则,采用啮齿类动物进行注射用右旋兰索拉唑急性毒性、亚急性毒性以及毒代动力学研究,以评价注射用右旋兰索拉唑的毒性效应,确定毒性靶器官,为注射用右旋兰索拉唑后续研究提供依据。同时与注射用兰索拉唑进行比较,为临床安全用药以及不良反应监控提供参考信息。研究方法:1.急性毒性研究:按照急性毒性试验设计原则,采用SD大鼠进行。注射用右旋兰索拉唑及注射用兰索拉唑设定剂量均依次为:250.0、225.0、202.5、182.3、164.0mg/kg,SD大鼠每组10只,雌雄各半,同时设溶媒对照组,按给药组给予同等体积的0.9%氯化钠注射液,采用尾静脉给药。给药后每天观察并记录动物毒性反应及死亡情况,连续14天,测定半数致死量(LD50)和95%可信区间。2.亚急性毒性研究:参考急性毒性研究结果制定SD大鼠的给药剂量。注射用右旋兰索拉唑高、中、低剂量依次为60、20、6mg/kg,另设注射用兰索拉唑(60mg/kg)和0.9%氯化钠注射液作为消旋和溶媒对照组。大鼠尾静脉注射给药,每天1次,连续4周。观察并记录动物的毒性反应和严重程度及停药后的发展和恢复情况。3.毒代动力学研究:80只SD大鼠分为注射用右旋兰索拉唑60、20、6mg/kg,注射用兰索拉唑(60mg/kg)和注射用左旋兰索拉唑组(60mg/kg),每组16只,雌雄各半。每天给药1次,连续4周。在给药后第1和28天时每组各取8只大鼠取血。取血点为给药前及给药后0、0.083、0.5、1、2、4、6h。用LC-MS/MS测定血浆中右旋兰索拉唑,左旋兰索拉唑的含量;运用DAS2.1.1程序计算毒代动力学参数。研究结果:1.急性毒性评价:注射用右旋兰索拉唑大鼠静脉给药半数致死量(LD50)为207.17mg/kg,95%可信区间为196.66--218.30mg/kg;对照药物注射用兰索拉唑大鼠静脉给药LD50为206.61mg/kg,95%可信区间为195.46--218.74mg/kg。观察期间动物一般状况良好、体重增长呈正常趋势,未发现大鼠出现明显中毒症状。解剖时肉眼观察主要脏器未见明显异常变化。2.亚急性毒性评价:注射用右旋兰索拉唑20mg/kg及以下剂量组大鼠在一般症状、体重、摄食量、血液生化、凝血时间、血清电解质方面与溶媒对照相比无显著性差异,病理检查给药4周后胸腺有轻度萎缩,但停药恢复4周后可以恢复正常。与溶媒对照组相比,60mg/kg剂量组的注射用右旋兰索拉唑以及同剂量的消旋对照药物EOS%,MCV有统计学差异,消旋对照组MCH有统计学差异;60mg/kg剂量组雄性大鼠在整个给药4周期间均于给药后出现短暂中枢神经系统毒性反应;雌性大鼠在前9天给药后出现于雄鼠相同的毒性反应,给药9天后四肢乏力,俯卧或者侧卧的现象消失,但雌鼠背部出现脱毛的现象,脱毛症状在恢复期结束前一周消失,雌鼠背部毛发恢复正常。连续给药4周后60mg/kg剂量组大鼠胸腺相对重量明显减轻并伴随病理改变,其发生率和严重程度与消旋对照组相当。恢复4周后,60mg/kg剂量组和消旋对照组胸腺均有所恢复。3.毒代动力学研究:根据血药浓度-时间数据结果显示几乎所有动物给药后5min血药浓度即可达峰值,其后血药浓度随时间迅速降低。使用DAS2.1.1软件计算毒代动力学参数发现,注射用右旋兰索拉唑6、20、60mg/kg各剂量组连续4周给药后雌性、雄性大鼠AUC(0-t)、AUC(0-∞)、Cmax均较1d首次给药的显著增加,说明连续给药4周后,注射用右旋兰索拉唑在雌性、雄性动物体内的暴露均增加。消旋对照组注射用兰索拉唑组连续给药4周给药后,雄性动物体内左/右旋兰索拉唑AUC(0-t)、AUC(0-∞)与1d首次的差异没有显著性,仅Cmax显著增高;雌性动物体内AUC(0-t)、AUC(0-∞)、Cmax则显著增加。相关性分析显示注射用右旋兰索拉唑对大鼠单次和多次给药体内毒代动力学参数均呈剂量线性关系。研究结论:注射用右旋兰索拉唑静脉给药4周对SD大鼠的相对安全剂量为20mg/kg/d及其以下剂量,其毒性作用靶器官主要为胸腺,大鼠急性和亚急性毒性反应程度与消旋对照品相当。注射用右旋兰索拉唑在大鼠体内呈现线性毒代动力学特征。本研究为注射用兰索拉唑临床毒副反应监测和安全用药提供了重要参考。更多还原

【Abstract】 Objective:Lansoprazole is a proton pump inhibitor(proton pump inhibitors,PPIs),which can be used in the treatment of gastroesophageal reflux disease(gastroesophageal reflux disease,GERD).Dexlansoprazole is the dextro-enantiomers of lansoprazole,and can be used for patients with duodenal ulcer with bleeding that are not suitable for oral therapy.This study followed the guidelines for preclinical toxicity studies of drugs to study the acute toxicity,subacute toxicity and pharmacokinetics of dexlansoprazole in rats.And to evaluate the toxicity of the dexlansoprazole for injection and find out the target organs.Thus to provide a theoretical basis for the follow-up studies.Also,a comparison study with lansoprazole for injection,a marketed drug,was performed to provides reference information for clinical safety medication and adverse reaction monitoring.Methods:1.Acute toxicity studyAccording to the principle of acute toxicity experimental design,the dosage for both dexlansoprazole for injection and lansoprazole for injection were 250.0,225.0,202.5,182.3,164.0 and 0 mg/kg respectively.The 10 SD rats(5 males and 5 females)in each group were intravenously administered.General behavior,adverse effects and mortality were recorded for up to 14 days post treatment.LD50 and 95%confidential intervals were measured.2.Subacute toxicity studyDexlansoprazole for injection was given intravenously at the dose of 60,20,6,and 0 mg/kg per day(n=20/group each sex)for 4 weeks to rats.Lansoprazole for injection with a dose of 60mg/kg were administered simultaneously.Clinical symptoms after treatment were observed.3.Toxicokinetics studyThe 80 SD rats(8 per group each sex)were randomly divided into five groups including three administration of different dose(60,20,6)mg/kg,injection with lansoprazole(60mg/kg)and injection with S-lansoprazole(60mg/kg).Blood samples of each animal were collected on the 1st and 28th day at before and 0,0.083,0.5,1,2,4 and 6 hours after the treatment.The concentration of the dexlansoprazole and lansoprazole in plasma was determined by LC-MS/MS.And the toxicokinetics parameters were calculated with DAS2.1.1 program.Results:1.Acute toxicityThe(LD50)of intravenous administration of dexlansoprazole and lansoprazole in rats is 207.17mg/kg(95%confidence interval 196.66--218.30 mg/kg)and 206.61mg/kg(95%confidence interval 195.46-218.74 mg/kg)respectively.During the 14-day study,the general condition of the animals was good,and the body weight gain showed not effected.No further signs of toxicity occurred,and no abnormality of organs noted in histopathological examinations.2.Subacute toxicityThere were no significant changes in body weight gain,food intake,biochemical parameters,coagulation time,serum electrolyte measurement and etc.in rats at dexlansoprazole 20mg/kg.And slight atrophy was found in thymuses but recoverable while treatment stopped at this dosage.EOS%and MCV changed significant in rats treated with both 60 mg/kg dexlansoprazole and lansoprazole.In the dexlansoprazole 60 mg/kg dose group,a sign of neurological symptom was observed in male rats right after administration.The same symptoms also found in female rats in the beginning 9 days after dosage,and which disappeared from the 10th day.But the female rats showed hair loss on the back then,and recovered one week advance in the end of recovery period.A weight loss of thymuses occurred in both 60 mg/kg dexlansoprazole and lansoprazole treated animal groups,and can be recovered to some extent in the recovery period.3.Toxicokinetics study:The results showed concentration in serum reached peak at 5 min post treatment,and decreased afterward rapidly.The AUC(0-t),AUC(0-∞)and Cmax of all dexlansoprazole dosage treated rats were observed significantly increased after 28-day administration compared to the single dose treatment.Suggested dexlansoprazole exposure ascension obviously after 4 weeks treatment in both male and female rats.Related analysis lead to a resulted that the toxicokinetic parameters has a linear relationship with dosages both in single and repeated dexlansoprazole treated rats.Conclusion:The relative safe dose of intravenous administration for 4 weeks of the dexlansoprazole for injection was not more than 20 mg/kg/day in rats,and thymus is the main target organ.The toxicity in both acute and subacute studies with dexlansoprazole and lansoprazole are similar.Characteristics of linear toxicokinetics of dexlansoprazole in female and male rats were discovered.This study provided important information for the monitoring toxicity and adverse effects therefore to applicate safely clinically of the dexlansoprazole for injection.更多还原