基于Caco-2细胞模型的乙酰半胱氨酸渗透性评价研究

Evaluation of acetylcysteine permeability based on Caco-2 cell model

目的检测乙酰半胱氨酸的表观渗透系数(Papp)值,为其生物药剂学分类系统(BCS)分类评价提供重要参考依据。方法构建致密性良好的Caco-2单层细胞模型,分析比较地高辛等外排机制药物、茶碱等高渗透性药物、二甲双胍等中渗透性药物、氯噻嗪等低渗透性药物、荧光黄作为零渗透性药物的Papp值,以HPLC-UV法分别检测并鉴定目标药物3 mmol·L^(-1)、300μmol·L^(-1)和30μmol·L^(-1)乙酰半胱氨酸的肠道渗透性。结果连续21 d的电阻值检测显示本细胞模型致密性良好,地高辛等外排机制药物在此模型上均呈现为良好的外排功能。在本项研究的高渗透性药物中,普萘洛尔的Papp值最大,米诺地尔的Papp值最小,中渗透性药物雷尼替丁的Papp值最大,低渗透性药物氯噻嗪、甘露醇、PEG 400、阿昔洛韦、纳多洛尔的Papp值与高渗透性药物相差较大,但与中渗透性药物相差较小;零渗透性药物荧光黄Papp值较小。而受试药物乙酰半胱氨酸Papp_((A-B))值>米诺地尔的Papp_((A-B))值6.43×10^(-6)cm·s^(-1)(CV,6.99%),差异有统计学差异意义(P<0.05)。结论本研究在指导原则中选取的5种高渗透性工具药物中,检测得到米诺地尔的Papp_((A-B))值最小。因此以米诺地尔为高渗透性药物的内标,即可认为目标药物乙酰半胱氨酸是高渗透性药物。对其一致性评价中BCS分类的判断即可认为乙酰半胱氨酸属于BCS I类药物。

Objective To investigate the intestinal permeability of different biopharmaceutical classification system(BCS)tool drugs in the Bioequivalence Exemption Guidelines issued by the State Food and Drug Administration,we have evaluated the apparent permeability coefficient of acetylcysteine,thus providing an important reference for its BCS classification.Methods Caco-2 cell model was used to analyze and compare theophylline et al.(high permeability drugs),metformin et al.(medium permeability drugs),chlorothiazide et al.(low permeability drugs),fluorescent yellow(zero permeability drugs)and digoxin et al.(efflux mechanism drugs).Then intestinal permeabilities of 3 mmol·L^(-1),300μmol·L^(-1)and 30μmol·L^(-1)acetylcysteine were detected and identified by HPLC-UV method.Results The resistance value detection of Caco-2 cell model for consecutive 21 days showed that the cell model was compact.Digoxin et al.all showed good efflux.By detecting the high permeability drug propranolol,we have determined the maximum value of the apparent permeability coefficient,and Papp of ranitidine was in the highest value among all medium permeability drugs.For low permeability drug mannitol,PEG 400,acyclovir,their Papp values were much lower than those of high permeability drugs,but smaller differences with medium permeability drugs.Papp value of zero permeability drug fluorescence yellow is the lowest.The Papp_((A-B)) value of acetylcysteine was higher than that of minoxidil 6.43×10^(-6)cm·s^(-1)(CV,6.99%)minoxidil of the internal standard for hyperpermeable drug.Conclusion In this study,the Papp_((A-B)) value of minoxidil was the lowest among the five high-permeability tool drugs selected in the guidelines,which was lower than the tested drug acetylcysteine.Therefore,the tested drug was considered to be hyperpermeable.According to the BCS classification in the consistency evaluation,acetylcysteine could be considered as a BCS class I drug.