体外Caco-2细胞模型在药物吸收中的应用进展

目的:介绍Caco-2细胞模型的特征及其在药物吸收中的应用.方法:分析国内外近期相关文献,对Caco-2细胞模型在药物吸收动力学中的研究进行概述.结果:Caco-2细胞模型用于药物动力学研究,可预测药物在体内的吸收和代谢.结论:体外Caco-2细胞模型在药物吸收过程的研究中有重要意义.     

Caco-2细胞模型在药物吸收研究中的应用

Caco-2细胞(thehumancoloncarcino-maline)模型是最近十几年来国外广泛采用的一种研究药物肠吸收的体外模型,它具有如下优点:与动物试验相比,培养细胞要比培养动物更省时更经济;可测定药物的细胞摄取及胯膜转运;Caco-2细胞内有药物代谢酶,可在有代谢状况下测定药物的胯膜转运;Caco-2细胞易于培养且生命力强;Caco-2细胞来源是人结肠癌细胞,同源性好;可用于区分肠腔内不同吸收途径的差别。1.Caco-2细胞模型建立Caco-2细胞适宜在37℃、含5%CO2的环境中培养,采用DMEM培养基,且在培养基中应含有10%胎牛血清、1%非必需氨基酸、1%谷氨酰胺和青…     

Caco-2细胞模型在药物研究中的应用

Caco2细胞模型广泛用于药物的吸收、代谢以及毒性研究,Caco2细胞模型作为药物吸收研究的一种快速筛选工具,在抗癌药物、无机药物、中药的研究方面得到了广泛的应用,成为药物研究的重要手段。     

Caco-2细胞模型在药物开发中的应用

从20世纪80年代开始，国外开始应用一种人结肠癌细胞(the human colon carcinoma cell line，Caco-2细胞)体外培养模型进行药物吸收的研究，目前该模型正被广泛用于药物开发中。caco-2细胞可在培养条件下自发进行上皮样分化并可以形成紧密联结，分化出绒毛面AP(apical，肠腔侧)和基底面BL(basolateral，肠壁侧)，其形态学、标志酶的功能表达、     

隐丹参酮在Caco-2细胞模型中的吸收机制

目的 研究隐丹参酮在Caco-2细胞模型中的吸收机制。方法 用Caco-2细胞单层模型研究隐丹参酮的双向转运，并考察时间、药物浓度及抑制剂对隐丹参酮吸收的影响。用高效液相色谱法检测药物浓度，计算其表观渗透系数。结果 隐丹参酮在Caco-2细胞模型中，从单层细胞层顶端到基底端的转运大于基底端到顶端的转运，随时间和浓度的增加，药物吸收呈饱和趋势，且可被其结构类似物丹参酮Ⅱ。竞争性抑制。结论 隐丹参酮在Caco-2细胞模型中的吸收主要是由载体介导的主动转运，且该主动转运的载体位于Caco-2细胞单层的顶端。     

抗肿瘤药物西美替尼在Caco-2细胞模型中的吸收转运

目的研究抗肿瘤药物西美替尼在Caco-2细胞模型中的吸收转运。方法建立Caco-2细胞转运模型,采用HPLC法测定药物浓度,计算表观渗透系数(P_app),研究西美替尼的Caco-2细胞跨膜转运情况。结果西美替尼在转运过程中没有明显的浓度依赖性。在低浓度范围内,药物的转运速率随着浓度的增加而增加;在较高浓度时达到饱和。西美替尼不同浓度的P_app值(3.91×10^-5、3.29×10^-5、1.90×10^-5和0.95×10^-5cm·s^-1)基本都大于难吸收药物的临界值(1×10^-5cm·s^-1)。西美替尼在Caco-2细胞中的转运呈现较强的方向性,从肠腔面到基底面的P_app值显著大于从基底面到肠腔面的P_app值（2.36～7.58倍）。ATP抑制剂叠氮化钠能显著降低西美替尼的正向转运程度,并升高其反向转运程度。当加入葡萄糖后,从肠腔面到基底面的跨膜转运程度显著降低。结论西美替尼主要是以转运载体介导的主动转运方式被吸收,其小肠吸收情况较好。     

Caco-2细胞模型在天然药物吸收研究中的应用

目的Caco-2细胞模型为经典的口服药物体外吸收模型。此文介绍了Caco-2细胞模型的来源与特点,综述了Caco-2细胞模型在天然药物吸收研究中的应用现状,并对Caco-2细胞模型在天然药物研究中的应用前景进行了探讨。     

Caco-2细胞模型在药物动力学研究中的应用

药物动力学研究中 ,涵盖着吸收、分布、生物转化和排泄四个过程。口服给药是临床最常见的途径 ,药物发挥作用 ,关键在于吸收。吸收过程和排泄过程的研究对药物动力学而言至关重要。近年来 ,通过肠上皮细胞的培养来作为研究药物吸收、代谢机制的体外模型取得令人鼓舞的进展。最具有代表性的细胞是来自结肠癌细胞系的Caco 2细胞 (thehumancolonadenocarcinomacelllines的简称 )。研究发现了药物透过Caco 2单细胞层的体外过程与药物口服后在肠中的吸收和代谢有良好的相关性 ,使得Caco 2细胞成为研究药物吸收和代谢的最经典的体外细胞模型[1,2…     

矢车菊黄素在Caco-2细胞模型中的吸收机制研究

目的 研究矢车菊黄素（centaureidin）在Caco-2细胞单层模型中的吸收机制。方法 以HPLC分析矢车菊黄素浓度,用Caco-2 细胞单层模型评价吸收时间、药物浓度、介质pH值、抑制剂等对矢车菊黄素吸收的影响,研究矢车菊黄素的吸收机制,计算表观渗透系数（apparent permeability coefficient, Papp）。结果 药物的吸收与药物浓度和吸收时间正相关;弱酸性介质条件下有利于药物的吸收;2,4-二硝基酚（DNP）对药物吸收无影响,但异博定（verapamil）可增加药物的吸收;从肠腔侧到基底侧的转运小于基底侧到肠腔侧的转运。结论 矢车菊黄素在Caco-2细胞模型中的吸收主要是被动转运,受P-糖蛋白的外排作用。     

Caco-2细胞模型——药物吸收研究的有效“工具”

吸收过程是决定口服药物生物利用度的重要因素 ,然而很多药物的吸收机制还不明确。Caco 2细胞模型是目前最好的体外吸收模型 ,在药物的吸收过程及吸收机制的研究方面有广泛的应用 ,尤其在中药吸收研究方面 ,Caco 2细胞模型的应用成为目前的热点。另外 ,Caco 2细胞模型在药物代谢方面也有应用。因此 ,Caco 2细胞模型将成为药物吸收研究的重要手段 ,有助于加快新药筛选和开发的速度。     

利用Caco-2细胞模型研究白鲜碱和茵芋碱在人小肠的吸收

目的:研究中药化学成分白鲜碱和茵芋碱的人小肠吸收情况。方法:利用人源结肠腺癌细胞系Caco-2细胞单层模型观察白鲜碱和茵芋碱由绒毛面(AP端)到基底面(BL端)、BL端到AP端2个方向的转运过程。应用偶联紫外检测器的高效液相色谱法对上述2种生物碱进行定量分析,计算转运参数和表观渗透系数,并与阳性对照药普萘洛尔和阿替洛尔进行比较。结果:由AP端到BL端,白鲜碱和茵芋碱的表观渗透系数(Papp)分别为(1.59±0.14)×10-5cm.s-1和(3.19±0.09)×10-5cm.s-1;由BL端到AP端,白鲜碱和茵芋碱的Papp分别为(2.57±0.33)×10-5cm.s-1和(5.86±0.49)×10-5cm.s-1,与在Caco-2单层细胞模型上呈良好吸收的阳性对照药普萘洛尔的基本一致。结论:白鲜碱和茵芋碱可以通过小肠上皮细胞被动吸收进入体内,属于吸收良好的化合物。     

Involvement of intestinal permeability in the oral absorption of clarithromycin and telithromycin

The involvement of intestinal permeability in the oral absorption of clarithromycin (CAM), a macrolide antibiotic, and telithromycin (TEL), a ketolide antibiotic, in the presence of efflux transporters was examined. In order independently to examine the intestinal and hepatic availability, CAM and TEL (10 mg/kg) were administered orally, intraportally and intravenously to rats. The intestinal and hepatic availability was calculated from the area under the plasma concentration–time curve (AUC) after administration of CAM and TEL via different routes. The intestinal availabilities of CAM and TEL were lower than their hepatic availabilities. The intestinal availability after oral administration of CAM and TEL increased by 1.3‐ and 1.6‐fold, respectively, after concomitant oral administration of verapamil as a P‐glycoprotein (P‐gp) inhibitor. Further, an in vitro transport experiment was performed using Caco‐2 cell monolayers as a model of intestinal epithelial cells. The apical‐to‐basolateral transport of CAM and TEL through the Caco‐2 cell monolayers was lower than their basolateral‐to‐apical transport. Verapamil and bromosulfophthalein as a multidrug resistance‐associated proteins (MRPs) inhibitor significantly increased the apical‐to‐basolateral transport of CAM and TEL. Thus, the results suggest that oral absorption of CAM and TEL is dependent on intestinal permeability that may be limited by P‐gp and MRPs on the intestinal epithelial cells. Copyright © 2014 John Wiley & Sons, Ltd.     

Enhancement of paracellular transport of heparin disaccharide across Caco-2 cell monolayers

The enhancement of paracellular transport of heparin disaccharide using several absorption enhancers across Caco-2 cell monolayers was tested. The cytotoxicity of these enhancers was also examined. The enhancing effects by Quillaja saponin, dipotassium glycyrrhizinate, 18beta-glycyrrhetinic acid, sodium caprate and taurine were determined by changes in transepithelial electrical resistance (TEER) and the amount of heparin disaccharide transported across Caco-2 cell monolayers. Among the absorption enhancers, 18beta-glycyrrhetinic acid and taurine decreased TEER and increased the permeability of heparin disaccharide in a dose-dependent and time-dependent manner with little or negligible cytotoxicity. Our results indicate that these absorption enhancers can widen the tight junction, which is a dominant paracellular absorption route of hydrophilic compounds. It is highly possible that these absorption enhancers can be applied as pharmaceutical excipients to improve the transport of macromolecules and hydrophilic drugs having difficulty in permeability across the intestinal epithelium.     

Apparent active transport of MDMA is not mediated by P-glycoprotein: a comparison with MDCK and Caco-2 monolayers

Amphetamines and their methylenedioxy derivatives generically display similar behavioral, physiologic and toxic effects. Inconsistent pharmacokinetic and toxicity data for methylenedioxymethamphetamine (MDMA) may suggest that active drug transporters are interacting with these compounds, and thus altering drug absorption and tissue distribution. In vitro models of CNS accumulation and intestinal drug transport were used to assess efflux transport of MDMA. Madin-Darby kidney cell epithelial (MDCK) monolayers displayed a 4-fold increase in accumulation in the basolateral to apical orientation relative to the apical to basolateral orientation, although no differential accumulation was noted between MDCK-WT and MDCK-MDR1 monolayers. Caco-2 monolayers demonstrated an approximate 2-fold increase in accumulation of MDMA. Exposure of various inhibitors of active drug transporters demonstrated mixed results; ritonavir, progesterone and indomethacin produced an approximately 50% reduction of MDMA transport, while verapamil, MK-571 and probenecid had no effect. Based on these data, it is concluded that MDMA efflux is mediated via the activity of a transporter distinct from P-glycoprotein. The possible inhibitory effects of amphetamines on rhodamine-123 transport were also assessed. MDMA, methylenedioxyamphetamine, amphetamine and methamphetamine, at physiologically relevant concentrations, did not significantly alter the transport of rhodamine-123 in Caco-2 monolayers or the LS180 cell line, suggesting that these compounds do not alter the function of P-glycoprotein.     

顺式-和反式-阿霍烯在Caco-2细胞模型中的体外摄取、转运和外排特性

研究顺式-阿霍烯(Z-Ajo)和反式-阿霍烯(E-Ajo)的肠细胞摄取、转运和外排特性。采用体外培养的人结肠Caco-2细胞单层模型评价，应用高效液相色谱法测定Z-Ajo和E-Ajo的含量。结果表明，仅能在Caco-2细胞单层的顶侧检测到Z-Ajo或E-Ajo；阿霍烯在Caco-2细胞中的代谢可被抗氧化剂维生素C、细胞色素P450药物代谢酶3A亚型抑制剂甲吡酮和ATP抑制剂叠氮化钠所抑制。Z-Ajo和E-Ajo皆不能透过Caco-2单层细胞而被迅速代谢，其代谢与CYP450药物代谢酶有关。     

Oleic acid decreases BCRP mediated efflux of mitoxantrone in Caco-2 cell monolayers

Breast cancer resistance protein (BCRP) efflux restricts intestinal absorption of substances like heterocyclic amines, mycotoxins and certain human and veterinary drugs. Fat rich meals seem to increase absorption of drugs which are BCRP substrates or inhibitors. We therefore hypothesize that absorption of toxicants normally effluxed by BCRP are increased by fatty acids in food. Transport across and accumulation of ³H-Mitoxantrone (MXR) in Caco-2 cell monolayers were measured after 60 min exposure to emulsions of ³H-MXR (1 μM) and oleic acid (0.5–5 mM). In addition, BCRP gene expression (RT-PCR) and the amount of BCRP protein (Western blot) were measured in oleic acid exposed Caco-2 cells. Oleic acid increased transport of MXR in a concentration dependent manner and 2 mM oleic acid or higher increased accumulation of MXR in cells, without any signs of cytotoxicity. Gene expression of BCRP was increased after exposure to oleic acid for 6 h, but the amount of BCRP protein was not increased. In conclusion, oleic acid clearly induced BCRP gene expression and reduced BCRP mediated efflux, although the amount of BCRP in cells was not affected. Consequently, effects of fatty acids on BCRP mediated efflux are important to consider in risk assessment of toxicants in food.     

Monoketocholate can decrease transcellular permeation of methotrexate across Caco‐2 cell monolayers and reduce its intestinal absorption in rat

Objectives Bile salts have been shown to decrease the absorption of methotrexate in the rat intestine by an unknown mechanism. We aimed to examine this effect. Methods We assessed apical‐to‐basolateral (AP‐BL) permeation of methotrexate (5 μM) across Caco‐2 cell monolayers pretreated with various concentrations (0, 0.25, 0.5, 1, 3 and 5 mm ) of sodium cholate or its semisynthetic analogue, sodium 12‐monoketocholate. We also determined the effect of orally administered 12‐monoketocholate on the intestinal absorption of methotrexate in rats to evaluate a possible in‐vitro–in‐vivo correlation. Key findings It was found that sodium cholate and sodium 12‐monoketocholate decreased the AP‐BL permeation of methotrexate at low concentrations (maximal inhibition at 0.25 and 1 mm , respectively) and increased it at higher concentrations. Determination of [¹⁴C] mannitol permeation and electrical resistance of monolayers during experiments showed that membrane integrity was not compromised at low concentrations of bile salts but was disrupted at higher concentrations. Subsequently, we examined the effect of the simultaneous oral administration of sodium 12‐monoketocholate (4, 20, 40 and 80 mg/kg) on the intestinal absorption of methotrexate in rats after an oral dose (5 mg/kg). The pharmacokinetic study showed that 12‐monoketocholate at 4 and 20 mg/kg did not change the methotrexate area under the serum concentration–time curve whereas sodium 12‐monoketocholate at 40 and 80 mg/kg significantly reduced it. Conclusions Sodium 12‐monoketocholate appears to decrease the intestinal absorption of methotrexate in rats by inhibition of transcellular active transport.     

蜕皮甾酮在Caco-2细胞模型中的摄取和跨膜转运研究

目的以Caco-2细胞单层模型,首次研究了蜕皮甾酮的口服吸收与转运特性。方法采用普通Caco-2细胞模型、表达活性CYP3A4酶的Caco-2细胞模型,分别从AP→BL方向和BL→AP方向研究蜕皮甾酮的摄取和跨膜转运规律。结果蜕皮甾酮在2种模型中的表观渗透系数(Papp)在0.1×10-6～1×10-6cm.s-1之间,药物吸收情况为1%～10%;在4 h的实验过程中,4种浓度蜕皮甾酮的ER值均小于1.5。结论研究表明,蜕皮甾酮主要以被动扩散的方式被细胞摄取和转运,其跨膜转运特性未受到CYP3A4-介导机制的影响。