Sudoh M, Pauletti G M, Yao W, Moser W, Yokoyama A, Pasternak A, Sprengeler P A, Smith A B, Hirschmann R, Borchardt R T
Department of Pharmaceutical Chemistry, The University of Kansas, Lawrence 66047, USA.
Pharm Res. 1998 May;15(5):719-25. doi: 10.1023/a:1011966918959.
To compare the permeation characteristics of amide bond-containing HIV-1 protease inhibitors and their pyrrolinone-containing counterparts across Caco-2 cell monolayers, a model of the intestinal mucosa.
Transepithelial transport and cellular uptake of three pairs of amide bond-containing and pyrrolinone-based peptidomimetics were assessed in the presence and absence of cyclosporin A using the Caco-2 cell culture model. The potential of the peptidomimetics to interact with biological membranes was estimated by IAM chromatography.
In the absence of cyclosporin A, apical (AP) to basolateral (BL) flux of all compounds studied was less than the flux determined in the opposite direction (i.e., BL-to-AP). The ratio of the apparent permeability coefficients (Papp) calculated for the BL-to-AP and AP-to-BL transport (P(BL-->AP)/P(AP-->BL)) varied between 1.7 and 36.2. When individual pairs were ompared, P(BL-->AP)/P(AP-BL) ratios of the pyrrolinone-containing compounds were 1.5 to 11.5 times greater than those determined for the amide bond-containing analogs. Addition of 25 microM cyclosporin A to the transport buffer reduced the P(BL-->AP)/P(AP-->BL) ratios for all protease inhibitors to a value close to unity. Under these conditions, the amide bond-containing peptidomimetics were at least 1.6 to 2.8 times more able to permeate Caco-2 cell monolayers than were the pyrrolinone-containing compounds. The intrinsic uptake characteristics into Caco-2 cells determined in the presence of 25 microM cyclosporin A were slightly greater for the amide bond-containing protease inhibitors than for the pyrrolinone-containing analogs. These uptake results are consistent with the transepithelial transport results determined across this in vitro model of the intestinal mucosa.
The amide bond-containing and pyrrolinone-based peptidomimetics are substrates for apically polarized efflux systems present in Caco-2 cell monolayers. The intrinsic permeabilities of the amide bond-containing protease inhibitors are slightly greater than the intrinsic permeabilities of the pyrrolinone-based analogs through Caco-2 cell monolayers.
比较含酰胺键的HIV-1蛋白酶抑制剂及其含吡咯烷酮类似物在Caco-2细胞单层（一种肠黏膜模型）中的渗透特性。
使用Caco-2细胞培养模型，在有和没有环孢素A存在的情况下，评估三对含酰胺键和基于吡咯烷酮的拟肽的跨上皮转运和细胞摄取。通过离子交换色谱法评估拟肽与生物膜相互作用的潜力。
在没有环孢素A的情况下，所有研究化合物的顶侧（AP）到基底外侧（BL）通量小于相反方向（即BL到AP）测定的通量。计算的BL到AP和AP到BL转运的表观渗透系数（Papp）之比（P（BL→AP）/P（AP→BL））在1.7至36.2之间变化。当比较各对时，含吡咯烷酮化合物的P（BL→AP）/P（AP - BL）比值比含酰胺键类似物测定的值大1.5至11.5倍。向转运缓冲液中添加25μM环孢素A将所有蛋白酶抑制剂的P（BL→AP）/P（AP→BL）比值降低至接近1的值。在这些条件下，含酰胺键的拟肽比含吡咯烷酮的化合物渗透Caco-2细胞单层的能力至少高1.6至2.8倍。在存在25μM环孢素A的情况下测定的Caco-2细胞内摄取特性，含酰胺键的蛋白酶抑制剂比含吡咯烷酮的类似物略高。这些摄取结果与在该肠黏膜体外模型中测定的跨上皮转运结果一致。
含酰胺键和基于吡咯烷酮的拟肽是Caco-2细胞单层中存在的顶侧极化外排系统的底物。含酰胺键的蛋白酶抑制剂通过Caco-2细胞单层的固有渗透率略高于基于吡咯烷酮的类似物的固有渗透率。
