Sáfár Zsolt, Kecskeméti Gábor, Molnár Judit, Kurunczi Anita, Szabó Zoltán, Janáky Tamás, Kis Emese, Krajcsi Péter
Solvo Biotechnology, a Charles River Company, 52 Közép fasor, Szeged H-6726, Hungary.
Department of Medical Chemistry, Faculty of Medicine, University of Szeged, Dóm tér 8, Szeged H-6720, Hungary.
Eur J Pharm Sci. 2021 Jan 1;156:105593. doi: 10.1016/j.ejps.2020.105593. Epub 2020 Oct 13.
BCRP / ABCG2 is a key determinant of pharmacokinetics of substrate drugs. Several BCRP substrates and inhibitors are of low passive permeability, and the vesicular transport assay works well in this permeability space. Membranes were prepared from BCRP-HEK293, MCF-7/MX, and baculovirus-infected Sf9 cells with (BCRP-Sf9-HAM), and without (BCRP-Sf9) cholesterol loading. K values for three substrates - estrone-3-sulfate, sulfasalazine, topotecan - correlated well between the four expression systems. In contrast, a 10-20-fold range in V values was observed, with BCRP-HEK293 membranes possessing the largest dynamic range. IC values of the different test systems were similar to each other, with 94.4% of pairwise comparisons being within 3-fold. Substrate dependent inhibition showed somewhat greater variation, as 81.4% of IC values in the BCRP-HEK293 membranes were within 3-fold in pairwise comparisons. Overall, BCRP-HEK293 membranes demonstrated the highest activity. The IC values showed good concordance but substrate dependent inhibition was observed for some drugs.
乳腺癌耐药蛋白/ABCG2是底物药物药代动力学的关键决定因素。几种乳腺癌耐药蛋白底物和抑制剂的被动通透性较低，而囊泡转运试验在这种通透性范围内效果良好。用（携带乳腺癌耐药蛋白的杆状病毒感染的Sf9细胞-HAM）和未（乳腺癌耐药蛋白-Sf9）加载胆固醇的方法，从表达乳腺癌耐药蛋白的人胚肾29细胞（BCRP-HEK293）、MCF-7/MX细胞以及杆状病毒感染的Sf9细胞制备膜。三种底物——硫酸雌酮、柳氮磺胺吡啶、拓扑替康——在这四种表达系统中的K值相关性良好。相比之下，观察到V值有10至20倍的范围，其中BCRP-HEK293膜具有最大的动态范围。不同测试系统的IC值彼此相似，94.4%的成对比较在3倍以内。底物依赖性抑制表现出稍大的变化，因为在成对比较中，BCRP-HEK293膜中81.4%的IC值在3倍以内。总体而言，BCRP-HEK293膜表现出最高活性。IC值显示出良好的一致性，但对某些药物观察到了底物依赖性抑制。
