Inhibition of ABCG2/BCRP-mediated transport-correlation analysis of various expression systems and probe substrates

Zsolt Sáfár; Gábor Kecskeméti; Judit Molnár; Anita Kurunczi; Zoltán Szabó; Tamás Janáky; Emese Kis; Péter Krajcsi

doi:10.1016/j.ejps.2020.105593

Inhibition of ABCG2/BCRP-mediated transport-correlation analysis of various expression systems and probe substrates

Eur J Pharm Sci. 2021 Jan 1:156:105593. doi: 10.1016/j.ejps.2020.105593. Epub 2020 Oct 13.

Authors

Zsolt Sáfár¹, Gábor Kecskeméti², Judit Molnár³, Anita Kurunczi⁴, Zoltán Szabó⁵, Tamás Janáky⁶, Emese Kis⁷, Péter Krajcsi⁸

Affiliations

¹ Solvo Biotechnology, a Charles River Company, 52 Közép fasor, Szeged H-6726, Hungary. Electronic address: safar@solvo.com.
² Department of Medical Chemistry, Faculty of Medicine, University of Szeged, Dóm tér 8, Szeged H-6720, Hungary. Electronic address: kecskemeti.gabor@med.u-szeged.hu.
³ Solvo Biotechnology, a Charles River Company, 52 Közép fasor, Szeged H-6726, Hungary. Electronic address: molnar.judit@solvo.com.
⁴ Solvo Biotechnology, a Charles River Company, 52 Közép fasor, Szeged H-6726, Hungary. Electronic address: kurunczi@solvo.com.
⁵ Department of Medical Chemistry, Faculty of Medicine, University of Szeged, Dóm tér 8, Szeged H-6720, Hungary. Electronic address: szabo.zoltan@med.u-szeged.hu.
⁶ Department of Medical Chemistry, Faculty of Medicine, University of Szeged, Dóm tér 8, Szeged H-6720, Hungary. Electronic address: janaky.tamas@med.u-szeged.hu.
⁷ Solvo Biotechnology, a Charles River Company, 52 Közép fasor, Szeged H-6726, Hungary. Electronic address: kis@solvo.com.
⁸ Solvo Biotechnology, a Charles River Company, 52 Közép fasor, Szeged H-6726, Hungary; Solvo Biotechnology, a Charles River Company, 4-20 Irinyi J str, Budapest H-1117, Hungary; Faculty of Information Technology and Bionics, Pázmány Péter Catholic University, Práter str 50/a, Budapest H-1083, Hungary; Semmelweis University, Faculty of Health Sciences, Vas str 17, Budapest H-1088, Hungary. Electronic address: krajcsi@solvo.com.

PMID: 33059043
DOI: 10.1016/j.ejps.2020.105593

Abstract

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_m 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_max values was observed, with BCRP-HEK293 membranes possessing the largest dynamic range. IC₅₀ 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 IC₅₀ values in the BCRP-HEK293 membranes were within 3-fold in pairwise comparisons. Overall, BCRP-HEK293 membranes demonstrated the highest activity. The IC₅₀ values showed good concordance but substrate dependent inhibition was observed for some drugs.

Keywords: ABCG2; BCRP; HEK293-BCRP; substrate-dependent inhibition; vesicular transport.

MeSH terms

ATP Binding Cassette Transporter, Subfamily G, Member 2
ATP-Binding Cassette Transporters*
HEK293 Cells
Humans
Neoplasm Proteins* / genetics
Neoplasm Proteins* / metabolism
Topotecan

Substances

ABCG2 protein, human
ATP Binding Cassette Transporter, Subfamily G, Member 2
ATP-Binding Cassette Transporters
Neoplasm Proteins
Topotecan