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Impact of ABCB1 genetic polymorphisms on the interactions between novel dipyranone derivatives and P‐glycoprotein
Author(s) -
Wang YenHsiang,
Chen ChienYu,
Liu NaiYu,
Chang ChihShiang,
Hung ChinChuan
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.1064.4
Subject(s) - efflux , p glycoprotein , rhodamine 123 , calcein , wild type , chemistry , atp binding cassette transporter , microbiology and biotechnology , multiple drug resistance , biology , pharmacology , biochemistry , gene , mutant , transporter , membrane , antibiotics
Multidrug resistance (MDR) is one of the major obstacle in cancer treatment and over‐expression of P‐glycoprotein (P‐gp) is highly related to this issue. Genetic polymorphisms in P‐gp may influence clinical treatment outcomes. Therefore, this study aims to evaluate the effect of novel dipyranone derivatives on wild‐ and variant‐types human P‐gp efflux function and investigate underlying kinetic and molecular mechanisms. Recombinant P‐gp‐expressing HEK293 cell lines were established with the common polymorphisms in human ABCB1 gene. Real‐time RT‐PCR was performed to confirm the mRNA expression. Influence of dipyranone derivatives on P‐gp function was evaluated by calcein‐AM uptake assay, rhodamine 123 efflux assay and doxorubicin efflux assay. MDR1 shift assay and ATPase assay were conducted to evaluate the interaction between P‐gp and dipyranone derivatives. Among the tested compounds, compound A, B and C were the top three P‐gp inhibitors after primary screening. SRB assay showed that all of these three compounds had no cytotoxicity effect. However, wild‐type P‐gp mRNA expressions were reduced after treating compound B and C. Calcein‐AM uptake assay, rhodamine 123 efflux assay and doxorubicin efflux assay indicated that these three compounds significantly inhibited P‐gp efflux function. For rhodamine 123 efflux, compound A competitive inhibited wild‐type P‐gp, while compound B and C uncompetitive inhibited wild‐type P‐gp. In doxorubicin efflux, compound A and B modulated wild‐type P‐gp through uncompetitive inhibition, and compound C inhibited wild‐type P‐gp by competitive inhibition. Moreover, the common variants on ABCB1 gene would affect the P‐gp inhibition effect and interaction kinetics of compound B. Based on the results of MDR1 shift assay, these three compounds were not the substrates of P‐gp. However, compound A, B, and C influenced the P‐gp ATPase function and may interact with other P‐gp substrates as indicated by the ATPase assays. This study characterized the novel dipyranone derivatives, compound A, B and C as potential P‐gp inhibitors. The inhibition potency were affected by the common polymorphism haplotypes in ABCB1 gene, which implied that genotypes may take into consideration for further in vivo studies.