Antibody binding shift assay for rapid screening of drug interactions with the human ABCG2 multidrug transporter

Telbisz, Ágnes; Hegedüs, Csilla; Özvegy‐Laczka, Csilla; Goda, Katalin; Várady, György; Takáts, Zoltán; Szabó, Eszter; Sorrentino, Brian P.; Váradi, András; Sarkadi, Balázs

doi:10.1016/j.ejps.2011.10.021

Cited by 36 publications

(34 citation statements)

References 40 publications

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“…29 Such a polyspecificity toward A-ring chalcones is consistent with that observed for different classes of flavonoidic and flavonoid-like inhibitors, assumed to bind to the same site or to overlapping sites, such as hydrophobic flavones,19–21 rotenoids,23 acridones,17 tariquidar derivatives,16 methoxy trans -stilbenes,22 and other chalcones 27–29. Polyspecificity of inhibitory sites is also consistent with the well known polyspecificity of ABCG252 and other multidrug ABC transporters toward large panels of transport substrates. It is worthwhile mentioning that the gain-of-function R482T/G mutation in ABCG2, which was found to extend the transport substrate panel to rhodamine 123 and anthracyclins,53 also allowed inhibitors such as GF120918 and imatinib to be transported 52…”

Section: Resultssupporting

confidence: 86%

“…Polyspecificity of inhibitory sites is also consistent with the well known polyspecificity of ABCG252 and other multidrug ABC transporters toward large panels of transport substrates. It is worthwhile mentioning that the gain-of-function R482T/G mutation in ABCG2, which was found to extend the transport substrate panel to rhodamine 123 and anthracyclins,53 also allowed inhibitors such as GF120918 and imatinib to be transported 52…”

Section: Resultssupporting

confidence: 79%

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New structure–activity relationships of chalcone inhibitors of breast cancer resistance protein: polyspecificity toward inhibition and critical substitutions against cytotoxicity

Rangel¹,

Winter²,

Gauthier³

et al. 2013

DDDT

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Adenosine triphosphate-binding cassette subfamily G member 2 (ABCG2) plays a major role in cancer cell multidrug resistance, which contributes to low efficacy of chemotherapy. Chalcones were recently found to be potent and specific inhibitors, but unfortunately display a significant cytotoxicity. A cellular screening against ABCG2-mediated mitoxantrone efflux was performed here by flow cytometry on 54 chalcone derivatives from three different series with a wide panel of substituents. The identified leads, with submicromolar IC 50 (half maximal inhibitory concentration) values, showed that the previously identified 2′-OH-4′,6′-dimethoxyphenyl, as A-ring, could be efficiently replaced by a 2′-naphthyl group, or a 3′,4′-methylenedioxyphenyl with lower affinity. Such a structural variability indicates polyspecificity of the multidrug transporter for inhibitors. At least two methoxyl groups were necessary on B-ring for optimal inhibition, but substitution at positions 3, 4, and 5 induced cytotoxicity. The presence of a large O-benzyl substituent at position 4 and a 2′-naphthyl as A-ring markedly decreased the cytotoxicity, giving a high therapeutic ratio, which constitutes a critical requirement for future in-vivo assays in animal models.

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Section: Resultssupporting

confidence: 86%

Section: Resultssupporting

confidence: 79%

New structure–activity relationships of chalcone inhibitors of breast cancer resistance protein: polyspecificity toward inhibition and critical substitutions against cytotoxicity

Rangel¹,

Winter²,

Gauthier³

et al. 2013

DDDT

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show abstract

“…Indeed, transported substrates such as prazosin, quercetin, or nilotinib (Telbisz et al, 2012) strongly stimulate the basal ATPase activity, then enhancing “coupled” ATPase activity. Our present results also show the lack of any effect by DNP-SG on the ABCG2 transporter, suggesting that glutathione conjugates are not transported by ABCG2.…”

Section: Discussionmentioning

confidence: 99%

ABCG2 is not able to catalyze glutathione efflux and does not contribute to GSH-dependent collateral sensitivity

Gauthier¹,

Özvegy‐Laczka²,

Sarkadi³

et al. 2013

Front. Pharmacol.

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ABCG2 is a key human ATP-binding cassette (ABC) transporter mediating cancer cell chemoresistance. In the case of ABCC1, another multidrug transporter, earlier findings documented that certain modulators greatly increase ABCC1-mediated glutathione (GSH) efflux and, upon depletion of intracellular GSH, induce “collateral sensitivity” leading to the apoptosis of multidrug resistant cells. Recently, it has been suggested that ABCG2 may mediate an active GSH transport. In order to explore if ABCG2-overexpressing cells may be similarly targeted, we first looked for the effects of ABCG2 expression on cellular GSH levels, and for an ABCG2-dependent GSH transport in HEK293 and MCF7 cells. We found that, while ABCG2 overexpression altered intracellular GSH levels in these transfected or drug-selected cells, ABCG2 inhibitors or transport modulators did not influence GSH efflux. We then performed direct measurements of drug-stimulated ATPase activity and 3H-GSH transport in inside-out membrane vesicles of human ABC transporter-overexpressing Sf9 insect cells. Our results indicate that ABCG2-ATPase is not modulated by GSH and, in contrast to ABCC1, ABCG2 does not catalyze any significant GSH transport. Our data suggest no direct interaction between the ABCG2 transporter and GSH, although a long-term modulation of cellular GSH by ABCG2 cannot be excluded.

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“…The assay is based on the phenomenon that the binding of a conformation-sensitive antibody UIC2/5D3 to ABCB1/ABCG2 (respectively) could be increased in the presence of an ABCB1/ABCG2 substrate or inhibitor interacting with the transporter [19,20]. Inhibitors of ABCB1/ ABCG2 are expected to cause higher UIC2/5D3 shift effects as compared to those caused by transported substrates [22]. In the ABCB1-and ABCG2-overexpressing cells (SW620 Ad300 and S1M1 80, respectively), volasertib was found to cause a UIC2/5D3 shift in a concentration dependent manner (Figure 3B and 3C), comparable to that mediated by known ABCB1 (crizotinib) and ABCG2 (axitinib) inhibitors.…”

Section: Discussionmentioning

confidence: 99%

Volasertib (BI 6727), a novel polo-like kinase inhibitor, reverses ABCB1 and ABCG2-mediated multidrug resistance in cancer cells

Poon²,

Chen³

et al. 2013

J Cancer Ther Res

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However, the most common mechanism of resistance is the active efflux of drugs by ATP-binding cassette (ABC) transporters including P-glycoprotein (ABCB1/P-gp), ABCC1/ MRP1 and ABCG2 [6]. These transporters play a key role in the energy-dependent cellular efflux of toxic agents. They are capable of recognizing and extruding a broad range of functionally and structurally unrelated compounds, thereby causing the MDR phenotype in various cancer types. An obvious strategy to restore drug sensitivity in MDR cancer cells caused by ABC drug transporters is to block transporter-mediated drug efflux. Over the past decade, tremendous efforts have been made to discover and synthesize such inhibitors/modulators. Numerous clinical trials have been performed to evaluate the combination of ABCB1/P-gp modulators with standard chemotherapy regimens in enhancing anticancer efficacy [7]. However, none of them has been successfully put into clinical use, partly because of their low potency and lack of specificity

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Antibody binding shift assay for rapid screening of drug interactions with the human ABCG2 multidrug transporter

Cited by 36 publications

References 40 publications

New structure–activity relationships of chalcone inhibitors of breast cancer resistance protein: polyspecificity toward inhibition and critical substitutions against cytotoxicity

New structure–activity relationships of chalcone inhibitors of breast cancer resistance protein: polyspecificity toward inhibition and critical substitutions against cytotoxicity

ABCG2 is not able to catalyze glutathione efflux and does not contribute to GSH-dependent collateral sensitivity

Volasertib (BI 6727), a novel polo-like kinase inhibitor, reverses ABCB1 and ABCG2-mediated multidrug resistance in cancer cells

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Antibody binding shift assay for rapid screening of drug interactions with the human ABCG2 multidrug transporter

Cited by 36 publications

References 40 publications

New structure&ndash;activity relationships of chalcone inhibitors of breast cancer resistance protein: polyspecificity toward inhibition and critical substitutions against cytotoxicity

New structure&ndash;activity relationships of chalcone inhibitors of breast cancer resistance protein: polyspecificity toward inhibition and critical substitutions against cytotoxicity

ABCG2 is not able to catalyze glutathione efflux and does not contribute to GSH-dependent collateral sensitivity

Volasertib (BI 6727), a novel polo-like kinase inhibitor, reverses ABCB1 and ABCG2-mediated multidrug resistance in cancer cells

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Product

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New structure–activity relationships of chalcone inhibitors of breast cancer resistance protein: polyspecificity toward inhibition and critical substitutions against cytotoxicity

New structure–activity relationships of chalcone inhibitors of breast cancer resistance protein: polyspecificity toward inhibition and critical substitutions against cytotoxicity