In vitro and in vivo evaluation of WK-X-34, a novel inhibitor of P-glycoprotein and BCRP, using radio imaging techniques.

Abstract

Overexpression of the multidrug resistance proteins P-glycoprotein (Pgp) and breast cancer resistance protein (BCRP) results in treatment failure of many malignancies including ovarian cancer. Dual inhibition of Pgp and BCRP may restore the sensitivity of resistant cells to anticancer drugs. We report the synthesis and characterization of a novel anthranilic-acid based Pgp and BCRP modulator, WK-X-34. In vitro inhibition of Pgp activity was evaluated using 99mTc-Sestamibi and daunorubicin accumulation in Pgp overexpressing human ovarian cancer cells (A2780/Adr) and its sensitive counterpart (A2780/wt). Interaction with BCRP was examined with a mitoxantrone-efflux assay in BCRP-overexpressing MCF7/mx cells, with flow cytometry. Interactions with the multidrug resistance associated proteins (MRP) were evaluated in transfected MRP1, MRP2 and MRP3 cell lines, using a 5-CFDA efflux assay. In vivo 99mTc-Sestamibi imaging of human ovarian cancer xenografts was used to evaluate the in vivo efficacy of WK-X-34 in mice. Daunorubicin accumulation in A2780/Adr cells was inhibited by WK-X-34 at nanomolar concentrations (IC50: 82.1 +/- 6 nM). WK-X-34 inhibited mitoxantrone accumulation in BCRP-overexpressing cells at micromolar concentrations (IC50 = 26.5 +/- 4.6 microM), whereas WK-X-34 did not significantly alter 5-CFDA accumulation in MRP transfected cells. In vivo, uptake of 99mTc-Sestamibi was significantly increased in A2780/Adr xenograft tumors, brain and intestine (AUCs(0-4h) 136%, 147% and 138%; p < 0.05) in mice dosed with WK-X-34 (20 mg/kg i.p.). WK-X-34 selectively modulates Pgp and BCRP in vitro and in vivo in multidrug resistant ovarian cancer cells, and thus may have potential utility in the treatment of multidrug resistant tumors.

• Authors: Veronika Jekerle, Werner Klinkhammer, Deborah A Scollard, Kerstin Breitbach, Raymond M Reilly, Micheline Piquette-Miller, Michael Wiese
• Journal: International journal of cancer (Int J Cancer) Vol. 119 Issue 2 Pg. 414-22 (Jul 15 2006) ISSN: 0020-7136 [Print] United States
• PMID: 16646006 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: 2006 Wiley-Liss, Inc.

Chemical References

• ABCG2 protein, human
• ATP Binding Cassette Transporter, Subfamily B, Member 1
• ATP Binding Cassette Transporter, Subfamily G, Member 2
• ATP-Binding Cassette Transporters
• Antineoplastic Agents
• Benzamides
• Multidrug Resistance-Associated Proteins
• Neoplasm Proteins
• Radiopharmaceuticals
• WK-X-34
• multidrug resistance-associated protein 3
• Technetium Tc 99m Sestamibi
• Mitoxantrone
• multidrug resistance-associated protein 1
• Daunorubicin

Topics

• ATP Binding Cassette Transporter, Subfamily B, Member 1 (antagonists & inhibitors, metabolism)
• ATP Binding Cassette Transporter, Subfamily G, Member 2
• ATP-Binding Cassette Transporters (antagonists & inhibitors, metabolism)
• Antineoplastic Agents (administration & dosage, metabolism, pharmacology)
• Area Under Curve
• Benzamides (administration & dosage, pharmacology)
• Cell Line, Tumor
• Daunorubicin (metabolism)
• Drug Resistance, Multiple
• Drug Resistance, Neoplasm
• Female
• Flow Cytometry
• Fluorescence
• Gene Expression Regulation, Neoplastic
• Humans
• Mitoxantrone (metabolism)
• Multidrug Resistance-Associated Proteins (drug effects, metabolism)
• Neoplasm Proteins (antagonists & inhibitors, metabolism)
• Ovarian Neoplasms (drug therapy, metabolism)
• Radiopharmaceuticals
• Technetium Tc 99m Sestamibi
• Time Factors
• Tissue Distribution
• Transplantation, Heterologous
• Up-Regulation