title: In vitro and in vivo reversal of P-glycoprotein-mediated multidrug resistance by a novel potent modulator, XR9576.
abstract: The overexpression of P-glycoprotein (P-gp) on the surface of tumor cells causes multidrug resistance (MDR). This protein acts as an energy-dependent drug efflux pump reducing the intracellular concentration of structurally unrelated drugs. Modulators of P-gp function can restore the sensitivity of MDR cells to such drugs. XR9576 is a novel anthranilic acid derivative developed as a potent and specific inhibitor of P-gp, and in this study we evaluate the in vitro and in vivo modulatory activity of this compound. The in vitro activity of XR9576 was evaluated using a panel of human (H69/LX4, 2780AD) and murine (EMT6 AR1.0, MC26) MDR cell lines. XR9576 potentiated the cytotoxicity of several drugs including doxorubicin, paclitaxel, etoposide, and vincristine; complete reversal of resistance was achieved in the presence of 25-80 nM XR9576. Direct comparative studies with other modulators indicated that XR9576 was one of the most potent modulators described to date. Accumulation and efflux studies with the P-gp substrates, [3H]daunorubicin and rhodamine 123, demonstrated that XR9576 inhibited P-gp-mediated drug efflux. The inhibition of P-gp function was reversible, but the effects persisted for >22 h after removal of the modulator from the incubation medium. This is in contrast to P-gp substrates such as cyclosporin A and verapamil, which lose their activity within 60 min, suggesting that XR9576 is not transported by P-gp. Also, XR9576 was a potent inhibitor of photoaffinity labeling of P-gp by [3H]azidopine implying a direct interaction with the protein. In mice bearing the intrinsically resistant MC26 colon tumors, coadministration of XR9576 potentiated the antitumor activity of doxorubicin without a significant increase in toxicity; maximum potentiation was observed at 2.5-4.0 mg/kg dosed either i.v. or p.o. In addition, coadministration of XR9576 (6-12 mg/kg p.o.) fully restored the antitumor activity of paclitaxel, etoposide, and vincristine against two highly resistant MDR human tumor xenografts (2780AD, H69/LX4) in nude mice. Importantly all of the efficacious combination schedules appeared to be well tolerated. Furthermore, i.v. coadministration of XR9576 did not alter the plasma pharmacokinetics of paclitaxel. These results demonstrate that XR9576 is an extremely potent, selective, and effective modulator with a long duration of action. It exhibits potent i.v. and p.o. activity without apparently enhancing the plasma pharmacokinetics of paclitaxel or the toxicity of coadministered drugs. Hence, XR9576 holds great promise for the treatment of P-gp-mediated MDR cancers.
authors.affiliation: Xenova Limited, Slough, Berkshire, United Kingdom.
authors.forename: P
authors.initials: P
authors.lastname: Mistry
authors.forename: A J
authors.initials: AJ
authors.lastname: Stewart
authors.forename: W
authors.initials: W
authors.lastname: Dangerfield
authors.forename: S
authors.initials: S
authors.lastname: Okiji
authors.forename: C
authors.initials: C
authors.lastname: Liddle
authors.forename: D
authors.initials: D
authors.lastname: Bootle
authors.forename: J A
authors.initials: JA
authors.lastname: Plumb
authors.forename: D
authors.initials: D
authors.lastname: Templeton
authors.forename: P
authors.initials: P
authors.lastname: Charlton
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology*; D000166:Acridines / Q000494:pharmacology; D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology; D001386:Azides / Q000378:metabolism; D001667:Binding, Competitive / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D003524:Cyclosporins / Q000494:pharmacology; D003630:Daunorubicin / Q000378:metabolism / Q000494:pharmacology; D004095:Dihydropyridines / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000494:pharmacology; D018432:Drug Resistance, Multiple; D019008:Drug Resistance, Neoplasm; D004357:Drug Synergism; D005260:Female; D006801:Humans; D007546:Isoquinolines / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000473:pathology / Q000517:prevention & control; D017239:Paclitaxel / Q000493:pharmacokinetics; D011804:Quinolines / Q000494:pharmacology*; D044005:Tetrahydroisoquinolines; D013997:Time Factors; D016896:Treatment Outcome; D014316:Tritium; D023041:Xenograft Model Antitumor Assays
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000166:Acridines; D000903:Antibiotics, Antineoplastic; D000970:Antineoplastic Agents; D001386:Azides; D003524:Cyclosporins; D004095:Dihydropyridines; D007546:Isoquinolines; D011804:Quinolines; D044005:Tetrahydroisoquinolines; D014316:Tritium; C041130:azidopine; D004317:Doxorubicin; C402343:tariquidar; C083501:Elacridar; D017239:Paclitaxel; C070272:valspodar; D003630:Daunorubicin