Drug binding to P-glycoprotein is inhibited in normal tissues following SDZ-PSC 833 treatment.
Rats were treated with daily injections of SDZ-PSC 833 (PSC) to study the interaction of this potent modulator of multidrug resistance (MDR) with P-glycoprotein (P-gp) expressed in normal tissues. After 2 days of treatment, the level of P-gp expression, detected by Western blot analysis, was not modified in renal brush border membranes (BBMs) and brain capillaries. However, the amount of P-gp detected with the photoaffinity probe [125I]-arylazidoprazosin (IAAP) was decreased in both tissues, suggesting that the drug binding properties of P-gp were altered by PSC treatment. This effect was further characterized by treating rats with PSC for 10 days. Following these treatments, the amount of immunodetected P-gp was increased in renal BBMs and brain capillaries. However, no increase in P-gp expression was observed in photolabeling experiments, suggesting that induced P-gp was not functional. In vitro experiments performed with renal BBMs showed that the inhibition of P-gp photolabeling by cyclosporin A (CsA), verapamil and vinblastine could be reversed by performing washing steps to remove these drugs before incubating the samples with IAAP. However, the inhibition mediated by PSC was less reversible since photolabeling of P-gp remained inhibited following the washing steps. Pre-incubation of intact CHRC5 cells with PSC, CsA and verapamil also inhibited P-gp photolabeling and increased rhodamine 123 accumulation. For PSC pre-treated samples, these effects were not completely reversed following washing, but were abolished for CsA and Ver pre-treated samples. Our results suggest that PSC could block P-gp function by a different mechanism from that of CsA and verapamil, involving modification of the drug binding sites.
Jetté
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Département de Chimie-Biochimie, Université du Québec á Montréal, Hôpital Ste-Justine, Canada.
Murphy
G F
GF
Béliveau
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D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism*; D000284:Administration, Oral; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000378:metabolism / Q000494:pharmacology; D001386:Azides; D015153:Blotting, Western; D001921:Brain / Q000098:blood supply / Q000187:drug effects / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000378:metabolism / Q000494:pharmacology; D002196:Capillaries / Q000187:drug effects / Q000378:metabolism; D016572:Cyclosporine / Q000494:pharmacology; D003524:Cyclosporins / Q000494:pharmacology*; D004347:Drug Interactions; D005456:Fluorescent Dyes / Q000493:pharmacokinetics; D007279:Injections, Subcutaneous; D007457:Iodine Radioisotopes; D007668:Kidney / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008871:Microvilli / Q000378:metabolism; D019920:Photoaffinity Labels; D011224:Prazosin / Q000031:analogs & derivatives; D051381:Rats; D017207:Rats, Sprague-Dawley; D020112:Rhodamine 123; D012235:Rhodamines / Q000493:pharmacokinetics; D014700:Verapamil / Q000378:metabolism / Q000494:pharmacology; D014747:Vinblastine / Q000378:metabolism / Q000494:pharmacology
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000972:Antineoplastic Agents, Phytogenic; D001386:Azides; D002121:Calcium Channel Blockers; D003524:Cyclosporins; D005456:Fluorescent Dyes; D007457:Iodine Radioisotopes; D019920:Photoaffinity Labels; D012235:Rhodamines; D020112:Rhodamine 123; D014747:Vinblastine; D016572:Cyclosporine; C041552:azidoprazosin; D014700:Verapamil; C070272:valspodar; D011224:Prazosin