title: Human immunodeficiency virus protease inhibitors interact with ATP binding cassette transporter 4/multidrug resistance protein 4: a basis for unanticipated enhanced cytotoxicity.
abstract: Human immunodeficiency virus (HIV) pharmacotherapy, by combining different drug classes such as nucleoside analogs and HIV protease inhibitors (PIs), has increased HIV-patient life expectancy. Consequently, among these patients, an increase in non-HIV-associated cancers has produced a patient cohort requiring both HIV and cancer chemotherapy. We hypothesized that multidrug resistance protein 4/ATP binding cassette transporter 4 (MRP4/ABCC4), a widely expressed transporter of nucleoside-based antiviral medications as well as cancer therapeutics might interact with PIs. Among the PIs evaluated (nelfinavir, ritonavir, amprenavir, saquinavir, and indinavir), only nelfinavir both effectively stimulated MRP4 ATPase activity and inhibited substrate-stimulated ATPase activity. Saos2 and human embryonic kidney 293 cells engineered to overexpress MRP4 were then used to assess transport and cytotoxicity. MRP4 expression reduced intracellular accumulation of nelfinavir and consequently conferred survival advantage to nelfinavir cytotoxicity. Nelfinavir blocked Mrp4-mediated export, which is consistent with its ability to increase the sensitivity of MRP4-expressing cells to methotrexate. In contrast, targeted inactivation of Abcc4/Mrp4 in mouse cells specifically enhanced nelfinavir and 9-(2-phosphonylmethoxyethyl) adenine cytotoxicity. These results suggest that nelfinavir is both an inhibitor and substrate of MRP4. Because nelfinavir is a new MRP4/ABCC4 substrate, we developed a MRP4/ABCC4 pharmacophore model, which showed that the nelfinavir binding site is shared with chemotherapeutic substrates such as adefovir and methotrexate. Our studies reveal, for the first time, that nelfinavir, a potent and cytotoxic PI, is both a substrate and inhibitor of MRP4. These findings suggest that HIV-infected cancer patients receiving nelfinavir might experience both enhanced antitumor efficacy and unexpected adverse toxicity given the role of MRP4/ABCC4 in exporting nucleoside-based antiretroviral medications and cancer chemotherapeutics.
authors.affiliation: Department of Pharmaceutical Sciences, St. Jude Children's Research Hospital, Memphis, Tennessee, USA.
authors.forename: Yu
authors.initials: Y
authors.lastname: Fukuda
authors.forename: Kazumasa
authors.initials: K
authors.lastname: Takenaka
authors.forename: Alex
authors.initials: A
authors.lastname: Sparreboom
authors.forename: Satish B
authors.initials: SB
authors.lastname: Cheepala
authors.forename: Chung-Pu
authors.initials: CP
authors.lastname: Wu
authors.forename: Sean
authors.initials: S
authors.lastname: Ekins
authors.forename: Suresh V
authors.initials: SV
authors.lastname: Ambudkar
authors.forename: John D
authors.initials: JD
authors.lastname: Schuetz
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology / Q000633:toxicity; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002460:Cell Line; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004347:Drug Interactions; D005260:Female; D017320:HIV Protease Inhibitors / Q000494:pharmacology* / Q000633:toxicity; D006801:Humans; D006868:Hydrolysis; D008727:Methotrexate / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008958:Models, Molecular; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D019888:Nelfinavir / Q000494:pharmacology / Q000633:toxicity; D063065:Organophosphonates / Q000494:pharmacology / Q000633:toxicity; D019438:Ritonavir / Q000494:pharmacology / Q000633:toxicity
chemical_list: D000225:Adenine; D000970:Antineoplastic Agents; D017320:HIV Protease Inhibitors; D008727:Methotrexate; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D019888:Nelfinavir; D063065:Organophosphonates; D019438:Ritonavir; C053001:adefovir