Pharmacological interaction with sunitinib is abolished by a germ‐line mutation (1291T>C) of BCRP/ABCG2 gene

Abstract

Sunitinib malate (Sutent, SU11248) is a small-molecule multitargeted tyrosine kinase inhibitor (TKI) used for the treatment of renal cell carcinoma and imatinib-resistant gastrointestinal stromal tumors. Some TKIs can overcome multidrug resistance conferred by ATP-binding cassette transporter, P-glycoprotein (P-gp) ⁄ ABCB1, multidrug resistance-associated protein 1 (MRP1) ⁄ ABCC1, and breast cancer resistance protein (BCRP) ⁄ ABCG2. Here, we analyzed the effects of sunitinib on P-gp and on wild-type and germ-line mutant BCRPs. Sunitinib remarkably reversed BCRP-mediated and partially reversed P-gp-mediated drug resistance in the respective transfectants. The in vitro vesicle transport assay indicated that sunitinib competitively inhibited BCRP-mediated estrone 3-sulfate transport and P-gp-mediated vincristine transport. These inhibitory effects of sunitinib were further analyzed in Q141K-, R482G-, R482S-, and F431L-variant BCRPs. Intriguingly, the F431L-variant BCRP, which is expressed by a germ-line mutant allele 1291T>C, was almost insensitive to both sunitinib-and fumitremorgin C (FTC)-mediated inhibition in a cell proliferation assay. Sunitinib and FTC did not inhibit 125 I-iodoarylazidoprazosin-binding to F431L-BCRP. Thus, residue Phe-431 of BCRP is important for the pharmacological interaction with sunitinib and FTC. Collectively, this is the first report showing a differential effect of a germ-line variation of the BCRP ⁄ ABCG2 gene on the pharmacological interaction between small-molecule TKIs and BCRP. These findings would be useful for improving our understanding of the pharmaceutical effects of sunitinib in personalized chemotherapy. (Cancer Sci 2010; 101: 1493-1500 T he ATP-binding cassette (ABC) transporter proteins, particularly P-glycoprotein (P-gp ⁄ ABCB1), multidrug resistance-associated protein 1 (MRP1 ⁄ ABCC1), and breast cancer resistance protein (BCRP ⁄ MXR ⁄ ABCP ⁄ ABCG2) have been extensively studied as key molecules that are involved in the multidrug-resistant phenotype of cancer cells.(1,2) P-gp effluxes various anticancer agents including vincristine (VCR), paclitaxel (PTX), doxorubicin (DOX), and mitoxantrone (MXR). (1,3) BCRP is referred to as a half-type ABC transporter that functions as a homodimer and transports anticancer agents such as topotecan, irinotecan, SN-38 (7-ethyl-10-hydroxycamptothecin), methotrexate, and MXR out of cells. Many compounds have been tested for their ability to overcome ABC transporter-mediated drug resistance. Verapamil, cyclosporine A (CsA), and other compounds have been identified as inhibitors of P-gp, (5,6) while fumitremorgin C (FTC), tamoxifen derivatives, and certain flavonoids inhibit BCRP.(7-10) Verapamil, for example, directly interacts with P-gp and competitively interferes with transporter-substrate binding. (11) Co-administration of inhibitory compounds would be expected to overcome unwanted anticancer drug resistance during chemotherapy, but is also suspected to affect the pharmacokinetics and pharmacodynamics of substrate anticancer drugs....

The Need for Human Breast Cancer Resistance Protein Substrate and Inhibition Evaluation in Drug Discovery and Development: Why, When, and How?

“…FTC, a mycotoxin, is one of the most potent BCRP inhibitors. Its inhibition potential on E3S and MTX in BCRP vesicles has been previously reported (0.28 mM and 0.30 mM, respectively; Chen et al, 2003;Kawahara et al, 2010) and is identical to our measured value. Our IC 50 values for imatinib, topotecan, rapamycin, eltrombopag, pantoprazole, and omeprazole are also very comparable to published results (Breedveld et al, 2004;Houghton et al, 2004;Ozvegy-Laczka et al, 2004;Gupta et al, 2006;Allred et al, 2011).…”

Interaction of innovative small molecule drugs used for cancer therapy with drug transporters

“…In contrast to the study by Thomas et al (2009), in this study the ABCG2 c.421C>A polymorphism was not associated with erlotinib disposition (Rudin et al, 2008). HEK293 cells (Shukla et al, 2009), K562 cells (Kawahara et al, 2010), LLC-PK1 cells (Hu et al, 2009) MRP2 (ABCC2) Docetaxel Saos2 cells (Hu et al, 2009) MRP4 (ABCC4) PMEA Saos2 cells (Hu et al, 2009) BCRP (ABCG2) Estrone-3-sulfate (Ki = 0.32 mM), Hoechst 33342, methotrexate, pheophorbide A, rhodamine 123 HEK293 cells (Dai et al, 2009;Shukla et al, 2009), K562 cells (Kawahara et al, 2010), Saos2 cells (Hu et al, 2009), S1-M1-80 cells (Dai et al, 2009) (Li et al, 2007) c.421C>A Increased drug accumulation (ratio of trough concentrations at steady state to day 1 trough concentration; malignant solid tumours) (Li et al, 2007) c.421C>A Increased risk of diarrhea (non-small-cell lung cancer) (Cusatis et al, 2006) c.421C>A No influence on risk of skin toxicity (non-small-cell lung cancer) (Cusatis et al, 2006) c.421C>A No effect on adverse side effects for example, diarrhea, interstitial pneumonia (non-small-cell lung cancer) (Akasaka et al, 2010) -15622TT Increased risk of diarrhea (non-small-cell lung cancer) (Lemos et al, 2011) haplotype TT (c.1143C>T, -15622C>T)…”

“…Sunitinib inhibits P-glycoprotein and BCRP function in vitro with possible consequences for bioavailability of coadministered drugs and for reversing efflux transportermediated multidrug resistance in humans (Dai et al, 2009;Hu et al, 2009;Shukla et al, 2009;Kawahara et al, 2010). Interestingly, a germ-line mutation in ABCG2 (c.1291T>C) is almost insensitive to sunitinib-mediated inhibition in a cell proliferation assay (Kawahara et al, 2010).…”

Genetic Polymorphisms Associated with a Prolonged Progression-Free Survival in Patients with Metastatic Renal Cell Cancer Treated with Sunitinib

“…Furthermore, sunitinib reversed ABCG2-mediated multidrug resistance by inhibiting the drug efflux function of ABCG2 (30). This inhibitory capacity of sunitinib on ABCG2 appeared to be sensitive for the ABCG2 1291T > C genotype (31). In addition, another SNP in ABCG2 (421C > A) was associated with increased sunitinib exposure (32).…”