Influence of Tyrosine Kinase Inhibition on Organic Anion Transporting Polypeptide 1B3-Mediated Uptake

Abstract

The organic anion transporting polypeptide family member (OATP) 1B3 is a hepatic uptake transporter that has a broad substrate recognition and plays a significant role in regulating elimination of endogenous biomolecules or xenobiotics. OATP1B3 works in tandem with OATP1B1, with which it shares approximately 80% sequence homology and a high degree of substrate overlap. Despite some substrates being recognized solely by OATP1B3, its ability to compensate for loss of OATP1B1-mediated elimination and recognition by regulatory agencies, little is known about OATP1B3 regulatory factors and how they are involved with drugdrug interaction. It was recently discovered that OATP1B1 function is mediated by the activity of a particular tyrosine kinase that is sensitive to a variety of tyrosine kinase inhibitors (TKIs). This study reports that OATP1B3 is similarly regulated, as at least 50% of its activity is reduced by 20 US Food and Drug Administration -approved TKIs. Nilotinib was assessed as the most potent OATP1B3 inhibitor among the investigated TKIs, which can occur at clinically relevant concentrations and acted predominantly through noncompetitive inhibition without impacting membrane expression. Finally, OATP1B3 function was determined to be sensitive to the knockdown of the Lck/Yes novel tyrosine kinase that is sensitive to nilotinib and has been previously implicated in mediating OATP1B1 activity. Collectively, our findings identify tyrosine kinase activity as a major regulator of OATP1B3 function which is sensitive to kinase inhibition. Given that OATP1B1 is similarly regulated, simultaneous disruption of these transporters can have drastic effects on systemic drug concentrations, which would promote adverse events. SIGNIFICANCE STATEMENTThe organic anion transporting polypeptide family member (OATP) 1B3 is a facilitator of hepatic drug elimination, although much is unknown of how OATP1B3 activity is mediated, or how such regulators contribute to drug-drug interactions. This study reports that OATP1B3 activity is dependent on the Lck/Yes novel tyrosine kinase, which is sensitive to numerous tyrosine kinase inhibitors. These findings provide insight into the occurrence of many clinical drug-drug interactions, and a rationale for future study of tyrosine kinases regulating drug disposition.

Assessing Trans-Inhibition of OATP1B1 and OATP1B3 by Calcineurin and/or PPIase Inhibitors and Global Identification of OATP1B1/3-Associated Proteins

“…Among these, 861 and 357 proteins were specifically associated with FLAG-OATP1B1 and FLAG-OATP1B3, respectively, while they were not detected in Mock cells ( Figure 5 B). Trans -inhibition of OATP1B1 and/or OATP1B3 has been reported either previously or herein for the tyrosine kinase inhibitors (TKIs) nilotinib [ 13 , 14 ] and dasatinib [ 11 ], the mTOR kinase inhibitors sirolimus and everolimus [ 10 , 15 ], and the calcineurin and/or PPIase inhibitors CsA, tacrolimus, and SCY-635. Table 2 summarizes identified OATP1B1- and OATP1B3-accociated proteins related to biological processes and proteins relevant to these reported preincubation-induced inhibition effects.…”

“…OATP1B1 and OATP1B3 are phosphorylated proteins [ 7 , 12 ]. Some kinase modulator drugs/compounds such as protein kinase C activator [ 7 ] and tyrosine kinase inhibitor nilotinib [ 13 , 14 ] down-regulate OATP1B1/3 transport function under trans -inhibition conditions via modulating kinase activity. Many of the inhibitors with trans -inhibitory effects on OATP1B1/3 were also reported to cause time-dependent inhibition of OATP1B1/3 [ 3 , 4 , 8 , 9 , 10 , 11 , 15 , 16 ], where inhibitor preincubation enhances the inhibitory effect of the inhibitor, resulting in a reduced IC 50 /inhibition constant (K i ).…”

“…To date, global identification of OATP1B1- and OATP1B3-associated proteins through mass spectrometry-based proteomics has not been reported. As many drugs with reported trans -inhibitory effects on OATP1B1/3 are kinase inhibitors [ 10 , 11 , 13 , 14 , 65 ], the identification of proteins associated with OATP1B1 and OATP1B3 will not only broaden our understanding to transporter function and regulation in general, it may also shed light on potential mechanism(s) underlying the trans -inhibition of OATP1B1/3.…”

Itraconazole-Induced Increases in Gilteritinib Exposure Are Mediated by CYP3A and OATP1B

“…Surprisingly, these data suggest that gilteritinib noncompetitively inhibits OATP1B transport of E β G. Nonetheless, this observation is consistent with other OATP1B substrates [ 18 ], and is potentially due to the presence of multiple binding sites on OATP1B1 [ 19 ]. The potent inhibition of OATP1B1 by gilteritinib may be related to inhibition of the protein kinase LYN [ 20 , 21 , 22 ], which mediates phosphorylation-mediated activation of OATP1B1 and OATP1B3. Washout experiments demonstrated that the inhibitory mechanism observed for gilteritinib was reversible, with 75% of transport recovered within a 1-h period ( Figure 2 D).…”

Impact of Drug-Mediated Inhibition of Intestinal Transporters on Nutrient and Endogenous Substrate Disposition…an Afterthought?

“…These drugs can potentially disrupt BSEP-mediated bile acid secretion and promote cholestasis through the hepatic accumulation of bile acids [ 8 ]. Some TKIs have recently been shown to inhibit the activity of a variety of other transporters in a non-competitive manner [ 9 , 10 ]. Moreover, many drugs are associated with adverse events that can be partially explained by nutrient deficiencies.…”