Elucidation of N 1-methyladenosine as a Potential Surrogate Biomarker for Drug Interaction Studies Involving Renal Organic Cation Transporters

Abstract

Endogenous substrates are emerging biomarkers for drug transporters, which serve as surrogate probes in drug-drug interaction (DDI) studies. In this study, the results of metabolome analysis using wild-type and Oct1/2 double knockout mice suggested that N ¹-methyladenosine (m¹A) was a novel organic cation transporter (OCT) 2 substrate. An in vitro transport study revealed that m¹A is a substrate of mouse Oct1, Oct2, Mate1, human OCT1, OCT2, and multidrug and toxin exclusion protein (MATE) 2-K, but not human MATE1. Urinary excretion accounted for 77% of the systemic elimination of m¹A in mice. The renal clearance (46.9 ± 4.9 ml/min per kilogram) of exogenously given m¹A was decreased to near the glomerular filtration rates by Oct1/2 double knockout or Mate1 inhibition by pyrimethamine (16.6 ± 2.6 and 24.3 ± 0.6 ml/min per kilogram, respectively), accompanied by significantly higher plasma concentrations. In vivo inhibition of OCT2/MATE2-K by a single dose of 7-[(3R)-3-(1-aminocyclopropyl)pyrrolidin-1-yl]-1-[(1R,2S)-2-fluorocyclopropyl]-8-methoxy-4-oxoquinoline-3-carboxylic acid in cynomolgus monkeys resulted in the elevation of the area under the curve of m¹A (1.72-fold) as well as metformin (2.18-fold). The plasma m¹A concentration profile showed low diurnal and interindividual variation in healthy volunteers. The renal clearance of m¹A in younger (21-45 year old) and older (65-79 year old) volunteers (244 ± 58 and 169 ± 22 ml/min per kilogram, respectively) was about 2-fold higher than the creatinine clearance. The renal clearances of m¹A and creatinine were 31% and 17% smaller in older than in younger volunteers. Thus, m¹A could be a surrogate probe for the evaluation of DDIs involving OCT2/MATE2-K. SIGNIFICANCE STATEMENT: Endogenous substrates can serve as surrogate probes for clinical drug-drug interaction studies involving drug transporters or enzymes. In this study, m¹A was found to be a novel substrate of renal cationic drug transporters OCT2 and MATE2-K. N ¹-methyladenosine was revealed to have some advantages compared to other OCT2/MATE substrates (creatinine and N ¹-methylnicotinamide). The genetic or chemical impairment of OCT2 or MATE2-K caused a significant increase in the plasma m¹A concentration in mice and cynomolgus monkeys due to the high contribution of tubular secretion to the net elimination of m¹A. The plasma m¹A concentration profile showed low diurnal and interindividual variation in healthy volunteers. Thus, m¹A could be a better biomarker of variations in OCT2/MATE2-K activity caused by inhibitory drugs.