Abstract
Background
Human mass balance studies and the assessment of absolute oral bioavailability (F) are usually assessed in separate studies. Intravenous microdose administration of an isotope tracer concomitant to an unlabeled oral dose is an emerging technique to assess F. We report a novel double-tracer approach implemented for tofogliflozin combining oral administration of a radiolabel tracer with concomitant intravenous administration of a stable isotope tracer. Tofogliflozin is a potent and selective sodium/glucose cotransporter 2 inhibitor for the treatment of type 2 diabetes mellitus currently in clinical development.
Objectives
The objectives of the present study were to assess the systemic exposure of major circulating metabolites, excretion balance, F and contribution of renal clearance (CLR) to total clearance (CL) of tofogliflozin in healthy subjects within one study applying a novel double-tracer technique.
Methods
Six healthy male subjects received 20 mg [12C/14C]tofogliflozin (3.73 MBq) orally and a concomitant microdose of 0.1 mg [13C]tofogliflozin intravenously. Pharmacokinetics of tofogliflozin were determined for the oral and intravenous route; the pharmacokinetics of the metabolites M1 and M5 were determined for the oral route. Quantification of [12C]tofogliflozin in plasma and urine and [13C]tofogliflozin in plasma was performed by selective LC-MS/MS methods. For the pre-selected metabolites of tofogliflozin, M1 and M5, a validated liquid chromatography–tandem mass spectrometry (LC-MS/MS) was applied to plasma and urine samples. Total radioactivity was assessed in plasma, urine and feces. Pharmacokinetic analysis was conducted by non-compartmental methods.
Results
The pharmacokinetics of tofogliflozin in healthy subjects were characterized by an F of 97.5 ± 12.3 %, CL of 10.0 ± 1.3 l/h and volume of distribution at steady-state (V ss) of 50.6 ± 6.7 l. The main route of elimination of total drug-related material was by excretion into urine (77.0 ± 4.1 % of the dose). The observed CLR of 25.7 ± 5.0 ml/min was higher than the product of the estimated glomerular filtration rate (eGFR) and fraction unbound in plasma (f u) (eGFR × f u 15 ml/min), indicating the presence of net active tubular secretion in the renal elimination of tofogliflozin. However, CLR contributed only 15.5 % to the CL of tofogliflozin, suggesting that reductions in CLR by renal impairment won’t significantly affect systemic exposure to tofogliflozin. Tofogliflozin and its metabolite M1 were the only major circulating entities accounting for 46 ± 8.6 and 50 ± 8.2 %, respectively, of total circulating drug-related material, while the metabolite M5 was a minor circulating metabolite accounting for 3.0 ± 0.3 % of total circulating drug-related material. Both the M1 and M5 metabolites were excreted into urine and the major metabolite M1 did not exhibit active tubular secretion.
Conclusions
These results demonstrate the utility of the double-tracer approach to provide essential pharmacokinetic data and excretion data for drug-related material in one study at the same dosing occasion. The data obtained allowed the characterization of absorption, distribution, metabolism and excretion of tofogliflozin. Tofogliflozin exhibited highly favorable pharmacokinetic properties as demonstrated by its high F, low CL and a low V ss. The presence of only one major circulating metabolite of tofogliflozin was unambiguously demonstrated. As a drug targeting the kidney, luminal exposure of the kidney is achieved by renal filtration and active tubular secretion.
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Acknowledgments
Dietmar Schwab designed the study, approved the protocol, supervised the analysis and drafted the manuscript. Agnes Portron contributed to the protocol, analysis and interpretation of data. Zoe Backholer contributed to the protocol and acquisition of data. Berthold Lausecker (present address: CRS Clinical Research Services Mannheim GmbH, Germany) contributed to the design of the stable isotopes and supervised the LC-MS analysis. Kosuke Kawashima provided structural identification data of metabolites required for the pre-specified quantitative measurements in this study. All authors read and approved the final manuscript.
The authors want to acknowledge the many people involved in the conduct of the clinical trial, in particular the responsible investigator involved in the study, Dr. Stuart Mair (Quotient Clinical, Edinburgh, EH14 4AP, UK). Furthermore, the authors want to acknowledge Guy Fischer and Dr. Andreas Gloge (F. Hoffmann-LaRoche AG, pRED Non-clinical safety, Basel, Switzerland) for their bioanalytical support, Dr. Thomas Hartung (F. Hoffmann-LaRoche AG, Process Research & Synthesis, Basel, Switzerland) for the synthesis of the tofogliflozin isotopes, and Mizuki Yamane, Keiichi Morita, Tsutomu Sato and Yoshihito Ohtake (Chugai Pharmaceutical Co. Ltd., Research Division, Gotemba, Japan) for metabolite identification and reference material supply. The authors want to acknowledge the valuable discussions and input from Bruno Reigner (F. Hoffmann-LaRoche AG, pRED Clinical Pharmacology, Basel, Switzerland).
This study was funded by F. Hoffmann-La Roche AG. The authors are or were employed by F. Hoffmann-La Roche AG (Dietmar Schwab, Agnes Portron, Berthold Lausecker, Zoe Backholer) or by Chugai Pharmaceutical Co., Ltd. (Kosuke Kawashima).
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Schwab, D., Portron, A., Backholer, Z. et al. A Novel Double-Tracer Technique to Characterize Absorption, Distribution, Metabolism and Excretion (ADME) of [14C]Tofogliflozin After Oral Administration and Concomitant Intravenous Microdose Administration of [13C]Tofogliflozin in Humans. Clin Pharmacokinet 52, 463–473 (2013). https://doi.org/10.1007/s40262-013-0051-z
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DOI: https://doi.org/10.1007/s40262-013-0051-z