Leflunomide’s active metabolite teriflunomide inhibits dihydro-oroate dehydrogenase, an enzyme essential to proliferation of T lymphocytes. As teriflunomide must reach the target site to have this effect, this study assessed the distribution of teriflunomide into T lymphocytes, as intracellular concentrations may be a superior response biomarker to plasma concentrations. CD3 MicroBeads (Miltenyi Biotec, Bergisch Gladbach, Germany) were used to extract CD3+ T cells from the peripheral blood of patients with rheumatoid arthritis who were taking a stable dose of leflunomide. Unbound plasma and intra-CD3+ T cell teriflunomide concentrations were quantified using liquid chromatography–mass spectrometry. Concentration (log transformed) and partition differences were assessed through paired Student t tests. Sixteen patients provided plasma steady-state teriflunomide samples, and eight provided a sample 6–12 weeks later. At time-point one, the geometric mean teriflunomide concentration (range) in CD3+ T cells was 18.12 μg/L (6.15–42.26 μg/L) compared with 69.75 μg/L (32.89–263.1 μg/L) unbound in plasma (P < 0.001). The mean partition coefficient (range) for unbound plasma teriflunomide into CD3+ T cells was 0.295 (0.092–0.632), which was significantly different from unity (P < 0.001). The median (range) change in teriflunomide concentration between the two time points was 14% (−10% to 40%) in unbound plasma and −29% (−69 to 138%) for CD3+ T cells. Because teriflunomide concentrations in CD3+ T cells were lower and displayed a higher intraindividual variability than the unbound plasma concentrations, its applicability as a therapeutic drug-monitoring marker may be limited.
- Received June 6, 2016.
- Accepted October 13, 2016.
- Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics