Mycophenolic acid interaction with cyclosporine and tacrolimus in vitro and in vivo: evaluation of additive effects on rat blood lymphocyte function.

The effect of mycophenolic acid (MPA) in combination with either cyclosporine (CsA) or tacrolimus (TRL) on whole-blood lymphocyte function was assessed in vitro as well as in vivo. For the in vitro studies, rat whole blood was incubated with different concentrations of MPA together with CsA or TRL. In vivo, rats (n = 6 per group) were orally treated with 2.5 or 5 mg/kg of mycophenolate mofetil (MMF), either alone or in combination with 5 mg/kg CsA or 4 mg/kg TRL. Blood was obtained before and at different times after dosing. For both in vitro and in vivo studies, mitogen-stimulated whole blood was analyzed by flow cytometry to determine inhibition of expression of lymphocyte proliferation (proliferating cell nuclear antigen, PCNA) and T-cell activation (eg, CD25). Plasma MPA concentrations were measured by HPLC, and whole-blood CsA and TRL concentrations were quantified using LC-MS/MS. In vitro, low concentrations of 250 and 500 nM MPA acted additively with CsA and overadditively with TRL to suppress lymphocyte function, whereas higher MPA concentrations (1000 nM) in these combinations did not further increase inhibition compared with monotherapy with CsA or TRL alone. In vivo, the MPA AUC0-24 showed a dose-dependent increase. CsA and TRL AUC0-24 were not influenced by the MMF dose. Combination therapy increased inhibition of lymphocyte function compared with MMF monotherapy with a pronounced effect on PCNA compared with CD25. Significant differences between 2.5 and 5 mg/kg MMF in the combination groups were observed at 2 or 6 hours after dosing because of the maximal inhibitory effect on PCNA and CD25 expression (P < 0.05, ANOVA). However, in combination with TRL no different effects on the inhibition of CD25 expression were found between the 2 MMF doses. These novel data indicate that measurement of pharmacodynamic parameters of lymphocyte function in whole blood may help to monitor drug combination therapy and provide a rationale for drug reduction to minimize toxicity without compromising efficacy.