Uptake of Clozapine into HL-60 Promyelocytic Leukaemia Cells

 

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The atypical antipsychotic, clozapine, exerts superior efficacy in therapy-resistant schizophrenia, but unfortunately induces agranulocytosis with an incidence of 0.8 - 1 %. In this study, we investigated the cellular uptake of clozapine into human promyelocytic leukaemia HL-60 cells using HPLC with electrochemical detection. On incubation with 1.25 to 40 µM clozapine for 30 min, a saturable, energy- and temperature-dependent uptake process takes place (K_m = 18.8 µM, k_cat = 1.36 nmol/5 min/mg protein at 37° C). This suggests membrane passage of clozapine by a carrier mechanism. 10 µM Indatraline, an inhibitor of dopamine, noradrenaline and serotonin (5-HT) reuptake, but not the selective 5-HT reuptake inhibitor fluvoxamine, markedly reduced the transport of clozapine by 62 %, whereas addition of 10 mM glucose to the incubation medium increased intracellular clozapine concentrations by 28 %. Since cyclosporine A, vinblastine or verapamil up to a final concentration of 10 µM did not alter the intracellular accumulation of clozapine, an involvement of P-glycoprotein seems to be unlikely. In summary, clozapine uptake into HL-60 cells meets criteria of an active unidirectional transport. Its molecular correlates remain to be established.

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Dr. Uwe Henning

Neurobiochemical Research Unit Department of Psychiatry
Heinrich-Heine-University Düsseldorf

Bergische Landstr. 2

40629 Düsseldorf

Germany

Email: KN27120@mail.lvr.de