Pharmacokinetics of anticancer drugs in vitro.
It is generally assumed that drug concentration does not change significantly under cell culture conditions. Nevertheless, most of the therapeutic trials in acute leukemia that were based on in vitro drug sensitivity assays of patient samples have been disappointing. In order to show possible pitfalls of unphysiological alterations in vitro we investigated concentration versus time curves, metabolism and effects on the culture media for some antineoplastic drugs. Oxazaphosphorines and cytarabine were incubated in RPMI and in established cell lines and measured by HPLC. HPLC also served to measure enzyme activity and levels of related amino acids at various concentrations of asparaginase, ammonia release was photometrically determined. Etoposide was monitored by HPLC relative to different contents of FCS in RPMI. All oxazaphosphorines showed a rapid decrease of in vitro activity down to about 10% within 4-6 h, and 2% within 72 h. The level of cytarabine, when incubated in RPMI, was stable over 24h, and no change was seen with K562, while a rapid decrease to below 50% occurred within 6h in the presence of HL 60 and BLIN. 2 U/L of asparaginase led to asparagine depletion of the medium within 4h, while 200 U/L were associated with a preferential increase of glutamic acid and ammonia. Further, there was evidence of instability by rapid adsorption to plastic surfaces (paclitaxel) or isomerisation (etoposide) in RPMI with low FCS content. The instability of drugs in vitro is attributed to a variety of different factors: i.e. physico-chemical instability results in inactivation of oxazaphosphorines, cytarabine disappears by cellular metabolism without saturation depending on the cell-line. Epiphenomena like adsorption and isomerisation in vitro are unphysiological. Results of drug sensitivity assays should be interpreted with great caution.
Wagner
A
A
Department of Pediatric Hematology and Oncology, WWU Münster, Germany. WAGNEAL@uni-muenster.de
Hempel
G
G
Gumbinger
H G
HG
Jürgens
H
H
Boos
J
J
D000596:Amino Acids / Q000032:analysis; D000970:Antineoplastic Agents / Q000493:pharmacokinetics*; D001215:Asparaginase / Q000493:pharmacokinetics; D003520:Cyclophosphamide / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D003561:Cytarabine / Q000493:pharmacokinetics; D005047:Etoposide / Q000493:pharmacokinetics; D018922:HL-60 Cells / Q000378:metabolism; D006801:Humans; D007938:Leukemia; D014407:Tumor Cells, Cultured / Q000378:metabolism*
D000596:Amino Acids; D000970:Antineoplastic Agents; D003561:Cytarabine; D005047:Etoposide; D003520:Cyclophosphamide; D001215:Asparaginase