Binding of the epoxide cryptophycin analog, LY355703 to albumin and its effect on in vitro antiproliferative activity.
Cryptophycin, isolated from the cyanobacterium Nostoc, is a cytotoxic dioxadiazacyclohexadecenetetrone which causes rapid depletion of microtubules in intact cells. In the present report, the effect of protein binding of a new synthetic cryptophycin analog, LY355703 (cryptophycin 52), is discussed. In handling the compound, it was found to bind extensively to surfaces, and a high degree of plasma protein binding was also observed (about 99% in human plasma). Similarly, while LY355703 displays potent antiproliferative activity against several human tumor cell lines in vitro (IC50s ranging from 12 to 40 pM), the addition of human or bovine serum albumin (BSA) to CCRF-CEM cells adapted to serum-free (UltraCHO) medium markedly reduced its anti-proliferative activity. For example, the IC50s for LY355703 in BSA at 0, 4 and 40 mg/ml were 2, 19 and 34 pM, respectively. In comparison, the IC50 only increased 2-fold (4210-8530 pM) for taxol over the same BSA concentration range. When log phase CCRF-CEM cells were exposed to 1 microM [3H]LY355703, there was a rapid accumulation of drug, so that LY355703 reached steady state within 10 min. The rate of LY355703 uptake in log-phase CCRF-CEM human leukemia cells was a linear function of concentration over a wide range (0.25-50 microM), although the cytotoxicity IC50 was 19 pM. Drug accumulation was not inhibited by sodium azide. Although cryptophycin was observed to bind extensively to albumin, binding did not markedly modulate cryptophycin uptake by CCRF-CEM cells. Overall, these results demonstrate that attention must be given to the binding properties of LY355703 and similar cryptophycins while handling these compounds, and that binding to albumin (and probably other cellular components as well) is a significant factor for interpretation of results both in vitro and in vivo.
Cancer Research Division, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
R M
RM
Schultz
C
C
Shih
P G
PG
Wood
S D
SD
Harrison
W J
WJ
Ehlhardt
D000818:Animals; D000970:Antineoplastic Agents / Q000493:pharmacokinetics* / Q000633:toxicity; D001692:Biological Transport; D001943:Breast Neoplasms; D002417:Cattle; D002470:Cell Survival / Q000187:drug effects*; D047630:Depsipeptides; D004285:Dogs; D005260:Female; D006801:Humans; D007700:Kinetics; D007769:Lactams / Q000493:pharmacokinetics* / Q000633:toxicity*; D007783:Lactones / Q000493:pharmacokinetics* / Q000633:toxicity*; D007938:Leukemia; D008175:Lung Neoplasms; D051379:Mice; D017239:Paclitaxel / Q000633:toxicity; D011485:Protein Binding; D051381:Rats; D012709:Serum Albumin / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000378:metabolism; D014316:Tritium; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D047630:Depsipeptides; D007769:Lactams; D007783:Lactones; D012709:Serum Albumin; D014316:Tritium; D012710:Serum Albumin, Bovine; C114033:cryptophycin 52; D017239:Paclitaxel