Pharmacokinetics and biodistribution of the cyclin-dependent kinase inhibitor -CR8- in mice.
BACKGROUND
CR8 is a second generation inhibitor of cyclin-dependent kinases derived from roscovitine. CR8 was shown to be 50-100 fold more potent than roscovitine in inducing apoptosis in different tumor cell lines. In the present investigation, we have established an analytical method for the quantification of CR8 in biological samples and evaluated its bioavailability, biodistribution and pharmacokinetics in mice.


METHODS
A liquid chromatography method utilizing UV-detection was used for the determination of CR8. CR8 was administered either orally (100 mg/kg) or i.v. (50 mg/kg) and the animals were sacrificed at different time points. Blood samples and organs were collected, after which the pharmacokinetic parameters were calculated for plasma and organs.


RESULTS
CR8 was eluted at 5 minutes in the high performance liquid chromatography system used. The LLOQ detection was 0.10 μg/ml and linearity was observed within the 0.10-10 μg/ml range (r² > 0.998). The accuracy and precision were >86%, while the recovery from plasma was >95%. CR8 was stable for 2 months at room temperature in both solution and plasma. CR8 pharmacokinetics was fitted to a two-compartment open model after oral administration and to a one compartment model after i.v. injection. The elimination half-life was about 3 hours. Organ exposure to CR8 (expressed as % AUC organ vs. AUC plasma) was highest in liver (205%), adipose tissue (188%) and kidney (150%) and low in bone marrow (30%) and brain (15%) as compared to plasma. The oral bioavailability of CR8 was found to be essentially 100%.


CONCLUSIONS
We have developed a rapid and simple method for the analysis of CR8. CR8 pharmacokinetics pattern showed 100% bioavailability, long half-life and limited distribution to brain and bone marrow, which may allow systemic exposure higher than the IC₅₀ reported for cell death in tumor cell lines. CR8 displays favorable pharmacological properties and is therefore a good candidate for future clinical studies.
Experimental Cancer Medicine (ECM), Laboratory Medicine, Karolinska Institutet, 141 86, Stockholm, Sweden. moustapha.hassan@ki.se.
Hatem
H
Sallam
Ibrahim
I
El-Serafi
Laurent
L
Meijer
Moustapha
M
Hassan
D000284:Administration, Oral; D000818:Animals; D001682:Biological Availability; D002851:Chromatography, High Pressure Liquid; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors*; D004355:Drug Stability; D005260:Female; D007275:Injections, Intravenous; D057230:Limit of Detection; D051379:Mice; D008807:Mice, Inbred BALB C; D015394:Molecular Structure; D009928:Organ Specificity; D011687:Purines / Q000008:administration & dosage / Q000097:blood / Q000493:pharmacokinetics* / Q000494:pharmacology; D011725:Pyridines / Q000008:administration & dosage / Q000097:blood / Q000493:pharmacokinetics* / Q000494:pharmacology; D015203:Reproducibility of Results; D014018:Tissue Distribution
C532131:CR8 compound; D011687:Purines; D011725:Pyridines; D018844:Cyclin-Dependent Kinases