title: Cassette dosing pharmacokinetics of a library of 2,6,9-trisubstituted purine cyclin-dependent kinase 2 inhibitors prepared by parallel synthesis.
abstract: Determination of pharmacokinetic properties in the intact animal remains a major bottleneck in drug discovery. Cassette dosing involves administration of a cocktail of drugs to individual animals. Here we describe the cassette dosing properties of a 107-membered library of 2,6,9-trisubstituted purine cyclin-dependent kinase 2 (CDK2) inhibitors. A three-step parallel synthesis approach produced compounds with purity ranging from 63% to 100%. Cassette dosing was validated by comparing the pharmacokinetic parameters obtained following i.v. administration of a mixture of olomoucine, R-roscovitine (CYC202), and bohemine, each at 16.6 mg/kg, with results for administration of single agents at 50 mg/kg. No significant difference was observed between the pharmacokinetic parameters of agents when dosed in combination compared with those of individual compounds. CYC202 showed the highest area under the curve (AUC) and the longest elimination half-life (t(1/2)). Further cassettes evaluated the library of trisubstituted purines with CYC202 and purvalanol A included as pharmacokinetic standards in a validated limited sampling strategy. The ratios of pharmacokinetic parameters to that of CYC202 [AUC, maximum concentration (C(max)), and t(1/2)] remained similar when compounds were tested in two different cassettes or as individual compounds. Following dosing of the same cassette on three different days, there was less than 20% variation in pharmacokinetic parameters between days. The structure-pharmacokinetics relationship showed that the favored purine substituents are benzylamine and veratrylamine at position 6, amino-2 propanol at position 2, and methylpropyl or hydroxyethyl at position 9. Without cassette dosing, this study would have used 3 times as many animals and would have taken 4 times longer, illustrating the power of this method in lead optimization.
authors.affiliation: Cancer Research UK Centre for Cancer Therapeutics, The Institute of Cancer Research, Sutton, Surrey, United Kingdom. Florence.Raynaud@icr.ac.uk
authors.forename: Florence I
authors.initials: FI
authors.lastname: Raynaud
authors.forename: Peter M
authors.initials: PM
authors.lastname: Fischer
authors.forename: Bernard P
authors.initials: BP
authors.lastname: Nutley
authors.forename: Phyllis M
authors.initials: PM
authors.lastname: Goddard
authors.forename: David P
authors.initials: DP
authors.lastname: Lane
authors.forename: Paul
authors.initials: P
authors.lastname: Workman
mesh_terms: D000818:Animals; D019540:Area Under Curve; D042846:CDC2-CDC28 Kinases / Q000037:antagonists & inhibitors*; D051357:Cyclin-Dependent Kinase 2; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000493:pharmacokinetics*; D005260:Female; D005609:Free Radicals; D015723:Gene Library; D051379:Mice; D008807:Mice, Inbred BALB C; D008956:Models, Chemical; D009369:Neoplasms / Q000188:drug therapy*; D011687:Purines / Q000737:chemistry* / Q000493:pharmacokinetics* / Q000494:pharmacology; D000077546:Roscovitine; D013329:Structure-Activity Relationship; D013997:Time Factors
chemical_list: C471843:6-((3-chloro)anilino)-2-(isopropyl-2-hydroxyethylamino)-9-isopropylpurine; D004791:Enzyme Inhibitors; D005609:Free Radicals; D011687:Purines; C118034:bohemine; D000077546:Roscovitine; D042846:CDC2-CDC28 Kinases; C495893:Cdk2 protein, mouse; D051357:Cyclin-Dependent Kinase 2