title: A phase I pharmacodynamic study of the effects of the cyclin-dependent kinase-inhibitor AZD5438 on cell cycle markers within the buccal mucosa, plucked scalp hairs and peripheral blood mononucleocytes of healthy male volunteers.
abstract: OBJECTIVE
  AZD5438 is a novel, orally bioavailable, cyclin-dependent kinase (CDK) inhibitor demonstrating preclinical pharmacodynamic (PD) effects on CDK substrates and active growth inhibition of human tumour xenografts. Clinical pharmacokinetic (PK) data shows its plasma t1/2 to be 1-3 h. The main purpose of the current study was to evaluate PD activity of single oral doses of AZD5438 in healthy volunteers. Twelve healthy male subjects received 10, 40 or 60 mg AZD5438 or placebo in a rotating placebo crossover study design. Rapidly proliferating normal tissues [buccal mucosa, peripheral blood mononucleocytes (PBMCs) and plucked scalp hair] were sampled pre-dosing, 1.5 h (tmax), +/-6 h post-dosing. The primary PD endpoint, phospho-retinoblastoma protein (pRb) levels in buccal biopsies (unit length labelling index) assessed by immunohistochemistry, was used as a biomarker of CDK activity.
  
  
  RESULTS
  Phospho-pRb levels were demonstrated to decrease in an epitope, dose- and time-dependent manner. Statistically significant reductions in the ratio phospho-pRb/total pRb were detected at 1.5 h post-dose compared to placebo for both 40 mg [S807-S811 epitope geometric least-squares mean (glsmean) ratio = 0.75, P = 0.014] and 60 mg AZD5438 (S807-S811 epitope glsmean ratio = 0.74, P = 0.011; T821 epitope glsmean ratio = 0.72, P = 0.031). No statistically significant differences were noted at 6 h post-dosing, indicating a close PK-PD relationship between AZD5438 and target inhibition. No effects attributable to AZD5438 were detectable on phospho-p27, p27, Ki67 in the buccal mucosa; or on phospho-pRb (S249-T252 epitope), phospho-p27 or Ki67 in the sheath cells of plucked scalp hair, raising issues about the appropriateness of different detection methods/tissues for use as PD biomarkers. In ex vivo stimulated PBMCs, statistically and near-statistically significant anti-proliferative effects, with the suggestion of a dose-response effect, were noted on the incorporation of [3H]-thymidine (stimulated/non-stimulated) at 10, 40 and 60 mg, compared to placebo, at 1.5 h post-dosing (glsmean ratio = 0.65, P = 0.019; 0.70, P = 0.056; 0.51, P = 0.001, respectively).
  
  
  CONCLUSIONS
  The modest PD effect, short plasma t1/2 and close PK-PD relationship suggest that multiple daily dosing or sustained release formulations at higher doses will be necessary for AZD5438 to achieve sustained inhibition of CDK in human cancers.
authors.affiliation: Edinburgh Cancer Centre, Western General Hospital, Crewe Road, Edinburgh, EH4 2XU, UK.
authors.forename: D Ross
authors.initials: DR
authors.lastname: Camidge
authors.forename: Mike
authors.initials: M
authors.lastname: Pemberton
authors.forename: Jim
authors.initials: J
authors.lastname: Growcott
authors.forename: Dereck
authors.initials: D
authors.lastname: Amakye
authors.forename: David
authors.initials: D
authors.lastname: Wilson
authors.forename: Helen
authors.initials: H
authors.lastname: Swaisland
authors.forename: Cheryl
authors.initials: C
authors.lastname: Forder
authors.forename: Robert
authors.initials: R
authors.lastname: Wilkinson
authors.forename: Kate
authors.initials: K
authors.lastname: Byth
authors.forename: Andrew
authors.initials: A
authors.lastname: Hughes
mesh_terms: D015415:Biomarkers / Q000032:analysis; D002453:Cell Cycle / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D018592:Cross-Over Studies; D050756:Cyclin-Dependent Kinase Inhibitor Proteins / Q000494:pharmacology*; D006197:Hair / Q000378:metabolism*; D006801:Humans; D007093:Imidazoles / Q000008:administration & dosage / Q000494:pharmacology*; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D008297:Male; D009061:Mouth Mucosa / Q000378:metabolism*; D010919:Placebos / Q000008:administration & dosage; D011743:Pyrimidines / Q000008:administration & dosage / Q000494:pharmacology*; D016160:Retinoblastoma Protein / Q000032:analysis; D012535:Scalp
chemical_list: D015415:Biomarkers; D050756:Cyclin-Dependent Kinase Inhibitor Proteins; D007093:Imidazoles; D010919:Placebos; D011743:Pyrimidines; D016160:Retinoblastoma Protein; C521840:AZD5438