Mouse pharmacokinetics and metabolism of the curcumin analog, 4-piperidinone,3,5-bis[(2-fluorophenyl)methylene]-acetate(3E,5E) (EF-24; NSC 716993).
OBJECTIVE
Curcumin, a keto-enol constituent of turmeric, has in vitro and in vivo antitumor activity. However, in vivo potency is low due to poor oral absorption. The mono-carbonyl analog, 3,5-bis[(2-fluorophenyl)methylene]-4-piperidinone acetate (EF-24, NSC 716993), exhibited broad-spectrum activity in the NCI anticancer cell line screen and potent antiangiogenesis activity in a HUVEC cell migration assay. The purpose of this study was to characterize the preclinical pharmacology of EF-24 in mice.


METHODS
EF-24 plasma stability, protein binding, pharmacokinetics, and metabolism were characterized utilizing an LC/MS/MS assay.


RESULTS
An LC/MS/MS assay incorporated protein precipitation with methanol, reverse-phase HPLC separation under gradient elution using an aqueous methanol mobile phase containing 0.1 % formic acid, and positive electrospray ionization detection of the m/z 312 > 149 transition for EF-24. The assay was linear over the range 7.8-1,000 nM. Plasma protein binding was >98 % with preferential binding to albumin. EF-24 plasma disposition in mice after i.v. administration of a 10 mg/kg dose was best fit to a 3-compartment open model. The terminal elimination half-life and plasma clearance values were 73.6 min and 0.482 L/min/kg, respectively. EF-24 bioavailability was 60 and 35 % after oral and i.p. administration, respectively. NADPH-dependent metabolism of EF-24 loss in liver microsomal preparations yielded several metabolites consistent with EF-24 hydroxylation and reduction.
Department of Oncology, Division of Oncology Research, Mayo Clinic, 200 First Street SW, Rochester, MN, 55905, USA, reid@mayo.edu.
Joel M
JM
Reid
Sarah A
SA
Buhrow
Judith A
JA
Gilbert
Lee
L
Jia
Mamoru
M
Shoji
James P
JP
Snyder
Matthew M
MM
Ames
D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D003474:Curcumin / Q000031:analogs & derivatives* / Q000378:metabolism / Q000493:pharmacokinetics*; D004285:Dogs; D006801:Humans; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D010881:Piperidones / Q000378:metabolism / Q000493:pharmacokinetics; D011485:Protein Binding; D051381:Rats
D010881:Piperidones; D003474:Curcumin