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@PHDTHESIS{Schneider:836190,
author = {Schneider, Daniela},
title = {{R}elevance of in vitro metabolism models for {PET}
radiotracer development},
school = {Universität zu Köln},
type = {Dr.},
reportid = {FZJ-2017-05313},
pages = {140},
year = {2017},
note = {Universität zu Köln, Diss., 2017},
abstract = {The application of positron emission tomography (PET) as a
tool for molecular imaging of the brain islimited by the
availability of suitable radiotracers. In radiotracer
development, success or failure of anovel compound is
determined by numerous physicochemical and pharmacological
factors of whichmetabolism represents a particularly
important one. Hence, an initial assessment of the
metabolicproperties of a candidate radiotracer at an early
stage of the development process prior to expensiveand
time-consuming animal studies is desirable. Various in vitro
metabolism models are available forthe evaluation of
candidate drugs in the pharmaceutical sector, however, since
drug and tracerapplications differ fundamentally (especially
with regard to the amount of administered substance)
thequestion rises whether these models are also capable of
providing relevant and valid data for theevaluation of
radiotracers. In the present work, this question was
examined using three xanthinederivedadenosine A1 receptor
(A1AR) ligands,
8-cyclopentyl-3-(3-fluoropropyl)-1-propylxanthine(CPFPX),
8-cyclobutyl-3-(3-fluoropropyl)-1-propylxanthine (CBX) and
3-(3-fluoropropyl)-8-(1-methylcyclobutyl)-1-propylxanthine
(MCBX) as model compounds. Radioligands for imaging
adenosinereceptors in vivo are of particular interest for
biomedical research since this receptor system has
beenassociated with numerous physiological functions (e.g.
regulation of sleep) and pathophysiologicalconditions (e.g.
Alzheimer’s and Parkinson’s disease). In vivo metabolic
stability and metabolitepatterns of the three A1AR ligands
were investigated in the rat model and compared to in vitro
datagenerated in rat liver microsomes. Prior to the
characterisation of the microsomal metabolism of thetest
compounds, the influence of incubation conditions on the in
vitro test results was assessed and themicrosomal assay was
optimised in order to provide physiologically relevant data.
Metabolic activity inliver microsomes was strongly
influenced by the pH value of the buffer medium and the type
andconcentration of the organic solvent used to solubilise
the test compounds. Variations of incubationmatrix
composition resulted in variations of individual substrate
depletion values of up to $500\%,whereas$ substrate
depletion ratios proved to be highly constant. In vitro
microsomal half-lives (t1/2) ofthe compounds were determined
and expressed as ratios for the purpose of later in vitro-in
vivocomparison. The t1/2 ratios (± SE) of CBX:CPFPX,
MCBX:CPFPX and CBX:MCBX were 3.1 ± 0.11,1.4 ± 0.029 and
2.2 ± 0.033. In vivo metabolic stability of the
18F-labelled compounds were assessed inanaesthetised rats
via blood analysis. Plasma clearance (CL) values were
calculated and expressed asinversed ratios to facilitate
direct comparison with in vitro t1/2 ratios. The inversed CL
ratios (± SE) of[18F]CBX:[18F]CPFPX, [18F]MCBX:[18F]CPFPX
and [18F]CBX:[18F]MCBX were 2.6 ± 0.12, 0.82 ± 0.019
and3.1 ± 0.15. In vitro t1/2 ratios deviated between 19 and
$71\%$ from inverse CL ratios. These deviationscan be
considered small in view of the reduced complexity of the
microsomal model and the multitudeof physiological
parameters affecting in vivo pharmacokinetics of a
substance. Visual comparison ofmetabolite profiles generated
in vitro and in vivo revealed a high degree of similarity.
In conclusion,both quantitative and qualitative aspects of
radiotracer metabolism could be reasonably well predictedby
microsomal data. This result encourages the implementation
of in vitro metabolism studies as anintegral part of PET
radiotracer development.},
cin = {INM-2},
cid = {I:(DE-Juel1)INM-2-20090406},
pnm = {571 - Connectivity and Activity (POF3-571)},
pid = {G:(DE-HGF)POF3-571},
typ = {PUB:(DE-HGF)11},
url = {https://juser.fz-juelich.de/record/836190},
}