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@ARTICLE{Schneider:859684,
author = {Schneider, Daniela and Bier, Dirk and Bauer, Andreas and
Neumaier, Bernd and Holschbach, Marcus},
title = {{I}nfluence of incubation conditions on microsomal
metabolism of xanthine-derived {A}1 adenosine receptor
ligands},
journal = {Journal of pharmacological $\&$ toxicological methods},
volume = {95},
issn = {1056-8719},
address = {New York, NY [u.a.]},
publisher = {Elsevier},
reportid = {FZJ-2019-00523},
pages = {16 - 26},
year = {2019},
abstract = {Introduction:In vitro metabolism models such as liver
microsomes represent an important tool for the development
of novel radioligands.Comparability and physiological
relevance of invitro metabolism data criticallydepend on the
careful evaluation and optimization of assay protocols. We
therefore investigated the influence ofincubation conditions
on the microsomal stability of xanthine-derived A1 adenosine
receptor (A 1AR) ligandswhich have been developed for
positron emission tomography (PET).Methods:Substrate
depletion assays using rat liver microsomes (RLM) were
performed for three analogouscompounds which differ with
regard to the metabolically vulnerable substituent at the
xanthine C8 position.Incubation
conditionswerevariedsystematically. Additionally, the
stability of the cofactor NADPH duringing cubation was
investigated.Results:Microsomal metabolism was strongly
influenced by buffer pH, organic solvents and preincubation
time.Substrate depletion values varied up to5-fold depending
on incubation matrix composition, however, the rankorder of
metabolic stability remained unchanged. Prolonged incubation
periods led to drastic loss in enzymeactivity which could
not be prevented by addition of metal chelators or
antioxidants. Cofactor NADPH
wasrapidlyoxidizedinmicrosomalmatrix,evenintheabsenceofcytochromeP450substrates.Discussion:Insummary,short
incubation times, precise pH control and minimal
concentrations of organic solvents are mandatory to obtain
reliable microsomal stability data. Furthermore, invitro
metabolic stability of thetested A 1 AR ligands varied
largely depending on the particular C8 substituent.
Consequently, structural modifications at the xanthine C8
position appear to be a promising strategy for the
improvement of A1AR PETradioligands},
cin = {INM-5 / INM-2},
ddc = {610},
cid = {I:(DE-Juel1)INM-5-20090406 / I:(DE-Juel1)INM-2-20090406},
pnm = {573 - Neuroimaging (POF3-573)},
pid = {G:(DE-HGF)POF3-573},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:30476620},
UT = {WOS:000454124000004},
doi = {10.1016/j.vascn.2018.11.005},
url = {https://juser.fz-juelich.de/record/859684},
}