start-ver=1.4
cd-journal=joma
no-vol=8
cd-vols=
no-issue=
article-no=
start-page=41
end-page=44
dt-received=
dt-revised=
dt-accepted=
dt-pub-year=2019
dt-pub=20190731
dt-online=
en-article=
kn-article=
en-subject=
kn-subject=
en-title=
kn-title=A proposed simple screening method to determine relative contributions of CYP3A4 and CYP3A5 to drug metabolism in?vitro
en-subtitle=
kn-subtitle=
en-abstract=
kn-abstract=Purpose
The cytochrome P450 (CYP) 3A family of enzymes metabolize the majority of clinically used drugs. CYP3A4 and CYP3A5 are the two major CYP3A isoforms, but exhinbit different substrate specificity. The aim of this study was to establish a simple screening method to determine the relative contributions of CYP3A4 and CYP3A5 to drug metabolism in vitro.
Methods
A screening method was developed based on competitive inhibition using luciferin-PPXE (L-PPXE), a luminogenic CYP3A substrate. CYP3cide, tacrolimus, and midazolam were selected as standard compounds metabolized by CYP3A4 or CYP3A5. Nine clinically-used drugs were evaluated for their abilities to inhibit luminescence resulting from L-PPXE metabolism. Appropriate reaction conditions for the screening method were determined using recombinant CYP3A4 and CYP3A5.
Results
A significant decrease in luminescence resulting from L-PPXE metabolism by CYP3A4 and CYP3A5 was observed only for drugs reported to be metabolized by CYP3As. The substrate specificities of CYP3A4 or CYP3A5 for the proposed CYP3A substrates using our screening method were consistent with those of previous reports or available drug information from pharmaceutical companies. The reaction volume for this method was 50 ƒÊL, and the time required for the entire procedure was 70 min. Furthermore, this screening can be performed using a single tube with minimal training.
Conclusions
Through the establishment of our screening method in the present study, we are sure it is useful to determine the relative contributions of CYP3A4 and CYP3A5 to drug metabolism in vitro.
en-copyright=
kn-copyright=
en-aut-name=MatsumotoJun
en-aut-sei=Matsumoto
en-aut-mei=Jun
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=1
ORCID=
en-aut-name=NakamuraHiroyoshi
en-aut-sei=Nakamura
en-aut-mei=Hiroyoshi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=2
ORCID=
en-aut-name=SanSu Nwe
en-aut-sei=San
en-aut-mei=Su Nwe
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=3
ORCID=
en-aut-name=SatoHikari
en-aut-sei=Sato
en-aut-mei=Hikari
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=4
ORCID=
en-aut-name=TakezawaManami
en-aut-sei=Takezawa
en-aut-mei=Manami
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=5
ORCID=
en-aut-name=KishiRyuto
en-aut-sei=Kishi
en-aut-mei=Ryuto
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=6
ORCID=
en-aut-name=KitoYutaro
en-aut-sei=Kito
en-aut-mei=Yutaro
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=7
ORCID=
en-aut-name=SuganoJunko
en-aut-sei=Sugano
en-aut-mei=Junko
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=8
ORCID=
en-aut-name=IzukiMai
en-aut-sei=Izuki
en-aut-mei=Mai
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=9
ORCID=
en-aut-name=YanagisawaNao
en-aut-sei=Yanagisawa
en-aut-mei=Nao
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=10
ORCID=
en-aut-name=IkedaNaoki
en-aut-sei=Ikeda
en-aut-mei=Naoki
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=11
ORCID=
en-aut-name=SaitoYusuke
en-aut-sei=Saito
en-aut-mei=Yusuke
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=12
ORCID=
en-aut-name=KatoYoshinori
en-aut-sei=Kato
en-aut-mei=Yoshinori
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=13
ORCID=
en-aut-name=YamadaHarumi
en-aut-sei=Yamada
en-aut-mei=Harumi
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=14
ORCID=
en-aut-name=FujiyoshiMasachika
en-aut-sei=Fujiyoshi
en-aut-mei=Masachika
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=15
ORCID=
en-aut-name=AriyoshiNoritaka
en-aut-sei=Ariyoshi
en-aut-mei=Noritaka
kn-aut-name=
kn-aut-sei=
kn-aut-mei=
aut-affil-num=16
ORCID=
affil-num=1
en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
kn-affil=
affil-num=2
en-affil=Department of Pharmaceutical Sciences, International University of Health and Welfare
kn-affil=
affil-num=3
en-affil=Graduate School of Pharmaceutical Sciences, International University of Health and Welfare
kn-affil=
affil-num=4
en-affil=Department of Pharmaceutical Sciences, International University of Health and Welfare
kn-affil=
affil-num=5
en-affil=Department of Pharmaceutical Sciences, International University of Health and Welfare
kn-affil=
affil-num=6
en-affil=Department of Pharmaceutical Sciences, International University of Health and Welfare
kn-affil=
affil-num=7
en-affil=Department of Pharmaceutical Sciences, International University of Health and Welfare
kn-affil=
affil-num=8
en-affil=Department of Pharmaceutical Sciences, International University of Health and Welfare
kn-affil=
affil-num=9
en-affil=Department of Pharmaceutical Sciences, International University of Health and Welfare
kn-affil=
affil-num=10
en-affil=Department of Pharmaceutical Sciences, International University of Health and Welfare
kn-affil=
affil-num=11
en-affil=Department of Pharmaceutical Sciences, International University of Health and Welfare
kn-affil=
affil-num=12
en-affil=Department of Pharmaceutical Sciences, International University of Health and Welfare
kn-affil=
affil-num=13
en-affil=Department of Pharmaceutical Sciences, International University of Health and Welfare
kn-affil=
affil-num=14
en-affil=Department of Pharmaceutical Sciences, International University of Health and Welfare
kn-affil=
affil-num=15
en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
kn-affil=
affil-num=16
en-affil=Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
kn-affil=
en-keyword=CYP3A
kn-keyword=CYP3A
en-keyword=CYP3A5*3 allele
kn-keyword=CYP3A5*3 allele
en-keyword=P450 Glo Assay system
kn-keyword=P450 Glo Assay system
en-keyword=Pharmacokinetics
kn-keyword=Pharmacokinetics
en-keyword=Pharmacogenetics
kn-keyword=Pharmacogenetics
END