Characterization of midazolam metabolism in locusts: The role of CYP3A4-like enzyme in the formation of 1'-OH and 4-OH midazolam

TY - JOUR

T1 - Characterization of midazolam metabolism in locusts: The role of CYP3A4-like enzyme in the formation of 1'-OH and 4-OH midazolam

AU - Olsen, Line Rørbæk

AU - Gabel-Jensen, Charlotte

AU - Wubshet, Sileshi Gizachew

AU - Kongstad, Kenneth Thermann

AU - Janfelt, Christian

AU - Badolo, Lassina

AU - Hansen, Steen Honoré

PY - 2016/2/1

Y1 - 2016/2/1

N2 - 1. The metabolism of midazolam was investigated in vivo in locusts in order to evaluate the presence of an enzyme with functionality similar to human CYP3A4/5.2. Hydroxylated metabolites of midazolam identical to human metabolites were detected in locusts and the apparent affinities (Km values) were in the same range as reported in humans (in locusts: 7-23 and 33-85 M for the formation of the 1′-OH and 4-OH metabolites, respectively).3. The formation of hydroxylated metabolites could successfully be inhibited by co-administration of ketoconazole, a known CYP3A4/5 inhibitor.4. Besides phase I metabolites, a number of conjugated metabolites were detected using high-resolution mass spectrometry. The most abundant metabolites detected were structurally identified by 1H NMR as two N-glucosides. NMR analysis strongly suggested that the glycosylation occurred at the two nitrogens (either one in each case) of the imidazole ring.5. Distribution of midazolam and the glucose conjugates were successfully measured using desorption electrospray mass spectrometry imaging revealing time-dependent changes in distribution over time.6. In conclusion, it appears that an enzyme with functionality similar to human CYP3A4/5 is present in locusts. However, it appears that conjugation with glucose is the main detoxification pathway of midazolam in locusts.

AB - 1. The metabolism of midazolam was investigated in vivo in locusts in order to evaluate the presence of an enzyme with functionality similar to human CYP3A4/5.2. Hydroxylated metabolites of midazolam identical to human metabolites were detected in locusts and the apparent affinities (Km values) were in the same range as reported in humans (in locusts: 7-23 and 33-85 M for the formation of the 1′-OH and 4-OH metabolites, respectively).3. The formation of hydroxylated metabolites could successfully be inhibited by co-administration of ketoconazole, a known CYP3A4/5 inhibitor.4. Besides phase I metabolites, a number of conjugated metabolites were detected using high-resolution mass spectrometry. The most abundant metabolites detected were structurally identified by 1H NMR as two N-glucosides. NMR analysis strongly suggested that the glycosylation occurred at the two nitrogens (either one in each case) of the imidazole ring.5. Distribution of midazolam and the glucose conjugates were successfully measured using desorption electrospray mass spectrometry imaging revealing time-dependent changes in distribution over time.6. In conclusion, it appears that an enzyme with functionality similar to human CYP3A4/5 is present in locusts. However, it appears that conjugation with glucose is the main detoxification pathway of midazolam in locusts.

U2 - 10.3109/00498254.2015.1051604

DO - 10.3109/00498254.2015.1051604

M3 - Journal article

C2 - 26207435

SN - 0049-8254

VL - 46

SP - 99

EP - 107

JO - Xenobiotica

JF - Xenobiotica

IS - 2

ER -