Effect of black seed on dextromethorphan O- and N-demethylation in human liver microsomes and healthy human subjects.
OBJECTIVE
To investigate the effects of black seed on the metabolic activities of CYP3A4 and CYP2D6 in human liver microsomes and in human subjects using dextromethorphan as a probe drug.


METHODS
CYP2D6-mediated O-demethylation and CYP3A4-mediated N-demethylation of dextromethorphan (DEX) to dextrorphan (DOR) and 3-methoxymorphinan (3-MM), respectively, were utilized to assess the metabolic activities of the two enzymatic pathways. In the in vitro experiments, DEX was incubated with microsomes and NADPH in absence or presence of black seed extract (10-100 microg/ml) and the formation of the metabolites were measured by HPLC. In the clinical study, four healthy volunteers received a single oral dose of DEX 30 mg alone in phase I, and along with last dose of black seed (2.5 g twice daily for seven days) in phase II. Activities of the two enzymes were evaluated based on the urinary metabolic ratios (MRs), which were calculated from eight-hour urine collections. DEX and its metabolites were assayed in urine samples by HPLC following a liquid-liquid extraction.


RESULTS
Black seed extracts significantly inhibited the formation of both metabolites in microsomes. The maximum inhibition was observed at the highest extract concentration (i.e., 100 microg/ml), which was about 80% and 60% for DOR and 3-MM, respectively. In the clinical study, the urinary MRs of DEX/DOR and DEX/3-MM increased by factors of 127 and 1.6-fold, respectively, after consumption of black seed.


CONCLUSIONS
Black seed significantly inhibited CYP2D6 and CYP3A4 mediated metabolism of DEX in human liver microsomes and healthy human volunteers indicating that it has the potential to interact with CYP2D6 and CYP3A4 substrates.
Al-Jenoobi
F I
FI
Departments of Pharmaceutics, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia. aljenobi@ksu.edu.sa
Al-Thukair
A A
AA
Abbas
F A
FA
Ansari
M J
MJ
Alkharfy
K M
KM
Al-Mohizea
A M
AM
Al-Suwayeh
S A
SA
Jamil
S
S
D000293:Adolescent; D000328:Adult; D002851:Chromatography, High Pressure Liquid; D019389:Cytochrome P-450 CYP2D6 / Q000378:metabolism; D065690:Cytochrome P-450 CYP2D6 Inhibitors; D051544:Cytochrome P-450 CYP3A / Q000378:metabolism; D065692:Cytochrome P-450 CYP3A Inhibitors; D003915:Dextromethorphan / Q000031:analogs & derivatives / Q000378:metabolism / Q000652:urine; D003917:Dextrorphan / Q000378:metabolism / Q000652:urine; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology; D031881:Nigella sativa / Q000737:chemistry*; D010936:Plant Extracts / Q000008:administration & dosage / Q000494:pharmacology*; D012639:Seeds; D055815:Young Adult
D065690:Cytochrome P-450 CYP2D6 Inhibitors; D065692:Cytochrome P-450 CYP3A Inhibitors; D010936:Plant Extracts; D003917:Dextrorphan; C028657:3-methoxymorphinan; D003915:Dextromethorphan; D019389:Cytochrome P-450 CYP2D6; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human