Clinical Therapeutics
Extremadura, Badajoz, Spain; 3Colegio de Ciencias de la Salud,
Universidad San Francisco de Quito, Quito, Ecuador; and
4
Servicio de Laboratorio, Hospital de los Valles, Quito, Ecuador
Introduction:  According to EMA recommendations, development
of phenotyping procedures for drug interactions studies and clinical
research are highly recommended,1 due to the discordances found
between genotypes determined of the main CYP enzymes and their
actual enzymatic activity.2 Recently, CEIBA cocktail approach to
measure metabolic activity of the main CYPs in just one experi-
ment has been designed.3 However, its optimal design remains to be
elucidated, which is the main objective of this research, in order to
appropriately select the sample to be analyzed and an optimal single
time point for the analysis, to avoid diminish costs and discomfort
to the volunteers without compromising the reliability of the results.
Material and Methods:  Thirteen healthy subjects were given low
oral doses of 100mg caffeine, 25mg losartan, 20mg omeprazole and
30 mg dextromethorphan, and blood and urine samples were taken
at different time points (predose, 0.5, 1, 2, 3, 4, 6, 8, and 12h) to
assay the concentration3 and pharmacokinetic parameters (AUC).
The MRs for CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4
were calculated at each time point and their correlation with AUC
ratios calculated. All subjects were genotyped.
Results:  The cocktail was well tolerated and single time point MRs
at 4h or 6h (only for CYP2C9) after dosing showed high correlations
with corresponding AUC ratios and can, therefore, be proposed as
simple phenotyping metrics. On the other hand, metabolic ratios in
urine samples were not comparable to plasma ratios.
Conclusion:  This cocktail was proven to reliably reflect the selected
CYPs activities for the evaluation of CYPs hydroxylation capacity.
The proposed simplified sampling scheme could facilitate clinical
application of CYPs phenotyping with one blood sample collection,
in a single analysis.
Supported by AEXCID of GOBEX (11IA002) to Sociedad
Iberoamericana de Farmacogenética, Instituto de Salud Carlos III-
Sara Borrell program (CD13/00348), and Collaboration grant from
USFQ (ET); coordinated by the RIBEF network.
Disclosure of Interest:  None Declared.
References
1. European Medicines Agency. Guideline on the investigation of drug
interactions. EMA/CHMP/EWP/125211/2010. 2012;1–60.
2. Llerena A, Dorado P, Ramírez R, et al. CYP2D6 genotype and
debrisoquine hydroxylation phenotype in Cubans and Nicaraguans.
Pharmacogenomics J. 2012;12:176–183.
3. De Andrés F, Sosa-Macías M, Llerena A. A rapid and simple LC–MS/
MS method for the simultaneous evaluation of CYP1A2, CYP2C9,
CYP2C19, CYP2D6 and CYP3A4 hydroxylation capacity. Bioanalysis.
2014;6:683–696.
Cyp450 Geno-Phenotype Analysis:
Application of Ceiba Cocktail To Mexican
Populations
F. de Andrés1; B.P. Lazalde-Ramos3; M. Sosa-Macías3; and
A. Llerena1,2
1
CICAB, Clinical Research Centre, Extremadura University
Hospital, Badajoz, Spain; 2Faculty of Medicine, University of
Extremadura, Badajoz, Spain; and 3Centro Interdisciplinario de
Investigación para el Desarrollo Integral Regional del IPN Unidad
Durango, Durango, México
Introduction:  For the CYP450 enzymes, there is significant variabil-
ity of actual activity within a genotype group, i.e. genotype does not
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always correlate with the functional phenotype.1 Therefore, the most
relevant genetic variants should be analyzed, together with the simul-
taneous analysis of the CYP450 isoforms hydroxylation capacities.
Considering this lack of accuracy and the void of pharmacogenetic
information in ethnically specific regions,2 this study aimed to explore
the relationship between the main CYP450 (i.e. CYP1A2, CYP2C9,
CYP2C19, and CYP2D6) polymorphisms and the metabolic ratios
(MRs) in a group of Mexican subjects.
Material and Methods:  Six hundred fifty-four healthy native individu-
als from Mexico were genotyped and additionally phenotyped by the
CEIBA cocktail.3 Plasma concentrations of the drugs and metabolites
were quantified and MRs calculated to determine the actual CYPs
metabolic capacity. The geno-phenotype relationship was evaluated by
correlation analysis between ‘activity score (AS)’ and logMR.
Results:  No adverse effects were reported. The prevalence of PM gen-
otypes is ≤ 2%, whereas for CYP2C19 and CYP2D6, an ultrarrapid
metabolizer prevalence of 3.7% and 12.8%, respectively, was found.
MRs correlated with AS for CYP2C9, CYP2C19 and CYP2D6, and
MRs varied across subjects with different AS. Those individuals with
2 or more active genes showed lower MRs than those with one or no
active genes. The frequency histograms and probit analysis showed
no clear bimodality. Not all the phenotypically PMs carried zero
active genes, whereas those individuals whose metabolism is faster
exhibit at least two active genes.
Conclusion:  This is the first study of simultaneous determination
of genotypes, phenotypes and its correlation analysis for different
CYPs on a Mexican population. The influence of the genetics on the
enzyme hydroxylation capacity is confirmed, though further research
on genotype-phenotype relationship is required due to the overlap
among different genotypes and that other factors can potentially
influence the activity of these enzymes.
Supported by AEXCID-GOBEX (11IA002) to SIFF, and ISCIII-
Sara Borrell program (CD13/00348). Coordinated by the RIBEF
network.
Disclosure of Interest:  None Declared.
References
1. Llerena A, Dorado P, Ramírez R, et al.CYP2D6 genotype and
debrisoquine hydroxylation phenotype in Cubans and Nicaraguans.
Pharmacogenomics J. 2012;12:176–183.
2. McGraw J, Waller D. Cytochrome P450 variations in different ethnic
populations. Expert Opin Drug Metab Toxicol. 2012;8:371–382.
3. De Andrés F, Sosa-Macías M, Llerena A. A rapid and simple LC–MS/
MS method for the simultaneous evaluation of CYP1A2, CYP2C9,
CYP2C19, CYP2D6 and CYP3A4 hydroxylation capacity. Bioanalysis.
2014;6:683–696.
Student Views on The Student-Run
Pharmacovigilance Program
T. Schutte1,2; J. Tichelaar1,2; M.O. Reumerman1; R. van Eekeren3,4;
S. Groenland1; L. Rolfes3,4; M.C. Richir1,2; E. van Puijenbroek3,4;
and M.A. van Agtmael1,2
1
VU University Medical Center, Amsterdam, the Netherlands;
2
RECIPE (Research & Expertise Center In Pharmacotherapy
Education), Amsterdam, the Netherlands; 3The Netherlands
Pharmacovigilance Centre Lareb (Lareb); and 4University of
Groningen, Groningen, The Netherlands
Background: Pharmacovigilance centers play a vital role in the
monitoring of drug safety after approval for marketing, and depend
mainly on the quantity and quality of reported adverse drug reac-
tions (ADRs). To ensure future ADR-reporting and increase
Volume 37 Number 8S

pharmacovigilance awareness among medical students, we developed
and aim to evaluate student outcomes of participation in our Student-
run Pharmacovigilance program.
Method:  A pilot study was performed in which student attitudes,
knowledge and skills on ADR-reporting were evaluated using an
e-questionnaire before and after participation in the student-run
pharmacovigilance program. Teams of (1st-4th year) medical stu-
dents assessed real ADR-reports from healthcare-professionals/
patients reported to Lareb. These student-assessments included a
causality assessment, (scientific) pharmacological explanation, feed-
back letter to the reporter, and summary for the pharmacovigilance-
databases of the European Medicines Agency and WHO.
Results:  From May 2014-January 2015, 100 different ADR-reports
selected by Lareb staff were handled by 43 students. Before participat-
ing in the pilot < 25% knew how to report ADRs. After participation
> 70% knew how to report, and were even able to name important
August 2015
Poster Presentations
details for ADR-reporting. Intention towards ADR-reporting was
assessed using a 7-point Likert scale (1: extremely unlikely-7:
extremely likely). Students indicated they intend to report serious
(6.44 SD 0.73) and unknown (6.44 SD 0.63) future encountered
ADRs. On a 5 point Likert scale students disagreed (2.50 SD 1.15)
their current curriculum covered pharmacovigilance well, found par-
ticipation educational (4.56 SD 0.63) and more interesting than fic-
tive casuistry (4.56 SD 0.73). Furthermore besides students reported
they learned how to asses ADRs, they stated to have learned skills/
knowledge regarding critical appraisal of information, understanding
of pharmacological mechanisms and scientific writing.
Conclusion:  The Student-run pharmacovigilance program is a valu-
able and novel educational experience. It creates awareness in future
doctors with the potential to increase ADR-reporting, lets students
practice in searching and writing scientifically and teaches the basics
of pharmacovigilance in real life.
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