PP149— The Ceiba CoCkTail foR DRug hyDRoxylaTion PhenoTyPing in hisPaniC PoPulaTions F. De Andres; M.E.G. Naranjo; M. Sosa-Macias; B.P. Lazalde Ramos; E. Tarazona-Santos; A. LLerena; and CEIBA Consortium CICAB, Clinical Research Centre, Extremadura University Hospital, BADAJOZ, Spain; Centro Interdisciplinario de Investigacion para el Desarrollo Integral Regional del IPN Unidad Durango, CIIDIR-IPN, Durango; Laboratorio de Medicina Molecular, Unidad Academica de Medicina Humana y Ciencias de la Salud, Universidad Autonoma de Zacatecas, Zacatecas, Mexico; and Departamento de Biologia Geral, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil Introduction: Interethnic variability of drug metabolism has been demonstrated across Iberoamerican populations by the CEIBA Consortium. Drug-metabolizing enzymes genotype just predicts actual drug metabolic activity (phenotype) in the cases of poor metabolizers. For example, only 30% of ultrarapid metabolizers can be predicted from genotyping (Llerena et al., 2012). Moreover, according to EMA recommendations, development of phenotyping procedures for drug interactions studies and clinical research recommended (EMA, 2013). Therefore, a novel cocktail approach to measure metabolic activity (metabolic ratios) of the main CYP enzymes in just one experiment is developed and validated to be used in the study of Latin-American populations. Patients (or Materials) and Methods: Subjects were given low oral doses of 100-mg caffeine, 25-mg losartan, 20-mg omeprazole, and 30-mg dextromethorphan. Blood samples were taken 4 hours after administering the drugs to assay the following metabolic ratios in plasma: CYP1A2 (caffeine/paraxanthine), CYP2C9 (losartan /E-3174), CYP2C19 (omeprazole/5-hydroxyomeprazole), CYP2D6 (dextromethorphan/ dextrorphan), and CYP3A4 (dextromethorphan/3-methoxymorphinan). Solid phase extraction was utilized for analyte extraction and LC-MS/MS to quantify the probe drugs and metabolites. Results: Recovery values > 80% were obtained for all analytes, and no carryover or relative matrix effects were observed. The analytes were separated and detected in 9 minutes and the method was fully validated, with lower limits of quantification ranging from 0.3 ng mL–1 for 5-hydroxyomeprazole to 3.2 ng mL–1 for paraxanthine. The correlation coefficient (r2) values obtained were over 0.995 for all analytes. According to the EMA guideline on bioanalytical method validation, precision and accuracy values below 15% were achieved. Conclusion: The method was proven to be useful to measure targeted analytes for the evaluation of CYPs hydroxylation capacity in just a single experiment. Funding Sources: Supported by AEXCID-Cooperacion Extremena of Junta de Extremadura (11IA002) to Sociedad Ibero-Americana de Farmacogenetica-SIFF, and coordinated by the RIBEF network (Red Iberoamericana de Farmacogenetica y Farmacogenomica; www. ribef.com). Disclosure of Interest: None declared. PP151—PRevalenCe of CaRRiage CyP2C9, vkoRC1 anD CyP4f2 PolymoRPhisms in Russian PaTienTs WiTh high ThRomboTiC Risk PResCRibeD WaRfaRin ComPaReD WiTh oTheR eThniC gRouPs D. Ivashchenko; D. Sychev; I. Rusin; A. Grachev; and T. Beloshickaya Clinical Pharmacology, I.M. Sechenov First Moscow State Medical University; Cardiology; and Medical Genetic, SM-Clinic, Moscow, Russian Federation Introduction: VKORC1, CYP2C9, and CYP4F2 responsible for the metabolism of warfarin. Personal and ethnic differences in the genetic profile reveals in determining the outcome of a drug therapy. Aim: to explore the frequencies of CYP2C9, VCORC1, and CYP4F2 genotypes in Russians, compare results with ones for other nations Patients (or Materials) and Methods: A total 91 Caucasian subjects were recruited into the study. Forty (48.2%) patients were male and age was 66.17 (10.9) years. All patients had indications to receive warfarin. Medical records for the patients group were reviewed for the relevant clinical data. 5 ml of blood was taken from each subject, and DNA was isolated and used for identification of the CYP2C9 allele *1, *2, *3; G-1639A VCORC1; CYP4F2 V433M rs2108622 C > T using real-time polymerase chain reaction-restriction fragment length polymorphism assay. Results were compared with other ethnic groups and statistically analyzed with chi-square test. Results: We described the prevalence of CYP2C9 polymorphisms *1/*1 (67%), *1/*2 (9,9%), *1/*3 (11%), *2/*2 (2,2%), *2/*3 (8,8%), same for VKORC1 GG (49,5%), GA (28,6%), AA (22,%) and CYP4F2 CC (57,1%), CT (34,1%), TT (7,7%). No significant deviantions from Hardy-Wainberg equilibrium were observed (p 0.05). Conclusion: Prevalence of carriage of CYP2C9, VKORC1 polymorphisms in Russian patients with high thrombotic risk is closed to Caucasians and distinguish from Asians but no for CYP4F2. Disclosure of Interest: None declared.
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