W Arasihathesiscovershe Et

Total Page:16

File Type:pdf, Size:1020Kb

W Arasihathesiscovershe Et OpenAIR@RGU The Open Access Institutional Repository at Robert Gordon University http://openair.rgu.ac.uk Citation Details Citation for the version of the work held in ‘OpenAIR@RGU’: WARASIHA, B., 2008. Cytochrome P450 mRNA profile in human breast cancer cell lines. Available from OpenAIR@RGU. [online]. Available from: http://openair.rgu.ac.uk Copyright Items in ‘OpenAIR@RGU’, Robert Gordon University Open Access Institutional Repository, are protected by copyright and intellectual property law. If you believe that any material held in ‘OpenAIR@RGU’ infringes copyright, please contact [email protected] with details. The item will be removed from the repository while the claim is investigated. CYTOCHROME P450 mRNA PROFILE IN HUMAN BREAST CANCER CELL LINES BENJAMART WARASIHA A thesis submitted in partial fulfilment of the requirements of The Robert Gordon University for the degree of Doctor of Philosophy School of Pharmacy and Life Sciences, The Robert Gordon University, Aberdeen. United Kingdom. May 2008 Abstract Abstract Cytochrome P450 enzymes (P450s) are involved in cancer development and treatment due to their roles in the oxidative metabolism of various endogenous (e.g. oestrogen) and exogenous (e.g. tamoxifen) compounds. It is well-known that intermediate P450 metabolites derived from oestrogen metabolism are associated with breast carcinogenesis. The main aim of this project was to profile the cytochrome P450 and P450-regulatory nuclear receptor mRNAs in a series of breast cancer cell lines (BCCs) and compare this profile with normal breast cells. This study used the qualitative reverse transcriptase- polymerase chain reaction (RT-PCR) to detect mRNA expression of target genes. Results showed CYP1B1, CYP2D6, CYP2J2, CYP2R1, CYP2U1 and CYP4X1 mRNA to be present in all cell lines. CYP2A6, CYP2C8, CYP2C18, CYP2F1 and CYP4Z1 mRNA were expressed in oestrogen receptor (ER)-positive Caucasian and ER-negative Afro- Caribbean BCCs. Although no differences in P450 mRNA were observed between the different ethnic groups, these preliminary findings suggest potential similarities in the ER- positive Caucasian and ER-negative Afro-Caribbean BCCs which warrant further investigation The CYP4Z1 PCR product was identified as two distinct bands. Specific primer sets were used to demonstrate potential intron retention in CYP4Z1. Using established in vitro models for the study of regulatory mechanisms of CYP4Z1, T47D and ZR-75-1 breast cancer cell lines were used to determine the appropriate nuclear receptors (i.e. progesterone receptor, glucocorticoid receptor or peroxisome proliferator-activated receptor alpha ). These findings suggest that there may be an alternative receptor mechanism involved in CYP4Z1 mRNA induction in these cells. In conjunction, pre-treatment of these two cell lines with the RNA synthesis inhibitor actinomycin D followed by the agonists showed a significant reduction ( p < 0.05) of CYP4Z1 mRNA levels and inhibited CYP4Z1 induction by either progesterone, dexamethasone or pirinixic acid, indicating that these agonists have effects on CYP4Z1 mRNA transcription or stability. In contrast, cycloheximide differentially affected the level of CYP4Z1 mRNA induction by these agonists. Taken together, these results suggest that CYP4Z1 mRNA induction in T47D and ZR-75-1 is mediated through differential cell type specific regulatory mechanisms and there is evidence for differential regulation of the splice variants. ii Abstract Keywords: Cytochrome P450 mRNA, cytochrome P450-regulatory nuclear receptor mRNA, Oestrogen receptor (ER)-positive and negative breast cancer cell lines (BCCs), Caucasian and Afro-Caribbean BCCs, CYP4Z1 alternative splicing, regulation of CYP4Z1 iii Acknowledgements Acknowledgements I wish to express my sincere gratitude to the following people for their help, support, guidance and encouragement during the course of my studies. My Director of Studies, Dr Morag McFadyen, for introducing me to the world of cytochrome P450s, her valuable suggestions, patience and encouragement throughout my PhD studies. Dr Rachel Knott, for her constant guidance, support and participating as my supervisor. Professor Angela Drake-Holland, for her helpful suggestions and discussion on the project. Mrs Dorothy Moir, for helping me out with practical support in the molecular biology laboratory, her wonderful friendship and motivation. Many thanks to Professor Mark Noble, Dr Graeme Kay and Dr Kennedy Nelson for taking the time to critically evaluate my thesis. Professor Terry Healey, Head of the School of Pharmacy and Life Sciences, for giving me the opportunity to pursue my studies. Dr Susanne Boyle, for her support and advice in the first year of my studies. Mr Brian De Jonckheere, for information technology assistance. All of the technicians in the School of Pharmacy who have helped with my project and been wonderful friends. Mrs Elizabeth Tomchak, my English tutor, for helping me improve my English skills. Mr Martin Simpson, the Research Degrees officer, for his support and assistance. Associate Professor Veerapol Kukongwiriyapan from the Department of Pharmacology, Faculty of Medicine, Khon Kaen University, Thailand, for his support and guidance. Sarah Garry, Michelle Frazier, Eimear Molloy and Ian Lonergan, Honours students at the School of Pharmacy and Life Sciences, for the results of the preliminary work in Chapter 4. Dr Colin Thompson, Mr Ray Mulrooney, Dr Bridgeen McCaughan, Mrs Julie Young, Miss Anita Weidman, Miss Nicola Silva, Mr Vibhu Paudyal and all other colleagues for sharing their time and friendship both in and out of the laboratory. To Anusorn Thiangsungnoen, Pattarin Kittiboonyakun, Sansanee Wangvoralak, Sirichai Mongkolkiatsri and all of the other Thai students in Aberdeen for their wonderful friendship and encouragement and for making me feel less homesick. To everyone in the Taekwondo and Badminton clubs at The Robert Gordon University, for their friendship. Special thanks to my Taekwondo instructor Miss Lyndzie Jefferey, whose classes always helped me to unwind and relax, no matter how stressful my day in the lab had been. iv Acknowledgements To my wonderful parents, Samruay and Sudjai Warasiha who have taught me never to give up, for their constant care, unconditional love, continuous support and encouragement throughout my life. My two brothers, Taweesit and Supakit Warasiha, and two sisters Somjit Nampha and Jurairat Sriwicha including their families for their understanding, encouragement, support and love. My boyfriend, Tim Cushnie, for his help and encouragement and for sharing my frustration and happy moments during the studies-you have helped keep me smiling. The Royal Thai Government Scholarship, for financial support throughout my studies. The School of Pharmacy and Life Sciences at The Robert Gordon University, for financial aid during the writing up period. The British Association for Cancer Research (BACR), for the travel grants to attend and present my research in the 2006 and 2007 National Cancer Research Institute (NCRI) Conference in Birmingham. v List of Publications List of Publications Abstract: 1. Warasiha B , Knott R, Boyle S. Investigation of the role of xenobiotic metabolising enzymes in breast cancer development. The Journal of Nutrition. 2004 December; 134 (12 Suppl): 3525S. (Poster presented as part of the International Research Conference on Food, Nutrition and Cancer held in Washington, DC, USA, July, 2004). Conference proceedings: 1. Warasiha B, Knott R, McFadyen M. (2007) Comparative analysis of Cytochrome P450 mRNA Expression between Human Caucasian and Afro-Caribbean Breast Cancer Cell Lines. In the proceedings of National Cancer Research Institute (NCRI) Cancer Conference October 2007 at the ICC in Birmingham, UK. (Oral and poster presentation) 2. Warasiha B, Knott R, McFadyen M. (2007) Differential Cytochrome P450 mRNA Profile between Non-invasive and Invasive Breast Cancer Phenotypes. In the proceedings of National Cancer Research Institute (NCRI) Cancer Conference October 2007 at the ICC in Birmingham, UK. (Poster presentation) 3. Kay G, Cairns D, McCaughan B, Warasiha B. (2007) The Design, Synthesis and Biological Evaluation of Novel Prodrugs for the Treatment of Cystinosis. In the proceedings of British Conference Pharmaceutical (BCP) September 2007 at the Manchester central in Manchester, UK. (Poster presentation) 4. Kay G, Warasiha B, Melville M, Mincher DJ. (2007) Design, Synthesis and Biological Evaluation of Novel DNA Binding Agents. In the proceedings of British Conference Pharmaceutical (BCP) September 2007 at the Manchester central in Manchester, UK. (Poster presentation) 5. Warasiha B, Knott R, McFadyen M. (2006) Profile of cytochrome P450 mRNA expression in oestrogen positive and negative breast cancer cell lines. In the Proceedings of National Cancer Research Institute (NCRI) Cancer Conference October 2006 at the ICC in Birmingham, UK. (Poster presentation) vi Table of Contents Table of Contents Title page-------------------------------------------------------------------------------------- i Abstract --------------------------------------------------------------------------------------- ii Acknowledgements -------------------------------------------------------------------------- iv List of Publications -------------------------------------------------------------------------- vi Table of Contents ----------------------------------------------------------------------------- vii List of Abbreviations ------------------------------------------------------------------------
Recommended publications
  • CYP26C1 Is a Hydroxylase of Multiple Active Retinoids and Interacts with Cellular Retinoic Acid Binding Proteins S
    Supplemental material to this article can be found at: http://molpharm.aspetjournals.org/content/suppl/2018/02/23/mol.117.111039.DC1 1521-0111/93/5/489–503$35.00 https://doi.org/10.1124/mol.117.111039 MOLECULAR PHARMACOLOGY Mol Pharmacol 93:489–503, May 2018 Copyright ª 2018 by The American Society for Pharmacology and Experimental Therapeutics CYP26C1 Is a Hydroxylase of Multiple Active Retinoids and Interacts with Cellular Retinoic Acid Binding Proteins s Guo Zhong, David Ortiz, Alex Zelter, Abhinav Nath, and Nina Isoherranen Departments of Pharmaceutics (G.Z., N.I.) and Medicinal Chemistry (D.O., A.N.), School of Pharmacy, and Department of Biochemistry, School of Medicine (A.Z.), University of Washington, Seattle, Washington Received October 31, 2017; accepted February 22, 2018 ABSTRACT Downloaded from The clearance of retinoic acid (RA) and its metabolites is believed orientation of retinoids within the CYP26C1 active site. In compar- to be regulated by the CYP26 enzymes, but the specific roles of ison with other CYP26 family members, CYP26C1 was up to CYP26A1, CYP26B1, and CYP26C1 in clearing active vitamin A 10-fold more efficient in clearing 4-oxo-atRA (intrinsic clearance metabolites have not been defined. The goal of this study was to 153 ml/min/pmol) than CYP26A1 and CYP26B1, suggesting that establish the substrate specificity of CYP26C1, and determine CYP26C1 may be important in clearing this active retinoid. In whether CYP26C1 interacts with cellular retinoic acid binding support of this, CRABPs delivered 4-oxo-atRA and atRA for proteins (CRABPs). CYP26C1 was found to effectively metabo- metabolism by CYP26C1.
    [Show full text]
  • Impaired Hepatic Drug and Steroid Metabolism in Congenital Adrenal
    European Journal of Endocrinology (2010) 163 919–924 ISSN 0804-4643 CLINICAL STUDY Impaired hepatic drug and steroid metabolism in congenital adrenal hyperplasia due to P450 oxidoreductase deficiency Dorota Tomalik-Scharte1, Dominique Maiter2, Julia Kirchheiner3, Hannah E Ivison, Uwe Fuhr1 and Wiebke Arlt School of Clinical and Experimental Medicine, Centre for Endocrinology, Diabetes and Metabolism (CEDAM), University of Birmingham, Birmingham B15 2TT, UK, 1Department of Pharmacology, University Hospital, University of Cologne, 50931 Cologne, Germany, 2Department of Endocrinology, University Hospital Saint Luc, 1200 Brussels, Belgium and 3Department of Pharmacology of Natural Products and Clinical Pharmacology, University of Ulm, 89019 Ulm, Germany (Correspondence should be addressed to W Arlt; Email: [email protected]) Abstract Objective: Patients with congenital adrenal hyperplasia due to P450 oxidoreductase (POR) deficiency (ORD) present with disordered sex development and glucocorticoid deficiency. This is due to disruption of electron transfer from mutant POR to microsomal cytochrome P450 (CYP) enzymes that play a key role in glucocorticoid and sex steroid synthesis. POR also transfers electrons to all major drug- metabolizing CYP enzymes, including CYP3A4 that inactivates glucocorticoid and oestrogens. However, whether ORD results in impairment of in vivo drug metabolism has never been studied. Design: We studied an adult patient with ORD due to homozygous POR A287P, the most frequent POR mutation in Caucasians, and her clinically unaffected, heterozygous mother. The patient had received standard dose oestrogen replacement from 17 until 37 years of age when it was stopped after she developed breast cancer. Methods: Both subjects underwent in vivo cocktail phenotyping comprising the oral administration of caffeine, tolbutamide, omeprazole, dextromethorphan hydrobromide and midazolam to assess the five major drug-metabolizing CYP enzymes.
    [Show full text]
  • 4Β-Hydroxycholesterol As Biomarker for Variation in CYP3A Activity
    ȕ-Hydroxycholesterol as biomarker for variation in CYP3A activity Dissertation for the Degree of Philosophiae Doctor (Ph.D.) Kristine Hole 2018 Center for Psychopharmacology Diakonhjemmet Hospital Oslo Department of Pharmaceutical Biosciences School of Pharmacy Faculty of Mathematics and Natural Sciences University of Oslo © Kristine Hole, 2018 Series of dissertations submitted to the Faculty of Mathematics and Natural Sciences, University of Oslo No. ISSN 1501-7710 All rights reserved. No part of this publication may be reproduced or transmitted, in any form or by any means, without permission. Cover: Hanne Baadsgaard Utigard. Print production: Reprosentralen, University of Oslo. TABLE OF CONTENTS ACKNOWLEDGEMENTS ...................................................................................................... II LIST OF PUBLICATIONS ..................................................................................................... III ABBREVIATIONS..................................................................................................................IV ABSTRACT.............................................................................................................................. V 1 INTRODUCTION.............................................................................................................. 1 1.1 Variability in drug response ....................................................................................... 1 1.2 Drug metabolism .......................................................................................................
    [Show full text]
  • NIH Public Access Author Manuscript Pharmacogenet Genomics
    NIH Public Access Author Manuscript Pharmacogenet Genomics. Author manuscript; available in PMC 2013 February 01. NIH-PA Author ManuscriptPublished NIH-PA Author Manuscript in final edited NIH-PA Author Manuscript form as: Pharmacogenet Genomics. 2012 February ; 22(2): 159–165. doi:10.1097/FPC.0b013e32834d4962. PharmGKB summary: very important pharmacogene information for cytochrome P450, family 2, subfamily C, polypeptide 19 Stuart A. Scotta, Katrin Sangkuhlc, Alan R. Shuldinere,f, Jean-Sébastien Hulotb,g, Caroline F. Thornc, Russ B. Altmanc,d, and Teri E. Kleinc aDepartment of Genetics and Genomic Sciences bCardiovascular Research Center, Mount Sinai School of Medicine, New York, New York cDepartments of Genetics dBioengineering, Stanford University, Stanford, California eDivision of Endocrinology, Diabetes and Nutrition, University of Maryland School of Medicine fGeriatric Research and Education Clinical Center, Veterans Administration Medical Center, Baltimore, Maryland, USA gDepartment of Pharmacology, Université Pierre et Marie Curie-Paris 6, INSERM UMR S 956, Pitié-Salpêtrière University Hospital, Paris, France Abstract This PharmGKB summary briefly discusses the CYP2C19 gene and current understanding of its function, regulation, and pharmacogenomic relevance. Keywords antidepressants; clopidogrel; CYP2C19*17; CYP2C19*2; CYP2C19; proton pump inhibitors; rs4244285 Introduction The cytochrome P450, family 2, subfamily C, polypeptide 19 (CYP2C19) gene is located within a cluster of cytochrome P450 genes (centromere-CYP2C18-CYP2C19-CYP2C9- CYP2C8-telomere) on chromosome 10q23.33. The CYP2C19 enzyme contributes to the metabolism of a large number of clinically relevant drugs and drug classes such as antidepressants [1], benzodiazepines [2], mephenytoin [3], proton pump inhibitors (PPIs) [4], and the antiplatelet prodrug clopidogrel [5]. Similar to other CYP450 genes, inherited genetic variation in CYP2C19 and its variable hepatic expression contributes to the interindividual phenotypic variability in CYP2C19 substrate metabolism.
    [Show full text]
  • Identification and Developmental Expression of the Full Complement Of
    Goldstone et al. BMC Genomics 2010, 11:643 http://www.biomedcentral.com/1471-2164/11/643 RESEARCH ARTICLE Open Access Identification and developmental expression of the full complement of Cytochrome P450 genes in Zebrafish Jared V Goldstone1, Andrew G McArthur2, Akira Kubota1, Juliano Zanette1,3, Thiago Parente1,4, Maria E Jönsson1,5, David R Nelson6, John J Stegeman1* Abstract Background: Increasing use of zebrafish in drug discovery and mechanistic toxicology demands knowledge of cytochrome P450 (CYP) gene regulation and function. CYP enzymes catalyze oxidative transformation leading to activation or inactivation of many endogenous and exogenous chemicals, with consequences for normal physiology and disease processes. Many CYPs potentially have roles in developmental specification, and many chemicals that cause developmental abnormalities are substrates for CYPs. Here we identify and annotate the full suite of CYP genes in zebrafish, compare these to the human CYP gene complement, and determine the expression of CYP genes during normal development. Results: Zebrafish have a total of 94 CYP genes, distributed among 18 gene families found also in mammals. There are 32 genes in CYP families 5 to 51, most of which are direct orthologs of human CYPs that are involved in endogenous functions including synthesis or inactivation of regulatory molecules. The high degree of sequence similarity suggests conservation of enzyme activities for these CYPs, confirmed in reports for some steroidogenic enzymes (e.g. CYP19, aromatase; CYP11A, P450scc; CYP17, steroid 17a-hydroxylase), and the CYP26 retinoic acid hydroxylases. Complexity is much greater in gene families 1, 2, and 3, which include CYPs prominent in metabolism of drugs and pollutants, as well as of endogenous substrates.
    [Show full text]
  • Methylation Status of Vitamin D Receptor Gene Promoter in Adrenocortical Carcinoma
    UNIVERSITÀ DEGLI STUDI DI PADOVA DEPARTMENT OF CARDIAC, THORACIC AND VASCULAR SCIENCES Ph.D Course Medical Clinical and Experimental Sciences Curriculum Clinical Methodology, Endocrinological, Diabetological and Nephrological Sciences XXIX° SERIES METHYLATION STATUS OF VITAMIN D RECEPTOR GENE PROMOTER IN ADRENOCORTICAL CARCINOMA Coordinator: Ch.mo Prof. Annalisa Angelini Supervisor: Ch.mo Prof. Francesco Fallo Ph.D Student: Andrea Rebellato TABLE OF CONTENTS SUMMARY 3 INTRODUCTION 4 PART 1: ADRENOCORTICAL CARCINOMA 4 1.1 EPIDEMIOLOGY 4 1.2 GENETIC PREDISPOSITION 4 1.3 CLINICAL PRESENTATION 6 1.4 DIAGNOSTIC WORK-UP 7 1.4.1 Biochemistry 7 1.4.2 Imaging 9 1.5 STAGING 10 1.6 PATHOLOGY 11 1.7 MOLECULAR PATHOLOGY 14 1.7.1 DNA content 15 1.7.2 Chromosomal aberrations 15 1.7.3 Differential gene expression 16 1.7.4 DNA methylation 17 1.7.5 microRNAs 18 1.7.6 Gene mutations 19 1.8 PATHOPHYSIOLOGY OF MOLECULAR SIGNALLING 21 PATHWAYS 1.8.1 IGF-mTOR pathway 21 1.8.2 WNTsignalling pathway 22 1.8.3 Vascular endothelial growth factor 23 1.9 THERAPY 24 1.9.1 Surgery 24 1.9.2 Adjuvant Therapy 27 1.9.2.1 Mitotane 27 1.9.2.2 Cytotoxic chemotherapy 30 1.9.2.3 Targeted therapy 31 1.9.2.4 Therapy for hormone excess 31 1.9.2.5 Radiation therapy 32 1.9.2.6 Other local therapies 32 1.10 PROGNOSTIC FACTORS AND PREDICTIVE MARKERS 32 PART 2: VITAMIN D 35 2.1 VITAMIN D AND ITS BIOACTIVATION 35 2.2 THE VITAMIN D RECEPTOR 37 2.3 GENOMIC MECHANISM OF 1,25(OH)2D3-VDR COMPLEX 38 2.4 CLASSICAL ROLES OF VITAMIN D 40 2.4.1 Intestine 40 2.4.2 Kidney 41 2.4.3 Bone 41 2.5 PLEIOTROPIC
    [Show full text]
  • Identification and Characterization of CYP2C18 in the Cynomolgus Macaque (Macaca Fascicularis)
    NOTE Toxicology Identification and Characterization of CYP2C18 in the Cynomolgus Macaque (Macaca fascicularis) Yasuhiro UNO1)*, Kiyomi MATSUNO1), Chika NAKAMURA1), Masahiro UTOH1) and Hiroshi YAMAZAKI2) 1)Pharmacokinetics and Bioanalysis Center (PBC), Shin Nippon Biomedical Laboratories (SNBL), Kainan, Wakayama 642–0017 and 2)Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida, Tokyo 194–8543, Japan (Received 3 August 2009/Accepted 15 September 2009/Published online in J-STAGE 25 November 2009) ABSTRACT. The macaque is widely used for investigation of drug metabolism due to its evolutionary closeness to the human. However, the genetic backgrounds of drug-metabolizing enzymes have not been fully investigated; therefore, identification and characterization of drug-metabolizing enzyme genes are important for understanding drug metabolism in this species. In this study, we isolated and char- acterized a novel cytochrome P450 2C18 (CYP2C18) cDNA in cynomolgus macaques. This cDNA was highly homologous (96%) to human CYP2C18 cDNA. Cynomolgus CYP2C18 was preferentially expressed in the liver and kidney. Moreover, a metabolic assay using cynomolgus CYP2C18 protein heterologously expressed in Escherichia coli revealed its activity toward S-mephenytoin 4’-hydrox- ylation. These results suggest that cynomolgus CYP2C18 could function as a drug-metabolizing enzyme in the liver. KEY WORDS: cloning, CYP2C18, cytochrome P450, liver, monkey. J. Vet. Med. Sci. 72(2): 225–228, 2010 Cytochrome P450 (CYP) is a superfamily of some of the Such species differences also could be explained by func- most important drug-metabolizing enzymes and consists of tional disparity of the orthologous CYPs between the two a large number of subfamilies [14]. In humans, the CYP2C species, such as, if an ortholog to human CYP with low (or subfamilies contain important enzymes that metabolize no) expression and drug-metabolizing activity, for example approximately 20% of all prescribed drugs [3].
    [Show full text]
  • Pharmacogenomic Characterization in Bipolar Spectrum Disorders
    pharmaceutics Review Pharmacogenomic Characterization in Bipolar Spectrum Disorders Stefano Fortinguerra 1,2 , Vincenzo Sorrenti 1,2,3 , Pietro Giusti 2, Morena Zusso 2 and Alessandro Buriani 1,2,* 1 Maria Paola Belloni Center for Personalized Medicine, Data Medica Group (Synlab Limited), 35131 Padova, Italy; [email protected] (S.F.); [email protected] (V.S.) 2 Department of Pharmaceutical & Pharmacological Sciences, University of Padova, 35131 Padova, Italy; [email protected] (P.G.); [email protected] (M.Z.) 3 Bendessere™ Study Center, Solgar Italia Multinutrient S.p.A., 35131 Padova, Italy * Correspondence: [email protected] Received: 25 November 2019; Accepted: 19 December 2019; Published: 21 December 2019 Abstract: The holistic approach of personalized medicine, merging clinical and molecular characteristics to tailor the diagnostic and therapeutic path to each individual, is steadily spreading in clinical practice. Psychiatric disorders represent one of the most difficult diagnostic challenges, given their frequent mixed nature and intrinsic variability, as in bipolar disorders and depression. Patients misdiagnosed as depressed are often initially prescribed serotonergic antidepressants, a treatment that can exacerbate a previously unrecognized bipolar condition. Thanks to the use of the patient’s genomic profile, it is possible to recognize such risk and at the same time characterize specific genetic assets specifically associated with bipolar spectrum disorder, as well as with the individual response to the various therapeutic options. This provides the basis for molecular diagnosis and the definition of pharmacogenomic profiles, thus guiding therapeutic choices and allowing a safer and more effective use of psychotropic drugs. Here, we report the pharmacogenomics state of the art in bipolar disorders and suggest an algorithm for therapeutic regimen choice.
    [Show full text]
  • Simulating the Impact of the Interplay Between Cyp2c19 Polymorphisms and Ethnicity on Response to Clopidogrel, Using Pbpk-Pd Models
    SIMULATING THE IMPACT OF THE INTERPLAY BETWEEN CYP2C19 POLYMORPHISMS AND ETHNICITY ON RESPONSE TO CLOPIDOGREL, USING PBPK-PD MODELS. Manoranjenni Chetty, Khaled Abduljalil. Certara UK, Simcyp Division, Level 2-Acero, 1 Concourse Way, Sheffield, United Kingdom. Background PBPK-PD Model: Lua scripting was used within the Simcyp simulator for the PBPK-PD model. A modified indirect response turnover model6, Clopidogrel is a prodrug that produces its anticoagulant effect after with maximum platelet aggregation (MPA%) as the PD marker was used conversion to Clopi-H4, the active metabolite. Clopi-H4 binds to simulate the response to Clopi-H4. % IPA was calculated as: irreversibly to the platelet P2Y12 adenosine diphosphate (ADP) % IPA = [MPApredose – MPApostdose / MPApredose] * 100% receptor, which inhibits platelet aggregation and reduces platelet Clopi-H4 concentrations from the PBPK model were used as the input to reactivity for the platelet’s life span1. the PD model. Clopidogrel is metabolized by two major metabolic pathways. An Model Performance Verification: The PBPK model was verified by esterase-dependent pathway leads to hydrolysis of clopidogrel into comparison of the predicted and clinically observed pharmacokinetic an inactive carboxylic acid derivative (85–92%) while a cytochrome parameters. P450 (CYP) dependent pathway leads to the formation of its active Following the verification of the performance of the PBPK-PD model, 2,3,4 metabolite (clopi-H4) . CYP2C19, CYP2B6, and CYP1A2 first convert simulations were repeated using healthy Chinese PM and Caucasian PM clopidogrel to the 2-oxo-clopidogrel intermediate, which is then populations. The change in % IPA was compared in the 4 groups to metabolised by esterases (about 50%) or converted to Clopi-H4 by determine the need for dosage adjustments.
    [Show full text]
  • 2016 SOT Enterocytes
    Human Enterocytes: Isolation, Cryopreservation, Characterization, and Application in The Evaluation of Drug-Food Interactions Albert P. Li, Qian Yang, David Ho, Carol J. Loretz, Stephen N. Ring, Kirsten Amaral, Utkarsh Doshi, In Vitro ADMET Laboratories Inc., Columbia, MD and Malden, MA Introduction Results Why Enterocytes Successful Isolation and Comparison of Enterocytes and Hepatocytes in Drug Metabolizing Enzyme Activities (pmol/min/million enterocytes) Drug Gene Expression Substrate • Key cell type for oral bioavailability (as a Cryopreservation from Metabolizing Substrate Metabolite HE3005 HE3006 HE3007 HE3008 HE3009 HE3010 HE3011 HE3014 HE3015 HE3016 HE3020 HE3027 HE3029 Conc. (µM) function of permeability, metabolism, efflux) Multiple Donors Enzyme P450 Isoforms CYP2C9 Diclofenac 25 4-OH Diclofenac 1.68 0.59 0.91 0.46 1.18 1.21 0.03 0.44 2.50 2.05 0.31 2.02 0.86 First pass metabolism before the liver • CYP3A4/5 Midazolam 20 1-OH-Midazolam 2.67 0.13 0.99 0.87 0.72 0.46 0.09 0.40 2.55 0.99 0.49 0.68 0.59 Lot Gender Race Age (Years) -ΔCт -ΔCт Ratio of enterocyte to Gene name 2 enterocytes 2 hepatocytes 7-OH Coumarin UGT 7-OH Coumarin 100 8.38 2.30 3.08 1.80 4.32 2.56 1.01 3.55 7.33 5.71 5.83 3.68 6.55 • Target of drug-interactions with orally co- hepatocytes Glucuronide HE3005 M C 23 Sulfate administered substances CYP1A2 0.0000 0.1333 0.00 7-OH Coumarin 100 7-OH Coumarin Sulfate 8.72 2.04 4.04 1.79 7.78 3.32 1.70 2.66 5.23 4.13 1.84 2.69 3.65 HE3006 M C 47 Transferase CYP2B6 0.0024 1.0304 0.00 • Target of enterotoxicity of ingested
    [Show full text]
  • Synonymous Single Nucleotide Polymorphisms in Human Cytochrome
    DMD Fast Forward. Published on February 9, 2009 as doi:10.1124/dmd.108.026047 DMD #26047 TITLE PAGE: A BIOINFORMATICS APPROACH FOR THE PHENOTYPE PREDICTION OF NON- SYNONYMOUS SINGLE NUCLEOTIDE POLYMORPHISMS IN HUMAN CYTOCHROME P450S LIN-LIN WANG, YONG LI, SHU-FENG ZHOU Department of Nutrition and Food Hygiene, School of Public Health, Peking University, Beijing 100191, P. R. China (LL Wang & Y Li) Discipline of Chinese Medicine, School of Health Sciences, RMIT University, Bundoora, Victoria 3083, Australia (LL Wang & SF Zhou). 1 Copyright 2009 by the American Society for Pharmacology and Experimental Therapeutics. DMD #26047 RUNNING TITLE PAGE: a) Running title: Prediction of phenotype of human CYPs. b) Author for correspondence: A/Prof. Shu-Feng Zhou, MD, PhD Discipline of Chinese Medicine, School of Health Sciences, RMIT University, WHO Collaborating Center for Traditional Medicine, Bundoora, Victoria 3083, Australia. Tel: + 61 3 9925 7794; fax: +61 3 9925 7178. Email: [email protected] c) Number of text pages: 21 Number of tables: 10 Number of figures: 2 Number of references: 40 Number of words in Abstract: 249 Number of words in Introduction: 749 Number of words in Discussion: 1459 d) Non-standard abbreviations: CYP, cytochrome P450; nsSNP, non-synonymous single nucleotide polymorphism. 2 DMD #26047 ABSTRACT Non-synonymous single nucleotide polymorphisms (nsSNPs) in coding regions that can lead to amino acid changes may cause alteration of protein function and account for susceptivity to disease. Identification of deleterious nsSNPs from tolerant nsSNPs is important for characterizing the genetic basis of human disease, assessing individual susceptibility to disease, understanding the pathogenesis of disease, identifying molecular targets for drug treatment and conducting individualized pharmacotherapy.
    [Show full text]
  • Human Cytochrome P450 CYP2A13
    [CANCER RESEARCH 60, 5074–5079, September 15, 2000] Human Cytochrome P450 CYP2A13: Predominant Expression in the Respiratory Tract and Its High Efficiency Metabolic Activation of a Tobacco-specific Carcinogen, 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone1 Ting Su, Ziping Bao, Qing-Yu Zhang, Theresa J. Smith, Jun-Yan Hong,2 and Xinxin Ding2 Wadsworth Center, New York State Department of Health, Albany, New York 12201 [T. S., Q-Y. Z., X. D.]; School of Public Health, State University of New York at Albany, Albany, New York [T. S., X. D.]; and Environmental and Occupational Health Sciences Institute, University of Medicine and Dentistry of New Jersey, Piscataway, New Jersey 08854 [Z. B., T. J. S., J-Y. H.] ABSTRACT However, heterologously expressed CYP2A7 showed no catalytic activity (17, 18). CYP2A13 cDNA has not been isolated previously; The human CYP2A subfamily comprises three genes, CYP2A6, the reported protein sequence was deduced from the predicted coding CYP2A7, and CYP2A13. CYP2A6 is active toward many carcinogens and region of a CYP2A13 genomic clone (1). On the basis of its sequence is the major coumarin 7-hydroxylase and nicotine C-oxidase in the liver, whereas CYP2A7 is not functional. The function of CYP2A13 has not been features that resemble the nonfunctional CYP2A7 and CYP2A6v1 (a characterized. In this study, a CYP2A13 cDNA was prepared by RNA- genetic variant of CYP2A6) proteins, the CYP2A13 protein was PCR from human nasal mucosa and was translated using a baculovirus predicted to be nonfunctional in coumarin 7-hydroxylation (1). Be- expression system. In a reconstituted system, the expressed CYP2A13 was cause the deduced amino acid sequence of CYP2A13 shares a 95.4% more active than CYP2A6 in the metabolic activation of hexamethylphos- identity with that of CYP2A6 (1), antibodies and chemical probes for phoramide, N,N-dimethylaniline, 2؅-methoxyacetophenone, and N-nitro- CYP2A6 may interact with CYP2A13.
    [Show full text]