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ENZYME INHIBITION AND BIOAPPLICATIONS Edited by Rakesh Sharma Enzyme Inhibition and Bioapplications Edited by Rakesh Sharma Published by InTech Janeza Trdine 9, 51000 Rijeka, Croatia Copyright © 2012 InTech All chapters are Open Access distributed under the Creative Commons Attribution 3.0 license, which allows users to download, copy and build upon published articles even for commercial purposes, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. After this work has been published by InTech, authors have the right to republish it, in whole or part, in any publication of which they are the author, and to make other personal use of the work. Any republication, referencing or personal use of the work must explicitly identify the original source. As for readers, this license allows users to download, copy and build upon published chapters even for commercial purposes, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. Notice Statements and opinions expressed in the chapters are these of the individual contributors and not necessarily those of the editors or publisher. No responsibility is accepted for the accuracy of information contained in the published chapters. The publisher assumes no responsibility for any damage or injury to persons or property arising out of the use of any materials, instructions, methods or ideas contained in the book. Publishing Process Manager Anja Filipovic Technical Editor Teodora Smiljanic Cover Designer InTech Design Team First published May, 2012 Printed in Croatia A free online edition of this book is available at www.intechopen.com Additional hard copies can be obtained from [email protected] Enzyme Inhibition and Bioapplications, Edited by Rakesh Sharma p. cm. ISBN 978-953-51-0585-5 Contents Preface IX Section 1 Basic Concepts 1 Chapter 1 Enzyme Inhibition: Mechanisms and Scope 3 Rakesh Sharma Section 2 Applications of Enzyme Inhibition 37 Chapter 2 Cytochrome P450 Enzyme Inhibitors from Nature 39 Simone Badal, Mario Shields and Rupika Delgoda Chapter 3 Pharmacomodulation of Broad Spectrum Matrix Metalloproteinase Inhibitors Towards Regulation of Gelatinases 57 Erika Bourguet, William Hornebeck, Janos Sapi, Alain Jean-Paul Alix and Gautier Moroy Chapter 4 Non-Enzymatic Glycation of Aminotransferases and the Possibilities of Its Modulation 85 Iva Boušová, Lenka Srbová and Jaroslav Dršata Chapter 5 Inhibition of Nitric Oxide Synthase Gene Expression: In vivo Imaging Approaches of Nitric Oxide with Multimodal Imaging 115 Rakesh Sharma Chapter 6 Transcriptional Bursting in the Tryptophan Operon of E. coli and Its Effect on the System Stochastic Dynamics 179 Emanuel Salazar-Cavazos and Moisés Santillán Chapter 7 Mechanisms of Hepatocellular Dysfunction and Regeneration: Enzyme Inhibition by Nitroimidazole and Human Liver Regeneration 195 Rakesh Sharma VI Contents Chapter 8 Reversible Inhibition of Tyrosine Protein Phosphatases by Redox Reactions 253 Daniela Cosentino-Gomes and José Roberto Meyer-Fernandes Chapter 9 Feasible Novozym 435-Catalyzed Process to Fatty Acid Methyl Ester Production from Waste Frying Oil: Role of Lipase Inhibition 277 Laura Azócar, Gustavo Ciudad, Robinson Muñoz, David Jeison, Claudio Toro and Rodrigo Navia Chapter 10 Urease Inhibition 303 Muhammad Raza Shah and Zahid Hussain Soomro Preface Enzyme is a protein molecule exhibiting specific activity and binding affinity with its substrate molecule to complete enzyme reaction or biocatalytic reaction. Substrate analogues can inhibit the enzyme reaction and act as enzyme inhibitor. Enzyme inhibition (Enz-ai-m ie-ni-hi-bi-son) means reducing or blocking an enzyme action on specific location of enzyme active site by specific substrate or analogue so called enzyme inhibitor. In modern times, most of the pharmaceutical as well as nutriceutical compounds are marketed as enzyme inhibitors and such inhibitors exhibit their specific action in enzyme inhibition inside cells, bacteria, virus, animal plants and human body. The action of enzyme inhibitors in drug discovery has become a fundamental approach to pharmacology at any pharmaceutical industry, university research lab or drug research center. The present issue has been compiled from various data sources with aim of incorporating a wide range of basic concepts and applied enzyme inhibition evaluation methods in drug discovery. It is aimed at those who are embarking on drug discovery research projects, immobilized enzyme solid state devices as well as relatively experienced pharmacologists, biochemists and pharmacy scientists who might wish to develop their experiments further to the advanced level. While it is not possible to detail and include every possible technique related with enzyme inhibition evaluation in drug design by using specific inhibitors at specific metabolic mechanism(s), the present issue attempts to provide working tips with examples and analysis relevant to a wide range of more commonly available enzyme inhibition techniques. The methods and concepts described in this book are aimed at giving the reader a glimpse of some existing enzyme inhibition studies and also methods with context of each enzyme inhibition method applied for, as well as providing some basis of familiarizing oneself with these biochemical methods. While enzyme inhibition has been used as major approach in drug design in the research and industry over last two decades, it was only later part of 20th century that it has become a major part of so many applications in biochemical engineering, biomedical engineering of miniatured clinical chemistry devices in microbiological, bacteriological, immunological, hormonal testing, nanotechnology, physiological monitoring in health science, plant science and environment research work. This is, at least in part, due to the continued development of new solid state polymer platform, pure enzymes and specific X Preface inhibitors available, better understanding of enzyme inhibition mechanisms and precise detection methods, new awareness of drug discovery and design, with scanning and monitoring accessories. Thanks to the continued joint efforts of governmental, industrial and academic institutions globally to promote the need of new generations of drugs or enzyme inhibitors and new mechanisms of enzyme inhibition. Regardless of the enzymes or drugs and their brands that are used, one should always be able to understand and justify the use of right inhibitor or drug action on enzyme of right choice to drug design for specific study. With this aim, different approaches of enzyme inhibition methods are presented in separate chapters on the use of different enzyme inhibitors. For learners, basic concepts, mechanistic issues, limitations in drug testing, skepticism in enzyme inhibition approach and drug variability due to non-specific analogues of enzyme inhibitors or substrates is presented with a working enzyme inhibition protocols for drug design and analysis of their inhibitory action. In chapter 1, the authors have introduced the basic concepts on enzyme, enzyme reaction, inhibitors and types of inhibition with a handful established applications in drug discovery, immobilized enzyme engineering, and biosensing. Major three basic types of enzyme inhibition kinetics is highlighted for beginners. Examples of substrate analogues and their enzyme inhibition behaviour are illustrated with color schemes. Application of immobilized enzyme on chips for environment monitoring and biosensor development is quite intriguing for engineers, scientists and industrialists. In chapter 2, authors overviewed isolates from Pepermia amplexicaulis and Spathelia sorbifolia plants have examined for CYP inhibitions to exhibit antiprotozoal, chemopreventive and anti-cancer activity. Other plant sources of tea and fruits of Rhytidophyllum tomentosa, Psidium guajava, Symphytium officinale, Momordica charantia showed inhibition properties of these teas against a panel of CYP450 enzymes in order to assess the potential for drug interactions with co-medicated pharmaceuticals. The chapter highlights the potential of a few natural products emanating from the Caribbean: chromene amides isolated from Amyris plumieri, quassinoids isolated from Picrasma excelsa, anhydrosorbifolin isolated from Spathelia sorbifolia and 5-Hydroxy-2,7- dimethyl-8-(3-methyl-but-2-enyl)-2-(4-methyl-penta-1,3-dienyl)-chroman-6-carboxylic acid isolated from Peperomia amplexicaulis. New information is presented on bioactive screens against CYP enzymes in the presence of five aqueous infusions of popularly used herbs; Rhytidophyllum tomentosa, Psidium guajava, Symphytium officinale, Momordica charantia and Picrasma excelsa. Authors emphasized search of CYPs 1A1 and 1B1 activity inhibitors as chemoprotectors such as CA1 and quassin for CYP 1A1; and anhydrosorbifolin and 5-Hydroxy-2,7-dimethyl-8-(3-methyl-but-2-enyl)-2-(4-methyl- penta-1,3-dienyl)-chroman-6-carboxylic acid for CYP 1B1 in search of safer herbal remedies in co-adiministration of medicines in cancer prevention. In chapter 3, authors described pharmacomodulation in metalloprotease enzymes by inhibitors. Authors aim to achieve better metallloprotease (MMP) enzyme inhibitors with good MMP-2 selectivities to increase hydrophobicity and rigidity with the dehydro and didehydro analogues and synthesized (analogue 2a-d and 3a-h). Authors displayed sevral Preface XI examples of inhibitors including pharmacomodulation of galardin®, a powerful broad spectrum MMPI, hydrazide and sulfonylhydrazide-type functions as potential Zinc Binding Compounds for gelatinase enzyme inhibition, double-headed elastase-MMP inhibitor (d-hEMI) able to block elastase and MMP activities, with display of structural-functional models in the last sections. Authors speculate to develop hybrid nanoprobes build from MMPI and fluorescent nanocrystal quantum dots (QDs). Design and chemical synthesis of derivatives of galardin®, selective inhibitors of MMP- 2, tagging with QDs. In hope of photo- and chemical stability of QDs, it is possible that apporoach will enable long-term spatiotemporal tracking of the process of inhibition of MMP-2 enzymes to offer better understanding of physiological process of invasion of melanoma. In chapter 4, authors introduce the readers with nonenzymatic glycation of aminotransferases and modulation of these enzymes as model protein. Further authors emphasized the significance of advanced glycation end-products with introduction of structure, targeting with aim to evaluate potential antiglycation activity of two mitochondrial antioxidants, α-phenyl N-tert-butyl nitrone (PBN) and N- tert-butyl hydroxylamine (NtBHA). In next section, authors established the effect of natural compound on fructose induced AST glycation, basic techniques to determine primary amino groups, enzyme molecular charge of AST modulated by fructose by electrophoresis. Authors attempted to search the compounds with antioxidant and potential antiglycating activities in an illustrated description with a perspective of their use as remedies against diabetic complications. In chapter 5, author introduces the readers with basic mechanism of reduced nitric oxide synthase gene expression and applications of nitric oxide synthase (NOS) inhibition and approaches of nitric oxide (NO) content by using less known multimodal imaging. Nitric oxide imaging techniques utilize mapping NO in tissue using NO specific imaging contrast agents sensitive to fluorescence, magnetic resonance and electron spin resonance. A handful account is presented on NOS expression inhibitors, possibility of dithiacarbamates, paramagnetic complexes for bioimaging of NO. For learners, MRI protocol is given as example. In the light of recent developments in multimodal bioimaging of NO/NOS expression by bioluminescence, fluorescence techniques, a handful information is given in cells, animals, plants and humans body in different diseases including endothelial cell injury, apoptosis, renal, liver, lung, muscle, brain, inflammation, bones, retina with emphasis on multimodal techniques of calcium, ion channels, iron bound complexes. Author speculated the future possibility of NO/NOS bioimaging by combined radioimaging techniques and it remains to see if real-time imaging becomes routine modality. In chapter 6, transcriptional bursting in E.coli tryptophan operon is introduced to explain stochastic dynamics of the tryptophan synthase enzyme feedback inhibition regulatory mechanism at various levels of Trp operon genes. Authors have described in chapter various components of trp operon structure, model development, parameter estimation, and numerical methods with results on stochastic stimulations to evidence the least response time with inhibition-less strain. Authors further investigated that the repressor-operator interaction stimulates transcriptional bursting which shows several dynamic effects on transcriptional bursting possibly by a feedback enzyme-inhibition regulatory mechanism. In chapter 7, a hepatocellular