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The Design and Synthesis of Autotaxin Inhibitors and Analogs of Lysophosphatidic Acid

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The Design and Synthesis of Autotaxin Inhibitors and Analogs of Lysophosphatidic Acid University of Tennessee Health Science Center UTHSC Digital Commons Theses and Dissertations (ETD) College of Graduate Health Sciences 5-2011 The esiD gn and Synthesis of Autotaxin Inhibitors and Analogs of Lysophosphatidic Acid Renuka Niranjan Gupte University of Tennessee Health Science Center Follow this and additional works at: https://dc.uthsc.edu/dissertations Part of the Chemicals and Drugs Commons, and the Pharmaceutics and Drug Design Commons Recommended Citation Gupte, Renuka Niranjan , "The eD sign and Synthesis of Autotaxin Inhibitors and Analogs of Lysophosphatidic Acid" (2011). Theses and Dissertations (ETD). Paper 112. http://dx.doi.org/10.21007/etd.cghs.2011.0121. This Dissertation is brought to you for free and open access by the College of Graduate Health Sciences at UTHSC Digital Commons. It has been accepted for inclusion in Theses and Dissertations (ETD) by an authorized administrator of UTHSC Digital Commons. For more information, please contact [email protected]. The esiD gn and Synthesis of Autotaxin Inhibitors and Analogs of Lysophosphatidic Acid Document Type Dissertation Degree Name Doctor of Philosophy (PhD) Program Pharmaceutical Sciences Research Advisor Duane D. Miller, Ph.D. Committee Isaac O. Donkor, Ph.D. Richard E. Lee, Ph.D. Wei Li, Ph.D. Gabor J. Tigyi, M.D., Ph.D. DOI 10.21007/etd.cghs.2011.0121 Comments Two year embargo expired May 2013 This dissertation is available at UTHSC Digital Commons: https://dc.uthsc.edu/dissertations/112 THE DESIGN AND SYNTHESIS OF AUTOTAXIN INHIBITORS AND ANALOGS OF LYSOPHOSPHATIDIC ACID A Dissertation Presented for The Graduate Studies Council The University of Tennessee Health Science Center In Partial Fulfillment Of the Requirements for the Degree Doctor of Philosophy From The University of Tennessee By Renuka Niranjan Gupte May 2011 Copyright © 2011 by Renuka N. Gupte All rights reserved ii Dedication This dissertation is dedicated to my greatest supporters and idols, my husband and friend Dr.Varun Vaidya and my parents, Mr.Niranjan Gupte and Ms.Savita Gupte. iii Acknowledgements I would like to thank my major professor and mentor Dr. Duane D. Miller for giving me a chance to advance professionally and for his support and help throughout this dissertation process. I would also like to thank my dissertation committee members; Dr. Gabor J. Tigyi, Dr. Wei Li, Dr. Isaac O. Donkor and Dr. Richard E. Lee for providing me excellent feedback and also for their continuous encouragement. Without them this dissertation could not have been possible. I give special thanks to Dr.Yan Lu, Dr.Anjaiah Siddam, Dr. Jayprakash Pagadala, Dr. Zhao Wang and Dr.James Fells for their expertise, support, assistance and friendship. I would also like to thank all the members of the “Miller research group”, Dr. Charlie Duke, Dr. Renukadevi Patil, Dr.Shivaputra Patil, Dr. Kui Zeng and Mr.Jianjun Chen. I thank all the members of Dr.Tigyis Lab, especially Dr.Yuko Fujiwara and Dr.Dianna Liu. I would also like to extend thanks to our collaborators on the research projects in this dissertation: Dr. Daniel Baker (University of Memphis) and Dr. Abby Parrill (University of Memphis) and their lab members. I would also like to thank Dr. Paul W.Erhardt for allowing me to complete the synthesis of the last few compounds at the University of Toledo. I’d like to thank my family who supported me and kept me encouraged. The research on autotaxin inhibitors (Chapters 2 and 3) was supported by NIH grant CA92160 (Dr.Tigyi), the Van Vleet Oncology Research Fund (Dr.Tigyi), the Breast Cancer Research Foundation (Dr.Panupinthu) and Lpath Inc. (Dr.Mills). iv Abstract Lysophosphatidic acid (LPA) is a naturally occurring bioactive phospholipid. It has a wide array of biological effects like cell proliferation, survival, migration, apoptosis invasion, wound healing and angiogenesis. Autotaxin (ATX) was identified as an autocrine tumor cell motility factor from A2058 melanoma conditioned medium. ATX has lysophospholipase D enzyme activity and is responsible for the cleavage of lysophophatidylcholine (LPC) leading to the generation of LPA. Antagonists of Autotaxin would have a potential therapeutic application in cancer research. Chapter 1 is an introduction of LPA and autotaxin. It provides the background and significance of the research. Chapter 2 explores the synthesis of the stereoisomers of 3-CCPA and the pharmacological activites of these isomers. Chapter 3 discusses the design and synthesis of benzyl and naphthalene-methyl phosphonic acid inhibitors of autotaxin with anti-invasive and anti-metastatic actions. Chapter 4 elaborates on the search for new non-lipid, drug like LPA analogs. Chapter 5 provides an overview of the work detailed in the dissertation; as well as future directions that will help further the scope of these projects. v Table of Contents Chapter 1. Introduction ....................................................................................................1 Biosynthesis and Catabolism of LPA .............................................................................1 Extracellular Generation of LPA ....................................................................................2 The Lyso-PLD/Autotaxin Pathway ...........................................................................2 The Phospholipase A Pathway ..................................................................................2 Mild Oxidation of Low Density Lipoprotein ............................................................2 Intracellular Generation of LPA......................................................................................3 The Glycerophosphate Acyl Transferase (GPAT) Pathway .....................................3 The Action of Acyl Glycerol Kinase .........................................................................3 Reduction of Acyl Dihydroxy Acetone Phosphate ...................................................3 LPA Degradation ............................................................................................................3 LPA Degradation by Phosphatases ...........................................................................3 LPA Degradation by Lysophospholipases ................................................................4 LPA Degradation by Acyltranferases........................................................................4 LPA Receptors ................................................................................................................4 LPA Plasma Membrane GPCR .................................................................................4 PPARγ: An Intracellular Receptor of LPA ...............................................................5 Importance of LPA in Physiological Systems ................................................................5 LPA in Malignant Neoplasm .....................................................................................5 LPA in Nervous System ............................................................................................6 LPA in Cardiovascular System .................................................................................6 LPA in Organ Fibrosis ..............................................................................................6 LPA in Bone Metabolism ..........................................................................................6 LPA in Immunity.......................................................................................................7 Physiological and Pathological Role of Autotaxin .........................................................7 Chapter 2. Synthesis and Pharmacological Evaluation of the Stereoisomers of 3-Carba Cyclic-Phosphatidic Acid .................................................................................10 Biology ..........................................................................................................................12 Chemistry ......................................................................................................................15 Conclusion ....................................................................................................................19 Chapter 3. Benzyl and Naphthalene-Methyl Phosphonic Acid Inhibitors of Autotaxin with Anti-Invasive and Anti-Metastatic Actions ........................................20 Chemical Synthesis .......................................................................................................21 Biological Results .........................................................................................................23 Effect of 4-Substituted Benzyl Phosphonic Acid and 6-Substituted Naphthalene-2yl-Methyl Phosphonic Acid Derivatives on Autotaxin and LPAR ..............................................................................................................23 ATX Inhibitors Block Hepatocarcinoma Invasion of Mesothelial and HUVEC Monolayers .....................................................................................................33 ATX Inhibitors Reduce B16 Melanoma Metastasis In Vivo ..................................33 vi Experimental Section ....................................................................................................37 Chemistry ................................................................................................................37 Biology ....................................................................................................................44 Autotaxin Inhibition
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