INFORMATION TO USERS This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter free, while others may be from any type of computer printer. The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. Oversize materials (e.g., maps, drawings, charts) are reproduced by sectioning the original, beginning at the upper left-hand comer and continuing from left to right in equal sections with small overlaps. Each original is also photographed in one exposure and is included in reduced form at the back of the book. Photographs included in the original manuscript have been reproduced xerographically in this copy. Higher quality 6” x 9” black and white photographic prints are available for any photographs or illustrations appearing in this copy for an additional charge. Contact UMI directly to order. UMI A Bell & Howell Information Company 300 North Zeeb Road, Ann Arbor MI 48106-1346 USA 313/761-4700 800/521-0600 DEVELOPMENT OF STEROIDAL INHIBITORS OF CYTOCHROME P450-DEPENDENT ANDROGEN AND ESTROGEN BIOSYNTHESIS DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Patrick John Ward, A.B. ***** The Ohio State University 1996 Dissertation Committee: Approved by R. W. Brueggemeier L. W. Robertson J. V. Hines Advisor Y.C.Iin College of Pharmacy UMI NUmber: 9630999 UMI Microform 9630999 Copyright 1996, by UMI Company. All rights reserved. This microform edition is protected against unauthorized copying under Title 17, United States Code. UMI 300 North Zeeb Road Ann Arbor, MI 48103 First and last, this work is dedicated to my love and my inspiration, my wife Jacqueline. I am forever grateful to my family, the Ward's and the Ebner's, without whose support and love I never would have finished this work. When I began this journey, I never dreamt that it would come to an end. As my graduate career draws to a close, I have come to realize that it is never supposed to end - that the point of the journey is not to arrive. It is with humility, and immense pride that for the remainder of my journey, when asked who I am, I may now answer, "I am a scientist." ACKNOWLEDGEMENTS Most people of my generation do not have heroes. Somehow it isn’t thought proper anymore. My hero is Woody Hayes, and I would like to respectfully acknowledge the inspiration he has provided me in life. I am fortunate to have had many mentors. I would like to recognize the following people as having been such to me. Dr. Robert Breitenbach, Dr. Dean Metter, and Dr. Roger DeRoos -- gentlemen, your words and their knowledge ring in my ears to this day. Dr. John Neff, Dr. Thomas O’Dorisio, and Dr. Sue ODorisio --1 hope that one day I may follow your example. I would like to express my deep and sincere gratitude to my advisor, Dr. Robert Brueggemeier. His guidance, encouragement and support were of such fundamental importance that I must state that I would not have been capable of completing my graduate studies without them. I would like to thank and acknowledge my committee, Dr. Young Lin, Dr. Larry Robertson and Dr. Jennifer Hines. Their guidance and insight were invaluable. I owe a large debt of gratitude to my colleagues in the laboratory, Dr. Carl Lovely, Dr. Jill O’Reilly, Dr. Mike Darby, Muriel Liberto, Anne Quinn, Abijit Bhat, and Damon Sharp. Their assistance, knowledge and insight were invaluable. I would like to thank and acknowledge Dr. Albert Soloway, Dr. Robert Curley, Dr. Raymond Doskotch, Dr. Duane Miller, Dr. Dennis Feller, Dr. Nigel Preistley, and Dr. Steve Bergmeier for their knowledge and insight. Teaching goes far beyond the simple dissemination of information. These men understand this implicitly. I would like to acknowledge those who went before me and illuminated the path, Dr. Mustapha Beleh, Dr. Soheila Ebrahemian, Dr. Mike Pcolinsky, Dr. Jeff Cristoff, and Dr. Jill O’Reilly. Finally, it is with great sadness that I would like to remember two individuals whose stay on this earth ended while I did this work, Dean Thomas Harris and my Mend Herman. v ...pay no attention to the man behind the curtain. -The Wizard of Oz Life is too important to be taken too seriously. -Oscar Wilde We must not be suspicious of the answers nature provides us, for she is always truthful. Rather we should be careful of the questions we ask of her. -A Szent-Georgi VITA August 9, 1963 .............................................. Born, Columbus, Ohio 1986 ...............................................................A.B., University of Missouri, Columbia, Missouri 1987-Present ................................................. Medical Scientist Training Program Fellow, The Ohio State University, Columbus, Ohio FIELDS OF STUDY Major Field: Medicinal Chemistry Studies in the biochemistry and endocrinology of steroidal inhibitors of steroid biosynthesis for the purpose of developing effective therapies for the treatment of steroid-dependent tumors of the breast and prostate. TABLE OF CONTENTS Page DEDICATION.........................................................................................ii ACKNOWLEDGMENTS....................................................................... iv VITA.......................................................................................................vii LIST OF FIGURES..................................................................................xi LIST OF TABLES....................................................................................xv LIST OF SCHEMES.............................................................................. xvi ABSTRACT............................................................................................ xvii CHAPTER 1 INTRODUCTION...................................................................................1 1.1 Cancer. ............................................................................................... 1 1.2 Prostate Cancer ..................................................................................4 1.2.1 History..................................................................................... 4 1.2.2 Incidence, Mortality, Prevalence a & Risk ............................... 4 1.2.3 Screening ..................................................................................5 1.2.4 Anatomy & Histology ...............................................................6 1.2.5 Diagnosis & Tumor Grading .................................................... 8 1.2.6 Clinical Staging & Prognosis ................................................... 8 1.2.7 Carcinoma of the Prostate vs. Benign Prostatic Hypertrophy ................................................... 9 1.2.8 Pathophysiology of Disease Progression .................................. 12 1.2.8.1 Androgen Receptor Levels ..............................................12 1.2.8.2 Androgen Receptor Mutations .......................................13 1.2.8.3 Intraame/Autocrine/Paracrine Factors ...........................17 1.2.8.3.1 Growth Factors ......................................................17 viii 1.2.8.3.2 Other Epithelial-Stromal Interactions .................27 1.2.8.3.3 Other Endocrine Factors ...................................... 28 1.2.8.4 Genetic Alterations: Oncogenes, Tumor Suppressor Genes, & Chromosomal Alterations .......... 32 1.2.8.5 Adaptation vs. Selection .................................................. 34 1.3 Prostate Cancer Treatment Modalities ...........................................34 1.3.1 Surgery, Radiation & Cytotoxic Chemotherapy .......................34 1.3.2 Endocrine Manipulation .......................................................... 35 1.3.2.1 Hypothalamic-Pituitary-Testicular Axis Disruption ............................................................... 36 1.3.2.1.1 Orchiectomy & Hypophysectomy ............................36 1.3.2.1.2 Estrogens ..............................................................37 1.3.2.1.3 Progestins ...............................................................38 1.3.2.1.4 GnRH Agonists .................................................... 38 1.3.2.2 Antiandrogens ................................................................. 41 1.3.2.2.1 Nonsteroidal Antiandrogens .................................. 42 1.3.2.2.2 Steroidal Antiandrogens ........................................ 43 1.3.2.3 Disruption of Androgen Biosynthesis ............................47 1.3.2.3.1 5a-Reductase..........................................................48 1.3.2.3.2 17,20-Lyase............................................................ 54 1.4 Breast Cancer ...................................................................................71 1.4.1 Breast Cancer Treatment ..........................................................72 1.4.1.1 Hypothalamic-Pituitary-Ovarian Axis Disruption 74 1.4.1.1.1 Oophorectomy, Hypophysectomy & Adrenalectomy ....................................................74 1.4.1.1.2 Progestins ...............................................................75