INFORMATION TO USERS This manuscript has been reproduced from the microfihn master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be from any type o f 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 Infomiation Company 300 North Zed) Road, Ann Arbor MI 48106-1346 USA 313/761-4700 800/521-0600 i j DEVELOPMENT OF DRUGS FOR HUMAN PROSTATE CANCER DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Chiung-Tong Chen, M.S. ***** The Ohio State University 1997 Dissertation Committee: Approved by Dr. M. Guillaume Wientjes, Adviser Dr. Jessie L.-S. Au Dr. William L. Hayton Adviser Dr. Kenneth K. Chan College of Pharmacy UMI Number: 9813235 UMI Microform 9813235 Copyright 1998, 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, Ml 48103 ABSTRACT Prostate cancer represents a growing public health problem in the United States and Europe. In 1997, an estimate of 43% of total newly diagnosed cancers (334.500 new cases) in men were adenocarcinomas of the prostate, the most frequent malignancy in men currently. Fifteen to twenty-five percent of newly diagnosed cases had been reported as advanced stages o f diseases. The benefit o f survival from currently available treatment is minimal. It is projected that 41,800 men will die of prostate cancer in the United States in 1997. Progress in the understanding of prostate tumor biology and the development of effective therapy has been hampered by lack of clinically relevant tumor models. The work presented in this dissertation focused on the development of clinically relevant prostate tumor models for drug activity evaluation, and on the evaluation of pharmacologic effects of two anticancer agents in human prostate tumors. The major conclusions and contributions of the research in this dissertation are as follows: (a) The establishment of pharmacodynamics of paclitaxel (Chapter 2) and doxorubicin (Chapter 4) in histocultures of patient and three human xenograft tumors (Chapter 7). These data indicate that paclitaxel and doxorubicin are active against early stage and androgen dependent prostate cancer, with the antiproliferative effect being more prominent than the cytotoxic (i.e., cell kill) effect. It is noted that these results are the first to delineate the two effects separately, (b) The apoptotic effect of paclitaxel in patient tumors in their native 3-dimensional state is cell cycle specific (Chapter 3). These results confirm the previously reported cell cycle specificity of paclitaxel-induced apoptosis in monolayer cultures o f human cancer cells, and indicate that the microenvironment of solid tumors do not alter this property of paclitaxel effect, (c) the concentrations of doxorubicin delivered to prostate tissue in dogs, after an intravenous injection, are insufiBcient to deliver the concentrations needed to produce >50% antiproliferation and cytotoxicity, respectively, in patient tumors (Chapters 5 and 6). These results demonstrate that insufficient drug delivery to tumors is one cause of the lack of clinical response, and have led to the development of regional therapy where the drug is delivered in high concentrations directly to the prostate, (d) The CWR22, CWR22R and CWR91 human prostate xenograft tumors show pharmacodynamics of paclitaxel and doxorubicin that are qualitatively and quantitatively similar to those in patient tumors (Chapter 7). It is noted that these models are the first to show responses to drugs similar to patient tumors; other models such as monolayer cultures of human cancer cells overpredict chemosensitivity by >10 fold. These results establish the three xenograft tumors as clinically relevant models for drug activity evaluation. As discussed in Chapter 8, Conclusions and Perspectives, the results described in this dissertation has led to several avenues of research in our laboratory that focus on the development of new effective treatments for prostate cancer, a deadly disease with few treatment options. Ill Dedicated to my parents and families IV ACKNOWLEDGMENTS I would like to express my most sincere gratitude to my adviser Dr. Wientjes for his considerate and supportiveness throughout this study. The completion of this study encompasses 5 years of my efforts at The Ohio State University. With as much sincerity, I am grateful to Dr. Au for the her advises and her intimate collaboration with Dr. Wientjes, from which intellectual interactions have been built upon and critical scientific demanding drove me toward being mature mind of sciences. I wish to thank my advisory committee members, Drs. William L. Hayton and Kenneth K. Chan, for helpful advisory discussions. I want to thank Dr. Gan, Y. and Lu, J. for technical support and sharing study materials and results; Drs. Yen, J., Kuh, H-J., Kuan, H.Y., and Johnson, A. and Millenbaugh, N., and Jang, S.H. for stimulating discussions; Dr. Koo, P., Johnston, J., Kellough, D A., Jones, W., Jurcisek, J., Lembach, M., Koolemans-Beynen, A., and Cai, J. for technical support. Friendships from others in the laboratory and the college have been appreciated. Special thanks to my parents, parents-in-law and families for their long-term supports in every way. Supports from my wife and the duty as a father have been the motives and joys that help me through. VITA December 4, 1962 ......................................... Bom - Taiwan, Republic of China 1986 ................................................................. B.S. in Pharmacy, China Medical College 1988 ................................................................. M.S. in Pharmacology, National Yang-Ming Medical College 1990- 1991....................................................... Vice Researcher, Shu-Lin Research Center, China Chemical & Pharmaceutical Group 1990 - 1991....................................................... Registered Pharmacist 1992 ................................................................. Assistant Research Fellow, Institute of Biological Chemistry, Academia Sinica 1992 - present ................................................. Graduate Research Associate, The Ohio State University PUBLICATIONS Papers 1. Chen, C.T., Au, J.L.-S., Gan, Y.B., and Wientjes, M.G. Differential time dependency of antiproliferative and apoptotic effects of taxol in human prostate tumors. Urol. Oncol., 3: 11-17, 1997. 2. Ho, C.L., Lin, Y.L., Chen, W.C., Yu, H.M., Wang, K.T., Hwang, L.L., and Chen, C.T. Immunogenicity of mastoparan B, a cationic tetradecapeptide isolated from the hornet (Vespa basalis) venom, and its structural requirements. Toxicon, VI 33:1443-1451, 1995. 3. Ho, C.L., Hwang, L.L., Lin, Y.L., Chen, C.T., Yu, H.M., and Wang, K.T. Cardiovascular effects of mastoparan B and its structural requirements. Eur. J. Pharmacol., 259:259-264, 1994. 4. Ho, C.L., Hwang, L.L., and Chen, C.T. Edema-inducing activity of a lethal protein with phospholipase A, activity isolated from the black-bellied hornet (Vespa basalis) Venom. Toxicon, 31:605-613, 1993. 5. Chen, C.T., Chan, J.Y.H., Barnes, C.D., and Chan, S.H.H. Tonic suppression of baroreceptor reflex by endogenous neurotensin in the rat. Regul. Peptides, 28:23-37, 1990. Abstracts 1. Gan, Y., Au, J.L.-S., Chen, C.T., and Wientjes, M.G. Cytotoxicity of suramin, geldanamycin, cytochalasin E, and tfiiacetazone in human prostate cancer xenografts. Proc. Am. Assoc. Cancer Res., 38:219, 1997. 2. Au, J.L.-S., Li, D., Gao, X., Gan, Y , Chen, C.T., Ge, S., Johnson, A.L., and Wientjes, M.G. Time-dependent taxol cytotoxicity. Proc. Am. Assoc. Cancer Res., 38:4, 1997. 3. Au, J.L.-S., Li, D., Gao, X., Gan, Y , Chen, C.T., Yen, W.C., Johnson, A.L., and Wientjes, M.G. Time-dependent taxol effect in human and rat solid tumors. Archives of Otolaryngology-Head and Neck Surgery, 4th International Conference on Head and Neck Cancer, 493:193, 1996. 4. Chen, C.T., Gan, Y.B., Au, J.L-S., and Wientjes, M.G. Response of human prostate tumors to taxol: Relationship with expression of multidrug resistance p- glycoprotein and bcl-2. Proc. Am. Assoc. Cancer Res., 37:368, 1996. 5. Weaver, J R., Kalns, J.E., Schmittgen, T.D., Chen, C.T., Lim, C., and Au, J.L.-S. Molecular pharmacodynamic endpoints for antiproliferative agents. Proc. Am. Assoc. Cancer Res., 36:360, 1995. vn 6. Gan, Y.B., Balturshot, G., Millenbaugh, N.E., Chen, C.T., Kalns, J.E., Lim, C , and Wientjes, M.G., Au, J.L.-S. P-glycoprotein expression in human tumors and its correlation with chemosensitivity to taxol and cell mobility. Proc. Am. Assoc. Cancer Res., 36:334, 1995. 7. Chen, C.T., Au, J.L-S., Burgers, J.K., and Wientjes, M.G. Comparative activity of taxol and doxorubicin in human prostate tumors. Proc. Am. Assoc. Cancer Res. 36:320, 1995. 8. Chen, C.T., Au, J.L-S., Burgers, J.K., and Wientjes, M.G. Pharmacodynamics of taxol in human prostate tumors.