Thesis (Ph. D.)--Ohio State University, 2003. / Title from first page of PDF file. Document formatted into pages; contains xiii, 266 p.: ill. Includes abstract and vita. Advisor: Robert S. Coleman, Dept. of Chemistry. Includes bibliographical references (p. 208-222).

A series of studies in mice were performed to determine the interaction of two sulfur nucleophiles, oral n-acetylcysteine (NAC) and intravenous sodium thiosulfate (Na₂S₂O₃) with the anticancer drug mitomycin C (MMC). Neither nucleophile reduced MMC lethality or hematopoietic toxicity. Both increased the antitumor activity of MMC in mice bearing P-388 and L-1210 leukemias. There was no nucleophile reduction of MMC effects on normal bone marrow stem cells (CFUs) using a murine spleen colony forming assay. In contrast, the nucleophiles significantly enhanced MMC bone marrow toxicity. Three clonogenic human tumor cell lines (HEC-1A endometrial, 8226 myeloma, WiDr colon) were relatively resistant to MMC and the nucleophiles did not increase activity. A human breast cancer cell line (MCF-7) was sensitive to MMC and this activity was blocked by glutathione. Oxygen free radical scavengers did not reduce MMC activity. A novel isocratic high performance liquid chromatography (HPLC) assay (48:52, methanol:0.01M phosphate buffer) using ultraviolet detection at 365 nm was used to characterize MMC-protein binding and murine pharmacokinetics. The k' for MMC was 7.91 and 9.86 for porfiromycin. Peaks were confirmed by mass spectroscopy. MMC was bound 30% to albumin and S-9 microsomal proteins and 60-70% to calf thymus DNA. MMC uptake into mouse bone marrow was enhanced by the nucleophiles and was rapidly cleared from the plasma (half-life 0.5 hours). In vitro MMC metabolism with rat liver S-9 microsomes demonstrated production of a polar eluting, putative MMC metabolite (K' = 4.486, lambda maximum 300 nm). This metabolite was inactive in the in vitro clonogenic tumor cell assay. Finally, molecular pharmacology studies using alkaline DNA elution showed that MMC causes both DNA-DNA and DNA-protein crosslinks. There was no evidence for free radical-induced DNA strand scission by MMC. There was also some evidence of moderate DNA protection with the sulfur nucleophiles.

Fanconi Anämie ist eine seltene rezessiv vererbte Erkrankung, deren zu Grunde liegende Enzymdefekte in ein Netzwerk unterschiedlichster DNA-Reparaturproteine eingewoben sind. Phylogenetisch sind uns Vögel relativ nahe verwandt, was sie zu einem guten Modellorganismus jenseits der Säugetiermodelle macht. Eine von Hühnerzellen abgeleitete Zelllinie (DT40) wurde bereits schon breit eingesetzt um die Funktion des FA-Signalwegs zu erforschen. Nachdem auch das Hühnergenom vollständig entschlüsselt wurde, konnten zu fast allen FA-Genen Orthologe gefunden werden. Unter den zahlreichen FA-Genen sind für diese Arbeit vor allem FANCC und -G von Bedeutung, da beide Gene auf dem Z-Geschlechtschromosom des Huhns liegen und eine Inaktivierung des zweiten Z-Chromosoms beim Hahn äquivalent zur X-Inaktivierung beim Menschen nicht stattfindet. Somit sollte es ein ´natürliches´ Gendosisungleichgewicht zwischen den Geschlechtern geben. Im durchgeführten Southern Blot konnte keine geschlechtsspezifisch weibliche Bande (für FANCC und -G) gefunden werden. Somit ist davon auszugehen, dass die FA-Gene C und G ausschließlich auf dem Z-Chromosom lokalisiert sind. Dies wurde auch nochmals mittels FISH bestätigt - beide Gene fanden sich auf dem kurzen Arm des Z-Chromosoms (FANCC zentromernah, FANCG zentromerfern). Aus Studien mit DT40 Zellen ist bereits bekannt, dass FA defiziente Zellen ähnlich wie humane FA-Zellen eine Hypersensitivität gegenüber Substanzen zeigen, die DNA-crosslinks verursachen. In Anlehnung an die humane FA-Diagnostik wurden die neu etablierten embryonalen Fibroblasten mit unterschiedlichen Konzentrationen und Einwirkzeiten von MMC behandelt und die Schäden ausgewertet. In allen Untersuchungen trugen die weiblichen Zellen mehr Schäden davon als die männlichen. Bei niedrigen Konzentrationen zeigte sich dies nur als Trend, bei höheren MMC-Konzentrationen und längeren Einwirkzeiten fanden sich bei fast allen durchgeführten Untersuchungen auch statistisch signifikante Unterschiede. Somit ergibt sich aus dieser Arbeit ein deutlicher Hinweis auf ein funktionelles Ungleichgewicht zwischen Henne und Hahn was die DNA-Reparatur nach Schädigung durch MMC angeht. / Fanconi anemia is a rare recessive disorder whose underlying enzyme deficiencies are woven into a network of various DNA repair proteins. Phylogenetically related birds are relatively close to us, which makes it a good model organism beyond the mammalian models. A cell line derived from chicken cells (DT40) has already been widely used to study the FA pathway. Even after the chicken genome was completely decoded, orthologs could be found for almost all FA genes. Among the numerous FA genes are mainly FANCC and G of importance since both genes are located on the Z sex chromosome of chicken. An inactivation of the second Z chromosome as the inactivation of the x chromosom in human does not take place. Thus there should be a 'natural' imbalance of the gen dose between the sexes. Southern blot showed no gender female band (for FANCC and G). Thus it can be assumed that the FA genes C and G are exclusively localized on the Z chromosome. This was also confirmed by FISH, both genes were located on the short arm of the Z chromosome. From studies in DT40 cells is already known that FA deficient cells, similar to a human FA cell show hypersensitivity to substances that cause DNA crosslinks. Based on the human FA diagnosis, newly established embryonic fibroblasts were treated with different concentrations of MMC and chromosomal the damage was evaluated. In all studies, the female cells contributed more damage than the male. At low concentrations, this was only shown as a trend, at higher MMC concentrations and longer exposure times there was a significant differences. Thus, results from this study, a clear indication of a functional imbalance between hen and rooster in terms of DNA repair after damage by MMC.

published_or_final_version / Chemistry / Doctoral / Doctor of Philosophy

Mitomycin C (MMC), obtained from Streptomyces caespitosus, a clinically useful antitumour antibiotic, is the archetypical quinone bioreductive alkylating agent. The reductive activation mechanism of MMC, involves quinone reduction sequentially activating electrophilic sites in the drug molecule (C-l and C-lO for MMC). This research project was designed to investigate the role of the C-lO in alkylation processes by preparing compounds in which the electrophilicity at C-l is much reduced by substituting a cyclopropane for the aziridine ring. The resulting pyrrolo[I,2-a]indole, cyclopropamitosenes, could on reductive activation, by either 1- or 2-electron processes, followed by elimination of the carbamate, generate a powerful electrophile capable of alkylating DNA (or other nucleophiles) at C-lO .. A range of compounds was prepared utilising the azidocinnamate decomposition route to substituted indoles and an intramolecular [3 + 2] cycloaddition strategy was employed to synthesise the pyrrolo[I,2-a]indole nucleus. The rapid ring opening of cyclopropylcarbinyl radicals is briefly outlined. The reduction-initiated ring opening of the cyclopropane ring is investigated, thereby establishing its relevance to the potent bioreductive anticancer action of the cyclopropamitosenes, novel analogues of MMC. The design and synthesis of fused [I,2-a]indoles without the cyclopropane ring, is examined for comparative purposes. The key step in the synthesis is the formation of the [I,2-a ]indole nucleus via a radical cyclisation. Biological data were recorded for the cyclopropamitosenes and correlated with their structures.

Novel mitosenes substituted at the 6-position were synthesized for antineoplastic screening. More than 26 new compounds were made by two synthetic routes. A Nenitzescu-type synthesis provided ethyl 1-acetoxy-2,3-dihydro-5,8-dione-7-methoxy-1H-pyrrolo{1,2-a}indole-9-carboxylate. However, selective reduction of this ester could not be achieved satisfactorily. A more practical route via annelation of a commercially available indole was successful in completing the planned scheme of 6-substituted mitosene congeners. The third ring (pyrrolidine) was added by condensation of ethyl acrylate with ethyl 5-methoxyindole-2-carboxylate. After decarboxylation at position 2, the ketone at position 1 was reduced and acetylated. Then the carbon at the 9 position was introduced by Vilsmeier-Haack formylation and the quinone moiety was synthesized via a nitration, reduction, and oxidation sequence. Subsequently, the aldehyde was most satisfactorily reduced to an alcohol with sodium borohydride and the quinone was regenerated with Fremy's salt. 1-acetoxy-6-desmethyl-7-methoxymitosene was made by forming a carbamate at position 9 by treatment of the 9-alcohol with phenyl chloroformate and displacing the phenoxy group with ammonia. Other 1,6,7-substituted mitosene congeners were made using N-methylcarbamate formation via methyl isocyanate and the 9-alcohol. The 6-chloro and 6-bromo analogs were formed by treatment of the 6-H congener, 1-acetoxy-2,3-dihydro-5,8-dioxo-9-(hydroxymethyl)-7-methoxy-1H-pyrrolo{1,2-a}indole methylcarbamate, with the desired halogen in acetic acid and sodium acetate. The 7-methoxy group could be displaced by ammonia for the 6-bromo compound and by pyrrolidine for the 6-H compound to form respectively the 7-amino-6-bromo and 7-pyrrolidino-6-H 1-acetoxy-2,3-dihydro-5,8-dioxo-9-(hydroxymethyl)-1H-pyrrolo{1,2-a}indole methylcarbamates. The 6-methyl analog (1-acetoxy-7-methoxy-N-methyl-carbamoylmitosene) was made from a previously synthesized precursor. Attempted syntheses of the 6-azido and 6-amino analogs by displacing the 6-bromo substituent with sodium azide were met by gross rearrangement of the resulting adducts. Preliminary antitumor screening against P388 leukemia in mice showed these analogs to be too inactive for use as antineoplastic agents. The 6-methyl substituent was shown to be most potent in bacteriophage induction in E. coli for this series of 6-substituted mitosene analogs.

No description available.

M.Med. Faculty of Health Sciences, University of the Witwatersrand, 2009 / Purpose To compare the outcomes of primary viscocanalostomy with trabeculectomy augmented with mitomycin C (MMC) in black South African patients with primary open-angle glaucoma (POAG). Method A prospective, randomized study was conducted over a four year period. Fifteen black South African patients with bilateral open-angle glaucoma requiring surgery that met the inclusion criteria and gave informed consent to participate in the trial were randomized to receive either a viscocanalostomy or a trabeculectomy with MMC in the first eye requiring surgery. The fellow eye then received the alternate procedure. Patients were followed up for two years postoperatively. Results There were no significant differences between the two surgical groups preoperatively. Twelve eyes in each group were followed for twenty-four months. In both groups the intraocular pressure (IOP) was significantly reduced post-operatively (p < 0.01) and the average number of medications used per eye was significantly reduced (p < 0.02). At twenty-four months, complete success (IOP less than or equal to 18mmHg without glaucoma medication and with no evidence of glaucoma progression) was seen in 75% of eyes undergoing trabeculectomy with MMC that completed the follow up, but in only 33% of eyes undergoing viscocanalostomy (p = 0.0498). Survival curves for both success and qualified success (IOP less than or equal to 18mmHg with glaucoma medications) in the two surgical procedures confirmed the superiority of trabeculectomy with MMC over viscocanalostomy. Conclusion Viscocanalostomy may offer some advantages because it is less invasive, but intraocular pressure control appears to be superior with trabeculectomy with MMC and this continues to be the filtering procedure of choice for the management of glaucoma in black South African patients.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第22884号 / 医博第4678号 / 新制||医||1048(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 長船 健二, 教授 羽賀 博典, 教授 椛島 健治 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

by Chan Hung Chuen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 143-151). / Abstract also in Chinese. / Acknowledgments --- p.i / Abstract --- p.ii / Abstract (Chinese version) --- p.v / Abbreviations --- p.viii / Content --- p.ix / Chapter CHAPTER ONE --- INTRODUCTION / Chapter 1. --- Free radical and free radical-mediated antitumor drugs --- p.1 / Chapter 2. --- Mitomycin C (MC) / Chapter 2.1 --- Drug actions of MC --- p.2 / Chapter 2.2 --- Adverse side effects of MC --- p.5 / Chapter 3. --- Doxorubicin (DOX) / Chapter 3.1 --- Drug actions of DOX --- p.7 / Chapter 3.2 --- Adverse side effects of DOX --- p.8 / Chapter 4. --- Antioxidants --- p.14 / Chapter 5. --- Effects of exogenous ATP on the antitumor activity of Doxorubicin and Mitomycin C / Chapter 5.1 --- Glutathione (GSH) and related enzymes --- p.17 / Chapter 5.2 --- Glutathione (GSH) and Anticancer Quinones --- p.19 / Chapter 5.3 --- Glutathione and the cardiac toxicity of the anticancer drugs --- p.20 / Chapter 5.4 --- Glutathione depletion in tumor cells by exogenous ATP --- p.21 / Chapter 6. --- Aim of research --- p.24 / Chapter CHAPTER TWO --- THE EFFECT OF ANTIOXIDANTS ON DOXORUBICIN- OR MITOMYCIN C-INDUCED CYTOTOXICITY ON HUMAN TUMOR AND NORMAL CELL LINES / Chapter 2.1 --- Introduction --- p.26 / Chapter 2.2 --- Materials and Methods --- p.28 / Chapter 2.3 --- Results --- p.36 / Chapter 2.4 --- Discussion --- p.60 / Chapter CHAPTER THREE --- STUDY OF CARDIOPROTECTIVE EFFECTS OF ANTIOXIDANTS AGAINST DOXORUBICIN- OR MITOMYCIN C-INDUCED TOXICITY BY LANGENDORFF PERFUEED ISOLATED RAT HEART MODEL / Chapter 3.1 --- Introduction --- p.64 / Chapter 3.2 --- Materials and Methods --- p.67 / Chapter 3.3 --- Results --- p.75 / Chapter 3.4 --- Discussion --- p.76 / Chapter CHAPTER FOUR --- THE EFFECT OF ANTIOXIDANTS DURING CHEMOTHERAPY OF DOXORUBICIN OR MITOMYCIN C IN TUMOR-BEARING MICE / Chapter 4.1 --- Introduction --- p.78 / Chapter 4.2 --- Materials and Methods --- p.80 / Chapter 4.3 --- Results --- p.83 / Chapter 4.4 --- Discussion --- p.93 / Chapter CHAPTER FIVE --- HISTOLOGICAL STUDY AND LIPID PEROXIDATION STUDY OF PROTECTIVE EFFECT OF ANTIOXIDANTS IN TUMOR-BEARING MICE TREATED WITH DOXORUBICIN OR MITOMYCIN C / Chapter 5.1 --- Introduction --- p.95 / Chapter 5.2 --- Materials and Methods --- p.98 / Chapter 5.3 --- Results --- p.103 / Chapter 5.4 --- Discussion --- p.117 / Chapter CHAPTER SIX --- EFFECT OF EXOGENOUS ATP ON THE ANTITUMOR ACTIVITY OF DOXORUBICIN AND MITOMYCIN C ON CULTURED HUMAN HEPATOMA CELLS / Chapter 6.1 --- Introduction --- p.122 / Chapter 6.2 --- Materials and Methods --- p.124 / Chapter 6.3 --- Results --- p.126 / Chapter 6.4 --- Discussion --- p.136 / Chapter CHAPTER SEVEN --- CONCLUSION / Chapter 7.1 --- Conclusion --- p.139 / Chapter 7.2 --- Future perspective --- p.141 / Bibliography --- p.142