We have identified a small library of novel substituted 9-aminoacridine derivatives that inhibit cell proliferation of pancreatic cancer cell lines by inducing apoptosis [Goodell, J.R. et al., 2008. J. Med. Chem. 51, 179-182.]. To further investigate their antiproliferative activities, we have assessed the antiproliferative activity of these acridine-based compounds against several pancreatic cancer cell lines. All four compounds used in this study inhibited the proliferation of pancreatic cancer cell lines in vitro. In addition, we have employed a xenograft tumor model and found that these compounds also inhibit the proliferation of pancreatic cancer in vivo. In light of the potential importance of the anticancer activity of these acridine-based compounds, we have conducted a series of biochemical assays to determine the effect of these compounds on human topoisomerase II. Unlike amsacrine, these compounds do not poison topoisomerase II. Similar to amsacrine, however, these compounds intercalate into DNA in a way that they would alter the apparent topology of the DNA substrate. Thus, inhibition of the relaxation activity of topoisomerase II by these compounds has been reexamined using a DNA strand passage assay. We have found that these compounds, indeed, inhibit the catalytic activity of topoisomerase II. Thus, these novel acridine-based compounds with anti-pancreatic cancer activity are catalytic inhibitors, not poisons, of human topoisomerase II.

京都大学 / 0048 / 新制・論文博士 / 博士(医学) / 乙第13109号 / 論医博第2127号 / 新制||医||1022(附属図書館) / (主査)教授 今中 雄一, 教授 川上 浩司, 教授 髙折 晃史 / 学位規則第4条第2項該当 / Doctor of Medical Science / Kyoto University / DFAM

The problem of finding targeted medicine is a central problem in chemotherapy. From this point of view the ubiquitin-proteasome system is a highly promising object in the pharmaceutical approach. Proteasome plays a critical role in cellular protein degradation, cell cycle and apoptosis regulation. Proteasome inhibitors are substances blocking the actions of proteasome. Cancer cells are more sensitive to inhibition of the ubiquitin-proteasome system than normal cells. Therefore proteasome inhibitors have the potential to be successfully used in the cancer treatment. The study aimed to test various substances to identify possible proteasome inhibitors with the IncuCyteTM FLR image system and fluorometric microculture cytotoxicity assay. Using the IncuCyte FLR method allows for detecting changes in the molecular processes of living cells. To make proteasome inhibition visible the model cell line MelJuSoUbG76V-YFP is used which helps to detect alterations in proteasome activity by means of the yellow fluorescent protein enrichment in cells as a response to proteasome inhibition. Fluorometric microculture cytotoxicity assay is a method for the determination of cytotoxicity in human tumor cells. The study showed that substance #25 possessed a proteasome inhibitory capacity in a dose-dependent manner as demonstrated with the IncuCyte FLR image system. According to the fluorometric microculture cytotoxicity assay, substance #1 was the most stable and toxic. Substances #2 and #185 had selective toxicity against cancer cells and lower effects against normal cells. Combining IncuCyte FLR and fluorometric microculture cytotoxicity assay allows finding substances which act as proteasome inhibitors with high toxic effect.

En nanomédecine, une nouvelle approche consiste à développer des vecteurs synthétiques pour co-délivrer au sein d’une cellule tumorale, un anticancéreux ainsi qu’un siARN, capable de supprimer l’expression d’une protéine impliquée dans les mécanismes de résistance. Les travaux décrits dans ce manuscrit ont été consacrés à la synthèse de nano-vecteurs micellaires pour la délivrance simultanée de ces deux agents thérapeutiques. Une première partie décrit la synthèse et la formulation de micelles nanométriques diacétyléniques photopolymérisables conçues pour délivrer efficacement un siARN. Les propriétés d’encapsulation et de délivrance de ces micelles ont ensuite été étudiées in vitro et in vivo pour une application en thérapie combinatoire. Enfin, une dernière partie présente la fonctionnalisation par interaction électrostatique de ces vecteurs cationiques avec des anticorps préalablement modifiés par des oligonucléotides anioniques pour réaliser un ciblage actif des cellules tumorales. / In the nanomedecine field, a new approach consists in developing synthetic vectors able to co-deliver into a cancer cell, an antitumoral drug and siRNAs that target protein(s) involved in MDR. The work described in this manuscript was dedicated to the development of micellar nanovectors for the intracellular co-delivery of these two therapeutic agents. The first part details the synthesis and the formulation of nanometric photopolymerized diacetylenic micelles adapted for the delivery and intracellular release of the siRNA. Then, the encapsulation and delivery properties of these micelles, bearing histidine polar heads have been investigated in vitro and in vivo for the application of combination therapy. Finally, the last part presents the functionalization by electrostatic interaction of these cationic vectors with antibodies, priorly modified by anionic oligonucleotides. This original and versatile system allowed achieving an active targeting of tumoral cells.

A simple method is reported to synthesize an anticancer drug, NCX 4040, conveniently in mild conditions using silicon chemistry. A starting material, 4-hydroxybenzyl alcohol, was silylated selectively first to give t-butyldimethylsilyl 4-hydroxybenzyl ether, which was then converted to NCX 4040 by esterification, desilylation, hydrochlorination and nitration.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第21256号 / 医博第4374号 / 新制||医||1029(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 中山 健夫, 教授 小川 修, 教授 川村 孝 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

No description available.

no / Aspirin, formally known as acetylsalicylic (ASA), is most widely used and cheapest over-the counter drugs. It is used not only for the common fevers, headaches and inflammation, but also for reducing the risk of heart attacks. In recent years, it is also linked to anti-cancer potential. Recently the US Preventive Services Working Group (UPSTF) release aspirin as a guide for cardiovascular disease and primary prevention of colorectal cancer. Platelets have been shown to play a crucial role in cancer metastasis for many years and are proposed to have an intimate reciprocal crosstalk with cancer cells. They may alter the properties of each other and have reciprocal effects. But the exact role of platelets in modifying the tumor cell properties has not been established. In clinical, cancer patients may receive platelets from outside to treat thrombocytopenia and bleeding induced by intensive chemotherapy. Therefore understanding the exact role of platelets in carcinogenesis always is a research interest, especially when evaluating anti-cancer drugs. In this study we exam the effect of platelets on viability, proliferation and adhesion of lung cancer cells A549 in culture conditions, using different concentrations of platelet rich plasma (PRP) with and without the presence of antiplatelet drug aspirin. The tumor cell EMT transformation was also investigated under different combination of PRP and aspirin in vitro. Our data showed that low-dose of aspirin can promote cell proliferation and high-dose of aspirin could inhibit cell proliferation. High concentrations of platelet-rich plasma can inhibit cell proliferation but low concentrations of platelet-rich plasma had no significant effect on cell proliferation. Platelet-rich plasma can gather around the cell to form a gelatinous film, and this lead us to a promoted tumor cell distant metastasis model. We further found out that the combination of aspirin and PRP could increase cell viability compared to single use of PRP and Aspirin can affect cell proliferation by inhibiting platelet effects. Platelet-rich plasma reduces the adhesion of A549 cell can be attenuated by aspirin. Further works will focus on combination of different doses of aspirin and PRP to confirm the above results. Other format of aspirin (nano-form) and other NSAID inflammatory drugs like Ibuprofen will also be tested. / Abstract of conference paper.

The thesis entitled “Syntheses and Characterization of Novel Materials for Efficacious Anticancer Drug Delivery and Selective Sensing of Bioanalytes” encompasses the syntheses and characterization of various novel materials those are primarily used for efficacious pH-targeted chemotherapy, selective sensing and quantification of ATP inside a single living cell and also for specific sensing of female sex pheromone of certain agriculturally important pests. In recent era of cancer research, pH guided anticancer drug delivery is an emerging field by which not only the drug-sensitive, but also the drug-resistant cancer cell lines can be targeted efficiently. Scientists have paid lot of attentions to this area of research to design biocompatible, pH-responsive drug delivery vehicles, where most of the literatures are end up with complex, elaborated synthetic procedures and use of expensive chemicals. There are only a few reports in the literature on small molecule based drug delivery vehicles, which is not well explored. Herein some of the biocompatible, pH-sensitive lipid and short peptide sequences are synthesized in easy and short synthetic procedures and successfully tested for their efficacious anticancer drug delivery properties by various biophysical and biological techniques. A pH and reduction dual bio-responsive short peptide sequences are also generated in simple steps for the same cause. The formation of different nanostructures from the self-assembly of these short peptides is probed from high level of theoretical calculations and ultimately a well known chemotherapeutic drug, doxorubicin, has been delivered efficiently both to the drug-sensitive and drug-resistant cancer cell lines. In a particular case, in vivo study has also been performed to establish the drug delivery efficacy of those serum-stable vehicles that led to proficient reduction of tumour volume as compared to the free drug. On the other hand, a few of the molecules are synthesized and characterized by various analytical means for the selective sensing and quantification of adenosine 5’-triphosphate (ATP) inside a single living cell. Unique surface functionalized templates are also fabricated over MEMS devices for specific sensing of female sex pheromone of Helicoverpa armigera and Bactocera oleae pest in an agricultural field to detect the early pest infestation. Toward this end, an extensive study on the design, syntheses and characterization of different novel materials is presented below.

<p>During the last decades, chemotherapy with cytotoxic drugs has played a significant role in cancer therapy. It’s important to develop new anticancer drugs, and drug sensitivity testing in vitro can be used to find the right diagnosis for the newly developed substances.</p><p>The aim of this study was to investigate the cytotoxic activity of the new signal transduction modulators bortezomib, gefitinib and PKC412. The well-established substances cisplatin, cytarabine, doxorubicin and vincristin were investigated for comparison.</p><p>The activity of the cytotoxic drugs was analysed in human tumor samples from patients with ovarian carcinoma (n=16) and lymphoma (n=15) by using the Fluorometric Microculture Cytotoxicity Assay (FMCA). The testing of cellular drug resistance by FMCA was accomplished successfully in 33 out of the 34 samples (97%).</p><p>The results of this study indicated that the activity of cytotoxic drugs in tumor cells obtained from patients with ovarian carcinoma and lymphoma may be detected by the FMCA. It also suggested that bortezomib and gefitinib could represent promising agents for treatment of ovarian carcinoma and that PKC412 might be of less use for patients with this diagnose.</p>