Synthesis of substituted 4-(indol-3-yl)quinazolines, a new class of EGFR- tyrosine-kinase-inhibitors

0\. Titelblatt 1\. Inhaltsverzeichnis I 2\. Einleitung 1 3\. Chemisch-PharmakologischerTeil 23 4\. Experimenteller Teil 128 5\. Zusammenfassung 160 6\. Literaturverzeichnis 177 7\. Anhang 184Der EGF-Rezeptor spielt eine zentrale Rolle bei der Entstehung und Progression vieler epidermaler Tumore. Ein Grossteil dieser Tumore weist eine EGFR- Ueberexpression auf. Dadurch ist die Signaltransduktion innerhalb der Zelle gestoert, was zu vermehrter Proliferation, Angiogenese, Metastasierung und Invasion sowie zu verminderter Apoptosefähigkeit der Tumorzellen fuehrt. Aufgrund dessen sind der EGFR und seine Signalwege attraktive Angriffsziele für neue targetspezifische Krebstherapeutika. Bisher ist die Strukturvielfalt der EGFR-TK-Inhibitoren stark eingeschraenkt und bezieht sich auf verschieden substituierte 4-Anilinochinazoline. Dabei wird das Anilino-NH als essenziell für die Rezeptorbindung angesehen. Das Ziel dieser Arbeit war die Entwicklung und Optimierung verschieden substituierter 4-(Indol-3-yl)chinazoline als EGFR- TK-Inhibitoren. Bei diesen Verbindungen wurde anstatt des Anilinteils eine unterschiedlich substituierte Indolgrundstruktur eingebaut. Hierbei kam ein spezielles Syntheseverfahren zur Anwendung, das eine zweifelsfreie Moeglichkeit des Erhalts eines ganz bestimmten Substitutionsmusters garantiert. Die hervorragende EGFR-TK-Inhibition belegt, dass der Rezeptor an dieser Stelle flexibel sein muss. Daher ist die Wirkstoffklasse nicht nur wie bisher auf 4-Anilinochinazoline beschraenkt, sondern die 4-Anilino-Gruppe ist durch andere Strukturen ersetzbar. Dadurch werden neue Strukturvarianten für EGFR-TK-Inhibitoren moeglich. Die Verbindungen zeigen im ultravioletten Licht starke fluoreszierende Eigenschaften. Die starke Fluoreszenz und die exzellente EGFR-TK-Inhibition bieten eine zukunftsweisende Option für eine medizinisch/pharmazeutische Anwendung dieser Substanzklasse. So koennten 4-(Indol-3-yl)chinazoline als In-vitro-Diagnostika in Zellpraeparationen Wachstumsfaktor-Rezeptoren mithilfe der Fluoreszenz der gebundenen Substanz quantitativ kenntlich machen und damit im Hinblick auf eine für den Patienten abgestimmte Therapie hilfreich sein. Außerdem wurden die Substanzen einem Test auf Zytotoxizität an der MCF-7-Brustkrebszelllinie unterzogen. Teilweise wurden hierbei ganz hervorragende Resultate erzielt, die die des als Vergleich mitgefuehrten Zytostatikums Cisplatin bei weitem übertrafen. Daher werden die Verbindungen momentan an 60 weiteren Zelllinien am National Cancer Institut in den USA getestet. Durch diese neuen Erkenntnisse eroeffnet sich ein weites Feld für moderne und innovative EGFR-TK-Inhibitoren.The development of tyrosine kinase inhibitors is a modern, innovative and promising area in the medicinal research. The aim of this present thesis was the syntheses and optimization of substituted 4-(indol-3-yl)quinazolines, a new class of EGFR-tyrosine-kinase-inhibitors. The structures of the compounds represent a more fundamental change in the pharmacophore compared with the 4-anilinoquinazolines. Searching for cross-coupling processes, an interesting pathway was discovered, including hetarylations with help of Grignard compounds. In the main step prepared indolyl-magnesium compounds were converted with 4-chloroquinazolines to give 4-(indol-3-yl)quinazolines. The synthetic pathways are suitable to guarantee the desired halogen substitution pattern in the molecules. The synthesized compounds were tested for their In- vitro EGFR-tyrosine-kinase-inhibition and their cytotoxity at the MCF-7-cell line. Partly the results shows an excellent inhibition of the EGFR-tyrosine- kinase and outstanding cytotoxic qualities. Furthermore 4-(indol-3-yl)quinazolines show strong fluorescences in the ultraviolet light, comparable with quinine-sulphate. The results of the EGFR-tyrosine-kinase- inhibition, cytotoxity and fluorescence give this new substance-class a very interesting option in diagnostic fields and in treatment of EGFR-dependent tumors. The EGF-receptor appears more flexible, as assumed until now. Therefore more structure-variations in positon 4 of the quinazolines are possible to yield quite new structures of modern and innovative EGFR-tyrosine- kinase-inhibitors

Involvement of the ABC-transporter ABCC1 and the sphingosine 1-phosphate receptor subtype S1P(3) in the cytoprotection of human fibroblasts by the glucocorticoid dexamethasone

Glucocorticoids (GC) represent the most commonly used drugs for the treatment of acute and chronic inflammatory skin diseases. However, the topical long-term therapy of GC is limited by the occurrence of skin atrophy. Most interestingly, although GC inhibit proliferation of human fibroblasts, they exert a pronounced anti-apoptopic action. In the present study, we further elucidated the molecular mechanism of the GC dexamethasone (Dex) to protect human fibroblasts from programmed cell death. Dex not only significantly alters the expression of the cytosolic isoenzyme sphingosine kinase 1 but also initiated an enhanced intracellular formation of the sphingolipid sphingosine 1-phosphate (S1P). Investigations using S1P (3) ((-/-)) -fibroblasts revealed that this S1P-receptor subtype is essential for the Dex-induced cytoprotection. Moreover, we demonstrate that the ATP-binding cassette (ABC)-transporter ABCC1 is upregulated by Dex and may represent a crucial carrier to transport S1P from the cytosol to the S1P(3)-receptor subtype

CHENILLE : Coupled beHaviour undErstaNdIng of fauLts : from the Laboratory to the fiEld

International audience<p>The understanding of the coupled thermo-hydro-mechanical behaviour of fault zones is of fundamental importance for a variety of societal and economic reasons, such as the sustainable energy transition for the safe use of natural resources (energy storage, nuclear waste disposal or geothermal energy). The overall objective of this inter-disciplinary project is to create a dataset that will allow to highlight the physical processes resulting from a thermal and hydric load on an existing, identified and characterized fault zone.</p><p> </p><p>An in situ experiment will be performed at IRSN’s Tournemire Underground Research Laboratory to evaluate the hydraulic properties and mechanical behaviour of a fault zone in a shale formation due to an increase of gas or water pressure under incremental thermal loading. This fracturing field tests will be conducted using four types of boreholes drilled from the URL : (i) one injection borehole (INJ) with one chamber measuring 10 m in length; (ii) four boreholes (H1 to H4) dedicated to host steel canister electrical heaters, (iii) 5 boreholes (S1 to S5) dedicated to the geophysical monitoring of seismic and aseismic fracturing processes, (iv) two to four boreholes (M1 to M4) to record deformation and estimate fracture location, which will help assess the seismic survey. After an initial saturation phase of the chamber, successive sequences of fluid injection tests are planned. The preliminary injection tests will be done stepwise either at constant flow or at constant pressure rate in order to obtain a steady-state flow regime at normal in situ temperatures. The hydraulic conductivity and permeability of the fault zone will be then inferred. A second stage of hydraulic testing will involve the determination of the main hydraulic parameters during a stepwise increase of temperature within the volume (maximum temperature 150°C). In the meantime, the seismological responses of the injected structures, from the static deformation to the high-frequency (100-kHz) acoustic emissions will be surveyed. The evolution of temperature and deformation will be monitored thanks to fibre optic array. In addition, a controlled seismic experiment is proposed, using coupled magnetostrictive vibrators to investigate the structural environment before and after experiment.</p><p> </p><p>Moreover, to accompany the field study, a series of laboratory experiments will be conducted to understand the chemical and structural evolution occurring within fault zones during the thermal and hydraulic loading. Experiments in climatic chambers exposing the samples to the same heat treatment as that of the in situ experiment will be carried out in order to compare the mineralogical composition evolution of the samples with those taken from the field investigated zone. Finally, a rock mechanical study, from the microscopic to the centimeter scale with monitoring of the acoustic properties will be carried out. This study will include experiments from Scanning Electron Microscope with Energy Dispersive Spectroscopy (SEM-EDS) allowing the identification of the micro-scale mechanisms of deformation localization to which it is planned to add an acoustic measurement system. In order to study the evolution of mechanical behaviour as a function of scale, experiments in triaxial press, again with acoustic monitoring, are planned.</p&gt