Ulm, Univ., Diss., 2008.

Zugl.: Clausthal, Techn. Univ., Diss., 2008

Regensburg, Univ., Diss., 2009.

The work in this thesis was aimed to target 3 projects in which dimeric, hybrid and monomeric ligands were synthesized to target melatonin, estrogen and GPR50 receptors, respectively. The first project was the attempt of synthesis of melatonin-tamoxifen drug conjugates. Recently, a series of melatonin-tamoxifen hybrid ligands with a linker attached to the amide group of melatonin was reported as pharmacologically promising agents in the prevention and treatment of breast cancer. To examine how other attachment points, such as the ether oxygen in position 5 or substitution at C2 affect the pharmacological actions at both targets, a series of novel melatonin-tamoxifen hybrids shown below has been prepared and pharmacologically evaluated at estrogen receptor alpha and melatonin MT1 and MT2 receptors. Future screening for inhibition of viability and migration of breast cancer cells will show whether the tamoxifen drug conjugates are superior to the parent drugs and combinations thereof in their anticancer actions. Concerning the second project, melatonin MT1 and MT2 receptors are known to form homomeric and heteromeric dimers with distinct pharmacological properties. Although high-resolution structures of MT1 and MT2 receptors are available, the structures of the corresponding dimeric receptors have not been reported. To facilitate the formation of receptor dimers, and thus possibly enable the experimental determination of their structures, a series of dimeric agomelatines analogs aiming to stabilize dimeric melatonin receptors by simultaneous binding to both protomers have been developed. Future BRET and crystallographic studies on MT1 and MT2 ligand-receptor complexes are in progress. In context of GPR50 project, although currently, beside the proven inability for melatonin binding and no other ligands have been identified yet, the first potent agonists of the orphan GPR50 receptor have been identified in a functional cAMP assay. Among these analogues, the 5-methoxy-2-methyl indole which displayed a high agonistic activity at GPR50 receptor (EC50 = 0.5 nM). In this regards, various indole analogues derived from/similar to 5- methoxy-2-methylindole were synthesized in order to extend SARs. Future radioligand binding studies will allow determining the binding affinities of the ligands at GPR50 and a potential anti-cancer effect especially in breast cancer. / Die Arbeit in dieser Dissertation zielte auf drei Projekte ab, bei denen dimere, hybride und monomere Liganden synthetisiert wurden, um Melatonin-, Östrogen- bzw. GPR50-Rezeptoren zu beeinflussen. Das erste Projekt war der Versuch der Synthese von Melatonin-Tamoxifen-Wirkstoffkonjugaten. Kürzlich wurde über eine Reihe von Melatonin-Tamoxifen-Hybridliganden mit einem an die Amidgruppe von Melatonin gebundenen Linker als pharmakologisch vielversprechende Wirkstoffe für die Prävention und Behandlung von Brustkrebs berichtet. Um zu untersuchen, wie sich andere Bindungspunkte wie der Ethersauerstoff in Position 5 oder die Substitution an C2 auf die pharmakologischen Wirkungen an beiden Targets auswirken, wurde eine Reihe neuartiger Melatonin-Tamoxifen-Hybride hergestellt und pharmakologisch an Östrogenrezeptor alpha und Melatonin MT1-und MT2-Rezeptoren untersucht. Künftige Untersuchungen zur Hemmung der Lebensfähigkeit und Migration von Brustkrebszellen werden zeigen, ob die Tamoxifen-Wirkstoffkonjugate in ihrer krebshemmenden Wirkung den Ausgangsstoffen und deren Kombinationen überlegen sind. Was das zweite Projekt anbelangt, so ist bekannt, dass die Melatoninrezeptoren MT1 und MT2 homomere und heteromere Dimere mit unterschiedlichen pharmakologischen Eigenschaften bilden. Obwohl hochauflösende Strukturen von MT1- und MT2-Rezeptoren verfügbar sind, wurden die Strukturen der entsprechenden dimeren Rezeptoren noch nicht veröffentlicht. Um die Bildung von Rezeptordimeren zu erleichtern und damit möglicherweise die experimentelle Bestimmung ihrer Strukturen zu ermöglichen, wurde eine Reihe von dimeren Agomelatin-Analoga entwickelt, die darauf abzielen, dimere Melatoninrezeptoren durch gleichzeitige Bindung an beide Protomere zu stabilisieren. Zukünftige BRET- und kristallographische Studien zu MT1- und MT2-Liganden-Rezeptor-Komplexen sind in Arbeit. Im Rahmen des GPR50-Projekts wurden die ersten potenten Agonisten des verwaisten GPR50-Rezeptors in einem funktionellen cAMP-Assay identifiziert, obwohl derzeit keine Melatonin-Bindung nachgewiesen werden konnte und noch keine anderen Liganden identifiziert wurden. Zu diesen Analoga gehört das 5-Methoxy-2-Methyl-Indol, das eine hohe agonistische Aktivität am GPR50-Rezeptor (EC50 = 0,5 nM) aufweist. In diesem Zusammenhang wurden verschiedene Indol-Analoga synthetisiert, die von 5-Methoxy-2-Methylindol abgeleitet bzw. diesem ähnlich sind, um die SARs zu erweitern. Künftige Radioliganden-Bindungsstudien werden es ermöglichen, die Bindungsaffinitäten der Liganden am GPR50 und eine mögliche krebshemmende Wirkung insbesondere bei Brustkrebs zu bestimmen.

The inhibitory glycine receptors are one of the major mediators of rapid synaptic inhibition in the mammalian brainstem, spinal cord and higher brain centres. They are ligand-gated ion channels that are mainly involved in the regulation of motor functions. Dysfunction of the receptor is associated with motor disorders such as hypereklepxia or some forms of spasticity. GlyR is composed of two glycosylated integral membrane proteins α and β and a peripheral membrane protein of gephyrin. Moreover, there are four known isoforms of the α-subunit (α1-4) of GlyR while there is a single β-subunit. Glycine receptors can be homomeric including α subunits only or heteromeric containing both α and β subunits. To date, strychnine is the ligand that has the highest affinity as glycine receptor ligand. It acts as a competitive antagonist of glycine that results in the inhibition of Cl- ions permeation and consequently reducing GlyR-mediated inhibition. For a long time, the details of the molecular mechanism of GlyRs inactivation by strychnine were insufficient due to the lack of high-resolution structures of the receptor. Only homology models based on structures of other cys-loop receptors have been available. Recently, 3.0 Å X-ray structure of the human glycine receptor- α3 homopentamer in complex with strychnine, as well as electro cryo-microscopy structures of the zebra fish α1 GlyR in complex with strychnine and glycine were published. Such information provided detailed insight into the molecular recognition of agonists and antagonists and mechanisms of GlyR activation and inactivation. Very recently, a series of dimeric strychnine analogs obtained by diamide formation of two molecules of 2-aminostrychnine with diacids of different chain length was pharmacologically evaluated at human α1 and α1β glycine receptors. None of the dimeric analogs was superior to strychnine. The present work focused on the extension of the structure-activity relationships of strychnine derivatives at glycine receptors All the synthesized compounds were pharmacologically evaluated at human α1 and α1β glycine receptors in a functional FLIPRTM assay and the most potent analogs were pharmacologically evaluated in a whole cell patch-clamp assay and in [3H]strychnine binding studies. It was reported that 11-(E)-isonitrosostrychnine displayed a 2-times increased binding to both α1 and α1β glycine receptors which prompted us to choose the hydroxyl group as a suitable attachment point to connect two 11-(E)-isonitrosostrychnine molecules using a spacer. In order to explore the GlyR pocket tolerance for oxime extension, a series of oxime ethers with different spacer lengths and sterical/lipophilic properties were synthesized biologically evaluated. Among all the oxime ethers, methyl, allyl and propagyl oxime ethers were the most potent antagonists displaying IC50 values similar to that of strychnine. These findings indicated that strychnine binding site at GlyRs comprises an additional small lipophilic pocket located in close proximity to C11 of strychnine and the groups best accommodated in this pocket are (E)-allyl and (E)-propagyl oxime ethers. Moreover, 11-aminostrychnine, and the corresponding propionamide were prepared and pharmacologically evaluated to examine the amide function at C11 as potential linker. A series of dimeric strychnine analogs designed by linking two strychnine molecules through amino groups in position 11 with diacids were synthesized and tested in binding studies and functional assays at human α1 and α1β glycine receptors. The synthesized bivalent ligands were designed to bind simultaneously to two α-subunits of the pentameric glycine receptors causing a possibly stronger inhibition than the monomeric strychnine. However, all the bivalent derivatives showed no significant difference in potency compared to strychnine. When comparing the reference monomeric propionamide containing ethylene spacer to the dimeric ligand containing butylene spacer, a 3-fold increase in potency was observed. Since the dimer containing (CH2)10 spacer length was found to be equipotent to strychnine, it is assumed that one molecule of strychnine binds to the receptor and the ‘additional’ strychnine molecule in the dimer probably protrudes from the orthosteric binding sites of the receptor. / Die inhibitorischen Glycin-Rezeptoren (GlyR) gehören zu den wichtigsten Mediatoren der schnellen synaptischen Hemmung im Säugetierhirnstamm, Rückenmark und in höheren Gehirnzentren. Sie sind ligandgesteuerte Ionenkanäle, die hauptsächlich an der Regulation der motorischen Funktionen beteiligt sind. Dysfunktion des Rezeptors ist assoziiert mit motorischen Störungen wie Hyperekplexie und einigen Formen von Spastizität. GlyR sind Proteinkomplexe, die aus zwei glykosylierten integralen Membranproteinen α und β und dem peripheren Membranprotein Gephyrin bestehen. Von der α-Untereinheit sind vier Isoformen bekannt (α1-4), von der β-Untereinheit nur eine. GlyR können homomer (nur α-Untereinheiten) oder heteromer (α und ß-Untereinheiten) sein. Das Alkaloid Strychnin weist eine sehr hohe Affinität zu den GlyR auf. Es wirkt als kompetitiver Antagonist von Glycin und führt nach Bindung zu einer Hemmung des Chlorid-Ionen-Einstroms und folglich zu einer Verringerung der GlyR-vermittelten Inhibition. Lange Zeit waren die genauen Details des molekularen Mechanismus der GlyR-Inaktivierung durch Strychnin aufgrund des Fehlens von hochauflösenden Röntgenstrukturen des Rezeptors nicht bekannt; es standen nur Homologie-Modelle basierend auf Strukturen anderer cys-Loop-Rezeptoren zur Verfügung. Vor kurzem wurden eine 3.0-Å-Röntgenstruktur des humanen GlyR (α3-Homopentamer) im Komplex mit Strychnin sowie eine Kryoelektronenmikroskopie-Struktur des Zebrafisches (α1-GlyR im Komplex mit Strychnin und Glycin) veröffentlicht. Dadurch erhielt man detailliertere Informationen über die molekulare Erkennung von Agonisten und Antagonisten sowie den Mechanismen der Aktivierung und Inaktivierung von GlyR. Kürzlich wurde eine Reihe von dimeren Strychnin-Analoga, bei denen jeweils zwei Moleküle 2-Aminostrychnin durch Reaktion mit Disäuren unterschiedlicher Kettenlänge zu den entsprechenden Diamiden miteinander verknüpft wurden, pharmakologisch an humanen α1- und α1β-GlyR untersucht. Keines der dimeren Analoga war Strychnin überlegen. Die vorliegende Arbeit konzentriert sich auf der Erweiterung der Struktur-Wirkungs-Beziehungen von Strychnin-Derivaten bzgl. der Aktivität an Glycin-Rezeptoren. Die strukturellen Änderungen, die an Strychnin durchgeführt wurden, sind in Abbildung 27 dargestellt. ...

Freiburg i. Br., Univ., Diss., 2007.

Chemnitz, Techn. Univ., Diplomarb., 2008.

Der Schwerpunkt dieser Arbeit lag in der Synthese und Charakterisierung von Sulfonsäure bzw. Sulfon - und Carbonsäure - funktionalisierten Polysiloxanen für die Anwendung als selbstätzendes Dentaladhäsiv. Die grundlegende Aufgabe eines Dentaladhäsivs ist es, eine starke und langzeitstabile Verbindung zwischen Zahnhartsubstanz und Kompositmaterial zu gewährleisten. Aktuell auf dem Markt erhältliche selbstätzende Adhäsivmaterialien können jedoch das enorme Anforderungspaket teilweise nur unzureichend erfüllen. Diese enthalten meist Phosphor- bzw. Phosphonsäure - funktionalisierte Monomere, deren Hauptproblem eine ungenügende Ätzwirkung auf Dentin ist. Monomere mit stärkeren Säuregruppen, wie etwa Sulfonsäuregruppen, beschränken sich momentan auf Acrylamidomethyl-propansulfonsäure, welche lediglich in zwei kommerziell erhältlichen Adhäsivsystemen in geringen Mengen zugesetzt wird. Als Folge dieses aktuellen Forschungsbedarfs befasste sich diese Arbeit mit der Synthese und Charakterisierung neuartiger Sulfonsäure - bzw. Sulfon- und Carbonsäure -funktionalisierter Polysiloxane und der Untersuchung dieser hinsichtlich ihrer Eignung als selbstätzendes Adhäsivmaterial. / The focus of this dissertation is the synthesis and characterization of sulfonic or, respectively sulfonic and carbonic acid functionalized polysiloxanes for dental self -etching adhesives. The primary aim of dental adhesives is to provide a strong and long term stable adhesion to both the composite filling and the dental hard tissue. In addition to withstand mechanical forces a good adhesive should be able to prevent microleakage along the margins, possess a low toxicity and should be easy to use. Conventional dental adhesives based on organic monomer systems are not able to fulfill the requirements sufficiently. Currently used self - etching adhesives contain mostly phosphoric and phosphonic acid functionalized monomers, whose main problem is an insufficient corrosive property on dentin. However, the application of monomers containing stronger acidic groups, for example sulfonic acid, only concerns 2-acrylamidoethylpropane sulfonic acid so far. In this work new sulfonic acid functionalized inorganic - organic polysiloxanes were synthesized and the desired compounds analyzed with regards to their polymerization behavior, their corrosive properties on dental enamel and their bond strengths.

Multifunctional monomers on the basis of acryl- and methacryl derivatives were synthesized and different protective groups were used. After polymerization the protective groups were removed by different methods. Various initiators for the NMP of the monomers were synthesized and the reaction conditions were optimized. The results showed that NMP was not a suitable method for multifunctional acryl- and methacryl derivatives to achieve well-defined homopolymers, although it was successful for control of polymerization of styrene and block copolymerization of multifunctional acryl- and methacryl derivatives with alkoxyamine terminated polystyrene. The ATRP of multifunctional acrylates and methacrylates has been successfully performed, as well as the block copolymerization of multifunctional acrylates and methacrylates. Relatively low polydispersities of the corresponding polymers (PD=1.18-1.36) and reasonably high rates of polymerization could be achieved when Me6TREN and PMDETA were used as ligands. However, the ATRP of multifunctional acrylamides and methacrylamides failed. The RAFT-polymerization of styrene, acrylamide and acrylate using BDTB as a CTA and AIBN as an initiator afforded polymers with narrow molecular weight distribution (PD=1.13-1.26). A kinetic investigation and the further synthesis of block copolymers using dithioester-terminated homopolymers as macroCTAs showed that the RAFT polymerization of acrylamide M9b proceeded in a living manner. However, BDTB does not control the reaction of methacrylic monomers, such as methacrylates and methacrylamides. The bulk phase behavior of the block copolymers were examined by means of DSC and the surface behaviors of block copolymers as thin layers were examined with AFM. Two-phase transitions in the block copolymers were observed clearly by DSC, indicative of the appearance of phase separations, which were seen in an AFM image. In conclusion, multifunctional acryl- and methacryl derivatives failed to achieve well-defined homopolymers by NMP. However, this method was successful for block copolymerization of multifunctional acryl- and methacryl derivatives with alkoxyamine terminated polystyrene. Multifunctional acrylates and methacrylates were successfully homopolymerized and block copolymerized by ATRP. Multifunctional acrylates and acrylamides were suitable for homopolymerization and block copolymerization by the RAFT process. Thus far, it is difficult to homopolymerize multifunctional methacrylamides in controlled way.

A series of 3-(p-X-phenyl) thiophene monomers (X= –H, –CH3, – OCH3, –COOC2H5, –COCH3,–NO2) was electrochemically polymerized to furnish polymer films that could be reversibly reduced and oxidized (n- and p-doped). The oxidation potentials of the monomers and formal potentials of the n- and p-doping processes of polymers were correlated with resonance and inductive effects of the substituents on the phenyl ring as well as the semiempirically calculated heats of formation of the monomer radical cations. Moreover, the oxidation potentials of the monomers were correlated with the ionization potentials of the monomers calculated using density functional theory. The reactivity for coupling reactions and the major regioselective products of the polymerization reaction of mono- and oligo-3-phenylthiophenes are inferred from the calculated lone electron spin densities of the respective radical cations. The ionization potentials, which correspond to the energies for generating radical cations during oxidative processes were estimated.