MASARYK UNIVERSITY FACULTY OF SCIENCE DEPARTMENT OF CHEMISTRY Development of Photoactivatable Compounds Ph.D. Thesis Lenka Filipová SUPERVISOR: prof. RNDr. Petr Klán, Ph.D. BRNO 2018 Bibliographic Entry Author: Mgr. Lenka Filipová Masaryk University, Faculty of Science Department of Chemistry Title of Dissertation: Development of Photoactivatable Compounds Degree Programne: Chemistry Field of Study: Organic chemistry Supervisor: prof. RNDr. Petr Klán, Ph.D. Masaryk University, Faculty of Science Department of Chemistry Academic Year: 2018/2019 Number of Pages: 129 pages + 109 pages of appendices Keywords: reverse micelles, azobenzene, strained alkynes, cyclopropenone, Cu-free click reaction, photoCORMs, heptamethine cyanine dyes, Zincke reaction Bibliografický záznam Autor: Mgr. Lenka Filipová Masarykova univerzita, Přírodovědecká fakulta Ústav chemie Název práce: Vývoj fotoaktivovatelných sloučenin Studijní program: Chemie Studijní obor: Organická chemie Školitel: prof. RNDr. Petr Klán, Ph.D. Akademický rok: 2018/2019 Počet stran: 129 stran + 109 stran příloh Klíčová slova: reverzní micely, azobenzen, napnuté alkyny, cyclopropenon, Cu-free Click reakce, photoCORMy, barviva heptamethine cyaninová, Zincke reakce Abstract My dissertation thesis was focused on the synthesis of new photoactivatable compounds and studying their photochemical properties and behavior. The results summarized in this thesis have been published or will be submitted for the publication in the near future. The introductory part of the dissertation thesis provides previously published information related to the studied projects, including properties and behavior of reverse micelles, photoisomerization of azobenzene derivatives, click chemistry of strained alkynes, followed by application of cyclopropenones for photochemical generation of strained alkynes. Finally, the role of carbon monoxide as a signaling molecule and its potential biological effects are described. Previously reported carbon monoxide releasing molecules are also shown. In the last section of this chapter, preparation and applications of heptamethine cyanine dyes are summarized In the next chapter, Results and Disccusion, results of three research projects are summarized. The first of them is focused on the development of new photoactivatable amphiphiles and their utilization in preparation of reverse micelles. Properties of photoresponsive amphiphiles and the formation reverse micelles were studied by NMR spectroscopy at both room and sub-zero temperatures. The second project was focused on photochemically initiated Cu-free click reaction, which is widely used in the conjugation of biomolecules in the absence of metals used as catalysts. A new silacyclopropenone was developed and used in a subsequent, photoinduced Cu-free click reaction with azides and tetrazines. The third project was focused on the synthesis of new heptamethine cyanine dyes. During development of new photoactivatable compounds absorbing at higher wavelengths, Zincke reaction was utilized in the synthesis of various cyanine dyes substituted on the heptamethine chain. This method allowed the installation of different functional groups and synthesis of heptamethine cyanine dyes inaccessible by known methods. In the last part of thesis, the publications and experimental data are appended. Abstrakt Má doktorská práce byla zaměřena na syntézu nových fotoaktivovatelných sloučenin a studium jejich fotochemických vlastností. Získané výsledky popsané v této práci již byly publikovány nebo jsou sepsány a připraveny k publikaci. V úvodní kapitole jsou shrnuty dříve publikované relevantní informace, týkající se studovaných projektů, zahrnujících vlastnosti a chování reverzních micel, photoisomerizace azobenzenových derivatů, click reakce a využití napjatých alkynů a cyclopropenonů pro fotochemickou tvorbu napjatých alkynů. Dále je popsána tvorba oxidu uhelnatého jako signální molekuly v biologických systémech a jeho biologické účinky. V této části se diskutují známé molekuly uvolňující oxid uhelnatý. V poslední části této kapitoly jsou shrnuty poznatky o cyaninových barvivech, způsoby jejich přípravy a jejich využití. V následující kapitole Výsledky a diskuze jsou ukázány a shrnuty výsledky tří výzkumných projektů. První z nich se věnuje vývoji nových fotoaktivovatelných amfifilů a jejich využití při tvorbě reverzních micel. Vlastnosti samotného amfifilu a z něj vytvořených reverzních micel byly studovovány především pomocí NMR spektroskopie za laboratorní a nízké teploty. Druhý projekt se věnuje click reakci probíhající za nepřítomnosti mědi, reakci, která je využívána pro spojování biomolekul v nepřítomnosti kovů používaných jako katalyzátory. Nový silacyclopropenon byl vyvinut a použit ve světlem iniciované click reakci s azidy a tetraziny jako reagenty. Třetí projekt se zabývá syntézou nových různě substituovaných heptamethinových cyaninových barviv pomocí Zinckeho reakce. Tato metoda umožňuje instalaci mnoha funkčních skupin a syntézu cyaninových barviv, která dosud nebyla dostupná známými metodami. V poslední části práce jsou přiloženy publikace a experimentální data. Acknowledgement I would like to thank prof. Petr Klán for his supervision and opportunity to work in his group. I am grateful for fruitful discussions, support, motivation and especially for his trust in my abilities. Much thanks goes to my partner Peter. I cannot express by words how grateful I am to have you, but I hope you know it. You helped me during my entire doctoral studies, even despite the large distance between us. It was not always easy, but you stayed with me every time and endured my bad mood or crying. You are my teacher, my best friend and my husband soon. I am so grateful that we can work together and share our laboratory experience, as well as our life together. We are perfect collaborators, not only in the laboratory but also in our lives, and I hope that our collaboration will last forever. I truly and deeply love you. I thank to my current colleagues, namely Estelita, Marina, Qiuyun, Andreas, Sadegh, Mišo, Tomáš and Marek for their fruitful discussions not only about chemistry and for creating an inspiring atmosphere and fun during the work. I never experienced so nice, kind and funny group of people who like each other and work together. I am grateful that all of you are my colleagues and especially my friends. I also thank my colleagues and friends which I met during my study, Tombo and Lulu. Thanks to Luboš for technical support and providing us with everything which we ever needed in laboratory. Last but not at least I thank to my family for their support during my study, especially when they could not comprehend how I could study chemistry. Without you I would never be the person which I am today. V neposlední řadě děkuji své rodině za jejich podporu během celého studia, obzvlášť když nikdy nechápali, jak mohu studovat zrovna chemii. Vím, že to se mnou často nebylo jednoduché, ale vy jste tu vždy byli pro mě, snažili se mi všemožně pomáhat a být mi oporou. Bez vás bych nikdy nebyla tou osobou, kterou jsem dnes. “Without theory to guide him the experimenter is as lost as a sailor setting out without compass or rudder.” ‒ Leonardo da Vinci “I put my heart and my soul into my work, and I lost my mind in the process.” ‒ Vincent van Gogh © Lenka Filipová, Masaryk University, Brno 2018 Table of Contents Table of Contents Table of Contents ................................................................................................................................. 8 1. Literature Part ............................................................................................................................. 11 1.1. Introduction ........................................................................................................................ 11 1.2. Amphiphiles and its Aggregates in the Solution........................................................... 11 1.2.1. Reverse Micelles (RMs) ............................................................................................. 13 1.2.2. Mechanism of Micelle Formation ............................................................................ 15 1.2.3. Determination of Critical Micelle Concentration (CMC) ..................................... 16 1.2.4. Size of Reverse Micellar Aggregates ....................................................................... 17 1.2.5. Utilization of Reverse Micelles ................................................................................. 17 1.2.6. Reverse Micelles at Subzero Temperatures ............................................................ 17 1.3. Photoswitchable Molecules .............................................................................................. 20 1.3.1. Azobenzene Photochemistry .................................................................................... 20 1.3.2. Preparation of Azobenzene and its Derivatives .................................................... 21 1.3.3. Azobenzene Derivatives ........................................................................................... 22 1.3.4. Azobenzene Moiety Incorporated into a Self-Assembly ...................................... 22 1.4. Click Chemistry .................................................................................................................. 24 1.4.1. Bioorthogonal Click Reaction (Cu-free Click Reaction) ........................................ 25 1.4.2. Strained Alkynes .......................................................................................................