Search

Novel photoswitches with features complementary to the well-established azobenzenes are increasingly driving high-precision research in photopharmacology in cells and in vivo. Styrylthiazolium (StyTz) and styrylbenzothiazolium (StyBtz) are cellularly untested E/Z-isomerisation photoswitches which are nearly isosteric to azobenzenes, but have distinct photophysical and physicochemical properties: including ca. 60 nm red-shifted π→π* absorption, self-reporting fluorescence, Z→E relaxation that matches typical biological timescales, and good solubility due to their permanent positive charge. We here tested StyTz and StyBtz for their potential as photopharmaceutical scaffolds, by applying them to photocontrol the dynamics of the microtubule cytoskeleton. We observed light-specific disruption of microtubule network architecture and antiproliferative activity, with a structure-activity relationship matching expectations for tubulin inhibitors. However, while testing the lead StyBtz2 for its molecular mechanism of action, we found that it did not inhibit microtubule dynamics. We tracked its localisation in live cells by relying on its self-reporting fluorescence, observing accumulation of E-StyBtz2 into mitochondria; after several minutes of illumination it was then released into the cytosol concomitant with blebbing and cell death. We interpret this as light-dependent catastrophic rupturing of mitochondria on acute timescales. We conclude that StyTz/StyBtz can be interesting photopharmaceutical scaffolds for addressing mitochondrial, rather than cytosolic, targets.

Very preterm infants are at high risk for suboptimal nutrition in the first weeks of life leading to insufficient weight gain and complications arising from metabolic imbalances such as insufficient bone mineral accretion. We investigated the use of a novel set of standardized parenteral nutrition (PN; MUC PREPARE) solutions regarding improving nutritional intake, accelerating termination of parenteral feeding, and positively affecting growth in comparison to individually prescribed and compounded PN solutions. We studied the effect of MUC PREPARE on macro- and micronutrient intake, metabolism, and growth in 58 very preterm infants and compared results to a historic reference group of 58 very preterm infants matched for clinical characteristics. Infants receiving MUC PREPARE demonstrated improved macro- and micronutrient intake resulting in balanced electrolyte levels and stable metabolomic profiles. Subsequently, improved energy supply was associated with up to 1.5 weeks earlier termination of parenteral feeding, while simultaneously reaching up to 1.9 times higher weight gain at day 28 in extremely immature infants (

Background: Hemithioindigo is a promising molecular photoswitch that has only recently been applied as a photoswitchable pharmacophore for control over bioactivity in cellulo. Uniquely, in contrast to other photoswitches that have been applied to biology, the pseudosymmetric hemithioindigo scaffold has allowed the creation of both dark-active and lit-active photopharmaceuticals for the same binding site by a priori design. However, the potency of previous hemithioindigo photopharmaceuticals has not been optimal for their translation to other biological models.Results: Inspired by the structure of tubulin-inhibiting indanones, we created hemithioindigo-based indanone-like tubulin inhibitors (HITubs) and optimised their cellular potency as antimitotic photopharmaceuticals. These HITubs feature reliable and robust visible-light photoswitching and high fatigue resistance. The use of the hemithioindigo scaffold also permitted us to employ a para-hydroxyhemistilbene motif, a structural feature which is denied to most azobenzenes due to the negligibly short lifetimes of their metastable Z-isomers, which proved crucial to enhancing the potency and photoswitchability. The HITubs were ten times more potent than previously reported hemithioindigo photopharmaceutical antimitotics in a series of cell-free and cellular assays, and allowed robust photocontrol over tubulin polymerisation, microtubule (MT) network structure, cell cycle, and cell survival.Conclusions: HITubs represent a powerful addition to the growing toolbox of photopharmaceutical reagents for MT cytoskeleton research. Additionally, as the hemithioindigo scaffold allows photoswitchable bioactivity for substituent patterns inaccessible to the majority of current photopharmaceuticals, wider adoption of the hemithioindigo scaffold may significantly expand the scope of cellular and in vivo targets addressable by photopharmacology.

Druglike small molecules with photoswitchable bioactivity-photopharmaceuticals-allow biologists to perform studies with exquisitely precise and reversible, spatial and temporal control over critical biological systems inaccessible to genetic manipulation. The photoresponsive pharmacophores disclosed have been almost exclusively azobenzenes, which has limited the structural and substituent scope of photopharmacology. More detrimentally, for azobenzene reagents, it is not researchers' needs for adapted experimental tools, but rather protein binding site sterics, that typically force whether the trans (dark) or cis (lit) isomer is the more bioactive. We now present the rational design of HOTubs, the first hemithioindigo-based pharmacophores enabling photoswitchable control over endogenous biological activity in cellulo. HOTubs optically control microtubule depolymerisation and cell death in unmodified mammalian cells. Notably, we show how the asymmetry of hemithioindigos allows a priori design of either Z- or E- (dark- or lit)-toxic antimitotics, whereas the corresponding azobenzenes are exclusively lit-toxic. We thus demonstrate that hemithioindigos enable an important expansion of the substituent and design scope of photopharmacological interventions for biological systems.

Wer schon mal in einem Projekt mitgearbeitet hat, hat wahrscheinlich die Erfahrung gemacht, dass Projekte immer anders verlaufen als geplant. Die Ursachen hierfur sind vielfaltig und nur bedingt beherrschbar. Die wichtigsten Grunde, weswegen Projekte immer wieder aus dem Ruder laufen, ahneln sich. Der haufigste ist der menschliche Faktor, denn schlieslich arbeiten in einem Projekt-Team immer viele sehr unterschiedliche Menschen mit verschiedenen Vorstellungen und Zielsetzungen zusammen. Daneben findet jedes Projekt innerhalb von unternehmerischen und gesellschaftlichen Rahmenbedingungen statt, an die sich das Projekt anpassen muss. Der dritte wichtige Faktor, der dazu fuhrt, dass Vorstellung und Wirklichkeit bei Projekten meist weit auseinanderliegen, ist die haufig unterschatzte Komplexitat von Projekten. Hierzu tragen auch Projektmanagement-Methoden bei, die ihre Anwender glauben lassen, dass sie Komplexitat nicht nur begreifbar machen, sondern sie sogar reduzieren – was in Wirklichkeit aber nicht moglich ist.

Die Prinzipien aus dem letzten Kapitel konnen Sie Schritt fur Schritt im Projekt einfuhren. Wenn Sie nur einzelne Prinzipien als Modul ubernehmen, erzielen Sie bereits erste positive Effekte. Den grosten Effekt auf Ihre Projektergebnisse erreichen Sie jedoch, wenn Sie von Anfang an die Prinzipien einfuhren. Dies umfasst alle Phasen des Projekts: Ideenfindung, Planung und naturlich am Ende die Durchfuhrung und die Kontrolle der Ergebnisse.

Unsere immer schneller werdende Arbeitswelt und die Verschiedenartigkeit der Firmen erfordern, dass eine gelungene Methodik fur besseres Projektmanagement 1.) nicht zu kompliziert und 2.) schnell umsetzbar ist. Dies hort sich im ersten Moment zwar logisch an, aber eben auch ein Stuck zu einfach.

Nicht erst die Corona-Krise hat gezeigt, dass es fur Unternehmen immer wichtiger wird, Projekte auch unabhangig von personlicher Anwesenheit aller Beteiligter durchzufuhren. Seien es Unternehmen mit mehreren internationalen Standorten, die eine gemeinsame Software einfuhren, oder Unterstutzung, die uber Off- und Nearshoring ins Projekt geholt wird: Immer wieder mussen Menschen in Projekten remote – das heist von verschiedenen Standorten aus der Ferne – zusammenarbeiten. (Als Offshoring oder Nearshoring bezeichnet man die Auslagerung bestimmter Tatigkeiten ins weiter oder weniger weit entfernte Ausland. Gerade im E-Commerce greift man gerne auf Entwickler und Entwicklerinnen aus Ost-Europa zuruck, wenn man im Inland nicht genugend qualifizierte Mitarbeiter findet – was bei schnell wachsenden Start-ups keine Seltenheit ist.)

Small molecule inhibitors provide a versatile method for studies in microtubule cytoskeleton research, since tubulin is not readily amenable to functional control using genetics. However, traditional chemical inhibitors do not allow spatiotemporally precise applications on the length and time scales appropriate for selectively modulating microtubule-dependent processes. We have synthesised a panel of taxane-based light-responsive microtubule stabilisers, whose tubulin hyperpolymerisation activity can be induced by photoisomerisation to their thermodynamically metastable state. These reagents can be isomerised in live cells, optically controlling microtubule network integrity, cell cycle repartition, and cell survival, and offering biological response on the timescale of seconds and spatial precision to the level of individual cells. These azobenzene-based microtubule stabilisers offer the possibility of noninvasive, highly spatiotemporally precise modulation of the microtubule cytoskeleton in live cells, and can prove powerful reagents for studies of intracellular transport, cell motility, and neurodegeneration.Abstract Figure

We here report the discovery of isoquinoline-based biaryls as a new scaffold for colchicine domain tubulin inhibitors. Colchicine domain inhibitors comprise a structurally diverse class of compounds offering highly desirable cytotoxic and vascular disrupting bioactivities. Current research on colchicine domain inhibitors is focused on improving in vivo robustness and tolerability: properties that are inextricably linked to the scaffold structure employed. The isoquinoline-based biaryl scaffold we now report offers high-potency tubulin inhibition with excellent robustness and druglikeness, allowing solubility, in vivo tolerability and facile late-stage structural diversification through a tolerant synthetic route. We have confirmed the tubulin-binding properties and mechanism of these isoquinoline-based biaryls through a series of cellular tests and established safe in vivo dosing parameters in mice. By addressing several problems facing the current families of inhibitors, we thus expect that this new scaffold will find a range of powerful in vivo applications towards translational use in cancer therapy.

Here we present GFP-orthogonal optically controlled reagents for reliable and repetitive in cellulo modulation of microtubule dynamics and its dependent processes. Optically controlled reagents (“photopharmaceuticals”) have developed into powerful tools for high-spatiotemporal-precision control of endogenous biology, with numerous applications in neuroscience, embryology, and cytoskeleton research. However, the restricted chemical domain of photopharmaceutical scaffolds has constrained their properties and range of applications. Styrylbenzothiazoles are an as-yet unexplored scaffold for photopharmaceuticals, which we now rationally design to feature potent photocontrol, switching microtubule cytoskeleton function off and on according to illumination conditions. We show more broadly that this scaffold is exceptionally chemically and biochemically robust as well as substituent-tolerant, and offers particular advantages for intracellular biology through a range of desirable photopharmaceutical and drug-like properties not accessible to the current classes of photoswitches. We expect that these reagents will find powerful applications enabling robust, high precision, optically controlled cell biological experimentation in cytoskeleton research and beyond.