On‐Command Regulation of Kinase Activity using Photonic Stimuli

Fleming, Cassandra L.; Grötli, Morten; Andréasson, Joakim

doi:10.1002/cptc.201800253

Cited by 24 publications

(22 citation statements)

References 47 publications

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“…An oral therapy involving kinase inhibitors may entail problems such as severe side effects or the development of resistances, which could be reduced by a more focused approach involving photoresponsive kinase inhibitors [ 8 ]. However, previously reported photoswitchable small molecule kinase inhibitors ( Figure 1 ) exhibited only slight differences in the biological activity between the metastable, irradiated, and the thermodynamically stable state [ 9 ]. These examples include three azobenzene-based kinase inhibitors with a 1.6- to 11-fold difference in biological activities upon photoswitching [ 10 , 11 , 12 ] and a protein kinase C inhibitor based on a diarylethene-like maleimide showing a higher difference with a factor of 26, but no reversible switching in aqueous solutions (similar to axitinib ( 1 ), see next paragraph) [ 13 ].…”

Section: Introductionmentioning

confidence: 99%

Photoswitchable Azo- and Diazocine-Functionalized Derivatives of the VEGFR-2 Inhibitor Axitinib

Heintze

Schmidt

Rodat

et al. 2020

IJMS

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In this study, we aimed at the application of the concept of photopharmacology to the approved vascular endothelial growth factor receptor (VEGFR)-2 kinase inhibitor axitinib. In a previous study, we found out that the photoisomerization of axitinib’s stilbene-like double bond is unidirectional in aqueous solution due to a competing irreversible [2+2]-cycloaddition. Therefore, we next set out to azologize axitinib by means of incorporating azobenzenes as well as diazocine moieties as photoresponsive elements. Conceptually, diazocines (bridged azobenzenes) show favorable photoswitching properties compared to standard azobenzenes because the thermodynamically stable Z-isomer usually is bioinactive, and back isomerization from the bioactive E-isomer occurs thermally. Here, we report on the development of different sulfur–diazocines and carbon–diazocines attached to the axitinib pharmacophore that allow switching the VEGFR-2 activity reversibly. For the best sulfur–diazocine, we could verify in a VEGFR-2 kinase assay that the Z-isomer is biologically inactive (IC50 >> 10,000 nM), while significant VEGFR-2 inhibition can be observed after irradiation with blue light (405 nm), resulting in an IC50 value of 214 nM. In summary, we could successfully develop reversibly photoswitchable kinase inhibitors that exhibit more than 40-fold differences in biological activities upon irradiation. Moreover, we demonstrate the potential advantage of diazocine photoswitches over standard azobenzenes.

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Section: Introductionmentioning

confidence: 99%

Photoswitchable Azo- and Diazocine-Functionalized Derivatives of the VEGFR-2 Inhibitor Axitinib

Heintze

Schmidt

Rodat

et al. 2020

IJMS

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“…This photocleavable protecting group (PPG) and its derivatives are amongst the most applied photoresponsive moieties and their properties have been thoroughly researched [ 28 , 29 ]. The application of NVOC-protected structures has enabled photo-control over protein function [ 30 ], dimerization [ 31 ] and degradation [ 32 ], as well as enzyme activity [ 33 , 34 , 35 ] and cytotoxicity [ 36 ].…”

Section: Resultsmentioning

confidence: 99%

A Photocleavable Contrast Agent for Light-Responsive MRI

et al. 2020

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Thanks to its innocuousness and high spatiotemporal resolution, light is used in several established and emerging applications in biomedicine. Among them is the modulation of magnetic resonance imaging (MRI) contrast agents’ relaxivity with the aim to increase the sensitivity, selectivity and amount of functional information obtained from this outstanding whole-body medical imaging technique. This approach requires the development of molecular contrast agents that show high relaxivity and strongly pronounced photo-responsiveness. To this end, we report here the design and synthesis of a light-activated MRI contrast agent, together with its evaluation using UV–vis spectroscopy, Fast Field Cycling (FFC) relaxometry and relaxometric measurements on clinical MRI scanners. The high relaxivity of the reported agent changes substantially upon irradiation with light, showing a 17% decrease in relaxivity at 0.23T upon irradiation with λ = 400 nm (violet) light for 60 min. On clinical MRI scanners (1.5T and 3.0T), irradiation leads to a decrease in relaxivity of 9% and 19% after 3 and 60 min, respectively. The molecular design presents an important blueprint for the development of light-activatable MRI contrast agents.

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“…However, although some of these approaches allow for precise temporal and spatial control of the activity of a given kinase (opto-based tools), most of them fail to regulate a single protein target, likely hiding the effect of a unique kinase-substrate by pleiotropic effects. In addition, optical manipulation is only possible up to a limited tissue depth precluding the use of opto-based in non-light permeable tissues and in many cases require the use of UV light (the vast majority of photolabile caging moieties introduced into kinases/small molecule kinase inhibitors), hindering its extensive use in vivo due to the toxicity and limited tissue penetration of UV light 9,12,13 . Photoswitchable kinases (generated by inclusion of an allosteric photosensory domain or a photodissociable dimeric domain into the kinase of interest) employ longer wavelengths (400–500 nm) for isomerization compared to the UV light used for the uncaging reactions; however, to date, only a limited alteration of kinase activity between the active and inactive form has been achieved, limiting efficient control over kinase activity.…”

Section: Introductionmentioning

confidence: 99%

In vivoregulation of fluorescent fusion proteins by engineered kinases

Lepeta

Roubinet

Kanca³

et al. 2021

Preprint

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Reversible protein phosphorylation by kinases in extensively used to control a plethora of processes essential for proper development and homeostasis of multicellular organisms. One main obstacle in studying the role of a defined kinase-substrate interaction is that kinases form complex signaling networks and most often phosphorylate multiple substrates involved in various cellular processes. In recent years, several new approaches have been developed to control the activity of a given kinase. However, most of them fail to regulate a single protein target, likely hiding the effect of a unique kinase-substrate by pleiotropic effects. To overcome this limitation, we have created protein binder-based engineered kinases for direct, robust and tissue-specific phosphorylation of target fluorescent protein fusions in vivo. We show that synthetic Rok kinases, based on the Drosophila ortholog of Rho-associated protein kinase (ROCK), are functional enzymes and can activate myosin II through phosphorylation of Sqh::GFP or Sqh::mCherry in different morphogenetic processes in a developing fly embryo. We next use the system to study the impact of actomyosin activation specifically in the developing tracheal branches and showed that ectopic activation of actomyosin with engineered Rok kinase did not prevent cell intercalation nor the formation of autocellular junctions. We assume that this approach can be adapted to other kinases and targets in various eukaryotic genetic systems.

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On‐Command Regulation of Kinase Activity using Photonic Stimuli

Cited by 24 publications

References 47 publications

Photoswitchable Azo- and Diazocine-Functionalized Derivatives of the VEGFR-2 Inhibitor Axitinib

Photoswitchable Azo- and Diazocine-Functionalized Derivatives of the VEGFR-2 Inhibitor Axitinib

A Photocleavable Contrast Agent for Light-Responsive MRI

In vivoregulation of fluorescent fusion proteins by engineered kinases

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