Kinobeads: A Chemical Proteomic Approach for Kinase Inhibitor Selectivity Profiling and Target Discovery

Reinecke, Maria; Heinzlmeir, S.; Wilhelm, Mathias; Médard, Guillaume; Klaeger, Susan; Küster, Bernhard

doi:10.1002/9783527818242.ch4

Cited by 13 publications

(17 citation statements)

References 82 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…There are many examples of different types of qualitative assays that have been used in drug repurposing approaches. Among the most common are immobilization or affinity chromatography, where either the target or the drug are immobilized on a matrix or column followed by exposure to a drug library or potential binding targets [57]. The complexes that have formed can then be eluted and identified using analytic methods.…”

Section: Binding Assaysmentioning

confidence: 99%

Drug Repurposing Techniques in Viral Diseases

Zhang¹,

Oerlemans²,

Wang³

et al. 2022

Drug Repurposing - Molecular Aspects and Therapeutic Applications

View full text Add to dashboard Cite

Since the advent of the twentieth century, several severe virus outbreaks have occurred—H1N1 (1918), H2N2 (1957), H3N2 (1968), H1N1 (2009) and recently COVID-19 (2019)—all of which have posed serious challenges to public health. Therefore, rapid identification of efficacious antiviral medications is of ongoing paramount importance in combating such outbreaks. Due to the long cycle of drug development, not only in the development of a “safe” medication but also in mandated and extensive (pre)clinical trials before a drug can be safely licensed for use, it is difficult to access effective and safe novel antivirals. This is of particular importance in addressing infectious disease in appropriately short period of time to limit stress to ever more interlinked societal infrastructures; including interruptions to economic activity, supply routes as well as the immediate impact on health care. Screening approved drugs or drug candidates for antiviral activity to address emergent diseases (i.e. repurposing) provides an elegant and effective strategy to circumvent this problem. As such treatments (in the main) have already received approval for their use in humans, many of their limitations and contraindications are well known, although efficacy against new diseases must be shown in appropriate laboratory trials and clinical studies. A clear in this approach in the case of antivirals is the “relative” simplicity and a high degree of conservation of the molecular mechanisms that support viral replication—which improves the chances for a functional antiviral to inhibit replication in a related viral species. However, recent experiences have shown that while repurposing has the potential to identify such cases, great care must be taken to ensure a rigourous scientific underpinning for repurposing proposals. Here, we present a brief explanation of drug repurposing and its approaches, followed by an overview of recent viral outbreaks and associated drug development. We show how drug repurposing and combination approaches have been used in viral infectious diseases, highlighting successful cases. Special emphasis has been placed on the recent COVID-19 outbreak, and its molecular mechanisms and the role repurposing can/has play(ed) in the discovery of a treatment.

show abstract

Section: Binding Assaysmentioning

confidence: 99%

Drug Repurposing Techniques in Viral Diseases

Zhang¹,

Oerlemans²,

Wang³

et al. 2022

Drug Repurposing - Molecular Aspects and Therapeutic Applications

View full text Add to dashboard Cite

show abstract

“…In the latter, the benzimidazole's nitrile substituent was replaced by a bulkier 7). 79 In a chemoproteomic kinobead selectivity profiling experiment 80…”

Section: Tbk1 and Ikkεmentioning

confidence: 99%

Chemical Probes for Understudied Kinases: Challenges and Opportunities

et al. 2021

View full text Add to dashboard Cite

Over twenty years after the approval of the first-in-class protein kinase inhibitor imatinib, the biological function of a significant fraction of the human kinome remains poorly understood while most research continues to be focused on few well-validated targets. Given the strong genetic evidence for involvement of many kinases in health and disease, the understudied fraction of the kinome holds a large and unexplored potential for future therapies. Specific chemical probes are indispensable tools to interrogate biology enabling proper preclinical validation of novel kinase targets. In this Perspective, we highlight recent case studies illustrating the development of highquality chemical probes for less-studied kinases and their application in target validation. We spotlight emerging techniques and approaches employed to the generation of chemical probes for protein kinases and beyond and discuss the associated challenges and opportunities.

show abstract

“…17 Kinobeads work via solid supports that are irreversibly attached to a library of promiscuous kinase inhibitors that bind to kinases present in cell or tissue lysates. After bead washing and proteolytic digestion, bound kinases are quantified based on the release of signature peptide fragments analyzed by LC-MS. 18 In the presence of a competitive free inhibitor, less enzyme binds to the beads, leading to a concentration-dose–dependent reduction of signatures peptides(s) associated with free inhibitor bound kinases ( Fig. 3 ).…”

Section: Multiplex Cellular Kinase Profiling Can Enable Hypotheses Of Off-target Inhibitor Effectsmentioning

confidence: 99%

“…Using this platform, at least 40 advanced kinase inhibitors (including vermurafenib) were also shown to bind to the heme-synthesizing enzyme ferrochelatase (FECH). 18 Vermurafenib ( Fig. 3 ), a potent and selective azaindole inhibitor of BRAF V600E , was discovered in a fragment-based drug design SAR campaign that used a biochemical functional assay that monitored inhibition of phosphorylation of a fluorescence resonance energy transfer (FRET)–peptide substrate via a coupled kinase/protease enzymatic assay ( Suppl.…”

Section: Multiplex Cellular Kinase Profiling Can Enable Hypotheses Of Off-target Inhibitor Effectsmentioning

confidence: 99%

“…S2 ). 27–29 Both LY2811376 and AMG-8718 were dropped from clinical testing, as these inhibitors caused an accumulation of autofluorescent material in the retinal pigment epithelium (a nonregenerating layer of cells) in rats, eventually leading to severe visual impairment. 30 BACE1 knockout mice dosed with LY2811376 and PF-9283 still showed this toxicity, eluding to an off-target mechanism.…”

Section: Proteomics Enables Investigation Of Ocular Toxicity In Alzheimer’s Diseasementioning

confidence: 99%

See 1 more Smart Citation