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Updated: Jul 30, 2026
Identification of Kinase-substrate Pairs Using High Throughput Screening
Published on: August 29, 2015
A self-immolative Kdn-glycoside substrate enables high-throughput screening for inhibitors of Kdnases
Ali Nejatie1, Cameron Proceviat1, Christina Gros1
1Department of Chemistry, Simon Fraser University, 8888 University Drive, Burnaby, BC V5A 1S6, Canada.
Abstract:
Aspergillus fumigatus, a filamentous fungus, is an opportunistic pathogen and the major causative agent of the often-fatal disease, invasive aspergillosis (IA). Current treatments for IA are limited due to their high toxicity and/or the emergence of drug resistance; therefore, a need exists for the development of new therapeutics to treat IA. The Kdnase produced by A. fumigatus plays a vital role in maintaining cell wall integrity. As there are no known Kdnases in humans, developing inhibitors of Kdnase from this fungal pathogen is a promising therapeutic approach. The rapid testing of enzymatic activity in a high-throughput screen of large chemical libraries can be an efficient way to find new small molecule lead compounds. Herein we show that a Kdn glycoside with a self-immolative cleavable aglycon is a practical and efficient substrate for a high throughput assay to identify Kdnase inhibitors. We optimized the activity assay and screened over 27,000 compounds from two bioactive chemical libraries as potential inhibitors, and we compared the hit compounds' potency towards Aspergillus terreus and Trichophyton rubrum Kdnases, two other fungal Kdnases. We validated a number of hits and these small molecules are potential leads for the development of novel therapeutics to treat invasive aspergillosis.
Insights
Developing novel therapeutics for invasive aspergillosis (IA) is crucial. Researchers identified a new high-throughput assay to screen for potential Kdnase inhibitors, leading to promising small molecules for treating this fungal infection.
Area of Science:
- Mycology
- Medicinal Chemistry
- Drug Discovery
Background:
- Invasive aspergillosis (IA), caused by Aspergillus fumigatus, is a life-threatening fungal infection.
- Existing IA treatments face challenges including toxicity and drug resistance, necessitating new therapeutic strategies.
- Fungal Kdnase is essential for Aspergillus cell wall integrity and represents a potential drug target due to its absence in humans.
Purpose of the Study:
- To develop and optimize a high-throughput screening (HTS) assay for identifying Kdnase inhibitors.
- To discover novel small molecule inhibitors of fungal Kdnase, specifically from Aspergillus fumigatus.
- To evaluate the potency of identified inhibitors against other fungal Kdnases, including Aspergillus terreus and Trichophyton rubrum.
Main Methods:
- A novel Kdn glycoside substrate with a self-immolative cleavable aglycon was synthesized for HTS.
- The Kdnase activity assay was optimized for high-throughput screening.
- Over 27,000 compounds from two bioactive chemical libraries were screened for inhibitory activity against Aspergillus fumigatus Kdnase.
Main Results:
- A practical and efficient HTS assay for Kdnase inhibitors was established.
- Screening identified numerous hit compounds with potential inhibitory activity.
- Hit compounds were further evaluated for their potency against Aspergillus terreus and Trichophyton rubrum Kdnases, with several validated.
Conclusions:
- A novel Kdn glycoside substrate enables efficient HTS for fungal Kdnase inhibitors.
- The identified small molecules represent promising lead compounds for developing new treatments for invasive aspergillosis.
- This approach offers a viable strategy for discovering novel antifungal therapeutics targeting fungal-specific enzymes.
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