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Structure-Activity Relationship (SAR) Study of Spautin-1 to Entail the Discovery of Novel NEK4 Inhibitors
Mathias Elsocht1, Philippe Giron2,3, Laila Maes4,5
1Research Group of Organic Chemistry, Faculty of Sciences and Bioengineering Sciences, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium.
Abstract:
Lung cancer is one of the most frequently diagnosed cancers accounting for the highest number of cancer-related deaths in the world. Despite significant progress including targeted therapies and immunotherapy, the treatment of advanced lung cancer remains challenging. Targeted therapies are highly efficacious at prolonging life, but not curative. In prior work we have identified Ubiquitin Specific Protease 13 (USP13) as a potential target to significantly enhance the efficacy of mutant EGFR inhibition. The current study aimed to develop lead molecules for the treatment of epidermal growth factor receptor (EGFR)-mutant non-small cell lung cancer (NSCLC) by developing potent USP13 inhibitors initially starting from Spautin-1, the only available USP13 inhibitor. A SAR study was performed which revealed that increasing the chain length between the secondary amine and phenyl group and introducing a halogen capable of inducing a halogen bond at position 4' of the phenyl group, dramatically increased the activity. However, we could not confirm the binding between Spautin-1 (or its analogues) and USP13 using isothermal titration calorimetry (ITC) or thermal shift assay (TSA) but do not exclude binding under physiological conditions. Nevertheless, we found that the anti-proliferative activity displayed by Spautin-1 towards EGFR-mutant NSCLC cells in vitro was at least partially associated with kinase inhibition. In this work, we present N-[2-(substituted-phenyl)ethyl]-6-fluoro-4-quinazolinamines as promising lead compounds for the treatment of NSCLC. These analogues are significantly more effective towards EGFR-mutant NSCLC cells than Spautin-1 and act as potent never in mitosis A related kinase 4 (NEK4) inhibitors (IC50~1 µM) with moderate selectivity over other kinases.
Insights
Researchers developed novel compounds targeting Ubiquitin Specific Protease 13 (USP13) to treat advanced non-small cell lung cancer (NSCLC). These new molecules show improved efficacy against EGFR-mutant NSCLC, offering a promising avenue for more effective cancer therapies.
Area of Science:
- Oncology
- Medicinal Chemistry
- Molecular Biology
Background:
- Lung cancer remains a leading cause of cancer deaths globally, with advanced stages presenting significant treatment challenges.
- Targeted therapies and immunotherapies have improved outcomes but are not curative for advanced non-small cell lung cancer (NSCLC).
- Ubiquitin Specific Protease 13 (USP13) has been identified as a potential target to enhance the efficacy of epidermal growth factor receptor (EGFR) inhibition in NSCLC.
Purpose of the Study:
- To develop novel, potent inhibitors of USP13 as lead compounds for treating EGFR-mutant NSCLC.
- To optimize initial lead molecules derived from Spautin-1 through structure-activity relationship (SAR) studies.
- To evaluate the anti-proliferative activity and kinase inhibitory profile of the developed compounds against NSCLC cells.
Main Methods:
- Structure-activity relationship (SAR) studies were conducted on Spautin-1 analogues to identify key structural modifications for enhanced activity.
- Synthesis of novel compounds, specifically N-[2-(substituted-phenyl)ethyl]-6-fluoro-4-quinazolinamines.
- In vitro evaluation of anti-proliferative activity against EGFR-mutant NSCLC cells.
- Kinase inhibition assays were performed to determine the inhibitory potency and selectivity against various kinases, including NEK4.
Main Results:
- SAR studies indicated that increasing chain length and introducing a halogen at the 4' position of the phenyl group significantly enhanced activity.
- Novel N-[2-(substituted-phenyl)ethyl]-6-fluoro-4-quinazolinamines were identified as promising lead compounds.
- These analogues demonstrated significantly greater efficacy against EGFR-mutant NSCLC cells compared to Spautin-1.
- The compounds were found to be potent inhibitors of Never in Mitosis A related kinase 4 (NEK4) with IC50 values around 1 µM, exhibiting moderate selectivity over other kinases.
Conclusions:
- N-[2-(substituted-phenyl)ethyl]-6-fluoro-4-quinazolinamines represent a promising class of compounds for the treatment of EGFR-mutant NSCLC.
- These novel inhibitors offer enhanced anti-proliferative effects compared to the parent compound Spautin-1.
- The compounds' potent NEK4 inhibition suggests a potential mechanism of action contributing to their anti-cancer activity in NSCLC.
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