Characterization of Specific N-α-Acetyltransferase 50 (Naa50) Inhibitors Identified Using a DNA Encoded Library

Area of Science:

• Biochemistry
• Structural Biology
• Medicinal Chemistry

Background:

• The N-alpha-acetylation pathway plays crucial roles in various cellular processes.
• Naa50 is a key enzyme in this pathway, and its inhibitors are sought for therapeutic applications.
• Understanding enzyme-inhibitor interactions is vital for drug development.

Purpose of the Study:

• To identify and characterize novel inhibitors of Naa50.
• To elucidate the mechanism of action and binding modes of these inhibitors.
• To assess the selectivity and cellular activity of the identified compounds.

Main Methods:

• DNA-encoded technology screening for compound identification.
• Biophysical and biochemical assays to determine binding affinities and kinetics.
• X-ray crystallography to obtain co-crystal structures of inhibitors bound to Naa50.
• Cellular target engagement assays to confirm in vivo activity and selectivity.

Main Results:

• Two novel Naa50 binders/inhibitors, compounds 3a and 4a, were identified.
• Co-crystal structures revealed detailed binding interactions, explaining affinity differences.
• Compound 4a demonstrated confirmed Naa50 engagement and selectivity over related enzymes Naa10 and Naa60.
• Structure-activity relationship studies were initiated based on analog evaluation.

Conclusions:

• Novel Naa50 inhibitors have been successfully developed using DNA-encoded technology.
• Structural and cellular data provide a strong foundation for further optimization of Naa50 inhibitors.
• The identified compounds represent promising leads for targeting the N-alpha-acetylation pathway.