On-DNA hit validation methodologies for ligands identified from DNA-encoded chemical libraries

Area of Science:

• Biochemistry
• Medicinal Chemistry
• Drug Discovery

Background:

• DNA-encoded chemical libraries (DECLs) are vast compound collections used in pharmaceutical research.
• Identifying drug leads requires validating hits from DECL selections through resynthesis and affinity measurements.

Purpose of the Study:

• To develop and validate novel on-DNA strategies for hit confirmation in DECL screening.
• To enable determination of equilibrium and kinetic binding constants directly on DNA-bound ligands.

Main Methods:

• Novel on-DNA validation assays inspired by enzyme-linked immunosorbent assays (ELISA).
• Application of these methods to validate ligands targeting carbonic anhydrase II and IX.
• High-throughput DNA sequencing for initial hit identification in DECL selections.

Main Results:

• Successful validation of ligand-protein interactions directly on DNA.
• Accurate determination of equilibrium and kinetic binding constants for validated ligands.
• Demonstrated utility across ligands with varying affinities for carbonic anhydrase targets.

Conclusions:

• On-DNA validation strategies streamline the hit-to-lead process in DECL-based drug discovery.
• These methods reduce the need for extensive resynthesis and off-DNA testing.
• The developed assays provide a robust platform for pharmaceutical research and development.