Insights into real-time chemical processes in a calcium sensor protein-directed dynamic library

Area of Science:

• Biochemistry
• Medicinal Chemistry
• Chemical Biology

Background:

• Dynamic combinatorial chemistry (DCC) accelerates drug discovery but often requires non-physiological conditions.
• Limited tools exist for DCC that operate effectively under physiological parameters.

Purpose of the Study:

• To develop a catalyzed protein-directed DCC system operating at low temperatures and physiological pH.
• To identify novel ligands for the calcium sensor protein NCS-1 in situ.
• To elucidate the mechanisms of molecular recognition and DCC processes.

Main Methods:

• Catalyzed protein-directed dynamic combinatorial chemistry (DCC) at low temperatures.
• Ultrafast Nuclear Magnetic Resonance (NMR) for real-time reaction intermediate analysis.
• X-ray crystallography and computational methods for structural and mechanistic elucidation.

Main Results:

• The system successfully identified ligands for the calcium sensor NCS-1 under physiological conditions.
• Ultrafast NMR provided real-time insights into acylhydrazone exchange and molecular assembly.
• A compound stabilizing the NCS-1/Ric8a complex was identified.

Conclusions:

• Protein-directed DCC under physiological conditions is a viable strategy for drug discovery.
• The identified compound shows therapeutic potential in a Drosophila model of synaptic disease.
• This work offers a new platform for in situ ligand discovery and mechanistic studies.