Exploring chalcone-based azo dyes as potential carbonic anhydrase-I/II inhibitors through experimental and computational studies.

Carbonic anhydrases (CAs) are zinc-containing metalloenzymes that play essential roles in physiological processes and are recognized as important therapeutic targets for disorders such as glaucoma, epilepsy, and cancer. In this study, a series of azo-linked chalcone derivatives (1-10) was designed, synthesized, and evaluated for their inhibitory activity against human carbonic anhydrase isoforms I and II (hCA I and hCA II). All compounds demonstrated strong inhibitory activity in the low nanomolar range, with several derivatives surpassing the reference drug acetazolamide in potency. Remarkably, compound 1 exhibited the highest activity, particularly against hCA II, highlighting its potential as a lead candidate. Structure-activity relationship (SAR) analysis indicated that both the azo-linked aromatic moiety and the substitution pattern on the chalcone ring play critical roles in determining activity. Molecular docking studies revealed favorable binding interactions within the active site of the enzyme, which were further validated by molecular dynamics (MD) simulations conducted over 250 ns. In addition, in silico ADMET profiling suggested that the synthesized compounds possess acceptable pharmacokinetic properties, including good oral bioavailability, membrane permeability, and low predicted toxicity, supporting their drug-likeness. Hence, these findings demonstrate that azo-chalcone hybrids constitute a promising scaffold for the development of potent carbonic anhydrase inhibitors, with compound 1 identified as a strong candidate for further optimization and preclinical investigation.