| Abstract: |
Carbonic anhydrases IX and XII (CA-IX/XII) are Tran’s membrane metalloenzymes that display overexpression in hypoxic tumor microenvironments; both have been well established as clinically relevant targetable molecules for anticancer drug discovery. Here we describe the computational design, synthesis and biological evaluation of a series of eight novel sulfonamide-based non-covalent inhibitors (CAI−01 to CAI−08) targeting with selectivity against CA-XII over the off-target isoform CA-II. Ligand-based and structure-based drug design methods together with 100 ns molecular dynamics (MD) simulation were used to assess binding stability of Auto Dock Vine 1.2 and GLIDE XP docking approaches, respectively. Swiss ADME and pk CSM-based in silicon ADMET profiling predicted good oral bioavailability (58.2–78.6%), low her cardio toxicity risk, and compliance with Lipinski's Rule of Five for six of the eight compounds. Development of the synthesized compounds was achieved via experimental validation through enzyme inhibition assays, which showed IC50 values at NM (7.1 to 21.3 for CA-IX and 9.2 to 28.6 for CA-XII) based competitive aspect obtained from selectivity indices above 20 with respect to CA-II. The lead compound CAI-06 displayed the greatest inhibition (CA-IX IC50 = 7.1 NM; CA-XII IC50 = 9.2 nM), high selectivity between CA-IX and CA-II with a ratio of activity estimated to be 44.0, and was further identified as one of the most potent inhibitors with respect to MCF-7 cell viability across all preforming compounds at 10 uM (72.4%). Pearson correlation analysis indicated that docking calculated binding free energies highly correlated with experimental IC50 values (r = 0.912, p < 0.01). Our results therefore identify the sulfonamide-triazole scaffold as a novel chemo type for pH-selective tumor -targeting CA inhibition and therefore further pre-clinical drug development. |
