hCAIX/XII-IN-5 (Coumarin 9a) is an inhibitor of carbonic anhydrase (CA) that shows excellent hCA IX/XII selectivity with Kis of 93.3 and 85.7 nM over hCA I and hCA II, respectively. hCAIX/XII-IN-5 has anti-proliferative activities to cancer cells that can delay cell cycle and induce apoptosis [1].

hCAIX/XII-IN-5 (2.36-32.17 μM) treatment shows both anti-proliferative activities to MCF-7 and PANC-1 cells under the normoxic and hypoxic condition [1]. hCAIX/XII-IN-5 (2.69 μM; 24 h) treatment shows accumulation of MCF-7 cells in the Sub-G 1 phase and cell arrest in G 2 -M phase [1]. hCAIX/XII-IN-5 (2.69 μM; 24 h) treatment increases the percent of annexin V-FITC-positive apoptotic cells, including both the early and late apoptotic cells (UR+LR) [1]. Cell Proliferation Assay [1] Cell Line: MCF-7 and PANC-1 cells Concentration: 2.36-32.17 μM Incubation Time: Result: Exerted anti-proliferative activities to MCF-7 and PANC-1 cells (IC 50 =2.69 μM and 32.17 μM, respectively) under the normoxic condition, and moderate activitits against both MCF-7 and PANC-1 cells (IC 50 =16.36 μM and 11.78 μM, respectively) under the hypoxic condition. Cell Cycle Analysis [1] Cell Line: MCF-7 cells Concentration: 2.69 μM Incubation Time: 24 hours Result: Resulted in a significant increase in the percentage of cells at Sub-G 1 by 14-folds with significant increase in cell percentage in the G 1 -M phase by 2.3-folds compared to the control. Apoptosis Analysis [1] Cell Line: MCF-7 cells Concentration: 2.69 μM Incubation Time: 24 hours Result: Resulted in a significant increase in the percent of annexin V-FITC-positive apoptotic cells, including both the early (from 0.61% to 6.77%) and late apoptotic (from 0.27% to 14.84%) cells (UR+LR), about 11- and 55-fold increase, respectively, as compared with the control.