Discovery of new carbonic anhydrase IX inhibitors as anticancer agents by toning the hydrophobic and hydrophilic rims of the active site to encounter the dual-tail approach

The hydrophobic and the hydrophilic rims in the active site of human carbonic anhydrase IX (hCA IX) which as well contains a zinc ion as part of the catalytic core, were simultaneously matched to design and synthesize potent and selective inhibitors using a dual-tail approach. Seventeen new compounds, 5a-q, were designed to have the benzenesulfonamide moiety as a zinc binding group. In addition, N- substituted hydrazone and N-phenyl fragments were chosen as the hydrophilic and hydrophobic parts, respectively to achieve favorable interactions with the corresponding halves of the active site. All syn-thesized compounds successfully suppressed the CA IX, with IC50 values in nanomolar range from 13.3 to 259 nM. Compounds, 5h, 5c, 5m, 5e, and 5k were the top-five compounds efficiently inhibited the tumor-related CA IX isoform in the low nanomolar range (K-I & nbsp;=& nbsp;13.3, 22.6, 25.8, 26.9 and 27.2 nM, respectively). The target compounds 5a-q developed remarkable selectivity toward the tumor-associated isoforms (hCA IX and XII) over the off-target isoforms (hCA I and II). Furthermore, they were assessed for their anti-proliferative activity, according to US-NCI protocol, against a panel of fifty-nine cancer cell lines. Compounds 5d, 5k and 5o were passed the criteria for activity and scheduled automatically for evaluation at five concentrations with 10-fold dilutions. Compound 5k exhibited significant in vitro anticancer activity with GI(50)-MID; 8.68 mu M compared to compounds 5d and 5o with GI(50)-MID; 25.76 mu M and 34.97 mu M respectively. The most selective compounds 5h and 5k were further screened for their in vitro cytotoxic activity against SK-MEL-5, HCC-2998 and RXF 393 cancer cell lines under hypoxic conditions. Furthermore, 5k was screened for cell cycle disturbance, apoptosis induction and intracellular reactive oxygen species (ROS) production in SK-MEL-5 cancer cells. Finally, molecular docking studies were performed to gain insights for the plausible binding interactions and affinities for selected com-pounds within hCA IX active site.& nbsp;(c) 2022 Elsevier Masson SAS. All rights reserved.

Automated summary

In this study, potent and selective inhibitors of human carbonic anhydrase IX were designed and synthesized using a dual-tail approach. The compounds exhibited remarkable inhibitory activity against tumor-related CA IX isoform and showed potential for clinical applications.