Topo I/COX-2-IN-1 (1H-30) is a potent Topo I/COX-2 inhibitor with the IC50 of 0.24 μM and 4.42 μM for COX-2 and Topo I, respectively. Topo I/COX-2-IN-1 induces apoptosis and inhibits migration of cancer cells, shows anti-cancer activity.

Topo I/COX-2-IN-1 (1H-30) exhibits potent anti-tumor properties, effectively halting tumor cell proliferation and triggering apoptosis through dose-responsive enhancement of caspase-3 activity over a 24-hour exposure (0-100 μM). Moreover, at concentrations ranging from 0.04 to 0.37 μM over 48 hours, it significantly curtails cell migration, particularly at 0.37 μM, while concurrently depressing MMP-9 expression in HGC-27 and RKO cells. Furthermore, at a 10 μM concentration and 48-hour exposure, it obstructs the activation of the NF-κB signaling pathway in cancer cells. In various colon cancer cell lines (HGC-27, RKO, HT-29, SGC-7901, and CT26.WT), it demonstrated a remarkable capability to inhibit cell proliferation, presenting IC50 values that underscore its efficacy. Additionally, apoptosis analysis revealed a marked increase in caspase-3 positive cells, particularly at a 10 μM concentration, indicating its apoptotic potency. Also, it significantly induced cell cycle arrest in the G2/M phase, highlighting its diverse mechanisms of action against cancer cell growth and spread.

Topo I/COX-2-IN-1 (1H-30), administered through intraperitoneal injection at a dosage of 100 mg/kg twice daily for 14 days, may suppress tumor growth in BALB/c mice infected with CT26.WT colon cancer cells by enhancing caspase-3 expression while reducing MMP-9 and COX-2 levels, thereby promoting apoptosis. In studies involving BALB/c mice, this regimen led to a notable decrease in tumor size and weight without adversely affecting body weight or organ health. Additionally, when tested in SD rats with the same dosage but administered once, Topo I/COX-2-IN-1 (1H-30) displayed pharmacokinetic parameters including a half-life (t 1/2) of 1.56 hours, time to reach maximum concentration (T max) of 0.67 hours, maximum serum concentration (C max) of 20.19 μg/mL, area under the curve from zero to last measurable concentration (AUC 0-t) of 18.20 mg/L h, and area under the curve from zero to infinity based on observed data (AUC 0 inf_obs) of 18.60 mg/L h.