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

Topoisomerase I:4.42 μM

Topo I/COX-2-IN-1 (1H-30) exhibits potent anti-tumor properties by halting tumor cell proliferation and inducing apoptosis through a dose-responsive increase in caspase-3 activity over 24 hours (0-100 μM). At 0.04 to 0.37 μM over 48 hours, it significantly reduces cell migration, especially at 0.37 μM, and lowers MMP-9 expression in HGC-27 and RKO cells. At 10 μM concentration and 48-hour exposure, it inhibits the NF-κB signaling pathway activation in cancer cells. In various colon cancer cell lines (HGC-27, RKO, HT-29, SGC-7901, and CT26.WT), it demonstrates significant cell proliferation inhibition, with notable IC50 values. Apoptosis analysis reveals a marked increase in caspase-3 positive cells at 10 μM, indicating its apoptotic potency. Additionally, it induces cell cycle arrest in the G2/M phase, underscoring its multifaceted action against cancer cell growth and metastasis.

Topo I/COX-2-IN-1 (1H-30), administered via intraperitoneal injection at 100 mg/kg twice daily for 14 days, can inhibit tumor growth in BALB/c mice with CT26.WT colon cancer cells by increasing caspase-3 and reducing MMP-9 and COX-2 levels, promoting apoptosis. This regimen significantly decreased tumor size and weight without affecting body weight or organ health. In SD rats, a single dose with the same dosage showed pharmacokinetic parameters: half-life (t 1/2) of 1.56 hours, time to 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.