Multi-kinase-IN-8 is a multi-kinase inhibitor. It effectively inhibits COX-1 (IC50 = 12.6 μM), COX-2 (IC50 = 0.05 μM), and VEGFR-2 (IC50 = 0.12 nM). Additionally, it blocks tumor-associated carbonic anhydrases CA IX and CA XII with Ki values of 31.5 nM and 386.9 nM, respectively, and induces cell cycle arrest and apoptosis (apoptosis) by upregulating Caspase-9 and Bax, while downregulating Bcl-2. Multi-kinase-IN-8 also suppresses the expression of PGE2, p-VEGFR-2, MMP-9, and HIF-1α, and demonstrates growth inhibitory activity on breast, lung, and colorectal adenocarcinomas.

COX-2:0.05 μM

Multi-kinase-IN-8 (Compound 5j) inhibits the proliferation of various cancer cell lines, with IC50 values of 1.30 μM for MDA-MB-231, 9.53 μM for MCF-7, 2.07 μM for A549, and 2.28 μM for Caco-2 cells. In Caco-2 cells, Multi-kinase-IN-8 (2.07 μM, 72 h) inhibits VEGFR-2. It also induces G1 phase cell cycle arrest and apoptosis in Caco-2 cells after 24 and 48 hours. Additionally, Multi-kinase-IN-8 downregulates HIF-1α mRNA expression in MDA-MB-231 cells and reduces levels of PGE2, total VEGFR-2, and phosphorylated VEGFR-2. The compound demonstrates selective targeting potential for tumor-associated carbonic anhydrase isoforms, particularly hCA IX, through interaction with the catalytic zinc binding site. Furthermore, in Caco-2 cells, it downregulates the expression levels of apoptotic initiator Caspase-9 and mitochondrial apoptosis markers Bcl-2 and Bax.

Multi-kinase-IN-8 (Compound 5j) demonstrated antitumor activity in a MDA-MB-231 xenograft model established in male BALB/c mice when administered subcutaneously at a dose of 50 mg/kg for 5 days.