The research on anticancer drugs has always been a focus. Indole derivatives can help resist free radicals, while di-indole derivatives have become anticancer agents. Calcium/calmodulin-dependent protein kinase II (CaMKII) inhibitors are one of the important research directions for targeted cancer drugs. Inhibiting CaMKII can reduce the proliferation and survival of various cancer cells, but there are currently no CaMKII inhibitor drugs available. This study uses triindole as the main framework to develop derivatives as CaMKII inhibitors, hoping to apply them as anticancer agents. The computer software Discovery Studio 2016 was used to simulate the binding energies of different triindole derivatives molecular models with CaMKIIα (PDB: 2VZ6). Compound 3, which had a higher binding energy, was selected and extended for structure-activity relationship (SAR) optimization, leading to a series of high-efficiency drug synthesis and purification work. Through biological cell activity testing, the lead compound 3-1 showed high toxicity to cancer cells and a good inhibitory effect on CaMKIIα, meeting the needs of cancer treatment. Further optimization of this structure will be carried out, and cell signaling pathways and animal experiments will be conducted.