Azenosertib, synthesized by Zentalis Pharmaceuticals, was tested on various human cancer cell lines for cell viability and proliferation. Techniques included cell proliferation assays, Jess Simple Western analysis, generation of stable cell lines, siRNA knockdown, DNA fiber assays, immunofluorescence, immunohistochemistry, high-content imaging, and in vivo studies. Statistical analysis used GraphPad Prism, with clinical trials conducted under U.S. ethical guidelines.