<p>Sojae semen praeparatum is a traditional Chinese medicine, and its active component, <i>Glycitin</i>, has shown potential in the treatment of non-small cell lung cancer (NSCLC). The purpose of this investigation is to examine the mechanism of action of the effective components of sojae semen praeparatum in the treatment of NSCLC, with a special emphasis on <i>Glycitin</i>, and to explore the integration of nanotechnology in delivering pharmaceutical agents. Key effective components were selected through network pharmacology analysis and functional analysis, and protein–protein interaction analysis and functional enrichment were performed using transcriptomics and metabolomics data to identify the key NSCLC-related target genes and regulatory mechanisms of action of the active components of sojae semen praeparatum. <i>Glycitin</i>-loaded NPs encapsulated in tumor-associated fibroblast membranes were developed to verify their characterization and safety, and their therapeutic effects in inhibiting the malignant phenotype of NSCLC cells through targeting the DNA topoisomerase II alpha (TOP2A) protein were validated. The results indicate that <i>Glycitin</i> exhibits significant anti-tumor activity by affecting the function of the TOP2A protein, thereby inhibiting tumor proliferation and metastasis. This research presents proof of the crucial function of <i>Glycitin</i> in managing NSCLC using sojae semen praeparatum, and sheds light on the possibilities of nanotechnology in drug delivery mechanisms, offering a novel avenue for NSCLC therapy research.</p>