Updated project metadata. Nanoparticle-based formulations bypass biological barriers, allowing for longer blood circulation times, simultaneous tumor targeting, and increased drug accumulation in tumors. They have a higher therapeutic efficiency than traditional treatment due to better specificity for effective drug targeting, no solubility, and stability issues. In our previous study, we showed that Ipomoea turpethum extract-loaded polymeric nanoparticles induce cytotoxicity in breast cancer cell lines (MCF-7 and MDA MB-231) by regulating pathways involved in apoptosis and cell proliferation 9. Many researchers demonstrated the use of tumor microenvironments for prognostic, diagnostics, and therapeutic purposes. In the present study, we have analyzed the effect of NVA-IT treatment on the tumor microenvironment via secretory proteins present in the medium of the tumor cells. We have used Nano LCMS/MS to identify the differentially expressed proteins present in the culture medium of the NVA-IT treated tumor cells compared to untreated tumor cells. The differentially expressed proteins were further analyzed using bioinformatics tools.