Background: Heat shock protein 70 (HSP70) overexpression has been reported in a variety of cancers. HSP70 is transported to the cell surface and released into the environment, where it is taken up by antigen-presenting cells and presented to lymphocytes for an immunological response. In previous investigations, we extensively studied HSP70's immunotherapeutic potential against cancer. Cancer is a fatal disease, and chemotherapy drugs have been shown to be effective against several types of cancer. Several attempts have been made to improve chemotherapeutic drugs target delivery while limiting adverse effects. With the aim to minimizing off-site cytotoxicity without impairing docetaxel function in cancer cells, we established the PrDC of HSP70 isolated from cancer cells with the chemotherapy drug docetaxel, to target tumor heterogeneity and, enhance target delivery
Methodology: HSP70 isolated from cancer cells was purified using size exclusion chromatography and immunoprecipitation. To conjugate docetaxel with the primary amine groups on HSP70, a cleavable ester bond with the carboxyl group was established while preserving the structural integrity of docetaxel. The purity of HSP70 was evaluate by western blotting (WB), while NMR, FTIR, and WB were used to identify the purity of docetaxel and final conjugate. Docetaxel release from the conjugate was investigated in-vitro in four distinct buffers that replicated human philological conditions. The anticancer efficacy of docetaxel, both in combination and alone, was assessed on squamous cell carcinoma cells using the MTT, ICC, cell cycle, apoptosis, multi-drug resistance, and mitochondrial mass assay.
Results: The findings show that the conjugate of docetaxel and HSP70 has a stoichiometric ratio of 30:1. In SCC cells, docetaxel's extended release (120 hours) was correlated with a 13±2-fold rise in efficacy. Additionally, the results showed that DHSP decreases multi-drug resistance and mitochondrial mass in SCC cells while halting cells in the G2 phase of the cell cycle for an extended period of time (72 hours), which is followed by apoptosis.
Conclusions: The docetaxel-HSP70 conjunction has the potential to enhance target drug delivery and drug retention in cancer cells while lowering off-site cytotoxicity. It additionally improves docetaxel effectiveness for longer periods of time at 13–15 times lower dosages.