This project offers a strategic combinatorial therapeutic approach to target glioblastoma multiforme (GBM) through nanodrug delivery of a chemotherapeutic agent and monoclonal antibodies. The project design focuses on a pentavalent therapeutic nanovaccine formulation that can specifically target GBM using hollow gold nanoparticles (AuNPs). The main aim of this project is to develop a multipotent theragnostic nanovaccine (TNVax) for targeting malignant GBM cells. The TNVax was designed as follows: the hollow gold nanocages (AuNcgs) passivated with a extremophilic sulfated polysaccharide, Mauran (MR) that was chemically bound with 5-fluorouracil (5FU). This nano-complex was functionalised with anti-CD133 antibody and anti-PD- L1 antibody for specific targeting of CD133 antigen and inhibiting the immune suppression induced by PD-L1 expression in GBM, respectively. The proposed schematic representation of the synthesis demonstrating the step by step fabrication of nanoformulations is depicted in the attached figure.
Hollow AuNcgs coated with muran- 5-Fluorouracil complex were prepared and characterised. These nanocomplex (TNVax nanoparticles) were functionalised with anti-CD133 and anti-PD-L1 antibodies for targeting and functional properties. A detailed characterization of these nanoparticles were performed using various physical, chemical and biological methods. TNVax nanoparticles were tested for anticancer activities against GBM and specificity on targeting were tested. Apart from that the immunomodulation occurring at GBM microenvironment on TNVax treatment was also aimed. The activity of TNVax in tumour directed immune response and suppression of PD-L1 signalling was also aimed to determine. In vitro cell toxicity analysis of TNVax showed promising anticancer effect against GBM cells. However, the release of free drug, 5FU was observed less compared to its release as a small-molecule complex from the polysaccharide prodrug complex. The reason for this was well demonstrated using chemical characterisations using XPS. Targeting of GBM with TNVax was shown using microscopy and the pharmacokinetic (PK) model for TNVax release was also set-up and demonstrated. GBM tumour model was developed and growth kinetics of GL261 cells to form tumour was studied using C57BL6 mice. However, in vivo efficacy and immunostimulation needs to be completed as the formulation requires certain chemical alterations in the synthetic process. The results achieved were disseminated through conferences, seminars, workshops, social media and web pages.
