Preparation, Characterization, and Biodistribution of Glutathione PEGylated Nanoliposomal Doxorubicin for Brain Drug Delivery with A Post-insertion Approach

Brain cancer treatments have been largely unsuccessful due to the blood-brain barrier. Several publications support the presence of glutathione (GSH) receptors on the surface of the BBB and consequently the products such as the 2B3-101, which is almost 5% pre-inserted GSH PEGylated liposomal doxorubicin, is under process in clinical studies. Here we conducted the PEGylated nanoliposomal doxorubicin particles that are covalently attached to the glutathione using the post-insertion technique. The post-insertion methodology is noticeably simpler, faster, and more cost-effective compared to the pre-insertion method which makes it desirable for large-scale pharmaceutical manufacturing. The 25, 50, 100, 200, and 400 ligands of the DSPE PEG(2000) Maleimide-GSH complexes were incorporated into the available Caelyx. According to the animal studies such as biodistribution, fluorescent microscopy, and pharmacokinetic studies, the 200L and 400L treatment arms were the most promising formulations compared to the Caelyx. They proved that post-inserted nanocarriers with 40 times lower levels of GSH micelles compared to the 2B3-101 have significantly increased the penetrance through the blood-brain barrier. Other tissue analysis showed that the doxorubicin will likely accumulate in the liver, spleen, heart, and lung in comparison with the Caelyx due to the expressed GSH receptors on tissues as an endogenous antioxidant. In conclusion, as was expected, the post-insertion technique was found a successful approach with more pharmaceutical aspects for large-scale production. Moreover, it is highly recommended further investigations to determine the efficacy of 5% post-inserted GSH targeted nanoliposomes versus the 2B3-101 as a similar formulation with a different preparation method.