Enhancing the Efficacy of Inclisiran for Hypercholesterolemia Using Nanocarriers: In Vitro and In Silico Approaches

Abstract

Background: A novel small interfering RNA (siRNA) therapy, inclisiran, targeting PCSK9, which regulates LDL cholesterol, is a promising treatment for hypercholesterolemia. Although it is effective, the clinical use of Inclisiran is hindered because of its low bioavailability and requirement for frequent injections. Aim: The objective of these studies is to improve the therapeutic effects of Inclisiran by creating LNP-based nanocarriers that can be targeted to the liver, resulting in increased bioavailability, stability and the ability to inject less frequently. Methods: The solvent evaporation method was used to prepare LNPs containing inclisiran which were surface modified for liver-targeted delivery. The physicochemical parameters of the LNPs were evaluated, such as their size, zeta potential, EE, and morphology. The HepG2 liver cell line was used for in vitro cytotoxicity studies, and the gene silencing efficacy was also determined by RT-qPCR to determine the silencing of PCSK9 mRNA expression. Furthermore, molecular docking was used to evaluate the binding of Inclisiran with liver-specific receptors. Key Results: The inclusion of inclisiran in LNPs had an appropriate particle size, high encapsulation efficiency, and increased stability when compared to free drug. Cytotoxicity assays demonstrated that these were very little toxic and confocal microscopy confirmed efficient cellular uptake by liver cells. A significant decrease in the level of PCSK9 expression was seen by RT-qPCR analysis which was suggestive of successful gene silencing. The in vitro results were confirmed by molecular docking which demonstrated a favorable binding of Inclisiran to liver-specific receptors. Conclusion & Clinical Significance: This work shows that Inclisiran-loaded LNPs is a viable approach to improve the drug's bioavailability and liver-targeted delivery, potentially decreasing the number of doses needed. The benefits of using nanocarriers in Inclisiran therapy for hypercholesterolemia are evident in terms of better clinical utility, patient adherence and treatment efficacy.