Sorafenib-Entrapped, Self-Assembled Pullulan–Stearic Acid Biopolymer-Derived Drug Delivery System to PLC/PRF/5 Hepatocellular Carcinoma Model

Abstract

This study aimed to design a prototypic drug delivery system (DDS) made of an amphiphilic, pullulan (Pull)-derived biodegradable polymer for targeting the asialoglycoprotein receptor (ASGPR) overexpressed in HCC. Stearic acid (SA) was conjugated to increase the hydrophobicity of pullulan (Pull-SA). Methods: Pullulan (Pull) was linked to stearic acid (SA) after functional group modifications via EDC/NHS chemistry and characterized. Sorafenib tosylate (SRFT) was entrapped in pullulan-stearic acid nanoparticles (Pull-SA-SRFT) and its particle size, zeta potential, entrapment efficiency (EE), loading capacity (LC), and release efficiency was measured. The competence of Pull-SA-SRFT over SRFT in vitro was assessed using the ASGPR over-expressing PLC/PRF/5 hepatocellular carcinoma (HCC) cell line. This was done by studying cytotoxicity by MTT assay and chromosome condensation assay, early apoptosis by annexin-Pi staining, and late apoptosis by live-dead assay. The cellular uptake study was performed by incorporating coumarin-6 (C6) fluorophore in place of SRFT in Pull-SA conjugates. A biodistribution study was conducted in Swiss-albino mice to assess the biocompatibility and targeting properties of SRFT and Pull-SA-SRFT to the liver and other organs at 1, 6, 24, and 48 h. Results:The characterization studies of the copolymer confirmed the successful conjugation of Pull-SA. The self-assembled amphiphilic nanocarrier could proficiently entrap the hydrophobic drug SRFT to obtain an entrapment efficiency of 95.6% (Pull-SA-SRFT). Characterization of the synthesized nanoparticles exhibited highly desirable nanoparticle characteristics. In vitro, apoptotic studies urged that Pull-SA-SRFT nanoparticle was delivered more efficiently to HCC than SRFT. The cellular uptake study performed, gave propitious results in 4 hrs. The biodistribution study conducted in immunocompetent mice suggested that Pull-SA-SRFT was delivered more than SRFT to the liver when compared to other organs, and that the system was biocompatible. Conclusion: Pull-SA-SRFT is a promisingly safe, biodegradable, cell-specific nanocarrier and a potential candidate to target hydrophobic drugs to HCC.

The Role of Bcl-2 and Beclin-1 Complex in “Switching” between Apoptosis and Autophagy in Human Glioma Cells upon LY294002 and Sorafenib Treatment

“…Sorafenib is an inhibitor of several kinases that are involved in cancer cell proliferation and tumor angiogenesis, including Raf. Its cytotoxic effect was also evaluated in different cell lines, such as hepatoma (PLC/PRF/5) [19], hepatocellular carcinoma (Hep G2) [20], thyroid cancer (FTC133) [21], acute myeloid leukemia (HL-60, MOLM-13, HEL, and OCI-AML2) [22], and chronic myelogenous leukemia in blast phase (K-562) [23]. Decrease in the viability of these cells was also observed in our results from MOGGCCM and T98G sorafenib-treated cells.…”

The improving strategies and applications of nanotechnology-based drugs in hepatocellular carcinoma treatment

“…Researchers have prepared Pullulan self-assembled nanoparticles that target HCC cells. The particles are coupled with stearic acid to increase the hydrophobicity of the particles, are biodegradable, and could effectively capture 95.6% sorafenib ( Chirayil and Kumar, 2022 ). From the perspective of molecular mechanism, sorafenib resistance to HCC involves many regulations such as epigenetics, regulated cell death, and TME ( Tang et al, 2020 ).…”

“…Researchers have prepared Pullulan self-assembled nanoparticles that target HCC cells. The particles are coupled with stearic acid to increase the hydrophobicity of the particles, are biodegradable, and could effectively capture 95.6% sorafenib (Chirayil and Kumar, 2022).…”