Design, stability and efficacy of a new targeting peptide for nanoparticulate drug delivery to SH-SY5Y neuroblastoma cells

Abstract

In recent years, rabies virus-derived peptide (RDP) has shown promise as a specific neural cell targeting ligand, however stability of the peptide in human serum was unknown. Herein, we report the molecular modelling and design of an optimised peptide sequence based on interactions of RDP with the 7 subunit of the nicotinic acetylcholine receptor (nAChR). The new sequence, named DAS, designed around a 5-mer sequence which demonstrated optimal nAChR binding in silico, showed greatly improved stability for up to 8 hours in human serum in comparison to RDP, which degraded within 2 hours at 37 °C. In vitro analysis using SH-SY5Y neuroblastoma cells showed that DAS-conjugated nanoparticles containing the cytotoxic drug doxorubicin (DAS-Dox-NP) displayed significantly enhanced cytotoxicity compared with untargeted doxorubicin-loaded nanoparticles (Dox-NP). DAS-Dox-NP had no significant effect on non-neural cell types, confirming its neural-specific targeting properties. This is the first time that In this manuscript, we report the design and testing of an optimised peptide ligand, conjugated to a nanoparticulate delivery vehicle and specifically targeted to neural cells., has been reported. Future impact of an innovative targeting peptide ligand combining the ability to selectively identify the target and facilitate cellular internalisation could enable the successful treatment of many neural cell disorders.

Non-Invasive, Targeted Nanoparticle-Mediated Drug Delivery across a Novel Human BBB Model

“…To test DAS’s ability to transport a large hydrophilic compound across the BBB, DAS was conjugated to Fitc-CM-dextran and tested alongside Fitc-CM-dextran on our optimal model with results shown in Figure 8 and Figure 9 . DAS is an 18 amino acid ligand, which has an affinity for alpha 7 nicotinic acetylcholine receptors (α-7 nAChR) [ 37 ] found on endothelial cells of the BBB. It allows NP conjugated to the peptide to be transported transcellularly by RMT and is, therefore, an ideal mode to transport large hydrophilic molecules (such as antibodies) into the brain [ 7 , 22 , 23 , 24 ].…”

“…We have shown this by encapsulating Fitc-dextran, a large hydrophilic molecule (70,000 Da), within a PLGA NP and conjugating DAS peptide to the surface of the NP. DAS (NH 2 -GGGGSGCLRVGGRrRrRr-COOH) is a ligand that we previously designed [ 37 ] which aids in the transportation of the loaded NP across the BBB by RMT, due to the DAS affinity for alpha 7 nicotinic acetylcholine receptors (α-7 nAChR), which are found on the membranes of the BEC of the BBB.…”

Application progress of RVG peptides to facilitate the delivery of therapeutic agents into the central nervous system

“…Receptor-mediated endocytosis is the mechanism for RVG-carrier to penetrate through the BBB modified with RVG nanocarriers and primarily bind to nAChR 26 or γ-aminobutyric acid (GABA) receptor, 16,27 which is widely located on the extracellular surface of target cells, such as microvascular endothelial cells and neurons. 28 Then transcytosis processes were initiated, and therapeutic agents were finally released in the CNS.…”

Neuroblastoma‐targeted nanoparticles and novel nanotechnology‐based treatment methods

“…These NPs must be designed by using surface glycoproteins of the rabies virus to be able to recognize and target nAChRs. These types of drug delivery systems have shown good results in clinical trials (Corringer & Changeux, 2008; Huey, Rathbone, McCarron, & Hawthorne, 2019; Rodriguez‐Nogales et al, 2019).…”