Development of Brain Targeting Peptide Based MMP-9 Inhibiting Nanoparticles for the Treatment of Brain Diseases with Elevated MMP-9 Activity

Abstract

Latent and active levels of cerebral matrix metalloproteinase 9 (MMP-9) are elevated in neurological diseases and brain injuries, contributing to neurological damage and poor clinical outcomes. This study aimed developing peptide-based nanoparticles with ability to cross the blood-brain-barrier (BBB) and inhibit MMP-9.Three amphiphilic peptides were synthesised containing brain-targeting ligands (HAIYPRH or CKAPETALC) conjugated with MMP-9 inhibiting peptide (CTTHWGFTLC) linked by glycine (spacer) at the N-terminus, and the peptide sequences were conjugated at the Nterminus to cholesterol. 19 F-NMR assay was developed to measure MMP-9 inhibition. Cell toxicity was evaluated by the LDH assay, and dialysis studies were conducted with/without fetal bovine serum. An in vitro model was employed to evaluate the ability of nanoparticles crossing the BBB.The amphiphilic peptide (Cholesterol-GGGCTTHWGFTLCHAIYPRH) formed nanoparticles (average size of 202.8 nm) with ability to cross the BBB model. MMP-9 inhibiting nanoparticles were non-toxic to cells, and reduced MMP-9 activity from kobs of 4.5×10 -6 s -1 to complete inhibition. Dialysis studies showed that nanoparticles did not disassemble by extreme dilution (40 folds), but gradually hydrolysed by serum enzymes.In conclusion, the MMP-9 inhibiting nanoparticles reduced the activity of MMP-9, with acceptable serum stability, minimal cell toxicity and ability to cross the in vitro BBB model.

Greater mechanistic understanding of the cutaneous pathogenesis of Stevens–Johnson syndrome/toxic epidermal necrolysis can shed light on novel therapeutic strategies: a comprehensive review

“…Mimics of miR-229 and miR-335 have been shown to enhance wound recovery in mouse models of diabetic healing by down-regulating MMP-9 expression [ 103 ]. Application of nanoparticles delivering MMP-9 inhibitory peptidomimetics to affected areas [ 104 ], may also offer effective potential anti-MMP-9 therapies.…”

Matrix Metalloproteinase-Responsive Drug Delivery Systems

“…The nanoparticle demonstrated satisfactory stability in serum, low toxicity toward cells, and effective penetration through an in vitro model of the BBB. 54 Therefore, MMP-9 serves as targets for drugs designed to cross the BBB.…”

“…The resulting amphiphilic peptide (Cholesterol-GGGC­TTHW­GFTL­CHAI­YPRH) self-assembled into nanoparticles that were able to traverse the BBB model (average size 202.8 nm), while inhibiting MMP-9 activity. The nanoparticle demonstrated satisfactory stability in serum, low toxicity toward cells, and effective penetration through an in vitro model of the BBB . Therefore, MMP-9 serves as targets for drugs designed to cross the BBB.…”

Transferrin-Grafted Albumin Nanoparticles for the Targeted Delivery of Apocynin and Neuroprotection in an In Vitro Model of the BBB

“…A transwell migration system was used to model the blood-brain barrier in vitro and examine the ability of nanoparticles to cross the BBB. The transwell system was prepared similarly to the previously described endothelial astrocyte coculture models [35,38,39]. Evans blue dye extravasation was performed to confirm the integrity of this in vitro model [39,40].…”