Proteins conjugated to poly(butyl cyanoacrylate) nanoparticles as potential neuroprotective agents

Abstract

Poly(butyl cyanoacrylate) (PBCA) nanoparticles (NPs) can penetrate blood-brain barrier providing the means for drug delivery to the central nervous system. Here, we study attachment of superoxide dismutase (SOD) and anti-glutamate N-methyl D-aspartate receptor 1 (NR1) antibody to PBCA NPs with the ultimate goal to design neuroprotective therapeutics for treatment of secondary spinal cord injury. Synthesis of monodispersed, ∼200 nm-diameter PBCA NPs was performed using polymerization at pH 2.0 with Dextran 70,000 as the stabilizer. Sulfo-HSAB spacers were used to covalently attach SOD and NR1 antibodies to the dextran-coated NPs. The prepared protein-NP conjugates possessed SOD activity and were capable of binding to rat cerebellar neurons. Thus, SOD and NR1 antibodies may be simultaneously attached to PBCA NPs while retaining at least a fraction of enzymatic activity and receptor-binding ability. The conjugates showed neuroprotective efficacy in vitro with rat cerebellar cell cultures challenged by superoxide.

Nanoparticles for Targeted Delivery of Antioxidant Enzymes to the Brain after Cerebral Ischemia and Reperfusion Injury

“…with a Teflon-coated stirring bar. 31 After overnight polymerization, the reaction was stopped by adding 1 mL of 0.1 mol/L NaOH, and the nanoparticle suspension was filtered twice through filter paper (VWR Grade 410 with 1 mm particle retention) in Buchner funnel via suction filtration to remove crystallized n-Butylcyanoacrylate. Excess of surfactant was removed by repeated centrifugation (4,500 r.c.f., 1.5 hours; Allegra 64 R centrifuge, Beckman Coulter, Brea, CA, USA).…”

“…To study effect of targeting, we also decorated the surface of these bulk-loaded SOD-PLGA NPs by anti-NR1 antibody using the same crosslinking chemistry as in our in vitro study. 31 The goal of this study was to determine whether enhanced delivery of targeted antioxidant NPs to the site of neuronal injury can reduce the degree of secondary damage.…”

Enabling dual cellular destinations of polymeric nanoparticles for treatment following partial injury to the central nervous system

“…Antibody attachment to NP was confirmed by incubating the NP preparations with a fluorescent secondary antibody (AF488) recognizing the anti-AQP4 antibody. 27,57 Fluorescence intensity of NP linked to anti-AQP4 antibody was substantially higher than that of control NP preparations (Figure 1c). 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 17 (mass of resveratrol/mass of dried NP).…”

Agile delivery of protein therapeutics to CNS

“…Similarly, Lin and colleagues reported that polysorbate 80-coated PBCA nanoparticles loaded with HRP or enhanced green fluorescent protein (EGFP) can deliver these proteins to the brain in a rat model of TBI [427]. Another study evaluated dextran and polysorbate 80-coated PBCA nanoparticles carrying covalently immobilized SOD1 and anti-glutamate N-methyl D-aspartate receptor 1 antibody [428]. These protein-PBCA conjugates were shown to prevent neuronal cell death mediated by superoxide radicals O 2 ·– toxicity in the rat cerebellar cells.…”