Conjugation of Therapeutic PSD-95 Inhibitors to the Cell-Penetrating Peptide Tat Affects Blood–Brain Barrier Adherence, Uptake, and Permeation

Pharmaceutics

Morten Schallburg Nielsen¹  Malte Ulrikkaholm Tofte-Hansen²  Eduardo Felipe Alves Fernandes²  Blanca Irene Aldana Garcia²  Kristian Strømgaard²  Anders Bach²  Krzysztof Kucharz³  Martin Lauritzen³  Birger Brodin⁴  HansChristian Cederberg Helms⁴  Mie Kristensen⁴ 

[1] Department of Biomedicine, Aarhus University, DK-8000 Aarhus C, Denmark;Department of Drug Design and Pharmacology, University of Copenhagen, DK-2100 Copenhagen Ø, Denmark;Department of Neuroscience and Pharmacology, University of Copenhagen, DK-2200 Copenhagen N, Denmark;Department of Pharmacy, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen Ø, Denmark;

关键词: brain peptide-drug delivery;    blood–brain barrier;    cell-penetrating peptide;    Tat;    stroke treatment;   

DOI  :  10.3390/pharmaceutics12070661

来源: DOAJ

【 摘 要 】

Novel stroke therapies are needed. Inhibition of the interaction between the postsynaptic density-95 (PSD-95)/disc large/ZO-1 (PDZ) domains of PSD-95 and the N-methyl-D-aspartate (NMDA) receptor has been suggested as a strategy for relieving neuronal damage. The peptides NR2B9c and N-dimer have been designed to hinder this interaction; they are conjugated to the cell-penetrating peptide Tat to facilitate blood–brain barrier (BBB) permeation and neuronal uptake. Tat-N-dimer exhibits 1000-fold better target affinity than Tat-NR2B9c, but the same magnitude of improvement is not observed in terms of therapeutic effect. Differences in BBB permeation by Tat-NR2B9c and Tat-N-dimer may explain this difference, but studies providing a direct comparison of Tat-NR2B9c and Tat-N-dimer are lacking. The aim of the present study was therefore to compare the BBB uptake and permeation of Tat-NR2B9c and Tat-N-dimer. The peptides were conjugated to the fluorophore TAMRA and their chemical stability assessed. Endothelial membrane association and cell uptake, and transendothelial permeation were estimated using co-cultures of primary bovine brain capillary endothelial cells and rat astrocytes. In vivo BBB permeation was demonstrated in mice using two-photon microscopy imaging. Tissue distribution was evaluated in mice demonstrating brain accumulation of TAMRA-Tat (0.4% ID/g), TAMRA-Tat-NR2B9c (0.3% ID/g), and TAMRA-Tat-N-dimer (0.25% ID/g). In conclusion, we demonstrate that attachment of NR2B9c or N-dimer to Tat affects both the chemical stability and the ability of the resulting construct to interact with and permeate the BBB.

【 授权许可】

Unknown