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

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

  • Mie Kristensen, Københavns Universitet
  • ,
  • Krzysztof Kucharz, Københavns Universitet
  • ,
  • Eduardo Felipe Alves Fernandes, Københavns Universitet
  • ,
  • Kristian Strømgaard, Københavns Universitet
  • ,
  • Morten Schallburg Nielsen
  • Hans Christian Cederberg Helms, Københavns Universitet
  • ,
  • Anders Bach, Københavns Universitet
  • ,
  • Malte Ulrikkaholm Tofte-Hansen, Københavns Universitet
  • ,
  • Blanca Irene Aldana Garcia, Københavns Universitet
  • ,
  • Martin Lauritzen, Københavns Universitet
  • ,
  • Birger Brodin, Københavns Universitet

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.

OriginalsprogEngelsk
Artikelnummer661
TidsskriftPharmaceutics
Vol/bind12
Nummer7
Sider (fra-til)1-24
Antal sider24
ISSN1999-4923
DOI
StatusUdgivet - 2020

Se relationer på Aarhus Universitet Citationsformater

ID: 194076481