Enhancement of Drug Absorption through the Blood−Brain Barrier and Inhibition of Intercellular Tight Junction Resealing by E-Cadherin Peptides

Abstract

E-cadherin-mediated cell-cell interactions in the zonula adherens play an important role in the formation of the intercellular tight junctions found in the blood-brain barrier. However, it is also responsible for the low permeation of drugs into the brain. In this study, HAV6 peptide derived from the EC1 domain of E-cadherin was found to enhance the permeation of 14 C-mannitol and [ 3 H(G)]-daunomycin through the blood brain barrier of the in situ rat brain perfusion model. In addition, HAV6 peptide and verapamil have a synergistic effect in enhancing the BBB permeation of daunomycin. A new intercellular-junction resealing assay was also developed using Caco-2 monolayers to evaluate new peptides (BLG2, BLG3, and BLG4) derived from the bulge regions of the EC2, EC3, and EC4 domains of E-cadherin. BLG2 and BLG4 peptides but not BLG3 peptides were found to be effective in blocking the resealing of the intercellular junctions. The positive control peptides (ADT10, ADT6, and HAV10) block the resealing of the intercellular junctions in a concentration-dependent manner. All these findings suggest that E-cadherin-derived peptides can block E-cadherin-mediated cell-cell interactions. These findings demonstrate that cadherin peptides may offer a useful targeted permeation enhancement of therapeutic agents such as anticancer drugs into the brain. KeywordsE-cadherin; cell-cell adhesion; HAV peptides; ADT peptides; intercellular junctions; adherens junction; Caco-2 cell monolayers Many drug molecules have difficulty in crossing the intestinal mucosa and the blood-brain barrier (BBB).1 , 2 For proteins and peptides, their size and hydrophilicity prevent them from crossing these biological barriers. They cannot penetrate the paracellular pathway due to the presence of intercellular tight junctions.3 Intercellular tight junctions are circumferential zipper-like seals between adjacent endothelial cells of the BBB. The tight junctions have multiple functions, including maintaining cell polarity to prevent the mixing of membrane proteins between the apical and basolateral membranes4 and functioning as a gate to control the paracellular passage of ions and solutes.5 The adheren junctions consist of major transmembrane proteins called classical cadherins (i.e., E-cadherin), which generate intercellular contacts through trans-pairing between cadherins on opposing cells.6 , 7 The * To whom correspondence should be addressed Dr. Teruna J. Siahaan, Department of Pharmaceutical Chemistry, The University of Kansas, 2095 Constant Ave., Lawrence, KS 66047, Phone: 785-864-7327, Fax: 785-864-5736, siahaan@ku.edu. NIH Public Access Author ManuscriptMol Pharm. Author manuscript; available in PMC 2012 February 7. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript tight junctions of BBB begin to restrict the diffusion of molecules with molecular weight higher than 180 Da and there is a relationship between molecular size and brain absorption.5 Although small hydrophobic anticancer agents can readily partition i...

Magnetic field enhanced convective diffusion of iron oxide nanoparticles in an osmotically disrupted cell culture model of the blood–brain barrier

“…28 Several disruption methods have been developed. These include pharmacological disruption via the administration of compounds such as cadherin peptides 29 and phospholipids, 30 infusion of hyperosmotic agents such as mannitol, 31 and physical disruption via the application of techniques such as ultrasound. 32 Transient opening of the BBB with either osmotic disruption 33 or focused ultrasound 34 has been used previously to increase IONP deposition in the brain.…”

Nanobiotechnology-based delivery strategies: New frontiers in brain tumor targeted therapies

“…Synthetic peptides based on the HAV region sequence have been studied with promising results. HAV peptides showed enhanced brain delivery of 14 C-mannitol and 3 Hdaunomycin in an in situ rat brain perfusion model [158], and have been shown to increase BBB permeability for both small and large molecular weight agents without any disruption in cerebral blood flow [159].…”

Studi Interaksi antara Segmen Dimer Kitosan dengan Peptida Ac-CA-NH2 dan Ac-TP-NH2 secara Komputasi Ab-Initio

“…ADTC1 (Ac-CADTPPVC-NH2) adalah salah satu peptida dari dua kelompok besar peptida ADT-6 dan HAV-6, Gambar 4, yang dirancang berdasarkan daerah groove dan groove EC-1 [23]. Hasil penelitian menunjukan bahwa turunan peptida ADT-6 dan HAV-6, dan salah satunya adalah ADTC1 (Ac-CADTPPVC-NH2), dapat mengganggu ikatan homofilik antara E-cadherin yang mengakibatkan meningkatkan permeabilitas parasellular pada sel epitel [24]. Modulasi integritas persimpangan interselular oleh peptida cadherin tergantung pada jenis sel epitel: Madin-Darby canine kidney (MDCK) monolayer sel lebih sensitif untuk efek HAV-6 dari sel Caco-2.…”

“…Selulosa dan kitin dan kitosan mempunyai struktur rantai utama yang sama dan hanya berbeda pada gugus fungsi yang terikat pada C2 masing-masing -OH, -NH(COOH), dan -NH2. Hasil penelitian ini diharapkan juga dapat menjelaskan mekanisme pengaruh fosfat pada modulus Young tulang buatan berbasis matriks polimer kitin dan selulosa [27], dan bahkan untuk menjelaskan mekanisme peningkatan porositas BBB dengan peptida [23][24][25][26].…”