Abstract
By controlling access to the brain, the blood-brain barrier (BBB) restricts the entry of proteins and potential drugs to cerebral tissues. We demonstrate here the transcytosis ability of aprotinin and peptides derived from Kunitz domains using an in vitro model of the BBB and in situ brain perfusion. Aprotinin transcytosis across bovine brain capillary endothelial cell (BBCEC) monolayers is at least 10-fold greater than that of holo-transferrin. Sucrose permeability was unaffected by high concentrations of aprotinin, indicating that transcytosis of aprotinin was unrelated to changes in the BBCEC monolayer integrity. Alignment of the amino acid sequence of aprotinin with the Kunitz domains of human proteins allowed the identification and design of a family of peptides, named Angiopeps. These peptides, and in particular Angiopep-2, exhibit higher transcytosis capacity and parenchyma accumulation than aprotinin. Overall, these results suggest that these Kunitz-derived peptides could be advantageously used as a new brain delivery system for pharmacological agents that do not readily enter the brain.
Footnotes
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This work was supported by research funding from the National Research Council Canada Industrial Research Assistance Program to Angiochem and by a grant from National Sciences and Engineering Research Council of Canada (to R.B.).
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Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
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doi:10.1124/jpet.107.131318.
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ABBREVIATIONS: BBB, blood-brain barrier; CNS, central nervous system; LRP, low-density lipoprotein receptor-related protein; RAP, receptor-associated protein; APP, amyloid precursor protein; KPI, Kunitz protease inhibitor; BBCEC, bovine brain capillary endothelial cell; BSA, bovine serum albumin; PBS, phosphate-buffered saline; HPLC, high-performance liquid chromatography; TCA, trichloroacetic acid; LDL, low-density lipoprotein; Vd, volume of distribution.
- Received September 13, 2007.
- Accepted December 20, 2007.
- The American Society for Pharmacology and Experimental Therapeutics
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