An In Vivo Approach to Structure Activity Relationship Analysis of Peptide Ligands
| Autor: | Robert A. Fey, Elias Lazarides, Mark A. Bernard, Henri C. van der Heyde, Ruben Venegas, Catherine M. Woods, Xiaomin Fan |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
Lipopolysaccharides
Phage display T7 phage Stereochemistry Recombinant Fusion Proteins Pharmaceutical Science Peptide Ligands Peptides Cyclic Rats Sprague-Dawley Bacteriophage Mice Structure-Activity Relationship Peptide Library In vivo Animals Technology Pharmaceutical Bacteriophages Computer Simulation Pharmacology (medical) Amino Acid Sequence Peptide library Peptide sequence Pharmacology chemistry.chemical_classification Mice Inbred BALB C biology Organic Chemistry Brain Blood Proteins biology.organism_classification Rats Cell biology Mice Inbred C57BL Microscopy Fluorescence chemistry Cerebrovascular Circulation Pia Mater Molecular Medicine Female Pharmacophore Cell Adhesion Molecules Oligopeptides Half-Life Biotechnology |
| Zdroj: | Pharmaceutical Research. 24:868-879 |
| ISSN: | 1573-904X 0724-8741 |
| DOI: | 10.1007/s11095-007-9238-z |
| Popis: | The goals in this study were several-fold. First, to optimize the in vivo phage display methodology by incorporating phage pharmacokinetic properties, to isolate peptides that target the brain microvasculature, and then to build focused libraries to obtain structure activity relationship information in vivo to identify the optimal targeting motif. The blood pharmacokinetics of filamentous and T7 phage were evaluated to choose the optimal platform. A randomized peptide library with a motif CX10C was constructed in T7 phage and used for in vivo panning. Focused peptide libraries around each structural element of the brain-specific peptide were constructed to perform kinetic structure activity relationship (kSAR) analysis in vivo. To determine potential function, sepsis was induced in mice by LPS administration and four hours later the effect of GST-peptide on adhesion of rhodamine-labelled lymphocytes or CFDA-labelled platelets to pial microvasculature was observed by intravital microscopy. The blood phamacokinetics of T7 was rapid (half-life of 12 min) which aids the clearance of non-specific phage. In vivo panning in brain enriched for isolates expressing the motif CAGALCY. Kinetic analysis of focused libraries built around each structural element of the peptide provided for rapid pharmacophore mapping. The computer modeling data suggested the peptide showed similarities to peptide mimetics of adhesion molecule ligands. GST-CAGALCY but not GST control protein was able to inhibit the rolling and adhesion of labeled platelets to inflamed pial vasculature. GST-CAGALCY had no effect on lymphocyte adhesion. Incorporating normal blood phamacokinetics of T7 phage into in vivo phage display improves the ability to recover targeting peptide motifs and allows effective lead optimization by kSAR. This approach led to the isolation of a brain-specific peptide, CAGALCY, which appears to function as an effective antagonist of platelet adhesion to activated pial microvasculature. |
| Databáze: | OpenAIRE |
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