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Identification of peptide ligands for targeting to the blood-brain barrier.

van Rooy I, Cakir-Tascioglu S, Couraud PO, Romero IA, Weksler B, Storm G, Hennink WE, Schiffelers RM, Mastrobattista E - Pharm. Res. (2010)

Bottom Line: Binding to hCMEC/D3 cells was dose dependent.When compared to lung perfusion, phage showed an 8.5-fold (GYR) and 48-fold (GLA) preference for brain over lung compared to the control.These results indicate that two new peptide ligands have been identified that may be used for specific targeting of drugs to the blood-brain barrier.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences (UIPS), Utrecht University, P.O. Box 80082, 3508 TB, Utrecht, The Netherlands. i.vanrooy@uu.nl

ABSTRACT

Purpose: Transport of drugs to the brain is limited by the blood-brain barrier. New, specific brain endothelium ligands can facilitate brain-specific delivery of drugs.

Methods: We used phage display in an in situ brain perfusion model to screen for new brain endothelium peptide ligands.

Results: Two phage clones, displaying 15 amino acid-peptides (GLA and GYR) that were selected for brain binding in the mouse model, showed significant binding to human brain endothelium (hCMEC/D3), compared to a random control phage. This binding was not seen for other human endothelial cells (HUVEC). Binding to hCMEC/D3 cells was dose dependent. When phage GLA and GYR were individually perfused through the murine brain, their ability to bind to the brain was 6-fold (GLA) and 5-fold (GYR) higher than the control phage. When compared to lung perfusion, phage showed an 8.5-fold (GYR) and 48-fold (GLA) preference for brain over lung compared to the control.

Conclusions: These results indicate that two new peptide ligands have been identified that may be used for specific targeting of drugs to the blood-brain barrier.

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Related in: MedlinePlus

Recovery of phage GLA, GYR, and control phage RVR from whole murine brains after phage perfusion and washing. Recovery is expressed as a percentage of the input. Mean±S.D., n = 3. One way ANOVA after log transformation to correct for non-Gaussian distributions with Dunnet’s correction for multiple comparisons against control. GLA and GYR vs RVR p < 0.01.
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Fig5: Recovery of phage GLA, GYR, and control phage RVR from whole murine brains after phage perfusion and washing. Recovery is expressed as a percentage of the input. Mean±S.D., n = 3. One way ANOVA after log transformation to correct for non-Gaussian distributions with Dunnet’s correction for multiple comparisons against control. GLA and GYR vs RVR p < 0.01.

Mentions: To confirm their ability to bind to brain endothelium in situ, GLA, GYR, and random control phage RVR were individually perfused through the brain again in the in situ perfusion model. After perfusion and washing, phage were recovered from the brain, and the percentages of the input were determined (Fig. 5). Phage GYR showed a 5.0 times higher affinity to the brain than random control phage RVR. Phage GLA showed a 5.9 times higher affinity. This confirms that brain-binding phage were identified. Although the recovery percentages seem low, they are comparable to in vivo phage recovery percentages found by others (39).Fig. 5


Identification of peptide ligands for targeting to the blood-brain barrier.

van Rooy I, Cakir-Tascioglu S, Couraud PO, Romero IA, Weksler B, Storm G, Hennink WE, Schiffelers RM, Mastrobattista E - Pharm. Res. (2010)

Recovery of phage GLA, GYR, and control phage RVR from whole murine brains after phage perfusion and washing. Recovery is expressed as a percentage of the input. Mean±S.D., n = 3. One way ANOVA after log transformation to correct for non-Gaussian distributions with Dunnet’s correction for multiple comparisons against control. GLA and GYR vs RVR p < 0.01.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2837178&req=5

Fig5: Recovery of phage GLA, GYR, and control phage RVR from whole murine brains after phage perfusion and washing. Recovery is expressed as a percentage of the input. Mean±S.D., n = 3. One way ANOVA after log transformation to correct for non-Gaussian distributions with Dunnet’s correction for multiple comparisons against control. GLA and GYR vs RVR p < 0.01.
Mentions: To confirm their ability to bind to brain endothelium in situ, GLA, GYR, and random control phage RVR were individually perfused through the brain again in the in situ perfusion model. After perfusion and washing, phage were recovered from the brain, and the percentages of the input were determined (Fig. 5). Phage GYR showed a 5.0 times higher affinity to the brain than random control phage RVR. Phage GLA showed a 5.9 times higher affinity. This confirms that brain-binding phage were identified. Although the recovery percentages seem low, they are comparable to in vivo phage recovery percentages found by others (39).Fig. 5

Bottom Line: Binding to hCMEC/D3 cells was dose dependent.When compared to lung perfusion, phage showed an 8.5-fold (GYR) and 48-fold (GLA) preference for brain over lung compared to the control.These results indicate that two new peptide ligands have been identified that may be used for specific targeting of drugs to the blood-brain barrier.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences (UIPS), Utrecht University, P.O. Box 80082, 3508 TB, Utrecht, The Netherlands. i.vanrooy@uu.nl

ABSTRACT

Purpose: Transport of drugs to the brain is limited by the blood-brain barrier. New, specific brain endothelium ligands can facilitate brain-specific delivery of drugs.

Methods: We used phage display in an in situ brain perfusion model to screen for new brain endothelium peptide ligands.

Results: Two phage clones, displaying 15 amino acid-peptides (GLA and GYR) that were selected for brain binding in the mouse model, showed significant binding to human brain endothelium (hCMEC/D3), compared to a random control phage. This binding was not seen for other human endothelial cells (HUVEC). Binding to hCMEC/D3 cells was dose dependent. When phage GLA and GYR were individually perfused through the murine brain, their ability to bind to the brain was 6-fold (GLA) and 5-fold (GYR) higher than the control phage. When compared to lung perfusion, phage showed an 8.5-fold (GYR) and 48-fold (GLA) preference for brain over lung compared to the control.

Conclusions: These results indicate that two new peptide ligands have been identified that may be used for specific targeting of drugs to the blood-brain barrier.

Show MeSH
Related in: MedlinePlus