1. Blood-brain barrier and ADME properties of selected DALDA-peptide derivatives Sylvia Van Dorpe 1, Evelien Wynendaele 1, Christian Burvenich 2, Alexandre Novoa 3, Dirk Tourwe 3, Steven Ballet 3 and Bart De Spiegeleer 1,* 1 Faculty of Pharmaceutical Sciences, DruQuaR group, Ghent University, Harelbekestraat 72, 9000 Gent 2 Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke 3 Faculty of sciences, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels *Corresponding author: Tel.: +32 9 264 8100; Fax: +32 9 264 8193, E-mail: Bart.DeSpiegeleer@UGent.beBart.DeSpiegeleer@UGent.be 2011-237c

2. DruQuaR, Fac. Pharm. Sciences, Ghent University Blood-brain barrierDALDA peptide derivativesBBB transport study BBB ADME ResultsDesirability functionDesirability: ResultsConclusions

3. Dmt-D(Arg)-Phe-Lys-NH 2 Dmt-D(Arg)-Phe-Nle-NH 2 Dmt-D(Cit)-Phe-Lys-NH 2 Dmt-D(Arg)-Aba-Gly-NH 2 P41 P42 P43 SB-Aba DruQuaR, Fac. Pharm. Sciences, Ghent University Blood-brain barrierDALDA peptide derivativesBBB transport study BBB ADME ResultsDesirability functionDesirability: ResultsConclusions [M+H] + [M+2H] 2+ P42 QC: confirmed by MS 2

4. INFLUX A.Multiple time regression - IV injection - Blood & brain collection - ADME: tissue collection B.Capillary depletion - Distribution EC vs. parenchyma - Density centrifugation (Dextran ) EFFLUX Brain to blood transport - ICV injection - Brain collection - Remaining activity Fat tissues Parenchyma Capillaries Parenchyma DruQuaR, Fac. Pharm. Sciences, Ghent University Blood-brain barrierDALDA peptide derivativesBBB transport study BBB ADME ResultsDesirability functionDesirability: ResultsConclusions

5. Peptide K in (µl/(g × min)) (1) Capillary fraction (%) (2) Parenchyma fraction (%) (2) k out (min -1 ) (1) P41 0.37 ± 0.0823.12 ± 10.2676.88 ± 10.260.058 ± 0.11 P42 0.25 ± 0.2532.97 ± 9.9967.03 ± 9.990.039 ± 0.11 P43 -0.31 ± 0.4519.39 ± 15.8180.61 ± 15.810.027 ± 0.022 SB-Aba 0.45 ± 0.2030.95 ± 19.7569.05 ± 19.750.096 ± 0.046 DruQuaR, Fac. Pharm. Sciences, Ghent University (1) Regression coefficient ± SEM (2) Fraction ± standard deviation Blood-brain barrierDALDA peptide derivativesBBB transport study BBB ADME ResultsDesirability functionDesirability: ResultsConclusions

6. DruQuaR, Fac. Pharm. Sciences, Ghent University P41 P42 P43 SB-Aba Blood-brain barrierDALDA peptide derivativesBBB transport study BBB ADME ResultsDesirability functionDesirability: ResultsConclusions Liver extraction

7. DruQuaR, Fac. Pharm. Sciences, Ghent University Blood-brain barrierDALDA peptide derivativesBBB transport study BBB ADME ResultsDesirability functionDesirability: ResultsConclusions (p i = 1) Influx: internal standardisation with dermorphin

8. DruQuaR, Fac. Pharm. Sciences, Ghent University PeptideD-valueRanking P410.7651 SB Aba0.7592 P420.7233 EM-20.6264 CTOP0.6125 EM-10.6106 TAPS0.6097 CTAP0.6008 Dermorphin0.5959 DAMGO0.56910 TAPP0.40911 P430.29612 Blood-brain barrierDALDA peptide derivativesBBB transport study BBB ADME ResultsDesirability functionDesirability: ResultsConclusions

9. DruQuaR, Fac. Pharm. Sciences, Ghent University Blood-brain barrierDALDA peptide derivativesBBB transport study BBB ADME ResultsDesirability functionDesirability: ResultsConclusions - BBB influx transfer structurally related to guanidine group of D-Arg (P41, P42, SB-Aba vs. P43) - Exceptional liver uptake when replacing the ε-amino group of Lys with a methyl group (P42) - BBB drugability reflected in desirability function: P41 > SB-Aba > P42 >>> dermorphin >> P43

10. Thank you for your attention!!! DruQuaR, Fac. Pharm. Sciences, Ghent University