Glutaminyl Cyclases are responsible for the formation of pyroglutamyl residues at the N-terminus of peptides out of glutamines and glutamates. Recently N-truncated forms of pyroglutamylated Aβ-peptides came into the focus of Alzheimer’s Disease (AD) research.
The reason is their enforced amyloidogenicity, likely contributing to progression of the disorder.
Inhibitors of QC are currently in clinical investigations as new therapeutic approach for AD patients. For the drug development process the binding mode of inhibitors in the active site of the target enzyme are useful tools for understanding the necessarily molecular interactions.
Hereby we characterized the binding mode of the inhibitor PBD150, an early tool compound in the drug discovery program, by applying a combined approach of in silico analysis and site directed mutagenesis.
The co-crystallized compound reveals different poses in QC’s of several origins. These poses were characterized via a semi- empirical approach based on a linear-scaling divide-and-conquer methodology, and further analyzed using the SE-COMBINE approach.
Following this approach, site directed mutagenesis was performed to replace key amino acids of the active site. The results indicate strong evidence for different binding modes of PBD150 in the crystal structures compared with that in solution.
Mirko Buchholz, PhD
Fraunhofer Institute for Cell Therapy and Immunology, Department of Drug Design and Target Validation, Germany
Mirko Buchholz, Birgit Koch, Michael Wermann, Ulrich Heiser, Stephan Schilling and Hans-Ulrich Demuth
Glutaminyl Cyclases are responsible for the formation of pyroglutamyl residues at the N-terminus of peptides out of glutamines and glutamates. Recently N-truncated forms of pyroglutamylated Aβ-peptides came into the focus of Alzheimer’s Disease (AD) research.
The reason is their enforced amyloidogenicity, likely contributing to progression of the disorder.
Inhibitors of QC are currently in clinical investigations as new therapeutic approach for AD patients. For the drug development process the binding mode of inhibitors in the active site of the target enzyme are useful tools for understanding the necessarily molecular interactions.
Hereby we characterized the binding mode of the inhibitor PBD150, an early tool compound in the drug discovery program, by applying a combined approach of in silico analysis and site directed mutagenesis.
The co-crystallized compound reveals different poses in QC’s of several origins. These poses were characterized via a semi- empirical approach based on a linear-scaling divide-and-conquer methodology, and further analyzed using the SE-COMBINE approach.
Following this approach, site directed mutagenesis was performed to replace key amino acids of the active site. The results indicate strong evidence for different binding modes of PBD150 in the crystal structures compared with that in solution.
Mirko Buchholz, PhD is the Head of the "Drug Design and Analytical Chemistry" Unit at the Fraunhofer Institute for Cell Therapy and Immunology – Department of Drug Design and Target Validation, Halle(Saale) - Germany. He is also guest Lecturer, University of Leipzig, Leipzig - Germany.
2002 masters degree in pharmacy
2007 PhD in medicinal and computational chemistry
2007 - 2013 Probiodrug AG; postdoc, senior scientist, Head of Computational Chemistry
since 10/2013 Head of the Drug Design and Analytical Chemistry Unit, Fraunhofer IZI-MWT
(1) Schilling, S.; Zeitschel, U.; Hoffmann, T.; Heiser, U.; Francke, M.; Kehlen, A.; Holzer, M.; Hutter-Paier, B.; Prokesch, M.; Windisch, M. Glutaminyl Cyclase Inhibition Attenuates Pyroglutamate Aβ And Alzheimer's Disease–Like Pathology. Nat Med 2008, 14, 1106-1111.
(2) Nussbaum, J. M.; Schilling, S.; Cynis, H.; Silva, A.; Swanson, E.; Wangsanut, T.; Tayler, K.; Wiltgen, B.; Hatami, A.; Rönicke, R.; Reymann, K.; Hutter-Paier, B.; Alexandru, A.; Jagla, W.; Graubner, S.; Glabe, C. G.; Demuth, H.; Bloom, G. S. Prion-Like Behaviour And Tau-Dependent Cytotoxicity Of Pyroglutamylated Amyloid-Β. Nature 2012, 1-5.
(3) Buchholz, M.; Heiser, U.; Schilling, S.; Niestroj, A. J.; Zunkel, K.; Demuth, H. The First Potent Inhibitors For Human Glutaminyl Cyclase: Synthesis And Structure-Activity Relationship. J Med Chem 2006, 49, 664-677.
(4) Koch, B.; Buchholz, M.; Wermann, M.; Heiser, U.; Schilling, S.; Demuth, H. Probing Secondary Glutaminyl Cyclase (QC) Inhibitor Interactions Applying An In Silico-Modeling/Site-Directed Mutagenesis Approach: Implications For Drug Development. Chemical biology & drug design 2012, 80, 937-946.