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Protein-Templated Hit Identification through an Ugi Four-Component Reaction** |
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| Authors: | Federica Mancini Dr M Yagiz Unver Dr Walid A M Elgaher Dr Varsha R Jumde Alaa Alhayek Dr Peer Lukat Dr Jennifer Herrmann Dr Martin D Witte Dr Matthias Köck Prof?Dr Wulf Blankenfeldt Prof?Dr Rolf Müller Prof?Dr Anna K H Hirsch |
| Institution: | 1. Department for Drug Design and Optimization, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS)–, Helmholtz Centre for Infection Research (HZI), Campus Building E8.1, 66123 Saarbrücken, Germany
Department of Pharmacy, Saarland University, Campus Building E8.1, 66123 Saarbrücken, Germany These authors contributed equally to this work.;2. Department for Drug Design and Optimization, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS)–, Helmholtz Centre for Infection Research (HZI), Campus Building E8.1, 66123 Saarbrücken, Germany Stratingh Institute for Chemistry, University of Groningen, Nijenborgh 7, 9747 AG, Groningen, The Netherlands These authors contributed equally to this work.;3. Department for Drug Design and Optimization, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS)–, Helmholtz Centre for Infection Research (HZI), Campus Building E8.1, 66123 Saarbrücken, Germany;4. Department of Structure and Function of Proteins, HZI, 38124 Braunschweig, Germany;5. Department of Microbial Natural Products, HIPS–HZI, 66123 Saarbrücken, Germany;6. Stratingh Institute for Chemistry, University of Groningen, Nijenborgh 7, 9747 AG, Groningen, The Netherlands;7. Department of Structure and Function of Proteins, HZI, 38124 Braunschweig, Germany Institute for Biochemistry, Biotechnology and Bioinformatics, Technische Universität Braunschweig, Spielmannstr. 7, 38106 Braunschweig, Germany;8. Department of Pharmacy, Saarland University, Campus Building E8.1, 66123 Saarbrücken, Germany Department of Microbial Natural Products, HIPS–HZI, 66123 Saarbrücken, Germany |
| Abstract: | Kinetic target-guided synthesis represents an efficient hit-identification strategy, in which the protein assembles its own inhibitors from a pool of complementary building blocks via an irreversible reaction. Herein, we pioneered an in situ Ugi reaction for the identification of novel inhibitors of a model enzyme and binders for an important drug target, namely, the aspartic protease endothiapepsin and the bacterial β-sliding clamp DnaN, respectively. Highly sensitive mass-spectrometry methods enabled monitoring of the protein-templated reaction of four complementary reaction partners, which occurred in a background-free manner for endothiapepsin or with a clear amplification of two binders in the presence of DnaN. The Ugi products we identified show low micromolar activity on endothiapepsin or moderate affinity for the β-sliding clamp. We succeeded in expanding the portfolio of chemical reactions and biological targets and demonstrated the efficiency and sensitivity of this approach, which can find application on any drug target. |
| Keywords: | drug discovery kinetic target-guided synthesis protein–protein interaction inhibitors protein-templated reactions Ugi reaction |