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Protein-Templated Hit Identification via an Ugi Four-Component Reaction

submitted on 30.01.2020, 09:06 and posted on 31.01.2020, 05:29 by Federica Mancini, M. Yagiz Unver, Walid Elgaher, Varsha R. Jumde, Alaa Alhayek, Peer Lukat, jennifer herrmann, Martin D. Witte, Matthias Köck, Wulf Blankenfeldt, rolf müller, Anna K.H. Hirsch
Kinetic target-guided synthesis represents an efficient hit-identification strategy, in which the protein assembles its own inhibitors from a pool of 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 ß-sliding clamp DnaN, respectively. Highly sensitive mass-spectrometry methods enabled monitoring of the protein-templated reaction of four 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 show low micromolar activity on endothiapepsin or moderate affinity for DnaN. 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.


German Federal Ministry for Education and Research (grant 16 GW0207K to A. K. H. H., R. M. and W. B.)

Helmholtz-Association’s Initiative and Networking Fund

The Netherlands Organisation for Scientific Research (NWO-CW, ECHO-STIP grant 717014004)

Dutch Ministry of Education, Culture, Science (gravitation program 024.001.035)

European Research Council (ERC starting grant 757913; all to A. K. H. H.)


Email Address of Submitting Author


Helmholtz Institute for Pharmaceutical Research Saarland - Helmholtz Centre for Infection Research



ORCID For Submitting Author


Declaration of Conflict of Interest

The authors have no conflict of interest