Protein-Templated Hit Identification through an Ugi Four-Component Reaction**

Federica Mancini, M. Yagiz Unver, Walid A.M. Elgaher, Varsha R. Jumde, Alaa Alhayek, Peer Lukat, Jennifer Herrmann, Martin D. Witte, Matthias Köck, Wulf Blankenfeldt, Rolf Müller, Anna K.H. Hirsch*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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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.

Original languageEnglish
Pages (from-to)14585-14593
Number of pages9
JournalChemistry - A European Journal
Issue number64
Publication statusPublished - 17-Nov-2020


  • drug discovery
  • kinetic target-guided synthesis
  • protein-templated reactions
  • protein–protein interaction inhibitors
  • Ugi reaction

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