Chemical Synthesis of TFF3 Reveals Novel Mechanistic Insights and a Gut-Stable Metabolite

Nayara Braga Emidio (1) , Rajeshwari Meli (2) , Hue Tran (1) , Hayeon Baik (2) , Séverine Morisset-Lopez (3) , Alysha Elliott (1) , Mark Blaskovich (1) , Sabrina Spiller (4) , Annette Beck-Sickinger (4) , Christina Schroeder (1, 5) , Markus Muttenthaler (1, 2)

1 UQ [All campuses : Brisbane, Dutton Park Gatton, Herston, St Lucia and other locations] - The University of Queensland
2 University of Vienna [Vienna]
3 CBM - Centre de biophysique moléculaire
4 Leipzig University / Universität Leipzig
5 NIH - National Institutes of Health [Bethesda, MD, USA]

TFF3 regulates essential gastro- and neuroprotective functions, but its molecular mode of action remains poorly understood. Synthetic intractability and lack of reliable bioassays and validated receptors are bottlenecks for mechanistic and structure-activity relationship studies. Here, we report the chemical synthesis of TFF3 and its homodimer via native chemical ligation followed by oxidative folding. Correct folding was confirmed by NMR and circular dichroism, and TFF3 and its homodimer were not cytotoxic or hemolytic. TFF3, its homodimer, and the trefoil domain (TFF310-50) were susceptible to gastrointestinal degradation, revealing a gut-stable metabolite (TFF37-54; t1/2 > 24 h) that retained its trefoil structure and antiapoptotic bioactivity. We tried to validate the putative TFF3 receptors CXCR4 and LINGO2, but neither TFF3 nor its homodimer displayed any activity up to 10 μM. The discovery of a gut-stable bioactive metabolite and reliable synthetic accessibility to TFF3 and its analogues are cornerstones for future molecular probe development and structure-activity relationship studies.

Domaines: Sciences du Vivant [q-bio]

Nayara Braga Emidio, Rajeshwari Meli, Hue Tran, Hayeon Baik, Séverine Morisset-Lopez, et al.. Chemical Synthesis of TFF3 Reveals Novel Mechanistic Insights and a Gut-Stable Metabolite. Journal of Medicinal Chemistry, 2021, 64 (13), pp.9484-9495. ⟨10.1021/acs.jmedchem.1c00767⟩. ⟨hal-03335490⟩