Fluorinated Ligands Approach for the Inhibition of Rotamases – F-LAIR

The introduction of fluorine into peptide substrates should enable:
-the stabilization of the transition state conformation,
-a better biological profile,
-the study by NMR of the catalytic mechanism of Pin1 and the screening of inhibitors.
The project also includes an in silico study of the interactions of fluorinated substrates with the active site of the enzyme.

The Pin1 protein was produced from E. Coli and purified in good yields. The production and purification of 15N-13C-labeled Pin1 allowed the complete characterization of this protein by NMR.
The flexibility of the Pin1 peptidyl-prolyl isomerase was explored using molecular dynamics simulations.
A preliminary docking study as well as previous results from the literature allowed us to identify the peptide and pseudopeptide units containing fluorinated structural motifs as candidate ligands. A first generation of ligands with these motifs was synthesized.
The affinity of the ligands towards the different Pin1 sites was measured by NMR.

Fluorinated Pin1 inhibitors are relevant anticancer candidates and will also allow the screening of other inhibitors by NMR.

Publications:
1.CF2H as a hydrogen bond donor group for the fine tuning of peptide bond geometry with difluoromethylated pseudoprolines. Malquin, N.; Rahgoshay, K.; Lensen, N.; Chaume, G.; Miclet, E.; Brigaud, T. Chem. Commun. 2019, 55, 12487-12490.

2. Enantiopure 5-CF3-Proline: Synthesis, Incorporation in Peptides, and Tuning of the Peptide Bond Geometry. Sanchez, C. A.; Gadais, C.; Chaume, G.; Girard, S.; Chelain, E.; Brigaud, T. Org. Lett. 2021, 23, 382-387.

Communications:
1. Design, synthesis and biological evaluation of fluorinated Pin1 ligands. Chiara Zanato, Soha Abou Ibrahim, Ludovic Carlier, Emeric Miclet, Grégory Chaume, Evelyne Chelain, Nathalie Lensen, Julien Pytkowicz, Thierry Brigaud., 21e Congrès du GFPP, Amboise (France), 12 – 16 Mai 2019

2. How to take full advantage of the CH2-TROSY experiment? R. Lenoir-Capello, L. Imbert, S. Abou Ibrahim Alami, L. Carlier, N. Birlirakis, C. Zanato, T. Brigaud, J. Pytkowicz, G. Chaume, C. Loison, J. Boisbouvier, E. Miclet. Conférence virtuelle du GERM 2021, 6-12 avr. 2021 En ligne (France).

3. Exploring the flexibility of Pin1 peptidyl-prolyl isomerase using molecular dynamics simulations, N. Mele, B. Iorga, 9èmes Journées de la Société Française de Chemoinformatique, 21-22 novembre 2019, Paris (France)

4. Design, synthesis and biological evaluation of fluorinated Pin1 ligands. Chiara Zanato, Soha Abou Ibrahim, Ludovic Carlier, Emeric Miclet, Grégory Chaume, Evelyne Chelain, Nathalie Lensen, Julien Pytkowicz, Thierry Brigaud
EUTOPIA Science FAIR 2021
eutopia-university.eu/2-the-world-of-life-chemistry-genetics-medicine/
Video: youtu.be/0CiowBTon3A

The incorporation of fluorine atoms into biomolecules has gained a considerable interest as they are known to modulate the properties of pharmaceuticals. Fluorine can often favorably and predictably modify their biophysical and chemical properties, such as lipophilicity, chemical and metabolic stability, reactivity and conformation. Fluorinated compounds can also be used as efficient probes for 19F NMR studies. In this project, we plan to take advantage of the properties of the fluorine atoms to develop fluorinated peptides as a new class of inhibitors of Pin1 which is involved in various diseases such as cancer. The introduction of fluorine into peptide substrates should enable (i) the stabilization of the transition state conformation, (ii) a better biological profile, (iii) the study by NMR of the ligand / Pin1 catalytic site interactions and the screening of inhibitors.
This project involves a multidisciplinary consortium of chemists expert in organic synthesis, in silico docking studies and biophysical methods such as NMR.