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Shared PhD project – IPV 2015 - UPMC
Application before the 4th May 2015
Metal-tricarbonyl-centered β-turn mimics as inhibitors of the Keap1/Nrf2 PPI interaction
PhD supervisors: Hélène Bertrand, Clotilde Policar
Laboratoire des Biomolécules - LBM UMR7203 – équipe 1
1. Département de Chimie de l’ENS, 24 rue Lhomond, 75005 Paris
2. UPMC Campus Jussieu, tour 23-33 5ème étage EST, 4 place Jussieu 75005 Paris
tel: 01 44 27 63 29, 01 44 32 24 20
contact: [email protected]; [email protected]
PhD Co-supervisor (co-tutelle): Geoff Wells
School of Pharmacy – Pharmaceutical and Biological Chemistry Department
University College London (UCL – SOP)
29-39 Brunswick Square, London WC1N 1AX, UK
Context and Procedure
This PhD project has been pre-selected for funding in the framework of the doctoral program IPV 2015
(Interfaces pour le Vivant) of UPMC (Université Pierre et Marie Curie, Sorbonne Universités, Paris).
Candidates are invited to contact the PhD supervisors before the 4th May 2015. The retained candidate
will have to submit an application form by the 11th May and will have an interview at UPMC in Paris the
26th-27th May.
Project
This project aims to develop a novel approach to target protein-protein interactions (PPIs) in a biomedical
context. It will be conducted within the Laboratoire des Biomolécules (LBM, UMR7203, Paris) and the UCL
School of Pharmacy (G. Wells’s group, University College London, UK). This PhD project will allow the
candidate to develop skills and knowledge in molecular modelling, organic synthesis, solution structure
studies, and evaluation of in vitro biological activity using biophysical techniques and cell-based imaging
and assays. Mainly located in Paris, the candidate will work in the UK lab during short stays.
In this project, we propose to use a metal tricarbonyl (Re(CO)3) motif as a tool for the control and
stabilization of β-turn structures for the design of PPIs inhibitors that can be imaged in live cells. The
Re(CO)3 moieties will serve two goals: i) they will act as structural constraints to stabilize the secondary
structure of peptide mimics or will be used as the central scaffolds for the design of small molecules; ii) they
will enable the detection and imaging of the resulting inhibitors in cells using fluorescence and/or IR
imaging without any further labelling or modifications.1
We will target the regulatory interaction between the ubiquitination facilitator Keap1 and the transcription
factor Nrf2. The Keap1/Nrf2 pathway determines the ability of the eukaryotic cell to adapt and survive under
stress conditions. Disruption of the interaction between the protein Keap1 and the transcription factor Nrf2 is
a promising approach to enhance the expression of endogenous antioxidant and free radical detoxification
gene products regulated by Nrf2.2 Pharmacological inducers of the Keap1/Nrf2 pathway are intensively
studied as potential therapeutic agents in a variety of diseases (e.g. inflammatory conditions,
neurodegenerative diseases).3
1
a) S. Clède, F. Lambert, C. Sandt, Z. Gueroui, M. Refregiers, M.-A. Plamont, P. Dumas, A. Vessieres, C. Policar,
Chem. Commun., 2012, 48, 7729-7731 ; b) H. C. Bertrand, S. Clède, R. Guillot, F. Lambert, C. Policar, Inorg. Chem.
2014, 53, 6204
2
a) T. W. Kensler, N. Wakabayashi, S. Biswal, Annu Rev Pharmacol Toxicol, 2007, 47, 89-116; b) S. Magesh, Y.
Chen, L. Hu, Med. Res. Rev., 2012, 32 (4), 687-726
3
a) R. Hancock, H. C. Bertrand, T. Tsujita, S. Naz, A. El-Bakry, J. Laoruchupong, J. D. Hayes, G. Wells, Free Radic.
Biol. Med., 2012, 52, 444–451; b) D. A. East, F. Fagani, J. Crosby, H. C. Bertrand, M. Schaap, A. Fowkes, G. Wells,
M. Campanella, M., Chemistry & Biology, 2014, 21, 1585-1586
1
Shared PhD project – IPV 2015 - UPMC
Application before the 4th May 2015
Strategy
Two classes of compounds will be developed: peptides and small molecules. Targeted libraries of both
classes will be prepared. Molecular modelling will be used to design the structures and to ensure that a
proper three-dimensional organization of the functionalities
is achieved. Once the compounds have been synthesised,
structural studies will be conducted to examine their
preferred conformation in solution. The libraries will be
screened and evaluated in vitro for their affinity for the
human Keap1 protein and for their ability to inhibit the
Figure. Examples of complexes of interest
interaction between Keap1 and Nrf2 by competitive
fluorescence polarization (FP) and FRET-based assays. Potent inhibitors will be evaluated in cell-based
assays for their toxicity and their ability to induce Nrf2-dependent transcription in various relevant cells
lines. Potential synergistic effects of concomitant treatment with small molecule mimics of superoxide
dismutase (an Nrf2-dependent enzyme), that have been developed at LBM,4 will also be evaluated in an
HT29 cellular model relevant for inflammatory bowel diseases (IBD). The subcellular localization of the
active compounds will finally be determined using the constitutive Re moiety to provide further insights into
their biological mode of action and allow us to correlate cell penetration, localization and biological activity.
Expected results
With this combined imaging and biological activity study, we expect to identify potent and selective direct
modulators of the Keap1-Nrf2 interaction that will function as pharmacological tools for the growing Nrf2
research community and serve as potential lead compounds for development as therapeutic agents in a range
of disease states. This project will contribute to a better understanding of the interdependence between
biological activity and cell penetration/localization. We expect it will bring new prospects in the
development of metal-complex-based PPIs inhibitors as therapeutic agents.
4
A.-S. Bernard, C. Giroud, H. Y. V. Ching, A. Meunier, V. Ambike, C. Amatore, M. Guille-Collignon, F. Lemaître, C.
Policar, Dalton Trans., 2012, 41, 6399–6403
2