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Chemical modifications of proteins

​We developed original chemical and enzymatic strategies for the site-selective modification of proteins in vitro and in vivo. By relying on these approaches, we design new chemical tools such as protein-drug conjugates and chemical probes with potential diagnostic and therapeutic applications in various pathologies including cancer, inflammation and neurodegenerative diseases.

Published on 29 April 2022

​TEAM LEADER

Laurent DEVEL     

Laurent.devel@cea.fr



Our team has a long-standing experience in the conception of chemical tools to study enzymes and proteins in systems of variable complexity, including isolated targets, culture cells media and cell lysates as well as living animals. We contributed to identify several highly selective inhibitors of Matrix MetalloProteases (MMPs) (Conception of selective inhibitorRouanet-Mehouas et al J. Med. Chem. 2017, Czarny et al., J. Med. Chem. 2013, Devel et al., J. Biol. Chem 2012, Devel et al., J. Biol. Chem 2010, Devel et al., J. Biol. Chem 2006). In preclinical models, the most selective ones notably helped deciphering the essential role of those proteases in atherosclerosis, cancer progression and during viral infection (Target validation with chemical probesElla et al., J. Thorac. Cardiovasc. Surg. 2018, Marchant et al, Nat. Med. 2014, Meides et al., Int J Cancer. 2014, Johnson et al., ArteriosclerThrombVasc. Biol. 2011). Relying on the validation of this inhibitor in living animals, our team also recently designed original imaging agents to specifically detect macrophage elastase (MMP12) in preclinical models of aneurysm and atherosclerosis (Imaging agentsGona et al., J. Med. Chem 2020, Toczek et al.; J. Med. Chem 2019, Devel et al., Molecules. 2019, Razavian et al., Sci. Report 2016, Bordenave et al, Bioconjugate 2016). With the aim to document the MMPs activation status in different biological processes, we developed several series of reactive chemical probe able to covalently modify those enzymes in complex proteomes and in vivo (Activity-based probes for functional proteomic:  Kaminska et al., Angewandte Chem. Int. Ed. 2021, Torkar et al., Bioorg. Med. Chem. Lett 2013, Torkar et al., ChemBioChem 2012, Bregant et al., J. Proteomic Res. 2009, David et al., Angewandte Chem. Int. Ed. 2007). In the same vein, we are currently developing targeted proteomic approaches that combine ligand-directed chemistry and derivatization strategies with a mass tag (Sejalon-Cipolla et al., Trends in Analytical Chemistry 2021). These approaches are also used in “target fishing” projects to determine the privileged protein targets of biologically active compounds. In parallel, we explore new approaches enabling the site-specific functionalization of proteins. In this field, our recent achievements found applications in the dual radiolabeling of antibody-drug conjugates (ADCs), whose in vivo fate of both the cytotoxic payload and the protein vector can be sensitively and accurately determined by ex vivo digital imaging. Our approach completes the very restricted set of bioanalytical methods available for the detection and quantification of ADCs in various biological matrices (Cahuzac et al., Pharmaceuticals 2020).

​A NEW GENERATION OF ACTIVITY-BASED TO STUDY MMPS IN VIVO

In vivo, MMPs are subject to several post-translational events leading to their activation. In many pathophysiological processes, the role of these active forms remains difficult to understand. In particular, MMPs are differentially activated over time and some MMPs may participate in disease progression while others have protective functions. For a given pathology and according to its grade, it is therefore necessary to identify which of the overexpressed MMPs are present under their active form. To address this issue, we recently developed a new generation of chemical probes capable of selectively labeling MMPs active forms without any external trigger, which constitutes a real breakthrough in the field of activity-based probes directed against MMPs. Indeed, our strategy will enable monitoring the MMPs activation status in many preclinical models, which was impossible to achieve with the previously reported photoactivable probes.


A NEW BIOANALYTICAL STRATEGY TO DOCUMENT THE PLASMA AND TISSUES PROFILES OF BOTH COMPONENTS OF ANTIBODY-DRUG CONJUGATES

Due to their very high therapeutic potential, antibody-drug conjugates (ADCs) represent an exponentially growing class of pharmaceuticals, with a market projected to be worth more than USD 15 Billion by 2030. During the development and optimization phases of ADCs, there is a compelling need not only to design strategies for obtaining homogenous constructs but also to access robust and quantitative data regarding their pharmacokinetics and biodistribution, an essential aspect to objectivate their efficiency and eventual toxicity in preclinical models. In collaboration with G. Pieters and D. Audisio (Institut Joliot/DMTS/SCBM), we developed an original bioanalytical strategy based on site-specific dual radiolabeling of ADCs and ex vivo digital imaging, which enables monitoring simultaneously the drug (labelled with 3H) and its protein vector (labelled with 14C) during their circulation and distribution phases. This strategy, readily expandable to a wide range of protein-drug conjugates, could become unavoidable to compare the in vivo performances of conjugates varying either in their vectors, linkers or drugs, thus favoring the identification of conjugates with the highest potential for future clinical evaluations.