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Accueil > Equipes de recherche > Chimie Bio-inorganique des Dérivés Soufrés et Pharmacochimie > Thémes de recherche

Metalloenzymes - Pharmaco-Toxicological Implications

publié le , mis à jour le

1- Study of a Human Orphan Cytochrome P450: CYP 2U1. (J.-L. Boucher, L. Dhers, A. Hessani, D. Mansuy)

The human genome has shown the existence of 57 CYPs; 15 of them are "orphan" since almost nothing is known about their biological roles and biochemical characteristics. Among them, CYP 2U1 is predominantly expressed in the brain. Recent studies have shown greater expression of CYP 2U1 in breast and colorectal cancers than in normal tissues, and several mutations in CYP 2U1 are observed in hereditary spastic paraplegia. Thus, P450 2U1 appears as a new pharmacological target.
We obtained CYP 2U1 by over-expression in the yeast strain W(hR) and discovered the first exogenous substrates of CYP 2U1: the drug debrisoquine, and some analogues of terfenadone.
Using the X-ray structures of several P450s of family-2 as templates, a 3D homology model of human CYP 2U1 has been obtained and dynamic docking experiments of the derivatives in the active site of CYP 2U1 allowed us to interpret the regioselectivity of the hydroxylations. Expression of a N-truncated soluble form of CYP 2U1 in E. coli is under progress and will allow spectroscopic and crystallographic studies.
Collaborations :
P. Beaune, UMR-S1147 INSERM, Univ. Paris Descartes; O. Laprévote, UMR 8638 CNRS, Univ. Paris Descartes; F. André, URA 2096 CNRS, CEA Saclay; G. Stevanin, Institut du Cerveau et de la Moëlle Epinière, U-1127 INSERM, Paris.

2. Inhibition and Two-Photon Imaging of NO-Synthases by new NADPH analogues. (J.-L. Boucher, B. Ramassamy)

Recently, a NADPH analogue called nanotrigger (NT1) targeting the NADPH site of NOS reductase domain has been synthesized. Following photoactivation, NT1 was able to inject electrons to the FAD acceptor of NOS, thereby leading to light-induced NO formation (J. Am. Chem. Soc. 2007, 129, 2178-2186).
Based on molecular modeling experiments, we designed and synthesized a novel prototype of NOS inhibitor, NS1, bearing a phosphorylated adenosine moiety linked by an alkyl chain to a chromophore substituted with an acceptor and a donor group.
One- and two-photon experiments showed that NS1 was not fluorescent in aqueous solutions but became fluorescent once bound to NOS with selective enhancement for constitutive endothelial NOS (eNOS). NS1 inhibition of NO formation by neuronal NOS was competitive with NADPH and reversible.
Co-localization experiments showed that NS1 co-localized with eNOS in human umbilical endothelial cells.
Molecular modeling studies have suggested modifications of NS1 to get access to new nanotools with increased affinity and selectivity.
Collaborations :
A. Slama-Schwok (INRA, Jouy en Josas) ; E. Deprez (ENS Cachan) ; L. Baciou, Univ. Paris Sud Orsay ; O. Féron (UCL, Brussels) ; A.L. Tsai (Houston, USA) ; D. Stuehr (Cleveland, USA).
  • Richard MA, Hamels D, Pigeon P, Top S, Dansette PM, Lee HZ, Vessières A, Mansuy D, Jaouen G. (2015) ChemMedChem. 10(6): 981-90. doi: 10.1002/cmdc.201500075.

  • Ducassou L, Jonasson G, Dhers L, Pietrancosta N, Ramassamy B, Xu-Li Y, Loriot MA, Beaune P, Bertho G, Lombard M, Mansuy D, André F, Boucher JL. (2015) Biochim Biophys Acta. . 1850(7): 1426-37. doi: 10.1016/j.bbagen.2015.03.014.

  • Bluhm U, Boucher JL, Clement B, Girreser U, Heber D, Ramassamy, Wolschendorf U. (2015) Heterocycles. . 52: 24-39.

  • Rouaud F, Romero-Perez M, Wang H, Lobysheva I, Ramassamy B, Henry E, Tauc P, Giacchero D, Boucher JL, Deprez E, Rocchi S, Slama-Schwok A. (2014) Oncotarget. . 15;5(21):10650-64.

  • Li, Y. ; Wang, H. ; Tarus, B. ; Perez, M. R. ; Morellato, L. ; Henry, E. ; Berka, V. ; Tsai, A. L. ; Ramassamy, B. ; Dhimane, H. ; Dessy, C. ; Tauc, P. ; Boucher, J. L. ; Deprez, E. ; Slama-Schwok, A., (2012) Proc Natl Acad Sci U S A. 109 (31), 12526-31.