[en] As a wide variety of pro-inflammatory cytokines are involved in the development of rheumatoid arthritis (RA), there is an urgent need for the discovery of novel therapeutic strategies. Among these, the inhibition of the NF-kappaB inducing kinase (NIK), a key enzyme of the NF-kappaB alternative pathway activation, represents a potential interesting approach. In fact, NIK is involved downstream of many tumor necrosis factor receptors (TNFR) like CD40, RANK or LTbetaR, implicated in the pathogenesis of RA. But, up to now, the number of reported putative NIK inhibitors is extremely limited. In this work, we report a virtual screening (VS) study combining various filters including high-throughput docking using a 3D-homology model and ranking by using different scoring functions. This work led to the identification of two molecular fragments, 4H-isoquinoline-1,3-dione (5) and 2,7-naphthydrine-1,3,6,8-tetrone (6) which inhibit NIK with an IC(50) value of 51 and 90 microM, respectively. This study opens new perspectives in the field of the NF-kappaB alternative pathway inhibition.
Piette, Jacques ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Virologie - Immunologie - Département des sciences de la vie - GIGA-Research
Frederick, Raphael
Language :
English
Title :
NF-kappaB inducing kinase (NIK) inhibitors: identification of new scaffolds using virtual screening.
F. Martinon, and J. Tschopp Trends Immunol. 26 2005 447
M. Karin, and Y. Ben-Neriah Annu. Rev. Immunol. 18 2000 621
E. Dejardin Biochem. Pharmacol. 72 2006 1161
S. Vallabhapurapu, A. Matsuzawa, W. Zhang, P.H. Tseng, J.J. Keats, H. Wang, D.A. Vignali, P.L. Bergsagel, and M. Karin Nat. Immunol. 9 2008 1364
G.X. Liao, M.Y. Zhang, E.W. Harhaj, and S.C. Sun J. Biol. Chem. 279 2004 26243
E. Claudio, K. Brown, S. Park, H. Wang, and U. Siebenlist Nat. Immunol. 3 2002 958
H.J. Coope, P.G. Atkinson, B. Huhse, M. Belich, J. Janzen, M.J. Holman, G.G. Klaus, L.H. Johnston, and S.C. Ley EMBO J. 21 2002 5375
E. Dejardin, N.M. Droin, M. Delhase, E. Haas, Y. Cao, C. Makris, Z.W. Li, M. Karin, C.F. Ware, and D.R. Green Immunity 17 2002 525
Flohr, S. N., T. US 6,841,556 B2, 2005.
J. Mortier, R. Frederick, C. Ganeff, C. Remouchamps, P. Talaga, L. Pochet, J. Wouters, J. Piette, E. Dejardin, and B. Masereel Biochem. Pharmacol. 79 2010 1462
Chen, G.; Cushing, T. D.; Fisher, B.; He, X.; Li, K.; Li, Z.; McGee, L., R. WO 2009/158011 A1, 2009.
B.K. Shoichet Nature 432 2004 862
J.J. Irwin, and B.K. Shoichet J. Chem. Inf. Comput. Sci. 45 2005 177
D.C. Rees, M. Congreve, C.W. Murray, and R. Carr Nat. Rev. Drug Disc. 3 2004 660
C. Abad-Zapatero, and J.T. Metz Drug Discovery Today 10 2005 464
The ligand efficiency (LE) is the binding free energy for a ligand divided by its molecular size. LE = -ΔG/HA ≈ -RTln(IC50)/HA ≈ -0.592ln(IC50)/HA = LEexp. LE is a useful metric for measuring the impact on activity of the addition of more molecular bulk. Molecules that achieve a given potency with fewer heavy atoms are by definition more efficient.
C.H. Reynolds, B.A. Tounge, and S.D. Bembenek J. Med. Chem. 51 2008 2432
M. Congreve, G. Chessari, D. Tisi, and A.J. Woodhead J. Med. Chem. 51 2008 3661
C.A. Lipinski, F. Lombardo, B.W. Dominy, and P.J. Feeney Adv. Drug Delivery Rev. 23 1997 3
S.J. Teague, A.M. Davis, P.D. Leeson, and T. Oprea Angew. Chem., Int. Ed. 38 1999 3743
Tripos, I. SYBYL: 1699 South Hanley Rd., St. Louis, Missouri, 63144, USA.
G. Jones, P. Willett, R.C. Glen, A.R. Leach, and R. Taylor J. Mol. Biol. 267 1997 727
The theoretical ligand efficiency (LEscore) is obtained by dividing the affinity score value by the MW of the compound (LEscore = score/MW).
A.L. Hopkins, C.R. Groom, and A. Alex Drug Discovery Today 9 2004 430
This assay was performed by ProQinase using 33PanQinase technology. Inhibitory potency of staurosporine and of the 49 molecules identified from VS was evaluated at 50 μM on human recombinant NIK. DMSO was used as co-solvent and its final concentration was 1%. NIK was expressed in Sf9 insect cells as human recombinant GST-fusion protein. The kinase was purified by affinity chromatography using GSH-agarose. The purity of the kinase was checked by SDS-PAGE/silver staining and the identity was verified by mass spectroscopy. The NIK activity was measured as the incorporation on an artificial substrate of 33P produced by hydrolysis of [γ-33P]-ATP. The substrate (RBER-CHKtide) was an artificial fusion protein expressed in Escherichia coli. It was consisting of a N-terminal GSTtag separated by a thrombin cleavage site from a fragment of the human retinoblastoma protein RB1, amino acids S773-K928 followed by 11 Arg residues (ER) and a peptide sequence KKKVRSGLYRSPSMPENLNRPR (CHKtide).
J.L. Bolton, M.A. Trush, T.M. Penning, G. Dryhurst, and T.J. Monks Chem. Res. Toxicol. 13 2000 135
H.R. Tsou, M. Otteng, T. Tran, M.B. Floyd, M. Reich, G. Birnberg, K. Kutterer, S. Ayral-Kaloustian, M. Ravi, R. Nilakantan, M. Grillo, J.P. McGinnis, and S.K. Rabindran J. Med. Chem. 51 2008 3507
H.R. Tsou, X. Liu, G. Birnberg, J. Kaplan, M. Otteng, T. Tran, K. Kutterer, Z.L. Tang, R. Suayan, A. Zask, M. Ravi, A. Bretz, M. Grillo, J.P. McGinnis, S.K. Rabindran, S. Ayral-Kaloustian, and T.S. Mansour J. Med. Chem. 52 2009 2289
J.H. Zhang, T.D. Chung, and K.R. Oldenburg J. Biomol. Screening 4 1999 67
P.W. Iversen, B.J. Eastwood, G.S. Sittampalam, and K.L. Cox J. Biomol. Screening 11 2006 247
