1-(2,6-Dibenzyloxybenzoyl)-3-(9H-fluoren-9-yl)-urea: a novel cyclophilin A allosteric activator.
Cyclophilin A (CypA) plays an important role in many physiology processes and its overexpression has been involved in many diseases including immune disease, viral infection, neuro-degenerative disease, and cancer. However, the actual role of CypA in the diseases is still far from clear, and a complete understanding of CypA is necessary in order to direct more specific and effective therapeutic strategies. Based on the screening of our in-house library through the isomer-specific proteolysis method, we find a CypA activator (1-(2,6-Dibenzyloxybenzoyl)-3-(9H-fluoren-9-yl)-urea), compound 1a, which can increase CypA's PPIase activity and give allosteric behavior. The binding affinity of compound 1a to CypA has been confirmed by Fortebio's Octet RED system and the increased phosphorylation of ERK in H446 cells is observed by treatment with both compound 1a and CsA. In order to further evaluate the binding mode between the activator and CypA, the allosteric binding site and allosteric mechanism of CypA are investigated by molecular dynamics (MD) simulations in combination with mutagenesis experiments. The results show that the allosteric binding site of CypA is 7Å away from its catalytic site and is composed of Cys52, His70, His54, Lys151, Thr152 and Lys155. Compound 1a binds to the allosteric site of CypA, stabilizing the active conformation of catalytic residues, and finally promotes the catalytic efficiency of CypA. We believe our finding of the CypA allosteric activator will be used as an effective chemical tool for further studies of CypA mechanisms in diseases.
Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China.
Maosheng
M
Lv
Ting
T
Shi
Xiaona
X
Mao
Xi
X
Li
Yingyi
Y
Chen
Jin
J
Zhu
Shuaishuai
S
Ni
Xu
X
Shen
Hualiang
H
Jiang
Jian
J
Li
Jian
J
Zhang
Jin
J
Huang
D000494:Allosteric Regulation / Q000187:drug effects; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D021984:Cyclophilin A / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D004789:Enzyme Activation / Q000187:drug effects; D005449:Fluorenes / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D009154:Mutation; D011485:Protein Binding; D011487:Protein Conformation; D014508:Urea / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology*
C577496:1-(2,6-dibenzyloxybenzoyl)-3-(9H-fluoren-9-yl)-urea; D005449:Fluorenes; D014508:Urea; D021984:Cyclophilin A