title: A dual inhibitor against prolyl isomerase Pin1 and cyclophilin discovered by a novel real-time fluorescence detection method.
abstract: Pin1, a peptidyl prolyl cis/trans isomerase (PPIase), is a potential target molecule for cancer, infectious disease, and Alzheimer's disease. We established a high-throughput screening method for Pin1 inhibitors, which employs a real-time fluorescence detector. This screening method identified 66 compounds that inhibit Pin1 out of 9756 compounds from structurally diverse chemical libraries. Further evaluations of surface plasmon resonance methods and a cell proliferation assay were performed. We discovered a cell-active inhibitor, TME-001 (2-(3-chloro-4-fluoro-phenyl)-isothiazol-3-one). Surprisingly, kinetic analyses revealed that TME-001 is the first compound that exhibits dual inhibition of Pin1 (IC₅₀=6.1 μM) and cyclophilin, another type of PPIase, (IC₅₀=13.7 μM). This compound does not inhibit FKBP. This finding suggests the existence of similarities of structure and reaction mechanism between Pin1 and cyclophilin, and may lead to a more complete understanding of the active sites of PPIases.
authors.affiliation: Department of Molecular Cell Biology, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi 981-8555, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Mori
authors.forename: Masafumi
authors.initials: M
authors.lastname: Hidaka
authors.forename: Yi-Chin
authors.initials: YC
authors.lastname: Lin
authors.forename: Ibuki
authors.initials: I
authors.lastname: Yoshizawa
authors.forename: Takayoshi
authors.initials: T
authors.lastname: Okabe
authors.forename: Shinichiro
authors.initials: S
authors.lastname: Egashira
authors.forename: Hirotatsu
authors.initials: H
authors.lastname: Kojima
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Nagano
authors.forename: Mamoru
authors.initials: M
authors.lastname: Koketsu
authors.forename: Mari
authors.initials: M
authors.lastname: Takamiya
authors.forename: Takafumi
authors.initials: T
authors.lastname: Uchida
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D021983:Cyclophilins / Q000037:antagonists & inhibitors*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D005453:Fluorescence; D006367:HeLa Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D019696:Peptidylprolyl Isomerase / Q000037:antagonists & inhibitors*; D022021:Tacrolimus Binding Proteins / Q000037:antagonists & inhibitors; D013844:Thiazoles / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: C558129:2-(3-chloro-4-fluorophenyl)isothiazol-3-one; D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D013844:Thiazoles; D021983:Cyclophilins; D022021:Tacrolimus Binding Proteins; C000606846:PIN1 protein, human; D019696:Peptidylprolyl Isomerase