title: Discovery of Novel Inhibitors and Fluorescent Probe Targeting NAMPT.
abstract: Nicotinamide phosphoribosyltransferase (NAMPT) is a promising antitumor target. Novel NAMPT inhibitors with diverse chemotypes are highly desirable for development of antitumor agents. Using high throughput screening system targeting NAMPT on a chemical library of 30000 small-molecules, we found a non-fluorescent compound F671-0003 and a fluorescent compound M049-0244 with excellent in vitro activity (IC50: 85 nM and 170 nM respectively) and anti-proliferative activity against HepG2 cells. These two compounds significantly depleted cellular NAD levels. Exogenous NMN rescued their anti-proliferative activity against HepG2 cells. Structure-activity relationship study proposed a binding mode for NAMPT inhibitor F671-0003 and highlighted the importance of hydrogen bonding, hydrophobic and π-π interactions in inhibitor binding. Imaging study provided the evidence that fluorescent compound M049-0244 (3 μM) significantly stained living HepG2 cells. Cellular fluorescence was further verified to be NAMPT dependent by using RNA interference and NAMPT over expression transgenic mice. Our findings provide novel antitumor lead compounds and a "first-in-class" fluorescent probe for imaging NAMPT.
authors.affiliation: Department of Pharmacology, Second Military Medical University, Shanghai, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Pharmacology, Second Military Medical University, Shanghai, China.
authors.forename: Tian-Ying
authors.initials: TY
authors.lastname: Xu
authors.affiliation: Department of Pharmacology, Second Military Medical University, Shanghai, China.
authors.forename: Xin-Zhu
authors.initials: XZ
authors.lastname: Liu
authors.affiliation: Department of Pharmacology, Second Military Medical University, Shanghai, China.
authors.forename: Sai-Long
authors.initials: SL
authors.lastname: Zhang
authors.affiliation: Department of Pharmacology, Second Military Medical University, Shanghai, China.
authors.forename: Pei
authors.initials: P
authors.lastname: Wang
authors.affiliation: Department of Pharmacology, Second Military Medical University, Shanghai, China.
authors.forename: Zhi-Yong
authors.initials: ZY
authors.lastname: Li
authors.affiliation: Department of Pharmacology, Second Military Medical University, Shanghai, China.
authors.forename: Yun-Feng
authors.initials: YF
authors.lastname: Guan
authors.affiliation: Department of Pharmacology, Second Military Medical University, Shanghai, China.
authors.forename: Shu-Na
authors.initials: SN
authors.lastname: Wang
authors.affiliation: Department of Medicinal Chemistry, Second Military Medical University, Shanghai, China.
authors.forename: Guo-Qiang
authors.initials: GQ
authors.lastname: Dong
authors.affiliation: 1] Key Laboratory of Food Safety Research, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. [2] Key Laboratory of Food Safety Risk Assessment, Ministry of Health, Beijing 100021, China.
authors.forename: Shu
authors.initials: S
authors.lastname: Zhuo
authors.affiliation: 1] Key Laboratory of Food Safety Research, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. [2] Key Laboratory of Food Safety Risk Assessment, Ministry of Health, Beijing 100021, China.
authors.forename: Ying-Ying
authors.initials: YY
authors.lastname: Le
authors.affiliation: Department of Medicinal Chemistry, Second Military Medical University, Shanghai, China.
authors.forename: Chun-Quan
authors.initials: CQ
authors.lastname: Sheng
authors.affiliation: Department of Pharmacology, Second Military Medical University, Shanghai, China.
authors.forename: Chao-Yu
authors.initials: CY
authors.lastname: Miao
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001549:Benzamides / Q000737:chemistry* / Q000494:pharmacology*; D016207:Cytokines / Q000037:antagonists & inhibitors* / Q000737:chemistry; D055808:Drug Discovery; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005456:Fluorescent Dyes / Q000737:chemistry*; D056945:Hep G2 Cells; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D054409:Nicotinamide Phosphoribosyltransferase / Q000037:antagonists & inhibitors* / Q000737:chemistry; D011810:Quinoxalines / Q000737:chemistry* / Q000494:pharmacology*; D054852:Small Molecule Libraries; D013329:Structure-Activity Relationship
chemical_list: D000970:Antineoplastic Agents; D001549:Benzamides; D016207:Cytokines; D004791:Enzyme Inhibitors; C000609445:F671-0003; D005456:Fluorescent Dyes; C000609446:M049-0244; D011810:Quinoxalines; D054852:Small Molecule Libraries; D054409:Nicotinamide Phosphoribosyltransferase; C085455:nicotinamide phosphoribosyltransferase, human; C495867:nicotinamide phosphoribosyltransferase, mouse