title: Discovery and structural optimization of 4-(4-(benzyloxy)phenyl)-3,4-dihydropyrimidin-2(1H)-ones as RORc inverse agonists.
abstract: OBJECTIVE
  Retinoic acid receptor-related orphan nuclear receptors (RORs) are orphan nuclear receptors that show constitutive activity in the absence of ligands. Among 3 subtypes of RORs, RORc is a promising therapeutic target for the treatment of Th17-mediated autoimmune diseases. Here, we report novel RORc inverse agonists discovered through structure-based drug design.
  
  
  METHODS
  Based on the structure of compound 8, a previously described agonist of RORa, a series of 4-(4-(benzyloxy)phenyl)-3,4-dihydropyrimidin-2(1H)-one derivatives were designed and synthesized. The interaction between the compounds and RORc was detected at molecular level using AlphaScreen assay. The compounds were further examined in 293T cells transfected with RORc and luciferase reporter gene. Thermal stability shift assay was used to evaluate the effects of the compounds on protein stability.
  
  
  RESULTS
  A total of 27 derivatives were designed and synthesized. Among them, the compound 22b was identified as the most potent RORc inverse agonist. Its IC50values were 2.39 μmol/L in AlphaScreen assay, and 0.82 μmol/L in inhibition of the cell-based luciferase reporter activity. Furthermore, the compound 22b displayed a 120-fold selectivity for RORc over other nuclear receptors. Moreover, a molecular docking study showed that the structure-activity relationship was consistent with the binding mode of compound 22b in RORc.
  
  
  CONCLUSIONS
  4-(4-(Benzyloxy)phenyl)-3,4-dihydropyrimidin-2(1H)-one derivatives are promising candidates for the treatment of Th17-mediated autoimmune diseases, such as rheumatoid arthritis, psoriasis, and multiple sclerosis.
authors.affiliation: Department of Natural Medicinal Chemistry, China Pharmaceutical University, Nanjing 211198, China.|Institute of Chemical Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
authors.forename: Xi-Shan
authors.initials: XS
authors.lastname: Wu
authors.affiliation: Institute of Chemical Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.|University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Wang
authors.affiliation: Institute of Chemical Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.|Department of Bioengineering, School of Pharmaceutical Sciences, Jilin University, Changchun 130021, China.
authors.forename: Yan-Li
authors.initials: YL
authors.lastname: Xing
authors.affiliation: Institute of Chemical Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.|University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Xiao-Qian
authors.initials: XQ
authors.lastname: Xue
authors.affiliation: Institute of Chemical Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Institute of Chemical Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
authors.forename: Yong-Zhi
authors.initials: YZ
authors.lastname: Lu
authors.affiliation: Institute of Chemical Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.|Department of Bioengineering, School of Pharmaceutical Sciences, Jilin University, Changchun 130021, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Song
authors.affiliation: Institute of Chemical Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
authors.forename: Xiao-Yu
authors.initials: XY
authors.lastname: Luo
authors.affiliation: Institute of Chemical Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
authors.forename: Chun
authors.initials: C
authors.lastname: Wu
authors.affiliation: Department of Bioengineering, School of Pharmaceutical Sciences, Jilin University, Changchun 130021, China.
authors.forename: Yu-Lai
authors.initials: YL
authors.lastname: Zhou
authors.affiliation: Department of Natural Medicinal Chemistry, China Pharmaceutical University, Nanjing 211198, China.
authors.forename: Jian-Qin
authors.initials: JQ
authors.lastname: Jiang
authors.affiliation: Institute of Chemical Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Xu
mesh_terms: D001327:Autoimmune Diseases / Q000188:drug therapy / Q000276:immunology; D001555:Benzene Derivatives / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D054314:Drug Inverse Agonism; D017930:Genes, Reporter; D057809:HEK293 Cells; D006801:Humans; D049410:Luciferases, Renilla / Q000235:genetics; D062105:Molecular Docking Simulation; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3 / Q000819:agonists* / Q000235:genetics; D011744:Pyrimidinones / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D013329:Structure-Activity Relationship; D058504:Th17 Cells / Q000276:immunology
chemical_list: D001555:Benzene Derivatives; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3; D011744:Pyrimidinones; D049410:Luciferases, Renilla