title: Synthesis and inhibitory effects of novel pyrimido-pyrrolo-quinoxalinedione analogues targeting nucleoproteins of influenza A virus H1N1.
abstract: The influenza nucleoprotein (NP) is a single-strand RNA-binding protein and the core of the influenza ribonucleoprotein (RNP) particle that serves many critical functions for influenza replication. NP has been considered as a promising anti-influenza target. A new class of anti-influenza compounds, nucleozin and analogues were reported recently in several laboratories to inhibit the synthesis of influenza macromolecules and prevent the cytoplasmic trafficking of the influenza RNP. In this study, pyrimido-pyrrolo-quinoxalinedione (PPQ) analogues as a new class of novel anti-influenza agents are reported. Compound PPQ-581 was identified as a potential anti-influenza lead with EC50 value of 1 μM for preventing virus-induced cytopathic effects. PPQ produces similar anti-influenza effects as nucleozin does in influenza-infected cells. Treatment with PPQ at the beginning of H1N1 infection inhibited viral protein synthesis, while treatment at later times blocked the RNP nuclear export and the appearance of cytoplasmic RNP aggregation. PPQ resistant H1N1 (WSN) viruses were isolated and found to have a NPS377G mutation. Recombinant WSN carrying the S377G NP is resistant to PPQ in anti-influenza and RNA polymerase assays. The WSN virus with the NPS377G mutation also is devoid of the PPQ-mediated RNP nuclear retention and cytoplasmic aggregation. The NPS377G expressing WSN virus is not resistant to the reported NP inhibitors nucleozin. Similarly, the nucleozin resistant WSN viruses are not resistant to PPQ, suggesting that PPQ targets a different site from the nucleozin-binding site. Our results also suggest that NP can be targeted through various binding sites to interrupt the crucial RNP trafficking, resulting in influenza replication inhibition.
authors: Genomics Research Center, Academia Sinica, Taipei, 115, Taiwan.
authors: Meng-I
authors: MI
authors: Lin
authors: Department of Computer Science and Information Engineering, National Taiwan University, Taipei, 106, Taiwan.
authors: Bo-Han
authors: BH
authors: Su
authors: Department of Applied Chemistry, National Chiao-Tung University, 300, Taiwan.
authors: Chia-Hsin
authors: CH
authors: Lee
authors: Department of Applied Chemistry, National Chiao-Tung University, 300, Taiwan.
authors: Suz-Ting
authors: ST
authors: Wang
authors: Department of Applied Chemistry, National Chiao-Tung University, 300, Taiwan.
authors: Wen-Chun
authors: WC
authors: Wu
authors: Department of Applied Chemistry, National Chiao-Tung University, 300, Taiwan.
authors: Prasad
authors: P
authors: Dangate
authors: Genomics Research Center, Academia Sinica, Taipei, 115, Taiwan.
authors: Shi-Yun
authors: SY
authors: Wang
authors: Genomics Research Center, Academia Sinica, Taipei, 115, Taiwan.
authors: Wen-I
authors: WI
authors: Huang
authors: Genomics Research Center, Academia Sinica, Taipei, 115, Taiwan.
authors: Ting-Jen
authors: TJ
authors: Cheng
authors: Graduate Institute of Biomedical Electronics and Bioinformatics, National Taiwan University, Taipei, 106, Taiwan.
authors: Olivia A
authors: OA
authors: Lin
authors: Genomics Research Center, Academia Sinica, Taipei, 115, Taiwan.
authors: Yih-Shyun E
authors: YS
authors: Cheng
authors: Department of Computer Science and Information Engineering, National Taiwan University, Taipei, 106, Taiwan. Electronic address: yjtseng@csie.ntu.edu.tw.
authors: Yufeng Jane
authors: YJ
authors: Tseng
authors: Department of Applied Chemistry, National Chiao-Tung University, 300, Taiwan. Electronic address: cmsun@mail.nctu.edu.tw.
authors: Chung-Ming
authors: CM
authors: Sun
mesh_terms: D000998:Antiviral Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D053118:Influenza A Virus, H1N1 Subtype / Q000187:drug effects* / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D015394:Molecular Structure; D009698:Nucleoproteins / Q000378:metabolism*; D011743:Pyrimidines / Q000737:chemistry / Q000494:pharmacology*; D011758:Pyrroles / Q000737:chemistry / Q000494:pharmacology*; D011810:Quinoxalines / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D000998:Antiviral Agents; D009698:Nucleoproteins; D011743:Pyrimidines; D011758:Pyrroles; D011810:Quinoxalines; D014764:Viral Proteins
keywords: Anti-influenza; Influenza nucleoprotein; Pyrimido-pyrrolo-quinoxalinedione (PPQ); RNA-dependent RNA polymerase