Identification of the corticotropin-releasing factor receptor 1 antagonists as inhibitors of Chikungunya virus replication using a Gaussia luciferase-expressing subgenomic replicon

  • Biochem Biophys Res Commun. 2022 Dec 31;637:181-188. doi: 10.1016/j.bbrc.2022.11.013.
Yasuo Watanabe  1 ,  Youichi Suzuki  2 ,  Akino Emi  1 ,  Takeshi Murakawa  3 ,  Takayuki Hishiki  4 ,  Fumihiro Kato  5 ,  Shoichi Sakaguchi  1 ,  Hong Wu  1 ,  Takato Yano  3 ,  Chang-Kweng Lim  6 ,  Tomohiko Takasaki  7 ,  Takashi Nakano  1
Affiliations
  • 1. Department of Microbiology and Infection Control, Faculty of Medicine, Osaka Medical and Pharmaceutical University, Takatsuki, Japan.
  • 2. Department of Microbiology and Infection Control, Faculty of Medicine, Osaka Medical and Pharmaceutical University, Takatsuki, Japan. Electronic address: [email protected].
  • 3. Department of Biochemistry, Faculty of Medicine, Osaka Medical and Pharmaceutical University, Takatsuki, Japan.
  • 4. Research Center for Drug and Vaccine Development, National Institute of Infectious Diseases, Tokyo, Japan.
  • 5. Department of Virology III, National Institute of Infectious Diseases, Tokyo, Japan.
  • 6. Department of Virology I, National Institute of Infectious Diseases, Tokyo, Japan.
  • 7. Department of Virology I, National Institute of Infectious Diseases, Tokyo, Japan; BML, Inc., Tokyo, Japan.
PMID: 36403481 DOI: 10.1016/j.bbrc.2022.11.013
Abstract

The Chikungunya virus (CHIKV), an enveloped RNA virus that has been identified in over 40 countries and is considered a growing threat to public health worldwide. However, there is no preventive vaccine or specific therapeutic drug for CHIKV Infection. To identify a new inhibitor against CHIKV Infection, this study constructed a subgenomic RNA replicon expressing the secretory Gaussia luciferase (Gluc) based on the CHIKV SL11131 strain. Transfection of in vitro-transcribed replicon RNA to BHK-21 cells revealed that Gluc activity in culture supernatants was correlated with the intracellular replication of the replicon genome. Through a chemical compound library screen using the Gluc reporter CHIKV replicon, we identified several compounds that suppressed CHIKV Infection in Vero cells. Among the hits identified, CP-154,526, a non-peptide antagonist of the corticotropin-releasing factor receptor type-1 (CRF-R1), showed the strongest anti-CHIKV activity and inhibited CHIKV Infection in Huh-7 cells. Interestingly, Other CRF-R1 antagonists, R121919 and NGD 98-2, also exhibited inhibitory effects on CHIKV Infection. Time-of-drug addition and virus entry assays indicated that CP-154,526 suppressed a post-entry step of Infection, suggesting that CRF-R1 antagonists acted on a target in the intracellular replication process of CHIKV. Therefore, the Gluc reporter replicon system established in this study would greatly facilitate the development of Antiviral drugs against CHIKV Infection.

Keywords
CP-154,526; Chikungunya virus; Corticotropin-releasing factor receptor 1 antagonists; Gaussia luciferase; Inhibitor screening; Subgenomic replicon system.
Products