China Animal Husbandry and Veterinary Medicine ›› 2024, Vol. 51 ›› Issue (1): 268-277.doi: 10.16431/j.cnki.1671-7236.2024.01.027

• Preventive Veterinary Medicine • Previous Articles     Next Articles

Study on the Antagonism of Reboxetine Mesylate Against Senecavirus in vitro

GONG Youquan1, ZHOU Xiaocui3, ZHENG Hui2,5, CHEN Feng4, CAO Zhenshan4, SHA Zhou2,5, ZHANG Hui2,5, CUI Jin2,5, WU Rui1,6, NI Bo2,5   

  1. 1. College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, China;
    2. China Animal Health and Epidemiology Center, Qingdao 266032, China;
    3. China Institute of Veterinary Drug Supervision, Beijing 100081, China;
    4. Shandong Animal Disease Prevention and Control Center, Jinan 250100, China;
    5. Key Laboratory of Animal Biosafety Risk Prevention and Control (South), Ministry of Agriculture and Rural Affairs, Qingdao 266032, China;
    6. Jiamusi University, Jiamusi 154000, China
  • Received:2023-06-21 Online:2024-01-05 Published:2023-12-27

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Abstract: 【Objective】 The aim of this study was to screen compounds with antagonistic Senecavirus A (SVA) activity from an FDA-approved drug library containing 127 compounds and studying their pathways of action.【Method】 An in vitro active compounds screening platform for SVA was established using luciferase recombinant Senecavirus (rSVA-NLuc) combined with Fluc high-throughput screening technology.The compounds were screened from the FDA-approved drug library for their luciferase inhibitory activity at a concentration of 10 μmol/L.The inhibitory activity was further verified by Real-time quantitative RT-PCR, and the maximum non-toxic concentration was determined by a cytotoxicity assay which was measured by the released lactate dehydrogenase (LDH) from cell.According to the four main processes of the virus infection cycle, such as adsorption, endocytosis, replication, assembly and release, different cell treatment methods and Real-time quantitative RT-PCR, indirect immunofluorescence assay(IFA), Western blotting and viral titer assay (TCID50) were used to study the antagonistic mechanism of the screeded compound.【Result】 Eight candidate anti-SVA active molecules were screened from the FDA drug library containing 127 molecules, and one safe and effective compound, reboxetine mesylate, was identified by Real-time quantitative RT-PCR and cytotoxicity assay.Within 36 hours of virus infection in cells, reboxetine mesylate significantly reduced the expression of SVA VP3 protein and the SVA titer (P<0.01).The treatment of golden hamster kidney cells (BSR-T7/5) with reboxetine mesylate could reduce the adsorption and entry of SVA.Real-time quantitative RT-PCR also showed that reboxetine mesylate could inhibit the assembly stage of SVA, but it had no effect on the replication and release stage of SVA.【Conclusion】 This experiment screened a compound with low cytotoxicity and excellent SVA antagonistic effect from the FDA-approved drug library:Riboxetine mesylate.This compound fighted SVA infection by inhibiting the adsorption, entry, and assembly stages of SVA.This study provided important references for the further development of anti-SVA drugs.

Key words: Senecavirus A (SVA); FDA-approved drug library; high-throughput screening

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