Virofree, an Herbal Medicine-Based Formula, Interrupts the Viral Infection of Delta and Omicron Variants of SARS-CoV-2

Ly Hien Doan; Li-Wei Chu; Zi-Yi Huang; Anh Thuc Nguyen; Chia-Yin Lee; Chien-Ling Huang; Yu-Fen Chang; Wen-Yu Hsieh; Trang Thi Huyen Nguyen; Chao-Hsiung Lin; Chun-Li Su; Tsung-Hsien Chuang; Jin-Mei Lai; Feng-Sheng Wang; Chia-Jui Yang; Hui-Kang Liu; Yueh-Hsin Ping; Chi-Ying F Huang

doi:10.3389/fphar.2022.905197

Virofree, an Herbal Medicine-Based Formula, Interrupts the Viral Infection of Delta and Omicron Variants of SARS-CoV-2

Front Pharmacol. 2022 Jul 4:13:905197. doi: 10.3389/fphar.2022.905197. eCollection 2022.

Authors

Ly Hien Doan^{1

2}, Li-Wei Chu³, Zi-Yi Huang^{4

5}, Anh Thuc Nguyen^{1

6}, Chia-Yin Lee^{1

7}, Chien-Ling Huang¹, Yu-Fen Chang⁸, Wen-Yu Hsieh⁹, Trang Thi Huyen Nguyen¹, Chao-Hsiung Lin^{1

10

11}, Chun-Li Su¹², Tsung-Hsien Chuang⁷, Jin-Mei Lai¹³, Feng-Sheng Wang¹⁴, Chia-Jui Yang¹⁵, Hui-Kang Liu^{9

16

17}, Yueh-Hsin Ping^{3

18}, Chi-Ying F Huang^{1

4

6

19

20

21}

Affiliations

¹ Institute of Biopharmaceutical Sciences, College of Pharmaceutical Sciences, National Yang Ming Chiao Tung University, Taipei, Taiwan.
² Institute of Biotechnology, Vietnam Academy of Science and Technology, Hanoi, Vietnam.
³ Department and Institute of Pharmacology, College of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan.
⁴ Program in Molecular Medicine, College of Life Sciences, National Yang Ming Chiao Tung University, Taipei, Taiwan.
⁵ ASUS Intelligent Cloud Services, Taipei, Taiwan.
⁶ Taiwan National Graduate Program in Molecular Medicine, Academia Sinica, National Yang Ming Chiao Tung University, Taipei, Taiwan.
⁷ Immunology Research Center, National Health Research Institutes, Zhunan, Taiwan.
⁸ LumiSTAR Biotechnology, Inc., Taipei, Taiwan.
⁹ Division of Basic Chinese Medicine, National Research Institute of Chinese Medicine (NRICM), Ministry of Health and Welfare, Taipei, Taiwan.
¹⁰ Department of Life Sciences and Institute of Genome Sciences, College of Life Sciences, National Yang Ming Chiao Tung University, Taipei, Taiwan.
¹¹ Aging and Health Research Center, National Yang Ming Chiao Tung University, Taipei, Taiwan.
¹² Graduate Program of Nutrition Science, School of Life Science, National Taiwan Normal University, Taipei, Taiwan.
¹³ Department of Life Science, College of Science and Engineering, Fu Jen Catholic University, New Taipei City, Taiwan.
¹⁴ Department of Chemical Engineering, National Chung Cheng University, Chiayi, Taiwan.
¹⁵ Department of Internal Medicine, Far Eastern Memorial Hospital, New Taipei City, Taiwan.
¹⁶ Ph.D. Program in the Clinical Drug Development of Herbal Medicine, Taipei Medical University, Taipei, Taiwan.
¹⁷ Traditional Herbal Medicine Research Center, Taipei Medical University Hospital, Taipei, Taiwan.
¹⁸ Institute of Biophotonics, College of Biomedical Science and Engineering, National Yang Ming Chiao Tung University, Taipei, Taiwan.
¹⁹ Institute of Clinical Medicine, College of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan.
²⁰ Department of Biotechnology and Laboratory Science in Medicine, School of Biomedical Science and Engineering, National Yang Ming Chiao Tung University, Taipei, Taiwan.
²¹ Department of Biochemistry, School of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.

Abstract

Coronavirus disease 2019 (COVID-19) remains a threat with the emergence of new variants, especially Delta and Omicron, without specific effective therapeutic drugs. The infection causes dysregulation of the immune system with a cytokine storm that eventually leads to fatal acute respiratory distress syndrome (ARDS) and further irreversible pulmonary fibrosis. Therefore, the promising way to inhibit infection is to disrupt the binding and fusion between the viral spike and the host ACE2 receptor. A transcriptome-based drug screening platform has been developed for COVID-19 to explore the possibility and potential of the long-established drugs or herbal medicines to reverse the unique genetic signature of COVID-19. In silico analysis showed that Virofree, an herbal medicine, reversed the genetic signature of COVID-19 and ARDS. Biochemical validations showed that Virofree could disrupt the binding of wild-type and Delta-variant spike proteins to ACE2 and its syncytial formation via cell-based pseudo-typed viral assays, as well as suppress binding between several variant recombinant spikes to ACE2, especially Delta and Omicron. Additionally, Virofree elevated miR-148b-5p levels, inhibited the main protease of SARS-CoV-2 (M^pro), and reduced LPS-induced TNF-α release. Virofree also prevented cellular iron accumulation leading to ferroptosis which occurs in SARS-CoV-2 patients. Furthermore, Virofree was able to reduce pulmonary fibrosis-related protein expression levels in vitro. In conclusion, Virofree was repurposed as a potential herbal medicine to combat COVID-19. This study highlights the inhibitory effect of Virofree on the entry of Delta and Omicron variants of SARS-CoV-2, which have not had any effective treatments during the emergence of the new variants spreading.

Keywords: ACE2 (angiotensin-converting enzyme 2); COVID-19; Delta variant (B.1.617.2); Omicron variant (SARS-CoV-2); SARS-CoV-2 variants; herbal medicine; miRNA.