Original Article IN SILICO ANALYSIS OF TECOVIRIMAT A REPURPOSED DRUG AGAINST THE MONKEYPOX VIRUS, ITS OFF-TARGET HUMAN PROTEINS, AND IMPACT ON HUMAN HEALTH JANVI AGGARWAL 1 , POONAM SHARMA 1* Department of Zoology, Gargi College, University of Delhi, India * Corresponding author: Poonam Sharma; * Email: poonam.sharma@gargi.du.ac.in Received: 19 Aug 2023, Revised and Accepted: 04 Oct 2023 ABSTRACT Objective: In this study, in silico analysis of human off-target proteins of tecovirimat, an investigational drug reported to stop monkey pox virus infection by binding to a protein that the virus uses to enter host cells was performed to better understand its off-target long-term and short-term effects on other important biological processes in patients. Methods: The target and off-target proteins of the drug, as well as their characteristics, protein-protein interactions, and the pathways they are involved in, were thoroughly analyzed using a number of databases, including Drug Bank, the NCBI Gene Database, BLAST, the UCSC Gene Sorter, Gene MANIA, STRING, and the Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway Database. Results: The current study showed that although the repurposing drug tecovirimat aids in the treatment of patients with monkeypox by binding to the viral p37 protein, it can also accidentally interfere with vital biological processes by interacting with off-target proteins or by indirectly interfering with the proteins that interact with these target proteins. Conclusion: The findings highlight the importance of extensively assessing and evaluating all repurposed drugs for their off-target effects before making them available to the general public. Keywords: Monkeypox, Tecovirimat, p37 protein, Rheumatoid arthritis, Alzheimer's disease © 2023 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/) DOI: https://dx.doi.org/10.22159/ijap.2023v15i6.49248. Journal homepage: https://innovareacademics.in/journals/index.php/ijap INTRODUCTION Monkeypox is caused by a DNA virus (MPV, MPXV, or hMPXV), which was found in humans in 2003. Monkeypox virus is a zoonotic virus that belongs to the orthopoxvirus family and has a genome size of 190 kb. The monkeypox virus, like other poxviruses, has an oval shape and an outer lipoprotein membrane that shields viral enzymes, DNA, and transcription factors. Monkeypox viruses rely mostly on the protein encoded in their genome that permits them to multiply in the cytoplasm of the host cells [1]. Like smallpox and rabbitpox, monkeypox is a poxvirus with a peculiar life cycle. Despite being DNA viruses, monkeypox totally bypasses the host cell nucleus and instead completes replication, transcription, translation, and virus assembly in the cytoplasm. MPXV transmission can occur through an animal bite or scratch direct contact with an infected animal's blood, biological fluids, tissue, or sores. As soon as the virus attaches to and fuses with the host cell, it uncoats and begins to produce early genes. At this stage, DNA replication initiates, followed by the transcription of early and late genes. The p37 protein aids in the formation and coating of intracellular mature virions (IMVs), which can either remain inside host cells as cell-associated virus (CEV) particles or migrate outside cells as extracellular enveloped virus (EEV) particles. p37 is a 37 kDa peripheral membrane protein expressed by the F13L gene that plays an important role in IMV particle envelopment. As a result, p37 is responsible for virulence by producing and egressing wrapped virions. The emergence of zoonotic diseases has grown increasingly widespread as a result of a variety of factors such as urbanization, deforestation, tourism, zoos, climate change, and wildlife exploitation. Drug repurposing has effectively found promising candidate drugs that potentially open up new treatment paths for treating these emerging viruses such as SARS-CoV-2, MERS-CoV, and H5N1 [2]. Repurposing existing drugs can be appealing, given the process is often safer, less expensive, and can be carried out in less time. One such repurposed medicine, tecovirimat, was recently licensed by the US Food and medicine Administration (FDA) against poxviruses and targets the critical viral p37 protein [3] (fig. 1). Fig. 1: Structure of tecovirimat drug (a) 2D structure (b) 3D structure Tecovirimat inhibits the crucial viral p37 protein and consequently prevents the formation and egress of enveloped virions, which are essential for virulence (fig. 2). As most investigational drugs have a range of mild to severe short-and long-term unfavorable side effects, this study aimed to understand tecovirimat’s off-target effects by identifying and analyzing structurally comparable off-target proteins. It also aimed to explore the possibility of analyzing tecovirimat drug behavior in silico for effective pathological control and after-effect management. International Journal of Applied Pharmaceutics ISSN- 0975-7058 Vol 15, Issue 6, 2023