Viral Replication Process Research Articles

Discovery of New 1,2,4-Triazole/1,3,4-Oxadiazole-Decorated Quinolinones as Agrochemical Alternatives for Controlling Viral Infection by Inhibiting the Viral Replication and Self-Assembly Process.

Tobacco mosaic virus (TMV), a representative plant virus, is widely known and causes severe crop losses worldwide. In order to ensure the demand for crop and food security, the exploration of novel antiviral agents with outstanding activity and unique mechanisms of action is necessary. Herein, 40 new azole-quinolinone molecules were elaborately designed and systematically evaluated for their anti-TMV activity. Notably, compound A21 had significant therapeutic activity against TMV (EC50 value = 200 μg/mL), which was superior to commercial ningnanmycin (280 μg/mL). Studies on the anti-TMV mechanism showed that compound A21 could suppress the expression level of important TMV genes and affect the assembly of TMV viral particles by disrupting the self-assembly process of TMV coat protein (TMV-CP). In-depth antiviral behaviors were verified by molecular docking, fluorescence titration analysis, and TMV assembly assays, suggesting that compound A21 strongly interacted with TMV coat protein through various interactions. Overall, this promising work discloses a new paradigm for the exploitation of 2-quinolinone-based virucidal agents for hindering plant viral infection through triggering versatile antiviral behavior.

AMFR-mediated Flavivirus NS2A ubiquitination subverts ER-phagy to augment viral pathogenicity.

Flaviviruses strategically utilize the endoplasmic reticulum (ER) in their replication cycles. However, the role of ER autophagy (ER-phagy) in viral replication process remains poorly understood. Here, we reveal that prolonged Zika virus (ZIKV) infection results from the degradation of ER-phagy receptor FAM134B, facilitated by viral NS2A protein. Mechanistically, ER-localized NS2A undergoes K48-linked polyubiquitination at lysine (K) 56 by E3 ligase AMFR. Ubiquitinated NS2A binds to FAM134B and AMFR orchestrates the degradation of NS2A-FAM134B complexes. AMFR-catalyzed NS2A ubiquitination not only targets FAM134B degradation but also hinders the FAM134B-AMFR axis. Notably, a recombinant ZIKV mutant (ZIKV-NS2AK56R), lacking ubiquitination and ER-phagy inhibition, exhibits attenuation in ZIKV-induced microcephalic phenotypes in human brain organoids and replicates less efficiently, resulting in weakened pathogenesis in mouse models. In this work, our mechanistic insights propose that flaviviruses manipulate ER-phagy to modulate ER turnover, driving viral infection. Furthermore, AMFR-mediated flavivirus NS2A ubiquitination emerges as a potential determinant of viral pathogenecity.

Phyllanthus-derived Naturally-occurring Products: An Overview of their Effects against Viruses in Cell Models

Background: Infectious diseases are considered a global public health problem, with viruses being the predominant infectious agents afflicting the human population. The most used control alternatives are the search and development of vaccines and drugs. Nevertheless, their efficacy has limitations related to the immune response stimulation, resistance mechanisms, costeffort ratio, development, and production. An alternative to these drawbacks is the search for compounds isolated from plants with antiviral and/or virucidal properties. The genus Phyllanthus is a plant group producing compounds that gathers an antiviral and virucidal spectrum on different biological models. However, there is no complete review of their properties against viruses in cell models. Objective: To compile and analyse the more relevant information on the antiviral and virucidal activity in cell models, phytochemical composition, and generalities of the genus Phyllanthus. Method: The information was assembled from a general search for articles in various databanks, and the information was organized, tabulated, and discussed. Results: The taxonomic classification of the genus Phyllanthus showed discrepancies between different authors and publications. The antiviral and virucidal effects of Phyllanthus naturallyoccurring compounds on cell models showed a broad spectrum and a high chemical diversity mainly related to phenols and polyphenols. Conclusion: Antiviral and virucidal properties of Phyllanthus-derived compounds showed promising results as controlling agents against viral infections in different cell models, particularly in the viral replication and translation processes. Further studies are required to elucidate the specific mechanisms involved in these natural alternatives to expand their efficient and effective applications.

Electron Tomography as a Tool to Study SARS-CoV-2 Morphology.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a novel betacoronavirus, is the causative agent of COVID-19, which has caused economic and social disruption worldwide. To date, many drugs and vaccines have been developed for the treatment and prevention of COVID-19 and have effectively controlled the global epidemic of SARS-CoV-2. However, SARS-CoV-2 is highly mutable, leading to the emergence of new variants that may counteract current therapeutic measures. Electron microscopy (EM) is a valuable technique for obtaining ultrastructural information about the intracellular process of virus replication. In particular, EM allows us to visualize the morphological and subcellular changes during virion formation, which would provide a promising avenue for the development of antiviral agents effective against new SARS-CoV-2 variants. In this review, we present our recent findings using transmission electron microscopy (TEM) combined with electron tomography (ET) to reveal the morphologically distinct types of SARS-CoV-2 particles, demonstrating that TEM and ET are valuable tools for visually understanding the maturation status of SARS-CoV-2 in infected cells. This review also discusses the application of EM analysis to the evaluation of genetically engineered RNA viruses.

Elucidation of the Pharmacological Development and Action Mechanism of Remdesivir and Paxlovid in Combatting COVID-19

COVID-19, the disease identified in 2019 as coronavirus disease, stems from an infection by severe acute respiratory syndrome coronavirus 2, or SARS-CoV-2, which possesses a single-stranded RNA genome. Since the onset of the pandemic attributed to COVID-19, a variety of vaccines and antiviral medications have received approval from international regulatory bodies. Two notable antiviral agents, remdesivir and paxlovid, serve as treatments for viral infections. Initially developed to combat Ebola, remdesivir has also demonstrated efficacy against a broad spectrum of coronaviruses (CoV). It is a nucleoside analogue and functions by being incorporated into the RNA chain of the virus and thus induce the early termination of RNA synthesis. Paxlovid comprises two pharmacological agents, nirmatrelvir and ritonavir. The mechanism of nirmatrelvir involves the specific targeting and inhibition of the viral protease’s active site, thereby impeding the viral replication process. By inhibiting the cytochrome CYP3A4 enzyme which metabolites nirmatrelvir, ritonavir acts as a pharmacokinetic enhancer in this combination. The initial section of this review provides a comprehensive analysis of SARS-CoV-2, detailing the mechanisms of its pathogenesis and the subsequent immune responses elicited by the body. Then this article generalizes the history of development, the structure, and the mode of action of remdesivir and paxlovid.

Structural and free energy landscape analysis for the discovery of antiviral compounds targeting the cap-binding domain of influenza polymerase PB2

Influenza poses a significant threat to global health, with the ability to cause severe epidemics and pandemics. The polymerase basic protein 2 (PB2) of the influenza virus plays a crucial role in the viral replication process, making the CAP-binding domain of PB2 an attractive target for antiviral drug development. This study aimed to identify and evaluate potential inhibitors of the influenza polymerase PB2 CAP-binding domain using computational drug discovery methods. We employed a comprehensive computational approach involving virtual screening, molecular docking, and 500 ns molecular dynamics (MD) simulations. Compounds were selected from the Diverse lib database and assessed for their binding affinity and stability in interaction with the PB2 CAP-binding domain. The study utilized the generalized amber force field (GAFF) for MD simulations to further evaluate the dynamic behaviour and stability of the interactions. Among the screened compounds, compounds 1, 3, and 4 showed promising binding affinities. Compound 4 demonstrated the highest binding stability and the most favourable free energy profile, indicating strong and consistent interaction with the target domain. Compound 3 displayed moderate stability with dynamic conformational changes, while Compound 1 maintained robust interactions throughout the simulations. Comparative analyses of these compounds against a control compound highlighted their potential efficacy. Compound 4 emerged as the most promising inhibitor, with substantial stability and strong binding affinity to the PB2 CAP-binding domain. These findings suggest that compound 4, along with compounds 1 and 3, holds the potential for further development into effective antiviral agents against influenza. Future studies should focus on experimental validation of these compounds and exploration of resistance mechanisms to enhance their therapeutic utility.

Patient visit behaviour shapes the virus infection dynamics in hosts

Chronic HBV infection is a dynamic process of rapid viral replication and clearance, with large amounts of virus being produced and lost in the body daily. The patient's visit behaviour is closely related to viral load in the body, while the data-driven mathematical model still lacking. Therefore, we first include 28,912 patients to investigate the relationships between the visit behaviour and the HBV-DNA virus load. Then, we propose an HBV virus infection dynamics model that considers the patient's visit behaviour. The system is chaotic once the behaviour of patients during visits depends on the virus load. Our results may help us understand the process of viral transmission, and increasing the visit probability to the clinic can lead to stabilising HBV-DNA viral loads and further help healthcare professionals develop targeted clinical intervention strategies.

Proteomic analysis of global protein acetylation in response to WSSV infection in a crustacean Cherax quadricarinatus

Protein acetylation, a crucial post-translational modification, plays a significant role in antiviral immunity of host and viral replication processes after infection. Our previous research showed that an acetyltransferase KAT2A significantly enhanced white spot syndrome virus (WSSV) replication in hematopoietic tissue (Hpt) cells from Cherax quadricarinatus, suggesting that alterations in intracellular acetylation states affect viral infection. Nevertheless, the mechanisms by which WSSV invasion regulates intracellular protein acetylation remain unexplored. To explore the connection between protein acetylation and WSSV infection, differentially acetylated proteins (DAcPs) were identified in Hpt cells during WSSV infection by proteomic analysis. The results revealed that 25 DAcPs were identified with primarily involving in transporter activity and proton ATPase activity complex during the early stage of WSSV infection, i.e., the intracellular transport of the virions. Furthermore, that 36 DAcPs associated with chromatin assembly and acetyltransferase complex were identified in the replication stage of WSSV infection, i.e., the process of extensive viral replication. Additionally, acetylation at the K27 and K36 sites of histone H3 was strongly activated during WSSV replication; similarly, the inhibition of histone acetylation by acetyltransferase inhibiter C646 significantly suppressed WSSV replication, implying that WSSV replication requires high levels of histone acetylation, while the acetylation at the K27 and K36 sites of histone H3 probably serves as critical viral regulatory targets. This study deepens the understanding of WSSV-host interaction, and lays the groundwork for future research into the relationship between protein acetylation and viral infection in crustaceans.

Analyzing Paxlovid: Examining Its Properties, Characteristics, and Analytical and Bio‐Analytical Methods–A Comprehensive Review

ABSTRACTPaxlovid, a novel antiviral medication developed by Pfizer, has attracted considerable attention as a coronavirus disease 2019 therapeutic since it may be effective in treating the severe acute respiratory syndrome coronavirus type 2‐caused disease. Paxlovid is known as a combination therapy since it includes two agents, Nirmatrelvir and Ritonavir, which both are involved in the viral replication process inhibition. A further discussion of the molecular structure, pharmacokinetics, and pharmacodynamics of both components will help comprehend Paxlovid's mechanism of impact and effectiveness. High‐performance and spectroscopic methods, including high‐performance liquid chromatography (HPLC), infrared, and ultraviolet‐visible spectroscopy have been among the methods employed for compound identification, quantification, and structural elucidation. Bioanalytical methods have also made a significant contribution to the evaluation of Paxlovid's pharmacokinetic parameters, bioavailability, and metabolism in biological matrices. LC‐tandem mass spectrometry (LC‐MS/MS) is widely employed for the quantification of Paxlovid and its metabolites in plasma, serum, and other biological samples. This study brings forth an overview of the characteristics and properties of Paxlovid along with analytical and bioanalytical methods employed for its assessment and quantification.

Asparagine Availability Is a Critical Limiting Factor for Infectious Spleen and Kidney Necrosis Virus Replication.

Infectious spleen and kidney necrosis virus (ISKNV) has brought huge economic loss to the aquaculture industry. Through interfering with the viral replication and proliferation process that depends on host cells, its pathogenicity can be effectively reduced. In this study, we investigated the role of asparagine metabolites in ISKNV proliferation. The results showed that ISKNV infection up-regulated the expression of some key enzymes of the asparagine metabolic pathway in Chinese perch brain (CPB) cells. These key enzymes, including glutamic oxaloacetic transaminase 1/2 (GOT1/2) and malate dehydrogenase1/2 (MDH1/2) associated with the malate-aspartate shuttle (MAS) pathway and asparagine synthetase (ASNS) involved in the asparagine biosynthesis pathway, were up-regulated during ISKNV replication and release stages. In addition, results showed that the production of ISKNV was significantly reduced by inhibiting the MAS pathway or reducing the expression of ASNS by 1.3-fold and 0.6-fold, respectively, indicating that asparagine was a critical limiting metabolite for ISKNV protein synthesis. Furthermore, when asparagine was added to the medium without glutamine, ISKNV copy number was restored to 92% of that in the complete medium, indicating that ISKNV could be fully rescued from the absence of glutamine by supplementing asparagine. The above results indicated that asparagine was a critical factor in limiting the effective replication of ISKNV, which provided a new idea for the treatment of aquatic viral diseases.

African swine fever virus RNA polymerase subunits C315R and H359L inhibition host translation by activating the PKR-eIF2a pathway and suppression inflammatory responses.

ASFV C315R is homologous to the transcription factor TFIIB of large unclassified DNA viruses, and H359L is identical to the subunit 3 (RPB3) of eukaryotic RNA polymerase II. The C315R and H359L may play an important role in ASFV replication and transcription. Here, we evaluated the biological function of the C315R and H359L genes during virus replication in vitro and during infection in pigs. Results showed that C315R and H359L are highly conserved among ASFV genotype II strains; quantitative PCR (qPCR) and western blotting analyses revealed that C315R and H359L are early transcribed genes prior to viral DNA replication, but their protein expression is delayed. The immunofluorescence and western blotting analysis revealed that both proteins localized in the cell cytoplasm and nucleus at 24 h post infection, however, pH359L was mainly detected in the cell cytoplasm. Furthermore, overexpression of pH359L in MA104 cells significantly increased viral titer, RNA transcription levels, and viral protein expression levels, while overexpression of pC315R slightly enhanced ASFV replication. In contrast, siRNA targeting ASFV-H359L or C315R reduced replication efficiency in porcine macrophage culture compared to the parent ASFV-CN/SC/2019, demonstrating that C315R and H359L genes are necessary for ASFV replication. Finally, the functional role of C315R or H359L on PKR and eIF2α phosphorylation status and SG formation, as well as cytokine production were evaluated. These studies demonstrated that C315R and H359L are involved in virus replication processes in swine and play important roles in ASFV replication.

Independent repeated mutations within the alphaviruses Ross River virus and Barmah Forest virus indicates convergent evolution and past positive selection in ancestral populations despite ongoing purifying selection.

Ross River virus (RRV) and Barmah Forest virus (BFV) are arthritogenic arthropod-borne viruses (arboviruses) that exhibit generalist host associations and share distributions in Australia and Papua New Guinea (PNG). Using stochastic mapping and discrete-trait phylogenetic analyses, we profiled the independent evolution of RRV and BFV signature mutations. Analysis of 186 RRV and 88 BFV genomes demonstrated their viral evolution trajectories have involved repeated selection of mutations, particularly in the nonstructural protein 1 (nsP1) and envelope 3 (E3) genes suggesting convergent evolution. Convergent mutations in the nsP1 genes of RRV (residues 248 and 441) and BFV (residues 297 and 447) may be involved with catalytic enzyme mechanisms and host membrane interactions during viral RNA replication and capping. Convergent E3 mutations (RRV site 59 and BFV site 57) may be associated with enzymatic furin activity and cleavage of E3 from protein precursors assisting viral maturation and infectivity. Given their requirement to replicate in disparate insect and vertebrate hosts, convergent evolution in RRV and BFV may represent a dynamic link between their requirement to selectively 'fine-tune' intracellular host interactions and viral replicative enzymatic processes. Despite evidence of evolutionary convergence, selection pressure analyses did not reveal any RRV or BFV amino acid sites under strong positive selection and only weak positive selection for nonstructural protein sites. These findings may indicate that their alphavirus ancestors were subject to positive selection events which predisposed ongoing pervasive convergent evolution, and this largely supports continued purifying selection in RRV and BFV populations during their replication in mosquito and vertebrate hosts.

TDFFM: Transformer and Deep Forest Fusion Model for Predicting Coronavirus 3C-Like Protease Cleavage Sites.

COVID-19, caused by the highly contagious SARS-CoV-2 virus, is distinguished by its positive-sense, single-stranded RNA genome. A thorough understanding of SARS-CoV-2 pathogenesis is crucial for halting its proliferation. Notably, the 3C-like protease of the coronavirus (denoted as 3CLpro) is instrumental in the viral replication process. Precise delineation of 3CLpro cleavage sites is imperative for elucidating the transmission dynamics of SARS-CoV-2. While machine learning tools have been deployed to identify potential 3CLpro cleavage sites, these existing methods often fall short in terms of accuracy. To improve the performances of these predictions, we propose a novel analytical framework, the Transformer and Deep Forest Fusion Model (TDFFM). Within TDFFM, we utilize the AAindex and the BLOSUM62 matrix to encode protein sequences. These encoded features are subsequently input into two distinct components: a Deep Forest, which is an effective decision tree ensemble methodology, and a Transformer equipped with a Multi-Level Attention Model (TMLAM). The integration of the attention mechanism allows our model to more accurately identify positive samples, thus enhancing the overall predictive performance. Evaluation on a test set demonstrates that our TDFFM achieves an accuracy of 0.955, an AUC of 0.980, and an F1-score of 0.367, substantiating the model's superior prediction capabilities.

Hepatitis B Viral Protein HBx: Roles in Viral Replication and Hepatocarcinogenesis.

Hepatitis B virus (HBV) infection remains a major public health concern worldwide, with approximately 296 million individuals chronically infected. The HBV-encoded X protein (HBx) is a regulatory protein of 17 kDa, reportedly responsible for a broad range of functions, including viral replication and oncogenic processes. In this review, we summarize the state of knowledge on the mechanisms underlying HBx functions in viral replication, the antiviral effect of therapeutics directed against HBx, and the role of HBx in liver cancer development (including a hypothetical model of hepatocarcinogenesis). We conclude by highlighting major unanswered questions in the field and the implications of their answers.

Etravirine Prevents West Nile Virus and Chikungunya Virus Infection Both In Vitro and In Vivo by Inhibiting Viral Replication.

Diseases transmitted by arthropod-borne viruses such as West Nile virus (WNV) and chikungunya virus (CHIKV) pose threat to global public health. Unfortunately, to date, there is no available approved drug for severe symptoms caused by both viruses. It has been reported that reverse transcriptase inhibitors can effectively inhibit RNA polymerase activity of RNA viruses. We screened the anti-WNV activity of the FDA-approved reverse transcriptase inhibitor library and found that 4 out of 27 compounds showed significant antiviral activity. Among the candidates, etravirine markedly inhibited WNV infection in both Huh 7 and SH-SY5Y cells. Further assays revealed that etravirine inhibited the infection of multiple arboviruses, including yellow fever virus (YFV), tick-borne encephalitis virus (TBEV), and CHIKV. A deeper study at the phase of action showed that the drug works primarily during the viral replication process. This was supported by the strong interaction potential between etravirine and the RNA-dependent RNA polymerase (RdRp) of WNV and alphaviruses, as evaluated using molecular docking. In vivo, etravirine significantly rescued mice from WNV infection-induced weight loss, severe neurological symptoms, and death, as well as reduced the viral load and inflammatory cytokines in target tissues. Etravirine showed antiviral effects in both arthrophlogosis and lethal mouse models of CHIKV infection. This study revealed that etravirine is an effective anti-WNV and CHIKV arbovirus agent both in vitro and in vivo due to the inhibition of viral replication, providing promising candidates for clinical application.

A INTERAÇÃO ENTRE COVID-19 E O SISTEMA IMUNOLÓGICO: MECANISMOS E IMPLICAÇÕES CLÍNICAS

The infection by SARS-CoV-2, the virus responsible for COVID-19, triggers a series of immune responses in the host organism. These responses, while essential for pathogen elimination, can also cause significant damage to host tissues. This work details the mechanism of SARS-CoV-2 infection, from its structure to the processes of viral entry and replication, as well as the mechanisms related to infection and activation of the immune system, which can culminate in a cytokine storm. Understanding these mechanisms is crucial for developing effective therapeutic strategies, vaccines, and alternative therapies that were urgently released during the pandemic. Continuous research, especially related to the immune response, is necessary to enhance knowledge and preparedness for future pandemics.

<span>Molecular mechanism of Ensitrelvir and its similarity inhibiting SARS-CoV-2 main protease by molecular dynamics simulation</span>

The unprecedented challenge posed by the COVID-19 pandemic, driven by SARS-CoV-2, has emerged as a global threat. In response, a limited array of therapeutics has been approved for the prevention and treatment of SARS-CoV-2 infection. The main protease (Mpro) of SARS-CoV-2 has been a significant target for drug development efforts because of its crucial role in the viral replication process. This study is to investigate the efficacy of Ensitrelvir and its derivatives in inhibiting the mechanism of the Mpro target of SAR-CoV-2. Docking simulation and molecular dynamic simulation (SMD) techniques were employed for this purpose. The results indicate that the CID 166498740 derivative obtained affinity energy -9.3 kcal/mol and rupture force (Fmax) 638.3 ± 79.3 (pN), which proved that the CID 166498740 derivative strongly interacted with the Mpro target, emphasizing non-binding interactions as more crucial than hydrogen bonding in stabilizing the receptor-ligand conformation.

Genome-wide identification of ATG genes and their expression profiles under biotic and abiotic stresses in Fenneropenaeus chinensis

BackgroundAutophagy is a conserved catabolic process in eukaryotes that contributes to cell survival in response to multiple stresses and is important for organism fitness. Extensive research has shown that autophagy plays a pivotal role in both viral infection and replication processes. Despite the increasing research dedicated to autophagy, investigations into shrimp autophagy are relatively scarce.ResultsBased on three different methods, a total of 20 members of the ATGs were identified from F. chinensis, all of which contained an autophagy domain. These genes were divided into 18 subfamilies based on their different C-terminal domains, and were found to be located on 16 chromosomes. Quantitative real-time PCR (qRT-PCR) results showed that ATG genes were extensively distributed in all the tested tissues, with the highest expression levels were detected in muscle and eyestalk. To clarify the comprehensive roles of ATG genes upon biotic and abiotic stresses, we examined their expression patterns. The expression levels of multiple ATGs showed an initial increase followed by a decrease, with the highest expression levels observed at 6 h and/or 24 h after WSSV injection. The expression levels of three genes (ATG1, ATG3, and ATG4B) gradually increased until 60 h after injection. Under low-salt conditions, 12 ATG genes were significantly induced, and their transcription abundance peaked at 96 h after treatment.ConclusionsThese results suggested that ATG genes may have significant roles in responding to various environmental stressors. Overall, this study provides a thorough characterization and expression analysis of ATG genes in F. chinensis, laying a strong foundation for further functional studies and promising potential in innate immunity.

The Identification and Function of Linc01615 on Influenza Virus Infection and Antiviral Response.

Influenza virus infection poses a great threat to human health globally each year. Non-coding RNAs (ncRNAs) in the human genome have been reported to participate in the replication process of the influenza virus, among which there are still many unknowns about Long Intergenic Non-Coding RNAs (LincRNAs) in the cell cycle of viral infections. Here, we observed an increased expression of Linc01615 in A549 cells upon influenza virus PR8 infection, accompanied by the successful activation of the intracellular immune system. The knockdown of Linc01615 using the shRNAs promoted the proliferation of the influenza A virus, and the intracellular immune system was inhibited, in which the expressions of IFN-β, IL-28A, IL-29, ISG-15, MX1, and MX2 were decreased. Predictions from the catRAPID website suggested a potential interaction between Linc01615 and DHX9. Also, knocking down Linc01615 promoted influenza virus proliferation. The subsequent transcriptome sequencing results indicated a decrease in Linc01615 expression after influenza virus infection when DHX9 was knocked down. Further analysis through cross-linking immunoprecipitation and high-throughput sequencing (CLIP-seq) in HEK293 cells stably expressing DHX9 confirmed the interaction between DHX9 and Linc01615. We speculate that DHX9 may interact with Linc01615 to partake in influenza virus replication and that Linc01615 helps to activate the intracellular immune system. These findings suggest a deeper connection between DHX9 and Linc01615, which highlights the significant role of Linc01615 in the influenza virus replication process. This research provides valuable insights into understanding influenza virus replication and offers new targets for preventing influenza virus infections.

Computational design, docking, and molecular dynamics simulation study of RNA helicase inhibitors of dengue virus.

RNA viruses are complex pathogens in terms of their genetic makeup, mutation frequency, and transmission modes. They contain the RNA helicase enzyme, which plays a crucial role in the viral genome replication process. This work aims to develop and screen a potential molecule that could function as a dengue virus (DENV) RNA helicase inhibitor. The present study was performed by taking 26 potential derivatives of gedunin phytochemicals from the PubChem database as ligands. The binding study of the compounds were analyzed by in silico docking method considering DENV RNA helicase enzyme as the receptor. After a thorough analysis of the docking scores, toxicity, and physicochemical properties, compound tetrahydrogedunin was obtained as the best. Based on tetrahydrogedunin molecular structure, 100 drug-like molecules were designed using the Data Warrior tool. After the screening process for drug-likeness and ADMET properties, the derivative number 42 was considered as the promising. Further comparative docking of derivative 42 and a standard inhibitor molecule ST-610 with DENV RNA helicase enzyme showed binding affinity as -10.0 kcal/mol and -9.6 kcal/mol, respectively. The favorable interaction between DENV RNA helicase and derivative 42 was further validated by 50 nanoseconds molecular dynamics simulation and MM-GBSA analysis. Since the antiviral activity of derivative 42 has not been reported till date, the compound was predicted as a novel therapeutic molecule that can act against the dengue virus (DENV) RNA helicase enzyme.