George Hanson ^1* Joseph Adams ¹ Daveson I B Kepgang ² Luke S Zondagh ³ Lewis Tem Bueh ⁴ Andy Asante ⁵ Soham A Shirolkar ⁶ Maureen Kisaakye ⁷ Hem Bondarwad ⁸ Olaitan I. Awe ^9*

• ¹ Department of Parasitology, Noguchi Memorial Institute for Medical Research (NMIMR), College of Health Sciences (CHS), University of Ghana, P.O. Box LG 581, Legon, Accra, Ghana, Accra, Ghana
• ² Department of Biochemistry, Faculty of Sciences, University of Douala, P.O. Box 24157, Douala, Cameroon, Douala, Cameroon
• ³ Pharmaceutical Chemistry, School of Pharmacy, University of Western Cape Town, Private Bag X17, Belville 7535, Cape Town, South Africa, Cape Town, South Africa
• ⁴ Department of Computer Engineering, Faculty of Engineering and Technology, University of Buea, P.O. Box 63, Buea, Cameroon, Buea, Cameroon
• ⁵ Department of Immunology, Noguchi Memorial Institute for Medical Research (NMIMR), College of Health Sciences (CHS), University of Ghana, P.O. Box LG 581, Legon, Accra, Ghana, Accra, Ghana
• ⁶ College of Engineering, University of South Florida, 4202 E Fowler Ave, Tampa, FL 33620; Florida, U.S.A., Florida, United States
• ⁷ Department of Immunology and Molecular Biology, College of Health Sciences, Makerere University, P.O. Box 7072, Kampala, Uganda, Kampala, Uganda
• ⁸ Department of Biotechnology and Bioinformatics, Deogiri College, Dr. Babasaheb Ambedkar Marathwada University, Chh. Sambhajinagar, India, Sambhajinagar, India
• ⁹ African Society for Bioinformatics and Computational Biology, Cape Town, South Africa, Cape Town, South Africa

Dengue fever continues to pose a global threat due to the widespread distribution of its vector mosquitoes, Aedes aegypti and Aedes albopictus. While the WHO-approved vaccine, Dengvaxia, and antiviral treatments like Balapiravir and Celgosivir are available, challenges such as drug resistance, reduced efficacy, and high treatment costs persist. This study aimed to identify novel potential inhibitors of the Dengue virus (DENV) using an integrative drug discovery approach encompassing machine learning and molecular docking techniques. Utilizing a dataset of 21,250 bioactive compounds from PubChem (AID: 651640), alongside a total of 1,444 descriptors generated using PaDEL, we trained machine learning models such as Support Vector Machine, Random Forest, k-nearest neighbors, Logistic Regression, and Gaussian Naïve Bayes to classify compounds. The top-performing model was used to predict active compounds, followed by molecular docking performed using AutoDock Vina. The detailed interactions, toxicity, stability, and conformational changes of selected compounds were assessed through protein-ligand interaction studies, molecular dynamics (MD) simulations, and binding free energy calculations.We implemented a robust three-dataset splitting strategy, employing the Logistic Regression algorithm, which achieved an accuracy of 94%. The model successfully predicted 18 known DENV inhibitors, with 11 identified as active, paving the way for further exploration of 2,683 new compounds from the ZINC and EANPDB databases. Subsequent molecular docking studies were performed on the NS2B/NS3 protease, an enzyme essential in viral replication. ZINC95485940, ZINC38628344, 2',4'-dihydroxychalcone and ZINC14441502 demonstrated a high binding affinity of -8.1, -8.5, -8.6 and -8.0 kcal/mol, respectively, exhibiting stable interactions with His51, Ser135, Leu128, Pro132, Ser131, Tyr161, and Asp75 within the active site, which are critical residues involved in inhibition. Molecular dynamics simulations coupled with MMPBSA further elucidated the stability, making it a promising candidate for drug development.Overall, this integrative approach, combining machine learning, molecular docking, and dynamics simulations, highlights the strength and utility of computational tools in drug discovery. It suggests a promising pathway for the rapid identification and development of novel antiviral drugs against DENV. These in-silico findings provide a strong foundation for future experimental validations and in-vitro studies aimed at fighting DENV.