Supplementary materials Nephthea sp. inhibits Biofilm, DNA gyrase, HSP90, and DHFR: In vitro, in silico, and pharmacokinetics studies Nevine H. Hassan a , Seham S. El-Hawary b , Mahmoud Emam c , Nesreen A. Safwat g , Mohamed A. Rabeh b,d , Usama Ramadan Abdelmohsen e,f* and Nabil M. Selim b* a) Pharmacognosy Department, Faculty of Pharmacy, Modern University for Technology and Information, Cairo 11571, Egypt b) Pharmacognosy Department, Faculty of Pharmacy, Cairo University, Giza 11562, Egypt c) Phytochemistry and Plant Systematics Department, National Research Centre, Dokki, Cairo 12622, Egypt (M.E.) d) Pharmacognosy Department, College of Pharmacy, King Khalid University, Abha 61441, Saudi Arabia e) Pharmacognosy Department, Faculty of Pharmacy, Minia University, 61519 Minia, Egypt f) Pharmacognosy Department, Faculty of Pharmacy, Deraya University, 61111 New Minia, Egypt g) Microbiology and Immunology Department, Faculty of Pharmacy, Modern University for Technology and Information, Cairo 11571, Egypt * Equally contributed and Corresponding authors: Prof. Dr. Usama Ramadan Abdelmohsen (e-mail: Usama.ramadan@mu.edu.eg, Tel.: +2-86-2347759, Fax: +2-86-2369075) , and Assoc. Prof. Dr. Nabil M. Selim (e-mail: nabil.selim@pharma.cu.edu.eg, Tel.:+201117185050, Fax: +02-32628426 ) Abstract This study attempts to identify and assess a novel marine-derived antibiofilm agent. The antibacterial activity of n-hexane, dichloromethane, ethyl acetate, and butanol fractions from the crude extract of soft coral Nephthea sp. was evaluated against six microorganisms. As the ethyl acetate fraction was most effective against Bacillus subtilis, Escherichia coli, and Candida, investigated potential biofilm inhibition against the tested strains. Seventeen secondary metabolites were identified using (UPLC-Q/TOF-MS) responsible for these biological activities of the active fraction. Additionally, a molecular docking study was done and showed free binding energy of -7.5 kcal/mol; Azamial A had the highest binding affinity for the DNA gyrase enzyme, while Sinularectin had -8.3 and -7.6 kcal/mol for the DHFR and HSP90 enzymes, respectively. Moreover, pharmacokinetics and (ADME) studies for Azamia9l A and Sinularectin