In silico and in vitro insights into the prediction and analysis of natural photosensitive compounds targeting Acinetobacter baumannii biofilm-associated protein

• The computational drug design methods exhibited Bap is a stable protein in A. baumannii . • Molecular doking analysis showed that Cur and Qct interacted with Bap with high affinity. • Combination of LED light and these natural photosensitizers significantly reduced the growth of A. baumannii biofilm. • aPDT using Cur and Qct downregulated the gene involved in the biofilm formation of A. baumannii. The spread of Acinetobacter baumannii strains has become a global concern due to its extensive antibiotic resistance and biofilm formation. To overcome it, new antimicrobial strategies have been needed. Among them, antimicrobial photodynamic therapy (aPDT) is an efficient approach against various microorganisms. This study was focused on the use of curcumin (Cur) and quercetin (Qct) as natural photosensitive compounds to improve the activity of aPDT against A. baumannii biofilm-associated protein (Bap). In this in silico and in vitro study, after determining drug-likeness property, ADME/Toxicity profile, and pharmacological activity of Cur and Qct, virtual screening and molecular docking were assessed to determine the potential binding modes of Cur and Qct to Bap. Then, the anti-biofilm potential of natural photosensitizers-mediated aPDT against A. baumannii was evaluated after the determination of minimum inhibitory concentration (MIC). Subsequently, reverse transcription-quantitative real-time PCR (RT-qPCR) was used to exhibit the anti-virulent effect of aPDT against the gene involved in the biofilm formation of A. baumannii Cur and Qct showed almost similar pharmacokinetic and pharmacodynamics properties. These natural photosensitizers obeyed all the criteria of Lipinski's rule of five principles. According to the molecular docking analysis of protein-ligand complexes, Qct and Cur with a high affinity for Bap showed binding affinity of -6.34 and -6.98 kcal/mol, respectively. According to the findings, aPDT using 4 ×, and 8 × MIC of Cur and Qct could significantly reduce A. baumannii growth in biofilm structures in comparison with the control group ( P < 0.05). Also, a significant downregulation by 3.7-, and 5.2-fold in gene expression of bap was observed after treatment with sub-MIC doses of Cur- and Qct-mediated aPDT, respectively ( P < 0.05). In summary, the in silico analysis showed that Cur and Qct had strong binding affinity with Bap as a stable protein of A. baumannii . Furthermore, in vitro results displayed that targeted aPDT based on these natural photosensitizers can be considered a treatment against A. baumannii infections by reducing the growth of microbial biofilm and reducing the expression of bap as a gene involved in A. baumannii biofilm formation.