Identification, prioritization, and evaluation of RlpA protein as a target against multidrug-resistant Pseudomonas aeruginosa

• Published in: Acta tropica Vol. 255; p. 107216
• Main Authors: Gatasheh, Mansour K., Murugan, Nandagopal, Krishnamoorthy, Rajapandiyan, Alshuniaber, Mohammad A., Malathi, Jambulingam, Umashankar, Vetrivel, Ramalingam, Gopinath, Veeraraghavan, Vishnu Priya, Jayaraman, Selvaraj
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.07.2024
• Subjects: Anti-pseudomonas activity; Drug target; Infectious disease; Molecular docking; Multi-drug resistant; Virtual screening; Drug target; Virtual screening; Infectious disease; Multi-drug resistant; Anti-pseudomonas activity; Molecular docking
• ISSN: 0001-706X; 1873-6254; 1873-6254
• DOI: 10.1016/j.actatropica.2024.107216

•Septicemia exhibits a higher morbidity and mortality rate compared to infections caused by susceptible Pseudomonas aeruginosa (P. aeruginosa). In the case of ocular infections mediated by multidrug-resistant (MDR) P. aeruginosa, there is a potential for significant vision impairment or blindness.•This research utilized advanced genomics-based techniques to identify potential drug targets within P. aeruginosa, with a specific focus on the bacterial septal ring protein RlpA.•The target protein chosen was Rare Lipo Protein A (RlpA), and its crystal structure underwent geometric optimization using NCBI data collection, excluding human non-homologs, and applying other relevant criteria.•Virtual screening and docking techniques were applied, followed by contacting the manufacturer to acquire the shortlisted small molecules. The purchased compounds were then tested against drug-susceptible, MDR, and pan-drug-resistant P. aeruginosa using Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) tests.•Among the tested compounds, 3-Chlorophthalic acid displayed significant anti-Pseudomonas activity. Subsequent validation included assessing its toxicity levels through the MTT assay and confirming its specificity against the RlpA gene via reverse transcriptase-real-time PCR. The study demonstrated heightened anti-Pseudomonas activity coupled with reduced toxicity to host cells.•These findings hold promise for potential clinical studies, suggesting that 3-Chlorophthalic acid could provide a viable solution to combat infections caused by multidrug-resistant P. aeruginosa, addressing a critical challenge in healthcare. According to the World Health Organization, infectious diseases, particularly those caused by multidrug-resistant bacteria (MDR), are projected to claim the lives of 15 million people by 2050. Septicemia carries a higher morbidity and mortality rate than infections caused by susceptible Pseudomonas aeruginosa, and MDR-mediated ocular infections can lead to impaired vision and blindness. To identify and develop a potential drug against MDR P. aeruginosa, we employed in silico reverse genetics-based target mining, drug prioritization, and evaluation. Rare Lipoprotein A (RlpA) was selected as the target protein, and its crystal structure was geometrically optimized. Molecular docking and virtual screening analyses revealed that RlpA exhibits strong binding affinity with 11 compounds. Among these, 3-chlorophthalic acid was evaluated, and subsequent in vitro assays demonstrated significant anti-Pseudomonas activity with negligible cytotoxicity. The compound was further evaluated against both drug-susceptible and MDR P. aeruginosa strains in vitro, with cytotoxicity assessed using an MTT assay. The study demonstrated that 3-chlorophthalic acid exhibits potent anti-Pseudomonas activity with minimal toxicity to host cells. Consequently, this compound emerges as a promising candidate against MDR P. aeruginosa, warranting further investigation.