Pyrancoumarin derivative LP4C targeting of pyrimidine de novo synthesis pathway inhibits MRSA biofilm and virulence

Liu, Yongsheng; Su, Shan; Yu, Moxi; Zhai, Dongshen; Hou, Yachen; Zhao, Hui; Ma, Xue; Jia, Min; Xue, Xiaoyan; Li, Mingkai

doi:10.3389/fphar.2022.959736

Cited by 6 publications

(5 citation statements)

References 41 publications

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“…It is grouped in α-type (PSMα and PSMγ) and β-type (PSMβ) peptides ( 37 , 38 ). Research has shown that pyrancoumarin derivative LP4C effectively attenuates MRSA biofilm development by downregulating the expression of the genes psmα and psmβ ( 39 ). In this study, we found that the genes psmα and psmβ were also notably reduced after the treatment of S-342-3.…”

Section: Discussionmentioning

confidence: 99%

A novel small-molecule compound S-342-3 effectively inhibits the biofilm formation of Staphylococcus aureus

Zhang,

Shen,

Zhou

et al. 2023

Microbiol Spectr

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Staphylococcus aureus is an important human pathogen in both community and hospital settings that often causes persistent and recurrent infections. The continuous emergence of multidrug-resistant strains and the lag in antibiotic development make the treatment of S. aureus more challenging. The biofilm formation of S. aureus is an important reason for persistent infection. In this study, we demonstrated that a small-molecule compound S-342-3 can effectively inhibit the biofilm formation of S. aureus . The crystal violet semi-quantitative assays revealed that at a sub-minimum inhibitory concentration of 4 µg/mL, S-342-3 reduced S. aureus biofilm mass by 57.43%, 52.14%, and 25.49%. Using confocal laser scanning microscopy, we observed that the biofilm was more incompact and less uniform upon the treatment of S-342-3. At a sub-inhibitory concentration (4 µg/mL), the S-342-3 can effectively reduce the production of polysaccharide intercellular adhesin (PIA) and suppress the initial adhesion of cells in the biofilm. Consistently, the results of RT-qPCR revealed that the expression of genes associated with biofilm formation was downregulated by S-342-3 ( P < 0.05). However, we found that treatment with S-342-3 resulted in a significant decrease in the expression of global regulatory genes agrA and sarA ( P < 0.05), which play a key role in promoting cell surface attachment and PIA production in S. aureus biofilms. Also importantly, we experimentally proved that the S-342-3 was not toxic to A549 alveolar epithelial cells and the Galleria mellonella larvae. Collectively, these results suggest that the S-342-3 may be a promising anti-biofilm drug candidate with excellent biosafety, which can be further explored for its practical application in health care. IMPORTANCE Biofilms are an important virulence factor in Staphylococcus aureus and are characterized by a structured microbial community consisting of bacterial cells and a secreted extracellular polymeric matrix. Inhibition of biofilm formation is an effective measure to control S. aureus infection. Here, we have synthesized a small molecule compound S-342-3, which exhibits potent inhibition of biofilm formation in both MRSA and MSSA. Further investigations revealed that S-342-3 exerts inhibitory effects on biofilm formation by reducing the production of polysaccharide intercellular adhesin and preventing bacterial adhesion. Our study has confirmed that the inhibitory effect of S-342-3 on biofilm is achieved by downregulating the expression of genes responsible for biofilm formation. In addition, S-342-3 is non-toxic to Galleria mellonella larvae and A549 cells. Consequently, this study demonstrates the efficacy of a biologically safe compound S-342-3 in inhibiting biofilm formation in S. aureus, thereby providing a promising antibiofilm agent for further research.

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Section: Discussionmentioning

confidence: 99%

A novel small-molecule compound S-342-3 effectively inhibits the biofilm formation of Staphylococcus aureus

Zhang,

Shen,

Zhou

et al. 2023

Microbiol Spectr

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“…Our study developed the widely applicable and efficient PyrR-based CRISPR-Cas system in L. casei for whole genetic deletion, which provided novel insights into the molecular mechanisms of the PyrR gene in relation to the antimicrobial antagonism of metabolic products, competition with pathogens for adhesion, and immunoregulation. The molecular docking results indicated that the pyrR protein attenuates the virulence of biofilm formation and targets the de novo pyrimidine synthesis pathway in S. aureus [14]. To increase the sensitivity of 5-FOA, the double-mutant strain ∆pyrF ∆pyrR in Geobacillus kaustophilus was constructed.…”

Section: Discussionmentioning

confidence: 99%

Loss in the Antibacterial Ability of a PyrR Gene Regulating Pyrimidine Biosynthesis after Using CRISPR/Cas9-Mediated Knockout for Metabolic Engineering in Lactobacillus casei

Chen,

He,

Qin

et al. 2023

Microorganisms

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Lactobacillus casei (L. casei) has four possible mechanisms: antimicrobial antagonism, competitional adhesion, immunoregulation, and the inhibition of bacterial toxins. To delineate the metabolic reactions of nucleotides from L. casei that are associated with mechanisms of inhibiting pathogens and immunoregulation, we report that a PyrR-deficient L. casei strain was constructed using the CRISPR-Cas9D10A tool. Furthermore, there were some changes in its basic biological characterization, such as its growth curve, auxotroph, and morphological damage. The metabolic profiles of the supernatant between the PyrR-deficient and wild strains revealed the regulation of the synthesis of genetic material and of certain targeting pathways and metabolites. In addition, the characteristics of the PyrR-deficient strain were significantly altered as it lost the ability to inhibit the growth of pathogens. Moreover, we identified PyrR-regulating pyrimidine biosynthesis, which further improved its internalization and colocalization with macrophages. Evidence shows that the PyrR gene is a key active component in L. casei supernatants for the regulation of pyrimidine biosynthesis against a wide range of pathogens.

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“…Coumarins are important phenolic substances and exhibit the broad-spectrum anti-biofilm activity against a wide range of bacteria [ 21 , 22 , 23 ]. Our previous results showed that coumarin derivative LP4C was the potential anti-bacterial biofilm compound in vitro and in vivo [ 18 , 19 ]. The present study further confirmed that LP4C inhibited bacterial biofilm formation even at 5 μg/mL, which was about 80 times lower than the other reported coumarin derivatives, such as esculetin (6,7-dihydroxycoumarin) and esculin [ 24 ].…”

Section: Discussionmentioning

confidence: 99%

“…Our previous study showed that 4-hydroxycoumarin derivate DCH could inhibit the biofilm formation of methicillin-resistant Staphylococcus aureus (MRSA) selectively [ 17 ]. Interestingly, we screened another coumarin derivate LP4C which exhibited significant inhibitory activity to biofilm formation in S. aureus and P. aeruginosa [ 18 , 19 ]. These results highlight the potential role of coumarin derivatives as new therapeutic agents to combat the bacterial biofilm infection.…”

Section: Introductionmentioning

confidence: 99%

Pharmacological Characters and Toxicity Evaluation of Coumarin Derivative LP4C as Lead Compound against Biofilm Formation of Pseudomonas aeruginosa

Xin

Liu

et al. 2023

Molecules

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Pseudomonas aeruginosa-induced biofilm infection is difficult to treat and poses a significant threat to public health. Our previous study found a new coumarin derivative LP4C which exerted potent in vitro and in vivo anti-biofilm activity against Pseudomonas aeruginosa; however, the underlying molecular mechanism and drug-likeness of LP4C is unclear. In this study, we confirmed that LP4C could inhibit the biofilm in dose-dependent manner without bactericidal activity. The transcriptomic profiling and RT-PCR result revealed that bacterial pyrimidine mediated the inhibitory activity of LP4C. The cell viability was not affected in LP4C treatment groups with the concentration under 200 μg/mL, and no death or toxicity sign was observed in mice treated by 20, 40 and 80 mg/kg LP4C during the three-week test period. Ames test presented that LP4C had no effect on the bacterial reverse mutation. In additional, pharmacokinetic results showed that LP4C was likely to have the orally bioavailable properties. Our data indicate that LP4C is a possible lead compound for the development of new anti-biofilm infection agents against Pseudomonas aeruginosa.

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Pyrancoumarin derivative LP4C targeting of pyrimidine de novo synthesis pathway inhibits MRSA biofilm and virulence

Cited by 6 publications

References 41 publications

A novel small-molecule compound S-342-3 effectively inhibits the biofilm formation of Staphylococcus aureus

A novel small-molecule compound S-342-3 effectively inhibits the biofilm formation of Staphylococcus aureus

Loss in the Antibacterial Ability of a PyrR Gene Regulating Pyrimidine Biosynthesis after Using CRISPR/Cas9-Mediated Knockout for Metabolic Engineering in Lactobacillus casei

Pharmacological Characters and Toxicity Evaluation of Coumarin Derivative LP4C as Lead Compound against Biofilm Formation of Pseudomonas aeruginosa

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