三苯基膦柱[5]芳烃的制备及其对金黄色葡萄球菌的抑制

Synthesis of triphenylphosphine pillar[5]arene with inhibitory effect on Staphylococcus aureus

摘要:

金黄色葡萄球菌是引起感染的最常见病原菌之一，较易黏附在介质或植入物表面形成生物被膜，使其较难清除和产生抗生素耐药性，给临床治疗带来极大挑战。因此研发新型的抑菌剂是非常急需的。柱[5]芳烃是一类新型的超分子大环宿主，因其高度刚性和对称的结构，以及可控的腔体尺寸，为创造具有不同官能团和生物活性的各种抗菌剂提供了无限的可能性。【目的】 合成三苯基膦柱[5]芳烃(triphenylphosphine pillar[5]arene, TPP)，以金黄色葡萄球菌(Staphylococcus aureus) ATCC 6538、金黄色葡萄球菌亚种(Staphylococcus aureus subsp. aureus, S. subsp. aureus) ATCC 29213和耐甲氧西林金黄色葡萄球菌(methicillin-resistant S. aureus, MRSA) ATCC 43300为供试菌，进行抑菌性能和耐药性的探究。【方法】 通过最小抑菌浓度(minimal inhibitory concentration, MIC)和最小杀菌浓度(minimal bacteriocidal concentration, MBC)的确定，来评估化合物的抑菌活性。进一步通过结晶紫染色定量分析TPP对细菌生物被膜的形成及采用苯酚硫酸法测定生物被膜中胞外多糖的含量，最后进行亚抑制浓度传代培养对TPP的耐药性进行了研究。【结果】 TPP对 3种菌株均表现出较好的抑菌和杀菌活性，其对3种菌株的MIC均为15.63μg/mL；对S. aureus和S. subsp. aureus的MBC都为125.00μg/mL，而TPP即使在125.00 μg/mL浓度下也不能杀死MRSA。另外，TPP在MIC下对3种菌株的生物被膜抑制率分别为72.9%、69.2%和71.8%；生物被膜中胞外多糖含量随着TPP浓度的增加呈现下降的趋势；抗性发展实验结果表明，TPP对金黄色葡萄球菌在20次传代培养后均未产生耐药性。【结论】 本研究结果明确了TPP的抑菌性能，为其在新型抑菌剂开发领域提供了一定的理论依据。

Abstract:

Staphylococcus aureus is one of the common pathogens causing infections. It can attach media or implant surfaces to form biofilms, which makes it difficult to be tackled and leads to the generation of drug resistance, posing a great challenge to clinical treatment. Therefore, it is urgent to develop novel antimicrobials. Pillar[5]arenes, a new class of supramolecular macrocyclic hosts, attracting wide attention due to their highly rigid and symmetrical architectures and controllable cavity sizes, which afford the limitless possibility to create antimicrobial agents with various functional groups and biological activities. [Objective] To synthesize triphenylphosphine pillar[5]arene (TPP) and determine its antibacterial activities and drug resistance with Staphylococcus aureus ATCC 6538, Staphylococcus aureus subsp. aureus (S. subsp. aureus) ATCC 29213, and methicillin-resistant S. aureus ATCC 43300. [Methods] The minimal inhibitory concentration (MIC) and minimal bacteriocidal concentration (MBC) were determined to evaluate the antibacterial activity of TPP. The effects of TPP on biofilm formation were quantified by crystal violet staining, and the content of extracellular polysaccharides in the biofilm was determined by the phenol-sulfuric acid assay. the strain resistance to TPP was examined. [Results] TPP exhibited inhibitory effects on the three strains tested, with a MIC of 15.63 μg/mL for the three strains and a MBC of 125.00 μg/mL for both S. aureus and S. subsp. aureus. However, TPP was unable to kill MRSA even at a concentration of 125.00 μg/mL. The biofilm inhibition rates of TPP at MIC were as high as 72.9%, 69.2%, and 71.8% for the three strains, respectively. The content of extracellular polysaccharides decreased with the increase in the concentration of TPP. S. aureus did not develop resistance to TPP after 20 generations. [Conclusion] This study clarified the antibacterial performance of TPP, providing a theoretical basis for the further development and utilization of TPP in the medicine field.