Hilpert Kai, Rumancev Christoph, Gani Jurnorain, Collis Dominic W P, Lopez-Perez Paula Matilde, Garamus Vasil M, Mikut Ralf, Rosenhahn Axel
Institute of Infection and Immunology, St. George's, University of London (SGUL), London, United Kingdom.
Laboratory Analytical Chemistry-Biointerfaces, Ruhr-University Bochum, Bochum, Germany.
Front Pharmacol. 2023 Jul 18;14:1141785. doi: 10.3389/fphar.2023.1141785. eCollection 2023.
The opportunistic yeast is the most common cause of candidiasis. With only four classes of antifungal drugs on the market, resistance is becoming a problem in the treatment of fungal infections, especially in immunocompromised patients. The development of novel antifungal drugs with different modes of action is urgent. In 2016, we developed a groundbreaking new medium-throughput method to distinguish the effects of antibacterial agents. Using small-angle X-ray scattering for biological samples (BioSAXS), it is now possible to screen hundreds of new antibacterial compounds and select those with the highest probability for a novel mode of action. However, yeast (eukaryotic) cells are highly structured compared to bacteria. The fundamental question to answer was if the ultrastructural changes induced by the action of an antifungal drug can be detected even when most structures in the cell stay unchanged. In this exploratory work, BioSAXS was used to measure the ultrastructural changes of that were directly or indirectly induced by antifungal compounds. For this, the well-characterized antifungal drug Flucytosine was used. BioSAXS measurements were performed on the synchrotron P12 BioSAXS beamline, EMBL (DESY, Hamburg) on treated and untreated yeast . BioSAXS curves were analysed using principal component analysis (PCA). The PCA showed that Flucytosine-treated and untreated yeast were separated. Based on that success further measurements were performed on five antifungal peptides {1. Cecropin A-melittin hybrid [CA (1-7) M (2-9)], KWKLFKKIGAVLKVL; 2. Lasioglossin LL-III, VNWKKILGKIIKVVK; 3. Mastoparan M, INLKAIAALAKKLL; 4. Bmkn2, FIGAIARLLSKIFGKR; and 5. optP7, KRRVRWIIW}. The ultrastructural changes of indicate that the peptides may have different modes of action compared to Flucytosine as well as to each other, except for the Cecropin A-melittin hybrid [CA (1-7) M (2-9)] and optP7, showing very similar effects on . This very first study demonstrates that BioSAXS shows promise to be used for antifungal drug development. However, this first study has limitations and further experiments are necessary to establish this application.
机会性酵母是念珠菌病最常见的病因。市面上只有四类抗真菌药物，耐药性正成为真菌感染治疗中的一个问题，尤其是在免疫功能低下的患者中。开发具有不同作用方式的新型抗真菌药物迫在眉睫。2016年，我们开发了一种开创性的新的中通量方法来区分抗菌剂的效果。利用生物样品小角X射线散射（BioSAXS），现在可以筛选数百种新的抗菌化合物，并选择那些具有新作用方式最高可能性的化合物。然而，与细菌相比，酵母（真核）细胞具有高度的结构。需要回答的基本问题是，即使细胞中的大多数结构保持不变，是否能够检测到抗真菌药物作用所诱导的超微结构变化。在这项探索性工作中，BioSAXS被用于测量抗真菌化合物直接或间接诱导的超微结构变化。为此，使用了特征明确的抗真菌药物氟胞嘧啶。在同步加速器P12 BioSAXS光束线（德国汉堡DESY的欧洲分子生物学实验室）上对处理过和未处理过的酵母进行了BioSAXS测量。使用主成分分析（PCA）对BioSAXS曲线进行了分析。PCA显示氟胞嘧啶处理过的酵母和未处理过的酵母被区分开。基于这一成功，对五种抗真菌肽进行了进一步测量{1. 天蚕素A-蜂毒肽杂合体[CA(1-7)M(2-9)]，KWKLFKKIGAVLKVL；2. 舌蜂素LL-III，VNWKKILGKIIKVVK；3. Mastoparan M，INLKAIAALAKKLL；4. Bmkn2，FIGAIARLLSKIFGKR；5. optP7，KRRVRWIIW}。酵母的超微结构变化表明，这些肽与氟胞嘧啶以及彼此相比可能具有不同的作用方式，除了天蚕素A-蜂毒肽杂合体[CA(1-7)M(2-9)]和optP7对酵母显示出非常相似的效果。这项首次研究表明BioSAXS有望用于抗真菌药物开发。然而，这项首次研究存在局限性，需要进一步的实验来确立这种应用。
