Since our study showed that sulfone derivatives' action mode creates a lesser risk of inducing widespread resistance among Candida spp., we continued verifying sulfones' antifungal activity using the following newly synthesized derivatives: bromodichloromethy-4-hydrazinyl-3-nitrophenyl sulfone (S1), difluoroiodomethyl-4-hydrazinyl-3-nitrophenyl sulfone (S2), and chlorodifluoromethyl-4-hydrazinyl-3-nitrophenyl sulfone (S3). As the mechanism by which sulfones gain access to the cytoplasm has not been elucidated yet, in order to track S1-3, we coupled their hydrazine group with BODIPY (final S1-3 BODIPY-labelled were named SB1-3). This approach allowed us to follow the vital internalization and endocytic routing of SB1-3, while BODIPY interacts primarily with fungal surfaces, thus confirming that S1-3 and their counterparts SB1-2 behaved as non-typical agents by damaging the cell membrane and wall after being endocytosed (SB1-3 fluorescence visible inside the unlysed sessile cells). Thus greatly decreasing the likelihood of the appearance of strains resistance. Core sulfones S1-3 are a promising alternative not only to treat planktonic C. albicans but also biofilm-embedded cells. In the flow cytometric analysis, the planktonic cell surface was digested by S1-3, which made the externalized PS accessible to AnnexinV binding and PI input (accidental cell death ACD). The occurrence of ACD as well as apoptosis (crescent-shaped nuclei) and anoikis of sessile cells (regulated cell death by 100%-reduction in attachment to epithelium) was assessed through monitoring the AO/PI/HO342 markers. CLSM revealed the invasion of S1-3 and SB1-3 in C. albicans without inducing cell lysis. This was a novel approach in which QCM-D was used for real-time in situ detection of viscoelastic changes in the C. albicans biofilm, and its interaction with S1 as a representative of the sulfones tested. S1 (not toxic in vivo) is a potent fungicidal agent against C. albicans and could be administered to treat invasive candidiasis as a monotherapy or in combination with antifungal agents of reference to treat C. albicans infections.

中文翻译：

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砜衍生物进入白色念珠菌无柄细胞的细胞质。

由于我们的研究表明砜衍生物的作用方式在念珠菌属中引起广泛抗药性的风险较小，因此我们继续使用以下新合成的衍生物验证砜类的抗真菌活性：溴二氯甲基-4-肼基-3-硝基苯砜（S1） ，二氟碘甲基-4-肼基-3-硝基苯基砜（S2）和氯二氟甲基-4-肼基-3-硝基苯基砜（S3）。由于尚未阐明砜进入细胞质的机制，为了追踪S1-3，我们将其肼基与BODIPY偶联（最终的S1-3 BODIPY标记为SB1-3）。这种方法使我们能够遵循SB1-3的重要内在化和内吞途径，而BODIPY主要与真菌表面相互作用，因此证实了S1-3及其对应的SB1-2在被内吞后通过破坏细胞膜和壁（未溶解的无柄细胞内可见的SB1-3荧光）而充当了非典型药物。因此大大降低了出现应变抗性的可能性。核心砜S1-3是一种有前途的替代品，不仅可以治疗浮游白色念珠菌，而且还可以用于生物膜包埋的细胞。在流式细胞仪分析中，浮游生物的细胞表面被S1-3消化，这使得膜联蛋白V结合和PI输入（意外细胞死亡ACD）可接近外部PS。通过监测AO / PI / HO342标记来评估ACD的发生以及无细胞凋亡（新月形核）和无神经（通过上皮附着的100％减少来调节细胞死亡）。CLSM揭示了白色念珠菌S1-3和SB1-3的入侵而没有诱导细胞溶解。这是一种新颖的方法，其中QCM-D用于实时原位检测白色念珠菌生物膜中的粘弹性变化及其与S1的相互作用（作为所测试的砜的代表）。S1（在体内无毒）是一种有效的抗白色念珠菌杀真菌剂，可作为单一疗法或与参考抗真菌剂联合使用，以治疗侵入性念珠菌病，以治疗白色念珠菌感染。