Lu Binbin, Zhang Qian, Lin Hang, Wang Shaowei, Zhu Yang, Ge Ren-Shan, Li Xingwang
Department of Anesthesiology and Perioperative Medicine, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325027, China; Key Laboratory of Pediatric Anesthesiology, Ministry of Education, Wenzhou Medical University, Wenzhou, Zhejiang 325027, China; Key Laboratory of Precision Anesthesiology of Zhejiang Province, Wenzhou Medical University, Wenzhou, Zhejiang 325027, China.
Department of Obstetrics and Gynecology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China.
J Steroid Biochem Mol Biol. 2025 Oct;253:106794. doi: 10.1016/j.jsbmb.2025.106794. Epub 2025 May 31.
We investigated the inhibitory effects of dithiocarbamates (DTCs) on human steroid 5α-reductase 1 (SRD5A1), an enzyme crucial for the conversion of testosterone or pregnenolone into neuroactive steroids. Utilizing a comprehensive approach that included enzyme assays, molecular docking simulations, and both structure-activity relationship (SAR) and 3D quantitative SAR (3D-QSAR) analyses, we assessed the potency and interaction mechanisms of DTCs with SRD5A1 in human SF126 cells. Our results demonstrated that zinc dibutyldithiocarbamate, disulfiram, ferbam, thiram, and ziram displayed significant inhibitory activity against SRD5A1, with IC values of 1.10, 1.62, 2.31, 1.74, and 1.84 μM, respectively. Kinetic studies indicated that these compounds function as mixed inhibitors, suggesting a multifaceted interaction with the enzyme's active site. These DTCs effectively penetrated the cell membrane of SF126 cells to inhibit SRD5A1 at ≥ 5 μM. Molecular docking analyses revealed that these compounds interacted with a distinct domain located between the NADPH and testosterone binding sites of SRD5A1, primarily through sulfur bonds, which are a fundamental component of the DTC structure. Experimental validation of the inhibitory mechanism was achieved by demonstrating that the co-incubation of dithiothreitol, a sulfhydryl-reducing agent, significantly reversed the inhibitory effects of zinc dibutyldithiocarbamate and thiram. SAR analysis revealed a negative correlation between LogP, molecular weight, and IC values, and a positive correlation between LogS, lowest binding energy, and IC values. The 3D-QSAR analysis further indicated that hydrogen bond acceptor and hydrophobic region capabilities significantly contribute to the primary inhibition. In conclusion, this research significantly advances our understanding of the SAR of sulfur-containing DTCs as inhibitors of human SRD5A1, providing valuable insights for identifying toxicity of DTCs targeting this enzyme.
我们研究了二硫代氨基甲酸盐（DTCs）对人甾体5α-还原酶1（SRD5A1）的抑制作用，该酶对于睾酮或孕烯醇酮转化为神经活性甾体至关重要。我们采用了包括酶活性测定、分子对接模拟以及结构-活性关系（SAR）和三维定量构效关系（3D-QSAR）分析在内的综合方法，评估了DTCs在人SF126细胞中与SRD5A1的效力和相互作用机制。我们的结果表明，二丁基二硫代氨基甲酸锌、双硫仑、福美铁、福美双和福美锌对SRD5A1表现出显著的抑制活性，其IC值分别为1.10、1.62、2.31、1.74和1.84 μM。动力学研究表明，这些化合物起混合抑制剂的作用，表明它们与酶的活性位点存在多方面的相互作用。这些DTCs在浓度≥5 μM时能有效穿透SF126细胞的细胞膜以抑制SRD5A1。分子对接分析显示，这些化合物与位于SRD5A1的NADPH和睾酮结合位点之间的一个独特结构域相互作用，主要通过硫键，而硫键是DTC结构的基本组成部分。通过证明巯基还原剂二硫苏糖醇的共孵育显著逆转了二丁基二硫代氨基甲酸锌和福美双的抑制作用，实现了对抑制机制的实验验证。SAR分析显示LogP、分子量与IC值之间呈负相关，而LogS、最低结合能与IC值之间呈正相关。3D-QSAR分析进一步表明，氢键受体和疏水区域能力对主要抑制作用有显著贡献。总之，本研究极大地推进了我们对含硫DTCs作为人SRD5A1抑制剂的构效关系的理解，为识别靶向该酶的DTCs的毒性提供了有价值的见解。
