Tariq Syeda Sumayya, Sardar Madiha, Shafiq Muhammad, Heinz Hendrick, Nur-E-Alam Mohammad, Ahmed Aftab, Ul-Haq Zaheer
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan.
H.E.J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan.
PLoS One. 2025 Jun 2;20(6):e0320865. doi: 10.1371/journal.pone.0320865. eCollection 2025.
Ecdysterone, often dubbed a "natural steroid," has garnered significant attention among athletes for its reputed growth-promoting and anabolic properties. Unlike synthetic anabolic steroids, which are classified as controlled substances, ecdysteroids remain largely unregulated in many countries and are widely marketed as dietary supplements. Notably, ecdysterone has been included in the World Anti-Doping Agency (WADA) monitoring program, highlighting its potential impact on athletic performance and raising questions about its regulation. Emerging evidence indicates that, unlike traditional anabolic steroids that act primarily via the Androgen Receptor (AR), ecdysterone's anabolic effects may be mediated through Estrogen Receptors (ERs), particularly Estrogen Receptor beta (ERβ). Despite these insights, the precise molecular mechanisms underlying ecdysterone's biological activity remain poorly characterized, particularly from an in-silico perspective. This paper aims to address these gaps by exploring ecdysterone's mechanism of action through computational and molecular modeling approaches. This study employs an advanced computational framework to unravel the binding dynamics and interaction mechanisms of ecdysterone with Androgen Receptor (AR), Estrogen Receptor alpha (ERα), and Estrogen Receptor beta (ERβ). Using chemical descriptor analysis, inter-molecular interaction mapping, and all-atom molecular dynamics simulations spanning 250 ns for each system, the study reveals that ecdysterone preferentially binds to ERβ, forming stable and compact complexes characterized by minimal per-residue fluctuations as evident in the average RMSD, RMSF, and Rg values observed for ERβ - Ecdysterone as 1.98 ± 0.31 Å, 1.07 ± 0.52 Å, and 18.44 ± 0.08 Å respectively which are significantly comparable with the ERβ - native complex, while high hydrogen bond occupancy was also observed for ERβ - Ecdysterone complex. Although binding free energy calculations suggest stronger interactions with ERα, the associated high fluctuations diminish its binding efficacy. In contrast, interactions with ERβ remain consistent and robust. Machine learning-based principal component analysis highlights coordinated motion patterns, while free energy profiles demonstrate stable energy basins with minimal variation. These findings underscore the pivotal role of ERβ in mediating ecdysterone's anabolic effects, distinguishing it from traditional androgenic steroids, and provide critical insights into its unique mechanism of action. This work lays the foundation for further exploration of ecdysterone as a potential anabolic agent.
蜕皮甾酮，常被称为“天然类固醇”，因其所谓的促进生长和合成代谢特性而在运动员中备受关注。与被归类为管制物质的合成合成代谢类固醇不同，蜕皮甾类在许多国家基本不受监管，并作为膳食补充剂广泛销售。值得注意的是，蜕皮甾酮已被纳入世界反兴奋剂机构（WADA）的监测计划，这凸显了其对运动成绩的潜在影响，并引发了对其监管的质疑。新出现的证据表明，与主要通过雄激素受体（AR）起作用的传统合成代谢类固醇不同，蜕皮甾酮的合成代谢作用可能通过雌激素受体（ERs）介导，特别是雌激素受体β（ERβ）。尽管有这些见解，但蜕皮甾酮生物活性的精确分子机制仍不清楚，尤其是从计算机模拟的角度来看。本文旨在通过计算和分子建模方法探索蜕皮甾酮的作用机制，以填补这些空白。本研究采用先进的计算框架来揭示蜕皮甾酮与雄激素受体（AR）、雌激素受体α（ERα）和雌激素受体β（ERβ）的结合动力学和相互作用机制。通过化学描述符分析、分子间相互作用图谱以及对每个系统进行250纳秒的全原子分子动力学模拟，研究发现蜕皮甾酮优先与ERβ结合，形成稳定而紧密的复合物，其特征是每个残基的波动最小，这在ERβ - 蜕皮甾酮的平均RMSD、RMSF和Rg值中很明显，分别为1.98±0.31 Å、1.07±0.52 Å和18.44±0.08 Å，与ERβ - 天然复合物具有显著可比性，同时在ERβ - 蜕皮甾酮复合物中也观察到高氢键占有率。尽管结合自由能计算表明与ERα的相互作用更强，但相关的高波动降低了其结合效率。相比之下，与ERβ的相互作用保持一致且稳健。基于机器学习的主成分分析突出了协同运动模式，而自由能分布表明存在稳定的能量盆地，变化最小。这些发现强调了ERβ在介导蜕皮甾酮合成代谢作用中的关键作用，使其有别于传统的雄激素类固醇，并为其独特的作用机制提供了重要见解。这项工作为进一步探索蜕皮甾酮作为潜在的合成代谢剂奠定了基础。
