Isswanich Kulpornsorn, Poungcho Pattara, Buaban Koonchira, Chuntakaruk Hathaichanok, Sanachai Kamonpan, Chansriniyom Chaisak, Rungrotmongkol Thanyada, De-Eknamkul Wanchai, Chamni Supakarn
Pharmaceutical Sciences and Technology Program, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand.
Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand.
ACS Omega. 2025 Aug 13;10(33):38211-38228. doi: 10.1021/acsomega.5c06242. eCollection 2025 Aug 26.
The development of steroid 5α-reductase inhibitors for the treatment of androgenetic alopecia remains challenging, primarily due to the unavailability of a crystal structure for human steroid 5α-reductase type-1 (SRD5A1) and the adverse effects associated with existing steroidal therapeutic agents such as finasteride and dutasteride. In this study, hydroxycinnamate derivatives were synthesized and evaluated for their inhibitory activity against SRD5A1 using an immortalized human keratinocyte cell-based assay coupled with nonradioactive high-performance thin-layer chromatography analysis. Three ferulate derivatives featuring 2',5'-, 3',4'-, and 3',5'-bis-(trifluoromethyl)-phenyl substituents (, , and ) demonstrated inhibitory activity with half-maximal inhibitory concentrations (IC) of 8.50, 10.06, and 8.05 μM, respectively, exhibiting low cytotoxicity and favorable predicted lipophilicity. Compound was chosen for further investigation to assess its effect on SRD5A1 protein expression. Treatment of HaCaT cells with compound at concentrations of 10-15 μM for 24 h significantly downregulated SRD5A1 protein expression, suggesting its inhibitory effect on both SRD5A1 activity and expression. An model identified key binding interactions between the AlphaFold-generated SRD5A1 catalytic pocket and the residues R98, M119, A120, E202, and F228, among which M119 is recognized as a crucial residue for the stereoselective and irreversible reduction catalyzed by SRD5A1. Structure-activity relationship analysis indicated that the 3-methoxy-4-hydroxy core facilitates hydrogen bonding with R98, whereas the cinnamoyl and bis-(trifluoromethyl)-phenyl moieties enhance hydrophobic and halogen interactions with key residues, improving binding affinity and molecular orientation. Thus, these bis-(trifluoromethyl)-substituted phenyl ferulate derivatives present a promising class of nonsteroidal inhibitors for SRD5A1, with potential applications in developing topical antiandrogenic agents for the treatment androgen-related conditions.
开发用于治疗雄激素性脱发的甾体5α-还原酶抑制剂仍然具有挑战性，主要原因是缺乏人1型甾体5α-还原酶（SRD5A1）的晶体结构，以及现有甾体治疗药物（如非那雄胺和度他雄胺）存在不良反应。在本研究中，合成了羟基肉桂酸酯衍生物，并使用基于永生化人角质形成细胞的测定法结合非放射性高效薄层色谱分析，评估了它们对SRD5A1的抑制活性。三种具有2',5'-、3',4'-和3',5'-双（三氟甲基）苯基取代基的阿魏酸衍生物（、和）表现出抑制活性，其半数最大抑制浓度（IC）分别为8.50、10.06和8.05 μM，具有低细胞毒性和良好的预测亲脂性。选择化合物进行进一步研究，以评估其对SRD5A1蛋白表达的影响。用10-15 μM浓度的化合物处理HaCaT细胞24小时，可显著下调SRD5A1蛋白表达，表明其对SRD5A1活性和表达均有抑制作用。一个模型确定了AlphaFold生成的SRD5A1催化口袋与残基R98、M119、A120、E202和F228之间的关键结合相互作用，其中M119被认为是SRD5A1催化立体选择性和不可逆还原的关键残基。构效关系分析表明，3-甲氧基-4-羟基核心促进与R98形成氢键，而肉桂酰基和双（三氟甲基）苯基部分增强与关键残基的疏水和卤素相互作用，提高结合亲和力和分子取向。因此，这些双（三氟甲基）取代的苯基阿魏酸酯衍生物是一类有前景的SRD5A1非甾体抑制剂，在开发用于治疗雄激素相关疾病的局部抗雄激素药物方面具有潜在应用。
