The conversion of 17-oxosteroids to 17β-hydroxysteroids stands as a pivotal process in the synthesis of numerous steroidal drugs and intermediates. This study explored the potential of the strain Priestia aryabhattai (IICT-BC-1279) to catalyze the reduction of the C-17 carbonyl group in androst-4-ene-3,17-dione (AD), resulting in the exclusive production of testosterone (TS) through its 17β-hydroxysteroid dehydrogenase (17β-HSD) activity. Optimal conditions for this reduction were achieved at pH 7.0 and 25 °C, with supplementation of AD as an inducer (0.01 g/L), 1 % Tween 80 (w/v) and ethanol as co-solvent. Under these optimized parameters, 0.5 g/L AD was efficiently converted to TS as a sole product, achieving a yield of > 95 % and diastereomeric excess (d.e) of > 99 % within 48 h. The absence of by-products in this microbial 17β-reduction process simplifies product purification, highlighting the strain’s potential as a valuable biocatalyst for this essential transformation. Additionally, the conversion of androsta-1,4-diene-3,17-dione (ADD) to (+)-Boldenone (BD) was studied to explore substrate scope.
{"title":"Microbial steroid biotransformation: Regio- and stereo selective 17β-reduction by Priestia aryabhattai","authors":"Yogeswari Sudhakar , Reddy Krishna Manasa , Dhanapal Priyadarshini , Sagar Chandrakant Dalsaniya , Gurrala Sheelu , Thenkrishnan Kumaraguru","doi":"10.1016/j.steroids.2025.109600","DOIUrl":"10.1016/j.steroids.2025.109600","url":null,"abstract":"<div><div>The conversion of 17-oxosteroids to 17β-hydroxysteroids stands as a pivotal process in the synthesis of numerous steroidal drugs and intermediates. This study explored the potential of the strain <em>Priestia aryabhattai</em> (IICT-BC-1279) to catalyze the reduction of the C-17 carbonyl group in androst-4-ene-3,17-dione (AD), resulting in the exclusive production of testosterone (TS) through its 17β-hydroxysteroid dehydrogenase (17β-HSD) activity. Optimal conditions for this reduction were achieved at pH 7.0 and 25 °C, with supplementation of AD as an inducer (0.01 g/L), 1 % Tween 80 (w/v) and ethanol as co-solvent. Under these optimized parameters, 0.5 g/L AD was efficiently converted to TS as a sole product, achieving a yield of > 95 % and diastereomeric excess (<em>d.e</em>) of > 99 % within 48 h. The absence of by-products in this microbial 17β-reduction process simplifies product purification, highlighting the strain’s potential as a valuable biocatalyst for this essential transformation. Additionally, the conversion of androsta-1,4-diene-3,17-dione (ADD) to (+)-Boldenone (BD) was studied to explore substrate scope.</div></div>","PeriodicalId":21997,"journal":{"name":"Steroids","volume":"217 ","pages":"Article 109600"},"PeriodicalIF":2.1,"publicationDate":"2025-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143683854","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-16DOI: 10.1016/j.steroids.2025.109599
Olga Timoshenko , Elena Kugaevskaya , Tatiana Gureeva , Galina Morozevich , Alexey Lupatov , Arif Mekhtiev , Anton Rudovich , Vladimir Zhabinskii , Vladimir Khripach , Andrey Lisitsa
Cervical cancer is the fourth leading cause of cancer death among women worldwide. Matrix metalloproteinases MMP-2 and MMP-9 play a leading role in the processes of invasion and metastasis in cervical cancer. Research on the development of MMP inhibitors not yielded the expected results due to their serious side effects. Study of signaling pathways involved in regulation of MMPs expression is of great importance for search of new classes of therapeutic drugs. Aberrant activation of the Sonic Hedgehog (Shh) signaling pathway is associated with increased MMPs in many types of human cancer. This study investigated the inhibitory action of 17β-((3-butylisoxazol-5-yl)methyl)-androst-5-en-3β-ol on the Shh signaling pathway key genes (Ptch, Smo, Gli) expression and MMP-2, MMP-9 genes expression in human cervical carcinoma cell lines (SiHa and CaSki) and keratinocytes (HaCaT). Cyclopamine was used for comparative analysis. Gene expression analysis was performed using real-time PCR; the effects on survival and cell cycle were studied using the MTT test and flow cytometry method. 17β-((3-butylisoxazol-5-yl)methyl)-androst-5-en-3β-ol had higher cytotoxicity and more effectively blocked the Shh signaling pathway genes and MMP-2 and MMP-9 genes compared to cyclopamine in all cell lines. The results obtained demonstrate potential of 17β-((3-butylisoxazol-5-yl)methyl)-androst-5-en-3β-ol as the anticancer drug that simultaneously block the Shh signaling pathway and MMP expression. We are confident that the search for substances capable of simultaneously affecting several key components involved in tumor progression is of great importance for the creation of next-generation therapeutic agents.
{"title":"Isoxazolyl steroid blocks the Shh signaling pathway and the expression of MMP-2 and MMP-9 in cervical carcinoma cell lines","authors":"Olga Timoshenko , Elena Kugaevskaya , Tatiana Gureeva , Galina Morozevich , Alexey Lupatov , Arif Mekhtiev , Anton Rudovich , Vladimir Zhabinskii , Vladimir Khripach , Andrey Lisitsa","doi":"10.1016/j.steroids.2025.109599","DOIUrl":"10.1016/j.steroids.2025.109599","url":null,"abstract":"<div><div>Cervical cancer is the fourth leading cause of cancer death among women worldwide. Matrix metalloproteinases MMP-2 and MMP-9 play a leading role in the processes of invasion and metastasis in cervical cancer. Research on the development of MMP inhibitors not yielded the expected results due to their serious side effects. Study of signaling pathways involved in regulation of MMPs expression is of great importance for search of new classes of therapeutic drugs. Aberrant activation of the Sonic Hedgehog (Shh) signaling pathway is associated with increased MMPs in many types of human cancer. This study investigated the inhibitory action of 17β-((3-butylisoxazol-5-yl)methyl)-androst-5-en-3β-ol on the Shh signaling pathway key genes (Ptch, Smo, Gli) expression and MMP-2, MMP-9 genes expression in human cervical carcinoma cell lines (SiHa and CaSki) and keratinocytes (HaCaT). Cyclopamine was used for comparative analysis. Gene expression analysis was performed using real-time PCR; the effects on survival and cell cycle were studied using the MTT test and flow cytometry method. 17β-((3-butylisoxazol-5-yl)methyl)-androst-5-en-3β-ol had higher cytotoxicity and more effectively blocked the Shh signaling pathway genes and MMP-2 and MMP-9 genes compared to cyclopamine in all cell lines. The results obtained demonstrate potential of 17β-((3-butylisoxazol-5-yl)methyl)-androst-5-en-3β-ol as the anticancer drug that simultaneously block the Shh signaling pathway and MMP expression. We are confident that the search for substances capable of simultaneously affecting several key components involved in tumor progression is of great importance for the creation of next-generation therapeutic agents.</div></div>","PeriodicalId":21997,"journal":{"name":"Steroids","volume":"217 ","pages":"Article 109599"},"PeriodicalIF":2.1,"publicationDate":"2025-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143658723","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-15DOI: 10.1016/j.steroids.2025.109598
Giuseppe Faita , Mariella Mella , Paolo Quadrelli
Benzo- and anthracenenitrile oxides undergo 1,3-dipolar cycloaddition reactions with 17 α- acetate norethisterone affording the expected isoxazoline and isoxazoles derivatives in good yields and as single regioisomers. The structures of all the new compounds were elucidated on the basis of the corresponding analytical and spectroscopic data, which were presented and discussed. The stereo- and regiochemical outcome of the cycloadditions were also accounted on the basis of 1,3-dipolar cycloaddition theory and computational investigations. Electronic (Frontier Orbital theory) and steric effects are at work in orienting selectively the cycloaddition to specific regioisomeric steroids.
{"title":"Stereo- and regioselectivity in nitrile oxide cycloaddition reactions to norethisterone acetate as dipolarophile","authors":"Giuseppe Faita , Mariella Mella , Paolo Quadrelli","doi":"10.1016/j.steroids.2025.109598","DOIUrl":"10.1016/j.steroids.2025.109598","url":null,"abstract":"<div><div>Benzo- and anthracenenitrile oxides undergo 1,3-dipolar cycloaddition reactions with 17 α- acetate norethisterone affording the expected isoxazoline and isoxazoles derivatives in good yields and as single regioisomers. The structures of all the new compounds were elucidated on the basis of the corresponding analytical and spectroscopic data, which were presented and discussed. The stereo- and regiochemical outcome of the cycloadditions were also accounted on the basis of 1,3-dipolar cycloaddition theory and computational investigations. Electronic (Frontier Orbital theory) and steric effects are at work in orienting selectively the cycloaddition to specific regioisomeric steroids.</div></div>","PeriodicalId":21997,"journal":{"name":"Steroids","volume":"217 ","pages":"Article 109598"},"PeriodicalIF":2.1,"publicationDate":"2025-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143644388","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-09DOI: 10.1016/j.steroids.2025.109597
Dayamin Martínez_Brito , Patrizia Leogrande , Xavier de la Torre , Francesco Botrè
This work studied the short-term stability of 6α-chloro-testosterone (6-CT), 6β-bromo-androstenedione (6-BrAED) and 6-oxo-androstenedione (6-oxo-AED) in methanol (MeOH) and dimethylsulfoxide (DMSO) solutions by gas and liquid chromatography coupled to mass spectrometry.
Solutions of 6-CT, 6-BrAED and 6-oxo-AED were prepared in MeOH and DMSO. They were stored at room temperature, +4°C and −20 °C. Measurements were made at 0, 7, 30, 60 and 90 days after solutions preparation, by liquid chromatography-tandem mass spectrometry and gas chromatography-high resolution mass spectrometry.
Although the degradation of 6-CT and 6-BrAED was extensive, the most notable result was that the higher degradation occurred in DMSO instead of MeOH. The interaction of DMSO with halogenated species and secondary hydroxyl groups favored the degradation of these compounds by forming chemically related species. No degradation of 6-oxo-AED in either MeOH or DMSO was observed.
Pronounced degradation of the 6-CT and 6-BrAED, during the derivatization reaction for the gas chromatography-mass spectrometry analysis was observed. Because of the acidic condition of the reaction and depending on the reactant, it was favored the loss of the halogen molecule or the dehydration reaction to form the unsaturated (Δ6) steroid derivative.
Our finding suggests to take duly into account the possibility of degradation processes when performing quantitative determination of 6-CT, 6-BrAED and 6-oxo-AED by chromatographic-spectrometric techniques based on the use of reference solutions stored for sufficiently long times.
{"title":"Evaluation of the short-term stability of 6α-chloro-testosterone, 6β-bromo-androstenedione and 6-oxo-androstenedione in dimethylsulfoxide and methanol using liquid and gas chromatography − mass spectrometry","authors":"Dayamin Martínez_Brito , Patrizia Leogrande , Xavier de la Torre , Francesco Botrè","doi":"10.1016/j.steroids.2025.109597","DOIUrl":"10.1016/j.steroids.2025.109597","url":null,"abstract":"<div><div>This work studied the short-term stability of 6α-chloro-testosterone (6-CT), 6β-bromo-androstenedione (6-BrAED) and 6-oxo-androstenedione (6-oxo-AED) in methanol (MeOH) and dimethylsulfoxide (DMSO) solutions by gas and liquid chromatography coupled to mass spectrometry.</div><div>Solutions of 6-CT, 6-BrAED and 6-oxo-AED were prepared in MeOH and DMSO. They were stored at room temperature, +4°C and −20 °C. Measurements were made at 0, 7, 30, 60 and 90 days after solutions preparation, by liquid chromatography-tandem mass spectrometry and gas chromatography-high resolution mass spectrometry.</div><div>Although the degradation of 6-CT and 6-BrAED was extensive, the most notable result was that the higher degradation occurred in DMSO instead of MeOH. The interaction of DMSO with halogenated species and secondary hydroxyl groups favored the degradation of these compounds by forming chemically related species. No degradation of 6-oxo-AED in either MeOH or DMSO was observed.</div><div>Pronounced degradation of the 6-CT and 6-BrAED, during the derivatization reaction for the gas chromatography-mass spectrometry analysis was observed. Because of the acidic condition of the reaction and depending on the reactant, it was favored the loss of the halogen molecule or the dehydration reaction to form the unsaturated (Δ6) steroid derivative.</div><div>Our finding suggests to take duly into account the possibility of degradation processes when performing quantitative determination of 6-CT, 6-BrAED and 6-oxo-AED by chromatographic-spectrometric techniques based on the use of reference solutions stored for sufficiently long times.</div></div>","PeriodicalId":21997,"journal":{"name":"Steroids","volume":"217 ","pages":"Article 109597"},"PeriodicalIF":2.1,"publicationDate":"2025-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143606530","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-03DOI: 10.1016/j.steroids.2025.109589
Qing-Jiang Xu , Jia-Chen Liu , Jia Xu , Xin Wang , Xiao-Ya Shang , Jiachen Zi
Three new diterpenoids, 12,16-epoxy-11-hydroxy-17(15 → 16)-abeo-abieta-8,11,13-trien-7-one (1), 7,12-dihydroxy abieta-6,8,10(5),11,13-quien-20-oic acid 1,20-lactone (2), labda-5(10),13(E)-dien-15-ol (11), one new natural product (2E,6E)-3,7-dimethyl-9-[(1S,6R)-1,2,6-trimethylcyclohex-2-enyl]nona-2,6-dien-1-ol (16) and thirteen known compounds were isolated and elucidated from the excellent antimicrobial active fractions of Rosmarinus officinalis ethanol extract. The structures of the isolated compounds were determined by spectral data analysis and combined with literature reports. Among them, monocyclic diterpenoids (16 and 17) were discovered from rosemary for the first time. All isolated compounds were tested for antimicrobial activity against four strains (B. subtilis, S. aureus, P. aeruginosa, and Fusarium spp.), with six compounds showing very strong inhibitory activity against B. subtilis and four compounds showing strong inhibitory activity against P. aeruginosa.
{"title":"Antimicrobial diterpenoids from Rosmarinus officinalis","authors":"Qing-Jiang Xu , Jia-Chen Liu , Jia Xu , Xin Wang , Xiao-Ya Shang , Jiachen Zi","doi":"10.1016/j.steroids.2025.109589","DOIUrl":"10.1016/j.steroids.2025.109589","url":null,"abstract":"<div><div>Three new diterpenoids, 12,16-epoxy-11-hydroxy-17(15 → 16)-<em>abeo</em>-abieta-8,11,13-trien-7-one (<strong>1</strong>), 7,12-dihydroxy abieta-6,8,10(5),11,13-quien-20-oic acid 1,20-lactone (<strong>2</strong>), labda-5(10),13(<em>E)</em>-dien-15-ol (<strong>11</strong>), one new natural product (2<em>E</em>,6<em>E</em>)-3,7-dimethyl-9-[(1<em>S</em>,6<em>R</em>)-1,2,6-trimethylcyclohex-2-enyl]nona-2,6-dien-1-ol (<strong>16</strong>) and thirteen known compounds were isolated and elucidated from the excellent antimicrobial active fractions of <em>Rosmarinus officinalis</em> ethanol extract. The structures of the isolated compounds were determined by spectral data analysis and combined with literature reports. Among them, monocyclic diterpenoids (<strong>16</strong> and <strong>17</strong>) were discovered from rosemary for the first time. All isolated compounds were tested for antimicrobial activity against four strains (<em>B. subtilis, S. aureus</em>, <em>P. aeruginosa</em>, and <em>Fusarium spp</em>.), with six compounds showing very strong inhibitory activity against <em>B. subtilis</em> and four compounds showing strong inhibitory activity against <em>P. aeruginosa</em>.</div></div>","PeriodicalId":21997,"journal":{"name":"Steroids","volume":"217 ","pages":"Article 109589"},"PeriodicalIF":2.1,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143563327","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-01DOI: 10.1016/j.steroids.2025.109587
Lili Ye , Ruiyan Wang , Jun Zhao , Jingrong Chen , Feng Wang
Introduction
Cardiac endogenous senescence will gradually change and aggravate with age. Recent research showed that 17β-estradiol (17β-E2), an estrogen with numerous biological activities including the prevention of vascular senescence. However, how 17β-E2 against cardiac aging is still unknown. This work addressed the underlying mechanism with regard to Beclin1 and autophagy activity to better understand the anti-senescent effect of 17β-E2 on a well-established animal model of cardiac aging.
Material and methods
In this study, an aging model in female mice was established using d-galactose and ovariectomy. Cardiac function was evaluated by echocardiography, RNA-seq was performed to analyze the gene expression profiles of myocardial tissues from 17β-E2 treated mice. Additionally,The levels of Beclin1, LC3, P62, and ATG5 in myocardial tissues were assessed using qPCR and Western blotting. Methylation levels of the Beclin1 promoter region in myocardial tissues were determined by MSP and BSP.
Results
The findings demonstrated that cardiac aging mice treated with 17β-E2 had improved heart function. 17β-E2 restored EF(increase 1.25-fold) and FS(increase 1.2-fold) to near-normal levels. By RNA-sequencing and Gene Set Enrichment Analysis (GSEA) analysis, the autophagy signaling pathway was further enriched in the myocardial tissue of cardiac aging mice treated with 17β-E2, and we also discovered that 17β-E2 suppress the methylation of Beclin1 promoter region, which mediate the activation of autophagy signal.
Conclusions
Overall, our data showed that 17β-E2′s anti-senescent effect on cardiac aging mice was mediated by the crucial suppression of methylation in the Beclin1 promoter area and subsequent activation of the autophagy signal, which may present a possible therapeutic approach to prevent cardiac aging.
心脏内源性衰老会随着年龄的增长而逐渐改变和加剧。最近的研究表明，17β-雌二醇（17β-E2）是一种具有多种生物活性的雌激素，包括预防血管衰老。然而，17β-E2如何对抗心脏老化仍是未知的。为了更好地了解17β-E2在心脏衰老动物模型中的抗衰老作用，本研究探讨了Beclin1和自噬活性的潜在机制。材料与方法：本研究采用d-半乳糖和卵巢切除术建立雌性小鼠衰老模型。用超声心动图评价心脏功能，用RNA-seq分析17β-E2处理小鼠心肌组织的基因表达谱。采用qPCR和Western blotting检测心肌组织Beclin1、LC3、P62、ATG5的表达水平。心肌组织中Beclin1启动子区甲基化水平通过MSP和BSP检测。结果：心脏老化小鼠经17β-E2处理后，心脏功能得到改善。17β-E2恢复EF（增加1.25倍）和FS（增加1.2倍）至接近正常水平。通过rna测序和基因集富集分析（Gene Set Enrichment Analysis， GSEA）分析，我们发现17β-E2在心脏衰老小鼠心肌组织中进一步富集了自噬信号通路，并且我们还发现17β-E2抑制介导自噬信号激活的Beclin1启动子区甲基化。结论：总体而言，我们的数据表明，17β-E2对心脏衰老小鼠的抗衰老作用是通过关键的Beclin1启动子区甲基化抑制和随后的自噬信号激活介导的，这可能是一种可能的预防心脏衰老的治疗方法。
{"title":"17β-estradiol delays cardiac aging through suppressing the methylation of Beclin1 in a murine model","authors":"Lili Ye , Ruiyan Wang , Jun Zhao , Jingrong Chen , Feng Wang","doi":"10.1016/j.steroids.2025.109587","DOIUrl":"10.1016/j.steroids.2025.109587","url":null,"abstract":"<div><h3>Introduction</h3><div>Cardiac endogenous senescence will gradually change and aggravate with age. Recent research showed that 17β-estradiol (17β-E2), an estrogen with numerous biological activities including the prevention of vascular senescence. However, how 17β-E2 against cardiac aging is still unknown. This work addressed the underlying mechanism with regard to Beclin1 and autophagy activity to better understand the anti-senescent effect of 17β-E2 on a well-established animal model of cardiac aging.</div></div><div><h3>Material and methods</h3><div>In this study, an aging model in female mice was established using <span>d</span>-galactose and ovariectomy. Cardiac function was evaluated by echocardiography, RNA-seq was performed to analyze the gene expression profiles of myocardial tissues from 17β-E2 treated mice. Additionally,The levels of Beclin1, LC3, <em>P</em>62, and ATG5 in myocardial tissues were assessed using qPCR and Western blotting. Methylation levels of the Beclin1 promoter region in myocardial tissues were determined by MSP and BSP.</div></div><div><h3>Results</h3><div>The findings demonstrated that cardiac aging mice treated with 17β-E2 had improved heart function. 17β-E2 restored EF(increase 1.25-fold) and FS(increase 1.2-fold) to near-normal levels. By RNA-sequencing and Gene Set Enrichment Analysis (GSEA) analysis, the autophagy signaling pathway was further enriched in the myocardial tissue of cardiac aging mice treated with 17β-E2, and we also discovered that 17β-E2 suppress the methylation of Beclin1 promoter region, which mediate the activation of autophagy signal.</div></div><div><h3>Conclusions</h3><div>Overall, our data showed that 17β-E2′s anti-senescent effect on cardiac aging mice was mediated by the crucial suppression of methylation in the Beclin1 promoter area and subsequent activation of the autophagy signal, which may present a possible therapeutic approach to prevent cardiac aging.</div></div>","PeriodicalId":21997,"journal":{"name":"Steroids","volume":"216 ","pages":"Article 109587"},"PeriodicalIF":2.1,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143543073","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-01DOI: 10.1016/j.steroids.2025.109575
Kamlesh Sharma, Priyanka
The reaction mechanism of the formation of pyridyl-cholestane derivative 4 from a multi-component reaction involving cholestane-6-one, aromatic aldehyde, malononitrile, and ammonium acetate in presence of magnesium oxide nanoparticles (MgO NPs) as catalyst, was studied successfully by using DFT calculations. The mechanism involved condensation, cyclization, and aromatization steps which were investigated successfully theoretically. The theoretical calculations of physicochemical parameters, including Gibbs free energy, frontier molecular orbitals (FMOs), dipole moments, and hardness, of all the intermediates and transition states molecules. The study revealed the formation of key intermediates and transition states, with detailed analysis of their stability and electronic structures.
The reaction pathway begins with the formation of enamine I and α,β-unsaturated nitrile II, followed by Michael addition to produce intermediate B. The cyclization of A to intermediate B, which has the highest activation energy barrier was identified as slowest and the rate-determining step. The following steps, including cyclization (B to C) and proton transfer (C to D), exhibit progressively lower activation barriers and enhanced stability. Theoretical analysis indicates that the reaction is thermodynamically favorable, as the product is more stable than the initial reactants.
This study highlights the mechanistic insights contributing to the understanding of multi-component reactions in organic synthesis involved effectiveness of MgO NPs as a heterogeneous catalyst in enabling the efficient synthesis of pyridyl-cholestane derivative 4.
{"title":"Theoretical study of the pyridyl-cholestane formation pathway using DFT: A stepwise mechanistic approach","authors":"Kamlesh Sharma, Priyanka","doi":"10.1016/j.steroids.2025.109575","DOIUrl":"10.1016/j.steroids.2025.109575","url":null,"abstract":"<div><div>The reaction mechanism of the formation of pyridyl-cholestane derivative <strong>4</strong> from a multi-component reaction involving cholestane-6-one, aromatic aldehyde, malononitrile, and ammonium acetate in presence of magnesium oxide nanoparticles (MgO NPs) as catalyst, was studied successfully by using DFT calculations. The mechanism involved condensation, cyclization, and aromatization steps which were investigated successfully theoretically. The theoretical calculations of physicochemical parameters, including Gibbs free energy, frontier molecular orbitals (FMOs), dipole moments, and hardness, of all the intermediates and transition states molecules. The study revealed the formation of key intermediates and transition states, with detailed analysis of their stability and electronic structures.</div><div>The reaction pathway begins with the formation of enamine <strong>I</strong> and α,β-unsaturated nitrile <strong>II</strong>, followed by Michael addition to produce intermediate <strong>B</strong>. The cyclization of <strong>A</strong> to intermediate <strong>B</strong>, which has the highest activation energy barrier was identified as slowest and the rate-determining step. The following steps, including cyclization (<strong>B</strong> to <strong>C</strong>) and proton transfer (<strong>C</strong> to <strong>D</strong>), exhibit progressively lower activation barriers and enhanced stability. Theoretical analysis indicates that the reaction is thermodynamically favorable, as the product is more stable than the initial reactants.</div><div>This study highlights the mechanistic insights contributing to the understanding of multi-component reactions in organic synthesis involved effectiveness of MgO NPs as a heterogeneous catalyst in enabling the efficient synthesis of pyridyl-cholestane derivative 4.</div></div>","PeriodicalId":21997,"journal":{"name":"Steroids","volume":"216 ","pages":"Article 109575"},"PeriodicalIF":2.1,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143543248","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-28DOI: 10.1016/j.steroids.2025.109584
Mufeda Ahmed Hazea Gazaem , Wan Nurul Nazneem Wan Othman , Syed Adnan Ali Shah , Mustapha Salihu , Azeana Zahari , Siti Hajar Sadiran , Fatimah Salim
Biotransformations catalysed by microbes are promising approach for producing a vast library of structurally diverse chemical molecules with applications in the pharmaceutical industry. The biotransformation of 11α-acetoxyprogesterone (1) by Phyllosticta sp. 16L1 has not been previously reported. In this study, the biotransformation of 11α-acetoxyprogesterone (1) was performed for the first time using the Phyllosticta sp. 16L1 strain. After an 8-day fermentation period, a new biotransformation metabolite, named as 11α-acetoxy-16α-hydroxyprogesterone (16α-hydroxy-3,20-dioxopregn-4-en-11α-yl acetate) (2) was isolated from the culture broth, along with its known isomer, 11α-acetoxy-15α-hydroxyprogesterone (3). The structure determination of the biotransformed products relied on comprehensive spectroscopic data, encompassing 1D and 2D-NMR, as well as LCMS analyses. The cytotoxic activity of the two biotransformed metabolites was assessed against selective human cancer cell lines, including hepatocellular carcinoma (HepG2), triple-negative breast cancer (MDA-MB-231), colorectal adenocarcinoma (Caco-2), and lung adenocarcinoma (A549). The results demonstrated that both metabolites 2 and 3 exhibited cytotoxic effects on the evaluated cell lines. Metabolite 2 showed stronger cytotoxic potential, with IC50 values ranging from 6.65 to 27.75 μM, while metabolite 3 displayed lower potency, with IC50 values between 38.20 and 162.53 μM. Notably, both metabolites exhibited minimal toxicity towards the normal liver Chang cells. Molecular docking studies were conducted to predict the binding modes and affinities of the metabolites against two targets (PDB: 5EM8 and 6V6O), both in 2D and 3D representations, with binding energies ranging from −8.5 to −7.2 kcal/mol. The results revealed that metabolites 2 and 3 interacted with key clinically significant amino acid residues, Lys745 and Met793, through conventional hydrogen bonding.
{"title":"New hydroxylated metabolite derived from the microbial biotransformation of 11α-acetoxyprogesterone by the endophytic fungus Phyllosticta sp. 16L1 and its cytotoxic activity","authors":"Mufeda Ahmed Hazea Gazaem , Wan Nurul Nazneem Wan Othman , Syed Adnan Ali Shah , Mustapha Salihu , Azeana Zahari , Siti Hajar Sadiran , Fatimah Salim","doi":"10.1016/j.steroids.2025.109584","DOIUrl":"10.1016/j.steroids.2025.109584","url":null,"abstract":"<div><div>Biotransformations catalysed by microbes are promising approach for producing a vast library of structurally diverse chemical molecules with applications in the pharmaceutical industry. The biotransformation of 11α-acetoxyprogesterone (<strong>1</strong>) by <em>Phyllosticta</em> sp. 16L1 has not been previously reported. In this study, the biotransformation of 11α-acetoxyprogesterone (<strong>1</strong>) was performed for the first time using the <em>Phyllosticta</em> sp. 16L1 strain. After an 8-day fermentation period, a new biotransformation metabolite, named as 11α-acetoxy-16α-hydroxyprogesterone (16α-hydroxy-3,20-dioxopregn-4-en-11α-yl acetate) (<strong>2</strong>) was isolated from the culture broth, along with its known isomer, 11α-acetoxy-15α-hydroxyprogesterone (<strong>3</strong>). The structure determination of the biotransformed products relied on comprehensive spectroscopic data, encompassing 1D and 2D-NMR, as well as LCMS analyses. The cytotoxic activity of the two biotransformed metabolites was assessed against selective human cancer cell lines, including hepatocellular carcinoma (HepG2), triple-negative breast cancer (MDA-MB-231), colorectal adenocarcinoma (Caco-2), and lung adenocarcinoma (A549). The results demonstrated that both metabolites <strong>2</strong> and <strong>3</strong> exhibited cytotoxic effects on the evaluated cell lines. Metabolite <strong>2</strong> showed stronger cytotoxic potential, with IC<sub>50</sub> values ranging from 6.65 to 27.75 μM, while metabolite <strong>3</strong> displayed lower potency, with IC<sub>50</sub> values between 38.20 and 162.53 μM. Notably, both metabolites exhibited minimal toxicity towards the normal liver Chang cells. Molecular docking studies were conducted to predict the binding modes and affinities of the metabolites against two targets (PDB: 5EM8 and 6V6O), both in 2D and 3D representations, with binding energies ranging from −8.5 to −7.2 kcal/mol. The results revealed that metabolites <strong>2</strong> and <strong>3</strong> interacted with key clinically significant amino acid residues, Lys745 and Met793, through conventional hydrogen bonding.</div></div>","PeriodicalId":21997,"journal":{"name":"Steroids","volume":"216 ","pages":"Article 109584"},"PeriodicalIF":2.1,"publicationDate":"2025-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143537733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-28DOI: 10.1016/j.steroids.2025.109588
Siqi Xu , Ling Fang , Jianfeng Cai , Shuopo Fang , Huide Zhu , Fei Lin , Xiaorui Cai
A new family of steroidal compounds based on a heteroaryl-4,5-dihydropyrazole thiazolinone core structure was designed and synthesized through structural modifications. The anti-neuroinflammatory activity of these compounds was evaluated in lipopolysaccharide (LPS)-stimulated murine microglial BV-2 cells in vitro. Among the synthesized compounds, 10b and 10d effectively inhibited nitric oxide (NO) production, with compound 10b emerging as the most potent anti-neuroinflammatory agent (IC50 = 2.05 μM). Compound 10b demonstrated significantly greater inhibitory effects than progesterone (prog) (IC50 = 3.23 μM) and reduced NO production in a concentration-dependent manner. Furthermore, compound 10b attenuated the release of pro-inflammatory mediators, including tumour necrosis factor (TNF)-α, interleukin-1β (IL-1β), interleukin-6 (IL-6), and prostaglandin E2 (PGE2). It also inhibited the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Mechanistic studies revealed that compound 10b significantly suppressed the transcriptional activity of nuclear factor kappa B (NF-κB) in activated microglial cells and prevented NF-κB p65 activation and IκBα degradation. These effects were likely mediated by the inhibition of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) signaling pathways. Additionally, molecular docking studies suggested that the anti-neuroinflammatory effects of compound 10b may result from its hydrophobic and hydrophilic interactions with iNOS and COX-2, supporting its proposed mechanism of action. In summary, these findings suggest that compound 10b exerts anti-neuroinflammatory effects in LPS-stimulated BV-2 microglial cells by modulating key inflammatory pathways, including NF-κB and MAPK signaling.
{"title":"Design and discovery of novel heteroaryl substituted pregnenolone derivatives as potent anti-neuroinflammatory agents targeting LPS-stimulated BV-2 microglial cells","authors":"Siqi Xu , Ling Fang , Jianfeng Cai , Shuopo Fang , Huide Zhu , Fei Lin , Xiaorui Cai","doi":"10.1016/j.steroids.2025.109588","DOIUrl":"10.1016/j.steroids.2025.109588","url":null,"abstract":"<div><div>A new family of steroidal compounds based on a heteroaryl-4,5-dihydropyrazole thiazolinone core structure was designed and synthesized through structural modifications. The anti-neuroinflammatory activity of these compounds was evaluated in lipopolysaccharide (LPS)-stimulated murine microglial BV-2 cells <em>in vitro</em>. Among the synthesized compounds, <strong>10b</strong> and <strong>10d</strong> effectively inhibited nitric oxide (NO) production, with compound <strong>10b</strong> emerging as the most potent anti-neuroinflammatory agent (IC<sub>50</sub> = 2.05 μM). Compound <strong>10b</strong> demonstrated significantly greater inhibitory effects than progesterone (<strong>prog</strong>) (IC<sub>50</sub> = 3.23 μM) and reduced NO production in a concentration-dependent manner. Furthermore, compound <strong>10b</strong> attenuated the release of pro-inflammatory mediators, including tumour necrosis factor (TNF)-α, interleukin-1β (IL-1β), interleukin-6 (IL-6), and prostaglandin E2 (PGE2). It also inhibited the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Mechanistic studies revealed that compound <strong>10b</strong> significantly suppressed the transcriptional activity of nuclear factor kappa B (NF-κB) in activated microglial cells and prevented NF-κB p65 activation and IκBα degradation. These effects were likely mediated by the inhibition of c-Jun <em>N</em>-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) signaling pathways. Additionally, molecular docking studies suggested that the anti-neuroinflammatory effects of compound <strong>10b</strong> may result from its hydrophobic and hydrophilic interactions with iNOS and COX-2, supporting its proposed mechanism of action. In summary, these findings suggest that compound <strong>10b</strong> exerts anti-neuroinflammatory effects in LPS-stimulated BV-2 microglial cells by modulating key inflammatory pathways, including NF-κB and MAPK signaling.</div></div>","PeriodicalId":21997,"journal":{"name":"Steroids","volume":"216 ","pages":"Article 109588"},"PeriodicalIF":2.1,"publicationDate":"2025-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143534689","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-26DOI: 10.1016/j.steroids.2025.109583
Junwei Wang , Shuhui Wang , Meng Wang , Jinfei Yang
Non-obstructive azoospermia (NOA) is the most common cause of male infertility, accounting for approximately 60 % of azoospermia cases. In recent years, gene mutations have emerged as the primary factor under investigation for the etiology of NOA. Therefore, finding the cause and pathogenesis of NOA at the genetic level has become one of the current research hotspots. Genetic analysis of NOA patients revealed that gene mutations primarily concentrate in protein-coding regions and non-coding RNAs, predominantly occurring in cases of non-obstructive azoospermia. Hence, understanding the relationship between these gene mutations and NOA can not only provide new ideas for treatment, but also provide a theoretical basis for revealing the pathogenesis of NOA. This article comprehensively reviews recent advancements in identifying genes that are intricately associated with azoospermia. These results will provide meaningful guidance for the future development of NOA-targeted therapeutic drugs.
{"title":"Analysis of genes implicated in non-obstructive azoospermia","authors":"Junwei Wang , Shuhui Wang , Meng Wang , Jinfei Yang","doi":"10.1016/j.steroids.2025.109583","DOIUrl":"10.1016/j.steroids.2025.109583","url":null,"abstract":"<div><div>Non-obstructive azoospermia (NOA) is the most common cause of male infertility, accounting for approximately 60 % of azoospermia cases. In recent years, gene mutations have emerged as the primary factor under investigation for the etiology of NOA. Therefore, finding the cause and pathogenesis of NOA at the genetic level has become one of the current research hotspots. Genetic analysis of NOA patients revealed that gene mutations primarily concentrate in protein-coding regions and non-coding RNAs, predominantly occurring in cases of non-obstructive azoospermia. Hence, understanding the relationship between these gene mutations and NOA can not only provide new ideas for treatment, but also provide a theoretical basis for revealing the pathogenesis of NOA. This article comprehensively reviews recent advancements in identifying genes that are intricately associated with azoospermia. These results will provide meaningful guidance for the future development of NOA-targeted therapeutic drugs.</div></div>","PeriodicalId":21997,"journal":{"name":"Steroids","volume":"216 ","pages":"Article 109583"},"PeriodicalIF":2.1,"publicationDate":"2025-02-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143519861","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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