Parkinson's disease (PD) ranks as the second most prevalent neurodegenerative disorder but is still devoid of neuroprotective treatment. Although approaches with disease modifying ability along with symptomatic relief has become an utmost necessity, the multifactorial nature of PD presents challenges for efficacy evaluation of potential test compound. This study attempts to address these issues by employing a rotenone induced PD model involving intranigral rotenone injection for evaluation of the neuroprotective efficacy of Daidzein (DZ) a soy isoflavone and a phytoestrogen. In this study, male Sprague Dawley rats after bilateral intranigral rotenone (12 μg) injection, were treated with DZ at a dose of 5, 10 and 20 mg/kg for 30 days. The neurobehavioral evaluation comprised of Rota-rod, Open field and Barnes maze test. The biochemical analysis constituting oxidative stress (Reduced glutathione, superoxide dismutase, catalase and lipid peroxidation), inflammation (TNF-α), mitochondrial alteration (complex I activity and biogenesis) was conducted on mid-brain tissue after 30 days of treatment. The Substantia nigra and striatum were subjected to immunohistochemical analysis (IHC) for TH positive neurons and Glial Fibrillary Acidic Protein. The analysis revealed significant improvement by daidzein in motor co-ordination and attenuation in cognitive deficits due to rotenone. The biochemical assessment exhibited significant decrement in oxidative stress as well as inflammation. DZ treatment also prevented complex I inhibition and promoted mitochondrial biogenesis eventually contributing to the neuroprotection apparent in IHC. Thus, the results strongly corroborate the neuroprotective potential of DZ against rotenone induced model of PD.

Bacterial biofilms are complex, organized structures that adhere to surfaces, protected by a self-produced extracellular matrix. This conformation makes it difficult to eradicate infections with conventional treatments. In view of this, natural compounds have emerged as promising therapeutic alternatives. This study evaluated the potential of betulinic acid, a pentacyclic triterpene, in inhibiting the formation and eradication of bacterial biofilms, as well as predicting its pharmacokinetic properties and toxicity by means of in silico analyses. Six bacterial strains were tested. The crystal violet test was used to evaluate the antibiofilm activity, with chlorhexidine and the antibiotics norfloxacin, ampicillin and gentamicin as controls. The results showed that betulinic acid had moderate to good activity in inhibiting biofilm formation and a variable response in eradication, depending on the strain. In silico analyses indicated favorable physicochemical and pharmacokinetic properties, with emphasis on good intestinal absorption, oral bioavailability and absence of inhibition of cytochrome P450 enzymes, although potential toxicity risks were identified. These findings suggest that betulinic acid is a promising candidate for the development of new therapeutic strategies to combat infections associated with biofilms.

A concise synthesis of brassinolide and castasterone has been accomplished herein. The strategy relies on the addition of α-silyl substituted vinyl carbanions to steroidal C-22 aldehydes with pre-formed functionalities in the cyclic part. Commercially available brassinosteroids were used as a starting material. The key step of the present synthesis was the opening of the 23,24-epoxide with MeAl in the presence of PhP as a Lewis base, which enhances the formation of a more nucleophilic alkylating agent. It was shown why the use of MeAl is incompatible with the 3α,5-cyclo steroids, which are often employed as intermediates in the synthesis of brassinosteroids.

Glucocorticoid-induced osteoporosis (GIOP) is a leading cause of secondary osteoporosis (OP). β-Ecdysone (βEcd), a naturally occurring estrogen analog, was evaluated for its ability to mitigate the effects of glucocorticoids (GCs) on osteocytes, the crucial cells in bone remodeling. In GIOP mouse model induced by dexamethasone (Dex), micro-CT, biomechanical testing, silver nitrate staining, and hematoxylin-eosin (HE) staining were employed, demonstrating that βEcd effectively attenuated Dex-induced decreases in bone mass and strength, and alleviated Dex induced reduction in osteocyte dendrite and viability. Network pharmacology analysis predicted that the therapeutic efficacy of βEcd against GIOP is mediated through the crucial targets such as protein kinase B (Akt1), with significant enrichment in pathways including apoptosis and phosphoinositide 3-kinase (PI3K)-Akt signaling. In vitro, the osteocyte-like MLO-Y4 cells were treated with 10 μM Dex for 48 h in the presence or absence of βEcd or the PI3K inhibitor LY294002 (LY). Crystal violet staining and connexin43 (CX43) immunofluorescence (IF) staining were employed, and western blot was used to assess the levels of Akt1, phospho (p)-Akt, CX43, p-CX43, and apoptosis-related factors. Hoechst staining and annexin V/PI apoptosis assays were also used to evaluate apoptosis. The results demonstrated that βEcd counteracted Dex-induced apoptosis by modulating Akt1 and CX43 expression in MLO-Y4 cells, while inhibition of Akt activity reversed these effects, suggesting that βEcd targets the Akt1 gene. The findings indicate that βEcd protects osteocytes from GC-induced apoptosis through Akt-mediated regulation of CX43, highlighting its potential as a therapeutic approach for the prevention and treatment of GIOP.

Solanum viarum is a valuable medicinal plant native to India and widely distributed throughout Asia. It serves as a commercially viable raw source for the steroidal drug industry, being the richest natural source of an important steroidal alkaloid- solasodine. Enhancement of solasodine content in in vitro plant cultures is always a keen interest for tissue culturists for the research and development in pharmaceutical industries. In the present study, a multiple linear regression (MLR) model was employed to investigate the synergistic effects of key nutrient components in the growth medium (Mg, Ca, Fe, N, and sucrose) for optimized growth and solasodine production in in vitro plant cultures of S. viarum. All the cultures were harvested after 35, 45, and 55 days of the culture cycle. The three models were designed to predict growth index, solasodine content, and solasodine yield in the plant cultures. The designed model was further evaluated by performing a validation experiment using ten different experimental setups, which showed a greater similarity between the predicted and experimental datasets. The results of the model-based experimental set showed that 1.4 mM Mg, 2.9 mM Ca, 1.9 µM Fe, 41.9 mM Nitrogen, and 4 % (w/v) sucrose resulted in achieving maximum solasodine yield (108.82 mg/g DW) in the plant culture of S. viarum after 54 days of harvest. The proposed MLR model offers a robust and reliable approach for predicting the optimal concentrations of micro- and macronutrients to maximize growth and solasodine accumulation in in vitro cultures of Solanum viarum. This study establishes a strategic framework that can be leveraged to enhance biomass production and secondary metabolite yield, contributing significantly to the large-scale cultivation and pharmaceutical exploitation of this valuable medicinal plant.

Progesterone (P4) is an essential steroid hormone synthesised by the placenta required for the maintenance of pregnancy. In humans, the metabolism of P4 has been implicated in functional P4 withdrawal prior to parturition. We have developed a novel liquid chromatography tandem mass spectrometry (LC-MS/MS) method for quantitation in human serum of pregnenolone, P4 and its four primary reduced metabolites; 20α-hydroxyprogesterone (20α-OHP), 20β-hydroxyprogesterone (20β-OHP), 5α-dihydroprogesterone (5α-DHP) and 5β-dihydroprogesterone (5β-DHP). Following solid phase extraction, chromatographic baseline separation of each steroid was achieved using a biphenyl stationary phase within a 10.0 min runtime, followed by MS detection on a Sciex 6500+. The LC-MS/MS method was validated in accordance with published guidelines, confirming acceptable analytical performance pertaining to linearity, imprecision, accuracy, sensitivity, matrix effects, specificity and carryover. The method was applied to a large cohort of third trimester pregnancies with verified uncomplicated neonatal outcomes. Maternal circulating concentrations of P4, 20α-OHP, 20β-OHP, and 5α-DHP positively correlated with fetal gestational age. The ratio of P4:20β-OHP declined significantly throughout the third trimester, whilst the ratio of P4:5β-DHP increased at full term from 40 weeks' gestation. These findings may indicate a substantive role for β-reduction of P4 in the mechanics of functional P4 withdrawal, either via depletion of the overall pool of bioactive P4 or competitive binding of these metabolites to the P4 receptor in maternal and fetal tissue. Additionally, detailed characterisation of the normal maternal steroidome will facilitate the study of dysregulated placental steroidogenesis, which has been implicated in the pathogenesis of the major obstetric syndromes causing poor pregnancy outcomes.

Advances in analytical techniques have markedly improved the detection of anabolic-androgenic steroids (AAS) in doping controls. This represents a major success for anti-doping efforts, but at the same times raises concerns about inadvertent exposure - not limited to oral ingestion, but also via dermal contact or mucosal absorption. Evaluating the plausibility of such scenarios can be complex, not least because these routes of drug intake can also be deliberately used for doping purposes. Analytical data on transdermal or mucosal absorption and elimination of AAS can be valuable here, particularly regarding the metabolite pattern in urine samples that might help to assess the route of drug delivery. This study investigated the urinary excretion profile of trenbolone and its metabolites after oral, transdermal and buccal administration in healthy male volunteers. Urine samples collected over a period of one week following the administration of 10 mg of trenbolone were analyzed by LC-HRMS, with regard to potential differences in the metabolite profile across the different routes of drug intake. Major trenbolone metabolites were detectable in the urine samples of all participants, however the detection windows varied strongly: Buccal resorption resulted in faster elimination, whereas transdermal application resulted in prolonged detectability, compared to oral administration. A cysteine-conjugated metabolite (M15-cys) was identified that showed higher abundances following transdermal and buccal administration, suggesting its potential as a marker for non-oral intake. In light of the growing importance of transdermal uptake of doping agents, these results provide valuable data for interpreting doping control findings, potentially also relevant for structurally related AAS.

Dehydroepiandrosterone (DHEA) is the most abundant steroid hormone in primates; however, its levels decline with age in both men and women. Numerous studies have documented diverse biological activities of DHEA. This article reviews the current understanding of how DHEA regulates the contractile function of vascular and nonvascular smooth muscle (SM), specifically examining its relaxant properties mediated through nongenomic mechanisms. It summarizes both older and recent findings on DHEA and its role in regulating SM contractile activity. Moreover, it also discusses the potential mechanisms underlying its relaxation effects, including the structural-functional differences in the DHEA molecule and its ability to induce relaxation in various types of SM. Overall, DHEA introduces a novel aspect to the regulation of functional processes involved in SM contractile activity.

Genetics, inflammation, and nutrition contribute to the pathogenesis of metabolic dysfunction-associated steatotic liver disease (MASLD).

Among the phase-I reactions of 6-chloro-testosterone (6-CT), the reductive and oxidative dehalogenation, as well as the dehydrogenation, have been described here. One of the metabolic products could be the 6-ene-testosterone (6-ene-T). The goal of this work was to study the in vitro metabolism of 6-CT and 6-ene-T after incubation with human liver microsomes (HLM), analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and gas chromatography-time-of-flight-mass spectrometry (GC-TOF). 6-CT showed a more extensive metabolism compared to 6-ene-T, with the formation of a larger number of metabolites. In contrast, 6-ene-T, which potentially preserves the C6C7 double bond in most of its metabolites, showed fewer metabolites. Therefore, after incubation with HLM, the main metabolic routes of 6-ene-T are the oxidation of the hydroxyl group on C17β, retaining the double bond C6C7 to form 6-ene-androstenedione, and the hydroxylation on C6. On the other hand 6-CT metabolism produced chlorinated and non-chlorinated metabolites as a result of the phase I reactions which included oxidation, reduction, hydroxylation and (oxidative and reductive) dehalogenation. Specifically for 6-CT, it should be considered that the introduction of a chlorine atom in the 6α position of T may lead to a shift in the metabolic pathway, favoring the formation of 5β-reduced metabolites over 5α-reduced metabolites, due to the steric and electronic effects of the chlorine atom on the interaction between the steroid molecule and the 5α-reductase enzyme.

Betulinic acid (BA) is a pentacyclic triterpene compound with various biological activities. Herein, we designed and synthesized a series of dihydrobetulinic acid (DHBA) and its derivatives for the discovery of potent ecto-5'-nucleotidase (CD73) inhibitors. Biological evaluation of DHBA and its derivatives led to the disclosure of three active compounds DHBA, ZM792 and ZM905. Further investigation of antitumor immunity revealed that DHBA could effectively restore the function of CD4 T cells. These results provide novel insights for future endeavors in developing novel agents derived from natural product targeting CD73 enzyme.

Corticosteroid agonists of the glucocorticoid receptor are a mainstay of therapeutics for pro-inflammatory conditions. Vamorolone is a novel partial agonist that is differentiated from the other members of the corticosteroid class by non-metabolism by 11β-hydroxysteroid dehydrogenases, antagonism of the mineralocorticoid receptor, and differential co-factor binding. Our objective was to define the pharmacodynamic response of serum proteins to vamorolone.

Three ergostane-type steroids, including one rare C-29 methylated xylarstane A (1) and one C22,23-epoxied xylarstane B (2), along with one reported compound 3, were isolated from the endophytic fungus Xylaria sp. VDL4 harbored within the medicinal plant Vaccinium dunalianum. The structures of novel compounds were elucidated through comprehensive spectroscopic analysis, DP4-validated theoretical C nuclear magnetic resonance (NMR) calculations, and electronic circular dichroism (ECD) calculations. The antifungal activities of the isolates against four phytopathogens were assessed in vitro. Compounds 1 and 3 exerted significant inhibition against Alternaria solani and Botrytis cinerea respectively, both with minimal inhibitory concentration (MIC) of 12.5 μg/mL, comparing with the positive control (Carbendazim and Thiabendazole, MICs = 12.5-25.0 μg/mL).

Prostatic inflammation plays a pivotal role in prostate cancer development and progression via altering key cellular mechanisms, including proliferation, metastasis, and angiogenesis. Therefore, the use of anti-inflammatory drugs could provide a valid contribution to PCa prevention and treatment. In our research, we explored semi-synthetic derivatives of cycloastragenol (CA) and astragenol (AG) to assess their potential to inhibit inflammation-mediated tumorigenic signaling. Building on our previous findings, which demonstrated their inhibitory activity on NFκB, we discovered that these molecules also suppress inflammation-induced cell proliferation and migration through distinct mechanisms. They effectively alleviated inflammation by reducing levels of ROS, NO, and VEGF expression. Furthermore, these molecules partially restored the expression of AR and the tumor suppressor NKX3.1, both of which are critical in prostate tumorigenesis within an inflammatory microenvironment. They also reversed inflammation-induced activation of Akt and β-catenin signaling, suggesting their potential to inhibit inflammation-related prostate tumorigenesis. Our study further demonstrated that these molecules exhibited dose-dependent effects on inducing cell cycle arrest and apoptosis, as evidenced by increased p21 and decreased BCL-2 protein levels, leading to activated cell death and suppressed cellular migration. In conclusion, these semi-synthetic sapogenol derivatives demonstrate significant potential as anti-inflammatory and anticancer agents, offering a promising approach for targeting prostatic inflammation and inflammation-driven prostate carcinogenesis.

A fingernail clipping is expected to be a specimen suited for the detection of the medium-to-long term abnormal production of steroids. There are reports demonstrating that the concentration ratios of cortisol (F) to cortisone (E) and testosterone (T) to androstenedione (AD) in nails are significantly different from those in serum. However, the concentration ratios in the nails were determined with a small number of samples in some instances and differed from study to study, in addition to which, their gender differences remain poorly understood. Determining the differences in the concentration ratios between the nails and serum with a sufficient number of samples by gender and discussing a factor causing these differences will help for an accurate interpretation of the results in the nail-based steroid tests. In this study, we first developed and validated liquid chromatography/electrospray ionization-tandem mass spectrometry methods for determining the F/E and T/AD ratios in thumbnail clippings, then demonstrated that the F/E (in both sexes) and T/AD (in male) ratios in the nails were significantly lower than those in the serum reported in the literature. We found that steroids which are present at the higher concentrations in the nail than expected from their serum concentrations, such as E and AD, are bound to albumin at a high rate in the serum. As the affinities of corticoids and androgens to albumin are low (Ka, 10-10 M), we concluded that the albumin-bound fractions can be the sources of the nail-incorporated steroids as well as the free fractions.

Cardiovascular disease (CVD) and higher body mass index (BMI) are linked to chronic low-grade inflammation. It is unclear whether vitamin D modifies the association between CVD and inflammatory biomarkers uniformly across BMI strata. We therefore tested whether vitamin D status and supplementation modify the CVD-biomarker association and whether any modification differs by BMI category.

In contrast with previous results in which the spiroketal side chain of steroid sapogenin has proven to be unreactive to Jones reagent, treatment these sapogenins with silica gel-supported Jones reagent triggered the oxidative opening of the spirostanic side chain producing moderate to good yields of sapogenoic acids that bear carbonyl functions at C-16 and C-22 and a C-26 carboxylic group. Based on these findings, a procedure that allows the synthesis of sapogenoic acids minimizing contamination by chromium salts was designed.

Obesity-associated Dyslipidemia and inflammation are aggravated by the accumulation of toxic oxysterols, particularly 7-ketocholesterol (7-KC), which amplifies oxidative stress and metabolic dysfunction. The steroidal scaffold is a central determinant of whether such molecules exert protective or pathogenic effects. This review highlights the dichotomy between 7-KC, a cytotoxic oxysterol, and Guggulsterone, a Phytosteroids with therapeutic potential. Guggulsterone retains the tetracyclic steroid backbone, enabling it to mimic endogenous sterols while functioning as a farnesoid X receptor (FXR) antagonist. By relieving FXR-mediated suppression of CYP7A1, Guggulsterone enhances bile acid synthesis, promotes cholesterol clearance, and improves lipid profiles. Additionally, its structural features confer anti-inflammatory activity via NF-κB inhibition, contrasting with the pro-oxidant nature of 7-KC. The concept of steroid scaffold mimicry underscores the potential of Phytosteroids as blueprints for drug design, offering a path toward innovative therapies for obesity-linked metabolic disorders.

Breast cancer is the most common cancer in women, with triple-negative breast cancer (TNBC) accounting for approximately 20% of cases. TNBC lacks estrogen receptors (ER), progesterone receptors (PR), and epidermal growth factor receptor 2 (HER2) expression, which makes targeted therapies ineffective. The luminal androgen receptor (LAR) subtype of TNBC expresses androgen receptor (AR), highlighting the need for treatment strategies that target androgen signaling. Recently, the role of 11-oxygenated androgens, in addition to conventional androgens such as testosterone and dihydrotestosterone, in androgen-related diseases in women has gained increased attention. In this study, we investigated the involvement of 11-oxygenated androgens in LAR TNBC and explored the anti-androgenic effects of Kobochromone A (KC-A), a natural compound derived from Carex kobomugi. KC-A inhibits the androgen-synthesizing enzyme dehydrogenase/reductase short-chain dehydrogenase/reductase family member 11 (DHRS11) and suppresses AR expression. Using the AR-positive TNBC cell line MDA-MB-453, we demonstrated that 11-oxygenated androgens activate androgen signaling and promote cell proliferation. KC-A significantly inhibited androgen signaling by reducing nuclear AR localization and decreasing transmembrane protease, serine 2, and c-Myc expression. Furthermore, KC-A synergistically enhanced antiproliferative effects of the AKT inhibitor capivasertib (Cap), promoted apoptosis, and further suppressed AR expression. The primary therapeutic mechanisms of KC-A were identified as its dual actions: inhibition of DHRS11 and suppression of AR expression. These findings suggest that KC-A, either alone or in combination with AKT inhibitors, may offer a promising therapeutic strategy for LAR TNBC by targeting androgen signaling. Further studies are needed to confirm the efficacy and safety of KC-A in clinical applications.

The purpose of this study was to identify potential aromatase inhibitors, which might play a role in preventing breast cancer. The fungal-catalyzed microbial transformation of an anti-mineralocorticoid, canrenone (1), was catalyzed by Glomerella fusarioides, and Cunninghamella blakesleeana. Bioconversion of canrenone (1) with G. fusarioides provided a new polar metabolite 2, and two known metabolites 3 and 4, while C. blakesleana transformed compound 1 into two known polar metabolites 4, and 5. Modern spectroscopic techniques were employed to identify the structures of metabolites as 1-dehydro-11α-hydroxycanrenone (2), 1-dehydrocanrenone (3), 11α-hydroxycanrenone (4), and 11β-hydroxycanrenone (5). The SingleCrystal X-ray Diffraction (SCXRD) based structures of metabolites 2, and 3 are reported here for the first time. Canrenone (1) and the resulting metabolites 2-4 were evaluated for their human aromatase inhibitory activity. Compounds 1-4 showed the IC values of 0.288 ± 0.0392, 0.372 ± 0.002, 0.328 ± 0.0083, and 1.102 ± 0.099, µM comparable to the standard drug, exemestane (0.26 ± 0.011 µM). All transformed products were found non-cytotoxic to human fibroblast (BJ) cell line. Furthermore, the docking studies predicted the interaction of potential inhibitors with the active site residues of the enzyme via hydrogen bonding and other non-covalent interactions. Simulation studies predicted the formation of stable enzyme-inhibitor complexes with no or insignificant perturbation during the simulation time of 100 nsec. Hence, these inhibitors may serve as preliminary hits for drug discovery against ER+ breast cancer.

Nausea and vomiting in pregnancy (NVP) affect approximately 70 % of women worldwide. It is thought to have an adaptive function in the first trimester, when it protects the mother and the fetus against potential dangers from the diet. Proximate causes of NVP include hormonal changes during pregnancy. This longitudinal prospective study examined associations between various endogenous steroids and NVP. In the first and third trimester, pregnant women (N = 179) completed the Index of Nausea, Vomiting, and Retching questionnaire (92.1 % of women reported at least some symptoms of NVP in the first trimester and 37.4 % in the third trimester) and we analyzed their blood serum concentrations of 91 endogenous steroids. In the first trimester, NVP intensity was significantly positively associated with progesterone metabolites from the C21 5α/β-reduced steroid group (e.g., allopregnanolone sulfate) and with conjugated 5α-androstane-3α,17β-diol and conjugated 5α-androstane-3β,17β-diol. In the third trimester, we found significant negative associations between NVP and progesterone, conjugated testosterone, 7-oxo-DHEA, 5-androstene-3β,16α,17β-triol sulfate, some C21 Δ steroids (e.g., pregnenolone sulfate, 17-hydroxypregnenolone sulfate), and C21 5α/β-reduced steroids (such as allopregnanolone sulfate and conjugated pregnanolone). Our findings suggest that sulfated 3α-hydroxy-5α-steroids may contribute to NVP in early pregnancy by affecting brainstem regions involved in the vomiting reflex. In late pregnancy, higher levels of immunomodulatory androstanes and progesterone may reduce NVP severity via immune regulation and smooth muscle relaxation.