Antiandrogenic Activity Research Articles

Flutamide-induced alterations in transcriptional profiling of neonatal porcine ovaries

BackgroundAndrogens are involved in the regulation of ovarian development during fetal/neonatal life. Environmental chemicals displaying anti-androgenic activities may affect multiple signal transduction pathways by blocking endogenous androgen action. The aim of the current study was to examine effects of the anti-androgen flutamide on the expression of coding transcripts and long non-coding RNAs (lncRNAs) in neonatal porcine ovaries. By employing RNA-Seq technology we aimed to extend our understanding of the role of androgens in neonatal folliculogenesis and examine the impact of the anti-androgen flutamide on ovarian function.MethodPiglets were subcutaneously injected with flutamide (50 mg/kg BW) or corn oil (controls) between postnatal days 1 and 10 (n = 3/group). Ovaries were excised from the 11-day-old piglets and total cellular RNAs were isolated and sequenced.ResultsFlutamide-treated piglet ovaries showed 280 differentially expressed genes (DEGs; P-adjusted < 0.05 and log2 fold change ≥1.0) and 98 differentially expressed lncRNAs (DELs; P-adjusted < 0.05 and log2FC ≥ 1.0). The DEGs were assigned to GO term, covering biological processes, molecular functions and cellular components, which linked the DEGs to functions associated with cellular transport, cell divisions and cytoskeleton. In addition, STRING software demonstrated strongest interactions between genes related to cell proliferation. Correlations between DEGs and DELs were also found, revealing that a majority of the genes targeted by the flutamide-affected lncRNAs were associated with intracellular transport and cell division.ConclusionsOur results suggest that neonatal exposure of pigs to flutamide alters the expression of genes involved in ovarian cell proliferation, ovarian steroidogenesis and oocyte fertilization, which in turn may affect female reproduction in adult life.

Comparative transcriptional analysis of methylparaben and propylparaben in zebrafish

Parabens are widely used as preservatives in different commercial items including food, cosmetics and pharmaceuticals, and their wide use has resulted in accumulation in the environment. Parabens have been shown to have negative effects on animals as well as human health. In this study, we carried out a comprehensive study to determine the adverse effects associated with propylparaben (PP) and methylparaben (MP) on early developmental stages of zebrafish. Mortality, hatching, developmental abnormalities and gene expression profiles were investigated in embryos exposed to both compounds. The semi-static exposure conditions showed that both MP (≥100 μM) and PP (≥10 μM) are toxic to the embryos in a concentration-dependent manner and lead to developmental abnormality. Malformations such as spinal defects, pericardial edema, and pigmentation defects were observed following both MP and PP treatments. Hatching delay, mortality and developmental abnormality data indicate that PP is more toxic than MP. For gene expression analysis, 1 and 10 μM doses of MP and PP were analyzed. Genes from physiological pathways including stress response, cell cycle and DNA damage, inflammation, fatty acid metabolism and endocrine functions were affected by MP and PP. The gene expression profiles show that parabens cause toxicity by inducing oxidative stress, DNA double-strand breaks, apoptosis as well as by altering fatty acid metabolism. Altered expression of androgen receptor (ar) and estrogen receptor 2 alpha (esr2a) indicates an antiandrogenic and estrogenic activity of parabens in zebrafish. Overall, the present study provides considerable information on the negative effects of MP and PP using physiological endpoints and motivates further studies to explore the molecular mechanism of the toxicity associated with parabens.

Ethanolic extract of Algerian propolis decreases androgen receptor transcriptional activity in cultured LNCaP cells

Antiandrogens have a peculiar place in the treatment of metastatic prostate cancer by blocking the androgen receptor (AR). Unfortunately, aggressive tumors could rapidly develop into a castration resistant state. It is therefore essential to look for new molecules that are more effective, affecting not only the androgen signaling and with minimum undesirable effects. Natural products are an interesting source of new therapeutics, especially for cancer therapy as 70% of them have botanical origin. Based on an ethnobotany screening, we evaluated the effects of ethanolic extract of propolis (EEP) from Algeria on LNCaP cells. Results pointed out that EEP reduces the survival of LNCaP cells with an IC50 of 0.04 mg/ml, induces the apoptosis and blocks the cell cycle at G0/G1 phase. Interestingly, EEP decreased the accumulation of AR suggesting some anti-androgen activity. Indeed, secreted amount of the androgen target protein PSA was decreased when LNCaP cells were incubated with EEP, starting after 4 h of treatment. This anti-androgen activity was also shown on the androgen target genes Fkbp5 and Sgk1. Finally, the capacity of EEP to block AR functioning was demonstrated in transient transfections with human AR and the reporter gene ARE-tk-Luc. Propolis antagonizes the induction of the luciferase activity induced by the natural androgen DHT (10−8M) or the synthetic AR agonist R1881 (10-7M). Altogether, these results highlight the potential pharmacological effects of EEP in future treatments of prostate cancer.

Study of gonado- and reproductive toxicity of technical-grade carbendazim in male and female Wistar Han rats

Objective. Identification of hazard and assessment of the risk of gonado- and reproductive toxicity of two generic pesticides of technical-grade carbendazim of different purity bin male and female Wistar Han rats. Materials and Methods. Carbendazim with the purity of 98 % (C1) and 98.1 % (C2) was intragastrically administered on the daily basis, excluding Saturday and Sunday, at the doses of 2.5 and 25 mg/kg body weight in three groups of animals (20 males and female rats in each) during 10 weeks (forfemales) and 11 weeks (for males). Control animals received an equivalent amount of the solvent. Intact animals intended for breeding were managed in parallel with the control and experimental animals. After the end of the exposure, functional parameters of the condition of gonads and reproductive ability of the animals were investigated. Females were used to investigate the condition of the oestral cycle, duration and frequency of each stage of it. The condition of the reproductive function in the female rats was considered at Day 20 of pregnancy. Indices of breeding, conception, fertility, pregnancy were established, and the duration of the pre-coitalperiod was considered. Results and Discussion. The data obtained suggest that exposure to two generic substances of carbendazim at the dose of 25 mg/kg body weight during 11 weeks is manifested in general toxic effect (body weight reduction) only in males upon exposure to C2. All test substances in the maximum dose have reproductive toxicity and show significant anti-androgenic action that is manifested as changes in the morphological and functional parameters of the condition of gonads in the experimental groups of male rats. Furthermore, the negative effect on the oestral cycle in experimental female rats exposed to C2 was noted. Conclusion. Based on the obtained results it can be concluded that all test samples of carbendazim have reproductive toxicity at the dose of 25 mg/kg body weight. No-observed-effect level (NOEL) for all test compounds is 2.5 mg/kg body weight. Dose-effect dependence was observed in the range of studies doses.

Abstract OT1-02-01: Phase I trial of bicalutamide and ribociclib in androgen receptor-positive triple negative breast cancer

Abstract Background: To date, the use of anti-androgens in the subset of triple negative breast cancers (TNBC) that express androgen receptor (AR) has shown modest response rates, indicating anti-androgen-resistance in the majority of these tumors. Based on data that Cyclin D kinase (CDK) inhibitors reverse resistance to anti-androgens in prostate cancer cell lines, we hypothesize that the use of CDK inhibition may enhance the activity of anti-androgens in AR-positive TNBC. Methods: Key eligibility include: patients with centrally confirmed AR-positive TNBC, defined as AR expression &amp;gt;0%; 0 to 1 line of prior therapy for metastatic disease; and measurable disease. Patients are treated with bicalutamide 150mg orally once daily plus ribociclib at one of 3 dose levels (see table). Table 1Dose levelBicalutamideRibociclib1150mg orally once daily (D1 to 28)400mg daily (D1 to 21)2150mg orally once daily (D1 to 28)400mg daily (D1 to 28)3150mg orally once daily (D1 to 28)600mg daily (D1 to 21) Results: AR expression was positive by trial criteria in 74% of screened patients. Three patients have been accrued at each dose level. Median age is 56 and 6 and 3 patients were treated in first and second-line settings, respectively. Median AR expression was 50% (range 5 to 75%). Toxicity data is available for 6-patients treated on dose levels 1 and 2. No dose-limiting toxicities were noted. As anticipated with ribociclib, the most common toxicity is neutropenia (1 patient grade 4 and 2 patients grade 3). Two patients experienced grade 3 hypertension and 1 experienced grade 3 lymphopenia. Grade 2 or lower toxicities included fatigue, nausea, hyperglycemia and mucositis. One patient experienced grade 1 QT interval prolongation. Conclusion: The combination of bicalutamide and ribociclib is tolerable without unexpected toxicities. Data on the 3-patients treated at dose level 3 and dose expansion will be included. Phase 2 dosing schedule will be decided based on phase 1 results. Citation Format: Sharifi M, Wisinski KB, Burkard ME, Tevaarwerk AJ, Tamkus D, Chan N, Truica C, Danciu O, Hoskins K, O'Regan RM. Phase I trial of bicalutamide and ribociclib in androgen receptor-positive triple negative breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr OT1-02-01.

Biotransformation into 11α-hydroxyprogesterone glucosides by glucosyltransferase

Recently, studies on the steroidal hormone activity in the brain have attracted attention, and the influences of the varied glucosides and their artificial derivatives have been discussed; additionally, it has been suggested that glucosides are the synthetic precursors of glucuronide as a label molecule. However, glucosides are formed with 11α-hydroxyprogesterone (1), which is important as a blood pressure regulator, but anti-androgen activity remains unknown. Using UDP-glucosyltransferase, glucoside synthesis was successful in linking β-d-glucopyranose and β-d-laminaribiose to 11α oxygen of 1 at a high conversion ratio, and full assignment structure was analyzed for the two glucosides by high-resolution quadrupole-time flight electrospray ionization-mass spectrometry, 1D (1H and 13C) NMR and 2D (COSY, ROESY, HSQC-DEPT and HMQC) NMR. Furthermore, the bioactivity of 1 and two 11α-hydroxyprogesterone glucosides [11α-(β-d-glucopyranosyl)oxyprogesterone, 2, and 11α-(β-d-laminaribiosyl)oxyprogesterone, 3] was tested in vitro. On rotenone-induced PC12 cells, the two 11α-hydroxyprogesterone glucosides (2 and 3) showed superior neuroprotective effects and increased cellular ATP levels compared with those of 1.

Assessment of endocrine-disrupting activities of alternative chemicals for bis(2-ethylhexyl)phthalate

Plastic products are closely intertwined with modern life. Some plasticizers used in making plastics, such as phthalates, are reported to be endocrine-disrupting chemicals. Plasticizers can be released into the environment, and health risks related to plasticizer exposure have been reported. In addition, due to plastic waste that flows into the ocean, microplastics have been found in marine products, including non-biological seawater products such as sea salt. Plastics can affect the body via a variety of pathways, and therefore safer alternative chemicals are needed. Three chemicals were evaluated: acetyl tributyl citrate (ATBC), triethyl 2-acetylcitrate (ATEC), and trihexyl O-acetylacitrate (ATHC), replacing bis(2-ethylhexyl)phthalate (DEHP), a typical plasticizer. The endocrine-disrupting activities of each chemical, including estrogenic or anti-estrogenic activity (test guideline (TG) No. 455), androgenic or anti-androgenic activity (TG No. 458), steroidogenesis (TG No. 456), and estrogenic properties via a short-term screening test using the uterotrophic assay (TG No. 440), were assessed in accordance with the Organisation for Economic Co-operation and Development guidelines for chemical testing. Our results showed that DEHP, ATBC, ATEC, ATHC possess no estrogenic activity, whereas DEHP, ATBC and ATHC demonstrate anti-estrogenic activity and ATBC anti-androgenic activity. DEHP and ATHC exhibited a disruption in steroidogenesis activities. Additional tests are necessary, but our results suggest that ATEC is a good candidate plasticizer providing a suitable alternative to DEHP.

Autism spectrum disorders, endocrine disrupting compounds, and heavy metals in amniotic fluid: a case-control study

BackgroundEvidence has indicated that some non-inherited factors such as exposure to environmental pollutants are associated with neurodevelopment disorders like autism spectrum disorder (ASD). Studies report that endocrine disrupting compounds (EDCs), including polychlorinated biphenyls, organochlorine pesticides, perfluoroalkyl substances (PFAS), and some metals, have adverse effects on the fetal neurodevelopment. The aim of this study was to measure the amniotic fluid (AF) levels of EDCs and metals as well as the receptor transactivities induced by AF and investigate the possible link between prenatal exposure to EDCs and heavy metals and ASD risk.MethodsIn this case-control study, we included AF samples of 75 ASD cases and 135 frequency-matched controls and measured the levels of the endogenous sex hormones, PFAS, and elements including heavy metals. The combined effect of endogenous hormones and EDCs on the receptor of estrogen (ER), androgen (AR), aryl hydrocarbon (AhR), and thyroid hormone-like activity were also determined and expressed as receptor ligand equivalents. We assessed the associations of AF levels of chemicals, sex hormones, and receptor activities with ASD risk using unconditional logistical regression analyses. To control for multiple comparisons, the false discovery rate (FDR) was used and q values less than 0.25 were designated as statistical significance.ResultsPFAS and metals were detectable in AF samples. The ASD cases had significantly lower AF levels of PFAS than controls, and the adjusted odds ratio (OR) was 0.410 (95% CI 0.174, 0.967; p = 0.042; FDR qvalue = 0.437) for perfluorooctane sulfonate (PFOS). The principal component, including PFAS congeners, copper, iron, and estrogenic activity, was significantly inversely associated with ASD risk (adjusted OR = 0.100; 95% CI 0.016, 0.630; p = 0.014; FDR qvalue = 0.098).Testosterone level in AF weakly associated with ASD risk (adjusted OR = 1.002; 95% CI 1.000, 1.004; p = 0.05). However, after multiple comparison correction, the association was not significant (FDR qvalue = 0.437). No significant associations between AF-induced receptor transactivities and ASD risk were observed. The adjusted OR was 2.176 (95%CI 0.115, 41.153) for the ratio of the combined androgenic activity to combined estrogenic activity.ConclusionsThe presence of PFAS and heavy metals in AF indicates that they can cross the placenta. The inverse association between levels of PFAS congeners in AF and ASD risk might relate to the weak estrogenic activities and anti-androgenic activities of PFAS.The observed tendency of positive association between the ratio of combined androgenic effect to the combined estrogenic effect and ASD risk needs further studies to explore whether EDCs together with endogenous hormones play a role in the development of ASD.

Anti-androgenic and insulin-sensitizing actions of Nigella sativa oil improve polycystic ovary and associated dyslipidemia and redox disturbances

Aim: Nigella sativa oil has been shown to improve metabolic syndrome-associated dyslipidemia and oxidative stress. Effects of N. sativa oil on letrozole-induced polycystic ovary syndrome (PCOS)-associated hormonal, metabolic, redox, and lipid dysregulation in rats were investigated. Methods: Female Sprague-Dawley rats were randomly assigned into four groups (n=10 per group): control, PCOS, and PCOS treated with N. sativa oil (5 mL/kg/day or 10 mL/kg/day) once daily p.o. starting at day 7 after the commencement of letrozole (1mg/kg) until day 56. Body weight and estrous cycle were recorded starting from days 7 and 28 respectively until day 56. Rats were dissected and serum and ovaries were collected for biochemical and histomorphological assessments. Results: N. sativa oil (10 mL/kg/day) treatment significantly increased: number of rats undergoing regular cycle (80% vs 20%; p< 0.05), average number of regular cycles (5.00±0.59 vs 0.20±0.42; p< 0.05), appearance of corpus luteum; but reduced number of cystic follicles (7.60±1.51 vs 6.10±0.19; p< 0.05) in PCOS rats. Body weight was also reduced by the oil in PCOS rats. However, ovarian weight, granulose and theca cells layers showed no significant change. The oil also restored altered circulating levels of gonadotropins, steroids and reduced fasting blood glucose. The ovarian total cholesterol, triglycerides, low density lipoprotein-cholesterol, malondialdehyde were markedly reduced while insulin sensitivity and ovarian high density lipoprotein-cholesterol, superoxide dismutase and glutathione peroxidase activities were greatly increased. Conclusion: N. sativa oil improved polycystic ovarian-morphology and metabolic disorders via its anti-androgenic and insulin-sensitizing actions to ameliorate PCOS-associated dyslipidemia and redox disturbances.

The effects on the endocrine system under hepatotoxicity induction by phenobarbital and di(2-ethylhexyl)phthalate in intact juvenile male rats

Phenobarbital (PB) and Di (2-ethylhexyl) phthalate (DEHP), an anti-epileptic drug and a plasticizer used in flexible polyvinylchloride formulations, respectively, are well-known typical hepatotoxicants. This study investigated the effects of PB (100 mg/kg/day) or DEHP (500 mg/kg/day) on the endocrine system in intact juvenile/peripubertal male rats exposed for 31 days beginning on postnatal day 23. Slight hormone level changes, histopathological changes in thyroid gland or induction of UDP-glucuronosyltransferase in liver were observed in both the PB and DEHP groups. One of the assumed mechanisms inducing thyroid effects is predictable to be secondary changes based on the enhancement in thyroid hormone metabolism via the induction of hepatic microsomal enzymes. No reproductive system-related changes in organ weights, histopathology, and sexual maturation were observed in both groups. Lower testosterone level was observed in the PB group. CYP2B and CYP3A, which are involved in testosterone metabolism, were induced in liver of the PB group. There was no change of 17β-hydroxysteroid dehydrogenase activity in testis of both groups. Lower testosterone level in the PB-treated male rats was attributed to an indirect, hepatotoxicity-associated effect on the reproductive system and not to direct effects on testis such as the antiandrogenic activity and the inhibition of steroidogenesis. These results did not indicate that PB or DEHP exposure affects the endocrine system directly.

Protective effect of flavonoids from Foeniculum vulgare against ultraviolet-B-induced oxidative stress in human dermal fibroblasts

Background: Traditionally, Foeniculum vulgare (fennel seeds) has been used for its antimicrobial, analgesic, antipyretic, antiflatulence, antispasmodic, and antiandrogenic activities. Materials and Methods: In the present study, the protective effect of flavonoids from fennel seeds was investigated against ultraviolet (UV)-B radiation-induced cell damage and oxidative stress in human dermal fibroblast (HDF) cells. Results: Flavonoid-enriched fraction (FEF) of fennel seeds showed high flavonoid content and antioxidant potential as well as the presence of a marker compound rutin. Pretreatment of HDF cells with the FEF (15–45 μg/ml) significantly protected against UV-B-induced cytotoxicity, endogenous enzymatic antioxidant depletion, oxidative DNA damage, intracellular reactive oxygen species generation, and apoptotic morphological changes. Conclusion: The current study proved for the first time that the FEF of fennel seeds reduced oxidative stress through the nuclear factor E2-related factor 2-antioxidant response elements pathway. Flavonoids from fennel seeds have a potential as UV-B protectants and can be explored against diseases, in which oxidative stress is closely implicated.

Determination of bisphenol A and bisphenol S concentrations and assessment of estrogen- and anti-androgen-like activities in thermal paper receipts from Brazil, France, and Spain

Bisphenol A (BPA) is a high-production-volume chemical with endocrine disrupting properties commonly used as color developer in thermal paper. Concerns about the potential hazards of human BPA exposure have led to the increasing utilization of alternatives such as bisphenol S (BPS) and bisphenol F (BPF). This study was designed to assess: (i) BPA, BPS, and BPF concentrations in 112 thermal paper receipts from Brazil, France, and Spain by liquid chromatography coupled to mass spectrometry (LC-MS); and (ii) hormone-like activities of these receipts using two receptor-specific bioassays, the E-Screen for (anti-)estrogenicity and PALM luciferase assay for (anti-)androgenicity. BPA was present in 95.3% of receipts from Spain, 90.9% of those from Brazil, and 51.1% of those from France at concentrations up to 20.27 mg/g of paper. Only two samples from Brazil, two from Spain, and ten from France had a BPS concentration ranging from 6.46 to 13.29 mg/g; no BPA or BPS was detected in 27.7% of French samples. No BPF was detected in any receipt. Estrogenic activity was observed in all samples from Brazil and Spain and in 74.5% of those from France. Anti-androgenic activity was observed in > 90% of samples from Brazil and Spain and in 53.2% of those from France. Only 25.5% of French samples were negative for both estrogenic and anti-androgenic activity. Estrogenic and anti-androgenic activities per gram of paper were up to 1.411 µM estradiol (E2) equivalent units (E2eq) and up to 359.5 mM procymidone equivalent units (Proceq), respectively. BPA but not BPS concentrations were positively correlated with both estrogenic and anti-androgenic activities. BPA still dominates the thermal paper market in Brazil and Spain, and BPS appears to be one of the main alternatives in France. There is an urgent need to evaluate the safety of alternatives proposed to replace BPA as developer in thermal printing. The large proportion of samples with hormonal activity calls for the adoption of preventive measures.

Effects on tadpole snail gene expression after exposure to vinclozolin

The fungicide vinclozolin (Vz) is an endocrine disruptor with known anti-androgenic activity in vertebrates. However, there is a lack of information about the Vz mode of action in invertebrates, although some studies have shown that this compound can produce alterations in different species. Transcriptional activity was analyzed in the freshwater snail Physella acuta in order to elucidate putative cellular processes altered by this chemical during a response. In order to identify potential molecular biomarkers, a de novo transcriptome was generated for this species that constitutes a valuable source for future studies. This data, together with some already available data, permitted the identification of several genes related to detoxification mechanisms (Cyp2u1, Cyp3a7, Cyp4f22, GSTo1, GSTt2, and MRP1), stress response (Hsp20.4, Hsp17, Hsp16.6, and Cu,Zn-SOD), the hormonal system (Estrogen Receptor and Hsp90), apoptosis (Casp3), and copper homeostasis (ATOX1). Using quantitative Real-Time polymerase chain reaction, mRNA levels of these genes were examined in snails exposed to 20 or 200 µg/L Vz for 24 h. The results showed an overall weak response, with downregulation of Hsp20.4 and no statistically significant change for the other genes. These findings suggest that P. acuta can manage the concentrations of Vz found in the environment with no relevant activation of the pathways analyzed, although additional studies are needed for longer exposure times and including other metabolic pathways. The new genes described open the range of processes that can be studied at the molecular level in toxicity tests.

Therapeutic tactics with violations of the menstrual cycle caused by ovulative dysfunction

Purpose: presentation of modern data on the pathogenesis of menstrual cycle disorders (NMCs) associated with ovulatory dysfunction, and the effectiveness of their treatment with medication.Basic provisions. It is shown that NMC is the result of violations of the cyclic production of gonadotropic and sex hormones. Among the hormonal drugs for the treatment of NMCs, combined estrogen-progestational oral contraceptives (COCs), especially those with drospirenone (DRSP), are advantageous. The introduction of calcium levomefolate - biologically active form of folatein the composition of COCs containing 30 μg of ethinylestradiol and DRSP has a potentiating effect associated with their noncontraceptive effects due to antiandrogenic and antimineralocorticoid activity. It was shown that calcium levomefolate, which participates in human body metabolism, as a component of COC, has a protective effect on the cardiovascular system (due to a decrease in the level of homocysteinemia) in the syndrome of polycystic ovaries, contributes to reducing obesity.Conclusion. The combination of 30EE/DRSP promotes the normalization of the menstrual cycle and has non-contraceptive effects associated with antiandrogenic and antimineralcorticoid action, and fortification with calcium levomefolate also has a positive effect on the hormonal profile and lipid metabolism, has a protective effect on cardiovascular diseases in women with the metabolic syndrome in polycystic ovaries, contributes to the normalization of weight.

Employing Dietary Comparators to Perform Risk Assessments for Anti-Androgens Without Using Animal Data.

This study investigated the use of androgen receptor (AR) reporter gene assay data in a non-animal exposure-led risk assessment in which in vitro anti-androgenic activity and exposure data were put into context using a naturally occurring comparator substance with a history of dietary consumption. First, several dietary components were screened to identify which selectively interfered with AR signaling in vitro, using the AR CALUX® test. The IC50 values from these dose-response data together with measured or predicted human exposure levels were used to calculate exposure: activity ratios (EARs) for the dietary components and a number of other well-known anti-androgenic substances. Both diindolylmethane (DIM) and resveratrol are specifically acting dietary anti-androgens. The EARs for several anti-androgens were therefore expressed relative to the EAR of DIM, and how this ‘dietary comparator ratio’ (DCR) approach may be used to make safety decisions was assessed using an exposure-led case study for an anti-androgenic botanical ingredient. This highlights a pragmatic approach which allows novel chemical exposures to be put into context against dietary exposures to natural anti-androgenic substances. The DCR approach may have utility for other modes of action where appropriate comparators can be identified.

Effect-based and chemical analyses of agonistic and antagonistic endocrine disruptors in multiple matrices of eutrophic freshwaters

In the present study, both bioanalytical and instrumental tools were employed to examine the endocrine-disruptive potentials of water samples, cyanobloom samples, and sediment samples collected from in the northern region of Taihu Lake (China) during cyanobloom season. Results from cell-based bioassays suggested the occurrence of estrogenic, anti-estrogenic, anti-androgenic, and anti-glucocorticogenic activities, while no androgenic and glucocorticogenic activities were observed in the collected samples. Using an UPLC-MS/MS system, 29 endocrine disrupting compounds including seven estrogens, seven androgens, six progestogens, and five adrenocortical hormones and four industrial pollutants were simultaneously detected. 17, 20 and 12 chemicals were detected at least in one of the water samples, cyanobloom samples and sediment samples, respectively. Since both agonistic and antagonistic endocrine-disruptive activities were detected in the present study, commonly used receptor-based in vitro bioassays resulted in net effects, suggesting that the hormone receptor agonistic potentials might be underestimated with this practice. The EDCs detected in cyanobloom samples also highlight the necessity to consider the phytoplankton matrix for understanding the mass fluxes of endocrine disruptors in eutrophic freshwaters and to consider it in monitoring strategies.

Novel Trifluoromethylated Enobosarm Analogues with Potent Antiandrogenic Activity In Vitro and Tissue Selectivity In Vivo

Prostate cancer often develops antiandrogen resistance, possibly via androgen receptor (AR) mutations, which change antagonists to agonists. Novel therapies with increased anticancer activity, while overcoming current drug resistance are urgently needed. Enobosarm has anabolic effects on muscle and bone while having no effect on the prostate. Here, we describe the activity of novel chemically modified enobosarm analogues. The rational addition of bis-trifluoromethyl groups into ring B of enobosarm, profoundly modified their activity, pharmacokinetic and tissue distribution profiles. These chemical structural modifications resulted in an improved AR binding affinity-by increasing the molecular occupational volume near helix 12 of AR. In vitro, the analogues SK33 and SK51 showed very potent antiandrogenic activity, monitored using LNCaP/AR-Luciferase cells where growth, PSA and luciferase activity were used as AR activity measurements. These compounds were 10-fold more potent than bicalutamide and 100-fold more potent than enobosarm within the LNCaP model. These compounds were also active in LNCaP/BicR cells with acquired bicalutamide resistance. In vivo, using the AR-Luc reporter mice, these drugs showed potent AR inhibitory activity in the prostate and other AR-expressing tissues, e.g., testes, seminal vesicles, and brain. These compounds do not inhibit AR activity in the skeletal muscle, and spleen, thus indicating a selective tissue inhibitory profile. These compounds were also active in vivo in the Pb-Pten deletion model. SK33 and SK51 have significantly different and enhanced activity profiles compared with enobosarm and are ideal candidates for further development for prostate cancer therapy with potentially fewer side effects. Mol Cancer Ther; 17(9); 1846-58. ©2018 AACR.

Development of emulgel technology with saw palmetto extracts and of japanese sophora tincture for androgenetic alopecia treatment

Androgenic alopecia is a hair disease caused by an excessive influence of androgens on hair follicles, whose treatment requires long-term use of systemic and / or local action drugs. In Ukrainian pharmaceutical market for the androgenic alopecia treatment synthetic drug minoxidil is presented, which in the local long-term application causes many side effects. Therefore, the drugs development of plant origin is important.&#x0D; Taking into account of the androgenic alopecia pathogenesis, it was developed the emulgel composition and technology, which has Saw Palmetto extract and Sophora Japonica tincture as active ingredients. Their main biological active substances applying topically provide anti-androgenic, venotonic and follicles protective action.&#x0D; The qualities of dermatological remedies in large measure are influenced by their technological process. Therefore, the purpose of this investigation was the development of a rational emulgel technology with Saw Palmetto extract and Sophora Japonica tincture for the androgenic alopecia prevention and treatment.&#x0D; Samples of emulgel were manufactured with applying of different technological methods, namely: the method of emulsification, temperature, rate and duration of stirring. Prepared samples were controlled by uniformity, dispersion degree (with microscope Delta Optical Genetic Pro, Poland), pH (with pH-meter 150 MI, Russia) and structure viscosity (with rotational viscometer Brookfield VR-3000, Myr VISKOTECH, Spain).&#x0D; The optimal temperature for the emulgel preparation is 60 °C, since the melting temperature of cetyl alcohol (the most refractory component of the formulation) is 52 °C. When choosing the emulsifying method it was found that sequential addition of hydrophilic and oil phases to emulgators provided the highest dispersion and optimally rheological properties of the emulgel.&#x0D; The best homogenization speed, providing a high degree of dispersion and structural viscosity in the range 3 200–3 400 mPas, is 2 000 rev/min. The intensive size reduction of the dispersed phase was observed for 30 min stirring.&#x0D; As a result of investigation, it was established that the best method of emulsification was the sequential adding of oil and aqueous phase to the mixture of emulsifiers at the temperature – 60 °C, the speed – 2 000 rev/min for 30 min.

A REVIEW ON ANTIFERTILITY ACTIVITY OF PLANTS OF CHAMBAL RIVERINE

Indian sub-continent showing very high diversity in both ecological and geographical aspects. In India, medicinal plants are widely distributed in different climatic conditions and are abundant in a mountainous region or riverine area. In this review, are being discussed anti-fertility properties of India’s well-known Chambal riverine flora. This area is rich in many medicinal plants as Acyranthus aspera, Acacia catechu, Nelumbo nucifera, Nyctanthes arbortristis, Mimosa pudica, Adhatoda vasica, Calotropis gigantea, Pergularia daemia, Vicoa indica, etc. These plants possess potential antifertility activity and many phytoconstituents, responsible for antispermatogenic, anti-implantation, antiovulatory, antiandrogenic, and many more activities. Hence, this review comprises a brief study of some medicinal plants found in Chambal riverine and showing the comprehensive antifertility activity of those plants.

Unexpected, ubiquitous exposure of pregnant Brazilian women to diisopentyl phthalate, one of the most potent antiandrogenic phthalates

BackgroundHuman exposure to phthalates and other non-persistent chemicals in developing countries is largely unknown. A preliminary analysis of urinary samples from pregnant Brazilian women revealed the presence of metabolites of Diisopentyl phthalate (DiPeP). ObjectivesReliably quantify DiPeP metabolites in human urine and investigate the potential antiandrogenic activity of this phthalate in rats. MethodsWe initiated a pilot pregnancy cohort in Curitiba, Brazil, to examine phthalate exposure in urine samples collected in early pregnancy (n = 50) or pooled samples from early, mid and late pregnancy (n = 44). Our well established phthalate method was modified to include the primary DiPeP metabolite, monoisopentyl phthalate (MiPeP), and two additional secondary oxidized metabolites, 3OH-MiPeP and 4OH-MiPeP. In a parallel approach, we orally exposed pregnant rats to DiPeP or Di-n-butyl phthalate (DnBP; reference phthalate) at 0, 125, 250, and 500 mg/kg/day from gestation day 14 to 18 and measured ex vivo fetal testis testosterone production. ResultsWe were able to detect and quantify specific DiPeP metabolites in nearly all (98%) of the early pregnancy urine samples and in all gestational pool samples with a median concentration for MiPeP of 3.65 and 3.15 μg/L, respectively, and for the two oxidized metabolites between 1.00 and 1.70 μg/L. All three urinary DiPeP metabolites were strongly correlated (r = 0.89 to 0.99). In the rat model, the effective dose (mg/kg/day) inhibiting fetal testosterone production by 50% (ED50 [95% confidence interval]) was 93.6 [62.9–139.3] for DiPeP which was significantly lower than for DnBP (220.3 [172.9–280.7]), highlighting the strong antiandrogenic potency of DiPeP within the spectrum of the phthalates. ConclusionsWe unveiled and confirmed the exposure of pregnant Brazilian women to DiPeP via specific urinary metabolites. This unexpected and ubiquitous DiPeP exposure indicates to unique DiPeP exposure sources in Brazil. These exposures spark considerable concern because DiPeP is one of the most potent antiandrogenic phthalates.