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The Journal of Steroid Biochemistry and... Jul 2023Dysfunction of the androgen receptor (AR) signalling axis plays a pivotal role in the development and progression of prostate cancer (PCa). Steroidal and non-steroidal...
Dihydrotestosterone-based A-ring-fused pyridines: Microwave-assisted synthesis and biological evaluation in prostate cancer cells compared to structurally related quinolines.
Dysfunction of the androgen receptor (AR) signalling axis plays a pivotal role in the development and progression of prostate cancer (PCa). Steroidal and non-steroidal AR antagonists can significantly improve the survival of PCa patients by blocking the action of the endogenous ligand through binding to the hormone receptor and preventing its activation. Herein, we report two synthetic strategies, each utilizing the advantages of microwave irradiation, to modify the A-ring of natural androgen 5α-dihydrotestosterone (DHT) with pyridine scaffolds. Treatment of DHT with appropriate Mannich salts led to 1,5-diketones, which were then converted with hydroxylamine to A-ring-fused 6'-substituted pyridines. To extend the compound library with 4',6'-disubstituted analogues, 2-arylidene derivatives of DHT were subjected to ring closure reactions according to the Kröhnke's pyridine synthesis. The crystal structure of a monosubstituted pyridine product was determined by single crystal X-ray diffraction. AR transcriptional activity in a reporter cell line was investigated for all novel A-ring-fused pyridines and a number of previously synthesized DHT-based quinolines were included to the biological study to obtain information about the structure-activity relationship. It was shown that several A-ring-fused quinolines acted as AR antagonists, in comparison with the dual or agonist character of the majority of A-ring-fused pyridines. Derivative 1d (A-ring-fused 6'-methoxyquinoline) was studied in detail and showed to be a low-micromolar AR antagonist (IC = 10.5 µM), and it suppressed the viability and proliferation of AR-positive PCa cell lines. Moreover, the candidate compound blocked the AR downstream signalling, induced moderate cell-cycle arrest and showed to bind recombinant AR and to target AR in cells. The binding mode and crucial interactions were described using molecular modelling.
Topics: Male; Humans; Dihydrotestosterone; Microwaves; Receptors, Androgen; Prostatic Neoplasms; Cell Line, Tumor; Androgen Receptor Antagonists; Pyridines
PubMed: 37086925
DOI: 10.1016/j.jsbmb.2023.106315 -
Journal of Veterinary Internal Medicine Nov 2021The use of adrenocorticotropic hormone stimulation test as method to monitor efficacy of trilostane treatment of hypercortisolism (HC) in dogs has been questioned.
BACKGROUND
The use of adrenocorticotropic hormone stimulation test as method to monitor efficacy of trilostane treatment of hypercortisolism (HC) in dogs has been questioned.
OBJECTIVES
To evaluate and compare 12 methods with which to monitor efficacy of trilostane treatment in dogs with HC.
ANIMALS
Forty-five client-owned dogs with HC treated with trilostane q12h.
METHODS
Prospective cross-sectional observational study. The dogs were categorized as well-controlled, undercontrolled, and unwell through a clinical score obtained from an owner questionnaire. The ability to correctly identify trilostane-treatment control of dogs with HC with the following variables was evaluated: before trilostane serum cortisol (prepill), before-ACTH serum cortisol, post-ACTH serum cortisol, plasma endogenous ACTH concentrations, prepill/eACTH ratio, serum haptoglobin (Hp) concentration, serum alanine aminotransferase (ALT), gamma-glutamyl transferase (γGT) and alkaline phosphatase activity, urine specific gravity, and urinary cortisol : creatinine ratio.
RESULTS
Ninety-four re-evaluations of 44 dogs were included; 5 re-evaluations of 5 unwell dogs were excluded. Haptoglobin was significantly associated with the clinical score (P < .001) and in the receiver operating characteristic analysis, Hp cutoff of 151 mg/dL correctly identified 90.0% of well-controlled dogs (specificity) and 65.6% of undercontrolled dogs (sensitivity). Alanine aminotransferase (P = .01) and γGT (P = .009) were significantly higher in undercontrolled dogs. Cutoff of ALT and γGT greater than or equal to 86 U/L and 5.8 U/L, respectively, were significantly associated with poor control of HC by trilostane.
CONCLUSIONS AND CLINICAL IMPORTANCE
Of all the 12 variables, Hp, and to a lesser degree ALT and γGT, could be considered additional tools to the clinical picture to identify well-controlled and undercontrolled trilostane-treated dogs.
Topics: Adrenocortical Hyperfunction; Animals; Cross-Sectional Studies; Cushing Syndrome; Dihydrotestosterone; Dog Diseases; Dogs; Enzyme Inhibitors; Hydrocortisone; Prospective Studies
PubMed: 34672018
DOI: 10.1111/jvim.16269 -
International Journal of Sports... Nov 2021To establish if training volume was associated with androgen baselines and androgen responsiveness to acute exercise.
PURPOSE
To establish if training volume was associated with androgen baselines and androgen responsiveness to acute exercise.
METHODS
During a "high-volume" training phase, 28 cyclists (14 men and 14 women) undertook oxygen-uptake and maximal-work-capacity testing. Two days later, they completed a repeat-sprint protocol, which was repeated 3 weeks later during a "low-volume" phase. Blood and saliva samples were collected before and after (+5 and +60 min) the repeat-sprint protocol. Blood was assayed for total testosterone (TT), free testosterone (FT), and dihydrotestosterone (DHT) and saliva, for testosterone and DHT.
RESULTS
Pretrial TT, FT, and DHT concentration was greater for males (P < .001, large effect size differences), and in both genders TT, DHT, and saliva for DHT was higher during high-volume loading (moderate to large effect size). Area-under-the-curve analysis revealed larger TT, FT, and DHT responses to the repeat-sprint protocol among females, and high-volume training was linked to larger TT, DHT, and saliva for DHT responses (moderate to large effect size). Baseline TT and FT correlated with oxygen uptake and work capacity in both genders (P < .05).
CONCLUSION
DHT showed no acute performance correlation but was responsive to volume of training, particularly in females. This work informs on timelines and relationships of androgenic biomarkers in males and females across different training loads, adding to the complexity that should be considered in interpretation thereof. The authors speculate that testosterone may impact acute performance via behavioral mechanisms of motivation and attention; DHT, via training volume-induced androgenic promotion, may facilitate long-term adaptive changes, especially for females.
Topics: Athletes; Dihydrotestosterone; Exercise; Female; Humans; Male; Testosterone
PubMed: 33952710
DOI: 10.1123/ijspp.2020-0910 -
Reproductive Biology Mar 2022Oviduct ampulla plays an important role in steroid hormone-regulated sperm-oocyte binding in female animals. Although studies have shown that androgen receptor are...
Oviduct ampulla plays an important role in steroid hormone-regulated sperm-oocyte binding in female animals. Although studies have shown that androgen receptor are expressed in many species oviduct, the interaction among androgen receptor (AR), estradiol (E2) and progesterone (P4) in the sheep oviduct have rarely been reported. In this study, we evaluated the localization of two isoforms of dihydrotestosterone (DHT) sythetase enzymes 5α-reductase (5α-red1, 5α-red2) and AR in sheep oviduct ampulla by immunohistochemistry and immunofluorescence. Results showed that they were all distributed in oviduct epithelium layer. In epithelial cells, 5α-red1, 5α-red2 were expressed in cytoplast and nuclear, but AR were stained in nuclear. We also investigated their expression pattern in the sheep oviduct ampulla at different development stages of follicles (Large follicles stage; hemorrhagium, luteum and albicans of corpus stage) by molecular experiments. We found that 5α-red1, 5α-red2 and AR mRNA abundance and protein were expressed highest in corpus albicans stage and lowest in corpus hemorrhagium stage. In vitro, when sheep oviduct ampulla epithelial cells (SOAECs) were cultured and treated with different concentrations of E2/P4 (10-10 M), we found that E2 inhibited the expression of AR mRNA and protein, while P4 promoted this expression. In addition, when the SOAECs were treated with E2 (10 M) and/or its non-selective inhibitor ICI182780 (10 M) as well as with P4 (10 M) and/or its non-specific inhibitor RU486 (10 M), we found that E2 and P4 inhibited and promoted the expression of AR mRNA and proteins, respectively, via their nuclear receptor pathways. This study provides a basic insight for the further research of oviduct epithelium physiological function closely related to androgen.
Topics: Animals; Dihydrotestosterone; Estradiol; Female; Humans; Oviducts; Progesterone; Receptors, Androgen; Sheep
PubMed: 35114486
DOI: 10.1016/j.repbio.2021.100573 -
Cellular and Molecular Biology... Jan 2024Neuroinflammation induced by microglia following spinal cord injury (SCI) leads to secondary neurologic injury. Androgens including testosterone and dihydrotestosterone...
Neuroinflammation induced by microglia following spinal cord injury (SCI) leads to secondary neurologic injury. Androgens including testosterone and dihydrotestosterone (DHT) show as endogenous neuroprotective factors against multiple neurologic diseases, while their therapeutic role in SCI-induced neuroinflammation and underlying mechanism remains elusive. In the study, we aimed to investigate the role of DHT against microglia-induced neuroinflammation in SCI and evaluate its protective treatment. BV2 cells were activated by neuroinflammation via LPS in vitro. Adult male C57BL/6 mice were used to establish the SCI model. BV2 cells and SCI mice were administrated DHT. Microglia activation, pro-inflammatory factors, p38 and p65 phosphorylation, glial scar, fibrotic scar, histology, and locomotor function recovery were measured, respectively. We demonstrated that DHT administration attenuates neuroinflammation in microglia through inhibition of p38 and p65 pathways. Moreover, DHT reduces microglia and astrocyte accumulation, cord fibrosis and histologic damage. Besides, DHT ameliorates locomotor functional recovery after SCI. DHT is verified to play a neuroprotective role in SCI, which fights against neuroinflammation by inhibition of p38 and p65 pathways. Therefore, Mel is defined as a promising factor in protecting neural tissue after SCI.
Topics: Animals; Male; Mice; Dihydrotestosterone; Inflammation; Mice, Inbred C57BL; Microglia; Neuroinflammatory Diseases; NF-kappa B; Spinal Cord; Spinal Cord Injuries
PubMed: 38372091
DOI: 10.14715/cmb/2024.70.1.29 -
International Journal of Molecular... Jun 2022Skeletal muscle is a tissue that has recently been recognized for its ability to produce androgens under physiological conditions. The steroidogenesis process is known...
Skeletal muscle is a tissue that has recently been recognized for its ability to produce androgens under physiological conditions. The steroidogenesis process is known to be negatively influenced by reactive oxygen species (ROS) in reproductive Leydig and ovary cells, while their effect on muscle steroidogenesis is still an unexplored field. Muscle cells are continuously exposed to ROS, resulting from both their metabolic activity and the surrounding environment. Interestingly, the regulation of signaling pathways, induced by mild ROS levels, plays an important role in muscle fiber adaptation to exercise, in a process that also elicits a significant modulation in the hormonal response. The aim of the present study was to investigate whether ROS could influence steroidogenesis in skeletal muscle cells by evaluating the release of testosterone (T) and dihydrotestosterone (DHT), as well as the evaluation of the relative expression of the key steroidogenic enzymes 5α-reductase, 3β-hydroxysteroid dehydrogenase (HSD), 17β-HSD, and aromatase. C2C12 mouse myotubes were exposed to a non-cytotoxic concentration of hydrogen peroxide (HO), a condition intended to reproduce, in vitro, one of the main stimuli linked to the process of homeostasis and adaptation induced by exercise in skeletal muscle. Moreover, the influence of tadalafil (TAD), a phosphodiesterase 5 inhibitor (PDE5i) originally used to treat erectile dysfunction but often misused among athletes as a "performance-enhancing" drug, was evaluated in a single treatment or in combination with HO. Our data showed that a mild hydrogen peroxide exposure induced the release of DHT, but not T, and modulated the expression of the enzymes involved in steroidogenesis, while TAD treatment significantly reduced the HO-induced DHT release. This study adds a new piece of information about the adaptive skeletal muscle cell response to an oxidative environment, revealing that hydrogen peroxide plays an important role in activating muscle steroidogenesis.
Topics: Animals; Dihydrotestosterone; Female; Humans; Hydrogen Peroxide; Male; Mice; Muscle Fibers, Skeletal; Muscle, Skeletal; Reactive Oxygen Species; Testosterone
PubMed: 35743011
DOI: 10.3390/ijms23126566 -
Gynecologic Investigation 1971
Review
Topics: Animals; Chorionic Gonadotropin; Dihydrotestosterone; Dogs; Follicle Stimulating Hormone; Leydig Cells; Male; Prostate; Rats; Testis; Testosterone; Time Factors; Tritium
PubMed: 4949824
DOI: 10.1159/000301864 -
Imaging Androgen Receptors in Breast Cancer with F-Fluoro-5α-Dihydrotestosterone PET: A Pilot Study.Journal of Nuclear Medicine : Official... Jan 2022Most breast cancers express androgen receptors (ARs). This prospective imaging substudy explored imaging of ARs with F-fluoro-5α-dihydrotestosterone (F-FDHT) PET in...
Most breast cancers express androgen receptors (ARs). This prospective imaging substudy explored imaging of ARs with F-fluoro-5α-dihydrotestosterone (F-FDHT) PET in patients with metastatic breast cancer (MBC) receiving selective AR modulation (SARM) therapy (GTx-024). Eleven postmenopausal women with estrogen receptor-positive MBC underwent F-FDHT PET/CT at baseline and at 6 and 12 wk after starting SARM therapy. Abnormal tumor F-FDHT uptake was quantified using SUV AR status was determined from tumor biopsy specimens. F-FDHT SUV percentage change between scans was calculated. Best overall response was categorized as clinical benefit (nonprogressive disease) or progressive disease using RECIST 1.1. The median baseline F-FDHT SUV was 4.1 (range, 1.4-5.9) for AR-positive tumors versus 2.3 (range, 1.5-3.2) for AR-negative tumors ( = 0.22). Quantitative AR expression and baseline F-FDHT uptake were weakly correlated (Pearson ρ = 0.39, = 0.30). Seven participants with clinical benefit at 12 wk tended to have larger declines in F-FDHT uptake than did those with progressive disease both at 6 wk after starting GTx-024 (median, -26.8% [range, -42.9% to -14.1%], vs. -3.7% [range,-31% to +29%], respectively; = 0.11) and at 12 wk after starting GTx-024 (median, -35.7% [range, -69.5% to -7.7%], vs. -20.1% [range, -26.6% to +56.5%], respectively; = 0.17). These hypothesis-generating data suggest that F-FDHT PET/CT is worth further study as an imaging biomarker for evaluating the response of MBC to SARM therapy and reiterate the feasibility of including molecular imaging in multidisciplinary therapeutic trials.
Topics: Dihydrotestosterone
PubMed: 34049982
DOI: 10.2967/jnumed.121.262068 -
Journal of Fish Biology Dec 2023Sex steroids are known to modulate immune responses and as a result many of the immune parameters in seasonally breeding organisms show reproductive-phase dependent...
Sex steroids are known to modulate immune responses and as a result many of the immune parameters in seasonally breeding organisms show reproductive-phase dependent variation. Androgens, the male sex steroids, are largely reported to be immunosuppressive. Together with other pattern recognition receptors, the nucleotide-binding and oligomerization domain-like receptors (NLRs) serve as intracellular sentinels and are essential to defense mechanisms. Interestingly, to date the transcriptional modulation of NLRs by androgens has not been explored. In the present study, we investigated the reproductive-phase dependent expression of NLRs in the male spotted snakehead Channa punctata. Furthermore, the effect of dihydrotestosterone (DHT) on NLR expression was studied. The expression of NLRs was observed to be most pronounced during the spawning phase of the fish, which is marked by the highest testosterone level. In vivo as well as in vitro studies showed the diverse effect of DHT on NLR expression depending on the duration and mode of treatment, as well as the immune tissue studied.
Topics: Male; Animals; Dihydrotestosterone; Channa punctatus; Gene Expression; Phagocytosis; Androgens; Nucleotides
PubMed: 37641389
DOI: 10.1111/jfb.15546 -
European Journal of Medicinal Chemistry Mar 2023High expression of the androgen receptor (AR) and the disruption of its regulation are strongly responsible for the development of prostate cancer (PCa). Therapeutically...
High expression of the androgen receptor (AR) and the disruption of its regulation are strongly responsible for the development of prostate cancer (PCa). Therapeutically relevant non-steroidal or steroidal antiandrogens are able to block the AR effect by eliminating AR-mediated signalling. Herein we report the synthesis of novel steroidal pyrazoles derived from the natural sex hormone 5α-dihydrotestosterone (DHT). 2-Ethylidene or 2-(hetero)arylidene derivatives of DHT obtained by regioselective Claisen-Schmidt condensation with acetaldehyde or (hetero)aromatic aldehydes in alkaline ethanol were reacted with monosubstituted hydrazines to give A-ring-fused 1,5-disubstituted pyrazoles as main or exclusive products, depending on the reaction conditions applied. Spontaneous or 2,3-dichloro-5,6-dicyanobenzoquinone (DDQ)-induced oxidation of the primarily formed pyrazolines resulted in the desired products in moderate to good yields, while 17-oxidation also occurred by using the Jones reagent as a strong oxidant. Transcriptional activity of the AR in a reporter cell line was examined for all novel compounds, and several previously synthesized similar DHT-based pyrazoles with differently substituted heteroring were also included to obtain information about the structure-activity relationship. Two specific regioisomeric groups of derivatives significantly diminished the transcriptional activity of the AR in reporter cell line in 10 μM concentration, and displayed reasonable antiproliferative activity in AR-positive PCa cell lines. Lead compound (3d) was found to be a potent AR antagonist (IC = 1.18 μM), it generally suppressed AR signalling in time and dose dependent manner, moreover, it also led to a sharp decrease in wt-AR protein level probably caused by proteasomal degradation. We confirmed the antiproliferative activity of 3d in AR-positive PCa cell lines (with GI in low micromolar ranges), and its cellular, biochemical and in silico binding in AR ligand-binding domain. Moreover, compound 3d was shown to be potent even ex vivo in patient-derived tissues, which highlights the therapeutic potential of A-ring-fused pyrazoles.
Topics: Male; Humans; Dihydrotestosterone; Receptors, Androgen; Pyrazoles; Down-Regulation; Cell Line, Tumor; Prostatic Neoplasms; Steroids
PubMed: 36682291
DOI: 10.1016/j.ejmech.2023.115086