Brassinosteroids (BRs) are steroid hormones regulating various aspects of plant metabolism, including growth, development and stress responses. However, little is known about the mechanism of their impact on antioxidant systems and phospholipid turnover. Using tobacco plants overexpressing H+/Ca2+vacuolar Arabidopsis antiporter CAX1, we showed the role of Ca2+ ion balance in the reactive oxygen species production and rapid phosphatidic acid accumulation induced by exogenous BR. Combination of our experimental results with public transcriptomic and proteomic data revealed a particular role of Ca2+-dependent phospholipid metabolizing enzymes in BR signaling. Here we provide novel insights into the role of calcium balance and lipid-derived second messengers in plant responses to exogenous BRs and propose a complex model integrating BR-mediated metabolic changes with phospholipid turnover.
- Klíčová slova
- Antioxidant enzymes, Brassinosteroids, Calcium, Nicotiana tabacum, Phosphatidic acid,
- MeSH
- antioxidancia * metabolismus MeSH
- Arabidopsis metabolismus účinky léků MeSH
- brassinosteroidy * metabolismus farmakologie MeSH
- kyseliny fosfatidové * metabolismus MeSH
- reaktivní formy kyslíku metabolismus MeSH
- tabák * metabolismus MeSH
- vápník * metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antioxidancia * MeSH
- brassinosteroidy * MeSH
- kyseliny fosfatidové * MeSH
- reaktivní formy kyslíku MeSH
- vápník * MeSH
The first review article on steroid dimers by Li and Dias in 1997, followed by the second review and a book on steroid dimers by Nahar and Sarker in 2007 and 2012, respectively, covered steroid dimers reported until the end of 2010. Since then, there have been considerable amounts of research carried out on steroid dimers, prompting the need for another comprehensive review on this topic. Therefore, this present review appraises the literature published during the period 2011-2019 on various aspects of steroid dimers, including isolation from natural sources, synthesis and applications. A structured and systematic literature search was performed, using the key words: steroid dimer, steroidal dimer, dimeric steroid, bis-steroid, bis-steroidal conjugates, molecular umbrella, cephalostatins, ritterazines and crellastatins. Several databases like Web of Knowledge, Science Direct, PubMed and Google Scholar were consulted. During the period covered in this review, well over 200 new synthetic steroidal dimers, ring A-ring A connection being the major group, have been reported, only one natural steroid dimer has been isolated, and potential applications of steroid dimers in the treatment of cancers and tumors, and microbial infections have been indicated.
- Klíčová slova
- Anticancer, Bis-steroid, Cephalostatins, Crellastatins, Molecular umbrella, Natural products, Ritterazines, Steroids dimer, Synthesis,
- MeSH
- antiinfekční látky farmakologie MeSH
- antitumorózní látky farmakologie MeSH
- dimerizace MeSH
- steroidy chemie farmakologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- antiinfekční látky MeSH
- antitumorózní látky MeSH
- steroidy MeSH
This paper describes the synthesis of a new A-homo lactam D-homo lactone androstane derivative from dehydroepiandrosterone. To evaluate the impact of the introduction of nitrogen in the parental scaffold on biological activity, a new androstane enamide-type lactam derivative was prepared and characterized. The new compound as well as starting compounds were screened for cytotoxic, anti-angiogenic and anti-inflammatory activities using several human cancer cell lines (MCF-7, MDA-MB-231, PC3, CEM, G-361, HeLa), endothelial (HUVEC) and non-tumour (MRC-5 and BJ) cell lines. Strong cytotoxic and anti-inflammatory activity with a broad therapeutical window was demonstrated by the A-homo lactam D-homo lactone androstane derivative. The induction of apoptosis in treated PC3 cultures was confirmed using apoptotic morphology screening and a fluorescent double-staining method. New A-homo lactam D-homo lactone androstane derivative induced apoptosis more than the tested reference compounds, Formestane and Doxorubicin. An in silico ADME analysis showed that the compounds possess drug-like properties.
- Klíčová slova
- ADME analysis in silico, Anti-angiogenic activity, Anti-inflammatory activity, Apoptotic morphology, Cytotoxic activity, Structural analysis,
- MeSH
- androstany chemie izolace a purifikace farmakologie MeSH
- antiflogistika chemie izolace a purifikace farmakologie MeSH
- antitumorózní látky fytogenní chemie izolace a purifikace farmakologie MeSH
- apoptóza účinky léků MeSH
- E-selektin antagonisté a inhibitory biosyntéza MeSH
- kultivované buňky MeSH
- laktony chemie izolace a purifikace farmakologie MeSH
- léky antitumorózní - screeningové testy MeSH
- lidé MeSH
- molekulární konformace MeSH
- optické zobrazování MeSH
- proliferace buněk účinky léků MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- androstany MeSH
- antiflogistika MeSH
- antitumorózní látky fytogenní MeSH
- E-selektin MeSH
- laktony MeSH
Heptakis(2,3,6-tri-O-methyl)-β-cyclodextrin (2) and cholesterol form a water-soluble complex 3. We performed several NMR studies, particularly 1H, 13C, 2D NOESY and DOSY, at various temperatures on 500 and 950 MHz instruments. We discovered that the complex 3 is unstable above 57 °C in heavy water, while it is kinetically stable enough to be studied by NMR in detail at 1 °C. We demonstrated the formation of a face-to-face 2:1 complex with a binding constant of approximately 2.2 × 106 M-2.
- Klíčová slova
- Cholesterol, Complex, Cyclodextrin, NMR, Structure,
- MeSH
- beta-cyklodextriny MeSH
- cholesterol chemie MeSH
- magnetická rezonanční spektroskopie MeSH
- molekulární modely MeSH
- molekulární struktura MeSH
- rozpustnost MeSH
- teplota MeSH
- voda chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- beta-cyklodextriny MeSH
- cholesterol MeSH
- heptakis(2,3,6-tri-O-methyl)-beta-cyclodextrin MeSH Prohlížeč
- voda MeSH
Two valuable forensic tools based on enzyme-linked immunoassays (ELISAs) for the analysis of 17α-methylated steroids were developed using haptens of stanazolol and its conjugates with biotin. Haptens containing terminal carboxylic group were conjugated to bovine serum albumin (BSA), rabbit serum albumin (RSA) or ovalbumin (OVA). Eight batches of antisera (RAbs) obtained by immunization of rabbits were tested in an indirect competitive ELISA system using immobilization of RSA conjugate (RSA/hapten) and competitor immobilization of the biotinylated conjugate (AB-ELISA) to avidin (avidin/hapten). The best results were achieved with the RAb 212 antibodies in RSA/ST-3 and avidin/ST-10 assembled variants. For the RSA/ST-3 system, an IC50 of 0.3 ng/mL and a detection limit of 0.02 ng/mL were measured. In case of avidin/ST-10 variant, IC50 was of 3.9 ng/mL and a detection limit of 0.57 ng/mL were obtained. The effect of solvent was tested as well as the stability of coated microtiter plates over four-month period. The cross-reactivity of the developed assays with other anabolic steroids was tested and high sensitivity towards 17α-methylated steroids was observed. RSA/ST-3 assay showed significant cross-reactivity with 17α-methyltestosterone (81.2%), oxymetholone (30.4%), methandienone (10.0%) and methyl dihydrotestosterone (7.7%). Similarly, in the avidin/ST-10 assay, 17α-methyltestosterone (34.5%), mestanolone (32.1%), oxymetholone (22.7%), methandienone (14.2%), 9-dehydromethyltestosterone (12.5%) and oxandrolone (1.2%) exhibited high cross-reactivity. The functionality of the developed systems was verified by the successful identification of a series of 17α-methylated anabolic steroids in a set of real samples including pharmaceutical preparations seized by the Police of the Czech Republic on the black market.
- Klíčová slova
- 17α-methylated anabolics, ELISA, Stanazolol,
- MeSH
- antisérum MeSH
- ELISA * MeSH
- kalibrace MeSH
- kongenery testosteronu analýza MeSH
- králíci MeSH
- metylace MeSH
- molekulární konformace MeSH
- sérový albumin chemie MeSH
- skot MeSH
- soudní lékařství * MeSH
- stanozolol chemie MeSH
- stereoizomerie MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antisérum MeSH
- kongenery testosteronu MeSH
- sérový albumin MeSH
- stanozolol MeSH
Anorexia nervosa (AN) is associated with various alterations including the dysfunction of the HPA axis and consequently the hypercortisolemia and deficit in sex hormones but the comprehensive evaluation of changes in circulating steroids during the hospitalization of AN patients is lacking. We investigated the effect of realimentation of women with AN during hospitalization on 45 circulating steroids, the relationships between BMI, its change during hospitalization and physical activity, on one side and initial levels and their changes for two adipokines, circulating steroids, anorexia-specific (hunger, appetite and satiety), and anorexia non-specific symptoms (anxiety, depression fatigue, sleep, and body pain) on the other side. We included 33 women with anorexia who were hospitalized for 38(35, 44) days (median with quartiles). The increase of BMI from the initial value 15.2 (13.2, 16.6) kg/m2 was 1.69 (1.37, 2.66) kg/m2. The patients with more severe anorexia showed higher activity in 7β-, and 16α-hydroxylation of androgen precursors, which declined during hospitalization. Otherwise, the 7α-hydroxylation activity is higher in AN patients with less severe malnutrition and the ratio of 5-androstene-3β,7α,17β-triol to 5-androstene-3β,7β,17β-triol increased during the realimentation. Our data allow to speculate that the intensive 7β-, and 16α- and possibly also the 7α-hydroxylation of C19 Δ5 steroids participate in the pathophysiology of anorexia by additional catabolism of substrates available for synthesis of active androgens and estrogens. However, the question remains whether the synthetic analogues of 7α/β- and 16α-hydroxy-steroids prevent the catabolism of the sex steroid precursors, or further activate the "energy wasting" mitochondrial thermogenic metabolism.
- Klíčová slova
- Anorexia nervosa, Anxiety, Depression, Refeeding, Specific symptoms and physical activity, Steroid metabolome,
- MeSH
- dospělí MeSH
- hospitalizace MeSH
- index tělesné hmotnosti MeSH
- lidé MeSH
- mentální anorexie metabolismus psychologie MeSH
- mladý dospělý MeSH
- steroidy krev metabolismus MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladý dospělý MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- steroidy MeSH
Amides of betulinic acid with cystamine were synthesized to investigate their antimicrobial and antitumor activity, and their influence on the cell cycle and cell apoptosis. The former target amide (6) displayed cytotoxicity in CEM cell line after 72 h of treatment (IC50 = 3.0 ± 0.7 μM; TI = 20), and induced apoptosis by caspase-3/7 activation in CEM cells. The latter target amide (9) displayed antimicrobial activity against Streptococcus mutans (MIC 3.125 μM; MBC 3.125 μM) and Bacillus cereus (MIC 25 μM; MBC 25 μM). The achieved results demonstrate enhancing of their biological activity over that of the parent compounds. However, two intermediate compounds (2 and 7) displayed either considerable cytotoxicity (2; 7.5 ± 0.8 μM; TI = 10, against G361) or antimicrobial activity (7; both against Actinomyces odontolycus and Clostridium perfrigens with MIC 12.5 µM and MBC 12.5 µM). The experimental data were compared with the in silico calculated physico-chemical and ADME parameters of the target compounds, including successful intermediates.
- Klíčová slova
- Antimicrobial activity, Apoptosis, Betulinic acid, Cystamine, Cytotoxicity,
- MeSH
- Actinomyces účinky léků MeSH
- amidy chemie farmakologie MeSH
- antibakteriální látky chemická syntéza chemie farmakologie MeSH
- antitumorózní látky chemická syntéza chemie farmakologie MeSH
- apoptóza účinky léků MeSH
- Bacillus cereus účinky léků MeSH
- Clostridium účinky léků MeSH
- cystamin chemie farmakologie MeSH
- kultivované buňky MeSH
- kyselina betulinová MeSH
- léky antitumorózní - screeningové testy MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- molekulární konformace MeSH
- pentacyklické triterpeny MeSH
- proliferace buněk účinky léků MeSH
- Streptococcus mutans účinky léků MeSH
- triterpeny chemie farmakologie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- amidy MeSH
- antibakteriální látky MeSH
- antitumorózní látky MeSH
- cystamin MeSH
- kyselina betulinová MeSH
- pentacyklické triterpeny MeSH
- triterpeny MeSH
Using Arabidopsis thaliana wild type (WT) plants and diacylglycerol kinase knockouts (single mutants - dgk3, dgk1, dgk6; double mutants - dgk3dgk7, dgk5dgk6, dgk1dgk2) we observed that the inhibitor of brassinosteroid (BR) biosynthesis, brassinazole (BRZ), drastically decreased germination of dgk mutants under salt stress, while BRZ co-administration with 24-epibrassinolide (EBL) partially improved germination rates. We also observed a statistically significant decrease in alternative and cytochrome respiratory pathways in response to BRZ treatment under salinity conditions. We showed that production of the lipid second messenger phosphatidic acid (PA) is impaired in dgk mutants in response to EBL treatment and inhibitor of diacylglycerol kinase (DGK) - R59022. This study demonstrates that dgk mutants possess lower germination rates, lower total respiration rates, an alternative respiratory pathway and PA content under optimal and high salinity conditions in response to EBL treatment comparing to WT plants.
- Klíčová slova
- Alternative oxidase, Arabidopsis thaliana, Brassinosteroids, Diacylglycerol kinase, Phosphatidic acid, Salinity,
- MeSH
- Arabidopsis chemie metabolismus MeSH
- brassinosteroidy farmakologie MeSH
- diacylglycerolkinasa antagonisté a inhibitory nedostatek metabolismus MeSH
- kyseliny fosfatidové chemie metabolismus MeSH
- salinita MeSH
- semena rostlinná účinky léků růst a vývoj metabolismus MeSH
- triazoly farmakologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- brassinazole MeSH Prohlížeč
- brassinosteroidy MeSH
- diacylglycerolkinasa MeSH
- kyseliny fosfatidové MeSH
- triazoly MeSH
Neurosteroids are endogenous steroidal compounds that can modulate neuronal receptors. N-Methyl-D-aspartate receptors (NMDARs) are glutamate-gated, calcium-permeable ion channels that are of particular interest, as they participate in synaptic transmission and are implicated in various processes, such as learning, memory, or long-term neuronal potentiation. Positive allosteric modulators that increase the activity of NMDARs may provide a therapeutic aid for patients suffering from neuropsychiatric disorders where NMDAR hypofunction is thought to be involved, such as intellectual disability, autism spectrum disorder, or schizophrenia. We recently described a new class of pregn-5-ene and androst-5-ene 3β-dicarboxylic acid hemiesters (2-24) as potent positive modulators of NMDARs. Considering the recommended guidelines for the early stage development of new, potent compounds, we conducted an in vitro safety assessment and plasma stability screening to evaluate their druglikeness. First, compounds were screened for their hepatotoxicity and mitochondrial toxicity in a HepG2 cell line. Second, toxicity in primary rat postnatal neurons was estimated. Next, the ability of compounds 2-24 to cross a Caco-2 monolayer was also studied. Finally, rat and human plasma stability screening revealed an unforeseen high stability of the C-3 hemiester moiety. In summary, by using potency/efficacy towards NMDARs data along with toxicity profile, Caco-2 permeability and plasma stability, compounds 14 and 15 were selected for further in vivo animal studies.
- Klíčová slova
- Caco-2 permeability, Hepatotoxicity, Mitotoxicity, NMDA receptor hypofunction, Neurosteroid, Plasma stability,
- MeSH
- androstenoly krev chemie farmakologie MeSH
- buňky Hep G2 MeSH
- cholesterol krev chemie farmakologie MeSH
- estery krev chemie farmakologie MeSH
- krysa rodu Rattus MeSH
- kyseliny dikarboxylové krev chemie farmakologie MeSH
- lidé MeSH
- mentální retardace farmakoterapie metabolismus MeSH
- mitochondrie účinky léků metabolismus MeSH
- molekulární struktura MeSH
- nádorové buňky kultivované MeSH
- neurony účinky léků metabolismus MeSH
- neuroprotektivní látky krev chemie farmakologie MeSH
- poruchy autistického spektra farmakoterapie metabolismus MeSH
- potkani Wistar MeSH
- pregnenolon analogy a deriváty krev farmakologie MeSH
- receptory N-methyl-D-aspartátu antagonisté a inhibitory metabolismus MeSH
- schizofrenie farmakoterapie metabolismus MeSH
- stabilita léku MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- androst-5-en-3-ol MeSH Prohlížeč
- androstenoly MeSH
- cholesterol MeSH
- estery MeSH
- kyseliny dikarboxylové MeSH
- neuroprotektivní látky MeSH
- pregn-5-en-3beta-ol MeSH Prohlížeč
- pregnenolon MeSH
- receptory N-methyl-D-aspartátu MeSH
