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1.
Artigo em Inglês | MEDLINE | ID: mdl-32109748

RESUMO

The standard approach to detect misuse with testosterone in sport is based on the determination and evaluation of the urinary steroid profile followed by the confirmation of atypical profiles using isotope ratio mass spectrometry. The detection capacity of these methods can be attenuated by confounding factors or testosterone preparations with endogenous isotopic fingerprints. An alternative detection method for misuse of an endogenous steroid in sports is the direct detection of the administered steroid ester present in most preparations. Thus unambiguous proof for doping misuse can be delivered. In this work, the sensitivity of gas chromatography coupled to a triple quadrupole with chemical ionization (GC-CI-MS/MS) is applied to detect trace levels of 10 testosterone and 2 nandrolone esters in plasma for in human doping analysis. The detection method was developed employing a liquid-liquid extraction and HPLC cleanup step before analysis on the GC-CI-MS/MS. The quantitative method was validated in a linear range of 100-2000 pg/ml and proved to be selective, reproducible and very sensitive with limits of detection as low as to 10 pg/ml. A clinical study with the administration of testosterone undecanoate in 3 volunteers was carried out and the compound was detectable up to 86 days after administration.


Assuntos
Doping nos Esportes , Cromatografia Gasosa-Espectrometria de Massas/métodos , Esteroides/sangue , Espectrometria de Massas em Tandem/métodos , Ésteres , Humanos , Modelos Lineares , Extração Líquido-Líquido , Masculino , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Esteroides/química , Esteroides/isolamento & purificação , Testosterona/análogos & derivados , Testosterona/sangue
2.
Int J Med Mushrooms ; 21(6): 523-536, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31679225

RESUMO

The Astraeus genus (Diplocystaceae) of ectomycorrhizal edible mushrooms is represented by nine species: A. asiaticus, A. hygrometricus, A. koreanus, A. morganii, A. odoratus, A. pteridis, A. sirindhorniae, A. smithii, and A. telleriae. Astraeus mushrooms, because of their characteristic delicacy and aroma, are marketed in several countries. Chemical examinations of these mushrooms have revealed their nutritional properties and bioactive constituents. Here, the proximate nutritional composition of A. hygrometricus and A. odoratus, and chemistry and biological activity of A. asiaticus, A. hygrometricus, A. odoratus, and A. pteridis were reviewed. Several mycochemicals, including polysaccharides, terpenoids, steroids, phenolics and heterocyclic compounds, have been characterized in their fruiting body. Various biological activities of these compounds are also discussed.


Assuntos
Agaricales/química , Basidiomycota/química , Valor Nutritivo , Fenóis/química , Polissacarídeos/química , Esteroides/química , Terpenos/química
3.
Molecules ; 24(20)2019 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-31614780

RESUMO

A small and focused library of steroidal non-fused and fused pyrimidines was prepared from pregnenolone acetate and diosgenin, respectively. The key step was the cycloaddition reaction of nitrogen-containing 1,3-binucleophiles with the steroidal α,ß-unsaturated ketone. Urea, thiourea and guanidine reacted in a similar manner and afforded the steroidal pyrimidines in good yields. The antiproliferative tests against human tumor cell lines gave GI50 values in the micromolar range and had no effect on healthy fibroblasts. Additional experiments indicated that the compounds did not act as P-glycoprotein substrates, thus avoiding the rise of drug resistance. The fused steroidal pyrimidinethione was selected as drug lead for further testing due to its strong antiproliferative activities within the low micromolar range.


Assuntos
Proliferação de Células/efeitos dos fármacos , Neoplasias/tratamento farmacológico , Pirimidinas/farmacologia , Esteroides/farmacologia , Acetatos/química , Linhagem Celular Tumoral , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Nitrogênio/química , Pregnenolona/química , Pirimidinas/síntese química , Pirimidinas/química , Esteroides/síntese química , Esteroides/química , Relação Estrutura-Atividade
4.
Phytochemistry ; 168: 112109, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31494344

RESUMO

Eight undescribed ergostane-type steroids, (22E,24R)-ergosta-7,22-dien-3ß,5α-diol- 6,5-olide, (22E,24R)-ergosta-7,9(11),22-trien-3ß,5ß,6ß-triol, (22E,24R)-6ß-methoxy ergosta-7,9(11),22-trien-3ß,5α,14ß-triol, (22E,24R)-9α,15α-dihydroxyergosta-4,6,8 (14),22-tetraen-3-one, (22E,24R)-ergosta-5,8,22-trien-3ß,11α-dihydroxyl-7-one, (22E,24R)-ergosta-4,7,22-trien-3ß,9α,14ß-trihydroxyl-6-one, (22E,24R)-ergosta-7,22- dien-3ß,9α,14ß-trihydroxyl-6-one, and (22E,24R)-6ß-methoxyergosta-7,22-dien-3ß, 5α,9α,14ß-tetraol, and twenty-one known analogues were isolated from the fruiting bodies of Ganoderma resinaceum Boud. Their chemical structures were determined on the basis of comprehensive spectroscopic analysis and X-ray crystal diffraction, as well as empirical pyridine-induced deshielding effects. Furthermore, selected compounds were evaluated for their inhibitory effects on macrophage activation using an inhibition of nitric oxide production assay. Finally, (22E,24R)-ergosta-5,8,22- trien-3ß,11α-dihydroxyl-7-one, (22E,24R)-ergosta-4,7,22-trien-3ß,9α,14ß-tri hydroxyl-6-one, (22E,24R)-6ß-methoxyergosta-7,22-dien-3ß,5α,9α,14ß-tetraol, (22E,24R)-ergosta-6,9,22-trien-3ß,5α,8α-triol,ergost-6,22-dien-3ß,5α,8α-triol, 5α,6α-epoxy-(22E,24R)-ergosta-8,22-diene-3ß,7α-diol, 5α,6α-epoxy-(22E,24R)- ergosta-8(14),22-diene-3ß,7α-diol, 5α,6α-epoxy-(22E,24R)-ergosta-8(14),22-diene-3ß, 7ß-diol, and 22E-7α-methoxy-5α,6α-epoxyergosta-8(14),22-dien-3ß-ol showed inhibitory effects on NO production with IC50 values ranging from 3.24 ±â€¯0.02 to 35.19 ±â€¯0.41 µM compared with L-NMMA (IC50 49.86 ±â€¯2.13 µM), indicating that they have potential anti-inflammatory activity.


Assuntos
Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/farmacologia , Carpóforos/química , Ganoderma/química , Esteroides/isolamento & purificação , Esteroides/farmacologia , Animais , Anti-Inflamatórios/química , Relação Dose-Resposta a Droga , Lipopolissacarídeos/antagonistas & inibidores , Lipopolissacarídeos/farmacologia , Camundongos , Conformação Molecular , Óxido Nítrico/antagonistas & inibidores , Óxido Nítrico/biossíntese , Células RAW 264.7 , Esteroides/química
5.
J Enzyme Inhib Med Chem ; 34(1): 1607-1614, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31474167

RESUMO

Steroids are classes of natural products widely distributed in nature, which have been demonstrated to exhibit broad biological functions, and have also attracted increasing interest from bioorganic and pharmaceutical researches. In order to develop novel chemical entities as potential cytotoxic agents, a series of steroidal isatin conjugations derived from epiandrosterone and androsterone were efficiently prepared and characterized, and all these obtained compounds were screened for their potential cytotoxic activities. The preliminary bioassay indicated that most of the newly synthesized compounds exhibited good cytotoxic activities against human gastric cancer (SGC-7901), melanoma (A875), and hepatocellular liver carcinoma (HepG2) cell lines compared with 5-fluorouracil (5-FU), which might be considered as promising scaffold for further development of potential anticancer agents.


Assuntos
Androsterona/química , Antineoplásicos Fitogênicos/farmacologia , Produtos Biológicos/farmacologia , Isatina/farmacologia , Esteroides/farmacologia , Androsterona/análogos & derivados , Antineoplásicos Fitogênicos/síntese química , Antineoplásicos Fitogênicos/química , Produtos Biológicos/síntese química , Produtos Biológicos/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Isatina/síntese química , Isatina/química , Estrutura Molecular , Esteroides/síntese química , Esteroides/química , Relação Estrutura-Atividade
6.
Diagn Microbiol Infect Dis ; 95(3): 114863, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31471074

RESUMO

It is known that synergy between Candida albicans and Staphylococcus aureus results in enhanced biofilm formation and increased resistance to antimicrobials. Ceragenins (CSAs) are derivatives of cholic acid designed to mimic the antimicrobial activities of endogenous antimicrobial peptides. In this study, various CSAs were tested on C. albicans and methicillin-susceptible S. aureus or methicillin-resistant S. aureus mono or multispecies biofilms at 2 different concentrations (16 and 64 µg/mL) and compared with conventional antimicrobials. CSA-8 was active agent both with mono and multispecies biofilms (P < 0.05). Among antifungals, amphotericin B and, among antibacterials, ciprofloxacin and gentamicin were active agents against all studied microorganisms. This study suggests that CSAs, especially CSA-8, have useful antibiofilm effects against monomicrobial or fungal-bacterial multispecies biofilms.


Assuntos
Anti-Infecciosos/farmacologia , Biofilmes/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Candida albicans/crescimento & desenvolvimento , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento , Esteroides/farmacologia , Biofilmes/crescimento & desenvolvimento , Técnicas de Cocultura , Contagem de Colônia Microbiana , Humanos , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Estrutura Molecular , Esteroides/química
7.
J Pharm Biomed Anal ; 175: 112756, 2019 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-31387028

RESUMO

In the context of hormonal contraception and hormone replacement therapy (HRT), many women are exposed to exogenous hormones. Current use of hormonal contraception with combined ethinyl estradiol and different progestins bestows a breast cancer relative risk (RR) of 1.2- while combined HRT has a RR of 2. Although these exposures present an important public health issue, little is known about the effects of individual progestins on the breast and other tissues. Increasing availability of large scale biobanks, high throughput analyses and data management tools enable ever expanding, sophisticated population studies. In order to address the impact of distinct progestins on various health indicators, it is desirable to accurately quantify progestins in clinical samples. Here we have developed and validated a high resolution liquid chromatography mass spectrometry (LC-MS) targeted method for the simultaneous quantification of 11 synthetic progestins widely used in oral contraceptives, gestodene, levonorgestrel, etonogestrel, chlormadinone acetate, cyproterone acetate, drospirenone, desacetyl norgestimate, medroxyprogesterone acetate, norethindrone, dienogest, nomegestrol acetate, and 4 endogenous steroid hormones, progesterone, testosterone, androstenedione, and cortisol in blood samples. This highly specific quantitative analysis with high resolution Orbitrap technology detects and quantifies 15 compounds using their internal standard counterparts in a single 12 min LC-MS run. Sensitivity is attained by the use of the instrument in targeted selected ion monitoring mode. Lower limit of quantitation ranges from 2.4 pg/ml for drospirenone to 78.1 pg/ml for chlormadinone acetate. The method provides comprehensive progestin panel measurements with as little as 50 µl of murine or human plasma.


Assuntos
Anticoncepcionais/química , Progestinas/química , Esteroides/química , Animais , Cromatografia Líquida/métodos , Feminino , Humanos , Camundongos , Camundongos Endogâmicos NOD , Espectrometria de Massas em Tandem/métodos
8.
Fitoterapia ; 137: 104281, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31381957

RESUMO

Veratrum californicum is a rich source of steroidal alkaloids, many of which have proven to be antagonists of the Hedgehog (Hh) signaling pathway that becomes aberrant in over twenty types of cancer. These alkaloids first became known in the 1950's due to their teratogenic properties, which resulted in newborn and fetal lambs developing cyclopia as a result of pregnant ewes consuming Veratrum californicum. It was discovered that the alkaloids in V. californicum were concentrated in the root and rhizome of the plant with much lower amounts of the most active alkaloid, cyclopamine, present in the aerial plant, especially in the late growth season. Inspired by the limitations in analytical instrumentation and methods available to researchers at the time of the original investigation, we have used state-of-the-art instrumentation and modern analytical methods to quantitate four steroidal alkaloids based on study parameters including plant part, harvest location, and growth stage. The results of the current inquiry detail differences in alkaloid composition based on the study parameters, provide a detailed assessment for alkaloids that have been characterized previously (cyclopamine, veratramine, muldamine and isorubijervine), and identify at least six alkaloids that have not been previously characterized. This study provides insight into optimal harvest time, plant growth stage, harvest location, and plant part required to isolate, yet to be characterized, alkaloids of interest for exploration as Hh pathway antagonists with desirable medicinal properties.


Assuntos
Alcaloides/química , Esteroides/química , Veratrum/química , Alcaloides/isolamento & purificação , Proteínas Hedgehog/antagonistas & inibidores , Idaho , Estrutura Molecular , Compostos Fitoquímicos/química , Compostos Fitoquímicos/isolamento & purificação , Componentes Aéreos da Planta/química , Rizoma/química , Estações do Ano , Esteroides/isolamento & purificação , Alcaloides de Veratrum
9.
Carbohydr Res ; 484: 107776, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31421353

RESUMO

Inhibiting effects of sulfated steroids from marine sponges of Halichondriidae family: halistanol sulfate, topsentiasterol sulfate D and chlorotopsentiasterol sulfate D were investigated on three different types of enzymes degrading polysaccharides of brown algae: endo-1,3-ß-d-glucanase GFA, fucoidan hydrolase FFA2 and bifunctional alginate lyase ALFA3 from marine bacterium Formosa algae KMM 3553T, inhabiting thalli of brown alga Fucus evanescens. This is the first research, devoted to influence of a marine natural compound on three functionally related enzymes that make up the complex of enzymes, necessary to degrade unique carbohydrate components of brown algae. Alginic acid, 1,3-ß-D-glucan (laminaran) and fucoidan jointly constitute practically all carbohydrate biomass of brown algae, so enzymes, able to degrade such polysaccharides, are crucial for digesting brown algae biomass as well as for organisms surviving and proliferating on brown algae thalli. Halistanol sulfate irreversibly inhibited native endo-1,3-ß-D-glucanases of marine mollusks, but reversibly competitively inhibited recombinant endo-1,3-ß-d-glucanase GFA. This fact indicates that there are significant structural differences between the enzymes of practically the same specificity. For alginate lyase and fucoidan hydrolase halistanol sulfate was irreversible inhibitor. Topsentiasterol sulfate D was less active inhibitor whereas chlorotopsentiasterol sulfate D was the strongest inhibitor of enzymes under the study. Chlorotopsentiasterol sulfate D caused 98% irreversible inhibition of GFA. Chlorotopsentiasterol sulfate D also caused reversible and 100% inhibition of ALFA3, which is unusual for reversible inhibitors. Inhibition of FFA2 was complete and irreversible in all cases.


Assuntos
Proteínas de Bactérias/antagonistas & inibidores , Flavobacteriaceae/enzimologia , Poríferos/química , Esteroides/farmacologia , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Proteínas de Bactérias/química , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Flavobacteriaceae/efeitos dos fármacos , Fucus/microbiologia , Hidrolases/antagonistas & inibidores , Simulação de Acoplamento Molecular , Estrutura Molecular , Polissacarídeo-Liase/antagonistas & inibidores , Polissacarídeos/química , Esteroides/química , Sulfatos/química
10.
Eur J Med Chem ; 181: 111520, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31404863

RESUMO

A series of novel triazole nucleobase analogues containing steroidal/coumarin/quinoline moieties have been synthesized based on copper-catalyzed azide-alkyne cycloaddition (CuAAC). The anti-cancer activity of the new triazole nucleobase analogues was studied in gastric cancer cell lines (MGC-803, SGC-7901) and normal gastric epithelial cells (GES-1) in vitro. Some of the synthesized compounds could significantly inhibit the proliferation of these tested cancer cells. Among the tested compounds, compound 20c demonstrated good anti-proliferation activity against MGC-803 cells (IC50 = 1.48 µM) and SGC-7901 (IC50 = 2.28 µM) cells as well as the best selectivity between the cancer and normal cells. Further mechanistic studies indicated that compound 20c could down-regulate the expression of TGF ß1 both in the tested gastric cancer cell lines and inhibit the cell migration and invasion. The results of the study indicate that compound 20c could be used as a promising skeleton for anti-gastric cancer agents with improved efficacy and less side effects.


Assuntos
Antineoplásicos/farmacologia , Cumarínicos/farmacologia , Quinolinas/farmacologia , Esteroides/farmacologia , Neoplasias Gástricas/tratamento farmacológico , Triazóis/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/química , Proliferação de Células/efeitos dos fármacos , Cumarínicos/química , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Estrutura Molecular , Quinolinas/química , Esteroides/química , Neoplasias Gástricas/patologia , Relação Estrutura-Atividade , Triazóis/síntese química , Triazóis/química , Células Tumorais Cultivadas
11.
Eur J Med Chem ; 179: 694-706, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31284080

RESUMO

Unique derivatives of androstene and estrane series containing N-sulfonylimidate pendants were prepared from 17α-ethynyl steroids via Cu-catalyzed azide-alkyne cycloaddition to tosyl azide in the presence of alcohols. The synthesized compounds were screened for cytotoxicity against human breast cancer cell lines and ERα agonist activity. The hit compound 3,17ß-dimethoxy-17α-[iso-propyl-2'-N-tosylacetimidate]estra-1,3,5(10)-triene (4n) had no ERα-mediated hormonal activity and was found to exhibit potent cytotoxic effect in an ERα-positive breast cancer cell line. N-Sulfonylimidate 4n displayed high antiproliferative potency against triple-negative MDA-MB-231 breast cancer cells, while it was non-toxic towards normal mammary epithelial cells. Compound 4n was found to alter activity of various signaling pathways (NF-κB, Slug, cyclin D1, ERK) supporting the growth and invasiveness of tumor cells.


Assuntos
Antineoplásicos/farmacologia , Imidoésteres/farmacologia , Esteroides/farmacologia , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Antineoplásicos/síntese química , Antineoplásicos/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Imidoésteres/síntese química , Imidoésteres/química , Células MCF-7 , Estrutura Molecular , Esteroides/síntese química , Esteroides/química , Relação Estrutura-Atividade , Neoplasias de Mama Triplo Negativas/patologia
12.
Nat Commun ; 10(1): 3378, 2019 07 29.
Artigo em Inglês | MEDLINE | ID: mdl-31358750

RESUMO

Steroidal C19-hydroxylation is pivotal to the synthesis of naturally occurring bioactive C19-OH steroids and 19-norsteroidal pharmaceuticals. However, realizing this transformation is proved to be challenging through either chemical or biological synthesis. Herein, we report a highly efficient method to synthesize 19-OH-cortexolone in 80% efficiency at the multi-gram scale. The obtained C19-OH-cortexolone can be readily transformed to various synthetically useful intermediates including the industrially valuable 19-OH-androstenedione, which can serve as a basis for synthesis of C19-functionalized steroids as well as 19-nor steroidal drugs. Using this biocatalytic C19-hydroxylation method, the unified synthesis of six C19-hydroxylated pregnanes is achieved in just 4 to 9 steps. In addition, the structure of sclerosteroid B is revised on the basis of our synthesis.


Assuntos
Androstenodiona/química , Cortodoxona/química , Pregnanos/química , Esteroides/química , Androstenodiona/metabolismo , Biocatálise , Cortodoxona/metabolismo , Hidroxilação , Modelos Químicos , Estrutura Molecular , Pregnanos/metabolismo , Esteroides/síntese química , Esteroides/metabolismo
13.
Talanta ; 204: 415-423, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31357314

RESUMO

A cyclic-organophosphate, specifically 2-chloro-5,5-dimethyl-1,3,2-dioxaphosphorinane-2-oxide, was used to derivatise the hydroxyl group at the C3 position of selected steroid hormones to analyse the derivatives using UPLC-MS/MS (ultra-performance liquid chromatography-tandem mass spectrometry). Reactions were performed in an anhydrous pyridine environment in the presence of AlCl3 at 50 °C. The developed reaction is suitable for analytical chemistry applications and was validated by analysis of selected contraceptive drugs. The sensitivity of the method depends on hormone tested and the limit of detection ranges from 130 pg/mL for ß-estradiol to 240 pg/mL for estriol. The estimated efficiency of derivatisation reactions varies in the range from 77.5 to 95.7%, and depends upon the hormone undergoing derivatisation. The method's recovery rate for the lowest concentration tested (800 pg/mL) is 88.1-96.3%. The method exhibits linearity in the 390 pg/mL to 2.5 µg/mL range, with R2 = 0.997. The developed steroid hormone derivatisation reaction was validated experimentally using UHPLC-QTOF-MS (ultra-high performance liquid chromatography quadrupole time of flight mass spectrometry) and NMR (nuclear magnetic resonance) spectroscopy. These studies show that the developed derivatisation reaction provides a precise and repeatable determination of selected steroid hormones in contraceptive drugs. At n = 10, CV (Coefficient of Variation) did not exceed 7%, which is a very good result compared with other analytical methods.


Assuntos
Congêneres do Estradiol/análise , Indicadores e Reagentes/química , Organofosfatos/química , Esteroides/análise , Cromatografia Líquida de Alta Pressão/métodos , Congêneres do Estradiol/química , Limite de Detecção , Fosforilação , Esteroides/química , Espectrometria de Massas em Tandem/métodos
14.
Mar Drugs ; 17(8)2019 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-31357591

RESUMO

Seven new unusual polysulfated steroids-topsentiasterol sulfate G (1), topsentiasterol sulfate I (2), topsentiasterol sulfate H (3), bromotopsentiasterol sulfate D (4), dichlorotopsentiasterol sulfate D (8), bromochlorotopsentiasterol sulfate D (9), and 4ß-hydroxyhalistanol sulfate C (10), as well as three previously described-topsentiasterol sulfate D (7), chlorotopsentiasterol sulfate D (5) and iodotopsentiasterol sulfate D (6) have been isolated from the marine sponge Halichondria vansoesti. Structures of these compounds were determined by detailed analysis of 1D- and 2D-NMR and HRESIMS data, as well as chemical transformations. The effects of the compounds on human prostate cancer cells PC-3 and 22Rv1 were investigated.


Assuntos
Glucose/metabolismo , Poríferos/química , Antígeno Prostático Específico/metabolismo , Esteroides/química , Esteroides/farmacologia , Sulfatos/química , Sulfatos/farmacologia , Animais , Humanos , Espectroscopia de Ressonância Magnética/métodos , Células PC-3 , Esteróis/química , Esteróis/farmacologia , Vietnã
15.
BMC Biotechnol ; 19(1): 39, 2019 06 25.
Artigo em Inglês | MEDLINE | ID: mdl-31238923

RESUMO

BACKGROUND: Aerobic side chain degradation of phytosterols by actinobacteria is the basis for the industrial production of androstane steroids which are the starting materials for the synthesis of steroid hormones. A native strain of Mycobacterium sp. VKM Ac-1817D effectively produces 9α-hydroxyandrost-4-ene-3,17-dione (9-OH-AD) from phytosterol, but also is capable of slow steroid core degradation. However, the set of the genes with products that are involved in phytosterol oxidation, their organisation and regulation remain poorly understood. RESULTS: High-throughput sequencing of the global transcriptomes of the Mycobacterium sp. VKM Ac-1817D cultures grown with or without phytosterol was carried out. In the presence of phytosterol, the expression of 260 genes including those related to steroid catabolism pathways significantly increased. Two of the five genes encoding the oxygenase unit of 3-ketosteroid-9α-hydroxylase (kshA) were highly up-regulated in response to phytosterol (55- and 25-fold, respectively) as well as one of the two genes encoding its reductase subunit (kshB) (40-fold). Only one of the five putative genes encoding 3-ketosteroid-∆1-dehydrogenase (KstD_1) was up-regulated in the presence of phytosterol (61-fold), but several substitutions in the conservative positions of its product were revealed. Among the genes over-expressed in the presence of phytosterol, several dozen genes did not possess binding sites for the known regulatory factors of steroid catabolism. In the promoter regions of these genes, a regularly occurring palindromic motif was revealed. The orthologue of TetR-family transcription regulator gene Rv0767c of M. tuberculosis was identified in Mycobacterium sp. VKM Ac-1817D as G155_05115. CONCLUSIONS: High expression levels of the genes related to the sterol side chain degradation and steroid 9α-hydroxylation in combination with possible defects in KstD_1 may contribute to effective 9α-hydroxyandrost-4-ene-3,17-dione accumulation from phytosterol provided by this biotechnologically relevant strain. The TetR-family transcription regulator gene G155_05115 presumably associated with the regulation of steroid catabolism. The results are of significance for the improvement of biocatalytic features of the microbial strains for the steroid industry.


Assuntos
Androstenodiona/metabolismo , Proteínas de Bactérias/genética , Perfilação da Expressão Gênica/métodos , Mycobacterium/genética , Fitosteróis/farmacologia , Transcriptoma/efeitos dos fármacos , Androstenodiona/química , Proteínas de Bactérias/metabolismo , Sequência de Bases , Genoma Bacteriano/genética , Redes e Vias Metabólicas/genética , Modelos Químicos , Estrutura Molecular , Mycobacterium/metabolismo , Oxigenases/genética , Oxigenases/metabolismo , Homologia de Sequência do Ácido Nucleico , Esteroides/química , Esteroides/metabolismo , Transcriptoma/genética
16.
Molecules ; 24(13)2019 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-31247920

RESUMO

Sulfoconjugates of sterols play important roles as neurosteroids, neurotransmitters, and ion channel ligands in health and disease. In most cases, sterol conjugate analysis is performed with liquid chromatography-mass spectrometry. This is a valuable tool for routine analytics with the advantage of direct sterol sulfates analysis without previous cleavage and/or derivatization. The complementary technique gas chromatography-mass spectrometry (GC-MS) is a preeminent discovery tool in the field of sterolomics, but the analysis of sterol sulfates is hampered by mandatory deconjugation and derivatization. Despite the difficulties in sample workup, GC-MS is an indispensable tool for untargeted analysis and steroid profiling. There are no general sample preparation protocols for sterol sulfate analysis using GC-MS. In this study we present a reinvestigation and evaluation of different deconjugation and derivatization procedures with a set of representative sterol sulfates. The advantages and disadvantages of trimethylsilyl (TMS), methyloxime-trimethylsilyl (MO-TMS), and trifluoroacetyl (TFA) derivatives were examined. Different published procedures of sterol sulfate deconjugation, including enzymatic and chemical cleavage, were reinvestigated and examined for diverse sterol sulfates. Finally, we present a new protocol for the chemical cleavage of sterol sulfates, allowing for simultaneous deconjugation and derivatization, simplifying GC-MS based sterol sulfate analysis.


Assuntos
Cromatografia Gasosa-Espectrometria de Massas , Esteroides/química , Esteróis/química , Sulfatos/química , Humanos , Estrutura Molecular , Solventes , Esteroides/análise , Esteróis/análise , Sulfatos/análise
17.
Org Biomol Chem ; 17(24): 5925-5928, 2019 06 18.
Artigo em Inglês | MEDLINE | ID: mdl-31165123

RESUMO

A novel ergostane, sarocladione (1), was isolated from the deep-sea-derived fungus Sarocladium kiliense, along with 20 known compounds. The structure of 1 was determined mainly by a detailed analysis of its experimental and calculated NMR spectroscopic data. It is worth noting that 1 was the first steroid bearing a 5,10:8,9-diseco moiety. All 21 compounds were tested for in vitro antitumor activities against five cancer cell lines. ß-Sitostenone (7) and 4,6-dihydroxyeudesmane (20) showed significant effects on HeLa-S3 cells with the IC50 values of 9.2 µM and 9.3 µM, respectively.


Assuntos
Acremonium/química , Antineoplásicos/farmacologia , Esteroides/farmacologia , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Células HeLa , Humanos , Conformação Molecular , Esteroides/química , Esteroides/isolamento & purificação , Relação Estrutura-Atividade
18.
Phytochemistry ; 164: 172-183, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31158602

RESUMO

Screening assays showed that total glycoside-rich fraction (TG) of rhizomes of Polygonatum sibiricum unveiled remarkable anti-proliferative activities against three cancer cell lines (A549, HepG2, and Caco2). Activity-guided isolation of TG afforded seven undescribed steroidal glycosides (polygonosides 1-7), along with 24 known glycosides. Their structures were established by 1D and 2D NMR spectroscopic analyses, high-resolution mass spectrometry, and chemical evidence. The isolated steroidal glycosides were tested for their antiproliferative activities against A549, HepG2, and Caco2 cells. Compounds 8, 10, 11, and 16 possessed stronger anticancer activities against A549 cells than the positive control Bay (25.8 µM), with IC50 values ranging from 5.8 to 24.2 µM. Compound 10 reduced the expression of Blc-2 and pro-caspase3 and increased the production of Bax as determined by western blotting. Molecular docking experiment suggested that 10 bound stably to the BH3-binding groove of the Bcl-2 protein by hydrogen bond interactions. These compounds could be candidates for anticancer agents with cytotoxic activity.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Glicosídeos/farmacologia , Polygonatum/química , Rizoma/química , Esteroides/farmacologia , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/isolamento & purificação , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Glicosídeos/química , Glicosídeos/isolamento & purificação , Humanos , Conformação Molecular , Simulação de Acoplamento Molecular , Esteroides/química , Esteroides/isolamento & purificação , Relação Estrutura-Atividade
19.
Pak J Pharm Sci ; 32(2): 721-741, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31081788

RESUMO

Present review discuss the reported work on structures, origins and the potent biologically active natural products isolated from Genus Buddleja, which is known for having many important pharmacologically active substances. The Genus Buddleja have more than 100 species, many of them are distributed in Mediterranean and Asian regions. A very small number of common species of the Genus in majority of fruiting plants have been investigated for their biological potential. So for, isolation of about 153 or more new/novel chemical substances have been reported. Purposes of the review is to discuss the structurally established and pharmacologically significant natural substances from wide variety of different species of this genus. Traditionally, species of the genus are reported to be used for healing, treatment of liver diseases, bronchial complaints, preventing several other diseases by exhibiting diuretic properties, sedative functions, analgesic potential, antirheumatic actions, antimicrobial activities, anti hyperglycemic functions and antioxidant properties. In this review we will describe recently established medicinal chemistry aspects and complete list of phytoconstituents as well as their sources and reference.


Assuntos
Anti-Infecciosos/farmacologia , Antioxidantes/farmacologia , Buddleja/química , Parassimpatolíticos/farmacologia , Plantas Medicinais/química , Analgésicos/farmacologia , Animais , Anti-Infecciosos/química , Antioxidantes/química , Buddleja/metabolismo , Diuréticos/farmacologia , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Flavonoides/química , Flavonoides/isolamento & purificação , Humanos , Hipnóticos e Sedativos/farmacologia , Parassimpatolíticos/química , Plantas Medicinais/metabolismo , Esteroides/química , Esteroides/isolamento & purificação , Terpenos/química , Terpenos/isolamento & purificação
20.
Nutrients ; 11(5)2019 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-31117231

RESUMO

Non-alcoholic steatohepatitis (NASH) is a progressive, chronic, liver disease whose prevalence is growing worldwide. Despite several agents being under development for treating NASH, there are no drugs currently approved. The Farnesoid-x-receptor (FXR) and the G-protein coupled bile acid receptor 1 (GPBAR1), two bile acid activated receptors, have been investigated for their potential in treating NASH. Here we report that BAR502, a steroidal dual ligand for FXR/GPBAR1, attenuates development of clinical and liver histopathology features of NASH in mice fed a high fat diet (HFD) and fructose (F). By RNAseq analysis of liver transcriptome we found that BAR502 restores FXR signaling in the liver of mice feed HFD-F, and negatively regulates a cluster of genes including Srebf1 (Srepb1c) and its target genes-fatty acid synthase (Fasn) and Cell death-inducing DFF45-like effector (CIDE) genes, Cidea and Cidec-involved in lipid droplets formation and triglycerides storage in hepatocytes. Additionally, BAR502 increased the intestinal expression of Fgf15 and Glp1 and energy expenditure by white adipose tissues. Finally, exposure to BAR502 reshaped the intestinal microbiota by increasing the amount of Bacteroidaceae. In conclusion, we have shown that dual FXR/GPBAR1 agonism might have utility in treatment of NASH.


Assuntos
Dieta Hiperlipídica/efeitos adversos , Perfilação da Expressão Gênica , Hepatopatia Gordurosa não Alcoólica/induzido quimicamente , Receptores Citoplasmáticos e Nucleares/metabolismo , Receptores Acoplados a Proteínas-G/metabolismo , Esteroides/química , Animais , Ácidos e Sais Biliares/metabolismo , Fezes , Microbioma Gastrointestinal , Regulação da Expressão Gênica/efeitos dos fármacos , Ligantes , Metabolismo dos Lipídeos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Hepatopatia Gordurosa não Alcoólica/metabolismo , Receptores Citoplasmáticos e Nucleares/genética , Receptores Acoplados a Proteínas-G/genética , Esteroides/metabolismo
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