Pregnane Steroids Research Articles

Three New Steroids from the Roots of Marsdenia tenacissima.

Three undescribed pregnane steroids, 12β-O-4-hydroxybenzoyl tenacigenin D (1), 12β-O-4-hydroxybenzoyl tenacigenin A (2), and 11α-nicotinoyl-17β-marsdenin (3), along with two known analogues (4 and 5), were isolated from the roots of Marsdenia tenacissima. Their structures were elucidated using one- and two-dimensional NMR, high-resolution electron ionization-mass spectrometry, single-crystal X-ray diffraction data, and experimental and density-functional-theory-calculated electronic circular dichroism measurements. All isolated compounds were evaluated for their cytotoxic activities against human lung cancer cells (A549), ovarian carcinoma cells (SKOV-3), gastric cancer cells (MGC 803) and breast cancer cells (MCF-7). Notably, 3 exhibited significant cytotoxic activity against both A549 (median inhibitory concentration (IC50) = 16.79 μM) and SKOV-3 (IC50 = 12.30 μM) cells while exhibiting moderate cytotoxicity on MGC803 and MCF-7 cells.

The isolation of novel pregnane steroids from Aglaia pachyphylla Miq. and the cytotoxicity against breast cancer cell lines (MCF-7).

Steroid groups isolated from many plants are known to play a significant role in various biological systems. Therefore, this research aimed to analyze two novel pregnane steroids, pachylenone A (1) and pachylenone B (2), isolated from Aglaia pachyphylla Miq. The cytotoxicity of the steroids was evaluated against MCF-7 breast cancer cell lines with other known steroid compounds, namely 5α-dihydroprogesterone (3), GSD-8 (4), trans-5α-pregn-l7(20)-en-3,16-dion (5), 20β-hydroxy-5αH-pregnan-3-one (6), 3β-hydroxy-5α-pregnan-20-one (7), aglaiasterol B (8), and 2β,3β-dihydroxypregnan-16-one (9). Meanwhile, structural elucidation was achieved through different spectroscopic methods including one and two-dimensional NMR, as well as mass spectroscopy and quantum chemical calculations (TD-DFT and NMR DP4+ probability). The cytotoxic effects of steroid compounds (1-9) on MCF-7 lines were also examined. The results showed that compound 8 had the strongest activity with an IC50 value of 228 μM, followed by compound 6 (IC50 568,76 μM), and pachylenone A (1) (IC50 768.73 μM). As a recommendation for future research, other activities of these compounds should be evaluated.

Unusual triterpenoids and steroids from Cipadessa baccifera and their biological activities

Five undescribed triterpenoids and steroids (1–5), as well as ten known compounds, were purified from the branches and leaves of Cipadessa baccifera. Notably, 1 and 2 are rare cipadesin-type limonoids with an unusual 8,30-epoxide ring and 1,8-ether linkage, respectively. Compound 5 possessed pregnane steroid skeleton with an uncommon 5/6/6/6/5-fused ring system. Their structures were constructed by extensive spectroscopic analysis (NMR, IR, UV, and HRESIMS), and their absolute configurations were confirmed by ECD calculations and quantum chemical calculations. All the isolates were in vitro assayed for their antimicrobial potentials against 6 pathogenic microorganisms and antiproliferation activities against five human cancer cell lines. As a result, compounds 5, 12, 13, and 14 exhibited moderate antibacterial activities (MIC: 25–50 μg/mL). Moreover, 5 showed cytotoxicity against five cancer cell lines with IC50 values ranging from 8.0 to 19.9 μM.

Toxicity and chemical compounds of Piper aduncum fruit extract against storage pest Sitophilus oryzae and Callosobruchus maculatus

Insect pests of rice weevil of Sitophilus oryzae and cowpea beetles of Callosobruchus maculatus can reduce the quality and quantity of stored products. Various alternatives development to control insect pests more safely were necessary. Botanical insecticides can be used as an alternative control because of their several advantages. One of the potential plants as a source for botanical insecticides is Piper aduncum (Piperaceae). This study aimed to determine the chemical constituents of ethyl acetate extract of P. aduncum fruit (Ea-Pa) and to assess its toxicity against S. oryzae and C. maculatus female adults by contact residue. The most abundant chemical compound of Ea-Pa was dillapiol (61.54%). Dillapiol was one compound that might be responsible for the insecticidal properties, as well as ylangene, naphthalene, piperitone, hexadecanoic acid, β-eudesmene, α-cubebene, δ-cadinene, myristicin, cis-vaccenic acid, pregnane steroids, octacosane, pyridine, sitosterol, and caryophyllene. The toxicity assay results showed that the Ea-Pa was more toxic against female adults of C. maculatus than S. oryzae. The LC50 values for female adults of S. oryzae and C. maculatus were 4.05% and 0.17%, whereas LC95 values were 116.40% and 4.95%, respectively. Therefore, the Ea-Pa could be further improved as a natural insecticide to manage pest insects in storage.

Pregnane steroid glycosides with multidrug resistance reversal activity from the stems of Marsdenia tenacissima

Eighteen previously unreported pregnane glycosides, namely marsdenosides S1–S18, along with 15 known analogues, have been isolated from the stems of Marsdenia tenacissima. The structures of the undescribed compounds were elucidated by spectroscopic means, and their absolute configurations were established on the basis of time-dependent density functional theory (TD-DFT) based electronic circular dichroism (ECD) calculation, X-ray crystallography and acid hydrolysis. All the isolates were evaluated for their chemo-reversal ability against P-glycoprotein (P-gp)-mediated multidrug resistance (MDR) in MCF-7/ADR cell line, and nine ones displayed moderate MDR reversal activity with reversal folds in the range of 2.45–9.01. The most active 12-O-acetyl-20-O-benzoyl-(14,17,18-orthoacetate)-dihydrosarcostin-3-O-β-d-thevetopyranosyl-(1 → 4)-O-β-d-oleandropyranosyl-(1 → 4)-O-β-d-cymaropyranoside increased the sensitivity of MCF-7/ADR cell to adriamycin comparably to the reference drug verapamil (RF = 8.93).

Pregnane steroids and steroid glycosides with multidrug resistance reversal activity from Marsdenia tenacissima

Twenty compounds comprising four pregnane steroids (2-4 & 20) and 16 pregnane glycosides (1 & 5-19) have been obtained from the ethanol extract of the roots of a Dai ethnological herb, Marsdenia tenacissima. Their structures were characterized on the basis of comprehensive spectroscopic analyses with 17 ones (1-17) being reported for the first time, including the rare cases (2 & 3) of free C21 steroids with 17α-acetyl substitution, compounds 4-7 bearing an unusual 14α-OH, and the first examples with simultaneous 14α-OH/17α-acetyl substitution (7) and glycosylation at C-12 position (10 & 11). An empirical rule for the identification of C-17 configuration, in C21 steroids incorporating the marsdenin constitution structure, was also proposed. All the isolates, along with an array of previously reported analogues in our compound library, were screened for their chemo-reversal ability against P-glycoprotein (P-gp)-mediated multidrug resistance (MDR) in MCF-7/ADR cell line, and six compounds exhibited moderate MDR reversal activity with reversal folds ranging from 1.92 to 4.44.

Jasminanthoside – a new pregnane-steroid from Jasminanthes tuyetanhiae

A new pregnane steroid, jasminanthoside (1), together with three known compounds, telosmoside A7 (2), syringaresinol (3), and methyl 6–deoxy–3–O–methyl–β–D–allopyranosyl–(1→4)–β–D–oleandropyranoside (4) were isolated from the ethyl acetate extract of Jasminanthes tuyetanhiae roots collected in Vietnam. Their chemical structures were elucidated by NMR and MS spectroscopic data analysis along with the comparison of their data with the published ones in the literature. Although 4 was a known compound, its full NMR data were reported for the first time. All isolated compounds, evaluated for their α-glucosidase inhibition, showed activities stronger than the positive control acarbose. Among them, 1 was the best with the IC50 value of 7.41 ± 0.59 µM.

Tuberculosis alters immune-metabolic pathways resulting in perturbed IL-1 responses.

Tuberculosis (TB) remains a major public health problem and we lack a comprehensive understanding of how Mycobacterium tuberculosis (M. tb) infection impacts host immune responses. We compared the induced immune response to TB antigen, BCG and IL-1β stimulation between latently M. tb infected individuals (LTBI) and active TB patients. This revealed distinct responses between TB/LTBI at transcriptomic, proteomic and metabolomic levels. At baseline, we identified a novel immune-metabolic association between pregnane steroids, the PPARγ pathway and elevated plasma IL-1ra in TB. We observed dysregulated IL-1 responses after BCG stimulation in TB patients, with elevated IL-1ra responses being explained by upstream TNF differences. Additionally, distinct secretion of IL-1α/IL-1β in LTBI/TB after BCG stimulation was associated with downstream differences in granzyme mediated cleavage. Finally, IL-1β driven signalling was dramatically perturbed in TB disease but was completely restored after successful treatment. This study improves our knowledge of how immune responses are altered during TB disease, and may support the design of improved preventive and therapeutic tools, including host-directed strategies.

Isolation of a new steroid from phytochemical investigation on Caralluma lasiantha

A new steroid (acylated C21 pregnane steroid) was isolated from chloroform extract in phytochemical screening of Caralluma lasiantha. The isolated compound is found to be 3β-hydroxy-14β–(6′- carboxyphenyl)propionyloxypregn–5-en-20-one based on spectroscopic studies (IR, 1H NMR, 13C NMR, DEPT, COSY, HSQC, HMBC and ESI-MS). The isolated new steroid was tested against four bacterial strains and the activity was related to the structure of the molecule.

Steroid hormones of the octopus self-destruct system

Among all invertebrates, soft-bodied cephalopods have the largest central nervous systems and the greatest brain-to-body mass ratios, yet unlike other big-brained animals, cephalopods are unusually short lived.1-5 Primates and corvids survive for many decades, but shallow-water octopuses, such as the California two-spot octopus (Octopus bimaculoides), typically live for only 1 year.6,7 Lifespan and reproduction are controlled by the principal neuroendocrine center of the octopus: the optic glands, which are functional analogs to the vertebrate pituitary gland.8-10 After mating, females steadfastly brood their eggs, begin fasting, and undergo rapid physiological decline, featuring repeated self-injury and leading to death.11 Removal of the optic glands completely reverses this life history trajectory,10 but the signaling factors underlying this major life transition are unknown. Here, we characterize the major secretions and steroidogenic pathways of the female optic gland using mass spectrometry techniques. We find that at least three pathways are mobilized to increase synthesis of select sterol hormones after reproduction. One pathway generates pregnane steroids, known in other animals to support reproduction.12-16 Two other pathways produce 7-dehydrocholesterol and bile acid intermediates, neither of which were previously known to be involved in semelparity. Our results provide insight into invertebrate cholesterol pathways and confirm a remarkable unity of steroid hormone biology in life history processes across Bilateria.

Pregnane steroids from the twigs and leaves of Strophanthus divaricatus and their cytotoxic activities

Four new pregnanes (1–4), including one 21-norpregnane (4), together with two known pregnanes (5 and 6), were isolated from the twigs and leaves of Strophanthus divaricatus. Their structures were elucidated on the basis of spectroscopic data analysis, ECD data, and X-ray crystallographic study. The cytotoxic activities of these pregnanes against two human cancer cell lines were evaluated.

Mining Anti-Inflammation Molecules From Nippostrongylus brasiliensis-Derived Products Through the Metabolomics Approach.

Hookworm is one type of soil-transmitted helminth, which could exert an anti-inflammatory effect in human or animal host, which provides a beneficial possibility for the discovery of inflammatory-related disease interventions. The identification of hookworm-derived anti-inflammatory molecules is urgently needed for future translational research. The emergence of metabolomics has become a powerful approach to comprehensively characterize metabolic alterations in recent times. Herein, excretory and secretory products (ESPs) were collected from cultured adult worm, while small intestinal contents were obtained from Nippostrongylus brasiliensis (N. brasiliensis, Nb)-infected mice. Through ultra-high-performance liquid chromatography coupled with mass spectrometry (UHPLC-MS) platform, metabolomics analysis was used to explore the identification of anti-inflammatory molecules. Out of 45 differential metabolites that were discovered from ESPs, 10 of them showed potential anti-inflammatory properties, which could be subclassed into amino acids, furanocoumarins, linear diarylheptanoids, gamma butyrolactones, and alpha-keto acids. In terms of intestinal contents that were derived from N. brasiliensis-infected mice, 14 out of 301 differential metabolites were discovered to demonstrate anti-inflammatory effects, with possible subclassification into amino acids, benzylisoquinolines, quaternary ammonium salts, pyrimidines, pregnane steroids, purines, biphenyls, and glycerophosphocholines. Furthermore, nine of the differential metabolites appeared both in ESPs and infected intestinal contents, wherein four were proven to show anti-inflammation properties, namely, L-glutamine, glutamine (Gln), pyruvate, and alanine-Gln (Ala-Gln). In summary, we have provided a method for the identification and analysis of parasite-derived molecules with potential anti-inflammatory properties in the present study. This array of anti-inflammatory metabolites could provide clues for future evaluation and translational study of these anti-inflammatory molecules.

Pregnane Steroids from the Leaves of Melia Azedarach and Apoptotic Activity against T47D Cells.

Objective:Nature has provided us with many pharmaceutical resources so far. Breast cancer shows an increasing trend in the world for the last decade and becomes one of five leading causes of death. Among the plants, Melia azedarach L. has been used widely in traditional medicine for many ailments including breast cancer. Following our previous findings that the ethyl acetate fraction was the most active cytotoxic fraction against T47D cells, we aimed to isolate the cytotoxic compounds and further elucidate their apoptotic mechanisms. Methods:The compounds were isolated through a series of chromatography with cytotoxicity evaluations. Identification of the isolated compounds was achieved by intensive spectroscopic analysis such as NMR, MS, and IR spectra. Cytotoxicity was evaluated by MTT method using doxorubicin as a reference compound. The expression of apoptosis-related factors was quantified by flow cytometry and immunocytochemistry. Results:Two isomers of pregnane steroids with molecular weight 330.2087 (C21H30O3) were isolated from the EtOAc extract. Spectroscopic analysis revealed the structures as 17-ethylene-3,4-dihydroxy-14-methyl-18-norandrostene-16-one (1) and 17-ethylene-3,4-dihydroxy-5-pregnene-16-one (2), respectively. These compounds showed moderate cytotoxicity (IC50 172.9 and 62.2 µg/mL, respectively) comparable to doxorubicin (IC50 3.08 µg/mL). The execution of apoptosis may be related to the increase of the ratio of BAX/bcl-2 of the cells. Conclusion:The EtOAc fraction of Melia azedarach L. leaves and the isolated 5-pregnene-16-one steroids are promising reagents for breast cancer treatment by introducing apoptosis to tumor cells. However, further researches are required to highlight its safety and usage in vivo.

Development of Simultaneous Determination Method of a Pregnan Steroid with Gestagenic Activity – Gestobutanoil and Two its Metabolites in Rat Serum

Introduction. Gestobutanoil is a synthetic pregnane steroid with gestagenic activity. Gestobutanoil has two pharmacologically active metabolites (AMOL and megestrol acetate). This implies the need for a detailed study of the kinetics of metabolites. It is rational to combine the study of the pharmacokinetics of gestobutanoil and its metabolites (AMOL and megestrol acetate). The simultaneous determination of several analytes in the rats’ serum can be carried out using chromatography-mass-spectrometry.Aim. Development of an analytical method for the simultaneous determination of gestobutanoil and two its metabolites in a biomatrix (rat serum).Materials and methods. The following methods were used to determine gestobutanoyl and two its metabolites in a biological matrix: GC-MS, HPLCESI-MS, HPLC-ESI-MS with derivatization, HPLC-APCI-MS.Results and discussion. When working with GC-MS, the chromatographic peaks of gestobutanoyl, AMOL, and megestrol acetate were strongly blurred and superimposed on each other, which is apparently due to the thermolability of the substances. The GC-MS method was abandoned in favor of HPLC. Analytes were separated by HPLC gradient elution on a C18 column. ESI ionization did not give typical protonated ions of gestobutanoyl and AMOL, and the intense signals of their cationized ions and fragment ions, which were observed in the spectra of AMOL and gestobutanoyl, could not ensure the reproducibility of the spectra, since the conditions of their formation are not suitable for routine analysis. Derivatization of analytes to form oximes and substituted hydrazones did not give the expected reaction products for HPLC-ESI-MS. APCI made it possible to remove intense cationized ions from the spectra of gestobutanoyl and AMOL and to increase the reliability of the method. The HPLCAPCI-MS technique was reproduced on model rat blood serum.Conclusion. An HPLC-MS method was developed for the simultaneous determination of gestobutanoyl, megestrol acetate, and AMOL. The technique was tested on a model rat blood serum containing all three analytes.

Structurally diverse steroids with nitric oxide inhibitory activities from Aglaia lawii leaves

Eleven previously uncharacterized steroids, along with three analogs were isolated from Aglaia lawii leaves. Their structures were definitely characterized by the methods of NMR, MS, IR, ECD and X-ray crystallography study. Among these unreported compounds, 3-epi-dyscusin C, 3-epi-lansisterone E and (Z)-2α-hydroxyaglawone were C-21 pregnane steroids incorporating a highly oxygenated ring A, while others were Δ5-3β-hydroxy-7-ketosteroids bearing different ring D and C-17 aliphatic chains. All isolates were evaluated for nitric oxide (NO) inhibitory activities. 3-Epi-dyscusin C, 3-epi-lansisterone E, (Z)-2α-hydroxyaglawone and 17(20)E-dyscusin B showed significant anti-inflammatory activities with IC50 values of NO inhibition less than 10 μM (in the range from 4.47 ± 0.36 to 7.67 ± 0.46 μM).

Four New Pregnane-10,2-carbolactones from an Epipolasis sp. Marine Sponge.

Four new pregnane steroids, 3β,4β,16β-trihydroxypregna-5,17-diene-10,2-carbolactone (1), 16β-acetoxy-3β,4β-dihydroxypregna-5,17-diene-10,2-carbolactone (2), 12β-acetoxy-3β,4β,16β-trihydroxypregna-5,17-diene-10,2-carbolactone (3), and 12β,16β-diacetoxy-3β,4β-dihydroxypregna-5,17-diene-10,2-carbolactone (4) were isolated from an extract of an Epipolasis sp. marine sponge. The structures of the new compounds were determined by extensive NMR spectroscopic analysis and comparison with data from previously reported compounds.

Hydroxy Derivatives of Poststerone and Its Nontrivial 13(14→8)-Abeo-analogues: Synthesis, Crystal Packing, and Intermolecular Hydrogen Bonds

ABSTRACT The 6,20-dioxo-Δ2,3-pregnane and 6,20,14-trioxo-13(14→8)-abeo-pregnane steroid hydroxyl derivatives are synthesized via the reduction with metal hydrides (NaBH4, LiAlH4, and L-Selecride). The structures of the derivatives were elucidated with single-crystal X-ray diffraction analysis, MALDI TOF/TOF mass spectrometry, and 1D and 2D NMR spectroscopy. The topological analysis with a QTAIM approach, the analysis of molecular electrostatic potentials mapped on a Hirshfeld surface, and the CLP-PIXEL method show unequivocally that there is a complex network of intramolecular and intermolecular interactions in the crystals of the pregnane steroids. The molecules of 20(R)-2β,3β-dimesyloxy-14α,20-dihydroxy-5β-pregn-7-en-6-one 2 form a 2D motif (network) due to strong O–H...O hydrogen bonds. The (8R)-8,13-cyclo-13,14-seco-5β-pregn-2-ene-6,14,20-trione 4 and 6α,8R-8,13-cyclo-13,14-seco-5β-pregn-2(3)-en-6-hydroxy-14,20-dione 9 provide two-dimensional network of a brick wall type in their crystals due to weak С–Н...О contacts, whereas 6α,14α,8R,20R-8,13-cyclo-13,14-seco-5β-pregn-2(3)-en-6,14,20-triol 10 forms one-dimensional network (chains) in its crystals owing to strong O–H...O intermolecular interactions. When the number of hydroxyl groups in abeo-steroid increases, the contribution of O...H contacts to the Hirshfeld surface decreases from 26 to 11%.

A Pregnane Steroid as the Chiral Auxiliary in 1,3‐Dipolar Azomethine Ylide's Cycloaddition: Asymmetric Synthesis and Anticancer Activity of Novel Hybrid Compounds

AbstractHybrid chiral molecules containing frameworks of a pregnane steroid and 5‐arylpyrrolidine‐2,4‐dicarboxylate have been synthesized for the first time in a diastereomerically pure form. Chiral pregnane‐based acrylate induced stereoselective formation of all‐cis trisubstituted pyrrolidine moiety under cycloaddition with stabilized N‐metalated azomethine ylides. β‐Dipeptidic fragment was built up at C‐3 pregnane position using consequent acryloylation and stereoselective cycloaddition with stabilized N‐metalated azomethine ylide. All novel hybrid molecules demonstrated micromolar cytotoxic activity against human cervical epithelioid cancer HeLa cell culture in native and estradiol‐stimulated forms and breast carcinoma MCF‐7 cell culture. The most of hybrid compounds are less toxic towards human skin fibroblasts HSF as compared with studied hormonal tumor cells.

Anti-adipogenic Pregnane Steroid from a Hydractinia-associated Fungus, Cladosporium sphaerospermum SW67

A pregnane steroid, 3α-hydroxy-pregn-7-ene-6,20-dione (1), was isolated from a Hydractiniaassociated Cladosporium sphaerospermum SW67 by repetitive column chromatographic separation and highperformance liquid chromatography (HPLC) purification. The planar structure of 1 was elucidated from the analysis of the spectroscopic data (1D and 2D NMR spectra) and LC-MS data. The absolute configuration of 1 was determined by interpretation of ROESY spectrum of 1, together with the comparison of reported spectroscopic values in previous studies. To the best of our knowledge, this is the first report of the identification of the pregnane scaffold from C. sphaerospermum, a natural source. Compound 1 was evaluated for its effects on lipid metabolism and adipogenesis during adipocyte maturation and showed that compound 1 substantially inhibited lipid accumulation compared to the control. Consistently, the expression of the adipocyte marker gene (Adipsin) was reduced upon incubation with 1. Further, we evaluated the effects of 1 on lipid metabolism by measuring the transcription of lipolytic and lipogenic genes. The expression of the lipolytic gene ATGL was significantly elevated upon exposure to 1 during adipogenesis, whereas the expression of lipogenic genes FASN and SREBP1 was significantly reduced upon treatment with 1. Thus, our findings provide experimental evidence that the steroid derived from Hydractinia-associated C. sphaerospermum SW67 is a potential therapeutic agent for obesity.

Pregnane steroidogenesis is altered by HIV-1 Tat and morphine: Physiological allopregnanolone is protective against neurotoxic and psychomotor effects

Pregnane steroids, particularly allopregnanolone (AlloP), are neuroprotective in response to central insult. While unexplored in vivo, AlloP may confer protection against the neurological dysfunction associated with human immunodeficiency virus type 1 (HIV-1). The HIV-1 regulatory protein, trans-activator of transcription (Tat), is neurotoxic and its expression in mice increases anxiety-like behavior; an effect that can be ameliorated by progesterone, but not when 5α-reduction is blocked. Given that Tat's neurotoxic effects involve mitochondrial dysfunction and can be worsened with opioid exposure, we hypothesized that Tat and/or combined morphine would perturb steroidogenesis in mice, promoting neuronal death, and that exogenous AlloP would rescue these effects. Like other models of neural injury, conditionally inducing HIV-1 Tat in transgenic mice significantly increased the central synthesis of pregnenolone and progesterone's 5α-reduced metabolites, including AlloP, while decreasing central deoxycorticosterone (independent of changes in plasma). Morphine significantly increased brain and plasma concentrations of several steroids (including progesterone, deoxycorticosterone, corticosterone, and their metabolites) likely via activation of the hypothalamic-pituitary-adrenal stress axis. Tat, but not morphine, caused glucocorticoid resistance in primary splenocytes. In neurons, Tat depolarized mitochondrial membrane potential and increased cell death. Physiological concentrations of AlloP (0.1, 1, or 10 nM) reversed these effects. High-concentration AlloP (100 nM) was neurotoxic in combination with morphine. Tat induction in transgenic mice potentiated the psychomotor effects of acute morphine, while exogenous AlloP (1.0 mg/kg, but not 0.5 mg/kg) was ameliorative. Data demonstrate that steroidogenesis is altered by HIV-1 Tat or morphine and that physiological AlloP attenuates resulting neurotoxic and psychomotor effects.