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The genus Polygonatum Mill. belongs to the Liliaceae family, which is widely distributed all over the world. Modern studies have found that Polygonatum plants are very rich in chemical compounds such as saponins, polysaccharides and flavonoids. Steroidal saponins are the most commonly studied saponins in the genus Polygonatum and a total of 156 compounds have been isolated from 10 species of the genus. These molecules possess antitumor, immunoregulatory, anti-inflammatory, antibacterial, antiviral, hypoglycemic, lipid-lowering and anti-osteoporotic activities. In this review, we summarize recent advances in studies of the chemical constituents of steroidal saponins from Polygonatum, including their structural characteristics, possible biosynthetic pathways and pharmacological effects. Then, the relationship between the structure and some physiological activities is considered. This review aims to provide reference for further exploitation and utilization of the genus Polygonatum., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

New steroidal 3β,21-disulfates ( 2 - 4 ), steroidal 3β,22-disulfate ( 5 ), and the previously known related steroidal 3β,21-disulfate ( 1 ) were isolated from the ethanolic extract of the Far Eastern starfish Pteraster marsippus , collected off Urup Island in the Sea of Okhotsk. The structures of these compounds were determined by intensive NMR and HRESIMS techniques as well as by chemical transformations. Steroids 2 and 3 have an oxo-group in the tetracyclic nucleus at position C-7 and differ from each other by the presence of the 5(6)-double bond. The Δ 24 -22-sulfoxycholestane side chain of the steroid 5 has not been found previously in the starfish or ophiuroid steroids. The cytotoxic activities of 1 , 4 , 5 , and the mixture of 2 and 3 were determined on the models of 2D and 3D cultures of human epithelial kidney cells (HEK293), melanoma cells (SK-MEL-28), small intestine carcinoma cells (HuTu80), and breast carcinoma cells (ZR-75-1). The mixture of 2 and 3 revealed a significant inhibitory effect on the cell viability of human breast carcinoma ZR-75-1 cells, but other tested compounds were less effective.

Gorgostane steroids are isolated from marine organisms and consist of 30 carbon atoms with a characteristic cyclopropane moiety. From the pioneering results to the end of 2021, isolation, biosynthesis, and structural elucidation using 13 C-NMR will be used. Overall, 75 compounds are categorized into five major groups: gorgost-5-ene, 5,6-epoxygorgostane, 5,6-dihydroxygorgostane, 9,11-secogorgostane, and 23-demethylgorgostane, in addition to miscellaneous gorgostane. The structural diversity, selectivity for marine organisms, and biological effects of gorgostane steroids have generated considerable interest in the field of drug discovery research.

The species of Paris genus is a prolific source of structurally diverse steroidal saponins responsible for multivarious biological properties. The first phytochemical investigation on the steroidal saponin constituents from the rhizomes of Paris vaniotii Lévl. led to the discovery and structural characterization of four new spirostanol saponins, named parisvaniosides A-D (1-4), and one new furostanol glycoside, named parisvanioside E (5), along with eleven known analogues (6-16). Their structures were unambiguously established on the basis of extensive spectroscopic analysis and comparison with the reported spectroscopic data. Compound 1 is a rare spirostanol saponin sharing with a C-9/C-11 double bond and a peroxy group located between C-5 and C-8 of the aglycone, whereas 3 and 4 are unusual C-27 steroidal sapoins with hydroxyl/methoxyl at both C-5 and C-6. Furthermore, 5 is the first furostanol saponin with a unique aglycone featuring two trisubstituted double bonds in ring B. All isolated saponins were evaluated for their anti-inflammatory effects on a lipopolysaccharide (LPS)-stimulated nitric oxide (NO) production model in RAW 264.7 macrophages., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Trillium govanianum , commonly known as Nag Chhatri and Teen Patra, is a popular herbal supplement traditionally used for curing different inflammatory and sexual disorders, infection and wound healing. Steroidal saponins are considered as active components of this species. The present study demonstrated the isolation of nine steroidal saponins, including one new compound named as govanoside B ( 9 ) and eight known, pregna-chacotrioside ( 1 ), pennogenin-triglycoside ( 2 ), borassoside E ( 3 ), pennogenin-tetraglycoside ( 4 ), protodioscin ( 5 ), clintonioside B ( 6 ), pennogenin-diglycoside ( 7 ) and borassoside D ( 8 ). This is the first report on the isolation of 1 , 2 , 4 , 5 , 6 , 7 and 8 from rhizomes of T. govanianum . The extract, fractions and isolated compounds were further evaluated for their DPPH and ABTS radical scavenging activity.

Context: Ircinia mutans Wilson (Irciniidae) is a sponge with antimicrobial and cytotoxic constituents., Objective: Our objective was to characterise the cytotoxic constituents of two seasonal collections of I. mutans ., Materials and Methods: The sponges were extracted in methanol-dichloromethane and their constituents were purified and characterised using column chromatography, GC-MS, 1 D and 2 D NMR. Anti-proliferative activities of the compounds, were evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay (0.25-100 μg/mL, 72 h) against leukaemia (MOLT-4), breast (MCF-7) and colon cancer (HT-29) human cells., Results: Three furanosesquiterpoids; furodysin ( 1 ), ent -furodysinin ( 2 ) and furoircin ( 3 ) and ten sterols were characterised in I. mutans , for the first time. Cholesterol ( 4 ), cholesta-5, 7-dien-3β-ol ( 5 ) and ergosterol ( 6 ) were determined in the sponge from the winter collections, while cholesta-5, 22-dien-3β-ol ( 7 ), 24-methyldesmosterol ( 8 ), campesterol ( 9 ), stigmasterol ( 10 ), γ-ergostenol ( 11 ), chondrillasterol ( 12 ) and γ-sitosterol ( 13 ) were detected in the summer samples. The steroids from the winter collection exhibited cytotoxic activity with IC 50 values of 13.0 ± 0.9, 11.1 ± 1.7 and 1.1 ± 0.4 µg/mL, against the mentioned cancer cell lines, respectively, while those from the summer sample, showed greater activity, IC 50 = 1.1 ± 0.2 μg/mL against MOLT-4. The purified steroids showed potent MOLT-4 cytotoxic activity, IC 50 values = 2.3-7.8 µg/mL., Discussion and Conclusion: The present study suggests that I. mutans is a rich source of cytotoxic steroids, and introduces 3 as new natural product. Considering the high cytotoxic activity of the steroids, these structures could be candidates for anticancer drug development in future research.

Fourteen polyhydroxylated steroids including six new compounds, ehrensteroids A - F (1-6), were isolated from the Vietnamese soft coral Sarcophyton ehrenbergi after applying various chromatographic separations. The structure elucidation was done by detailed analysis of 1D, 2D NMR and HR QTOF mass spectra. In addition, significant cytotoxicity (IC 50 values ranging from 8.45 ± 0.56 to 38.27 ± 2.39 μM) against all five tested cancer cell lines as LU-1 (lung cancer), HepG2 (hepatoma cancer), MCF-7 (breast cancer), HL-60 (acute leukemia), and SW480 (colon adenocarcinoma) was observed for compounds 1, 2, 6-10, and 12-14, whereas this activity on LU-1 (IC 50  = 21.91 ± 2.72 μM) and HepG2 (IC 50  = 20.32 ± 0.03 μM) cell lines was also observed for 4., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Gymnema sylvestre (Retz.) Schult is a multi-purpose traditional medicine that has long been used for the treatment of various diseases. To discover the potential bioactive composition of G. sylvestre , a chemical investigation was thus performed. In this research, four new C 21 steroidal glycosides sylvepregosides A-D ( 1 - 4 ) were isolated along with four known compounds, gymnepregoside H ( 5 ), deacetylkidjoladinin ( 6 ), gymnepregoside G ( 7 ) and gymnepregoside I ( 8 ), from the ethyl acetate fraction of G. sylvestre . The structures of the new compounds were established by extensive 1D and 2D nuclear magnetic resonance (NMR) spectra with mass spectroscopy data. Compounds 1 - 6 promoted glucose uptake by the range of 1.10- to 2.37-fold, respectively. Compound 1 showed the most potent glucose uptake, with 1.37-fold enhancement. Further study showed that compounds 1 and 5 could promote GLUT-4 fusion with the plasma membrane in L6 cells. The result attained in this study indicated that the separation and characterization of these compounds play an important role in the research and development of new anti-diabetic drugs and pharmaceutical industry.

Chemical investigation of the marine sponge Dysidea avara, collected from the South China Sea, yielded 13 steroids, including nine new (1-9) and four known (10-13) ones. The new structures were elucidated as (3S,14R)-3,14-dihydroxycholesta-5,8-dien-7-one (1), (22E,24R)-7α-ethoxy-5α,6α-epoxyergosta-8(14),22-dien-3β-ol (2), 3β-hydroxy-7α-ethoxy-5α,6α-epoxy-8(14)-cholestene (3), 3β,5α-dihydroxy-6α-ethoxychofesta-7,9(11)-diene (4), 3β,5α-dihydroxy-6β-ethoxycholest-7-ene (5), (22E,24R)-24-ethoxy-3β,5α-dihydroxy-6β-ethoxyergosta-7,22-diene (6), (22E)-3β,5α-dihydroxy-6β-ethoxycholesta-7,22-diene (7), 24-ethoxy-3β,5α-dihydroxy-6β-ethoxycholest-7-ene (8 and 9), by extensive spectroscopic analyses, such as HR-ESI-MS, 1D and 2D NMR data. The absolute configuration of 1 was assigned by comparison the experimental ECD spectra with the calculated ones. Among the 13 metabolites, compounds 1, 4, 11, 12, and 13 showed NF-κB inhibitory activities in human HER-293 cells with IC 50 values of 6.4, 18.7, 8.1, 9.6, and 7.5 μM, respectively. Preliminary structure-activity relationship analysis unveiled that the conjugated ketones or unsaturated double bonds might be the functional groups for the five active steroids., (© 2021 Wiley-VHCA AG, Zurich, Switzerland.)

In the ongoing research on potent antitumor agents from the rhizomes of Asparagus cochinchinensis, seven undescribed steroidal saponins asparagusoside A-G (1-7), along with twenty known ones (8-27), were isolated and elucidated via analyzing their 1D, 2D NMR, mass spectroscopic data and chemical methods. All isolated compounds were evaluated for their cytotoxic effects against human large cell lung carcinoma cells (NCI-H460) in vitro. Among them, compounds 7, 9 and 27 showed more significant antitumor activities than the positive control cisplatin (11.56 μM) with IC 50 values of 1.39, 3.04, and 2.25 μM, respectively. Further research about asparagusoside G (7) showed G0/G1 arrest in NCI-H460 cell line cycle and induced cell death by apoptosis in a dose‑dependent way., (Copyright © 2021 Elsevier Inc. All rights reserved.)

A phytochemical study on the fruits of Artocarpus heterophyllus caused the isolation of a previously undescribed steroid, artoheterophoid ( 1 ), together with seven known analogues ( 2 - 8 ). The chemical structure of 1 was elucidated on the basis of comprehensive spectroscopic methods and the known compounds ( 2 - 8 ) were identified by comparing their spectroscopic data with those reported in the literatures. All known compounds ( 2 - 8 ) were separated from A. heterophyllus for the first time. All isolated compounds ( 1 - 8 ) were assessed for their anti-inflammatory activities in vitro by measuring the inhibitory effect against nitric oxide (NO) production induced by lipopolysaccharide in mouse macrophage RAW 264.7 cells. Compounds 1 - 8 displayed remarkable inhibitory effects against NO production with the IC 50 values in the range of 0.72 ± 0.07 to 5.93 ± 0.12 μM.

Seven highly oxidized steroids, taccachatrones A-G ( 1 - 7 ), together with four known taccalonolides ( 8 - 11 ), were characterized from the rhizomes of Tacca chantrieri . The structures of 1 - 7 were established on the basis of spectroscopic data analysis, while the absolute configurations were determined by single-crystal X-ray diffraction. Compounds 1 - 4 may be derived from taccalonolide derivatives by the degradation of three carbon atoms. Compounds 7 , 8 , 10 , and 11 exhibited cytotoxicity to human cancer cell lines, indicating that the presence of a lactone moiety, as well as a double bond between C-22 and C-23, might play key roles in mediating their cytotoxicity.

Three pregnene steroids, two of them new, were isolated from ethyl-acetate partition of liquid-cultivation of the extremophyle fungus Exophiala oligosperma , found in a pH 1.5 hydrochloric acid aqueous solution. Spectroscopic studies using NMR and HRMS, allowed the identification of the molecular structures of both Δ 8,9 -pregnenes, still not described in the literature.

A new withanolide glycoside ( 1 ), two new ergostanol glycosides ( 2 and 3 ), and a new furostanol glycoside ( 4 ), along with nine known steroidal derivatives ( 5 - 12 ) were isolated from the seeds of Withania somnifera . The structures of the new compounds were determined using spectroscopic analysis and hydrolysis. The cytotoxic activities of the isolated compounds were evaluated against Ca9-22 human gingival carcinoma cells, HSC-2 human mouth carcinoma cells, and HL-60 human promyelocytic leukemia cells. Only 12 exhibited cytotoxic activity against these cell lines with IC 50 values of 0.38, 0.54, and 1.5 μM, respectively.

Two new steroidal glycosides oxystauntoside A (1) and oxystauntoside B (2), together with sixteen known compounds (3-18) were isolated from the 95% ethanol extract of Merrillanthus hainanensis. Their structures were characterized by extensive spectroscopic analysis including NMR and mass spectra and single crystal X-ray crystallography. The absolute configuration of 1 and 2 were further determined by ECD calculations. All of these compounds were isolated from M. hainanensis for the first time. All the fractions and compounds were tested for the anti-inflammatory activity against the TNF-α factor. The ethyl acetate fraction showed the most potent inhibition (71.3%) at 10 μg/mL and compounds 5 (78.9%) and 9 (73.4%) in this fraction with both carboxyl and phenolic hydroxyl groups showed significant inhibition at 10 μM. Our study provided the first scientific report for the medicinal value of M. hainanensis., (Copyright © 2021. Published by Elsevier B.V.)

Chemical investigation on the ethyl acetate extract of the aerial parts of Begonia sp. afforded a new steroid glycoside, 9(11)α,16(17)α-dioxirane-20,25-dihydroxy-β-sitosterol-3- O -β-glucopyranoside ( 1 ) along with a known steroidal glycoside, β-sitosterol-3- O -β-D-glucopyranoside ( 2 ). The Chemical structures were elucidated by 1D and 2D NMR and mass spectroscopic analysis. Cytotoxicity against four different cancer cell lines (HeLa, T47D, WiDr and Vero) was assessed. Compound 1 was more potent and selective against breast cancer cell line (T47D) than other cell lines with an IC 50 value of 0.16 µg/mL. Further docking study of 1 exhibited the preference of molecule to bind in the epidermal growth factor receptor tyrosine kinase (EGFR-TK) binding pockets with docking scores of -97.8800 (PLANTS) and -3.56 kcal/mol (AutoDock 4.2.6).

Steroidal alkaloids (1-11), including one new 24-hydroxylated cevanine-type steroidal alkaloid, named yibeinone F (1), were isolated from the bulbs of Fritillaria pallidiflora Schrenk. Their structures were elucidated by analyses of extensive spectroscopic data and comparison of the NMR data with those reported previously, and the structures of compounds 1, 7 and 11 were further confirmed by X-ray single crystal diffraction analyses. The anti-inflammatory effects of all the isolated alkaloids were evaluated in LPS-activated RAW264.7 macrophages. Among them, compounds 9 (stenanzine) and 10 (hapepunine) showed significant inhibitory effects against LPS-induced NO production with IC 50 values of 8.04 μM and 20.85 μM, respectively. Furthermore, compound 9 effectively inhibited the release of cytokines such as interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and prostaglandin E2 (PGE 2 ), and suppressed the protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX2) in LPS-stimulated RAW264.7 cells. Further experiments revealed the underlying mechanism that 9 blocked LPS-induced phosphorylation and degradation of inhibitor-α of nuclear transcription factor κB (IκBα) and c-Jun N-terminal kinase (JNK) in RAW264.7 cells. Taken together, compound 9 may be a valuable candidate for the treatment of inflammatory diseases., (Copyright © 2021 Elsevier Inc. All rights reserved.)

As our ongoing chemical investigation, two new pregnane steroidal glycosides, cynataihosides G ( 1 ), with a new aglycone, and H ( 2 ) were isolated from the 95% ethanol extract of Cynanchum taihangense . Their structures were elucidated on the basis of 1 D and 2 D NMR spectral data, HR-ESI-MS analysis and qualitative chemical methods. The compounds were subjected to detect the cytotoxicity against three human tumor cell lines (HL-60, THP-1 and PC-3). The compounds displayed no significant cytotoxicity.Supplemental data for this article can be accessed at https://doi.org/10.1080/14786419.2019.1672682.

A chemical investigation on the EtOAc extract of the endophytic fungus Eutypella scoparia SCBG-8 led to the isolation of eight new sesquiterpenes eutyscoparins A-H ( 1-8 ), one C-28 steroid eutyscoparene A ( 9 ), one triterpenoid eutyscoparene B ( 10 ), six known terpenoids, and two known steroids. Their structures including absolute configurations were established on the basis of spectroscopic data analysis, single-crystal X-ray diffraction experiments, and electronic circular dichroism (ECD) calculations. Compound 7 displayed antibacterial activity against S. aureus and MRSA (methicillin-resistant Staphylococcus aureus ) with MIC values of 6.3 μg/mL.

Sinularia is one of the conspicuous soft coral species widely distributed in the world's oceans at a depth of about 12 m. Secondary metabolites from the genus Sinularia show great chemical diversity. More than 700 secondary metabolites have been reported to date, including terpenoids, norterpenoids, steroids/steroidal glycosides, and other types. They showed a broad range of potent biological activities. There were detailed reviews on the terpenoids from Sinularia in 2013, and now, it still plays a vital role in the innovation of lead compounds for drug development. The structures, names, and pharmacological activities of compounds isolated from the genus Sinularia from 2013 to March 2021 are summarized in this review.

Henryiosides F and G ( 1 and 2 ), two new steroidal saponins along with two known analogues ( 3 and 4 ) were obtained from the roots and rhizomes of Maianthemum henryi . Their structures were determined by physicochemical properties and spectroscopic methods including 1D, 2D-NMR, IR and HR-ESI-MS data analysis. Cytotoxic activity in human HepG2 and SW620 tumour cells were evaluated by the MTT method and all of the saponins exhibited cytotoxicity with IC 50 values ranging from 15.33 μ M to 57.85 μ M.

The review focuses on sulfated steroids that have been isolated from seaweeds, marine sponges, soft corals, ascidians, starfish, and other marine invertebrates. Sulfur-containing steroids and triterpenoids are sourced from sedentary marine coelenterates, plants, marine sediments, crude oil, and other geological deposits. The review presents the pharmacological profile of sulfated steroids, sulfur-containing steroids, and triterpenoids, which is based on data obtained using the PASS program. In addition, several semi-synthetic and synthetic epithio steroids, which represent a rare group of bioactive lipids that have not yet been found in nature, but possess a high level of antitumor activity, were included in this review for the comparative pharmacological characterization of this class of compounds. About 140 steroids and triterpenoids are presented in this review, which demonstrate a wide range of biological activities. Therefore, out of 71 sulfated steroids, thirteen show strong antitumor activity with a confidence level of more than 90%, out of 50 sulfur-containing steroids, only four show strong antitumor activity with a confidence level of more than 93%, and out of eighteen epithio steroids, thirteen steroids show strong antitumor activity with a confidence level of 91% to 97.4%.

Vernoramyosides A-F (1-6), six new Δ 7,9(11) stigmastane-type steroid saponins, along with four known analogues (7-10) were isolated from the leaves of Vernonia amygdalina Delile (Compositae). Their structures were determined by the combination of NMR, ECD and HR-ESI-MS data. These compounds all possessed highly oxidized side chain and a γ-lactam or α,β-unsaturated five-membered lactone ring. All isolates were screened for their activities in reversing resistance in MCF/DOX cells., (Copyright © 2021 Elsevier B.V. All rights reserved.)

A new coprostanol glycoside, 26-trihydroxy-16, 22-dioxo-3 β -[( α -L-rhamnopyranosyl)oxy]-5 β -cholestan-1 β -yl O - α -L-rhamnopyranosyl-(1→2)- β -D-fucopyranoside ( 1 ) (25 S )-5 β -spirostan-1 β ,2 β ,3 β , 4 β ,5 β , 6 β -hexol ( 2 ), a new C-22 steroidal lactone saponin, (20  R )-16 β -trihydroxy-3 β -[( α -L-rhamnopyranosyl)oxy]-1 β -[( O - α -L-rhamnopyranosyl-(1→2)- β -D-xylopyranosyl)oxy]-5 β -pregnane-20-oic acid γ-lactone ( 3 ) along with two known pregnane glycosides ( 4 and 5 ) were obtained from the roots of Reineckia carnea . Their structures were determined by 1 D, 2 D NMR, IR and MS data analysis. In addition, the cytotoxic activities in HT-29, HCT116, H1299 and A549 tumor cells of the isolated compounds ( 1  -  5 ) were determined by the MTT method. The results showed that only compound 1 exhibited weak effect against these cells with IC 50 values ranging from 63.36  μ M to 105.01  μ M.

Phellinus Quél is one of the largest genera of Hymenochaetaceae, which is comprised of about 220 species. Most Phellinus macro-fungi are perennial lignicolous mushrooms, which are widely distributed on Earth. Some Phellinus fungi are historically recorded as traditional medicines used to treat various diseases in eastern Asian countries, especially China, Japan and Korean. Previous phytochemical studies have revealed that Phellinus fungi produce diverse secondary metabolites, which mainly contain polysaccharides, flavones, coumarins, terpenes, steroids, and styrylpyranones. Pharmacological documents have demonstrated that Phellinus mushrooms and their compounds have a variety of bioactivities, such as anti-tumor, immunomodulation, anti-oxidative and anti-inflammation, anti-diabetes, neuro-protection, and anti-viral effects. This review surveys the literature reporting the isolation, characterization, and bioactivities of secondary metabolites from the fungi of the genus Phellinus, focusing on studies published in the literature up to April 2020. Herein, a total of more than 300 compounds from 13 Phellinus species and their isolation, characterization, chemistry, pharmacological activities, and relevant molecular mechanisms are comprehensively summarized.

Naturally occurring C 21 -steroidal aglycones from Cynanchum exhibit significant antitumor effects. To expand the chemical diversity and get large scale C 21 -steroidal aglycones, the extracts of the roots of Cynanchum otophyllum were treated with 5% HCl in aqueous and the resulting hydrolysate was investigated. Nine new C 21 -steroidal aglycones (1-9) namely cynotogenins A-I, along with seventeen known analogous (10-26), were isolated from the hydrolysate. The structures of compounds 1-9 were elucidated by spectroscopic analysis (IR, HR-ESI-MS, 1D and 2D NMR) and comparison of observed spectroscopic data with those of reported in the literature. Aglycones 2-5 with rare cis-cinnamoyl group as well as 8 and 9 with 5β,6β-epoxy group were found from the genus of Cynanchum for the first time. The cytotoxicities of compounds 1-26 toward human cancer HeLa, H1299, HepG2, and MCF-7 cells were evaluated and preliminary structure-activity relationship (SAR) was discussed. Moreover, compound 20 inhibits HepG2 cell apoptosis and induces of G0/G1 phase arrest in a dose dependent manner., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Two new polyketides, xerucitrinin A ( 1 ) and coniochaetone M ( 8 ), and one new steroid, 16 α -methylpregna-17 α ,19-dihydroxy-(9,11)-epoxy-4-ene-3,18-dione-20-acetoxy ( 13 ), together with eleven known analogues were isolated from fungus Penicillium citrinum SCSIO 41017 associated with the sponge Callyspongia sp. Their structures and absolute configurations were elucidated by NMR spectra, MS, CD, optical rotation, X-ray crystallography, and compared with literature data. Biological evaluation results revealed that 5 exhibited significant cytotoxic activity against MCF-7 cell line with IC 50 values of 1.3  μ M. Compound 13 showed moderate activity against all cell lines with IC 50 values of 13.5-18.0  μ M, and compounds 9 and 14 showed weak activity with MIC values of ranging from 125  μ g/mL to 250  μ g/mL respectively.[Formula: see text].

For several decades now, the analysis of steroids has been a key tool in the diagnosis and monitoring of numerous endocrine pathologies. Thus, the available methods used to analyze steroids in biological samples have dramatically evolved over time following the rapid pace of technology and scientific knowledge. This review aims to synthetize the advances in steroids' analysis, from classical approaches considering only a few steroids or a limited number of steroid ratios, up to the new steroid profiling strategies (steroidomics) monitoring large sets of steroids in biological matrices. In this context, the use of liquid chromatography coupled to mass spectrometry has emerged as the technique of choice for the simultaneous determination of a high number of steroids, including phase II metabolites, due to its sensitivity and robustness. However, the large dynamic range to be covered, the low natural abundance of some key steroids, the selectivity of the analytical methods, the extraction protocols, and the steroid ionization remain some of the current challenges in steroid analysis. This review provides an overview of the different analytical workflows available depending on the number of steroids under study. Special emphasis is given to sample treatment, acquisition strategy, data processing, steroid identification and quantification using LC-MS approaches. This work also outlines how the availability of steroid standards, the need for complementary analytical strategies and the improvement of calibration approaches are crucial for achieving complete steroidome quantification., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Six new ( 1 , 2 , 6 , 8 , 13, and 20 ) and twenty previously isolated ( 3 - 5 , 7 , 9 - 12 , 14 - 19 , and 21 - 26 ) steroids featuring thirteen different carbocycle motifs were isolated from the organic extract of the soft coral Sinularia polydactyla collected from the Hurghada reef in the Red Sea. The structures and the relative configurations of the isolated natural products have been determined based on extensive analysis of their NMR and MS data. The cytotoxic, anti-inflammatory, anti-angiogenic, and neuroprotective activity of compounds 3 - 7 , 9 - 12 , 14 - 20 , and 22 - 26 , as well as their effect on androgen receptor-regulated transcription was evaluated in vitro in human tumor and non-cancerous cells. Steroids 22 and 23 showed significant cytotoxicity in the low micromolar range against the HeLa and MCF7 cancer cell lines, while migration of endothelial cells was inhibited by compounds 11 , 12 , 22, and 23 at 20 µM. The results of the androgen receptor (AR) reporter assay showed that compound 11 exhibited the strongest inhibition of AR at 10 µM, while it is noteworthy that steroids 10 , 16, and 20 displayed increased inhibition of AR with decreasing concentrations. Additionally, compounds 11 and 23 showed neuroprotective activity on neuron-like SH-SY5Y cells.

As part of our current work to discover structurally and/or biologically novel compounds from Korean wild mushrooms, we isolated five ergostane-type steroids (1-5) from the fruiting bodies of Xerula furfuracea via repeated column chromatographic separations and HPLC purification. The chemical structures of the isolated steroids were shown to be (22E,24 R )-24-methylcholesta-4,22- diene-3,6-dione (1), ergosta-7,22-diene-3 β ,5 α ,6 β -triol (2), ergosta-7,22-diene-3 β ,5 α ,6 β ,9 α -tetraol (3), (22E,24 R )-5 α ,8 α -epidioxyergosta-6,22-diene-3 β -ol-3-O- β -D-glucopyranoside (4), and (22E,24 R )-5 α ,8 α -epidioxyergosta-6,9,22-triene-3 β -ol-3-O- β -D-glucopyranoside (5) based on comparison of the data regarding their spectroscopic and physical properties with those of previous studies. Notably, this is the first report on the presence of the identified steroids (1-5) in this mushroom. We tested compounds 1-5 to determine their effects on adipogenesis and osteogenesis in the mouse mesenchymal stem cell line C3H10T1/2 and found that compounds 4 and 5 suppressed the differentiation of stem cells into adipocytes. Notably, in addition to its suppressive effect on adipogenesis, compound 5 was also shown to promote the osteogenic differentiation of stem cells. These findings demonstrate that the bioactive compounds isolated might be effective for the treatment of menopause-associated syndromes, such as osteoporosis and obesity, as the isolated compounds were shown to suppress adipogenesis and/or promote osteogenesis of stem cells.

A systematic phytochemical study on the components in the seeds of Allium tuberosum was performed, leading to the isolation of 27 steroidal glycosides (SGs 1 - 27 ). The structures of SGs were identified mainly by nuclear magnetic resonance and mass spectrometries as well as the necessary chemical evidence. In the SGs, 1 - 10 and 22 - 26 are new steroidal saponin analogues. An in vitro bioassay indicates that 1 , 2 , 7 , 8 , 10 , 13 - 15 , 20 , 23 , and 26 display promotional roles in testosterone production of rat Leydig cells with the EC 50 values of 1.0 to 4.5 μM, respectively.

Four new polyhydroxylated steroids plaksterols A-D (1-4), together with two known related steroids ergost-7,9(11),22-trien-3β,5α,6α-triol (5) and ergosta-6β-methoxy-7,22-diene-3β,5α-diol (6), were isolated from methanol extract of the South China Sea marine sponge Plakortis sp. Their structures were identified by spectroscopic analysis, including NMR, MS, and IR. The cytotoxicity of the polyhydroxylated steroids were evaluated, and compound 6 showed moderate inhibitory activities against K562, HL-60 and BEL-7402 cells., (Copyright © 2020 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.)

Two new steroidal alkaloids (1-2), together with seven known related steroidal alkaloids (3-9), were isolated from the rhizomes of Veratrum nigrum L. Their structures were elucidated by extensive spectroscopic analysis, and by comparison with literature data. Compound 1 possessed a rare 1, 3-oxazolidine unit within varazine-type alkaloids, and 2 was a 9-hydroxy-4-one derivative of 3-veratroylgermine. All isolates were evaluated inhibit tomato yellow leaf curl virus (TYLCV) activity. Compounds 5 and 7 (40 μg/mL) showed a significant anti-TYLCV activity in the host Nicotiana benthamiana with inhibition rates 74.6% and 63.4%, respectively, which are higher than that of the positive control ningnanmycin (51.4%)., (Copyright © 2020. Published by Elsevier B.V.)

Four previously undescribed steroids, identified as (3S,7S,8S,9S,10R,13S,14S,16S,17R,20S)-7α-methoxy-ergosta-5,24(28)-dien-3β,16β,20-triol (1), ergosta-5,24(28)-dien-3β,7α,16β-triol (2), ergosta-5,25-dien-3β,7α,16β,20-tetrol (3) and 7α,16β,24α-trihydroxy-varninasterol (4), as well as five known analogues (5-9), were isolated from the leaves and twigs of Dysoxylum pallens Hiern (Meliaceae). Their structures were elucidated based on extensive spectroscopic analysis such as HR-ESI-MS, 1D and 2D NMR, UV, and IR. The absolute configuration of compound 1 was determined by X-ray diffraction analysis. Selected compounds were evaluated for their cytotoxic activities. Compounds 1, 2, and 8 exhibited moderate cytotoxic activity against HL-60, Hela, and HepG2 tumor cell lines with IC 50 ranged from 11.09 to 17.51 μM., Competing Interests: Declaration of Competing Interest All authors involved have no commercial association or other arrangement that might pose or imply a conflict of interest in connection with the submitted article., (Copyright © 2020. Published by Elsevier B.V.)

Hypholoma lateritium is an edible macrofungus with a common distribution in Europe, North America, and the Far East. The aim of this study was to investigate the potential anti-inflammatory effects of H. lateritium extracts and its isolated steroids: fasciculic acid B, fasciculol E, fasciculol C, lanosta-7,9(11)-diene-12β,21α-epoxy-2α,3β,24β,25-tetraol, fasciculol F, and demethylincisterol A2. Organic (hexane, chloroform and 50 % methanol) and water extracts of H. lateritium were subjected to in vitro assays to determine pro-inflammatory protein levels, such as cyclooxygenase-2 (COX-2), cytosolic prostaglandin E2 synthase (cPGES), and antioxidant nuclear factor (erythroid-derived 2)-like 2 (Nrf2). Fungal extracts demonstrated significant activities on pro-inflammatory protein levels with minor differences among the activities of the fractions of different polarities. All the compounds proved to exert notable inhibitory properties on COX-2 and were capable to stimulate the Nrf2 pathway. Fungal extracts and the compounds exerted no cytotoxic activities on RAW 264.7 cells., (© 2020 The Authors. Chemistry & Biodiversity Published by Wiley-VHCA AG, Zurich, Switzerland.)

Based on a method combining the LC-MS/MS molecular networking strategy with the conventional means of phytochemical research, the chemical constituents and the availability of Paris tengchongensis, a new species found in 2017 from Yunnan Province, were investigated for the first time. The molecular networking showed that this species contained the characteristic steroidal glycosides of the genus Paris by comparison of those of Paris polyphylla var. yunnanensis. Furthermore, the detailed investigation on the 80% EtOH extract of its rhizomes resulted to the isolation of twenty steroidal glycosides including three new spirostane-type saponins, named paristengosides A-C (1-3). Their structures were confirmed by spectroscopic analyses (HRMS and NMR) and chemical methods. The new isolates were evaluated for their cytotoxicities against two human cancer cell lines (HEL and MDA-MB-231), anti-inflammatory effects on a lipopolysaccharide (LPS)-stimulated NO production model in RAW264.7 macrophages, anti-AChE, and antimicrobial activities. The results from the molecular networking and the investigation on the chemical constituents suggested that P. tengchongensis can be used as a potential resource of Rhizoma Paridis., Competing Interests: Declaration of Competing Interest The authors declare that they have no financial or non-financial competing interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Seven new polyoxygenated steroids belonging to a new structural group of sponge steroids, gracilosulfates A-G ( 1 - 7 ), possessing 3 β - O -sulfonato, 5 β, 6 β epoxy (or 5(6)-dehydro), and 4 β ,23-dihydroxy substitution patterns as a common structural motif, were isolated from the marine sponge Haliclona gracilis. Their structures were determined by NMR and MS methods. The compounds 1, 2, 4, 6 , and 7 inhibited the expression of prostate-specific antigen (PSA) in 22Rv1 tumor cells.

The simultaneous determination of a large number of steroids (a.k.a. steroid profile) is a powerful tool that provides useful information about the status of steroid hormones. Steroid profile evaluated in matrices such as urine, saliva and plasma provide one-off moment information about the hormonal status and is highly affected by different factors such as circadian rhythm or apprehension to needles. In contrast, the determination of the steroid profile in hair would provide information about the chronic status of the steroid hormones. The objective of the current research was to develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) methodology for the determination of 11 steroids in hair, including 6 hormones and 5 metabolites. We have optimized different parts of the analytical procedure such as (i) hair shredding, (ii) hair amount, (iii) extraction from hair, (iv) extraction time, (v) required extractions and (vi) analytes preconcentration. MS parameters such as the inclusion of ESI- transitions were also evaluated. The optimization of these parameters was found to be critical to achieve the required sensitivity for the determination of steroids in hair. The method was validated with appropriate linearity in the endogenous range, intra- and inter-assay accuracies and matrix effect between 80% and 120% and intra- and inter-assay precisions below 20% for all analytes. Most of the analytes showed to be stable up to 10 months at room temperature. The suitability of the method was evaluated by obtaining the endogenous concentration range of steroids in 30 healthy volunteers. Results agreed with the scarce data previously reported for some steroids. For others, endogenous concentration ranges in hair were reported for the first time. Additionally, the method was used to compare intraindividual levels of steroids in beard and hair. Results revealed that with the exception of testosterone, beard is a suitable alternative to the hair determination of the steroid profile. In summary, the present strategy to evaluate the steroid profile in hair may be a useful tool with a high potential for a wide range of clinical purposes., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Steroid hormones (SH) play a number of important physiological roles in vertebrates including fish. Changes in SH concentration significantly affect reproduction, differentiation, development, or metabolism. The objective of this study was to develop an in vitro high-throughput thin-film solid-phase microextraction (TF-SPME)-liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for targeted analysis of endogenous SH (cortisol, testosterone, progesterone, estrone (E1), 17β-estradiol (E2), and 17α-ethinylestradiol (EE2)) in wild white sucker fish plasma where the concentrations of the analytes are substantially low. A simple TF-SPME method enabled the simultaneous determination of free and total SH concentrations. The use of biocompatible coating allowed direct extraction of these hormones from complex biological samples without prior preparation. The carryover was less than 3%, thereby ensuring reusability of the devices and reproducibility. The results showed that TF-SPME was suitable for the analysis of compounds in the polarity range between 1.28 and 4.31 such as SH at different physicochemical properties. The proposed method was validated according to bioanalytical method validation guidelines. The limit of detection (LOD) and limit of quantification(LOQ) for cortisol, testosterone, progesterone, E1, E2, and EE2 were from 0.006 to 0.150 ng/mL and from 0.020 to 0.500 ng/mL, respectively. The recovery for the method was about 85%, and the accuracy and precision of the method for cortisol, testosterone, and progesterone were ≤ 6.0% and ≤ 11.2%, respectively, whereas those for E1, E2, and EE2 were ≤ 15.0% and ≤ 10.2%, respectively. On the basis of this study, TF-SPME demonstrated several important advantages such as simplicity, sensitivity, and robustness under laboratory conditions. Graphical abstract.

In the complex tumor microenvironment, cancer stem cells (CSCs), a rare population of cells, are responsible for malignant tumor initiation, metastasis, drug resistance and recurrence. Controlling breast CSCs (BCSCs) using natural compounds is a novel potential therapeutic strategy for clinical cancer treatment. In this study, a mammosphere assay-guided isolation protocol including silica gel, a C18 column, gel filtration, and high-pressure liquid chromatography was used to isolate an inhibitory compound from Cynanchum auriculatum extracts. The isolated inhibitory compound was identified as caudatin. Caudatin inhibited breast cancer cell proliferation, mammosphere formation and tumor growth. Caudatin decreased the CD44 + /CD24 - and aldehyde dehydrogenase + cell proportions and the levels of c-Myc, Oct4, Sox2, and CD44. Caudatin induced ubiquitin (Ub)-dependent glucocorticoid receptor (GR) degradation and blocked subsequent Yes-associated protein (YAP) nuclear accumulation and target gene transcription signals in BCSCs. These results show that the GR/YAP signaling pathway regulates BCSC formation and that caudatin may be a potential chemopreventive agent that targets breast cancer cells and CSCs.

Eight new compounds (Entanutilins O-V; 1-8), including four limonoids, two steroids, one triterpenoid and one lignan were isolated from the stem barks of Entandrophragma utile. Their structures were determined by detailed spectroscopic analyses (HRESIMS and 1D/2D-NMR). Bioactivity screening indicated that compounds 1, 6 and 7 exhibited effective in reversing resistance in MCF-7/DOX cells at a nontoxic concentration of 30 μM with 18.18-, 7.43- and 7.94-fold enhancing effect, respectively, meanwhile, compounds 5 and 6 showed moderate NO inhibitory activities in LPS-activated RAW 264.7 macrophages., Competing Interests: Declaration of Competing Interest We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled., (Copyright © 2020 Elsevier B.V. All rights reserved.)

A new 9,11-secosteroid, (22R)-22-acetoxy-3β,6α,11-trihydroxy-9,11-seco-5α-cholest-7-en-9-one, along with twelve known analogues were isolated from the South China Sea gorgonian coral Iciligorgia sp. Their structures were determined by the spectroscopic analysis and comparison with reported data. In an in vitro test of osteoclastic differentiation, three compounds exhibited significant down-regulating effect whereas two compounds showed significant up-regulating effect at 0.5 and 1.0 μm. This is the first report of the chemical investigation of the gorgonian Iciligorgia sp. The acetoxy substitution at C-22 seems to play an important role in the regulating activity., (© 2020 Wiley-VHCA AG, Zurich, Switzerland.)

Two new steroids, (22R,23S)-3β-hydroxy-23-methyl-17,20-epoxyergost-5-en-22-yl acetate and (22R,23S)-5-hydroperoxy-23-methyl-5α-17,20-epoxyergost-6-ene-3β,22-diol, were isolated from the South China Sea soft coral Lobophytum sp., together with two related known ones. The structures of all compounds were elucidated by extensive spectroscopic analysis and by comparing their spectral data with those previously reported. The structure of (22R,23S)-3β-hydroxy-23-methyl-17,20-epoxyergost-5-en-22-yl acetate was further confirmed through chemical correlation. All the isolates were evaluated for the in vitro inhibitory activity against NF-κB, a potential target for the treatment of cancer, and (22R,23S)-5-hydroperoxy-23-methyl-5α-17,20-epoxyergost-6-ene-3β,22-diol exhibited moderate inhibition activity with IC 50 value of 8.96 μg/mL., (© 2020 Wiley-VHCA AG, Zurich, Switzerland.)

Six new compounds, including two new isochromane lactones, versicoisochromanes A and B (1 and 2), two new benzolactones, versicobenzos A and B (3 and 4), one furancarboxylic derivate, asperfuran A (6) and one ergosterol-type steroid, asperergoster A (7), along with five known steroids (8-12), were isolated from the Anoectochilus roxburghii endophytic fungus Aspergillus versicolor. The structures of these new compounds were determined by extensive spectroscopic techniques and electronic circular dichroism (ECD) calculations. It is notable that the new compound 7 exhibited obvious IL-1β, NO and TNF-α inhibitory activity in LPS-stimulated RAW264.7 macrophages, with IC 50 values of 35.5, 33.9 and 31.3 μM, respectively. Furthermore, compounds 7 and 8 displayed potential inhibitory effects on murine splenocytes proliferation stimulated by anti-CD3/anti-CD28 monoclonal antibodies (mAbs), meanwhile suppress the lipopolysaccharide (LPS) irritated murine splenocytes proliferation., (Copyright © 2020. Published by Elsevier B.V.)

Six undescribed steroids were isolated from the fungus Inonotus obliquus. Notably, compounds 1 and 2 represented the first example of 8,14-seco-4-methylpregnane. By spectroscopic data analyses, quantum chemical calculations and DP4+ probability analysis, their structures were unambiguously determined. The absolute configurations of the compounds 1-6 were defined by comparison of their experimental and calculated electronic circular dichroism (ECD) spectra data. The structure of compound 1 was also confirmed by single-crystal X-ray diffraction. All isolated steroids were evaluated for their neuroprotective activities against H2O2-induced cell injury in human neuroblastoma SH-SY5Y cells, among which compound 2 showed moderate activity at 12.5 μM.

Three new oxygenated steroids, sinulasterols A-C (1-3), along with seven known related steroids 4-10, were isolated from the Chinese soft coral Sinularia depressa. The structures of the new compounds were established from extensive spectroscopic data analyses and by comparison of their spectral data with those reported in the literature. Among the new compounds, metabolites 1 and 2 featured on unusual C-18 oxygenated pattern. In the TNF-α bioassay, compound 4 exhibited a potent inhibitory activity (IC 50  = 12.1 μM), which was analogous to the positive control dexamethasone (IC 50  = 8.7 μM), metabolites 1 and 2 displayed a moderate inhibitory activity (IC 50 51.1 μM and 22.7 μM respectively)., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Timosaponin B-II (TB-II; (25S)-26-(β-D-glucopyranosyloxy)-3β-[(2-O-β-D-glucopyranosyl-β-D-galactopyranosyl) oxy]-5β-furostan-22-ol is extracted from Anemarrhena . Its anti-inflammation, anti-oxidation, and anti-asthma properties have been widely explored. However, its effect on the heart has not been reported. In this study, we used zebrafish as a research model to determine the effects of TB-II on the heart and its toxic and anti-inflammatory effects. To explore the cause of cardioprotective effects of TB-II, we used transgenic zebrafish with macrophages and neutrophils labeled with fluorescent protein. We found for the first time that TB-II had a protective effect on the zebrafish heart. It did not affect the survival and hatching rates of zebrafish embryos, indicating its low toxicity. Results showed that TB-II may have cardioprotective effects, which might be related to its anti-inflammatory effects.

Penicildiones A-D (1-4), four new steroids derivatives together with three known compounds including 16α-methylpregna-17α,19-dihydroxy-(9,11)-epoxy-4-ene-3,18-dione-20-acetoxy (5), stachybotrylactone B (6) and stachybotrin (7) were isolated from the soft coral-derived fungus Penicillium sp. SCSIO41201, cultured in the 1% NaCl PDB substrate. Their structures were determined through spectroscopic methods and X-ray crystallography. Biological evaluation results revealed that 6 exhibited significant cytotoxic activity against HL-60, K562, MOLT-4, ACHN, 786-O, and OS-RC-2 cell lines with IC 50 values of 5.23, 4.12, 4.31, 23.55, 7.65 and 10.81 μmol·L -1 , respectively, while other compounds showed weak or no cytotoxicity at 50 μmol·L -1 ., (Copyright © 2020 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.)

Anemarrhena asphodeloides Bunge is a traditional Chinese medicine. The timosaponin BII is one of the most abundant and widely studied active ingredients in Anemarrhena asphodeloides Bunge. Related studies have shown that timosaponin BII has potential value for development and further utilization. The protective effect of timosaponin BII on islet β cells under type 2 diabetes was investigated in the glycolipid toxic INS-1 cell model and possible biomarkers were explored by lipidomics analysis. Timosaponin BII was isolated from Anemarrhena asphodeloides Bunge by polyamide resin and Sephadex LH-20. Then, the glycolipid toxicity INS-1 cell model was established to investigate the protective effect of timosaponin BII. The results showed that timosaponin BII could significantly influence the levels of malondialdehyde (MDA) and glutathione (GSH), thereby restoring the insulin secretion ability and cell viability of model cells. Lipidomics analysis was combined with multivariate statistical analysis for marker selection. The four most common pathological and pharmacological lipid markers were phosphatidylserine (PS), suggesting that timosaponin BII had protective effects on model cells related to the reduction oxidative stress and macrophage inflammation. RAW264.7 macrophages were stimulated by LPS to establish a model of inflammation and study the effect of timosaponin BII on the nodes of NOD-like receptor P3 (NLRP3) inflammasome pathway in the model cells. In conclusion, timosaponin BII may have the effect of protecting INS-1 pancreatic β cells through reducing IL-1β (interleukin-1β) production by inhibiting the NLRP3 inflammasome in macrophage and restoring the insulin secretion ability and cell viability by reducing oxidative stress., (© 2020 Wiley-VHCA AG, Zurich, Switzerland.)

Two new ergostane-type steroids named tiamenones A and B (1-2) were isolated from the bark of Entandrophragma angolense (Meliaceae) along with ten known compounds identified as 20S-hydroxy-4,6,24(28)-ergostatrien-3-one (3), 3β,7α,20β-trihydroxyergosta-5,24(28)-diene (4), 3β,5α-dihydroxyergosta-7,22-diene (5), stigmasterol, β-sitosterol, β-amyrin, oleanolic acid, asperphenamate, sucrose and daucosterol. The structures of the isolated compounds have been established using NMR spectroscopic and mass spectrometric analyses. The assignment of relative and absolute configurations of compound 1 was achieved by a NOESY experiment and comparison of its NMR data with those of known compound reported in literature. Compounds 1-3, β-amyrin and asperphenamate were evaluated for their antibacterial potencies against five bacterial model strains viz. Escherichia coli DSMZ 1058, Pseudomonas agarici DSMZ 11810, Bacillus subtilis DSMZ 704, Micrococcus luteus DMSZ 1605 and Staphylococcus warneri DSMZ 20036 and their cytotoxicity on two cell lines KB3-1 and HT-29. No potencies were exhibited by the tested compounds even at the highest concentration of 0.5 mg/mL. Compounds 1-3 were found to be potential HIV-1 inhibitors based on their molecular docking analyses., (Copyright © 2020 Elsevier Inc. All rights reserved.)