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Miroestrol is a chromene with potent estrogenic activity present in Pueraria candollei , commonly known as White Kwao Krua. Although this compound is only present in low amounts in the plant, it plays an important role in the estrogenic action of P. candollei products. As a tool for further studies about the efficacy and safety of P. candollei as a phytoestrogenic supplement, we generated a novel monoclonal antibody against miroestrol. This anti-miroestrol monoclonal antibody was used to develop an immunoassay for the determination of miroestrol content, which can be used for quality control purposes of P. candollei . The developed ELISA against miroestrol has a calibration range of 10-780 ng/mL miroestrol, a limit of detection of 3.5 ng/mL, and a limit of quantitation of 12.2 ng/mL. According to the validation analysis, the established ELISA is precise, accurate, specific, and sensitive for miroestrol detection in plants. Furthermore, the anti-miroestrol monoclonal antibody was used to prepare an immunoaffinity column for the isolation of miroestrol from the tuberous root of P. candollei . The column provides a simple procedure for miroestrol isolation, with a capacity of 3.91 µg of miroestrol per 1 mL of immunogel., Competing Interests: Conflict of Interest: The authors declare no conflict of interest., (Georg Thieme Verlag KG Stuttgart · New York.)

Bulbus of Fritillaria pallidiflora, the dried bulb of F. pallidiflora, is widely used in Chinese folk medicine due to its powerful biological activities. The aim of this study was to investigate in vitro and in vivo antitumor activity of different fractions and isosteroidal alkaloids from bulbus of F. pallidiflora and clarify its putative mechanism of antitumor activity. Firstly, we assayed in vitro antitumor effects of different fractions from bulbus of F. pallidiflora and found that chloroform extracts and purified total alkaloids of bulbus of F. pallidiflora showed higher cytotoxic activity than other tested extracts. We further isolated four main alkaloids, chuanbeinone, imperialine-β-N-oxide, isoverticine and isoverticine-β-N-oxide, from the total alkaloids of bulbus of F. pallidiflora and found that they display significant cytotoxicity, whereby chuanbeinone showed the highest activity against Lewis lung carcinoma cells. Moreover, we found that chuanbeinone induced S phase arrest and further increased apoptosis of Lewis lung carcinoma cells. The results of Western blotting experiments showed that the expression of the antiapoptotic Bcl-2 was reduced by chuanbeinone treatment, while the proapoptotic protein Bax and caspase-3 were increased. Moreover, we investigated the in vivo antitumor activity of chuanbeinone and characterized its putative antitumor mechanism of action by the TUNEL assay and by histological and immunohistochemical analyses. Our results showed that chuanbeinone exhibited significant antitumor activity in vivo, while notably inhibiting tumor angiogenesis and inducing apoptosis characterized by an increased expression of caspase-3. Our findings show that chuanbeinone exhibits significant antitumor activity in vitro and in vivo, and points to possible therapeutic potential for this compound as well as for its natural source, bulbus of F. pallidiflora., (Georg Thieme Verlag KG Stuttgart · New York.)

Six new taccalonolides, taccalonolides AT-AY (1-6), and two new withanolides, chantriolides D and E (7 and 8), together with ten known compounds (9-18), have been isolated from whole plants of Tacca chantrieri. The structures, including the absolute configurations of some of the compounds, were determined by spectroscopic and chemical methods. All compounds were evaluated for their in vitro cytotoxicity against five tumor cell lines. Compounds 9, 10, 13-15, and 17 exhibited cytotoxic activity, with IC50 values of 1.13-5.71 µM, while compound 7 showed selective cytotoxicity. The results indicated that taccalonolides with a six-membered lactone moiety located at C-15 and C-24 were devoid of cytotoxicity against five tumor cell lines (> 10 µM)., (Georg Thieme Verlag KG Stuttgart · New York.)

Five new compounds (1-5), including two limonoids, one triterpenoid, one steroid, and one sesquiterpenoid, along with nine known limonoids (6-14), were isolated from the bark of Melia azedarach. The structures of the new compounds were elucidated by 2D NMR spectroscopy and mass spectrometry. The isolated compounds as well as three acetylated derivatives of 9 were evaluated for their cytotoxicities against five human tumor cell lines (HL-60, SMMC-7721, A-549, MCF-7, and SW480) by an MTT assay. Seven limonoids (1, 6, 7, 8, 9, 9b, and 9c) showed significant inhibitory activities against tested cell lines with IC50 values ranging from 0.003 to 0.555 µM, and their preliminary structure-activity relationships are also discussed., (Georg Thieme Verlag KG Stuttgart · New York.)

Five new steroidal glycosides, timosaponin J ( 1), timosaponin K ( 2), (25 S)-karatavioside C ( 5), timosaponin L ( 6), and (25 S)-officinalisnin-I ( 8), together with eight known steroidal saponins, timosaponin E (1) ( 3), purpureagitosid ( 4), timosaponin BII ( 7), timosaponin B III ( 9), anemarrhenasaponin I ( 10), anemarrhenasaponin III ( 11), anemarrhenasaponin A (2) ( 12), and timosaponin A III ( 13), were isolated from the rhizomes of Anemarrhena asphodeloides. Their structures were elucidated on the basis of spectroscopic and chemical evidence. The aglycones of compounds 1 and 2 are new aglycones. Compounds 1- 13 were evaluated for their platelet aggregation activities, and compound 13 exhibited the strongest inhibitory effect on adenosine diphosphate (ADP)-induced platelet aggregation., (© Georg Thieme Verlag KG Stuttgart · New York.)

Two new steroidal saponins, 8 and 10, along with 7 known steroidal sapogenins and saponins (1-7) and a furostanol saponin (9) were isolated from Agave sisalana Perrine ex Engelm. The structures of these two new compounds were identified and characterized by 1D and 2D NMR spectroscopy and mass spectrometry. In addition, acid hydrolysis and GC-FID were used to confirm the sugar moieties of 8 and 10. The cytotoxic effects of 1-10 on MCF-7, NCI-H460, and SF-268 cancer cells were evaluated, and among them, compound 10 proved to be the most cytotoxic with IC₅₀ values of 1.2, 3.8, and 1.5 µM, respectively., (© Georg Thieme Verlag KG Stuttgart · New York.)

Two new triterpenoids (1, 2) and two new steroids (3, 4) along with twelve related known compounds (5-16) were isolated from the bark of Melia azedarach. The new structures were elucidated by means of spectroscopic methods and molecular modeling studies and found to be 21,24-cycloeupha-7-ene-3 β,16 β,21 α,25-tetrol (1), 3 β-acetoxy-12 β-hydroxy-eupha-7,24-dien-21,16 β-olide (2), 29-hydroperoxy-stigmasta-7,24(28) E-dien-3 β-ol (3), and 24 ξ-hydroperoxy-24-vinyl-lathosterol (4). All isolated compounds were tested for their cytotoxic activity against three human cancer cell lines (A549, H460, HGC27) using the CellTiter Glo™ luminescent cell viability assay. Among them, compounds 2- 4, 24 ξ-hydroperoxy-24-vinyl-cholesterol (6), kulinone (7), meliastatin 3 ( 8), 3-oxo-olean-12-en-28-oic acid (10), and (22 E,24 S)-5 α,8 α-epidioxy-24-methyl-cholesta-6,22-dien-3 β-ol (12) were found to have cytotoxic effects, with IC₅₀ values of 5.6-21.2 µg/mL., (© Georg Thieme Verlag KG Stuttgart · New York.)

The steroids ergone (1), (22E, 24R)-ergosta-7,22-dien-3β-ol (2), 5α,8α-epidioxy-(22E,24R) -ergosta-6,22-dien-3β-ol (3), ergosta-6,22-dien-3β,5α,6β-triol (4), and polyporusterone B (5) were isolated from Polyporus umbellatus by bioassay-guided approach. They showed potent anticancer activity against HepG2 cells. Ergone displayed remarkable anticancer activity against HepG2, Hep-2, and Hela cancer cells, of which HepG2 cells were the most sensitive. Furthermore, the cytotoxic effects of ergone on normal human cells (HUVEC) were smaller than on cancer cells. The results showed that ergone had more selective cytotoxic activity against cancer cells than against normal cells., (© Georg Thieme Verlag KG Stuttgart · New York.)

The 95 % ethanol extract of Gelsemium sempervirens showed inhibitory activity against human DNA topoisomerase I (Topo I). Phytochemical investigations of this active extract resulted in the isolation and identification of three new steroids ( 1 - 3), together with eight known compounds 12 beta-hydroxy-5 alpha-pregn-16-ene-3,20-dione ( 4), gelsemine ( 5), sempervirine ( 6), scopoletin ( 7), 7- O- beta- D-glucopyranosylscopoletin ( 8), 7- O- beta- D-apiofuranosyl-(1-->6)- beta- D-glucopyranosylscopoletin ( 9), uvaol ( 10), and 2-(4-hydroxyphenyl)ethyl heptadecanoate ( 11). The structures of the new steroids were determined by extensive NMR and HR-ESI-MS analyses as 21-hydroxy-5 alpha-pregn-16-ene-3,20-dione ( 1), 3-oxoandrosta-16-ene-17-carboxylic acid ( 2), and 3-oxoandrosta-4,16-diene-17-carboxylic acid ( 3). This study suggests that sempervirine ( 6) intercalates to DNA and also inhibits Topo I through modulating the enzyme activity with an IC (50) of 54.5 +/- 15.9 muM.

In addition to the known sterols and ketosteroids beta-sitosterol (24 alpha-ethylcholest-5-en-3 beta-ol), stigmasterol (24 alpha-ethylcholesta-5,22-dien-3 beta-ol), campesterol (24 alpha-methylcholest-5-en-3 beta-ol), beta-sitostenone (stigmast-4-en-3-one, 24 alpha-ethylcholest-4-en-3-one), stigmastenone (stigmasta-4,22-dien-3-one, 24 alpha-ethylcholesta-4,22-dien-3-one), campestenone (24 alpha-methylcholest-4-en-3-one), and stigmasta-3,5-dien-7-one (24 alpha-ethylcholesta-3,5-dien-7-one), the new steroids stigmasta-3,5,22-trien-7-one (24 alpha-ethylcholesta-3,5,22-trien-7-one), and campesta-3,5-dien-7-one (24 alpha-methylcholesta-3,5-dien-7-one) were isolated from the stem bark of Harrisonia abyssinica and identified by NMR and mass spectrometry.

By bioactivity-guided fractionation, three known steroidal glycosides, beta 2-solamargine (1), solamargine (2), and degalactotigonin (3), were isolated from Solanum nigrum. Compounds 1-3 induced morphological abnormality in Pyricularia oryzae mycelia with MMDC (minimum morphological deformation concentration) values of 63.0, 38.5, and 97.2 microM, respectively. This is the first report on the isolation of 1 from this species and on the cytotoxicity of 1-3 on six cultured human solid tumor cell lines HT-29 (colon), HCT-15 (colon), LNCaP (prostate), PC-3 (prostate), T47D (breast), and MDA-MB-231 (breast) in vitro. The cytotoxic assay indicated that 2 is the main antineoplastic agent in S. nigrum. The structures of 1-3 were elucidated on the basis of chemical evidence and spectral analysis, especially by 2D-NMR analysis.

Two oligofurostanosides were isolated from the seeds of Asparagus officinalis L and their structures characterized as 3-O-[alpha-L-rhamnopyranosyl-(1-->2)-(alpha-L-rhamnopyranosyl- (1-->4))-beta-D-glucopyranosyl]-26-O-[beta-D-glucopyranosyl]-(25R) -22 alpha-methoxyfurost-5-ene-3 beta,26-diol(methyl protodioscin) and its corresponding 22 alpha-hydroxy analogue (protodioscin). The structural identification was performed using detailed analysis of 1H- and 13C-NMR spectra including two-dimensional NMR spectroscopy (COSY, HMQC, NOESY and HMBC), and chemical conversions. These two compounds have been shown to inhibit the growth of human leukemia HL-60 cells in culture and macromolecular synthesis in a dose-dependent manner. The inhibitory effect on DNA synthesis was found to be irreversible.

Twenty-nine C-27 steroidal saponins were tested for the inhibitory effects on human spermatozoa in vitro by means of a modified Sander-Cramer method. Twelve of them are responsible for this activity. The structure-activity relationship is discussed.

The antinociceptive actions of the steroid compounds isolated from the leaves, stems, and roots of P. corcovadensis have been investigated in mice. Stigmasterol, stigmasterol acetate, beta-sitosterol, and aspirin (3-100 mk/kg, i.p.) inhibited, in a dose-related manner, acetic acid-induced abdominal constriction in mice with ID50s of 16, 11, 9, and 24 mg/kg, respectively. In the formalin test, stigmasterol and stigmasterol acetate (10-100 mg/kg, i.p.) caused graded inhibition of both the neurogenic (first phase) and inflammatory phases (second phase) of formalin-induced pain. However, both compounds were more effective in relation of the second phase of the formalin test with ID50 values of 26 and 41 mg/kg, respectively. Furthermore, both steroids failed to affect the edematogenic response of the formalin test. Given orally, stigmasterol and stigmasterol acetate (50-200 mg/kg) also exhibited significant though less potent analgesic action against both acetic acid- and formalin-induced nociception in mice. In addition, stigmasterol (up to 100 mg/kg, i.p.), in contrast to morphine (10 mg/kg, s.c.), had no analgesic effect in either tail-flick or hot-plate models. These findings suggest that stigmasterol and beta-sitosterol may account, at least in part, for the antinociceptive actions reported previously for the hydroalcoholic extract of Phyllanthus corcovadensis.