The genus Rabdosia is famous for the abundance of diverse and novel ent-kaurane diterpenoids. However, only a few ent-kauranoids have been discovered from R. flexicaulis since the investigation on its chemical constituents is not systematic. To find novel bioactive diterpenoids, the ethyl acetate extract of the above ground part of R. flexicaulis in Daofu County, Sichuan Province was obtained by column chromatography. One new compound and five known ones were identified as flexicaulin E(1), forrestin B(2), inf-lexarabdonin D(3), 7α-hydroxydehydroabietic acid(4), 15-hydroxydehydroabietic acid(5), and pomiferin F(6) by spectral techniques. Compounds 1-3 were the ent-kaurane diterpenoids isolated from this species for the first time. Compounds 4-6, aromatic abie-tanoids, were isolated from the genus Rabdosia for the first time.
Diterpenes ; Diterpenes, Kaurane ; Isodon/chemistry* ; Molecular Structure ; Plant Extracts/chemistry*

OBJECTIVETo prepare the oridonin submicron emulsion and characterize their properties.

METHODHigh pressure homogenization method was employed to prepare the oridonin submicron emulsion and such properties as size, Zeta potential and viscosity were characterized.

RESULTThe results showed that the submicron emulsions was formed with the drug loading 1 g L(-1), particle size of (138.87 +/- 0.60) nm, zeta potential of (47.27 +/- 2.31) mV, pH value of (6.02 +/- 0.03) and viscosity of (1.78 +/- 0.015) MPa s, respectively.

CONCLUSIONThe method is feasible and the submicron emulsions has stable properties. Experiments offer a new formulation of oridonin for clinical application.

Chemistry, Pharmaceutical ; Diterpenes, Kaurane ; chemistry ; Emulsions ; chemistry ; Particle Size ; Viscosity

To study the chemical constituents of Fritillaria monanth Migo, the constituents were separated and purified by column chromatography on silica gel, and the structures were identified by NMR, MS spectral data. Six compounds were isolated and identified as ent-kauran-15-en-17-ol (I), entkauran-15-en-3alpha, 17-diol (II), fritillaziebinol (III), ent-kauran-16a, 17-diol (IV), ent-kauran-3alpha, 16alpha,17-triol (V), ent-16,17-epoxy-kauran-3alpha-ol (VI). All the compounds were isolated from this plant for the first time, and VI is named as ent-16,17-epoxy-kauran-3alpha-ol, which is a new compound.
Diterpenes, Kaurane ; chemistry ; isolation & purification ; Fritillaria ; chemistry ; Molecular Structure ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Diterpenes, Kaurane ; pharmacology ; Humans ; Nasopharyngeal Neoplasms ; pathology

OBJECTIVETo develop a HPLC method for the determination of plasma concentration of oridonin (ORI) and study the pharmacokinetics of ORI in mice.

METHODBlood was sampled from mice which were injected ORI by 10 mg x kg(-1) at different time intervals, and the concentration of ORI was determined by HPLC. The pharmacokinetic parameters were accessed by 3P97.

RESULTThe calibration curve was linear (r = 0.998 7) within the range of 0.202-20.0 mg x L(-1) for ORI in plasma. The average recoveries were more than 93%. The within-day and between-day precisions were no more than 9%. After i.v. oridonin in mice, the plasma concentration-time course fitted well to two-compartment model. The pharmacokinetic equation was C = 16.192 5e(-0.554 6t) + 5.475 7e(-0.016 3t). The pharmacokinetic parameters were below: t1/2alpha 1.249 9 min, t1/2beta 42.638 4 min, K21 0.152 3 min(-1), K12 0.359 3 min(-1), K10 0.0592 min(-1), AUC 366.035 0 microg x min x mL(-1), CL 0.0273 L x min(-1) x kg(-1), V(c)0.461 5 L x kg(-1).

CONCLUSIONThe method can be used to determine the concentration and to investigate the pharmacokinetics of ORI in mice. ORI was absorbed and distributed very fast in mice. The effect of ORI was rapid. The elimination was the main process.

Animals ; Chromatography, High Pressure Liquid ; methods ; Diterpenes, Kaurane ; pharmacokinetics ; Female ; Male ; Mice

OBJECTIVETo investigate the stability of oridonin (ORI) solution for research and development of novel ORI prepartions.

METHODThe effect of pH on the degradation rate of ORI was evaluated, the pH values of the oridonin solutions were adjusted to the setting pH, with 1 mol x L(-1) HCl or NaOH, respectively, and stored at room temperature for 60 h. The constant temperature method was applied to evaluate the stability of ORI solution at room temperature and at 4 degrees C.

RESULTThe pH-rate profile of ORI was V-shaped, and the pHm was 5. The t90 of ORI solution at room temperature was 53.2 h and 91.5 h at 4 degrees C CONCLUSION: The ORI solution is not stable. The pH-dependent degradation of ORI solution confirms to specific acid-base catalysis reaction.

Diterpenes, Kaurane ; chemistry ; Drug Stability ; Hydrogen-Ion Concentration ; Solubility ; Solutions ; Temperature ; Time Factors ; Water ; chemistry

OBJECTIVETo investigate the release characteristics and mechanism of oridnonin self-microemulsifying drug delivery system (SMEDDS) in vitro.

METHODThe concentration of oridonin was determined by HPLC. In vitro release studies were conducted by reverse dialysis technique. The effects of release medium, agitation rate and preparations on the oridonin release were studied. The similarity factor (f2) was applied to the release profile comparisons. Model fitting was used to determine the kinetics and mechanism.

RESULTThe release media and agitation rate from 50-100 r x min(-1) had no distinctive effect on the oridonin release kinetics, which the similarity factors were greater than 50. The oridonin release profiles for oridonin SMEDDS and oridonin ethanol solution were similar. 65% of oridonin were released in 30 min for oridonin SMEDDS in pH 7.8 PBS. Oridonin SMEDDS fit the Hixson-Crowell model best.

CONCLUSIONThe release data from oridonin SMEDDS showed it release fast. The deduced release mechanism is that the surface and particle sizes of self-microemulsion in water solution are changing during the process of release and the drug penetration through membrane is a passive diffusion process.

Chromatography, High Pressure Liquid ; Diterpenes, Kaurane ; chemistry ; Drug Delivery Systems ; methods ; Emulsions ; chemistry ; Kinetics

OBJECTIVETo study the chemical constituents of Tibetan medicine Caryopteris toroetii.

METHODThe crude drug was extracted with 95% EtOH and isolated by repeated chromatographic methods. The structures of the isolated compounds were elucidated on the basis of spectral analysis.

RESULTTwo en-kaurene diterpenoid compounds: oridonin (1) and nodosin (2), were obtained from C. toroetii and their 1H and 13C-NMR data in CD3OD were reported for the first time.

CONCLUSIONThe en-kaurene diterpenoid compounds were obtained from genus Caryopteris for the first time.

Diterpenes, Kaurane ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Lamiaceae ; chemistry ; Medicine, Tibetan Traditional

OBJECTIVE: To investigate the effects of oridonin (ORI) on the proliferation and apoptosis of human multiple myeloma cell line H929 and its possible mechanism. METHODS: H929 cells were exposed to ORI 0、4、8、12、16、20、24、28、32 μmol/L for 12, 24 and 36 hours respectively. The prolifcration inhibitory effect of ORI on H929 cells was determined by MTT assay and then the working concentrations of ORI were determined. The morphological changes and apoptosis of H929 cells were observed by TUNEL (TdT-mediated dUTP Nick-End Labeling) and fluorescence microscopy. The apoptosis rate of H929 cells was detected by flow cytometry with Annexin V-FITC/PI staining. The protein expressions of pro-caspase-3, BCL-2，p-PI3K, p-Akt, BAX, Cleaved PARP and p-JNK, p-ERK and p-p38 in H929 cells were detected by Western blot. RESULTS: Compared with the control group, the proliferation of H929 cells treated with the ORI of 8-16 μmol/L was significantly inhibited and the apoptosis of H929 cells was obviously increased in dose- and time-dependent manners. As for morphological changes, the characteristics of apoptotic cells were presented in H929 cells treated with ORI for 24 hours. The protein levels of pro-caspase-3, BCL-2，p-PI3K, p-Akt were down-regulated with increasing of ORI concentration（r=0.9861, r=0.9725, r=0.9413, r=0.9373）, while the BAX, Cleaved PARP and p-JNK, p-ERK and p-p38 were up-regulated（r=0.9178, r=0.8877, r=0.882, r=0.9645, r=0.8623）. CONCLUSION The ORI possesses anti-myeloma effects, can inhibit the proliferation and induce the apoptosis of H929 cell line in vitro. Its potential mechanism may be related with up-regulating the MAPK and down-regulating the PI3K/Akt signal pathways.
Apoptosis ; Caspase 3 ; Cell Line, Tumor ; Cell Proliferation ; Diterpenes, Kaurane ; Humans ; Multiple Myeloma ; Phosphatidylinositol 3-Kinases

OBJECTIVE: To investigate the influence of oridonin on the killing activity of NK-92 MI cells targeting THP1 and the related mechanism. METHODS: The killing activity of NK-92 MI to THP1 before and after oridonin treatment was detected by LDH release assay; the expression of natural killer cell ligands activating receptor D (NKG2D, including MICA, MICB, ULBP1, ULBP2 and ULBP3) was detected by real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot respectively; the expression of cytokine TNF-α, TNF-β and IFN-γ in the co-culture supernatant of NK-92 MI cells and THP1 cells were measured by ELISA. RESULTS: The killing efficiency after oridonin treatment at different effector-target ratio (1:1, 5:1, 10:1) was all significantly up-regulated in comparison with that before oridonin treatment (P＜0.05). QRT-PCR and Western blot showed that the expressions of mRNA and protein levels of MICB, ULBP1, ULBP2 increased to varying degree (P＜0.05), but the expression levels of MICA and ULBP3 were not statistically significant between experimental group and control group (P＞0.05). ELISA results indicated that IFN-γ and TNF-β release were significantly increased after oridonin treatment (P＜0.05), however, the TNF-α release was not statistically different in comparison with control group (P＞0.05). CONCLUSION Oridonin can significantly improve killing efficiency of NK-92 MI on THP1, that might be related with up-regulation of MICB, ULBP1 and ULBP2 expression and promotion of IFN-γ and TNF-β release.
Cell Line, Tumor ; Diterpenes, Kaurane ; pharmacology ; GPI-Linked Proteins ; Histocompatibility Antigens Class I ; Humans