Anemarrhenasaponin I

Steroidal glycosides from the rhizomes of Anemarrhena asphodeloides and their antiplatelet aggregation activity

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.

Study on the Effects of Chinese Materia Medica Processing on the Hypoglycemic Activity and Chemical Composition of Anemarrhenae Rhizoma

Purpose: To compare the hypoglycemic effects of different extracts of Anemarrhenae Rhizoma (AR) before and after being stir-baked with salt water on the diabetic mice and to detect the contents of 8 components in the corresponding active parts simultaneously using the UPLC-MS method, in order to screen the better extracts for diabetes and to clear the material basis for enhancing hypoglycemic activity of Anemarrhenae Rhizoma stir-baked with salt water (SAR).
Methods: Taking spontaneous type II diabetic db/db mice as models and fasting blood glucose (FBG), oral glucose tolerance test (OGTT), glycated hemoglobin or glycosylated hemoglobin (HbAlc), serum resistin (RESISTEIN), fasting insulin (FINS), superoxide dismutase (SOD), malondialdehyde (MDA), and nitric oxide (NO) as indicators, the hypoglycemic effects of different active parts of Anemarrhenae Rhizoma were evaluated. The chromatographic separation was performed on a Waters BEH C18 (2.1 mm × 50 mm, 1.7 μm) column using acetonitrile (B) and 0.1% formic acid in water (A) as mobile phases, and the flow rate was 0.3 ml/min. The column temperature was set as 28°C, and the injection volume was 10 μL. A mass spectrometer was connected to the UPLC system via an electrospray ionization (ESI) interface. Full-scan data acquisition was performed in the negative ion mode.
Result: In the study of pharmacodynamics, the hypoglycemic effect of Anemarrhenae Rhizoma stir-baked with salt water is better than that of Anemarrhenae Rhizoma and the hypoglycemic effect of ethanol extract of Anemarrhenae Rhizoma is more remarkable than that of the decoction. The measured components all have a good linear relationship within their respective linear ranges (r ≥ 0.9990); the average recovery rates are 98.86%-100.69%, RSD <2.90%. Compared with the raw Anemarrhenae Rhizoma, the contents of Timosaponin AIII, Timosaponin BII, Timosaponin BIII, Anemarrhenasaponin I, Anemarrhenasaponin Ia, and Mangiferin of Anemarrhenae Rhizoma stir-baked with salt water are all higher, the changes of Timosaponin AI and Anemarrhenasaponin AII are not obvious, and all the contents of chemical composition in the ethanol extract of Anemarrhenae Rhizoma and Anemarrhenae Rhizoma stir-baked with salt water were obviously higher compared with the water decoction.
Conclusion: The processing method, stir-baking with salt water, can increase the contents of active compositions in Anemarrhenae Rhizoma and strengthen the hypoglycemic effect. The ethanol extract of Anemarrhenae Rhizoma stir-baked with salt water is the better active site.

Effect of six steroidal saponins isolated from anemarrhenae rhizoma on platelet aggregation and hemolysis in human blood

Six steroidal saponins were isolated from Anemarrhena asphodeloides Bunge (Liliaceae), a traditional chinese medicine, and named anemarrhenasaponin I (An-I), anemarrhenasaponin Ia (An-Ia), timosaponin B-I (TB-I), timosaponin B-II (TB-II), timosaponin B-III (TB-III), and timosaponin A-III (TA-III). The effects of these six compounds on platelet aggregation and hemolysis in human blood were studied. All these compounds provoked remarkable inhibiting effect on platelet aggregation, and activated partial thromboplastin times (APTT) are sensitive to the presence of these six compounds. Using an in vitro system, APTT was delayed with the increment of the concentrations of these six compounds. In these six compounds, only timosaponin A-III appeared a strong effect on hemolysis, and anemarrhenasaponin Ia had a slight effect on hemolysis, other had no effect on hemolysis. These results suggested that these steroidal saponins isolated from Anemarrhena asphodeloides Bunge (Liliaceae) might be used as a novel antithrombotic therapeutic agents in post-myocardial infarction.

Effect of steroidal saponins of Anemarrhenae rhizoma on superoxide generation in human neutrophils

Effect of six steroidal saponins isolated from Anemarrhenae rhizoma on superoxide generation in human neutrophils was investigated. The steroidal saponins examined were anemarrhenasaponin-I (An-I), anemarrhenasaponin-Ia (An-Ia), timosaponin B-I (TB-I), timosaponin B-II (TB-II), timosaponin B-III (TB-III) and timosaponin A-III (TA-III). An-I, An-Ia, and TB-III suppressed the superoxide generations induced by N-formyl-methionyl-leucyl-phenylalanine (fMLP) and arachidonic acid (AA) in a concentration-dependent manner, but enhanced that induced by phorbol 12-myristate 13-acetate (PMA). While TB-II also suppressed and enhanced the superoxide generations induced by fMLP and PMA, respectively, the compound significantly enhanced the AA-induced superoxide generation. TB-I enhanced the fMLP-induced superoxide generation in a low concentration range (peak at 40 microM), gave no effect on the PMA-induced superoxide generation and weakly enhanced the AA-induced superoxide generation. TA-III enhanced the fMLP-induced superoxide generation more than twice as much as that by TB-I in the same concentration range. However, TA-III enhanced the PMA-induced superoxide generation and most significantly suppressed the AA-induced superoxide generation.

Characterization of spirostanol glycosides and furostanol glycosides from anemarrhenae rhizoma as dual targeted inhibitors of 5-lipoxygenase and Cyclooxygenase-2 by employing a combination of affinity ultrafiltration and HPLC/MS

Background: Modulation of the arachidonic acid (AA) cascade via 5-lipoxygenase (5-LOX) and cyclooxygenase-2 (COX-2) represent the two major pathways for treatments of inflammation and pain. The design and development of inhibitors targeting both 5-LOX and COX-2 has gained increasing popularity. As evidenced, 5-LOX and COX-2 dual targeted inhibitors have recently emerged as the front runners of anti-inflammatory drugs with improved efficacy and reduced side effects. Natural products represent a rich resource for the discovery of dual targeted 5-LOX and COX-2 inhibitors. By combining affinity ultrafiltration and high-performance liquid chromatography-mass spectrometry (AUF-LC-MS), an efficient method was developed to identify spirostanol glycosides and furostanol glycosides as the 5-LOX/COX-2 dual inhibitors from saponins extract of Anemarrhenae Rhizoma (SEAR).
Methods: A highly efficient method by combining affinity ultrafiltration and high-performance liquid chromatography-mass spectrometry (AUF-LC-MS) was first developed to screen and characterize the 5-LOX/COX-2 dual targeted inhibitors from SEAR. The structures of compounds in the ultrafiltrate were characterized by high resolution Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS). In addition, in vitro 5-LOX/COX-2 inhibition assays and their dual expression in vivo were performed to confirm the inhibitory activities of the compounds screened by AUF-LC-MS. Molecular docking studies with the corresponding binding energy were obtained which fit nicely to both 5-LOX and COX-2 protein cavities and in agreement with our affinity studies.
Results: A total of 5 compounds, timosaponin A-II, timosaponin A-III, timosaponin B-II, timosaponin B-III and anemarrhenasaponin I, were identified as potential 5-LOX/COX-2 dual targeted inhibitors with specific binding values > 1.5 and IC₅₀ ≤ 6.07 μM.
Conclusion: The present work demonstrated that spirostanol glycoside and furostanol glycoside were identified as two novel classes of dual inhibitors of 5-LOX/COX-2 enzymes by employing a highly efficient screening method of AUF-LC-MS. These natural products represent a novel class of anti-inflammatory agents with the potential of improved efficacy and reduced side effects.