Saikogenin A

Integrative strategy for quality control of Radix Bupleuri based on non-targeted metabolomic profiling and molecular networking.[Pubmed:36602568]

Anal Bioanal Chem. 2023 Feb;415(5):961-974.

Quality control of Radix Bupleuri (RB) can be challenging due to the complexity of origin, the similar morphological characteristics, and the diversity of the multiple components. In this study, an integrated strategy for extensive identification of metabolites in plants based on multiple data processing methods was proposed to distinguish four commercially available RB species. First, the pre-processed mass spectrometry data was uploaded to Global Natural Products Social Molecular Networking (GNPS) for spectral library search and molecular network analysis, which can effectively differentiate isomers and reduce molecular redundancy. Second, the possible cleavage mode was summarized from the characteristic MS/MS fragment ions of saikoside standard, and then the possible structure of saikoside in the sample was deduced according to the cleavage patterns. Third, collected all kinds of RB components reported in the literature and matched the information in the samples to obtain more comprehensive information about metabolites. Finally, chemical markers were found employing chemometrics. This strategy not only increases the variety and number of identified components, but also improves the accuracy of the data. Based on this strategy, a total of 132 components were identified from different species of RB, and 14 chemical constituents were considered to be potential chemical markers to distinguish four kinds of RB. Among them, Saikogenin A, hydroxy-saikosaponin a, hydroxy-saikosaponin d, and rutinum were of great significance for identification. The method proposed in this study not only successfully identified and distinguished four species of RB, but also laid a good theoretical foundation for regulating the RB market. This strategy provides promising perspectives in the accurate analysis of the ingredients of traditional Chinese medicine.

Clarify the potential cholestatic hepatotoxicity components from Chinese Herb Medicine and metabolism's role via hBSEP vesicles and S9/hBSEP vesicles.[Pubmed:35101544]

Toxicol In Vitro. 2022 Apr;80:105324.

In this study, the inhibitory effect of components from Chinese Herb Medicine (CHMs) with potential hepatotoxicity was assessed by human bile salt export pump (hBSEP) vesicles with and without S9 metabolism. Sixty-three compounds from 22 hepatoxicity CHMs were selected as the test articles. In hBSEP vesicles, eighteen of them were found to have moderate or strong inhibitory effect towards BSEP. Further studies were performed to determine the IC(50) values of strong inhibitors. For the compounds belong to CHMs reported to cause cholestasis and strong inhibitors defined in hBSEP vesicles, their relative transport activities of Taurocholic acid (TCA) were evaluated in hBSEP vesicles as well as hBSEP vesicles with S9 system (S9/hBSEP vesicles). The differences of their relative transport activities of TCA between the above two system were compared to reveal the net effect of metabolism on BSEP's activity. It was found that the inhibitory effect of Saikogenin A (SGA), Saikogenin D (SGD), Diosbulbin B (DB) and rhein were significantly increased; while the inhibitory effect of isobavachalcone, saikosaponin d and saikosaponin b2 were significantly decreased after S9 metabolizing. Identification of metabolic pathways suggested that CYP3A4 was responsible for aggravating inhibitory effect of SGA and SGD against BSEP.

A new triterpenoid saponin from Clinopodium chinense (Benth.) O. Kuntze.[Pubmed:26511166]

Nat Prod Res. 2016;30(9):1001-8.

A new triterpene saponin, 3beta,16beta,23alpha,28beta,30beta-pentahydroxyl-olean-11,13(18)-dien-3beta-yl-[beta-D-glucopyranosyl-(1-->2)]-[beta-D-glucopyranosyl-(1-->3)]-beta-D-fucopyranoside, was named Clinoposaponin D (1), together with six known triterpene saponins, buddlejasaponin IVb (2), buddlejasaponin IVa (3), buddlejasaponin IV (4), clinopodisides D (5), 11alpha,16beta,23,28-Tetrahydroxyolean-12-en-3beta-yl-[beta-D-glucopyranosyl-(1-->2)]-[beta-D-glucopyranosyl-(1-->3)]-beta-D-fucopyranoside (6) and proSaikogenin A (7), and two known triterpenes, Saikogenin A (8) and saikogenin F (9) were isolated from Clinopodium chinense (Benth.) O. Kuntze. Their structures were elucidated on the basis of 1D, 2D NMR and MS analysis. Meanwhile, the effects of all compounds on rabbit platelet aggregation and thrombin time (TT) were investigated in vitro. Compounds 4 and 7 had significant promoting effects on platelet aggregation with EC50 value at 53.4 and 12.2 muM, respectively. In addition, the highest concentration (200 muM) of compounds 2 and 9 shortened TT by 20.6 and 25.1%, respectively.

Cytotoxic activity and inhibitory effect on nitric oxide production of triterpene saponins from the roots of Physospermum verticillatum (Waldst & Kit) (Apiaceae).[Pubmed:19467877]

Bioorg Med Chem. 2009 Jul 1;17(13):4542-7.

Three triterpene saponins isolated from the roots of Physospermum verticillatum and identified as saikosaponin a (1), buddlejasaponin IV (2), and songarosaponin D (3) were investigated in vitro for their cytotoxic activity against a panel of seven different cancer cell lines including ACHN, C32, Caco-2, COR-L23, A375, A549, and Huh-7D12 cell lines. The hydrolysis of sugar unit was performed on saikosaponin a (1) to obtain saikosapogenin a (4). All isolated saponins exhibited strong cytotoxic activity against COR-L23 cell line with IC(50) values ranged from 0.4 to 0.6 microM. A similar activity was recorded for Saikogenin A (4). None of the tested compounds affected the proliferation of skin fibroblasts 142BR suggesting a selective action against cancer cells. Moreover, buddlejasaponin IV (2) and songarosaponin D (3) exerted significant inhibition of NO production in LPS induced RAW 264.7 macrophages with IC(50) of 4.2 and 10.4 microM, respectively.

Isolation of saponins with the inhibitory effect on nitric oxide, prostaglandin E2 and tumor necrosis factor-alpha production from Pleurospermum kamtschaticum.[Pubmed:16141537]

Biol Pharm Bull. 2005 Sep;28(9):1668-71.

As an attempt to search for bioactive natural products exerting antiinflammatory activity, we have isolated two saponins were isolated from the aerial portion of Pleurospermum kamtschaticum (Umbelliferae) by nitrite assay activity-directed chromatographic fractionation. They were identified as saikogenin F 3-O-beta-D-glucopyranosyl-(1-->2)-[beta-D-glucopyranosyl-(1-->3)]-beta-D-fucopyranoside (buddlejasaponin IV, 1) and 3beta,16beta,23,28-tetrahydroxy-11alpha-methoxyolean-12-ene 3-O-beta-D-glucopyranosyl(1-->2)-[beta-D-glucopyranosyl(1-->3)]-beta-D-fucopyranoside (buddlejasaponin IVa, 2). Compound 1 significantly inhibited nitric oxide (NO) production, and it also significantly decreased prostaglandin E2 (PGE2) and tumor necrosis factor-alpha (TNF-alpha) release in the lipopolysaccharide (LPS)-activated macrophage Raw 264.7 cells whereas compound 2 was much less active. Saikogenin A (3) and -H (4) were obtained by hydrolyzing 1 and 2. Although these sapogenin showed strong NO inhibition, these effects were caused by the cytotoxic effect on Raw 264.7 cells. These results supported the notion that buddlejasaponin IV is a major inhibitors of NO, PGE2 and TNF-alpha production in P. kamtschaticum.

Determination of saikosaponin derivatives in Radix bupleuri and in pharmaceuticals of the chinese multiherb remedy xiaochaihu-tang using liquid chromatographic tandem mass spectrometry.[Pubmed:15253665]

Anal Chem. 2004 Jul 15;76(14):4208-16.

Saikosaponins are bioactive oleanane saponins derived from the Chinese medicinal herb Radix bupleuri ("chaihu" in Chinese). An LC-MS/MS-based method has been developed for characterization and quantification of 15 saikosaponin derivatives (saikosaponin a, saikosaponin b(1), saikosaponin g, Saikogenin A, saikogenin H, saikosaponin c, saikosaponin h, saikosaponin i, prosaikogenin C(2), prosaikogenin B(2), saikogenin C, saikogenin B, saikosaponin d, saikosaponin b(2), and saikogenin D) in one chromatographic run. Optimization of the ionization process was performed with electrospray and atmospheric pressure chemical ionization techniques in both positive and negative ion modes. Negative ion ESI was adopted for generation of the precursor deprotonated molecules to achieve the best ionization sensitivity for the analytes. In addition, the most abundant fragment ion was chosen for each analyte to give the best CID sensitivity. Because some of the saponin derivatives are isomeric, complete resolution for the whole analytes was achieved both chromatographically and mass spectroscopically. Furthermore, optimal internal standard was successfully discovered for determination of the analytes by making use of a combinatorial chemistry approach. Good linearity over the range approximately 1.65 or 4.98 to 1200 ng/mL for the analytes was observed. The intraday accuracy and precision at nominal low, intermediate, and high concentration varied between 0.8 and 11.8% and between 80 and 116%, respectively, whereas those for interday assay were between 1.1 and 15.5% and between 86 and 119%, respectively. The lower limits of quantitation for the test compounds were approximately 16.5 to 49.4 pg on-column. The new method offered higher sensitivity and greater specificity than previously reported LC methods. After the validation, the applicability of the method for determination of these chemicals present in a variety of crude chaihu roots and in different brands of the Chinese multiherb remedy Xiaochaihu-tang (or Shosaiko-to) extract granules has been demonstrated. The sensitivity and specificity of the technique will be the basis of a method for the accurate quantification of the saikosaponin derivatives in biomatrixes.

Metabolism and pharmacokinetics of orally administered saikosaponin b1 in conventional, germ-free and Eubacterium sp. A-44-infected gnotobiote rats.[Pubmed:9657043]

Biol Pharm Bull. 1998 Jun;21(6):588-93.

The metabolic fate of saikosaponin b1 (1) was investigated using conventional, germ-free and Eubacterium sp. A-44-infected gnotobiote rats. After the oral administration of 1 to germ-free rats at a dose of 50 mg/kg, no metabolite was detected in the plasma, the cecal contents or the cumulative feces through the experiment. On the other hand, when 1 was orally given to the Eubacterium sp. A-44-infected gnotobiote rats, considerable amounts of its metabolites, proSaikogenin A (2) and Saikogenin A (3), were detected in the rat plasma with the respective AUC0-10 h values of 17,424 and 22,260 pmol.min/ml, similar to the case of its oral administration to conventional rats (AUC0-10 h values of 9,936 and 12,414 pmol.min/ml for 2 and 3, respectively). Furthermore, significant amounts of both metabolites were detected in the cecal contents and the cumulative feces of the gnotobiote and conventional rats, but not in those of the germ-free rats, within 10 h after the administration. Fecal and cecal activities of hydrolyzing 1 and 2 were found in the gnotobiote and conventional rats, though there were no detectable activities in the germ-free rats. Accordingly, both hydrolyzing activities in the intestinal bacteria, such as Eubacterium sp. A-44, are essential for the appearance of 2 and 3 in the rat plasma and cumulative feces, since orally administered 1 was poorly absorbed from the gastrointestinal tract.

Effects of saikosaponin metabolites on the hemolysis of red blood cells and their adsorbability on the cell membrane.[Pubmed:2632076]

Chem Pharm Bull (Tokyo). 1989 Dec;37(12):3306-10.

The hemolytic properties and the adsorbability on red blood cells of saikosaponin a, saikosaponin d and 13 metabolites formed in the alimentary tract were investigated. Among these compounds, saikosaponin d and its intestinal product, prosaikogenin G, which possess an alpha-hydroxyl function at C16, showed the strongest hemolytic activity at the dose range of 1.0 to 5.0 micrograms/ml. Saikosaponin a and its intestinal product, prosaikogenin F, which possess a beta-hydroxyl function at C16, showed activity above 10 micrograms/ml. In this case, the monoglycoside, prosaikogenin F, showed the stronger activity than the diglycoside, saikosaponin a. Among the gastric products whose ether ring was cleaved to produce a carbinol, the monoglycosides, proSaikogenin A and prosaikogenin H, showed a slight activity above 25 micrograms/ml, and the saikogenins except Saikogenin A were inactive. Saikogenin A, however, had hemolytic activity at a dose of 15 micrograms/ml. The adsorbabilities of these compounds on red blood cell membranes closely paralleled their degrees of hemolytic activity.

Corticosterone secretion-inducing activity of saikosaponin metabolites formed in the alimentary tract.[Pubmed:2611932]

Chem Pharm Bull (Tokyo). 1989 Oct;37(10):2736-40.

The corticosterone secretion-inducing activities of saikosaponin a, saikosaponin c and saikosaponin d, isolated from the root of Bupleurum falcatum L., and 27 metabolites formed in the murine alimentary tract were studied in mice. Serum corticosterone was determined by high-performance liquid chromatography (HPLC). Intraperitoneal administration of saikosaponin a and its intestinal metabolite, prosaikogenin F, showed corticosterone secretion-inducing activity at a dose of 0.1 mmol/kg, and maximally increased it at a dose of 0.4 mmol/kg. On the other hand, the genuine sapogenin, saikogenin F, was inactive. Saikosaponin b1 and saikosaponin g, gastric metabolites of saikosaponin a, and their intestinal metabolites, proSaikogenin A, prosaikogenin H, Saikogenin A and saikogenin H, were also inactive. Serum corticosterone was increased by the administration of saikosaponin d and its intestinal metabolite, prosaikogenin G, at a dose of 0.04 mmol/kg, and it reached the maximal level at the dose of 0.1 mmol/kg. Saikogenin G also showed a slight activity. A gastric metabolite of saikosaponin d, saikosaponin b2, and its intestinal metabolites, prosaikogenin D and saikogenin D, were inactive. In the experiments on saikosaponin c and its metabolites, saikosaponin c was inactive but its intestinal metabolites, especially prosaikogenin E-2, showed activity almost equal to that of saikosaponin a. Saikosaponin h and saikosaponin i, gastric metabolites of saikosaponin c, were also inactive, but their prosaikogenins showed slight activities. When these compounds were orally administered, their corticosterone secretion-inducing activities were similar to those obtained in the intraperitoneal experiment. These results suggest that a proper polar balance between the sugar moiety and the aglycone is important for the corticosterone secretion-inducing activity of saikosaponins and their metabolites.

Anti-inflammatory effect of saikogenin A.[Pubmed:2427082]

Biochem Pharmacol. 1986 Aug 1;35(15):2483-7.

Saikogenin A, an anti-inflammatory drug, is present in the crude extract of a Chinese herbal plant called Tsai-Fu. Saikogenin A was less effective in adrenalectomized rats than in normal rats in reducing the carrageenin-induced edema. Serum corticosterone and ACTH were increased in the Saikogenin A-treated rats, supporting the view that stimulation of hypothalamopituitary-adrenal system is responsible for the anti-inflammatory effect of Saikogenin A. This is further supported by the findings that Saikogenin A did not affect the spontaneous release of corticosterone but it facilitated the ACTH-induced release. In addition, cyclic AMP in isolated pituitary and adrenal glands was increased by Saikogenin A. A role for cyclic AMP as the second messenger is thus considered. Otherwise, the direct action of Saikogenin A on the process of inflammation cannot be ruled out because Saikogenin A also functioned in the adrenalectomized rats and it inhibited the release of histamine induced by compound 48/80. Reduction of the vascular permeability was also observed in the Saikogenin A-treated rats. These results suggest that the anti-inflammatory action of Saikogenin A are due to an increase in corticosterone caused by the release of ACTH and a direct effect on the process of inflammation.

Structural transformation of saikosaponins by gastric juice and intestinal flora.[Pubmed:4087133]

J Pharmacobiodyn. 1985 Sep;8(9):718-25.

Structural transformation of saikosaponin a and saikosaponin d, main components of Bupleuri Radix, were investigated using rat gastric juice (pH 1.5) and mouse intestinal flora in vitro and the excretion of saikosaponin derivatives in rat feces was also studied. Quantitative analysis of saikosaponins and their derivatives was carried out by high performance liquid chromatography. By the incubation of saikosaponins in rat gastric juice, saikosaponin a decreased with time dependently. After 3 h, saikosaponin a disappeared completely and saikosaponin b1 which possessed heteroannular diene moiety at C-11,13(18) and saikosaponin g which possessed homoannular diene moiety at C-9(11),12 were detected with the ratio of 3:1. On the other hand, saikosaponin d rapidly changed to only saikosaponin b2 (heteroannular diene structure) completely 30 min after the incubation. Next, by the anaerobic incubation of saikosaponin a with mouse intestinal flora, the formation of saikogenin F, a genuine aglycone of saikosaponin a, reached to the maximum 1 h after the incubation and its yield was 80%. A minor peak of prosaikogenen F, a monofucoside of saikogenin F, was also detected at 15 min. By the same procedures, saikosaponin b1, g, d and b2 also changed to the corresponded proSaikogenin A, H, G and D and Saikogenin A, H, G and D with the almost similar pattern to that of saikosaponin a. Finally, the contents of nine excreted metabolites from saikosaponin a, 5 and 20 mg/kg, in feces after its oral administration was investigated using fasted or non-fasted rats.(ABSTRACT TRUNCATED AT 250 WORDS)