| Size | Price | Stock | Qty |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg | |||
| 500mg | |||
| Other Sizes |
| ln Vitro |
Pulchinenoside C (Anemoside B4) can suppress the release of IL-10; SSA, SSD and PNS upregulate IL-2 expression [1]. Pulchinenoside C (Anemoside B4), IC50 value is larger than 390 ug/mL. Anemomoside B4 suppresses cell proliferation [2]. Pulchinenoside C (Anemoside B4) can greatly reduce the release of the inflammatory factor E-selectin by endothelial cells [3].
Pulchinenoside A (PA) exhibited cytotoxic activity against human chronic myelogenous leukemia K562 cells with an IC₅₀ of 103.7 ± 9.8 μg/mL (138.2 ± 13 μM). It also showed activity against mouse melanoma B16 cells (IC₅₀ provided in the table). 23-Hydroxybetulinic acid (HBA) was the most cytotoxic compound against K562 cells (IC₅₀ = 18.7 ± 1.3 μg/mL or 39.9 ± 2.7 μM) and other cell lines (B16, HeLa, HUVEC). Anemoside B4 (AB4) showed very low cytotoxicity (IC₅₀ > 390 μg/mL or >320 μM against K562). HBA induced S-phase cell cycle arrest and apoptosis in K562 cells in a concentration-dependent manner. HBA treatment (20, 40, 80 μM for 24h) significantly disrupted mitochondrial membrane potential (MMP), increased the pro-apoptotic protein Bax, decreased the anti-apoptotic proteins Bcl-2 and survivin, promoted cytochrome C release from mitochondria, and activated cleaved caspase-9 and caspase-3, indicating induction of the intrinsic apoptosis pathway.[2] |
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| Cell Assay |
Cell Viability (MTT Assay): Cells (K562, B16, HeLa, HUVEC) were seeded in 96-well plates (4000 cells/well), cultured for 24h, then treated with various concentrations of total saponins (TPS, 0-200 μg/mL) or the pure compounds (HBA, PA, AB4, 0-300 μM) for 48h. MTT solution (0.5 mg/mL) was added and incubated for 4h. The formed formazan crystals were dissolved in DMSO, and absorbance was measured at 490 nm. IC₅₀ values were determined from concentration-response curves.
Cell Cycle Analysis: K562 cells were treated with HBA (0-80 μM) for 24h, fixed in ice-cold 70% ethanol, treated with RNase, stained with propidium iodide (PI), and analyzed by flow cytometry to determine DNA content and cell cycle distribution. Apoptosis Detection (Hoechst33342/PI Staining): K562 cells treated with HBA (0-80 μM) for 24h were stained with Hoechst33342 and PI, then observed under a fluorescence microscope to assess nuclear condensation (bright blue) and necrosis/late apoptosis (red). Apoptosis Detection (Annexin V-FITC/PI by Flow Cytometry): K562 cells treated with HBA (0-80 μM) for 24h were stained with Annexin V-FITC and PI and immediately analyzed by flow cytometry to quantify early and late apoptotic cells. Mitochondrial Membrane Potential (MMP) Assessment: K562 cells treated with HBA (0-80 μM) for 24h were stained with JC-1 dye for 20 min at 37°C. MMP changes were analyzed by fluorescence microscope and spectrophotometer. The ratio of red (JC-1 aggregates, high MMP) to green (JC-1 monomers, low MMP) fluorescence intensity was calculated. Western Blotting: K562 cells treated with HBA (0-80 μM) for 24h were lysed. Proteins were separated by SDS-PAGE, transferred to membranes, and probed with antibodies against Bax, Bcl-2, cytochrome C, survivin, cleaved caspase-9, cleaved caspase-3, and β-actin (loading control). Proteins were detected using enhanced chemiluminescence.[2] |
| Toxicity/Toxicokinetics |
One reference reported that total saponins (TPS) extracted from Imperata cylindrica root showed no toxicity in nude mice after being administered doses of 100 and 200 mg/kg for 19 consecutive days. However, no specific toxicity data (e.g., LD₅₀, organ toxicity) were provided for the isolated compounds HBA, PA, or AB4. [2]
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| References |
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| Additional Infomation |
Anemoside B4 is a triterpenoid saponin.
This study identified Pulchinenoside A (PA) as a cytotoxic component of Pulchinenoside. Structure-activity relationship analysis showed that glycosylation of the aglycone HBA at the C-3 position (forming PA) significantly reduced cytotoxicity compared to HBA. Further glycosylation at the C-28 position (as in AB4) almost completely eliminated the activity. This suggests that the free hydroxyl groups at C-3 and C-28 are crucial for the cytotoxic activity of these triterpenoid saponins. The main mechanism of action of the most active compound, HBA, involves inducing S-phase cell cycle arrest in K562 leukemia cells and activating the mitochondrial (endogenous) apoptosis pathway. [2] |
| Molecular Formula |
C₅₉H₉₆O₂₆
|
|---|---|
| Molecular Weight |
1221.38
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| Exact Mass |
1220.619
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| CAS # |
129741-57-7
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| Related CAS # |
Pulchinenoside A;129724-84-1
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| PubChem CID |
71307558
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| Appearance |
White to light yellow solid powder
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| Density |
1.5±0.1 g/cm3
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| Melting Point |
208-215ºC
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| Index of Refraction |
1.634
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| LogP |
4.4
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| Hydrogen Bond Donor Count |
15
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| Hydrogen Bond Acceptor Count |
26
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| Rotatable Bond Count |
15
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| Heavy Atom Count |
85
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| Complexity |
2340
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| Defined Atom Stereocenter Count |
35
|
| SMILES |
C[C@H]1[C@@H]([C@H]([C@H]([C@@H](O1)O[C@@H]2[C@H](O[C@H]([C@@H]([C@H]2O)O)OC[C@@H]3[C@H]([C@@H]([C@H]([C@@H](O3)OC(=O)[C@]45CC[C@H]([C@H]4[C@H]6CC[C@@H]7[C@]8(CC[C@@H]([C@@]([C@@H]8CC[C@]7([C@@]6(CC5)C)C)(C)CO)O[C@H]9[C@@H]([C@H]([C@H](CO9)O)O)O[C@H]1[C@@H]([C@@H]([C@H]([C@@H](O1)C)O)O)O)C)C(=C)C)O)O)O)CO)O)O)O
|
| InChi Key |
OUHBKBTZUPLIIA-QEEGUAPESA-N
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| InChi Code |
InChI=1S/C59H96O26/c1-23(2)26-11-16-59(54(75)85-52-45(73)41(69)38(66)30(81-52)21-77-49-46(74)42(70)47(29(19-60)80-49)83-50-43(71)39(67)35(63)24(3)78-50)18-17-57(7)27(34(26)59)9-10-32-55(5)14-13-33(56(6,22-61)31(55)12-15-58(32,57)8)82-53-48(37(65)28(62)20-76-53)84-51-44(72)40(68)36(64)25(4)79-51/h24-53,60-74H,1,9-22H2,2-8H3/t24-,25-,26-,27+,28-,29+,30+,31+,32+,33-,34-,35-,36-,37-,38+,39+,40+,41-,42+,43+,44+,45+,46+,47+,48+,49+,50-,51-,52-,53-,55-,56-,57+,58+,59-/m0/s1
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| Chemical Name |
[(2S,3R,4S,5S,6R)-6-[[(2R,3R,4R,5S,6R)-3,4-dihydroxy-6-(hydroxymethyl)-5-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxyoxan-2-yl]oxymethyl]-3,4,5-trihydroxyoxan-2-yl] (1R,3aS,5aR,5bR,7aR,8R,9S,11aR,11bR,13aR,13bS)-9-[(2S,3R,4S,5S)-4,5-dihydroxy-3-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxyoxan-2-yl]oxy-8-(hydroxymethyl)-5a,5b,8,11a-tetramethyl-1-prop-1-en-2-yl-1,2,3,4,5,6,7,7a,9,10,11,11b,12,13,13a,13b-hexadecahydrocyclopenta[a]chrysene-3a-carboxylate
|
| Synonyms |
Pulchinenoside C Anemoside B4
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~125 mg/mL (~102.34 mM)
H2O : ~50 mg/mL (~40.94 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (1.70 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (1.70 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (1.70 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.8187 mL | 4.0937 mL | 8.1875 mL | |
| 5 mM | 0.1637 mL | 0.8187 mL | 1.6375 mL | |
| 10 mM | 0.0819 mL | 0.4094 mL | 0.8187 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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