- Hederasaponin B exhibited antiviral activity against EV71 C3 with an EC₅₀ of 24.77 ± 12.56 μg/ml and against EV71 C4a with an EC₅₀ of 41.77 ± 0.76 μg/ml, as determined by the sulforhodamine B (SRB) cytopathic effect (CPE) reduction method. The therapeutic index (TI = CC₅₀/EC₅₀) was 2.02 for EV71 C3 and 1.18 for EV71 C4a. [1]

- In contrast, ribavirin (used as a positive control) showed no significant antiviral activity against either EV71 subgenotype, as it did not reduce CPE or viral protein expression. [1]

- Western blot analysis revealed that treatment with hederasaponin B at 50 μg/ml dramatically decreased the expression of viral capsid protein VP2 (34 kDa) in vero cells infected with EV71 C3 or EV71 C4a at 48 h post‑infection, indicating inhibition of viral capsid protein synthesis. [1]

- Microscopic observation showed that hederasaponin B (50 μg/ml) prevented the formation of visible CPE induced by EV71 C3 or EV71 C4a, whereas ribavirin did not protect cells from virus‑induced CPE. [1]

- Antiviral activity assay (CPE reduction by SRB method): Vero cells were seeded in 96‑well plates at 2×10⁴ cells/well one day before infection. The next day, the medium was removed, cells were washed with PBS, and 0.09 ml of diluted virus suspension (containing 50% cell culture infective dose, CCD₅₀) was added to each well to induce CPE within 48 h. Then 0.01 ml of medium supplemented with FBS containing serial 5‑fold dilutions of hederasaponin B (0.4 to 50 μg/ml) was added. Virus‑infected non‑treated cells and non‑infected non‑treated cells served as controls. After 2 days of incubation at 37 °C in 5% CO₂, cells were fixed with cold 70% acetone (−20 °C) for 30 min, dried, stained with 0.4% SRB in 1% acetic acid for 30 min at room temperature, washed 5 times with 1% acetic acid, and dried. Bound SRB was solubilized with 10 mM Tris‑base solution (100 μl), and absorbance was read at 540 nm (reference 620 nm). Percent protection was calculated as: (OD_EV71 treated − OD_EV71 control) / (OD_mock − OD_EV71 control) × 100. [1]

- Cytotoxicity assay: Vero cells were seeded in 96‑well plates at 2×10⁴ cells/well. After 24 h, cells were treated with various concentrations of hederasaponin B in maintenance medium for 48 h at 37 °C in parallel with the virus‑infected cultures. Cytotoxicity was evaluated by the same SRB assay described above, and the CC₅₀ (concentration reducing cell growth by 50%) was determined. For hederasaponin B, the CC₅₀ was >50 μg/ml against both EV71 C3 and C4a, indicating no significant cytotoxicity at the tested concentrations. [1]

- Western blot analysis for VP2 protein: Vero cells were plated in 6‑well plates at 5×10⁶ cells/well 24 h before infection with EV71 C3 or EV71 C4a. Infected cells were treated with hederasaponin B or ribavirin at 50 μg/ml for 48 h. Mock‑infected cells (0.1% DMSO) and virus‑infected cells (0.1% DMSO) were used as controls. Cells were lysed in ice‑cold RIPA buffer (containing Tris‑HCl pH 7.4, NaCl, NP‑40, sodium deoxycholate, SDS, EDTA, aprotinin, leupeptin, PMSF, sodium fluoride, and sodium orthovanadate). Protein samples (30 μg) were boiled for 10 min at 100 °C, separated on 12% acrylamide gels at 100 V for 1 h, and transferred to nitrocellulose membranes using a transfer device at 20 V for 7 min. Membranes were blocked with 5% skim milk in PBST overnight at 4 °C, then incubated with primary mouse anti‑enterovirus 71 monoclonal antibody (1:1,000 dilution) for VP2 detection, or with primary α‑tubulin mouse monoclonal IgG1 (1:1,000) as a loading control. After washing with PBST, membranes were incubated with HRP‑conjugated goat anti‑mouse secondary antibody (1:5,000) for 1 h at room temperature. Protein bands were visualized by enhanced chemiluminescence (ECL). [1]

- The CC₅₀ of hederasaponin B in vero cells was >50 μg/ml, indicating that at concentrations up to 50 μg/ml, the compound did not reduce cell viability and showed no signs of cytotoxicity as assessed by SRB assay and microscopic observation of cell morphology. [1]

[1]. Antiviral Activity of Hederasaponin B from Hedera helix against Enterovirus 71 Subgenotypes C3 and C4a. Biomol Ther (Seoul). 2014 Jan;22(1):41-6.

According to reports, ivy saponin B is present in anemones, purple leaf algae, and other organisms for which data are available.

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- Hederasaponin B is a triterpenoid saponin. Previous studies have suggested that triterpenoid saponins may inhibit viral nucleotide synthesis (e.g., against herpes simplex virus type‑1), and the authors hypothesize that hederasaponin B may exert its anti‑EV71 effect through a similar mechanism, suppressing enterovirus replication. [1]

- The 30% ethanol extract of Hedera helix containing hederasaponin B is already clinically used as a safe medicine for the treatment of bronchitis in children. The extract also showed significant antiviral activity against EV71 C3 (EC₅₀ = 6.58 μg/ml, TI = 7.59) and EV71 C4a (EC₅₀ = 22.00 μg/ml, TI = 4.55) in vitro, suggesting that the extract itself could be developed as an anti‑EV71 therapeutic. [1]

- No approved antiviral drug currently exists for enterovirus 71 infections. Hederasaponin B represents a potential broad‑spectrum antiviral candidate effective against multiple EV71 subgenotypes (C3 and C4a). [1]

C59H96O25

1205.3786

1204.624

C, 58.79; H, 8.03; O, 33.18

36284-77-2

21626480

White to off-white solid powder

1.5±0.1 g/cm3

223-226 °C

1.630

6.11

14

25

13

84

2350

32

O(C1([H])C([H])(C([H])(C([H])(C([H])([H])O1)O[H])O[H])OC1([H])C([H])(C([H])(C([H])(C([H])(C([H])([H])[H])O1)O[H])O[H])O[H])C1([H])C([H])([H])C([H])([H])C2(C([H])([H])[H])C([H])(C([H])([H])C([H])([H])C3(C([H])([H])[H])C4(C([H])([H])[H])C([H])([H])C([H])([H])C5(C(=O)OC6([H])C([H])(C([H])(C([H])(C([H])(C([H])([H])OC7([H])C([H])(C([H])(C([H])(C([H])(C([H])([H])O[H])O7)OC7([H])C([H])(C([H])(C([H])(C([H])(C([H])([H])[H])O7)O[H])O[H])O[H])O[H])O[H])O6)O[H])O[H])O[H])C([H])([H])C([H])([H])C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])C5([H])C4=C([H])C([H])([H])C32[H])C1(C([H])([H])[H])C([H])([H])[H]

NVSLBOBPSCMMSO-BVLVEXITSA-N

InChI=1S/C59H96O25/c1-24-34(62)38(66)42(70)49(77-24)82-46-29(21-60)79-48(45(73)41(46)69)76-23-30-37(65)40(68)44(72)51(80-30)84-53(74)59-18-16-54(3,4)20-27(59)26-10-11-32-56(7)14-13-33(55(5,6)31(56)12-15-58(32,9)57(26,8)17-19-59)81-52-47(36(64)28(61)22-75-52)83-50-43(71)39(67)35(63)25(2)78-50/h10,24-25,27-52,60-73H,11-23H2,1-9H3/t24-,25-,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-,56-,57+,58+,59-/m0/s1

[(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] (4aS,6aR,6aS,6bR,8aR,10S,12aR,14bS)-10-[(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-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,7,8,8a,10,11,12,13,14b-tetradecahydropicene-4a-carboxylate

Hederagenin B; Hederacoside B

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~100 mg/mL (~82.96 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (2.07 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 25.0 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.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (2.07 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 25.0 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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (2.07 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)