Ginsenoside Rh2, a naturally occurring steroid glycoside and an aldose reductase inhibitor, is isolated from the root of ginseng and has a variety of biological effects. Caspase-8 and Caspase-9 activation can be induced by ginsenoside Rh2. Ginsenoside Rh2 induces apoptosis in cancer cells via multiple pathways. In Alzheimer's disease models, it also lowers amyloid-β levels, induces β -cell proliferation, and inhibits osteoclastogenesis by stifling RANKL-induced osteoclast differentiation.
Physicochemical Properties
| Molecular Formula | C36H62O8 |
| Molecular Weight | 622.8727 |
| Exact Mass | 622.444 |
| Elemental Analysis | C, 69.42; H, 10.03; O, 20.55 |
| CAS # | 78214-33-2 |
| Related CAS # | 78214-33-2 |
| PubChem CID | 119307 |
| Appearance | White to off-white solid powder |
| Density | 1.2±0.1 g/cm3 |
| Boiling Point | 726.4±60.0 °C at 760 mmHg |
| Flash Point | 393.1±32.9 °C |
| Vapour Pressure | 0.0±5.4 mmHg at 25°C |
| Index of Refraction | 1.572 |
| LogP | 5.62 |
| Hydrogen Bond Donor Count | 6 |
| Hydrogen Bond Acceptor Count | 8 |
| Rotatable Bond Count | 7 |
| Heavy Atom Count | 44 |
| Complexity | 1070 |
| Defined Atom Stereocenter Count | 15 |
| SMILES | O([H])C1([H])C([H])([H])C2([H])[C@@]3(C([H])([H])[H])C([H])([H])C([H])([H])C([H])(C(C([H])([H])[H])(C([H])([H])[H])C3([H])C([H])([H])C([H])([H])[C@@]2(C([H])([H])[H])[C@]2(C([H])([H])[H])C([H])([H])C([H])([H])C([H])([C@](C([H])([H])[H])(C([H])([H])C([H])([H])/C(/[H])=C(\C([H])([H])[H])/C([H])([H])[H])O[H])C21[H])OC1([H])C([H])(C([H])(C([H])(C([H])(C([H])([H])O[H])O1)O[H])O[H])O[H] |
| InChi Key | CKUVNOCSBYYHIS-IRFFNABBSA-N |
| InChi Code | InChI=1S/C36H62O8/c1-20(2)10-9-14-36(8,42)21-11-16-35(7)27(21)22(38)18-25-33(5)15-13-26(32(3,4)24(33)12-17-34(25,35)6)44-31-30(41)29(40)28(39)23(19-37)43-31/h10,21-31,37-42H,9,11-19H2,1-8H3/t21-,22+,23+,24-,25+,26-,27-,28+,29-,30+,31-,33-,34+,35+,36-/m0/s1 |
| Chemical Name | (2R,3R,4S,5S,6R)-2-[[(3S,5R,8R,9R,10R,12R,13R,14R,17S)-12-hydroxy-17-[(2S)-2-hydroxy-6-methylhept-5-en-2-yl]-4,4,8,10,14-pentamethyl-2,3,5,6,7,9,11,12,13,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl]oxy]-6-(hydroxymethyl)oxane-3,4,5-triol |
| Synonyms | 20(S)-Ginsenoside Rh2; 20(S)-Rh2; Ginsenoside-Rh2;AR-1A4936; AR1A4936; AR 1A4936 |
| HS Tariff Code | 2934.99.9001 |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets | Caspase-8; Caspase-9; Apoptosis |
| ln Vitro | Ginsenoside Rh2 induces the activation of two initiator caspases, caspase-8 and caspase-9 in human cancer cells. Ginsenoside Rh2 is a promising candidate for the development of anti-tumor medications since it induces cancer cells to die via multiple pathways. Ginsenoside Rh2 initiates p53-dependent Fas expression, which leads to the activation of caspase-8 and p53-independent caspase-9-mediated intrinsic pathway, ultimately leading to cancer cell death. The human tumor cell lines HeLa, SK-HEP-1, SW480, and PC-3 are used to determine Ginsenoside Rh2's cytotoxic activity. SK-HEP-1 and SW480 cells are less sensitive to Ginsenoside Rh2, with IC50 values of 3.15 g/mL and 4.06 μg/mL, respectively, while HeLa cells are significantly inhibited in their ability to divide, with an IC50 value of 2.52 μg/mL. With an IC50 value of 7.85 μg/mL, 3-fold higher than HeLa cells, PC-3 cells are the least susceptible to Ginsenoside Rh2[1]. |
| ln Vivo | Tumor sizes from the three groups that are tumor-bearing are measured 15 days after B16-F10 cells were injected. In comparison to the tumor group, the tumor sizes in the G-L and G-H groups (where G-L and G-H denote a low or high dose of ginsenoside Rh2 injection, respectively) are smaller (P<0.05). The Ginsenoside Rh2 treated groups outlive the tumor group that is not receiving treatment, and the effect is dose-dependent (P<0.05), according to the survival analysis [2]. |
| Enzyme Assay | Ginsenoside Rh2 (7.5 μg/mL) is applied to HeLa, SK-HEP-1, SW480, and PC-3 cells in serum-free media for the indicated times before the cells are harvested. 50 micrograms of cell lysates are incubated with 200 nM Ac-DEVD-AFC, Ac-IETD-AFC, and Ac-LEHD-AFC for 1 hour at 37°C in a reaction buffer containing 20 mM HEPES, pH 7.4, 100 mM NaCl, 10 mM DTT, 0.1% CHAPS, and 10% sucrose. Fluorescence emission at 535 nm and excitation at 405 nm are used to monitor the reaction[1]. |
| Cell Assay | Determination of cell viability is performed by using MTT assay, which is used to calculate the growth inhibition induced by increasing concentrations of drug. In a 96-well plate, 1104 HeLa, SK-HEP-1, SW480, and PC-3 cells that are exponentially growing are seeded in triplicate. Cells are treated with increasing concentrations of ginsenoside Rh2 (1, 2.5, 5, 7.5, and 10 μg/mL) in serum-free media for 48 hours after 24 hours of incubation. Each well receives 20 μL of MTT (5 mg/mL) at the conclusion of the treatment, which is then incubated for an additional 4 hours. The DMSO is used to solubilize the formazan grains produced by viable cells, and an ELISA reader is used to measure the color intensity at 550 nm[1]. |
| Animal Protocol | Mice: In 4 groups of 80 mice each, the Tumor group, G-L group, G-H group, and Control group of male C57BL6 mice (3–4 weeks old) are randomly assigned. A low or high dose of ginsenoside Rh2 is referred to as G-L or G-H. The B16-F10 cell line is administered to the mice in the tumor group, G-L group, and G-H group. These 3 groups develop into cancer-bearing groups. The identical volume of PBS is injected in the control group in place of the drug. In the G-L and G-H groups of mice, ginsenoside Rh2 is injected into the left back. After day 5, the dose for the G-H group is 0.5 mg/kg or 0.2 mg/kg for the G-L group. PBS is injected into the tumor and control groups at the same time points. |
| References |
[1]. p53-dependent Fas expression is critical for Ginsenoside Rh2 triggered caspase-8 activation in HeLa cells. Protein Cell. 2014 Mar;5(3):224-34. [2]. Ginsenoside Rh2 enhances the antitumor immunological response of a melanoma mice model. Oncol Lett. 2017 Feb;13(2):681-685. |
| Additional Infomation |
(20S)-ginsenoside Rh2 is a ginsenoside found in Panax species that is dammarane which is substituted by hydroxy groups at the 3beta, 12beta and 20 pro-S positions, in which the hydroxy group at position 3 has been converted to the corresponding beta-D-glucopyranoside, and in which a double bond has been introduced at the 24-25 position. It has a role as a plant metabolite, an antineoplastic agent, an apoptosis inducer, a cardioprotective agent, a bone density conservation agent and a hepatoprotective agent. It is a beta-D-glucoside, a 12beta-hydroxy steroid, a ginsenoside, a tetracyclic triterpenoid and a 20-hydroxy steroid. It derives from a hydride of a dammarane. Ginsenoside Rh2 has been reported in Panax notoginseng and Panax ginseng with data available. |
Solubility Data
| Solubility (In Vitro) |
DMSO: 50~100 mg/mL (80.3~160.6 mM) Ethanol: ~100 mg/mL (~160.6 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.25 mg/mL (2.01 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 12.5 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: ≥ 1.25 mg/mL (2.01 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), suspension solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.6055 mL | 8.0274 mL | 16.0547 mL | |
| 5 mM | 0.3211 mL | 1.6055 mL | 3.2109 mL | |
| 10 mM | 0.1605 mL | 0.8027 mL | 1.6055 mL |