| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 100mg |
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| 250mg | |||
| Other Sizes |
| Targets |
AKT/mTOR signaling pathway (HepG2 cell IC₅₀ = 25.3 μM; SMMC-7721 cell IC₅₀ = 21.7 μM) [1]
MAPK signaling pathway (ERK1/2, JNK, p38) [1] DNA damage response (γ-H2AX) [1] |
|---|---|
| ln Vitro |
Rotundic acid exhibits potent antiproliferative activity against human hepatocellular carcinoma (HCC) cells, with IC₅₀ values of 25.3 μM (HepG2) and 21.7 μM (SMMC-7721) after 48 hours of treatment (MTT assay). [1]
It induces significant DNA damage in HCC cells: Comet assay shows a dose-dependent increase in tail moment (1.8-fold and 2.5-fold higher than control at 20 μM and 40 μM, respectively), and Western blot confirms upregulated expression of γ-H2AX (a marker of DNA double-strand breaks). [1] Rotundic acid promotes HCC cell apoptosis in a dose-dependent manner: Annexin V-FITC/PI staining reveals that 40 μM treatment increases the apoptotic rate to 38.6% (HepG2) and 42.3% (SMMC-7721) after 48 hours; it also activates caspase-3 and caspase-9, and induces cleavage of PARP, as evidenced by Western blot. [1] It inhibits the AKT/mTOR pathway: Treatment with Rotundic acid (10-40 μM) dose-dependently reduces the phosphorylation levels of AKT (p-AKT) and mTOR (p-mTOR) without affecting total AKT/mTOR protein expression. [1] It modulates the MAPK pathway: Rotundic acid decreases phosphorylated ERK1/2 (p-ERK1/2) levels while increasing phosphorylated JNK (p-JNK) and phosphorylated p38 (p-p38) levels in a dose-dependent manner. [1] Rotundic acid suppresses the clonogenic potential of HCC cells: Colony formation assay shows that 20 μM and 40 μM treatments reduce the number of colonies by 56.2% and 78.5% (HepG2), and 61.3% and 82.1% (SMMC-7721), respectively, compared to the control group. [1] It has no significant cytotoxicity against normal human hepatocytes (LO2) at concentrations up to 40 μM (cell viability > 85%). [1] |
| ln Vivo |
In a HepG2 xenograft nude mouse model, intraperitoneal administration of Rotundic acid (20 mg/kg and 40 mg/kg, 3 times/week for 4 weeks) significantly inhibits tumor growth: Tumor volume is reduced by 45.8% (20 mg/kg) and 68.3% (40 mg/kg), and tumor weight is decreased by 42.5% (20 mg/kg) and 65.7% (40 mg/kg) compared to the vehicle control group (P < 0.05). [1]
Tumor tissue analysis confirms that Rotundic acid induces DNA damage (upregulated γ-H2AX expression) and apoptosis (increased TUNEL-positive cells) in vivo. [1] It modulates signaling pathways in tumor tissues: Western blot and immunohistochemistry show reduced p-AKT/p-mTOR levels and increased p-JNK/p-p38 levels in tumor tissues from treated mice. [1] |
| Cell Assay |
MTT cell viability assay: HepG2, SMMC-7721, and LO2 cells are seeded in 96-well plates (5×10³ cells/well) and cultured overnight. Rotundic acid is added at concentrations of 0, 5, 10, 20, 40, 80 μM, and cells are incubated for 48 hours. MTT reagent is added, and after 4 hours of incubation, the absorbance at 570 nm is measured to calculate cell viability and IC₅₀ values. [1]
Colony formation assay: HCC cells are seeded in 6-well plates (1×10³ cells/well) and cultured for 24 hours. Rotundic acid (0, 20, 40 μM) is added, and cells are cultured for 14 days. Colonies are fixed, stained, and counted to evaluate clonogenic potential. [1] Comet assay for DNA damage: HepG2 cells are treated with Rotundic acid (0, 20, 40 μM) for 24 hours. Cells are embedded in agarose gel, subjected to electrophoresis, stained with a DNA-binding dye, and observed under a fluorescence microscope. Tail moment is quantified to assess DNA damage. [1] Apoptosis detection: HCC cells are treated with Rotundic acid (0, 20, 40 μM) for 48 hours, stained with Annexin V-FITC and PI, and analyzed by flow cytometry to determine the apoptotic rate. [1] Western blot assay: HCC cells are treated with Rotundic acid (0, 10, 20, 40 μM) for 24 hours. Total proteins are extracted, separated by SDS-PAGE, transferred to membranes, and probed with antibodies against γ-H2AX, AKT, p-AKT, mTOR, p-mTOR, ERK1/2, p-ERK1/2, JNK, p-JNK, p38, p-p38, caspase-3, caspase-9, PARP, and cleaved PARP. GAPDH is used as an internal reference. [1] Immunofluorescence staining for γ-H2AX: HepG2 cells are seeded on coverslips, treated with Rotundic acid (0, 20, 40 μM) for 24 hours, fixed, permeabilized, and incubated with anti-γ-H2AX antibody and fluorescent secondary antibody. Nuclei are stained with DAPI, and γ-H2AX foci are observed under a confocal microscope. [1] |
| Animal Protocol |
HepG2 xenograft nude mouse model: Male BALB/c nude mice (6-8 weeks old) are subcutaneously injected with 5×10⁶ HepG2 cells into the right axilla. When tumors reach a volume of ~100 mm³, mice are randomly divided into 3 groups (n=6/group): vehicle control group (0.1% DMSO in normal saline), low-dose Rotundic acid group (20 mg/kg), and high-dose Rotundic acid group (40 mg/kg). The drug is administered via intraperitoneal injection 3 times a week for 4 weeks. Tumor volume (measured every 3 days) and body weight (measured weekly) are recorded. At the end of the experiment, mice are euthanized, tumors are excised, weighed, and fixed in 4% paraformaldehyde for histological and immunohistochemical analysis. [1]
Tumor tissue analysis: Paraffin-embedded tumor sections are subjected to HE staining (to observe histological changes) and TUNEL staining (to detect apoptotic cells). Immunohistochemistry is performed to detect the expression of p-AKT, p-mTOR, p-JNK, p-p38, and γ-H2AX in tumor tissues. [1] |
| Toxicity/Toxicokinetics |
In in vivo studies, mice treated with intraperitoneal injection of Rotundic acid (20-40 mg/kg) for 4 weeks showed no significant changes in body weight, food intake, or general behavior. [1] Serum biochemical analysis showed no significant abnormalities in liver function (ALT, AST) or kidney function (BUN, CREA) compared to the control group. [1] Histopathological examination of major organs (liver, kidney, heart, lung, spleen) showed no obvious toxic lesions in the Rotundic acid treatment group. [1]
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| References | |
| Additional Infomation |
Rotundic acid is a triterpenoid compound that acts as a metabolite. It has been reported to be found in Enkianthus campanulatus, Mussaenda macrophylla, and other organisms with relevant data. Rotundic acid is a naturally occurring triterpenoid compound isolated from medicinal plants such as Clematis chinensis Osbeck. [1] It exerts its anti-hepatocellular carcinoma effect through a dual mechanism: inhibiting the AKT/mTOR pathway (promoting cell survival and proliferation) and regulating the MAPK pathway (mediating stress response and apoptosis), ultimately inducing DNA damage and caspase-dependent cell death. [1] The selective cytotoxicity of Rotundic acid to hepatocellular carcinoma cells (with minimal toxicity to normal hepatocytes) and its good in vivo safety make it a potential lead compound for the development of therapeutics for hepatocellular carcinoma. [1]
|
| Molecular Formula |
C30H48O5
|
|---|---|
| Molecular Weight |
488.6991
|
| Exact Mass |
488.35
|
| CAS # |
20137-37-5
|
| PubChem CID |
12315075
|
| Appearance |
White to off-white solid powder
|
| Density |
1.2±0.1 g/cm3
|
| Boiling Point |
622.8±55.0 °C at 760 mmHg
|
| Melting Point |
272-274℃
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| Flash Point |
344.5±28.0 °C
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| Vapour Pressure |
0.0±4.1 mmHg at 25°C
|
| Index of Refraction |
1.581
|
| LogP |
5.75
|
| Hydrogen Bond Donor Count |
4
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
2
|
| Heavy Atom Count |
35
|
| Complexity |
945
|
| Defined Atom Stereocenter Count |
11
|
| SMILES |
C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H]([C@@]5(C)CO)O)C)C)[C@@H]2[C@]1(C)O)C)C(=O)O
|
| InChi Key |
YLHQFGOOMKJFLP-LTFXOGOQSA-N
|
| InChi Code |
InChI=1S/C30H48O5/c1-18-9-14-30(24(33)34)16-15-27(4)19(23(30)29(18,6)35)7-8-21-25(2)12-11-22(32)26(3,17-31)20(25)10-13-28(21,27)5/h7,18,20-23,31-32,35H,8-17H2,1-6H3,(H,33,34)/t18-,20-,21-,22+,23-,25+,26+,27-,28-,29-,30+/m1/s1
|
| Chemical Name |
(1R,2R,4aS,6aR,6aS,6bR,8aR,9R,10S,12aR,14bS)-1,10-dihydroxy-9-(hydroxymethyl)-1,2,6a,6b,9,12a-hexamethyl-2,3,4,5,6,6a,7,8,8a,10,11,12,13,14b-tetradecahydropicene-4a-carboxylic acid
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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)
|
| Solubility (In Vitro) |
DMSO : ~100 mg/mL (~204.62 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (5.12 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
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. Solubility in Formulation 2: 2.5 mg/mL (5.12 mM) 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; with ultrasonication. 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (5.12 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 | 2.0462 mL | 10.2312 mL | 20.4625 mL | |
| 5 mM | 0.4092 mL | 2.0462 mL | 4.0925 mL | |
| 10 mM | 0.2046 mL | 1.0231 mL | 2.0462 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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