Raddeanin A targets the ROS/JNK and NF-κB signaling pathways in osteosarcoma cells. [1]
Raddeanin A down-regulates the full-length androgen receptor (AR-FL) and its splice variants (AR-Vs) in prostate cancer cells. [3]
Raddeanin A targets the PI3K/AKT signaling pathway in colorectal cancer cells. [4]

In human osteosarcoma cell lines (MG-63, HOS, U-2 OS, SAOS-2, 143B), Raddeanin A dose- and time-dependently decreased cell viability. The IC50 values for MG-63 cells were 3.31 μM (24h) and 1.60 μM (48h); for HOS cells were 4.86 μM (24h) and 2.57 μM (48h); for U-2 OS cells were 10.05 μM (24h) and 3.91 μM (48h); for SAOS-2 cells were 5.82 μM (24h) and 3.29 μM (48h); and for 143B cells were 5.48 μM (24h) and 2.97 μM (48h). The inhibitory effect on normal bone cells (hFOB1.19) was less than on osteosarcoma cells. Raddeanin A induced mitochondria-dependent apoptosis, evidenced by increased nuclear fragmentation, apoptotic body formation, a dose-dependent increase in early and late apoptotic cells, and depolarization of mitochondrial membrane potential. It down-regulated the Bcl-2/Bax ratio and increased expression of cleaved caspase-3 and PARP. At non-toxic concentrations (0.25, 0.5, and 1 μM), Raddeanin A inhibited migration and invasion of OS cells by suppressing MMP-2/9 expression associated with NF-κB-dependent transcription. Raddeanin A significantly increased intracellular ROS levels and stimulated phosphorylation of JNK, while decreasing p-IκBα in the cytosol and p65 in the nucleus, inhibiting NF-κB transcriptional activity. [1]
In cholangiocarcinoma cell lines (RBE, LIPF155C, LIPF178C, LICCF), Raddeanin A reduced cell viability in a dose-dependent pattern. The EC50 ranges were 50.95-64.76 μg/mL and LC50 ranges were 34.65-49.47 μg/mL. At 13 μg/mL, it impaired migration and colony formation abilities in RBE and LIPF155C cells. Raddeanin A sensitized cell lines to 5-fluorouracil (5-Fu), reducing EC50 (about 2-fold) and LC50 (more than 3-fold). In RBE/5-Fu resistant cells, it promoted cell death and enhanced the effect of 5-Fu. Raddeanin A alone decreased Wee1 protein expression, while the combination with 5-Fu decreased COX-2, Bcl-2, and Wee1 but increased Bax, cyclin D1, and cyclin E. [2]
In prostate cancer cells (22Rv1, C4-2, C4-2B, LNCaP95), Raddeanin A (0-6 μmol/L) inhibited growth in a dose- and time-dependent manner, independent of androgen, but no inhibition was observed in AR-null cells (PC-3, DU145). It suppressed transcriptional activities of both AR-FL and AR-Vs, down-regulated PSA and UBE2C mRNA levels, and decreased AR-FL and AR-V protein levels. Raddeanin A induced proteasome-mediated degradation of AR-FL and AR-V proteins and suppressed transcription of the AR gene. It enhanced the growth inhibitory efficacy of docetaxel, with combination index values <1 indicating synergy. [3]
In HCT116 colorectal cancer cells, Raddeanin A (1-16 μM for 12h) inhibited proliferation with an IC50 of 2.61 μM. It induced apoptosis (14.0±0.8% at 2 μM and 26.2±0.3% at 4 μM vs. 2.2±1.1% control) and caused G0/G1 phase arrest (47.5±1.3% at 2 μM and 52.1±2.1% at 4 μM vs. 26.5±1.2% control). It increased cleaved caspase-3, cleaved PARP, and BAX expression, and decreased caspase-3, PARP, Bcl-2, cyclinD1, cyclinE, CDK4, and CDK2. Raddeanin A down-regulated p-PI3K and p-AKT proteins, and this effect was confirmed using the PI3K inhibitor LY294002. [4]

In a HOS subcutaneous xenograft model in nude mice, Raddeanin A (administered at doses of 1.25, 2.5, and 5 mg/kg) significantly decelerated tumor growth in a dose-dependent manner. Tumor weights and volumes were significantly reduced. Raddeanin A at 5 mg/kg exhibited better anti-tumor effects than doxorubicin at 5 mg/kg. TUNEL staining indicated significant apoptosis induction in Raddeanin A-treated tumors. Immunohistochemistry analysis confirmed that Raddeanin A treatment significantly enhanced p-JNK protein expression and suppressed p65 protein expression in tumor tissues. [1]
In an HCT116 xenograft model in BALB/c nude mice, Raddeanin A (4 mg/kg, injected intraperitoneally) significantly inhibited tumor growth. The average tumor volume in the control group was 2.01±0.86 cm³ and weight was 2.31±1.36 g, while in the Raddeanin A-treated group, volume was 1.01±1.36 cm³ and weight was 1.26±1.05 g. HE staining of tumor tissue showed a larger necrotic area in the treatment group (almost half) compared to control (less than one-quarter). TUNEL staining showed an apoptosis rate of 43.6±1.26% in the RA group vs. few apoptotic cells in control. Western blot and immunohistochemistry confirmed increased cleaved caspase-3, cleaved PARP, and BAX, and decreased caspase-3, PARP, Bcl-2, cyclinD1, cyclinE, CDK4, CDK2, p-PI3K, and p-AKT. No structural differences in liver tissue were observed between control and treatment groups. [4]

Cell viability was determined using an MTT assay. Human osteosarcoma cells were plated in 96-well plates and treated with various concentrations of Raddeanin A (0.2, 0.5, 1, 2, 5, 10, 20, and 50 μM) for 24 and 48 hours. MTT solution was added for 4 hours, then DMSO was added to dissolve formazan crystals, and absorbance was measured at 490 nm. [1]
Apoptosis was evaluated using an Annexin V-FITC/PI apoptosis detection kit. Cells were treated with Raddeanin A (1, 2, and 4 μM) for 24 hours, then stained, and fluorescence was detected by flow cytometry. Hoechst 33258 staining was also performed: cells treated with Raddeanin A for 12 hours were fixed, stained with Hoechst 33258 dye, and observed under a fluorescence microscope. Mitochondrial membrane potential was analyzed using JC-1 staining solution, followed by flow cytometry. [1]
Wound healing assay: cells were grown to 75% confluence, wounded with a sterile pipette tip, and treated with Raddeanin A (0.25, 0.5, and 1 μM) for 24 hours. Migration and invasion were evaluated using Transwell chambers with 8.0 μm pore size polycarbonate membrane. For invasion, Matrigel was added to the membrane. Cells were plated in the top chamber with Raddeanin A and without FBS; medium with 10% FBS was in the bottom chamber. After 24 hours, migrated/invaded cells were fixed, stained with Giemsa, and counted. [1]
For cholangiocarcinoma cells, viability was evaluated using the ATPlite assay. Cells were seeded in 96-well plates and treated with Raddeanin A (0-160 μg/mL) for 24 hours. Wound-healing migration and Transwell invasion assays were performed. Clonogenic assay: cells were treated, detached, seeded in 6-well plates (600 cells/well) in drug-free media, and colonies were stained with cristal violet and counted after 10 days. Hoechst staining was used to test apoptosis. [2]
For prostate cancer cells, growth was assessed by sulforhodamine B (SRB) assay. Cells were treated with Raddeanin A (0-6 μmol/L) in the presence or absence of 1 nmol/L R1881. Western blot analysis was performed with standard protocol using anti-AR and anti-GAPDH antibodies. DNA transfection and reporter gene assays used ARR3-luc, UBE2C-luc, and pGL4-ARpro1.7 plasmids. qRT-PCR was performed for AR-FL, AR-V7, PSA, UBE2C, and 36B4. [3]
For colorectal cancer cells, MTT assay was used. Flow cytometry for apoptosis: cells treated with Raddeanin A for 12 hours were stained with Annexin-V-FITC and PI. Cell cycle: cells were fixed with 70% ethanol, stained with PI/RNase A, and analyzed by flow cytometry. RT-PCR and western blotting were performed for various apoptosis and cell cycle-related proteins. [4]

For the osteosarcoma in vivo model, 5-week-old male BALB/c nude mice were subcutaneously injected with HOS cells. When tumors reached a certain size, mice were randomized into groups (n=6) and treated intraperitoneally with Raddeanin A (1.25, 2.5, 5 mg/kg), doxorubicin (5 mg/kg, positive drug), or vehicle (saline with 0.1% DMSO) once a day for 20 consecutive days. Tumor length and width were measured every 3 days. Tumor volume was calculated as 0.5 × length × width × thickness. At the end of treatment, mice were sacrificed, and tumor tissues were removed for histopathological, immunohistochemistry, and TUNEL assays. Normal tissues (heart, liver, spleen, lung, kidney) were also examined for toxicity. [1]
For the colorectal cancer xenograft model, HCT116 cells (1×10⁶ cells/mouse) were injected into 5-week-old BALB/c nude mice. When tumors reached approximately 150 mm³, mice were divided into two groups (n=3 per group): control (PBS 100 μL) and Raddeanin A (4 mg/kg). The route of administration was not explicitly stated as intraperitoneal or other, but the compound was injected. Tumor volume and body weight were recorded every other day. After 2 weeks, tumor tissues were removed. HE staining, TUNEL assay, immunohistochemistry, and western blot were performed on tumor and liver tissues. [4]

In the osteosarcoma xenograft study, Raddeanin A treatments were well-tolerated with no obvious systemic toxicity. No weight loss was observed during the entire treatment period. Histopathological data showed no histological alterations in low- and middle-dose groups compared to control. Slight changes in histological pattern for spleen, lungs, and kidneys were observed at the highest dose (5 mg/kg), similar to doxorubicin. No obvious pathological changes were noted in heart, liver, spleen, lung, and kidney tissues at the submicroscopic level in Raddeanin A-treated groups. [1]
In the colorectal cancer xenograft study, Raddeanin A (4 mg/kg) showed no structural differences in liver tissue between control and treatment groups by HE staining, indicating no significant hepatotoxicity. [4]
In normal bone cells (hFOB1.19), the inhibitory effect of Raddeanin A was less than that on human osteosarcoma cells at the same concentration. [1]
In cholangiocarcinoma studies, the normal intrahepatic biliary epithelial cell line HIBEpIC showed higher EC50 (79.52 μg/mL) and LC50 (63.17 μg/mL) values compared to tumor cell lines, indicating that the dose of Raddeanin A that reduced tumor cell viability was not toxic to normal cells. [2]
In prostate cancer studies, Raddeanin A did not inhibit the growth of AR-null PC-3 and DU145 cells, suggesting selectivity for AR-positive cells. [3]

[1]. Raddeanin A, a natural triterpenoid saponin compound, exerts anticancer effect on human osteosarcoma via the ROS/JNK and NF-κB signal pathway. Toxicol Appl Pharmacol. 2018 Aug 15;353:87-101.

[2]. Raddeanin A promotes apoptosis and ameliorates 5-fluorouracil resistance in cholangiocarcinoma cells. 3.411World J Gastroenterol. 2019 Jul 14;25(26):3380-3391.

[3]. Raddeanin A down-regulates androgen receptor and its splice variants in prostate cancer. J Cell Mol Med. 2019 May;23(5):3656-3664.

[4]. Raddeanin A Induces Apoptosis and Cycle Arrest in Human HCT116 Cells through PI3K/AKT Pathway Regulation In Vitro and In Vivo. Evid Based Complement Alternat Med. 2019 May 26;2019:7457105.

Raddeanin A is a triterpenoid compound.

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Raddeanin A induces apoptosis in osteosarcoma cells via the ROS/JNK signaling pathway and suppresses metastasis by inhibiting the NF-κB pathway and down-regulating MMP-2 and MMP-9. [1]
Raddeanin A acts as an anti-cancer agent and enhancer of 5-fluorouracil in bile duct cancer cells via regulating multiple cell cycle and apoptosis-related proteins (Wee1, COX-2, Bax, Bcl-2, cyclin D1, cyclin E). [2]
Raddeanin A down-regulates androgen receptor and its splice variants in prostate cancer by inducing proteasome-mediated degradation and inhibiting AR gene transcription. It enhances docetaxel efficacy synergistically. [3]
Raddeanin A induces apoptosis and G0/G1 cycle arrest in human colorectal cancer HCT116 cells through regulation of the PI3K/AKT signaling pathway. [4]

C47H76O16

897.0968

896.513

89412-79-3

174742

White to off-white solid

1.4±0.1 g/cm3

967.2±65.0 °C at 760 mmHg

276.2±27.8 °C

0.0±0.6 mmHg at 25°C

1.610

10.11

9

16

8

63

1720

22

C[C@]12[C@@]3(CC[C@H]4C(C)(C)[C@@H](O[C@@H]5OC[C@H](O)[C@H](O)[C@H]5O[C@@H]5O[C@H](CO)[C@@H](O)[C@H](O)[C@H]5O[C@@H]5O[C@@H](C)[C@H](O)[C@@H](O)[C@H]5O)CC[C@@]4([C@H]3CC=C1[C@@H]1CC(C)(C)CC[C@@]1(CC2)C(=O)O)C)C

VQQGPFFHGWNIGX-PPCHTBMASA-N

InChI=1S/C47H76O16/c1-22-30(50)33(53)35(55)38(59-22)62-37-34(54)32(52)26(20-48)60-40(37)63-36-31(51)25(49)21-58-39(36)61-29-12-13-44(6)27(43(29,4)5)11-14-46(8)28(44)10-9-23-24-19-42(2,3)15-17-47(24,41(56)57)18-16-45(23,46)7/h9,22,24-40,48-55H,10-21H2,1-8H3,(H,56,57)/t22-,24-,25-,26+,27-,28+,29-,30-,31-,32+,33+,34-,35+,36+,37+,38-,39-,40-,44-,45+,46+,47-/m0/s1

(4aS,6aR,6aS,6bR,8aR,10S,12aR,14bS)-10-[(2S,3R,4S,5S)-3-[(2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxyoxan-2-yl]oxy-4,5-dihydroxyoxan-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-carboxylic acid

2934.99.9001

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.

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

DMSO : ~50 mg/mL (~55.74 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (2.79 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 (2.79 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 (2.79 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.)