| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| 10mg |
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| 100mg | |||
| Other Sizes |
| Targets |
- Gamabufotalin targets the IKKβ/NF-κB signaling pathway [1]
- Gamabufotalin targets the STAT3/Mcl-1 signaling pathway [2] - Gamabufotalin targets ATP1A3 (specifically the Thr794 residue) and the AQP4 pathway [3] - Gamabufotalin targets the VEGFR-2 signaling pathway (IC50 for VEGFR-2 kinase inhibition: ~1.5 μM) [4] |
|---|---|
| ln Vitro |
- For lung cancer cells (A549, H460): Gamabufotalin (0.5–5 μM) inhibited cell proliferation in a dose-dependent manner (IC50: ~2.5 μM, MTT assay). It downregulated COX-2 protein and mRNA expression (western blot, RT-PCR) by suppressing IKKβ phosphorylation and NF-κB nuclear translocation (immunofluorescence); 5 μM Gamabufotalin reduced COX-2 mRNA by ~70% vs. control [1]
- For breast cancer cells (MDA-MB-231, MCF-7): Gamabufotalin (1–4 μM) sensitized cells to TRAIL-induced apoptosis. At 3 μM, it increased TRAIL-induced apoptotic rate by ~45% (Annexin V-FITC/PI staining) by inhibiting STAT3 phosphorylation and downregulating Mcl-1 (western blot); ≤3 μM Gamabufotalin alone had no significant effect on cell viability [2] - For glioblastoma cells (U87, U251): Gamabufotalin (0.5–3 μM) enhanced temozolomide (TMZ) sensitivity. The IC50 of TMZ decreased from ~80 μM to ~25 μM when combined with 1.8 μM Gamabufotalin (MTT assay). It reduced ATP1A3 expression and activated the AQP4 pathway (western blot), promoting TMZ-induced DNA damage [3] - For human umbilical vein endothelial cells (HUVEC): Gamabufotalin (0.5–4 μM) inhibited VEGF-induced angiogenesis. At 2 μM, it reduced tube formation by ~60% (tube formation assay) and cell migration by ~55% (transwell assay) by suppressing VEGFR-2 phosphorylation and downstream AKT/ERK signaling (western blot); IC50 for HUVEC proliferation inhibition: ~2 μM [4] |
| ln Vivo |
- For nude mouse lung cancer xenografts (A549): Gamabufotalin (5, 10 mg/kg, intraperitoneal injection, once every 2 days for 21 days) reduced tumor volume by ~65% and weight by ~60% vs. control (calipers, weighing). Tumor tissues showed downregulated p-IKKβ, nuclear NF-κB, and COX-2 (immunohistochemistry, western blot) [1]
- For nude mouse glioblastoma xenografts (U87): Mice were treated with Gamabufotalin (4, 8 mg/kg, intraperitoneal injection, once every 2 days) + TMZ (20 mg/kg, oral, daily) for 28 days. The high-dose combination reduced tumor volume by ~75% vs. TMZ alone; tumor tissues had decreased ATP1A3 and increased AQP4 (western blot) [3] - For chicken chorioallantoic membrane (CAM) and nude mouse breast cancer xenografts (MDA-MB-231): In CAM, Gamabufotalin (2, 4 mg/kg) reduced blood vessel density by ~40%, ~60% (microscopy). In xenografts, 4 mg/kg Gamabufotalin (intraperitoneal injection, once every 2 days for 21 days) reduced tumor microvessel density by ~55% (CD31 immunohistochemistry) vs. control [4] |
| Enzyme Assay |
- IKKβ kinase activity assay: Recombinant human IKKβ was incubated with ATP (10 μM), IKKβ-specific substrate peptide, and Gamabufotalin (0.5–5 μM) at 37°C for 45 minutes. The reaction was terminated with stop buffer, and phosphorylated substrate was detected via ELISA; Gamabufotalin inhibited IKKβ activity with an IC50 of ~1.8 μM [1]
- VEGFR-2 kinase activity assay: Recombinant human VEGFR-2 (kinase domain) was incubated with ATP (5 μM), fluorescent substrate, and Gamabufotalin (0.5–4 μM) at 37°C for 60 minutes. Fluorescence intensity of phosphorylated substrate was measured (excitation 485 nm, emission 535 nm); Gamabufotalin had an IC50 of ~1.5 μM for VEGFR-2 [4] |
| Cell Assay |
- Lung cancer cell (A549) assay: Cells (5×10³/well, 96-well plate) were treated with Gamabufotalin (0.5–5 μM) for 72 hours; viability was measured via MTT assay. Cells (6-well plate) were treated with 5 μM Gamabufotalin for 24 hours; p-IKKβ, NF-κB, COX-2 were analyzed via western blot, and NF-κB nuclear translocation via immunofluorescence [1]
- Breast cancer cell (MDA-MB-231) apoptosis assay: Cells (6-well plate) were pre-treated with Gamabufotalin (1–4 μM) for 12 hours, then TRAIL (10 ng/mL) for 24 hours. Apoptosis was detected via Annexin V-FITC/PI staining + flow cytometry; p-STAT3, Mcl-1 via western blot [2] - Glioblastoma cell (U87) sensitivity assay: Cells (96-well plate) were treated with Gamabufotalin (0.5–3 μM) + TMZ (10–100 μM) for 72 hours; IC50 was calculated via MTT assay. Cells treated with 1.8 μM Gamabufotalin for 24 hours were lysed; ATP1A3, AQP4 were analyzed via western blot [3] - HUVEC angiogenesis assay: HUVECs on Matrigel-coated 96-well plates were treated with Gamabufotalin (0.5–4 μM) + VEGF (20 ng/mL) for 6 hours; tube formation was counted via microscopy. HUVECs treated with Gamabufotalin for 12 hours were seeded in transwell upper chambers (8 μm pores), VEGF in lower chambers; migrated cells were stained and counted [4] |
| Animal Protocol |
- Lung cancer xenograft (A549) protocol: BALB/c nude mice (4–6 weeks) were subcutaneously injected with 1×10⁷ A549 cells. When tumors reached ~100 mm³, mice (n=6/group) received Gamabufotalin (5, 10 mg/kg) via intraperitoneal injection once every 2 days for 21 days. Tumor volume (length×width²/2) was measured every 3 days; mice were euthanized, tumors weighed, and tissues collected for analysis [1]
- Glioblastoma xenograft (U87) protocol: Nude mice were subcutaneously injected with 2×10⁶ U87 cells. When tumors reached ~150 mm³, mice (n=6/group) received Gamabufotalin (4, 8 mg/kg, intraperitoneal injection, once every 2 days) + TMZ (20 mg/kg, oral, daily) for 28 days. Tumor volume was measured, and tissues collected for western blot [3] - Breast cancer xenograft (MDA-MB-231) protocol: Mice were subcutaneously injected with 1×10⁷ MDA-MB-231 cells. When tumors reached ~100 mm³, mice received 4 mg/kg Gamabufotalin via intraperitoneal injection once every 2 days for 21 days. Tumors were excised, and microvessel density detected via CD31 immunohistochemistry [4] |
| Toxicity/Toxicokinetics |
In vitro toxicity: ≤5 μM Gamabufotalin showed no significant cytotoxicity to normal human lung fibroblasts (MRC-5) or HUVEC cells (MTT assay, cell viability >80%) [1,4]
- In vivo toxicity: In nude mouse experiments (21–28 days), ≤10 mg/kg Gamabufotalin did not cause significant weight loss (<5% vs. control group) or abnormal pathological changes in the heart, liver, or kidneys (hematoxylin-eosin staining) [1,3,4] |
| References |
|
| Additional Infomation |
γ-Bufotalin is a steroidal lactone. Functionally, it is associated with bufotalin. Gamabufotalin has been reported in toads (Bufo bufo), green toads (Bufotes viridis), and other organisms with relevant data.
- Gamabufotoxin is a bufodiene lactone compound isolated from toad venom (Venenum Bufonis)[1] - Gamabufotoxin enhances TRAIL-induced apoptosis in breast cancer cells by blocking the STAT3/Mcl-1 anti-apoptotic pathway, thereby overcoming TRAIL resistance[2] - Gamabufotoxin promotes the sensitivity of glioblastoma to temozolomide through a negative feedback loop: targeting ATP1A3 Thr794 → reducing ATP1A3 → activating AQP4 → enhancing temozolomide-induced DNA damage[3] - Gamabufotoxin has anti-angiogenic effects by inhibiting the VEGFR-2/AKT/ERK signaling pathway, supporting its potential in the treatment of angiogenesis-dependent tumors[4] |
| Molecular Formula |
C24H34O5
|
|---|---|
| Molecular Weight |
402.5238
|
| Exact Mass |
402.24
|
| CAS # |
465-11-2
|
| PubChem CID |
259803
|
| Appearance |
White to off-white solid powder
|
| Density |
1.3±0.1 g/cm3
|
| Boiling Point |
595.8±50.0 °C at 760 mmHg
|
| Flash Point |
203.4±23.6 °C
|
| Vapour Pressure |
0.0±3.8 mmHg at 25°C
|
| Index of Refraction |
1.612
|
| LogP |
1.99
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
1
|
| Heavy Atom Count |
29
|
| Complexity |
773
|
| Defined Atom Stereocenter Count |
9
|
| SMILES |
C[C@]12CC[C@@H](C[C@H]1CC[C@@H]3[C@@H]2[C@@H](C[C@]4([C@@]3(CC[C@@H]4C5=COC(=O)C=C5)O)C)O)O
|
| InChi Key |
FMTLOAVOGWSPEF-KJRPADTMSA-N
|
| InChi Code |
InChI=1S/C24H34O5/c1-22-9-7-16(25)11-15(22)4-5-18-21(22)19(26)12-23(2)17(8-10-24(18,23)28)14-3-6-20(27)29-13-14/h3,6,13,15-19,21,25-26,28H,4-5,7-12H2,1-2H3/t15-,16+,17-,18-,19-,21-,22+,23-,24+/m1/s1
|
| Chemical Name |
5-[(3S,5R,8R,9S,10S,11R,13R,14S,17R)-3,11,14-trihydroxy-10,13-dimethyl-1,2,3,4,5,6,7,8,9,11,12,15,16,17-tetradecahydrocyclopenta[a]phenanthren-17-yl]pyran-2-one
|
| 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 |
| 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 : ~50 mg/mL (~124.22 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.21 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (6.21 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (6.21 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 | 2.4843 mL | 12.4217 mL | 24.8435 mL | |
| 5 mM | 0.4969 mL | 2.4843 mL | 4.9687 mL | |
| 10 mM | 0.2484 mL | 1.2422 mL | 2.4843 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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