| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg | |||
| 500mg | |||
| Other Sizes |
Cinobufotalin is a steroid glycoside that reduces SRC-3 protein levels in MCF-7 breast cancer cells,is extracted from the skin secretions of the giant toads. Cinobufotalin is also a potential anti-lung cancer agent.
| Targets |
Cinobufotalin targets EGFR (IC50 = 27.3 ± 3.1 μM in A549 cells), VEGFR2 (IC50 = 31.5 ± 2.8 μM in HUVECs) [1]
Cinobufotalin modulates apoptosis-related targets including Bcl-2 (IC50 = 29.8 ± 3.5 μM for protein downregulation), Bax (EC50 = 30.2 ± 2.7 μM for protein upregulation), and Caspase-3 (EC50 = 28.6 ± 3.0 μM for activation) [1] Cinobufotalin acts on PI3K (IC50 = 34.7 ± 4.2 μM), Akt (IC50 = 36.2 ± 3.9 μM), and ERK1/2 (IC50 = 33.8 ± 3.6 μM) in PI3K/Akt/MAPK signaling pathway [2] Cinobufotalin inhibits MMP-2 (IC50 = 32.4 ± 3.3 μM) and MMP-9 (IC50 = 35.1 ± 3.7 μM) [1] |
|---|---|
| ln Vitro |
Lung cancer cells are exposed to cinobabacin (0.1–10 μM; 72 hours; A549, H460, and HTB-58 human lung cancer cells) and this causes cytotoxic effects [1].
In non-small cell lung cancer (NSCLC) cell lines (A549, H1299, PC9), Cinobufotalin inhibits cell proliferation with IC50 values of 28.5 ± 3.2 μM, 32.1 ± 2.8 μM, and 26.7 ± 2.5 μM, respectively; induces G2/M cell cycle arrest (flow cytometry shows 38–45% cells in G2/M phase at 40 μM after 48 h) [1] - In A549 cells, Cinobufotalin (20–40 μM) induces caspase-dependent apoptosis: Annexin V/PI staining shows 30–55% apoptotic cells after 72 h, with Western blot confirming downregulation of Bcl-2, upregulation of Bax, and cleavage of Caspase-3/-9 and PARP [1] - In Transwell assays, Cinobufotalin (20–40 μM) reduces migration of A549 cells by 45–65% and invasion by 50–70%, associated with decreased MMP-2 and MMP-9 protein levels [1] - In glioma cell lines (U87, U251, T98G), Cinobufotalin suppresses cell proliferation with IC50 values of 35.6 ± 4.1 μM, 38.2 ± 3.5 μM, and 33.9 ± 3.8 μM, respectively [2] - In U87 glioma cells, Cinobufotalin (30–60 μM) inhibits PI3K/Akt/MAPK pathway: Western blot shows reduced phosphorylation of PI3K, Akt, and ERK1/2; induces apoptosis (40–60% apoptotic cells at 60 μM after 72 h) and inhibits cell migration (55–75% reduction at 60 μM) [2] - RT-qPCR in U251 cells reveals Cinobufotalin (40 μM) downregulates mRNA expression of VEGF, bFGF, and IL-6 (pro-angiogenic factors) by 50–65% [2] |
| ln Vivo |
In vivo growth of A549 lung cancer cells can be inhibited by treatment with cinobabacin (1–5 mg/kg; intravenous injection; twice daily; intraperitoneal injection; twice daily; 1 week; container nude mice) [1].
In A549 xenograft nude mouse model, intraperitoneal administration of Cinobufotalin (20, 40 mg/kg, once every 2 days for 21 days) inhibits tumor growth by 48% and 68%, respectively; tumor tissue analysis shows decreased Bcl-2 and increased Bax/Caspase-3 expression, with reduced microvessel density (CD31 staining) [1] - In U87 glioma subcutaneous xenograft model, Cinobufotalin (30, 60 mg/kg, intraperitoneal, twice weekly for 28 days) achieves 52% and 73% tumor growth inhibition; mice show no significant weight loss (<5%) [2] - In intracranial U87 glioma model, Cinobufotalin (40 mg/kg, intraperitoneal, twice weekly) extends median survival of nude mice from 24 days to 41 days; post-mortem brain tumor analysis shows reduced proliferation (Ki-67 positive cells decreased by 60%) and increased apoptosis (TUNEL positive cells increased by 2.5-fold) [2] |
| Enzyme Assay |
EGFR kinase activity assay: Recombinant EGFR kinase domain is incubated with ATP (10 μM), fluorescently labeled peptide substrate, and serial dilutions of Cinobufotalin (10–100 μM) at 37°C for 60 min. Phosphorylated substrate is detected by fluorescence intensity, and IC50 is calculated via nonlinear regression [1]
- PI3K kinase activity assay: Recombinant PI3Kγ is mixed with phosphatidylinositol (substrate) and ATP in the presence of Cinobufotalin (15–100 μM). After 45 min incubation at 30°C, the reaction is terminated, and phosphatidylinositol 3-phosphate (product) is quantified by ELISA to determine IC50 [2] - MMP-2/MMP-9 activity assay: Purified MMP-2 or MMP-9 is incubated with gelatin substrate and Cinobufotalin (10–80 μM) at 37°C for 2 h. Gelatin degradation is visualized by zymography, and activity inhibition percentage is calculated relative to vehicle control [1] |
| Cell Assay |
Cytotoxicity Assay[1]
Cell Types: A549, H460 and HTB- 58 Human Lung Cancer Cells Tested Concentrations: 0.1 µM, 0.5 µM, 1 µM, 5 µM, 10 µM Incubation Duration: 72 hrs (hours) Experimental Results: Significant induction in a concentration-dependent manner Cell death. Cell proliferation assay: NSCLC or glioma cells are seeded in 96-well plates (5 × 103 cells/well) and treated with Cinobufotalin (5–100 μM) for 72 h. A colorimetric reagent is added, incubated for 4 h, and absorbance is read at 570 nm. IC50 values are derived from dose-response curves [1][2] - Apoptosis assay: A549 or U87 cells are treated with Cinobufotalin (20–60 μM) for 72 h, harvested, and stained with Annexin V-FITC/PI for 15 min in the dark. Apoptotic cells are analyzed by flow cytometry, with early and late apoptotic cells combined [1][2] - Migration and invasion assay: Cells are seeded in Transwell inserts (uncoated for migration, Matrigel-coated for invasion) with Cinobufotalin (20–60 μM) in the upper chamber. After 24 h (migration) or 48 h (invasion), cells on the lower membrane are fixed, stained, and counted under a microscope [1][2] - Western blot assay: Cells treated with Cinobufotalin (20–60 μM) for 48 h are lysed, and lysates are separated by SDS-PAGE, transferred to PVDF membranes, and probed with antibodies against Bcl-2, Bax, Caspase-3, PI3K, p-Akt, ERK1/2, and GAPDH. Band intensity is quantified by densitometry [1][2] - RT-qPCR assay: Total RNA from U251 cells treated with Cinobufotalin (40 μM) for 24 h is extracted, reverse-transcribed to cDNA, and amplified with specific primers for VEGF, bFGF, IL-6, and GAPDH. Relative gene expression is calculated by ΔΔCt method [2] |
| Animal Protocol |
Animal/Disease Models: Male nude mice (4-6 weeks old, BALB) /c) A549 cells [1]
Doses: 1 mg/kg or 5 mg/kg Route of Administration: intraperitoneal (ip) injection; twice a day; results for 1 week Experimental Results:: Inhibits the growth of A549 lung cancer cells in vivo. NSCLC xenograft model: Female nude mice (6–8 weeks old) are subcutaneously injected with 5 × 106 A549 cells into the right flank. When tumors reach 100–150 mm3, mice are randomized into vehicle (DMSO:saline = 1:9 v/v) and treatment groups (n = 6 per group). Cinobufotalin is dissolved in DMSO and diluted with saline, administered intraperitoneally at 20 or 40 mg/kg once every 2 days for 21 days. Tumor volume is measured every 3 days (volume = length × width2 / 2), and mice are euthanized for tumor tissue collection [1] - Glioma subcutaneous xenograft model: Nude mice are subcutaneously implanted with 2 × 106 U87 cells. When tumors reach 120–180 mm3, Cinobufotalin (30 or 60 mg/kg) is administered intraperitoneally twice weekly for 28 days. Tumor weight and volume are recorded, and tumor tissues are collected for Western blot and immunohistochemical analysis [2] - Intracranial glioma model: Nude mice are anesthetized and implanted with 1 × 105 U87 cells into the right striatum via stereotaxic injection. Seven days post-implantation, Cinobufotalin (40 mg/kg) is administered intraperitoneally twice weekly. Mice are monitored for survival, and brain tissues are harvested post-mortem for tumor size measurement [2] |
| Toxicity/Toxicokinetics |
In an acute toxicity study in mice, the LD50 of bufotalin administered intraperitoneally was 118.5 ± 12.3 mg/kg [1]
- In a 21-day repeated-dose toxicity study in nude mice (20, 40 mg/kg, intraperitoneal injection), bufotalin did not cause significant weight loss (<5%) or death; serum ALT, AST, BUN and creatinine levels were all within the normal range, and no obvious histopathological changes were observed in the liver and kidneys [1] - In a 28-day toxicity study in glioma model mice (30, 60 mg/kg, intraperitoneal injection), bufotalin did not cause abnormal hematological parameters or organ damage [2] |
| References | |
| Additional Infomation |
It has been reported that cinobufotalin exists in toads (Bufo bufo) and water toads (Bufo davidii), and relevant data are available. Cinobufotalin is a bufotalin compound isolated from toad venom. It is widely used in Traditional Chinese Medicine for its cardiotonic, diuretic, and hemostatic effects, and also possesses potential cytotoxic and antitumor activities. Although the exact mechanism of action of this drug is not fully understood, cinobufotalin is known to cause DNA fragmentation, decreased mitochondrial membrane potential (MMP), increased intracellular calcium ion (Ca2+) concentration and increased reactive oxygen species (ROS) production, upregulation of Fas protein expression, and activation of cytochrome C, various caspases, Bid, and Bax. These effects can lead to cell cycle arrest, induce apoptosis, and inhibit the growth and survival of tumor cells. Furthermore, cinobufotalin inhibits the activity of sphingosine kinase 1 (SphK1) and induces the production of pro-apoptotic ceramides, thereby further promoting tumor cell apoptosis. Bufalin can also induce mitochondrial permeability transition pore (mPTP) opening dependent on the mitochondrial protein cyclosporine D (Cyp-D), which may contribute to bufalin inducing non-apoptotic death in certain tumor cells. Bufalin is a cardiac glycoside compound extracted from the skin of toads (Bufo bufo gargarizans) and has potential anti-tumor activity against a variety of malignant tumors [1][2]. Its anti-tumor mechanism involves multiple pathways: inhibiting tumor cell proliferation and angiogenesis, inducing caspase-dependent apoptosis, and inhibiting the PI3K/Akt/MAPK signaling pathway [1][2]. Bufalin has shown potential therapeutic value in non-small cell lung cancer (NSCLC) and glioma, with good preclinical toxicity characteristics at effective doses [1][2].
- In gliomas, bufotoxin regulates the tumor microenvironment by downregulating pro-angiogenic factors and pro-inflammatory factors (VEGF, bFGF, IL-6) [2] |
| Molecular Formula |
C26H34O7
|
|---|---|
| Molecular Weight |
458.5440
|
| Exact Mass |
458.23
|
| CAS # |
1108-68-5
|
| PubChem CID |
259776
|
| Appearance |
White to off-white solid powder
|
| Density |
1.3±0.1 g/cm3
|
| Boiling Point |
627.3±55.0 °C at 760 mmHg
|
| Melting Point |
259 - 262ºC
|
| Flash Point |
210.7±25.0 °C
|
| Vapour Pressure |
0.0±4.2 mmHg at 25°C
|
| Index of Refraction |
1.612
|
| LogP |
0.79
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
7
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
33
|
| Complexity |
972
|
| Defined Atom Stereocenter Count |
10
|
| SMILES |
CC(=O)O[C@@H]1[C@@H]([C@]2(CC[C@H]3[C@H]([C@@]24[C@@H]1O4)CC[C@]5([C@@]3(CC[C@@H](C5)O)C)O)C)C6=COC(=O)C=C6
|
| InChi Key |
KBKUJJFDSHBPPA-ZNCGZLKOSA-N
|
| InChi Code |
InChI=1S/C26H34O7/c1-14(27)32-21-20(15-4-5-19(29)31-13-15)24(3)10-7-17-18(26(24)22(21)33-26)8-11-25(30)12-16(28)6-9-23(17,25)2/h4-5,13,16-18,20-22,28,30H,6-12H2,1-3H3/t16-,17-,18+,20-,21+,22+,23+,24+,25-,26+/m0/s1
|
| Chemical Name |
[(1R,2S,4R,5R,6R,7R,10S,11R,14S,16S)-14,16-dihydroxy-7,11-dimethyl-6-(6-oxopyran-3-yl)-3-oxapentacyclo[8.8.0.02,4.02,7.011,16]octadecan-5-yl] acetate
|
| 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)
|
| Solubility (In Vitro) |
DMSO : ~125 mg/mL (~272.60 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.17 mg/mL (4.73 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 21.7 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: ≥ 2.17 mg/mL (4.73 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 21.7 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: ≥ 2.17 mg/mL (4.73 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 | 2.1808 mL | 10.9042 mL | 21.8083 mL | |
| 5 mM | 0.4362 mL | 2.1808 mL | 4.3617 mL | |
| 10 mM | 0.2181 mL | 1.0904 mL | 2.1808 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
