Crebanine targets multiple signaling pathways and proteins. It acts as an antagonist of the α7-nicotinic acetylcholine receptor with an IC50 of 19.1 μM. It inhibits the AKT/FoxO3a, NF-κB, and MAPK signaling pathways. It also targets Akt, apoptosis-related proteins, NF-κB, reactive oxygen species, p38 MAPK, ERK, interleukin-related proteins, TNF receptor, NO synthase, and nAChR.

Crebanine suppresses the proliferation, migration, and invasion of cancer cells, triggers reactive oxygen species (ROS) burst, and promotes apoptosis. It inhibits the AKT/FoxO3a, NF-κB, and MAPK signaling pathways. Crebanine attenuates NOX2 hyperactivation and exhibits antioxidant properties by reducing reactive oxygen species and peroxidation in microglia cells. It inhibits voltage-dependent Na+ current in guinea-pig ventricular myocytes.

Crebanine inhibits the proliferation of human leukemic cells (HL-60, U937 and K562), human fibrosarcoma cells (HT1080) and cervix cancer cell lines (KB-3-1 and KB-V1). It sensitizes TNF-α induced apoptosis and suppresses human lung adenocarcinoma. Crebanine ameliorates ischemia-reperfusion brain damage in middle cerebral artery occlusion and reperfusion (MCAO/R) rats. It significantly improves scopolamine-induced cognitive deficits in ICR mice.

BGT226 maleate (NVP-BGT226 maleate) is aPI3K(withIC50s of 4 nM, 63 nM and 38 nM forPI3Kα,PI3KβandPI3Kγ) /mTORdual inhibitor which displays potent growth-inhibitory activity against human head and neck cancer cells.
Crebanine is evaluated in cell-free receptor binding assays to determine its affinity for the α7-nicotinic acetylcholine receptor. Radioligand binding displacement assays are performed using membrane preparations expressing the receptor. Competition binding experiments are conducted with increasing concentrations of Crebanine and a fixed concentration of a labeled reference ligand. IC50 values are determined to quantify the potency of receptor antagonism.

NCI-H929, U266, RPMI-8226 and OPM2 MM cells are seeded in 96-well plates at a concentration of 1.5 × 104cells/well in RPMI medium supplemented with 10% fetal bovine serum with or without NVP-BGT226 that is to be tested. After 36 hours, BrdU-labelling solution is added (final concentration: 10 μM), and cells are cultured for another 12 hours in a humidified atmosphere (37 °C/5% CO2). Then, the plates are centrifuged (10 min, 300 g), and the supernatants are discarded. The plates are dried at 60 °C for 2 hours. After fixation with ethanol/HCl for 30 min at -20 °C, the DNA is partially digested by nuclease treatment for 30 min at 37 °C. The cells are washed three times with medium and incubated with anti-BrdU-POD labelling solution for 30 min at 37 °C. The anti-POD solution is removed and the cells are washed three times with washing buffer. The ABTS substrate solution is added, and absorbance is measured in a microplate reader at 405 nm with a reference wave length of 490 nm.The anti-proliferative and pro-apoptotic effects of NVP-BGT226 are independent of bcr-abl status. The activation of the AKT/mTOR signal cascade is suppressed by NVP-BGT226 in a concentration- and time-dependent manner. Flow cytometric analysis exhibits an accumulation of cells in the G(0)-G(1) phase with concomitant loss in the S-phase. NVP-BGT226 displays potent growth-inhibitory activity against all tested cell lines including SCC4, TU183 and KB cell lines with the IC50 ranging from 7.4 to 30.1 nM. Notably, both Detroit 562 and HONE-1 cells, which express PIK3CA mutation H1047R, are still sensitive to the growth-inhibitory effect of NVP-BGT226 treatment. In addition, the sensitivity to NVP-BGT226 between HONE-1 cells and its cisplatin-resistant variant is almost identical. Results of the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay and the analysis of caspase 3/7 and PARP indicates that NVP-BGT226 induces cancer cell death through an apoptosis-independent pathway. NVP-BGT226 induces autophagy as indicated by the aggregation and upregulation of the microtubule-associated protein light chain 3B-II, and p62 degradation. Gene silencing of Beclin1 or cotreatment of the autophagosome inhibitor, 3-methyladenine, inhibits the NVP-BGT226-induced autophagy and leads to the retrieval of colony survival.
Crebanine is assessed in cell-based assays using various cancer cell lines, including HL-60, U937, K562, HT1080, KB-3-1, KB-V1, A549 lung, MDA-MB-231 and MCF-7 breast, and SKOV3 and Caov-3 ovarian cells. Cells are treated with Crebanine, and cell proliferation, migration, invasion, and apoptosis are measured. ROS production and signaling pathway modulation are assessed using Western blot analysis.

Crebanine is administered in animal models to evaluate its anticancer, anti-inflammatory, and cognitive-enhancing effects. It ameliorates ischemia-reperfusion brain damage in MCAO/R rats. It significantly improves scopolamine-induced cognitive deficits in ICR mice. Efficacy is assessed by measuring tumor growth, inflammatory markers, and cognitive performance.

Crebanine has a molecular weight of 339.39 and a molecular formula of C20H21NO4. It has a CAS number of 25127-29-1. The compound is soluble in DMSO at 100 mg/mL. In vivo, it can be formulated as a clear solution using 10% DMSO, 40% PEG300, 5% Tween-80, and 45% saline. It should be stored at 4°C and protected from light.

No detailed toxicity data is available for Crebanine. The compound is for research use only and is not intended for human therapeutic use. It has shown high inhibitory activity against gram-positive animal pathogenic bacteria.

[1]. Induction of G1 arrest and apoptosis in human cancer cells by crebanine, an alkaloid from Stephania venosa. Chem Pharm Bull (Tokyo). 2012;60(10):1283-9.

[2]. Antiinflammatory Activities of Crebanine by Inhibition of NF-κB and AP-1 Activation through Suppressing MAPKs and Akt Signaling in LPS-Induced RAW264.7 Macrophages. Biol Pharm Bull. 2016;39(1):54-61.

Klebanon is an isoquinoline alkaloid. (12R)-15,16-dimethoxy-11-methyl-3,5-dioxa-11-azapentacyclo[10.7.1.02,6.08,20.014,19]eicos-1(20),2(6),7,14(19),15,17-hexene has been reported in Stephania cephalantha, Stephania venosa, and other organisms with available data. See also: Klebanon (note moved to).
Crebanine is an aporphine alkaloid. It has been studied for its ability to inhibit acetylcholinesterase, making it of interest in research related to neurodegenerative diseases such as Alzheimer's. It can be used for the study of hepatocellular carcinoma (HCC). The compound is not approved for clinical use.

C20H21NO4

339.391

339.147

C, 70.78; H, 6.24; N, 4.13; O, 18.86

25127-29-1

25127-29-1

333544

Off-white to light yellow solid powder

1.3±0.1 g/cm3

488.2±45.0 °C at 760 mmHg

145.4±25.9 °C

0.0±1.2 mmHg at 25°C

1.616

3.98

0

5

2

25

502

1

O1C([H])([H])OC2C([H])=C3C([H])([H])C([H])([H])N(C([H])([H])[H])C4([H])C([H])([H])C5C(=C(C([H])=C([H])C=5C(C1=2)=C43)OC([H])([H])[H])OC([H])([H])[H]

UVDQDNQWGQFIAO-UHFFFAOYSA-N

InChI=1S/C20H21NO4/c1-21-7-6-11-8-16-20(25-10-24-16)18-12-4-5-15(22-2)19(23-3)13(12)9-14(21)17(11)18/h4-5,8,14H,6-7,9-10H2,1-3H3

15,16-dimethoxy-11-methyl-3,5-dioxa-11-azapentacyclo[10.7.1.02,6.08,20.014,19]icosa-1(20),2(6),7,14(19),15,17-hexaene

Crebanin; Crebanine

2934.99.03.00

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: 68~100 mg/mL (200.4~294.7 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.37 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 (7.37 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 (7.37 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.)