- TRAF6 (TNF receptor-associated factor 6): Tabersonine suppresses K63-linked polyubiquitination of TRAF6. [1]
- NLRP3 (NACHT, LRR, and PYD domains-containing protein 3): IC50 = 0.71 μM (for inhibiting IL-1β production in BMDMs). Tabersonine directly binds to the NACHT domain of NLRP3. [2]
- PI3K/Akt pathway: Tabersonine inhibits Akt phosphorylation (Ser473). [3]

- ALI model (Macrophages): Tabersonine (1, 3, 10 μM) had no cytotoxicity in BMDMs up to 10 μM. It reduced LPS-induced iNOS protein level and NO release in a dose-dependent manner. It also reduced the mRNA and protein levels of TNF-α, IL-6, and IL-1β in LPS-stimulated BMDMs. Tabersonine suppressed LPS-induced phosphorylation of p65 NF-κB, IκB-α degradation, and NF-κB transcriptional activity. It also suppressed the phosphorylation of p38 MAPK and its downstream kinase MK2, with weak effects on ERK and JNK. Tabersonine (10 μM) significantly reduced K63-linked polyubiquitination of TRAF6 but had no effect on K48-linked ubiquitination. [1]
- NLRP3 inflammasome (Macrophages): Tabersonine potently inhibited LPS+ATP-induced IL-1β production in BMDMs (IC50 = 0.71 μM) and THP-1 cells. It suppressed caspase-1 (p20) cleavage and IL-1β secretion dose-dependently (1, 5, 10 μM). It had no effect on TNF-α or IL-6 levels, nor on NLRP3 or pro-IL-1β expression, indicating an NF-κB-independent mechanism. Tabersonine inhibited LDH release, pyroptosis (GSDMD-NT), and ASC speck formation and oligomerization. It directly bound to NLRP3, protecting it from pronase degradation in DARTS assay, specifically binding to the NACHT domain. Tabersonine inhibited NLRP3 self-oligomerization and its ATPase activity. It blocked the NLRP3-ASC interaction but not the NLRP3-NEK7 interaction. [2]
- Hepatocellular carcinoma (HCC): Tabersonine inhibited the viability of SMMC7721 (IC50 = 7.89 ± 1.2 μM), Bel7402 (IC50 = 5.07 ± 1.4 μM), and HepG2 (IC50 = 12.39 ± 0.7 μM) cells. It significantly inhibited colony formation in all three cell lines (6-30 μM). Tabersonine induced apoptosis in HepG2 cells, as shown by Hoechst 33258, AO/EB, and Annexin V-FITC/PI staining (apoptotic rate reached 27% at 30 μM). It increased cleaved Caspase-3 and cleaved PARP levels, reduced mitochondrial membrane potential (JC-1 staining), increased the Bax/Bcl-2 ratio, promoted cytochrome c release, and activated cleaved Caspase-9. Tabersonine downregulated p-Akt (Ser473) without affecting total Akt, and synergized with the PI3K inhibitor LY294002 to further inhibit p-Akt. It also increased Fas and FasL expression, decreased Caspase-8 and Bid levels, indicating activation of the death receptor pathway. [3]

- ALI model (LPS-induced): In C57BL/6 mice, intraperitoneal injection of tabersonine (10, 20, 40 mg/kg) prior to intratracheal LPS (5 mg/kg) significantly attenuated lung pathological injury, reduced total cells, neutrophils (Ly-6G+), and protein concentration in BALF, decreased MPO activity, and lowered TNF-α, IL-6, and IL-1β mRNA in lung tissue and protein levels in serum. [1]
- NLRP3-driven disease models: In an alum-induced peritonitis model, oral administration of tabersonine (10, 20, 40 mg/kg) in mice significantly decreased total cells, IL-1β levels, and monocyte/neutrophil counts in peritoneal fluid. In an LPS-induced ALI model, oral tabersonine (10 mg/kg) in WT mice, but not in Nlrp3KO mice, reduced lung pathological injury, wet/dry ratio, protein and total cells in BALF, and IL-1β (p20, mature) levels. In a sepsis model (E. coli infection), tabersonine (10 mg/kg) pretreatment significantly increased the survival rate of WT mice to 60% but had no effect on Nlrp3KO mice. [2]
- HCC xenograft model: In BALB/c nude mice bearing HepG2 xenografts, oral administration of tabersonine (25 or 50 mg/kg/day for 3 weeks) significantly inhibited tumor growth and tumor weight without affecting body weight. TUNEL staining and cleaved Caspase-3 immunofluorescence in tumor tissues confirmed increased apoptosis. [3]

- ATPase activity assay for NLRP3: Human NLRP3 immunoprecipitated from transfected HEK293T cells was incubated with different concentrations of tabersonine for 40 min. Ultra-pure ATP was then added to the reaction buffer and incubated at 37°C for 40 min. The amount of ATP converted to ADP was determined using a luminescent ADP detection kit. The data showed that tabersonine suppressed NLRP3 ATPase activity. [2]
- Drug Affinity Responsive Target Stability (DARTS): BMDMs were primed with LPS for 3h, or HEK-293T cells were harvested 24h after transfection. Total cell lysates were incubated with tabersonine overnight at 4°C. Pronase was added and incubated for 5 min at room temperature. The reaction was stopped by adding SDS loading buffer and heating. Samples were analyzed by immunoblotting. Tabersonine protected NLRP3, but not ASC, from pronase-driven hydrolysis. Domain mapping showed it specifically protected the NACHT domain. [2]

- Cell Viability (MTT): For BMDMs, cells were plated in 96-well plates, treated with tabersonine (0-10 μM) for 24h, then MTT was added for 4h. Absorbance was read at 490nm. Tabersonine had no cytotoxicity up to 10 μM. [1][2]
For HCC cells (HepG2, SMMC7721, Bel7402), cells were seeded in 96-well plates, treated with tabersonine (6-30 μM) for 24h, then MTT was added for 4h. Absorbance was read at 450nm. IC50 values were calculated. [3]
- Colony Formation Assay: HCC cells were seeded in 6-well plates, treated with tabersonine (6-30 μM). Medium was replaced every 3 days. After two weeks, colonies were fixed with 4% paraformaldehyde, stained with 0.2% crystal violet, and counted (diameter >75 μm). Tabersonine significantly inhibited colony formation. [3]
- Apoptosis Detection (Flow Cytometry): HepG2 cells were treated with tabersonine (6-30 μM) for 18h, then stained with Annexin V-FITC and PI. The apoptotic rate was analyzed by flow cytometry. [3]
For BMDMs, cells were stained with Annexin V-FITC and PI after LPS+ATP treatment with or without tabersonine. [1]
- Mitochondrial Membrane Potential (JC-1): HepG2 cells were treated with tabersonine (6-30 μM) for 24h, then stained with JC-1. Changes from red (high potential) to green (low potential) were observed by fluorescence microscopy. [3]
- Western Blotting: Cells or tissues were lysed, and proteins were separated by SDS-PAGE, transferred to membranes, and probed with specific antibodies for various targets (e.g., iNOS, p-p65, p-p38, IL-1β, cleaved Caspase-3, Bax, Bcl-2, p-Akt, etc.). [1][2][3]
- Quantitative Real-time PCR (qRT-PCR): Total RNA was extracted from BMDMs or lung tissues. cDNA was synthesized, and gene expression of TNF-α, IL-6, and IL-1β was measured using SYBR Green real-time PCR. [1]
- ELISA: Supernatants from cell cultures, BALF, peritoneal fluids, or serum were collected. The levels of IL-1β, TNF-α, and IL-6 were measured using commercial ELISA kits. [1][2]
- Immunofluorescence (IF): BMDMs were fixed, permeabilized, and stained with antibodies against caspase-1 or ASC. Nuclear counterstaining was performed with DAPI. Images were captured by confocal microscopy to visualize ASC specks or caspase-1 activation. [2]
For tumor tissues, frozen sections were stained with anti-cleaved Caspase-3 antibody. [3]
- Luciferase Reporter Assay: THP-1 cells stably expressing NF-κB luciferase reporter were treated with tabersonine (1, 3, 10 μM) for 1h, then stimulated with LPS (100 ng/mL) for 6h. Luciferase activity was measured. Tabersonine significantly reduced LPS-induced NF-κB activation. [1]

- ALI Model (Zhang et al. 2018): C57BL/6 mice were intraperitoneally (i.p.) injected with solvent control, dexamethasone (5 mg/kg), or tabersonine (10, 20, 40 mg/kg). One hour later, LPS (5 mg/kg) was administered intratracheally to induce ALI. Mice were euthanized 6h later for sample collection. Tabersonine was dissolved in a vehicle (H2O:ethanol:polyoxyethylene hydrogenated castor oil = 8:1:1). [1]
- Peritonitis Model (Xu et al. 2023): 10-week-old male C57BL/6 mice were treated via gavage with tabersonine (10, 20, or 40 mg/kg) dissolved in 0.5% carboxymethylcellulose sodium (CMC-Na) or vehicle. This was followed by intraperitoneal injection of Alum (1 mg per mouse). After 6h, mice were sacrificed, and peritoneal lavage was performed. [2]
- ALI Model (Xu et al. 2023): 10-week-old male C57BL/6 and Nlrp3KO mice were intragastrically administered tabersonine (10 mg/kg) dissolved in 0.5% CMC-Na three times daily. This was followed by intratracheal instillation of LPS (5 mg/kg). After 6h, mice were sacrificed. [2]
- Sepsis Model (Xu et al. 2023): 10-week-old male C57BL/6 and Nlrp3KO mice were injected intraperitoneally with tabersonine (10 mg/kg) or vehicle. This was followed by intraperitoneal injection of E. coli (1×10^9 CFU/mouse) in PBS. Mouse survival was recorded every 6h for 48h. [2]
- HCC Xenograft Model (Li et al. 2024): 2×10^7 HepG2 cells were injected subcutaneously into male BALB/c nude mice. After three days, tabersonine (25 or 50 mg/kg) was administered by gavage daily for three consecutive weeks. Tumor volume was measured every three days. After three weeks, mice were euthanized, and tumors were collected for analysis. [3]

mouse LD50 intravenous 100 mg/kg Annales Pharmaceutiques Francaises., 13(123), 1955 [PMID:14377161]

[1]. Tabersonine attenuates lipopolysaccharide-induced acute lung injury via suppressing TRAF6 ubiquitination. Biochem Pharmacol. 2018 Aug;154:183-192.

- Background: Tabersonine is a natural indole alkaloid from Catharanthus roseus. It is a key precursor in the synthesis of the anticancer drugs vincristine and vinblastine. [1][2][3]
- Mechanism in ALI (Zhang et al. 2018): Tabersonine ameliorates LPS-induced acute lung injury by suppressing the K63-linked polyubiquitination of TRAF6, which in turn inhibits the downstream NF-κB and p38/MK2 signaling pathways, leading to reduced production of pro-inflammatory mediators. [1]
- Mechanism in NLRP3-driven diseases (Xu et al. 2023): Tabersonine is a direct NLRP3 inhibitor. It binds to the NACHT domain of NLRP3, inhibiting its ATPase activity and self-oligomerization, thereby blocking inflammasome assembly, ASC speck formation, and subsequent caspase-1 activation and IL-1β maturation. [2]
- Mechanism in Hepatocellular Carcinoma (Li et al. 2024): Tabersonine induces apoptosis in HepG2 cells through both the mitochondrial pathway (by reducing membrane potential, increasing Bax/Bcl-2 ratio, and releasing cytochrome c) and the death receptor pathway (by upregulating Fas/FasL and activating Caspase-8). It also inhibits the PI3K/Akt signaling pathway. This study is the first to demonstrate these dual apoptotic mechanisms for tabersonine in liver cancer. [3]

C21H24N2O2

372.16

29479-00-3

Tabersonine;4429-63-4

12443187

White to off-white solid at room temperature

4.099

2

4

3

26

669

3

C123C([H])([H])C([H])([H])N4C(C(=C(C(C(C([H])([H])[H])([H])[H])(C([H])([H])C(C(OC([H])([H])[H])=O)=C1N(c1c2c([H])c([H])c([H])c1[H])[H])C34[H])[H])[H])([H])[H]

BBASQSWPQOKOQI-OCIDDWSYSA-N

InChI=1S/C21H24N2O2.ClH/c1-3-20-9-6-11-23-12-10-21(19(20)23)15-7-4-5-8-16(15)22-17(21)14(13-20)18(24)25-2;/h4-9,19,22H,3,10-13H2,1-2H3;1H/t19-,20-,21-;/m0./s1

methyl (1R,12R,19S)-12-ethyl-8,16-diazapentacyclo[10.6.1.01,9.02,7.016,19]nonadeca-2,4,6,9,13-pentaene-10-carboxylate;hydrochloride

Tabersonine Hydrochloride; 29479-00-3; DGR7D6J5TR; Tabersonine monohydrochloride; Aspidospermidine-3-carboxylic acid, 2,3,6,7-tetradehydro-, methyl ester, monohydrochloride, (5alpha,12R,19alpha)-;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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 (134.1 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)