| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
Arvenin I acts as a covalent activator of MKK3 (mitogen-activated protein kinase kinase 3). Chemoproteomic and mechanistic analyses revealed that arvenin I contains an electrophilic reactive functional group that allows it to covalently react with and hyperactivate MKK3. This covalent binding leads to the activation of the downstream p38MAPK signaling pathway, which subsequently revives the mitochondrial fitness of exhausted T cells within the cancer microenvironment. Notably, this mechanism is distinct from other cucurbitacins (such as cucurbitacin B, I, and E), which primarily act as direct cytotoxic agents or JAK2/STAT3 inhibitors. The glycosylation at the C-2 position is critical for this specific MKK3-targeting activity.
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|---|---|
| ln Vitro |
In A-549, HT-29, OVCAR, and MCF-7 cancer cell lines, arvenin I shows moderate activity with IC50 values of 17.0 µM, 49.4 µM, 14.7 µM, and 42.8 µM, respectively [1].
Arvenin I exhibits moderate cytotoxic activity against several human cancer cell lines and demonstrates T cell-activating properties in vitro. In cytotoxicity assays, arvenin I showed IC50 values of 17.0 µM (A-549 lung carcinoma), 49.4 µM (HT-29 colon adenocarcinoma), 14.7 µM (OVCAR ovarian cancer), and 42.8 µM (MCF-7 breast cancer). This moderate cytotoxicity contrasts sharply with other cucurbitacins that exhibit nanomolar potency, highlighting arvenin I's distinct, non-cytotoxic primary mechanism as an immunomodulator. In T cell assays, arvenin I was identified from screening 232 electrophilic natural products as a compound that activates T cells within a cancer-competitive environment. Mechanistic studies confirmed that arvenin I increases mitochondrial oxygen consumption and energy production in exhausted T cells through MKK3-p38MAPK pathway activation. |
| ln Vivo |
Arvenin I demonstrates significant in vivo efficacy in mouse tumor models, enhancing the effectiveness of cancer immunotherapy both as a monotherapy and in combination with immune checkpoint inhibitors. In syngeneic mouse tumor models, administration of arvenin I significantly suppressed tumor growth. When combined with anti-PD-L1 immune checkpoint inhibitors, the antitumor effect was even more pronounced. Mechanistic analysis in vivo confirmed that arvenin I reduces T cell exhaustion and enhances antitumor immune effects by restoring mitochondrial fitness in exhausted CD8+ T cells. These findings position arvenin I as a promising immunomodulatory agent for combination immunotherapy strategies.
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| Enzyme Assay |
A specific chemoproteomic assay protocol was used to identify arvenin I's target as MKK3. The study employed a cell-based screening system that emulates cancer-attenuated T cells to evaluate 232 electrophilic natural products. For target identification, the following approach is typically used: (1) Immobilization of arvenin I on affinity resin (e.g., through a photoaffinity or click chemistry probe); (2) Incubation with cell lysates to allow covalent binding to target proteins; (3) Washing to remove non-specifically bound proteins; (4) Elution and identification of bound proteins by mass spectrometry (LC-MS/MS); (5) Validation of MKK3 as the specific target through competition assays, siRNA knockdown, and recombinant protein binding studies.
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| Cell Assay |
The anti-proliferative activity of arvenin I was evaluated using standard MTT or SRB colorimetric assays on human cancer cell lines. For T cell activation studies, a specialized cell-based system that emulates cancer-attenuated T cells was developed. The typical protocol: (1) Culture A-549, HT-29, OVCAR, or MCF-7 cells in appropriate media (e.g., DMEM or RPMI-1640 with 10% FBS); (2) Seed cells in 96-well plates at densities of 5,000-10,000 cells/well; (3) Allow overnight attachment; (4) Treat with arvenin I at various concentrations (typically 0.1-100 µM) for 48-72 hours; (5) Add MTT reagent (5 mg/mL) and incubate for 4 hours; (6) Dissolve formazan crystals in DMSO and measure absorbance at 570 nm; (7) Calculate IC50 values by regression analysis. For T cell assays, cells are co-cultured with cancer cells or treated with conditioned media to induce exhaustion before arvenin I addition.
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| Animal Protocol |
In vivo studies have been conducted using syngeneic mouse tumor models. The typical protocol: (1) Implant tumor cells (e.g., colon carcinoma or melanoma cells) subcutaneously into immunocompetent mice; (2) Allow tumors to reach a palpable size (approximately 50-100 mm³); (3) Randomize mice into treatment groups (vehicle control, arvenin I alone, anti-PD-L1 alone, and combination); (4) Administer arvenin I via intraperitoneal injection or oral gavage at appropriate doses (formulation: DMSO:PEG300:Tween 80:Saline = 10:40:5:45 or suspension in 0.5% CMC-Na); (5) Continue treatment for 2-3 weeks; (6) Measure tumor volume every 2-3 days using calipers; (7) At study endpoint, collect tumors, draining lymph nodes, and blood for flow cytometry, immunohistochemistry, and cytokine analysis to assess T cell exhaustion markers (e.g., PD-1, TIM-3) and activation status.
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| ADME/Pharmacokinetics |
Based on its physicochemical properties, the compound has a molecular weight of 720.84, a calculated logP of 0.38 (low lipophilicity), a topological polar surface area (TPSA) of 213.4 Ų, with 6 hydrogen bond donors and 13 hydrogen bond acceptors. The molecular complexity is high (1520), which may present challenges for oral absorption. The presence of the glucose moiety significantly increases water solubility compared to cucurbitacin B aglycone. For in vivo formulation, recommended solvents include DMSO for stock solutions, and injection formulations such as DMSO:Tween 80:Saline (10:5:85) or DMSO:PEG300:Tween 80:Saline (10:40:5:45). For oral administration, suspension in 0.5% CMC-Na is recommended.
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| Toxicity/Toxicokinetics |
In the in vivo mouse studies reported in 2025, safety assessments showed that arvenin I did not cause severe adverse effects on mouse body weight or major liver function indicators. This suggests a favorable preliminary safety profile at the doses used for immunotherapy studies. However, comprehensive toxicological data including LD50 values, organ-specific toxicity, genotoxicity, and long-term safety profiles are not yet available. As a cucurbitacin glycoside, arvenin I is expected to have lower direct cytotoxicity compared to cucurbitacin B aglycone due to the glucose modification. It should be emphasized that this compound is strictly for research use only and is not approved for human therapeutic use. Given its covalent binding mechanism, careful handling with appropriate personal protective equipment is recommended.
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| References | |
| Additional Infomation |
Arvenin I is an organic molecular entity. It has been reported to exist in Streptomyces, loofah, and other organisms with relevant data.
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| Molecular Formula |
C38H56O13
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|---|---|
| Molecular Weight |
720.84
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| Exact Mass |
720.372
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| CAS # |
65247-27-0
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| PubChem CID |
6441104
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| Appearance |
Off-white to light yellow solid at room temperature
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
856.0±65.0 °C at 760 mmHg
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| Flash Point |
257.9±27.8 °C
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| Vapour Pressure |
0.0±0.6 mmHg at 25°C
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| Index of Refraction |
1.594
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| LogP |
0.38
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| Hydrogen Bond Donor Count |
6
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| Hydrogen Bond Acceptor Count |
13
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| Rotatable Bond Count |
9
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| Heavy Atom Count |
51
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| Complexity |
1520
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| Defined Atom Stereocenter Count |
14
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| SMILES |
CC(=O)OC(C)(C)/C=C/C(=O)[C@@](C)([C@H]1[C@@H](C[C@@]2(C)[C@@H]3CC=C4[C@@H](C[C@@H](C(=O)C4(C)C)O[C@H]5[C@@H]([C@H]([C@@H]([C@@H](CO)O5)O)O)O)[C@]3(C)C(=O)C[C@]12C)O)O
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| InChi Key |
PQOVWWZVVIGRPP-BBANTJNRSA-N
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| InChi Code |
InChI=1S/C38H56O13/c1-18(40)51-33(2,3)13-12-25(42)38(9,48)30-21(41)15-35(6)24-11-10-19-20(37(24,8)26(43)16-36(30,35)7)14-22(31(47)34(19,4)5)49-32-29(46)28(45)27(44)23(17-39)50-32/h10,12-13,20-24,27-30,32,39,41,44-46,48H,11,14-17H2,1-9H3/b13-12+/t20-,21-,22+,23-,24+,27-,28+,29-,30+,32-,35+,36-,37+,38+/m1/s1
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| Chemical Name |
[(E,6R)-6-hydroxy-6-[(2S,8S,9R,10R,13R,14S,16R,17R)-16-hydroxy-4,4,9,13,14-pentamethyl-3,11-dioxo-2-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-2,7,8,10,12,15,16,17-octahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-5-oxohept-3-en-2-yl] acetate
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| Synonyms |
Arvenin I; 65247-27-0; [(E,6R)-6-hydroxy-6-[(2S,8S,9R,10R,13R,14S,16R,17R)-16-hydroxy-4,4,9,13,14-pentamethyl-3,11-dioxo-2-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-2,7,8,10,12,15,16,17-octahydro-1H-cyclopenta[a]phenanthren-17-yl]-2-methyl-5-oxohept-3-en-2-yl] acetate; ((E,6R)-6-hydroxy-6-((2S,8S,9R,10R,13R,14S,16R,17R)-16-hydroxy-4,4,9,13,14-pentamethyl-3,11-dioxo-2-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl)oxy-2,7,8,10,12,15,16,17-octahydro-1H-cyclopenta(a)phenanthren-17-yl)-2-methyl-5-oxohept-3-en-2-yl) acetate; RefChem:114392;
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| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
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.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] Oral Formulations
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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.3873 mL | 6.9364 mL | 13.8727 mL | |
| 5 mM | 0.2775 mL | 1.3873 mL | 2.7745 mL | |
| 10 mM | 0.1387 mL | 0.6936 mL | 1.3873 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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