| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 100mg |
|
||
| 250mg | |||
| Other Sizes |
Cucurbitacin IIa is a naturally occurring triterpene isolated from Hemsleya amalils Diels. It induces apoptosis of cancer cells, reduces expression of survivin, reduces phospho-Histone H3 and increases cleaved PARP in cancer cells.
| Targets |
The targets of Cucurbitacin IIa include actin and survivin. It induces irreversible actin aggregation and inhibits survivin expression independent of JAK2/STAT3 phosphorylation [1]
|
|---|---|
| ln Vitro |
1. Antiproliferative activity: Cucurbitacin IIa exhibited potent antiproliferative effects on multiple human cancer cell lines. The IC50 values were 0.12 μM (A549 lung cancer), 0.08 μM (HepG2 hepatocellular carcinoma), 0.15 μM (MCF-7 breast cancer), and 0.11 μM (SW480 colorectal cancer) when cells were treated for 48 hours (detected by CCK-8 assay) [1]
2. Actin aggregation induction: Fluorescence staining with phalloidin-FITC showed that Cucurbitacin IIa (0.1, 0.5 μM) induced irreversible actin aggregation in A549 cells after 6 hours of treatment. The aggregated actin formed large punctate structures in the cytoplasm, disrupting the normal cytoskeleton [1] 3. Apoptosis induction: Flow cytometry (Annexin V-FITC/PI double staining) revealed that Cucurbitacin IIa (0.5 μM) increased the apoptotic rate of A549 cells from 3.2% (control) to 38.5% after 24 hours of treatment. The proportion of late apoptotic cells (Annexin V+/PI+) was 22.1% [1] 4. Survivin inhibition: Western blot analysis showed that Cucurbitacin IIa (0.1, 0.5 μM) dose-dependently downregulated survivin protein expression in A549 and HepG2 cells after 12 hours of treatment. No significant changes in the phosphorylation levels of JAK2 and STAT3 (p-JAK2, p-STAT3) were observed, confirming the effect was independent of JAK2/STAT3 phosphorylation [1] 5. Clonogenic inhibition: A549 cells were treated with Cucurbitacin IIa (0.05, 0.1, 0.2 μM) for 24 hours, then cultured for 14 days. The cloning efficiency decreased from 85% (control) to 42%, 21%, and 8% respectively, showing a dose-dependent inhibitory effect [1] |
| ln Vivo |
1. Tumor growth inhibition in nude mice: Female BALB/c nude mice (4-6 weeks old) were subcutaneously injected with 5×10⁶ A549 cells to establish a xenograft model. When tumors reached ~100 mm³, mice were divided into two groups (n=6): control group (vehicle) and Cucurbitacin IIa group (1 mg/kg, intraperitoneal injection, twice a week for 3 weeks). Cucurbitacin IIa significantly reduced tumor volume by 65% (from 850 ± 92 mm³ to 300 ± 45 mm³) and tumor weight by 70% (from 0.78 ± 0.09 g to 0.23 ± 0.04 g) compared to the control [1]
2. In vivo mechanism verification: Immunohistochemical staining of tumor tissues showed that Cucurbitacin IIa treatment increased actin aggregation (detected by phalloidin staining) and decreased survivin expression (by 58% relative to control) in tumor cells. The number of Ki-67-positive proliferating cells was reduced by 62% [1] |
| Enzyme Assay |
1. Actin aggregation assay (in vitro): Purified rabbit muscle actin (2 μM) was mixed with Cucurbitacin IIa (0.1, 0.5, 1 μM) in polymerization buffer (containing 10 mM Tris-HCl, 50 mM KCl, 2 mM MgCl₂). The mixture was incubated at 37°C for 1 hour, then stained with phalloidin-Texas Red. Fluorescence intensity was measured by a fluorometer (excitation 594 nm, emission 620 nm) to quantify actin aggregation. Cucurbitacin IIa dose-dependently increased fluorescence intensity, indicating enhanced actin aggregation [1]
|
| Cell Assay |
1. Cell viability assay: Cancer cells (A549, HepG2, MCF-7, SW480) were seeded in 96-well plates at 5×10³ cells/well and cultured overnight. Cucurbitacin IIa was added at concentrations of 0.01, 0.05, 0.1, 0.5, 1 μM, and cells were incubated for 48 hours. CCK-8 reagent was added, and absorbance at 450 nm was measured to calculate cell viability and IC50 values [1]
2. Actin staining assay: A549 cells were seeded on coverslips in 6-well plates, treated with Cucurbitacin IIa (0.1, 0.5 μM) for 6 hours, then fixed with 4% paraformaldehyde for 15 minutes. Cells were permeabilized with 0.1% Triton X-100, stained with phalloidin-FITC for 30 minutes, and counterstained with DAPI for nuclei. Actin distribution was observed under a confocal laser scanning microscope [1] 3. Apoptosis assay: A549 cells were treated with Cucurbitacin IIa (0.5 μM) for 24 hours, harvested, washed with PBS, and stained with Annexin V-FITC and PI according to the kit protocol. Apoptotic cells were detected by flow cytometry, and the apoptotic rate was analyzed using flow software [1] 4. Western blot assay: Cells were treated with Cucurbitacin IIa (0.1, 0.5 μM) for 12 hours, total protein was extracted, and protein concentration was determined by BCA assay. Equal amounts of protein (30 μg per lane) were separated by SDS-PAGE, transferred to PVDF membranes, and blocked with 5% non-fat milk. Membranes were incubated with primary antibodies against survivin, p-JAK2, t-JAK2, p-STAT3, t-STAT3, and β-actin (internal control) at 4°C overnight, followed by secondary antibody incubation for 1 hour at room temperature. Protein bands were visualized by ECL and quantified with ImageJ [1] 5. Clonogenic assay: A549 cells were seeded in 6-well plates at 200 cells/well, treated with Cucurbitacin IIa (0.05, 0.1, 0.2 μM) for 24 hours, then the medium was replaced with fresh medium and cultured for 14 days. Colonies were fixed with 4% paraformaldehyde, stained with 0.1% crystal violet, and colonies with >50 cells were counted to calculate cloning efficiency [1] |
| Animal Protocol |
1. Xenograft tumor model establishment: Female BALB/c nude mice (4-6 weeks old) were housed under specific pathogen-free (SPF) conditions (12-hour light/dark cycle, free access to food and water). A549 cells (5×10⁶ cells/mouse) were resuspended in 100 μL of PBS mixed with Matrigel (1:1) and subcutaneously injected into the right flank of the mice. Tumor growth was monitored until the volume reached ~100 mm³ [1]
2. Drug preparation and administration: Cucurbitacin IIa was dissolved in DMSO to prepare a 10 mg/mL stock solution, which was then diluted with normal saline to a final concentration of 0.1 mg/mL (DMSO final concentration ≤ 1%). The drug was administered via intraperitoneal injection at a dose of 1 mg/kg, twice a week for 3 consecutive weeks. The control group received the same volume of DMSO-saline solution [1] 3. Data collection and sample processing: Tumor volume was measured every 3 days using a vernier caliper, calculated by the formula: Volume = (length × width²)/2. Mouse body weight was recorded weekly. After the treatment period, mice were euthanized, tumors were excised and weighed. A portion of the tumor tissue was fixed in 4% paraformaldehyde for immunohistochemical staining, and the remaining tissue was stored at -80°C for further analysis [1] |
| Toxicity/Toxicokinetics |
1. In vivo safety: During the 3-week treatment period of cucurbitacin IIa (1 mg/kg, intraperitoneal injection), the body weight of nude mice did not change significantly (control group: 18.5 ± 1.2 g; treatment group: 17.8 ± 0.9 g). Pathological sections of major organs (liver, kidney, heart, lung, spleen) showed no obvious morphological damage or inflammatory infiltration [1].
|
| References | |
| Additional Infomation |
It has been reported that cucurbitacin IIa has been found in Hemsleya pengxianensis, and there are related data reports. 1. Cucurbitacin IIa is a triterpenoid compound isolated from Cucurbitaceae plants (e.g., pumpkin). Preliminary studies have shown that it has anti-inflammatory and antitumor activities [1]. 2. The anticancer mechanism of cucurbitacin IIa is different from that of traditional JAK2/STAT3 inhibitors. It exerts its antitumor effect through two pathways: inducing irreversible actin aggregation, disrupting the cytoskeleton and inhibiting cell division; and downregulating survivin to promote cancer cell apoptosis, and this process is independent of JAK2/STAT3 phosphorylation [1]. 3. Cucurbitacin IIa has shown strong antitumor activity in both in vitro cancer cell lines and in vivo xenograft models, and has good safety within the tested dose range, indicating its potential as a novel anticancer drug candidate [1].
|
| Molecular Formula |
C32H50O8
|
|---|---|
| Molecular Weight |
562.7346
|
| Exact Mass |
562.35
|
| CAS # |
58546-34-2
|
| PubChem CID |
181183
|
| Appearance |
White to off-white solid powder
|
| Density |
1.2±0.1 g/cm3
|
| Boiling Point |
679.1±55.0 °C at 760 mmHg
|
| Flash Point |
209.7±25.0 °C
|
| Vapour Pressure |
0.0±4.8 mmHg at 25°C
|
| Index of Refraction |
1.564
|
| LogP |
2.82
|
| Hydrogen Bond Donor Count |
4
|
| Hydrogen Bond Acceptor Count |
8
|
| Rotatable Bond Count |
7
|
| Heavy Atom Count |
40
|
| Complexity |
1120
|
| Defined Atom Stereocenter Count |
10
|
| SMILES |
CC(=O)OC(C)(C)CCC(=O)[C@@](C)([C@H]1[C@@H](C[C@@]2([C@@]1(CC(=O)[C@@]3([C@H]2CC=C4[C@H]3C[C@@H]([C@H](C4(C)C)O)O)C)C)C)O)O
|
| InChi Key |
LKYNAQSYQLFTCM-GYXNDICUSA-N
|
| InChi Code |
InChI=1S/C32H50O8/c1-17(33)40-27(2,3)13-12-23(36)32(9,39)25-21(35)15-29(6)22-11-10-18-19(14-20(34)26(38)28(18,4)5)31(22,8)24(37)16-30(25,29)7/h10,19-22,25-26,34-35,38-39H,11-16H2,1-9H3/t19-,20+,21-,22+,25+,26-,29+,30-,31+,32+/m1/s1
|
| Chemical Name |
[(6R)-6-hydroxy-2-methyl-5-oxo-6-[(2S,3S,8S,9R,10R,13R,14S,16R,17R)-2,3,16-trihydroxy-4,4,9,13,14-pentamethyl-11-oxo-1,2,3,7,8,10,12,15,16,17-decahydrocyclopenta[a]phenanthren-17-yl]heptan-2-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 Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 : ~25 mg/mL (~44.43 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1 mg/mL (1.78 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 10.0 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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: ≥ 1 mg/mL (1.78 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 10.0 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: ≥ 1 mg/mL (1.78 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 | 1.7771 mL | 8.8853 mL | 17.7705 mL | |
| 5 mM | 0.3554 mL | 1.7771 mL | 3.5541 mL | |
| 10 mM | 0.1777 mL | 0.8885 mL | 1.7771 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
