-Norcantharidin Chemical Structure.png)
Physicochemical Properties
| Molecular Formula | C8H8O4 |
| Molecular Weight | 168.15 |
| Exact Mass | 168.042 |
| Elemental Analysis | C, 57.14; H, 4.80; O, 38.06 |
| CAS # | 29745-04-8 |
| Related CAS # | 129-67-9 (disodium); 17439-94-0 (diammonium); 29745-04-8 |
| PubChem CID | 93004 |
| Appearance | Solid powder |
| Density | 1.5±0.1 g/cm3 |
| Boiling Point | 362.5±35.0 °C at 760 mmHg |
| Melting Point | 114-116ºC |
| Flash Point | 167.0±26.0 °C |
| Vapour Pressure | 0.0±0.8 mmHg at 25°C |
| Index of Refraction | 1.550 |
| LogP | -0.85 |
| Hydrogen Bond Donor Count | 0 |
| Hydrogen Bond Acceptor Count | 4 |
| Rotatable Bond Count | 0 |
| Heavy Atom Count | 12 |
| Complexity | 246 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | O1C2([H])C([H])([H])C([H])([H])C1([H])C1([H])C(=O)OC(C12[H])=O |
| InChi Key | JAABVEXCGCXWRR-FBXFSONDSA-N |
| InChi Code | InChI=1S/C8H8O4/c9-7-5-3-1-2-4(11-3)6(5)8(10)12-7/h3-6H,1-2H2/t3-,4+,5-,6+ |
| Chemical Name | (1R,2S,6R,7S)-4,10-dioxatricyclo[5.2.1.02,6]decane-3,5-dione |
| Synonyms | Endothall anhydride;Norcantharidin |
| 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) |
Biological Activity
| Targets | EGFR; c-Met |
| ln Vitro |
NCTD induced cell cycle arrest at G2/M phase in both cell lines. The early and late apoptosis was also observed. Further investigation indicated that NCTD suppressed not only the expression of the total EGFR and the phosphorylated EGFR but also the expression of the total c-Met and the phosphorylated c-Met in colon cancer cells. Moreover, EGFR expression could be mostly restored by co-treatment with MG132, a proteasome inhibitor. In addition, NCTD-induced cell death was comparable to that of the anti-cancer drug gefitinib, a tyrosine kinase inhibitor for EGFR, based on the immunoblot analysis of the expressed proteins after the drug treatment [1]. NCTD dose and time dependently inhibits HMC proliferation significantly (p < .05). Apoptosis dose and time dependently increased after NCTD treatment. Cell-cycle analysis revealed that the number of cells in the G2 phase increased significantly, whereas the fraction of cells in the S phase decreased, especially 24 h after 5 μg/ml NCTD treatment [3]. |
| ln Vivo | To help treat cancer patients, NCTD may be a practical and affordable medication candidate to replace gefitinib[1].In nude mice, NCTD prevents the growth of human gallbladder carcinoma xenografted tumors by causing apoptosis and interrupting the cell cycle in vivo[2]. |
| Enzyme Assay |
Western blot analysis [1] HCT116 and HT29 cells were cultured in 10-cm dishes for 24 h before treated with different concentrations of NCTD or NCTD plus MG132. After being incubated with another 72 h, the cells were collected with cell-scrapers into 1.5 mL tube and put on ice for 30 min in whole cell lysis buffer containing protease inhibitors. Proteins were quantified by BCA protein assay kit. Equal amount of proteins from different sample (50 μg) were resolved over 8 or 12% SDS-polyacrylamide gel by electrophoresis and then transferred to nitrocellulose membrane. The membranes were put in blocking buffer at room temperature for 2 h before appropriate primary antibody was added. The membrane was incubated in the presence of primary antibody at 4 °C overnight before the corresponding secondary antibodies were added. The membranes were then visualized by using Western Lightning. |
| Cell Assay |
HCT116 and HT29, two human colon cancer cell lines, were employed as model systems to study the anti-cancer molecular mechanism of NCTD. Using flow cytometry, cell cycle arrest and early/late apoptosis were examined. Western blot analysis was used to measure the amounts of EGFR, phospho-EGFR, c-Met, phospho-c-Met, and other related proteins[1]. HMC cells were divided into a normal control group, and various concentrations of NCTD group (2.5, 5, 10, 20, or 40 μg/mL). Cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, apoptosis was detected by Annexin V/propidium iodide (PI) assays, and morphological analysis was performed by Hoechest 33258 staining. Finally, cell cycle was analyzed by flow cytometry [3]. |
| Animal Protocol | Subcutaneous GBC-SD cells were used to create the tumor xenograft model of human gallbladder carcinoma in nude mice in vivo. The 5-FU, NCTD, control, and NCTD+5-FU groups of experimental mice were randomly assigned to receive various treatments. The size, growth curve, and inhibitory rate of the tumor were assessed. Light/electron microscopy and flow cytometry were used to evaluate the xenografted tumors' morphological changes, apoptosis, and cell cycle. RT-PCR and the streptavidin-biotin complex (SABC) method were used to determine the expression of the proteins related to apoptosis, Bcl-2, Bax, and survivin, as well as the cell cycle-related proteins cyclin-D1 and p27[2]. |
| References |
[1]. Norcantharidin Inhibits cell growth by suppressing the expression and phosphorylation of both EGFR and c-Met in human colon cancer cells[J]. BMC Cancer, 2017, 17(1):55. [2]. Norcantharidin Inhibits Growth of Human Gallbladder Carcinoma Xenografted Tumors in Nude Mice by Inducing Apoptosis and Blocking the Cell Cycle in vivo[J]. Hepatobiliary & Pancreatic Diseases International, 2010, 9(4):414-422. [3]. Effect of norcantharidin on the proliferation, apoptosis, and cell cycle of human mesangial cells. Ren Fail. 2017 Nov;39(1):458-464 |
| Additional Infomation | NORCANTHARIDIN is a furofuran. |
Solubility Data
| Solubility (In Vitro) | DMSO: ~33 mg/mL (~196.25 mM) |
| 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 (e.g. IP/IV/IM/SC) 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). 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) 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). 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 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 5.9471 mL | 29.7354 mL | 59.4707 mL | |
| 5 mM | 1.1894 mL | 5.9471 mL | 11.8941 mL | |
| 10 mM | 0.5947 mL | 2.9735 mL | 5.9471 mL |