-Flavokawain A Chemical Structure.png)
Physicochemical Properties
| Molecular Formula | C18H18O5 |
| Molecular Weight | 314.3325 |
| Exact Mass | 314.115 |
| CAS # | 37951-13-6 |
| Related CAS # | Flavokawain A;3420-72-2 |
| PubChem CID | 5355469 |
| Appearance | Light yellow to yellow solid powder |
| Density | 1.2±0.1 g/cm3 |
| Boiling Point | 529.9±50.0 °C at 760 mmHg |
| Melting Point | 114℃ |
| Flash Point | 193.2±23.6 °C |
| Vapour Pressure | 0.0±1.4 mmHg at 25°C |
| Index of Refraction | 1.600 |
| LogP | 3.96 |
| Hydrogen Bond Donor Count | 1 |
| Hydrogen Bond Acceptor Count | 5 |
| Rotatable Bond Count | 6 |
| Heavy Atom Count | 23 |
| Complexity | 400 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | COC1=CC=C(C=C1)/C=C/C(=O)C2=C(C=C(C=C2OC)OC)O |
| InChi Key | CGIBCVBDFUTMPT-RMKNXTFCSA-N |
| InChi Code | InChI=1S/C18H18O5/c1-21-13-7-4-12(5-8-13)6-9-15(19)18-16(20)10-14(22-2)11-17(18)23-3/h4-11,20H,1-3H3/b9-6+ |
| Chemical Name | (E)-1-(2-hydroxy-4,6-dimethoxyphenyl)-3-(4-methoxyphenyl)prop-2-en-1-one |
| 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) |
Biological Activity
| ln Vitro |
- In human bladder cancer cell lines (T24, 5637, J82), (E)-Flavokawain A exhibited dose-dependent antiproliferative activity. The IC50 values were 8.2 μM (T24), 7.5 μM (5637), and 9.1 μM (J82) after 48 hours of treatment, as determined by MTT assay [1] - (E)-Flavokawain A (5, 10, 20 μM) induced apoptosis in T24 cells in a dose-dependent manner. At 20 μM, the apoptotic rate was 45.3% (detected by TUNEL assay), significantly higher than the control group (3.2%). It upregulated the expression of pro-apoptotic protein Bax (2.8-fold at 20 μM) and downregulated anti-apoptotic protein Bcl-2 (0.3-fold at 20 μM) [1] - The compound triggered mitochondria-dependent apoptotic pathway: it reduced mitochondrial membrane potential (ΔΨm) by 60% at 20 μM (detected by JC-1 staining), promoted cytochrome c release from mitochondria to cytoplasm (1.9-fold increase in cytoplasmic cytochrome c at 20 μM), and activated caspase-9 (2.5-fold) and caspase-3 (3.2-fold) at 20 μM, as measured by Western blot [1] - (E)-Flavokawain A (10, 20 μM) inhibited the colony formation ability of T24 cells: the number of colonies was reduced by 52% and 78% at 10 μM and 20 μM, respectively, compared with the control group [1] |
| ln Vivo |
- In nude mice bearing T24 bladder cancer xenografts, intraperitoneal injection of (E)-Flavokawain A (20 mg/kg, once every 2 days for 3 weeks) significantly suppressed tumor growth. The final tumor volume in the treatment group was 286 ± 45 mm³, which was 62% smaller than the vehicle control group (752 ± 68 mm³). The tumor weight was also reduced by 58% (0.32 ± 0.05 g vs. 0.76 ± 0.08 g in control) [1] - TUNEL staining of tumor tissues showed that the apoptotic index in the (E)-Flavokawain A-treated group was 32.5 ± 4.2%, significantly higher than the control group (5.8 ± 1.1%). Western blot analysis of tumor tissues revealed upregulated Bax expression (2.1-fold) and downregulated Bcl-2 expression (0.4-fold) [1] |
| Cell Assay |
- MTT antiproliferative assay: Bladder cancer cells (T24, 5637, J82) were seeded in 96-well plates at a density of 5×10³ cells/well and cultured overnight. Cells were treated with (E)-Flavokawain A (0, 2.5, 5, 10, 20, 40 μM) for 48 hours. Then, 20 μL MTT solution (5 mg/mL) was added to each well and incubated for 4 hours at 37°C. The medium was removed, and 150 μL DMSO was added to dissolve the formazan crystals. The absorbance was measured at 570 nm, and IC50 values were calculated [1] - Apoptosis detection by TUNEL assay: T24 cells were seeded on coverslips in 6-well plates, treated with (E)-Flavokawain A (5, 10, 20 μM) for 24 hours, fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100, and incubated with TUNEL reaction mixture for 1 hour at 37°C. DAPI was used to stain cell nuclei. The apoptotic rate was calculated by counting TUNEL-positive cells under a fluorescence microscope [1] - Mitochondrial membrane potential and apoptotic protein detection: T24 cells were treated with (E)-Flavokawain A (20 μM) for 12 hours. For ΔΨm detection, cells were stained with JC-1 dye for 30 minutes at 37°C, and fluorescence intensity was measured by flow cytometry. For protein analysis, cells were lysed, and mitochondrial and cytoplasmic fractions were separated by centrifugation. Western blot was performed to detect Bax, Bcl-2, cytochrome c, caspase-9, and caspase-3 expression (β-actin as internal control) [1] - Colony formation assay: T24 cells were seeded in 6-well plates at 2×10³ cells/well, treated with (E)-Flavokawain A (10, 20 μM) for 24 hours, then the medium was replaced with fresh medium and cultured for 14 days. Colonies were fixed with methanol, stained with crystal violet, and counted [1] |
| Animal Protocol |
- Nude mouse xenograft model: Female BALB/c nude mice (4–6 weeks old) were subcutaneously injected with 5×10⁶ T24 bladder cancer cells into the right flank to establish tumor xenografts. When tumors reached a volume of ~100 mm³, mice were randomly divided into two groups (n=6/group): (E)-Flavokawain A treatment group and vehicle control group. The compound was dissolved in DMSO (5%) and diluted with corn oil (95%) to a concentration of 10 mg/mL. The treatment group received intraperitoneal injection of 20 mg/kg (E)-Flavokawain A once every 2 days for 3 weeks, while the control group received the same volume of DMSO/corn oil mixture. Tumor volume was measured every 3 days using a caliper (volume = length × width² / 2). At the end of the experiment, mice were sacrificed, tumors were excised, weighed, and stored at -80°C for further protein and histological analysis [1] |
| Toxicity/Toxicokinetics |
- In the in vivo study, (E)-Flavokawain A (20 mg/kg, ip) did not cause significant changes in mouse body weight (final body weight: 18.2 ± 1.3 g in treatment group vs. 19.1 ± 1.2 g in control group). Histopathological examination of major organs (liver, kidney, heart, lung, spleen) showed no obvious toxic lesions [1] - No in vitro cytotoxicity was observed in normal human bladder epithelial cells (SV-HUC-1) at concentrations up to 20 μM: cell viability remained above 85% compared with the control group [1] |
| References |
[1]. Flavokawain A, a novel chalcone from kava extract, induces apoptosis in bladder cancer cells by involvement of Bax protein-dependent and mitochondria-dependent apoptotic pathway and suppresses tumor growth in mice. Cancer Res. 2005 Apr 15;65(. |
| Additional Infomation |
Flavokawain A is a member of chalcones. 2'-Hydroxy-4,4',6'-trimethoxychalcone has been reported in Boesenbergia rotunda, Vitex quinata, and other organisms with data available. See also: Piper methysticum root (part of). - (E)-Flavokawain A is a novel chalcone isolated from kava (Piper methysticum) extract [1] - Its anticancer effect is mediated through Bax protein-dependent and mitochondria-dependent apoptotic pathways, making it a potential candidate for the treatment of bladder cancer [1] |
Solubility Data
| Solubility (In Vitro) | DMSO : ~25 mg/mL (~79.53 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 | 3.1814 mL | 15.9068 mL | 31.8137 mL | |
| 5 mM | 0.6363 mL | 3.1814 mL | 6.3627 mL | |
| 10 mM | 0.3181 mL | 1.5907 mL | 3.1814 mL |