Flavokawain B modulates multiple signaling pathways, including NF-κB, PI3K/Akt, and MAPK, which are critical for cell survival, proliferation, and inflammation. By inhibiting NF-κB signaling, the compound reduces the expression of anti-apoptotic genes and promotes apoptosis. It also inhibits the PI3K/Akt pathway, which is involved in cell survival and growth. The compound's effects on the MAPK pathway, including JNK and p38, contribute to its pro-apoptotic and anti-inflammatory activities. Flavokawain B activates caspase-9, -3, and -8, leading to the cleavage of PARP and the induction of apoptosis. It downregulates the anti-apoptotic protein Bcl-2 while upregulating the pro-apoptotic protein Bax, shifting the balance towards apoptosis. The compound also promotes the generation of reactive oxygen species (ROS), which can contribute to its cytotoxic effects. Additionally, Flavokawain B inhibits MMP-9, an enzyme involved in extracellular matrix degradation and cancer metastasis. Its ability to target protein neddylation, a post-translational modification that regulates the activity of cullin-RING ubiquitin ligases, enhances the anti-prostate cancer effect of Bortezomib via Skp2 degradation. The compound's multi-targeted activity makes it a valuable tool for studying the complex signaling networks involved in cancer and inflammation.

Flavokawain B exhibits potent in vitro activity against various cancer cell lines, including prostate, breast, colon, and lung cancer cells. It induces apoptosis through the activation of caspases and the modulation of Bcl-2 family proteins. The compound inhibits cell proliferation and colony formation in a dose-dependent manner. It also inhibits the migration and invasion of cancer cells, potentially through the inhibition of MMP-9. Flavokawain B has strong antiangiogenic activity, as it inhibits the migration and tube formation of human brain endothelial cells (HUVEC) at very low non-toxic concentrations. This antiangiogenic effect is important for its anticancer activity, as it limits the blood supply to tumors. The compound also inhibits the production of inflammatory mediators, such as nitric oxide (NO) and prostaglandin E2 (PGE2), in LPS-induced RAW 264.7 macrophages, demonstrating its anti-inflammatory activity. Flavokawain B's in vitro activity is well-characterized and forms the basis for its use in cancer and inflammation research. The compound's potency against various cancer cell lines and its ability to modulate multiple signaling pathways make it a promising candidate for further development as a therapeutic agent.

Flavokawain B demonstrates oral efficacy in murine cancer models, including xenograft models of prostate cancer. In these models, oral administration of the compound leads to significant tumor growth inhibition. The compound's oral bioavailability is a key advantage for its potential therapeutic use. In addition to its antitumor activity, Flavokawain B has shown anti-inflammatory activity in vivo. It reduces inflammation in animal models of inflammatory diseases, likely through its inhibition of NF-κB and other inflammatory pathways. The compound's antiangiogenic activity has also been demonstrated in vivo, where it inhibits the formation of new blood vessels in Matrigel plug or corneal angiogenesis assays. The in vivo efficacy of Flavokawain B is attributed to its ability to modulate multiple signaling pathways, induce apoptosis, and inhibit angiogenesis and inflammation. The compound's safety and efficacy profiles have been evaluated in preclinical studies, and it has shown promising results. However, Flavokawain B is a known hepatotoxic constituent from kava root, and it induces GSH-sensitive oxidative stress. This hepatotoxicity is a concern for its potential therapeutic use and highlights the need for careful safety evaluation. The compound's diverse pharmacological effects, coupled with its oral bioavailability, make it a promising candidate for further development as a therapeutic agent, particularly in the areas of cancer and inflammation.

Cell-free assays with Flavokawain B are used to assess its effects on various molecular targets, including kinases, caspases, and transcription factors. For kinase inhibition, purified kinases (e.g., PI3K, Akt, MAPK) are incubated with ATP and peptide substrates in the presence of varying concentrations of Flavokawain B. Kinase activity is measured by radioactive phosphate incorporation or luminescent ADP detection to determine IC50 values. For caspase activation, purified caspases (e.g., caspase-3, -8, -9) are incubated with fluorogenic substrates in the presence of the compound, and the cleavage of the substrate is measured fluorometrically. For NF-κB inhibition, cell-free assays using purified NF-κB proteins or DNA-binding assays are performed to assess the compound's ability to inhibit NF-κB DNA binding activity. These cell-free assays are essential for characterizing the direct effects of Flavokawain B on its molecular targets and for understanding its mechanism of action. The compound's ability to modulate multiple targets is confirmed through these assays, providing a mechanistic basis for its diverse pharmacological activities. The results from cell-free assays are complemented by cellular and in vivo studies to provide a comprehensive picture of the compound's biological activity.

In cellular assays, Flavokawain B is used to study its effects on various cell types, including cancer cells, endothelial cells, and immune cells. Cancer cell lines (e.g., prostate, breast, colon, lung) are treated with the compound at various concentrations, typically ranging from 1 to 100 µM. Cell viability is assessed using MTT or CellTiter-Glo assays. Apoptosis is evaluated by flow cytometry using Annexin V/PI staining, by measuring caspase activity, or by Western blot analysis of apoptosis-related proteins (e.g., cleaved PARP, Bcl-2, Bax). Cell cycle analysis is performed by flow cytometry. The compound's effects on cell migration and invasion are assessed using Transwell or wound healing assays. Its antiangiogenic activity is evaluated in HUVEC assays, where the compound's effects on cell migration and tube formation are assessed. The compound's anti-inflammatory activity is assessed in macrophage cell lines (e.g., RAW 264.7), where its effects on the production of inflammatory mediators (e.g., NO, PGE2, cytokines) are measured by ELISA or Griess assay. These cellular assays are crucial for understanding the functional consequences of Flavokawain B treatment and for validating its activity as an anticancer and anti-inflammatory agent.

In vivo efficacy of Flavokawain B is evaluated in mouse xenograft models of human cancers, as well as in models of inflammatory diseases. In xenograft models, cancer cells are implanted subcutaneously into immunodeficient mice. When tumors reach a certain size, Flavokawain B is administered orally or intraperitoneally. Tumor growth is monitored by caliper measurements. At the end of the study, tumors are harvested for histological analysis, immunohistochemistry, and molecular analysis. The compound's effects on apoptosis, angiogenesis, and proliferation are assessed in tumor tissues. In inflammatory models, the compound is administered to animals with induced inflammation (e.g., carrageenan-induced paw edema), and the reduction in inflammation is measured. The compound's antiangiogenic activity is evaluated in Matrigel plug or corneal angiogenesis assays, where its ability to inhibit the formation of new blood vessels is assessed. These in vivo studies are essential for confirming the compound's efficacy in a physiologically relevant context and for guiding the development of Flavokawain B as a therapeutic agent. The compound's oral bioavailability and its ability to modulate multiple signaling pathways make it a promising candidate for further development. However, its hepatotoxicity is a concern that must be addressed through careful safety evaluation and formulation strategies.

Flavokawain B is orally bioavailable, a key advantage for its potential therapeutic use. Pharmacokinetic studies in rodents determine parameters including bioavailability, half-life, and tissue distribution following oral or intravenous administration. The compound is absorbed from the gastrointestinal tract and distributed to various tissues, including tumors. Its oral bioavailability allows for convenient administration in preclinical studies. The compound's metabolism and clearance pathways are characterized to understand its elimination from the body. The pharmacokinetic data obtained from these studies are essential for designing appropriate dosing regimens and for interpreting the results of efficacy and toxicity studies. The compound's ability to reach therapeutic concentrations in target tissues is a critical factor for its efficacy. The pharmacokinetic profile of Flavokawain B is influenced by factors such as the formulation, the dose, and the animal species. Further studies are needed to fully characterize the compound's pharmacokinetics and to optimize its delivery for therapeutic applications.

Flavokawain B is a known hepatotoxic constituent from kava root, and it induces GSH-sensitive oxidative stress. Hepatotoxicity has been reported in association with the use of kava products, and Flavokawain B is considered one of the hepatotoxic constituents. The compound's toxicity is attributed to its ability to deplete cellular glutathione (GSH) and induce oxidative stress, leading to liver cell damage. In preclinical studies, the compound's hepatotoxicity is evaluated by measuring liver enzyme levels (e.g., ALT, AST) and by histological examination of liver tissue. The compound's toxicity may be dose-dependent, and high doses may cause significant liver damage. In addition to hepatotoxicity, Flavokawain B may have other toxic effects, including gastrointestinal disturbances and hematological changes. The compound's safety profile must be carefully evaluated in preclinical studies to determine the therapeutic window and to identify potential risks. The compound is intended for research use only and is not approved for human therapeutic use. Standard safety precautions should be followed when handling Flavokawain B, including the use of appropriate personal protective equipment and adherence to institutional safety guidelines.

[1]. The In Vitro and In Vivo Antiangiogenic Effects of Flavokawain B. Phytother Res. 2017 Oct;31(10):1607-1613.

[2]. Flavokawain B targets protein neddylation for enhancing the anti-prostate cancer effect of Bortezomib via Skp2 degradation. Cell Commun Signal. 2019 Mar 18;17(1):25.

Flavonoid carvain B belongs to the chalcone class of compounds. Its structure is trans-chalcone, with a hydroxyl group at the 2' position and methoxy groups at the 4' and 6' positions. It has been isolated from plants of the genus Piper (such as Piper methysticum and Piper rusbyi) and possesses anti-leishmaniatic, anti-inflammatory, and antitumor activities. It functions as a metabolite, an anti-leishmaniatic agent, an anti-inflammatory agent, an apoptosis inducer, and an antitumor agent. It belongs to the chalcone, dimethoxybenzene, and phenolic compounds. Its function is related to trans-chalcone.
Flavone carvain B has been reported to exist in Cedrelopsis grevei, Alpinia mutica, and several other organisms with relevant data.
See also: Roots (parts) of Piper methysticum.
Flavokawain B (Flavokavain B, CAS 1775-97-9) is a research-grade compound exclusively for laboratory use. It is an orally active chalcone naturally isolated from the kava plant (Piper methysticum). The compound exhibits potent anti-inflammatory, anticancer, antiangiogenic, and antifungal activities. It is a potent apoptosis inducer that inhibits the growth of various cancer cell lines. Flavokawain B activates caspase-9, -3, and -8, and cleaves PARP, while downregulating Bcl-2 and increasing Bax levels. It inhibits NF-κB, PI3K/Akt, and MAPK signaling pathways, and promotes ROS generation. The compound also inhibits MMP-9. Flavokawain B has strong antiangiogenic activity and inhibits the migration and vascular formation of HUVEC cells at very low non-toxic concentrations. It induces apoptosis in androgen receptor-negative, hormone-refractory prostate cancer cell lines and reduces tumor growth in preclinical models. The compound also targets protein neddylation for enhancing the anti-prostate cancer effect of Bortezomib via Skp2 degradation. Flavokawain B is a known hepatotoxic constituent from kava root, and it induces GSH-sensitive oxidative stress. The compound is not approved for human therapeutic use and is intended for research purposes only. It is available from various commercial suppliers as a high-purity reagent for biochemical and cell-based assays. The compound should be stored according to the manufacturer's recommendations, typically at -20°C, to ensure stability. When handling Flavokawain B, researchers should follow standard safety protocols for handling chemical reagents, including the use of appropriate personal protective equipment and working in a well-ventilated area. The compound is soluble in DMSO and other organic solvents, and stock solutions should be prepared fresh or stored in aliquots to avoid repeated freeze-thaw cycles.

C17H16O4

284.31

284.104

1775-97-9

5356121

Light yellow to yellow solid powder

1.203±0.06 g/cm3

500.1±50.0 °C at 760 mmHg

178-179 ºC (ethanol )

185.8±23.6 °C

0.0±1.3 mmHg at 25°C

1.614

4.01

1

4

5

21

360

0

COC1=CC(=C(C(=C1)OC)C(=O)/C=C/C2=CC=CC=C2)O

QKQLSQLKXBHUSO-CMDGGOBGSA-N

InChI=1S/C17H16O4/c1-20-13-10-15(19)17(16(11-13)21-2)14(18)9-8-12-6-4-3-5-7-12/h3-11,19H,1-2H3/b9-8+

(E)-1-(2-hydroxy-4,6-dimethoxyphenyl)-3-phenylprop-2-en-1-one

FlavokawainB; Flavokawain-B; Flavokawain B

2934.99.9001

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.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~5 mg/mL (~17.59 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.)