1. NLRP3 inflammasome (inhibits activation via AhR/Nrf2/NQO1 pathway, EC50 = 12.5 μM for NLRP3-mediated IL-1β secretion inhibition in bone marrow-derived macrophages (BMDMs)) [2]
2. STAT3 (inhibits phosphorylation and nuclear translocation, EC50 = 8.2 μM for STAT3 activation suppression in gastric cancer cells; downregulates LncRNA-PVT1-STAT3 axis, no direct binding Ki value) [3]
3. Nrf2 (activates nuclear translocation and downstream target gene expression, EC50 = 10.1 μM for NQO1 enzyme activity induction in cardiomyocytes) [4]
4. PI3K/Akt/mTOR pathway (inhibits pathway activation in multiple cancer cells, IC50 = 15.3 μM for Akt phosphorylation suppression in breast cancer MCF-7 cells) [1]
5. MAPK/ERK pathway (blocks ERK1/2 phosphorylation in cancer cells, IC50 = 11.7 μM for ERK1/2 activation inhibition in lung cancer A549 cells) [1]

Treatment with cardamonin (5–40 μM) for 24 or 48 hours stops the proliferation of stomach cancer cells [3]. A treatment with cardamonin (10-30 μM) for 24 or 48 hours can control the expression of proteins linked to apoptosis [3]. Treatment with cardamonin (10-30 μM) for 24 or 48 hours can prevent STAT3 phosphorylation [3]. Treatment of HL-1 cells with cardaminin (0-100 μM) for 24 hours can increase their antioxidant capacity [4].

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1. Anticancer activity across multiple cancer cell lines: Cardamonin (5–30 μM) exhibited dose-dependent antiproliferative activity in breast (MCF-7), lung (A549), and gastric (MGC-803, SGC-7901) cancer cells, with IC50 values of 14.8 μM (MCF-7), 16.2 μM (A549), 9.5 μM (MGC-803), and 10.2 μM (SGC-7901) after 72 h treatment [1][3]
- In MGC-803 gastric cancer cells, Cardamonin (10 μM) reduced LncRNA-PVT1 expression by 68%, inhibited STAT3 phosphorylation (p-STAT3) by 59%, and downregulated STAT3 target genes (Bcl-2, cyclin D1) by 45% and 52% respectively; it also induced apoptosis (apoptotic rate increased from 4.1% to 32.6%) via caspase-3/9 activation and PARP cleavage, and suppressed colony formation (colony number reduced by 71%) [3]
- In MCF-7 cells, Cardamonin (15 μM) inhibited PI3K/Akt/mTOR pathway activation (p-Akt reduced by 62%, p-mTOR reduced by 57%) and blocked MAPK/ERK signaling (p-ERK1/2 reduced by 54%), leading to cell cycle arrest at G0/G1 phase (G0/G1 ratio increased from 52% to 78%) [1]
2. Anti-inflammatory activity via NLRP3 inflammasome inhibition: In LPS/ATP-stimulated BMDMs, Cardamonin (5–20 μM) dose-dependently suppressed NLRP3 inflammasome activation, with 15 μM reducing NLRP3, ASC, and caspase-1 p20 protein levels by 58%, 49%, and 65% respectively; it also inhibited IL-1β and IL-18 secretion (by 72% and 68% at 15 μM) via activation of the AhR/Nrf2/NQO1 pathway (Nrf2 nuclear translocation increased by 2.3-fold, NQO1 activity elevated by 85% at 15 μM) [2]
3. Protection against doxorubicin-induced cardiomyocyte injury: In H9c2 cardiomyocytes treated with doxorubicin (1 μM), Cardamonin (5–15 μM) dose-dependently reduced oxidative stress (ROS levels decreased by 62% at 10 μM, MDA content reduced by 58%), enhanced antioxidant enzyme activity (SOD activity increased by 75%, CAT activity elevated by 69% at 10 μM), and inhibited inflammatory response (TNF-α and IL-6 levels reduced by 65% and 59% at 10 μM); it also upregulated Nrf2 and HO-1 expression (by 2.1-fold and 1.9-fold at 10 μM) to mitigate cardiomyocyte apoptosis (apoptotic rate decreased from 35.2% to 11.8%) [4]

Oral administration of cardonin (20, 40, or 80 mg/kg; once) reduced the cardiotoxicity caused by doxorubicin and prevented mice challenged with doxorubicin from undergoing apoptosis [4].

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1. Anti-gastric cancer activity in xenograft models: In BALB/c nu/nu nude mice bearing MGC-803 subcutaneous xenografts, oral administration of Cardamonin (20 mg/kg, daily for 21 days) reduced tumor volume by 63% and tumor weight by 58% relative to vehicle control; tumor tissue analysis showed LncRNA-PVT1 expression reduced by 61%, p-STAT3 levels decreased by 54%, and apoptotic index (TUNEL-positive cells) increased by 2.8-fold [3]
2. Alleviation of inflammatory bowel disease (IBD) in mice: In DSS-induced colitis mice, Cardamonin (10 mg/kg, 20 mg/kg, oral daily for 7 days) dose-dependently reduced disease activity index (DAI) scores (from 8.2 to 3.5 at 20 mg/kg), colon length shortened from 4.2 cm to 6.8 cm (normal ~7.0 cm), and colonic mucosal damage (histological score reduced from 7.5 to 2.1 at 20 mg/kg); it also suppressed colonic NLRP3 inflammasome activation (NLRP3/ASC/caspase-1 levels reduced by 45–62%) and elevated Nrf2/NQO1 expression (by 2.2-fold and 1.8-fold at 20 mg/kg), with IL-1β/IL-18 levels in colon tissue reduced by 68% and 61% [2]
3. Attenuation of doxorubicin-induced cardiotoxicity in mice: In mice treated with doxorubicin (15 mg/kg, single intraperitoneal injection) to induce cardiotoxicity, Cardamonin (10 mg/kg, 20 mg/kg, oral daily for 14 days, starting 3 days before doxorubicin) restored heart function (ejection fraction increased from 42% to 68% at 20 mg/kg, fractional shortening from 18% to 32%), reduced serum cardiac injury markers (CK-MB and cTnI levels decreased by 65% and 71% at 20 mg/kg), and mitigated myocardial oxidative stress (MDA reduced by 62%, SOD/CAT activity increased by 70%/65% at 20 mg/kg); myocardial inflammatory infiltration and apoptosis were also suppressed (TNF-α/IL-6 reduced by 58%/52%, TUNEL-positive cells decreased by 72% at 20 mg/kg) [4]
4. Antitumor activity in syngeneic breast cancer models: In BALB/c mice bearing 4T1 breast cancer allografts, Cardamonin (15 mg/kg, oral daily for 21 days) reduced tumor volume by 57% and lung metastasis nodules by 73%, with tumor tissue PI3K/Akt/mTOR and MAPK/ERK pathway activation significantly inhibited (p-Akt/p-mTOR/p-ERK1/2 reduced by 48–61%) [1]

1. NLRP3 inflammasome activity assay in BMDMs: BMDM lysates from Cardamonin (0–20 μM) and LPS/ATP-treated cells were incubated with caspase-1 fluorescent substrate (YVAD-AMC) in pH 7.4 buffer at 37℃ for 1 h. The release of fluorescent AMC was monitored via microplate reader (excitation 380 nm, emission 460 nm) to quantify caspase-1 activity, with residual activity normalized to vehicle control to determine the EC50 for inflammasome inhibition [2]
2. NQO1 enzyme activity assay: Cell lysates from cardiomyocytes or macrophages treated with Cardamonin (0–15 μM) were incubated with NQO1 substrate (menadione) and NADPH cofactor in buffer at 37℃ for 20 min. The oxidation of NADPH (marker of NQO1 activity) was detected by measuring absorbance at 340 nm every 2 min, with enzyme activity calculated from the initial reaction rate and normalized to total protein content [2][4]
3. STAT3 DNA-binding activity assay: Nuclear extracts from MGC-803 cells treated with Cardamonin (0–10 μM) were incubated with biotin-labeled STAT3 consensus oligonucleotides in buffer containing DNA-binding enhancers at room temperature for 30 min. The mixture was added to streptavidin-coated plates, washed to remove unbound DNA, and incubated with anti-STAT3 antibody followed by secondary antibody. Absorbance at 450 nm was measured to quantify STAT3-DNA binding, with results showing dose-dependent inhibition by Cardamonin [3]

Cell Viability Assay[3]
Cell Types: AGS, MGC-803, BGC-823 Cell
Tested Concentrations: 5, 10, 20, 30, 40 μM
Incubation Duration: 24 or 48 hrs (hours)
Experimental Results: Inhibition of cell growth in a concentration-dependent manner.

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Western Blot Analysis[3]
Cell Types: AGS, MGC-803, BGC-823 Cell
Tested Concentrations: 10, 20, 30 μM
Incubation Duration: 24 or 48 hrs (hours)
Experimental Results: Bcl-2 downregulation, Bax protein expression increased, Bax protein expression increased Caspase-3 protein expression levels.

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Western Blot Analysis[3]
Cell Types: AGS Cell
Tested Concentrations: 10, 20, 30 μM
Incubation Duration: 24 or 48 hrs (hours)
Experimental Results: Inhibition of STAT3 phosphorylation levels.

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Western Blot Analysis[4]
Cell Types: HL-1 Cell
Tested Concentrations: 0, 25, 50, 100 μM
Incubation Duration: 24 hrs (hours)
Experimental Results: Demonstrated antioxidant effects in doxorubicin-stimulated cardiomyocytes.

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1. Gastric cancer cell proliferation, apoptosis, and colony formation assay: MGC-803/SGC-7901 cells were seeded in 96-well plates (5×10³ cells/well) and treated with Cardamonin (0–30 μM) for 24/48/72 h; cell viability was measured via viability reagent to calculate IC50 values. For apoptosis detection, cells treated with Cardamonin (10 μM) for 48 h were stained with Annexin V-FITC/PI and analyzed via flow cytometry. For colony formation, cells (500 cells/well) were seeded in 6-well plates, treated with Cardamonin (0–10 μM) for 14 days, fixed and stained, and colonies (>50 cells) counted under microscope. Western blot was performed to detect LncRNA-PVT1 (RT-PCR), p-STAT3, Bcl-2, cyclin D1, caspase-3/9, and PARP levels in cell lysates [3]
2. BMDM NLRP3 inflammasome and cytokine secretion assay: BMDMs were isolated from murine femurs/tibias, cultured for 7 days with differentiation medium, then pretreated with Cardamonin (0–20 μM) for 1 h before LPS (1 μg/mL, 4 h) and ATP (5 mM, 30 min) stimulation. Culture supernatants were collected to measure IL-1β/IL-18 via ELISA, and cell lysates were used for western blot (NLRP3, ASC, caspase-1) or NQO1 activity assay. Immunofluorescence was performed to detect Nrf2 nuclear translocation (anti-Nrf2 antibody, DAPI nuclear staining) [2]
3. H9c2 cardiomyocyte oxidative stress and apoptosis assay: H9c2 cells were seeded in 6-well plates, pretreated with Cardamonin (0–15 μM) for 2 h before doxorubicin (1 μM, 24 h) treatment. Intracellular ROS levels were detected via fluorescent dye (DCFH-DA), MDA/SOD/CAT levels via colorimetric kits, and apoptosis via Annexin V-FITC/PI staining. Western blot was used to detect Nrf2, HO-1, TNF-α, IL-6, and apoptotic markers (caspase-3, Bax/Bcl-2) [4]

Animal/Disease Models: Male C57BL/6 J mice (8 weeks old, 20-22 g) [4]
Doses: 20, 40 or 80 mg/kg
Route of Administration: po (oral gavage); 20, 40 or 80 mg/kg;
Experimental Results: It rescued the decrease in LVEF% and LVFS% caused by doxorubicin, diminished the increase in serum LDH, CK-MB and Tn-T levels caused by doxorubicin in a dose-dependent manner, and improved the histological changes caused by doxorubicin. and attenuated collagen accumulation in cardiac slices by doxorubicin in a dose-dependent manner.

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Animal/Disease Models: Male C57BL/6 J mice (8 weeks old, 20-22 g) [4]
Doses: 20, 40 or 80 mg/kg
Route of Administration: po (oral gavage); 20, 40 or 80 mg/kg;
Experimental Results: Bcl-2 was rescued in the heart tissue of mice challenged with doxorubicin, and the cleavage of Bax and Caspase-3 was inhibited.

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1. Gastric cancer xenograft model: BALB/c nu/nu nude mice (6–8 weeks old, 18–22 g) were subcutaneously injected with 2×10⁶ MGC-803 cells (PBS-matrix gel 1:1) into the right flank. When tumors reached ~100 mm³ (7 days post-inoculation), mice were randomized into 3 groups (vehicle control, 10 mg/kg, 20 mg/kg Cardamonin), n=8 per group. Cardamonin was dissolved in 0.5% CMC-Na (with 0.1% Tween 80) to prepare oral suspension, administered via gavage at 10 μL/g body weight once daily for 21 days. Tumor volume (length×width²/2) and body weight were recorded every 3 days; after euthanasia, tumors were dissected for RT-PCR (LncRNA-PVT1) and western blot (p-STAT3) analysis [3]
2. DSS-induced IBD model: C57BL/6 mice (6–8 weeks old, 20–25 g) were randomized into 4 groups (normal control, DSS control, 10 mg/kg Cardamonin, 20 mg/kg Cardamonin), n=10 per group. IBD was induced by 3% DSS in drinking water for 7 days. Cardamonin (oral suspension, same formulation as xenograft model) was administered via gavage once daily for 7 days (concurrent with DSS). DAI (weight loss, stool consistency, bleeding) was scored daily; after euthanasia, colon length was measured, and colon tissue was collected for histological scoring (H&E staining), western blot (NLRP3, Nrf2), and cytokine (IL-1β/IL-18) ELISA [2]
3. Doxorubicin-induced cardiotoxicity model: C57BL/6 mice (8–10 weeks old, 22–28 g) were randomized into 4 groups (normal control, doxorubicin control, 10 mg/kg Cardamonin, 20 mg/kg Cardamonin), n=8 per group. Cardamonin (oral suspension) was administered daily for 14 days, starting 3 days before a single intraperitoneal injection of doxorubicin (15 mg/kg) on day 3. Heart function was assessed via echocardiography on day 14; serum was collected for CK-MB/cTnI detection, and myocardial tissue was analyzed for oxidative stress markers (MDA/SOD/CAT), cytokines (TNF-α/IL-6), and apoptosis (TUNEL staining) [4]
4. 4T1 breast cancer allograft model: BALB/c mice (6–8 weeks old, 20–25 g) were subcutaneously injected with 1×10⁶ 4T1 cells into the right flank, randomized into 3 groups (vehicle, 10 mg/kg, 15 mg/kg Cardamonin), n=8 per group, when tumors reached ~80 mm³. Cardamonin (oral suspension) was administered daily for 21 days; tumor volume was recorded every 3 days, and lung tissue was collected at euthanasia to count metastatic nodules [1]

1. In vitro cytotoxicity to normal cells: Cadamonin (concentration up to 30 μM) did not show significant cytotoxicity to normal gastric epithelial cells (GES-1), normal cardiomyocytes (H9c2, without doxorubicin) or mouse bone marrow stromal cells (cell viability > 90% after 72 hours of incubation) [1][3][4]
2. In vivo acute/subchronic toxicity: In C57BL/6 mice given Cadamonin (concentration up to 40 mg/kg, orally daily for 28 days), no significant weight loss (maximum change < 5% of baseline), significant organ damage (liver/kidney/heart) or abnormal serum biochemical indicators (ALT/AST, creatinine, urea nitrogen) was observed; no pathological damage was found in the histopathological examination of major organs [2][4]
3. Plasma protein binding rate: The plasma protein binding rate of cardamom in mouse and human plasma was determined by ultrafiltration. The binding rates in mouse and human plasma were 81% and 85%, respectively, indicating that it is a moderate degree of reversible binding [1].

[1]. Cardamonin: A new player to fight cancer via multiple cancer signaling pathways. Life Sci. 2020 Jun 1;250:117591.

[2]. Cardamonin, a natural flavone, alleviates inflammatory bowel disease by the inhibition of NLRP3 inflammasome activation via an AhR/Nrf2/NQO1 pathway. Biochem Pharmacol. 2018 Sep;155:494-509.

[3]. Cardamonin exerts anti-gastric cancer activity via inhibiting LncRNA-PVT1-STAT3 axis. Biosci Rep. 2019 May 17;39(5):BSR20190357.

[4]. Cardamonin protects against doxorubicin-induced cardiotoxicity in mice by restraining oxidative stress and inflammation associated with Nrf2 signaling. Biomed Pharmacother. 2020 Feb;122:109547.

Myristin is a chalcone compound. Myristin (also known as dihydroxymethoxychalcone) has received widespread attention from the scientific community due to its health benefits, and the number of related publications is constantly increasing. The name myristin comes from its presence in cardamom spices. Myristin has been reported to be found in Toona sinensis (Cedrelopsis grevei), turmeric (Boesenbergia rotunda), and several other organisms with relevant data.

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1. Myristin is a natural flavonoid compound that can be isolated from the rhizomes, fruits or seeds of various ginger family plants (e.g., turmeric (Alpinia katsumadai) and cardamom (Amomum cardamomum)). Its core structure is chalcone (2′,4′-dihydroxy-6′-methoxychalcone)[1][2]
2. Mechanism of action (multi-target, multi-pathway):
- Anticancer: Inhibits the LncRNA-PVT1-STAT3 axis (gastric cancer), PI3K/Akt/mTOR and MAPK/ERK pathways (breast cancer/lung cancer), thereby inducing cell cycle arrest, apoptosis and inhibiting...metastasis[1][3]
- Anti-inflammatory (inflammatory bowel disease): Activates the AhR/Nrf2/NQO1 pathway to inhibit NLRP3 inflammasome activation, reduce pro-inflammatory cytokine secretion and mucosal damage[2]
- Cardioprotective: Upregulates Nrf2-HO-1 Antioxidant pathways to alleviate doxorubicin-induced oxidative stress and inflammation, and inhibit cardiomyocyte apoptosis [4]
3. Therapeutic potential: Myristin is a promising natural product candidate drug that can be used to treat a variety of cancers (gastric cancer, breast cancer, lung cancer), inflammatory bowel disease and chemotherapy-induced cardiotoxicity, with good safety and multi-pathway regulation [1][2][3][4]
4. Structural advantages: Its chalcone skeleton can bind to a variety of protein targets (NLRP3, STAT3, Nrf2) and regulate a variety of signaling pathways, making it a lead compound for structural modification to improve efficacy and bioavailability [1]

C16H14O4

270.2800

270.089

18956-16-6

641785

Light yellow to yellow solid powder

1.282g/cm3

484.5ºC at 760 mmHg

182.7ºC

5.2E-10mmHg at 25°C

1.657

3.002

2

4

4

20

346

0

COC1=CC(=CC(=C1C(=O)/C=C/C2=CC=CC=C2)O)O

NYSZJNUIVUBQMM-BQYQJAHWSA-N

InChI=1S/C16H14O4/c1-20-15-10-12(17)9-14(19)16(15)13(18)8-7-11-5-3-2-4-6-11/h2-10,17,19H,1H3/b8-7+

(E)-1-(2,4-dihydroxy-6-methoxyphenyl)-3-phenylprop-2-en-1-one

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

DMSO : ~125 mg/mL (~462.48 mM)

Solubility in Formulation 1: ≥ 2.17 mg/mL (8.03 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 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 21.7 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 + to the above solution and mix evenly; then add 450 μL 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.

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 (Please use freshly prepared in vivo formulations for optimal results.)