| Size | Price | |
|---|---|---|
| Other Sizes |
| ln Vitro |
2-Hydroxychalcone prevents triple-negative breast cancer cells from invasively proliferating [1]. 2-Hydroxychalcone suppresses the expression of ICAM-1, VCAM-1, and E-selectin in a concentration-dependent manner, preventing peripheral neutrophils from adhering to the endothelial cell monolayer [2].
In MDA-MB-231 triple negative breast cancer (TNBC) cells, treatment with 2-Hydroxychalcone for 24 h inhibited cell growth with an IC50 of 4.6 μM (95% confidence interval not provided). In Hs578T TNBC cells, the IC50 was 8.41 μM [1]. 2-Hydroxychalcone (20 μM, 4 h treatment) induced apoptosis in MDA-MB-231 cells, as measured by flow cytometry with annexin V-FITC and propidium iodide staining. The percentage of apoptotic cells (early + late) after 20 μM treatment for 4 h was statistically significantly higher than control (p < 0.05 or 0.01). The effect was dose-dependent (concentrations of 5, 10, 20 μM shown) [1]. 2-Hydroxychalcone (5 μM, 24 h) significantly decreased the expression level of the anti-apoptotic protein Bcl-2 in MDA-MB-231 cells (p < 0.01), as determined by immunoblot analysis. The expression of pro-apoptotic Bax was not affected [1]. 2-Hydroxychalcone inhibited the invasive phenotype of MDA-MB-231 cells in a transwell invasion assay. At 5 μM, invasion was significantly inhibited; at 10 μM, inhibition reached 80.2% (p < 0.01). In Hs578T cells, 10 μM 2-Hydroxychalcone inhibited invasion by 46.4% (p < 0.01) [1]. 2-Hydroxychalcone (10–20 μM, 48 h) inhibited the gelatinolytic activity of MMP-9 (but not MMP-2) in a concentration-dependent manner, as measured by gelatin zymography using conditioned media from MDA-MB-231 cells [1]. 2-Hydroxychalcone (20 μM, 24 h with PMA stimulation) reduced the secreted level of MMP-9 protein, as shown by immunoblot analysis. At 20 μM (1 h treatment), it also decreased MMP-9 mRNA levels, as determined by RT-PCR [1]. |
|---|---|
| Enzyme Assay |
Gelatin zymography was performed to assess MMP-2 and MMP-9 gelatinolytic activity. MDA-MB-231 cells were cultured in serum-free medium containing 2-Hydroxychalcone (10 or 20 μM) for 48 h. Conditioned media were collected, centrifuged, concentrated, and equal amounts of protein were mixed with non-reducing sample buffer, then electrophoresed on 10% polyacrylamide gels containing 1 mg/mL gelatin. After electrophoresis, gels were washed with 2.5% Triton X-100, rinsed with Tris-HCl buffer (pH 7.6) containing CaCl2 and Brij-35, incubated overnight at 37°C, stained with Coomassie Brilliant Blue R-250, and destained. Clear bands indicated gelatinase activity. Relative band intensities were quantified [1].
|
| Cell Assay |
MTT assay: MDA-MB-231 or Hs578T cells were seeded in 96-well plates, treated with various concentrations of 2-Hydroxychalcone (1–100 μM) in serum-free medium for 24 h. MTT solution (0.5 mg/mL) was added and incubated for 4 h. Formazan crystals were dissolved in DMSO, and absorbance was measured at 540 nm. Cell viability was expressed as percentage of untreated control. IC50 values were calculated from dose-response curves [1].
Flow cytometric apoptosis assay: MDA-MB-231 cells (80% confluent) were treated with 2-Hydroxychalcone (5, 10, 20 μM) for 4 h. Cells were harvested, washed, and stained with annexin V-FITC and propidium iodide. After incubation, cells were analyzed by flow cytometry. Apoptotic cells were defined as annexin V-positive (early apoptosis) or annexin V/PI double-positive (late apoptosis/necrosis) [1]. Immunoblot analysis: MDA-MB-231 cells were treated with 2-Hydroxychalcone (5 μM in serum-free medium) for 24 h. Cell lysates were subjected to SDS-PAGE, transferred to membranes, and probed with anti-Bcl-2 and anti-Bax antibodies. Protein bands were detected by enhanced chemiluminescence and quantified [1]. In vitro invasion assay: Transwell units with polycarbonate filters were used. The lower side of the filter was coated with type I collagen, and the upper side with Matrigel. 2-Hydroxychalcone was added to both sides of the filter in serum-free medium. Cells were placed in the upper chamber and incubated for 17 h. Filters were fixed, stained with hematoxylin and eosin, and cells on the upper surface were removed. Invaded cells on the lower side were counted under microscopy (400×). Thirteen fields were counted per filter [1]. RT-PCR: Total RNA was isolated from MDA-MB-231 cells treated with 2-Hydroxychalcone (20 μM) for 1 h. Reverse transcription was performed using oligo-dT and Superscript III. MMP-9 primers (forward: 5'-CACACCACAACATCCACTATTG-3', reverse: 5'-CAGGGTTTCCCATCAGCATT-3') were used for PCR (35 cycles, annealing at 57°C). β-actin served as control. PCR products (515 bp for MMP-9) were analyzed by agarose gel electrophoresis [1]. |
| ADME/Pharmacokinetics |
Metabolism / Metabolites
Known metabolites of 2-hydroxychalcone include 2-hydroxychalcone and 2-OH glucuronide. |
| References |
|
| Additional Infomation |
2-Hydroxychalcone is a chalcone derivative with a hydroxyl group on the B-ring (2-position). It is a biosynthetic precursor of flavonoids and exhibits anticancer activities. In this study, it showed more potent antiproliferative and anti-invasive effects against triple negative breast cancer cells (MDA-MB-231, Hs578T) compared to the parent compound chalcone. The inhibitory effects involved induction of apoptosis via downregulation of Bcl-2, and suppression of MMP-9 expression and activity, leading to reduced cell invasion. The compound was dissolved in DMSO as a 100 mM stock and stored at 4°C; final DMSO concentration in media was below 0.1% [1].
|
| Molecular Formula |
C15H12O2
|
|---|---|
| Molecular Weight |
224.26
|
| Exact Mass |
224.084
|
| CAS # |
644-78-0
|
| PubChem CID |
5367146
|
| Appearance |
Light yellow to yellow solid powder
|
| Density |
1.191g/cm3
|
| Boiling Point |
396.566ºC at 760 mmHg
|
| Melting Point |
144-150ºC(lit.)
|
| Flash Point |
169.35ºC
|
| Index of Refraction |
1.654
|
| LogP |
3.288
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
2
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
17
|
| Complexity |
277
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C1=CC=C(C=C1)C(=O)/C=C/C2=CC=CC=C2O
|
| InChi Key |
UDOOPSJCRMKSGL-ZHACJKMWSA-N
|
| InChi Code |
InChI=1S/C15H12O2/c16-14-9-5-4-8-13(14)10-11-15(17)12-6-2-1-3-7-12/h1-11,16H/b11-10+
|
| Chemical Name |
2-Hydroxychalcone
|
| Synonyms |
2-Hydroxychalcone AI3 00855 AI300855AI3-00855
|
| 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: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 : ~125 mg/mL (~557.41 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (9.28 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 20.8 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. Solubility in Formulation 2: ≥ 2.08 mg/mL (9.28 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 20.8 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.4591 mL | 22.2955 mL | 44.5911 mL | |
| 5 mM | 0.8918 mL | 4.4591 mL | 8.9182 mL | |
| 10 mM | 0.4459 mL | 2.2296 mL | 4.4591 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
