Licochalcone A is a naturally occuring chalconoid/phenol and estrogenic chalcone compound extracted from licorice root (Glycyrrhiza glabra or Glycyrrhiza inflata) with antimalarial, anticancer, antibacterial and antiviral activities. Licochalcone A markedly inhibits the in vitro growth of L. major amastigotes in human MDMs and U937 cells. Licochalcone A shows antibacterial effects against all gram-positive bacteria tested and especially against all Bacillus spp. In CT-26 colon cancer cells, Licochalcone A reduces the cell viability and DNA synthesis.

Physicochemical Properties

Molecular Formula	C21H22O4
Molecular Weight	338.3970
Exact Mass	338.151
Elemental Analysis	C, 74.54; H, 6.55; O, 18.91
CAS #	58749-22-7
Related CAS #	58749-22-7
PubChem CID	5318998
Appearance	Yellow to orange solid powder
Density	1.2±0.1 g/cm3
Boiling Point	532.6±50.0 °C at 760 mmHg
Melting Point	100°
Flash Point	186.9±23.6 °C
Vapour Pressure	0.0±1.5 mmHg at 25°C
Index of Refraction	1.611
LogP	4.95
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	6
Heavy Atom Count	25
Complexity	488
Defined Atom Stereocenter Count	0
SMILES	O([H])C1C([H])=C(C(/C(/[H])=C(\[H])/C(C2C([H])=C([H])C(=C([H])C=2[H])O[H])=O)=C([H])C=1C(C([H])=C([H])[H])(C([H])([H])[H])C([H])([H])[H])OC([H])([H])[H]
InChi Key	KAZSKMJFUPEHHW-DHZHZOJOSA-N
InChi Code	InChI=1S/C21H22O4/c1-5-21(2,3)17-12-15(20(25-4)13-19(17)24)8-11-18(23)14-6-9-16(22)10-7-14/h5-13,22,24H,1H2,2-4H3/b11-8+
Chemical Name	(E)-3-(4-hydroxy-2-methoxy-5-(2-methylbut-3-en-2-yl)phenyl)-1-(4-hydroxyphenyl)prop-2-en-1-one
Synonyms	Licochalcone-A; 58749-22-7; Licochalcone-A; (E)-3-(4-hydroxy-2-methoxy-5-(2-methylbut-3-en-2-yl)phenyl)-1-(4-hydroxyphenyl)prop-2-en-1-one; (E)-3-[4-hydroxy-2-methoxy-5-(2-methylbut-3-en-2-yl)phenyl]-1-(4-hydroxyphenyl)prop-2-en-1-one; CHEMBL139702; JTV5467968; 3-Dimethylallyl-4,4'-dihydroxy-6-methoxychalcone;
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	Natural product; UGTs/UDP-glucuronosyltransferases
ln Vitro	Licochalcone A (LCA) demonstrates substantial inhibitory effects against UGT1A1, 1A3, 1A4, 1A6, 1A7, 1A9, and 2B7 (both IC50 and Ki values lower than 5 μM) [2].
ln Vivo	The aim of this study was to determine whether licochalcone A (LCA) has the potential to serve as a beneficial supplement during cisplatin chemotherapy. We found that the administration of LCA alone significantly inhibited the size of the solid tumours in CT-26 cell-inoculated Balb/c mice, without any detectable induction of nephrotoxicity, hepatotoxicity and oxidative stress. LCA also suppressed cell proliferation by reducing DNA synthesis of CT-26 murine colon cancer cells in a dose-dependent manner. LCA did not affect the therapeutic efficacy of cisplatin. Furthermore, LCA inhibited the cisplatin-induced kidney damage characterized by increases in the serum creatinine and blood urea nitrogen, as well as the cisplatin-induced liver damage characterized by increases in the serum alanine aminotransferase and aspartate aminotransferase. The repeated oral administration of LCA prior to cisplatin treatment exerted a preventive effect on the cisplatin-mediated increases in the serum nitric oxide and the tissue lipid peroxidation levels, and recovered the depleted reduced glutathione levels in the tissues. These results suggest that supplementation with LCA may be beneficial in counteracting the side effects of cisplatin therapy in cancer patients.[3] In mice infected with L. major, licochalcone A (5 mg/kg, i.p.) completely prevents lesion development. In mice infected with L. donovani, licochalcone A (150 mg/kg, p.o.) results in > 65 and 85% reductions of parasite loads in the liver and the spleen, respectively. In CT-26 cell-inoculated Balb/c mice, licochalcone A (1 mg/kg, p.o.) inhibits the tumor growth, and alleviates cisplatin-induced nephrotoxicity and hepatotoxicity.
Enzyme Assay	Determination of lipid peroxidation. [3] The content of MDA, as an index of the extent of lipid peroxidation, was assayed in the form of thiobarbituric acid-reactive substances as previously described [31]. The reaction mixture (4 ml) consisted of 0.2 ml of 8.1% sodium dodecyl sulfate, 1.5 ml of 20% acetic acid (pH 3.5), 1.5 ml of 0.8% thiobarbituric acid, 0.2 ml of the homogenate, and distilled water. The mixture was incubated for 1 hr at 95°, cooled for 5 min with tap water, vigorously mixed with 5 ml of a n-butanol-pyridine (15-1, v/v) mixture, and centrifuged for 10 min at 1200 ×g. The absorbance of the organic layer (upper n-butanol phase) was determined at 532 nm. 1,1,3,3-Tetramethoxypropane was utilized to establish the standard curve, and the final MDA concentration was expressed as nmol MDA per mg protein.[3] Determination of GSH level. [3] To determine GSH content in accordance with the method described by Higach, 0.1 ml of the tissue homogenate was added to an equal volume of 10% trichloroacetic acid solution, then centrifuged for 20 min at 1200 ×g. 0.1 ml of the supernatant was added to 0.5 ml of a 0.2 N H2SO4 solution containing 1 mM NaNO2 and incubated for 5 min at room temperature, followed by the addition of 0.2 ml of a 0.5% sulfamic acid ammonium solution, 1 ml of a 0.4 N HCl solution containing 0.1% HgCl2 and 3% sulfanilamide, and 1 ml of a 0.4 N HCl solution containing 0.1% N-(1-naphthyl)ethylenediamine. Five minutes later, the absorbance was determined at 540 nm. The GSH content was expressed as nmol GSH per mg protein using GSH standard calibration curve.
Cell Assay	Licochalcone A (LCA), a flavonoid isolated from the famous Chinese medicinal herb Glycyrrhiza uralensis Fisch, presents obvious anti-cancer effects. In this study, the anti-cancer effects and potential mechanisms of LCA in non-small cell lung cancer (NSCLC) cells were studied. LCA decreased cell viability, increased lactate dehydrogenase release, and induced apoptosis in a concentration-dependent manner in NSCLC cells while not in human embryonic lung fibroblast cells. The expression of phosphatidylethanolamine-modified microtubule-associated protein light-chain 3 (LC3-II) and formation of GFP-LC3 punta, two autophagic markers, were increased after treatment with LCA. LCA-induced LC3-II expression was increased when combined with chloroquine (CQ), while knock-down of autophagy related protein (ATG) 7 or ATG5 reversed LCA-induced LC3-II expression and GFP-LC3 punta formation, suggesting that LCA induced autophagy in NSCLC cells. Inhibition of autophagy could not reverse the LCA-induced cell viability decrease and apoptosis. In addition, LCA increased the expression of endoplasmic reticulum stress related proteins, such as binding immunoglobulin protein and C/EBP homologous protein (CHOP). Knock-down of CHOP reversed LCA-induced cell viability decrease, apoptosis, and autophagy. Taken together, LCA-induced autophagic effect is an accompanied phenomenon in NSCLC cells, and CHOP is critical for LCA-induced cell viability decrease, apoptosis, and autophagy.[1] Human erythrocytes drawn from healthy individuals were exposed for 24 hours to 1-10 µg/ml licochalcone A. Flow cytometry was subsequently employed to estimate phosphatidylserine exposure at the cell surface from annexin V binding, cell volume from forward scatter, [Ca2+]i from Fluo3-fluorescence, and ceramide utilizing specific antibodies. In addition, hemolysis was quantified from hemoglobin release.[2]
Animal Protocol	Mouse xenograft model. In an effort to assess the inhibitory effect of licochalcone A (LCA) on tumour growth, as well as its protective effects against cisplatin-induced nephrotoxicity and hepatotoxicity, the mice were divided into five groups, each group consisting of eight mice: PBS-treated group, CT-26 cell-inoculated group, CT-26 cell-inoculated group with cisplatin, CT-26 cell-inoculated group with LCA, CT-26 cell-inoculated group with LCA and cisplatin. The CT-26 mouse colon cancer cells (2 × 106 cells in 0.1 ml PBS) cultured in DMEM with 10% FBS were subcutaneously injected into the right flanks of the mice. Twenty-four hours later, Balb/c mice were dosed with LCA (1 mg/kg body weight) in PBS via oral gavage. Two hours after treatment with LCA, cisplatin (5 mg/kg body weight) in PBS was intraperitoneally injected. LCA and cisplatin were administered once a day for 15 days. The control group received PBS rather than LCA and cisplatin. On day 15, tumour size was measured with calipers and tumour volumes were calculated in accordance with the following formula: (length × width2)/2. Sixteen hours after the final cisplatin injection, the mice were killed under anaesthesia. The livers and kidneys of the mice were excised immediately after the blood was collected from each mouse, then homogenized for the following experiments.[3] Dissolved in 20 uL of 99% (v/v) ethanol and suspended in 1% carboxymethyl cellulose (CMC) solution; 5mg/kg; Oral gavage or i.p. injection Mice infected with L. major
References	[1]. Induction of C/EBP homologous protein-mediated apoptosis and autophagy by licochalcone A in non-small cell lung cancer cells. Sci Rep. 2016 May 17;6:26241. [2]. Licochalcone A Induced Suicidal Death of Human Erythrocytes. Cell Physiol Biochem. 2015;37(5):2060-70. [3]. Licochalcone A inhibits the growth of colon carcinoma and attenuates cisplatin-induced toxicity without a loss of chemotherapeutic efficacy in mice. Basic Clin Pharmacol Toxicol . 2008 Jul;103(1):48-54.
Additional Infomation	Licochalcone A is a member of chalcones. Licochalcone a has been reported in Glycyrrhiza uralensis, Euphorbia helioscopia, and other organisms with data available. Licochalcone A is a derivative of the phenol chalconoid, found in and extracted from the roots of Glycyrrhiza species G. glabra and inflata, with potential anti-inflammatory, antibacterial, and anticancer activities. Upon administration, licochalcone A inhibits the phosphatidylinositol-3-kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) signaling pathway and inhibits the activity of c-Jun N-terminal kinase 1 (JNK-1), a member of the mitogen-activated protein kinase (MAPK) family that plays a role in the MAPK-mediated signaling pathway. Inhibition of the PI3K/Akt/mTOR- and MAPK-signaling pathways induces cell cycle arrest and apoptosis, decreases migration and invasion of cancer cells, and inhibits tumor cell proliferation. Licochalcone A also prevents the production of reactive oxygen species (ROS), and reduces oxidative stress through the nuclear factor-erythroid 2-related factor 2 (Nrf2) pathway.

Solubility Data

Solubility (In Vitro)

DMSO: 67 mg/mL (198.0 mM) Water:<1 mg/mL Ethanol:67 mg/mL (198.0 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.39 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 2: 2.08 mg/mL (6.15 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. 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 3: 2.08 mg/mL (6.15 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. 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	2.9551 mL	14.7754 mL	29.5508 mL
	5 mM	0.5910 mL	2.9551 mL	5.9102 mL
	10 mM	0.2955 mL	1.4775 mL	2.9551 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.