(2S)-2'-Methoxykurarinone is a compound extracted from Sophora flavescens root and has anti~inflammatory, antipyretic, antidiabetic and anti-tumor activities. (2S)-2'-Methoxykurarinone inhibits osteoclast formation and bone resorption by downregulating RANKL signaling. (2S)-2'-Methoxykurarinone has cytotoxic activity against human myeloid leukemia HL-60 cells.

Physicochemical Properties

Molecular Formula	C27H32O6
Molecular Weight	452.5394
Exact Mass	452.219
CAS #	270249-38-2
PubChem CID	11982641
Appearance	Light yellow to brown solid powder
Density	1.171±0.06 g/cm3 (20 ºC 760 Torr)
Boiling Point	643.6±55.0 °C at 760 mmHg
Melting Point	112-115 ºC
Flash Point	213.2±25.0 °C
Vapour Pressure	0.0±2.0 mmHg at 25°C
Index of Refraction	1.575
LogP	7.05
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	8
Heavy Atom Count	33
Complexity	713
Defined Atom Stereocenter Count	2
SMILES	CC(=CC[C@H](CC1=C2C(=C(C=C1O)OC)C(=O)C[C@H](O2)C3=C(C=C(C=C3)O)OC)C(=C)C)C
InChi Key	KTAQQSUPNZAWEY-OSPHWJPCSA-N
InChi Code	InChI=1S/C27H32O6/c1-15(2)7-8-17(16(3)4)11-20-21(29)13-25(32-6)26-22(30)14-24(33-27(20)26)19-10-9-18(28)12-23(19)31-5/h7,9-10,12-13,17,24,28-29H,3,8,11,14H2,1-2,4-6H3/t17-,24+/m1/s1
Chemical Name	(2S)-7-hydroxy-2-(4-hydroxy-2-methoxyphenyl)-5-methoxy-8-[(2R)-5-methyl-2-prop-1-en-2-ylhex-4-enyl]-2,3-dihydrochromen-4-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

Targets	- RANKL (Receptor Activator of Nuclear Factor-κB Ligand) signaling pathway [1] - NF-κB (Nuclear Factor-κB) signaling pathway [1] - MAPK (Mitogen-Activated Protein Kinase) pathway (ERK1/2, p38, JNK) [1] - Human tumor cell lines (HCT-116: IC50 = 12.5 μM; A549: IC50 = 25.0 μM) [2]
ln Vitro	- (2S)-2'-Methoxykurarinone inhibits osteoclastogenesis and bone resorption via downregulating RANKL signaling. At concentrations of 1, 5, 10 μM, it dose-dependently suppressed RANKL-induced osteoclast formation from RAW264.7 cells by 28±3%, 56±4%, and 78±5%, respectively [1] - It reduced bone resorption activity: 10 μM decreased the area of resorption pits on bone slices by 65±4% [1] - It downregulated RANKL-induced signaling molecules: 10 μM reduced NF-κB p65 nuclear translocation by 62±3%, and phosphorylation of ERK1/2, p38, JNK by 58±4%, 55±3%, 51±2%, respectively [1] - It inhibited the expression of osteoclast-specific genes: 10 μM decreased TRAP, cathepsin K, MMP-9 mRNA levels by 68±5%, 72±4%, 65±3%, and protein levels by 63±4%, 67±3%, 60±2%, respectively [1] - It exhibited cytotoxicity against human tumor cells: IC50 values were 12.5 μM for HCT-116 (colon cancer) and 25.0 μM for A549 (lung cancer) [2]
Enzyme Assay	- TRAP (Tartrate-Resistant Acid Phosphatase) activity assay: RANKL-induced RAW264.7 cells were treated with (2S)-2'-Methoxykurarinone (1, 5, 10 μM) for 5 days. Cells were lysed, and TRAP activity was measured by colorimetric assay using a specific substrate, with absorbance detected at 405 nm [1] - NF-κB activation assay: RAW264.7 cells were pretreated with the compound (10 μM) for 1 hour, then stimulated with RANKL for 30 minutes. Nuclear extracts were prepared, and NF-κB p65 DNA-binding activity was quantified by electrophoretic mobility shift assay (EMSA) [1]
Cell Assay	- Osteoclastogenesis assay: RAW264.7 cells were seeded in 96-well plates (5×10³ cells/well) and treated with RANKL (50 ng/mL) plus (2S)-2'-Methoxykurarinone (1, 5, 10 μM) for 5 days. TRAP staining was performed to count osteoclasts (multinucleated cells with ≥3 nuclei) [1] - Bone resorption assay: Osteoclasts induced from RAW264.7 cells were seeded on bone slices and treated with the compound (10 μM) for 7 days. Resorption pits were visualized by toluidine blue staining and quantified using image analysis software [1] - Western blot/PCR assay: RAW264.7 cells were treated with the compound (1, 5, 10 μM) and RANKL for 24 hours. Total protein was extracted for Western blot (NF-κB p65, p-ERK1/2, p-p38, p-JNK, TRAP, cathepsin K, MMP-9); total RNA was extracted for RT-PCR to detect target gene mRNA levels [1] - Cytotoxicity assay: HCT-116 and A549 cells were seeded in 96-well plates (5×10³ cells/well) and treated with (2S)-2'-Methoxykurarinone (0.78–50 μM) for 72 hours. Cell viability was measured by MTT assay to calculate IC50 values [2]
Toxicity/Toxicokinetics	- No significant cytotoxicity to normal cells: At concentrations up to 10 μM, no obvious viability reduction was observed in primary mouse osteoblasts [1] - Selective cytotoxicity to tumor cells: Lower IC50 for HCT-116 (12.5 μM) compared to A549 (25.0 μM) [2]
References	[1]. (2S)-2'-Methoxykurarinone inhibits osteoclastogenesis and bone resorption through down-regulation of RANKL signaling. Biol Pharm Bull. 2014;37(2):255-61. [2]. Cytotoxic lavandulyl flavanones from Sophora flavescens. J Nat Prod. 2000 May;63(5):680-1.
Additional Infomation	(2S)-2'-methoxykurarinone is a dimethoxyflavanone that is (2S)-(-)-kurarinone in which the hydroxy group at position 2' is replaced by a methoxy group. Isolated from the roots of Sophora flavescens, it exhibits cytotoxicity against human myeloid leukemia HL-60 cells. It has a role as a metabolite and an antineoplastic agent. It is a dimethoxyflavanone, a dihydroxyflavanone and a member of 4'-hydroxyflavanones. It is functionally related to a (2S)-(-)-kurarinone. (2S)-2'-methoxykurarinone has been reported in Sophora flavescens with data available. - (2S)-2'-Methoxykurarinone is a lavandulyl flavanone isolated from the roots of Sophora flavescens Ait. [2] - Its anti-osteoclastogenic mechanism involves blocking RANKL-mediated activation of NF-κB and MAPK (ERK1/2, p38, JNK) pathways, thereby suppressing osteoclast differentiation and function [1] - It exhibits dual biological activities: inhibiting osteoclastogenesis (potential for osteoporosis treatment) and exerting cytotoxicity against tumor cells [1][2]

Solubility Data

Solubility (In Vitro)

DMSO : ~25 mg/mL (~55.24 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.52 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 25.0 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: 2.5 mg/mL (5.52 mM) 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; with ultrasonication. 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 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	2.2097 mL	11.0487 mL	22.0975 mL
	5 mM	0.4419 mL	2.2097 mL	4.4195 mL
	10 mM	0.2210 mL	1.1049 mL	2.2097 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.