Puerarin (Kakonein), a naturally occuring isoflavone isolated from the root of Radix puerariae, is a 5-HT2C receptor and benzodiazepine site antagonist. Puerarin (Kakonein), an isoflavone that binds to the benzodiazepine site and the 5-HT2C receptor is present in the root of Radix puerariae. In China, pulmonary ailments are treated clinically with pulmonaria. Puerarin has been shown in another study to have anti-cancer qualities. Puerarin, at a dose-dependent 25 μM, inhibits the growth of HT-29 cells by increasing bax and decreasing c-myc and bcl-2.

Physicochemical Properties

Molecular Formula	C21H20O9
Molecular Weight	416.3781
Exact Mass	416.11
Elemental Analysis	C, 60.58; H, 4.84; O, 34.58
CAS #	3681-99-0
Related CAS #	3681-99-0
PubChem CID	5281807
Appearance	White to off-white solid powder
Density	1.6±0.1 g/cm3
Boiling Point	791.2±60.0 °C at 760 mmHg
Melting Point	187-189°C
Flash Point	281.5±26.4 °C
Vapour Pressure	0.0±2.9 mmHg at 25°C
Index of Refraction	1.717
LogP	-0.67
Hydrogen Bond Donor Count	6
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	3
Heavy Atom Count	30
Complexity	659
Defined Atom Stereocenter Count	5
SMILES	C1=CC(=CC=C1C2=COC3=C(C2=O)C=CC(=C3[C@H]4[C@@H]([C@H]([C@@H]([C@H](O4)CO)O)O)O)O)O
InChi Key	HKEAFJYKMMKDOR-VPRICQMDSA-N
InChi Code	InChI=1S/C21H20O9/c22-7-14-17(26)18(27)19(28)21(30-14)15-13(24)6-5-11-16(25)12(8-29-20(11)15)9-1-3-10(23)4-2-9/h1-6,8,14,17-19,21-24,26-28H,7H2/t14-,17-,18+,19-,21+/m1/s1
Chemical Name	7-hydroxy-3-(4-hydroxyphenyl)-8-[(2S,3R,4R,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]chromen-4-one
Synonyms	Kakonein; NPI-031G; NPI031G; NPI 031G; Puerarin
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	5-HT2C Receptor
ln Vitro	In vitro activity: Puerarin (Kakonein), an isoflavone that binds to the benzodiazepine site and the 5-HT2C receptor is present in the root of Radix puerariae.[1] In China, pulmonary ailments are treated clinically with pulmonaria.[2] Puerarin has been shown in another study to have anti-cancer qualities. Puerarin, at a dose-dependent 25 μM, inhibits the growth of HT-29 cells by increasing bax and decreasing c-myc and bcl-2.[3]
ln Vivo	Puerarin (300 mg/kg/day, p.o.) significantly reduced the elevated total cholesterol brought on by the hypercholesterolmic diet in both the serum and the liver of rats given a hypercholesterolmic diet plus Puerarin administration. [2] LD50: 738 mg/kg (i.v.) in mice [4]
Enzyme Assay	Puerarin (7,4'-dihydroxy-8-C-glucosylisoflavone) is the most abundant isoflavone-C-glucoside extracted from Radix puerariae, and it has been used for various medicinal purposes in traditional oriental medicine for thousands of years. In the present study, the ability of the puerarin to modulate inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and C reactive protein (CRP) expression and induce changes in the nuclear factor κB (NF-κB) pathway in RAW264.7 macrophage cells was examined. The protein and mRNA levels of lipopolysaccharide (LPS)-induced iNOS, COX-2 and CRP were determined in RAW246.7 macrophage cells. Inhibitor κB (I-κB) phosphorylation and p65NF-κB expression in RAW246.7 macrophage cells were also detected under our experimental conditions. The results indicated that puerarin inhibited the expression of LPS-induced iNOS, COX-2 and CRP proteins and also suppressed their mRNAs from RT-PCR experiments in RAW264.7 cells. Subsequently, we determined that the inhibition of iNOS, COX-2 and CRP expression was due to a dose-dependent inhibition of phosphorylation and degradation of I-κB, which resulted in the reduction of p65NF-κB nuclear translocation. These data suggested that the effect of puerarin-mediated inhibition of LPS-induced iNOS, COX-2 and CRP expression is attributed to suppressed NF-κB activation at the transcriptional level [4].
Cell Assay	RAW264.7 cells are kept at subconfluence in a humidified environment that is 95% air and 5% CO2 and is kept at 37°C. Dulbecco's Modified Eagle's Medium, enhanced with 10% fetal bovine serum, penicillin (100 units/mL), and streptomycin (100 μg/mL), is the medium utilized for routine subculture. Following puerarin treatment, the cells' viability is assessed using the MTT assay. The cells are incubated at 37°C with MTT (0.05 mg/mL) for 4 hours, after which the supernatants are removed for nitrite determination. The optical density is then measured at 540 nm. Puerarin is present in concentrations of 10, 20, 40, and 100 μM.
Animal Protocol	30Rats: Three groups of seven-week-old, healthy male SD rats are randomly assigned to receive puerarin treatment at three different dosages: high (H), moderate (M), and low (L). Using 0.9% saline as a resuspension medium, purerarin is administered intragastrically to patients in three different groups: L (0.25 mg/(kg×d), M (0.5 mg/(kg×d), and H (1.0 mg/(kg×d)) for eight days in a row. Rats that are model and control are given the same amount of saline at the same time . Mice: A daily cycle of 12 hours light and 12 hours dark, with a room temperature of 22±2 °C and a relative humidity of 55%±5%, is used to acclimate 40 male ICR mice (weight: 20-22 g). Following a week of adaptation, the mice are split into four groups of ten mice each at random. The mice are given 0.1 M equimolar concentrations of genistein and puerarin in a sodium carboxymethyl cellulose solution (gastric volume: 3 mL kg-1 body weight).
References	[1]. Pharmacol Biochem Behav. 2003 Jun;75(3):619-25. [2]. Life Sci. 2006 Jun 20;79(4):324-30. [3]. Cancer Lett. 2006 Jul 8;238(1):53-60. [4]. Pharmacol Rep. 2011;63(3):781-9.
Additional Infomation	Puerarin is a hydroxyisoflavone that is isoflavone substituted by hydroxy groups at positions 7 and 4' and a beta-D-glucopyranosyl residue at position 8 via a C-glycosidic linkage. It has a role as a plant metabolite, an autophagy inducer, a cardioprotective agent, an antioxidant, an anti-inflammatory agent, an antipyretic and a ferroptosis inhibitor. It is a C-glycosyl compound and a hydroxyisoflavone. It is functionally related to an isoflavone. It is a conjugate acid of a puerarin(1-). Puerarin has been investigated for the treatment of Alcohol Abuse. Puerarin has been reported in Bupleurum chinense, Pueraria calycina, and other organisms with data available.

Solubility Data

Solubility (In Vitro)

DMSO: 50~86 mg/mL (120.1~206.5 mM) Water: <1 mg/mL Ethanol: <1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 3 mg/mL (7.20 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 30.0 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: ≥ 3 mg/mL (7.20 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 30.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 3: 3 mg/mL (7.20 mM) in 10% DMSO + 90% Corn Oil (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 30.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 4: 20 mg/mL (48.03 mM) in 20% SBE-β-CD in Saline (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. 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	2.4017 mL	12.0083 mL	24.0165 mL
	5 mM	0.4803 mL	2.4017 mL	4.8033 mL
	10 mM	0.2402 mL	1.2008 mL	2.4017 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.