Lespenefril is a novel and potent agent of antioxidant, antimicrobial and anti-inflammatory. It possesses a variety of bioactivities, including chemopreventive, antinociception, anti-inflammatory, anti-diabetic, and antitumor effects. Additionally, lepenefril stimulates the insulin signaling pathway.

Physicochemical Properties

Molecular Formula	C27H30O14
Molecular Weight	578.52
Exact Mass	578.163
Elemental Analysis	C, 56.06; H, 5.23; O, 38.72
CAS #	482-38-2
Related CAS #	482-38-2
PubChem CID	5486199
Appearance	Light yellow to yellow solid powder
Density	1.7±0.1 g/cm3
Boiling Point	908.6±65.0 °C at 760 mmHg
Melting Point	202 - 203 °C
Flash Point	302.8±27.8 °C
Vapour Pressure	0.0±0.3 mmHg at 25°C
Index of Refraction	1.725
LogP	1.66
Hydrogen Bond Donor Count	8
Hydrogen Bond Acceptor Count	14
Rotatable Bond Count	5
Heavy Atom Count	41
Complexity	968
Defined Atom Stereocenter Count	10
SMILES	O=C1C(O[C@H](O[C@@H](C)[C@H](O)[C@H]2O)[C@@H]2O)=C(C3=CC=C(O)C=C3)OC4=CC(O[C@H](O[C@@H](C)[C@H](O)[C@H]5O)[C@@H]5O)=CC(O)=C14
InChi Key	PUPKKEQDLNREIM-QNSQPKOQSA-N
InChi Code	InChI=1S/C27H30O14/c1-9-17(30)20(33)22(35)26(37-9)39-13-7-14(29)16-15(8-13)40-24(11-3-5-12(28)6-4-11)25(19(16)32)41-27-23(36)21(34)18(31)10(2)38-27/h3-10,17-18,20-23,26-31,33-36H,1-2H3/t9-,10-,17-,18-,20+,21+,22+,23+,26-,27-/m0/s1
Chemical Name	5-hydroxy-2-(4-hydroxyphenyl)-3,7-bis[[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy]chromen-4-one
Synonyms	BRN 0073958; Kaempferol 3,7-bisrhamnoside; Kaempferitrin
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	Insulin Receptor
ln Vitro	Kaempferitrin activates insulin signaling pathway. At 1–20 μM, kaempferitrin causes over 90% of matured 3T3–L1 adipocytes to survive; at 25 and 50 μM, the survival rates start to rapidly decline. The effects of 10 nM insulin are comparable to those of kaempferitrin (15 μM), which increases the tyrosine phosphorylation of the insulin receptor substrate 1 and the beta tyrosine phosphorylation of the insulin receptor. Additionally, wortmannin, a PI3-K inhibitor, can inhibit the stimulation of akt phosphorylation on ser473 caused by kaempferitrin (15 μM). At 15 μM, kaempferitrin potently induces GLUT4 translocation to the adipocyte membrane; wortmannin inhibits this. Furthermore, in mature 3T3-L1 adipocytes, kaempferitrin increases secreted adiponectin and total levels of Glu4 protein[1]. With IC50s of 45 ± 2.6 and 65 ± 2.6 μM, kaempferitrin is cytotoxic to human cancer cells like HeLa and MDA-MB231 cells, but has minimal toxic effects on non-tumorigenic cells. After treating HeLa cells for 24 and 48 hours, kaempferitrin (45 μM) causes the cells to undergo apoptosis and generate reactive oxygen species (ROS). Additionally, Kaempferitrin (45 μM) causes the expression of proteins linked to the intrinsic pathway of apoptosis, inhibits G1 arrest, and activates caspase 3 in HeLa cells[2].
ln Vivo	Kaempferitrin (2.5, 10 and 25 mg/kg, i.p.) ignificantly reduces tumor weight by 37%, 81%, and 95%, and inhibits tumor growth by 40%, 87%, and 97%, respectively in nu/nu mice bearing HeLa tumor. In mice with tumors, kaempferitrin also reduces cell division and increases longevity[2].
Cell Assay	The MTT assay is used to perform the viability assay. To achieve confluence, preconfluent 3T3-L1 preadipocytes are seeded. After confluence (day 0), kempferitrin is added in place of insulin, and three hours later, the compound's viability is assessed. On day eight, a second cell viability test is conducted. In the case of kaempferitrin plus insulin therapy, different kaempferitrin concentrations are administered concurrently with 0.2 nM insulin from day 0 to day 8. The control cells are those that were not treated with insulin or kaempferitrin from day 0 to day 8 of differentiation. At the conclusion of the 24 and 48-hour compound treatment periods, MTT cells are cultured for 3 hours in order to assess the viability of the matured 3T3-L1 cells treated with insulin or kaempferitrin. The survival rates are then computed and compared to those of the cells treated without insulin or kaempferitrin[1].
Animal Protocol	The nu/nu mice receive a subcutaneous injection of 1.5 × 106 HeLa cells in their backs. Groups of five mice are given doses of either paclitaxel (PCX) at 1 mg/kg or cisplatin (CDDP) at 1 mg/kg intraperitoneally every day for 32 days, or 2.5 to 25 mg/kg of Kaempferitrin dissolved in 0.1 mL of 0.9% saline solution, four hours after the tumor is implanted. 0.1 mL of the vehicle solution was given to the animal control group. Tumor volume is calculated by multiplying length by width by height2. A vernier caliper is used to measure the size of tumors in millimeters. When the trials are over, the animals are slaughtered, and their tumors are removed and weighed[2].
References	[1]. Kaempferitrin activates the insulin signaling pathway and stimulates secretion of adiponectin in 3T3-L1 adipocytes. Eur J Pharmacol. 2009 Apr 1;607(1-3):27-34. [2]. Kaempferitrin induces apoptosis via intrinsic pathway in HeLa cells and exerts antitumor effects. J Ethnopharmacol. 2013 Jan 30;145(2):476-89.
Additional Infomation	Kaempferol 3,7-di-O-alpha-L-rhamnoside is a glycosyloxyflavone that is kaempferol attached to alpha-L-rhamnopyranosyl residues at positions 3 and 7 respectively via glycosidic linkages. It has been isolated from the aerial parts of Vicia faba and Lotus edulis. It has a role as a bone density conservation agent, a hypoglycemic agent, an immunomodulator, an anti-inflammatory agent, an antineoplastic agent, a plant metabolite, an apoptosis inducer and an antidepressant. It is an alpha-L-rhamnoside, a monosaccharide derivative, a dihydroxyflavone, a glycosyloxyflavone and a polyphenol. It is functionally related to a kaempferol. Kaempferitrin has been reported in Camellia sinensis, Annulohypoxylon bovei, and other organisms with data available. See also: Selenicereus grandiflorus stem (part of).

Solubility Data

Solubility (In Vitro)

DMSO: 25~100 mg/mL (43.2~172.9 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (3.60 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 (3.60 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. Solubility in Formulation 3: ≥ 2.08 mg/mL (3.60 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.7285 mL	8.6427 mL	17.2855 mL
	5 mM	0.3457 mL	1.7285 mL	3.4571 mL
	10 mM	0.1729 mL	0.8643 mL	1.7285 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.