Gardenin B is a natural flavonoid extracted from Gardenia jasminoides. Gardenin B can induce the death of human leukemia cells, and the induced apoptosis is related to multiple caspases.

Physicochemical Properties

Molecular Formula	C19H18O7
Molecular Weight	358.3420
Exact Mass	358.105
CAS #	2798-20-1
PubChem CID	96539
Appearance	Light yellow to green yellow solid powder
Density	1.3±0.1 g/cm3
Boiling Point	582.0±50.0 °C at 760 mmHg
Melting Point	180-181ºC
Flash Point	210.8±23.6 °C
Vapour Pressure	0.0±1.7 mmHg at 25°C
Index of Refraction	1.593
LogP	2.27
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	5
Heavy Atom Count	26
Complexity	526
Defined Atom Stereocenter Count	0
InChi Key	LXEVSYZNYDZSOB-UHFFFAOYSA-N
InChi Code	InChI=1S/C19H18O7/c1-22-11-7-5-10(6-8-11)13-9-12(20)14-15(21)17(23-2)19(25-4)18(24-3)16(14)26-13/h5-9,21H,1-4H3
Chemical Name	5-hydroxy-6,7,8-trimethoxy-2-(4-methoxyphenyl)chromen-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	- Caspase family (caspase-3, caspase-8, caspase-9) [1] - Apoptosis-related proteins (Bax, Bcl-2) [1] - Human leukemia cells (HL-60: IC50 = 12.5 ± 1.3 μM; U937: IC50 = 15.7 ± 1.5 μM) [1]
ln Vitro	- Gardenin B exhibited dose-dependent cytotoxicity against human leukemia cells. For HL-60 cells, IC50 was 12.5 ± 1.3 μM; for U937 cells, IC50 was 15.7 ± 1.5 μM (48-hour incubation) [1] - It induced apoptosis in leukemia cells: 20 μM Gardenin B increased the apoptotic rate of HL-60 cells by 58±4% and U937 cells by 52±3% (Annexin V-FITC/PI staining) [1] - The compound activated multiple caspases: 20 μM increased cleaved caspase-3, -8, -9 protein levels by 3.2±0.3, 2.8±0.2, and 2.5±0.2-fold, respectively, in HL-60 cells [1] - It altered the Bax/Bcl-2 ratio: 20 μM elevated the Bax/Bcl-2 ratio by 3.8±0.3-fold in HL-60 cells, promoting mitochondrial apoptotic pathway activation [1] - It did not induce reactive oxygen species (ROS) generation: Even at 30 μM, ROS levels in HL-60 cells showed no significant difference from the control group [1] - It showed low cytotoxicity to normal human peripheral blood mononuclear cells (PBMCs): Cell viability remained above 80% at concentrations up to 30 μM [1]
Enzyme Assay	- Caspase activity assay: HL-60 cells were treated with Gardenin B (10, 20, 30 μM) for 24 hours. Cell lysates were incubated with caspase-3/-8/-9 specific fluorogenic substrates, and fluorescence intensity was measured at excitation/emission wavelengths of 400/505 nm to quantify caspase activity [1] - ROS detection assay: Leukemia cells were loaded with a ROS-specific fluorescent probe after treatment with Gardenin B (10–30 μM) for 24 hours. Fluorescence at 488/525 nm was detected to confirm no ROS overproduction [1]
Cell Assay	- Cell viability assay: HL-60 and U937 cells were seeded in 96-well plates (5×10³ cells/well) and incubated overnight. Gardenin B (5–30 μM) was added, and cells were cultured for 48 hours. MTT reagent was added, and absorbance was measured at 570 nm to calculate IC50 values [1] - Apoptosis detection assay: Leukemia cells were treated with Gardenin B (10, 20 μM) for 48 hours, stained with Annexin V-FITC and PI, and analyzed by flow cytometry to determine apoptotic rates [1] - Protein expression assay: Cells were treated with the compound (10, 20 μM) for 24 hours. Total protein was extracted for Western blot analysis of cleaved caspase-3/-8/-9, Bax, and Bcl-2 [1] - Normal cell cytotoxicity assay: Human PBMCs were treated with Gardenin B (10–30 μM) for 48 hours, and cell viability was measured by MTT assay [1]
Toxicity/Toxicokinetics	- Selective toxicity: Potent cytotoxicity to leukemia cells (IC50 12.5–15.7 μM) but low toxicity to normal PBMCs (viability >80% at 30 μM) [1] - No oxidative stress induction: Did not generate ROS in leukemia cells, avoiding non-specific oxidative damage [1]
References	[1]. Gardenin B-induced cell death in human leukemia cells involves multiple caspases but is independent of the generation of reactive oxygen species. Chem Biol Interact. 2016 Aug 25;256:220-7.
Additional Infomation	Gardenin B is a tetramethoxyflavone that is tangeretin in which the methoxy group at position 5 has been replaced by a hydroxy group. It has a role as a plant metabolite. It is a tetramethoxyflavone and a monohydroxyflavone. It is functionally related to a tangeretin. Gardenin B has been reported in Citrus tankan, Citrus reticulata, and other organisms with data available. - Gardenin B is a natural flavonoid compound isolated from plant sources [1] - Its anti-leukemia mechanism involves activating the caspase-dependent apoptotic pathway (intrinsic and extrinsic pathways) by upregulating Bax, downregulating Bcl-2, and cleaving caspases-3/-8/-9 [1] - It induces cell death independent of ROS generation, distinguishing it from ROS-mediated anti-cancer agents [1] - It shows potential as a targeted anti-leukemia candidate due to its selective toxicity to leukemia cells [1]

Solubility Data

Solubility (In Vitro)

DMSO : ~10 mg/mL (~27.91 mM)

Solubility (In Vivo)

Solubility in Formulation 1: 2.5 mg/mL (6.98 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 sonication. 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. Solubility in Formulation 2: 1.67 mg/mL (4.66 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 16.7 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.7906 mL	13.9532 mL	27.9065 mL
	5 mM	0.5581 mL	2.7906 mL	5.5813 mL
	10 mM	0.2791 mL	1.3953 mL	2.7906 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.