Gardenin A is an orally bioavailable, synthetic PMF analog with neurotrophic effects on neurite outgrowth and neuronal differentiation. Gardenin A promotes neurogenesis by activating MAPK/ERK, PKC and PKA signaling pathways in PC12 cells, but does not activate TrkA and CREB signaling pathways. Gardenin A also has sedative, anxiolytic, antidepressant, and anticonvulsant (antiepileptic/antiseizure) effects.

Physicochemical Properties

Molecular Formula	C21H22O9
Molecular Weight	418.39
Exact Mass	418.126
CAS #	21187-73-5
PubChem CID	261859
Appearance	Off-white to light yellow solid powder
Density	1.3g/cm3
Boiling Point	624.2ºC at 760 mmHg
Melting Point	162-163ºC
Flash Point	218.5ºC
Vapour Pressure	3.57E-16mmHg at 25°C
Index of Refraction	1.573
LogP	3.217
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	7
Heavy Atom Count	30
Complexity	610
Defined Atom Stereocenter Count	0
InChi Key	MQBFFYQCZCKSBX-UHFFFAOYSA-N
InChi Code	InChI=1S/C21H22O9/c1-24-13-7-10(8-14(25-2)17(13)26-3)12-9-11(22)15-16(23)19(27-4)21(29-6)20(28-5)18(15)30-12/h7-9,23H,1-6H3
Chemical Name	5-hydroxy-6,7,8-trimethoxy-2-(3,4,5-trimethoxyphenyl)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
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

ln Vitro	Neurite outgrowth is induced in PC12 cells by gardenin A (10–20 μM; 48 hours), and neuronal differentiation and synapse formation marker protein, growth-associated protein–43 (GAP–43), and synaptophysin expression are similarly increased (10 μM; 24 and 48 hours) [1]. Using the inhibitor 2-naphthol AS-E phosphate (KG-501) or CREB siRNA, gardenin A (10 μM; 30-120 min) dramatically increases the phosphorylation of the cyclic AMP response element binding protein (CREB) and CRE-mediated transcription [1]. PKA and PKC activity as well as ERK phosphorylation levels are increased by gardenin A (10 μM; 15-120 minutes) [1].
ln Vivo	For mice, gardenin A (0.1–25 mg/kg; oral; single dosage) exhibits a range of neuropharmacological effects, such as sedative, anxiolytic, antidepressant, and anticonvulsant [2]. Mice treated with geniposide A (1–10 mg/kg; oral; single dose) do not have a reduction in motor coordination, and convulsions occur later [2].
Cell Assay	Cell Viability Assay[1] Cell Types: PC12 cells Tested Concentrations: 2, 5, 10, 20 μM Incubation Duration: 48 hrs (hours) Experimental Results: demonstrated cell growth supporting effect at higher concentrations (10-20 µM) and did not exert detectable 237 cytotoxicity on PC12 cells after 48 h incubation in low serum medium. Western Blot Analysis[1] Cell Types: PC12 cells Tested Concentrations: 10 μM Incubation Duration: 0, 15, 30, 60, and 120 min Experimental Results: Dramatically increased the phosphorylation of ERK1/2 (Thr202/Tyr204) at 15 min, subsequently decreased after 30 min. Increased PKC and PKA activity peaked at 15 min. Increased CREB phosphorylation.
Animal Protocol	Animal/Disease Models: Male balb/c (Bagg ALBino) mouse: (24-32 g)[2] Doses: 1 mg/kg, 10 mg/kg, 25 mg/kg Route of Administration: po (oral gavage); single dose Experimental Results: demonstrated sedative effects at a dose of 25 mg/kg, which increased the duration of sleep but did not alter sleep onset.
References	[1]. Neurotrophic action of 5-hydroxylated polymethoxyflavones: 5-demethylnobiletin and gardenin A stimulate neuritogenesis in PC12 cells. J Agric Food Chem. 2013 Oct 2;61(39):9453-63. [2]. Evaluation of the neuropharmacological effects of Gardenin A in mice. Drug Dev Res. 2020 Aug;81(5):600-608.
Additional Infomation	Gardenin A has been reported in Gardenia resinifera, Tamarix dioica, and Murraya paniculata with data available.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.3901 mL	11.9506 mL	23.9011 mL
	5 mM	0.4780 mL	2.3901 mL	4.7802 mL
	10 mM	0.2390 mL	1.1951 mL	2.3901 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.