INCA-6 (Triptycene-1,4-quinone) is a cell-permeable (penetrable) NFAT inhibitor. INCA-6 specifically blocks the targeting of NFAT substrates to the calcineurin site and is a potent inhibitor of calcineurin (CN)-NFAT signaling.

Physicochemical Properties

Molecular Formula	C20H12O2
Molecular Weight	284.31
Exact Mass	284.084
CAS #	3519-82-2
PubChem CID	230748
Appearance	Light yellow to yellow solid powder
Density	1.38g/cm3
Boiling Point	464.2ºC at 760 mmHg
Flash Point	172.6ºC
Index of Refraction	1.729
LogP	3.282
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	2
Rotatable Bond Count	0
Heavy Atom Count	22
Complexity	535
Defined Atom Stereocenter Count	0
InChi Key	GCHPUOHXXCNSQL-UHFFFAOYSA-N
InChi Code	InChI=1S/C20H12O2/c21-15-9-10-16(22)20-18-12-6-2-1-5-11(12)17(19(15)20)13-7-3-4-8-14(13)18/h1-10,17-18H
Chemical Name	pentacyclo[6.6.6.02,7.09,14.015,20]icosa-2(7),4,9,11,13,15,17,19-octaene-3,6-dione
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	In 3-Hz cells, INCA-6 (5 μM; for 24 hours) inhibits the transitory downregulation of outward K+ current (Ito)[1]. When BV-2 cells are pre-treated with INCA-6 (10 μM), ATP-induced CXCL2 production in BV-2 cells is considerably inhibited. Moreover, ATP-induced CXCL2 production in rat primary microglia is inhibited by INCA-6[2]. TIPASIGRGIN (TG) is not necessary for the reduction of SERCA2 transcript levels or protein expression caused by INCA-6 (5 μM)[3]. Human retinal microvascular endothelial cells (HRMECs) stimulated by serum and VEGF are considerably inhibited by INCA-6 (1.0 and 2.5 μM; 24 hours) treatment, but baseline proliferation is unaffected[4].
ln Vivo	In oxygen-induced retinopathy (OIR), INCA-6 (5.0, or 25.0 μM) therapy effectively lowers pathologic neovascularization[4].
Cell Assay	Cell Proliferation Assay[4] Cell Types: Human retinal microvascular endothelial cells Tested Concentrations: 0.5, 1.0, or 2.5 μM Incubation Duration: 24 hrs (hours) Experimental Results: Dramatically inhibited VEGF-induced proliferation at 1.0 and 2.5 μM concentrations.
Animal Protocol	Animal/Disease Models: Rats bearing OIR model[4] Doses: 2.5, 5.0, or 25.0 μM Route of Administration: Intravitreal injection on days 14(0) and 14(3) Experimental Results: diminished the severity of OIR in a dose dependent manner. Significant inhibition was seen at 5.0 and 25.0 μM concentrations.
References	[1]. Mechanisms underlying rate-dependent remodeling of transient outward potassium current in canine ventricular myocytes. Circ Res. 2008 Sep 26;103(7):733-42. [2]. P2X7 receptor activation induces CXCL2 production in microglia through NFAT and PKC/MAPK pathways. J Neurochem. 2010 Aug;114(3):810-9. [3]. Silencing calcineurin A subunit reduces SERCA2 expression in cardiac myocytes. Am J Physiol Heart Circ Physiol. 2011 Jan;300(1):H173-80. [4]. The role of the NFAT signaling pathway in retinal neovascularization. Invest Ophthalmol Vis Sci. 2013 Oct 25;54(10):7020-7.
Additional Infomation	LSM-2729 is an anthraquinone.

Solubility Data

Solubility (In Vitro)

DMSO: 2.86 mg/mL (10.06 mM)

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	3.5173 mL	17.5864 mL	35.1729 mL
	5 mM	0.7035 mL	3.5173 mL	7.0346 mL
	10 mM	0.3517 mL	1.7586 mL	3.5173 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.