Flufenamic Acid (CI-440; CN-27554) is an anti-inflammatory agent of the NSAIDs (non-steroidal anti-inflammatory agent) class. It inhibits cyclooxygenase (COX), activates AMPK, and also modulates ion channels, blocking chloride channels and L-type Ca2+ channels, modulating non-selective cation channels (NSC), activating K+ channels. . Flufenamic acids reversibly inhibits ICl(Ca) in Xenopus oocytes with IC50 of 28 mM, elicit in response to depolarizing voltage steps, in a dose-dependent manner, with no effect on the shape of the current-voltage curve. Flufenamic acids blocks Ca2(+)-activated non-selective cation channels in inside-out patches from the basolateral membrane of rat exocrine pancreatic cells with IC50 of 10 μM.

Physicochemical Properties

Molecular Formula	C14H10F3NO2
Molecular Weight	281.23
Exact Mass	281.066
CAS #	530-78-9
Related CAS #	Flufenamic acid-d4;1185071-99-1;Flufenamic acid-13C6;1325559-30-5
PubChem CID	3371
Appearance	White to off-white solid powder
Density	1.4±0.1 g/cm3
Boiling Point	373.9±42.0 °C at 760 mmHg
Melting Point	132-135 °C(lit.)
Flash Point	179.9±27.9 °C
Vapour Pressure	0.0±0.9 mmHg at 25°C
Index of Refraction	1.585
LogP	5.62
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	3
Heavy Atom Count	20
Complexity	346
Defined Atom Stereocenter Count	0
SMILES	FC(C1C([H])=C([H])C([H])=C(C=1[H])N([H])C1=C([H])C([H])=C([H])C([H])=C1C(=O)O[H])(F)F
InChi Key	LPEPZBJOKDYZAD-UHFFFAOYSA-N
InChi Code	InChI=1S/C14H10F3NO2/c15-14(16,17)9-4-3-5-10(8-9)18-12-7-2-1-6-11(12)13(19)20/h1-8,18H,(H,19,20)
Chemical Name	2-((3-(trifluoromethyl)phenyl)amino)benzoic acid
Synonyms	CI-440; CN-27554; INF-1837; CI 440; CN 27554; INF 1837; CI440; CN27554; INF1837
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	Cyclooxygenase (COX) is inhibited by the nonsteroidal anti-inflammatory medication flufenamic acid. Furthermore, it has the ability to control non-selective cation channels (NSC), block L-type Ca2+ channels, regulate ion channels, and modify chloride channels. K+ channel activation. A minimum of two TRP channels (C6 and A1) are activated by flufenamic acid, while numerous TRP channels (C3, C7, M2, M3, M4, M5, M7, M8, V1, V3, and V4) are inhibited [1]. CaMKKβ, a calcium/calmodulin-dependent protein kinase kinase beta, is directly stimulated by flufenamic acid to cause AMPK activation in T84 cells [2]. Moreover, Flufenamic acid (FFA; 5-50 μM) inhibits Cl- secretion that is dependent on cAMP in intact T84 cells, apical ICl-mediated by CFTR is inhibited, and Ca2+-dependent Cl- secretion is blocked in a dose-dependent manner. At 100 μM, the IC50 of FFA's Cl-secretion is approximately 10 μM, and it almost completely blocks the T84 cell monolayer. However, it has no effect on Na+-K+ ATPase or NKCC in T84 cells [3].
ln Vivo	In a mouse model of Vibrio cholerae El Tor variant (EL)-induced diarrhea, flufenamic acid (50 mg/kg, ip) exhibits anti-inflammatory effects. At 20 mg/kg, it greatly reduces EL-induced intestinal secretion and breakdown of the barrier. Furthermore, in the gut of mice infected with EL, flufenamic acid stimulates AMPK activation and suppresses pro-inflammatory mediator production and NF-κB nuclear translocation [2].
Animal Protocol	50 mg/kg, i.p. Mouse
References	[1]. Guinamard R, et al. Flufenamic acid as an ion channel modulator. Pharmacol Ther. 2013 May;138(2):272-84. [2]. Pongkorpsakol P, et al. Flufenamic acid protects against intestinal fluid secretion and barrier leakage in a mouse model of Vibrio cholerae infection through NF-κB inhibition and AMPK activation. Eur J Pharmacol. 2017 Mar 5;798:94-104. [3]. Pongkorpsakol P, et al. Cellular mechanisms underlying the inhibitory effect of flufenamic acid on chloride secretion in human intestinal epithelial cells. J Pharmacol Sci. 2017 Jun;134(2):93-100. [4]. Pobbati AV, et al. Targeting the Central Pocket in Human Transcription Factor TEAD as a Potential Cancer Therapeutic Strategy. Structure. 2015;23(11):2076-2086
Additional Infomation	Flufenamic acid is an aromatic amino acid consisting of anthranilic acid carrying an N-(trifluoromethyl)phenyl substituent. An analgesic and anti-inflammatory, it is used in rheumatic disorders. It has a role as an EC 1.14.99.1 (prostaglandin-endoperoxide synthase) inhibitor, a non-steroidal anti-inflammatory drug, a non-narcotic analgesic and an antipyretic. It is an aromatic amino acid and an organofluorine compound. It is functionally related to a diphenylamine, an anthranilic acid and a (trifluoromethyl)benzene. It is a conjugate acid of a flufenamate. An anthranilic acid derivative with analgesic, anti-inflammatory, and antipyretic properties. It is used in musculoskeletal and joint disorders and administered by mouth and topically. (From Martindale, The Extra Pharmacopoeia, 30th ed, p16) An anthranilic acid derivative with analgesic, anti-inflammatory, and antipyretic properties. It is used in musculoskeletal and joint disorders and administered by mouth and topically. (From Martindale, The Extra Pharmacopoeia, 30th ed, p16)

Solubility Data

Solubility (In Vitro)

DMSO:56 mg/mL (199.1 mM) Water:<1 mg/mL Ethanol:56 mg/mL (199.1 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (8.89 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 25.0 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: ≥ 2.5 mg/mL (8.89 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 25.0 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	3.5558 mL	17.7790 mL	35.5581 mL
	5 mM	0.7112 mL	3.5558 mL	7.1116 mL
	10 mM	0.3556 mL	1.7779 mL	3.5558 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.