T56-LIMKi (also known as T5601640) is a potent and selective inhibitor of LIMK2 (LIM kinase 2) with an IC50 of 35.2 μM in a cell (Panc-1 cells) assay. T56-LIMKi exhibits little to no cross-reactivity with LIMK1 and inhibits LIMK2 with high specificity. T56-LIMKi inhibits the growth of multiple cancerous cell lines, such as those of glioma, schwannoma, and pancreatic cancer, by lowering the levels of phosphorylated cofilin (p-cofilin). T56-LIMKi lowered p-cofilin levels and tumor size in a panc-1 xenograft model in naked mice. LIM kinases are significant regulators of the cell cytoskeleton and are involved in the development, metastasis, and expression of cancer as well as disorders of the nervous system. T56-LIMKi may therefore be developed as a candidate medication for neuronal disorders and cancer therapy.

Physicochemical Properties

Molecular Formula	C19H14F3N3O3
Molecular Weight	389.33
Exact Mass	389.098
Elemental Analysis	C, 58.62; H, 3.62; F, 14.64; N, 10.79; O, 12.33
CAS #	924473-59-6
Related CAS #	924473-59-6
PubChem CID	9438169
Appearance	White to off-white solid powder
Density	1.4±0.1 g/cm3
Boiling Point	403.4±45.0 °C at 760 mmHg
Flash Point	197.8±28.7 °C
Vapour Pressure	0.0±0.9 mmHg at 25°C
Index of Refraction	1.618
LogP	2.85
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	4
Heavy Atom Count	28
Complexity	572
Defined Atom Stereocenter Count	0
SMILES	O=C(C1=CC(C)=NO1)NC1C=C(C(NC2C=C(C(F)(F)F)C=CC=2)=O)C=CC=1
InChi Key	XVOKFRPKSAWELK-UHFFFAOYSA-N
InChi Code	InChI=1S/C19H14F3N3O3/c1-11-8-16(28-25-11)18(27)24-14-6-2-4-12(9-14)17(26)23-15-7-3-5-13(10-15)19(20,21)22/h2-10H,1H3,(H,23,26)(H,24,27)
Chemical Name	3-methyl-N-[3-[[3-(trifluoromethyl)phenyl]carbamoyl]phenyl]-1,2-oxazole-5-carboxamide
Synonyms	T56LIMKi; T56 LIMKi; T56-LIMKi; T-5601640; T 5601640; T5601640
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

Targets	LIMK2
ln Vitro	T56-LIMKi efficiently inhibits the growth of ST88-14, U87, Panc-1 cells, A549 lung cancer cells with IC50 values of 18.3, 7.4, 35.2 and 90 μM, respectively. T56-LIMKi inhibits the growth of multiple cancerous cell lines, such as those of glioma, schwannoma, and pancreatic cancer, by lowering the levels of phosphorylated cofilin (p-cofilin)[1]. It inhibits the growth, migration, and anchorage-independent colony formation of tumor cells in soft agar as well as actin severance caused by the phosphorylation of cofilin. In NF1−/−MEFs, T56-LIMKi (10-50 μM) has an IC50 of 30 μM and decreases p-cofilin in a dose-dependent manner. Interestingly, the inhibitor has no effect on total cofil concentrations. 50 T There is a statistically significant decrease in the number of cells displaying stress fibers when T56-LIMKi is used[2].
ln Vivo	T56-LIMKi can cause a decrease in Panc-1 tumor size and an inhibition of cofilin phosphorylation in vivo. Tumor volume in mice given T56-LIMKi (60 mg/kg) is significantly lower than in control mice[1].
Enzyme Assay	T56-LIMKi (also known as T5601640) is a powerful and focused inhibitor of LIMK2 (LIM kinase 2), exhibiting an IC50 of 35.2 μM in a cell assay using Panc-1 cells.
Cell Assay	T56-LIMKi can cause Panc-1 tumor shrinkage and cofilin phosphorylation inhibition in vivo. Tumor volume in mice treated with T56-LIMKi (60 mg/kg) is significantly lower than in control mice[1].
Animal Protocol	Mice: In a 0.5% carboxymethylcellulose solution, T56-LIMKi is dissolved. Panc-1 xenografts are injected into mice. Seven days later, treatment is initiated. Mice in the control group are given only the vehicle (0.5% CMC) in the gavage, while mice in the two experimental groups are given daily oral non-toxic doses of T56-LIMKi (30 or 60 mg/kg)[1].
References	[1]. Novel LIMK2 Inhibitor Blocks Panc-1 Tumor Growth in a mouse xenograft model. Oncoscience. 2014 Jan 1;1(1):39-48. eCollection 2014. [2]. Computer-based identification of a novel LIMK1/2 inhibitor that synergizes with salirasib to destabilize the actin cytoskeleton. Oncotarget. 2012 Jun;3(6):629-39.

Solubility Data

Solubility (In Vitro)

DMSO:≥ 40 mg/mL Water: Ethanol:

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.42 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	2.5685 mL	12.8426 mL	25.6852 mL
	5 mM	0.5137 mL	2.5685 mL	5.1370 mL
	10 mM	0.2569 mL	1.2843 mL	2.5685 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.