OSU-T315 (ILK-IN-1) is a novel, potent and specific small molecule inhibitor of Integrin-linked kinase (ILK) with an IC50 of 0.6 μM, suppressing cancer cell proliferation in vitro and in vivo and inhibiting PI3K/AKT signaling by dephosphorylation of AKT-Ser473 and other ILK targets (GSK-3β and myosin light chain). OSU-T315 abrogates AKT activation by impeding AKT localization in lipid rafts and triggers caspase-dependent apoptosis in an ILK-independent manner. OSU-T315 causes cell death through apoptosis and autophagy.

Physicochemical Properties

Molecular Formula	C₃₀H₃₀F₃N₅O
Molecular Weight	533.59
Exact Mass	533.24
CAS #	2070015-22-2
PubChem CID	118986645
Appearance	White to off-white solid powder
Density	1.3±0.1 g/cm3
Boiling Point	739.2±60.0 °C at 760 mmHg
Flash Point	400.9±32.9 °C
Vapour Pressure	0.0±2.4 mmHg at 25°C
Index of Refraction	1.615
LogP	4.65
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	7
Heavy Atom Count	39
Complexity	767
Defined Atom Stereocenter Count	0
InChi Key	AJLOJUFSIDSBNN-UHFFFAOYSA-N
InChi Code	InChI=1S/C30H30F3N5O/c1-34-29(39)15-14-27-20-28(36-38(27)26-12-10-25(11-13-26)37-18-16-35-17-19-37)23-4-2-21(3-5-23)22-6-8-24(9-7-22)30(31,32)33/h2-13,20,35H,14-19H2,1H3,(H,34,39)
Chemical Name	N-methyl-3-[2-(4-piperazin-1-ylphenyl)-5-[4-[4-(trifluoromethyl)phenyl]phenyl]pyrazol-3-yl]propanamide
Synonyms	OSUT315 OSU T315 OSU-T315
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	Compound 22, OSU-T315 (IC50 ranging from 1-2.5 μM), demonstrates excellent potency in vitro against a panel of prostate and breast cancer cell lines [1]. The expression of YB-1, HER2, and EGFR can be decreased by OSU-T315 (0-2.5 μM; 24 hours); it also exhibits a dose-dependent influence on phosphorylated p38 and ERK1/2 and a modest inhibitory effect on phosphorylated S6 levels. sexual inhibition, whereas PC-3 cells show no change in phosphorylated JNK [1]. ILK inhibition is used by OSU-T315 (0–4 μM; 24 hours) to promote autophagy [1].
ln Vivo	PC-3 xenograft tumor growth is inhibited by OSU-T315 (oral gavage; 25 mg/kg, 50 mg/kg; once daily; 35 days) [1]. ?In mice, no additional noteworthy harm was noted [1].
Cell Assay	Western Blot Analysis[1] Cell Types: PC-3 cells; MDA-MB-231 cells Tested Concentrations: 1μM, 2μM, 3μM, 4μM; 0.5μM, 1μM, 1.5μM, 2μM, 2.5μM Incubation Duration: 24 hrs (hours) Experimental Results: Exhibits a dose-dependent reduction in the phosphorylation of pS6, ERK and p38 in PC-3 cells and MDA-MB-231 cells. Cell viability assay[1] Cell Types: Prostate cancer cells: LNCaP, PC-3; Breast cancer cells: MDA-MB-231, MDA-MB-468, SKBR3, MCF-7; PrEC and MEC cell Tested Concentrations: 0-5 μM Incubation Duration: 24 hrs (hours) Experimental Results: Inhibition of cancer cell viability in breast and prostate cancer cells (IC (50), 1-2.5 μM). Apoptosis analysis[1] Cell Types: PC-3 Cell Tested Concentrations: 1 μM, 2 μM, 3 μM, 4 μM Incubation Duration: 24 hrs (hours) Experimental Results: Induction of accumulation of LC3-II and PARP cleavage.
Animal Protocol	Animal/Disease Models: Male NCr athymic nude mice with PC-3 tumor xenografts Doses: 25 mg/kg; 50 mg/kg Route of Administration: po (oral gavage); daily single; 35 days Experimental Results: After 35 days of treatment, relative Tumor growth was inhibited compared to vehicle control (48% and 62% inhibition at 25 mg/kg and 50 mg/kg, respectively).
References	[1]. Identification and Characterization of a Novel Integrin-Linked Kinase Inhibitor.J Med Chem. 2011 Sep 22; 54(18): 6364–6374. [2]. OSU-T315: a novel targeted therapeutic that antagonizes AKT membrane localization and activation of chronic lymphocytic leukemia cells. Blood. 2015 Jan 8;125(2):284-95.

Solubility Data

Solubility (In Vitro)

DMSO : ≥ 260 mg/mL (~487.27 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.17 mg/mL (4.07 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 21.7 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. Solubility in Formulation 2: ≥ 2.17 mg/mL (4.07 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 21.7 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	1.8741 mL	9.3705 mL	18.7410 mL
	5 mM	0.3748 mL	1.8741 mL	3.7482 mL
	10 mM	0.1874 mL	0.9370 mL	1.8741 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.