CX-6258 is a novel, potent, selective and orally bioactive pan-Pim kinase inhibitor with IC50 of 5 nM, 25 nM and 16 nM for Pim1, Pim2, and Pim3, respectively. At CX-6258 showed robust antiproliferative potencies against all cell lines tested derived from human solid tumors and hematological malignancies. In mechanistic cellular assays with MV-4-11 human AML cells, caused dose-dependent inhibition of the phosphorylation of 2 pro-survival proteins, Bad and 4E-BP1, at the Pim kinase specific sites S112 and S65 and T37/46, respectively.

Physicochemical Properties

Molecular Formula	C26H24CLN3O3
Molecular Weight	461.94
Exact Mass	461.15
CAS #	1202916-90-2
Related CAS #	CX-6258 hydrochloride hydrate;1353858-99-7
PubChem CID	44545852
Appearance	Yellow to orange solid powder
Density	1.3±0.1 g/cm3
Boiling Point	697.8±55.0 °C at 760 mmHg
Flash Point	375.8±31.5 °C
Vapour Pressure	0.0±2.2 mmHg at 25°C
Index of Refraction	1.646
LogP	4.33
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	3
Heavy Atom Count	33
Complexity	774
Defined Atom Stereocenter Count	0
SMILES	CN1CCCN(CC1)C(=O)C2=CC=CC(=C2)C3=CC=C(O3)/C=C/4\C5=C(C=CC(=C5)Cl)NC4=O
InChi Key	KGBPLKOPSFDBOX-CJLVFECKSA-N
InChi Code	InChI=1S/C26H24ClN3O3/c1-29-10-3-11-30(13-12-29)26(32)18-5-2-4-17(14-18)24-9-7-20(33-24)16-22-21-15-19(27)6-8-23(21)28-25(22)31/h2,4-9,14-16H,3,10-13H2,1H3,(H,28,31)/b22-16+
Chemical Name	(3E)-5-chloro-3-[[5-[3-(4-methyl-1,4-diazepane-1-carbonyl)phenyl]furan-2-yl]methylidene]-1H-indol-2-one
Synonyms	CX6258; CX 6258;CX-6258
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	Two pro-survival proteins, Bad and 4E-BP1, are phosphorylated at Pim kinase specific sites S112, S65, and T37/46, respectively, and are inhibited by CX-6258 in a dose-dependent manner[1]. In PC3 cells, CX-6258 treatment (12 mM, 3 h) reduces steady-state levels of ectopic NKX3.1[2]. The half-life of NKX3.1 is significantly shortened by CX-6258 treatment[2].
ln Vivo	In two tumor models driven by Pim kinases, CX-6258 (50–100 mg/kg; po; daily; for 21 days) shows strong in vivo efficacy[1].
Cell Assay	Western Blot Analysis[1] Cell Types: MV-4-11 human AML cells Tested Concentrations: 0.1 μM, 1 μM, 10 μM Incubation Duration: 2 hrs (hours) Experimental Results: Caused dose dependent inhibition of the phosphorylation of two pro-survival proteins, Bad and 4E-BP1, at the Pim kinase specific sites S112 and S65 and T37/46, respectively.
Animal Protocol	Animal/Disease Models: Nude mice, MV-4-11 xenograft Doses: 50 mg/kg, 100 mg/kg Route of Administration: Oral administration; one time/day; over a period of 21 days Experimental Results: demonstrated dose dependent efficacy, with a 50 mg/kg dose producing 45% tumor growth inhibition ( TGI) and a 100 mg/kg dose producing 75% TGI.
References	[1]. Mustapha Haddach, Jerome Michaux, Michael K, Discovery of CX-6258. A Potent, Selective, and Orally Efficacious pan-Pim Kinases Inhibitor. ACS Med. Chem. Lett., 2012, 3 (2), pp 135-139. [2]. Padmanabhan A, Gosc EB, Bieberich CJ. Stabilization of the prostate-specific tumor suppressor NKX3.1 by the oncogenic protein kinase Pim-1 in prostate cancer cells. J Cell Biochem. 2013 May;114(5):1050-7.

Solubility Data

Solubility (In Vitro)

DMSO: 10 mM Water:<1 mg/mL Ethanol:<1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: 20 mg/mL (43.30 mM) in 15% Cremophor EL + 85% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication (<60°C). Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.75 mg/mL (5.95 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 27.5 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.1648 mL	10.8239 mL	21.6478 mL
	5 mM	0.4330 mL	2.1648 mL	4.3296 mL
	10 mM	0.2165 mL	1.0824 mL	2.1648 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.