T025 is an orally bioactive Cdc2-like kinase (CLK) inhibitor (antagonist) with Kds of 4.8, 0.096, 6.5, 0.61, 0.074, 1.5 and 32 nM for CLK1, CLK2, CLK3, CLK4, DYRK1A, DYRK1B and DYRK2 respectively. T025 induces caspase-3/7-mediated apoptosis. T025 reduces CLK-dependent phosphorylation. T025 has antiproliferation activity in both blood and solid cancer cell lines (IC50= 30-300 nM). T025 has anti-tumor efficacy and is mainly used in MYC-driven disease study.

Physicochemical Properties

Molecular Formula	C21H18N8
Molecular Weight	382.42122220993
Exact Mass	382.165
CAS #	2407433-00-3
PubChem CID	133082038
Appearance	Yellow to green solid powder
LogP	3
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	5
Heavy Atom Count	29
Complexity	527
Defined Atom Stereocenter Count	0
SMILES	N1C=C(C2C=CC3C(=CC=CN=3)C=2)C2=C(N=C(NC)N=C12)NCC1N=CC=CN=1
InChi Key	PMWVYONICYRLNY-UHFFFAOYSA-N
InChi Code	InChI=1S/C21H18N8/c1-22-21-28-19-18(20(29-21)27-12-17-24-8-3-9-25-17)15(11-26-19)13-5-6-16-14(10-13)4-2-7-23-16/h2-11H,12H2,1H3,(H3,22,26,27,28,29)
Chemical Name	2-N-methyl-4-N-(pyrimidin-2-ylmethyl)-5-quinolin-6-yl-7H-pyrrolo[2,3-d]pyrimidine-2,4-diamine
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	CLK2 0.096 nM (Kd) CLK4 0.61 nM (Kd) CLK1 4.8 nM (Kd) CLK3 6.5 nM (Kd) DYRK1A 0.074 nM (Kd) DYRK1B 1.5 nM (Kd) DYRK2 32 nM (Kd)
ln Vitro	In a dose-dependent manner, T025 (0-1000 nM; 72 hours) strongly reduces the proliferation of MDA-MB-468 cells[1]. Phosphorylation levels in MDA-MB-468 cells are decreased by T025 (0-1000 nM; 6 hours)[1].
ln Vivo	T025 (50 mg/kg; po; 2, 4, 8 hours) suppresses the phosphorylation of CLK-dependent genes and causes exon skipping in a variety of genes in Balb/c nude mice (7 to 8 week old females)[1]. T025 (50 mg/kg; po; twice daily on two days per week, for three weeks) reduces the formation of tumors in MDA-MB-468 xenograft mice without affecting body weight in Balb/c nude mice (seven to eight week old females) [1].
Cell Assay	Cell Viability Assay[1] Cell Types: MDA-MB-468 cells Tested Concentrations: 1, 10, 100 and 1000 nM Incubation Duration: 72 hrs (hours) Experimental Results: Resulted in concentration dependent growth inhibition. Western Blot Analysis[1] Cell Types: MDA-MB-468 cells Tested Concentrations: 0, 10, 30, 100, 300 and 1000 nM Incubation Duration: 6 hrs (hours) Experimental Results: diminished both pCLK2 and CLK2.
Animal Protocol	Animal/Disease Models: Balb/c nude mice (7 to 8 week-old females)[1] Doses: 50 mg/kg Route of Administration: Oral administration; 2, 4, 8 hrs (hours). Experimental Results: pCLK2 detected with immunohistochemistry and immunoblotting diminished, by a reduction in the RPS6KB1 exon 7 and BCLAF1 exon 11 percentage splice-in (PSI) values. Animal/Disease Models: Balb/c nude mice ( 7 to 8 week-old females)[1] Doses: 50 mg/kg Route of Administration: Oral administration; twice (two times) daily on 2 days per week, for 3 weeks. Experimental Results: Suppressed tumor growth and < 10% nadir body weight loss.
References	[1]. Anti-tumor efficacy of a novel CLK inhibitor via targeting RNA splicing and MYC-dependent vulnerability. EMBO Mol Med. 2018 Jun;10(6):e8289.

Solubility Data

Solubility (In Vitro)	DMSO : 8.25 mg/mL (21.57 mM)
Solubility (In Vivo)	Solubility in Formulation 1: 10 mg/mL (26.15 mM) in 0.5% CMC-Na/saline water (add these co-solvents sequentially from left to right, and one by one), suspension solution. 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	2.6149 mL	13.0746 mL	26.1493 mL
	5 mM	0.5230 mL	2.6149 mL	5.2299 mL
	10 mM	0.2615 mL	1.3075 mL	2.6149 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.