MRT00033659 is a potent broad spectrum (a wide range) kinase inhibitor against CK1 (IC50=0.9 µM for CK1δ) and CHK1 (IC50=0.23 µM). MRT00033659 is a pyrazolopyridine analog that induces p53 pathway activation and E2F-1 destabilization.

Physicochemical Properties

Molecular Formula	C15H14N4O
Molecular Weight	266.297862529755
Exact Mass	266.116
CAS #	1401731-54-1
PubChem CID	66555775
Appearance	Off-white to light yellow solid powder
LogP	2
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	3
Rotatable Bond Count	2
Heavy Atom Count	20
Complexity	362
Defined Atom Stereocenter Count	0
SMILES	O=C(C)NC1=CC=CC(=C1)C1C=NC2C(=C(C)NN=2)C=1
InChi Key	YFBPGSDPFRCNOY-UHFFFAOYSA-N
InChi Code	InChI=1S/C15H14N4O/c1-9-14-7-12(8-16-15(14)19-18-9)11-4-3-5-13(6-11)17-10(2)20/h3-8H,1-2H3,(H,17,20)(H,16,18,19)
Chemical Name	N-[3-(3-methyl-2H-pyrazolo[3,4-b]pyridin-5-yl)phenyl]acetamide
Synonyms	MRT-59; MRT 59; MRT59
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 Note: This product is not stable in solution, please use freshly prepared working solution for optimal results.
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	At 5 µM, MRT00033659 (5–40 µM; 48 hours) is enough to drastically lower the number of cells [1]. Massive cell death is induced by MRT00033659 (1-80 µM; 48 hours) beginning at 5 µM [1]. MRT00033659 (0.2-80 µM; 48 hours) destabilizes E2F-1 from 0.2 µM to 5 µM and produces steady and prolonged stabilization of p53, MDM2, and p21 proteins [1]. p38α MAPK is not inhibited by MRT00033659[1].
Cell Assay	Cell Viability Assay[1] Cell Types: A375 Cell Tested Concentrations: 5, 20, 40 µM Incubation Duration: 48 hrs (hours) Experimental Results: Significant reduction in cell number by 5 µM. Apoptosis analysis[1] Cell Types: A375 Cell Tested Concentrations: 1, 5, 10, 20, 40, 80 µM Incubation Duration: 48 hrs (hours) Experimental Results: 5 µM induced massive cell death. Western Blot Analysis [1] Cell Types: A375 Cell Tested Concentrations: 0.2, 1, 5, 10, 20, 40, 80 µM Incubation Duration: 48 hrs (hours) Experimental Results: Induction of p53, MDM2 and p21 proteins stable and stable E2F-1 unstable, From 0.2 µM to 5 µM.
References	[1]. A Casein kinase 1/Checkpoint kinase 1 pyrazolo-pyridine protein kinase inhibitor as novelactivator of the p53 pathway. Bioorg Med Chem Lett. 2013 Oct 15;23(20):5578-85.

Solubility Data

Solubility (In Vitro)

DMSO : ~83.33 mg/mL (~312.92 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 4.17 mg/mL (15.66 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 41.7 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: ≥ 4.17 mg/mL (15.66 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 41.7 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 3: ≥ 4.17 mg/mL (15.66 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 41.7 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	3.7552 mL	18.7758 mL	37.5516 mL
	5 mM	0.7510 mL	3.7552 mL	7.5103 mL
	10 mM	0.3755 mL	1.8776 mL	3.7552 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.