PCC0208017 is a potent inhibitor of microtubule affinity-regulated kinase MARK3/MARK4 with IC50s of 1.8 and 2.01 nM respectively. PCC0208017 has much lower inhibitory effect against MARK1 and MARK2, with IC50s of 31.4 and 33.7 nM, respectively. PCC0208017 inhibits glioma progression in vitro and in vivo. PCC0208017 disrupts microtubule dynamics and induces G2/M phase cell cycle arrest and apoptosis. PCC0208017 has potent anticancer effect and shows good BBB permeability.

Physicochemical Properties

Molecular Formula	C19H20F3N7
Molecular Weight	403.40
Exact Mass	403.173
CAS #	2623158-64-3
PubChem CID	163321087
Appearance	Off-white to light yellow solid powder
LogP	3
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	5
Heavy Atom Count	29
Complexity	534
Defined Atom Stereocenter Count	0
SMILES	C1(C(NCC2C=CNN=2)=NC(NC2=CC=C3CN(C)CCC3=C2)=NC=1)C(F)(F)F
InChi Key	PIESSZHWYCPHQD-UHFFFAOYSA-N
InChi Code	InChI=1S/C19H20F3N7/c1-29-7-5-12-8-14(3-2-13(12)11-29)26-18-24-10-16(19(20,21)22)17(27-18)23-9-15-4-6-25-28-15/h2-4,6,8,10H,5,7,9,11H2,1H3,(H,25,28)(H2,23,24,26,27)
Chemical Name	2-N-(2-methyl-3,4-dihydro-1H-isoquinolin-6-yl)-4-N-(1H-pyrazol-5-ylmethyl)-5-(trifluoromethyl)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

ln Vitro	In addition to lowering Tau phosphorylation, PCC0208017 suppresses MARK3 and MARK4 activity[1]. Tau phosphorylation is reduced upon treatment with PCC0208017 (1–5 μM; 24 hours)[1]. Glioma cells are inhibited from proliferating by PCC0208017 (3–21 μM; 24 hours)[1].
ln Vivo	In vivo, PCC0208017 exhibits strong anticancer activity and good blood-brain barrier permeability. In a dose-dependent manner, PCC0208017 (50 and 100 mg/kg) suppresses the formation of xenograft tumors generated from GL261 cells. The corresponding inhibition rates are 56.15% and 70.32%. Temozolomide (TMZ; 100 mg/kg)'s anti-tumor efficacy is markedly increased when PCC0208017 is co-treated at a dosage of 50 mg/kg. Tumor inhibition rates rise from 34.15% (TMZ alone) to 83.5% (TMZ+PCC0208017)[1]. A single oral dosage of 50 mg/kg was sufficient to detect PCC0208017 in both the brain and plasma after that. Tmax in plasma is 0.833 h, while Cmax is 1.36 μg/mL. The brain has a Tmax of 0.833 hours and a Cmax of 0.14 μg/mL[1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: The glioma cell lines GL261, U87-MG, U251 Tested Concentrations: 0, 3, 6, 9, 12, 15, 18, 21 μM Incubation Duration: 24 hrs (hours) Experimental Results: The IC50 values for GL261, U87-MG and U251 were calculated as 2.77, 4.02 and 4.45 μM, respectively. Cell Proliferation Assay[1] Cell Types: Tested Concentrations: Glioma cell lines GL261 and U251 1, 2, 5 μM Incubation Duration: 24 hrs (hours) Experimental Results: diminished the phosphorylation of Tau.
Animal Protocol	Animal/Disease Models: C57BL/6 mice bearing murine glioma GL261 xenograft tumor[1] Doses: 50 mg/kg and 100 mg/kg (suspended in a 0.5% methylcellulose solution)[1]. Route of Administration: Orally administered every day at a volume of 10 mL/kg Experimental Results: Inhibited GL261 cells growth in xenograft mouse model.
References	[1]. PCC0208017, a novel small-molecule inhibitor of MARK3/MARK4, suppresses glioma progression in vitro and in vivo. Acta Pharm Sin B.2020 Feb;10(2):289-300.

Solubility Data

Solubility (In Vitro)

DMSO : 125 mg/mL (309.87 mM)

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.4789 mL	12.3946 mL	24.7893 mL
	5 mM	0.4958 mL	2.4789 mL	4.9579 mL
	10 mM	0.2479 mL	1.2395 mL	2.4789 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.