CP681301 is a potent CDK5 inhibitor. CP681301 displays antiproliferation activity. CP681301 reduces the expression of CD133, OLIG2, SOX2, KI67, and pCDK5 protein levels in GSCs. CP681301 reduces self-renewal of mouse glioma xenografts. CP681301 displays anti-tumor effects in Drosophila melanogaster.

Physicochemical Properties

Molecular Formula	C17H22N4O
Molecular Weight	298.382783412933
Exact Mass	298.179
CAS #	865317-32-4
PubChem CID	69763070
Appearance	White to light yellow solid powder
LogP	2.6
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	3
Rotatable Bond Count	5
Heavy Atom Count	22
Complexity	375
Defined Atom Stereocenter Count	0
SMILES	C1(CC(NC2C=C(C3CCC3)NN=2)=O)=CC=C(N(C)C)C=C1
InChi Key	PGTNTDKBRSQDNM-UHFFFAOYSA-N
InChi Code	InChI=1S/C17H22N4O/c1-21(2)14-8-6-12(7-9-14)10-17(22)18-16-11-15(19-20-16)13-4-3-5-13/h6-9,11,13H,3-5,10H2,1-2H3,(H2,18,19,20,22)
Chemical Name	N-(5-cyclobutyl-1H-pyrazol-3-yl)-2-[4-(dimethylamino)phenyl]acetamide
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 requires protection from light (avoid light exposure) during transportation and storage.
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	CP6813101 (1 µM; 96 hours) was found to be cytotoxic to tested GSC cultures in different degrees, but not to NHNPs [1]. The expression of CD133, OLIG2, SOX2, KI67, and pCDK5 protein levels in GSCs is decreased by CP6813101 (0, 10, 50 µM; 48 h) [1]. CP681301 (0.5, 1 µM) suppresses the expression of phosphorylation of CREB Ser133 [1].
ln Vivo	CP681301 (1 mM; fed; 10 days) exhibits antitumor activity in adult Drosophila that are 0 to 2 days old [1].
Cell Assay	Cytotoxicity assay[1] Cell Types: NHNP, N12.159, GBM121, GBM39, N08-74 Cell Tested Concentrations: 1 µM Incubation Duration: 96 hrs (hours) Experimental Results: demonstrated varying degrees of cytotoxicity to the tested GSC cultures, but not NHNP is not toxic to (normal) human neural progenitor cells). Western Blot Analysis [1] Cell Types: GBM 121, GBM39 Cell Tested Concentrations: 1 µM Incubation Duration: 48 h Experimental Results: Inhibited the expression of CD133, OLIG2, SOX2 and cell proliferation marker KI67.
Animal Protocol	Animal/Disease Models: 0 to 2 days old adult Drosophila (brat-RNAi tumors) [1] Doses: 1 mM Route of Administration: Fed, 10 days Experimental Results: diminished active phospho-dCdk5 (Y15) in tumor cells, self-renewal capacity Enhanced stem cell properties increased expression of the neuronal marker ElaV in these cells and increased median survival by 2.8-fold.
References	[1]. CDK5 Inhibition Resolves PKA/cAMP-Independent Activation of CREB1 Signaling in Glioma Stem Cells. Cell Rep. 2018 May 8;23(6):1651-1664.

Solubility Data

Solubility (In Vitro)

DMSO : ≥ 100 mg/mL (~335.14 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	3.3514 mL	16.7572 mL	33.5143 mL
	5 mM	0.6703 mL	3.3514 mL	6.7029 mL
	10 mM	0.3351 mL	1.6757 mL	3.3514 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.