CDK8-IN-11 is a potent and specific inhibitor of CDK8 with IC50 of 46 nM. CDK8-IN-11 inhibits the WNT/β-catenin signaling pathway. CDK8-IN-11 may be used in colon cancer research.

Physicochemical Properties

Molecular Formula	C19H15F3N4O2
Molecular Weight	388.343214273453
Exact Mass	388.114
CAS #	2839338-28-0
Related CAS #	CDK8-IN-11 hydrochloride
PubChem CID	165412810
Appearance	Light yellow to yellow solid powder
LogP	3.5
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	4
Heavy Atom Count	28
Complexity	512
Defined Atom Stereocenter Count	0
InChi Key	MBIXOXMOHXFUFZ-UHFFFAOYSA-N
InChi Code	InChI=1S/C19H15F3N4O2/c20-19(21,22)12-2-1-3-14(10-12)26-18(27)25-13-4-6-15(7-5-13)28-16-8-9-17(23)24-11-16/h1-11H,(H2,23,24)(H2,25,26,27)
Chemical Name	1-[4-(6-aminopyridin-3-yl)oxyphenyl]-3-[3-(trifluoromethyl)phenyl]urea
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	CDK8 46 nM (IC50)
ln Vitro	Compound 29, 200 nM of CDK8-IN-11 exhibits 73.6% inhibitory actions against CDK8 [1]. Cell proliferation in HCT-116, HHT-29, SW480, CT-26, and GES-1 cells is inhibited by CDK8-IN-11 (0-50 μM, 48 h)[1]. In HCT-116 cells, CDK8-IN-11 (0–4 μM, 48 h) suppresses the phosphorylation of STAT1 at Ser727, which is mediated by CDK8[1]. In HCT-116 cells, CDK8-IN -11 (0–4 μM, 24 h) deregulates β-catenin-mediated transcription and reduces canonical WNT/β-catenin signaling pathways[1]. In HCT-116 cells, CDK8-IN-11 (0.5-2 μM, 48 h) increases the proportion of G1 phase cells[1]. HCT-116 cells' resistance to sorafenib (HY-10201) is reversed by CDK8-IN-11 (0–4 μM)[1].
ln Vivo	In CT-26 xenograft mice, CDK8-IN-11 (compound 29, 10 and 40 mg/kg, po) inhibits the growth of tumors[1]. After seven days, CDK8-IN-11 mice (1000 mg/kg, oral gavage, ICR mice) exhibit no observable aberrant behavior[1]. Rats treated with 10 mg/kg po or 2 mg/kg iv of CDK8-IN-11 exhibit moderate permeability, as measured by an apparent permeability coefficient of 1.8 × 10−6 cm/s [1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: HCT-116, HHT-29, SW480, CT-26, GES-1 cells Tested Concentrations: 0.08, 0.4, 2, 10, and 50 μM Incubation Duration: 48 h Experimental Results: Inhibited cell proliferation with IC50 values of 1.2, 0.7, 2.4, 5.5, 62.7 nM respectively. Western Blot Analysis[1] Cell Types: HCT-116 cell Tested Concentrations: 0, 1, 2, 4 μM Incubation Duration: 48 h Experimental Results: Inhibited the phosphorylation of STAT1 at Ser727 without affecting the JAK-regulated phosphorylation at Tyr701. Cell Cycle Analysis[1] Cell Types: HCT-116 cell Tested Concentrations: 0.5-2 μM Incubation Duration: 48 h Experimental Results: Increased the number of cells in the G1 phase with an obvious diminished percentage of cells in the G2/M and S phase in HCT-116 cells.
Animal Protocol	Animal/Disease Models: CT-26 xenograft mice[1] Doses: 10 and 40 mg/kg Route of Administration: Oral adminstration (po) Experimental Results: diminished the tumor volume, diminished β-catenin and c-Myc level in tumor. Animal/Disease Models: Rats (pharmacokinetic/PK assay)[1] Doses: 10 mg/kg (po), 2 mg/kg (iv) Route of Administration: Oral adminstration (po ) or intravenous (iv)injection Experimental Results: pharmacokinetic/PK profile of CDK8-IN-11 (compound 29). dose (mg/kg) T1/2 (h) Tmax (h) Cmax (ng/mL) F (%) 10 ( po) 1.1 0.8 453 31.7 2 (iv) 0.5 318
References	[1]. Design and Synthesis of a 2-Amino-pyridine Derivative as a Potent CDK8 Inhibitor for Anti-colorectal Cancer Therapy. J Med Chem. 2022 Sep 20.

Solubility Data

Solubility (In Vitro)

DMSO : 250 mg/mL (643.77 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	2.5751 mL	12.8753 mL	25.7506 mL
	5 mM	0.5150 mL	2.5751 mL	5.1501 mL
	10 mM	0.2575 mL	1.2875 mL	2.5751 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.