APcK110 is a potent Kit inhibitor that may be utilized in acute myeloid leukemia (AML) research. APcK110 causes apoptosis in AML cells.

Physicochemical Properties

Molecular Formula	C20H16FN3O2
Molecular Weight	349.35834
Exact Mass	349.122
CAS #	1001083-74-4
PubChem CID	24786010
Appearance	Typically exists as solid at room temperature
Density	1.3±0.1 g/cm3
Boiling Point	571.6±45.0 °C at 760 mmHg
Flash Point	299.5±28.7 °C
Vapour Pressure	0.0±1.5 mmHg at 25°C
Index of Refraction	1.636
LogP	3.52
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	4
Heavy Atom Count	26
Complexity	448
Defined Atom Stereocenter Count	0
SMILES	FC1C=CC(=CC=1)C1=C2C=CC(C3C=C(C=C(C=3)OC)OC)=NC2=NN1
InChi Key	CAMGZGMEPKFVMF-UHFFFAOYSA-N
InChi Code	InChI=1S/C20H16FN3O2/c1-25-15-9-13(10-16(11-15)26-2)18-8-7-17-19(23-24-20(17)22-18)12-3-5-14(21)6-4-12/h3-11H,1-2H3,(H,22,23,24)
Chemical Name	6-(3,5-dimethoxyphenyl)-3-(4-fluorophenyl)-2H-pyrazolo[3,4-b]pyridine
Synonyms	APcK110 APcK-110 APcK 110
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	APcK110 (0-500 nM; 0-72 h) suppresses the growth and viability of AML cells. BaF3 cell lines with KIT mutations are preferentially inhibited by APcK110 [1]. In OCI/AML3 cells, APcK110 (500 nM; nightly) causes caspase inhibition. APcK110 (0-500 nM; 30 min) inhibits Kit, Stat3, Stat5, and Akt [1]. Colony-forming cell swelling in AML primary cells is inhibited by APcK110 (50-500 nM; 7 days) [1].
ln Vivo	Mice treated with 500 nM i.p. every other day for 60 days demonstrate anti-AML activity with APcK110 [2].
Cell Assay	Cell proliferation assay[1] Cell Types: OCI/AML3, HMC1.2, BaF3 and BaF3 expressing mutant KIT Cell Tested Concentrations: 0-500 nM Incubation Duration: 0, 24, 48 and 72 hrs (hours) Experimental Results: Inhibition of proliferation at certain doses Dependence method. Western Blot Analysis[1] Cell Types: OCI/AML3 and HMC1.2 Tested Concentrations: 0, 50, 100, 250 and 500 nM Incubation Duration: 30 minutes Experimental Results: Shows dose-dependent inhibition of phosphorylation of Kit, Stat3, Stat5 and Akt , a downstream effector of phosphatidylinositol 3-kinase. Apoptosis analysis[1] Cell Types: OCI/AML3 Cell Tested Concentrations: 500 nM Incubation Duration: Overnight Experimental Results: Induction of apoptosis through activation of the caspase pathway. Cell cycle analysis[1] Cell Types: OCI/AML3 Cell Tested Concentrations: 500 nM Incubation Duration: 2 hrs (hours) Experimental Results: Cells transitioned to sub-G0 after 2 hrs (hours) of incubation.
Animal Protocol	Animal/Disease Models: Eightweeks old female NODSCID (severe combined immunodeficient) mouse, OCI/AML3 xenograft mouse model [2] Doses: 500 nM Route of Administration: intraperitoneal (ip) injection, every other day, for 60 days Experimental Results: Compared with mice injected with PBS , mice treated with the compound lived Dramatically longer.
References	[1]. Kit inhibitor APcK110 induces apoptosis and inhibits proliferation of acute myeloid leukemia cells. Cancer Res. 2009 May 1;69(9):3910-7. [2]. Kit inhibitor APcK110 extends survival in an AML xenograft mouse model. Invest New Drugs. 2011 Oct;29(5):1094-7.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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	2.8624 mL	14.3119 mL	28.6238 mL
	5 mM	0.5725 mL	2.8624 mL	5.7248 mL
	10 mM	0.2862 mL	1.4312 mL	2.8624 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.