CEP-28122 mesylate salt is a diaminopyrimidine analogue that is a specific and orally bioactive ALK inhibitor (antagonist) with IC50 of 1.9 nM for recombinant ALK kinase activity. CEP-28122 displays anticancer effect in ALK-positive experimental models of human cancer. CEP-28122 has good pharmacodynamic and pharmacokinetic activities.

Physicochemical Properties

Molecular Formula	C29H39CLN6O6S
Molecular Weight	635.17
CAS #	2070009-30-0
Related CAS #	CEP-28122;1022958-60-6
PubChem CID	91885507
Appearance	White to off-white solid powder
Hydrogen Bond Donor Count	4
Hydrogen Bond Acceptor Count	11
Rotatable Bond Count	7
Heavy Atom Count	43
Complexity	951
Defined Atom Stereocenter Count	5
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: Please store this product in a sealed and protected environment, avoid exposure to moisture.
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	Treatment with CEP-28122 mesylate salt (3–3000 nM; 48 hours) inhibits the development of Karpas-299 and Sup-M2 cells in culture in a concentration-dependent manner and is associated with concentration-related caspase 3/7 activation[1]. Phosphorylation of potential ALK downstream effectors is significantly suppressed in Sup-M2 cells after treatment with CEP-28122 mesylate salt (30–1000 nM; 2 hours), suggesting that individual ALK fusion proteins mediate the downstream signaling pathways[1].
ln Vivo	In Sup-M2 subcutaneous tumor xenografts in SCID mice, CEP-28122 mesylate salt (3–30 mg/kg; oral gavage; twice daily; 12 days) exhibits dose-dependent antitumor activity. In contrast, CEP-28122 has no antitumor activity in nu/nu mice bearing HCT116, indicating that the antitumor activity of CEP-28122 in NPM-ALK–positive Sup-M2 tumor models is the result of persistent NPM-ALK inhibition in tumors [1].
Cell Assay	Cell Cytotoxicity Assay[1] Cell Types: Karpas-299, Sup-M2, Toledo and HuT-102 cells Tested Concentrations: 10 nM, 100 nM, 1000 nM, 10000 nM Incubation Duration: 48 hrs (hours) Experimental Results: Treatment led to concentration-dependent growth inhibition of Karpas-299 and Sup-M2 cells in culture. Western Blot Analysis [1] Cell Types: Sup-M2 cells Tested Concentrations: 30 nM, 100 nM, 300 nM, 1000 nM Incubation Duration: 2 hrs (hours) Experimental Results: Resulted in substantial suppression of phosphorylation of putative downstream effectors of ALK, including Stat-3, Akt, and ERK1/2 in Sup-M2 cells.
Animal Protocol	Animal/Disease Models: Female SCID (severe combined immunodeficient) mouse bearing Sup -M2 subcutaneous (sc) tumor xenografts and nu/nu (nude) mice bearing HCT116 aged 6-8 week old[1] Doses: 3 mg/kg, 10 mg/kg and 30 mg/kg Route of Administration: po (oral gavage); twice a day; 12 days Experimental Results: CEP-28122 produced dose-dependent antitumor activity in Sup-M2 subcutaneous (sc) tumor xenografts in SCID (severe combined immunodeficient) mouse. In contrast, CEP-28122 had no antitumor activity in nu/nu (nude) mice bearing HCT116.
References	[1]. CEP-28122, a highly potent and selective orally active inhibitor of anaplastic lymphoma kinase with antitumor activity in experimental models of human cancers. Mol Cancer Ther. 2012 Mar;11(3):670-9.

Solubility Data

Solubility (In Vitro)

DMSO :~6.4 mg/mL (~10.08 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	1.5744 mL	7.8719 mL	15.7438 mL
	5 mM	0.3149 mL	1.5744 mL	3.1488 mL
	10 mM	0.1574 mL	0.7872 mL	1.5744 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.