CC214-1 is a potentially effective mTOR inhibitor that can induce autophagy with IC50 of 0.002 μM. CC214-1 was demonstrated as an in vitro tool compound for exploring mTOR kinase biology. CC214-1 may be used in glioblastoma research.

Physicochemical Properties

Molecular Formula	C20H21N7O2
Molecular Weight	391.43
CAS #	1021920-32-0
Appearance	Typically exists as solid at room temperature
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	mTOR 0.002 μM (IC50)
ln Vitro	Together with rapamycin, CC214-1 (0, 0.1, 1, 2, 5, 10 μM, 8 h; 2 μM, 24 h) inhibits mTORC1 signaling and tumor cell proliferation [1]. Glioblastoma cells are sensitive to growth arrest because of EGFRvIII expression and PTEN loss, which is mediated by CC214-1 (2 μM, 24 h) [1]. In GBM39 cells, CC214-1 (5 μM, 0-48 h) significantly lipidates LC3B-I to LC3B-II isoform and triggers autophagy [1]. CC214-1's IC50 against mTOR is 0.002 μM [2]. T cell activation and the expression of T cell activation markers can be successfully inhibited by CC214-1 (0-10 μM, 4 days) [3].
Cell Assay	Western Blot Analysis[1] Cell Types: glioblastoma Tested Tested Concentrations: 0, 0.1, 1, 2, 5, 10 μM; 2 μM Incubation Duration: 8 h; 24 h Experimental Results: Inhibited mTORC1 signaling in all glioblastoma cell lines tested, potently suppressing rapamycin -resistant 4E-BP1 and mTORC2 signaling. Inhibited mTORC1-dependent 4E-BP1 and S6 phosphorylation in EGFRvIII-expressing glioblastoma cells, as well as blocked glioblastoma cells overexpressing wild-type EGFR. Immunofluorescence[1] Cell Types: glioblastoma Tested Tested Concentrations: 2 μM , 5 μM Incubation Duration: 4 h; 0, 4, 12, 24, 48 h Experimental Results: Induced a transient expression of LC3B-II isoform and the conjugation of Atg12 to Atg5 indicative of the stimulation of the autophagy flux in U87EGFRvIII cell line. Massively lipidated LC3B-I to LC3B-II subtype and induces autophagy in GBM39 cells. Cell Cycle Analysis[3] Cell Types: glioblastoma Tested Tested Concentrations: 0-10 μM Incubation Duration: 4 day Experimental Results: Induced TEE cells in CD4+ and CD8+ T cell subsets .
References	[1]. The mTOR kinase inhibitors, CC214-1 and CC214-2, preferentially block the growth of EGFRvIII-activated glioblastomas. Clin Cancer Res. 2013 Oct 15;19(20):5722-32. [2]. Use of core modification in the discovery of CC214-2, an orally available, selective inhibitor of mTOR kinase. Bioorg Med Chem Lett. 2013 Mar 15;23(6):1588-91. [3]. Effect of mTORC1/mTORC2 inhibition on T cell function: potential role in graft-versus-host disease control. Br J Haematol. 2016 Jun;173(5):754-68.

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	2.5547 mL	12.7737 mL	25.5474 mL
	5 mM	0.5109 mL	2.5547 mL	5.1095 mL
	10 mM	0.2555 mL	1.2774 mL	2.5547 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.