TC-G 24 (Compound 24) is a potent and specific inhibitor of glycogen synthase kinase 3β (GSK-3β) with IC50 of 17.1 nM. TC-G 24 is able to cross the BBB (blood-brain barrier) and may be utilized to study many diseases, like type 2 diabetes, stroke, Alzheimer's disease (AD) and other related diseases.

Physicochemical Properties

Molecular Formula	C15H11CLN4O3
Molecular Weight	330.73
Exact Mass	330.052
CAS #	1257256-44-2
PubChem CID	49857866
Appearance	Typically exists as solid at room temperature
LogP	4.295
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	3
Heavy Atom Count	23
Complexity	414
Defined Atom Stereocenter Count	0
SMILES	CC1=C(C=C(C=C1)NC2=NN=C(O2)C3=CC=C(C=C3)[N+](=O)[O-])Cl
InChi Key	JYCNWQGNEJYDQS-UHFFFAOYSA-N
InChi Code	InChI=1S/C15H11ClN4O3/c1-9-2-5-11(8-13(9)16)17-15-19-18-14(23-15)10-3-6-12(7-4-10)20(21)22/h2-8H,1H3,(H,17,19)
Chemical Name	N-(3-chloro-4-methylphenyl)-5-(4-nitrophenyl)-1,3,4-oxadiazol-2-amine
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	TC-G 24 (compound 24) attaches itself to GSK-3β's ATP binding site [1]. Human embryonic kidney (HEK) 293T cells are resistant to TPP1 degradation caused by FBW7α when exposed to TC-G 24 (1 µM, 4 hours) [2].
ln Vivo	Compound 24, also known as TC-G 24, can penetrate the blood-brain barrier and dramatically raise the amount of liver glycogen in a dose-dependent manner when administered intraperitoneally once at a dose of 0–15 mg/kg [1].
Cell Assay	Western blot analysis[2] Cell Types: 293T Cell Tested Concentrations: 1 μM Incubation Duration: 4 hrs (hours) Experimental Results: Blocks FBW7α-mediated TPP1 degradation
Animal Protocol	Animal/Disease Models: Sixweeks old male C57BL/6N mice, average weight 22 g[1] Doses: 1, 5 and 15 mg/kg Route of Administration: intraperitoneal (ip) injection, one time Experimental Results:Liver glycogen content increased Dramatically, showing There was no significant toxicity in a dose-dependent manner. At all three doses tested, higher concentrations were detected in the brain than in plasma (38 ± 6, 113 ± 54, and 286 ± 58 ng/g brain tissue at 1, 5, and 15 mg/kg concentrations, respectively ).
References	[1]. Discovery of novel GSK-3β inhibitors with potent in vitro and in vivo activities and excellent brain permeability using combined ligand- and structure-based virtual screening. J Med Chem. 2010 Dec 23;53(24):8534-45. [2]. FBW7 Mediates Senescence and Pulmonary Fibrosis through Telomere Uncapping. Cell Metab. 2020 Nov 3;32(5):860-877.e9.

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	3.0236 mL	15.1181 mL	30.2361 mL
	5 mM	0.6047 mL	3.0236 mL	6.0472 mL
	10 mM	0.3024 mL	1.5118 mL	3.0236 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.