CKG012 is an inhibitor of the Wnt/βcatenin signaling pathway. CGK012 inhibits the release of HMGB1 and the transcription of β-catenin, and is active in ameliorating sepsis induced by cecal ligation and puncture and multiple myeloma cancer.

Physicochemical Properties

Molecular Formula	C20H23NO5
Molecular Weight	357.40
CAS #	2044497-76-7
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

ln Vitro	CGK012 (0-20 μM) inhibits HMGB1 release by inhibiting LPS-induced HMGB1 acetylation and SIRT1 expression, thereby reducing excessive vascular permeability in HUVECs and reducing the expression of pathogen-related molecules such as TLR2/4 without affecting HUVEC cell viability[1]. CGK012 (0-20 μM) improves inflammatory responses by reducing the adhesion and migration of inflammatory immune cells and inhibiting the production of proinflammatory cytokines such as IL-6, TNF-α, β, and transcription factors NF-kB and ERK1/2[1]. CGK012 (0-20 μM) promotes β-catenin phosphorylation and degradation and inhibits the expression of β-catenin-dependent genes, inhibiting the proliferation of multiple myeloma cells RPMI-8226 with an IC50 of 5.08 μM[2].
ln Vivo	CGK012 (0.05-0.53 mg/kg, intravenous injection, double dose) inhibits HMGB1 release and immune cell migration, and improves vascular cell stability and mouse survival in C57BL/6 mice induced by sepsis by cecal ligation and puncture[1].
Cell Assay	Cell Viability Assay[1] Cell Types: HUVEC Concentration: 5-100 μM Incubation Duration: 48 h Experimental Results: Revealed no effects on cell viability. Western Blot Analysis[1] Cell Types: HUVECs, HEK293-FL reporter, RPMI-8226 Concentration: 0-20 μM Incubation Duration: 12 h Experimental Results: Reduced levels of SIRT1 and acetylation of HMGB1, inhibited vascular permeability in HUVECs. Reduced levels of β-catenin in HEK293-FL reporter and RPMI-8226. Cell Migration Assay [1] Cell Types: HUVEC Concentration: 0-20 μM Incubation Duration: 6 h Experimental Results: Inhibited migration of neutrophils through monolayers of HUVECs.
Animal Protocol	Animal/Disease Models:CLP- induced sepsis in C57BL/6 mice[1] Doses: 0.26-0.53 mg/kg Route of Administration: double doses, 12 or 50 h after surgery. Experimental Results: Reduced expression of HMGB1 and improved the survival rate.
References	[1]. Antiseptic Functions of CGK012 against HMGB1-Mediated Septic Responses. Int J Mol Sci. 2024 Mar 4;25(5):2976. [2]. Anti-proliferative activity of CGK012 against multiple myeloma cells via Wnt/β-catenin signaling attenuation. Leuk Res. 2017 Sep;60:103-108.

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	2.7980 mL	13.9899 mL	27.9799 mL
	5 mM	0.5596 mL	2.7980 mL	5.5960 mL
	10 mM	0.2798 mL	1.3990 mL	2.7980 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.