M50054 is a potent apoptosis inhibitor. M50054 inhibits Etoposide-induced caspase-3 activation in U937 cells with IC50 of 79 μg/mL. M50054 does not directly inhibit caspase-3 enzyme activity. M50054 may be utilized to study hepatitis caused by anti-Fas antibodies and chemotherapy-induced alopecia.

Physicochemical Properties

Molecular Formula	C13H16O4
Molecular Weight	236.26
Exact Mass	236.105
CAS #	54135-60-3
PubChem CID	2748618
Appearance	Off-white to light yellow solid powder
Density	1.227 g/cm3
Boiling Point	461.2ºC at 760 mmHg
Flash Point	199ºC
Index of Refraction	1.52
LogP	1.253
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	2
Heavy Atom Count	17
Complexity	315
Defined Atom Stereocenter Count	0
SMILES	C1CC(=O)C(CC2C(=O)CCCC2=O)C(=O)C1
InChi Key	ILUMEPMGPCKGHH-UHFFFAOYSA-N
InChi Code	InChI=1S/C13H16O4/c14-10-3-1-4-11(15)8(10)7-9-12(16)5-2-6-13(9)17/h8-9H,1-7H2
Chemical Name	2-[(2,6-dioxocyclohexyl)methyl]cyclohexane-1,3-dione
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	M50054 suppresses the dose-dependent activation of caspase-3 by etoposide. The inhibitory concentration of M50054 against the activation of caspase-3 produced by etoposide in U937 cells is 79 μg/mL. M50054, however, does not directly block caspase-3's enzymatic activity at any dose up to 1000 μg/mL[1]. With an IC50 of 67 μg/mL, M50054 (1-300 μg/mL) suppresses the dose-dependent induction of cell death of human Fas-expressing WC8 cells by the soluble human Fas ligand[1]. The mortality of U937 cells caused by etoposide (10 μg/mL) is inhibited by M50054 (30-300 μg/mL) in a dose-dependent manner; M50054's IC50 value against Etoposide-induced U937 cell death is 130 μg/mL. M50054's IC50 value against etoposide-induced U937 cell DNA fragmentation was 54 μg/mL[1].
ln Vivo	The increase of aspartate and alanine aminotransferases is dose-dependently inhibited by M50054 (10–300 mg/kg; oral given 30 min before anti-Fas antibody injection)[1]. The topical medication M50054, which is applied once daily from the fifth day after delivery until the fourteenth day, considerably reduces the symptoms of alopecia (hair loss)[1].
Cell Assay	Cell Proliferation Assay[1] Cell Types: Human Fas-expressing WC8 cells Tested Concentrations: 1, 3, 10, 30, 100, 300 μg/mL Incubation Duration: Pre-incubated for 30 min Experimental Results: Inhibited soluble human Fas ligand-induced cell death of human Fas-expressing WC8 cells in a dose-dependent manner, and the IC50 of 67 μg/mL.
Animal Protocol	Animal/Disease Models: Female balb/c (Bagg ALBino) mouse:[1] Doses: 10, 30, 100 or 300 mg/kg Route of Administration: Orally administered in a volume of 10 mL/kg Experimental Results: Inhibited the elevation of alanine aminotransferase and aspartate aminotransferase in a dose-dependent manner. Dramatically improved these parameters at doses of 100 mg/ kg and 300 mg/kg. Animal/Disease Models: Five-day-old SD rats[1] Doses: Route of Administration: Topically applied to the head and back of 5-day-old rats one time/day for 10 days Experimental Results: Alopecia scores Dramatically diminished throughout the entire observation period compared with the control group.
References	[1]. T Tsuda, et al. Inhibitory effect of M50054, a novel inhibitor of apoptosis, on anti-Fas-antibody-induced hepatitis and chemotherapy-induced alopecia. Eur J Pharmacol.2001 Dec 14;433(1):37-45.
Additional Infomation	2-[(2,6-dioxocyclohexyl)methyl]cyclohexane-1,3-dione is a beta-diketone.

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	4.2326 mL	21.1631 mL	42.3263 mL
	5 mM	0.8465 mL	4.2326 mL	8.4653 mL
	10 mM	0.4233 mL	2.1163 mL	4.2326 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.