Tolimidone, formerly known as MLR-1023, is a novel and potent insulin receptor potentiator, and also a selective allosteric activator of Lyn kinase (EC50 of 63 nM).

Physicochemical Properties

Molecular Formula	C11H10N2O2
Molecular Weight	202.2093
Exact Mass	202.074
Elemental Analysis	C, 65.34; H, 4.98; N, 13.85; O, 15.82
CAS #	41964-07-2
Related CAS #	41964-07-2
PubChem CID	39065
Appearance	White to yellow solid powder
Density	1.2±0.1 g/cm3
Melting Point	163-164ºC
Index of Refraction	1.598
LogP	1.37
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	2
Rotatable Bond Count	2
Heavy Atom Count	15
Complexity	310
Defined Atom Stereocenter Count	0
SMILES	O=C1N=CC(OC2=CC=CC(C)=C2)=CN1
InChi Key	HJQILFPVRNHTIG-UHFFFAOYSA-N
InChi Code	InChI=1S/C11H10N2O2/c1-8-3-2-4-9(5-8)15-10-6-12-11(14)13-7-10/h2-7H,1H3,(H,12,13,14)
Chemical Name	5-(3-methylphenoxy)-1H-pyrimidin-2-one
Synonyms	CP 26,154; CP 26,154; CP 26,154; NSC 314335; MLR-1023; MLR 1023; MLR1023; 41964-07-2
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	Insulin Receptor; Lyn (EC50 = 63 nM)
ln Vitro	Tolimidone (MLR-1023) exhibits a 50% increase in enzyme activity that is repeatable when incubated with Lyn kinase. Lyn kinase activation is increased in response to tolimidone in a concentration-dependent manner, increasing by 2.3 and 2.1 fold at concentrations of 3 and 10 μM, respectively. When Tolimidone (100 μM) is added, Lyn kinase activity increases threefold at every tested ATP concentration (Vmax=2601 U/mg). The length of the preincubation period in the absence of ATP determines how much Lyn kinase is activated by tolimidone[1].
ln Vivo	Tolimidone (MLR-1023) (30 mg/kg i.p.) is administered, blood glucose levels are significantly (p<0.05) lowered to 148 and 158 mg/dL, respectively, 30 and 90 minutes later. Adiponectin production and adipocyte differentiation are both markedly increased by tolimidone, increasing them by 19 and 3.7 times, respectively[1]. A maximal effect of 30 mg/kg is observed when tolimidone is administered, eliciting a dose-dependent potentiation of the insulin response[2].
Enzyme Assay	Tolimidone (MLR-1023) (10 μM) is preincubated with kinase and fluoroscein-labeled protein substrate for every kinase test. The amount of fluoroscein phosphopeptide is measured after the addition of ATP, at a concentration equal to or less than the Km for each kinase, starts the reaction. There are two assays performed[1].
Cell Assay	In mouse 3T3-L1 cells, adipocyte differentiation is evaluated following an 8-day incubation period with either Tolimidone (MLR-1023) or Rosiglitazone (10 μM). Transiently transfected cells containing the appropriate DNA constructs (pGAL4/PPARα, σ, or γ) cotransfected with a luciferase reporter vector are used for PPAR (α, σ, and γ) transactivation studies. For twenty-four hours, transfected cells are incubated with tolimidone or a suitable reference compound. The level of PPARα, σ, and γ activation is measured by luciferase activity[1].
Animal Protocol	Baseline glucose, glucose tolerance, and insulin levels are studied in male mice aged 8 to 10 weeks. Intraperitoneal injections of tolimidone (MLR-1023) are given at dose volumes ranging from 5 to 10 mL/kg. A tail snip is used to obtain 5 μL of blood, which is then immediately applied to a glucose test strip. Liquid chromatography/tandem mass spectrometry is used to measure the levels of Tolimidone in blood. The levels are then compared to a standard curve of Tolimidone prepared in blood[1].
References	[1]. MLR-1023 is a potent and selective allosteric activator of Lyn kinase in vitro that improves glucose tolerance in vivo. J Pharmacol Exp Ther. 2012 Jul;342(1):15-22. [2]. The Lyn kinase activator MLR-1023 is a novel insulin receptor potentiator that elicits a rapid-onset and durable improvement in glucose homeostasis in animal models of type 2 diabetes. J Pharmacol Exp Ther. 2012 Jul;342(1):23-32.

Solubility Data

Solubility (In Vitro)

DMSO: 40~150 mg/mL (197.8~741.8 mM) Ethanol: ~23 mg/mL (~113.7 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (12.36 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (12.36 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: 2.5 mg/mL (12.36 mM) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 4: 5%dmso+95%corn oil: 5mg/mL  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	4.9454 mL	24.7268 mL	49.4535 mL
	5 mM	0.9891 mL	4.9454 mL	9.8907 mL
	10 mM	0.4945 mL	2.4727 mL	4.9454 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.