MK0812 Succinate (MK-0812) is a novel, potent and selective small molecule CCR2 antagonist with anti-Inflammatory and immunomodulatory activity. MK0812 caused an increase in the CCR2 ligand CCL2 and a selective decrease in the frequency of peripheral blood monocytes. The therapeutic potential for targeting CXCR2/CXCR1 in human arthritides is highlighted by the significantly greater impact of CXCR2/CXCR1 antagonism in this model of arthritis compared to CCR2 antagonism. In the tissue and synovial fluid of rheumatoid arthritis patients, neutrophils and monocytes are highly prevalent. It was investigated how small molecule chemokine receptor antagonists, like MK-0812, might prevent these cells from migrating in arthritis caused by anti-collagen antibodies.

Physicochemical Properties

Molecular Formula	C28H40F3N3O7
Molecular Weight	587.628318786621
Exact Mass	587.28
Elemental Analysis	C, 57.23; H, 6.86; F, 9.70; N, 7.15; O, 19.06
CAS #	851916-42-2
Related CAS #	MK-0812; 624733-88-6
PubChem CID	60196412
Appearance	White to off-white solid powder
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	11
Rotatable Bond Count	7
Heavy Atom Count	41
Complexity	793
Defined Atom Stereocenter Count	4
SMILES	CC(C)[C@@]1(CC[C@H](C1)[NH2+][C@H]2CCOC[C@H]2OC)C(=O)N3CCC4=C(C3)C=C(C=N4)C(F)(F)F.C(CC(=O)[O-])C(=O)O
InChi Key	WPPJJJUIDYHHSM-PXUYIWLPSA-N
InChi Code	InChI=1S/C24H34F3N3O3.C4H6O4/c1-15(2)23(7-4-18(11-23)29-20-6-9-33-14-21(20)32-3)22(31)30-8-5-19-16(13-30)10-17(12-28-19)24(25,26)27;5-3(6)1-2-4(7)8/h10,12,15,18,20-21,29H,4-9,11,13-14H2,1-3H3;1-2H2,(H,5,6)(H,7,8)/t18-,20+,21-,23+;/m1./s1
Chemical Name	butanedioic acid;[(1S,3R)-3-[[(3S,4S)-3-methoxyoxan-4-yl]amino]-1-propan-2-ylcyclopentyl]-[3-(trifluoromethyl)-7,8-dihydro-5H-1,6-naphthyridin-6-yl]methanone
Synonyms	MK-0812 Succinate; MK0812 Succinate; MK-0812; MK 0812 Succinate
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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.
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	CCR2
ln Vitro	MK-0812 is a potent and selective antagonist of CCR2 [1]. MK-0812 exhibits concentration-dependent partial inhibition of all MCP-1-mediated responses, with an IC50 of 3.2 nM. This amount is comparable to the potency found in MK-0812's suppression of 125I-MCP-1 binding on isolated monocytes (IC50 4.5 nM). MK-0812, in fact, not only completely prevents the exogenous MCP-1-induced shape change response, but it also causes a monocyte forward scatter measurement that is lower than basal or unstimulated levels. MK-0812 also prevents MCP-1-induced monocyte morphological change when added to rhesus blood. In whole blood assays, the IC50 for MK-0812 is 8 nM[2].
ln Vivo	MK-0812 (30 mg/kg, p.o.) decreases the frequency of Ly6G-Ly6Chi monocytes in peripheral blood, but has no effect on the frequency of Ly6G+Ly6C+ neutrophils in circulation. Additionally, administration of MK-0812 results in an increase in the CCR2 ligand CCL2 and a dose-dependent decrease in circulating Ly6Chi monocytes[1]. To keep the amount of MK-0812 in the blood at a steady level, it is continuously infused intravenously[2].
Animal Protocol	Mice: The mice used are 8–10 week old female BALB/c mice. SCH563705 or MK-0812 are given orally at a dose of 30 mg/kg using a 0.4% methylcellulose (MC) solution (p.o.). Using flow cytometry, the frequency of CD11b+Ly6G+Ly6C+ neutrophils and CD11b+Ly6G+Ly6Chi monocytes is ascertained two hours later[1].
References	[1]. Pharmacological targeting reveals distinct roles for CXCR2/CXCR1 and CCR2 in a mouse model of arthritis.Biochem Biophys Res Commun. 2010 Jan 1;391(1):1080-6. [2]. Assessment of chemokine receptor function on monocytes in whole blood: In vitro and ex vivo evaluations of a CCR2 antagonist.J Immunol Methods. 2010 Jan 31;352(1-2):101-10.

Solubility Data

Solubility (In Vitro)

DMSO: ≥ 32 mg/mL (~54.5 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.25 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 (4.25 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 (4.25 mM) (saturation unknown) 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.7018 mL	8.5088 mL	17.0175 mL
	5 mM	0.3404 mL	1.7018 mL	3.4035 mL
	10 mM	0.1702 mL	0.8509 mL	1.7018 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.