Vercirnon (GSK-1605786) sodium is an interface-active, selective CCR9 antagonist. Vercirnon sodium inhibits CCR9-mediated Ca2+ mobilization and chemotaxis on Molt-4 cells with IC50 values of 5.4 and 3.4 nM, respectively. Sodium Vercirnon is as selective for CCR9 as CCR1-12 and CX3CR1-7 (IC50s=>10 µM). Vercirnon sodium is similar in CCL25-directed engagement of the two spliced forms of CCR9 (CCR9A and CCR9B), with IC50 values of 2.8 and 2.6 nM, respectively.

Physicochemical Properties

Molecular Formula	C22H21CLN2NAO4S
Molecular Weight	466.913
Exact Mass	466.073
Elemental Analysis	C, 56.59; H, 4.32; Cl, 7.59; N, 6.00; Na, 4.92; O, 13.71; S, 6.87
CAS #	886214-18-2
Related CAS #	Vercirnon;698394-73-9
PubChem CID	71300757
Appearance	Light yellow to yellow solid powder
LogP	6.157
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	4
Heavy Atom Count	31
Complexity	862
Defined Atom Stereocenter Count	0
SMILES	CC(C)(C)C1=CC=C(C=C1)S(=O)(=O)/N=C/2\C=CC(=C\C2=C(/C3=CC=[N+](C=C3)[O-])\[O-])Cl.[Na+]
InChi Key	NKNHYMXTVRJZBC-YBZPVFAISA-M
InChi Code	InChI=1S/C22H21ClN2O4S.Na/c1-22(2,3)16-4-7-18(8-5-16)30(28,29)24-20-9-6-17(23)14-19(20)21(26)15-10-12-25(27)13-11-15;/h4-14,26H,1-3H3;/q;+1/p-1/b21-19-,24-20+;
Chemical Name	sodium;(Z)-[(6E)-6-(4-tert-butylphenyl)sulfonylimino-3-chlorocyclohexa-2,4-dien-1-ylidene]-(1-oxidopyridin-1-ium-4-yl)methanolate
Synonyms	Vercirnon sodium; 886214-18-2; Vercirnon sodium [USAN]; UNII-9NQF0M8R0M; 9NQF0M8R0M; GSK1605786A; GSK-1605786A; Benzenesulfonamide, N-(4-chloro-2-((1-oxido-4-pyridinyl)carbonyl)phenyl)-4-(1,1- dimethylethyl)-, sodium salt;
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 (e.g. under nitrogen), 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	CCR9B ( IC50 = 2.6 nM ); CCR9A ( IC50 = 2.8 nM )
ln Vitro	At an IC50 of 6.8 nM, vercirnon (GSK-1605786) sodium inhibits primary CCR9-expressing cells' chemotaxis toward CCL25. Human T cells cultured with retinoic acid (RA) exhibit chemotaxis induced by CCL25, which is inhibited by vercirnonodium. Vercirnonodium has an IC50 of 141 nM and inhibits CCL25-mediated chemotaxis of RA cultured cells in 100% human AB serum. Vercirnonodium, with IC50 values of 6.9 nM and 1.3 nM, respectively, is a strong inhibitor of CCL25-induced chemotaxis in mouse and rat thymocytes [1].
ln Vivo	Vercirnon (GSK1605786A) sodium (10, 50 mg/kg; subcutaneous injection; twice daily; starting at 2 weeks of age and up to 12 weeks of age) decreases the severity of intestinal inflammation in the TNFΔARE animal model [1].
Enzyme Assay	CCX282-B inhibited CCR9-mediated Ca(2+) mobilization and chemotaxis on Molt-4 cells with IC(50) values of 5.4 and 3.4 nM, respectively. In the presence of 100% human serum, CCX282-B inhibited CCR9-mediated chemotaxis with an IC(50) of 33 nM, and the addition of α1-acid glycoprotein did not affect its potency. CCX282-B inhibited chemotaxis of primary CCR9-expressing cells to CCL25 with an IC(50) of 6.8 nM. CCX282-B was an equipotent inhibitor of CCL25-directed chemotaxis of both splice forms of CCR9 (CCR9A and CCR9B) with IC(50) values of 2.8 and 2.6 nM, respectively. CCX282-B also inhibited mouse and rat CCR9-mediated chemotaxis[1].
Animal Protocol	Animal/Disease Models: C57BL/6 mice (TNFΔARE terminal ileitis mouse model) [1] Doses: 10, 50 mg/kg Route of Administration: subcutaneous injection; twice (two times) daily; starting at 2 weeks of age until 12 weeks of age Experimental Results: Severe inflammation associated with TNF overexpression was completely prevented at a dose of 50 mg/kg. Lower doses were similarly protective.
References	[1]. Characterization of CCX282-B, an orally bioavailable antagonist of the CCR9 chemokine receptor, for treatment of inflammatory bowel disease. J Pharmacol Exp Ther. 2010 Oct;335(1):61-9. [2]. CCR9 Antagonists in the Treatment of Ulcerative Colitis. Mediators Inflamm. 2015;2015:628340. [3]. Biarylsulfonamide CCR9 inhibitors for inflammatory bowel disease. Bioorg Med Chem Lett. 2015 Sep 1;25(17):3661-4.
Additional Infomation	Vercirnon is a novel, orally active anti-inflammatory agent that targets a chemokine receptor protein implicated in both Crohn's disease and ulcerative colitis, the two principal forms of IBD. It is under investigation in clinical trial NCT01611805 (Japanese Phase I of GSK1605786). Mechanism of Action Vercirnon, a small molecule, orally-available drug, is intended to control the inappropriate immune system response underlying IBD by blocking the activity of the CCR9 chemokine receptor. In adults, CCR9 is a highly specific receptor expressed by T cells that migrate selectively to the digestive tract. The trafficking of T cells to the small and large intestine causes persistent inflammation that may result in Crohn's disease or ulcerative colitis - the two principal forms of IBD. Inflammatory bowel disease, including Crohn's disease and ulcerative colitis, affects millions of people worldwide. CCR9 has been shown to be a key chemokine receptor mediating the local inflammatory responses in the GI tract. The CCR9 inhibitor Vercirnon advanced to phase 3 clinical trials, but carries several liabilities which we sought to improve.[3] While it has long been established that the chemokine receptor CCR9 and its ligand CCL25 are essential for the movement of leukocytes into the small intestine and the development of small-intestinal inflammation, the role of this chemokine-receptor pair in colonic inflammation is not clear. Toward this end, we compared colonic CCL25 protein levels in healthy individuals to those in patients with ulcerative colitis. In addition, we determined the effect of CCR9 pharmacological inhibition in the mdr1a(-/-) mouse model of ulcerative colitis. Colon samples from patients with ulcerative colitis had significantly higher levels of CCL25 protein compared to healthy controls, a finding mirrored in the mdr1a(-/-) mice. In the mdr1a(-/-) mice, CCR9 antagonists significantly decreased the extent of wasting and colonic remodeling and reduced the levels of inflammatory cytokines in the colon. These findings indicate that the CCR9:CCL25 pair plays a causative role in ulcerative colitis and suggest that CCR9 antagonists will provide a therapeutic benefit in patients with colonic inflammation.[2] The chemokine system represents a diverse group of G protein-coupled receptors responsible for orchestrating cell recruitment under both homeostatic and inflammatory conditions. Chemokine receptor 9 (CCR9) is a chemokine receptor known to be central for migration of immune cells into the intestine. Its only ligand, CCL25, is expressed at the mucosal surface of the intestine and is known to be elevated in intestinal inflammation. To date, there are no reports of small-molecule antagonists targeting CCR9. We report, for the first time, the discovery of a small molecule, CCX282-B, which is an orally bioavailable, selective, and potent antagonist of human CCR9. CCX282-B inhibited CCR9-mediated Ca(2+) mobilization and chemotaxis on Molt-4 cells with IC(50) values of 5.4 and 3.4 nM, respectively. In the presence of 100% human serum, CCX282-B inhibited CCR9-mediated chemotaxis with an IC(50) of 33 nM, and the addition of α1-acid glycoprotein did not affect its potency. CCX282-B inhibited chemotaxis of primary CCR9-expressing cells to CCL25 with an IC(50) of 6.8 nM. CCX282-B was an equipotent inhibitor of CCL25-directed chemotaxis of both splice forms of CCR9 (CCR9A and CCR9B) with IC(50) values of 2.8 and 2.6 nM, respectively. CCX282-B also inhibited mouse and rat CCR9-mediated chemotaxis. Inhibition of CCR9 with CCX282-B results in normalization of Crohn's disease such as histopathology associated with the TNF(ΔARE) mice. Analysis of the plasma level of drug associated with this improvement provides an understanding of the pharmacokinetic/pharmacodynamic relationship for CCR9 antagonists in the treatment of intestinal inflammation.[1]

Solubility Data

Solubility (In Vitro)

DMSO : ~100 mg/mL (~214.17 mM)

Solubility (In Vivo)

Solubility in Formulation 1: 5 mg/mL (10.71 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 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: ≥ 5 mg/mL (10.71 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 50.0 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.1417 mL	10.7087 mL	21.4174 mL
	5 mM	0.4283 mL	2.1417 mL	4.2835 mL
	10 mM	0.2142 mL	1.0709 mL	2.1417 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.