Mechanism of action and efficacy of RX-111, a thieno[2,3-c]pyridine derivative and small molecule inhibitor of protein interaction with glycosaminoglycans (SMIGs), in delayed-type hypersensitivity, TNBS-induced colitis and experimental autoimmune encephalomyelitis
Objective and design
Elucidate the mechanism of action of the small molecule inhibitor of protein binding to glycosaminoglycans, RX-111 and assay its anti-inflammatory activity in animal models of inflammatory disease.
The glycosaminoglycan, heparin, was used in the mechanism of action study of RX-111. Human T lymphocytes and umbilical vein endothelial cells were used to assay the in vitro activity of RX-111. Mouse and rat models of disease were used to assay the anti-inflammatory activity of RX-111 in vivo.
Circular dichroism and UV/Vis absorption spectroscopy were used to study the binding of RX-111 to the glycosaminoglycan, heparin. T lymphocyte rolling on endothelial cells under shear flow was used to assay RX-111 activity in vitro. Delayed-type hypersensitivity (DTH) and tri-nitrobenzene sulfonic acid (TNBS)-induced colitis in mice and experimental autoimmune encephalomyelitis (EAE) in rats were used to assay anti-inflammatory activity of RX-111 in vivo.
RX-111 was shown to bind directly to heparin. It inhibited leukocyte rolling on endothelial cells under shear flow and reduced inflammation in the mouse model of DTH. RX-111 was efficacious in the mouse model of inflammatory bowel disease, TNBS-induced colitis and the rat model of multiple sclerosis, EAE.
RX-111 exercises its broad spectrum anti-inflammatory activity by a singular mechanism of action, inhibition of protein binding to the cell surface GAG, heparan sulfate. RX-111 and related thieno[2,3-c]pyridine derivatives are potential therapeutics for the treatment of inflammatory and autoimmune diseases.
KeywordsSmall molecule drug Glycosaminoglycan Heparin binding protein Heparan sulfate Inflammation Autoimmune disease
- SMIG or SMIGs
Small molecule inhibitors of protein interaction with glycosaminoglycans
High endothelium venules
Human umbilical vein endothelial
Experimental autoimmune encephalomyelitis
Myelin basic protein
Myelin oligodendrocyte glycoprotein
Mucosal addressin cell adhesion molecule 1
Peripheral node addressins
Progressive multifocal leukoencephalopathy
The research was supported by grants (37373, 39243) awarded by the Office of The Chief Scientist, Israel. We are grateful to Mr. Ra’anan Margalit and the late Dr. Valentin Grabovsky for technical assistance and professional advice.
Supplementary material 2 (AVI 1995 kb)
Supplementary material 3 (AVI 2085 kb)
- 14.Noravyan AS, et al. Synthesis of 2-and 4-substituted 5,6,7,8-tetrahydro-7-isopropylpyrido[4′,3′:4,5]thieno[2,3-d]pyrimidines. Pharm Chem J. 1980;14:111–3.Google Scholar
- 26.Prankerd RJ. Critical compilation of pK a values for pharmaceutical substances. In: Brittain HG, editor. Profiles of drug substances, excipients and related methodology, vol. 33; 2007.Google Scholar
- 54.Kansas GS. Selectins and their ligands: current concepts and controversies. Blood. 1966;88:3259–87.Google Scholar
- 55.Watson SR. Glycoprotein ligands for L-selectin. In: Vestweber D, editor. The selectins. Amsterdam: Harwood Academic Publishers; 1997. p. 179–93.Google Scholar