Allosteric noncompetitive small molecule selective inhibitors of CD45 tyrosine phosphatase suppress T-cell receptor signals and inflammation in vivo.
CD45 is a receptor-like member of the protein tyrosine phosphatase (PTP) family. We screened in silico for small molecules binding at a predicted allosteric pocket unique to the CD45 intracellular domain, and validated inhibitors by in vitro phosphatase assays. Compound 211 exhibited a CD45 IC50 value of 200 nM and had >100-fold selectivity over six related PTPs. The relevance of the allosteric pocket was verified through site-directed mutagenesis. Compound 211 has a noncompetitive mechanism of action, and it is extremely effective at preventing dephosphorylation of substrate Lck phosphotyrosine (pY)-505 versus preventing dephosphorylation of Lck pY-393. In cultured primary T cells, compound 211 prevents T-cell receptor-mediated activation of Lck, Zap-70, and mitogen-activated protein kinase, and interleukin-2 production. In a delayed-type hypersensitivity reaction in vivo, compound 211 abolished inflammation. This work demonstrates a novel approach to develop effective allosteric inhibitors that can be expanded to target the corresponding allosteric domains of other receptor PTPs.
Lady Davis Institute-Jewish General Hospital (M.P., S.C., H.U.S.), Departments of Pharmacology and Therapeutics (M.P., H.U.S.), Biochemistry (I.A., E.P., M.L.T.), and Oncology (H.U.S.), Goodman Cancer Research Center (M.L.T.), and Segal Cancer Center, McGill University, Montreal, Quebec, Canada (H.U.S.); and Biotechnology Research Institute, National Research Council Canada, Montreal, Quebec, Canada (E.P.).
Michael D
MD
Perron
Shafinaz
S
Chowdhury
Isabelle
I
Aubry
Enrico
E
Purisima
Michel L
ML
Tremblay
H Uri
HU
Saragovi
D000494:Allosteric Regulation; D000495:Allosteric Site; D000818:Animals; D002478:Cells, Cultured; D004789:Enzyme Activation; D005260:Female; D006968:Hypersensitivity, Delayed / Q000188:drug therapy / Q000276:immunology; D007155:Immunologic Factors / Q000737:chemistry* / Q000494:pharmacology; D007166:Immunosuppressive Agents / Q000737:chemistry / Q000494:pharmacology; D007249:Inflammation / Q000188:drug therapy* / Q000276:immunology; D007376:Interleukin-2 / Q000096:biosynthesis; D017493:Leukocyte Common Antigens / Q000037:antagonists & inhibitors* / Q000235:genetics; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D062105:Molecular Docking Simulation; D016297:Mutagenesis, Site-Directed; D009285:Naphthoquinones / Q000737:chemistry / Q000494:pharmacology; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011948:Receptors, Antigen, T-Cell / Q000502:physiology*; D015398:Signal Transduction; D013329:Structure-Activity Relationship; D051746:ZAP-70 Protein-Tyrosine Kinase / Q000378:metabolism
D007155:Immunologic Factors; D007166:Immunosuppressive Agents; D007376:Interleukin-2; D009285:Naphthoquinones; D011948:Receptors, Antigen, T-Cell; D019000:Phosphotyrosine; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D051746:ZAP-70 Protein-Tyrosine Kinase; D020928:Mitogen-Activated Protein Kinases; D017493:Leukocyte Common Antigens