ABSTRACT
The discovery of a novel series of S1P1 agonists is described. Starting from a micromolar HTS positive, iterative optimization gave rise to several single-digit nanomolar S1P1 agonists. The compounds were able to induce internalization of the S1P1 receptor, and a selected compound was shown to be able to induce lymphopenia in mice after oral dosing.
Subject(s)
Antineoplastic Agents/chemistry , Receptors, Lysosphingolipid/agonists , Administration, Oral , Animals , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/pharmacokinetics , Cell Line, Tumor , Drug Discovery , Fingolimod Hydrochloride , High-Throughput Screening Assays , Humans , Mice , Microsomes, Liver/metabolism , Propylene Glycols/chemistry , Propylene Glycols/pharmacology , Rats , Receptors, Lysosphingolipid/metabolism , Sphingosine/analogs & derivatives , Sphingosine/chemistry , Sphingosine/pharmacology , Structure-Activity RelationshipABSTRACT
Inadequate remyelination of brain white matter lesions has been associated with a failure of oligodendrocyte precursors to differentiate into mature, myelin-producing cells. In order to better understand which genes play a critical role in oligodendrocyte differentiation, we performed time-dependent, genome-wide gene expression studies of mouse Oli-neu cells as they differentiate into process-forming and myelin basic protein-producing cells, following treatment with three different agents. Our data indicate that different inducers activate distinct pathways that ultimately converge into the completely differentiated state, where regulated gene sets overlap maximally. In order to also gain insight into the functional role of genes that are regulated in this process, we silenced 88 of these genes using small interfering RNA and identified multiple repressors of spontaneous differentiation of Oli-neu, most of which were confirmed in rat primary oligodendrocyte precursors cells. Among these repressors were CNP, a well-known myelin constituent, and three phosphatases, each known to negatively control mitogen-activated protein kinase cascades. We show that a novel inhibitor for one of the identified genes, dual-specificity phosphatase DUSP10/MKP5, was also capable of inducing oligodendrocyte differentiation in primary oligodendrocyte precursors. Oligodendrocytic differentiation feedback loops may therefore yield pharmacological targets to treat disease related to dysfunctional myelin deposition.
Subject(s)
Cell Differentiation/physiology , Gene Regulatory Networks , Oligodendroglia/physiology , Signal Transduction/physiology , Animals , Cell Differentiation/drug effects , Cells, Cultured , Colforsin/pharmacology , Dexamethasone/pharmacology , Dual-Specificity Phosphatases/antagonists & inhibitors , Dual-Specificity Phosphatases/physiology , Gene Silencing , Genome-Wide Association Study , Mice , Myelin Basic Protein/biosynthesis , Neurogenesis/physiology , Oligodendroglia/cytology , Rats , Signal Transduction/drug effects , Tretinoin/pharmacologyABSTRACT
Class I phosphoinositide 3-kinases (PI3Ks), in particular PI3Kgamma, have become attractive drug targets for inflammatory and autoimmune diseases. Here, we disclose a novel series of furan-2-ylmethylene thiazolidinediones as selective, ATP-competitive PI3Kgamma inhibitors. Structure-based design and X-ray crystallography of complexes formed by inhibitors bound to PI3Kgamma identified key pharmacophore features for potency and selectivity. An acidic NH group on the thiazolidinedione moiety and a hydroxy group on the furan-2-yl-phenyl part of the molecule play crucial roles in binding to PI3K and contribute to class IB PI3K selectivity. Compound 26 (AS-252424), a potent and selective small-molecule PI3Kgamma inhibitor emerging from these efforts, was further profiled in three different cellular PI3K assays and shown to be selective for class IB PI3K-mediated cellular effects. Oral administration of 26 in a mouse model of acute peritonitis led to a significant reduction of leukocyte recruitment.
Subject(s)
Furans/chemical synthesis , Phosphoinositide-3 Kinase Inhibitors , Thiazolidinediones/chemical synthesis , Acute Disease , Animals , Bone Marrow Cells/drug effects , Bone Marrow Cells/physiology , Cells, Cultured , Chemotaxis/drug effects , Class Ib Phosphatidylinositol 3-Kinase , Crystallography, X-Ray , Furans/chemistry , Furans/pharmacology , Humans , Isoenzymes/antagonists & inhibitors , Isoenzymes/chemistry , Mast Cells/drug effects , Mast Cells/metabolism , Mice , Models, Molecular , Molecular Structure , Monocytes/drug effects , Monocytes/physiology , Neutrophils/immunology , Peritonitis/chemically induced , Peritonitis/drug therapy , Peritonitis/immunology , Phosphatidylinositol 3-Kinases/chemistry , Phosphorylation , Proto-Oncogene Proteins c-akt/metabolism , Structure-Activity Relationship , Thiazolidinediones/chemistry , Thiazolidinediones/pharmacology , ThioglycolatesABSTRACT
Phosphoinositide 3-kinases (PI3K) have long been considered promising drug targets for the treatment of inflammatory and autoimmune disorders as well as cancer and cardiovascular diseases. But the lack of specificity, isoform selectivity and poor biopharmaceutical profile of PI3K inhibitors have so far hampered rigorous disease-relevant target validation. Here we describe the identification and development of specific, selective and orally active small-molecule inhibitors of PI3Kgamma (encoded by Pik3cg). We show that Pik3cg(-/-) mice are largely protected in mouse models of rheumatoid arthritis; this protection correlates with defective neutrophil migration, further validating PI3Kgamma as a therapeutic target. We also describe that oral treatment with a PI3Kgamma inhibitor suppresses the progression of joint inflammation and damage in two distinct mouse models of rheumatoid arthritis, reproducing the protective effects shown by Pik3cg(-/-) mice. Our results identify selective PI3Kgamma inhibitors as potential therapeutic molecules for the treatment of chronic inflammatory disorders such as rheumatoid arthritis.
