ABSTRACT
The design and synthesis of a novel class of 7-azaspiro[3.5]nonane GPR119 agonists are described. In this series, optimization of the right piperidine N-capping group (R2) and the left aryl group (R3) led to the identification of compound 54g as a potent GPR119 agonist. Compound 54g showed a desirable PK profile in Sprague-Dawley (SD) rats and a favorable glucose lowering effect in diabetic rats.
Subject(s)
Alkanes/chemistry , Drug Design , Receptors, G-Protein-Coupled/agonists , Alkanes/chemical synthesis , Alkanes/pharmacokinetics , Animals , Blood Glucose/analysis , Cell Line , Diabetes Mellitus, Experimental/drug therapy , Diabetes Mellitus, Experimental/pathology , Glucose Tolerance Test , Half-Life , Humans , Hypoglycemic Agents/chemical synthesis , Hypoglycemic Agents/chemistry , Hypoglycemic Agents/therapeutic use , Microsomes, Liver/metabolism , Piperidines/chemistry , Rats , Rats, Sprague-Dawley , Receptors, G-Protein-Coupled/metabolism , Structure-Activity RelationshipABSTRACT
We previously reported a novel series of 1H-pyrazolo[3,4-c]pyridine derivatives and the identification of compound 4b as a highly potent GPR119 agonist. However, the advancement of preclinical evaluations of compound 4b is expected to be difficult because of the compound's significantly poor aqueous solubility (0.71µM at pH6.8). In this article, we describe the further optimization of compound 4b focusing on the improvement of its aqueous solubility. Optimization of the central spacer, left-hand aryl group and right-hand piperidine N-capping group led to the identification of a potent GPR119 agonist, 3H-[1,2,3]triazolo[4,5-c]pyridine derivative 32o, with improved solubility (15.9µM at pH6.8).
Subject(s)
Pyridines/pharmacology , Receptors, G-Protein-Coupled/antagonists & inhibitors , Triazoles/pharmacology , Dose-Response Relationship, Drug , HEK293 Cells , Humans , Molecular Structure , Pyridines/chemical synthesis , Pyridines/chemistry , Structure-Activity Relationship , Triazoles/chemical synthesis , Triazoles/chemistryABSTRACT
Design and synthesis of a novel class of 1H-pyrazolo[3,4-c]pyridine GPR119 receptor agonists are described. Lead compound 4 was identified through the ligand-based drug design approach. Modification of the left-hand aryl group (R(1)) and right-hand piperidine N-capping group (R(2)) led to the identification of compound 24 as a single-digit nanomolar GPR119 agonist.
Subject(s)
Drug Design , Pyridines/pharmacology , Receptors, G-Protein-Coupled/agonists , Dose-Response Relationship, Drug , Humans , Molecular Structure , Pyridines/chemical synthesis , Pyridines/chemistry , Structure-Activity RelationshipABSTRACT
We isolated the cDNA of a gene, designated smooth muscle-associated protein 8 (smap8), during a search for new genes expressed in human aortic smooth muscle cells. The full-length smap8 cDNA is 3241 bp long and contains an open reading frame of 1113 bp encoding an approximately 45 kDa soluble protein identical to NDRG4 protein. Smap8 mRNA was expressed predominantly in the brain and heart, and moderately in vascular smooth muscle cells. Expression of smap8 mRNA was induced within 3-12 h by treatment with 10 mm homocysteine in rat aortic smooth muscle cells (A10 cells). Expression of exogenous smap8 markedly reduced both the proliferation and migration rates of rat A10 cells, however, PDGF-induced proliferation was significantly enhanced in smap8-expressed cells compared with mock-transfected cells. To ascertain the involvement of smap8 in mitogenesis, we tested the effects of stimulation of smap8, MEK1/2 or ERK1/2, which is known as a proliferation relating intermediate, by various growth factors and cytokines. PDGF was the most prominent in promoting phosphorylation of the smap8 protein. PDGF-dependent phosphorylation of smap8 was induced prior to ERK1/2 activation, and was repressed by staurosporine, a general inhibitor of serine/threonine kinases. Furthermore, activation of both MEK1/2 and ERK1/2 was markedly enhanced in these cells. Smap8 might therefore regulate the potentiation of ERK1/2 signalling induced by PDGF treatment. Our results imply that smap8 is involved in the regulation of mitogenic signalling in vascular smooth muscle cells, possibly in response to a homocysteine-induced injury.
Subject(s)
Homocysteine/chemistry , Muscle Proteins/chemistry , Muscle Proteins/physiology , Muscle, Smooth, Vascular/cytology , Nerve Tissue Proteins , Amino Acid Sequence , Animals , Blotting, Northern , Cell Division , Cell Line , Cell Movement , Cell Nucleus/metabolism , Cells, Cultured , Cloning, Molecular , DNA, Complementary/metabolism , Drosophila melanogaster/metabolism , Gene Expression Regulation , Homocysteine/metabolism , Humans , Immunoblotting , MAP Kinase Kinase 1 , MAP Kinase Kinase 2 , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3 , Mitogen-Activated Protein Kinase Kinases/metabolism , Mitogen-Activated Protein Kinases/metabolism , Molecular Sequence Data , Muscle, Smooth/cytology , Peptides/chemistry , Phosphorylation , Platelet-Derived Growth Factor/metabolism , Protein Serine-Threonine Kinases/metabolism , Protein Structure, Tertiary , Protein-Tyrosine Kinases/metabolism , RNA/metabolism , RNA, Messenger/metabolism , Rats , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Amino Acid , Signal Transduction , Time Factors , Tissue Distribution , TransfectionABSTRACT
Proliferation of aortic smooth muscle cells is an important event in vascular lesion formation. To identify new genes that are involved in neointima formation, we constructed an aortic 3'-directed cDNA library. The novel cDNA of a gene designated smooth muscle associated protein 2 (smap2) was isolated. The full-length cDNA of smap2 is 2914 base pairs long and contains an open reading frame of 1338 base pairs. Dot blot analysis revealed that smap2 was expressed particularly in aorta. The deduced amino acid sequence of smap2 contains two thyroglobulin type-1 domains, two EF-hand calcium-binding domains and putative signal peptide. Furthermore, we demonstrated that smap2 mRNA was upregulated during neointima formation in a rat carotid endarterectomy model. These findings suggest that smap2 might be involved in the progression of atherosclerosis in aorta.
