ABSTRACT
We herein describe the results of further evolution of glycogen synthase kinase (GSK)-3ß inhibitors from our promising compounds containing a 3-methylmorpholine moiety. Transformation of the morpholine moiety into a piperazine moiety resulted in potent GSK-3ß inhibitors. SAR studies focused on the nitrogen atom of the piperazine moiety revealed that a phenyl group afforded potent inhibitory activity toward GSK-3ß. Docking studies indicated that the phenyl group on the piperazine nitrogen atom and the methyl group on the piperazine make cation-π and CH-π interactions with GSK-3ß respectively. 4-Methoxyphenyl analogue 29 showed most potent inhibitory activity toward GSK-3ß with good in vitro and in vivo pharmacokinetic profiles, and 29 demonstrated a significant decrease in tau phosphorylation after oral administration in mice.
Subject(s)
Drug Discovery , Protein Kinase Inhibitors/pharmacology , Pyrimidinones/pharmacology , Dose-Response Relationship, Drug , Glycogen Synthase Kinase 3 beta/metabolism , Humans , Molecular Docking Simulation , Molecular Structure , Protein Kinase Inhibitors/chemical synthesis , Protein Kinase Inhibitors/chemistry , Pyrimidinones/chemical synthesis , Pyrimidinones/chemistry , Structure-Activity RelationshipABSTRACT
We herein describe the results of further evolution of glycogen synthase kinase (GSK)-3ß inhibitors from our promising compounds containing a 2-phenylmorpholine moiety. Transformation of the morpholine moiety into a piperazine moiety resulted in potent GSK-3ß inhibitors. SAR studies focused on the phenyl moiety revealed that a 4-fluoro-2-methoxy group afforded potent inhibitory activity toward GSK-3ß. Based on docking studies, new hydrogen bonding between the nitrogen atom of the piperazine moiety and the oxygen atom of the main chain of Gln185 has been indicated, which may contribute to increased activity compared with that of the corresponding phenylmorpholine analogues. Effect of the stereochemistry of the phenylpiperazine moiety is also discussed.
Subject(s)
Drug Discovery , Piperazines/pharmacology , Protein Kinase Inhibitors/pharmacology , Pyrimidinones/pharmacology , Dose-Response Relationship, Drug , Glycogen Synthase Kinase 3 beta/metabolism , Humans , Molecular Docking Simulation , Molecular Structure , Piperazines/chemical synthesis , Piperazines/chemistry , Protein Kinase Inhibitors/chemical synthesis , Protein Kinase Inhibitors/chemistry , Pyrimidinones/chemical synthesis , Pyrimidinones/chemistry , Structure-Activity RelationshipABSTRACT
Laparoscopic cholecystectomy has become the gold standard for the treatment of cholelithiasis, and many reports of single-incision laparoscopic cholecystectomy have been published in the past few years. Situs inversus totalis is a very rare condition, but the variant anatomy should not preclude a minimally invasive approach to surgery. We report a case of successful single-port laparoscopic cholecystectomy in a patient with situs inversus totalis, describe the technical advantages, and review the literature.
Subject(s)
Cholecystectomy, Laparoscopic/methods , Cholelithiasis/surgery , Situs Inversus/complications , Aged , Cholecystectomy, Laparoscopic/instrumentation , Cholelithiasis/complications , Cholelithiasis/diagnosis , Humans , MaleABSTRACT
We herein describe the results of further evolution of GSK-3ß inhibitors for Alzheimer's disease from our promising compounds with in vivo tau phosphorylation inhibitory activity by oral administration. Introduction of a low alkyl group instead of the phenyl group at the 3-position of the morpholine moiety aiming to improve pharmacokinetic profiles resulted in potent low molecular weight GSK-3ß inhibitors with good in vitro pharmacokinetic profiles, which also showed in vivo tau phosphorylation inhibitory activity by oral administration. Effect of the stereochemistry of the alkyl moiety is also discussed using docking models.
Subject(s)
Alzheimer Disease/enzymology , Glycogen Synthase Kinase 3/antagonists & inhibitors , Morpholines/chemistry , Morpholines/pharmacology , Pyrimidines/chemistry , Pyrimidines/pharmacology , Administration, Oral , Alzheimer Disease/drug therapy , Alzheimer Disease/metabolism , Animals , Drug Discovery , Glycogen Synthase Kinase 3/metabolism , Glycogen Synthase Kinase 3 beta , Humans , Mice , Molecular Docking Simulation , Morpholines/administration & dosage , Morpholines/pharmacokinetics , Phosphorylation/drug effects , Pyrimidines/administration & dosage , Pyrimidines/pharmacokinetics , tau Proteins/metabolismABSTRACT
A series of 2-(2-phenylmorpholin-4-yl)pyrimidin-4(3H)-ones was synthesized and examined for their inhibitory activity against glycogen synthase kinase-3ß (GSK-3ß). We found 21, 29 and 30 to possess potent in vitro GSK-3ß inhibitory activity with good in vitro PK profiles. 21 demonstrated significant decrease of tau phosphorylation after oral administration in mice and excellent PK profiles.
Subject(s)
Glycogen Synthase Kinase 3/antagonists & inhibitors , Morpholines/chemical synthesis , Morpholines/pharmacology , Pyrimidinones/chemistry , Pyrimidinones/pharmacology , Administration, Oral , Animals , Binding Sites , Cytochrome P-450 Enzyme Inhibitors , Cytochrome P-450 Enzyme System/metabolism , Enzyme Activation/drug effects , Female , Glycogen Synthase Kinase 3/metabolism , Glycogen Synthase Kinase 3 beta , Half-Life , Humans , Male , Mice , Microsomes, Liver/drug effects , Microsomes, Liver/metabolism , Molecular Docking Simulation , Morpholines/pharmacokinetics , Phosphorylation/drug effects , Protein Kinase Inhibitors/chemical synthesis , Protein Kinase Inhibitors/chemistry , Protein Kinase Inhibitors/pharmacokinetics , Protein Kinase Inhibitors/pharmacology , Protein Structure, Tertiary , Pyrimidinones/chemical synthesis , Pyrimidinones/pharmacokinetics , Rats , Rats, Sprague-Dawley , Stereoisomerism , tau Proteins/metabolismABSTRACT
The discovery of a series of 6-(4-pyridyl)pyrimidin-4(3H)-ones derived from a hit compound with low molecular weight and sufficient chemical space is reported. Transformation of substituents led to subnanomolar potent inhibitors with in vivo tau phoshorylation lowering activity.
Subject(s)
Glycogen Synthase Kinase 3/antagonists & inhibitors , Protein Kinase Inhibitors/chemistry , Pyrimidinones/chemistry , Binding Sites , Enzyme Activation , Glycogen Synthase Kinase 3/metabolism , Glycogen Synthase Kinase 3 beta , Molecular Docking Simulation , Protein Binding/drug effects , Protein Kinase Inhibitors/chemical synthesis , Protein Kinase Inhibitors/metabolism , Protein Kinase Inhibitors/pharmacology , Protein Structure, Tertiary , Pyrimidinones/chemical synthesis , Pyrimidinones/metabolism , Pyrimidinones/pharmacology , Structure-Activity RelationshipABSTRACT
BACKGROUND: Dendritic cells (DCs) play a critical role in the immune response. The aim of this study was to investigate apoptotic tumor cells as an antigen source for DC maturation. MATERIALS AND METHODS: We compared the efficacy of ultraviolet (UV)-irradiation with that of gamma-irradiation in the induction of apoptosis of tumor cells. Phenotypic and functional changes of DCs were analyzed after co-incubation with UV-irradiated tumor cells. RESULTS: UV-irradiation (1.8 J/cm2) was a more reliable method of inducing apoptosis than gamma-irradiation (20,000 rad). The expression of costimulatory molecules on DCs was upregulated after co-incubation with UV-irradiated tumor cells. When we performed allogeneic mixed lymphocyte reaction assay, UV-irradiated tumor cells-pulsed DCs stimulated allogeneic T lymphocytes more efficiently than DCs pulsed with gamma-irradiated cells (HT29, p = 0.0419 and WiDr, p = 0.0076). CONCLUSION: UV-irradiation reliably induces apoptosis in cultured colorectal cancer cells. DCs mature in function and expression of costimulatory molecules after co-incubation with apoptotic tumor cells. The clinical implications warrant further study.
Subject(s)
Colorectal Neoplasms/immunology , Dendritic Cells/immunology , Antigens, CD/biosynthesis , Antigens, CD/immunology , Apoptosis/radiation effects , B7-1 Antigen/biosynthesis , B7-1 Antigen/immunology , B7-2 Antigen , Cell Communication/immunology , Colorectal Neoplasms/pathology , Dendritic Cells/cytology , HT29 Cells , Humans , Lymphocyte Culture Test, Mixed , Membrane Glycoproteins/biosynthesis , Membrane Glycoproteins/immunology , Tumor Cells, Cultured , Ultraviolet Rays , Up-RegulationABSTRACT
Cytokine gene therapy is applied in clinical studies of tumors, and IFN-alpha and IL-12 are widely used for cancer immunotherapy. Using a poorly immunogenic murine colorectal cancer cell line, MC38, we compared antitumor effects of IFN-alpha and IL-12. Transduced MC38 cell lines expressing IFN-alpha or IL-12 (MC38-IFNalpha or MC38-IL12, respectively) were established using retroviral vectors. Transduction of IFN-alpha or IL-12 gene to MC38 cells significantly reduced tumorigenicity in immunocompetent mice. When tumor-free mice initially injected with MC38-IFNalpha or MC38-IL12 cells were reinjected contralaterally with wild-type MC38 cells (MC38-WT) after 35 days, 7 of 12 or 2 of 12 mice rejected MC38-WT cells, respectively. In therapy-model mice with established tumor derived from MC38-WT cells, inoculation of gene-transduced cells significantly suppressed growth of the tumor in MC38-IFNalpha-inoculated groups, but not in the IL-12-inoculated group. Immunohistologic and flow cytometric analyses showed marked infiltration of CD8(+) cells in wild-type tumors of mice inoculated with IFN-alpha-expressing cells. Leukocyte-depletion experiments implicated CD8(+) T cells in tumor rejection induced by IFN-alpha-transduction; both CD8(+) T cells and natural killer cells were implicated in the more modest antitumor effect from IL-12 expression. To investigate induction of tumor-specific immune responses, we stimulated splenocytes from tumor-free mice twice in vitro with genetically modified MC38 cells. In vitro stimulations with MC38-IFNalpha cells induced definite MC38-specific lysis, but not stimulations with MC38-IL-12 cells. Injecting combination of MC38-IFNalpha and MC38-IL-12 cells caused an additive antitumor effect in the therapy model. These data suggested that IFN-alpha induces cytotoxic T lymphocytes and elicits long-lasting tumor-specific immunity, whereas IL-12 seems to stimulate non-specific killing. With additional refinements, combined IFN-alpha and IL-12 gene therapy might warrant clinical trials.
