ABSTRACT
We report herein the identification of MK-4409, a potent and selective fatty acid amide hydrolase (FAAH) inhibitor. Starting from a high throughput screening (HTS) hit, medicinal chemistry efforts focused on optimizing of FAAH inhibition in vitro potency, improving the pharmacokinetic (PK) profile, and increasing in vivo efficacy in rodent inflammatory and neuropathic pain assays.
ABSTRACT
We report herein the discovery of a fatty acid amide hydrolase (FAAH) positron emission tomography (PET) tracer. Starting from a pyrazole lead, medicinal chemistry efforts directed toward reducing lipophilicity led to the synthesis of a series of imidazole analogues. Compound 6 was chosen for further profiling due to its appropriate physical chemical properties and excellent FAAH inhibition potency across species. [(11)C]-6 (MK-3168) exhibited good brain uptake and FAAH-specific signal in rhesus monkeys and is a suitable PET tracer for imaging FAAH in the brain.
ABSTRACT
The application of a phosphoramidate prodrug approach to 2'-C-methylcytidine (NM107), the first nucleoside inhibitor of the hepatitis C virus (HCV) NS5B polymerase, is reported. 2'-C-Methylcytidine, as its valyl ester prodrug (NM283), was efficacious in reducing the viral load in patients infected with HCV. Several of the phosphoramidates prepared demonstrated a 10- to 200-fold superior potency with respect to the parent nucleoside in the cell-based replicon assay. This is due to higher levels of 2'-C-methylcytidine triphosphate in the cells. These prodrugs are efficiently activated and converted to the triphosphate in hepatocytes of several species. Our SAR studies ultimately led to compounds that gave high levels of NTP in hamster and rat liver after subcutaneous dosing and that were devoid of the toxic phenol moiety usually found in ProTides.
Subject(s)
Amides/metabolism , Amides/therapeutic use , Antiviral Agents/metabolism , Cytidine/analogs & derivatives , Hepatitis C/drug therapy , Phosphoric Acids/metabolism , Phosphoric Acids/therapeutic use , Prodrugs/metabolism , Prodrugs/therapeutic use , Amides/pharmacology , Amides/toxicity , Animals , Antiviral Agents/pharmacology , Antiviral Agents/therapeutic use , Antiviral Agents/toxicity , Cell Line , Cytidine/metabolism , Cytidine/pharmacology , Cytidine/therapeutic use , Cytidine/toxicity , Hepacivirus/drug effects , Hepacivirus/physiology , Hepatocytes/drug effects , Hepatocytes/metabolism , Hepatocytes/virology , Humans , Phosphoric Acids/pharmacology , Phosphoric Acids/toxicity , Polyphosphates/metabolism , Prodrugs/pharmacology , Prodrugs/toxicity , Structure-Activity Relationship , Virus Replication/drug effectsABSTRACT
During our effort to design a receptor binding assay to aid in the elucidation of the molecular mechanism of ezetimibe, we prepared a sulfur-35 containing radioligand which exhibits improved potency over the glucuronide conjugate of ezetimibe in both native enterocyte brush border membranes and membranes from cells expressing recombinant NPC1L1. Herein, we describe the different synthetic strategies which were used to obtain this compound as well as its effectiveness in the aforementioned assay.
Subject(s)
Anticholesteremic Agents/chemistry , Azetidines/chemistry , Membrane Proteins/antagonists & inhibitors , Animals , Anticholesteremic Agents/chemical synthesis , Anticholesteremic Agents/pharmacology , Azetidines/chemical synthesis , Azetidines/pharmacology , Cell Line , Ezetimibe , Glucuronides/chemistry , Humans , Ligands , Membrane Proteins/metabolism , Membrane Transport Proteins , Mice , Protein Binding , Rats , Recombinant Proteins/antagonists & inhibitors , Recombinant Proteins/metabolism , Sulfur Radioisotopes/chemistryABSTRACT
The currently approved treatment for hepatitis C virus infections is a combination of Ribavirin and pegylated Interferon. It leads to a sustained virologic response in approximately only half of the patients treated. For this reason there is an urgent need of new therapeutic agents. 2'-C-Methylcytidine is the first nucleoside inhibitor of the HCV NS5B polymerase that was efficacious in reducing the viral load in patients infected with HCV. The application of a monophosphate prodrug approach based on unprecedented cyclic phosphoramidates is reported. Our SAR studies led to compounds that are efficiently converted to the active triphosphate in human hepatocytes.
Subject(s)
Antiviral Agents/metabolism , Antiviral Agents/pharmacology , Cytidine/analogs & derivatives , Hepacivirus/drug effects , Hepatitis C/drug therapy , Prodrugs/pharmacology , Animals , Antiviral Agents/administration & dosage , Antiviral Agents/chemistry , Cricetinae , Cytidine/administration & dosage , Cytidine/chemistry , Cytidine/metabolism , Cytidine/pharmacology , Drug Stability , Hepatocytes/virology , Humans , Prodrugs/administration & dosage , Prodrugs/chemistry , Prodrugs/metabolism , Structure-Activity RelationshipABSTRACT
A very accurate and selective LC-MS/MS method was developed and validated for the quantification of 2'-C-modified nucleoside triphosphate in liver tissue samples. An efficient pretreatment procedure of liver tissue samples was developed, using a fully automated SPE procedure with 96-well SPE plate (weak anion exchange sorbent, 30 mg). Nucleotide hydrophilic interaction chromatography has been performed on an aminopropyl column (100 mm x 2.0 mm, 3 microm) using a gradient mixture of ACN and ACN/water (5:95 v/v) with 20 mM ammonium acetate at pH 9.45 as mobile phase at 300 microL/min flow rate. The 2'-C-modified nucleoside triphosphate was detected in the negative ESI mode in multiple reaction monitoring (MRM) mode. Calibration curve was linear over the 0.05-50 microM concentration range. Satisfying results, confirming the high reliability of the established LC-MS/MS method, were obtained for intraday precision (CV = 2.5-9.1%) and accuracy (92.6-94.8%) and interday precision (CV = 9.6-11.5%) and accuracy (94.4-102.4%) as well as for recovery (82.0-112.6%) and selectivity. The method has been successfully applied for pharmacokinetic studies of 2'-C-methyl-cytidine-triphosphate in liver tissue samples.
Subject(s)
Cytidine Triphosphate/analogs & derivatives , Cytidine Triphosphate/analysis , Liver/chemistry , Tandem Mass Spectrometry/methods , Animals , Antiviral Agents/administration & dosage , Antiviral Agents/metabolism , Calibration , Chromatography, Liquid/methods , Male , Prodrugs/administration & dosage , Prodrugs/metabolism , Pyrimidine Nucleosides/administration & dosage , Pyrimidine Nucleosides/metabolism , Rats , Rats, Sprague-Dawley , Reproducibility of Results , Solid Phase Extraction/methods , Solvents/chemistryABSTRACT
A variety of new prodrugs of 2'-methyl cytidine based on acyloxy ethylamino phosphoramidates have been synthesized and tested in vitro and in vivo for their biological activity. Compared with the parent drug a 10- to 20-fold increase in formation of nucleotide triphosphate in rat and human hepatocytes could be achieved.
Subject(s)
Amides/chemical synthesis , Cytidine/analogs & derivatives , Cytidine/chemical synthesis , Phosphoric Acids/chemical synthesis , Prodrugs/chemical synthesis , RNA-Dependent RNA Polymerase/antagonists & inhibitors , Viral Nonstructural Proteins/antagonists & inhibitors , Amides/pharmacology , Animals , Cells, Cultured , Cytidine/pharmacology , Dogs , Drug Evaluation, Preclinical/methods , Hepacivirus/drug effects , Hepacivirus/enzymology , Hepatocytes/drug effects , Hepatocytes/enzymology , Hepatocytes/virology , Humans , Phosphoric Acids/pharmacology , Prodrugs/pharmacology , RNA-Dependent RNA Polymerase/metabolism , Rabbits , Rats , Viral Nonstructural Proteins/metabolismABSTRACT
As a part of an ongoing medicinal chemistry effort to identify inhibitors of the Hepatitis C Virus RNA replication, we report here the synthesis and biological evaluation of 9-deaza- and 7,9-dideaza-7-oxa-2'-C-methyladenosine. The parent 2'-C-methyladenosine shows excellent intracellular inhibitory activity but poor pharmacokinetic profile. Replacement of the nucleoside-defining 9-N of 2'-C-methyladenosine with a carbon atom was designed to yield metabolically more stable C-nucleosides. Modifications at position 7 were designed to exploit the importance of the hydrogen bond accepting properties of this heteroatom in modulating the adenosine deaminase (ADA) mediated 6-N deamination. 7-Oxa-7,9-dideaza-2'-C-methyladenosine was found to be a moderately active inhibitor of intracellular HCV RNA replication, whereas 9-deaza- 2'-C-methyladenosine showed only weak activity despite excellent overlap of both of the synthesized target compounds with 2'-C-methyladenosine's three dimensional structure. Position 7 of the nucleobase proved to be an effective handle for modulating ADA-mediated degradation, with the rate of degradation correlating with the hydrogen-bonding properties at this position.
Subject(s)
Adenosine/analogs & derivatives , Adenosine/pharmacology , Antiviral Agents/chemistry , Antiviral Agents/pharmacology , Hepacivirus/drug effects , Adenosine/chemistry , Cell Line , Hepacivirus/metabolism , Humans , Models, Molecular , Molecular Structure , RNA, Viral/metabolism , Virus ReplicationABSTRACT
A novel series of oxadiazole based amides have been shown to be potent DPP-4 inhibitors. The optimized compound 43 exhibited excellent selectivity over a variety of DPP-4 homologs.
Subject(s)
Dipeptidyl-Peptidase IV Inhibitors , Oxadiazoles/administration & dosage , Oxadiazoles/pharmacology , Protease Inhibitors/administration & dosage , Protease Inhibitors/pharmacology , Administration, Oral , Animals , Binding Sites , Biological Availability , Dogs , Enzyme Activation/drug effects , Ether-A-Go-Go Potassium Channels/drug effects , Humans , Models, Molecular , Molecular Conformation , Oxadiazoles/chemistry , Protease Inhibitors/chemistry , Protein Conformation , Rats , Stereoisomerism , Structure-Activity RelationshipABSTRACT
A series of beta-substituted biarylphenylalanine amides were synthesized and evaluated as inhibitors of dipeptidyl peptidase IV (DPP-4) for the treatment of type 2 diabetes. Optimization of the metabolic profile of early analogues led to the discovery of (2S,3S)-3-amino-4-(3,3-difluoropyrrolidin-1-yl)-N,N-dimethyl-4-oxo-2-(4-[1,2,4]triazolo[1,5-a]pyridin-6-ylphenyl)butanamide (6), a potent, orally active DPP-4 inhibitor (IC(50) = 6.3 nM) with excellent selectivity, oral bioavailability in preclinical species, and in vivo efficacy in animal models. Compound 6 was selected for further characterization as a potential new treatment for type 2 diabetes.
Subject(s)
Diabetes Mellitus, Type 2/drug therapy , Dipeptidyl Peptidase 4/metabolism , Hypoglycemic Agents/chemical synthesis , Phenylalanine/analogs & derivatives , Protease Inhibitors/chemical synthesis , Triazoles/chemical synthesis , Administration, Oral , Animals , Biological Availability , Calcium Channels, L-Type/drug effects , Cell Line , Crystallography, X-Ray , Glucose Tolerance Test , Humans , Hypoglycemic Agents/chemistry , Hypoglycemic Agents/pharmacology , In Vitro Techniques , Male , Mice , Mice, Inbred C57BL , Mice, Obese , Microsomes, Liver/drug effects , Microsomes, Liver/metabolism , Models, Molecular , Muscle Proteins/antagonists & inhibitors , Muscle, Skeletal/metabolism , NAV1.5 Voltage-Gated Sodium Channel , Phenylalanine/chemical synthesis , Phenylalanine/chemistry , Phenylalanine/pharmacology , Protease Inhibitors/chemistry , Protease Inhibitors/pharmacology , Rabbits , Sodium Channels , Stereoisomerism , Structure-Activity Relationship , Triazoles/chemistry , Triazoles/pharmacologyABSTRACT
anti-Substituted beta-methylphenylalanine derived amides have been shown to be potent DPP-IV inhibitors exhibiting excellent selectivity over both DPP8 and DPP9. The optimized compound exhibited good pharmacokinetic profiles in three preclinical species.
Subject(s)
Adenosine Deaminase Inhibitors , Glycoproteins/antagonists & inhibitors , Protease Inhibitors/administration & dosage , Protease Inhibitors/pharmacology , Pyridones/administration & dosage , Pyridones/pharmacology , Adenosine Deaminase/metabolism , Administration, Oral , Amides/chemistry , Animals , Biological Availability , Dipeptidyl Peptidase 4/metabolism , Dogs , Glycoproteins/metabolism , Humans , Inhibitory Concentration 50 , Macaca mulatta , Molecular Structure , Nitrogen/chemistry , Protease Inhibitors/chemistry , Protease Inhibitors/pharmacokinetics , Pyridones/chemistry , Pyridones/pharmacokinetics , Rats , Sensitivity and Specificity , Structure-Activity Relationship , Substrate SpecificityABSTRACT
anti-Substituted biaryl beta-methylphenylalanine derived amides have been shown to be potent DPP-IV inhibitors that suffer from suboptimal selectivity and pharmacokinetics. This letter describes the substitution of the beta-methyl substituent with beta-polar substituents, culminating in the discovery of a beta-dimethylamide substituted phenylalanine derivative with an excellent potency, selectivity, and pharmacokinetic profile.
