ABSTRACT
Adaptor protein 2-associated kinase 1 (AAK1) is a member of the Ark1/Prk1 family of serine/threonine kinases and plays a role in modulating receptor endocytosis. AAK1 was identified as a potential therapeutic target for the treatment of neuropathic pain when it was shown that AAK1 knock out (KO) mice had a normal response to the acute pain phase of the mouse formalin model, but a reduced response to the persistent pain phase. Herein we report our early work investigating a series of pyrrolo[2,1-f][1,2,4]triazines as part of our efforts to recapitulate this KO phenotype with a potent, small molecule inhibitor of AAK1. The synthesis, structure-activity relationships (SAR), and in vivo evaluation of these AAK1 inhibitors is described.
ABSTRACT
Asunaprevir (ASV; BMS-650032), a low nanomolar inhibitor of the hepatitis C virus (HCV) NS3 protease, is currently under development, in combination with other direct-acting antiviral (DAA) agents for the treatment of chronic HCV infection. Extensive nonclinical and pharmacokinetic studies have been conducted to characterize the ADME properties of ASV. ASV has a moderate to high clearance in preclinical species. In vitro reaction phenotyping studies demonstrated that the oxidative metabolism of ASV is primarily mediated via CYP3A4; however, studies in bile-duct cannulated rats and dogs suggest that biliary elimination may contribute to overall ASV clearance. ASV is shown to have hepatotropic disposition in all preclinical species tested (liver to plasma ratios >40). The translation of in vitro replicon potency to clinical viral load decline for a previous lead BMS-605339 was leveraged to predict a human dose of 2 mg BID for ASV. Clinical drug-drug interaction (DDI) studies have shown that at therapeutically relevant concentrations of ASV the potential for a DDI is minimal. The need for an interferon free treatment combined with ASV's initial clinical trial data support development of ASV as part of a fixed dose combination for the treatment of patients chronically infected with HCV genotype 1.
Subject(s)
Antiviral Agents/pharmacokinetics , Hepacivirus/enzymology , Isoquinolines/pharmacokinetics , Protease Inhibitors/pharmacokinetics , Sulfonamides/pharmacokinetics , Viral Nonstructural Proteins/antagonists & inhibitors , ATP Binding Cassette Transporter, Subfamily B, Member 1/antagonists & inhibitors , ATP Binding Cassette Transporter, Subfamily B, Member 1/metabolism , Animals , Bile/metabolism , Biological Availability , Cytochrome P-450 CYP3A/metabolism , Cytochrome P-450 Enzyme Inhibitors/pharmacology , Dogs , Hepacivirus/drug effects , Hepatocytes/metabolism , Humans , Liver/metabolism , Macaca fascicularis , Male , Mice , Microsomes, Liver/metabolism , Oxidation-Reduction , Rats , Rats, Sprague-DawleyABSTRACT
7-(2H-Tetrazol-5-yl)-1H-indole 3 was found to be a potent inhibitor of HIV-1 attachment but the compound lacked oral bioavailability in rats. The cause of the low exposure was believed to be poor absorption attributed to the acidic nature of the tetrazole moiety and, in an effort to address this liability, three more lipohilic tetrazole analogs, N-acetoxymethyl 4, N-pivaloyloxymethyl 5, and N-methyl 6, were evaluated as potential oral prodrugs in rats. Prodrug 5 was ineffective in improving the plasma concentration of 3 in vivo but compound 4 provided a 15-fold enhancement of the plasma concentration of 3. Most interestingly, oral dosing of analog 6 afforded a substantial increase in the plasma concentration of the parent in rats when compared to dosing of parent. This represents a novel example of a methyl tetrazole that acts as a prodrug for a free NH tetrazole-containing compound.
Subject(s)
Anti-HIV Agents/chemistry , HIV-1/metabolism , Prodrugs/chemistry , Tetrazoles/chemistry , Administration, Oral , Animals , Anti-HIV Agents/chemical synthesis , Anti-HIV Agents/pharmacokinetics , HIV-1/drug effects , Half-Life , Humans , Prodrugs/chemical synthesis , Prodrugs/pharmacokinetics , Rats , Structure-Activity Relationship , Tetrazoles/chemical synthesis , Tetrazoles/pharmacokinetics , Virus Attachment/drug effectsABSTRACT
As part of the SAR profiling of the indole-oxoacetic piperazinyl benzamide class of HIV-1 attachment inhibitors, substitution at the C7 position of the lead 4-fluoroindole 2 with various 5- and 6-membered heteroaryl moieties was explored. Highly potent (picomolar) inhibitors of pseudotyped HIV-1 in a primary, cell-based assay were identified and select examples were shown to possess nanomolar inhibitory activity against M- and T-tropic viruses in cell culture. These C7-heteroaryl-indole analogs maintained the ligand efficiency (LE) of 2 and were also lipophilic efficient as measured by LLE and LELP. Pharmacokinetic studies of this class of inhibitor in rats showed that several possessed substantially improved IV clearance and half-lives compared to 2. Oral exposure in the rat correlated with membrane permeability as measured in a Caco-2 assay where the highly permeable 1,2,4-oxadiazole analog 13 exhibited the highest exposure.
Subject(s)
Anti-HIV Agents/chemistry , HIV-1/metabolism , Indoles/chemistry , Administration, Oral , Animals , Anti-HIV Agents/chemical synthesis , Anti-HIV Agents/pharmacokinetics , Caco-2 Cells , Cell Membrane Permeability/drug effects , HIV-1/drug effects , Half-Life , Humans , Indoles/chemical synthesis , Indoles/pharmacokinetics , Microsomes, Liver/metabolism , Rats , Structure-Activity Relationship , Virus Attachment/drug effectsABSTRACT
A series of substituted carboxamides at the indole C7 position of the previously described 4-fluoro-substituted indole HIV-1 attachment inhibitor 1 was synthesized and the SAR delineated. Heteroaryl carboxamide inhibitors that exhibited pM potency in the primary cell-based assay against a pseudotype virus expressing a JRFL envelope were identified. The simple methyl amide analog 4 displayed a promising in vitro profile, with its favorable HLM stability and membrane permeability translating into favorable pharmacokinetic properties in preclinical species.
Subject(s)
Amides/chemistry , Anti-HIV Agents/chemistry , HIV-1/metabolism , Indoles/chemistry , Amides/chemical synthesis , Amides/pharmacokinetics , Animals , Anti-HIV Agents/chemical synthesis , Anti-HIV Agents/pharmacokinetics , Biological Availability , Caco-2 Cells , Cell Membrane Permeability/drug effects , Crystallography, X-Ray , Dogs , HIV-1/drug effects , Half-Life , Haplorhini , Humans , Microsomes, Liver/metabolism , Molecular Conformation , Rats , Structure-Activity Relationship , Virus Attachment/drug effectsABSTRACT
Calcitonin gene-related peptide (CGRP) receptor antagonists have demonstrated clinical efficacy in the treatment of acute migraine. Herein, we describe the design, synthesis, and preclinical characterization of a highly potent, oral CGRP receptor antagonist BMS-927711 (8). Compound 8 has good oral bioavailability in rat and cynomolgus monkey, attractive overall preclinical properties, and shows dose-dependent activity in a primate model of CGRP-induced facial blood flow. Compound 8 is presently in phase II clinical trials.
Subject(s)
Calcitonin Gene-Related Peptide/antagonists & inhibitors , Drug Discovery , Migraine Disorders/drug therapy , Piperidines/chemistry , Piperidines/therapeutic use , Pyridines/chemistry , Pyridines/therapeutic use , Administration, Oral , Clinical Trials as Topic , Humans , Magnetic Resonance Spectroscopy , Piperidines/pharmacology , Pyridines/pharmacologyABSTRACT
Calcitonin gene-related peptide (CGRP) receptor antagonists have been clinically shown to be effective in the treatment of migraine, but identification of potent and orally bioavailable compounds has been challenging. Herein, we describe the conceptualization, synthesis, and preclinical characterization of a potent, orally active CGRP receptor antagonist 5 (BMS-846372). Compound 5 has good oral bioavailability in rat, dog, and cynomolgus monkeys and overall attractive preclinical properties including strong (>50% inhibition) exposure-dependent in vivo efficacy in a marmoset migraine model.
ABSTRACT
BACKGROUND: Bioanalytical support of drug-discovery efforts increasingly requires more complex multiple component analysis, including the bioanalysis of drugs, prodrugs and metabolites. Just as the physiochemical properties of these components may differ widely from each other, optimal LC and MS conditions, including polarity, can also vary greatly among the analytes of interest, thus presenting significant challenges during quantitative LC-MS-based bioanalysis. A single compromised method for the determination of all analytes may sacrifice sensitivity or chromatographic conditions for one analyte in order to achieve adequate results for another. Manually switching between assay conditions to analyze samples under separately optimized conditions for individual compounds can be time consuming. RESULTS: The method presented here addresses the problem of differential analyte optimization using a multiplexed approach for simultaneous quantitative bioanalysis of multiple analytes in the same sample, employing a mixed mode of both turbulent- and laminar-flow chromatography. CONCLUSION: The approach is illustrated with the quantitation of a lipophilic drug and its hydrophilic phosphate ester prodrug in a biological matrix under individually optimized LC-MS conditions.
Subject(s)
Chromatography, Liquid/methods , Mass Spectrometry/methods , Phosphates/chemistry , Prodrugs/analysis , Prodrugs/chemistry , Animals , Carbamates/blood , Carbamates/chemistry , Drug Stability , Furans , Hydrophobic and Hydrophilic Interactions , Male , Organophosphates/blood , Organophosphates/chemistry , Rats , Rats, Sprague-Dawley , Sulfonamides/blood , Sulfonamides/chemistry , Time FactorsABSTRACT
A series of novel 5,5-diphenylimidazolones was synthesized and evaluated for activity against the human neuropeptide Y5 receptor. The 3-pyridyl analog 46 demonstrated an IC(50) of 8.3 nM with a favorable pharmacokinetic profile in rats, but was ineffective in reducing food intake.