ABSTRACT
This manuscript describes the discovery of a series of macrocyclic inhibitors of FXIa with oral bioavailability. Assisted by structure based drug design and ligand bound X-ray crystal structures, the group linking the P1 moiety to the macrocyclic core was modified with the goal of reducing H-bond donors to improve pharmacokinetic performance versus 9. This effort resulted in the discovery of several cyclic P1 linkers, exemplified by 10, that are constrained mimics of the bioactive conformation displayed by the acrylamide linker of 9. These cyclic P1 linkers demonstrated enhanced bioavailability and improved potency.
Subject(s)
Drug Design , Drug Discovery , Factor XIa/antagonists & inhibitors , Macrocyclic Compounds/administration & dosage , Macrocyclic Compounds/chemistry , Serine Proteinase Inhibitors/administration & dosage , Serine Proteinase Inhibitors/chemistry , Administration, Oral , Biological Availability , Humans , Ligands , Macrocyclic Compounds/pharmacology , Models, Molecular , Molecular Structure , Serine Proteinase Inhibitors/pharmacology , Structure-Activity RelationshipABSTRACT
We describe a custom Internet of Things (IoT) device used for tracking barcoded containers end to end in a high-throughput analysis and purification laboratory. Our IoT device fills an important gap that previously prevented us from fully tracking barcoded sample containers through manual steps in a multistep workflow, such as when samples are "parked" for temporary storage, or when using instrumentation not otherwise equipped with barcode scanners, a common occurrence found with specific centrifugal evaporation instruments. The custom device reads container barcodes and sends a small amount of data to our back-end data systems. Once data have been received and processed, users are alerted to any system responses via aural and visual feedback. Components of the IoT system include a low-cost headless IoT computer, a barcode reader, and a multicolor LED strip. We believe that the model for our device will facilitate simple and rapid deployment of IoT to the broader laboratory community. All source code and device configurations will be released into the public domain and made freely available.
Subject(s)
Automation, Laboratory/instrumentation , Chemistry Techniques, Analytical/instrumentation , High-Throughput Screening Assays/instrumentation , Internet/instrumentationABSTRACT
Macrocyclic peptides containing N-alkylated amino acids have emerged as a promising therapeutic modality, capable of modulating protein-protein interactions and an intracellular delivery of hydrophilic payloads. While multichannel automated solid-phase peptide synthesis (SPPS) is a practical approach for peptide synthesis, the requirement for slow and inefficient chromatographic purification of the product peptides is a significant limitation to exploring these novel compounds. Herein, we invent a "catch-release" strategy for the nonchromatographic purification of macrocyclic peptides. A traceless catch process is enabled by the invention of a dual-functionalized N-terminal acetate analogue, which serves as a handle for capture onto a purification resin and as a leaving group for macrocyclization. Displacement by a C-terminal nucleophilic side chain thus releases the desired macrocycle from the purification resin. By design, this catch/release process is a logic test for the presence of the key components required for cyclization, thus removing impurities which lack the required functionality, such as common classes of peptide impurities, including hydrolysis fragments and truncated sequences. The method was shown to be highly effective with three libraries of macrocyclic peptides, containing macrocycles of 5-20 amino acids, with either thioether- or amine-based macrocyclic linkages; in this latter class, the reported method represents an enabling technology. In all cases, the catch-release protocol afforded significant enrichment of the target peptides purity, in many cases completely obviating the need for chromatography. Importantly, we have adapted this process for automation on a standard multichannel peptide synthesizer, achieving an efficient and completely integrated synthesis and purification platform for the preparation of these important molecules.
ABSTRACT
A novel series of pyrrolidine-containing GPR40 agonists is described as a potential treatment for type 2 diabetes. The initial pyrrolidine hit was modified by moving the position of the carboxylic acid, a key pharmacophore for GPR40. Addition of a 4-cis-CF3 to the pyrrolidine improves the human GPR40 binding Ki and agonist efficacy. After further optimization, the discovery of a minor enantiomeric impurity with agonist activity led to the finding that enantiomers (R,R)-68 and (S,S)-68 have differential effects on the radioligand used for the binding assay, with (R,R)-68 potentiating the radioligand and (S,S)-68 displacing the radioligand. Compound (R,R)-68 activates both Gq-coupled intracellular Ca2+ flux and Gs-coupled cAMP accumulation. This signaling bias results in a dual mechanism of action for compound (R,R)-68, demonstrating glucose-dependent insulin and GLP-1 secretion in vitro. In vivo, compound (R,R)-68 significantly lowers plasma glucose levels in mice during an oral glucose challenge, encouraging further development of the series.
Subject(s)
Hypoglycemic Agents/pharmacology , Pyrrolidines/pharmacology , Receptors, G-Protein-Coupled/agonists , Animals , Blood Glucose/analysis , Blood Glucose/metabolism , Cell Line , Cells, Cultured , Diabetes Mellitus, Type 2/blood , Diabetes Mellitus, Type 2/drug therapy , Diabetes Mellitus, Type 2/metabolism , Glucagon-Like Peptide 1/metabolism , Humans , Hypoglycemic Agents/chemistry , Hypoglycemic Agents/pharmacokinetics , Hypoglycemic Agents/therapeutic use , Insulin/metabolism , Male , Mice, Inbred C57BL , Models, Molecular , Pyrrolidines/chemistry , Pyrrolidines/pharmacokinetics , Pyrrolidines/therapeutic use , Rats , Receptors, G-Protein-Coupled/metabolismABSTRACT
A new class of Janus kinase (JAK) inhibitors was discovered using a rationally designed pyrrolo[1,2-b]pyridazine-3-carboxamide scaffold. Preliminary studies identified (R)-(2,2-dimethylcyclopentyl)amine as a preferred C4 substituent on the pyrrolopyridazine core (3b). Incorporation of amino group to 3-position of the cyclopentane ring resulted in a series of JAK3 inhibitors (4g-4j) that potently inhibited IFNγ production in an IL2-induced whole blood assay and displayed high functional selectivity for JAK3-JAK1 pathway relative to JAK2. Further modifications led to the discovery of an orally bioavailable (2-fluoro-2-methylcyclopentyl)amino analogue 5g which is a nanomolar inhibitor of both JAK3 and TYK2, functionally selective for the JAK3-JAK1 pathway versus JAK2, and active in a human whole blood assay.
Subject(s)
Drug Discovery , Janus Kinase 1/antagonists & inhibitors , Janus Kinase 2/antagonists & inhibitors , Janus Kinase 3/antagonists & inhibitors , Protein Kinase Inhibitors/chemistry , Protein Kinase Inhibitors/pharmacology , Pyridazines/chemistry , Pyrroles/chemistry , Administration, Oral , Animals , Enzyme-Linked Immunosorbent Assay , Humans , Interferon-gamma/metabolism , Mice , Mice, Inbred BALB C , Models, Molecular , Molecular Structure , Protein Conformation/drug effects , Protein Kinase Inhibitors/administration & dosage , Protein Kinase Inhibitors/pharmacokinetics , Structure-Activity Relationship , Tissue DistributionABSTRACT
A series of diphenylpyridylethanamine-based inhibitors of cholesteryl ester transfer protein with aminoheterocycles appended onto the N-terminus of the chemotype were explored as urea mimetics. Potent compounds were discovered and were further optimized to improve metabolic stability and PXR transactivation profile.
Subject(s)
Cholesterol Ester Transfer Proteins/antagonists & inhibitors , Heterocyclic Compounds/chemical synthesis , Heterocyclic Compounds/pharmacology , Biomimetics , Biphenyl Compounds/chemical synthesis , Biphenyl Compounds/chemistry , Biphenyl Compounds/pharmacology , Drug Stability , Ethylamines/chemical synthesis , Ethylamines/chemistry , Ethylamines/pharmacology , Heterocyclic Compounds/chemistry , Humans , Inhibitory Concentration 50 , Molecular Structure , Pyridines/chemical synthesis , Pyridines/chemistry , Pyridines/pharmacology , Structure-Activity Relationship , Urea/chemistryABSTRACT
Modification of a phenolic lead structure based on lessons learned from increasing the potency of steroidal glucocorticoid agonists lead to the discovery of exceptionally potent, nonsteroidal, indazole GR agonists. SAR was developed to achieve good selectivity against other nuclear hormone receptors with the ultimate goal of achieving a dissociated GR agonist as measured by human in vitro assays. The specific interactions by which this class of compounds inhibits GR was elucidated by solving an X-ray co-crystal structure.
Subject(s)
Amides/chemistry , Amides/pharmacology , Drug Discovery , Receptors, Glucocorticoid/agonists , Binding Sites , Crystallography, X-Ray , Humans , Indazoles/chemistry , Indazoles/pharmacology , Molecular Structure , Protein Binding/drug effects , Steroids/chemistry , Steroids/pharmacology , Structure-Activity RelationshipABSTRACT
A systematic method for assembling and solving complex compound distribution problems is presented in detail. The method is based on a model problem that enumerates the mathematical equations and constraints describing a source container, liquid handler, and three types of destination containers involved in a set of compound distributions. One source container and one liquid handler are permitted in any given problem formulation, although any number of compound distributions may be specified. The relative importance of all distributions is expressed by assigning weights, which are factored into the final mathematical problem specification. A computer program was created that automatically assembles and solves a complete compound distribution problem given the parameters that describe the source container, liquid handler, and any number and type of compound distributions. Business rules are accommodated by adjusting weighting factors assigned to each distribution. An example problem, presented and explored in detail, demonstrates complex and nonintuitive solution behavior.
Subject(s)
Automation, Laboratory/methods , High-Throughput Screening Assays/methods , Specimen Handling/methods , Computer Simulation , SoftwareABSTRACT
A novel series of diphenylpyridylethanamine-based inhibitors of cholesteryl ester transfer protein is described. Optimization of the urea moiety, particularly by incorporation of fluorine, is explored to balance in vitro metabolic stability with CETP potency in the whole plasma assay.
Subject(s)
Cholesterol Ester Transfer Proteins/antagonists & inhibitors , Pyridines/chemistry , Pyridines/pharmacology , Urea/analogs & derivatives , Urea/pharmacology , Amines/blood , Amines/chemistry , Amines/metabolism , Amines/pharmacology , Animals , Cholesterol Ester Transfer Proteins/metabolism , Halogenation , Humans , Mice , Pyridines/blood , Pyridines/metabolism , Rats , Urea/blood , Urea/metabolismABSTRACT
The use of standardized lean manufacturing principles to improve drug discovery productivity is often thought to be at odds with fostering innovation. This manuscript describes how selective implementation of a lean optimized process, in this case centralized purification for medicinal chemistry, can improve operational productivity and increase scientist time available for innovation. A description of the centralized purification process is provided along with both operational and impact (productivity) metrics, which indicate lower cost, higher output, and presumably more free time for innovation as a result of the process changes described.
Subject(s)
Drug Discovery/economics , Pharmaceutical Preparations/economics , Pharmaceutical Preparations/isolation & purification , Chemistry, Pharmaceutical/economics , Chemistry, Pharmaceutical/methods , Chemistry, Pharmaceutical/organization & administration , Drug Discovery/methods , Drug Discovery/organization & administration , Efficiency, Organizational , Humans , Pharmaceutical Preparations/chemistryABSTRACT
A series of diphenylpyridylethanamine (DPPE) derivatives was identified exhibiting potent CETP inhibition. Replacing the labile ester functionality in the initial lead 7 generated a series of amides and ureas. Further optimization of the DPPE series for potency resulted in the discovery of cyclopentylurea 15d, which demonstrated a reduction in cholesterol ester transfer activity (48% of predose level) in hCETP/apoB-100 dual transgenic mice. The PK profile of 15d was suboptimal, and further optimization of the N-terminus resulted in the discovery of amide 20 with an improved PK profile and robust efficacy in transgenic hCETP/apoB-100 mice and in hamsters. Compound 20 demonstrated no significant changes in either mean arterial blood pressure or heart rate in telemeterized rats despite sustained high exposures.
Subject(s)
Anticholesteremic Agents/chemistry , Anticholesteremic Agents/pharmacology , Cholesterol Ester Transfer Proteins/antagonists & inhibitors , Pyridines/chemistry , Pyridines/pharmacology , Stilbenes/chemistry , Stilbenes/pharmacology , Animals , Anticholesteremic Agents/chemical synthesis , Apolipoprotein B-100/antagonists & inhibitors , Apolipoprotein B-100/metabolism , Blood Pressure/drug effects , Cholesterol Ester Transfer Proteins/metabolism , Coronary Disease/drug therapy , Cricetinae , Drug Discovery , Heart Rate/drug effects , Humans , Inhibitory Concentration 50 , Male , Mice , Mice, Transgenic , Molecular Structure , Pyridines/chemical synthesis , Rats , Stilbenes/chemical synthesisABSTRACT
Identification, synthesis and structure-activity relationship of small-molecule VIPR1 antagonists encompassing two chemical series are described.
Subject(s)
Antineoplastic Agents/chemical synthesis , Biphenyl Compounds/chemical synthesis , Receptors, Vasoactive Intestinal Polypeptide, Type I/antagonists & inhibitors , Thiophenes/chemical synthesis , Antineoplastic Agents/pharmacology , Biphenyl Compounds/pharmacology , Cell Line, Tumor , Drug Screening Assays, Antitumor , High-Throughput Screening Assays , Humans , Receptors, Vasoactive Intestinal Polypeptide, Type I/metabolism , Small Molecule Libraries , Structure-Activity Relationship , Thiophenes/pharmacologyABSTRACT
The synthesis, structure-activity relationships (SAR), and biological results of pyridyl-substituted azaindole based tricyclic inhibitors of IKK2 are described. Compound 4m demonstrated potent in vitro potency, acceptable pharmacokinetic and physicochemical properties, and efficacy when dosed orally in a mouse model of inflammatory bowel disease.
Subject(s)
Acetamides/chemistry , Drug Discovery , Enzyme Inhibitors/chemistry , Heterocyclic Compounds, 3-Ring/chemistry , I-kappa B Kinase/antagonists & inhibitors , Acetamides/chemical synthesis , Acetamides/pharmacology , Administration, Oral , Animals , Enzyme Activation/drug effects , Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/pharmacology , Heterocyclic Compounds, 3-Ring/chemical synthesis , Heterocyclic Compounds, 3-Ring/pharmacology , Humans , Inflammatory Bowel Diseases/drug therapy , Inhibitory Concentration 50 , Mice , Molecular Structure , Rats , Structure-Activity RelationshipABSTRACT
SAR studies of pyrrolo[1,2-f]triazines as JAK2 inhibitors is presented. Achieving JAK2 inhibition selectively over JAK3 is discussed.
Subject(s)
Enzyme Activation/drug effects , Enzyme Inhibitors/pharmacology , Janus Kinase 2/metabolism , Janus Kinase 3/metabolism , Pyrrolidines/chemical synthesis , Triazines/chemical synthesis , Triazines/pharmacology , Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/chemistry , Janus Kinase 2/antagonists & inhibitors , Models, Molecular , Molecular Structure , Protein Binding , Pyrroles/chemical synthesis , Pyrroles/chemistry , Pyrroles/pharmacology , Pyrrolidines/chemistry , Pyrrolidines/pharmacology , Structure-Activity Relationship , Triazines/chemistryABSTRACT
The synthesis, structure-activity relationships (SAR) and biological evaluation of thiazole based tricyclic inhibitors of IKK2 are described. Compound 9 was determined to be orally efficacious in a murine model of rheumatoid arthritis.
Subject(s)
Arthritis, Rheumatoid/drug therapy , I-kappa B Kinase/antagonists & inhibitors , Imidazoles/chemistry , Protein Kinase Inhibitors/chemistry , Pyridines/chemistry , Thiazoles/chemistry , Animals , Disease Models, Animal , Dogs , Drug Evaluation, Preclinical , Female , I-kappa B Kinase/metabolism , Mice , Mice, Inbred BALB C , Protein Kinase Inhibitors/pharmacokinetics , Protein Kinase Inhibitors/therapeutic use , Pyridines/pharmacokinetics , Pyridines/therapeutic use , Rats , Structure-Activity RelationshipABSTRACT
Preparative HPLC and HPLC-MS are well established as the methods of choice for purification of pharmaceutical library compounds. Recent advances in supercritical fluid chromatography (SFC) have now made SFC a viable alternative to HPLC for this application. One of the potential arguments for using SFC in place of, or in addition to, HPLC is that it may offer different selectivity and thus has the potential for improved separation success rates. In this paper, we examine relative success rates for SFC and HPLC in obtaining adequate selectivity for successful separation. Our results suggest that use of SFC in addition to HPLC may result in a slight (1-2%) improvement in success rate compared to use of HPLC alone.
Subject(s)
Chromatography, High Pressure Liquid/methods , Chromatography, Supercritical Fluid/methods , Small Molecule Libraries/chemistry , Drug DesignABSTRACT
The design and synthesis of a novel series of oxazole-, thiazole-, and imidazole-based inhibitors of IkappaB kinase (IKK) are reported. Biological activity was improved compared to the pyrazolopurine lead, and the expedient synthesis of the new tricyclic systems allowed for efficient exploration of structure-activity relationships. This, combined with an iterative rat cassette dosing strategy, was used to identify compounds with improved pharmacokinetic (PK) profiles to advance for in vivo evaluation.
Subject(s)
Heterocyclic Compounds, 3-Ring/chemical synthesis , I-kappa B Kinase/antagonists & inhibitors , Imidazoles/chemical synthesis , Oxazoles/chemical synthesis , Thiazoles/chemical synthesis , Animals , Crystallography, X-Ray , Female , Glutathione Transferase/genetics , Glutathione Transferase/metabolism , Heterocyclic Compounds, 3-Ring/pharmacokinetics , Heterocyclic Compounds, 3-Ring/pharmacology , Humans , I-kappa B Kinase/genetics , Imidazoles/pharmacokinetics , Imidazoles/pharmacology , In Vitro Techniques , Leukocytes, Mononuclear/drug effects , Leukocytes, Mononuclear/metabolism , Lipopolysaccharides/pharmacology , Mice , Mice, Inbred BALB C , Microsomes, Liver/metabolism , Oxazoles/pharmacokinetics , Oxazoles/pharmacology , Rats , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Structure-Activity Relationship , Thiazoles/pharmacokinetics , Thiazoles/pharmacology , Tumor Necrosis Factor-alpha/biosynthesisABSTRACT
Several series of pyridine amides were identified as selective and potent 11beta-HSD1 inhibitors. The most potent inhibitors feature 2,6- or 3,5-disubstitution on the pyridine core. Various linkers (CH(2)SO(2), CH(2)S, CH(2)O, S, O, N, bond) between the distal aryl and central pyridyl groups are tolerated, and lipophilic amide groups are generally favored. On the distal aryl group, a number of substitutions are well tolerated. A crystal structure was obtained for a complex between 11beta-HSD1 and the most potent inhibitor in this series.
Subject(s)
11-beta-Hydroxysteroid Dehydrogenase Type 1/antagonists & inhibitors , Amides/chemical synthesis , Amides/pharmacology , Pyridines/chemical synthesis , Pyridines/pharmacology , Amides/chemistry , Combinatorial Chemistry Techniques , Crystallography, X-Ray , Drug Design , Humans , Inhibitory Concentration 50 , Molecular Conformation , Molecular Structure , Pyridines/chemistry , Structure-Activity RelationshipABSTRACT
2-Arylbenzoxazole 5 was identified as a hit from a fluorescence-based high-throughput screen for CETP inhibitors. The synthesis and SAR investigation employing array synthesis of the A- and B-rings are described.