RESUMEN
Systemic fungal infections represent an important public health concern, and new antifungal agents are highly desirable. Herein, we describe the design, synthesis, and biological evaluation of a novel class of antifungal compounds called antibody-recruiting molecules targeting fungi (ARM-Fs). Our approach relies on the use of non-peptidic small molecules, which selectively bind fungal cells and recruit endogenous antibodies to their surfaces, resulting in immune-mediated clearance. Using the opportunistic fungal pathogen Candida albicans as a model, we identified a highly specific bifunctional molecule able to mediate the engulfment and phagocytosis of C. albicans cells by human immune cells in biologically relevant functional assays. This work represents a novel therapeutic approach to treating fungal illness with significant potential to complement and/or combine with existing treatment strategies.
Asunto(s)
Anticuerpos/inmunología , Subgrupos de Linfocitos B/inmunología , Candida albicans/inmunología , Acetilglucosamina/química , Anticuerpos/metabolismo , Subgrupos de Linfocitos B/citología , Candida albicans/metabolismo , Candida albicans/patogenicidad , Quitina/química , Quitina/metabolismo , Dinitrobencenos/química , Dinitrobencenos/inmunología , Diseño de Fármacos , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Células HL-60 , Interacciones Huésped-Patógeno , Humanos , FagocitosisRESUMEN
The reactivity of a representative set of 17 organozinc pivalates with 18 polyfunctional druglike electrophiles (informers) in Negishi cross-coupling reactions was evaluated by high-throughput experimentation protocols. The high-fidelity scaleup of successful reactions in parallel enabled the isolation of sufficient material for biological testing, thus demonstrating the high value of these new solid zinc reagents in a drug-discovery setting and potentially for many other applications in chemistry. Principal component analysis (PCA) clearly defined the independent roles of the zincates and the informers toward druggable-space coverage.
Asunto(s)
Compuestos Organometálicos/química , Piridinas/síntesis química , Zinc/química , Ensayos Analíticos de Alto Rendimiento , Estructura Molecular , Análisis de Componente Principal , Piridinas/químicaRESUMEN
This paper brings together the concepts of molecular complexity and crowdsourcing. An exercise was done at Merck where 386 chemists voted on the molecular complexity (on a scale of 1-5) of 2681 molecules taken from various sources: public, licensed, and in-house. The meanComplexity of a molecule is the average over all votes for that molecule. As long as enough votes are cast per molecule, we find meanComplexity is quite easy to model with QSAR methods using only a handful of physical descriptors (e.g., number of chiral centers, number of unique topological torsions, a Wiener index, etc.). The high level of self-consistency of the model (cross-validated R(2) â¼0.88) is remarkable given that our chemists do not agree with each other strongly about the complexity of any given molecule. Thus, the power of crowdsourcing is clearly demonstrated in this case. The meanComplexity appears to be correlated with at least one metric of synthetic complexity from the literature derived in a different way and is correlated with values of process mass intensity (PMI) from the literature and from in-house studies. Complexity can be used to differentiate between in-house programs and to follow a program over time.
Asunto(s)
Colaboración de las Masas , Estructura Molecular , Bases de Datos de Compuestos Químicos , Humanos , Modelos Químicos , Relación Estructura-Actividad Cuantitativa , EstereoisomerismoRESUMEN
The EphA4 receptor and its ephrin ligands are involved in astrocytic gliosis following CNS injury. Therefore, a strategy aimed at the blockade of EphA4 signaling could have broad therapeutic interest in brain disorders. We have identified novel small molecule inhibitors of EphA4 kinase in specific enzymatic and cell-based assays. In addition, we have demonstrated in two in vitro models of scratch injury that EphA4 receptor kinase is activated through phosphorylation and is involved in the repopulation of the wound after the scratch. A potent EphA4 kinase inhibitor significantly inhibited wound closure and reduced the accumulation of the reactive astrocytes inside the scratch. We have also shown that after the transient focal cerebral ischemia in rats, a large glial scar is formed by the accumulation of astrocytes and chondroitin sulfate proteoglycan surrounding the infarcted tissue at 7 days and 14 days of reperfusion. EphA4 protein expression is highly up-regulated in the same areas at these time points, supporting its potential role in the glial scar formation and maintenance. Taken together, these results suggest that EphA4 kinase inhibitors might interfere with the astrogliosis reaction and thereby lead to improved neurological outcome after ischemic injury.
Asunto(s)
Gliosis/tratamiento farmacológico , Inhibidores de Proteínas Quinasas/uso terapéutico , Receptor EphA4/antagonistas & inhibidores , Heridas y Lesiones/patología , Animales , Astrocitos/patología , Western Blotting , Células CHO , Movimiento Celular/efectos de los fármacos , Células Cultivadas , Cricetinae , Cricetulus , Gliosis/patología , Humanos , Inmunohistoquímica , Ataque Isquémico Transitorio/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Microscopía Fluorescente , Ratas , Ratas Sprague-Dawley , Bibliotecas de Moléculas Pequeñas , Cicatrización de Heridas/efectos de los fármacosRESUMEN
This Letter describes the one pot synthesis of tertiary carbinamine 3 and related analogs of brain penetrant BACE-1 inhibitors via the alkylation of the Schiff base intermediate 2. The methodology developed for this study provided a convenient and rapid means to explore the P1 region of these types of inhibitors, where the P1 group is installed in the final step using a one-pot two-step protocol. Further SAR studies led to the identification of 10 which is twofold more potent in vitro as compared to the lead compound. This inhibitor was characterized in a cisterna magna ported rhesus monkey model, where significant lowering of CSF Abeta40 was observed.
Asunto(s)
Secretasas de la Proteína Precursora del Amiloide/antagonistas & inhibidores , Ácido Aspártico Endopeptidasas/antagonistas & inhibidores , Encéfalo/enzimología , Inhibidores Enzimáticos/química , Oxadiazoles/química , Sulfonamidas/química , Secretasas de la Proteína Precursora del Amiloide/metabolismo , Péptidos beta-Amiloides/metabolismo , Animales , Ácido Aspártico Endopeptidasas/metabolismo , Encéfalo/metabolismo , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/farmacocinética , Humanos , Macaca mulatta , Oxadiazoles/síntesis química , Oxadiazoles/farmacocinética , Fragmentos de Péptidos/metabolismo , Relación Estructura-Actividad , Sulfonamidas/síntesis química , Sulfonamidas/farmacocinéticaRESUMEN
The optimization of tertiary carbinamine derived inhibitors of BACE1 from its discovery as an unstable lead to low nanomolar cell active compounds is described. Five-membered heterocycles are reported as stable and potency enhancing linkers. In the course of this work, we have discovered a clear trend where the activity of inhibitors at a given assay pH is dependent on pK(a) of the amino group that interacts directly with the catalytic aspartates. The potency of compounds as inhibitors of Alphabeta production in a cell culture assay correlated much better with BACE1 enzyme potency measured at pH 7.5 than at pH 4.5.
Asunto(s)
Aminas/metabolismo , Secretasas de la Proteína Precursora del Amiloide/antagonistas & inhibidores , Ácido Aspártico Endopeptidasas/antagonistas & inhibidores , Ácido Aspártico/metabolismo , Inhibidores Enzimáticos/farmacología , Catálisis , Humanos , Modelos Moleculares , Relación Estructura-ActividadRESUMEN
We describe the discovery and optimization of tertiary carbinamine derived inhibitors of the enzyme beta-secretase (BACE-1). These novel non-transition-state-derived ligands incorporate a single primary amine to interact with the catalytic aspartates of the target enzyme. Optimization of this series provided inhibitors with intrinsic and functional potency comparable to evolved transition state isostere derived inhibitors of BACE-1.
Asunto(s)
Secretasas de la Proteína Precursora del Amiloide/antagonistas & inhibidores , Secretasas de la Proteína Precursora del Amiloide/química , Compuestos de Anilina/síntesis química , Oxadiazoles/síntesis química , Compuestos de Anilina/química , Cristalografía por Rayos X , Modelos Moleculares , Oxadiazoles/químicaRESUMEN
A novel HIV protease inhibitor was designed using a morpholine core as the aspartate binding group. Analysis of the crystal structure of the initial lead bound to HIV protease enabled optimization of enzyme potency and antiviral activity. This afforded a series of potent orally bioavailable inhibitors of which MK-8718 was identified as a compound with a favorable overall profile.
RESUMEN
Optimization of a previously reported thrombin inhibitor, 9-hydroxy-9-fluorenylcarbonyl-l-prolyl-trans-4-aminocyclohexylmethylamide (1), by replacing the aminocyclohexyl P1 group provided a new lead structure, 9-hydroxy-9-fluorenylcarbonyl-l-prolyl-2-aminomethyl-5-chlorobenzylamide (2), with improved potency (K(i) = 0.49 nM for human thrombin, 2x APTT = 0.37 microM in human plasma) and pharmacokinetic properties (F = 39%, iv T(1/2) = 13 h in dogs). An effective strategy for reducing plasma protein binding of 2 and improving efficacy in an in vivo thrombosis model in rats was to replace the lipophilic fluorenyl group in P3 with an azafluorenyl group. Systematic investigation of all possible azafluorenyl P3 isomers and azafluorenyl-N-oxide analogues of 2 led to the identification of an optimal compound, 3-aza-9-hydroxyfluoren-9(R)-ylcarbonyl-l-prolyl-2-aminomethyl-5-chlorobenzylamide (19b), with high potency (K(i) = 0.40 nM, 2x APTT = 0.18 microM), excellent pharmacokinetic properties (F = 55%, T(1/2) = 14 h in dogs), and complete efficacy in the in vivo thrombosis model in rats (inhibition of FeCl(3)-induced vessel occlusions in six of six rats receiving an intravenous infusion of 10 microg/kg/min of 19b). The stereochemistry of the azafluorenyl group in 19b was determined by X-ray crystallographic analysis of its N-oxide derivative (23b) bound in the active site of human thrombin.
Asunto(s)
Fluorenos/síntesis química , Prolina/análogos & derivados , Prolina/síntesis química , Trombina/antagonistas & inhibidores , Administración Oral , Animales , Disponibilidad Biológica , Proteínas Sanguíneas/metabolismo , Cristalografía por Rayos X , Perros , Fluorenos/química , Fluorenos/farmacología , Semivida , Humanos , Técnicas In Vitro , Macaca mulatta , Masculino , Microsomas Hepáticos/metabolismo , Modelos Moleculares , Prolina/química , Prolina/farmacología , Unión Proteica , Ratas , Ratas Sprague-Dawley , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
Thrombin-activatable fibrinolysis inhibitor (TAFI) is an important regulator of fibrinolysis, and inhibitors of this enzyme have potential use in antithrombotic and thrombolytic therapy. Appropriately substituted imidazole acetic acids such as 10j were found to be potent inhibitors of activated TAFI and selective versus the related carboxypeptidases CPA, CPN, and CPM but not CPB. Further, 10j accelerated clot lysis in vitro and was shown to be efficacious in a primate model of thrombosis.
Asunto(s)
Acetatos/síntesis química , Aminopiridinas/síntesis química , Carboxipeptidasa B2/antagonistas & inhibidores , Fibrinolíticos/síntesis química , Imidazoles/síntesis química , Propionatos/síntesis química , Inhibidores de Proteasas/síntesis química , Acetatos/farmacocinética , Acetatos/farmacología , Aminopiridinas/farmacocinética , Aminopiridinas/farmacología , Animales , Sitios de Unión , Carboxipeptidasa B2/química , Perros , Fibrinolíticos/farmacocinética , Fibrinolíticos/farmacología , Humanos , Imidazoles/farmacocinética , Imidazoles/farmacología , Técnicas In Vitro , Microsomas/metabolismo , Modelos Moleculares , Propionatos/farmacocinética , Propionatos/farmacología , Inhibidores de Proteasas/farmacocinética , Inhibidores de Proteasas/farmacología , Ratas , Relación Estructura-ActividadRESUMEN
In an effort to discover potent, clinically useful thrombin inhibitors, a rapid analogue synthetic approach was used to explore the P(1) region. Various benzylamines were coupled to a pyridine/pyrazinone P(2)-P(3) template. One compound with an o-thiadiazole benzylic substitution was found to have a thrombin K(i) of 0.84 nM. A study of ortho-substituted five-membered-ring heterocycles was undertaken and subsequently demonstrated that the o-triazole and tetrazole rings were optimal. Combination of these potent P(1) aryl heterocycles with a variety of P(2)-P(3) groups produced a compound with an extraordinary thrombin inhibitory activity of 1.4 pM. It is hoped that this potency enhancement in P(1) will allow for more diversification in the P(2)-P(3) region to ultimately address additional pharmacological concerns.
Asunto(s)
Compuestos Heterocíclicos/síntesis química , Trombina/antagonistas & inhibidores , Bencilaminas/síntesis química , Bencilaminas/química , Sitios de Unión , Compuestos Heterocíclicos/química , Modelos Moleculares , Pirazinas/síntesis química , Pirazinas/química , Piridinas/síntesis química , Piridinas/química , Relación Estructura-Actividad , Tetrazoles/síntesis química , Tetrazoles/química , Tiadiazoles/síntesis química , Tiadiazoles/química , Trombina/química , Triazoles/síntesis química , Triazoles/químicaRESUMEN
This letter describes replacements for the P3 amide moiety present in previously reported tertiary carbinamine macrolactones. Although P-gp efflux issues associated with these amide-macrolactones were solved and full brain penetration was measured in one case, potency was compromised in the process.
Asunto(s)
Aminas/farmacocinética , Secretasas de la Proteína Precursora del Amiloide/antagonistas & inhibidores , Ácido Aspártico Endopeptidasas/antagonistas & inhibidores , Encéfalo/metabolismo , Inhibidores Enzimáticos/farmacocinética , Cristalografía por Rayos X , Modelos MolecularesRESUMEN
BACE-1 is a flexible enzyme with experimentally determined motion in the flap region, the catalytic aspartates, and the 10s loop. Four in-house crystallographically determined complexes of tertiary carbinamine inhibitors revealed 10s loop motion in the S(3) pocket. These X-ray structures were used to correlate K(i) values, which span over five orders of magnitude, with the calculated interaction energy, using the Merck Molecular Force Field for a series of 19 tertiary carbinamine inhibitors.
Asunto(s)
Secretasas de la Proteína Precursora del Amiloide/antagonistas & inhibidores , Inhibidores Enzimáticos/farmacología , Ácido Aspártico/análogos & derivados , Ácido Aspártico/química , Ácido Aspártico/farmacología , Sitios de Unión , Catálisis , Fenómenos Químicos , Química Física , Cristalografía por Rayos X , Inhibidores Enzimáticos/química , Ligandos , Conformación Molecular , Relación Estructura-Actividad , TermodinámicaRESUMEN
A series of low-molecular weight 2,6-diamino-isonicotinamide BACE-1 inhibitors containing an amine transition-state isostere were synthesized and shown to be highly potent in both enzymatic and cell-based assays. These inhibitors contain a trans-S,S-methyl cyclopropane P(3) which bind BACE-1 in a 10s-loop down conformation giving rise to highly potent compounds with favorable molecular weight and moderate to high susceptibility to P-glycoprotein (P-gp) efflux.
Asunto(s)
Secretasas de la Proteína Precursora del Amiloide/antagonistas & inhibidores , Ácido Aspártico Endopeptidasas/antagonistas & inhibidores , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/farmacología , Niacinamida/síntesis química , Niacinamida/farmacología , Animales , Baculoviridae/efectos de los fármacos , Baculoviridae/enzimología , Disponibilidad Biológica , Células Cultivadas , Espectroscopía de Resonancia Magnética , Modelos Moleculares , Conformación Molecular , Peso Molecular , Niacinamida/farmacocinética , Ratas , Relación Estructura-ActividadRESUMEN
This Letter describes the design and synthesis of tertiary carbinamine macrocyclic inhibitors of the beta-secretase (BACE-1) enzyme. These macrocyclic inhibitors, some of which incorporate novel P2 substituents, display a 2- to 100-fold increase in potency relative to the previously described acyclic analogs while affording greater stability.
Asunto(s)
Aminas/química , Aminas/farmacología , Secretasas de la Proteína Precursora del Amiloide/antagonistas & inhibidores , Ácido Aspártico Endopeptidasas/antagonistas & inhibidores , Inhibidores de Proteasas/química , Inhibidores de Proteasas/farmacología , Aminas/síntesis química , Ciclización , Diseño de Fármacos , Modelos Moleculares , Inhibidores de Proteasas/síntesis química , Relación Estructura-ActividadRESUMEN
In this study, we have demonstrated that the critical hydrogen bonding motif of the established 3-aminopyrazinone thrombin inhibitors can be effectively mimicked by a 2-aminopyridine N-oxide. As this peptidomimetic core is more resistant toward oxidative metabolism, it also overcomes the metabolic liability associated with the pyrazinones. An optimization study of the P(1) benzylamide delivered the potent thrombin inhibitor 21 (K(i) = 3.2 nM, 2xaPTT = 360 nM), which exhibited good plasma levels and half-life after oral dosing in the dog (C(max) = 2.6 microM, t(1/2) = 4.5 h).
Asunto(s)
Antitrombinas/química , Pirimidinas/química , Enlace de Hidrógeno , Modelos Moleculares , Imitación MolecularRESUMEN
Structural modifications of the aminopyridine P(1)(') group of imidazole acetic acid based TAFIa inhibitors led to the discovery of the aminocyclopentyl analog 28, a 1 nM TAFIa inhibitor with CLT(50) functional activity of 14 nM but without selectivity against CPB. While not as active, aminobutyl derivative 27 provided an improved 6.7-fold selectivity for TAFIa versus CPB.
Asunto(s)
Carboxipeptidasa B2/antagonistas & inhibidores , Inhibidores Enzimáticos/farmacología , Imidazoles/farmacología , Inhibidores Enzimáticos/química , Imidazoles/química , Modelos Moleculares , Relación Estructura-ActividadRESUMEN
The synthesis and biological evaluation of a series of nonpeptidic small molecule antagonists of the human platelet thrombin receptor (PAR-1) are described. Optimization of the 5-amino-3-arylisoxazole lead resulted in an approximate 100-fold increase in potency. The most potent of these compounds (54) inhibits platelet activation with IC(50)s of 90 nM against the thrombin receptor agonist peptide (TRAP) and 510 nM against thrombin as the agonist. Further, antagonist 54 fully blocks platelet aggregation stimulated by 1 nM thrombin for 10 min.
Asunto(s)
Azepinas/síntesis química , Azepinas/farmacología , Plaquetas/metabolismo , Isoxazoles/síntesis química , Isoxazoles/farmacología , Receptores de Trombina/antagonistas & inhibidores , Plaquetas/efectos de los fármacos , Diseño de Fármacos , Humanos , Técnicas In Vitro , Indicadores y Reactivos , Fragmentos de Péptidos/antagonistas & inhibidores , Fragmentos de Péptidos/farmacología , Piperidinas/síntesis química , Piperidinas/química , Activación Plaquetaria/efectos de los fármacos , Agregación Plaquetaria/efectos de los fármacos , Inhibidores de Agregación Plaquetaria/síntesis química , Inhibidores de Agregación Plaquetaria/farmacología , Receptor PAR-1 , Relación Estructura-Actividad , Trombina/antagonistas & inhibidoresRESUMEN
A series of potent and selective proline- and pyrazinone-based macrocyclic thrombin inhibitors is described. Detailed SAR studies led to the incorporation of specific functional groups in the tether that enhanced functional activity against thrombin and provided exquisite selectivity against trypsin and tPA. X-ray crystallography and molecular modeling studies revealed the inhibitor-enzyme interactions responsible for this selectivity.