RESUMEN
A new bioinformatic platform (APTERION) was used to design in a short time and with high specificity an aptamer for the detection of the spike protein, a structural protein of SARS-CoV-2 virus, responsible for the COVID-19 pandemic. The aptamer concentration on the carbon electrode surface was optimized using static contact angle and fluorescence method, while specificity was tested using differential pulse voltammetry (DPV) associated to carbon screen-printed electrodes. The data obtained demonstrated the good features of the aptamer which could be used to create a rapid method for the detection of SARS-CoV-2 virus. In fact, it is specific for spike also when tested against bovine serum albumin and lysozyme, competitor proteins if saliva is used as sample to test for the virus presence. Spectrofluorometric characterization allowed to measure the amount of aptamer present on the carbon electrode surface, while DPV measurements proved the affinity of the aptamer towards the spike protein and gave quantitative results. The acquired data allowed to conclude that the APTERION bioinformatic platform is a good method for aptamer design for rapidity and specificity. KEY POINTS: ⢠Spike protein detection using an electrochemical biosensor ⢠Aptamer characterization by contact angle and fluorescent measurements on electrode surface ⢠Computational design of specific aptamers to speed up the aptameric sequence time.
Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , COVID-19 , Humanos , SARS-CoV-2 , Técnicas Electroquímicas/métodos , Glicoproteína de la Espiga del Coronavirus , Aptámeros de Nucleótidos/química , Pandemias , Carbono , Glicoproteínas , Técnicas Biosensibles/métodos , ElectrodosRESUMEN
A new series of morpholine derivatives has been identified as selective DA D3 receptor antagonists; their in vitro profile and pharmacokinetic data are provided.
Asunto(s)
Antagonistas de los Receptores de Dopamina D2/química , Morfolinas/química , Receptores de Dopamina D3/antagonistas & inhibidores , Animales , Sitios de Unión , Antagonistas de los Receptores de Dopamina D2/síntesis química , Antagonistas de los Receptores de Dopamina D2/farmacocinética , Semivida , Simulación del Acoplamiento Molecular , Morfolinas/síntesis química , Morfolinas/farmacocinética , Estructura Terciaria de Proteína , Ratas , Receptores de Dopamina D3/metabolismoRESUMEN
A novel series of 1,2,4-triazolyl octahydropyrrolo[2,3-b]pyrroles showing high affinity and selectivity at the DA D3 receptor is reported here. Compounds endowed with high selectivity over the hERG channel were identified and their pharmacokinetic properties thoroughly analyzed. A few derivatives with appropriate developability characteristics were selected for further studies and progression along the screening cascade. In particular, derivative 60a, (DA D3 pKi=8.4, DA D2 pKi=6.0 and hERG fpKi=5.2) showed a balanced profile and further refinements are in progress around this molecule.
Asunto(s)
Pirroles/farmacología , Receptores de Dopamina D3/antagonistas & inhibidores , Triazoles/farmacología , Animales , Unión Competitiva/efectos de los fármacos , Células CHO , Cricetulus , Relación Dosis-Respuesta a Droga , Canales de Potasio Éter-A-Go-Go/antagonistas & inhibidores , Células HEK293 , Humanos , Modelos Moleculares , Estructura Molecular , Pirroles/síntesis química , Pirroles/química , Relación Estructura-Actividad , Triazoles/síntesis química , Triazoles/químicaRESUMEN
This article deals with the synthesis and the activities of some 1,5-dialkyl-3-arylureido-1,5-benzodiazepin-2,4-diones which were prepared as potential CCK2 antagonists, with the intention to find a possible follow up of our lead compound GV150013, showing an improved pharmacokinetic profile. The phenyl ring at N-5 was replaced with more hydrophilic substituents, like alkyl groups bearing basic functions. In some cases, the resolution of the racemic key intermediates 3-amino-benzodiazepines was also accomplished. Among the compounds synthesized and characterised so far in this class, the 5-morpholinoethyl derivative 54, was selected as potential follow up of GV150013 and submitted for further evaluation.
Asunto(s)
Benzodiazepinonas/farmacología , Receptor de Colecistoquinina B/antagonistas & inhibidores , Animales , Benzodiazepinonas/síntesis química , Benzodiazepinonas/química , Perros , Cobayas , Ratones , Estructura Molecular , Ratas , Estereoisomerismo , Relación Estructura-Actividad , Distribución TisularRESUMEN
Transient receptor potential cation channel subfamily M member 5 (TRPM5) is a nonselective monovalent cation channel activated by intracellular Ca2+ increase. Within the gastrointestinal system, TRPM5 is expressed in the stoma, small intestine, and colon. In the search for a selective agonist of TRPM5 possessing in vivo gastrointestinal prokinetic activity, a high-throughput screening was performed and compound 1 was identified as a promising hit. Hit validation and hit to lead activities led to the discovery of a series of benzo[d]isothiazole derivatives. Among these, compounds 61 and 64 showed nanomolar activity and excellent selectivity (>100-fold) versus related cation channels. The in vivo drug metabolism and pharmacokinetic profile of compound 64 was found to be ideal for a compound acting locally at the intestinal level, with minimal absorption into systemic circulation. Compound 64 was tested in vivo in a mouse motility assay at 100 mg/kg, and demonstrated increased prokinetic activity.
Asunto(s)
Benzotiazoles/química , Benzotiazoles/farmacología , Tracto Gastrointestinal/metabolismo , Ensayos Analíticos de Alto Rendimiento , Canales Catiónicos TRPM/agonistas , Animales , Benzotiazoles/metabolismo , Benzotiazoles/farmacocinética , Diseño de Fármacos , Tracto Gastrointestinal/efectos de los fármacos , Humanos , Masculino , Ratones , Terapia Molecular Dirigida , Reproducibilidad de los Resultados , Distribución TisularRESUMEN
A novel series of 1,2,4-triazolyl 5-azaspiro[2.4]heptanes with high affinity and selectivity at the dopamine (DA) D3 receptor (D3R) is described. Some of these compounds also have high selectivity over the hERG channel and were characterized with respect to their pharmacokinetic properties both in vitro and in vivo during lead identification and early lead optimization phases. A few derivatives with overall favorable developability characteristics were selected for further late lead optimization studies.
Asunto(s)
Heptanos/química , Heptanos/farmacología , Receptores de Dopamina D3/antagonistas & inhibidores , Compuestos de Espiro/química , Compuestos de Espiro/farmacología , Animales , Células CHO , Cricetulus , Cristalografía por Rayos X , Canales de Potasio Éter-A-Go-Go/metabolismo , Humanos , Modelos Moleculares , Receptores de Dopamina D3/metabolismo , Relación Estructura-Actividad , Triazoles/química , Triazoles/farmacologíaRESUMEN
A series of benzoazepine derivatives, bearing suitable substituents at the C-3 position, was designed and evaluated by superimposition with the pharmacophore model of the glycine binding site. To fully explore the SAR of this class of compounds and to allow the preparation of new different compounds at the C-3 position, appropriate synthetic routes were set up. The benzoazepines were evaluated in terms of in vitro affinity using [3H]glycine binding assay and in vivo potency by inhibition of convulsions induced by N-methyl-D-aspartate (NMDA) in mice. This further analysis confirmed the preliminary results previously reported and that compound 27 is the most promising compound (Ki=32 nM, ED(50)=0.09 mg/kg, i.v.) in this series. Significant neuroprotective effect was observed after both pre- and post-ischaemia administration in the MCAo model. In particular, after post-ischaemia administration, it was found to be still effective when the administration was delayed up to 6 h after occlusion of the middle cerebral artery.
Asunto(s)
Anticonvulsivantes/síntesis química , Azepinas/síntesis química , Fármacos Neuroprotectores/síntesis química , Receptores de Glicina/antagonistas & inhibidores , Receptores de N-Metil-D-Aspartato/antagonistas & inhibidores , Animales , Anticonvulsivantes/química , Anticonvulsivantes/farmacología , Arteriopatías Oclusivas/complicaciones , Azepinas/química , Azepinas/farmacología , Sitios de Unión , Isquemia Encefálica/tratamiento farmacológico , Isquemia Encefálica/etiología , Masculino , Ratones , Arteria Cerebral Media , Modelos Moleculares , Fármacos Neuroprotectores/química , Fármacos Neuroprotectores/farmacología , Ratas , Ratas Sprague-Dawley , Relación Estructura-ActividadRESUMEN
Following the recent disclosure of 3-methyl pyrrole-2,4-dicarboxylic acid 2-propyl ester 4-(1,2,2-trimethyl-propyl) ester, a potent and selective mGluR1 non-competitive antagonist, we report here a detailed exploration of the C-2 position of this scaffold with the preparation of differently substituted amides. Great improvement of the pharmacokinetic properties has been achieved through this exercise.
Asunto(s)
Ésteres/síntesis química , Ésteres/farmacología , Pirroles/síntesis química , Pirroles/farmacología , Receptores de Glutamato Metabotrópico/antagonistas & inhibidores , Amidas/síntesis química , Amidas/farmacocinética , Animales , Azidas/síntesis química , Azidas/farmacocinética , Fenómenos Químicos , Química Física , Cromatografía Líquida de Alta Presión , Cromatografía en Capa Delgada , Ácidos Hidroxámicos/síntesis química , Ácidos Hidroxámicos/farmacocinética , Indicadores y Reactivos , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Ratones , Conformación Molecular , Ratas , Solubilidad , Espectrofotometría Infrarroja , Relación Estructura-ActividadRESUMEN
In an effort to discover novel CRF-1 receptor antagonists exhibiting improved physicochemical properties, a dihydropirrole[2,3]pyridine scaffold was designed and explored in terms of the SAR of the substitution at the pendent phenyl ring and the nature of the heterocyclic moieties present in the upper region of the molecule. Selective and potent compounds have been discovered endowed with reduced ClogP with respect to compounds known in the literature. Of particular relevance was the finding that the in vitro affinity of the series was maintained by reducing the overall lipophilicity. The results achieved by this exploration enabled the formulation of a novel hypothesis on the nature of the receptor binding pocket of this class of CRF-1 receptor antagonists, making use of in silico docking studies of the putative nonpeptidic antagonist binding site set up in house by homology modeling techniques.
Asunto(s)
Simulación por Computador , Piridinas/farmacología , Pirroles/farmacología , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Animales , Sitios de Unión , Células CHO , Cricetinae , Cricetulus , Diseño de Fármacos , Ligandos , Estructura Molecular , Piridinas/síntesis química , Piridinas/química , Pirroles/síntesis química , Pirroles/química , Relación Estructura-Actividad Cuantitativa , EstereoisomerismoRESUMEN
To identify new CRF(1) receptor antagonists, an attempt to modify the bis-heterocycle moiety present in the top region of the dihydropyrrole[2,3]pyridine template was made following new pharmacophoric hypothesis on the CRF(1) receptor antagonists binding pocket. In particular, the 2-thiazole ring, present in the previous series of compounds, was replaced by more hydrophilic non aromatic heterocycles able to make appropriate H-bond interactions with amino acid residues Thr192 and Tyr195. This exploration, followed by an accurate analysis of the substitution of the pendant aryl ring, enabled to identify in vitro potent compounds showing excellent pharmacokinetics and outstanding in vivo activity in animal models of anxiety, both in rodents and primates.
Asunto(s)
Piridinas/síntesis química , Piridinas/farmacología , Pirroles/síntesis química , Pirroles/farmacología , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Animales , Relación Dosis-Respuesta a Droga , Conducta Exploratoria/efectos de los fármacos , Femenino , Miembro Anterior/efectos de los fármacos , Gerbillinae , Humanos , Masculino , Modelos Químicos , Estructura Molecular , Actividad Motora/efectos de los fármacos , Pruebas Psicológicas , Piridinas/química , Pirroles/química , Ratas , Ratas Sprague-Dawley , Estereoisomerismo , Ultrasonido , Vocalización Animal/efectos de los fármacosRESUMEN
A "data mining" methodology based on substructural analysis and standard 1024 Daylight fingerprints as descriptors was applied to a set of known antagonists of a subfamily of ligand-gated ion channels comprising nicotinic acetylcholine receptors (nAChR's), 5-hydroxytryptamine, gamma-amino butyric acid-A, and glycine receptors. The derived scoring function was used to generate a focused set that was screened for alpha7 nAChR, resulting in the identification of novel alpha7 ligands easily amenable to chemical modification. Finally, the same scoring function was applied retrospectively to other in-house sets screened for the same target in the same assay. The results and performance of the method are described in detail.
Asunto(s)
Diseño de Fármacos , Activación del Canal Iónico , Ligandos , Modelos Químicos , Receptores Nicotínicos/química , Algoritmos , Preparaciones Farmacéuticas/químicaRESUMEN
Metabotropic glutamate receptors (mGluRs) are an unusual family of G-protein coupled receptor (GPCR), and are characterised by a large extracellular N-terminal domain that contains the glutamate binding site. We have identified a new class of non-competitive metabotropic glutamate receptor 1 (mGluR1) antagonists, 2,4-dicarboxy-pyrroles which are endowed with nanomolar potency. They interact within the 7 transmembrane (7TM) domain of the receptor and show antinociceptive properties when tested in a number of different animal models.