RESUMEN
Choline kinase (CK) is a homodimeric enzyme that catalyses the transfer of the ATP γ-phosphate to choline, generating phosphocholine and ADP in the presence of magnesium. Several isoforms of CK are present in humans but only the HsCKα has been associated with cancer and validated as a drug target to treat this disease. As a consequence a large number of compounds based on Hemicholinium (HC-3) have been described. Two compounds, previously reported to inhibit the human enzyme, have recently been shown to inhibit P. falciparum CK (PfCK) and therefore their potential applications might be anticipated to other pathogens. Herein, using molecular dynamic simulations, we have firstly observed that the ATP and the choline binding site of different CK in pathogens and human are conserved, suggesting that previous compounds inhibiting the human enzyme may also interact with CKs from different pathogens. We have substantiated such observation with experimental assays showing that HsCKα1, PfCK and CpCK bind to two compounds with distinct structural features in the low µM range. Collectively, these results uncover similarities among the choline kinase binding site from different pathogenic species and the human enzyme, highlighting the feasibility of designing novel inhibitors based on the choline binding pocket.
Asunto(s)
Antiprotozoarios/química , Colina Quinasa/antagonistas & inhibidores , Inhibidores Enzimáticos/química , Hemicolinio 3/análogos & derivados , Proteínas Protozoarias/antagonistas & inhibidores , Adenosina Trifosfato/química , Secuencia de Aminoácidos , Antiprotozoarios/síntesis química , Antiprotozoarios/farmacología , Dominio Catalítico , Colina/química , Colina Quinasa/química , Cryptosporidium parvum/efectos de los fármacos , Cryptosporidium parvum/enzimología , Cryptosporidium parvum/crecimiento & desarrollo , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/farmacología , Hemicolinio 3/síntesis química , Hemicolinio 3/farmacología , Humanos , Concentración 50 Inhibidora , Simulación de Dinámica Molecular , Datos de Secuencia Molecular , Plasmodium falciparum/efectos de los fármacos , Plasmodium falciparum/enzimología , Plasmodium falciparum/crecimiento & desarrollo , Plasmodium knowlesi/efectos de los fármacos , Plasmodium knowlesi/enzimología , Plasmodium knowlesi/crecimiento & desarrollo , Estructura Secundaria de Proteína , Proteínas Protozoarias/química , Homología de Secuencia de Aminoácido , Especificidad de la EspecieRESUMEN
Nitric oxide (NO), which is produced by oxidation of L-arginine to L-citrulline in a process catalyzed by different isoforms of nitric oxide synthase (NOS), exhibits diverse roles in several physiological processes, including neurotransmission, blood pressure regulation and immunological defense mechanisms. On the other hand, an overproduction of NO is related with several disorders as Alzheimer's disease, Huntington's disease and the amyotrophic lateral sclerosis. Taking melatonin as a model, our research group has designed and synthesized several families of compounds that act as NOS inhibitors, and their effects on the excitability of N-methyl-D-aspartate (NMDA)-dependent neurons in rat striatum, and on the activity on both nNOS and iNOS were evaluated. Structural comparison between the three most representative families of compounds (kynurenines, kynurenamines and 4,5-dihydro-1H-pyrazole derivatives) allows the establishment of structure-activity relationships for the inhibition of nNOS, and a pharmacophore model that fulfills all of the observed SARs were developed. This model could serve as a template for the design of other potential nNOS inhibitors. The last family of compounds, pyrrole derivatives, shows moderate in vitro NOS inhibition, but some of these compounds show good iNOS/nNOS selectivity. Two of these compounds, 5-(2-aminophenyl)-1H-pyrrole-2-carboxylic acid methylamide and cyclopentylamide, have been tested as regulators of the in vivo nNOS and iNOS activity. Both compounds prevented the increment of the inducible NOS activity in both cytosol (iNOS) and mitochondria (i-mtNOS) observed in a MPTP model of Parkinson's disease.
Asunto(s)
Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Melatonina/análogos & derivados , Melatonina/farmacología , Óxido Nítrico Sintasa/antagonistas & inhibidores , Animales , Inhibidores Enzimáticos/síntesis química , Humanos , Melatonina/síntesis química , Melatonina/químicaRESUMEN
Farmacia es la Licenciatura que ofrece una mejor formación curricular para aquellos profesionalesque quieren desarrollarse en el campo de la Biomedicina. No existe ninguna otra Licenciatura queaúne conocimientos tan amplios en Ciencias de la Salud, Química, Microbiología, Parasitología,Biología Molecular y Biotecnología, y que aborde la enseñanza de los aspectos relacionados con elmedicamento y las alteraciones fisio/patológicas a un nivel molecular como es la Licenciatura deFarmacia. Para valorar el conocimiento de los alumnos de esta Licenciatura en las diversassalidas profesionales en el área de Biomedicina, hemos realizado un estudio de las necesidades de losmismos basado en encuestas de opinión. Este trabajo ha puesto de manifiesto la necesidad de crear unPlan de Orientación Tutorial (PAT) enfocado a orientarlos sobre salidas profesionales en el área deBiomedicina, de tal manera que se cubran las principales carencias puestas de manifiesto:Motivación: si comprenden la utilidad de la Licenciatura en su futuro profesional se animaránmás a trabajar.Orientación: Fundamentalmente en la elección de asignaturas e itinerarios profesionales relacionadoscon la Biomedicina.Información: Mediante charlas y conferencias en las que profesionales del área de la Biomedicina deentes públicos y privados los orienten sobre las carreras de investigador en ambos ámbitos.Nuestra experiencia como profesores e investigadores nos indica que aquellos alumnos que durantesus estudios de Licenciatura se vinculan a un Departamento como alumnos internos o al menosmuestran algún interés por la investigación, no sólo se orientan profesionalmente mejor sino queaumentan su rendimiento académico significativamente. Este hecho justifica por sí solo la puesta enmarcha del PAT(AU)
The Degree of Pharmacy offers the best instruction for those professionals that want to specialise inthe field of Biomedicine. It doesn't exist any other Degree offering to the students so wide knowledgein Health Sciences, Chemistry, Microbiology, Parasitology, Molecular Biology and Biotechnologythat deals with all the aspects related with drugs and physio/phatological alterations at a molecular level.We have carried out a study of the students requirements based on opinion surveys at the GranadaUniversity. This work has shown the necessity to create a PAT focused on the orientation of thestudents in their professional development in the area of Biomedicine, in such a way that the followingneeds are covered:Motivation: The understanding of the usefulness of the Degree in their professional future willencourage them to work more intensively.Orientation: Mainly in the election of subjects and professional itineraries related with theBiomedicine.Information: Professionals of Biomedicine from both public institutions and private companies willshow them the different aspects of the research in both type institutions by means of chats andconferences.Our experience as professors and researchers shows that those students that are linked to a Departmentduring their Degree or that show some interest for the research, significantly increase both theiracademic and professional performance. This fact justifies the initiation of the PAT(AU)
Asunto(s)
Humanos , Masculino , Femenino , Tutoría/métodos , Tutoría/tendencias , Investigación/organización & administración , Investigación/tendencias , Educación de Postgrado en Farmacia/métodos , Tutoría/organización & administración , Tutoría/normas , Investigación/métodos , Educación de Postgrado en Farmacia/organización & administración , Educación de Postgrado en Farmacia/tendencias , Encuesta SocioeconómicaRESUMEN
This paper describes the unequivocal structural elucidation of a new kind of Delta2-pyrazoline derivatives carried out by means of monodimensional 1H and 13C NMR spectroscopies, bidimensional ones such as HMBC and HMQC experiments, and NOEDIFF effects. Conformational analysis of this molecule agrees very well with the experimentally NOEDIFF effects found.