Derivados de cromenopirazoles como ligandos de receptores de cannabinoides

Derivados de cromenopirazoles como ligandos de receptores de cannabinoides. Compuestos derivados de cromenopirazoles que son ligandos de receptores de cannabinoides, su uso para la fabricación de un medicamento, uso de este medicamento para el tratamiento y/o la prevención de trastornos asociados a los receptores de cannabinoides, uso de dicho compuesto como reactivo en ensayos biológicos relacionados con receptores de cannabinoides y procedimiento de obtención de los mismos.Peer reviewedConsejo Superior de Investigaciones Científicas (España), Universidad Complutense de MadridA1 Solicitud de patente con informe sobre el estado de la técnic

Glicosil hidrolasas de Lactiplantibacillus plantarum WCFS1

Resumen del trabajo presentado a la 15ª Reunión de la Red Española de Bacterias Lácticas: Bacterias Lácticas en Alimentación y Salud. Valencia, 26 y 27 de mayo de 2022.AGL2017-84614-C2-1-R y AGL2017-84614-C2-2-RPeer reviewe

Tautomerism of hydroxychromenopyrazoles

Annular tautomerism (OH⋯N and/or NH⋯O) of hydroxychromenopyrazoles was studied by NMR-spectroscopy and B3LYP/6-311++G(d,p) theoretical calculations. The experimental chemical shifts of 4,4-dimethyl-7-pentyl-2,4-dihydrochromeno[4,3-c]pyrazol-9-ol have been compared with absolute shieldings calculations performed using the GIAO approximation. This study shows that the title compound exists mainly as an OH⋯N tautomer in solution. © 2012 Elsevier B.V. All rights reserved.Peer Reviewe

Derivados de cromenopirazoles como ligandos de receptores de cannabinoides.

[EN] The invention relates to compounds derived from cromenopyrazoles that are cannabinoid receptor ligands, to the use thereof for the production of a drug, to the use of this drug in the treatment and/or prevention of disorders associated with cannabinoid receptors, to the use of said compound as a reagent in biological assays related to cannabinoid receptors, and to the method for obtaining same.[ES] Compuestos derivados de cromenopirazoles que son ligandos de receptores de cannabinoides, su uso para la fabricación de un medicamento, uso de este medicamento para el tratamiento y/o la prevención de trastornos asociados a los receptores de cannabinoides, uso de dicho compuesto como reactivo en ensayos biológicos relacionados con receptores de cannabinoides y procedimiento de obtención de los mismos.Peer reviewedConsejo Superior de Investigaciones Científicas (España), Universidad Complutense de MadridA1 Solicitud de patente con informe sobre el estado de la técnic

Chemical modification of novel glycosidases from lactobacillus plantarum using hyaluronic acid: effects on high specificity against 6-Phosphate glucopyranoside

Three novel glycosidases produced from Lactobacillus plantarum, so called Lp_0440, Lp_2777, and Lp_3525, were isolated and overexpressed on Escherichia coli containing a His-tag for specific purification. Their specific activity was evaluated against the hydrolysis of p-nitrophenylglycosides and p-nitrophenyl-6-phosphate glycosides (glucose and galactose) at pH 7. All three were modified with hyaluronic acid (HA) following two strategies: A simple coating by direct incubation at alkaline pH or direct chemical modification at pH 6.8 through preactivation of HA with carbodiimide (EDC) and N-hydroxysuccinimide (NHS) at pH 4.8. The modifications exhibited important effect on enzyme activity and specificity against different glycopyranosides in the three cases. Physical modification showed a radical decrease in specific activity on all glycosidases, without any significant change in enzyme specificity toward monosaccharide (glucose or galactose) or glycoside (C-6 position free or phosphorylated). However, the surface covalent modification of the enzymes showed very interesting results. The glycosidase Lp_0440 showed low glycoside specificity at 25 °C, showing the same activity against p-nitrophenyl-glucopyranoside (pNP-Glu) or p-nitrophenyl-6-phosphate glucopyranoside (pNP-6P-Glu). However, the conjugated cHA-Lp_0440 showed a clear increase in the specificity towards the pNP-Glu and no activity against pNP-6P-Glu. The other two glycosidases (Lp_2777 and Lp_3525) showed high specificity towards pNP-6P-glycosides, especially to the glucose derivative. The HA covalent modification of Lp_3525 (cHA-Lp_3525) generated an enzyme completely specific against the pNP-6P-Glu (phosphoglycosidase) maintaining more than 80% of the activity after chemical modification. When the temperature was increased, an alteration of selectivity was observed. Lp_0440 and cHA-Lp_0440 only showed activity against p-nitrophenyl-galactopyranoside (pNP-Gal) at 40 °C, higher than at 25 °C in the case of the conjugated enzyme.This work was sponsored by the Spanish Government (Project No. AGL2017-84614-C2-2-R) and CSIC Project No. CSIC-PIE 201880E011). We also thank the Ministry of Education, Youth and Sports of the Community of Madrid and the European Social Fund for a contract to C.P.-R (PEJD-2017PRE/SAL-3762) in the program of Youth Employment and the Youth Employment Initiative (YEI) 2017.Peer reviewe

Characterization and improvement of a new glycosidase from Lactobacillus plantarum

Póster presentado a la VII International Conference on Environmental Industrial and Applied Microbiology, celebrada en Madrid (España) del 18 al 20 de octubre de 2017.In recent years, the high stereoselectivity and efficiency of carbohydrate-processing enzymes have been exploited for many biotechnological applications, including flavour enhancement in foods. In recent years, the recent advances in carbohydrate synthesis by glycosidases are based on two complementary approaches: the use of wild-type enzymes with engineered substrates, and mutant glycosidases. In particular, much attention has been focused on the use of β-glucosidases for the enzymatic hydrolysis of flavourless glycoconjugates present in juices and wine beverages for the release aroma volatiles. With the aim to found a novel glycosidase with potential applications in food industry it has been produced and biochemically characterized Lp_3629 glycosidase, a member of GH1 glycosil hydrolase family, from Lactobacillus plantarum. For that purpose, we have clone, using a LIC strategy described previously, and heterologously expressed lp_3629 gene in Escherichia coli. The recombinant Lp_3629 protein containing an amino terminal His6 tag has been produced in a soluble form. Purified recombinant enzyme showed galactosidase but not glucosidase activity. However when 6-phospho-β-D-glucopyranoside was synthetized, it was demonstrated that the enzyme mainly exhibited 6- phospho-β-glucosidase activity. Lp_3629 showed solubility problems since it is a metastable protein. In order to avoid the protein instability, a double mutant Cys211Ser /Cys 292Ser was constructed by site-directed mutagenesis PCR technique. The Lp_3629mutant showed high solubility although it maintain its kinetic parameters.Financial support from the MINEICO (AGL2014-52911-R).Peer Reviewe

Caracterización y mejora de una nueva glicosidasa de la familia GH1 de Lactobacillus plantarum

Póster presentado al IX Congreso CyTA-CESIA, celebrado en Madrid del 16 al 19 de mayo de 2017.Debido a la importancia de las enzimas encargadas de metabalizar carbohidratos en la industria alimentaria, en este trabajo se ha purificado y caracterizado una nueva glicosidasa, Bgl, de L. plantarum CECT748T. Los resultados demuestran que posee actividad 6-P-beta-glucosidasa.Trabajo financiado por el MINEICO (proyecto AGL2014-52911-R).Peer Reviewe

Derivados de 5-NITROINDAZOL y su uso como agentes antiprotozoarios

[EN] The invention relates to two families of 5-nitroindazole derivatives that have antiparasitic properties and to the use thereof for the production of a drug, preferably for the treatment of infections caused by pathogenic protozoa from the Trypanosomatidae and Trichomonadidae families, such as Chagas disease, leishmaniasis and trichomoniasis.[ES] Derivados de 5-NITROINDAZOL y su uso como agentes antiprotozoarios. La presente invención se refiere a dos familias de derivados de 5-NITROINDAZOL que poseen propiedades antiparasitarias y a su empleo para la fabricación de un medicamento, preferentemente para el tratamiento de infecciones causadas por protozoos patógenos de las familias Trypanosomatidae y Trichomonacidae, tales como la enfermedad de Chagas, la leishmaniosis y la tricomonosis.Peer reviewedUniversidad Complutense de Madrid, Consejo Superior de Investigaciones Científicas (España)A1 Solicitud de patente con informe sobre el estado de la técnic

5-NITROINDAZOLE derivatives and use thereof as antiprotozoal agents

[ES] Derivados de 5-NITROINDAZOL y su uso como agentes antiprotozoarios. La presente invención se refiere a dos familias de derivados de 5-NITROINDAZOL que poseen propiedades antiparasitarias y a su empleo para la fabricación de un medicamento, preferentemente para el tratamiento de infecciones causadas por protozoos patógenos de las familias Trypanosomatidae y Trichomonacidae, tales como la enfermedad de Chagas, la leishmaniosis y la tricomonosis.[EN] The invention relates to two families of 5-nitroindazole derivatives that have antiparasitic properties and to the use thereof for the production of a drug, preferably for the treatment of infections caused by pathogenic protozoa from the Trypanosomatidae and Trichomonadidae families, such as Chagas disease, leishmaniasis and trichomoniasis.Peer reviewedUniversidad Complutense de Madrid, Consejo Superior de Investigaciones Científicas (España)B2 Patente con examen previ

Structural basis of the substrate specificity and instability in solution of a glycosidase from Lactobacillus plantarum

Statistics from structural genomics initiatives reveal that around 50–55% of the expressed, non-membrane proteins cannot be purified and therefore structurally characterized due to solubility problems, which emphasized protein solubility as one of the most serious concerns in structural biology projects. Lactobacillus plantarum CECT 748 produces an aggregation-prone glycosidase (LpBgl) that we crystallized previously. However, this result could not be reproduced due to protein instability and therefore further high-resolution structural analyses of LpBgl were impeded. The obtained crystals of LpBgl diffracted up to 2.48 Å resolution and permitted to solve the structure of the enzyme. Analysis of the active site revealed a pocket for phosphate-binding with an uncommon architecture, where a phosphate molecule is tightly bound suggesting the recognition of 6-phosphoryl sugars. In agreement with this observation, we showed that LpBgl exhibited 6-phospho-β-glucosidase activity. Combination of structural and mass spectrometry results revealed the formation of dimethyl arsenic adducts on the solvent exposed cysteine residues Cys211 and Cys292. Remarkably, the double mutant Cys211Ser/Cys292Ser resulted stable in solution at high concentrations indicating that the marginal solubility of LpBgl can be ascribed specifically to these two cysteine residues. The 2.30 Å crystal structure of this double mutant showed no disorder around the newly incorporated serine residues and also loop rearrangements within the phosphate-binding site. Notably, LpBgl could be prepared at high yield by proteolytic digestion of the fusion protein LSLt-LpBgl, which raises important questions about potential hysteretic processes upon its initial production as an enzyme fused to a solubility enhancer.We thank the ESRF (Grenoble, France) for provision of synchrotron radiation facilities (BM16 and ID23-2). Financial support from the Ministerio de Economía y Competitividad (BFU2010-17929/BMC) (to J.M.M.) and AGL2014-52911-R (to R.M.).Peer Reviewe