Tetrahydropyrazolo[1,5-a]Pyrimidine-3-Carboxamide and N-Benzyl-6′,7′-Dihydrospiro[Piperidine-4,4′-Thieno[3,2-c]Pyran] analogues with bactericidal efficacy against Mycobacterium tuberculosis targeting MmpL3

Mycobacterium tuberculosis is a major human pathogen and the causative agent for the pulmonary disease, tuberculosis (TB). Current treatment programs to combat TB are under threat due to the emergence of multi-drug and extensively-drug resistant TB. As part of our efforts towards the discovery of new anti-tubercular leads, a number of potent tetrahydropyrazolo[1,5-a]pyrimidine-3-ca​rboxamide(THPP) and N-benzyl-6′,7′-dihydrospiro[piperidine-4,​4′-thieno[3,2-c]pyran](Spiro) analogues were recently identified against Mycobacterium tuberculosis and Mycobacterium bovis BCG through a high-throughput whole-cell screening campaign. Herein, we describe the attractive in vitro and in vivo anti-tubercular profiles of both lead series. The generation of M. tuberculosis spontaneous mutants and subsequent whole genome sequencing of several resistant mutants identified single mutations in the essential mmpL3 gene. This ‘genetic phenotype’ was further confirmed by a ‘chemical phenotype’, whereby M. bovis BCG treated with both the THPP and Spiro series resulted in the accumulation of trehalose monomycolate. In vivo efficacy evaluation of two optimized THPP and Spiro leads showed how the compounds were able to reduce >2 logs bacterial cfu counts in the lungs of infected mice

Easy-To-Synthesize Spirocyclic Compounds Possess Remarkable in Vivo Activity against Mycobacterium tuberculosis

Society urgently needs new, effective medicines for the treatment of tuberculosis. To kick-start the required hit-to-lead campaigns, the libraries of pharmaceutical companies have recently been evaluated for starting points. The GlaxoSmithKline (GSK) library yielded many high-quality hits, and the associated data were placed in the public domain to stimulate engagement by the wider community. One such series, the spiro compounds, are described here. The compounds were explored by a combination of traditional in-house research and open source methods. The series benefits from a particularly simple structure and a short associated synthetic chemistry route. Many members of the series displayed striking potency and low toxicity, and highly promising in vivo activity in a mouse model was confirmed with one of the analogues. Ultimately the series was discontinued due to concerns over safety, but the associated data remain public domain, empowering others to resume the series if the perceived deficiencies can be overcome

A GIS-supported Multidisciplinary Database for the Management of UNESCO Global Geoparks: the Courel Mountains Geopark (Spain)

[Abstract] The management of a UNESCO Global Geopark (UGGp) requires a vast wealth of miscellaneous scientific knowledge that can be successfully organised using a Geographical Information System (GIS). This paper presents a pragmatic GIS database to assist in the suitable governance of the Courel Mountains UGGp (2017) in Northwest Spain. The database is structured in 66 coverages compiled from public sources and previous works or produced through traditional mapping (combining fieldwork and photointerpretation) and GIS tools. The acquired data was later homogenised and validated by a multidisciplinary team and archived in independent coverages. Forty thematic maps illustrate the broad range of cartographic information included in the GIS database. Among them, 25 basic maps provide an overview of the UGGp and 15 new maps focus on crosscutting and technical issues. All maps illustrate the huge potential of GIS to create new resources combining coverages and adapting the legend according to their purpose and audience. The database facilitates the suitable publishing of consistent outputs (e.g., brochures, books, panels, webpages, web serves), as well as the elaboration of technical data to assist the park management. The database furnishes information on the design of education actions, touristic routes, activities and Geopark facilities. The GIS database is also a supportive tool for scientific research and provides the necessary knowledge to conduct geoconservation actions based on land use, geological hazards and the occurrence of natural and cultural heritages. Altogether, the GIS database constitutes a powerful instrument for policy-making, facilitating the identification and evaluation of alternative strategy plans.This work was developed in the framework of the Scientific Program of the Courel Mountains UGGp with the cooperation of tourism agents (A.M. Arza and A. López), roofing slate quarries (Pizarras de Villarbacú, Pizarras de Quiroga) and local people (M. Reinosa, G. Díaz, O. Álvarez). We are deeply grateful to J.R. Martínez Catalán (Universidad de Salamanca), A. Pérez-Alberti and J. Guitián (both from Universidade de Santiago de Compostela), J.R. Gutiérrez-Marco (ICOG, Universidad Complutense de Madrid/CSIC), J. Vegas (IGME-CSIC), L. González-Menéndez (IGME-CSIC), J.M. García Queijeiro (Universidade de Vigo), L. Santos and A. Grandal-D’Anglade (both from Universidade da Coruña) for their assistance supplying information involved in the database. We thank also E. de Boer for proofreading the article. DB is grant holder of Plan Andaluz de Investigación, Desarrollo e Innovación 2021, funded by Junta de Andalucí

The Courel Mountains UNESCO Global Geopark: An Amazing Geological History Extended Along 600 Million Years

[EN] The Courel Mountains UNESCO Global Geopark (2019) stands out in SW of Europe because of its geoheritage, its biodiversity and its cultural heritage, all of it considered of international interest. These aspects shape the local development economic and cultural improvement and development. The geoheritage is the result of three geological cycles since the Proterozoic, involving the Cadomian-Avalonian-Pan-African orogeny, the opening of the Rheic Ocean and the Variscan orogeny, and finally the Permian-Mesozoic continental expansion and the Alpine orogeny. The geological history of Courel Mountains is one of singular rocks, huge recumbent folds, valuable metallic mineralization, and invertebrate fossils preserved within metamorphic rocks. This long history is recorded in an exceptional Variscan basement that we can ravel thanks to the exhumation during the Alpine uplifting, when the present-day Courel Mountains were built.Peer reviewe

Biochemical and structural characterization of mycobacterial aspartyl-tRNA synthetase AspS, a promising TB drug target.

The human pathogen Mycobacterium tuberculosis is the causative agent of pulmonary tuberculosis (TB), a disease with high worldwide mortality rates. Current treatment programs are under significant threat from multi-drug and extensively-drug resistant strains of M. tuberculosis, and it is essential to identify new inhibitors and their targets. We generated spontaneous resistant mutants in Mycobacterium bovis BCG in the presence of 10× the minimum inhibitory concentration (MIC) of compound 1, a previously identified potent inhibitor of mycobacterial growth in culture. Whole genome sequencing of two resistant mutants revealed in one case a single nucleotide polymorphism in the gene aspS at 535GAC>535AAC (D179N), while in the second mutant a single nucleotide polymorphism was identified upstream of the aspS promoter region. We probed whole cell target engagement by overexpressing either M. bovis BCG aspS or Mycobacterium smegmatis aspS, which resulted in a ten-fold and greater than ten-fold increase, respectively, of the MIC against compound 1. To analyse the impact of inhibitor 1 on M. tuberculosis AspS (Mt-AspS) activity we over-expressed, purified and characterised the kinetics of this enzyme using a robust tRNA-independent assay adapted to a high-throughput screening format. Finally, to aid hit-to-lead optimization, the crystal structure of apo M. smegmatis AspS was determined to a resolution of 2.4 Å

Preliminary Hydrogeological Investigations for Sustainable Development in the Courel Mountains UNESCO Global Geopark (NW Spain)

Congreso realizado en Málaga en junio de 2022.[EN] Karst aquifers and, especially, their caves and springs, are singular sites of the geoheritage due to their scientific, touristic and use (water resources) values. Besides, they are recognized as habitats of special protection by the Habitat Directive (92/43/CEE) and would be used for sustainable development in UNESCO Global Geoparks (UGGp). However, their management continues to be a challenge at the present day since karst aquifers and their associated springs and caves are vulnerable environments used by many actors and affected by land uses and changes in surface hydrology and groundwater. Our work aims to implement of a suitable management of aquifers and springs in the Courel Mountains UGGp based on hydrogeological studies. The studies include mainly the characterization of aquifers’ functioning, the elaboration of the spring inventory, physicochemical analyses and geoconservation actions.This work is funded by the Courel Mountains UGGp and the project IGCP-730 Hydrogeological significance of Mediterranean geoparks, and supported by the International Geosciences Program of UNESCO.Peer reviewe

Design of a GIS-database for the management of the Courel Mountains UNESCO Global Geopark (Spain)

X Congreso Geológico de España, 5-7 Julio 2021, Vitoria - GasteizSe ha desarrollado una base de datos en un sistema de información geográfica (SIG) para la gestión del Geoparque Mundial de la UNESCO Montañas do Courel (NO de España). El SIG incluye 66 capas de información topográfica, geológica, minera, biológica, arqueológica y etnográfica, que pueden ser combinadas entre sí para elaborar mapas temáticos adaptados a la finalidad y al usuario. Los mapas generados son empleados en actividades de divulgación, en el diseño de cartografías técnicas de apoyo a los gestores del Geoparque, en el desarrollo de estudios científicos y en acciones de geoconservació