Identification of potential inhibitors of protein-protein interaction useful to fight against Ebola and other highly pathogenic viruses

16 p.-1 fig.-1 tab.Despite the efforts to develop new treatments against Ebola virus (EBOV) there is currently no antiviral drug licensed to treat patients with Ebola virus disease (EVD). Therefore, there is still an urgent need to find new drugs to fight against EBOV. In order to do this, a virtual screening was done on the druggable interaction between the EBOV glycoprotein (GP) and the host receptor NPC1 with a subsequent selection of compounds for further validation. This screening led to the identification of new small organic molecules with potent inhibitory action against EBOV infection using lentiviral EBOV-GP-pseudotype viruses. Moreover, some of these compounds have shown their ability to interfere with the intracellular cholesterol transport receptor NPC1 using an ELISA-based assay. These preliminary results pave the way to hit to lead optimization programs that lead to successful candidates.Funding from “la Caixa” Banking Foundation under the project code HR18-00469 is acknowledged. This research was partially supported through Instituto de Salud Carlos III (FIS PI 181007 and ISCIII-COV20/01007), CSIC (201980E024 and 202020E079), Spanish Ministry of Science and Innovation (RTI2018-097305-R-I00) and the European Commission Horizon 2020 Framework Programme (Project VIRUSCAN FETPROACT-2016 and VACDIVA-SFS-12-2019-1-862874).Peer reviewe

African swine fever virus-cell interactions: From virus entry to cell survival

Viruses have adapted to evolve complex and dynamic interactions with their host cell. The viral entry mechanism determines viral tropism and pathogenesis. The entry of African swine fever virus (ASFV) is dynamin-dependent and clathrin-mediated, but other pathways have been described such as macropinocytosis. During endocytosis, ASFV viral particles undergo disassembly in various compartments that the virus passes through en route to the site of replication. This disassembly relies on the acid pH of late endosomes and on microtubule cytoskeleton transport. ASFV interacts with several regulatory pathways to establish an optimal environment for replication. Examples of these pathways include small GTPases, actin-related signaling, and lipid signaling. Cellular cholesterol, the entire cholesterol biosynthesis pathway, and phosphoinositides are central molecular networks required for successful infection. Here we report new data on the conformation of the viral replication site or viral factory and the remodeling of the subcellular structures. We review the virus-induced regulation of ER stress, apoptosis and autophagy as key mechanisms of cell survival and determinants of infection outcome. Finally, future challenges for the development of new preventive strategies against this virus are proposed on the basis of current knowledge about ASFV-host interactions. © 2012 Elsevier B.V.WT075813 Wellcome Trust Foundation, UE EPIZONE FOOD-CT2006-016236;CSD2006-00007, AGL2009-09209; AGL2012-34533, Ministerio de Economía y CompetitividadPeer Reviewe

Identification of NPC1 as a novel SARS-CoV-2 intracellular target

Niemann-Pick type C1 (NPC1) receptor is an endosomal membrane protein that regulates intracellular cholesterol trafficking, which is crucial in the Ebola virus (EBOV) cycle. The severe acute respiratory syndrome coronavirus 2 (SARS- CoV-2) enters the cell by binding of the viral spike (S) protein to the ACE2 receptor. This requires S-protein processing either by the surface transmembrane serine protease TMPRSS2 for plasma membrane fusion or cathepsin L for endosomal entry. Additional host factors are required for viral fusion at endosomes. Here, we report a novel interaction of the SARS-CoV-2 nucleoprotein (N) with the cholesterol transporter NPC1. Moreover, small molecules interfering with NPC1 that inhibit EBOV entry, also inhibited human coronavirus. Our findings suggest an important role for NPC1 in SARS-CoV-2 infection, a common strategy shared with EBOV, and a potential therapeutic target to fight against COVID-19.This research was partially supported through “La Caixa” Banking Foundation (HR18-00469), Instituto de Salud Carlos III (ISCIII-COV20/01007), CSIC (201980E024 and 202020E079), Spanish Ministry of Science and Innovation (RTI2018-097305-R-I00) and the European Commission Horizon 2020 Framework Programme VACDIVA-SFS482 12-2019-1-862874.N

Immune responses against African swine fever virus infection

23 p.-2 fig.Infection with African swine fever virus (ASFV) leads to a short haemorrhagic course of disease that, depending on the virus isolate, results in up to 100% lethality in domestic and Eurasian wild pigs. Consequently, ASFV infection in swine is of considerable economic significance. This chapter explains the basics of antiviral immunity in swine, focusing on the ‘knowns’ and ‘unknowns’ of the response against ASFV. In particular, monocytes and macrophages play an essential role as the main targets of infection and are crucial in viral persistence and dissemination. Furthermore,ASFV has developed several mechanisms to influence the antiviral and cell biological activity of infected monocytes, including down-regulation of cell surface receptors (e.g. CD14 and MHC-I) and modulation of interferon and cytokine/chemokine responses. ASFV infected pigs also develop virus-specific antibodies that can be used diagnostically, and while the neutralising effect of these antibodies has led to their involvement in protective immunity being controversially discussed, they may still exhibit protective functions through complement-mediated lysis and/or antibody dependent cell-mediated cytotoxicity. Indeed, T cells (presumably CD8+) also play a central role in the elimination of the virus, as can be seen in experiments where, after depletion of these cells, pigs previously primed with an avirulent ASFV become ill, while non-depleted animals are protected from highly virulent challenge. Nonetheless, despite these advances in our knowledge, much remains unknown about antiviral immunity generated during the course of a natural ASFV infection, or in response to attenuated virus strains or immunisation. Although such studies would undoubtedly be technically challenging, a deeper understanding of the immunity developed by the natural hosts (i.e. bushpigs and warthogs) against ASFV infection would teach us a lot about an effective protection from ASFV infection, and the involvement of both the innate and adaptive immune systems in this process.This publication is based upon work from COST Action CA15116, ASF-STOP, supported by COST (European Cooperation in Science and Technology).Peer reviewe

Compuestos sulfurados derivados de fenilhidrazidas como agentes antivirales

La presente invención se refiere a derivados sulfurados de fenilhidrazidas de fórmula (1) como agentes antivirales. Estos compuestos, por tanto, son útiles para el tratamiento y/o prevención de enfermedades víricas, tales como, la enfermedad causada por el virus del Ébola y/o de la peste porcinaPeer reviewedConsejo Superior de Investigaciones Científicas (España), Fundación para la Investigación Biomédica del Hospital Universitario 12 de Octubre, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA)A1 Solicitud de patente con informe sobre el estado de la técnic

Compuestos sulfurados derivados de fenilhidrazidas como agentes antivirales

[EN] The present invention relates to to sulfated phenylhydrazide derivatives of formula (I) as antiviral agents. Said compounds are therefore useful for the prevention and/or treatment of viral diseases such as the disease caused by Ebola virus, swine fever or coronavirus[ES] La presente invención se refiere a derivados sulfurados de fenilhidrazidas de fórmula (I) como agentes antivirales. Estos compuestos, por tanto, son útiles para el tratamiento y/o prevención de enfermedades víricas, tales como, la enfermedad causada por el virus del Ébola, de la peste porcina o por coronavirus[FR] La présente invention concerne des dérivés sulfurés de phénylhydrazides de formule (I) comme agents antiviraux. Ces composés sont par conséquent utiles pour le traitement et/ou la prévention de maladies virales, telles que, la maladie causée par le virus Ebola, la peste porcine ou le coronavirusPeer reviewedConsejo Superior de Investigaciones Científicas (España), Fundación para la Investigación Biomédica del Hospital Universitario 12 de Octubre, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA)A1 Solicitud de patente con informe sobre el estado de la técnic

Operacions bàsiques al laboratori químic en xarxa. Una nova eina per a estudiants i professors

Un conjunt de professors de les facultats de Química i Farmàcia de la Universitat de Barcelona hem elaborat un material docent en suport electrònic, d"accés lliure a la xarxa, que descriu el procediment pràctic de diverses operacions bàsiques de treball al laboratori químic. L"objectiu principal és crear un material docent que serveixi de suport a l"aprenentatge dels estudiants i a la tasca docent del professorat involucrat en l"ensenyament del treball pràctic en l"etapa d"inici dels estudis universitaris

Pharmacological modulation of the interaction between tubulin and a SARS-CoV-2 protein

1p.-6 fig.The cytoskeleton is the main communication/transport route within cells and many viruses abuse on this cellular machine to fulfil their cycle. We initially identified the interaction of a SARS-CoV-2 protein with tubulin using a proteomic analysis. We next confirmed the interaction and identified the viral protein domain involved through in vitro co-immunoprecipitation assays and analytical ultracentrifugation experiments. Then, we focused on unveiling the molecular mechanism of the interaction to determine if the SARS-CoV-2 protein promote a stable microtubule assembly (as roads for motors) or induce microtubule dynamics (as main force generation for transport). We have combined biochemical, biophysical and structural studies to determine the ratio of protein-protein interaction and the resulting effect on tubulin assembly. We have found that this protein domain is able to promote microtubule depolymerization into rings and tubulin assembly into non-functional filaments likely because a preference for tubulin curved-conformation. Importantly, this effect is not dependent on nucleotide or nucleotide hydrolysis. Finally, tubulin is a well-known target in cancer diseases and there are four of the seven tubulin druggable sites exploited on chemotherapy. Hence, we have analyzed the ability of microtubule stabilizing (MSA) and destabilizing agents (MDA) on disrupting the interaction of SARS-CoV-2 protein with tubulin. We have found that MSAs keep microtubule structures even in the presence of the viral protein, while the effect of MDAs varies depending on their mechanism of action.This research work was funded by Ministerio de Ciencia e Innovación and CSIC. It was also funded by the European Commission – NextGenerationEU (Regulation EU2020/2094), through CSIC's Global Health Platform (PTI Salud Global).Peer reviewe

Com identificar compostos per cromatografia en capa prima

Es mostra la metodologia a seguir per desenvolupar una cromatografia de capa prima i la manipulació necessària per separar els components d'una mostra

Com utilitzar correctament la balança

S'introdueix a l'alumne en la metodologia bàsica per pesar amb una balança monoplat de baixa precisió (0,1 g)