A fluorescent polarization-based assay for the identification of disruptors of the RCAN1/calcineurin A protein complex

5 pages, 4 figures, a table. 19891949 [PubMed]Calcineurin is a Ca(2+)/calmodulin-dependent serine/threonine protein phosphatase involved in many biological processes and developmental programs, including immune response. One of the most studied substrates of calcineurin is the transcription factor NFAT (nuclear factor of activated T cells) responsible for T-cell activation. Different anticalcineurin drugs, such as cyclosporine A and FK506, are the most commonly used immunosuppressants in transplantation therapies. Unfortunately, their mechanism of action, completely blocking the calcineurin phosphatase activity while also requiring continuous administration, bears severe side effects. During recent years, the family of regulators of calcineurin (RCAN) has been described and studied extensively as modulators of calcineurin signaling pathways. The RCAN1 region, spanning amino acids 198 to 218 and responsible for inhibiting the calcineurin-NFAT signaling pathway in vivo, has been identified. An RCAN1-derived peptide spanning this sequence interferes with the calcineurin-NFAT interaction without affecting the general calcineurin phosphatase activity. Here we report the development of an optimized in vitro high-throughput fluorescence polarization assay based on the disruption of the RCAN1(198-218)-CnA interaction for identifying molecules with immunosuppressant potential. This approach led us to identify dipyridamole as a disruptor of such interaction. Moreover, three small molecules with a potential immunosuppressive effect were also identifiedThis work was supported by grants from Fundació La Marató de TV3 (Ref. 030830), the Spanish Ministry of Education and Science (SAF2006-04815, BIO2004-00998, BIO2007-60066, CTQ2005-00995/BQU), the Fundación Mutua Madrileña 2007 and from the Generalitat de Catalunya (Ref. 2006 BE 00051)Peer reviewe

Inhibition of Sema-3A Promotes Cell Migration, Axonal Growth, and Retinal Ganglion Cell Survival

Semaphorin 3A (Sema-3A) is a secreted protein that deflects axons from inappropriate regions and induces neuronal cell death. Intravitreal application of polyclonal antibodies against Sema-3A prevents loss of retinal ganglion cells ensuing from axotomy of optic nerves. This suggested a therapeutic approach for neuroprotection via inhibition of the Sema-3A pathway.Funded by the EU seventh framework program, Grant Agreement #604884.Peer reviewe

The chemical and the discovery of drugs: a challenge and a necessity

7 pages, 6 figures.[CAT] En aquest article es fa un breu relat històric del paper del químic en el descobriment de fàrmacs, per endinsar-se després en les estratègies modernes que fan servir els científics d’avui per dissenyar i produir nous medicaments que ajudin a combatre les malalties que més preocupen la societat. Finalment, es dóna una panoràmica de la possible evolució del món del descobriment de fàrmacs, de la implicació del químic en aquest món i de la contribució de grups d’investigació del nostre Institut en aquest camp.[ENG] In this contribution, after a brief historical account on the role of chemists on drug discovery, a deeper insight modern strategies that scientists employ for the design and production of new drugs that could help to the diseases that concern our society is discussed. Thereafter, an overview on the potential evolution directions discovery linked to the valuable implications of chemists to assist in this evolution is given. Finally, a brief of the work that research groups from our Institute carry out in the Biomedicine area is also outlined.Suport econòmic del Ministeri de Ciència i Innovació (Projecte SAF 2008-00048) i de la Fundació Marató de TV3 (Projecte 070332).Peer reviewe

Nomenclatura de química orgànica. Secció H: composts modificats isotòpicament

29 pages.Peer reviewe

La química combinatòria al tombant de segle

10 pages, 7 figures.[CAT] La química combinatòria comprèn el disseny i la síntesi eficient i ràpida de col·leccions de composts (quimioteques) per ésser avaluats davant dianes seleccionades. Aquesta tecnologia ha contribuït al desenvolupament d’una metodologia sintètica molt variada (fase homogènia i heterogènia, automatització i robòtica, disseny in silico, tècniques analítiques, etc.). Els èxits aconseguits per la química combinatòria justifiquen l’impacte que ha assolit en els sectors acadèmic i industrial en aquest canvi de segle.Combinatorial chemistry deals with the efficient and rapid design and synthesis of collections of compounds (libraries) to be screened towards targets of selected significance. In addition, this technology has contributed to the development of different methodologies (synthesis in homogeneous and heterogeneous phase, automation and robotics, in silico design, analytical techniques, etc.). The success achieved by combinatorial chemistry accounts for its impact in academia and industry at the turn of this century.Volem fer constar la nostra gratitud a tots els integrants de la Unitat de Química Bioorgànica, i en particular al Dr. Jordi Bujons, a Marc Humet i a Núria Cortès, i així mateix als doctors Enrique Pérez-Payá (Universitat de València) i Antonio Ferrer- Montiel (Universitat Miguel Hernández) i als seus equips, per la tasca feta i els suggeriments, la confiança i el suport rebuts a l’hora d’embrancar-nos en l’aventura combinatòria i continuar mantenint el caliu inicial. Igualment agraïm els suport rebut de la CICYT (Projecte SAF98-0059), del Programa FEDER (2FD1997-1760-CO3-03), de la Fundació Marató TV3 (Projecte 2013/97), de DiverDrugs SL i de Merck Farma y Química.Peer reviewe

Potentialimplication of anilinederivatives in the ToxicOilSyndrome (TOS)

The ToxicOilSyndrome (TOS) was an epidemic disease appeared in central Spain in 1981, causing over 400 deaths and affecting more than 20,000 people, mainly women and children. The disease was linked to the consumption of rapeseed oil denatured with aniline, illegally refined at the ITH oil refinery in Seville, mixed with other oils and sold as edible olive oil. Among the anilinederivatives detected in the oil batches generated by an uncontrolled deodorisation procedure during the refining process, fatty acid anilides were first postulated as the causal agents. Nevertheless, compounds identified as 3-(N-phenylamino)propane-1,2-diol (PAP) and its mono-, di-, and triacyl derivatives (mPAP, dPAP and tPAP, respectively), were subsequently considered better biomarkers of toxicoils and the best candidates for causing the intoxication. In this account, we will discuss the results obtained in recent years by our group concerning: (a) The effect of different variables intervening in the deodorisation process that could influence the formation of PAP derivatives. To this end we decided to take the anilinederivatives linked to oleic acid as compound models since this is the fatty acid present in highest amounts in rapeseed oil. The study was focused on the influence of different parameters on the formation of the diester PAP derivative (OOPAP) the monoester derivative (OPAP) and the corresponding amide (oleanilide, OA), and the interactions between any two of these variables. Of particular interest was the interaction observed between OOPAP and OA, due to its potential relevance to the final composition of the toxicoil model. (b) Xenobiochemical aspects of PAP derivatives, specifically: the stereospecific hydrolysis of OPAP and OOPAP by human pancreatic lipase, the in vitro activation of PAP by human and rat liver microsomes as well as by recombinant 450 enzymes, and the formation and stability of GSH and N-acetylcysteine adducts of a highly reactive iminoquinone intermediate generated in the biotransformation of PAP.Financial support from the WHO TOS Committee is gratefully acknowledged.Peer reviewe

Potentialimplication of anilinederivatives in the ToxicOilSyndrome (TOS)

The ToxicOilSyndrome (TOS) was an epidemic disease appeared in central Spain in 1981, causing over 400 deaths and affecting more than 20,000 people, mainly women and children. The disease was linked to the consumption of rapeseed oil denatured with aniline, illegally refined at the ITH oil refinery in Seville, mixed with other oils and sold as edible olive oil. Among the anilinederivatives detected in the oil batches generated by an uncontrolled deodorisation procedure during the refining process, fatty acid anilides were first postulated as the causal agents. Nevertheless, compounds identified as 3-(N-phenylamino)propane-1,2-diol (PAP) and its mono-, di-, and triacyl derivatives (mPAP, dPAP and tPAP, respectively), were subsequently considered better biomarkers of toxicoils and the best candidates for causing the intoxication. In this account, we will discuss the results obtained in recent years by our group concerning: (a) The effect of different variables intervening in the deodorisation process that could influence the formation of PAP derivatives. To this end we decided to take the anilinederivatives linked to oleic acid as compound models since this is the fatty acid present in highest amounts in rapeseed oil. The study was focused on the influence of different parameters on the formation of the diester PAP derivative (OOPAP) the monoester derivative (OPAP) and the corresponding amide (oleanilide, OA), and the interactions between any two of these variables. Of particular interest was the interaction observed between OOPAP and OA, due to its potential relevance to the final composition of the toxicoil model. (b) Xenobiochemical aspects of PAP derivatives, specifically: the stereospecific hydrolysis of OPAP and OOPAP by human pancreatic lipase, the in vitro activation of PAP by human and rat liver microsomes as well as by recombinant 450 enzymes, and the formation and stability of GSH and N-acetylcysteine adducts of a highly reactive iminoquinone intermediate generated in the biotransformation of PAP.Financial support from the WHO TOS Committee is gratefully acknowledged.Peer reviewe

Uso de un compuesto para la elaboración de un medicamento destinado al tratamiento de una lesión producida por una reperfusión post-isquémica

Uso de un compuesto para la elaboración de un medicamento destinado al tratamiento de una lesión producida por una reperfusión post-isquémica. La presente invención se refiere al uso del compuesto CR-6 (3,4-dihidro-6-hidroxi-7-metoxi-2,2-dimetil-1(2H)- benzopirano) o de un compuesto derivado, para el tratamiento de una lesión producida en determinados individuos como consecuencia de la reperfusión tras un episodio de isquemia cerebral. Esta lesión se denomina lesión por reperfusión y se asocia a determinados signos precoces como es la hiperemia reactiva cuando ocurre la reperfusión post-isquémica. En la presente invención se demuestra que no todos los individuos que han sufrido isquemia van a responder favorablemente al tratamiento con este compuesto, sino solamente aquellos que presenten signos de lesión por reperfusión, donde dicho signo es hiperperfusión o hiperemia reactiva. La hiperperfusión se considera que tiene lugar cuando el flujo sanguíneo cerebral aumenta por encima de un 20% del valor basal, condición que no se da en todos los sujetos que han sufrido isquemia/reperfusión cerebral.Peer reviewedConsejo Superior de Investigaciones Científicas (España)A1 Solicitud de patente con informe sobre el estado de la técnic

Combinatorial chemistry in cancer research

10 pages, 6 figures.-- PMID: 17329219 [PubMed].Among the different strategies to treat cancer, chemotherapy approaches are the subject of intense research efforts. There is still a high demand for new anticancer drugs exhibiting improved efficiency and selectivity for their use in combined therapy strategies. The high development of molecular and cellular biology tools has made possible the set up of simple in vitro assays, susceptible to automation, thus bringing about the possibility of rapid screening of hundreds of compounds. Chemistry has reacted to this challenge by developing a new technology: combinatorial chemistry. By this procedure large collections of compounds, known as chemical libraries, can be prepared in a rapid and efficient manner. In recent years, combinatorial chemistry has had a great impact on drug discovery programmes addressed to tackling cancer pharmaceutical targets. In this review, the contribution of this technology to the discovery of anticancer drugs that are currently in clinical trials or already in the market is discussed.Financial support from Ministerio de Educación y Ciencia (Grants CTQ2005-00995 and GEN2003-20642-C09-09/NAC) and Fundació Marató TV3 (Grant 080830) is gratefully acknowledged.Peer reviewe

Generation of Quinoneimine Intermediates in the Bioactivation of 3-(N-Phenylamino)alanine (PAA) by Human Liver Microsomes: A Potential Link Between Eosinophilia-Myalgia Syndrome and Toxic Oil Syndrome.

7 paginas; 6 figuras.Eosinophilia-myalgia syndrome (EMS) was an intoxication episode that occurred in the US in 1989 and affected 1,500 people. EMS was associated with the ingestion of manufactured L-tryptophan, and 3-(N-phenylamino)alanine (PAA) was identified as one of the contaminants present in the L-tryptophan batches responsible for intoxication. In previous studies (Martínez-Cabot et al., Chem Res. Toxicol., in press), we have shown that the incubation of 3-(N-phenylamino)propane-1,2-diol (PAP), a toxic biomarker of the oil batches that caused Toxic Oil Syndrome in Spain, with human liver microsomes generates a reactive quinoneimine intermediate. The structural similarity between PAA and PAP led Mayeno and co-workers (Mayeno et al. (1995) Chem. Res. Toxicol. 8, 911–916) to hypothesize that both xenobiotics could be linked to a common etiologic agent. We thus set about to study the bioactivation of PAA by human liver microsomes. Under these conditions, PAA is converted to its 4′-hydroxy derivative, an unstable intermediate that is rapidly transformed into the final metabolites 4-aminophenol and formylglycine, which were identified in the incubations by GC/MS using the H2 18O-labeled medium. We also provide evidence that 4-aminophenol and formylglycine are formed from a quinoneimine intermediate via a pathway similar to that demonstrated for PAP bioactivation. This quinoneimine, in the absence of nucleophiles in the incubation medium, could isomerize to give the corresponding imine, which could undergo hydrolysis to yield the aforementioned final products. These findings establish that EMS and TOS are linked by a common toxic metabolite (4-aminophenol) and that they may be further linked by the concomitant release of potentially hazardous carbonyl species.Financial support from the WHO TOS Committee is gratefully acknowledged.Peer reviewe