3 pages, 3 schemes, 1 table.-- Available on line Dec 23, 2003. ; A solid-phase version of the Nozaki–Hiyama allylation of supported allylic bromides with aldehydes is described. α-Methylene γ-butyrolactones 7 can be obtained by cyclization cleavage of the supported intermediate homoallylic alcohols 6. ; Financial support from the European Union (contract FMRXCT98-0235), and DURSI Generalitat de Catalunya (2001SGR00342 and 2001SGR00085). ; Peer reviewed
Catecholamine-triggered β-adrenoceptor (β-AR) signaling is essential for the correct functioning of the heart. Although both β1 - and β2 -AR subtypes are expressed in cardiomyocytes, drugs selectively targeting β1 -AR have proven this receptor as the main target for the therapeutic effects of beta blockers in the heart. Here, we report a new strategy for the light-control of β1 -AR activation by means of photoswitchable drugs with a high level of β1 -/β2 -AR selectivity. All reported molecules allow for an efficient real-time optical control of receptor function in vitro. Moreover, using confocal microscopy we demonstrate that the binding of our best hit, pAzo-2, can be reversibly photocontrolled. Strikingly, pAzo-2 also enables a dynamic cardiac rhythm management on living zebrafish larvae using light, thus highlighting the therapeutic and research potential of the developed photoswitches. Overall, this work provides the first proof of precise control of the therapeutic target β1 -AR in native environments using light. ; We thank Ignacio Pérez (IQAC-CSIC, Barcelona), Yolanda Pérez (IQAC-CSIC, Barcelona), Lourdes Muñoz (SimChem, IQAC-CSIC, Barcelona) and Carme Serra (SimChem, IQAC-CSIC, Barcelona) for technical support. We thank Diana Baxter (Institute for Research in Biomedicine, IRB, Barcelona) for her thorough revision of the language of this manuscript. We thank Dr. Kees Jalink (The Netherlands Cancer Institute, Amsterdam, the Netherlands) for providing the plasmids encoding for the Epac-SH188 biosensor. We thank the University of Vic-Central University of Catalonia (UVic-UCC) and Dr. Marta Otero for the material assignment which helped in some biological assays. We thank Nikos Hatzakis for access to the Olympus IX81 confocal microscope (UCPH, DK). This work was supported by ERDF-FEDER European Fund and Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación (projects CTQ2017-89222-R and PID2020-120499RB-I00) and by the Catalan government (2017 SGR 1604) to A.L. X.R. research was financed by the ...
Background and Purpose Opioid-based drugs are the gold standard medicines for pain relief. However, tolerance and several side effects (i.e. constipation and dependence) may occur upon chronic opioid administration. Photopharmacology is a promising approach to improve the benefit/risk profiles of these drugs. Thus, opioids can be locally activated with high spatiotemporal resolution, potentially minimizing systemic-mediated adverse effects. Here, we aimed at developing a morphine photo-derivative (photocaged morphine), which can be activated upon light irradiation both in vitro and in vivo. Experimental Approach Light-dependent activity of pc-morphine was assessed in cell-based assays (intracellular calcium accumulation and electrophysiology) and in mice (formalin animal model of pain). In addition, tolerance, constipation and dependence were investigated in vivo using experimental paradigms. Key results In mice, pc-morphine was able to elicit antinociceptive effects, both using external light-irradiation (hind paw) and spinal cord implanted fibre-optics. In addition, remote morphine photoactivation was devoid of common systemic opioid-related undesired effects, namely, constipation, tolerance to the analgesic effects, rewarding effects and naloxone-induced withdrawal. Conclusion and Implications Light-dependent opioid-based drugs may allow effective analgesia without the occurrence of tolerance or the associated and severe opioid-related undesired effects. ; This work was supported by FEDER/Ministerio de Ciencia, Innovación y Universidades-Agencia Estatal de Investigación (SAF2017-87349-R, SAF2017-87199-R and CTQ2017-89222-R), ISCIII (PIE14/00034), and the Catalan government (2017 SGR 1604 and 2017 SGR 465). We thank Centres de Recerca de Catalunya (CERCA) Programme/Generalitat de Catalunya for IDIBELL institutional support. We thank Esther Castaño and Benjamín Torrejón from the Scientific and Technical Services (SCT) group at the Bellvitge Campus of the University of Barcelona for their technical assistance. We thank Lourdes Muñoz, Carme Serra and Juanlo Catena (SimChem, IQAC-CSIC, Barcelona) for support in the synthesis of compounds. Xavier Gasull and Aida Castellanos (Facultat de Medicina, Universitat de Barcelona) are acknowledged by their help with the obtention of DRG neurons from mouse. Also, we thank María Pilar Pérez Villamor (Laboratorios Dr. Esteve, Barcelona) for the kind gift of the HEK293 cell line permanently expressing μ receptors. ; Peer reviewed
The insecticide carbaryl is commonly found in indirectly exposed freshwater ecosystems at low concentrations considered safe for fish communities. In this study, we showed that after only 24 h of exposure to environmental concentrations of carbaryl (0.066-660 ng/L), zebrafish larvae exhibit impairments in essential behaviours. Interestingly, the observed behavioural effects induced by carbaryl were acetylcholinesterase-independent. To elucidate the molecular initiating event that resulted in the observed behavioural effects, in silico predictions were followed by in vitro validation. We identified two target proteins that potentially interacted with carbaryl, the α2B adrenoceptor (ADRA2B) and the serotonin 2B receptor (HTR2B). Using a pharmacological approach, we then tested the hypothesis that carbaryl had antagonistic interactions with both receptors. Similar to yohimbine and SB204741, which are prototypic antagonists of ADRA2B and HTR2B, respectively, carbaryl increased the heart rate of zebrafish larvae. When we compared the behavioural effects of a 24-h exposure to these pharmacological antagonists with those of carbaryl, a high degree of similarity was found. These results strongly suggest that antagonism of both ADRA2B and HTR2B is the molecular initiating event that leads to adverse outcomes in zebrafish larvae that have undergone 24 h of exposure to environmentally relevant levels of carbaryl. ; This work was supported by "Agencia Estatal de Investigación" from the Spanish Ministry of Science and Innovation (project PID2020-113371RB-C21), IDAEA-CSIC, Severo Ochoa Centre of Excellence (CEX2018-000794-S), which financed M.F. with Severo Ochoa funds. Juliette Bedrossiantz was supported by a PhD grant (PRE2018-083513) co-financied by the Spanish Government and the European Social Fund (ESF). This work was supported by Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación and ERDF-FEDER European Fund (projects CTQ2017-89222-R and PID2020-120499RB-I00) and by the Catalan Government (2017 SGR 1604 and 2017-SGR-1807) to AL. XR research was financed by the Spanish Ministry of Economy, Industry and Competitiveness (SAF2015-74132-JIN). We thank Dr. Kees Jalink (The Netherlands Cancer Institute, Amsterdam, the Netherlands) for providing the plasmids encoding for the Epac-SH188 biosensor and Dr. Karen Martinez (University of Copenhagen, Copenhagen, Denmark) for providing the HEK 293 SNAP-β1AR). The findings of this report are not to be construed as an official Department of the Army position unless so designated by other authorized documents. ; Peer reviewed
Metabotropic glutamate receptors (mGluRs) are dimeric G-protein-coupled receptors activated by the main excitatory neurotransmitter, L-glutamate. mGluR activation by agonists binding in the venus flytrap domain is regulated by positive (PAM) or negative (NAM) allosteric modulators binding to the 7-transmembrane domain (7TM). We report the cryo-electron microscopy structures of fully inactive and intermediate-active conformations of mGlu5 receptor bound to an antagonist and a NAM or an agonist and a PAM, respectively, as well as the crystal structure of the 7TM bound to a photoswitchable NAM. The agonist induces a large movement between the subunits, bringing the 7TMs together and stabilizing a 7TM conformation structurally similar to the inactive state. Using functional approaches, we demonstrate that the PAM stabilizes a 7TM active conformation independent of the conformational changes induced by agonists, representing an alternative mode of mGlu activation. These findings provide a structural basis for different mGluR activation modes. ; We would like to thank Joël Bockaert, Chris Tate, Philippe Marin, and Richard Henderson for careful reading and for their comments on the manuscript. We would like to thank SLS PSI XA06 microfocus beamline scientist (Villigen; Switzerland), MRC Laboratory Molecular Biology EM and computing core facilities (Cambridge, UK), and the platform of pharmacology Arpeges of the Institut de Génomique Fonctionnelle (Montpellier, France). This project was funded by the ATIP grant from CNRS. C.N. was funded by the Post-doctoral program from Montpellier University. A.E.B. was supported by the Labex EpiGenMed (program "Investissements d'avenir," ANR-10-LABX-12-01), L.B. was supported by Ligue Contre le Cancer. J.F.-I., X.G.-S., J.C., and A.L. were supported by Ministerio de Ciencia e Innovación, Agencia Estatal de Investigación and ERDF-FEDER European Fund (project CTQ2017-89222-R) and by the Catalan government (2017 SGR 1604). J.-L.B. was supported by ANR (ANR-17-CE11-0011). J.-P.P. was supported by the Fondation Recherche Médicale (FRM team: DEQ20170326522), grants from ANR (DynaMuR2, ANR-18-CE11-0004; Lanthslider, ANR-17-CE11-0046). G.C. and K.R.V. were supported by Medical Research Council grant MC-U105184322. K.R.V. acknowledges SERB, India for the Ramanujan Fellowship (RJN-094/2017), DBT B-Life grant DBT/PR12422/MED/31/287/2014, and the support of the Department of Atomic Energy, Government of India, Government of India, under Project Identification No. RTI4006. G.L. was supported by ANR grants AT2R-TRAAK Bioanalgesics (ANR-17-CE18-0001), DynaMuR2 (ANR-18-CE11-0004), SWITCH-ON (ANR-20-CE11-0019), and Fondation Recherche Médicale (ING20121226326). ; Peer reviewed
