ABSTRACT
A cobalt-catalyzed annulation of the C(sp2)-H/N-H bond of indoloamides with alkynes assisted by 8-aminoquinoline is reported for the synthesis of six-membered indololactams. The use of salicylaldehyde as the ligand is crucial for this transformation. The protocol has a broad scope for both alkynes and indoles. Preparing an active Co complex illustrates that salicylaldehyde plays a key role in the C-H activation step. The synthetic applications are proven by the gram-scale reaction and one-step construction of the multicyclic 5-HT3 receptor antagonist.
Subject(s)
Aldehydes/chemistry , Alkynes/chemistry , Aminoquinolines/chemistry , Indoles/chemistry , Serotonin 5-HT3 Receptor Agonists/chemical synthesis , Amides/chemistry , Catalysis , Cobalt/chemistry , Molecular Structure , Serotonin 5-HT3 Receptor Agonists/chemistryABSTRACT
Introduction of minor variations to the substitution pattern of arylguanidine 5-hydroxytryptamine-3 (5-HT3) receptor ligands resulted in a broad spectrum of functionally-active ligands from antagonist to superagonist. For example, (i) introduction of an additional Cl-substituent(s) to our lead full agonist N-(3-chlorophenyl)guanidine (mCPG, 2; efficacy % = 106) yielded superagonists 7-9 (efficacy % = 186, 139, and 129, respectively), (ii) a positional isomer of 2, p-Cl analog 11, displayed partial agonist actions (efficacy % = 12), and (iii) replacing the halogen atom at the meta or para position with an electron donating OCH3 group or a stronger electron withdrawing (i.e., CF3) group resulted in antagonists 13-16. We posit based on combined mutagenesis, crystallographic, and computational analyses that for the 5-HT3 receptor, the arylguanidines that are better able to simultaneously engage the primary and complementary subunits, thus keeping them in close proximity, have greater agonist character while those that are deficient in this ability are antagonists.
Subject(s)
Guanidines/pharmacology , Serotonin 5-HT3 Receptor Agonists/pharmacology , Serotonin 5-HT3 Receptor Antagonists/pharmacology , Animals , Cell Line, Tumor , Dose-Response Relationship, Drug , Guanidines/chemical synthesis , Guanidines/chemistry , HEK293 Cells , Humans , Membrane Potentials/drug effects , Membrane Potentials/physiology , Mice , Models, Molecular , Molecular Structure , Mutation , Oocytes , Protein Binding , Receptors, Serotonin, 5-HT3/genetics , Receptors, Serotonin, 5-HT3/metabolism , Serotonin 5-HT3 Receptor Agonists/chemical synthesis , Serotonin 5-HT3 Receptor Agonists/chemistry , Serotonin 5-HT3 Receptor Antagonists/chemical synthesis , Serotonin 5-HT3 Receptor Antagonists/chemistry , XenopusABSTRACT
We have previously identified a novel class of 5-hydroxytryptamine type 3 receptor (5-HT3R) agonists sharing little structural similarity with orthosteric 5-HT3R ligands (Jørgensen et al., 2011). In the present study we have elucidated the functional characteristics and the mechanism of action of one of these compounds, trans-3-(4-methoxyphenyl)-N-(pentan-3-yl)acrylamide (TMPPAA). In electrophysiological recordings TMPPAA was found to be a highly-efficacious partial agonist equipotent with 5-HT at the 5-HT3A receptor (5-HT3AR) expressed in COS-7 cells and somewhat less potent at the receptor expressed in Xenopus oocytes. The desensitization kinetics of TMPPAA-evoked currents were very different from those mediated by 5-HT. Moreover, repeated TMPPAA applications resulted in progressive current run-down and persistent non-responsiveness of the receptor to TMPPAA, but not to 5-HT. In addition to its direct activation, TMPPAA potentiated 5-HT-mediated 5-HT3AR signalling, and the allosteric link between the two binding sites was corroborated by the analogous ability of 5-HT to potentiate TMPPAA-evoked responses. The agonism and potentiation exerted by TMPPAA at a chimeric α7-nACh/5-HT3A receptor suggested that the ligand acts through the transmembrane domain of 5-HT3AR, a notion further substantiated by its functional properties at chimeric and mutant human/murine 5-HT3ARs. A residue in the transmembrane helix 4 of 5-HT3A was identified as an important molecular determinant for the different agonist potencies exhibited by TMPPAA at human and murine 5-HT3ARs. In conclusion, TMPPAA is a novel allosteric agonist and positive allosteric modulator of 5-HT3Rs, and its aberrant signalling characteristics compared to 5-HT at the 5-HT3AR underline the potential in Cys-loop receptor modulation and activation through allosteric sites.
Subject(s)
Acrylamides/pharmacology , Mutant Chimeric Proteins/agonists , Phenyl Ethers/pharmacology , Receptors, Serotonin, 5-HT3/genetics , Serotonin 5-HT3 Receptor Agonists/pharmacology , Acrylamides/chemical synthesis , Allosteric Regulation , Allosteric Site , Animals , COS Cells , Chlorocebus aethiops , Evoked Potentials/drug effects , Evoked Potentials/physiology , Gene Expression , Kinetics , Mice , Mutant Chimeric Proteins/genetics , Mutant Chimeric Proteins/metabolism , Oocytes/cytology , Oocytes/drug effects , Oocytes/metabolism , Patch-Clamp Techniques , Phenyl Ethers/chemical synthesis , Protein Binding , Protein Structure, Secondary , Receptors, Serotonin, 5-HT3/metabolism , Serotonin/pharmacology , Serotonin 5-HT3 Receptor Agonists/chemical synthesis , Xenopus laevisABSTRACT
Serotonin type 3 (5-HT(3)) receptor partial agonists are being targeted as potential new drugs for the treatment of irritable bowel syndrome (IBS). Two new chemical series bearing indazole and indole cores have exhibited nanomolar binding affinity for the h5-HT(3)A receptor. A range of partial agonist activities in HEK cells heterologously expressing the h5-HT(3)A receptor were measured for the indazole series. Excellent 5-HT(3) receptor selectivity, favorable in vitro metabolic stability and CYP inhibition properties, and good oral in vivo potency in the murine von Bezold-Jarisch reflex model is exemplified thereby indicating the series to have potential utility as improved IBS agents.
