Skip to main content
SpringerLink
Log in

The gastrointestinal prokinetic benzamide derivatives are agonists at the non-classical 5-HT receptor (5-HT4) positively coupled to adenylate cyclase in neurons

  • Published:
Naunyn-Schmiedeberg's Archives of Pharmacology Aims and scope Submit manuscript

Summary

We have previously shown that a non-classical 5-hydroxytryptamine (5-HT4) receptor mediates the stimulation of adenylate cyclase activity in mouse embryo colliculi neurons in primary culture. The pharmacological characteristics of this receptor exclude the possibility that it belongs to the known 5-HT1, 5-HT2 or 5-HT3 receptor types. Here we report that this 5-HT receptor can be stimulated by 4-amino-5-chloro-2-methoxy substituted benzamide derivatives. All these compounds have been reported to be potent stimulants of gastrointestinal motility and some of them are 5-HT3 receptor antagonists. The rank order of potency of these substituted benzamide derivatives in stimulating cAMP formation was: cisapride > BRL 24924 > 5-HT > zacopride > BRL 20627 > metoclopramide. The non-additivity of benzamide and 5-HT activities suggests that 5-HT and the substituted benzamide derivatives act on the same receptor. Only ICS 205930, a recognized 5-HT3 receptor antagonist, competitively antagonized the stimulatory effect of cisapride, zacopride and BRL 24924. However, its pK i (6–6.3) for this new receptor was very different from its pK i for 5-HT3 receptors (pK i = 8 –10). Other selective 5-HT3 receptor antagonists with an indole group (BRL 43694 and GR 38032F), with a benzoate group (cocaïne, MDL 72222) or with a piperazine group (quipazine) were ineffective in reversing the stimulatory effect of benzamide derivatives. Exposure of neuronal cells to potent agonists at this receptor such as BRL 24924 rapidly reduces its capacity to stimulate cAMP production. For example, a preincubation of 10 min with BRL 24924 (100 μmol/l) reduced by 42% the ability of 5-HT to stimulate cAMP production. Cross-desensitization occurs between the effects of 5-HT and benzamides. The unique pharmacology of these nonclassical 5-HT receptors that we propose to call 5-HT4 is very close and even identical to the pharmacology of the high affinity 5-HT receptors involved in the indirect stimulation of smooth muscle in the guinea pig ileum. These receptors are different from the 5-HT3 receptors also present in guinea pig ileum.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Andrade R, Malenka RC, Nicoll RA (1986) A G protein couples serotonin and GABAB receptors to the same channels in hippocampus. Science 234:1261.

    Google Scholar 

  • Andrade R, Nicoll RA (1987) Pharmacologically distinct actions of serotonin on single pyramidal neurones of the rat hippocampus recorded in vitro. J Physiol 394:99–124.

    Google Scholar 

  • Arunlakshana O, Schild HO (1959) Some quantitative uses of drug antagonists. Br J Pharmacol 14:48–58.

    Google Scholar 

  • Barnes NM, Costall B, Naylor RJ (1988) 3H-zacopride: ligand for the identification of 5-HT3 recognition sites. J Pharmacol 40:548–551.

    Google Scholar 

  • Bockaert J, Nelson DL, Herbet A, Adrien J, Enjalbert A, Hamon M (1981) Serotonin receptors coupled with an adenylate cyclase in the rat brain: non identity with [3H]-5-HT binding sites. In: Haber B, Gabay S, Issidores MR, Alvisatos SGA (eds) Serotonin-Current aspects of neurochemistry and function. Plenum Press, New York, pp 327–345.

    Google Scholar 

  • Bockaert J, Dumuis A, Bouhelal R, Sebben M, Cory RN (1987) Piperazine derivatives including the putative anxiolytic drugs, buspirone and ipsapirone are agonists at 5-HT1A receptors negatively coupled with adenylate cyclase in hippocampal neurons. Naunyn-Schmiedeberg's Arch Pharmacol 335:588–592.

    Google Scholar 

  • Bouhelal R, Smounya L, Bockaert J (1988) 5-HT1B receptors are negatively coupled with adenylate cyclase in rat substantia nigra. Eur J Pharmacol 151:189–196.

    Google Scholar 

  • Bradley PB, Engel G, Feniuk W, Fozard JR, Humphrey PPA, Middlemiss DN, My Lecharane EJ, Richardson BP, Saxena PR (1986) Proposals for the classification and nomenclature of functional receptors for 5-hydroxytryptamine. Neuropharmacology 25:563–576.

    Google Scholar 

  • Branchek TA, Mawe GM, Gershon MD (1988) Characterization and localization of a peripheral neural 5-hydroxytryptamine receptor subtype (5-HT1P) with a selective agonist, 3H-5-hydroxyindalpine. J Neurosci 8:2582–2595.

    Google Scholar 

  • Buchheit KH, Engel G, Mutschler E, Richardson BP (1985) Study of the contractile effect of 5-hydroxytryptamine (5-HT) in the isolated longitudinal muscle strip from guinea pig ileum. Evidence for two distinct release mechanisms. Naunyn-Schmiedeberg's Arch Pharmacol 329:36–41.

    Google Scholar 

  • Chaffoy de Courcelles D, Leysen JE, De Clerck F, Van Belle H, Janssen PA (1985) Evidence that phospholipid turnover is the signal transducing system coupled to serotonin-S2 receptor sites. J Biol Chem 260:7603–7608.

    Google Scholar 

  • Cheng Y, Prusoff WH (1973) Relationship between the inhibition constant (K i) and the concentration of an inhibitor that causes a 50% inhibition (I50) of an enzymatic reaction. Biochem Pharmacol 22:3099–3108.

    Google Scholar 

  • Conn J, Sanders-Bush E (1985) Serotonin-stimulated phosphoinositide turnover: Mediation by the S2 binding site in rat cerebral cortex but not in subcortical regions. J Pharmacol Exp Ther 234:195–203.

    Google Scholar 

  • Conn J, Sanders-Bush E, Hoffman BJ, Hartig PR (1986) A unique serotonin receptor in choroid plexus is linked to phosphatidylinositol turnover. Proc Natl Acad Sci [USA] 83:4086–4088.

    Google Scholar 

  • Cory RN, Rouot B, Guillon G, Sladeczek F, Balestre MN, Bockaert J (1987) The 5-hydroxytryptamine (5-HT2) receptor stimulates inositol phosphate formation in intact and broken WRK1 cells: Determination of occupancy-response relationships for 5-HT agonists. J Pharmacol Exp Ther 241:258–267.

    Google Scholar 

  • Craig DA, Clarke DE (1989) 5-hydroxytryptamine and cholinergic mechanisms in guinea pig ileum. Br J Pharmacol 96:247P.

    Google Scholar 

  • De Vivo M, Maayani S (1986) Characterization of the 5-hydroxytryptamine1A receptor mediated inhibition of forskolin-stimulated adenylate cyclase activity in guinea pig and rat hippocampal membranes. J Pharmacol Exp Ther 238:248–253.

    Google Scholar 

  • Doyle VM, Creba JA, Rüegg UT, Hoyer D (1986) Serotonin increases the production of inositol phosphates and mobilizes calcium via the 5-HT2 receptor in A7r5 smooth muscle cells. Naunyn-Schmiedeberg's Arch Pharmacol 333:98–103.

    Google Scholar 

  • Dumuis A, Sebben M, Bockaert J (1988a) Pharmacology of 5-hydroxytryptamine1A receptors which inhibit cAMP production in hippocampal and cortical neurons in primary culture. Mol Pharmacol 33:178–186.

    Google Scholar 

  • Dumuis A, Bouhelal R, Sebben M, Cory R, Bockaert J (1988b) A non-classical 5-hydroxytryptamine receptor positively coupled with adenylate cyclase in the central nervous system. Mol Pharmacol 34:880–887.

    Google Scholar 

  • Dumuis A, Sebben M, Bockaert J (1989) BRL 24924: A potent agonist at a non-classical 5-HT receptor positively coupled with adenylate cyclase in colliculi neurons. Eur J Pharmacol 162: 381–384.

    Google Scholar 

  • Fargin A, Raymond JR, Lohse MJ, Kobilka BK, Caron MG, Lefkowitz RJ (1988) The genomic clone G-21 which resembles a β-adrenergic receptor sequence encodes the 5-HT1A receptor. Nature 335:358–360.

    Google Scholar 

  • Fozard JR (1987) 5-HT: The enigma variations. TIPS 8:501–506.

    Google Scholar 

  • Fozard JR (1989) Agonists and antagonists of 5-HT3 receptors. In: Saxena PR (ed) Cardiovascular pharmacology of 5-hydroxytryptamine: prospective therapeutic applications. Kluwer, Dordrecht, (in press).

    Google Scholar 

  • Gaddum JH, Picarelli ZP (1957) Two kinds of tryptamine receptors. Br J Pharmacol Chemother 12:323–328.

    Google Scholar 

  • Gozlan H, El Mestikawy S, Pichat L, Glowinsky J, Hamon M (1983) Identification of presynaptic serotonin autoreceptors using a new ligand: [3H]-PAT. Nature 305:140–142.

    Google Scholar 

  • Heuring RE, Peroutka SJ (1987) Characterization of a novel [3H]-5-hydroxytryptamine binding site subtype in bovine brain membranes. J Neurosci 7:894–903.

    Google Scholar 

  • Hoyer D, Engel G, Kalkman HO (1985) Characterization of the 5-HT1B recognition in rat brain: binding studies with (−)-[125I]-iodocyanopindolol. Eur J Pharmacol 118:1–12.

    Google Scholar 

  • Hoyer D, Neijt HC (1987) Identification of serotonin 5-HT3 recognition sites by radioligand binding in NG 108–15 neuroblastoma-glioma cells. Eur J Pharmacol 143:291–292.

    Google Scholar 

  • Hoyer D, Neijt HC (1988) Identification of 5-HT3 recognition sites in membranes of NIE-115 neuroblastoma cells by radioligand binding. Mol Pharmacol 33:303–309.

    Google Scholar 

  • Hoyer D, Schoeffter P (1988) 5-HT1D receptor-mediated inhibition of forskolin stimulated adenylate cyclase activity in calf substantia nigra. Eur J Pharmacol 147:145–147.

    Google Scholar 

  • Julius D, MacDermott AB, Axel R, Jessel TM (1988) Molecular characterization of a functional cDNA encoding the serotonin 1C receptor. Science 241:558–564.

    Google Scholar 

  • Kendall DA, Nahorski SR (1985) 5-hydroxytryptamine-stimulated inositol phospholipid hydrolysis in rat cerebral cortex slices: pharmacological characterization and effects of antidepressents. J Pharmacol Exp Ther 233:473–479.

    Google Scholar 

  • Kilpatrick GJ, Jones BJ, Tyers MB (1987) Identification and distribution of 5-HT3 receptors in rat brain using radioligand binding. Nature (Lond) 330:746–748.

    Google Scholar 

  • Leysen JE, Niemegeers CIE, Van Nueten JM, Laduron PM (1982) [3H]-ketanserin (R41468), a selective [3H]-ligand for serotonin2 receptor binding sites. Mol Pharmacol 21:301–314.

    Google Scholar 

  • Lubbert H, Hoffmann BJ, Snutch P, Van Dyke T, Levine JA, Hartig PR, Lester HA, Davidson N (1987) cDNA cloning of a serotonin 5-HT1C receptor by electrophysiological assays of mRNA injected Xenopus oocytes. Proc Natl Acad Sci [USA] 84:4332–4336.

    Google Scholar 

  • Markstein R, Hoyer D, Engel G (1986) 5-HT1A receptors mediate stimulation of adenylate cyclase in rat hippocampus. Naunyn-Schmiedeberg's Arch Pharmacol 333:335–341.

    Google Scholar 

  • Mawe GM, Branchek TA, Gerhon MD (1986) Peripheral neural serotonin receptors: Identification and characterization with specific antagonists. Proc Natl Acad Sci [USA] 83:9799–9803.

    Google Scholar 

  • Neijt HC, Karpf A, Schoefter P, Engel G, Hoyer D (1988) Characterisation of 5-HT3 recognition sites in membranes of NG 108–15 neuroblastoma-glioma cells with [3H]-ICS 205930. Naunyn-Schmiedeberg's Arch Pharmacol 337:493–499.

    Google Scholar 

  • Pazos A, Hoyer D, Palacios JM (1984) The binding of serotoninergic ligands to the porcine choroid plexus: Characterization of a new type of serotonin recognition site. Eur J Pharmacol 106: 539–546.

    Google Scholar 

  • Pedigo NW, Yamamura HI, Nelson DL (1981) Discrimination of multiple [3H]-5-hydroxytryptamine-binding sites by the neuroleptic spiperone in rat brain. J Neurochem 36:220–226.

    Google Scholar 

  • Peroutka SJ (1988) 5-hydroxytryptamine receptor subtypes: molecular, biochemical and physiological characterization. TINS 11:496–500.

    Google Scholar 

  • Peroutka SJ, Hamik A (1988) [3H]-quipazine labels 5-HT3 recognition sites in rat cortical membranes. Eur J Pharmacol 148:297–299.

    Google Scholar 

  • Pfeuffer-Friederich I, Kilbinder H (1984) Facilitation and inhibition by 5-hydroxytryptamine and R51619 of acetylcholine release from guinea pig myenteric neurons. In: Roman C (eds) Gastrointestinal Motility. MTP Press Ltd, pp 527–534.

  • Pritchett DB, Bach AWJ, Wozny M, Taleb O, Dal Toso R, Shih JC, Seeburg PH (1988) Structure and functional expression of cloned rat serotonin 5-HT2 receptor. EMBO J 7:4135–4140.

    Google Scholar 

  • Richardson BP, Engel G, Donatsch P, Stadler PA (1985) Identification of serotonin M-receptor subtypes and their specific blockade by a new class of drugs. Nature 316:126–131.

    Google Scholar 

  • Richardson BP, Buchheit KH (1988) The pharmacology, distribution and function of 5-HT3 receptors. In: Osborne NN, Hamon M (eds) Neuronal serotonin, vol 17. John Wiley and Sons Ltd, Chichester, pp 465–506.

    Google Scholar 

  • Sanger GJ (1987) Increased gut cholinergic activity and antagonism of 5-hydroxytryptamine M-receptors by BRL24924: potential clinical importance of BRL24924. Br J Pharmacol 91:77–87.

    Google Scholar 

  • Sanger GJ, Nelson DR (1989) Selective and functional 5-hydroxytryptamine3 receptor antagonism by BRL43694 (granisetron). Eur J Pharmacol 159:113–124.

    Google Scholar 

  • Sanger GJ, Wardle KA (1989) 5-hydroxytryptamine and cholinergic function in the gastrointestinal tract. In: Paoletti R, Vanhoutte PM (eds) Serotonin: from cell biology, pharmacology and therapeutics. Kluwer Academic Publishers, Dordrecht. (in press).

    Google Scholar 

  • Shoeffter P, Waeber C, Palacios JM, Hoyer D (1988) The 5-hydroxytryptamine 5-HT1D receptor subtype is negatively coupled to adenylate cyclase in calf substantia nigra. Naunyn-Schmiedeberg's Arch Pharmacol 337:602–608.

    Google Scholar 

  • Scholtysik G (1987) Evidence for inhibition by ICS 205930 and stimulation by BRL34915 of K+ conductance in cardiac muscle. Naunyn-Schmiedeberg's Arch Pharmacol 335:692–696.

    Google Scholar 

  • Schuurkes JAJ, Van Nueten JM, Van Deale PGH, Reyntjens AJ, Janssen PAJ (1985) Motor stimulating properties of cisapride on isolated gastrointestinal preparations on the guinea pig. J Pharmacol Exp Ther 234:775–783.

    Google Scholar 

  • Schuurkes JAJ, Megers AAHP, Niemegeers CJE, Leysen JE, Van Nueten JM (1987)A comparative study of the cholinergic vs the anti-dopaminergic properties of benzamides with gastrointestinal prokinetic activity. In: Szurzewski J (ed) Cellular physiology and clinical studies of gastrointestinal smooth muscle. Elsevier, Amsterdam, North Holland, pp 231–247.

    Google Scholar 

  • Shenker A, Maayani S, Weinstein H, Green JP (1987) Pharmacological characterization of two 5-hydroxytryptamine receptors coupled to adenylate cyclase in guinea pig hippocampal membranes. Mol Pharmacol 31:357–367.

    Google Scholar 

  • Van Heuven-Nolsen D (1988) 5-HT receptor subtype specific drugs and the cardiovascular system. TIPS 9:423–425.

    Google Scholar 

  • Waeber C, Schoefter P, Palacios JM, Hoyer D (1998a) Molecular pharmacology of 5-HT1D recognition sites: Radioligand bind ing studies in human, pig and calf brain membranes. Naunyn-Schmiedeberg's Arch Pharmacol 337:595–601.

    Google Scholar 

  • Waeber C, Dielt MM, Hoyer D, Probst A, Palacios JM (1988b) Visualization of a novel serotonin site 5-HT1D in human brain by autoradiography. Neurosci Lett 88:11–16.

    Google Scholar 

  • Waeber C, Dixon K, Hoyer D, Palacios JM (1988c) Localisation by autoradiography of neuronal 5-HT3 receptors in mouse CNS. Eur J Pharmacol 151:351–352.

    Google Scholar 

  • Watling K, Aspley S, Swain CJ, Saunders J (1988a) [3H]-Quaternised ICS205930 labels 5-HT3 receptor binding sites in rat brain. Eur J Pharmacol 149:397–398.

    Google Scholar 

  • Watling KJ (1988b) Radioligand binding studies identify 5-HT3 recognition sites in neuroblastoma cell lines and mammalian CNS. TIPS 9:227–229.

    Google Scholar 

  • Weiss S, Sebben M, Adolfo-Garcia-Sainz J, Bockaert J (1985) D2-dopamine receptor-mediated inhibition of cyclic AMP formation in striatal neurons in primary culture. Mol Pharmacol 27:595–599.

    Google Scholar 

  • Weiss S, Pin JP, Sebben M, Kemp DE, Sladeczek F, Gabrion J, Bockaert J (1986) Synaptogenesis of cultured striatal neurons in serum-free medium: A morphological and biochemical study. Proc Natl Acad Sci [USA] 83:2238–2242.

    Google Scholar 

  • Weiss S, Sebben M, Kemp D, Bockaert J (1986) Serotonin 5-HT1 receptors mediate inhibition of cyclic AMP production in neurons. Eur J Pharmacol 120:227–230.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Centre CNRS-INSERM de Pharmacologie-Endocrinologie, Rue de la Cardonille, F-34094, Montpellier Cedex 2, France

    Aline Dumuis, Michèle Sebben & Joël Bockaert

Authors
  1. Aline Dumuis
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michèle Sebben
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Joël Bockaert
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Send offprint requests to A. Dumuis at the above address

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dumuis, A., Sebben, M. & Bockaert, J. The gastrointestinal prokinetic benzamide derivatives are agonists at the non-classical 5-HT receptor (5-HT4) positively coupled to adenylate cyclase in neurons. Naunyn-Schmiedeberg's Arch Pharmacol 340, 403–410 (1989). https://doi.org/10.1007/BF00167041

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00167041

Key words

Search

Navigation