Skip to main content
SpringerLink
Log in

Selectivity of oxomemazine for the M1 muscarinic receptors

  • Research Articles
  • Published:
Archives of Pharmacal Research Aims and scope Submit manuscript

Abstract

The binding characteristics of pirenzepine and oxomemazine to muscarinic receptor were studied to evaluate the selectivity of oxomemazine for the muscarinic receptor subtypes in rat cerebral microsomes. Equilibrium dissociation constant (KD) of (−)-[3H]quinuclidinyl benzilate([3H]QNB) determined from saturation isotherms was 64 pM. Analysis of the pirenzepine inhibition curve of [3H]QNB binding to cerebral microsome indicated the presence of two receptor subtypes with high (Ki=16 nM, M1 receptor) and low (Ki=400 nM, M3 receptor) affinity for pirenzepine. Oxomemazine also identified two receptor subtypes with about 20-fold difference in the affinity for high (Ki=84nM, OH receptor) and low (Ki=1.65 μM, OL receptor) affinity sites. The percentage populations of M1 and M3 receptors to the total receptors were 61∶39, and those of OH and OL receptors 39∶61, respectively. Both pirenzepine and oxomemazine increased the KD value for [3H]QNB without affecting the binding site concentrations and Hill coefficient for the [3H]QNB binding, Oxomemazine had a 10-fold higher affinity at M1 receptors than at M3 receptors, and pirenzepine a 8-fold higher affinity at OH receptors than at OL receptors. Analysis of the shallow competition binding curves of oxomemazine for M1 receptors and pirenzepine for OL receptors yielded that 69% of M1 receptors were of OH receptors and the remaining 31% of OL receptors, and that 29% of OL receptors were of M1 receptors and 71% of M3 receptors. However, M3 for oxomemazine and OH for pirenzepine were composed of a uniform population. These results suggest that oxomemazine could be classified as a selective drug for M1 receptors and also demonstrate that rat cerebral microsomes contain three different subtypes of M1, M3 and the other site which is different from M1, M2 and M3 receptors.

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 Cited

  • Ambre, J. J., Bennett, D. R., Cranston, J. W., Dickinson, B. D., Evans, R. M., Glade, M. J., Kosman, M. E., Lampe, K. F., McCann, M. A., Pang, D. C., Proudfit, C. M., Rapoza, N. P., Ratko, T. A., Seidenfeld, J., Smith, S. J. and Weaver, R., Drug evaluations annual 1991.American Medical Association, pp. 1601–1604 (1991).

  • Barlow, R. B., Berry, K. J., Glenton, P. A. M., Nikolaou, N. M. and Soh, K. S., A comparison of affinity constants for muscarinic-sensitive acetylcholine receptors in guinea-pig atrial pacemaker cells at 29°C. and ileum at 37°C.Br. J. Pharmacol., 58, 613–620 (1976).

    PubMed  CAS  Google Scholar 

  • Barlow, R. B., Burston, K. N. and Vis, A., Three types of muscarinic receptors?Br. J. Pharmacol., 68, 141–142 (1980).

    Google Scholar 

  • Berrie, C. P., Birdsall, N. J. M., Burgen, A. S. V. and Hulme, E. C., The binding of muscarinic receptors in the lacrimal gland: Comparision with the cerebral cortex and myocardium.Br. J. Pharmacol., 78, 66P (1983).

    Google Scholar 

  • Birdsall, N. J. M. and Hulme, E. C., Biochemical studies on muscarinic acetylcholine receptors.J. Neurochem., 27, 7–16 (1976).

    Article  PubMed  CAS  Google Scholar 

  • Bolden, C., Cusack, B. and Richelson, E., Clozapine is a potent and selective muscarinic antagonist at the five cloned human muscarinic cholinergic receptors expressed in CHO-K1 cells.Eur. J. Pharmacol., 192, 205–206 (1991).

    Article  PubMed  CAS  Google Scholar 

  • Bolden, C., Cusack, B. and Richelson, E., Antagonism by antimuscarinic and neuroleptic compounds at the five cloned human muscarinic cholinergic receptors expressed in Chinese hamster ovary cells.J. Pharmacol. Exp. Ther., 260, 576–580 (1992).

    PubMed  CAS  Google Scholar 

  • Bonner, T. I., The molecular basis of muscarinic receptor density.Trends Neurol. Sci., 12, 148–151 (1989).

    Article  CAS  Google Scholar 

  • Bonner, T. I., Buckley, N. J., Young, A. C. and Brann, M. R., Identification of a family of muscarinic acetylcholine receptor genes.Science, 237, 527–532 (1987).

    Article  PubMed  CAS  Google Scholar 

  • Bonner, T. I., Young, A. C., Brann, M. R. and Buckley, N. J., Cloning and expression of the human and rat M5 muscarinic acetylcholine receptor genes.Neuron, 1, 403–410 (1988).

    Article  PubMed  CAS  Google Scholar 

  • Brown, D. A., Forward, A. and Marsh, S., Antagonist discrimination between ganglionic and ileal muscarinic receptors.Br. J. Pharmacol., 71, 362–364 (1980).

    PubMed  CAS  Google Scholar 

  • Buckley, N. J., Bonner, T. I. and Brann, M. R., Localization of a family of muscarinic receptor mRNAs in rat brain.J. Neurosci., 8, 4646–4652 (1988).

    PubMed  CAS  Google Scholar 

  • Buckley, N. J., Bonner, T. I., Buckley, C. M. and Brann, M. R., Antagonist binding properties of five cloned muscarinic receptors expressed in CHO-K1 cells.Mol. Pharmacol., 35, 469–476 (1989).

    PubMed  CAS  Google Scholar 

  • Cheng, Y. C. and Prusoff, W. H., Relationship between the inhibition constant (Ki) and the concentration of inhibitor which causes 50 percent inhibition (IC50) of an enzymatic reaction.Biochem. Pharmacol., 22, 3099–3108 (1973).

    Article  PubMed  CAS  Google Scholar 

  • Dauphin, F. and Hamel, E., Identification of multiple muscarinic binding site subtypes in cat and human cerebral vasculature.J. Pharmacol. Exp. Ther., 260, 660–667 (1992).

    PubMed  CAS  Google Scholar 

  • Doods, H. N., Mathy, M. J., Davidesko, D., Van, Charldorp, K. J., De, Jonge, A. and Van, Zwieten, P. A., Selectivity of muscarinic antagonists in radioligand and in vivo experiments for the putative M1, M2 and M3 receptors.J. Pharmacol. Exp. Ther., 242, 257–262 (1987).

    PubMed  CAS  Google Scholar 

  • Delmendo, R. E., Michel, A. D. and Whiting, R. L., Affinity of muscarinic receptor antagonists for three putative muscarinic receptor binding sites.Br. J. Pharmacol., 96, 457–464 (1989).

    PubMed  CAS  Google Scholar 

  • Dörje, F., Wess, J., Lambrecht, G., Tacke, R., Mutschler, E. and Brann, M. R., Antagonist binding profiles of five cloned human muscarinic receptor subtypes.J. Pharmacol. Exp. Ther., 256, 727–733 (1991).

    PubMed  Google Scholar 

  • El-Fakahany, E. E., Cioffi, C. L., Abdellatif, M. M. and Miller, M. M., Competitive interaction of pirenzepine with rat brain muscarinic acetylcholine receptors.Eur. J. Pharmacol., 131, 237–247 (1986).

    Article  PubMed  CAS  Google Scholar 

  • Giachetti, A., Micheletti, R. and Montagna, E., Cardioselective profile of AF-DX 116, a muscarinic M2 receptor antagonist.Life Sci., 38, 1663–1672 (1986).

    Article  PubMed  CAS  Google Scholar 

  • Giraldo, E., Hammer, R. and Ladinsky, H., Distribution of muscarinic receptor subtypes in rat brain as determined in binding studies with AF-DX 116 and pirenzepine.Life Sci., 40, 833–840 (1987).

    Article  PubMed  CAS  Google Scholar 

  • Goyal, R. K. and Rattan, S., Neurohumoral, hormonal, and drug receptors for the lower esophageal sphincter.Gastroenterology, 74, 598–618 (1978).

    PubMed  CAS  Google Scholar 

  • Hammer, R., Muscarinic receptors in the stomach.Scand. J. Gastroenterol., 15 (Suppl. 66), 5–11 (1980).

    CAS  Google Scholar 

  • Hammer, R., Berrie, C. P., Birdsall, N. J. M., Burgen, A. S. V. and Hulme, E. C., Pirenzepine distinguishes between different subclasses of muscarinic receptors.Nature (Lond.), 283, 90–92 (1980).

    Article  CAS  Google Scholar 

  • Hammer, R. and Giachetti, A., Muscarinic receptor subtypes: M1 and M2 biochemical and functional characterization.Life. Sci., 31, 2991–2998 (1982).

    Article  PubMed  CAS  Google Scholar 

  • Hammer, R. and Giachetti, A., Selective muscarinic receptor antagonists.Trends Pharmacol. Sci., 5, 18–20 (1984).

    Article  CAS  Google Scholar 

  • Hammer, R., Giraldo, E., Schiavi, G. B., Monferini, E. and Ladinsky, H., Binding profile of a novel cardioselective muscarinic receptor antagonist, AF-DX 116, to membrane of peripheral tissues and brain in the rat.Life Sci., 38, 1653–1662 (1986).

    Article  PubMed  CAS  Google Scholar 

  • Hirschowitz, B. I. R., Hammer, R., Giachetti, A., Kierns, J. J. and Levine, R. R., Subtypes of muscarinic receptors.Trends Pharmacol. Sci., 5 (suppl.), pp. VII (1984).

    Google Scholar 

  • Kloog, Y., Egozi, Y. and Sokolovskey, M., Characterization of muscarinic acetylcholine receptors from mouse brain: evidence for regional heterogeneity and isomerization.Mol. Pharmacol., 15, 545–548 (1979).

    PubMed  CAS  Google Scholar 

  • Lazareno, S. and Roberts, F. F., Functional and binding studies with muscarinic M2 subtype selective antagonists.Br. J. Pharmacol., 98, 309–317 (1989).

    PubMed  CAS  Google Scholar 

  • Lee, S. W., Park, Y. J., Park, I. S. and Lee, J. S., Interaction of antihistaminics with muscarinic receptor (II): Action on the cerebral muscarinic M1 receptor.Yakhak Hoeji, 34(4), 224–237 (1990).

    Google Scholar 

  • Lee, S. W. and Kim, J. G., Binding profiles of oxomemazine to the muscarinic receptor subtypes.Korean J. Pharmacol., 30, 49–57 (1994).

    CAS  Google Scholar 

  • Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R. J., Protein measurement with the Folin phenol reagent.J. Biol. Chem., 193, 265–275 (1951).

    PubMed  CAS  Google Scholar 

  • Luthin, G. R. and Wolfe, B. B., Comparison of [3H] pirenzepine and [3H]quinuclidinylbenzilate binding to muscarinic cholinergic receptors in rat brain.J. Pharmacol. Exp. Ther., 228, 648–655 (1984).

    PubMed  CAS  Google Scholar 

  • Micheletti, R., Montagna, E. and Giachetti, A., AF-DX 116, a cardioselective muscarinic antagonist.J. Pharmacol. Exp. Ther., 241, 628–634 (1987).

    PubMed  CAS  Google Scholar 

  • Michel, A. D. and Whiting, R. L., Methoctramine reveals heterogeneity of M2 muscarinic receptors in longitudinal ileal smooth muscle membranes.Eur. J. Pharmacol., 145, 305–311 (1988).

    Article  PubMed  CAS  Google Scholar 

  • Munson, P. J. and Rodbard, D., LIGAND: A versitile computerized approach for characterization of ligand-binding systems.Anal. Biochem., 107, 220–239 (1980).

    Article  PubMed  CAS  Google Scholar 

  • Peralta, E. G., Ashkenazi, A., Winslow, J. W., Smith, D. H., Ramachandran, J. and Caron, D., Distinct primary structures, ligand binding properties and tissue specific expression of four human muscarinic acetylcholine receptors.EMBO J., 6, 3923–3929 (1987).

    PubMed  CAS  Google Scholar 

  • Scatchard, G., The attractions of proteins for small molecules and ions.Ann. N. Y. Acad. Sci., 51, 660–672 (1949).

    Article  CAS  Google Scholar 

  • Watson, M., Yamamura, H. I. and Roeske, W. R., A unique regulatory profile and regional distribution of [3H]pirenzepine binding in the rat provide evidence for distinct M1 and M2 muscarinic receptor subtypes.Life Sci., 32, 3001–3011 (1983).

    Article  PubMed  CAS  Google Scholar 

  • Wess, J., Lambrecht, G., Mutschler, E., Brann, M. R. and Dörje, F., Selectivity profile of the novel muscarinic antagonist UH-AH 37 determined by the use of cloned receptors and isolated tissue preparations.Br. J. Pharmacol., 102, 246–250 (1991).

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. College of Pharmacy, Yeungnam University, 712-749, Gyongsan, Korea

    Shin Woong Lee, Chang Woo Woo & Jeung Gu Kim

Authors
  1. Shin Woong Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Chang Woo Woo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jeung Gu Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lee, S.W., Woo, C.W. & Kim, J.G. Selectivity of oxomemazine for the M1 muscarinic receptors. Arch. Pharm. Res. 17, 443–451 (1994). https://doi.org/10.1007/BF02979123

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02979123

Key words

Search

Navigation