The Pharmacologic Estimation of Potencies of Agonists and Antagonists in the Classification of Adenosine Receptors

  • Terry P. Kenakin
  • H. J. Leighton


This chapter outlines some of the methods used to quantitate drug effects in isolated tissues for the purposes of adenosine receptor classification. There are advantages and disadvantages to studying adenosine receptors in isolated tissues. The disadvantages are that numerous pharmacokinetic factors and reflex mechanisms make extrapolation of isolated tissue data to whole animals and man hazardous. Also, the quantitative parameters for drugs are calculated from models assuming an equilibrium between the drugs and the receptors, which is a condition often not obtained in intact tissues. However, these factors are balanced by two major advantages. If it is assumed that the independent variable in pharmacologic experiments is drug concentration and the dependent variable is tissue response, experiments in isolated tissues allow adequate control of the independent variable (under careful experimental conditions) such that meaningful dependent variables can be obtained. Also, quantitative estimates of agonist efficacy, a quantity currently inaccessible in biochemical binding studies, can be obtained from experiments in isolated tissues. Thus, drug affinity and efficacy can be quantified and used in the classification of adenosine receptors.


Tissue Factor Adenosine Receptor Partial Agonist Full Agonist Equilibrium Dissociation Constant 


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  1. Arunlakshana, O., and Schild, H. O. 1959. Some quantitative uses of drug antagonists. Br. J. Pharmacol., 74: 48–58.Google Scholar
  2. Barlow, R. B., Scott, N. C., and Stephenson, R. P. 1967. The affinity and efficacy of onium salts on the frog rectus abdominis. Br. J. Pharmacol., 31. 188–196.Google Scholar
  3. Bender, A. S., Wu, P. H., and Phillis, J. W. 1981. Some biochemical properties of the rapid adenosine uptake system in rat brain synaptosomes. J. Neurochem., 57: 1282–1290.CrossRefGoogle Scholar
  4. Blinks, J. R. 1967. Evaluation of the cardiac effects of several beta adrenergic blocking agents. Ann. N.Y. Acad. Sci., 139 (3): 673–685.PubMedCrossRefGoogle Scholar
  5. Brown, C., Burnstock, G., and Cocks, T. 1979. Effects of adenosine 5’-triphosphate (ATP) and (3,7- methylene ATP on rat urinary bladder. Br. J. Pharmacol., 65: 97–102.PubMedCrossRefGoogle Scholar
  6. Bruns, R. F., Daly, J. W., and Snyder, S. H. 1980. Adenosine receptors in brain membranes: Binding of N6-cyclohexyl [3H]adenosine and l,3-diethyl-8-[3H]phenylxanthine. Proc. Natl. Acad. Sci., 77: 5547–5551.PubMedCrossRefGoogle Scholar
  7. Bruns, R. F., Daly, J. W., and Snyder, S. H. 1983. Adenosine receptor binding: Structure-activity analysis generates extremely potent xanthine antagonists. Proc. Natl. Acad. Sci., 80: 2077–2080.PubMedCrossRefGoogle Scholar
  8. Buckner, C. K., and Saini, R. K. 1975. On the use of functional antagonism to estimate dissociation constants for β-adrenergic receptor agonists in isolated guinea pig trachea. J. Pharmacol. Exp. Ther., 194: 565.PubMedGoogle Scholar
  9. Buckner, C. K., Torphy, T., and Costa, D. J. 1978. Studies on (3-adrenergic mediating changes in mechanical events and adenosine 3’, 5’-monophosphate levels. Rat atria. Europ. J. Pharmacol., 47: 259–211.CrossRefGoogle Scholar
  10. Burnstock, G. 1978. A basis for distinguishing two types of purinergic receptors. In: Cell Membrane Receptors for Drugs and Hormones, pp. 107–118. Ed. by Straub, R. S., and Bolis, L. Raven Press, New York.Google Scholar
  11. Burnstock, G., and Meghji, P. 1981. Distribution of Pr and P2-purinoceptors in guinea pig and frog heart. Br. J. Pharmacol., 73: 879–885.PubMedCrossRefGoogle Scholar
  12. Burnstock, G., Dumselay, B., and Smythe, A. 1972. Atropine resistant excitation of the urinary bladder: The possibility of transmission via nerves releasing a purine nucleotide. Br. J. Pharmacol., 44: 451–461.PubMedCrossRefGoogle Scholar
  13. Cass, C. E., Gaudette, L. A., and Paterson, A. R. P. 1974. Mediated transport of nucleosides in human erythrocytes. Specific binding of the inhibitor nitrobenzyl thioinosine to nucleoside transport sites in the erythrocyte membrane. Biochim. Biophys. Acta, 545: 1–10.Google Scholar
  14. Chaundry, I. H. 1982. Does ATP cross the plasma cell plasma membrane? Yale J. Biol. Med., 55: 1–10.Google Scholar
  15. Christie, J., and Satchell, D. G. 1980. Purine receptor in the trachea: Is there a receptor for ATP? Br. J. Pharmacol., 70: 512–514.PubMedCrossRefGoogle Scholar
  16. Clanachan, A. S., and Marshall, R. J. 1980. Potentiation of the effects of adenosine on isolated cardiac and smooth muscle by diazepam. Br. J. Pharmacol., 77: 459–466.CrossRefGoogle Scholar
  17. Clanachan, A. S., and Muller, M. J. 1980. Effect of adenosine uptake inhibition on the nature and potency of theophylline as a presynaptic adenosine receptor antagonist. Can. J. Physiol. Pharmacol., 55: 805–809.CrossRefGoogle Scholar
  18. Colman, R. A. 1980. Purine antagonists in the identification of adenosine receptors in guinea pig trachea and the role of purines in non-adrenergic inhibitory neurotransmission. Br. J. Pharmacol., 69: 359–366.CrossRefGoogle Scholar
  19. Colquhoun, D. 1973. The relation between classical and cooperative models for drug action. In: Drug Receptors, pp. 149–182. Ed. by Rang, H. P. University Park Press, Baltimore.Google Scholar
  20. Dahlen, S.-E., and Hedqvist, P. 1980. ATP, (3, 7-methylene ATP and adenosine inhibit non-cholinergic non-adrenergic transmission in rat urinary bladder. Acta Physiol. Scand., 709: 137–142.CrossRefGoogle Scholar
  21. Dowdle, E. B., and Maske, R. The effects of dipyridamole on guinea pig ileum longitudinal muscle- myenteric plexus preparation. Br. J. Pharmacol., 77: 235–244.Google Scholar
  22. Ebner, F. 1981. The inhibition by (±)-Propanolol of the positive inotropic effects of (±)-isoprenaline and (-)-noradrenaline, Naunyn-Schmeidehergs Arch. Pharmacol., 316: 96–101.Google Scholar
  23. Ebner, F., and Waud, D. R. 1978. The role of uptake of noradrenaline for its positive inotropic effect in relation to muscle geometry. Naunyn-Schmeidebergs Arch. Pharmacol., 303: 1–6.CrossRefGoogle Scholar
  24. Fedan, J. S., Hogaboom, G. K., O’Donnell, J. P., Colby, J., and Westfall, D. P. 1981. Contribution of purines to the neurogenic response of the vas deferens of the guinea pig. Europ. J. Pharmacol., 69: 41–53.Google Scholar
  25. Fedan, J. S., Hogaboom, G. K., Westfall, D. P., and O’Donnell, J. P. 1983. Photoaffinity labeling of P2-purinergic and H1-histamine receptors in smooth muscle. Proc. Fed. Am. Soc. Exp. Biol., 42: 2846–2850.Google Scholar
  26. Fredholm, B. B., Hedqvist, P., and Vernet, L. 1978. Effect of theophylline and other drugs on rabbit renal cyclic nucleotide phosphodiesterase, 5’-nucleotidase and adenosine deaminase. Biochem. Pharmacol., 27: 2845–2850.PubMedCrossRefGoogle Scholar
  27. Frew, R., And Baer, H. P. 1979. Adenosine-α, ß-methylene diphosphate effects in intestinal smooth muscle: Sites of action and possible prostaglandin involvement. J. Pharmacol. Exp. Ther., 277: 525–530.Google Scholar
  28. Frew, R., McKenzie, S. G., Bär, H. P., and Hutchison, K. J. 1976. The relaxant effects of adenosine- 5’-α, β-methylene diphosphonate on the longitudinal smooth muscle of the rabbit ileum. Can. J. Physiol. Pharmacol., 54: 626–629.PubMedCrossRefGoogle Scholar
  29. Furchgott, R. F. 1966. The use of β-haloalkylamines in the differentiation of receptors and in the determination of dissociation constants of receptor-agonist complexes. In: Advances in Drug Research, Volume 3, pp. 21–55. Ed. by Harper, N. J., and Simmonds, A. B. Academic Press, London.Google Scholar
  30. Furchgott, R. F. 1972. The classification of adrenoceptors (adrenergic receptors). An evaluation from the standpoint of receptor theory. In: Catecholamines Handbook of Experimental Pharmacology, Volume 33, pp. 283–335. Ed. by Bloschko, H., and Muscholl, E. Springer-Verlag, Berlin.Google Scholar
  31. Furchgott, R. F. 1978. Pharmacological characterization of receptors: Its relation to radioligand-bind- ing studies. Proc. Fed. Am. Soc. Exp. Biol., 37: 115–120.Google Scholar
  32. Furchgott, R. F., and Bursztyn, P. 1967. Comparison of dissociation constants and of relative efficacies of selected agonists acting on parasympathetic receptors. Ann. N.Y. Acad. Sci., 744: 882–889.CrossRefGoogle Scholar
  33. Furchgott, R. F., Jurkiewicz, A., and Jurkiewicz, N. H. 1973. Antagonism of propanolol to isopro- tererol in guinea pig trachea: Some cautionary findings. In: Frontiers in Catecholamine Research, pp. 295–300. Ed. by Usdin, E., and Snyder, S. H. Pergamon Press, Elmsford, New York.Google Scholar
  34. Gerlach, E., and Deuticke, B. 1963. Adenosine deaminase inhibition with dipyridamole. Arzneim- Forsch., 75: 48–54.Google Scholar
  35. Hoick, M. I., and Marks, B. H. 1978. Purine nucleoside and nucleotide interactions on normal and subsensitive alpha adrenoceptor responsiveness in guinea pig vas deferens. J. Pharmacol. Exp. Ther., 205: 104–117.Google Scholar
  36. Hopkins, S. V., and Goldie, R. G. 1971. A species difference in the uptake of adenosine by heart. Biochem. Pharmacol., 20: 3359–3365.Google Scholar
  37. Jager, L. P., and Schevers, J. A. M. 1980. A comparison of effects evoked in guinea pig taenia caecum by purine nucleotides and by purinergic nerve stimulation. J. Physiol., 299: 75–83.PubMedGoogle Scholar
  38. Jarvis, S. M., and Young, J. D. 1980. Nucleoside transport in human and sheep erythrocytes: Evidence that nitrobenzylthioinosine binds specifically to functional nucleoside transport sites. Biochem. J., 790: 377–383.Google Scholar
  39. Jenkinson, D. H. 1979. Partial agonists in receptor classification. In Proceeding of the VII Congress of Medicinal Chemistry, pp. 373–383. Ed. by Simpkins, M. A. Cotswold Press, Oxford..Google Scholar
  40. Jhamandas, K., Nakatsu, K., Downie, J. W., Bartlett, V., and Elliott, J. 1980. Action of coenzyme A on adenine derivative receptors in isolated tissues. Europ. J. Pharmacol., 62: 247–252.Google Scholar
  41. Karlsson, J.-A., Kjellin, G., and Persson, C. G. A. 1982. Effects on tracheal smooth muscle of adenosine and methylxanthines and their interaction. J. Pharm. Pharmacol., 54: 788–793.CrossRefGoogle Scholar
  42. Kaumann, A. J., and Marano, M. 1982. On equilibrium dissociation constants for complexes of drug- receptor subtypes. Selective and non-selective interactions of partial agonists with two plausible ß-adrenoeeptor subtypes mediating positive chronotropic effects of (-)-isoprenaline in kitten atria. Naunyn-Schmeidebergs Arch. Pharmacol., 575: 192–201.Google Scholar
  43. Kazic, T., and Milosavlgevic, D. 1980. Interaction between adenosine triphosphate and noradrenaline in the isolated vas deferens of the guinea pig. Br. J. Pharmacol., 77: 93–98.Google Scholar
  44. Kenakin, T. P. 1980a. Errors in the measurement of agonist potency ratios produced by uptake processes: A general model applied to fj-adrenoceptor agonists. Br. J. Pharmacol., 71:401–4X1.Google Scholar
  45. Kenakin, T. P. 1980b. Effects of equilibration time on the attainment of equilibrium between antagonists and drug receptors. Europ. J. Pharmacol., 66: 295–306.Google Scholar
  46. Kenakin, T. P. 1981. A pharmacological method to estimate the pKi of competitive inhibitors of agonist uptake processes in isolated tissues. Naunyn-Schmeidebergs Arch. Pharmacol., 316: 89–95.CrossRefGoogle Scholar
  47. Kertakin, T. P. 1982a. The potentiation of cardiac responses to adenosine by benzodiazepine. J. Pharmacol. Exp. Ther., 222: 752–758.Google Scholar
  48. Kenakin, T. P. 1982b. The Schild regression in the process of receptor classification. Can. J. Physiol. Pharmacol., 60: 249–265.CrossRefGoogle Scholar
  49. Kenakin, T. P. 1984. The classification of drugs and drug receptors in isolated tissues. Pharmacol. Rev., 36. 165-222.Google Scholar
  50. Kenakin, T. P., and Beek, D. 1980. Is prenalterol (HI33/80) really a selective beta-adrenoceptor agonist? Tissue selectivity resulting from differences in stimulus-response relationships. J. Phar-macol. Exp. Ther., 213: 406–412.Google Scholar
  51. Kenakin, T. P., and Black, J. W. 1978. The pharmacological classification of practolol and chloro- practolol. Mol. Pharmacol., 14: 601–623.Google Scholar
  52. Kolassa, N., Pfleger, K., and Tram, M. 1971. Species difference in action and elimination of adenosine after dipyridamole and hexobendine. Europ. J. Pharmacol., 13: 320–325.CrossRefGoogle Scholar
  53. Kukovetz, W. R., and Poch, G. 1970. Inhibition of cyclic-3’, 5’-nucleotide-phosphodiesterase as a possible mode of action of papavarine and similarly acting drugs. Naunyn Schmeidebergs Arch. Pharmacol., 267: 189–194.Google Scholar
  54. Leighton, H. J., and Parmeter, L. L. 1982. Schild analysis of purinergic receptors in various isolated tissues. Pharmacologist 24: 686.Google Scholar
  55. Lemoine, H., and Kaumann, A. J. 1983. A model for the interaction of competitive antagonists with two receptor-subtypes characterized by a Schild-plot with apparent slope unity. Naunyn Schmei-debergs Arch. Pharmacol., 322: 11–120.Google Scholar
  56. Maguire, M. H., and Satchell, D. G. 1979. The contribution of adenosine to the inhibitory actions of adenine nucleotides on the guinea pig taenia coli: Studies with phosphate-modified adenine nu-cleotide analogs and dipyridamole. J. Pharmacol. Exp. Ther., 2 /7: 626–633.Google Scholar
  57. MacKay, D. 1966a. A general analysis of the receptor-drug interaction. Br. J. Pharmacol., 26: 9–16.Google Scholar
  58. MacKay, D. 1966b. A new method for the analysis of drug-receptor interactions. In: Advances in Drug Research, Volume 3, pp. 1–20. Ed. by Harper, N. J., and Simmonds, A. B. Academic Press, London.Google Scholar
  59. Marangos, P. J., Patel, J., Clark-Rosenberg, R., and Martino, A. M. 1982. [3H]Nitrobenzylthioinosine binding as a probe for the study of adenosine uptake sites in brain. J. Neurochem., 39: 184–188.Google Scholar
  60. Marano, M., and Kaumann, A. J. 1976. On the statistics of drug-receptor constants for partial agonists. J. Pharmacol. Exp. Ther., 198: 518–525.Google Scholar
  61. Marcus, M. L., Skelton, C. L., Graver, L. E., and Epstein, S. E. 1972. Effects of theophylline on myocardial mechanics. Am. J. Physiol., 222: 1361–1365.PubMedGoogle Scholar
  62. Moritoki, H., Takei, M., Kasai, T., Matsumura, Y., and Ishida, Y. 1979. Possible involvement of prostaglandins in the action of ATP on guinea pig uterus. J. Pharmacol. Exp. Ther., 277: 104–111.Google Scholar
  63. Needleman, P., Minkes, M. S., and Douglas, J. R. 1974. Stimulation of prostaglandin biosynthesis by adenine nucleotides. Circ. Res., 34: 455–461.PubMedCrossRefGoogle Scholar
  64. O’Donnell, R. S., and Wanstall, J. C. 1979. The importance of choice of agonist in studies designed to predict (Pi adrenoceptor selectivity of antagonists from pA2 values on guinea pig trachea and atria. Naunyn-Schmeidebergs Arch. Pharmacol., 308. 183–190.Google Scholar
  65. Okwuasaba, F. K., Hamilton, J. T., and Cook, M. A. 1977. Relaxations of guinea pig fundic strip by adenosine, adenine nucleotides and electrical stimulation: Antagonism by theophylline and de- sensitization to adenosine and its derivatives. Europ. J. Pharmacol., 46: 181–198.CrossRefGoogle Scholar
  66. Parker, R. B. 1972. Measurement of drug-receptor dissociation constants of muscarinic agonists on intestinal smooth muscle. J. Pharmacol. Exp. Ther., 180: 62–10.PubMedGoogle Scholar
  67. Parker, R. B., and Waud, D. R. 1971. Pharmacological estimation of drug-receptor dissociation con-stants. Statistical evaluation. 1. Agonists. J. Pharmacol. Exp. Ther., 777: 1–12.Google Scholar
  68. Paterson, A. R. P., Jakobs, E. S., Harley, E. R., Fu, N.-W., Robins, M. J., and Cass, C. E. 1983. Inhibition of nucleoside transport In: Regulatory Function of Adenosine, Volume 2, pp. 203 - 220. Ed. by Berne, R. M., Rail, T. W., and Rubio, A. Martinus Nijhoff, Boston.CrossRefGoogle Scholar
  69. Paton, D. M. 1981. Structure-activity relations for presynaptic inhibition of noradrenergic and cholinergic transmission by adenosine: Evidence for action on Ai receptors. J. Auton. Pharmacol., 7: 287–290.CrossRefGoogle Scholar
  70. Phillis, J. W., Siemens, R. K., and Wu, P. H. 1980. Effects of diazepam on adenosine and acetylcholine release from rat cerebral cortex: Further evidence for a purinergic mechanism in action of di-azepam. Br. J. Pharmacol., 70: 341–348.CrossRefGoogle Scholar
  71. Plagemann, P. G. W., and Roth, M. F. 1969. Permeation as the rate limiting step in the phosphorylation of uridine and choline and their incorporation into macromolecules by Novikoff hepatoma cells. Biochemistry, 5: 4782–4789.CrossRefGoogle Scholar
  72. Renner, E. D., Plagemann, P. G. W., and Bernlohr, R. W. 1972. Permeation of glucose by simple and facilitated diffusion by Novikoff rat heptatoma cells in suspension culture and its relationship to glucose metabolism. J. Biol. Chem., 247: 5765–5776.PubMedGoogle Scholar
  73. Ruffolo, R. R. Jr., Dillard, R. D., Waddell, J. E., and Yaden, E. L. 1979. Receptor interactions of imidazolines. III. Structure-activity relationships governing alpha adrenergic receptor occupation and receptor activation of mono- and dimethoxy-substituted tolazoline derivatives in rat aorta. J. Pharmacol. Exp. Ther., 211: 133–738.Google Scholar
  74. Schild, H. O. 1947. pA, a new scale for the measurement of drug antagonism. Br. J. Pharmacol., 2: 189–206.Google Scholar
  75. Sjoberg, B., and Wahlstrom, B. A. 1975. The effort of ATP and related compounds on spontaneous mechanical activity in the rat portal vein. Acta Physiol. Scand., 94: 46–53.PubMedCrossRefGoogle Scholar
  76. Smellie, F. W., Davis, C. W., Daly, J. W., and Wells, J. N. 1979. Alkylxanthines: Inhibition of adenosine-elicited accumulation of cyclic AMP in brain slices and of brain phosphodiesterase activity. Life Sci., 24: 2475–2482.PubMedCrossRefGoogle Scholar
  77. Stafford, A. 1966. Potentiation of adenosine and the adenine nucleotides by dipyridamole. Br. J. Pharmacol., 25: 218–227.Google Scholar
  78. Stephenson, R. P. 1956. A modification of receptor theory. Br. J. Pharmacol., 77: 379–393.Google Scholar
  79. Su, Y. F., and Leighton, H. J. 1983. Functional antagonism to determine dissociation constants of presynaptic a-2adrenoceptor agonists. Pharmacologist, 25: 196.Google Scholar
  80. Thron, C. D. 1970. Graphical and weighted regression analyses for the determination of agonist dis-sociation constants. J. Pharmacol. Exp. Ther., 775: 541–553.Google Scholar
  81. Tsuzuki, J., and Newburgh, R. W. 1975. Inhibition of 5’-nucleotidase in rat brain by methylxanthines. J. Neurochem., 25: 895–896.PubMedCrossRefGoogle Scholar
  82. Van Belle, H. 1969. Uptake and deamination of adenosine by blood. Species differences, effect of pH, ions, temperature, and metabolic inhibitors. Biochim. Biophys. Acta, 792: 124–132.CrossRefGoogle Scholar
  83. Van Rossum, J. M. 1963. Cumulative dose-response curves. II. Technique for the making of dose- response curves in isolated organs and the evaluation of drug parameters. Arch. Int. Pharmacodyn. Ther., 745: 299–330.Google Scholar
  84. Venter, J. C. 1978. Cardiac sites of catecholamine action: Diffusion models for soluble and immobilized catecholamine action or isolated cat papillary muscles. Mol. Pharmacol., 74: 562–574.Google Scholar
  85. Waud, D. R. 1969a. A quantitative model for the effect of a saturable uptake on the slope of the dose- response curve. J. Pharmacol. Exp. Ther., 767: 140–141.Google Scholar
  86. Waud, D. R. 1969b. On the measurement of the affinity of partial agonists for receptors. J. Pharmacol. Exp. Ther., 770: 117–122.Google Scholar

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© Plenum Press, New York 1985

Authors and Affiliations

  • Terry P. Kenakin
    • 1
  • H. J. Leighton
    • 1
  1. 1.Department of Pharmacology The Wellcome Research LaboratoriesBurroughs Wellcome CompanyResearch Triangle ParkUSA

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