Skip to main content
PMC home page
British Journal of Pharmacology logoLink to British Journal of Pharmacology
. 1994 Aug;112(4):1151–1159. doi: 10.1111/j.1476-5381.1994.tb13204.x

Comparative studies on the affinities of ATP derivatives for P2x-purinoceptors in rat urinary bladder.

X Bo 1, B Fischer 1, M Maillard 1, K A Jacobson 1, G Burnstock 1
PMCID: PMC1910266  PMID: 7952876

Abstract

1. Radioligand binding assays have been used to determine the affinities of a series of ATP derivatives with modifications of the polyphosphate chain, adenine and ribose moieties of the ATP molecule for [H]-alpha,beta-methylene ATP ([3H]-alpha,beta-MeATP) binding sites in rat urinary bladder. 2. The replacement of the bridging oxygen in the triphosphate chain of ATP (pIC50 = 5.58) with a methylene or imido group markedly increased the affinity (691 fold in IC50 values for beta,gamma-imidoATP, 15 fold for beta,gamma-methylene ATP), and the replacement of an ionized oxygen on the gamma-phosphate with a sulphur (ATP gamma S) also led to increased affinity (5623 fold in IC50 values). 3. Modifications at N6, N1, and C-8 positions on the purine base usually reduced the affinity of ATP (a decrease of 2.8 fold in IC50 values for N6-methylATP and 8.9 fold for 8-bromo ATP), while the attachment of an alkylthio group to the C-2 position greatly increased the affinity for P2x-purinoceptors (from 3.5 to 98 fold increase in IC50 values). 4. Replacement of the 3'-hydroxyl group on the ribose with substituted amino or acylamino groups produced more potent P2x-purinoceptor agonists (an increase of 447 fold in IC50 values for 3'-deoxy-3'-benzylamino ATP and 28 fold for 3'-deoxy-3'-(4-hydroxyphenylpropionyl)amino ATP. 5. Diadenosine polyphosphates (Ap[n]A) were also shown to displace the [3H]-alpha,beta-MeATP binding. The rank order of potency was Ap6A > Ap5A > Ap4A >> Ap3A >> Ap2A.(ABSTRACT TRUNCATED AT 250 WORDS)

Full text

PDF
1154

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bo X. N., Burnstock G. High- and low-affinity binding sites for [3H]-alpha, beta-methylene ATP in rat urinary bladder membranes. Br J Pharmacol. 1990 Oct;101(2):291–296. doi: 10.1111/j.1476-5381.1990.tb12703.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bo X., Burnstock G. Heterogeneous distribution of [3H]alpha,beta-methylene ATP binding sites in blood vessels. J Vasc Res. 1993 Mar-Apr;30(2):87–101. doi: 10.1159/000158980. [DOI] [PubMed] [Google Scholar]
  3. Bo X., Burnstock G. Triphosphate, the key structure of the ATP molecule responsible for interaction with P2X-purinoceptors. Gen Pharmacol. 1993 May;24(3):637–640. doi: 10.1016/0306-3623(93)90223-k. [DOI] [PubMed] [Google Scholar]
  4. Brown C., Burnstock G., Cocks T. Effects of adenosine 5'-triphosphate (ATP) and beta-gamma-methylene ATP on the rat urinary bladder. Br J Pharmacol. 1979 Jan;65(1):97–102. doi: 10.1111/j.1476-5381.1979.tb17337.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bruns R. F. Adenosine receptor activation by adenine nucleotides requires conversion of the nucleotides to adenosine. Naunyn Schmiedebergs Arch Pharmacol. 1980;315(1):5–13. doi: 10.1007/BF00504224. [DOI] [PubMed] [Google Scholar]
  6. Burnstock G., Cusack N. J., Hills J. M., MacKenzie I., Meghji P. Studies on the stereoselectivity of the P2-purinoceptor. Br J Pharmacol. 1983 Aug;79(4):907–913. doi: 10.1111/j.1476-5381.1983.tb10535.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burnstock G., Kennedy C. Is there a basis for distinguishing two types of P2-purinoceptor? Gen Pharmacol. 1985;16(5):433–440. doi: 10.1016/0306-3623(85)90001-1. [DOI] [PubMed] [Google Scholar]
  8. Busse R., Ogilvie A., Pohl U. Vasomotor activity of diadenosine triphosphate and diadenosine tetraphosphate in isolated arteries. Am J Physiol. 1988 May;254(5 Pt 2):H828–H832. doi: 10.1152/ajpheart.1988.254.5.H828. [DOI] [PubMed] [Google Scholar]
  9. Busshardt E., Gerok W., Häussinger D. Regulation of hepatic parenchymal and non-parenchymal cell function by the diadenine nucleotides Ap3A and Ap4A. Biochim Biophys Acta. 1989 Feb 9;1010(2):151–159. doi: 10.1016/0167-4889(89)90155-9. [DOI] [PubMed] [Google Scholar]
  10. Choo L. K. The effect of reactive blue, an antagonist of ATP, on the isolated urinary bladders of guinea-pig and rat. J Pharm Pharmacol. 1981 Apr;33(4):248–250. doi: 10.1111/j.2042-7158.1981.tb13770.x. [DOI] [PubMed] [Google Scholar]
  11. Cusack N. J., Hourani S. M., Loizou G. D., Welford L. A. Pharmacological effects of isopolar phosphonate analogues of ATP on P2-purinoceptors in guinea-pig taenia coli and urinary bladder. Br J Pharmacol. 1987 Apr;90(4):791–795. doi: 10.1111/j.1476-5381.1987.tb11233.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dunn P. M., Blakeley A. G. Suramin: a reversible P2-purinoceptor antagonist in the mouse vas deferens. Br J Pharmacol. 1988 Feb;93(2):243–245. doi: 10.1111/j.1476-5381.1988.tb11427.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Edwards F. A., Gibb A. J., Colquhoun D. ATP receptor-mediated synaptic currents in the central nervous system. Nature. 1992 Sep 10;359(6391):144–147. doi: 10.1038/359144a0. [DOI] [PubMed] [Google Scholar]
  14. Evans R. J., Derkach V., Surprenant A. ATP mediates fast synaptic transmission in mammalian neurons. Nature. 1992 Jun 11;357(6378):503–505. doi: 10.1038/357503a0. [DOI] [PubMed] [Google Scholar]
  15. Fedan J. S., Hogaboom G. K., O'Donnell J. P. Further comparison of contractions of the smooth muscle of the guinea-pig isolated vas deferens induced by ATP and related analogs. Eur J Pharmacol. 1986 Oct 7;129(3):279–291. doi: 10.1016/0014-2999(86)90438-3. [DOI] [PubMed] [Google Scholar]
  16. Fedan J. S., Hogaboom G. K., Westfall D. P., O'Donnell J. P. Comparison of contractions of the smooth muscle of the guinea-pig vas deferens induced by ATP and related nucleotides. Eur J Pharmacol. 1982 Jul 9;81(2):193–204. doi: 10.1016/0014-2999(82)90437-x. [DOI] [PubMed] [Google Scholar]
  17. Fedan J. S., Lamport S. J. P2-purinoceptor antagonists. Ann N Y Acad Sci. 1990;603:182–197. doi: 10.1111/j.1749-6632.1990.tb37672.x. [DOI] [PubMed] [Google Scholar]
  18. Gordon J. L. Extracellular ATP: effects, sources and fate. Biochem J. 1986 Jan 15;233(2):309–319. doi: 10.1042/bj2330309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gough G. R., Maguire M. H., Satchell D. G. Three new adenosine triphosphate analogs. Synthesis and effects on isolated gut. J Med Chem. 1973 Oct;16(10):1188–1190. doi: 10.1021/jm00268a028. [DOI] [PubMed] [Google Scholar]
  20. Hoyle C. H., Chapple C., Burnstock G. Isolated human bladder: evidence for an adenine dinucleotide acting on P2X-purinoceptors and for purinergic transmission. Eur J Pharmacol. 1989 Dec 12;174(1):115–118. doi: 10.1016/0014-2999(89)90881-9. [DOI] [PubMed] [Google Scholar]
  21. Hoyle C. H., Edwards G. A. Activation of P1- and P2Y-purinoceptors by ADP-ribose in the guinea-pig taenia coli, but not of P2X-purinoceptors in the vas deferens. Br J Pharmacol. 1992 Oct;107(2):367–374. doi: 10.1111/j.1476-5381.1992.tb12753.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hoyle C. H., Knight G. E., Burnstock G. Suramin antagonizes responses to P2-purinoceptor agonists and purinergic nerve stimulation in the guinea-pig urinary bladder and taenia coli. Br J Pharmacol. 1990 Mar;99(3):617–621. doi: 10.1111/j.1476-5381.1990.tb12979.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hoyle C. H. Pharmacological activity of adenine dinucleotides in the periphery: possible receptor classes and transmitter function. Gen Pharmacol. 1990;21(6):827–831. doi: 10.1016/0306-3623(90)90440-w. [DOI] [PubMed] [Google Scholar]
  24. Lambrecht G., Friebe T., Grimm U., Windscheif U., Bungardt E., Hildebrandt C., Bäumert H. G., Spatz-Kümbel G., Mutschler E. PPADS, a novel functionally selective antagonist of P2 purinoceptor-mediated responses. Eur J Pharmacol. 1992 Jul 7;217(2-3):217–219. doi: 10.1016/0014-2999(92)90877-7. [DOI] [PubMed] [Google Scholar]
  25. Lukacsko P., Krell R. D. Response of the guinea-pig urinary bladder to purine and pyrimidine nucleotides. Eur J Pharmacol. 1982 Jun 4;80(4):401–406. doi: 10.1016/0014-2999(82)90086-3. [DOI] [PubMed] [Google Scholar]
  26. Lüthje J., Ogilvie A. The presence of diadenosine 5',5'''-P1,P3-triphosphate (Ap3A) in human platelets. Biochem Biophys Res Commun. 1983 Aug 30;115(1):253–260. doi: 10.1016/0006-291x(83)90997-x. [DOI] [PubMed] [Google Scholar]
  27. MacKenzie I., Kirkpatrick K. A., Burnstock G. Comparative study of the actions of AP5A and alpha,beta-methylene ATP on nonadrenergic, noncholinergic neurogenic excitation in the guinea-pig vas deferens. Br J Pharmacol. 1988 Jul;94(3):699–706. doi: 10.1111/j.1476-5381.1988.tb11578.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Michel A. D., Humphrey P. P. Distribution and characterisation of [3H]alpha,beta-methylene ATP binding sites in the rat. Naunyn Schmiedebergs Arch Pharmacol. 1993 Dec;348(6):608–617. doi: 10.1007/BF00167237. [DOI] [PubMed] [Google Scholar]
  29. Molineaux C. J., Lasdun A., Michaud C., Orlowski M. Endopeptidase-24.15 is the primary enzyme that degrades luteinizing hormone releasing hormone both in vitro and in vivo. J Neurochem. 1988 Aug;51(2):624–633. doi: 10.1111/j.1471-4159.1988.tb01084.x. [DOI] [PubMed] [Google Scholar]
  30. Munson P. J., Rodbard D. Ligand: a versatile computerized approach for characterization of ligand-binding systems. Anal Biochem. 1980 Sep 1;107(1):220–239. doi: 10.1016/0003-2697(80)90515-1. [DOI] [PubMed] [Google Scholar]
  31. Peterson G. L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346–356. doi: 10.1016/0003-2697(77)90043-4. [DOI] [PubMed] [Google Scholar]
  32. Pintor J., Torres M., Castro E., Miras-Portugal M. T. Characterization of diadenosine tetraphosphate (Ap4A) binding sites in cultured chromaffin cells: evidence for a P2y site. Br J Pharmacol. 1991 Aug;103(4):1980–1984. doi: 10.1111/j.1476-5381.1991.tb12363.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rapaport E., Zamecnik P. C. Presence of diadenosine 5',5''' -P1, P4-tetraphosphate (Ap4A) in mamalian cells in levels varying widely with proliferative activity of the tissue: a possible positive "pleiotypic activator". Proc Natl Acad Sci U S A. 1976 Nov;73(11):3984–3988. doi: 10.1073/pnas.73.11.3984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Satchell D. G., Maguire M. H. Inhibitory effects of adenine nucleotide analogs on the isolated guinea-pig taenia coli. J Pharmacol Exp Ther. 1975 Dec;195(3):540–548. [PubMed] [Google Scholar]
  35. Seifert R., Schultz G. Involvement of pyrimidinoceptors in the regulation of cell functions by uridine and by uracil nucleotides. Trends Pharmacol Sci. 1989 Sep;10(9):365–369. doi: 10.1016/0165-6147(89)90009-6. [DOI] [PubMed] [Google Scholar]
  36. Silinsky E. M., Gerzanich V., Vanner S. M. ATP mediates excitatory synaptic transmission in mammalian neurones. Br J Pharmacol. 1992 Aug;106(4):762–763. doi: 10.1111/j.1476-5381.1992.tb14408.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Stone T. W. Actions of adenine dinucleotides on the vas deferens, guinea-pig taenia caeci and bladder. Eur J Pharmacol. 1981 Oct 22;75(2-3):93–102. doi: 10.1016/0014-2999(81)90066-2. [DOI] [PubMed] [Google Scholar]
  38. Stone T. W., Paton D. M. Possible subtypes of ATP receptor producing contraction of rat vas deferens, revealed by cross-desensitisation. Gen Pharmacol. 1989;20(1):61–64. doi: 10.1016/0306-3623(89)90061-x. [DOI] [PubMed] [Google Scholar]
  39. Theobald R. J., Jr Subclasses of purinoceptors in feline bladder. Eur J Pharmacol. 1992 Dec 15;229(2-3):125–130. doi: 10.1016/0014-2999(92)90545-f. [DOI] [PubMed] [Google Scholar]
  40. Welford L. A., Cusack N. J., Hourani S. M. The structure-activity relationships of ectonucleotidases and of excitatory P2-purinoceptors: evidence that dephosphorylation of ATP analogues reduces pharmacological potency. Eur J Pharmacol. 1987 Sep 2;141(1):123–130. doi: 10.1016/0014-2999(87)90418-3. [DOI] [PubMed] [Google Scholar]
  41. Ziganshin A. U., Hoyle C. H., Bo X., Lambrecht G., Mutschler E., Bäumert H. G., Burnstock G. PPADS selectively antagonizes P2X-purinoceptor-mediated responses in the rabbit urinary bladder. Br J Pharmacol. 1993 Dec;110(4):1491–1495. doi: 10.1111/j.1476-5381.1993.tb13990.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from British Journal of Pharmacology are provided here courtesy of The British Pharmacological Society

RESOURCES