P2Y11 Receptor Antagonists
Journal of Medicinal Chemistry, 2005, Vol. 48, No. 22 7047
(7) Balogh, J.; Wihlborg, A. K.; Isackson, H.; Joshi, B. V.; Jacobson,
K. A.; Arner, A.; Erlinge, D. Phospholipase C and cAMP-
dependent positive inotropic effects of ATP in mouse cardiomyo-
cytes via P2Y(11)-like receptors. J. Mol. Cell Cardiol. 2005, 39,
223-230.
(8) van der Weyden, L.; Conigrave, A. D.; Morris, M. B. Signal
transduction and white cell maturation via extracellular ATP
and the P2Y11 receptor. Immunol. Cell Biol. 2000, 78, 369-
374.
(9) Wilkin, F.; Duhant, X.; Bruyns, C.; Suarez-Huerta, N.; Boey-
naems, J. M.; Robaye, B. The P2Y11 receptor mediates the ATP-
induced maturation of human monocyte-derived dendritic cells.
J. Immunol. 2001, 166, 7172-7177.
B3LYP and basis set 6-31G (d,p), using the Gaussian03
software package (Gaussian Inc., Pittsburgh, PA). Using these
parameters, a multiple linear regression analysis was con-
ducted using MOE 2004.03 (Chemical Computing Group Inc.,
Montreal, Canada).
Data Analysis of Intracellular Calcium Measure-
ments. Effects of single doses of antagonists (100 µM) were
expressed as a percentage of the agonist control responses.
Antagonist IC50 values (pIC50 ) -log IC50) represent the
concentration needed to inhibit by 50% the effect elicited by
single doses of agonists. Apparent functional Ki values (pKi )
-log Ki) were calculated according to the equation of Cheng
and Prusoff:38
(10) Communi, D.; Robaye, B.; Boeynaems, J. M. Pharmacological
characterization of the human P2Y11 receptor. Br. J. Pharmacol.
1999, 128, 1199-1206.
(11) Dunn, P. M.; Blakeley, A. G. Suramin:
a reversible P2-
Ki ) IC50/(1 + L/EC50)
purinoceptor antagonist in the mouse vas deferens. Br. J.
Pharmacol. 1988, 93, 243-245.
(12) Jacobson, K. A.; Jarvis, M. F.; Williams, M. Purine and pyrimi-
dine (P2) receptors as drug targets. J. Med. Chem. 2002, 45,
4057-4093.
where IC50 is the inhibitory concentration 50% of the antago-
nist, EC50 is the effective concentration 50% of the used
agonist, and L is the molar concentration of the used agonist.
IC50 values for antagonists and EC50 values for agonists were
derived from -log concentration - effect (inhibition) curves
fitted to the pooled data by logistic, nonlinear regression
analysis (Prism 4.00, GraphPad Software, San Diego, CA).
(13) Damer, S.; Niebel, B.; Czeche, S.; Nickel, P.; Ardanuy, U.;
Schmalzing, G.; Rettinger, J.; Mutschler, E.; Lambrecht, G.
NF279: a novel potent and selective antagonist of P2X receptor-
mediated responses. Eur. J. Pharmacol. 1998, 350, R5-R6.
(14) Soto, F.; Lambrecht, G.; Nickel, P.; Stuhmer, W.; Busch, A. E.
Antagonistic properties of the suramin analogue NF023 at
heterologously expressed P2X receptors. Neuropharmacology
1999, 38, 141-149.
(15) Hu¨lsmann, M.; Nickel, P.; Kassack, M.; Schmalzing, G.; Lam-
brecht, G.; Markwardt, F. NF449, a novel picomolar potency
antagonist at human P2X1 receptors. Eur. J. Pharmacol. 2003,
470, 1-7.
(16) Kassack, M. U.; Braun, K.; Ganso, M.; Ullmann, H.; Nickel, P.;
Boing, B.; Muller, G.; Lambrecht, G. Structure-activity relation-
ships of analogues of NF449 confirm NF449 as the most potent
and selective known P2X1 receptor antagonist. Eur. J Med.
Chem. 2004, 39, 345-357.
(17) Rettinger, J.; Braun, K.; Hochmann, H.; Kassack, M. U.;
Ullmann, H.; Nickel, P.; Schmalzing, G.; Lambrecht, G. Profiling
at recombinant homomeric and heteromeric rat P2X receptors
identifies the suramin analogue NF449 as a highly potent P2X-
(1) receptor antagonist. Neuropharmacology 2005, 48, 461-468.
(18) Nickel, P.; Haack, H. J.; Widjaja, H.; Ardanuy, U.; Gurgel, C.;
Duwel, D.; Loewe, H.; Raether, W. Potential filaricides. Suramin
analogs. Arzneimittelforschung. 1986, 36, 1153-1157.
(19) Corson, B. B.; Hazen, R. K. Organic Synthesis; Wiley: New York,
1943; pp 434-438.
Acknowledgment. This work was supported by
DFG (Deutsche Forschungsgemeinschaft) grants
GRK677/1 and GRK677/2 to H.U. and M.U.K. and
FOR450 (TP11) to G.S., and by the START program of
the Faculty of Medicine, RWTH Aachen University, to
R.H. S.M. was supported by a stipend provided by the
Bischo¨fliche Studienfo¨rderung Cusanuswerk. D.H. was
supported by a stipend from the DAAD (Deutscher
Akademischer Austauschdienst). We further acknowl-
edge the assistance of Mrs. M. Schneider for measuring
the mass spectra.
Appendix
Abbreviations: ATPγS: adenosine-5′-O-(3-thiotriph-
osphate); cAMP: cyclic 3′,5′-adenosinemonophosphate;
2-MeSADP: 2-methylthio-adenosine-5′-diphosphate;
PPADS: pyridoxal-5′-phosphate-6-azophenyl-2′,4′-di-
sulfonic acid; SEM: standard error of the mean.
(20) Harwood: L. M.; Moody, C. J.; Percy, J. M. Experimental Organic
Chemistry: Standard and Microscale; Blackwell Science (UK):
Oxford, 1999.
(21) Olson, E. S. A modification of the free radical bromination of
p-toluic acid. J. Chem. Educ. 1980, 57, 157.
(22) Gattermann, L.; Wieland, H. Die Praxis des organischen Chemik-
ers; Carbonsa¨urechloride und Sa¨ureanhydride; Walter de
Gruyter: Berlin; New York, 1982; p 304.
(23) Henecka, H. Methoden der Organischen Chemie: Carbonsa¨ure-
chloride aus Carbonsa¨uren/Einwirkung von Thionylchlorid auf
Carbonsa¨uren (Houben-Weyl); Georg Thieme Verlag: Stuttgart,
1952; pp 610-612.
(24) Muth, H.; Sauerbier, M. Methoden der Organischen Chemie:
Reduktion (Houben-Weyl); Georg Thieme Verlag: Stuttgart,
1980; pp 645-654.
(25) Kreimeyer, A.; Muller, G.; Kassack, M.; Nickel, P.; Gagliardi,
A. R. Suramin analogues with a 2-phenylbenzimidazole moiety
as partial structure; potential anti HIV- and angiostatic drugs,
2: Sulfanilic acid-, benzenedisulfonic acid-, and naphthalen-
etrisulfonic acid analogues. Arch. Pharm. (Weinheim) 1998, 331,
97-103.
(26) Kassack, M.; Nickel, P. Rapid, highly sensitive gradient narrow-
bore high-performance liquid chromatographic determination of
suramin and its analogues. J. Chromatogr. B Biomed. Appl.
1996, 686, 275-284.
(27) Communi, D.; Govaerts, C.; Parmentier, M.; Boeynaems, J. M.
Cloning of a human purinergic P2Y receptor coupled to phos-
pholipase C and adenylyl cyclase. J. Biol. Chem. 1997, 272,
31969-31973.
(28) Schachter, J. B.; Sromek, S. M.; Nicholas, R. A.; Harden, T. K.
HEK293 human embryonic kidney cells endogenously express
the P2Y1 and P2Y2 receptors. Neuropharmacology 1997, 36,
1181-1187.
(29) Yu, H.; Bianchi, B.; Metzger, R.; Lynch, K.; Kowaluk, E.; Jarvis,
M. F.; Van Biesen, T. Lack of specificity of [35S]-ATPgammaS
and [35S]-ADPbetaS as radioligands for ionotropic and metabo-
tropic P2 receptor binding. Drug Dev. Res. 2005, 48, 84-93.
Supporting Information Available: Synthetic proce-
dures and compound monographs (analytical data). This
material is available free of charge via the Internet at http://
pubs.acs.org.
References
(1) Burnstock, G. A basis for distinguishing two types of purinergic
receptor. In Cell membrane receptors for drugs and hormones:
a multidisciplinary approach; Straub, R. W., Bolis, L. Eds.;
Raven Press: New York, 1978; pp 107-118.
(2) Khakh, B. S.; Burnstock, G.; Kennedy, C.; King, B. F.; North,
R. A.; Seguela, P.; Voigt, M.; Humphrey, P. P. International
union of pharmacology. XXIV. Current status of the nomencla-
ture and properties of P2X receptors and their subunits. Phar-
macol. Rev. 2001, 53, 107-118.
(3) Abbracchio, M. P.; Boeynaems, J. M.; Barnard, E. A.; Boyer, J.
L.; Kennedy, C.; Miras-Portugal, M. T.; King, B. F.; Gachet, C.;
Jacobson, K. A.; Weisman, G. A.; Burnstock, G. Characterization
of the UDP-glucose receptor (re-named here the P2Y14 receptor)
adds diversity to the P2Y receptor family. Trends Pharmacol.
Sci. 2003, 24, 52-55.
(4) Muller, C. E. P2-pyrimidinergic receptors and their ligands.
Curr. Pharm. Des. 2002, 8, 2353-2369.
(5) Zhang, F. L.; Luo, L.; Gustafson, E.; Palmer, K.; Qiao, X.; Fan,
X.; Yang, S.; Laz, T. M.; Bayne, M.; Monsma, F., Jr. P2Y(13):
identification and characterization of a novel Galphai-coupled
ADP receptor from human and mouse. J. Pharmacol. Exp. Ther.
2002, 301, 705-713.
(6) Jacobson, K. A.; King, B. F.; Burnstock, G. Pharmacological
characterization of P2 (nucleotide) receptors. Celltransmissions
2000, 16, 3-16.