Abstract
A chiral column high-performance liquid chromatographic method was developed for the assessment of the enantiomeric purity of the stereoisomers of N6-phenylisopropyladenosine (PIA). The observed chiral purity of R-PIA was greater than 99.9%, whereas S-PIA was found to contain 4.4% of the R-enantiomer. In radioligand binding studies, the observed affinity of S-PIA for the adenosine A1 receptor (IC50 240 nM) could entirely be attributed to its content of R-PIA (IC50 7.8 nM). Calculation of a theoretical IC50 of pure S-PIA for the A2 receptor yielded a value of 6700 nM, which was 35-fold higher than for R-PIA (190 nM). Concludingly, the utilization of enantiomeric impure S-PIA in the definition of adenosine receptor subclasses is questionable.
MeSH terms
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Adenosine / analogs & derivatives
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Adenosine / metabolism
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Animals
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Brain / metabolism
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Cell Membrane / drug effects
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Cell Membrane / metabolism
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Chromatography, High Pressure Liquid / methods*
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Phenethylamines / metabolism
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Phenylisopropyladenosine / chemistry
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Phenylisopropyladenosine / isolation & purification*
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Purinergic P1 Receptor Agonists
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Purinergic P1 Receptor Antagonists
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Rats
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Receptors, Purinergic P1 / classification*
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Receptors, Purinergic P1 / metabolism
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Stereoisomerism
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Xanthines / metabolism
Substances
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Phenethylamines
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Purinergic P1 Receptor Agonists
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Purinergic P1 Receptor Antagonists
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Receptors, Purinergic P1
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Xanthines
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2-(4-(2-carboxyethyl)phenethylamino)-5'-N-ethylcarboxamidoadenosine
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Phenylisopropyladenosine
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1,3-dipropyl-8-cyclopentylxanthine
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Adenosine