title: Ponalrestat: a potent and specific inhibitor of aldose reductase.
abstract: Many of the complications of diabetes appear to be closely linked to increased conversion of tissue glucose to sorbitol which is catalysed by aldose reductase (aldehyde reductase 2, ALR2). Inhibition of ALR2 could, therefore, lead to a reduction in the development of diabetic complications. Ponalrestat ["Statil" (a trademark, the property of Imperical Chemical Industries PLC), "Prodiax" (a trademark, the property of Merck, Sharp and Dohme), ICI 128436, MK538] inhibits ALR2 from a number of sources. Until now, the mechanism of this inhibition has not been fully elucidated. In this paper, we present a detailed mechanism for inhibition of bovine lens ALR2 by ponalrestat. Treatment of humans with some ALR2 inhibitors leads to side-effects, some of which may result from interactions with other enzymes. Aldehyde reductase (ALR1) is probably the most closely related enzyme to ALR2. Inhibition of ALR1 from bovine kidney was, therefore, investigated in order to assess the specificity of ponalrestat. The values of Ki and Kies (apparent dissociation constants for inhibitor from enzyme-inhibitor and enzyme-inhibitor-substrate complexes, respectively) for the interactions of ponalrestat with ALR1 and ALR2 has been calculated by non-linear fitting of kinetic data. These values indicate that ponalrestat does not compete with binding of glucose of NADPH to ALR2, nor with binding of glucuronate or NADPH to ALR1. Lack of competition and the structural dissimilarity of substrates and inhibitor make it unlikely that ponalrestat will utilize substrate binding sites on other enzymes, and so produce undesirable side-effects via such a mechanism. Ponalrestat is a potent inhibitor (Ki = Kies = 7.7 nM) of ALR2 and follows a pure noncompetitive mechanism with respect to glucose. Efficacy, therefore, will not be decreased by development of hyperglycaemia. The compound is a mixed noncompetitive inhibitor of ALR1 when glucuronate is varied. The values of Ki and Kies are 60 microM and 3 microM, respectively, so that inhibition tends towards uncompetitive. The selectivity of ponalrestat in favour of ALR2, therefore, lies in the range 390 to 7,800-fold, being higher at lower concentrations of glucuronate. The high selectivity of ponalrestat in favour of ALR2 rather than ALR1 suggests that the compound is unlikely to inhibit other enzymes which have less homology with ALR2.
authors.affiliation: ICI Pharmaceuticals, Macclesfield, Cheshire, U.K.
authors.forename: W H
authors.initials: WH
authors.lastname: Ward
authors.forename: C M
authors.initials: CM
authors.lastname: Sennitt
authors.forename: H
authors.initials: H
authors.lastname: Ross
authors.forename: A
authors.initials: A
authors.lastname: Dingle
authors.forename: D
authors.initials: D
authors.lastname: Timms
authors.forename: D J
authors.initials: DJ
authors.lastname: Mirrlees
authors.forename: D P
authors.initials: DP
authors.lastname: Tuffin
mesh_terms: D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors*; D000818:Animals; D002417:Cattle; D004305:Dose-Response Relationship, Drug; D005947:Glucose / Q000494:pharmacology; D005965:Glucuronates / Q000494:pharmacology; D020723:Glucuronic Acid; D006943:Hyperglycemia / Q000201:enzymology; D007527:Isoenzymes / Q000037:antagonists & inhibitors; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D007908:Lens, Crystalline / Q000201:enzymology*; D010793:Phthalazines / Q000378:metabolism / Q000494:pharmacology*; D011485:Protein Binding; D011724:Pyridazines / Q000494:pharmacology*; D012984:Software; D013379:Substrate Specificity; D013401:Sugar Alcohol Dehydrogenases / Q000037:antagonists & inhibitors*
chemical_list: D005965:Glucuronates; D007527:Isoenzymes; D010793:Phthalazines; D011724:Pyridazines; C042829:ponalrestat; D020723:Glucuronic Acid; D013401:Sugar Alcohol Dehydrogenases; D000449:Aldehyde Reductase; D005947:Glucose