ABSTRACT
Aspirin irreversibly inhibits cyclooxygenase (COX) by acetylating a serine residue in the active site. We synthesized a series of novel acylating agents based on our previously reported acetylating compound, O-acetylsalicylhydroxamic acid. One of these, triacetylsalicylhydroxamic acid (TriAcSHA) was more effective than aspirin and O-acetylsalicylhydroxamic acid in inactivating both COX-1 and COX-2. Preincubation of COX-1 with inhibitor for 5 min yielded IC(50) values of 18 microM for TriAcSHA and 60 microM for acetylsalicylic acid. Inhibition was time-dependent, with complete inhibition within 10 min at a concentration of 50 microM. As with aspirin, mutation of the serine 530 of COX-1 to alanine abolished the activity of the TriAcSHA. Mutation of the alanine 119 to a glutamine markedly reduced the sensitivity to TriAcSHA, suggesting that this residue was necessary for the interaction with the enzyme. TriAcSHA was also more effective than aspirin as an inhibitor of platelet aggregation induced by arachidonic acid. The diacetylated phenylhydroxamates N-methyl-O,O-diacetylsalicylhydroxamic acid, N,O-diacetylbenzohydroxamic acid, and 2-methyl-O,N-diacetylbenzohydroxamic acid showed reduced or absent activity against COX-1. In addition, we synthesized a series of triacylsalicylhydroxamic acids with progressively longer acyl groups (three to six carbons). All of the compounds inhibited COX-1 and demonstrated progressively greater COX-1 selectivity with increasing number of carbons. Hence, salicylhydroxamic acid provides a versatile backbone for the generation of a family of acylating inhibitors of cyclooxygenase.
Subject(s)
Cyclooxygenase Inhibitors/chemical synthesis , Isoenzymes/antagonists & inhibitors , Salicylamides/chemical synthesis , Acetylation , Animals , Arginine/genetics , Arginine/metabolism , Binding Sites , COS Cells , Cyclooxygenase 1 , Cyclooxygenase 2 , Cyclooxygenase 2 Inhibitors , Cyclooxygenase Inhibitors/chemistry , Cyclooxygenase Inhibitors/pharmacology , Humans , Hydroxamic Acids/chemistry , Isoenzymes/genetics , Membrane Proteins , Platelet Aggregation/drug effects , Prostaglandin-Endoperoxide Synthases/genetics , Salicylamides/chemistry , Salicylamides/pharmacology , Serine/genetics , Serine/metabolismABSTRACT
Aspirin is unique among clinically used nonsteroidal antiinflammatory drugs in that it irreversibly inactivates prostaglandin (PG) H2 synthase (PGHS) via acetylation of an active-site serine residue. We report the synthesis and characterization of a novel acetylating agent, O-acetylsalicylhydroxamic acid (AcSHA), which inhibits PGE2 synthesis in vivo and blocks the cyclooxygenase activity of PGHS in vitro. AcSHA requires the presence of the active-site residue Ser-529 to be active against human PGHS-1; the S529A mutant is resistant to inactivation by the inhibitor. Analysis of PGHS inactivation by AcSHA, coupled with the X-ray crystal structure of the complex of ovine PGHS-1 with AcSHA, confirms that the inhibitor elicits its effects via acetylation of Ser-529 in the cyclooxygenase active site. The crystal structure reveals an intact inhibitor molecule bound in the enzyme's cyclooxygenase active-site channel, hydrogen bonding with Arg-119 of the enzyme. The structure-activity profile of AcSHA can be rationalized in terms of the crystal structure of the enzyme-ligand complex. AcSHA may prove useful as a lead compound to facilitate the development of new acetylating inhibitors.
Subject(s)
Cyclooxygenase Inhibitors/pharmacology , Prostaglandin-Endoperoxide Synthases/metabolism , Salicylamides/pharmacology , Acetylation/drug effects , Animals , Arginine/metabolism , Binding Sites/physiology , COS Cells , Crystallization , Crystallography, X-Ray , Cyclooxygenase 1 , Cyclooxygenase 2 , Cyclooxygenase 2 Inhibitors , Humans , Isoenzymes/antagonists & inhibitors , Isoenzymes/drug effects , Isoenzymes/metabolism , Membrane Proteins , Mutagenesis, Site-Directed , Platelet Aggregation/drug effects , Prostaglandin-Endoperoxide Synthases/chemistry , Prostaglandin-Endoperoxide Synthases/drug effects , Protein Conformation , Serine/metabolismABSTRACT
Several cisplatin analogues of ethylenediamine-derived ligands containing alcohol, carboxylic acid and acetate substituents have been prepared and characterised. Oxidation of some of these square planar platinum(II) complexes using aqueous hydrogen peroxide gave octahedral platinum(IV) complexes, containing trans hydroxo ligands. Acetylation of the hydroxo ligands was achieved by reaction with acetic anhydride, giving complexes which are analogues of the antitumour drug, JM-216. Oxidation of the complex [Pt(H2L4)Cl2], where H2L4 is ethylenediamine-N,N'-diacetic acid, with H2O2 gave the platinum(IV) complex [PtL4Cl2].H2O in which L4 is tetradentate as shown by a crystal and molecular structure. This complex was previously reported to be [Pt(HL4)(OH)Cl2] in which HL4 is tridentate. Several of the complexes were tested for antitumour activity against five human ovarian carcinoma cell lines. IC50 values range from 4.0 microM for cis,trans-PtCl2(OH)2(NH2CH2CH2NHCH2CH2OH) against the CH1 cell line to >25 microM indicating moderate to low activity relative to other platinum complexes.
Subject(s)
Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Organoplatinum Compounds/pharmacology , Antineoplastic Agents/chemical synthesis , Crystallography, X-Ray , Drug Screening Assays, Antitumor , Female , Humans , Ligands , Molecular Structure , Organoplatinum Compounds/chemical synthesis , Organoplatinum Compounds/chemistry , Ovarian Neoplasms , Tumor Cells, CulturedABSTRACT
Reaction of [Ru(Hedta)Cl]- with H2O2 in the presence of arginine, produces NO, in the form of an Ru(II)-(NO+) complex and citrulline which is a remarkably simple model system for the physiological NO synthase reaction.
Subject(s)
Arginine/metabolism , Hydrogen Peroxide/metabolism , Nitric Oxide Synthase/metabolism , Nitric Oxide/metabolism , Ruthenium/metabolism , Citrulline/metabolism , Ruthenium Compounds/metabolismABSTRACT
Stability constants of iron(III), copper(II), nickel(II) and zinc(II) complexes of salicylhydroxamic acid (H2Sha), anthranilic hydroxamic acid (HAha) and benzohydroxamic acid (HBha) have been determined at 25.0 degrees C, I=0.2 mol dm(-3) KCl in aqueous solution. The complex stability order, iron(III) >> copper(II) > nickel(II) approximately = zinc(II) was observed whilst complexes of H2Sha were found to be more stable than those of the other two ligands. In the preparation of ternary metal ion complexes of these ligands and 1,10-phenanthroline (phen) the crystalline complex [Cu(phen)2(Cl)]Cl x H2Sha was obtained and its crystal structure determined. This complex is a model for hydroxamate-peroxidase inhibitor interactions.
Subject(s)
Enzyme Inhibitors/chemistry , Hydroxamic Acids/chemistry , Metals/chemistry , Peroxidases/chemistry , Salicylamides/chemistry , ortho-Aminobenzoates/chemistry , Copper/chemistry , Enzyme Inhibitors/pharmacology , Iron/chemistry , Kinetics , Models, Molecular , Molecular Conformation , Nickel/chemistry , Peroxidases/antagonists & inhibitors , Phenanthrolines/chemistry , Zinc/chemistryABSTRACT
The first solution studies at physiological pH for the formation of metal complexes of taurine, +NH3CH2CH2S03-, one of the most abundant low molecular weight organic compounds in the animal kingdom, are reported. The complexes Cu(Gly-GlyH-1) (1) and [Cu(Gly-AspH-1)] (2) react with taurine to give the ternary complexes [Cu(Gly-GlyH-1)taurine]- (3) (log K=2.95+/-0.03, I=0.2M, T=25.0 degrees C) and [Cu(Gly-AspH-1)taurine]2- (4) (log K=2.68+/-0.02) in which taurine acts as an N-donor ligand, most likely monodentate, without involvement of the sulphonate group in coordination. The results of the pH-metric studies are confirmed by visible and EPR spectrophotometric studies. The taurine complexes are less stable than the analogous complexes of beta-alanine due to the decreased basicity of the amino group in the former ligand, and in the case of the Cu(Gly-GlyH-1) complexes due to involvement of the carboxylate group of beta-alanine in axial coordination.
Subject(s)
Copper/chemistry , Dipeptides/chemistry , Glycine/chemistry , Organometallic Compounds/chemistry , Taurine/chemistry , Animals , Electron Spin Resonance Spectroscopy , Humans , Hydrogen-Ion Concentration , Ligands , Software , Spectrophotometry , Taurine/physiologyABSTRACT
The synthesis and crystal and molecular structures of the platinum(II) complex Pt(HL)Cl where H(2)L is the diacid diamide -[CH(2)N(CH(2)COOH)CH(2)CONH(2)](2), a hydrolytic metabolite of an antitumour active bis(3,5-dioxopiperazin-1-yl)alkane are reported. The complex is square planar and contains HL(-) as a tridentate 2N (amino), O (carboxylate) donor. The metal to ligand bond distances are Pt-Cl 2.287(1) A, Pt-O 2.002 (1) A, Pt-N(trans Cl) 2.014(1) A and Pt-N(trans O) 2.073 A. There is extensive hydrogen bonding, each molecule of Pt(HL)Cl being intermolecularly hydrogen bonded to ten others giving a 3-dimensional network. There is also one intramolecular H-bond.
