Elucidation of the enantioselective recognition mechanism of a penicillin G acylase-based chiral stationary phase towards a series of 2-aryloxy-2-arylacetic acids.

A series of structurally related 2-aryloxy-2-arylacetic acids (1-3, 5-16) together with a thioisostere derivative (4) have been synthesized and characterized by GC-MS and 1H NMR. The designed compounds were analyzed on a Penicillin G Acylase chiral stationary phase (PGA-CSP) and the influence of the structure variations on retention and enantioselectivity was investigated. The chromatographic study includes the direct separation of the enantiomers of the synthesized compounds and the determination of the elution order of selected racemic mixtures. 10 out of 16 racemates were separated; high chromatographic enantioseparation factors (alpha > 2) were achieved for some compounds. For the enantiomers of four compounds whose absolute configuration was known (1, 3, 12, 16), the elution order was R:S with the exception of 2-(4-chloro-phenoxy)phenylacetic acid (1), for which the elution order was reversed. Preliminary molecular modeling studies suggest that both polar and charge-transfer interactions as well as steric effects play an important role in determining the retention factors and the enantioselectivities observed.

[1]  G. Massolini,et al.  Penicillin G acylase-based stationary phases: analytical applications. , 2004, Journal of pharmaceutical and biomedical analysis.

[2]  G. Massolini,et al.  Evaluation of a monolithic epoxy silica support for penicillin G acylase immobilization. , 2004, Journal of chromatography. A.

[3]  A. De Luca,et al.  Structural requisites of 2‐(p‐chlorophenoxy)propionic acid analogues for activity on native rat skeletal muscle chloride conductance and on heterologously expressed CLC‐1 , 2003, British journal of pharmacology.

[4]  G. Massolini,et al.  Enantioselective hydrolysis of some 2-aryloxyalkanoic acid methyl esters and isosteric analogues using a penicillin G acylase-based HPLC monolithic silica column. , 2003, Analytical chemistry.

[5]  David Weiner,et al.  An Enzyme Library Approach to Biocatalysis: Development of Nitrilases for Enantioselective Production of Carboxylic Acid Derivatives , 2002 .

[6]  R. Fernández-Lafuente,et al.  Influence of the enzyme derivative preparation and substrate structure on the enantioselectivity of penicillin G acylase , 2002 .

[7]  G. Massolini,et al.  Evaluation of a penicillin G acylase-based chiral stationary phase towards a series of 2-aryloxyalkanoic acids, isosteric analogs and 2-arylpropionic acids. , 2002, Journal of chromatography. A.

[8]  K. Wilson,et al.  Crystal structures of penicillin acylase enzyme-substrate complexes: structural insights into the catalytic mechanism. , 2001, Journal of molecular biology.

[9]  G. Massolini,et al.  Immobilized penicillin G acylase as reactor and chiral selector in liquid chromatography. , 2001, Journal of chromatography. A.

[10]  A. De Luca,et al.  Carboxylic acids and skeletal muscle chloride channel conductance: effects on the biological activity induced by the introduction of methyl groups on the aromatic ring of chiral alpha-(4-chloro-phenoxy)alkanoic acids. , 2001, Farmaco.

[11]  J. Haginaka.  Protein-based chiral stationary phases for high-performance liquid chromatography enantioseparations. , 2001, Journal of chromatography. A.

[12]  A. De Luca,et al.  Pharmacological characterization of chloride channels belonging to the ClC family by the use of chiral clofibric acid derivatives. , 2000, Molecular pharmacology.

[13]  R E Hubbard,et al.  Ligand-induced conformational change in penicillin acylase. , 1998, Journal of molecular biology.

[14]  David S. Goodsell,et al.  Automated docking using a Lamarckian genetic algorithm and an empirical binding free energy function , 1998, J. Comput. Chem..

[15]  S. Collina,et al.  Chiral resolution, configurational study and pharmacological profile of 2-phenoxypropionic acids. , 1997, Farmaco.

[16]  D. Noonan,et al.  Structural Requirements of Chiral Clofibric Acid Analogs for Activation of the Rat Peroxisome Proliferator‐activated Receptor α , 1996, Annals of the New York Academy of Sciences.

[17]  M. V. Savchenko,et al.  Enantioselective Penicillin Acylase—catalyzed Reactions , 1996 .

[18]  J M Thornton,et al.  LIGPLOT: a program to generate schematic diagrams of protein-ligand interactions. , 1995, Protein engineering.

[19]  J. Siegel,et al.  Interaction between stacked aryl groups in 1,8-diarylnaphthalenes: Dominance of polar/π over charge-transfer effects , 1995 .

[20]  C. Hunter.  Arene—Arene Interactions: Electrostatic or Charge Transfer? , 1993 .

[21]  F. Diederich,et al.  Cyclophane-arene inclusion complexation in protic solvents: solvent effects versus electron donor-acceptor interactions , 1991 .

[22]  D. Witiak,et al.  In vivo and in vitro peroxisome proliferation properties of selected clofibrate analogues in the rat. Structure-activity relationships. , 1990, Biochemical pharmacology.

[23]  F. Loiodice,et al.  Stereospecificity of the chloride ion channel: the action of chiral clofibric acid analogues. , 1987, Journal of medicinal chemistry.

[24]  D. Witiak,et al.  Dissociation of hypolipidemic and antiplatelet actions from adverse myotonic effects of clofibric acid related enantiomers. , 1987, Journal of medicinal chemistry.

[25]  O. Mitsunobu.  The Use of Diethyl Azodicarboxylate and Triphenylphosphine in Synthesis and Transformation of Natural Products , 1981 .

[26]  M. Arroyo,et al.  Biotechnological applications of penicillin acylases: state-of-the-art , 2002, Applied Microbiology and Biotechnology.

[27]  D. Noonan,et al.  Stereoselective effects of chiral clofibric acid analogs on rat peroxisome proliferator-activated receptor ? (rPPAR?) activation and peroxisomal fatty acid ?-oxidation , 1997 .

[28]  H. Waldmann,et al.  NEW ENZYMATIC METHODS FOR THE SELECTIVE FUNCTIONALIZATION OF CARBOHYDRATE-DERIVATIVES , 1992 .

[29]  H. Waldmann.  The use of penicillin acylase for selective N-terminal deprotection in peptide synthesis , 1988 .