Design Ang Synthesis Of Phosphonates As Hapten For Genetating Catalytia Antibodies That Catalyzing Enantioselective Hydrolysis Of Ibuprofen Methyl Ester

Ibuprofen, (R,S)-2-(4-isobutylphenyl) propionic acid, is a widely used nonsteroidal anti-inflammatory drug which belongs to the family of 2-arylpropionic acid derivatives. The pharmaceutical activity of 2-arylpropionic acid derivatives is often dramatically dependent on the chirality of these compounds. In the case of ibuprofen, because of the asymmetric carbon in the second position, the S-(+)-enantiomer is known to be about 100 times more active than its R-(-)-enantiomer. Chemical synthesis of the S-isomer involving diastereomeric crystallization from a racemic acid mixture has been reported. Considerable efforts have been made in improving the methods of its asymmetric synthesis. A resolution of racemic ibuprofen has been achieved by enzyme-catalyzed or microbe cells enantioselective hydrolysis of the corresponding racemic esters, amides and nitriles.Phosphonates are recognized as effective transition-state analogue inhibitors for a variety of enzymes including a number of proteases and esterase. Therein, 1-arylethylphosphonates are of interest as potential biologically active compounds, primarily as phosphorus analogs of 2-arylpropionic acids that are known as nonsteroid antiphlogistic and analgetic drugs, such as ibuprofen. It was demonstrated that 1-arylethylphosphonates caused formation in animal organism of antibodies possessing properties of catalysts of stereospecific hydrolysis.In this paper, we describe a new biocatalyst, catalytic antibodies, that produced ibuprofen from their corresponding racemic ester in enantiomerically pure (S)-configuration. The biocatalyzed hydrolysis reaction can be carried out with antibodies to achieve high enantioselectivity. To elicit catalytic antibodies for the ibuprofen methyl ester hydrolysis, the tetrahedron structure compound contained phosphorus was designed and synthesized as hapten which is mimic of TSA, based on the transition state theory. The hapten was covalently attached to bovine serum albumin (BSA) to be used as immunogens. Catalytic antibodies were prepared from mice and sequential purification by ammonium sulfate precipitation and affinity chromatography.Then, Kinetic analysis of the catalytic antibody-catalyzed reaction was found to be possible in a reverse micelle reaction system based on sodium bis (2-ethylhexyl) sodium sulfosuccinate (AOT) in isooctane. Kinetic studies showed that hydrolysis in the microemulsion system follows Michaelis-Menten kinetics.