title: Chromatographic separation and biological evaluation of benzimidazole derivative enantiomers as inhibitors of leukotriene biosynthesis.
abstract: For an explicit analysis of the chirality on the effectiveness of a recently identified racemic benzimidazole derivative (BRP7) as inhibitor of leukotriene biosynthesis, we optimized a HPLC-based chiral chromatographic method enabling the quantitative isolation of its enantiomers in sufficient amount to carry out biological investigations. The use of a Lux Amylose-2 column revealed especially profitable to fulfil our task. Indeed, the employment of the amylose-based chiral stationary phase (CSP) in combination with a n-hexane/EtOH/DEA - 99/1/02 (v/v/v) mobile phase allowed getting the enantiomeric peaks fully resolved (α=1.80, RS=2.39). Four consecutive injections repeated at 1-min intervals produced overloaded peaks with a very limited level of isomeric contamination. This procedure allowed the isolation of ca. 20mg of each enantiomer, with enantiomeric excess higher than 99% and 95% for the (S)- and the (R)-isomer, respectively. The enantiomeric elution order was established using synthetic reference compounds of lower enantiomeric excess values. The biological evaluation of the purified individual enantiomers revealed no significant difference in terms of their IC50 values with respect to the previously investigated racemic BRP7: 0.18μM for the (R)-enantiomer (R(2)=0.999) and 0.26μM for the (S)-enantiomer (R(2)=0.986).
authors.affiliation: Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Via del Liceo 1, 06123 Perugia, Italy.
authors.forename: Roccaldo
authors.initials: R
authors.lastname: Sardella
authors.affiliation: Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Gazi University, Etiler, 06330 Yenimahalle, Ankara, Turkey.
authors.forename: Serkan
authors.initials: S
authors.lastname: Levent
authors.affiliation: Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Via del Liceo 1, 06123 Perugia, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Ianni
authors.affiliation: Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Gazi University, Etiler, 06330 Yenimahalle, Ankara, Turkey.
authors.forename: Burcu
authors.initials: B
authors.lastname: Calişkan
authors.affiliation: Institute of Pharmacy, Friedrich-Schiller-University Jena, Philosophenweg 14, D-07743 Jena, Germany.
authors.forename: Jana
authors.initials: J
authors.lastname: Gerstmeier
authors.affiliation: Institute of Pharmacy, Friedrich-Schiller-University Jena, Philosophenweg 14, D-07743 Jena, Germany.
authors.forename: Carlo
authors.initials: C
authors.lastname: Pergola
authors.affiliation: Institute of Pharmacy, Friedrich-Schiller-University Jena, Philosophenweg 14, D-07743 Jena, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Werz
authors.affiliation: Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Gazi University, Etiler, 06330 Yenimahalle, Ankara, Turkey.
authors.forename: Erden
authors.initials: E
authors.lastname: Banoglu
authors.affiliation: Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Via del Liceo 1, 06123 Perugia, Italy. Electronic address: natalini@chimfarm.unipg.it.
authors.forename: Benedetto
authors.initials: B
authors.lastname: Natalini
mesh_terms: D000688:Amylose / Q000737:chemistry; D001562:Benzimidazoles / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004340:Drug Contamination; D006586:Hexanes / Q000737:chemistry; D006801:Humans; D020024:Leukotriene Antagonists / Q000737:chemistry*; D015289:Leukotrienes / Q000378:metabolism*; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism
chemical_list: D001562:Benzimidazoles; D006586:Hexanes; D020024:Leukotriene Antagonists; D015289:Leukotrienes; C026385:n-hexane; D000688:Amylose; C031000:benzimidazole
keywords: 5-Lipoxygenase-activating protein; Benzimidazole; Chromatographic enantioisolation; FLAP; Polysaccharide-based stationary phases