title: Development of an in vitro liquid chromatography-mass spectrometry method to evaluate stereo and chemical stability of new drug candidates employing immobilized artificial membrane column.
abstract: A stopped-flow HPLC method was developed to evaluate configurational and chemical stability of pharmaceutical compounds employing immobilized artificial membranes (IAM) column to simulate conditions that pharmaceutical compounds will meet in vivo. The method was applied to recent developed chiral 5-arylbenzothiadiazine derivatives possessing high positive allosteric modulatory (PAM) activity on AMPA receptor. In particular the stopped-flow HPLC method developed used a chiral column to separate single enantiomer of the compounds that are forced into an IAM column where configurational and chemical stability was evaluated in simulated gastrointestinal fluids (pH 1.2 and 6.8 at 37.5 °C) to simulate in vivo conditions. The results were compared to those obtained by dynamic and off-column methods to evaluate the effects of stationary phases on kinetic constant of enantiomerization and hydrolysis. The results suggested that the phospholipids environment of IAM stationary phases, which mimes biological membrane, greatly influence the hydrolysis process increasing the chemical stability of tested compounds while no influence on enantiomerization kinetic was observed. Therefore it is possible to suppose that 5-arylbenzothiadiazine derivatives should not hydrolysed in vivo while they should rapidly racemized in aqueous solvents. The method could represents a rapid and value tool to predict chemical and configurational stability of new chemical entities to decrease the number of animal studies.
authors.affiliation: Department of Life Sciences, University of Modena & Reggio Emilia, Via Campi 183, 41125 Modena, Italy. Electronic address: giuseppe.cannazza@unimore.it.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Cannazza
authors.affiliation: Department of Life Sciences, University of Modena & Reggio Emilia, Via Campi 183, 41125 Modena, Italy.
authors.forename: Umberto M
authors.initials: UM
authors.lastname: Battisti
authors.affiliation: Department of Biological and Environmental Sciences and Technologies, University of Salento, via Prov.le Lecce-Monteroni, 73100 Lecce, Italy.
authors.forename: Marina M
authors.initials: MM
authors.lastname: Carrozzo
authors.affiliation: Department of Life Sciences, University of Modena & Reggio Emilia, Via Campi 183, 41125 Modena, Italy.
authors.forename: Addolorata S
authors.initials: AS
authors.lastname: Cazzato
authors.affiliation: Department of Life Sciences, University of Modena & Reggio Emilia, Via Campi 183, 41125 Modena, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Braghiroli
authors.affiliation: Department of Life Sciences, University of Modena & Reggio Emilia, Via Campi 183, 41125 Modena, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Parenti
authors.affiliation: Department of Biological and Environmental Sciences and Technologies, University of Salento, via Prov.le Lecce-Monteroni, 73100 Lecce, Italy.
authors.forename: Luigino
authors.initials: L
authors.lastname: Troisi
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods*; D006868:Hydrolysis; D066298:In Vitro Techniques; D007700:Kinetics; D013058:Mass Spectrometry / Q000379:methods*; D008567:Membranes, Artificial; D010743:Phospholipids / Q000737:chemistry; D012997:Solvents; D013237:Stereoisomerism
chemical_list: D008567:Membranes, Artificial; D010743:Phospholipids; D012997:Solvents
keywords: Benzothiadiazines; Chiral OD column; Hydrolysis; IAM column; Racemization