title: Development of easy-to-use reverse-phase liquid chromatographic methods for determining PRE-084, RC-33 and RC-34 in biological matrices. The first step for in vivo analysis of sigma1 receptor agonists.
abstract: Over the years there has been a growing interest in the therapeutic potential for central nervous system pathologies of sigma receptor modulators. The widely studied PRE-084 and our compounds RC-33 and RC-34 are very potent and selective sigma 1 receptor agonists that could represent promising drug candidates for Amyotrophic Lateral Sclerosis (ALS). Herein, we develop and validate robust and easy-to-use reverse-phase chromatographic methods suitable for detecting and quantifying PRE-084, RC-33 and RC-34 in mouse blood, brain and spinal cord. An HPLC/UV/ESI-MS system was employed for analyzing PRE-084 and an HPLC/UV-PDA system for determining RC-33 and RC-34. Chromatographic separations were achieved on Waters Symmetry RP18 column (150 × 3.9 mm, 5 µm), eluting with water and acetonitrile (both containing 0.1% formic acid) in gradient conditions. The recovery of PRE-084, RC-33 and RC-34 was >95% in all the considered matrices. Their limits of quantitation and detection were also determined. Validation proved the methods be suitable for separating tested compounds from endogenous interferences, being characterized by good sensitivity, linearity, precision and accuracy. A preliminary central nervous system distribution study showed a high distribution of RC-33 in brain and spinal cord, with concentration values well above the determined limit of quantitation. The proposed methods will be used in future preclinical investigations.
authors.affiliation: Department of Drug Sciences, Medicinal Chemistry and Pharmaceutical Technology section, University of Pavia, Viale Taramelli 12, 27100, Pavia, Italy.
authors.forename: Annamaria
authors.initials: A
authors.lastname: Marra
authors.affiliation: Department of Drug Sciences, Medicinal Chemistry and Pharmaceutical Technology section, University of Pavia, Viale Taramelli 12, 27100, Pavia, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Rossi
authors.affiliation: Department of Drug Sciences, Medicinal Chemistry and Pharmaceutical Technology section, University of Pavia, Viale Taramelli 12, 27100, Pavia, Italy.
authors.forename: Lauretta
authors.initials: L
authors.lastname: Maggi
authors.affiliation: Centro Grandi Strumenti, University of Pavia, Via bassi 21, 27100, Pavia, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Corana
authors.affiliation: Centro Grandi Strumenti, University of Pavia, Via bassi 21, 27100, Pavia, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Mannucci
authors.affiliation: Department of Biology and Biotechnology 'L. Spallanzani', Laboratory of Cellular and Molecular Neuropharmacology, University of Pavia, Via Ferrata 9, 27100, Pavia, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Peviani
authors.affiliation: Department of Biology and Biotechnology 'L. Spallanzani', Laboratory of Cellular and Molecular Neuropharmacology, University of Pavia, Via Ferrata 9, 27100, Pavia, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Curti
authors.affiliation: Department of Drug Sciences, Medicinal Chemistry and Pharmaceutical Technology section, University of Pavia, Viale Taramelli 12, 27100, Pavia, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Collina
mesh_terms: D000818:Animals; D001713:Biphenyl Compounds / Q000097:blood* / Q000493:pharmacokinetics*; D001921:Brain / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D051379:Mice; D009025:Morpholines / Q000097:blood / Q000493:pharmacokinetics*; D010880:Piperidines / Q000097:blood* / Q000493:pharmacokinetics*; D017480:Receptors, sigma / Q000819:agonists*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D013116:Spinal Cord / Q000378:metabolism; D000097605:Sigma-1 Receptor
chemical_list: D001713:Biphenyl Compounds; D009025:Morpholines; D010880:Piperidines; D017480:Receptors, sigma; D000097605:Sigma-1 Receptor; C584299:1-(3-biphenyl-4-ylbutyl)piperidine; C000612046:RC-34 compound; C071738:2-(4-morpholino)ethyl-1-phenylcyclohexane-1-carboxylate
keywords: HPLC/UV-PDA; HPLC/UV/ESI-MS; biological matrices; method validation; sigma receptor agonists