title: An HPLC/mass spectrometry platform for the development of multimodality contrast agents and targeted therapeutics: prostate-specific membrane antigen small molecule derivatives.
abstract: The production of disease-targeted agents requires the covalent conjugation of a targeting molecule with a contrast agent or therapeutic, followed by purification of the product to homogeneity. Typical targeting molecules, such as small molecules and peptides, often have high charge-to-mass ratios and/or hydrophobicity. Contrast agents and therapeutics themselves are also diverse, and include lanthanide chelates for MRI, (99m)Tc chelates for SPECT, (90)Y chelates for radiotherapy, (18)F derivatives for PET, and heptamethine indocyanines for near-infrared fluorescent optical imaging. We have constructed a general-purpose HPLC/mass spectrometry platform capable of purifying virtually any targeted agent for any modality. The analytical sub-system is composed of a single dual-head pump that directs mobile phase to either a hot cell for the purification of radioactive agents or to an ES-TOF MS for the purification of nonradioactive agents. Nonradioactive agents are also monitored during purification by ELSD, absorbance and fluorescence. The preparative sub-system is composed of columns and procedures that permit rapid scaling from the analytical system. To demonstrate the platform's utility, we describe the preparation of five small molecule derivatives specific for prostate-specific membrane antigen (PSMA): a gadolinium derivative for MRI, indium, rhenium and technetium derivatives for SPECT, and an yttrium derivative for radiotherapy. All five compounds are derived from a highly anionic targeting ligand engineered to have a single nucleophile for N-hydroxysuccinimide-based conjugation. We also describe optimized column/mobile phase combinations and mass spectrometry settings for each class of agent, and discuss strategies for purifying molecules with extreme charge and/or hydrophobicity. Taken together, our study should expedite the development of disease-targeted, multimodality diagnostic and therapeutic agents.
authors.affiliation: Division of Hematology/Oncology, Beth Israel Deaconess Medical Center, Boston, MA 02215, USA.
authors.forename: Valerie
authors.initials: V
authors.lastname: Humblet
authors.forename: Preeti
authors.initials: P
authors.lastname: Misra
authors.forename: John V
authors.initials: JV
authors.lastname: Frangioni
mesh_terms: D000954:Antigens, Surface / Q000737:chemistry* / Q000302:isolation & purification; D045744:Cell Line, Tumor; D002470:Cell Survival; D002851:Chromatography, High Pressure Liquid; D003287:Contrast Media / Q000138:chemical synthesis*; D005682:Gadolinium / Q000737:chemistry; D043425:Glutamate Carboxypeptidase II / Q000737:chemistry* / Q000302:isolation & purification; D017261:Glycosylphosphatidylinositols / Q000737:chemistry; D006573:Heterocyclic Compounds, 1-Ring / Q000737:chemistry; D006801:Humans; D007811:Lanthanum / Q000737:chemistry; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D019275:Radiopharmaceuticals / Q000138:chemical synthesis* / Q000737:chemistry; D015018:Ytterbium / Q000737:chemistry
chemical_list: D000954:Antigens, Surface; D003287:Contrast Media; D017261:Glycosylphosphatidylinositols; D006573:Heterocyclic Compounds, 1-Ring; D019275:Radiopharmaceuticals; C071349:1,4,7,10-tetraazacyclododecane- 1,4,7,10-tetraacetic acid; D007811:Lanthanum; D005682:Gadolinium; C473848:FOLH1 protein, human; D043425:Glutamate Carboxypeptidase II; D015018:Ytterbium