title: Generation of polyclonal catalytic antibodies against cocaine using transition state analogs of cocaine conjugated to diphtheria toxoid.
abstract: Six novel transition state analogs (TSAs) of cocaine (10-14 and 17) and one non-cocaine, p-aminophenylphosphonyl ester of cyclohexanol (19), were synthesized and characterized by 1H- and 13C-NMR and FAB-MS. (1R)-ecgonine methyl ester or cyclohexanol were subjected to phenylphosphonylation in the presence of dicyclohexyl carbodiimde (DCC) and 4-N,N-dimethyl aminopyridine (4-DMAP). TSA-IV (10), however, was synthesized from norcocaine which was protected with dibromoethane to yield 4 before acid hydrolysis, esterification and phenylphosphonylation were carried out. TSA-III (11) TSA-I (12) and (19), using various length spacer arms, were coupled with the immunogenic protein, diphtheria toxoid (DT). The TSAs coupled with DT were used to immunize mice and after appropriate boosts their sera were tested for the presence and titer of anti-TSA polyclonal antibodies using ELISA. Preliminary results show that the mice immunized with these TSAs produced high titers of polyclonal catalytic antibodies, except for (19), with the ability to hydrolyze the substrate 125I-4'-iodococaine in an in vitro assay, even in the presence of noncatalytic anti-TSA antibodies.
authors.affiliation: Department of Medicinal Chemistry and Pharmaceutics, College of Pharmacy, University of Oklahoma Health Sciences Center, Oklahoma City 73117, USA.
authors.forename: G P
authors.initials: GP
authors.lastname: Basmadjian
authors.forename: S
authors.initials: S
authors.lastname: Singh
authors.forename: B
authors.initials: B
authors.lastname: Sastrodjojo
authors.forename: B T
authors.initials: BT
authors.lastname: Smith
authors.forename: K S
authors.initials: KS
authors.lastname: Avor
authors.forename: F
authors.initials: F
authors.lastname: Chang
authors.forename: S L
authors.initials: SL
authors.lastname: Mills
authors.forename: T W
authors.initials: TW
authors.lastname: Seale
mesh_terms: D015761:4-Aminopyridine / Q000031:analogs & derivatives / Q000737:chemistry; D000818:Animals; D000906:Antibodies / Q000494:pharmacology*; D000918:Antibody Specificity; D002384:Catalysis; D003042:Cocaine / Q000031:analogs & derivatives* / Q000276:immunology* / Q000378:metabolism; D004024:Dicyclohexylcarbodiimide / Q000737:chemistry; D004168:Diphtheria Toxoid; D004797:Enzyme-Linked Immunosorbent Assay; D004951:Esterification; D006868:Hydrolysis; D066298:In Vitro Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D010766:Phosphorylation; D013329:Structure-Activity Relationship
chemical_list: D000906:Antibodies; D004168:Diphtheria Toxoid; D004024:Dicyclohexylcarbodiimide; D015761:4-Aminopyridine; D003042:Cocaine; C003885:4-dimethylaminopyridine