title: Daunorubicin coupled to monoclonal antibodies via a cis-aconitic anhydride linker: biochemical and cytotoxic properties revisited.
abstract: Daunorubicin (DNR) was coupled to monoclonal antibodies (Mab) reactive to breast tumor cells using the acid-labile linking agents cis-asonitic anhydride and two other non acid-labile analogs, glutaric anhydride and citraconic anhydride. The acid derivatives of DNR formed by reaction with the anhydrides were converted to their N-hydroxysuccinimide (NHS) active esters for coupling to MAb. The molar input of drug NHS ester to MAb ranged from 1:1 to 100:1. The resulting MAb-DNR conjugates were purified by gel filtration and analyzed by high performance liquid chromatography. Monomeric conjugates contained 0.2 to 11.0 moles of DNR/mole of MAb. No evidence of cell killing was observed up to a concentration of 10 micrograms/ml DNR bound to MAb, while DNR exhibited 50% killing of the breast tumor cell line MCF-7 at a concentration of 1 microgram/ml.
authors.affiliation: Biotherapeutics, Inc., Franklin, TN 37064.
authors.forename: A
authors.initials: A
authors.lastname: Ahmad
authors.forename: K L
authors.initials: KL
authors.lastname: Law
authors.forename: B
authors.initials: B
authors.lastname: Young
authors.forename: M W
authors.initials: MW
authors.lastname: Telander
authors.forename: J R
authors.initials: JR
authors.lastname: Ogden
authors.forename: K
authors.initials: K
authors.lastname: Leung
mesh_terms: D000156:Aconitic Acid / Q000031:analogs & derivatives; D000911:Antibodies, Monoclonal / Q000302:isolation & purification / Q000494:pharmacology*; D001943:Breast Neoplasms; D002470:Cell Survival / Q000187:drug effects*; D002851:Chromatography, High Pressure Liquid; D003630:Daunorubicin / Q000494:pharmacology*; D006801:Humans; D007202:Indicators and Reagents; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive; D015394:Molecular Structure; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000187:drug effects*
chemical_list: D000911:Antibodies, Monoclonal; D007202:Indicators and Reagents; C064873:aconitic anhydride; D000156:Aconitic Acid; D003630:Daunorubicin