title: Albumin conjugates of the anticancer drug chlorambucil: synthesis, characterization, and in vitro efficacy.
abstract: In our efforts to improve the selectivity and toxicity profile of antitumor agents, four maleimide derivatives of chlorambucil (1-4) were bound to thiolated human serum albumin which differ in the stability of the chemical link between drug and spacer. 1 is an aliphatic maleimide ester derivative of chlorambucil, whereas 2-4 are acetaldehyde, acetophenone, and benzaldehyde carboxylic hydrazone derivatives. HPLC stability studies at pH 5.0 with the related model compounds 5, 7, 8, and 9, in which chlorambucil was substituted by 4-phenylbutyric acid, demonstrated that the carboxylic hydrazone derivatives have acid-sensitive properties; the acid lability of 7 was particular prominent with a half-life of only a few hours. The alkylating activity of albumin-bound chlorambucil was determined with the aid of 4-(4-nitrobenzyl)-pyridine (NBP), demonstrating that on average three equivalents were protein-bound. Evaluation of the cytotoxicity of free chlorambucil and the respective albumin conjugates in the MCF7 mamma carcinoma and MOLT4 leukemia cell line employing a propidium iodide fluorescence assay demonstrated that the conjugate in which chlorambucil was bound to albumin through an ester bond was not as active as chlorambucil. In contrast, the conjugates in which chlorambucil was bound to albumin through carboxylic hydrazone bonds were as or more active than chlorambucil in both cell lines. In particular, the conjugate in which chlorambucil was bound to albumin through an acetaldehyde carboxylic hydrazone bond exhibited IC50 values which were approximately 4-fold (MCF7) to 13-fold (MOLT4) lower than those of chlorambucil. Preliminary toxicity studies in mice showed that this conjugate can be administered at higher doses in comparison to unbound chlorambucil.
authors.affiliation: Tumor Biology Center, Dept. Med. Oncology, Freiburg, Germany.
authors.forename: F
authors.initials: F
authors.lastname: Kratz
authors.forename: U
authors.initials: U
authors.lastname: Beyer
authors.forename: T
authors.initials: T
authors.lastname: Roth
authors.forename: M T
authors.initials: MT
authors.lastname: Schütte
authors.forename: A
authors.initials: A
authors.lastname: Unold
authors.forename: H H
authors.initials: HH
authors.lastname: Fiebig
authors.forename: C
authors.initials: C
authors.lastname: Unger
mesh_terms: D000818:Animals; D018906:Antineoplastic Agents, Alkylating / Q000138:chemical synthesis*; D002699:Chlorambucil / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D004355:Drug Stability; D005260:Female; D006801:Humans; D051379:Mice; D012709:Serum Albumin
chemical_list: D018906:Antineoplastic Agents, Alkylating; D012709:Serum Albumin; D002699:Chlorambucil