title: Significant differences in biological parameters between prodrugs cleavable by carboxypeptidase G2 that generate 3,5-difluoro-phenol and -aniline nitrogen mustards in gene-directed enzyme prodrug therapy systems.
abstract: Nine new nitrogen mustard compounds derived from 2,6-difluoro-4-hydroxy- (3a-e) and 2,6-difluoro-4-amino- (4a-d) aniline were synthesized as potential prodrugs. They were designed to be activated to their corresponding 3,5-difluorophenol and -aniline (4)-nitrogen mustards by the enzyme carboxypeptidase G2 (CPG2) in gene-directed enzyme prodrug therapy (GDEPT) models. The compounds were tested for cytotoxicity in the MDA MB-361 breast adenocarcinoma. The cell line was engineered to express stably either CPG2 tethered to the cell surface stCPG2-(Q)3 or beta-galactosidase (beta-Gal) as control. The cytotoxicity differentials were calculated between CPG 2-expressing and -nonexpressing cells and yielded different results for the two series of prodrugs despite their structural similarities. While the phenol compounds are ineffective as prodrugs, their aniline counterparts exhibit outstanding activity in the tumor cell lines expressing CPG2. [3,5-Difluoro-4-[bis(2-chloroethyl)amino]phenyl]carbamoyl-l-glutamic acid gave a differential of >227 in MDA MB361 cells as compared with 19 exhibited by 4-[(2-chloroethyl)(2-mesyloxyethyl)amino]benzoyl-l-glutamic acid, 1a, which has been in clinical trials.
authors.affiliation: Cancer Research-UK Centre for Cancer Therapeutics at the Institute of Cancer Research, 15 Cotswold Road, Sutton, Surrey, SM2 5NG, UK.
authors.forename: I
authors.initials: I
authors.lastname: Niculescu-Duvaz
authors.forename: I
authors.initials: I
authors.lastname: Scanlon
authors.forename: D
authors.initials: D
authors.lastname: Niculescu-Duvaz
authors.forename: F
authors.initials: F
authors.lastname: Friedlos
authors.forename: J
authors.initials: J
authors.lastname: Martin
authors.forename: R
authors.initials: R
authors.lastname: Marais
authors.forename: C J
authors.initials: CJ
authors.lastname: Springer
mesh_terms: D000816:Aniline Mustard / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D000970:Antineoplastic Agents / Q000138:chemical synthesis / Q000378:metabolism* / Q000494:pharmacology; D001555:Benzene Derivatives / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology; D045744:Cell Line, Tumor; D004354:Drug Screening Assays, Antitumor; D005260:Female; D015316:Genetic Therapy; D018698:Glutamic Acid / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000378:metabolism* / Q000494:pharmacology; D006207:Half-Life; D006801:Humans; D051379:Mice; D009368:Neoplasm Transplantation; D009588:Nitrogen Mustard Compounds / Q000138:chemical synthesis / Q000378:metabolism* / Q000494:pharmacology; D011355:Prodrugs / Q000138:chemical synthesis / Q000378:metabolism* / Q000494:pharmacology; D013329:Structure-Activity Relationship; D014183:Transplantation, Heterologous; D011623:gamma-Glutamyl Hydrolase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: C486794:(3,5-difluoro-4-(bis(2-chloroethyl)amino)phenyl)carbamoyl-glutamic acid; D000970:Antineoplastic Agents; D001555:Benzene Derivatives; D009588:Nitrogen Mustard Compounds; D011355:Prodrugs; D018698:Glutamic Acid; D000816:Aniline Mustard; D011623:gamma-Glutamyl Hydrolase