title: In vitro exposure of precision-cut lung slices to 2-(4-amino-3-methylphenyl)-5-fluorobenzothiazole lysylamide dihydrochloride (NSC 710305, Phortress) increases inflammatory cytokine content and tissue damage.
abstract: The anticancer drug (2-[4-amino-3-methylphenyl]-5-fluorobenzothiazole lysylamide dihydrochloride) (NSC 710305, Phortress) is a metabolically activated prodrug that causes DNA adduct formation and subsequent toxicity. Preclinically, it was found that hepatic, bone marrow, and pulmonary toxicity presented challenges to developing this drug. An ex vivo precision-cut lung slice (PCLS) model was used to search for concentration dependent effects of NSC 710305 (10, 25, 50, and 100 µM) on cytokine content, protein content, and immuno/histological endpoints. Preparation and culture of PCLS caused an initial spike in proinflammatory cytokine expression and therefore treatment with NSC 710305 was delayed until 48 h after initiating the slice cultures to avoid confounding the response to slicing with any drug response. PCLSs were evaluated after 24, 48, and 72 h exposures to NSC 710305. Reversibility of toxicity due to the 72-h treatment was evaluated after a 24-h recovery period. NSC 710305 caused a concentration-dependent cytokine response, and only the toxicity caused by a 72-h exposure to 25 µM reversed during the 24-h recovery period. Immuno/histological examination and quantitation of tissue protein levels indicated that tissue destruction, ED-1 (activated macrophage) staining, and protein levels were associated with the levels of proinflammatory cytokines in the tissue. In conclusion, the concentration- and time-dependent inflammatory response of PCLS to NSC 710305 preceded relevant tissue damage by a few days. The no-observable adverse effect level (NOAEL) for 24, 48, and 72 h exposures was established as 10 µM NSC 710305.
authors.affiliation: Laboratory of Investigative & Screening Toxicology, LHTP, SAIC-Frederick, Inc., Frederick National Laboratory for Cancer Research, Frederick, Maryland 21702, USA. behrsingh@mail.nih.gov
authors.forename: Holger P
authors.initials: HP
authors.lastname: Behrsing
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Furniss
authors.forename: Myrtle
authors.initials: M
authors.lastname: Davis
authors.forename: Joseph E
authors.initials: JE
authors.lastname: Tomaszewski
authors.forename: Ralph E
authors.initials: RE
authors.lastname: Parchment
mesh_terms: D000818:Animals; D016207:Cytokines / Q000378:metabolism*; D066298:In Vitro Techniques; D018836:Inflammation Mediators / Q000378:metabolism*; D008168:Lung / Q000187:drug effects* / Q000473:pathology; D008239:Lysine / Q000031:analogs & derivatives* / Q000633:toxicity; D008297:Male; D051381:Rats; D011916:Rats, Inbred F344; D013844:Thiazoles / Q000633:toxicity*
chemical_list: C446999:2,6-diaminohexanoic acid (4-(5-fluorobenzothiazol-2-yl)-2-methylphenyl)amide; D016207:Cytokines; D018836:Inflammation Mediators; D013844:Thiazoles; D008239:Lysine