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.

(Z)-2-(3,4-Dichlorophenyl)-3-(1H-Pyrrol-2-yl)Acrylonitrile Exhibits Selective Antitumor Activity in Breast Cancer Cell Lines via the Aryl Hydrocarbon Receptor Pathway

“…Thus, whereas halogenated aryl-hydrocarbons and acrylonitrile compounds have been shown to activate the AhR pathway, the specific steps and mechanisms do differ, underscoring the importance of characterizing the mode of action. Exploring the differential effects of AhR agonists open the way to exploit their subtle differences for the clinical treatment of disease, including management of potential clinical toxicities, which have been described for prodrugs of aminoflavone and 5F-203 (Behrsing et al, 2013; https://dctd.cancer.gov/featuredagents/pdfs/710464aminoflavonetoxabstract.pdf).…”

Applicability of rat precision-cut lung slices in evaluating nanomaterial cytotoxicity, apoptosis, oxidative stress, and inflammation

“…They further indicate that a certain degree of biological variability is to be expected in PCLuS test systems. Recently, it has been suggested to delay exposing PCLuS to test substances until 48 h after preparing the slices to reduce responses caused by the slicing procedure itself (Behrsing et al, 2013). Under the testing conditions of the present study, however, cell viability Table 9 Fulfillment of acceptance criteria (AC) and reproducibility of test results in the protein, WST-1, GSH, caspase-3/-7, and cytokine assays (numbers of NC and PC not meeting the AC and numbers of measurements with CV% N20% and N50% as compared to reported mean intra-and interassay variabilities).…”

Aryl Hydrocarbon Receptor Ligand 5F 203 Induces Oxidative Stress That Triggers DNA Damage in Human Breast Cancer Cells

“…5–7 5F 203 is an AhR agonist 8 that activates the AhR signaling pathway and causes DNA damage, cell cycle arrest, and apoptosis in breast cancer cells that are sensitive to its cytotoxic actions. 9–12 Similar to the mechanism of other AhR agonists, 5F 203 binds to AhR, and the 5F 203-AhR complex translocates from the cytosol to the nucleus.…”