title: Mitigation of nitrogen mustard mediated skin injury by a novel indomethacin bifunctional prodrug.
abstract: Nitrogen mustard (NM) is a bifunctional alkylating agent that is highly reactive in the skin causing extensive tissue damage and blistering. In the present studies, a modified cutaneous murine patch model was developed to characterize NM-induced injury and to evaluate the efficacy of an indomethacin pro-drug in mitigating toxicity. NM (20μmol) or vehicle control was applied onto 6mm glass microfiber filters affixed to the shaved dorsal skin of CD-1 mice for 6min. This resulted in absorption of approximately 4μmol of NM. NM caused localized skin damage within 1 d, progressing to an eschar within 2-3 d, followed by wound healing after 4-5 d. NM-induced injury was associated with increases in skin thickness, inflammatory cell infiltration, reduced numbers of sebocytes, basal keratinocyte double stranded DNA breaks, as measured by phospho-histone 2A.X expression, mast cell degranulation and increases in inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Wound healing was characterized by epidermal hyperplasia and marked increases in basal cells expressing proliferating cell nuclear antigen. A novel indomethacin-anticholinergic prodrug (4338) designed to target cyclooxygenases and acetylcholinesterase (AChE), was found to markedly suppress NM toxicity, decreasing wound thickness and eschar formation. The prodrug also inhibited mast cell degranulation, suppressed keratinocyte expression of iNOS and COX-2, as well as markers of epidermal proliferation. These findings indicate that a novel bifunctional pro-drug is effective in limiting NM mediated dermal injury. Moreover, our newly developed cutaneous patch model is a sensitive and reproducible method to assess the mechanism of action of countermeasures.
authors.affiliation: Department of Pharmacology and Toxicology, Rutgers University Ernest Mario School of Pharmacy, Piscataway, NJ, United States.
authors.forename: Gabriella M
authors.initials: GM
authors.lastname: Composto
authors.affiliation: Department of Environmental and Occupational Health, Rutgers University School of Public Health, Piscataway, NJ, United States.
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Laskin
authors.affiliation: Department of Pharmacology and Toxicology, Rutgers University Ernest Mario School of Pharmacy, Piscataway, NJ, United States.
authors.forename: Debra L
authors.initials: DL
authors.lastname: Laskin
authors.affiliation: Department of Pharmacology and Toxicology, Rutgers University Ernest Mario School of Pharmacy, Piscataway, NJ, United States.
authors.forename: Donald R
authors.initials: DR
authors.lastname: Gerecke
authors.affiliation: MRIGlobal, Kansas City, MO, United States.
authors.forename: Robert P
authors.initials: RP
authors.lastname: Casillas
authors.affiliation: Department of Chemistry, Lehigh University, Bethlehem, PA, United States.
authors.forename: Ned D
authors.initials: ND
authors.lastname: Heindel
authors.affiliation: Department of Pharmacology and Toxicology, Rutgers University Ernest Mario School of Pharmacy, Piscataway, NJ, United States.
authors.forename: Laurie B
authors.initials: LB
authors.lastname: Joseph
authors.affiliation: Department of Environmental Health Science, New York Medical College, Valhalla, NY, United States. Electronic address: Diane_Heck@nymc.edu.
authors.forename: Diane E
authors.initials: DE
authors.lastname: Heck
mesh_terms: D000477:Alkylating Agents / Q000633:toxicity; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000737:chemistry / Q000494:pharmacology*; D018680:Cholinergic Antagonists / Q000737:chemistry / Q000494:pharmacology; D051546:Cyclooxygenase 2 / Q000378:metabolism; D016861:Cyclooxygenase Inhibitors / Q000737:chemistry / Q000494:pharmacology; D004249:DNA Damage; D005260:Female; D006657:Histones / Q000378:metabolism; D007150:Immunohistochemistry; D007213:Indomethacin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism; D008466:Mechlorethamine / Q000633:toxicity*; D051379:Mice; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D011355:Prodrugs / Q000737:chemistry / Q000494:pharmacology*; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D012867:Skin / Q000187:drug effects* / Q000293:injuries / Q000473:pathology; D013997:Time Factors; D014945:Wound Healing / Q000187:drug effects
chemical_list: D000477:Alkylating Agents; D000893:Anti-Inflammatory Agents; D018680:Cholinergic Antagonists; D051546:Cyclooxygenase 2; D016861:Cyclooxygenase Inhibitors; D006657:Histones; D007213:Indomethacin; D008466:Mechlorethamine; D052247:Nitric Oxide Synthase Type II; D011355:Prodrugs; D018809:Proliferating Cell Nuclear Antigen; C000620183:4338 compound; C475961:gamma-H2AX protein, mouse
keywords: Anti-cholinergic; Countermeasures; Epidermis; Sulfur mustard; Vesicants