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Selection of the Most Suitable Source Analogue for 2,3-Benzofluorene Using Read-Across Methodology to Support Derivation of Human Health Toxicity Values

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  Background and Purpose: Chemical hazard assessments derive human health toxicity values for use in the Superfund Program. Toxicity values are derived from a review of the relevant scientific literature using U.S. EPA methods, sources of data, and guidance for value derivation. In some instances, no useful human or animal toxicity data are available for the chemical under evaluation and read-across assessments are attempted. 2,3-Benzofluorene is a polycyclic aromatic hydrocarbon (PAH). Although not produced commercially, 2,3-benzofluorene is formed during the incomplete combustion or pyrolysis of organic matter such as wood and fuel. PAHs, including 2,3-benzofluorene, are also generated as byproducts of industrial processes, such as coal and petroleum refining, and the production of iron, steel, and aluminum. The chemical hazard assessment of 2,3-benzofluorene evaluated the feasibility of deriving provisional oral and inhalation toxicity values.   Methods: Literature searches were conducted to identify studies relevant to the derivation of provisional toxicity values for 2,3-benzofluorene. Repeated-dose, short-term, subchronic, chronic, and reproductive/ developmental toxicity studies were considered relevant study designs to determine if data were suitable for derivation of provisional toxicity values using traditional methods. Because adequate data were not located, an alternative analogue approach was used to identify candidate analogues that could be used to derive screening-level oral or inhalation toxicity values. Structural analogues were initially identified using automated screening tools (e.g., ChemID Plus, CompTox Chemicals Dashboard, OECD Toolbox) and then refined by an experienced chemist based on known or expected structure-toxicity relationships, reactivity, and metabolic pathways. Metabolic analogues (metabolites and metabolic precursors) were identified from metabolism simulators and targeted literature searches. Toxicity/mechanistic/mode-of-action (MOA) analogues were identified from in vivo toxicity data and in vitro mechanistic data (e.g., bioactivity data) for 2,3-benzofluorene. The availability of toxicity values from authoritative sources was determined for the candidate analogues. Candidate analogues with available toxicity values were evaluated across three similarity domains (structural/physicochemical, metabolic/toxicokinetic, and toxicodynamic/MOA) to determine the most suitable source analogue for 2,3-benzofluorene.   Conclusions: Fluorene was selected as the source analogue for the derivation of screening oral toxicity values for 2,3¿benzoflourene. The adoption of oral points-of-departure (PODs) for fluorene to support the development of screening p-RfDs for 2,3-benzofluorene is reasonably expected to be protective of effects resulting from oral exposure to 2,3-benzofluorene.    

Impact/Purpose

Results: The oral toxicity database for 2,3¿benzofluorene was limited to a 5¿day gavage experiment in rats designed as an in vivo transcriptomic study with limited toxicological endpoints and a 14¿day study in mice that only analyzed DNA adduct formation in the lung. No studies were located regarding toxicity of 2,3¿benzofluorene to humans or animals via inhalation exposure. Due to the lack of suitable toxicity data from repeated-dose studies, oral and inhalation toxicity values could not be derived directly. Instead, an alternative analogue approach was used to determine if screening toxicity values could be derived. This approach yielded 10 structural analogues, 27 metabolic analogues, and 1 mechanistic analogue. None of the candidate analogues had inhalation toxicity values; therefore, screening inhalation toxicity values could not be derived. Three of the analogues had oral toxicity values (fluorene, fluoranthene, and acenaphthene) and were considered further. With respect to structural properties, fluorene is most similar to 2,3-benzofluorene (both compounds have one open benzylic position and one bay-like region). Based on physicochemical properties, all candidate analogues are reasonably similar to 2,3¿benzofluorene. Predicted structural alerts for the target chemical and candidate analogues suggest that fluorene is most similar to 2,3-benzofluorene, with several shared alerts that were not common to both fluoranthene and acenaphthene (e.g., hepatotoxicity based on diclofenac alert, renal toxicity alerts based on  2-amino-4,5 diphenyl thiazole, anthraquinone, carbamazepine and propranolol or ticlopidine, etc.) There are apparent differences in the metabolism of the candidate analogues based on the presence and number of open benzylic positions (metabolites for acenaphthene, but not 2,3-benzofluorene or the other candidate analogues, include carboxylic acids). Of the candidate analogues, only fluoranthene has been shown to be metabolized to form reactive epoxide intermediates. Epoxide metabolites involving the bay¿like region are also predicted for 2,3¿benzofluorene; however, unlike PAHs with a true bay region, these epoxides do not have a diol group and are not found in a position analogous to the metabolites for PAHs with a true bay region (which form dihydrodiol epoxides as ultimate carcinogenic metabolites). Limited toxicity data were available for 2,3-benzofluorene. A 5¿day oral toxicity study of 2,3¿benzofluorene in rats, designed primarily as an in vivo transcriptomic study with the assessment of some toxicological endpoints, identified effects on red blood cells (RBCs), liver, and thyroid hormone homeostasis. Available toxicity data for the candidate analogues identified effects on RBCs, liver, and kidney following subchronic oral exposure to fluoranthene and fluorene, and liver effects for acenaphthene. Alerts were indicated by the OECD QSAR Toolbox for hepatic and renal toxicity for 2,3¿benzofluorene and all candidate analogues. Based on a weight-of-evidence (WOE) evaluation for the three categories of evidence, fluorene was considered the most suitable source analogue for 2,3-benzofluorene. Like the target chemical, fluorene has one open benzylic position and one bay¿like region, and both chemicals share similar structural alert profiles. Limited data demonstrate that liver and RBC effects are observed following short-term oral exposure to 2,3¿benzofluorene in rats. Critical effects of fluorene that were selected as the basis for its oral toxicity values from authoritative sources were increased relative liver weight and decreased RBC count, packed cell volume, and hemoglobin.

Citation

Mezencev, R., Luci Lizarraga, Jeff Dean, J. Zhao, K. Salinas, M. Kawa, L. Morlacci, J. Rhoades, S. Nieman, M. Odin, T. Shannon, JonathanPhillip Kaiser, AND Alli Flynn. Selection of the Most Suitable Source Analogue for 2,3-Benzofluorene Using Read-Across Methodology to Support Derivation of Human Health Toxicity Values. Society of Toxicology, Orlando, FL, March 16 - 20, 2025.

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  • BENZOFLUORENE SOT 2025 POSTER_CLEARANCE.PPTX
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Last updated on March 17, 2025
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