Ames concordance with the in vivo transgenic rodent (TGR) gene mutation assay for NDSRIs and relative in vivo TGR potency with nitrosamines with robust dose-response carcinogenicity data.

Nitrosamines (NAs) are a diverse class of mutagenic impurities encompassing both small molecules and structurally complex drug-related NAs, referred to as nitrosamine drug substance-related impurities (NDSRIs). NAs display a broad range of carcinogenic potential, from high carcinogenic potency to being weak or even non-carcinogenic. In vitro Ames tests, conducted with both rat and hamster liver-induced S9, and in vivo transgenic rodent (TGR) mutation assays have been used by pharmaceutical sponsors for hazard identification of NDSRIs. A comparative analysis of Ames tests and TGR results for 33 NDSRIs was performed and revealed an accuracy of 79 % between the overall mutagenic calls in the two assays. For NDSRIs with positive TGR results, mutagenic potency estimates were calculated and compared to NAs with robust carcinogenicity and TGR dose-response data. Results from these NAs demonstrated a strong correlation between carcinogenic potency (TD50) and TGR mutagenic potency (BMDL50) (r2 = 0.95), which supports the use of TGR data for both hazard identification and acceptable intake (AI) determination. By integrating quantitative risk assessment tools with TGR assays, this work contributes to a more robust framework for evaluating NA-associated risks.