Evaluation of the nitrosamine impurities of ACE inhibitors using computational, in vitro, and in vivo methods demonstrate no genotoxic potential

Abstract Evaluation and mitigation of the potential carcinogenic risks associated with nitrosamines in marketed pharmaceutical products are areas of interest for pharmaceutical companies and health authorities alike. Significant progress has been made to establish acceptable intake (AI) levels for N ‐nitrosamine drug substance‐related impurities (NDSRIs) using SAR, however some compounds require experimental data to support derivation of a recommended AI. Many angiotensin‐converting enzyme inhibitors, identified by the suffix “pril,” have secondary amines that can potentially react to form nitrosamines. Here we consider a structural assessment and metabolism data, coupled with comprehensive in vitro and in vivo (mouse) genotoxicity testing to evaluate this particular class of nitrosamines. N ‐nitroso ramipril and N ‐nitroso quinapril, both of which are predicted to have inhibited nitrosamine bioactivation due to steric hinderance and branching at the α‐position were non‐genotoxic in the in vivo liver comet assay and non‐mutagenic in the in vivo Big Blue® mutation and duplex sequencing assays. Predicted metabolism along with in vitro metabolism data and quantum chemical calculations related to DNA interactions offer a molecular basis for the negative results observed in both in vitro and in vivo testing. These nitrosamines are concluded to be non‐mutagenic and non‐carcinogenic; therefore, they should be controlled according to ICH Q3B guidance. Furthermore, these results for N ‐nitroso ramipril and N ‐nitroso quinapril should be considered when evaluating the appropriate AI and control strategy for other structurally similar “pril” NDSRIs.