Acidic hydrolytic stability of the Lifitegrast: Degradation, Method development, Validation, Synthesis, LC-MS/NMR Characterization, In silico and In vitro cytotoxicity

In addition to the multiple vision problems, the toxicity of the degradation products of ophthalmic medications on eye health is today’s concern. Hence, degradation studies of ophthalmic drug and toxicity establishment of the degradation products have been considered. The manuscript includes simple, mass-compatible HPLC methodology and comprehensive protocol for forced degradation studies of the lifitegrast (LFT). Two degradants, (S)-2-(5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methylsulfonyl)phenyl) propanoic acid, (DP1) and benzofuran-6-carboxylicacid (DP2) were formed under acidic stress conditions. An accurate, robust HPLC method has been validated for routine quality control analysis of LFT. Theoretical quantum chemical calculations of reaction enthalpy were studied to explain the formation of DP1 and DP2. A synthesis scheme for DP1 was employed, and characterised using LC-ESI-MS, NMR techniques. The characterization of DP2 was conducted using various analytical advancements. By employing Swiss ADME software, the details of DPs concerning various physicochemical characteristics, and skin permeation capabilities were predicted. A DEREK prediction has revealed the meta-structures in the test molecules which have adverse toxicological properties. Human corneal epithelial (HCE) cell viability assessment upon treatment with LFT DPs revealed that the compound DP2 showed significant cytotoxicity at lower concentration (8 µg/mL) whereas DP1 showed significant toxicity only at higher concentrations (1280 µg/mL).