Prodrugs for Nitroreductase Based Cancer Therapy-5: Development of Trinitroaniline Prodrugs/Ssap-NtrB Combinations for Liver Cancer Using Intracellular and Extracellular Conditions

Abstract

In this study, a series of trinitroaniline derivatives (TNA1-8) were synthesized and investigated as potential antitumor agents for enzyme-prodrug therapy. Enzymatic efficiency of Ssap-NtrB on prodrug candidates was determined with HPLC analysis and kinetic studies. The anti-proliferative properties of compounds were determined against four cancer cell lines (Hep3B, PC3, HT-29, and Saos-2) and a healthy cell line (HUVEC) via MTT assay. Intracellular and extracellular prodrug-enzyme treatments were carried out on Hep3B cells. Herewith, the expression of the Ssap-NtrB gene was confirmed with this study. IC50 values of piperidine and 1,3-cyclohexyl derivatives (TNA4 and TNA7) were identified as 1.724 nM and 1.640 nM at extracellular conditions. In intracellular conditions, it was determined as 0.293 μM and 0.393 μM, respectively. In summary, 2,4,6-trinitroaniline derivatives, especially compounds TNA4 and TNA7 might be used as prodrug candidates along with Ssap-NtrB for hepatocellular carcinoma therapy and the development of new prodrugs at cancer treatments.