In this study, various N-heterocyclic nitro prodrugs (NHN1-16) containing pyrimidine, triazine and piperazine rings were designed and synthesized. The final compounds were identified using FT-IR, ¹H NMR, ¹³C NMR as well as elemental analyses. Enzymatic activities of compounds were conducted by using HPLC analysis to investigate the interaction of substrates with Ssap-NtrB nitroreductase enzyme. MTT assay was performed to evaluate the toxic effect of compounds against Hep3B and PC3 cancer cell lines and healthy HUVEC cell. It was observed that synthesized compounds NHN1-16 exhibited different cytotoxic profiles. Pyrimidine derivative NHN3 and triazine derivative NHN5 can be good drug candidates for prostate cancer with IC₅₀ values of 54.75 µM and 48.9 µM, respectively. Compounds NHN6, NHN10, NHN12, NHN14 and NHN16 were selected as prodrug candidates because of non-toxic properties against three different cell models. The NHN prodrugs and Ssap-NtrB combinations were applied to SRB assay to reveal the prodrug capabilities of these selected compounds. SRB screening results showed that the metabolites of all selected non-toxic compounds showed remarkable cytotoxicity with IC₅₀ values in the range of 1.71–4.72 nM on prostate cancer. Among the tested compounds, especially piperazine derivatives NHN12 and NHN14 showed significant toxic effect with IC₅₀ values of 1.75 nM and 1.79 nM against PC3 cell compared with standart prodrug CB1954 (IC₅₀: 1.71 nM). Novel compounds NHN12 and NHN14 can be considered as promising prodrug candidates for nitroreductase-prodrug based prostate cancer therapy.

在这项研究中，设计并合成了各种含有嘧啶，三嗪和哌嗪环的N杂环硝基前药（NHN1-16）。使用FT-IR，¹ H NMR，¹³ C NMR以及元素分析鉴定最终化合物。通过使用HPLC分析来研究化合物的酶活性，以研究底物与Ssap-NtrB硝基还原酶的相互作用。进行MTT测定以评估化合物对Hep3B和PC3癌细胞系以及健康的HUVEC细胞的毒性作用。观察到合成的化合物NHN1-16表现出不同的细胞毒性特征。嘧啶衍生物NHN3和三嗪衍生物NHN5可以作为前列腺癌的良好候选药物，其IC ₅₀值分别为54.75 µM和48.9 µM。选择化合物NHN6，NHN10，NHN12，NHN14和NHN16作为前药候选药物，因为它们对三种不同的细胞模型均无毒。将NHN前药和Ssap-NtrB组合用于SRB分析，以揭示这些选定化合物的前药能力。SRB筛选结果表明，所有选定的无毒化合物的代谢物均显示出显着的细胞毒性，对前列腺癌的IC ₅₀值为1.71–4.72 nM。在测试的化合物中，尤其是哌嗪衍生物NHN12和NHN14对IC具有明显的毒性作用与标准的前药CB1954相比，针对PC3细胞的₅₀值为1.75 nM和1.79 nM（IC ₅₀：1.71 nM）。新型化合物NHN12和NHN14可被视为基于硝基还原酶-前药的前列腺癌治疗的有前途的候选药物。