Biological Activity and Physicochemical Properties of Dipeptidyl Nitrile Derivatives Against Pancreatic Ductal Adenocarcinoma Cells

Autor(es)

Quilles Jr, Jose C. ; Bernardi, Murillo D. L. ^{[1, 2]} ; Batista, Pedro H. J. ^[1] ; Silva, Samelyn C. M. ^[1] ; Rocha, Camila M. R. ^{[1, 2]} ; Montanari, Carlos A. ^[1] ; Leitao, Andrei ^{[1, 2]}

Afiliação do(s) autor(es)

^[1] Quilles Jr, Jr., Jose C., Univ Sao Paulo, Sao Carlos Inst Chem IQSC, Med Chem Grp NEQUIMED, Av Trabalhador Sao Carlense 400, Sao Carlos, SP - Brazil

^[2] Univ Sao Paulo, PPGIB, Av Trabalhador Sao Carlense 400, Sao Carlos, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: ANTI-CANCER AGENTS IN MEDICINAL CHEMISTRY; v. 19, n. 1, p. 112-120, 2019.
• Citações Web of Science: 2

Resumo

Background: Pancreatic cancer is one of the most aggressive types with high mortality in patients. Therefore, studies to discover new drugs based on cellular targets have been developed to treat this disease. Due to the importance of Cysteine Protease (CP) to several cellular processes in cancer cells, CP inhibitors have been studied as novel alternative approaches for pancreatic cancer therapy. Objective: The cytostatic potential of new CP inhibitors derived from dipeptidyl nitriles is analyzed in vitro using pancreatic cancer (MIA PaCa-2) cells. Methods: The cytotoxic and cytostatic activities were studied using MTT colorimetric assay in 2D and 3D cultures. Colony formation, migration in Boyden chamber and cell cycle analysis were applied to further study the cytostatic activity. The inhibition of cysteine proteases was evaluated with Z-FR-MCA selective substrate, and ROS evaluation was performed with DCFH-DA fluorophore. Permeability was investigated using HPLC-MS to obtain log k(w). Combination therapy was also evaluated using the best compound with gemcitabine. Results: The inhibition of intracellular CP activity by the compounds was confirmed, and the cytostatic effect was established with cell cycle retention in the G1 phase. CP inhibitors were able to reduce cell proliferation by 50% in the clonogenic assay, and the same result was achieved for the migration assay, without any cytotoxic effect. The Neq0554 inhibitor was also efficient to increase the gemcitabine potency in the combination therapy. Physicochemical properties using an artificial membrane model quantified 1.14 >= log K-w >= 0.75 for all inhibitors (also confirmed using HPLC-MS analysis) along with the identification of intra and extracellular metabolites. Finally, these dipeptidyl nitrile derivatives did not trigger the formation of reactive oxygen species, which is linked to genotoxicity. Conclusion: Altogether, these results provide a clear and favorable picture to develop CP inhibitors in pre-clinical assays. (AU)

• Processo FAPESP: 13/18009-4 - Planejamento, síntese e atividade tripanossomicida de inibidores covalentes reversíveis da enzima cruzaína
• Beneficiário: Carlos Alberto Montanari
• Linha de fomento: Auxílio à Pesquisa - Temático
• Processo FAPESP: 14/07292-0 - Planejamento molecular, atividade tripanossomicida e anticancerígena de inibidores covalentes reversíveis de cisteíno proteases
• Beneficiário: José Carlos Quilles Junior
• Linha de fomento: Bolsas no Brasil - Doutorado