In vitro evaluation of novel N-acetylalaninate prodrugs that selectively induce apoptosis in prostate cancer cells

BMC Cancer

Christopher A McGoldrick¹  Yu-Lin Jiang²  Marianne Brannon¹  Koyamangalath Krishnan³  William L Stone¹ 

[1] Department of Pediatrics, East Tennessee State University, Johnson City, TN 37614-0578, USA

[2] Center for Biomedical Imaging, Department of Radiology and Radiological Sciences, Medical University of South Carolina, Charleston, SC 29425, USA

[3] Division of Hematology-Oncology, Department of Internal Medicine, East Tennessee State University, Johnson City, TN 37614, USA

关键词: Quinone methide;    Reactive oxygen species;    Oxidized protein hydrolase;    Cell viability;    Apoptosis;    Oxidative stress;    Glutathione;    Chemotherapy;    Prodrugs;    Prostate cancer;   

Others  :  1121066
DOI  :  10.1186/1471-2407-14-675

 received in 2014-05-07, accepted in 2014-09-16,  发布年份 2014

【 摘 要 】

Background

Cancer cell esterases are often overexpressed and can have chiral specificities different from that of the corresponding normal cells and can, therefore, be useful targets for activating chemotherapeutic prodrug esters. Prodrug esters are inactive compounds that can be preferentially activated by esterase enzymes. Moreover, cancer cells often exhibit a high level of intrinsic oxidative stress due to an increased formation of reactive oxygen species (ROS) and a decreased expression of some enzymatic antioxidants. Prodrugs designed to induce additional oxidative stress can selectively induce apoptosis in cancer cells already exhibiting a high level of intrinsic oxidative stress. This study focused on the in vitro evaluation of four novel prodrug esters: the R- and S- chiral esters of 4-[(nitrooxy)methyl]phenyl N-acetylalaninate (R- and S-NPAA) and the R- and S- chiral esters of 4-[(nitrooxy)methyl]naphth-1-yl N-acetylalaninate (R- and S-NQM), which are activated, to varying extents, by oxidized protein hydrolase (OPH, EC 3.4.19.1) yielding a quinone methide (QM) intermediate capable of depleting glutathione (GSH), a key intracellular antioxidant. OPH is a serine esterase/protease that is overexpressed in some human tumors and cancer cell lines.

Methods

To evaluate the chiral ester prodrugs, we monitored cellular GSH depletion, cellular protein carbonyl levels (an oxidative stress biomarker) and cell viability in tumorigenic and nontumorigenic prostate cancer cell lines.

Results

We found that the prodrugs were activated by OPH and subsequently depleted GSH. The S-chiral ester of NPAA (S-NPAA) was two-fold more effective than the R-chiral ester (R-NPAA) in depleting GSH, increasing oxidative stress, inducing apoptosis, and decreasing cell viability in tumorigenic prostate LNCaP cells but had little effect on non-tumorigenic RWPE-1 cells. In addition, we found that that S-NPAA induced apoptosis and decreased cell viability in tumorigenic DU145 and PC3 prostate cell lines. Similar results were found in a COS-7 model that overexpressed active human OPH (COS-7-OPH).

Conclusions

Our results suggest that prostate tumors overexpressing OPH and/or exhibiting a high level of intrinsic oxidative stress may be susceptible to QM generating prodrug esters that are targeted to OPH with little effect on non-tumorigenic prostate cells.

【 授权许可】

   
2014 McGoldrick et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Nogueira V, Park Y, Chen CC, Xu PZ, Chen ML, Tonic I, Unterman T, Hay N: Akt determines replicative senescence and oxidative or oncogenic premature senescence and sensitizes cells to oxidative apoptosis. Cancer Cell 2008, 14(6):458-470.
• [2]Pelicano H, Carney D, Huang P: ROS stress in cancer cells and therapeutic implications. Drug Resist Updat 2004, 7(2):97-110.
• [3]Kumar B, Koul S, Khandrika L, Meacham RB, Koul HK: Oxidative stress is inherent in prostate cancer cells and is required for aggressive phenotype. Cancer Res 2008, 68(6):1777-1785.
• [4]Hileman EO, Liu J, Albitar M, Keating MJ, Huang P: Intrinsic oxidative stress in cancer cells: a biochemical basis for therapeutic selectivity. Cancer Chemother Pharmacol 2004, 53(3):209-219.
• [5]Dolado I, Nebreda AR: AKT and oxidative stress team up to kill cancer cells. Cancer Cell 2008, 14(6):427-429.
• [6]Govindarajan B, Sligh JE, Vincent BJ, Li M, Canter JA, Nickoloff BJ, Rodenburg RJ, Smeitink JA, Oberley L, Zhang Y, Slingerland J, Arnold RS, Lambeth JD, Cohen C, Hilenski L, Griendling K, Martinez-Diez M, Cuezva JM, Arbiser JL: Overexpression of Akt converts radial growth melanoma to vertical growth melanoma. J Clin Invest 2007, 117(3):719-729.
• [7]Watson J: Oxidants, antioxidants and the current incurability of metastatic cancers. Open Biol 2013, 3(1):120144.
• [8]Testa JR, Tsichlis PN: AKT signaling in normal and malignant cells. Oncogene 2005, 24(50):7391-7393.
• [9]Benbrook DM, Masamha CP: The pro-survival function of Akt kinase can be overridden or altered to contribute to induction of apoptosis. Curr Cancer Drug Targets 2011, 11(5):586-599.
• [10]Singh S, Khan AR, Gupta AK: Role of glutathione in cancer pathophysiology and therapeutic interventions. J Exp Ther Oncol 2012, 9(4):303-316.
• [11]Mena S, Benlloch M, Ortega A, Carretero J, Obrador E, Asensi M, Petschen I, Brown BD, Estrela JM: Bcl-2 and glutathione depletion sensitizes B16 melanoma to combination therapy and eliminates metastatic disease. Clin Cancer Res 2007, 13(9):2658-2666.
• [12]Mena S, Rodriguez ML, Ortega A, Priego S, Obrador E, Asensi M, Petschen I, Cerda M, Brown BD, Estrela JM: Glutathione and Bcl-2 targeting facilitates elimination by chemoradiotherapy of human A375 melanoma xenografts overexpressing bcl-xl, bcl-2, and mcl-1. J Transl Med 2012, 10(1):8. BioMed Central Full Text
• [13]Schnelldorfer T, Gansauge S, Gansauge F, Schlosser S, Beger HG, Nussler AK: Glutathione depletion causes cell growth inhibition and enhanced apoptosis in pancreatic cancer cells. Cancer 2000, 89(7):1440-1447.
• [14]Dunlap T, Chandrasena RE, Wang Z, Sinha V, Wang Z, Thatcher GR: Quinone formation as a chemoprevention strategy for hybrid drugs: balancing cytotoxicity and cytoprotection. Chem Res Toxicol 2007, 20(12):1903-1912.
• [15]Gao J, Liu X, Rigas B: Nitric oxide-donating aspirin induces apoptosis in human colon cancer cells through induction of oxidative stress. Proc Natl Acad Sci U S A 2005, 102(47):17207-17212.
• [16]Gao L, Williams JL: Nitric oxide-donating aspirin induces G2/M phase cell cycle arrest in human cancer cells by regulating phase transition proteins. Int J Oncol 2012, 41(1):325-330.
• [17]Khan NI, Cisterne A, Baraz R, Bradstock KF, Bendall LJ: Para-NO-aspirin inhibits NF-kappaB and induces apoptosis in B-cell progenitor acute lymphoblastic leukemia. Exp Hematol 2012, 40(3):207-215.e1.
• [18]Razavi R, Gehrke I, Gandhirajan RK, Poll-Wolbeck SJ, Hallek M, Kreuzer KA: Nitric oxide-donating acetylsalicylic acid induces apoptosis in chronic lymphocytic leukemia cells and shows strong antitumor efficacy in vivo. Clin Cancer Res 2011, 17(2):286-293.
• [19]Rigas B: The use of nitric oxide-donating nonsteroidal anti-inflammatory drugs in the chemoprevention of colorectal neoplasia. Curr Opin Gastroenterol 2007, 23(1):55-59.
• [20]Sun Y, Chen J, Rigas B: Chemopreventive agents induce oxidative stress in cancer cells leading to COX-2 overexpression and COX-2-independent cell death. Carcinogenesis 2009, 30(1):93-100.
• [21]Sun Y, Rigas B: The thioredoxin system mediates redox-induced cell death in human colon cancer cells: implications for the mechanism of action of anticancer agents. Cancer Res 2008, 68(20):8269-8277.
• [22]Hulsman N, Medema JP, Bos C, Jongejan A, Leurs R, Smit MJ, de Esch IJP, Richel D, Wijtmans M: Chemical insights in the concept of hybrid drugs: the antitumor effect of nitric oxide-donating aspirin involves a Quinone methide but Not nitric oxide nor aspirin. J Med Chem 2007, 50(10):2424-2431.
• [23]Stone WL, Jiang Yu L, McGoldrick C, Brannon M, Krishnan K: The Design, Synthesis and in Vitro Evaluation of a Novel pro-Oxidant Anticancer Prodrug Substrate Targeted to Acylamino-Acid Releasing Enzyme. In Free Radicals: The Role of Antioxidants and pro-Oxidants in Cancer Development. 1st edition. Edited by Stone WL. New York: Nova Biomedical; 2014:189.
• [24]McGoldrick CA, Jiang YL, Paromov V, Brannon M, Krishnan K, Stone WL: Identification of oxidized protein hydrolase as a potential prodrug target in prostate cancer. BMC Cancer 2014., 14(1) 77-2407-14-77
• [25]http://www.proteinatlas.org/ENSG00000164062/cancer
• [26]Mautjana NA, Looi DW, Eyler JR, Brajter-Toth A: Sensitivity of positive Ion mode electrospray ionization mass spectrometry in the analysis of thiol metabolites. Electroanalysis 2010, 22(1):79-89.
• [27]Enari M, Sakahira H, Yokoyama H, Okawa K, Iwamatsu A, Nagata S: A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 1998, 391(6662):43-50.
• [28]Schumacker PT: Reactive oxygen species in cancer cells: live by the sword, die by the sword. Cancer Cell 2006, 10(3):175-176.
• [29]Mitchell JB, Russo A: The role of glutathione in radiation and drug induced cytotoxicity. Br J Cancer Suppl 1987, 8:96-104.
• [30]Diehn M, Cho RW, Lobo NA, Kalisky T, Dorie MJ, Kulp AN, Qian D, Lam JS, Ailles LE, Wong M, Joshua B, Kaplan MJ, Wapnir I, Dirbas FM, Somlo G, Garberoglio C, Paz B, Shen J, Lau SK, Quake SR, Brown JM, Weissman IL, Clarke MF: Association of reactive oxygen species levels and radioresistance in cancer stem cells. Nature 2009, 458(7239):780-783.
• [31]Abdalla MY: Glutathione as potential target for cancer therapy; more or less is good? (mini-review). Jordan J Biol Sci 2011, 4:119.