Novel nitric oxide donor, nitrated phenylbutyrate, induces cell death of human pancreatic cancer cells and suppresses tumor growth of cancer xenografts

Beppu,Takuro; Beppu,Takuro; Nishi,Koji; Nishi,Koji; Imoto,Shuhei; Imoto,Shuhei; Araki,Waka; Araki,Waka; Setoguchi,Itaru; Setoguchi,Itaru; Ueda,Ayaka; Ueda,Ayaka; Suetsugi,Naho; Suetsugi,Naho; Ishima,Yu; Ishima,Yu; Ikeda,Tokunori; Ikeda,Tokunori; Otagiri,Masaki; Otagiri,Masaki; Yamasaki,Keishi; Yamasaki,Keishi

doi:10.3892/or.2022.8393

Novel nitric oxide donor, nitrated phenylbutyrate, induces cell death of human pancreatic cancer cells and suppresses tumor growth of cancer xenografts

Authors:
- Takuro Beppu
- Koji Nishi
- Shuhei Imoto
- Waka Araki
- Itaru Setoguchi
- Ayaka Ueda
- Naho Suetsugi
- Yu Ishima
- Tokunori Ikeda
- Masaki Otagiri
- Keishi Yamasaki
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Affiliations: Faculty of Pharmaceutical Sciences, Sojo University, Nishi‑ku, Kumamoto 860‑0082, Japan, Department of Pharmacokinetics and Biopharmaceutics, Institute of Biomedical Sciences, Tokushima University, Tokushima 770‑8505, Japan
Published online on: August 23, 2022 https://doi.org/10.3892/or.2022.8393
Article Number: 178

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Abstract

Pancreatic cancer has a low response rate to chemotherapy due to the low drug transferability caused by the low blood flow around the tumor. In the present study, focusing on nitric oxide (NO) for its vasodilatory and antitumor effects, a novel NO donor, a nitrated form of phenylbutyrate (NPB) was synthesized and the antitumor effect on human pancreatic cancer cells (AsPC1 and BxPC3) and xenografts was examined. Using Annexin V, NPB was confirmed to induce cell death against AsPC1 and BxPC3 in a time‑ and concentration‑dependent manner. In NPB‑exposed cells, DAF‑FM DA (a probe to detect intracellular NO) derived fluorescence was observed. Release of nitrite and nitrate from NPB in aqueous solution was very gradual until even 72 h after dissolution. Phenylbutyrate (PB) and hydroxy PB in which the nitro group of NPB was replaced with a hydroxyl group did not have the cell death‑inducing effect as observed in NPB. These results suggest that the effect of NPB was dependent on NO release form NPB. Apoptosis inhibitor, Z‑VAD FMK, had no effect on the cell death‑inducing effect of NPB, and NPB did not show significant activation of caspase‑3/7. In addition, NPB significantly decreased cellular ATP levels, suggesting that necrosis is involved in the effect of NPB. NPB also accumulated cells specifically at the S phase of the cell cycle. A single dose of NPB (10 mg/kg) into mice with established BxPC3 xenografts significantly suppressed tumor growth for at least 7 weeks without apparent toxicity. The findings of the present study indicate that NPB has potential as a novel therapeutic agent for NO‑based therapy of pancreatic cancer.

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1	Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D and Bray F: Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 136:E359–E386. 2015. View Article : Google Scholar : PubMed/NCBI
2	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2015. CA Cancer J Clin. 65:5–29. 2015. View Article : Google Scholar : PubMed/NCBI
3	Le Large TY, Bijlsma MF, Kazemier G, van Laarhoven HW, Giovannetti E and Jimenez CR: Key biological processes driving metastatic spread of pancreatic cancer as identified by multi-omics studies. Semin Cancer Biol. 44:153–169. 2017. View Article : Google Scholar : PubMed/NCBI
4	Ercan G, Karlitepe A and Ozpolat B: Pancreatic cancer stem cells and therapeutic approaches. Anticancer Res. 37:2761–2775. 2017.PubMed/NCBI
5	Hwang RF, Moore T, Arumugam T, Ramachandran V, Amos KD, Rivera A, Ji B, Evans DB and Logsdon CD: Cancer-associated stromal fibroblasts promote pancreatic tumor progression. Cancer Res. 68:918–926. 2008. View Article : Google Scholar : PubMed/NCBI
6	Rees DD, Palmer RM and Moncada S: Role of endothelium-derived nitric oxide in the regulation of blood pressure. Proc Natl Acad Sci USA. 86:3375–3378. 1989. View Article : Google Scholar : PubMed/NCBI
7	Loscalzo J: Nitric oxide insufficiency, platelet activation, and arterial thrombosis. Circ Res. 88:756–762. 2001. View Article : Google Scholar : PubMed/NCBI
8	Korhonen R, Lahti A, Kankaanranta H and Moilanen E: Nitric oxide production and signaling in inflammation. Curr Drug Targets Inflamm Allergy. 4:471–479. 2005. View Article : Google Scholar : PubMed/NCBI
9	Williams JL, Borgo S, Hasan I, Castillo E, Traganos F and Rigas B: Nitric oxide-releasing nonsteroidal anti-inflammatory drugs (NSAIDs) alter the kinetics of human colon cancer cell lines more effectively than traditional NSAIDs: Implications for colon cancer chemoprevention. Cancer Res. 61:3285–3289. 2001.PubMed/NCBI
10	Pervin S, Singh R, Gau CL, Edamatsu H and Tamanoi F: Potentiation of nitric oxide-induced apoptosis of MDA-MB-468 cells by farnesyltransferase inhibitor: Implications in breast cancer. Cancer Res. 61:4701–4706. 2001.PubMed/NCBI
11	Yang L, Lan C, Fang Y, Zhang Y, Wang J, Guo J, Wan S, Yang S, Wang R and Fang D: Sodium nitroprusside (SNP) sensitizes human gastric cancer cells to TRAIL-induced apoptosis. Int Immunopharmacol. 17:383–389. 2013. View Article : Google Scholar : PubMed/NCBI
12	Mitrovic B, Ignarro LJ, Vinters HV, Akers MA, Schmid I, Uittenbogaart C and Merrill JE: Nitric oxide induces necrotic but not apoptotic cell death in oligodendrocytes. Neuroscience. 65:531–539. 1995. View Article : Google Scholar : PubMed/NCBI
13	Kang F, Zhu J, Wu J, Lv T, Xiang H, Tian J, Zhang Y and Huang Z: O 2-3-Aminopropyl diazeniumdiolates suppress the progression of highly metastatic triple-negative breast cancer by inhibition of microvesicle formation via nitric oxide-based epigenetic regulation. Chem Sci. 9:6893–6898. 2018. View Article : Google Scholar : PubMed/NCBI
14	Kinoshita R, Ishima Y, Ikeda M, Kragh-Hansen U, Fang J, Nakamura H, Chuang VT, Tanaka R, Maeda H, Kodama A, et al: S-Nitrosated human serum albumin dimer as novel nano-EPR enhancer applied to macromolecular anti-tumor drugs such as micelles and liposomes. J Control Release. 217:1–9. 2015. View Article : Google Scholar : PubMed/NCBI
15	Enokida T, Yamasaki K, Okamoto Y, Taguchi K, Ishiguro T, Maruyama T, Seo H and Otagiri M: Tyrosine411 and Arginine410 of human serum albumin play an important role in the binding of sodium 4-phenylbutyrate to site II. J Pharm Sci. 105:1987–1994. 2016. View Article : Google Scholar : PubMed/NCBI
16	Krach-Hansen U, Chuang VT and Otagiri M: Practical aspects of the ligand-binding and enzymatic properties of human serum albumin. Biol Pharm Bull. 25:695–704. 2002. View Article : Google Scholar : PubMed/NCBI
17	Heise T and Mathieu C: Impact of the mode of protraction of basal insulin therapies on their pharmacokinetic and pharmacodynamic properties and resulting clinical outcomes. Diabetes Obes Metab. 19:3–12. 2017. View Article : Google Scholar : PubMed/NCBI
18	Heinemann L, Sinha K, Weyer C, Loftager M, Hirschberger S and Heise T: Time-action profile of the soluble, fatty acid acylated, long-acting insulin analogue NN304. Diabet Med. 16:332–338. 1999. View Article : Google Scholar : PubMed/NCBI
19	Knudsen LB, Nielsen PF, Huusfeldt PO, Johansen NL, Madsen K, Pedersen FZ, Thøgersen H, Wilken M and Agersø H: Potent derivatives of glucagon-like peptide-1 with pharmacokinetic properties suitable for once daily administration. J Med Chem. 43:1664–1669. 2000. View Article : Google Scholar : PubMed/NCBI
20	Matsumura Y and Maeda H: A new concept for macromolecular therapeutics in cancer chemotherapy: Mechanism of tumoritropic accumulation of proteins and the antitumor agent smancs. Cancer Res. 46:6387–6392. 1986.PubMed/NCBI
21	Durante M, Frosini M, Fusi F, Neri A, Sticozzi C and Saponara S: In vitro vascular toxicity assessment of NitDOX, a novel NO-releasing doxorubicin. Eur J Pharmacol. 880:1731642020. View Article : Google Scholar : PubMed/NCBI
22	Maeda H, Akaike T, Yoshida M and Suga M: Multiple functions of nitric oxide in pathophysiology and microbiology: Analysis by a new nitric oxide scavenger. J Leukoc Biol. 56:588–592. 1994. View Article : Google Scholar : PubMed/NCBI
23	Huang Z, Liu L, Chen J, Cao M and Wang J: JS-K as a nitric oxide donor induces apoptosis via the ROS/Ca2+/caspase-mediated mitochondrial pathway in HepG2 cells. Biomed Pharmacother. 107:1385–1392. 2018. View Article : Google Scholar : PubMed/NCBI
24	Millet A, Bettaieb A, Renaud F, Prevotat L, Hammann A, Solary E, Mignotte B and Jeannin JF: Influence of the nitric oxide donor glyceryl trinitrate on apoptotic pathways in human colon cancer cells. Gastroenterology. 123:235–246. 2002. View Article : Google Scholar : PubMed/NCBI
25	Liu L, Li T, Tan J, Fu J, Guo Q, Ji H and Zhang Y: NG as a novel nitric oxide donor induces apoptosis by increasing reactive oxygen species and inhibiting mitochondrial function in MGC803 cells. Int Immunopharmacol. 23:27–36. 2014. View Article : Google Scholar : PubMed/NCBI
26	Kroemer G, Galluzzi L, Vandenabeele P, Abrams J, Alnemri ES, Baehrecke EH, Blagosklonny MV, El-Deiry WS, Golstein P, Green DR, et al: Classification of cell death: Recommendations of the nomenclature committee on cell death 2009. Cell Death Differ. 16:3–11. 2009. View Article : Google Scholar : PubMed/NCBI
27	Eiserich JP, Patel RP and O'Donnell VB: Pathophysiology of nitric oxide and related species: Free radical reactions and modification of biomolecules. Mol Aspects Med. 19:221–357. 1998. View Article : Google Scholar : PubMed/NCBI
28	Gotoh T and Mori M: Nitric oxide and endoplasmic reticulum stress. Arterioscler Thromb Vasc Biol. 26:1439–1446. 2006. View Article : Google Scholar : PubMed/NCBI
29	Kashfi K, Rayyan Y, Qiao LL, Williams JL, Chen J, Del Soldato P, Traganos F, Rigas B and Ryann Y: Nitric oxide-donating nonsteroidal anti-inflammatory drugs inhibit the growth of various cultured human cancer cells: Evidence of a tissue type-independent effect. J Pharmacol Exp Ther. 303:1273–1282. 2002. View Article : Google Scholar : PubMed/NCBI
30	Dunlap T, Abdul-Hay SO, Chandrasena RE, Hagos GK, Sinha V, Wang Z, Wang H and Thatcher GR: Nitrates and NO-NSAIDs in cancer chemoprevention and therapy: In vitro evidence querying the NO donor functionality. Nitric Oxide. 19:115–124. 2008. View Article : Google Scholar : PubMed/NCBI
31	Wu W, Gaucher C, Fries I, Hu XM, Maincent P and Sapin-Minet A: Polymer nanocomposite particles of S-nitrosoglutathione: A suitable formulation for protection and sustained oral delivery. Int J Pharm. 495:354–361. 2015. View Article : Google Scholar : PubMed/NCBI
32	Yang C, Hwang HH, Jeong S, Seo D, Jeong Y, Lee DY and Lee K: Inducing angiogenesis with the controlled release of nitric oxide from biodegradable and biocompatible copolymeric nanoparticles. Int J Nanomedicine. 13:6517–6530. 2018. View Article : Google Scholar : PubMed/NCBI
33	Laschak M, Spindler KD, Schrader AJ, Hessenauer A, Streicher W, Schrader M and Cronauer MV: JS-K, a glutathione/glutathione S-transferase-activated nitric oxide releasing prodrug inhibits androgen receptor and WNT-signaling in prostate cancer cells. BMC Cancer. 12:1302012. View Article : Google Scholar : PubMed/NCBI
34	Nishi K, Imoto S, Beppu T, Uchibori S, Yano A, Ishima YU, Ikeda T, Tsukigawa K, Otagiri M and Yamasaki K: The nitrated form of nateglinide induces apoptosis in human pancreatic cancer cells through a caspase-dependent mechanism. Anticancer Res. 42:1333–1338. 2022. View Article : Google Scholar : PubMed/NCBI