5-Fluorocytosine–Sugar Conjugates for Glucose Transporter-Mediated Tumor Targeting: Synthesis, Cytotoxicity, and Cellular Uptake Mechanism

Abstract

Two novel sugar-conjugated 5-fluorocytosine (5-FC) antineoplastic compounds were designed and synthesized to improve the selective drug uptake by targeting the tumor-specific glucose transporter (GLUT). The antitumor activity of these compounds was evaluated in four different human cancer cell lines: A549 (human lung cancer cell line), HT29 (human colorectal cancer cell line), H460 (human lung cancer cell line), and PC3 (human prostate cancer cell line). The sugar conjugates exhibited cytotoxicity similar to or higher than 5-FC and 1-hexylcarbamoyl-5-FC in A549, HT29, H460, and PC3. Furthermore, GLUT-mediated transport of the glycoconjugate was investigated with GLUT inhibitor-mediated cytotoxicity analysis in a GLUT-overexpressing HT29 cell line. The cell-killing potency of 5-FC glycoconjugate was found to depend significantly on the GLUT inhibitor, and the cellular uptake of molecules was regulated by GLUT-mediated transport. All the results demonstrate the potential advantages of glycoconjugation for Warburg effect-targeted drug design.

Keywords

Warburg effect / Glucose transporter overexpressed / 5-Fluorocytosine glycoconjugate / Tumor targeting

References

[1] Warburg O. On the origin of cancer cells. Science, 1956, 123(3191): 309-314.

[2] Warburg O, Wind F, Negelein E. The metabolism of tumors in the body. J Gen Physiol, 1927, 8(6): 519-530.

[3] Boado RJ, Black KL, Pardridge WM. Gene expression of GLUT3 and GLUT1 glucose transporters in human brain tumors. Mol Brain Res, 1994, 27(1): 51-57.

[4] Godoy A, Ulloa V, Rodrıguez F, et al. Differential subcellular distribution of glucose transporters GLUT1-6 and GLUT9 in human cancer: ultrastructural localization of GLUT1 and GLUT5 in breast tumor tissues. J Cell Physiol, 2006, 207(3): 614-627.

[5] Kurata T, Oguri T, Isobe T, et al. Differential expression of facilitative glucose transporter (GLUT) genes in primary lung cancers and their liver metastases. Jpn J Cancer Res, 1999, 90(11): 1238-1243.

[6] Younes M, Brown RW, Stephenson M, et al. Overexpression of Glut1 and Glut3 in stage I nonsmall cell lung carcinoma is associated with poor survival. Cancer, 1997, 80(6): 1046-1051.

[7] Plathow C, Weber WA. Tumor cell metabolism imaging. J Nucl Med, 2008, 49(Suppl 2): 43S-63S.

[8] Pohl J, Bertram B, Hilgard P, et al. D-19575: a sugar-linked isophosphoramide mustard derivative exploiting transmembrane glucose transport. Cancer Chemother Pharmacol, 1995, 35(5): 364-370.

[9] Schechter NR, Erwin WD, Yang DJ, et al. Radiation dosimetry and biodistribution of ^99mTc-ethylene dicysteine-deoxyglucose in patients with non-small-cell lung cancer. Eur J Nucl Med Mol Imaging, 2009, 36(10): 1583-1591.

[10] Zhang M, Zhang Z, Blessington D, et al. Pyropheophorbide 2-deoxyglucosamide: a new photosensitizer targeting glucose transporters. Bioconjug Chem, 2003, 14(4): 709-714.

[11] Bronstein Y, Tummala S, Rohren E, et al. F-18 FDG PET/CT for detection of malignant involvement of peripheral nerves. Clin Nucl Med, 2011, 36(2): 96-100.

[12] Ben-Haim S, Ell P. ¹⁸F-FDG PET and PET/CT in the evaluation of cancer treatment response. J Nucl Med, 2009, 50(1): 88-99.

[13] Malet-Martino M, Jolimaitre P, Martino R. The prodrugs of 5-fluorouracil. Curr Med Chem Anti-Cancer Agents, 2002, 2(2): 267-310.

[14] Longley DB, Harkin DP, Johnston PG. 5-Fluorouracil: mechanisms of action and clinical strategies. Nat Rev Cancer, 2003, 3(5): 330-338.

[15] Martino-Echarri E, Henderson BR, Brocardo MG. Targeting the DNA replication checkpoint by pharmacologic inhibition of Chk1 kinase: a strategy to sensitize APC mutant colon cancer cells to 5-fluorouracil chemotherapy. Oncotarget, 2014, 5(20): 9889-9900.

[16] Cassidy J, Saltz L, Twelves C, et al. Efficacy of capecitabine versus 5-fluorouracil in colorectal and gastric cancers: a meta-analysis of individual data from 6171 patients. Ann Oncol, 2011, 22(12): 2604-2609.

[17] Kodama Y, Fumoto S, Nishi J, et al. Absorption and distribution characteristics of 5-fluorouracil (5-FU) after an application to the liver surface in rats in order to reduce systemic side effects. Biol Pharm Bull, 2008, 31(5): 1049-1052.

[18] Suzuki K, Oda H, Sugawara Y, et al. Oxaliplatin-induced acute thrombocytopenia: a case report and review of the literature. Intern Med, 2013, 52(5): 611-615.

[19] Bautista MA, Stevens WT, Chen CS, et al. Hypersensitivity reaction and acute immune-mediated thrombocytopenia from oxaliplatin: two case reports and a review of the literature. J Hematol Oncol, 2010, 3(1): 12-19.

[20] Gao Y, Xiao X, Zhang C, et al. Melatonin synergizes the chemotherapeutic effect of 5-fluorouracil in colon cancer by suppressing PI3 K/AKT and NF-κB/iNOS signaling pathways. J Pineal Res, 2016, 62(2): e12380

[21] Yata VK, Gopinath P, Ghosh SS. Emerging implications of nonmammalian cytosine deaminases on cancer therapeutics. Appl Biochem Biotechnol, 2012, 167(7): 2103-2116.

[22] Liu M, Zhang T, Li D, et al. A first step in the development of gene therapy for colorectal carcinoma: cloning, sequencing, and expression of Escherichia coli cytosine deaminase. Mol Pharmacol, 1993, 43(3): 380-387.

[23] Huber BE, Austin EA, Good S, et al. In vivo antitumor activity of 5-fluorocytosine on human colorectal carcinoma cells genetically modified to express cytosine deaminase. Can Res, 1993, 53(19): 4619-4626.

[24] Corban-Wilhelm H, Ehemann V, Becker G, et al. Comparison of different methods to assess the cytotoxic effects of cytosine deaminase and thymidine kinase gene therapy. Cancer Gene Ther, 2004, 11(3): 208-214.

[25] Gopinath P, Ghosh SS. Understanding apoptotic signaling pathways in cytosine deaminase-uracil phosphoribosyl transferase-mediated suicide gene therapy in vitro. Mol Cell Biochem, 2009, 324(1–2): 21-29.

[26] Lv Z, Zhang TY, Yin JC, et al. Enhancement of anti-tumor activity of newcastle disease virus by the synergistic effect of cytosine deaminase. Asian Pac J Cancer Prev, 2013, 14(12): 7489-7496.

[27] Wang W, Zhang N, Zhao T, et al. Inhibition of tumor growth by polyarginine-fused mutant cytosine deaminase. Appl Biochem Biotechnol, 2015, 175(3): 1633-1643.

[28] Mitchell LA, Espinoza FL, Mendoza D, et al. Toca 511 gene transfer and treatment with the prodrug, 5-fluorocytosine, promotes durable antitumor immunity in a mouse glioma model. Neuro Oncology, 2017, 19(7): 930-939.

[29] Liu PX, Lu YH, Gao XQ, et al. Highly water-soluble platinum(II) complexes as GLUT substrates for targeted therapy: improved anticancer efficacy and transporter-mediated cytotoxic properties. Chem Commun, 2013, 49(24): 2421-2423.

[30] Li H, Gao XQ, Liu R, et al. Glucose conjugated platinum(II) complex: Antitumor superiority to oxaliplatin, combination effect and mechanism of action. Eur J Med Chem, 2015, 101: 400-408.

[31] Wu M, Li H, Liu R, et al. Galactose conjugated platinum(II) complex targeting the Warburg effect for treatment of non-small cell lung cancer and colon cancer. Eur J Med Chem, 2016, 110: 32-42.

[32] Mi Q, Ma Y, Gao X, et al. 2-Deoxyglucose conjugated platinum(II) complexes for targeted therapy: design, synthesis, and antitumor activity. J Biomol Struct Dyn, 2016, 34(11): 2339-2350.

[33] Gao X, Liu S, Shi Y, et al. Mechanistic and biological characteristics of different sugar conjugated 2-methyl malonatoplatinum(II) complexes as new tumor targeting agents. Eur J Med Chem, 2017, 125: 372-384.

[34] Liu R, Li H, Gao X, et al. Mannose-conjugated platinum complexes reveals effective tumor targeting mediated by glucose transporter 1. Biochem Biophy Res Commun, 2017, 487(1): 34-40.

[35] Han J, Gao X, Liu R, et al. Design, synthesis of novel platinum(II) glycoconjugates, and evaluation of their antitumor effects. Chem Biol Drug Des, 2016, 87(6): 867-877.

[36] Ozaki S, Watanabe Y, Nagase T, et al. 5-Fluorouracil derivatives. XI. Synthesis of 1-hexylcarbamoyl-5-fluorouracil metabolites. Chem Pharm Bull, 1986, 34(2): 893-896.

[37] Pizzirani D, Pagliuca C, Realini N, et al. discovery of a new class of highly potent inhibitors of acid ceramidase: synthesis and structure–activity relationship (SAR). J Med Chem, 2013, 56(9): 3518-3530.

[38] Li T, Gao X, Yang L, et al. Methyl 6-amino-6-deoxy-d-pyranoside-conjugated platinum(II) complexes for glucose transporter (GLUT)-mediated tumor targeting: synthesis, cytotoxicity, and cellular uptake mechanism. ChemMedChem, 2016, 11(10): 1069-1077.