Metal Ion-gemcitabine Monophosphate Nanoparticles for Effective Treatment of Pancreatic Cancer

doi:10.6023/A24030085

Abstract

Gemcitabine is the first-line chemotherapeutic agent and the gold standard for pancreatic cancer. However, the short blood half-life and drug resistance limit its therapeutic efficacy in clinic. Recent advancements in nanotechnology offered a potent approach for improving gemcitabine delivery and efficacy. In this study, we present the design and synthesis of gemcitabine-loaded nanoparticles utilizing gemcitabine monophosphate (GMP) and metal ions via a coordination-precipitation strategy. Firstly, a series of metal ions, including Ca²⁺, Fe³⁺, Mn²⁺, Gd³⁺ and Lu³⁺, were mixed with GMP to form nanoparticles (Metal-GMP NPs) employing a reverse-phase microemulsion method. The morphology of the as-prepared Metal-GMP NPs were characterized by transmission electron microscopy while the hydrated particle size, polydispersion index and zeta potential were measured by dynamic light scattering. Additionally, the encapsulation efficiency and loading ratio of GMP, as well as the molar ratio of GMP to metals were quantified through high performance liquid chromatography (HPLC) and inductively coupled plasma mass spectrometry (ICP-MS). Subsequently, the anti-tumor efficacy of the as-prepared Metal-GMP NPs were studied by CCK8 assay on three different pancreatic cancer cell lines. The screening results suggested that Fe-GMP NPs has the preferable anti-tumor activity as well as the obvious synergistic effect between metal ions and drugs. The further mechanism studies revealed that Fe-GMP NPs could generate enough cytotoxic reactive oxygen species (ROS) through Fenton-like reactions within tumor cells. And simultaneously, the intracellular reduced glutathione (GSH) was consumed, thereby disrupting the redox homeostasis and enhancing GEM efficacy. Finally, the therapeutic efficacy and bio-safety of Fe-GMP NPs were investigated on the subcutaneous tumor-bearing model. The in vivo results demonstrated that Fe-GMP NPs exhibited better therapeutic efficacy rather than GMP free drugs alone. Meanwhile, the bio-safety assessment confirmed the absence of significant toxicity or side effects associated with Fe-GMP NP treatment. This work combines metal ions with gemcitabine in the manner of chemistry, providing a highly promising strategy for the delivery and sensitization of gemcitabine in pancreatic cancer therapy.

Key words: gemcitabine monophosphate, metal ion, nanoparticle, pancreatic cancer, synergistic chemotherapy

Cite this article

Qianyu Luo, Chengyan Wang, Tianlong Zhang, Peiyuan Xia, Xiao Zhang, Ming Yang. Metal Ion-gemcitabine Monophosphate Nanoparticles for Effective Treatment of Pancreatic Cancer[J]. Acta Chimica Sinica, 2024, 82(7): 772-781.

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Fig. & Tab. 7

Figure 1. The schematic illustration of preparing Metal-GMP NPs

Figure 2. The Characterization of Metal-GMP NPs. (a) TEM images of different Metal-GMP NPs. (b) The digit photo of Metal-GMP NPs dispersed in water. (c) Hydrodynamic size distributions of Metal-GMP NPs in water. (d) Zeta potentials of Metal-GMP NPs dispersions. (e) GMP Encapsulation efficiency of Metal-GMP NPs. (f) Drug loading capacity of GMP in different Metal-GMP NPs. (g) Molar ratio of metal to GMP in Metal-GMP NPs

Figure 3. Cytotoxicity of Metal-GMP NPs. Cell viability of (a) Aspc-1, (b) Panc-1 and (c) KPC-GR cells after incubating with different Metal-GMP NPs. (d) The combination index (CI) of Metal-GMP NPs

Figure 4. Mechanism investigation for the chemodynamic process of Fe-GMP NPs. (a) Schematic illustration of the Fenton-like catalytic reaction of Fe-GMP NPs. (b) GSH consumption after treated with different concentration of Fe-GMP NPs. (c) Generation of Fe2+ detected by 1,10-phenanthroline. (d) UV-Vis absorption spectra of MB after different treatment. (e) UV-Vis absorption spectra of TMB after different treatment

Figure 5. The cytotoxicity study of Fe-GMP NPs in vitro. (a) Confocal microscope images of Panc-1 cells after treatment with Cy5.5 labeled Fe-GMP NPs. The scale bar is 10 μm. (b) Cellular GSH in Panc-1 cells after different treatment. *P<0.05, **P<0.01. (c) Generation of ROS detected by DCFH-DA. The scale bar is 100 μm. (d) Western blotting of GPX4 protein in Panc-1 cells. (e) Apoptosis assay of Panc-1 cells flow cytometry

Figure 6. In vivo antitumor efficacy of Fe-GMP NPs. (a) Schematic illustration of the establishment of animal model and the anti-tumor experiment. (b) Bioluminescence images of tumor-bearing mice after treatment. (c) Tumor volume profiles in different groups. (d) Tumor weights of mice after treatment. (e) H&E and Tunel staining images of tumor tissue slices. The scale bars are 100 μm. *P<0.05

Figure 7. The bio-safety evaluation of Fe-GMP NPs. (a) Digital photo and quantitative analysis of hemolysis tests. (b) Blood cell analysis and blood biochemical test of the sample from mice after receiving i.v. injection of free GMP and Fe-GMP NPs. (c) H&E staining of the major organs from mice after different treatment. The scale bar is 100 μm

References 29

[1]	Siegel R. L.; Miller K. D.; Wagle N. S.; Jemal A. Ca-Cancer J. Clin. 2023, 73, 17.
[2]	Strobel O.; Neoptolemos J.; Jäger D.; Büchler M. W. Nat. Rev. Clin. Oncol. 2018, 16, 11.
[3]	Saung M. T.; Zheng L. Clin. Ther. 2017, 39, 2125. doi: S0149-2918(17)30904-9 pmid: 28939405
[4]	Gu Z. Y.; Du Y. X.; Zhao X.; Wang C. Cancer Lett. 2021, 521, 98.
[5]	Dubey R. D.; Saneja A.; Gupta P. K.; Gupta P. N. Eur. J. Pharm. Sci. 2016, 93, 147.
[6]	Saiki Y.; Yoshino Y.; Fujimura H.; Manabe T.; Kudo Y.; Shimada M.; Mano N.; Nakano T.; Lee Y.; Shimizu S.; Oba S.; Fujiwara S.; Shimizu H.; Chen N.; Nezhad Z. K.; Jin G.; Fukushige S.; Sunamura M.; Ishida M.; Motoi F.; Egawa S.; Unno M.; Horii A. Biochem. Biophys. Res. Commun. 2012, 421, 98.
[7]	Yamamoto M.; Sanomachi T.; Suzuki S.; Uchida H.; Yonezawa H.; Higa N.; Takajo T.; Yamada Y.; Sugai A.; Togashi K.; Seino S.; Okada M.; Sonoda Y.; Hirano H.; Yoshimoto K.; Kitanaka C. Neuro-oncol. 2021, 23, 945.
[8]	Li X.; Porcel E.; Menendez‐Miranda M.; Qiu J. W.; Yang X. M.; Serre C.; Pastor A.; Desmaële D.; Lacombe S.; Gref R. ChemMedChem 2019, 15, 274.
[9]	Das M.; Li J.; Bao M.; Huang L. AAPS J. 2020, 22, 88.
[10]	Sun H.; Cai H.; Xu C.; Zhai H. Z.; Lux F.; Xie Y.; Feng L.; Du L. Q.; Liu Y.; Sun X. H.; Wang Q.; Song H. J.; He N. N.; Zhang M. M.; Ji K. H.; Wang J. J.; Gu Y. Q.; Leduc G.; Doussineau T.; Wang Y.; Liu Q.; Tillement O. J. Nanobiotechnol. 2022, 20, 449.
[11]	Chen Y.; Huang Y. K.; Zhou S. L.; Sun M. L.; Chen L.; Wang J. H.; Xu M. J.; Liu S. S.; Liang K. F.; Zhang Q.; Jiang T. Z.; Song Q. X.; Jiang G.; Tang X. J.; Gao X. L.; Chen J. Nano Lett. 2020, 20, 6780. doi: 10.1021/acs.nanolett.0c02622 pmid: 32809834
[12]	Lv Y. G.; Chen X.; Shen Y. P. Carbohydr. Polym. 2024, 323, 121434.
[13]	Cai Z.; Zhang Y. W.; Jiang L. P.; Zhu J. J. Acta Chim. Sinica 2021, 79, 481 (in Chinese).
	(蔡政, 张颖雯, 姜立萍, 朱俊杰, 化学学报, 2021, 79, 481.) doi: 10.6023/A20120583
[14]	Li J. J.; Wei J. J.; Gao Y. X.; Zhao Q.; Sun J. J.; Ouyang J.; Na N. Chin. Chem. Lett. 2023, 34, 107662.
[15]	Cordani M.; Resines-Urien E.; Gamonal A.; Milán-Rois P.; Salmon L.; Bousseksou A.; Costa J. S.; Somoza Á. Antioxidants 2021, 10, 66.
[16]	Guo W.; Hu C. Y.; Zhen S. J.; Huang C. Z.; Li Y. F. Acta Chim. Sinica 2022, 80, 1583 (in Chinese). doi: 10.6023/A22070304
	(郭湾, 胡聪意, 甄淑君, 黄承志, 李原芳, 化学学报, 2022, 80, 1583.) doi: 10.6023/A22070304
[17]	Yu X. Y.; Shang T. Y.; Zheng G. D.; Yang H. L.; Li Y. W.; Cai Y. J.; Xie G. X.; Yang B. Chin. Chem. Lett. 2022, 33, 1895.
[18]	Simón M.; Jørgensen J. T.; Khare H. A.; Christensen C.; Nielsen C. H.; Kjaer A. Pharmaceutics 2022, 14, 1284.
[19]	Wang J.; Zhuo L. G.; Zhao P.; Liao W.; Wei H. Y.; Yang Y. C.; Peng S. M.; Yang X. Chin. Chem. Lett. 2022, 33, 3502.
[20]	Man X. Y.; Yang T. F.; Li W. J.; Li S. H.; Xu G.; Zhang Z. L.; Liang H.; Yang F. J. Med. Chem. 2023, 66, 7268.
[21]	Sun H. S.; Zhou J.; Liu C.; Chen X.; Du Y. J.; Li Y. L.; Jiang H.; Wang J. Q.; Song Z.; Guo C. Acta Chim. Sinica 2022, 80, 1250 (in Chinese).
	(孙宏顺, 周进, 刘成, 陈旭, 杜怡璟, 李玉龙, 蒋蕻, 王建强, 宋喆, 郭成, 化学学报, 2022, 80, 1250.) doi: 10.6023/A22050226
[22]	Chou T. C. Cancer Res. 2010, 70, 440.
[23]	Yang Z. B.; Luo Y.; Hu Y. N.; Liang K. C.; He G.; Chen Q.; Wang Q. G.; Chen H. Adv. Funct. Mater. 2020, 31, 2007991.
[24]	Bai Y.; Pan Y. J.; An N.; Zhang H. T.; Wang C.; Tian W.; Huang T. Chin. Chem. Lett. 2023, 34, 107552.
[25]	Dong L. P.; Ding J. S.; Zhu L. M.; Liu Y. J.; Gao X.; Zhou W. H. Chin. Chem. Lett. 2023, 34, 108192.
[26]	Yang Y. H.; Wang Y. F.; Xu L. G.; Chen T. Chin. Chem. Lett. 2020, 31, 1801.
[27]	Raza A.; Hayat U.; Rasheed T.; Bilal M.; Iqbal H. M. N. Eur. J. Med. Chem. 2018, 157, 705. doi: S0223-5234(18)30702-5 pmid: 30138802
[28]	Ouyang J.; Wang L. Q.; Chen W.; Zeng K.; Han Y.; Xu Y.; Xu Q.; Deng L.; Liu Y.-N. Chem. Commun. 2018, 54, 3468.
[29]	Lei G.; Zhuang L.; Gan B. Nat. Rev. Cancer. 2022, 22, 381.

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