Synthesis and Antitumor Activity of Novel Amino Glucose Derivative Schiff Base Copper(II) Complexes

doi:10.6023/A25090314

Abstract

Introducing biologically active organic moieties into the coordination field of transition metal complexes has become an effective strategy for improving the selectivity and biocompatibility of metal drugs. In this paper, two ligands L¹and L² with azidoacetylated glucosamine Schiff base structure were designed and synthesized, and then reacted with copper metal salts to synthesize three novel copper complexes. The single crystal structures of ligand L¹and complex 3 were analyzed by X-ray single crystal diffraction technology, and the in vitro cytotoxicity of the compounds on human malignant tumor cell lines and normal cell lines was determined by methyl thiazolyl tetrazolium (MTT) assay. Among them, complex 2 showed high cytotoxicity against HepG-2 and HeLa cells, approaching or exceeding the value of the positive control, but the toxicity to normal cells was much lower than that of the positive control. The subsequent anti-tumor activity studies of complex 2 with strong cytotoxicity to tumor cells, including apoptosis assessment, intracellular reactive oxygen species generation, mitochondrial membrane potential, cell cycle, cell uptake, migration and invasion, and angiogenesis experiments, further confirmed its inhibitory mechanism on tumor cells. Due to the excellent water solubility of the sugar skeleton in the ligand, complex 2 exhibited good solubility and stability. We further demonstrated through intracellular reactive oxygen species (ROS) production, Hoechst 33342 staining assay, mitochondrial membrane potential assay and Annexin V-mCherry assay that complex 2 can increase the level of ROS in cancer cells and induce apoptosis through the mitochondrial pathway. Cell cycle experiments have shown that complex 2 can delay or inhibit the progression of the cell cycle through the S and G2/M phases. The significant cell uptake results indicated that the high cytotoxicity of complex 2 might be due to its extensive accumulation in HepG-2 cells. In addition, complex 2 can effectively inhibit the migration and invasion of cancer cells, as well as the angiogenesis of normal cell line umbilical vein endothelial cells (HUVEC). Molecular docking results showed that complex 2 has good binding ability with vascular endothelial growth factor receptor 2 (VEGFR-2).

Key words: crystal structures, glucosamine, apoptosis, cellular uptake, migration and invasion

Cite this article

Gao Chunyan, Cui Xinyue, Wang Kaiyue, Shi Xueyi, He Qing, Zhao Jinzhong, Yue Aiqin, Du Weijun, Zhang Yongpo. Synthesis and Antitumor Activity of Novel Amino Glucose Derivative Schiff Base Copper(II) Complexes[J]. Acta Chimica Sinica, 2026, 84(3): 353-363.

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Fig. & Tab. 13

Scheme 1. Synthesis route of ligands L1, L2 and Cu(II) complexes 1~3

Figure 1. Crystal structure diagrams of L1 and complex 3, all hydrogen atoms and external methanol and water solvents were omitted for clarity

Compound	IC₅₀^a/(μmol•L⁻¹)
Compound	HeLa	NCI-H460	BGC-823	HepG-2	HUVEC
Oxaliplatin	48.48±0.22	35.10±0.74	9.16±0.19	12.21±3.17	10.64±3.68
1	29.96±3.11	28.88±4.49	26.34±4.54	28.72±1.98	23.15±2.09
2	22.36±3.96	31.26±0.78	19.20±2.47	13.59±1.00	20.08±1.45
3	48.88±1.12	44.43±2.41	44.83±2.12	30.54±1.24	72.436
L¹	>100	>100	>100	>100	—
L²	>100	>100	>100	>100	—
CuBr₂	161.5	106.0	84.7	117.7	—
CuCl₂	142.0	106.8	88.9	102.3	—

Table 1. IC50 values of the compounds against the tested cell lines for 48 h

Figure 2. The effect of time on the cytotoxicity of the complex 2 against HeLa (a) and HepG-2 (b) cells for 24, 48 and 72 h, respectively

Figure 3. Hoechst 33342 staining to detect the effect of complex 2 (20 μmol/L) on the morphology of HeLa and HepG-2 cells for 24 h and the optical density value of the relative fluorescence intensity (10×10). Note: Data are expressed as mean ± standard deviation of three independent experiments. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns=non-significant

Figure 4. DCFH-DA probe staining to determine the effect of complex 2 on ROS production in HeLa, HepG-2 cells for 24 h, and the optical density value of the relative fluorescence intensity (10×10)

Figure 5. The effect of complex 2 on mitochondrial membrane potentials of HeLa, HepG-2 cells for 24 h were determined with JC-1 and the optical density value of the relative fluorescence intensity (10×10) (a: HeLa cells, b: HepG-2 cells)

Figure 6. Annexin V-mCherry staining detects complex 2 induced apoptosis in HepG-2 cells

Figure 7. Flow cytometry detection of the effect of complex 2 on HepG-2 cell cycle for 24 h

Figure 8. Histogram of copper uptake in HepG-2 cells treated with complex 2 for 24 h, **p<0.01 (left); Statistical data of copper uptake (right)

Figure 9. The effect of complex 2 on HeLa and HepG-2 cells migration and invasion and the number of cells passing through the chamber (10×10). Note: The data is represented as mean ± standard deviation of three independent experiments. **p<0.01, ##p<0.01 (a: HeLa cells, 20 μmol/L Oxaliplatin, 20 μmol/L 2; b: HepG-2 cells, 15 μmol/L Oxaliplatin, 15 μmol/L 2)

Figure 10. The effect of complex 2 (20 μmol/L) on the angiogenesis of HUVEC cells and the number of vascular branches and the total length of vessels (10×4). Note: Data are expressed as mean ± standard deviation of three independent experiments. *p<0.05, **p<0.01

Figure 11. Docking analysis visualization of VEGFR2 binding with complex 2

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