Anti-breast cancer sinomenine derivatives via mechanisms of apoptosis induction and metastasis reduction

Figures & data

Figure 1. The chemical structures of reported Sinomenine and furoxan derivatives.

Scheme 1. Synthesis of 1–4, 5A–E, 6A–E(a–c) and 7A–E(a–c). Reagents and conditions: (a) ClCH₂COOH, NaOH (aq), reflux, 2 h; (b) 30% H₂O₂, AcOH, rt, 3 h; (c) fuming HNO₃, 100 °C, 8 h; (d) corresponding diol, THF, 30% NaOH, 0 °C, 1 h; (e) corresponding anhydrides, TEA, DMAP, DCM, rt, 2 h; (f) DMAP, EDCI, rt, 4 h.

Table 1. The antiproliferative effects of the target compounds and parent compounds against different cell lines.

Compound	^aIC50 (μM)
	A549	MCF-7	MDA-MB-231	MCF-10A	^bSIMCF-7	^cSIMDA-MB-231
7Aa	15.25 ± 1.37	26.43 ± 1.25	31.51 ± 2.77	>50	>1.89	>1.58
7Ab	9.41 ± 0.68	15.31 ± 0.94	29.58 ± 1.80	>50	>3.27	>1.69
7Ac	2.33 ± 0.15	5.87 ± 0.42	1.62 ± 0.15	21.54 ± 0.88	3.67	13.30
7Ba	9.84 ± 0.47	18.96 ± 1.33	24.22 ± 1.97	>50	>2.64	>2.06
7Bb	4.91 ± 0.30	19.67 ± 1.28	16.45 ± 1.55	>50	>2.54	>3.04
7Bc	5.05 ± 0.21	12.18 ± 0.92	18.73 ± 0.61	>50	>4.11	>2.67
7Ca	2.97 ± 0.14	2.55 ± 0.14	2.26 ± 0.19	40.68 ± 2.79	15.95	18
7Cb	3.53 ± 0.29	4.68 ± 0.39	1.47 ± 0.14	36.71 ± 1.02	7.84	24.97
7Cc	1.94 ± 0.05	1.75 ± 0.15	0.82 ± 0.02	27.53 ± 2.34	15.73	33.57
7Da	2.79 ± 0.07	1.80 ± 0.11	0.92 ± 0.01	24.74 ± 1.67	13.74	26.89
7Db	3.92 ± 0.31	2.94 ± 0.27	3.11 ± 0.22	39.47 ± 2.90	13.43	12.69
7Dc	1.63 ± 0.05	2.18 ± 0.10	1.88 ± 0.09	19.85 ± 1.43	9.11	10.56
7Ea	4.62 ± 0.16	25.33 ± 0.90	19.00 ± 0.70	43.61 ± 2.65	2.79	2.30
7Eb	4.55 ± 0.43	17.89 ± 1.26	41.46 ± 1.48	>50	>2.79	>1.21
7Ec	4.81 ± 0.18	3.94 ± 0.28	1.81 ± 0.13	48.32 ± 3.78	12.26	26.70
Cisplatin	4.97 ± 0.37	^dNT	^dNT	^dNT	^eNC	^eNC
Adriamycin	^dNT	5.12 ± 0.31	4.92 ± 0.44	^dNT	^eNC	^eNC

^aIC₅₀: half inhibitory concentrations measured by the CCK-8 assay. The values are expressed as average ± standard deviation of three independent experiments.

^bSI_MCF-7: selectivity index between MCF-7 and MCF-10A. It was calculated as: SI = IC_50(MCF-10A)/IC_50(MCF-7).

^cSI_MDA-MB-231: selectivity index between MCF-7 and MDA-MB-231. It was calculated as: SI = IC_50(MCF-10A)/IC_{50(MDA-MB-231)}.

^dNT: not tested.

^eNC: not calculated.

Figure 2. NO-releasing ability of several selected target compounds. The values were expressed as averages of three independent experiments.

Figure 3. MDA-MB-231 cells were treated with 7Cc and Cells were stained with PI and investigated by flow cytometry. Data are represented as mean ± SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs control group. Statistical analyses were carried out on GraphPad.Prism software under ordinary one-way ANOVA method and compared with the control group.

Figure 4. Flow cytometry analysis of apoptosis induced by 7Cc in MDA-MB-231 cells. Data are represented as mean ± SD of three independent experiments. **p < 0.01, ***p < 0.001 vs control group. Statistical analyses were carried out on GraphPad.Prism software under ordinary one-way ANOVA method and compared with the control group.

Figure 5. 7Cc induced mitochondrial depolarisation in MDA-MB-231 cells. Data are represented as mean ± SD of three independent experiments. **p < 0.01, ***p < 0.001 vs control group. Statistical analyses were carried out on GraphPad.Prism software under ordinary one-way ANOVA method and compared with the control group.

Figure 6. (a) MDA-MB-231 cells were treated with 7Cc, sterile pipette tips were used to scratch evenly, the incubation was continued, and representative images were captured. (b) MDA-MB-231 cells were seeded onto chambers and incubated with 7Cc, stained with crystal violet, and representative images were photographed. (c) MDA-MB-231 cells were incubated with 7Cc, then fixed, washed and photographed with a fluorescence microscope. All data are represented as the mean ± SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs the control group. Statistical analyses were carried out on GraphPad.Prism software under ordinary one-way ANOVA method and compared with a control group.

Figure 7. MDA-MB-231 cells were incubated with 7Cc. Comet assay was used to evaluate DNA damage and photomicrographs were provided. Data are represented as mean ± SD of three independent experiments. *p < 0.05, **p < 0.01 vs control group. Statistical analyses were carried out on GraphPad.Prism software under ordinary one-way ANOVA method and compared with a control group.