三甘醇桥联的愈创木薁二聚体的设计合成及抗炎、抗氧化活性研究

doi:10.6023/A25060209

摘要/Abstract

炎症介质释放失衡可诱发多种慢性疾病, 开发高效抗炎药物具有重要临床意义. 天然产物的桥联二聚体化可显著增强单体生物活性、选择性和稳定性. 本工作设计以天然双环倍半萜化合物愈创木薁(GA)为母体单元, 以三甘醇为连接片段, 合成了酯基和醚键连接的二聚体化合物(BGA-E和BGA-T), 并研究了新化合物与对比化合物(GA和愈创木薁磺酸钠)的抗炎和抗氧化活性. 通过1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除法和铁离子还原抗氧化能力(FRAP)法检测了化合物的抗氧化能力; 采用Griess法、常规酶联免疫分析(ELISA)法和Western blot实验检测化合物对炎症因子的抑制效果; 使用Cell Counting Kit-8 (CCK-8)法检测了化合物的细胞毒性. 结果显示, 化合物BGA-T能高效抑制脂多糖(LPS)诱导的炎症因子白细胞介素6 (IL-6)的释放, IL-6的分泌量降至空白对照组的34.89%, 抑制效果为GA的8倍. BGA-T还通过下调环氧合酶(COX-Ⅱ)的表达发挥抗炎效果. 以上化合物均无明显细胞毒性. 该研究为基于天然产物的桥联二聚体设计及抗炎药物的研发提供了新思路.

关键词: 愈创木薁, 三甘醇, 抗炎, 抗氧化, 天然产物

Imbalanced release of inflammatory mediators can trigger various chronic diseases, and the development of highly effective anti-inflammatory drugs holds significant clinical value. The bridged dimerization of natural products can markedly enhance the bioactivity, selectivity, and stability of monomers. This study designed and synthesized guaiazulene (GA)-based dimers (BGA-E and BGA-T) linked via ester and ether bonds, with triethylene glycol as the spacer and the natural bicyclic sesquiterpene GA as the core structure. The anti-inflammatory and antioxidant activities of the new compounds were investigated in comparison with the reference compounds (GA and sodium guaiazulene sulfonate). The antioxidant capacity of the compounds was determined using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay and the ferric ion reducing antioxidant power (FRAP) assay. The inhibitory effects of the compounds on inflammatory factors were evaluated using the Griess method, conventional enzyme-linked immunosorbent assay (ELISA), and Western Blot analysis. Cytotoxicity was measured via the Cell Counting Kit-8 (CCK-8) assay. The results demonstrated that compound BGA-T effectively suppressed the release of the inflammatory cytokine interleukin-6 (IL-6) induced by lipopolysaccharide (LPS), reducing IL-6 secretion to 34.89% of the blank control group. Its inhibitory effect was 8-fold stronger than GA. Additionally, BGA-T exerted its anti-inflammatory effects by downregulating the expression of cyclooxygenase-2 (COX-II). None of the tested compounds exhibited significant cytotoxicity. This study provides new insights for the design of bridged dimers based on natural products and the development of anti-inflammatory drugs.

Key words: guaiazulene, triethylene glycol, anti-inflammatory, antioxidant, natural products

引用此文

任蝶沨, 向焌钧, 高希珂. 三甘醇桥联的愈创木薁二聚体的设计合成及抗炎、抗氧化活性研究[J]. 化学学报, 2025, 83(10): 1174-1183.

Diefeng Ren, Junjun Xiang, Xike Gao. Design, Synthesis, and Anti-inflammatory/Antioxidant Activities of Guaiazulene Dimers Bridged by Triethylene Glycol[J]. Acta Chimica Sinica, 2025, 83(10): 1174-1183.

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图/表 15

图1. 青蒿素桥联二聚体的结构式

Figure 1. The structural formula of artemisinin-bridged dimers

图2. 愈创木薁的结构式

Figure 2. Chemical structure of Guaiazulene

图式1. BGA-E和BGA-T的合成路线

Scheme 1. Synthetic routes of BGA-E and BGA-T

图3. BGA-E和BGA-T的1H NMR谱图

Figure 3. 1H NMR spectra of compounds BGA-E and BGA-T

图4. 不同浓度的化合物分别在0.5 h (a)和4 h (b)时对DPPH自由基的清除率

Figure 4. The scavenging rates of different concentrations of compounds against DPPH radicals at 0.5 h (a) and 4 h (b) Data are given as mean±standard error of mean (SEM). Statistical analysis is performed by oneway analysis of variance (ANOVA). **p<0.01

化合物	时间/h	自由基清除率^a/%
化合物	时间/h	50 μmol•L⁻¹	100 μmol•L⁻¹	200 μmol•L⁻¹	400 μmol•L⁻¹
BGA-E	0.5	12.86	14.25	18.90	24.13
BGA-E	4	14.19	16.58	23.58	31.91
BGA-T	0.5	11.25	12.06	12.70	17.52
BGA-T	4	13.35	14.72	14.83	24.44
GA	0.5	11.84	11.86	13.88	10.15
GA	4	12.35	12.51	13.91	12.03
GA-SO₃Na	0.5	10.86	10.95	10.40	9.20
GA-SO₃Na	4	11.23	11.64	11.24	9.69

表1. 不同浓度的化合物在0.5 h和4 h时对DPPH自由基的清除率

Table 1. The scavenging rates of different concentrations of compounds against DPPH radicals at 0.5 h and 4 h

图5. FRAP法测定不同浓度化合物的OD值

Figure 5. Measurement of OD values for compounds at different concentrations by FRAP assay Data are given as mean±SEM

化合物	OD值
化合物	50 μmol•L⁻¹	100 μmol•L⁻¹	200 μmol•L⁻¹	400 μmol•L⁻¹
BGA-E	0.0685	0.0827	0.1011	0.1389
BGA-T	0.1005	0.1503	0.2267	0.3357
GA	0.1084	0.1772	0.3135	0.5741
GA-SO₃Na	0.0955	0.1370	0.2090	0.3346

表2. FRAP 法测定化合物的OD值

Table 2. Measurement of OD values for compounds at different concentrations by FRAP assay

图6. 不同浓度的化合物对LPS诱导的RAW 264.7细胞炎症模型的NO抑制率

Figure 6. NO inhibition rates of compounds at various concentrations in LPS-stimulated RAW 264.7 cells Data are given as mean±SEM. Statistical analysis is performed by Two-way analysis of variance (ANOVA). ****p<0.0001, ***p<0.001, **p<0.01 compared to GA and GA-SO3Na

化合物	浓度/(μmol•L⁻¹)	抑制率^a/%
BGA-E	10	16.60
	20	23.01
	40	36.23
	80	47.75
BGA-T	10	15.61
	20	28.57
	40	43.99
	80	57.92
GA	10	12.88
	20	19.52
	40	23.05
	80	27.19
GA-SO₃Na	10	4.49
	20	2.85
	40	6.18
	80	15.32
模型组	—	0

表3. 不同浓度的化合物对LPS诱导的RAW 264.7细胞炎症模型的NO抑制率

Table 3. NO inhibition rates of compounds at various concentrations in LPS-stimulated RAW 264.7 cells

图7. 不同化合物对LPS诱导的RAW 264.7细胞中TNF-α (a)和IL-6 (b)相对含量的影响

Figure 7. The effect of different compounds on the relative amount of TNF-α (a) and IL-6 (b) in LPS-induced RAW 264.7 cells Data are given as mean±SEM. Statistical analysis is performed by Two-way analysis of variance (ANOVA). ****p<0.0001, ***p<0.001, **p<0.01 compared to GA and GA-SO3Na

化合物	浓度/ (μmol•L⁻¹)	相对含量^a/%
化合物	浓度/ (μmol•L⁻¹)	TNF-α	IL-6
BGA-E	20	101.71	88.40
	40	105.33	71.76
	80	139.79	45.83
BGA-T	20	94.47	79.25
	40	89.67	63.89
	80	77.14	34.89
GA	20	101.29	98.72
	40	114.84	95.28
	80	122.68	92.17
GA-SO₃Na	20	88.96	103.13
	40	84.68	96.59
	80	82.98	91.48
模型组	—	100.00	100.00

表4. 化合物对LPS诱导的RAW 264.7细胞中TNF-α和IL-6的相对含量

Table 4. The relative amounts of TNF-α and IL-6 in LPS-induced RAW 264.7 cells with the compounds

图8. 不同化合物影响LPS诱导的RAW 264.7细胞中COX-II表达的WB条带图

Figure 8. Western blot bands showing the effects of different compounds on COX-II expression in LPS-induced RAW 264.7 cells

图9. 不同化合物对LPS诱导的RAW 264.7细胞中COX-II相对含量的影响

Figure 9. The effect of different compounds on the relative amount of COX-II in LPS-induced RAW 264.7 cells Data are given as mean±SEM. Statistical analysis is performed by One-way analysis of variance (ANOVA). ****p<0.0001, ***p<0.001, **p<0.01 compared to DMSO

图10. 化合物对巨噬细胞RAW 264.7细胞存活率的影响

Figure 10. Effects of compounds on the cell viability of macrophage RAW 264.7 cells

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