新型1,2,3-三氮唑类衍生物的合成及抗炎活性研究

doi:10.6023/cjoc202209033

摘要/Abstract

吲哚胺2,3-双加氧酶2 (IDO2)在炎症特别是类风湿性关节炎(RA)方面的作用逐渐被阐明, IDO2成为RA治疗的潜在靶点. 目前尚未见活性强、选择性高的IDO2小分子抑制剂被报道. 以吲哚胺2,3-双加氧酶1 (IDO1)抑制剂epacadostat为先导化合物, 借助计算机辅助药物设计, 设计合成了24个未见文献报道的1,2,3-三氮唑类衍生物. 生物活性研究表明, 目标化合物均具有IDO1和IDO2酶抑制活性, 其中(Z)-N-((1-(2-((4-(N-(3-溴-4-氟苯基)-N'-羟基甲脒)-1,2,5-噁二唑-3-基)氨基)乙基)-1H-1,2,3-三氮唑-4-基)甲基)呋喃-2-甲酰胺(I-11)在100 nmol/L浓度下对IDO1和IDO2酶的抑制率分别为76%和49%, 优于先导化合物epacadostat(分别为62%和25%). 抗炎活性研究表明, 化合物I-11可有效抑制脂多糖(LPS)诱导的RAW264.7细胞炎症因子TNF-α的分泌, 并且对小鼠耳肿胀抑制率(56.81%, 100 mg/kg, 灌胃)优于阳性对照Naproxen (45.29%, 150 mg/kg, 灌胃), 表现出较好的抗炎活性.

关键词: 吲哚胺2,3-双加氧酶1 (IDO1), 吲哚胺2,3-双加氧酶2 (IDO2), 1,2,3-三氮唑, 抗炎活性

As the role of indoleamine 2,3-dioxygenase 2 (IDO2) in inflammation, especially in rheumatoid arthritis (RA), is gradually being explained, IDO2 has become a potential target for the treatment of RA. No small molecule inhibitor of IDO2 with strong activity and high selectivity has been reported yet. With the help of computer-aided drug design, twenty-four 1,2,3-triazole derivatives were designed and synthesized by using epacadostat as a lead compound. The results of biological activity show that all target compounds display the inhibitory effects on IDO1 and IDO2 enzymes. Among them, the inhibition rates of (Z)-N-((1-(2-((4-(N-(3-bromo-4-fluorophenyl)-N'-hydroxyformamidine)-1,2,5-oxadiazo-3-yl)amino)ethyl)-1H-1,2,3- triazol-4-yl)methyl) (I-11) on indoleamine 2,3-dioxygenase 1 (IDO1) and IDO2 enzymes were 76% and 49% at 100 nmol/L concentration, respectively, which are better than those of epacadostat (62% and 25%). The anti-inflammatory activity study showed that compound I-11 could significantly inhibit the secretion of inflammatory factor TNF-α at the cellular level. Compound I-11 also showed good in vivo anti-inflammatory activity in mouse ear swelling model, and the inhibition rate was 56.81% at the dose of 100 mg/kg (Ig) which was better than that of naproxen (45.29%, 150 mg/kg, Ig).

Key words: indoleamine 2,3-dioxygenase 1 (IDO1), indoleamine 2,3-dioxygenase 2 (IDO2), 1,2,3-triazole, anti-inflammatory activity

引用此文

孙泽人, 翟冰新, 何光超, 沈慧, 陈琳雅, 张杉, 邹毅, 朱启华, 徐云根. 新型1,2,3-三氮唑类衍生物的合成及抗炎活性研究[J]. 有机化学, 2023, 43(6): 2143-2155.

Zeren Sun, Bingxin Zhai, Guangchao He, Hui Shen, Linya Chen, Shan Zhang, Yi Zou, Qihua Zhu, Yungen Xu. Synthesis and Anti-inflammatory Evaluation of Novel 1,2,3-Triazole Derivatives[J]. Chinese Journal of Organic Chemistry, 2023, 43(6): 2143-2155.

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图/表 9

图1. IDO1-epacadostat共晶结构(绿色)与IDO2-epacadostat对接结果(红色)叠合图

Figure 1. Overlay of IDO1-epacadostat co-crystal structure (green) and IDO2-epacadostat docking results (red)

图式1. 目标化合物的设计思想

Scheme 1. Design strategy of the target compounds

图式2. 目标化合物的合成路线

Scheme 2. Synthetic routes to target compounds

Compd.	Inhibition rate/%
Compd.	IDO1 (100 nmol/L)	IDO2 (100 nmol/L)	IDO2 (1 μmol/L)
I-1	40	27	72
I-2	52	12	48
I-3	68	14	47
I-4	67	18	63
I-5	59	26	60
I-6	70	34	65
I-7	75	30	49
I-8	37	14	32
I-9	65	18	51
I-10	65	10	54
I-11	76	49	96
I-12	77	12	76
I-13	66	18	87
I-14	61	20	50
I-15	62	10	40
I-16	64	10	45
I-17	57	16	42
I-18	42	9	37
I-19	49	15	61
I-20	67	5	41
I-21	36	12	59
I-22	15	4	36
I-23	43	4	44
I-24	44	3	32
Epacadostat	62	25	—^a

表1. 化合物I-1~I-24对IDO1和IDO2酶的体外抑制活性

Table 1. Inhibitory activity of compounds I-1~I-24 against IDO1 and IDO2 in vitro

Compd.	IC₅₀/(μmol•L^–1)
I-7	40
I-11	37
Epacadostat	74

表2. 化合物I-7和I-11对正常大鼠肾细胞的体外抗增殖活性

Table 2. In vitro antiproliferative activity of compounds I-7 and I-11 against NRK cells

Compd.	Cell viability/%				IC₅₀/(μmol•L^–1)
Compd.	50 μmol/L	16.67 μmol/L	5.55 μmol/L	1.85 μmol/L	IC₅₀/(μmol•L^–1)
I-11	32.8	74.4	78.3	76.9	57
I-7	9.1	55.6	59.2	65.3	>16
Naproxen	35.5	>100	>100	>100	—^a
Epacadostat	40.5	>100	>100	>100	—
I-11+LPS (1 μg/mL)	53.8	74.5	80.6	91.2	69
I-7+LPS (1 μg/mL)	47.5	91.5	>100	>100	48
Naproxen+LPS (1 μg/mL)	44.4	>100	>100	>100	—
Epacadostat+LPS (1 μg/mL)	44.4	>100	>100	>100	—

表3. 化合物I-7和I-11对RAW264.7细胞活力的影响及IC50

Table 3. Effects of compounds I-7 and I-11 on RAW264.7 cell viability and IC50

图2. 化合物对LPS诱导的RAW264.7细胞分泌TNF-α的影响

Figure 2. Effects of compounds on LPS-induced RAW264.7 cells secretion of TNF-α Error bars represent s.d., **P<0.01, ***P<0.001 vs. control group.

Group	Swelling rate/%	Swelling inhibition ratio/%
Model	58.21±12.79	—
Naproxen (150 mg/kg)	31.85±17.29	45.29
I-11 (100 mg/kg)	25.14±15.80	56.81

表4. 化合物I-11对二甲苯致小鼠耳肿胀的影响

Table 4. Effects of compound I-11 on xylene-induced ear swelling in mice (n=8)

图3. (A)化合物I-11与IDO1蛋白对接结果以及(B)化合物I-11与IDO2蛋白对接结果

Figure 3. (A) The docking result of compound I-11 and IDO1 protein, and (B) the docking result of compound I-11 and IDO2 protein

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