杀菌剂氟苯醚酰胺的创制

曾令强; 罗睿童; 陈琼; 郝格非; 朱晓磊; 杨光富

doi:10.16801/j.issn.1008-7303.2022.0028

杀菌剂氟苯醚酰胺的创制

doi: 10.16801/j.issn.1008-7303.2022.0028

1.
华中师范大学化学学院农药与化学生物学教育部重点实验室，武汉 430079

基金项目: 湖北省科技厅重点研发项目 (2020BBA052) ；国家自然科学基金面上项目 (21977035，21837001).

详细信息

作者简介:
曾令强，zenglingqiang@mails.ccnu.edu.cn

罗睿童，ruitong_luo@163.com

通讯作者:
朱晓磊，xlzhu@mail.ccnu.edu.cn

杨光富，gfyang@mail.ccnu.edu.cn

中图分类号: TP391；TQ450.1
计量
- 文章访问数: 145
- HTML全文浏览量: 49
- PDF下载量: 28
- 被引次数: 0
出版历程
- 收稿日期: 2021-11-08
- 录用日期: 2022-01-05
- 网络出版日期: 2022-03-24

Discovery of fungicide flubeneteram

1.
Key Laboratory of Pesticide & Chemical Biology, Ministry of Education, College of Chemistry, Central China Normal University, Wuhan 430079, China

Funds: the Key Research and Development Program of Hubei Province (2020BBA052) and the National Natural Science Foundation of China (21977035, 21837001).

摘要

摘要: 琥珀酸脱氢酶 (succinate dehydrogenase, SDH) 是重要的杀菌剂靶标之一，而很多植物病原菌对靶向SDH的杀菌剂已经产生了较为严重的抗药性，因此新型靶向SDH的杀菌剂设计显得尤为重要。基于药效团连接碎片的虚拟筛选 (PFVS) 是一种独立于生物物理技术的高通量药物发现方法，采用PFVS方法成功获得了靶向SDH的新型杀菌剂候选化合物—氟苯醚酰胺。本文主要从PFVS原理、先导化合物的发现、取代基的修饰以及杀菌活性研究等方面对氟苯醚酰胺的创制进行系统分析。氟苯醚酰胺创制的案例分析可为农药研究工作者提供新思路和新方法。
- 药效团连接碎片的虚拟筛选 /
- 琥珀酸脱氢酶 /
- 氟苯醚酰胺 /
- 杀菌剂
Abstract: Succinate dehydrogenase (SDH) is one of the important fungicide targets. However, many plant pathogens showed medium and even high resistance to commercial fungicides targeting upon SDH. So, it is very urgent to design new novel inhibitor for SDH. Pharmacophore-linked fragment virtual screening (PFVS) is a high-throughput drug discovery approach independent of biophysical screening techniques and flubeneteram, a novel fungicide candidate targeting SDH, was successfully obtained by PFVS. Here, the development process of the fungicide flubeneteram was analyzed in detail, including the principle of PFVS, the discovery of lead compounds, the modification of substituents and the study on fungicidal activity. The discovery process of flubeneteram would provide novel ideas and methods for pesticide researchers.
- pharmacophore-linked fragment virtual screening(PFVS) /
- succinate dehydrogenase /
- flubeneteram /
- fungicide

HTML全文

1 氟苯醚酰胺 (试验代号：Y13149，本文中化合物11) 的化学结构式

1. The chemical structure of flubeneteram (also named Y13149, compound 11)

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图 1 药效团连接碎片虚拟筛选工作流程^[23]

Figure 1. Work flow of PFVS^[23]

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2 10个商品化SDHI的化学结构式(红色表示酸部分，蓝色表示胺部分)^[24]

2. The chemical structures of ten commercial SDHI (The acid and the amine parts were colored by red and blue, respectively)^[24]

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图 2 10个商品化SDHI与SDH作用方式叠合显示图 (sub_amine为胺部分，sub_acid为酸部分，HEM为血红素)^[24]

Figure 2. The overlay of the binding model of ten SDHI with SDH (sub_amine means the amine part, sub_acid means the acid part and HEM stands for heme)^[24]

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3 基于PFVS筛选得到的8个hit化合物 (蓝色表示PFVS确定的部分)^[25]

3. Chemical structures of hit compounds identified by PFVS (The fragments determined by PFVS are shown in blue)^[25]

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图 3 结合模式显示图

注：(A) hit4 (黄色棍状显示)与猪心SDH的结合方式； (B) 氟苯醚酰胺 (黄色棍状显示)与猪心SDH的结合方式； (C) 氟酰胺衍生物 (黄色棍状显示) 与猪心SDH结合方式 (PDB ID：3ABV)； (D) 氟苯醚酰胺 (橙色棍状显示)与水稻纹枯病菌SDH的结合方式； (E) 氟苯醚酰胺与猪心SDH (玫红色和黄色棍状显示) 和水稻纹枯病菌SDH (灰色和绿色棍状显示) 结合方式显示叠合图； (F) 噻呋酰胺与猪心SDH (玫红色和黄色棍状显示) 和水稻纹枯病菌SDH (灰色和绿色棍状显示) 结合方式显示叠合图；为了便于展示，C_M39/W的含义为猪心中SDH为C_M39，而水稻纹枯病中SDH为C_W39；以此类推，C_W35/P表示的是猪心中SDH为C_W35，而水稻纹枯病中SDH为C_P35；C_I30/F表示的是猪心中SDH为C_I30，而水稻纹枯病中SDH为C_F30^[26]。

Figure 3. The binding modes

Note: (A) hit 4 (yellow stick) with porcine SDH. (B) flubeneteram (yellow stick) with porcine SDH. (C) flutolanil derivatives (yellow stick) with porcine SDH in crystal structure 3ABV. (D) flubeneteram (orange stick) with Rhizoctonia solani (R. solani) SDH. (E) Overlay of flubeneteram binding with porcine SDH (represented by magenta and yellow stick) and R. solani SDH (represented by gray and green stick). (F) Overlay of thifluzamide binding with porcine SDH (represented by magenta and yellow) and R. solani SDH (represented by gray and green sticks). C_M39/W, C_W35/P, and C_I30/F respectively indicate C_M39, C_W35, and C_I30 in porcine SDH and C_W39, C_P35, and C_F30 in R. solani SDH. For clarity, just some key residues are shown^[26]

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图 4 化合物设计思路

注：^a指200 mg/L下该化合物对水稻纹枯病的防治效果。

Figure 4. Compound design strategy

Note: ^a Fungicidal activity against R. solani at a concentration of 200 mg/L.

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4 化合物11合成路线^[26]

4. Synthetic route of compound 11^[26]

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表 1 商品化SDHI与SDH的结合自由能 (kcal/mol)^c^[24]

Table 1. The binding free energies of ten SDHI with the Q-site of SDH (kcal/mol)^c^[24]

化合物 Compound	△E_ele	△E_vdw	△G_np	△G_pol	△H	−T△S	△G_cal	△G_exp^a	K_i^b/(μmol/L)
噻呋酰胺 thifluzamide	−10.62	−43.99	−4.76	31.02	−28.35	11.21	−17.14	−9.10	0.20
吡噻菌胺 penthiopyrad	−13.84	−41.91	−5.10	34.28	−26.57	13.54	−13.03	−7.35	3.90
萎锈灵 carboxin	−14.42	−35.65	−4.15	29.48	−24.74	12.16	−12.58	−7.25	4.59
啶酰菌胺 boscalid	−18.50	−39.68	−4.66	39.76	−23.08	11.02	−12.06	−6.84	9.20
麦锈灵 benodanil	−10.38	−34.43	−4.20	27.87	−21.14	10.13	−11.01	−6.63	13.17
氟酰胺 flutolanil	−9.75	−38.41	−4.68	32.28	−20.57	9.76	−10.81	−6.17	28.88
呋吡菌胺 furamepyr	−13.94	−37.16	−4.58	34.69	−20.99	10.43	−10.56	−6.16	29.27
灭锈胺 mepronil	−11.92	−36.64	−4.57	32.97	−20.16	9.59	−10.57	−6.11	31.81
氧化萎锈灵 oxycarboxin	−20.09	−38.22	−4.26	41.65	−20.93	10.48	−10.45	−5.56	81.02
甲呋酰胺 fenfuram	−15.03	−32.00	−3.90	29.13	−21.79	11.64	−10.15	−5.39	108.40
注：^a△G_exp = -RTlnK_i；^b使用琥珀酸-DCIP体系测定。^c1 kcal/mol = 4.184 kJ/mol。 Note：^a△G_exp = -RTlnK_i; ^bDetermined with the succinate-DCIP system. ^c1kcal/mol = 4.184 kJ/mol.

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表 2 活体杀菌活性及对猪心SDH的IC₅₀值^[26]

Table 2. Fungicidal activities in vivo and IC₅₀ value against porcine SDH^[26]


化合物 Compound	R¹	R²	质量浓度 Concentration/(mg/L)	防治效果 Control efficacy/%		IC₅₀值 (猪心SDH) IC₅₀ value (porcine SDH)/(μmol/L)
化合物 Compound	R¹	R²	质量浓度 Concentration/(mg/L)	水稻纹枯病 Rice sheath blight	黄瓜白粉病 Cucumber powdery mildew	IC₅₀值 (猪心SDH) IC₅₀ value (porcine SDH)/(μmol/L)
Hit4	CF₃	H	200	76	100	19.79
2	CF₃	2-Cl	200	82	56	1.98
3	CF₃	3-Cl	200	69	22	36.9
4	CF₃	4-F	200	57	15	>100
5	CF₃	4-Cl	200	66	44	4.90
6	CF₃	4-CN	200	30	0	>100
7	CF₃	2,4-Cl₂	200	69	0	2.70
8	CF₃	2-Cl-4-CF₃	200	100	100	0.28
9	CHF₂	2-Cl	200	100	100	1.13
10	CHF₂	2,4-Cl₂	200	100	80	0.40
11	CHF₂	2-Cl-4-CF₃	200	100	100	0.11
吡噻菌胺 penthiopyrad	—	—	NT	NT	NT	1.29
噻呋酰胺 thifluzamide	—	—	100	100	100	NT
注：NT表示未测定。Note: NT indicates untested.

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表 3 化合物9~11在低浓度下活体杀菌活性^[26]

Table 3. Fungicidal activities of compounds 9-11 in vivo at lower concentration^[26]

化合物 Compound	质量浓度 Concentration/ (mg/L)	防治效果 Control efficacy/%		化合物 Compound	质量浓度 Concentration/ (mg/L)	防治效果 Control efficacy/%
化合物 Compound	质量浓度 Concentration/ (mg/L)	水稻纹枯病 Rice sheath blight	黄瓜白粉病 Cucumber powdery mildew	化合物 Compound	质量浓度 Concentration/ (mg/L)	水稻纹枯病 Rice sheath blight	黄瓜白粉病 Cucumber powdery mildew
9	100	100	100	11	100	100	100
	50	100	83		50	100	100
	25	80	41		25	99	94
	12.5	72	7		12.5	95	67
	6.25	75	0		6.25	70	0
10	100	100	—	噻呋酰胺 thifluzamide	100	100	100
	50	98	—		50	100	63
	25	95	—		25	82	22
	12.5	82	—		12.5	61	0
	6.25	69	—		6.25	54	0
注：— 表示未测定。Note: — Indicates untested.

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表 4 化合物11对水稻纹枯病的田间药效试验^[26,28]

Table 4. Control efficacy of compound 11 against R. solani in field trials ^[26,28]

	药剂处理 Treatment	有效成分用量 Amount of active ingredient/(g/hm²)	第 2 次施药后 7 d 7 days after the second spraying		第 2 次施药后 21 d 21 days after the second spraying
	药剂处理 Treatment	有效成分用量 Amount of active ingredient/(g/hm²)	病情指数 Disease index	防治效果 Control efficacy/%	病情指数 Disease index	防治效果 Control efficacy/%
浙江宁波 Ningbo, Zhejiang	11 (5%, EC)	112.5	1.59	77.2	0.99	88.5
		75.0	2.12	69.6	1.62	81.2
		37.5	2.40	65.5	1.87	78.3
	噻呋酰胺 thifluzamide (24%, SC)	75.0	3.59	48.4	2.12	75.4
	清水 Water	0	6.97	0	8.63	0
浙江绍兴 Shaoxing, Zhejiang	11 (5%, EC)	112.5	2.22	83.4	6.70	83.6
		75.0	2.78	78.8	9.00	78.0
		37.5	3.70	72.9	11.9	70.6
	噻呋酰胺 thifluzamide (24%, SC)	75.0	2.63	80.8	8.52	79.1
	清水 Water	0	6.97	0.00	8.63	0.00

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