曲酸及氯曲酸衍生物的合成及抑菌活性

引用本文

张文豪, 魏少鹏, 姬志勤. 曲酸及氯曲酸衍生物的合成及抑菌活性[J]. 农药学学报,2014,16(1): 92-96 复制到剪切板

ZHANG Wenhao, WEI Shaopeng, JI Zhiqin. Synthesis and antifungal activity of kojic acid and chlorokojic acid derivatives[J]. Chinese Journal of Animal Nutrition, 2014,16(1): 92-96 复制到剪切板

曲酸及氯曲酸衍生物的合成及抑菌活性

张文豪 , 魏少鹏, 姬志勤

西北农林科技大学植物保护学院, 陕西杨凌 712100

收稿日期:2013-12-4;修回日期: 2013-12-18 .

基金项目：国家自然科学基金（31371973，31301700）；陕西省自然科学基金（2013JQ3003）；中央高校基本科研业务费专项（QN2013009）.

作者简介：张文豪，男，硕士研究生，E-mail：wenhao294@qq.com

通讯作者：姬志勤，男，副研究员，主要从事天然产物化学研究，E-mail：jizhiqin@nwsuaf.edu.cn

摘要：以曲酸为原料，设计合成了16个曲酸衍生物（QⅠ-01~QⅠ-16）和10个氯曲酸衍生物（QⅡ-01~QⅡ-10），通过核磁共振氢谱及元素分析对其结构进行了表征。初步抑菌活性测定结果表明：所有供试化合物对苹果炭疽病菌Glomerella cingulata和番茄灰霉病菌Botrytis cinerea的菌丝生长均有不同程度的抑制作用，其中化合物 QⅠ-07 和 QⅠ-11 对苹果炭疽病菌活性最高，50 μg/mL时的抑制率分别为79.3%和86.2%；化合物 QⅡ-04 、 QⅡ-08和QⅡ-09 对番茄灰霉病菌活性最高，50 μg/mL时的抑制率分别为70.0%、70.0%和76.7%。

关键词：曲酸氯曲酸酰化合成抑菌活性

Synthesis and antifungal activity of kojic acid and chlorokojic acid derivatives

ZHANG Wenhao , WEI Shaopeng, JI Zhiqin

College of Plant Protection, Northwest A & F University, Yangling 712100, Shaanxi Province, China

Abstract: Sixteen kojic acid derivatives (QⅠ-01~QⅠ-16) and ten chlorokojic acid derivatives (QⅡ-01~QⅡ-10) were synthesized from kojic acid. The structures of the synthesized derivatives were confirmed by ¹H NMR and elemental analysis. Antifungal activity results indicated that all of the title compounds exhibited variable inhibitory activities against the mycelial growth of Glomerella cingulata and Botrytis cinerea. The inhibition rates of 50 μg/mL QⅠ-07 and QⅠ-11 against G.cingulata were 79.3% and 86.2%, and QⅡ-04, QⅡ-08 and QⅡ-09 against B.cinerea were 70.0%, 70.0% and 76.6%, respectively.

Key words: kojic acid chlorokojic acid acylation synthesis antifungal activity

曲酸(kojic acid)，化学名称为5-羟基-2-羟甲基-1,4-吡喃酮，是曲霉属和青霉属多种真菌好氧发酵产生的弱酸性次生代谢物^[1]，具有多种生物活性，包括抗细菌^[2,3]、抗真菌^[4,5]、抗病毒^[6]、抑制酪氨酸酶及抗氧化活性^[7]等。此外，也有文献报道曲酸对小菜蛾酚氧化酶具有明显的抑制作用，是酚氧化酶典型的竞争抑制剂^[8,9]。在从植物内生菌中进行抗生素筛选时，笔者所在课题组曾从豇豆中分离得到1株Aspergillus flavus F52菌株，并从该菌株的发酵产物中获得了一种高产量的化合物，经与相关文献的核磁共振和红外光谱比对，鉴定其为曲酸^[10]。鉴于曲酸具有多种生物活性，笔者以曲酸为原料，设计合成了16个曲酸衍生物和10个氯曲酸衍生物，其中21个为未见文献报道的新化合物，其合成路线见Scheme 1。采用抑制菌丝生长速率法初步评价了目标化合物对苹果炭疽病菌Glomerella cingulata和番茄灰霉病菌Botrytis cinerea的抑菌活性。

Scheme 1

1 实验部分

1.1 仪器与试剂

WPR熔点仪(温度未校正，上海精科仪器有限公司)；Bruker Avance III 核磁共振仪(瑞士Bruker BioSpin公司,以TMS为内标，CDCl₃为溶剂)；Vairo EL型元素分析仪(德国Elementar元素分析系统公司)。

二氯甲烷(DCM)经氯化钙干燥，三乙胺(Et₃N)经氢氧化钾干燥，其余试剂均为市售分析纯；柱层析硅胶(48～75 μm)，青岛海洋化工厂生产；对照药剂嘧菌酯(azoxystrobin)原药，纯度98%，上虞颖泰精细化工有限公司生产。

1.2 化合物的合成

1.2.1 曲酸衍生物(QI-01～QI-16)的合成通法

以化合物QI-01为例，在干燥的100 mL梨形瓶中，依次加入142 mg(1 mmoL)曲酸和5 mL二氯甲烷，搅拌溶解后加入200 μL三乙胺，0～5 ℃下缓慢滴加乙酰氯(0.078 g，1 mmol，溶于适量二氯甲烷)溶液。滴加完毕待体系缓慢恢复至室温后继续反应2 h，TLC [V(石油醚)∶V(乙酸乙酯)=4∶1] 跟踪至原料反应完毕。依次用20 mL饱和碳酸氢钠、水和饱和氯化钠水溶液洗涤，无水硫酸钠干燥。硅胶柱层析 [V(石油醚)∶V(乙酸乙酯)=6∶1] 纯化得无色油状物0.174 g，收率77%。按照相同的方法合成了其余化合物QI-02～QI-16。

1.2.2 氯曲酸衍生物(QII-01～QII-10)的合成通法

参考文献^[6]方法，先由曲酸与亚硫酰氯(SOCl₂)在冰浴下反应生成氯曲酸，再参照1.2.1节方法合成化合物QII-01～QII-10。

1.3 抑菌活性测定

供试苹果炭疽病菌Glomerella cingulata和番茄灰霉病菌Botrytis cinerea，由西北农林科技大学农药研究所提供。采用平皿法^[11]分别测定目标化合物在 50 μg/mL 下对苹果炭疽病菌和番茄灰霉病菌菌丝生长的抑制作用，以10 μg/mL的嘧菌酯为对照药剂。称取1～2 mg待测化合物，用二甲亚砜溶解并配制成质量浓度为10 mg/mL的母液，备用。

2 结果与讨论

2.1 化合物的合成

以曲酸为起始原料，经酰化反应获得16个曲酸衍生物和10个氯曲酸衍生物，通过 ¹H NMR和元素分析对其结构进行了表征，其中21个为新化合物。其理化数据和元素分析数据见表1，核磁数据见表2。

表1 目标化合物的理化数据和元素分析数据 Table 1 Physico-chemical and elemental analysis data of target compounds

化合物

Compd.

熔点

m.p./℃

收率

Yield/%

分子式

Formula

元素分析(计算值) Elemental analysis (Calcd.)/%

QI-01^*	CH₃CO	99.5～101.9(100～102^[12])	77	C₁₀H₁₀O₆	53.10(53.11)	4.46 (4.47)
QI-02	CHCl₂CO	—	69	C₁₀H₆Cl₄O₆	33.00 (33.08)	1.66 (1.62)
QI-03	CH₂ClCO	—	64	C₁₀H₈Cl₂O₆	40.70 (40.68)	2.73 (2.69)
QI-04	CH₃(CH₂)₂CH₂CO	—	51	C₁₆H₂₂O₆	61.92 (61.91)	7.15 (7.17)
QI-05^*	C(CH₃)₃CO	101.8～102.5(102.5^[13])	73	C₁₆H₂₂O₆	61.92 (61.90)	7.15 (7.19)
QI-06	CH₂=C(CH₃)CO	—	81	C₁₄H₁₄O₆	60.43 (60.44)	5.07 (5.10)
QI-07	CH₃CH₂C(C₆H₅)HCO	—	82	C₂₆H₂₆O₆	71.87 (71.88)	6.03 (6.00)
QI-08^*	CH₃(CH₂)₅CH₂CO	57.6～59.5(58～60^[14])	75(83^[14])	C₂₂H₃₄O₆	66.98 (67.00)	8.69 (8.67)
QI-09	CH(CH₃)₂CH₂OCO	—	52	C₁₆H₂₂O₈	56.13 (56.11)	6.48 (6.51)
QI-10^*	C₆H₅CO	132.5～134.3(133～134^[15])	61(79^[15])	C₂₀H₁₄O₆	68.57 (68.55)	4.03 (3.99)
QI-11	o-CH₃C₆H₄CO	100.0～101.8	65	C₂₂H₁₈O₆	69.83 (69.85)	4.79 (4.81)
QI-12	m-CH₃C₆H₄CO	77.3～79.1	55	C₂₂H₁₈O₆	69.83 (69.78)	4.79 (4.75)
QI-13	o-ClC₆H₄CO	83.7～85.2	52	C₂₀H₁₂Cl₂O₆	57.30 (57.28)	2.89 (2.88)
QI-14	m-ClC₆H₄CO	100.4～102.1	57	C₂₀H₁₂Cl₂O₆	57.30 (57.28)	2.89 (2.91)
QI-15	p-ClC₆H₄CO	140.8～142.1	67	C₂₀H₁₂Cl₂O₆	57.30 (57.34)	2.89 (2.88)
QI-16	p-F₃CC₆H₄CO	163.8～165.1	71	C₂₂H₁₂F₆O₆	54.33 (54.35)	2.49 (2.51)
QII-01^*	CH₃CO	95.7～96.8(96～97^[16])	75(94^[16])	C₈H₇ClO₄	47.43 (47.41)	3.48 (3.52)
QII-02	cyclo-C₃H₅CO	74.8～76.5	68	C₁₀H₉ClO₄	52.53 (52.55)	3.97 (4.01)
QII-03	CH₃CH₂CH₂CO	—	66	C₁₀H₁₁ClO₄	52.07 (52.02)	4.81 (4.79)
QII-04	CH₃(CH₂)₂CH₂CO	—	70	C₁₁H₁₃ClO₄	54.00 (54.03)	5.36 (5.32)
QII-05	C(CH₃)₃CO	106.4-107.8	77	C₁₁H₁₃ClO₄	54.00 (54.01)	5.36 (5.37)
QII-06	CH(CH₃)₂CH₂OCO	—	69	C₁₁H₁₃ClO₅	50.68 (50.70)	5.03 (5.00)
QII-07	CH₃(CH₂)₅CH₂CO	—	71	C₁₄H₁₉ClO₄	58.64 (58.66)	6.68 (6.71)
QII-08	m-CH₃C₆H₄CO	72.2～73.4	68	C₁₄H₁₁ClO₄	60.34 (60.32)	6.98 (7.01)
QII-09	CH₃CH₂C(C₆H₅)HCO	—	71	C₁₆H₁₅ClO₄	62.65 (62.64)	4.93 (4.97)
QII-10	p-F₃CC₆H₄CO	108.1～109.4	75	C₁₄H₈ClF₃O₄	50.55 (50.51)	2.42 (2.39)

注：“^*”，已知化合物；“—”, 油状或蜡状物，熔点未测定。

Note：“^*”，known compounds. “—”, the oil or waxy compound, melting point not tested.

表1 目标化合物的理化数据和元素分析数据

Table 1 Physico-chemical and elemental analysis data of target compounds

表2 目标化合物的 ¹H NMR数据(CDCl₃, 500 MHz)

Table 2 ¹H NMR data of target compounds (CDCl₃, 500 MHz)

表2(Continued) 目标化合物的 ¹H NMR数据(CDCl₃, 500 MHz) Table 2(Continued) ¹H NMR data of target compounds (CDCl₃, 500 MHz)

化合物Compd.	¹H NMR， δ_H
QI-09	0.97 (d, J=8.0 Hz, 6H, 2×CH₃), 0.98 (d, J=8.0 Hz, 6H, 2×CH₃), 1.97～2.08 (m, 2H, 2×CH), 3.99 (d, J=6.5 Hz, 2H, CH₂), 4.01 (d, J=6.5 Hz, 2H, CH₂), 4.98 (s, 2H, CH₂), 6.55 (s, 1H, CH), 7.99 (s, 1H, CH)
QI-11	2.63 (s, 3H, CH₃), 2.65 (s, 3H, CH₃), 5.17 (s, 2H, CH₂), 6.65 (s, 1H, CH), 7.28～7.32 (m, 4H, Ar-H), 7.44～7.48 (m, 2H, Ar-H), 8.00 (d, J=8.0 Hz, 1H, Ar-H), 8.03 (s, 1H, CH), 8.16 (d, J=8.0 Hz, 1H, Ar-H)
QI-12	2.42 (s, 3H, CH₃), 2.43 (s, 3H, CH₃), 5.20 (s, 2H, CH₂), 6.66 (s, 1H, CH), 7.35～7.45 (m, 4H, Ar-H), 7.89 (d, J=8.0 Hz, 2H, Ar-H), 7.99 (d, J=8.0 Hz, 2H, Ar-H), 8.04 (s, 1H, CH)
QI-13	5.21 (s, 2H, CH₂), 6.69 (s, 1H, CH), 7.36～7.39 (m, 2H, Ar-H), 7.48～7.51 (m, 4H, Ar-H), 7.92 (d, J=8.0 Hz, 1H, Ar-H), 8.08 (s, 1H, CH), 8.14 (d, J=8.0 Hz, 1H, Ar-H)
QI-14	5.22 (s, 2H, CH₂), 6.67 (s, 1H, CH), 7.42～7.46 (m, 2H, Ar-H), 7.59～7.63 (m, 2H, Ar-H), 7.97～7.99 (m, 1H, Ar-H), 8.04～8.06 (m, 3H, CH, Ar-H), 8.14～8.15 (m, 1H, Ar-H)
QI-15	5.21 (s, 2H, CH₂), 6.65 (s, 1H, CH), 7.46 (d, J=1.5 Hz, 2H, Ar-H), 7.48 (d, J=1.5 Hz, 2H, Ar-H), 8.01 (s, 1H, Ar-H), 8.04 (s, 1H, Ar-H), 8.06 (s, 1H, CH), 8.09 (s, 1H, Ar-H), 8.11 (s, 1H, Ar-H)
QI-16	5.26 (s, 2H, CH₂), 6.70 (s, 1H, CH), 7.76 (d, J=8.0 Hz, 4H, Ar-H), 8.09 (s, 1H, CH), 8.21 (d, J=8.0 Hz, 2H, Ar-H), 8.29 (d, J=8.0 Hz, 2H, Ar-H)
QII-02	1.04～1.07 (m, 2H, CH₂), 1.19～1.21 (m, 2H, CH₂), 1.84～1.89 (m, 1H, CH), 4.33 (s, 2H, CH₂), 6.53 (s, 1H, CH), 7.91 (s, 1H, CH)
QII-03	1.03 (t, J=7.5 Hz, 3H, CH₃), 1.75～1.79 (m, 2H, CH₂), 2.58 (t, J=7.5 Hz, 2H, CH₂), 4.33 (s, 2H, CH₂), 6.54 (s, 1H, CH), 7.89 (s, 1H, CH)
QII-04	0.94 (t, J=7.5 Hz, 3H, CH₃), 1.40～1.45 (m, 2H, CH₂), 1.69～1.73 (m, 2H, CH₂), 2.59 (t, J=7.5 Hz, 2H, CH₂), 4.33 (s, 2H, CH₂), 6.53 (s, 1H, CH), 7.89 (s, 1H, CH)
QII-05	1.36 (s, 9H, 3×CH₃), 4.33 (s, 2H, CH₂), 6.52 (s, 1H, CH), 7.85 (s, 1H, CH)
QII-06	0.99 (d, J=6.5 Hz, 6H, 2×CH₃), 2.02～2.10 (m, 1H, CH), 4.06 (d, J=6.5 Hz, 2H, CH₂), 4.35 (s, 2H, CH₂), 6.57 (s, 1H, CH), 7.89 (s, 1H, CH)
QII-07	0.88 (t, J=7.5 Hz, 3H, CH₃), 1.28～1.40 (m, 8H, 4×CH₂), 1.71～1.75 (m, 2H, CH₂), 2.59 (t, J=7.5 Hz, 2H, CH₂), 4.33 (s, 2H, CH₂), 6.54 (s, 1H, CH), 7.89 (s, 1H, CH)
QII-08	2.42 (s, 3H, CH₃), 4.37 (s, 2H, CH₂), 6.60 (s, 1H, CH), 7.36～7.39 (m, 1H, Ar-H), 7.44～7.45 (m, 1H, Ar-H), 7.96～7.98 (m, 2H, Ar-H), 8.04 (s, 1H, CH)
QII-09	0.99 (t, J=7.5 Hz, 3H, CH₃), 1.88～1.93 (m, 1H, CH), 2.21～2.27 (m, 1H, CH), 3.75 (t, J=7.5 Hz, H, CH), 4.31 (s, 2H, CH₂), 6.52 (s, 1H, CH), 7.29～7.36 (m, 5H, Ar-H), 7.78 (s, 1H, CH)
QII-10	4.38 (s, 2H, CH₂), 6.62 (s, 1H, CH), 7.77 (d, J=7.5 Hz, 2H, Ar-H), 8.07 (s, 1H, CH), 8.28 (d, J=7.5 Hz, 2H, Ar-H)

Table 2(Continued) 目标化合物的 ¹H NMR数据(CDCl₃, 500 MHz)

Table 2(Continued) ¹H NMR data of target compounds (CDCl₃, 500 MHz)

2.2 化合物抑菌活性

结果(表3)表明，所有供试化合物在50 μg/mL下对苹果炭疽病菌和番茄灰霉病菌的菌丝生长均有不同程度的抑制作用，但总体而言抑菌活性不强。曲酸衍生物中2-苯基丁酸和2-甲基苯甲酸双羟基酰化衍生物(QI-07和QI-11)对苹果炭疽病菌的活性最高，其抑制率分别为79.3%和86.2%；氯曲酸衍生物中QII-04、QII-08和QII-09对番茄灰霉病菌活性最高，其抑制率分别为70.0%、70.0%和76.7%；其余供试化合物的抑制率均在70%以下。

表3 目标化合物对苹果炭疽病菌和番茄灰霉病菌的抑菌活性

Table 3 Antifungal activity of target compounds against G.cingulata and B.cinerea

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农药学学报 2014, Vol.16 Issue (1): 92-96	PDF