WO2022052047A1 - 杀菌剂及杀菌方法 - Google Patents

杀菌剂及杀菌方法 Download PDF

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WO2022052047A1
WO2022052047A1 PCT/CN2020/114821 CN2020114821W WO2022052047A1 WO 2022052047 A1 WO2022052047 A1 WO 2022052047A1 CN 2020114821 W CN2020114821 W CN 2020114821W WO 2022052047 A1 WO2022052047 A1 WO 2022052047A1
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compound
chemical formula
bactericide
curcumin
different
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French (fr)
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胡安仁
施增廉
洪崧壬
陈灏平
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慈济学校财团法人慈济大学
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Priority to CN202080102727.4A priority Critical patent/CN115803310B/zh
Priority to PCT/CN2020/114821 priority patent/WO2022052047A1/zh
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/34—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide
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    • A61K31/33—Heterocyclic compounds
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • C07C49/24—Unsaturated compounds containing keto groups bound to acyclic carbon atoms containing hydroxy groups
    • C07C49/245—Unsaturated compounds containing keto groups bound to acyclic carbon atoms containing hydroxy groups containing six-membered aromatic rings
    • C07C49/248—Unsaturated compounds containing keto groups bound to acyclic carbon atoms containing hydroxy groups containing six-membered aromatic rings having unsaturation outside the aromatic rings
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    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
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    • C07C49/20—Unsaturated compounds containing keto groups bound to acyclic carbon atoms
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
    • C07D307/02—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings
    • C07D307/34—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D307/38—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D307/40—Radicals substituted by oxygen atoms
    • C07D307/46—Doubly bound oxygen atoms, or two oxygen atoms singly bound to the same carbon atom
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
    • C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
    • C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
    • C07D333/06—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to the ring carbon atoms
    • C07D333/22—Radicals substituted by doubly bound hetero atoms, or by two hetero atoms other than halogen singly bound to the same carbon atom

Definitions

  • the present invention relates to a bactericide and a bactericidal method.
  • Curcumin is a yellow pigment extracted from turmeric, and it is the most important curcumin in turmeric, accounting for about 70% of curcumin.
  • curcuminoids are one of the components of the turmeric plant, mainly including demethoxycurcumin (about 10-20%), demethoxycurcumin (about 10%) and Hexahydrocurcumin, etc.
  • Curcumin is used in medical and health food because of its good effect on the human body. Many studies have pointed out that curcumin has various clinical medical effects such as anti-inflammatory, antibacterial, anti-tumor, anti-oxidation, anti-obesity and prevention of chronic diseases. Therefore, in recent years, many curcumin health foods have been sold in the market. In addition, curcumin can also be used as a good photosensitizer. Its absorption wavelength range is 210-510nm, covering the blue part of ultraviolet light and visible light. After irradiation with a specific wavelength, it will generate free radicals, which can cause drug-resistant bacteria to lose activity and resist tumors. the role of cells.
  • curcumin enhances epithelial regeneration and increases fibroblast growth and vascular density on wounds, increases the speed of wound healing and inhibits melanin production in human melanocytes by activating Akt/GSK3 ⁇ , ERK or p38MAPK signaling pathways .
  • the acceptable daily intake (ADI) of curcumin is 0-3mg/kg body weight.
  • the object of the present invention is to further provide a compound that can improve the bactericidal effect more effectively than curcumin, and effectively apply it to a bactericide.
  • the present invention provides a kind of bactericide, it comprises the compound shown in chemical formula I:
  • Ar 1 and Ar 2 are each independently substituted or unsubstituted phenyl, naphthyl, thienyl, furyl or pyridyl, and Ar 1 and Ar 2 are the same or different;
  • the substituent is selected from the group consisting of halogen, hydroxy, alkyl and alkoxy;
  • Ar 1 and Ar 2 are each independently a phenyl group substituted with two substituents, Ar 1 and Ar 2 are different.
  • the compound represented by chemical formula I is selected from at least one of the following compounds 1 to 11:
  • the species that can be killed and/or inhibited by the biocide comprises aerobic bacteria.
  • the species that can be killed and/or inhibited by the biocide comprises Gram-positive bacteria.
  • the gram-positive bacteria comprise Staphylococcus epidermidis or Staphylococcus aureus.
  • the concentration of the compound represented by chemical formula I is 0.5 ppm-1 ppm.
  • the compounds used for sterilization of the present invention are all artificially synthesized, have high purity, and have better sterilization effects compared to three kinds of mixtures in natural curcumin.
  • curcumin and its derivatives are known to have bactericidal effects, they are not necessarily effective against any strain.
  • the curcuminoid used in the present invention has a relatively high bactericidal ability for aerobic bacterial species.
  • Figure 1 shows the bactericidal effect of curcumin or curcuminoids on Staphylococcus epidermidis.
  • Figure 2 shows the SEM images of the effect of compound 8 on the structure of Staphylococcus epidermidis before and after irradiation with blue light, wherein (a) is before irradiation, (b) is after 1 minute of irradiation and (c) is after 5 minutes of irradiation.
  • the term "sterilization” means having the ability to kill, destroy or destroy bacterial species, reduce the number of bacterial species and/or inhibit the proliferation of bacterial species.
  • the present invention provides a kind of bactericide, it comprises the compound shown in chemical formula I:
  • Ar 1 and Ar 2 are each independently substituted or unsubstituted phenyl, naphthyl, thienyl, furyl or pyridyl, and Ar 1 and Ar 2 are the same or different;
  • the substituent is selected from the group consisting of halogen, hydroxy, alkyl and alkoxy;
  • Ar 1 and Ar 2 are each independently a phenyl group substituted with two substituents, Ar 1 and Ar 2 are different.
  • the LED tube was set up on a blue light device made of polystyrene, and kept a distance of about 5 mm from the test tube to be irradiated.
  • the above 17 kinds of synthetic curcuminoids were weighed to 1 mg, dissolved in 0.5 ml of DMSO solvent, and used as a stock solution standard of 100% DMSO 2000 ppm. Take 10 ⁇ l of the stock solution and add it to 990 ⁇ l of LB medium (diluted 100 times), then take 100 ⁇ l of curcuminoid diluted 100 times and add it to 900 ⁇ l of LB medium (diluted 1000 times), and finally take the 1000 times diluted curcuminoid. 1 ml of curcumin was added to 1 ml of LB medium as the reagent to be reacted (2 ml, finally diluted 2000 times in total, ie 1 ppm), which was used as the experimental group.
  • DMSO supplemented with curcumin but also diluted 2000 times with LB medium was used as a comparison group
  • DMSO without addition of curcumin or curcuminoids but also diluted 2000 times with LB medium was used as a control group (control).
  • the final diluted curcuminoid 2ml take 1ml and add two tubes to the centrifuged thalline that has removed the supernatant, place in the test tube of 25ml capacity after evenly mixing, and put one tube into the blue light device and irradiate for 1 minute ( 1min), and then carry out the drip-coating method. Another tube, unirradiated (no light) was used as a control.
  • the comparison group ie, curcumin added
  • the control group ie, no curcumin or curcuminoid added
  • Step 4 Calculation of sterilization rate of blue light and photosensitizer
  • T is the number of colonies in the experimental group or the comparison group that are not irradiated (no light) or irradiated for 1 minute (1 min), or the control group is irradiated for 1 minute (1 min), and C is the control group (that is, without adding curcumin or curcuminoid) ) unirradiated (no light) colonies.
  • Part 1 Staphylococcus epidermidis and Staphylococcus aureus
  • FIG. 1 shows the sterilization of S. epidermidis by the 17 synthetic curcuminoids described above in the absence/presence of photodynamic action (ie, no blue light irradiation (no light) or blue light irradiation for 1 minute (1 min)).
  • the sterilization rate was calculated based on the number of colonies when no curcumin or curcuminoid was added (control group) and without blue light irradiation (no light), so the value was zero.
  • the inventors further repeated three times for the effects on Staphylococcus epidermidis and Staphylococcus aureus for the five types of curcumin with the best bactericidal effect (respectively compound 1, compound 2, compound 5, compound 8 and compound 9).
  • the results are shown in Table 1 and Table 2, respectively. It can be found that Compound 1, Compound 8 and Compound 9 have the best bactericidal effects on Staphylococcus epidermidis and Staphylococcus aureus. Among them, compound 8 and compound 9 both have excellent bactericidal effect on Staphylococcus epidermidis or Staphylococcus aureus, and the sterilization rate can reach more than 99%.
  • Figure 2 shows the SEM images of the effect of compound 8 on the structure of Staphylococcus epidermidis before (a) irradiation, (b) 1 minute after irradiation, and (c) 5 minutes after irradiation with blue light, respectively. It can be clearly seen that compound 8 has the killing ability to cause perforation to Staphylococcus epidermidis after irradiation.
  • Compound 1, Compound 8, and Compound 9 were prepared as 2000 ppm stock solution standards in DMSO, respectively, and stored in an unirradiated room temperature (25°C) environment. During the stability test, these stock solutions were diluted 100 times with DMSO (ie, the concentration was 20 ppm), and after standing for 48 hours, the color of compound 1 could be observed slightly changed, while the color of compound 8 and compound 9 changed slightly. without. Take 2 ⁇ l respectively and measure the absorbance of each compound at 220-750 nm on an ultra-micro spectrophotometer.
  • the curcuminoid compound 8 which has excellent bactericidal effect against Staphylococcus epidermidis or Staphylococcus aureus, further explored its effect on Escherichia coli.
  • Method 1 Dilute the stock solution standard by 2000 times and the bacterial number concentration is about 10 9 CFU/ml

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Abstract

本发明提供一种杀菌剂及杀菌方法，其中杀菌剂包含化学式（I）所示的化合物，其中，Ar1及Ar2各自独立地为取代基取代或未取代的苯基、萘基、噻吩基、呋喃基或吡啶基，且Ar1及Ar2相同或不同；该取代基选自由卤素、羟基、烷基及烷氧基所组成的组；当Ar1及Ar2各自独立地为两个取代基取代的苯基时，Ar1及Ar2不同。

Description

杀菌剂及杀菌方法 技术领域
本发明是关于杀菌剂及杀菌方法。
背景技术
姜黄素(curcumin)是一种从姜黄(turmeric)中提取得到的黄色色素，为姜黄中最主要的姜黄色素，约占姜黄色素的70％。除了姜黄素之外，类姜黄素(curcuminoids)为姜黄植物中的成分之一，主要包括去甲氧基姜黄素(约10-20％)、去二甲氧基姜黄素(约10％)和六氢姜黄素等。
Figure PCTCN2020114821-appb-000001
姜黄素因对人体具有良好的作用而被应用在医疗及保健食品上。有许多的研究指出姜黄素具有抗发炎、抗菌、抗肿瘤、抗氧化、抗肥胖以及预防慢性疾病等各种临床医疗功效，因此近年来已有许多姜黄素的保健食品于市面上贩卖。另外姜黄素也可作为良好的光敏剂，其吸收波长范围为210-510nm，涵盖了紫外光与可见光的蓝光部分，经由特定波长照射后会产生自由基，可造成抗药性细菌失去活性和抗肿瘤细胞的作用。除此之外，姜黄素在伤口上可增强上皮再生以及增加成纤维细胞生长与血管密度，增加伤口愈合的速度以及通过活化Akt/GSK3β、ERK或p38MAPK信号传导途径抑制人体黑色素细胞中的黑色素生成。根据JECFA(联合国和世界卫生组织联合食品添加剂专家委员会)和EFSA(欧洲食品安全局)的报告，姜黄素的每日允许摄入量(ADI)为0-3mg/kg体重。
发明内容
本发明的目的在于进一步提供一种相较于姜黄素能更有效提升杀菌效果的化合物，并将其有效应用于杀菌剂。
本发明提供一种杀菌剂，其包含化学式I所示的化合物：
Figure PCTCN2020114821-appb-000002
其中，
Ar 1及Ar 2各自独立地为取代基取代或未取代的苯基、萘基、噻吩基、呋喃基或吡啶基，且Ar 1及Ar 2相同或不同；
该取代基选自由卤素、羟基、烷基及烷氧基所组成的组；
当Ar 1及Ar 2各自独立地为两个取代基取代的苯基时，Ar 1及Ar 2不同。
优选地，化学式I所示的化合物选自下列化合物1至化合物11中的至少之一：
Figure PCTCN2020114821-appb-000003
Figure PCTCN2020114821-appb-000004
优选地，杀菌剂可杀灭和/或抑制的菌种包含需氧菌。
优选地，杀菌剂可杀灭和/或抑制的菌种包含革兰氏阳性菌。
优选地，革兰氏阳性菌包含表皮葡萄球菌或金黄色葡萄球菌。
优选地，化学式I所示的化合物的浓度为0.5ppm-1ppm。
本发明还提供一种杀菌方法，其包括：将上述化学式I所示的化合物与菌种接触；以及对该化合物施予波长为410-510nm(λ max＝462nm)的光源照射。
本发明的杀菌剂及杀菌方法至少具有下述优点之一：
1、本发明用于杀菌的化合物全部是人工合成，纯度高，相较于天然的姜黄素内的三种混合物，具有更佳的杀菌效果。
2、虽然已知姜黄素及其衍生物具有杀菌效果，但并非对任何菌种均必然有效。本发明所使用的类姜黄素对于需氧性的菌种有相当高的杀菌能力。
附图说明
此处所说明的附图用来提供对本申请的进一步理解，其构成本 申请的一部分，但并不构成对本申请的不当限制。附图中：
图1为姜黄素或类姜黄素对表皮葡萄球菌的杀菌效果。
图2为化合物8经蓝光照射前和照射后，对表皮葡萄球菌结构影响的SEM图，其中(a)为照射前、(b)为照射1分钟后及(c)为照射5分钟后。
图3为类姜黄素的稳定性测试结果图，其中(a)为化合物1、(b)为化合物8及(c)为化合物9。
具体实施方式
为使本申请的目的、技术方案和优点更加清楚，下面将参照相关附图并结合实施例对本发明的技术方案进行清楚、完整地描述，其中所描述的实施例仅是本申请一部分，而不是全部的实施例。
除非特别定义，否则所有本文中所用的技术及科学名词均与本领域技术人员所通常理解的意义相同。如果本文中所用定义与其他公开文献中所记载定义矛盾或不一致，则应以本文所用的定义为准。
如本文中所用的，术语“含”、“包含”、“含有”、“具有”或其任何其他变型，意欲涵盖非排他性的包括。
数值或参数给出了范围、优选范围或上限值与下限值之列表时，应将其理解为具体地公开由任何范围上限或优选值与任何范围下限或优选值的任何配对所形成的所有范围，无论这些范围是否分别公开。例如，当叙述一个范围为“1至5”时，所叙述的范围应被理解为包含范围“1至4”、“1至3”、“1至2”、“1至2与4至5”、“1至3与5”等等。若本说明书中叙述一个数值范围，则除非另有说明，该范围意欲包含其端点以及该范围内的所有整数和小数。
此外，除非另有明确说明，“或”意指包含性的“或”而非指排除性的“或”。例如，条件A“或”B满足于下述情况中任何一种：A成立(或存在)且B不成立(或不存在)、A不成立(或不存在)且B成立(或存在)、以及A和B两者皆成立(或存在)。
同样地，除非上下文明确说明，否则位于本文的元素或成份之 前的不定冠词“一”、“一个”及“一种”旨在非限制性地说明所述元素或成份的示例数目(即出现数)。因此，“一”、“一个”及“一种”应理解为包含一个或至少一个，且所述元素或成分的单数词形也包含复数形式。
本文中所使用的“杀菌”一词，意指具有杀灭、消灭或破坏菌种、降低菌种数量和/或抑制菌种增生的能力。
在一个实施方案中，本发明提供一种杀菌剂，其包含化学式I所示的化合物：
Figure PCTCN2020114821-appb-000005
其中，
Ar 1及Ar 2各自独立地为取代基取代或未取代的苯基、萘基、噻吩基、呋喃基或吡啶基，且Ar 1及Ar 2相同或不同；
该取代基选自由卤素、羟基、烷基及烷氧基所组成的组；
当Ar 1及Ar 2各自独立地为两个取代基取代的苯基时，Ar 1及Ar 2不同。
实施例
实施例中，使用了17种合成的类姜黄素与作为比较例的姜黄素(Curcumin I)进行实验比较，其结构分别如下所示：
Figure PCTCN2020114821-appb-000006
Figure PCTCN2020114821-appb-000007
实验方法
菌株培养
菌种1：由-80℃冰箱中取出表皮葡萄球菌(Staphylococcus epidermidis TCU-1 BCRC 81267)，以接种环蘸取菌液，于LB培养基上划三区，置于37℃的培养箱中12小时，次日以接种环取出盘上的一个菌落于10ml的LB培养基中在37℃培养箱中培养12小时，测得吸光度OD 600nm＝1后，细菌数量浓度约为10 9CFU/ml，以2000xg进行离心10分钟，去除上清液。
菌种2：由-80℃冰箱中取出金黄色葡萄球菌(Staphylococcus aureus subsp.aureus TCU-2 BCRC81268)，以接种环蘸取菌液，于LB培养基上划三区，置于37℃培养箱中12小时，次日以接种环取出盘上的一个菌落于10ml的LB培养基中在37℃培养箱中培养12小时，测得吸光度OD 600nm＝1后，细菌数量浓度约为10 9CFU/ml，以2000xg进行离心10分钟，去除上清液。
菌种3：由-80℃冰箱中取出大肠杆菌(Escherichia coli)，以接种环蘸取菌液，于LB培养基上划三区，置于37℃培养箱中12小时，次日以接种环取出盘上的一个菌落于10ml的LB培养基中在37℃培养箱中培养12小时，测得吸光度OD 600nm＝1后，细菌数量浓度约为10 9CFU/ml。此时再连续稀释1000倍，吸取菌液200μl至新的离心管中，再加入800μl的LB培养基，此时细菌数量约为5 x 10 5CFU/ml，进行离心(2000xg，10分钟)。
蓝光装置与照射方式
LED T8灯管：光波长为410-510nm(λ max＝462nm)，功率最大值为5W。
将LED灯管架设于聚苯乙烯制蓝光装置上，并与待照射的试管保持距离约5mm。
实验流程
步骤1、实验组与对照组的样品配制
将上述17种合成的类姜黄素称重1mg，溶解于0.5ml的 DMSO溶剂中，作为100％DMSO 2000ppm的原液标准品。取出10μl的原液加入990μl的LB培养基中(稀释100倍)，再从稀释100倍的类姜黄素取出100μl加入于900μl的LB培养基中(稀释1000倍)，最终再取稀释1000倍的类姜黄素1ml加入1ml的LB培养基中，作为待反应试剂(2ml，最终共稀释2000倍，即1ppm)，其作为实验组。此外，以添加姜黄素但同样以LB培养基稀释2000倍的DMSO作为比较组，且以未添加姜黄素或类姜黄素但同样以LB培养基稀释2000倍的DMSO作为对照组(control)。
步骤2、蓝光诱导光敏剂杀菌
将最终稀释的类姜黄素2ml，分别取1ml加入两管已去除上清液的经离心菌体中，均匀混合后置于25毫升容量的试管中，一管放入蓝光装置中照射1分钟(1min)，再进行点滴涂板法。另一管，以未照射(无光照)作为对照。比较组(即，添加姜黄素)及对照组(即，未添加姜黄素或类姜黄素)也配制有照射1分钟或未照射的两个试管。
步骤3、点滴涂板法
将上述反应完的样品以LB培养基作为溶剂，系列稀释至10E-7，于10E-4、10E-5、10E-6、10E-7分别取10μl滴于LB培养基中，于37℃下培养12小时后，以10E-5作为细菌的计数并拍照记录结果。
步骤4、蓝光及光敏剂的杀菌率计算
杀菌率＝{1-T (CFU/ml)/C (CFU/ml)}×100％
其中，T为实验组或比较组未照射(无光照)或照射1分钟(1min)，或对照照射1分钟(1min)的菌落数，C为对照组(即，未添加姜黄素或类姜黄素)未照射(无光照)的菌落数。
实验结果
第一部分：表皮葡萄球菌与金黄色葡萄球菌
请参见图1，其显示上述17种合成的类姜黄素在无/有光动力作用(即，无蓝光照射(无光照)或蓝光照射1分钟(1min))的情况下对表皮葡萄球菌的杀菌率，其中，杀菌率以未添加姜黄素或类 姜黄素(对照组)且未经蓝光照射(无光照)时的菌落数作为计算基础，因此数值为零。从图1中可发现，化合物1、化合物2、化合物5、化合物8和化合物9经照射1分钟(样品名称分别为1(1min)、2(1min)、5(1min)、8(1min)和9(1min))，相较于其他化合物具有明显较好的杀菌效果。
发明人进一步针对杀菌效果最佳的这五种类姜黄素(分别为化合物1、化合物2、化合物5、化合物8和化合物9)，就其对表皮葡萄球菌和金黄色葡萄球菌的影响再进行三次重复实验，其结果分别如表1和表2所示，可发现化合物1、化合物8及化合物9对于表皮葡萄球菌及金黄色葡萄球菌的杀菌效果均最佳。其中，化合物8和化合物9不管对于表皮葡萄球菌或金黄色葡萄球菌均具有极佳的杀菌效果，杀菌率均可达99％以上。图2分别为化合物8在蓝光(a)照射前、(b)照射后1分钟和(c)照射后5分钟，对表皮葡萄球菌结构影响的SEM图。可明显看出，化合物8经照射后，对表皮葡萄球菌具有造成穿孔的杀伤力。
表1、五种合成的类姜黄素对于表皮葡萄球菌的影响
Figure PCTCN2020114821-appb-000008
Figure PCTCN2020114821-appb-000009
表2、五种合成的类姜黄素对于金黄色葡萄球菌的影响
Figure PCTCN2020114821-appb-000010
Figure PCTCN2020114821-appb-000011
进一步地，遵循上述的实验方法进行杀菌测试，不同仅在于将化合物1、化合物8及化合物9的浓度下降至0.5ppm，所得的实验结果如表3所示。可发现化合物8和化合物9即使降低浓度，仍有很好的杀菌效果。
表3、浓度0.5ppm的化合物1和化合物8和化合物9对于表皮葡萄球菌葡的影响
Figure PCTCN2020114821-appb-000012
类姜黄素稳定性测试
将化合物1、化合物8及化合物9分别以DMSO配制为2000ppm的原液标准品，并将其存放于未照射的室温(25℃)环境中。稳定性实验测试时，将这些原液标准品再以DMSO稀释100倍(即，浓度为20ppm)，并放置48小时后，肉眼可以观察到化合物1的颜色略有改变，而化合物8与化合物9则无。分别取2μl于超微量分光光度计上测量各化合物于220-750nm下的吸光度。如图3的结果显示，化合物8(最大吸收波长440nm)与化合物9(最大吸收波长426nm)在最大波长下的吸光度下降约0.8％与1.3％(分别如图(b)和(c)所示)，而化合物1(最大吸收波长426nm)在最大波长下的吸光度则下降了约6.4％。因此，根据此结果，可发现化合物8和化合物9相较于化合物1具有更好的稳定性。
第二部分：大肠杆菌
对于表皮葡萄球菌或金黄色葡萄球菌均具有极佳杀菌效果的类姜黄素化合物8，进一步探讨其对于大肠杆菌的影响。
方法一：将原液标准品稀释2000倍且细菌数量浓度约为10 9CFU/ml
如上述步骤1所记载的，将化合物8称重1mg，溶解于0.5ml的DMSO溶剂中，作为100％DMSO 2000ppm的原液标准品，再通过LB培养基稀释2000倍作为待反应试剂(1ppm)。此外，大肠杆菌的培养则如上述菌种培养的菌种3所记载，测得吸光度OD 600nm＝1后，以2000xg进行离心10分钟，去除上清液，取得细菌数量浓度约10 9CFU/ml。对照组除未添加化合物8外，其他条件均与实验组相同，且后续实验流程同上述步骤2至4。其实验结果如表4所示，在此条件下，化合物8对于大肠杆菌的杀菌效果有限。
表4、类姜黄素化合物8(1ppm)对于大肠杆菌的光动力杀菌效果
Figure PCTCN2020114821-appb-000013
方法二：将原液标准品稀释1000倍且细菌数量浓度约为10 9CFU/ml
如上述步骤1所记载，将化合物8称重1mg，溶解于0.5ml的DMSO溶剂中，作为100％DMSO 200ppm的原液标准品，再通过LB培养基稀释1000倍作为待反应试剂。此外，大肠杆菌的培养则如上述菌种培养的菌种3所记载，测得吸光度OD 600nm＝1后，以2000xg进行离心10分钟，去除上清液，取得细菌数量浓度约为10 9CFU/ml。对照组除未添加化合物8外，其他条件均与实验组相同，且后续实验流程同上述步骤2至4。相较于方法一，方法二进一步提高类姜黄素化合物8的浓度至2ppm，其实验结果如表5所示。在此条件下，化合物8对于大肠杆菌的杀菌效果稍加提升。
表5、类姜黄素化合物8(2ppm)对于大肠杆菌的光动力杀菌效果
Figure PCTCN2020114821-appb-000014
以上所述仅为例示性的，而非为限制性的。任何未脱离本发明的精神与范畴，而对其进行的等效修改或变更，均应包含于后附的权利要求的范围中。

Claims (12)

  1. 一种杀菌剂，其包含化学式I所示的化合物：
    Figure PCTCN2020114821-appb-100001
    其中，
    Ar 1及Ar 2各自独立地为取代基取代或未取代的苯基、萘基、噻吩基、呋喃基或吡啶基，且Ar 1及Ar 2相同或不同；
    该取代基选自由卤素、羟基、烷基及烷氧基所组成的组；
    当Ar 1及Ar 2各自独立地为两个取代基取代的苯基时，Ar 1及Ar 2不同。
  2. 如权利要求1所述的杀菌剂，其中该化学式I所示的化合物选自下列化合物1至化合物11中的至少之一：
    Figure PCTCN2020114821-appb-100002
    Figure PCTCN2020114821-appb-100003
  3. 如权利要求1所述的杀菌剂，其可抑制和/或杀灭的菌种包含需氧菌。
  4. 如权利要求1所述的杀菌剂，其可抑制和/或杀灭的菌种包含革兰氏阳性菌。
  5. 如权利要求4项所述的杀菌剂，其中该革兰氏阳性菌包含表皮葡萄球菌或金黄色葡萄球菌。
  6. 如权利要求1所述的杀菌剂，其中该化学式I所示的化合物的浓度为0.5ppm-1ppm。
  7. 一种杀菌方法，其包括：
    将化学式I所示的化合物与菌种接触；以及
    对该化合物施予蓝光范围波长的光源照射；
    Figure PCTCN2020114821-appb-100004
    其中，
    Ar 1及Ar 2各自独立地为取代基取代或未取代的苯基、萘基、噻吩基、呋喃基或吡啶基，且Ar 1及Ar 2相同或不同；
    该取代基选自由卤素、羟基、烷基及烷氧基所组成的组；
    当Ar 1及Ar 2各自独立地为两个取代基取代的苯基时，Ar 1及Ar 2不同。
  8. 如权利要求7所述的方法，其中该化学式I所示的化合物选自下列化合物1至化合物11中的至少之一：
    Figure PCTCN2020114821-appb-100005
  9. 如权利要求7所述的方法，其中该菌种包含需氧菌。
  10. 如权利要求7所述的方法，其中该菌种包含革兰氏阳性菌。
  11. 如权利要求10所述的方法，其中该革兰氏阳性菌包含表皮葡萄球菌或金黄色葡萄球菌。
  12. 如权利要求7所述的方法，其中该化学式I所示的化合物的浓度为0.5ppm-1ppm。
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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108347927A (zh) * 2015-08-26 2018-07-31 雷根斯堡大学临床中心 1，7-二芳基-1，6-庚二烯-3，5-二酮衍生物、其制备方法和用途

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108347927A (zh) * 2015-08-26 2018-07-31 雷根斯堡大学临床中心 1，7-二芳基-1，6-庚二烯-3，5-二酮衍生物、其制备方法和用途

Non-Patent Citations (10)

* Cited by examiner, † Cited by third party
Title
CHEN, LIMIN ET AL.: "Synthesis and in vitro Antioxidant Activity of Curcumin Analogs", NATURAL PRODUCT RESEARCH AND DEVELOPMENT, vol. 23, no. 4, 15 August 2011 (2011-08-15), pages 722 - 725, XP055911242, ISSN: 1001-6880 *
DING, TAO ET AL.: "Effect of Curcumin-mediated Photodynamic Therapy on Methicillin-resistant Staphylococcus Aureus and Biofilm in Vitro", CHINESE JOURNAL OF EXPERIMENTAL SURGERY, vol. 36, no. 6, 8 June 2019 (2019-06-08), pages 1 - 46, XP055911245, ISSN: 1001-9030 *
HAUKVIK T, BRUZELL E, KRISTENSEN S, TØNNESEN H H: "A screening of curcumin derivatives for antibacterial phototoxic effects Studies on curcumin and curcuminoids. XLIII", PHARMAZIE, vol. 66, no. 1, 15 January 2011 (2011-01-15), DE , pages 69 - 74, XP055911205, ISSN: 0031-7144, DOI: 10.1691/ph.2011.0154 *
LUO JIAOYANG, YANG MEIHUA: "Demethoxycurcumin: a potential antimicrobial agent : Exposure by microcalorimetry and modified broth microdilution method", JOURNAL OF THERMAL ANALYSIS AND CALORIMETRY, vol. 115, no. 3, 1 March 2014 (2014-03-01), NL , pages 2331 - 2338, XP055911215, ISSN: 1388-6150, DOI: 10.1007/s10973-013-3103-6 *
M. AKRAM KHAN; RIYAD EL-KHATIB; K.D. RAINSFORD; M.W. WHITEHOUSE;: "Synthesis and anti-inflammatory properties of some aromatic and heterocyclic aromatic curcuminoids", BIOORGANIC CHEMISTRY, vol. 40, US , pages 30 - 38, XP028443386, ISSN: 0045-2068, DOI: 10.1016/j.bioorg.2011.11.004 *
POLAQUINI C. R. ET AL.: "Antibacterial Activity of 3, 3'-Dihydroxycurcumin (DHC) is Associated with Membrane Perturbation", BIOORGANIC CHEMISTRY, vol. 90, 7 June 2019 (2019-06-07), XP085760154, ISSN: 0045-2068, DOI: 10.1016/j.bioorg.2019.103031 *
SINDHU, S.; THOMACHAN, SEENA; JOHN V. D.: "Synthesis, Characterization, and Biological Studies of 1,7-Diheteroaryl-1,6-Heptadiene-3,5-Dione and Their Metal Complexes", HETEROCYCLIC LETTERS, vol. 5, no. 2, 31 May 2015 (2015-05-31), India, pages 285 - 293, XP009535229, ISSN: 2230-9632 *
SOARES J. M. ET AL.: "Optimization for Microbial Incorporation and Efficiency of Photodynamic Therapy Using Variation on Curcumin Formulation", PHOTODIAGNOSIS AND PHOTODYNAMIC THERAPY, vol. 29, 7 January 2020 (2020-01-07), XP086099340, ISSN: 1572-1000, DOI: 10.1016/j.pdpdt.2020.101652 *
YANG MING-YEH; CHANG KAI-CHIH; CHEN LIANG-Yü; HU ANREN: "Low-dose blue light irradiation enhances the antimicrobial activities of curcumin againstPropionibacterium acnes", JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B: BIOLOGY, vol. 189, 26 September 2018 (2018-09-26), CH , pages 21 - 28, XP085552953, ISSN: 1011-1344, DOI: 10.1016/j.jphotobiol.2018.09.021 *
ZHENG DANTONG, HUANG CHONGXING, HUANG HAOHE, ZHAO YUAN, KHAN MUHAMMAD RAFI ULLAH, ZHAO HUI, HUANG LIJIE: "Antibacterial Mechanism of Curcumin: A Review", CHEMISTRY & BIODIVERSITY, HELVETICA CHIMICA ACTA, vol. 17, no. 8, 1 August 2020 (2020-08-01), CH , XP055911251, ISSN: 1612-1872, DOI: 10.1002/cbdv.202000171 *

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