JP2022102189A - Antibacterial and antiviral agent - Google Patents
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- JP2022102189A JP2022102189A JP2020216778A JP2020216778A JP2022102189A JP 2022102189 A JP2022102189 A JP 2022102189A JP 2020216778 A JP2020216778 A JP 2020216778A JP 2020216778 A JP2020216778 A JP 2020216778A JP 2022102189 A JP2022102189 A JP 2022102189A
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- zinc oxide
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- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 57
- 239000003242 anti bacterial agent Substances 0.000 title claims abstract description 31
- 239000003443 antiviral agent Substances 0.000 title claims abstract description 29
- XLOMVQKBTHCTTD-UHFFFAOYSA-N Zinc monoxide Chemical compound [Zn]=O XLOMVQKBTHCTTD-UHFFFAOYSA-N 0.000 claims abstract description 84
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- 239000000284 extract Substances 0.000 claims abstract description 27
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Abstract
Description
本発明は一般に、殺菌やウイルスの不活性化を行う薬剤に関するものであり、詳しくは、長期間にわたって抗菌・抗ウイルスの活性が維持される抗菌・抗ウイルス薬剤に関する。 The present invention generally relates to an agent that sterilizes or inactivates a virus, and more particularly to an antibacterial / antiviral agent that maintains antibacterial / antiviral activity for a long period of time.
２０２０年の新型コロナウイルス感染症（ＣＯＶＩＤ－１９）のパンデミック以降、ウイルスなどによる感染症の予防は、生活の上で必須になってきている。 Since the pandemic of the new coronavirus infection (COVID-19) in 2020, prevention of infectious diseases caused by viruses and the like has become indispensable in daily life.
感染症の予防には、手に触れるものをエタノールなどのアルコールなどによって消毒しておくことが有効である。しかしながらアルコールは、ノロウイルス等のエンベロープを有しないウイルスの不活性化ができない、短時間で揮発し効果が長続きしないなどの問題があった。 To prevent infectious diseases, it is effective to disinfect anything that comes into contact with hands with alcohol such as ethanol. However, alcohol has problems that it cannot inactivate non-enveloped viruses such as norovirus, and that it volatilizes in a short time and its effect does not last long.
また、次亜塩素酸水も広く利用されているが、殺菌に時間がかかる、持続的な殺菌やウイルスの不活性化を行うことができないなどの問題があった。 Hypochlorite water is also widely used, but has problems such as time-consuming sterilization, continuous sterilization, and inability to inactivate the virus.
ところで、本発明者らは、有機酸亜鉛を５０００～１００００ｐｐｍの濃度で含む薬剤が、鳥インフルエンザウイルスを不活性化する能力があることを見出し、ウイルス不活性化薬剤として利用することを提案した（特許文献１）。また、本発明者らは、この有機酸亜鉛を含む薬剤は、ウイルス以外に、各種の細菌・真菌に対しても抗菌力を有することを示している（特許文献２）。 By the way, the present inventors have found that a drug containing zinc organic acid at a concentration of 5000 to 10000 ppm has an ability to inactivate avian influenza virus, and have proposed to use it as a virus inactivating agent (). Patent Document 1). In addition, the present inventors have shown that the drug containing zinc organic acid has antibacterial activity against various bacteria and fungi in addition to viruses (Patent Document 2).
最近では多くの消毒薬剤が上市されているが、安全で且つ効果が長期間持続する消毒薬剤に対する要望はより高まっている。 Recently, many disinfectants have been put on the market, but there is a growing demand for disinfectants that are safe and have long-lasting effects.
本発明は、長時間にわたって抗菌・抗ウイルス作用が持続する安心安全な抗菌・抗ウイルス薬剤を提供することを目的とする。 An object of the present invention is to provide a safe and secure antibacterial / antiviral agent whose antibacterial / antiviral action is sustained for a long period of time.
上記課題を解決するための、本発明に係る抗菌・抗ウイルス薬剤は、酸化亜鉛ナノ粒子と、柑橘類種子抽出物とが、水系の溶媒に分散されてなる抗菌・抗ウイルス薬剤であって、前記柑橘類種子抽出物の濃度が２５００～１００００ｐｐｍである。 The antibacterial / antiviral agent according to the present invention for solving the above problems is an antibacterial / antiviral agent in which zinc oxide nanoparticles and a citrus seed extract are dispersed in an aqueous solvent. The concentration of citrus seed extract is 2500-10000 ppm.
酸化亜鉛粒子は、好ましくは直径が８０ｎｍ以下、より好ましくは直径が４０～７０ｎｍの超微粒子（ナノ粒子）とする。このような超微粒子状の酸化亜鉛は、水系の溶媒に良く分散する。ここで、金属イオンには、Ｈｇ＞Ａｇ＞Ｃｕ＞Ｚｎ＞Ｆｅ＞ＴｉＯ2の順で殺菌力があるといわれている。このため、銀系抗菌剤が用いられることが多いが、酸化亜鉛を超微粒子にすることにより、銀に劣らない抗菌性を示すようになる。超微粒子の酸化亜鉛の抗菌メカニズムは銀イオンと同じと考えられ、金属の毒性、殺菌性によるものでなく、空気中あるいは水中の酸素の一部を活性酸素化し、ウイルスのエンベロープや細菌の細胞膜等を破壊すると考えられる。 The zinc oxide particles are preferably ultrafine particles (nanoparticles) having a diameter of 80 nm or less, and more preferably 40 to 70 nm in diameter. Such ultrafine zinc oxide is well dispersed in an aqueous solvent. Here, it is said that metal ions have bactericidal activity in the order of Hg>Ag>Cu>Zn>Fe> TiO 2 . For this reason, a silver-based antibacterial agent is often used, but by making zinc oxide into ultrafine particles, the antibacterial property is not inferior to that of silver. The antibacterial mechanism of ultrafine zinc oxide is considered to be the same as that of silver ions, and it is not due to the toxicity and bactericidal properties of metals. Is thought to destroy.
酸化亜鉛をナノ粒子として使用することにより、比表面積が大きくなり、ウイルスの表面での接触が拡大されて、ウイルスの増殖が抑制されると考えられる。なお、ウイルスの径は様々であるが、おおむね５０～２００ｎｍである。ウイルスの径に近い、あるいはウイルスの径よりも小さい径を有する超微粒子の酸化亜鉛を選んで、水系の溶媒に分散させることにより、エンベロープの有無にかかわらずウイルスを不活性化することができる。 It is considered that the use of zinc oxide as nanoparticles increases the specific surface area, expands the contact on the surface of the virus, and suppresses the growth of the virus. The diameter of the virus varies, but it is generally 50 to 200 nm. By selecting ultrafine zinc oxide having a diameter close to or smaller than the diameter of the virus and dispersing it in an aqueous solvent, the virus can be inactivated regardless of the presence or absence of an envelope.
また、柑橘類種子抽出物は、抗菌、抗ウイルス効果を有することが知られている。しかしながら、柑橘類種子抽出物の活性は、対象物に噴霧したり拭いたりした後には比較的短期間（およそ５～３０日）でなくなってしまう。 In addition, citrus seed extract is known to have antibacterial and antiviral effects. However, the activity of the citrus seed extract disappears in a relatively short period of time (approximately 5-30 days) after spraying or wiping the object.
また、酸化亜鉛は柑橘類種子抽出物との反応性が低く、その効果を損なうような副生成物が生じることがない。 In addition, zinc oxide has low reactivity with citrus seed extract and does not produce by-products that impair its effectiveness.
さらに、酸化亜鉛は医薬品や日焼け止めなどに含まれる成分であり、また柑橘類種子抽出物は食品添加物として使用されている。つまり、これらの成分は人体に対する安全性に優れる。 Furthermore, zinc oxide is a component contained in pharmaceuticals and sunscreens, and citrus seed extract is used as a food additive. That is, these components are excellent in safety to the human body.
本発明では、酸化亜鉛ナノ粒子と、柑橘類種子抽出物とを組み合わせて用いているが、この構成によると、酸化亜鉛ナノ粒子単独、柑橘類種子抽出物単独よりも高い抗菌、抗ウイルス効果が、一か月以上の長期にわたって得られるようになる。これは、酸化亜鉛ナノ粒子が柑橘類種子抽出物の安定性を高めるように作用することによると考えられる。これにより、比較的低濃度の柑橘類種子抽出物を用いた、アルコールフリーで高い抗菌・抗ウイルス力を持続的に発揮する安全な抗菌・抗ウイルス薬剤を実現できる。 In the present invention, zinc oxide nanoparticles and citrus seed extract are used in combination. According to this configuration, zinc oxide nanoparticles alone and citrus seed extract alone have higher antibacterial and antiviral effects. It will be available for a long period of more than a month. This is believed to be due to the zinc oxide nanoparticles acting to enhance the stability of the citrus seed extract. This makes it possible to realize a safe antibacterial / antiviral agent that is alcohol-free and continuously exerts high antibacterial / antiviral activity using a citrus seed extract having a relatively low concentration.
上記水系の溶媒は、グルコン酸、酢酸、クエン酸、酒石酸、リンゴ酸、乳酸、γ－ヒドロキシ酪酸、フマール酸等を含んだものとすることができ、中でも水に溶解させると一部ラクトン化する有機酸（γ－ヒドロキシ酪酸、グルコン酸など）を水に溶かしたものであることが好ましい。これらの有機酸は人体にとって有害ではないとともに、柑橘類種子抽出物の効果を高めるように作用する。 The aqueous solvent may contain gluconic acid, acetic acid, citric acid, tartaric acid, malic acid, lactic acid, γ-hydroxybutyric acid, fumaric acid, etc. It is preferably a solution of an organic acid (γ-hydroxybutyric acid, gluconic acid, etc.) in water. These organic acids are not harmful to the human body and act to enhance the effectiveness of the citrus seed extract.
ここで、酸化亜鉛ナノ粒子は有機酸水溶液中ではその一部が有機酸と反応して、有機酸亜鉛となり、水に溶解するようになる。これにより、次のような効果が得られる。 Here, a part of the zinc oxide nanoparticles reacts with the organic acid in the organic acid aqueous solution to become zinc organic acid, which becomes soluble in water. As a result, the following effects can be obtained.
ここでは、有機酸としてグルコン酸を用いた場合を例として説明する。酸化亜鉛はグルコン酸水溶液中ではその一部がグルコン酸と反応して、グルコン酸亜鉛（グルコン酸イオンと亜鉛イオン）となり、水に溶解するようになる。亜鉛イオンは銀イオンよりも殺菌力が弱いとされているが、酸化亜鉛微粒子とグルコン酸を反応させたグルコン酸亜鉛水溶液を用いると、亜鉛イオンとグルコン酸イオンとの相乗作用によって、銀に劣らない抗菌力・殺菌力を得られる。 Here, a case where gluconic acid is used as the organic acid will be described as an example. Part of zinc oxide reacts with gluconic acid in an aqueous solution of gluconic acid to become zinc gluconate (gluconate ion and zinc ion), which becomes soluble in water. Zinc ion is said to have weaker bactericidal activity than silver ion, but when a zinc gluconate aqueous solution obtained by reacting zinc oxide fine particles with gluconic acid is used, it is inferior to silver due to the synergistic action of zinc ion and gluconate ion. No antibacterial and bactericidal power can be obtained.
［化１］に示すように、グルコン酸は水に溶解させると、その一部がグルコノラクトンになる。これを平衡という。グルコノラクトンとグルコン酸の割合は、温度、濃度、ｐＨなどによって変わるが、酸性にすると水中の酸が多くなるので、グルコノラクトンの割合が多くなる。アルカリ性にするとグルコン酸は塩になって安定化するので、グルコノラクトンの割合は少なくなる。平衡とは２つの水槽をチューブで連結したようなもので、左側の水槽に水を入れると、バランスを保つために水はチューブを通って右側に流れ、平衡になる。 As shown in [Chemical formula 1], when gluconic acid is dissolved in water, a part of it becomes gluconolactone. This is called equilibrium. The ratio of gluconolactone and gluconic acid varies depending on the temperature, concentration, pH, etc., but when acidified, the amount of acid in water increases, so the ratio of gluconolactone increases. When made alkaline, gluconic acid becomes a salt and stabilizes, so that the proportion of gluconolactone decreases. Equilibrium is like connecting two tanks with a tube. When water is put into the tank on the left side, water flows to the right side through the tube to maintain balance and becomes equilibrium.
酸化亜鉛ナノ粒子とグルコン酸の相乗効果について、さらに説明する。ウイルスは、カプシドというタンパク質の殻を有している。［化２］に示すように、タンパク質には酸性のアミノ酸と塩基性のアミノ酸が含まれているが、酸性のアミノ酸は、－ＣＯＯＨなどを持っており、マイナスイオン（ＣＯＯ－）になる。塩基性のアミノ酸は、－ＮＨ２を持っているので、プラスイオン（－ＮＨ３ ＋）になる。 The synergistic effect of zinc oxide nanoparticles and gluconic acid will be further described. The virus has a protein shell called capsid. As shown in [Chemical formula 2], proteins contain acidic amino acids and basic amino acids, but acidic amino acids have -COOH and the like and become negative ions ( COO- ). Since the basic amino acid has -NH 2 , it becomes a positive ion (-NH 3+ ).
ここで、タンパク質中のプラスイオンが多かったり、マイナスイオンが多かったりするとタンパク質は水に溶け、マイナスとプラスのイオンが同じ（等電点）だと中性になり、電荷を持たないので水に溶けなくなる。タンパク質を構成要素とするウイルスを等電点にすると、ウイルスは水に溶けなくなり、ウイルス同士が凝集し合って不活性化すると考えられる。 Here, if there are many positive ions or many negative ions in the protein, the protein dissolves in water, and if the negative and positive ions are the same (isoelectric point), it becomes neutral and has no charge, so it is in water. It doesn't melt. It is considered that when a virus whose component is a protein is set to the isoelectric point, the virus becomes insoluble in water and the viruses aggregate and inactivate each other.
酸化亜鉛ナノ粒子とグルコン酸（水に溶解させると一部がラクトン化する有機酸）とを反応させるとグルコン酸亜鉛が得られ、これが上記等電点の調整を行い、ウイルス相互を凝集させるとともに、ウイルスのエンベロープを破壊させるので、ウイルスが不活性化する。 Reacting zinc oxide nanoparticles with gluconic acid (an organic acid that partially lactones when dissolved in water) yields zinc gluconate, which adjusts the isoelectric point and aggregates the viruses together. , The virus is inactivated because it destroys the virus's envelope.
有機酸亜鉛の活性は、溶液中においては５０００～１００００ｐｐｍ含まれる場合に抗ウイルス活性が高い。抗ウイルス活性の観点から、好ましくは６０００～９０００ｐｐｍ、より好ましくは６５００～８５００ｐｐｍとする。 The activity of zinc organic acid is high in antiviral activity when it is contained in a solution at 5000 to 10000 ppm. From the viewpoint of antiviral activity, it is preferably 6000 to 9000 ppm, more preferably 6500 to 8500 ppm.
また、柑橘類種子抽出物の濃度は、好ましくは３０００～９０００ｐｐｍ、より好ましくは４０００～７０００ｐｐｍである。 The concentration of the citrus seed extract is preferably 3000 to 9000 ppm, more preferably 4000 to 7000 ppm.
また、有機酸を含ませることがない場合、酸化亜鉛ナノ粒子の濃度は、好ましくは２０００～２００００ｐｐｍ、より好ましくは３０００～１６０００ｐｐｍ、さらに好ましくは４０００～１２０００ｐｐｍである。 When the organic acid is not contained, the concentration of the zinc oxide nanoparticles is preferably 2000 to 20000 ppm, more preferably 3000 to 16000 ppm, still more preferably 4000 to 12000 ppm.
なお、１，３ジメチル－２－イミダゾリジンを有機酸亜鉛水溶液に添加すると、有機酸亜鉛の水への溶解度を増加させることができる。ここで、１，３ジメチル－２－イミダゾリジンは毒性を有しない点で好適である。 By adding 1,3dimethyl-2-imidazolidine to the aqueous solution of zinc organic acid, the solubility of zinc organic acid in water can be increased. Here, 1,3dimethyl-2-imidazolidine is preferable in that it is not toxic.
ここで柑橘類とは、ミカン科のミカン属、キンカン属、カラタチ属、クリメニア属、エレモシトラス属、ミクロシトラス属からなる。例えば、ウンシュウミカン、グレープフルーツ、レモン、カボス、ライム、ナツミカン、ダイダイ、キンカン、ユズ等が含まれる。本発明では柑橘類について特に限定する必要はないが、中でもグレープフルーツ、ハッサク、ナツミカン、レモン、カボスが好ましく、グレープフルーツが最も好ましい。 Here, citrus fruits are composed of the genus Citrus, the genus Kumquat, the genus Trifoliate orange, the genus Crimenia, the genus Elemocitrus, and the genus Microcitrus of the Rutaceae family. For example, unshu mikan, grapefruit, lemon, kabosu, lime, nut mikan, daidai, kumquat, yuzu and the like are included. In the present invention, citrus fruits are not particularly limited, but grapefruit, hassaku, nutsumikan, lemon, and kabosu are preferable, and grapefruit is most preferable.
柑橘類の種子の抽出に用いる溶媒としては特に限定されることはなく、公知の抽出溶媒を適宜用いることができる。例えば、水、エタノール等のアルコール、ケトン、エーテル、飽和炭化水素、エステル、芳香族化合物などを、１種ないし２種以上混合して使用できる。また、異なる溶媒を用いて複数回の抽出を行ってもよい。 The solvent used for extracting citrus seeds is not particularly limited, and a known extraction solvent can be appropriately used. For example, water, alcohols such as ethanol, ketones, ethers, saturated hydrocarbons, esters, aromatic compounds and the like can be used by mixing one or more. Moreover, you may perform a plurality of extractions using different solvents.
また、柑橘類の種子は、抽出を行いやすくするために、粉砕等の前処理が行われていてもよい。 Further, the citrus seeds may be subjected to pretreatment such as crushing in order to facilitate extraction.
また、抽出温度などの抽出雰囲気は特に限定されず、公知の抽出雰囲気で行えばよい。 Further, the extraction atmosphere such as the extraction temperature is not particularly limited, and the extraction may be performed in a known extraction atmosphere.
本発明にかかる感染症予防薬剤によると、酸化亜鉛微粒子と柑橘類種子抽出物とが協働して、長期間にわたってウイルスの不活性化や抗菌を行うことができる。 According to the infectious disease preventive agent according to the present invention, the zinc oxide fine particles and the citrus seed extract can cooperate with each other to inactivate the virus and perform antibacterial activity for a long period of time.
本発明にかかる抗菌・抗ウイルス薬剤は、酸化亜鉛ナノ粒子と、２５００～１００００ｐｐｍの柑橘類種子抽出物とが、水系の溶媒に分散されてなる。 The antibacterial / antiviral agent according to the present invention comprises zinc oxide nanoparticles and 2500 to 10000 ppm of citrus seed extract dispersed in an aqueous solvent.
水系の溶媒は、グルコン酸、酢酸、クエン酸、酒石酸、リンゴ酸、乳酸等の有機酸を含んでいてもよく、中でも水に溶解させると一部ラクトン化する有機酸（γ－ヒドロキシ酪酸、グルコン酸など）を含んでいることが好ましい。 The aqueous solvent may contain organic acids such as gluconic acid, acetic acid, citric acid, tartrate acid, malic acid, and lactic acid, and among them, organic acids (γ-hydroxybutyric acid, glucon) that partially become lactone when dissolved in water. It is preferable that it contains an acid or the like).
本発明によれば、酸化亜鉛を超微粒子（ナノ粒子）にすることにより、銀に劣らない抗菌性が得られる。 According to the present invention, by making zinc oxide into ultrafine particles (nanoparticles), antibacterial properties comparable to silver can be obtained.
また、グルコン酸などの水に溶解させると一部ラクトン化する有機酸を含んでいる場合には、酸化亜鉛ナノ粒子の一部がグルコン酸と反応してグルコン酸亜鉛となり、酸化亜鉛ナノ粒子がグルコン酸（水に溶解させると一部がラクトン化する有機酸）と反応し、グルコン酸亜鉛となり、これがウイルスの等電点の調整をうまく行ない、またウイルスのエンベロープを破壊させ、これを不活性化させる。 In addition, when it contains an organic acid such as gluconic acid that is partially lactonized when dissolved in water, some of the zinc oxide nanoparticles react with gluconic acid to become zinc gluconate, and the zinc oxide nanoparticles become zinc oxide. It reacts with gluconic acid (an organic acid that partially lactones when dissolved in water) to form zinc gluconate, which successfully regulates the isoelectric point of the virus and also destroys the envelope of the virus, making it inactive. To make it.
また、柑橘類種子抽出物は、抗菌、抗ウイルス効果を有することが知られている。しかしながら、柑橘類種子抽出物の抗菌・抗ウイルス活性は比較的短時間（およそ５～３０日）でなくなってしまう。 In addition, citrus seed extract is known to have antibacterial and antiviral effects. However, the antibacterial and antiviral activity of the citrus seed extract disappears in a relatively short time (about 5 to 30 days).
本発明では、酸化亜鉛ナノ粒子と、柑橘類種子抽出物とを組み合わせて用いているが、この構成によると、酸化亜鉛ナノ粒子単独、柑橘類種子抽出物単独よりも高い抗菌、抗ウイルス効果が、１月以上の長期にわたって得られるようになる。 In the present invention, zinc oxide nanoparticles and citrus seed extract are used in combination. According to this configuration, zinc oxide nanoparticles alone and citrus seed extract alone have higher antibacterial and antiviral effects. It will be available for a long period of more than a month.
抗菌・抗ウイルス薬剤は、例えば市販の柑橘類種子抽出物の溶液と、酸化亜鉛ナノ粒子とを混合し、適宜希釈することにより製造できる。ここで、グルコン酸を含んだ溶液に酸化亜鉛ナノ粒子を混合する場合、次のように行うことが好ましい。 The antibacterial / antiviral agent can be produced, for example, by mixing a solution of a commercially available citrus seed extract with zinc oxide nanoparticles and appropriately diluting the mixture. Here, when the zinc oxide nanoparticles are mixed with the solution containing gluconic acid, it is preferable to carry out as follows.
グルコン酸を４０～５０重量％含むグルコン酸水溶液に、酸化亜鉛ナノ粒子を、得られるグルコン酸亜鉛の濃度が５０００～１００００ｐｐｍになるように、室温で、温度を上昇させないように、かき混ぜて分散させる。グルコン酸水溶液に酸化亜鉛ナノ粒子を分散させていくと、発熱し、温度が上がり過ぎるとゲル化することが認められた。 Zinc oxide nanoparticles are dispersed in a gluconic acid aqueous solution containing 40 to 50% by weight of gluconic acid by stirring at room temperature so that the concentration of the obtained zinc gluconate is 5000 to 10000 ppm and not raising the temperature. .. It was found that when zinc oxide nanoparticles were dispersed in an aqueous solution of gluconic acid, heat was generated and gelation occurred when the temperature was too high.
（実施例１）
直径５０ｎｍ～７０ｎｍの酸化亜鉛ナノ粒子と、グレープフルーツ種子抽出物溶液（株式会社アデプト製Desfan-10）と、水と、を混合し分散させて、実施例１にかかる抗菌・抗ウイルス薬剤を作成した。この薬剤の酸化亜鉛ナノ粒子の濃度は１００００ｐｐｍ、グレープフルーツ種子抽出物の濃度は５０００ｐｐｍであった。なお、Desfan-10は、水９０．５％、グリセリン６．２％、グレープフルーツ種子のエタノール抽出物３．３％により構成されているものである。
(Example 1)
Zinc oxide nanoparticles having a diameter of 50 nm to 70 nm, a grapefruit seed extract solution (Desfan-10 manufactured by Adept Co., Ltd.), and water were mixed and dispersed to prepare an antibacterial / antiviral agent according to Example 1. .. The concentration of zinc oxide nanoparticles of this drug was 10,000 ppm, and the concentration of grapefruit seed extract was 5000 ppm. Desfan-10 is composed of 90.5% water, 6.2% glycerin, and 3.3% ethanol extract of grapefruit seeds.
（実施例２）
グルコン酸５０％水溶液に、直径５０ｎｍ～７０ｎｍの酸化亜鉛ナノ粒子を室温で、温度を上げないように、かき混ぜて分散させた。この後、グレープフルーツ種子抽出物溶液（株式会社アデプト製Desfan-10）と、水と、をさらに加えて、実施例２にかかる抗菌・抗ウイルス薬剤を作成した。この薬剤のグルコン酸亜鉛の濃度は７５００ｐｐｍ、グレープフルーツ種子抽出物の濃度は５０００ｐｐｍであった。
(Example 2)
Zinc oxide nanoparticles having a diameter of 50 nm to 70 nm were dispersed in a 50% aqueous solution of gluconic acid at room temperature by stirring so as not to raise the temperature. After that, a grapefruit seed extract solution (Desfan-10 manufactured by Adept Co., Ltd.) and water were further added to prepare an antibacterial / antiviral agent according to Example 2. The concentration of zinc gluconate of this drug was 7500 ppm, and the concentration of grapefruit seed extract was 5000 ppm.
（比較例１）
グレープフルーツ種子抽出物を加えないこと以外は上記実施例１と同様にして、比較例１にかかる抗菌・抗ウイルス薬剤（酸化亜鉛ナノ粒子濃度が１００００ｐｐｍの溶液）を作成した。
(Comparative Example 1)
An antibacterial / antiviral agent (solution having a zinc oxide nanoparticle concentration of 10000 ppm) according to Comparative Example 1 was prepared in the same manner as in Example 1 above except that the grapefruit seed extract was not added.
（比較例２）
グレープフルーツ種子抽出物を加えないこと以外は上記実施例２と同様にして、比較例２にかかる抗菌・抗ウイルス薬剤（グルコン酸亜鉛の濃度７５００ｐｐｍ）を作成した。
(Comparative Example 2)
An antibacterial / antiviral agent (zinc gluconate concentration 7500 ppm) according to Comparative Example 2 was prepared in the same manner as in Example 2 above except that the grapefruit seed extract was not added.
（比較例３）
グレープフルーツ種子抽出物（株式会社アデプト製Desfan-10）を、濃度が５０００ｐｐｍとなるように希釈したものを、比較例３にかかる抗菌・抗ウイルス薬剤とした。
(Comparative Example 3)
Grapefruit seed extract (Desfan-10 manufactured by Adept Co., Ltd.) diluted to a concentration of 5000 ppm was used as an antibacterial / antiviral agent according to Comparative Example 3.
（抗菌試験）
抗菌試験は、ハロー法（JIS L 1902）により行った。まず、市販のセルロース性のガーゼを円形にカットし、これに上記実施例１，２、比較例１～３にかかる抗菌・抗ウイルス薬剤を１．０ｍｌ染み込ませた。試験菌（黄色ぶどう球菌）を含む寒天平板培地の中央部にこのガーゼを置き、３７℃で２４時間培養した。培養後の試料の発育阻止帯（ハロー）の有無により、抗菌性を評価した。なお、実施例１，２、比較例１～３いずれも、抗菌・抗ウイルス薬剤を浸透させて自然乾燥させた後のガーゼ（直後）と、抗菌・抗ウイルス薬剤を浸透後、２５℃、湿度５５％の恒温槽で３０日保存した後のガーゼとを用いて試験を行った。この結果を下記表１に示す。
(Antibacterial test)
The antibacterial test was performed by the halo method (JIS L 1902). First, a commercially available cellulosic gauze was cut into a circle and impregnated with 1.0 ml of the antibacterial / antiviral agent according to Examples 1 and 2 and Comparative Examples 1 to 3. This gauze was placed in the center of an agar plate medium containing a test bacterium (Staphylococcus aureus) and cultured at 37 ° C. for 24 hours. The antibacterial property was evaluated based on the presence or absence of the growth inhibitory zone (halo) of the sample after culturing. In each of Examples 1 and 2 and Comparative Examples 1 to 3, gauze (immediately after) after being infiltrated with an antibacterial / antiviral agent and naturally dried, and 25 ° C. and humidity after being infiltrated with an antibacterial / antiviral agent. The test was carried out using gauze after storing in a 55% constant temperature bath for 30 days. The results are shown in Table 1 below.
ここで、ハローの大きさを目視で確認したところ、直後においては実施例２＞実施例１＞比較例３＞比較例２＞比較例１、３０日後においては実施例２＞実施例１＞比較例２＞比較例１＞比較例３であった。また、実施例２、実施例１、比較例２、比較例１について、直後と３０日後でハローの大きさの違いは目視では確認できなかった。 Here, when the size of the halo was visually confirmed, immediately after that, Example 2> Example 1> Comparative Example 3> Comparative Example 2> Comparative Example 1, and 30 days later, Example 2> Example 1> Comparison. Example 2> Comparative Example 1> Comparative Example 3. Further, regarding Example 2, Example 1, Comparative Example 2, and Comparative Example 1, the difference in halo size immediately after and 30 days later could not be visually confirmed.
比較例３の結果から、グレープフルーツ種子抽出物単独でグルコン酸亜鉛より高い抗菌効果を有するものの、その効果は３０日後にはほぼ失活していることが分かった。また、比較例１～３の結果から、酸化亜鉛ナノ粒子やグルコン酸亜鉛は長期間にわたって高い抗菌効果を有するものの、その効果はグレープフルーツ種子抽出物よりは小さいことが分かった。さらに、比較例１，２の結果から、グルコン酸亜鉛の効果は酸化亜鉛ナノ粒子よりも高いことが分かった。 From the results of Comparative Example 3, it was found that the grapefruit seed extract alone had a higher antibacterial effect than zinc gluconate, but the effect was almost inactivated after 30 days. Further, from the results of Comparative Examples 1 to 3, it was found that zinc oxide nanoparticles and zinc gluconate have a high antibacterial effect for a long period of time, but the effect is smaller than that of grapefruit seed extract. Furthermore, from the results of Comparative Examples 1 and 2, it was found that the effect of zinc gluconate was higher than that of zinc oxide nanoparticles.
これらに対し、グレープフルーツ種子抽出物と酸化亜鉛ナノ粒子とを含んだ実施例１では、比較例１、２よりも高い効果が持続的に得られたことがわかる。これは、酸化亜鉛ナノ粒子がグレープフルーツ種子抽出物を安定させつつ両者の効果が発揮されたためと考えられる。 On the other hand, it can be seen that in Example 1 containing the grapefruit seed extract and zinc oxide nanoparticles, a higher effect than in Comparative Examples 1 and 2 was continuously obtained. It is considered that this is because the zinc oxide nanoparticles exerted both effects while stabilizing the grapefruit seed extract.
また、グルコン酸とグレープフルーツ種子抽出物と酸化亜鉛ナノ粒子とを含んだ実施例２では、実施例１よりも高い効果が持続的に得られたことがわかる。これは、グルコン酸により酸化亜鉛ナノ粒子やグレープフルーツ種子抽出物の抗菌作用がさらに高められたためと考えられる。 Further, it can be seen that in Example 2 containing gluconic acid, grapefruit seed extract and zinc oxide nanoparticles, a higher effect than in Example 1 was continuously obtained. It is considered that this is because the antibacterial action of zinc oxide nanoparticles and grapefruit seed extract was further enhanced by gluconic acid.
（コロナウイルスの不活性化試験）
実施例１，２、比較例１～３にかかる抗菌・抗ウイルス薬剤０．９ｍｌとネココロナウイルス（Feline enteric coronavirus, WSU 79-1683 1.3×105 EID50/0.1ml）０．１ｍｌを加え、ボルテックスで混合し、室温（２０℃）で１０分間反応させた。試験品とウイルスとの混合液を、５倍希釈したＳＣＤＬＰ（Soybean-Casein Digest Broth with Lecithin & Polysorbate 80）で１００倍に希釈し、反応を停止させ、ウイルス感染値測定試料を得た。なお、対照として、ＰＢＳ（Phosphate buffered saline）を用いて同様に試験を行った。
(Coronavirus inactivation test)
Add 0.9 ml of antibacterial / antiviral agent according to Examples 1 and 2 and Comparative Examples 1 to 3 and 0.1 ml of feline coronavirus (Feline enteric coronavirus, WSU 79-1683 1.3 × 10 5 EID 50 / 0.1 ml). The mixture was mixed by vortex and reacted at room temperature (20 ° C.) for 10 minutes. The mixture of the test product and the virus was diluted 100-fold with SCDLP (Soybean-Casein Digest Broth with Lecithin & Polysorbate 80) diluted 5-fold to stop the reaction, and a virus infection value measurement sample was obtained. As a control, PBS (Phosphate buffered saline) was used in the same test.
検査用の全ネコ胎児由来株化細胞を、あらかじめ９６ウェルプレートに播種し、ＣＯ２インキュベータで４日培養した。この後、培養上澄みを除き、１％ウシ胎児血清を含むダルベッコ変法イーグル培地（日水製薬製）に交換した。 All fetus-derived cell lines for inspection were seeded in advance on 96-well plates and cultured in a CO 2 incubator for 4 days. After that, the culture supernatant was removed, and the medium was replaced with Dulbecco's modified Eagle's medium (manufactured by Nissui Pharmaceutical Co., Ltd.) containing 1% fetal bovine serum.
培養液を除いたウェルに、上記のウイルス感染値測定試料、あるいはこれをＰＢＳで１０倍に希釈したものを２５μｌ接種し、３７℃で１時間、ウイルスを細胞に感染させた。１時間後、ウイルス液を除去し、０．２％ウシ胎児血清を含むダルベッコ変法イーグル培地（シグマアルドリッチ製）を各ウェルに０．１ｍｌ加え、ＣＯ２インキュベータで４日培養した。培養後、ウイルスの増殖来より生じた細胞変性効果を顕微鏡で確認し、ウイルス感染価をReed and Munchの方法により算出した。この結果を下記表２に示す。 The wells from which the culture medium had been removed were inoculated with 25 μl of the above virus infection value measurement sample or a 10-fold diluted product thereof with PBS, and the cells were infected with the virus at 37 ° C. for 1 hour. After 1 hour, the virus solution was removed, 0.1 ml of Dulbecco's modified Eagle's medium (manufactured by Sigma-Aldrich) containing 0.2% fetal bovine serum was added to each well, and the cells were cultured in a CO 2 incubator for 4 days. After culturing, the cytopathic effect caused by the growth of the virus was confirmed under a microscope, and the viral infectivity titer was calculated by the method of Reed and Munch. The results are shown in Table 2 below.
この結果から、グルコン酸亜鉛とグレープフルーツ種子抽出物の両方を用いた実施例２では、対照と比べて約９９．９７％（３桁以上）ウイルス感染価を低減できており、ネココロナウイルスの抗ウイルス化に極めて有効であることが分かった。また、抗ウイルスの性能は、ハロー試験と同様に、実施例２＞実施例１＞比較例３＞比較例２＞比較例１であることが分かった。 From this result, in Example 2 using both zinc gluconate and grapefruit seed extract, the virus infectious titer could be reduced by about 99.97% (3 digits or more) as compared with the control, and the anti-feline coronavirus was prevented. It turned out to be extremely effective for viralization. Further, it was found that the antiviral performance was Example 2> Example 1> Comparative Example 3> Comparative Example 2> Comparative Example 1 as in the halo test.
実施例２、比較例２、比較例３にかかる抗菌・抗ウイルス薬剤を適宜希釈して、最小発育阻止濃度測定法Ｉ（抗菌製品協議会）に準拠して、各種の細菌に対する最小発育阻止濃度（グルコン酸亜鉛の濃度）を測定した。この結果、黄色ブドウ球菌(NBRC12732)、大腸菌(NBRC3972) 、大腸菌O157(ATCC43888)、サルモネラ菌(NRBC3313)、肺炎かん菌(ATCC4352)、緑膿菌(NRBC3080)、メチシリン耐性黄色ブドウ球菌(IID1677)に対して、実施例２は比較例２、比較例３よりも低い最小発育阻止濃度（グルコン酸亜鉛又はグレープフルーツ種子抽出物の濃度）であった。 The antibacterial / antiviral agents according to Example 2, Comparative Example 2 and Comparative Example 3 are appropriately diluted to obtain the minimum inhibitory concentration for various bacteria in accordance with the minimum inhibitory concentration measurement method I (Antibacterial Product Council). (Concentration of zinc gluconate) was measured. As a result, against Staphylococcus aureus (NBRC12732), Escherichia coli (NBRC3972), Escherichia coli O157 (ATCC43888), Salmonella (NRBC3313), Pneumonia bacillus (ATCC4352), Pseudomonas aeruginosa (NRBC3080), and methicillin-resistant Staphylococcus aureus (IID1677). In Example 2, the minimum inhibitory concentration (concentration of zinc gluconate or staphylococcus aureus seed extract) was lower than that of Comparative Example 2 and Comparative Example 3.
この結果から、本発明にかかる抗菌・抗ウイルス薬剤は、グラム陰性菌、グラム陽性菌など、さまざまな細菌に対して高い抗菌効果を有することがわかる。この抗菌・抗ウイルス薬剤は、長期間にわたってその効果を持続するため、例えばカーテン、絨毯、壁紙、座椅子、テーブル、家具等、日常で使用する室内や車内等の構成部材やインテリア用品の抗菌・抗ウイルス化に高い効果を発揮できる。 From this result, it can be seen that the antibacterial / antiviral agent according to the present invention has a high antibacterial effect against various bacteria such as gram-negative bacteria and gram-positive bacteria. Since this antibacterial / antiviral agent lasts its effect for a long period of time, it is an antibacterial agent for interior components and interior goods such as curtains, rugs, wallpaper, sitting chairs, tables, furniture, etc. that are used in daily life.・ Highly effective for anti-virus.
上記実施例では、有機酸としてグルコン酸、柑橘類種子抽出物としてグレープフルーツ種子抽出物を使用したものを用いたが、この発明はこれに限られるものでない。 In the above embodiment, gluconic acid is used as the organic acid and grapefruit seed extract is used as the citrus seed extract, but the present invention is not limited thereto.
今回開示された実施例はすべての点で例示であって制限的なものではないと考えられるべきである。本発明の範囲は上記した説明ではなくて特許請求の範囲によって示され、特許請求の範囲と均等の意味および範囲内でのすべての変更が含まれることが意図される。 The examples disclosed this time should be considered to be exemplary and not restrictive in all respects. The scope of the present invention is shown by the scope of claims rather than the above description, and is intended to include all modifications within the meaning and scope equivalent to the scope of claims.
本発明に係る抗菌・抗ウイルス薬剤では、食品にも添加可能な安全な材料が使用されており、しかも長期間にわたって高い活性で抗菌・抗ウイルスを行うことができる。よって、その産業上の利用可能性は大きい。 In the antibacterial / antiviral agent according to the present invention, a safe material that can be added to food is used, and antibacterial / antiviral can be performed with high activity for a long period of time. Therefore, its industrial applicability is great.
Claims (4)
前記柑橘類種子抽出物の濃度が２５００～１００００ｐｐｍである、抗菌・抗ウイルス薬剤。 Zinc oxide nanoparticles and citrus seed extract are antibacterial and antiviral agents dispersed in an aqueous solvent.
An antibacterial / antiviral agent having a concentration of the citrus seed extract of 2500 to 10000 ppm.
請求項１に記載の抗菌・抗ウイルス薬剤。 The aqueous solvent contains an organic acid that partially lactonizes when dissolved in water.
The antibacterial / antiviral agent according to claim 1.
抗菌・抗ウイルス薬剤は、５０００～１００００ｐｐｍのグルコン酸亜鉛を含む、
請求項２に記載の抗菌・抗ウイルス薬剤。 The organic acid is gluconic acid and is
Antibacterial and antiviral agents contain 5000-10000 ppm zinc gluconate,
The antibacterial / antiviral agent according to claim 2.
請求項１、２又は３に記載の抗菌・抗ウイルス薬剤。 The citrus seed extract comprises a grapefruit seed extract.
The antibacterial / antiviral agent according to claim 1, 2 or 3.
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