JPWO2013069780A1 - Virus infection control agent - Google Patents
Virus infection control agent Download PDFInfo
- Publication number
- JPWO2013069780A1 JPWO2013069780A1 JP2013543048A JP2013543048A JPWO2013069780A1 JP WO2013069780 A1 JPWO2013069780 A1 JP WO2013069780A1 JP 2013543048 A JP2013543048 A JP 2013543048A JP 2013543048 A JP2013543048 A JP 2013543048A JP WO2013069780 A1 JPWO2013069780 A1 JP WO2013069780A1
- Authority
- JP
- Japan
- Prior art keywords
- cashew nut
- nut shell
- virus
- heated
- cardanol
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- CPLXHLVBOLITMK-UHFFFAOYSA-N magnesium oxide Inorganic materials [Mg]=O CPLXHLVBOLITMK-UHFFFAOYSA-N 0.000 description 1
- AXZKOIWUVFPNLO-UHFFFAOYSA-N magnesium;oxygen(2-) Chemical compound [O-2].[Mg+2] AXZKOIWUVFPNLO-UHFFFAOYSA-N 0.000 description 1
- FFEARJCKVFRZRR-UHFFFAOYSA-N methionine Chemical compound CSCCC(N)C(O)=O FFEARJCKVFRZRR-UHFFFAOYSA-N 0.000 description 1
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Classifications
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Abstract
非加熱カシューナッツ殻油、加熱カシューナッツ殻油、アナカルド酸、カルダノール、カルドールおよびカシューナッツ殻粉砕物から選択される１つ以上を含有することを特徴とするウイルス感染防除剤。A virus infection control agent comprising one or more selected from non-heated cashew nut shell oil, heated cashew nut shell oil, anacardic acid, cardanol, cardol and cashew nut shell grind.
Description
本発明は、カシューナッツ殻油（ＣＮＳＬ）を含有するウイルス感染防除剤、飼料添加剤、飼料及びこれらを用いた動物の飼育方法に関する。 The present invention relates to a virus infection control agent, a feed additive, a feed, and an animal breeding method using them, which contain cashew nut shell liquid (CNSL).
ウイルスにより引き起こされる病気は、反芻動物（牛、羊、ヤギ）、豚、家禽（鶏、ウズラ、シチメンチョウ、アヒル、ガチョウ、ダチョウ、エミュー、キジ、ホロホロチョウ）、馬、ペット（犬、猫）、ヒトなどに重篤な病害を発生させる病原体で、畜産業界、ペット業界、ヒトに甚大な被害を与えている。インフルエンザウイルスとしては、鳥インフルエンザが甚大な被害を与え、さらにヒト型に変異することによるパンデミックの回避が重要な課題である。インフルエンザウイルスには豚型、犬型、猫型もあり甚大な被害を与えている。ヘルペスウイルスは、反芻動物に牛伝染性鼻気管炎、豚にオーエスキー病、家禽にマレック病を引き起こす病原体であり、ワクチン接種により防除が試みられているが、近年、ワクチン接種の効果が無くなり甚大な被害を与えている。アルテリウイルスは、豚繁殖・呼吸器障害症候群（ＰＲＲＳ）の病原体であるが、有効な防除方法が無く、甚大な被害を与えている。アフトウイルスは反芻動物の口蹄疫の病原体で、一度発症が認められると感染している反芻動物は全て殺処分しなければならないほどの重篤病害である。ニューカッスル病ウイルスは、鶏をはじめ多種の鳥に感染する。特に鶏の感受性が高く、その伝搬性、病原性から重要な疾患であり、養鶏業においてはとりわけ厳重な注意が必要である。牛コロナウイルス病は、牛コロナウイルス感染を原因とするウシの感染症で、下痢を主な症状とする。下痢が長引くと脱水し、代謝性アシドーシスにより衰弱し、時に死亡する。ロタウイルス病は、牛ロタウイルス感染を原因とするウシの感染症で、元気消失、食欲不振、水様性下痢などを示す。豚、ヤギ、馬、犬などにおいても出生直後から幼若期にかけて急性下痢症を起こす。 Diseases caused by viruses include ruminants (cow, sheep, goats), pigs, poultry (chicken, quail, turkey, duck, goose, ostrich, emu, pheasant, guinea fowl), horses, pets (dogs, cats), It is a pathogen that causes serious diseases in humans and so on, and has caused serious damage to the livestock industry, pet industry, and humans. As an influenza virus, it is important to avoid a pandemic caused by avian influenza that causes enormous damage and is mutated into a human form. Influenza viruses are swine, dogs, and cats, causing serious damage. Herpesviruses are pathogens that cause cattle infectious rhinotracheitis in ruminants, Aujeszky's disease in pigs, and Marek's disease in poultry, and have been tried to control by vaccination. Is causing serious damage. Arterivirus is a pathogen of swine reproduction / respiratory disorder syndrome (PRRS), but there is no effective control method, and it causes great damage. Aphthovirus is a pathogen of ruminant foot-and-mouth disease in ruminants, and once a symptom is observed, all infected ruminants are serious diseases that must be killed. Newcastle disease virus infects many types of birds, including chickens. Particularly sensitive to chickens, it is an important disease because of its transmission and pathogenicity. In the poultry industry, particularly strict attention is required. Bovine coronavirus disease is a bovine infection caused by bovine coronavirus infection, and diarrhea is the main symptom. If diarrhea persists, it will dehydrate, weaken by metabolic acidosis, and sometimes die. Rotavirus disease is an infectious disease of cattle caused by bovine rotavirus infection, showing loss of energy, loss of appetite, watery diarrhea and the like. In pigs, goats, horses, dogs, etc., acute diarrhea occurs from birth to early childhood.
特許文献１にはイチョウ抽出液を、レンチウイルスに属するＨＩＶの予防や治療に使用することが記載されている。しかしながら、イチョウから有効成分を抽出する操作は煩雑で大量に得ることは困難である。また、レンチウイルスに対する効果のみ確認されており、他のウイルスに対する効果は不明である。
特許文献２にはカシューナッツ種皮より抽出されたカルドールに、フコシダーゼ阻害活性が有ることが記載されている。しかしながら、ウイルスに対する効果は実際に確認されていない。Patent Document 1 describes that the ginkgo biloba extract is used for prevention and treatment of HIV belonging to lentivirus. However, the operation of extracting an active ingredient from ginkgo is cumbersome and difficult to obtain in large quantities. Moreover, only the effect with respect to a lentivirus is confirmed, and the effect with respect to another virus is unknown.
Patent Document 2 describes that cardol extracted from cashew nut seed coat has fucosidase inhibitory activity. However, the effect on the virus has not been confirmed.
本発明は、従来は予防・治療が困難であったウイルスによる病害を防除するウイルス感染防除剤を提供することを課題とする。 An object of the present invention is to provide a virus infection control agent that controls a disease caused by a virus that has been difficult to prevent or treat.
本発明者らは、上記課題を解決すべく鋭意研究を行った結果、カシューナッツ殻油がインフルエンザウイルスなどのウイルスを不活性化できることを見出した。
本発明者らは、このようにして、本発明を完成するに至った。As a result of intensive studies to solve the above problems, the present inventors have found that cashew nut shell liquid can inactivate viruses such as influenza virus.
The inventors have thus completed the present invention.
すなわち、本発明は以下のとおりである。
（１）非加熱カシューナッツ殻油、加熱カシューナッツ殻油、アナカルド酸、カルダノール、カルドールおよびカシューナッツ殻粉砕物から選択される１つ以上を含有することを特徴とするウイルス感染防除剤。
（２）ウイルスがインフルエンザウイルス、ヘルペスウイルス、アルテリウイルス、アフトウイルス、ニューカッスル病ウイルス、コロナウイルス、ロタウイルス、またはノロウイルスである、（１）記載のウイルス感染防除剤。
（３）（１）または（２）に記載のウイルス感染防除剤を含むことを特徴とする、飼料添加剤。
（４）（３）記載の飼料添加剤を含むことを特徴とする、飼料。
（５）反芻動物、豚、家禽、馬、またはペット用である、（４）記載の飼料。
（６）（４）または（５）に記載の飼料を摂取させることを特徴とする飼育方法。
（７）非加熱カシューナッツ殻油、加熱カシューナッツ殻油、アナカルド酸、カルダノール、カルドールおよびカシューナッツ殻粉砕物から選択される１つ以上を動物に投与する工程を含む、動物におけるウイルス感染防除方法。
（８）動物におけるウイルス感染防除のための、非加熱カシューナッツ殻油、加熱カシューナッツ殻油、アナカルド酸、カルダノール、カルドールおよび／またはカシューナッツ殻粉砕物。That is, the present invention is as follows.
(1) A virus infection control agent comprising one or more selected from non-heated cashew nut shell liquid, heated cashew nut shell oil, anacardic acid, cardanol, cardol and cashew nut shell pulverized product.
(2) The virus infection control agent according to (1), wherein the virus is an influenza virus, herpes virus, arterivirus, aft virus, Newcastle disease virus, coronavirus, rotavirus, or norovirus.
(3) A feed additive comprising the virus infection control agent according to (1) or (2).
(4) A feed comprising the feed additive according to (3).
(5) The feed according to (4), which is for ruminants, pigs, poultry, horses, or pets.
(6) A breeding method characterized by feeding the feed according to (4) or (5).
(7) A method for controlling viral infection in an animal, comprising a step of administering to the animal one or more selected from non-heated cashew nut shell oil, heated cashew nut shell oil, anacardic acid, cardanol, cardol and cashew nut shell pulverized product.
(8) Non-heated cashew nut shell liquid, heated cashew nut shell oil, anacardic acid, cardanol, cardol and / or cashew nut shell pulverized product for controlling virus infection in animals.
本発明のウイルス感染防除剤の原料である、非加熱カシューナッツ殻油、加熱カシューナッツ殻油、およびカシューナッツ殻粉砕物、ならびにそれらの有効成分であるアナカルド酸、カルダノールおよびカルドールは、安価に大量に入手が可能である。
本発明のウイルス感染防除剤は、飼料や飲水に含ませて動物に投与することにより、ウイルス病を効果的に防除することができる。
従って、本発明のウイルス感染防除剤により、安価かつ簡便に、ウイルスを防除することができる。Non-heated cashew nut shell liquid, heated cashew nut shell oil, and cashew nut shell pulverized material, and their active ingredients, anacardic acid, cardanol and cardol, which are raw materials for the virus infection control agent of the present invention, are available in large quantities at low cost. Is possible.
The viral infection control agent of the present invention can effectively control viral diseases by being included in feed or drinking water and administered to animals.
Therefore, the virus infection control agent of the present invention can control the virus at low cost and in a simple manner.
本発明のウイルス感染防除剤は、非加熱カシューナッツ殻油、加熱カシューナッツ殻油、アナカルド酸、カルダノール、カルドールおよびカシューナッツ殻粉砕物から選択される１つ以上を含有することを特徴とする。 The virus infection control agent of the present invention comprises one or more selected from non-heated cashew nut shell liquid, heated cashew nut shell oil, anacardic acid, cardanol, cardol and cashew nut shell pulverized product.
カシューナッツ殻油は、カシューナッツ ツリー(Anacardium occidentale L.)の実の殻に含まれる油状の液体である。カシューナッツ殻油は、その成分として、アナカルド酸、カルダノール、カルドールを含むものである。一般に、アナカルド酸は加熱処理することによりカルダノールに変換される。
カシューナッツの殻を圧搾することにより抽出された非加熱カシューナッツ殻油は、J.Agric.Food Chem. 2001, 49, 2548-2551に記載されるように、アナカルド酸を55〜80質量%、カルダノールを5〜20質量%、カルドールを5〜30質量%含むものである。
非加熱カシューナッツ殻油を130℃以上で加熱処理して得られる加熱カシューナッツ殻油は、非加熱カシューナッツ殻油の主成分のアナカルド酸が脱炭酸しカルダノールに変換され、アナカルド酸を0〜10質量%、カルダノールを55〜80質量%、カルドールを5〜30質量%含むものとなる。Cashew nut shell oil is an oily liquid contained in the shell of the cashew nut tree ( Anacardium occidentale L.). Cashew nut shell oil contains anacardic acid, cardanol, and cardol as its components. In general, anacardic acid is converted to cardanol by heat treatment.
Non-heated cashew nut shell oil extracted by squeezing cashew nut shell contains 55 to 80% by mass of anacardic acid and cardanol as described in J. Agric. Food Chem. 2001, 49, 2548-2551. 5 to 20% by mass and 5 to 30% by mass of cardol.
Heated cashew nut shell oil obtained by heat-treating non-heated cashew nut shell oil at 130 ° C or higher is converted to cardanol by decarboxylation of anacardic acid, the main component of non-heated cashew nut shell oil, and anacardic acid is 0 to 10% by mass , 55 to 80% by mass of cardanol and 5 to 30% by mass of cardol.
カシューナッツ殻油は、カシューナッツの殻を圧搾することにより抽出された植物油として得ることができる。また、カシューナッツ殻油は、抽出により、例えば、カシューナッツ殻を溶剤抽出して得ることもできる。さらに、カシューナッツ殻油は、特開平8-231410号公報に記載されている溶剤抽出法によって得ることができる。
カシューナッツ殻油は、市販品を用いることもできる。
本発明のカシューナッツ殻油は、上記のようにして得られた非加熱カシューナッツ殻油を、70℃以上、好ましくは130℃以上に加熱することによって得た加熱カシューナッツ殻油であってもよい。
本発明で使用するカシューナッツ殻油は、カシューナッツの殻から圧搾抽出(非加熱カシューナッツ殻油)し、これを130℃に加熱処理して得たものでもよい。
本発明で使用するウイルス感染防除剤は、カシューナッツ殻油の代わりに、その有効成分であるアナカルド酸、カルダノールおよび／またはカルドールを含んでいてもよいし、カシューナッツ殻および／またはその粉砕物を含んでいてもよい。Cashew nut shell oil can be obtained as a vegetable oil extracted by pressing the cashew nut shell. Moreover, cashew nut shell liquid can also be obtained by extraction, for example, solvent extraction of cashew nut shell. Further, cashew nut shell liquid can be obtained by a solvent extraction method described in JP-A-8-231410.
A commercial item can also be used for cashew nut shell liquid.
The cashew nut shell liquid of the present invention may be a heated cashew nut shell oil obtained by heating the non-heated cashew nut shell oil obtained as described above to 70 ° C or higher, preferably 130 ° C or higher.
The cashew nut shell oil used in the present invention may be obtained by compressing and extracting cashew nut shell (non-heated cashew nut shell oil) and heat-treating it at 130 ° C.
The virus infection control agent used in the present invention may contain anacardic acid, cardanol and / or cardol, which are its active ingredients, instead of cashew nut shell oil, or cashew nut shell and / or pulverized product thereof. May be.
本発明のウイルス感染防除剤におけるカシューナッツ殻油の含有量は、ウイルス感染防除剤の全量基準で、０．００５質量%〜７０質量%、好ましくは０．０１質量%〜７０質量%、さらに好ましくは０．０２質量%〜７０質量%である。０．００５質量%以上であれば、ウイルスによる病害の防除効果を効率的に奏することができ、７０質量%以下であればウイルス感染防除剤の物性を維持することができるので好ましい。 The content of cashew nut shell oil in the virus infection control agent of the present invention is 0.005% by mass to 70% by mass, preferably 0.01% by mass to 70% by mass, more preferably, based on the total amount of the virus infection control agent. It is 0.02 mass%-70 mass%. If it is 0.005 mass% or more, the effect of controlling diseases caused by viruses can be efficiently achieved, and if it is 70 mass% or less, the physical properties of the virus infection control agent can be maintained, which is preferable.
本発明において使用されるアナカルド酸としては、天然物アナカルド酸、合成アナカルド酸、それらの誘導体が挙げられる。また、市販のアナカルド酸を用いてもよい。アナカルド酸は、特開平8-231410号公報に記載されるように、カシューナッツの殻を有機溶剤で抽出処理して得られたカシューナッツ油を、例えば、シリカゲルカラムクロマトグラフィーを用いてn-ヘキサン、酢酸エチルおよび酢酸の混合溶媒の比率を変えて溶出することによって得ることができる(特開平3-240721号公報、特開平3-240716号公報など)。このようなアナカルド酸は、カシューナッツ殻油と同様の含有量で、ウイルス感染防除剤に含めることができる。 Examples of the anacardic acid used in the present invention include natural products anacardic acid, synthetic anacardic acid, and derivatives thereof. Commercial anacardic acid may also be used. As described in JP-A-8-231410, anacardic acid is obtained by using cashew nut oil obtained by extracting cashew nut shells with an organic solvent, for example, n-hexane, acetic acid using silica gel column chromatography. It can be obtained by changing the ratio of the mixed solvent of ethyl and acetic acid (JP-A-3-240721, JP-A-3-240716, etc.). Such anacardic acid has the same content as cashew nut shell liquid and can be included in the virus infection control agent.
本発明において使用されるカルダノールとしては、天然物カルダノール、合成カルダノール、それらの誘導体が挙げられる。また、本発明において使用されるカルダノールは、カシューナッツ殻油の主成分のアナカルド酸を脱炭酸することにより、得ることができる。このようなカルダノールは、カシューナッツ殻油と同様の含有量で、ウイルス感染防除剤に含めることができる。
なお、加熱カシューナッツ殻油を用いる場合、加熱カシューナッツ殻油中のアナカルド酸とカルダノールとの質量比は、好ましくは、0:100〜20:80である。Examples of cardanol used in the present invention include natural product cardanol, synthetic cardanol, and derivatives thereof. The cardanol used in the present invention can be obtained by decarboxylating anacardic acid, which is the main component of cashew nut shell liquid. Such cardanol has the same content as cashew nut shell liquid and can be included in a virus infection control agent.
In addition, when using heated cashew nut shell liquid, the mass ratio of anacardic acid and cardanol in the heated cashew nut shell oil is preferably 0: 100 to 20:80.
本発明において使用されるカルドールとしては、天然物カルドール、合成カルドール、それらの誘導体が挙げられる。また、本発明において使用されるカルドールは、カシューナッツ殻油から精製することにより、得ることもできる。このようなカルドールは、カシューナッツ殻油と同様の含有量で、ウイルス感染防除剤に含めることができる。 Examples of the cardol used in the present invention include a natural product cardol, a synthetic cardol, and derivatives thereof. The cardol used in the present invention can also be obtained by purification from cashew nut shell liquid. Such cardol can be contained in a virus infection control agent with the same content as cashew nut shell liquid.
本発明においては、非加熱カシューナッツ殻油、加熱カシューナッツ殻油、アナカルド酸、カルダノールおよびカルドールの原液、並びにカシューナッツ殻粉砕物から選択される１つ以上を動物に直接経口投与することもできる。
本発明において使用されるカシューナッツ殻油は、カシューナッツ殻を粉砕・破砕して用いてもよいが、含有しているカシューナッツ殻油（ＣＮＳＬ）に換算して（カシューナッツ殻にはＣＮＳＬが３０％含まれている）同様の含有量とすればよい。In the present invention, one or more selected from unheated cashew nut shell liquid, heated cashew nut shell oil, anacardic acid, cardanol and cardol stock solution, and cashew nut shell grind can be directly orally administered to the animal.
The cashew nut shell oil used in the present invention may be used by pulverizing and crushing the cashew nut shell, but converted to the cashew nut shell oil (CNSL) contained therein (the cashew nut shell contains 30% CNSL). The content may be the same.
本発明の防除剤は、ウイルス、好ましくは、インフルエンザウイルス、ヘルペスウイルス、アルテリウイルス、アフトウイルス、ニューカッスル病ウイルス、コロナウイルス、ロタウイルス、またはノロウイルスを防除するために使用される。なお、アルテリウイルス科に属する豚繁殖・呼吸器障害症候群（ＰＲＲＳ）ウイルスはアルテリウイルスに含まれる。
本発明において、防除とは、ウイルス病の予防および治療を含み、具体的にはウイルスの不活化および除去を含む。ここで、ウイルスの不活化とは、ウイルスの活性や伝染力を不活化する行為をいう。ウイルスの不活化は、例えば、ウイルス感染による細胞の形態変化の非存在を観察することにより、確認することができる。The control agent of the present invention is used for controlling viruses, preferably influenza viruses, herpes viruses, arteriviruses, aphthviruses, Newcastle disease viruses, coronaviruses, rotaviruses, or noroviruses. In addition, the porcine reproduction / respiratory disorder syndrome (PRRS) virus belonging to the Arteriviridae family is included in the Arterivirus.
In the present invention, the term “control” includes prevention and treatment of viral diseases, and specifically includes inactivation and removal of viruses. Here, virus inactivation refers to an act of inactivating virus activity and infectivity. Inactivation of the virus can be confirmed, for example, by observing the absence of cell shape change due to virus infection.
本発明のウイルス感染防除剤として、医薬製剤の製造法で一般的に用いられている公知の手段に従って、非加熱カシューナッツ殻油、加熱カシューナッツ殻油、アナカルド酸、カルダノール、カルドールおよびカシューナッツ殻粉砕物から選択される１つ以上を、そのまま、あるいは薬理学的に許容される担体と混合してヒトを含む動物に投与することができる。
薬理学的に許容される担体としては、例えば固形製剤における賦形剤、滑沢剤、結合剤及び崩壊剤、あるいは液状製剤における溶剤、溶解補助剤、懸濁化剤、等張化剤、緩衝剤及び無痛化剤等が挙げられる。
本発明のウイルス感染防除剤の製剤化には、通常製剤化に用いられる各種の成分が任意に使用されるが、その例としては、例えばデンプン、デキストリン、乳糖、コーンスターチ、無機塩類などが挙げられる。As a virus infection control agent of the present invention, from unheated cashew nut shell oil, heated cashew nut shell oil, anacardic acid, cardanol, cardol and cashew nut shell ground material according to known means generally used in the production method of pharmaceutical preparations One or more selected can be administered to animals including humans as it is or mixed with a pharmacologically acceptable carrier.
Examples of pharmacologically acceptable carriers include excipients, lubricants, binders and disintegrants in solid formulations, or solvents, solubilizers, suspending agents, isotonic agents, buffers in liquid formulations. And soothing agents.
For the formulation of the virus infection control agent of the present invention, various components usually used for formulation are optionally used. Examples thereof include starch, dextrin, lactose, corn starch, and inorganic salts. .
本発明のウイルス感染防除剤の剤型としては、アンプル、錠剤、カプセル剤、顆粒剤、細粒剤、散剤、輸液、ドリンク剤等が挙げられるが、特定の剤型のものに限定されるものではない。
剤形の形態によっては、前記したカシューナッツ殻の粉砕・破砕物、もしくは何らの処理もしていないカシューナッツ殻ならびに／または非加熱カシューナッツ殻油、加熱カシューナッツ殻油、アナカルド酸、カルダノールおよびカルドールから選択される１つ以上をそのまま他の任意成分と混合させて本発明のウイルス感染防除剤とすることができる。また、他の任意成分と混合させず、粉砕・破砕物そのものもしくはカシューナッツ殻そのものならびに／または非加熱カシューナッツ殻油、加熱カシューナッツ殻油、アナカルド酸、カルダノールおよびカルドールから選択される１つ以上のみを、飼料添加剤、さらには飼料とすることもできる。さらに、非加熱カシューナッツ殻油、加熱カシューナッツ殻油、アナカルド酸、カルダノールおよびカルドールから選択される１つ以上をエタノールなどの溶媒に溶解させ、これを、飼料用組成物または飼料に混合吸収させることもできる。Examples of the dosage form of the virus infection control agent of the present invention include ampoules, tablets, capsules, granules, fine granules, powders, infusions, drinks, etc., but are limited to specific dosage forms. is not.
Depending on the form of the dosage form, it is selected from the above-mentioned crushed and crushed cashew nut shells or untreated cashew nut shells and / or unheated cashew nut shell oil, heated cashew nut shell oil, anacardic acid, cardanol and cardol One or more can be directly mixed with other optional components to obtain the virus infection control agent of the present invention. Moreover, without mixing with other optional components, only one or more selected from crushed and crushed material or cashew nut shell itself and / or unheated cashew nut shell oil, heated cashew nut shell oil, anacardic acid, cardanol and cardol, It can also be used as a feed additive or even a feed. Further, one or more selected from unheated cashew nut shell liquid, heated cashew nut shell liquid, anacardic acid, cardanol and cardol may be dissolved in a solvent such as ethanol, and this may be mixed and absorbed in the feed composition or feed. it can.
本発明のウイルス感染防除剤のヒトに対する投与方法としては特に制限されず、経口投与、静脈投与などが挙げられる。好ましい投与量は、投与対象、対象臓器、症状、投与方法などにより異なり特に制限されないが、例えば、患者（体重６０ｋｇとして）に対して、一日につき約０．０１〜２３０ｇ、好ましくは約０．０２５〜２４０ｇ、より好ましくは約０．１〜２５０ｇである。 The method for administering the viral infection control agent of the present invention to humans is not particularly limited, and examples thereof include oral administration and intravenous administration. The preferred dose varies depending on the administration subject, target organ, symptom, administration method and the like, and is not particularly limited. For example, for a patient (weight 60 kg), about 0.01 to 230 g per day, preferably about 0. 025 to 240 g, more preferably about 0.1 to 250 g.
本発明の飼料添加剤は牛、豚、鶏などの家畜用飼料、ペットフード、ペット用サプリメント（以下、飼料という。）に用いられる他の飼料成分と混合して、本発明の飼料とすることができる。飼料の種類や、非加熱カシューナッツ殻油、加熱カシューナッツ殻油、アナカルド酸、カルダノール、カルドールおよびカシューナッツ殻粉砕物以外の成分は、特に制限されない。好ましい含有量は、飼料の乾物質量あたり、全量基準で０．００１質量％〜７０質量％が好ましく、より好ましくは０．００５質量％〜７０質量％、特に好ましくは０．０１質量％〜７０質量％である。０．００１質量％以上であれば、ウイルス感染防除効果が向上することが期待できる。 The feed additive of the present invention is mixed with other feed ingredients used in livestock feed such as cattle, pigs and chickens, pet food, and supplements for pets (hereinafter referred to as feed) to obtain the feed of the present invention. Can do. There are no particular limitations on the type of feed and ingredients other than unheated cashew nut shell liquid, heated cashew nut shell oil, anacardic acid, cardanol, cardol and cashew nut shell grind. The content is preferably 0.001% by mass to 70% by mass, more preferably 0.005% by mass to 70% by mass, and particularly preferably 0.01% by mass to 70% by mass based on the total amount of dry matter in the feed. %. If it is 0.001 mass% or more, it can be expected that the virus infection control effect is improved.
本発明の飼料は、非加熱カシューナッツ殻油、加熱カシューナッツ殻油、アナカルド酸、カルダノール、カルドールおよびカシューナッツ殻粉砕物から選択される１つ以上を含有することを特徴とする。
なお、本発明の飼料におけるカシューナッツ殻油、加熱処理カシューナッツ殻油、アナカルド酸、カルダノール、カルドールおよびカシューナッツ殻粉砕物の含有量は、飼料の乾物質量当たり、全量基準で０．００１質量％〜２．０質量％が好ましく、より好ましくは０．００５質量％〜１．０質量％、特に好ましくは０．０１質量％〜０．５質量％である。０．００１質量％以上であれば、ウイルス感染防除効果が向上することが期待できる。一方、動物の嗜好性改善のためには、２．０質量％以下が好ましい。The feed of the present invention contains one or more selected from non-heated cashew nut shell liquid, heated cashew nut shell oil, anacardic acid, cardanol, cardol, and cashew nut shell pulverized product.
In addition, the content of cashew nut shell liquid, heat-treated cashew nut shell oil, anacardic acid, cardanol, cardol, and cashew nut shell ground material in the feed of the present invention is 0.001% by mass to 2.% by weight based on the total amount of dry matter in the feed. 0 mass% is preferable, More preferably, it is 0.005 mass%-1.0 mass%, Most preferably, it is 0.01 mass%-0.5 mass%. If it is 0.001 mass% or more, it can be expected that the virus infection control effect is improved. On the other hand, it is preferably 2.0% by mass or less for improving the palatability of animals.
本発明の飼料は、飼料添加剤をそのまま飼料成分に添加し、混合して製造することができる。この際、粉末状、固形状の飼料添加剤を用いる場合は、混合を容易にするために飼料添加剤を液状又はゲル状の形態にしてもよい。この場合は、水、大豆油、菜種油、コーン油などの植物油、液体動物油、ポリビニルアルコールやポリビニルピロリドン、ポリアクリル酸などの水溶性高分子化合物を液体担体として用いることができる。また、飼料中におけるカシューナッツ殻油の均一性を保つために、アルギン酸、アルギン酸ナトリウム、キサンタンガム、カゼインナトリウム、アラビアゴム、グアーガム、タマリンド種子多糖類などの水溶性多糖類を配合することも好ましい。 The feed of the present invention can be produced by adding a feed additive to a feed component as it is and mixing them. At this time, when a powdered or solid feed additive is used, the feed additive may be in a liquid or gel form for easy mixing. In this case, water, vegetable oils such as soybean oil, rapeseed oil and corn oil, liquid animal oils, water-soluble polymer compounds such as polyvinyl alcohol, polyvinyl pyrrolidone and polyacrylic acid can be used as the liquid carrier. In order to maintain the uniformity of the cashew nut shell liquid in the feed, it is also preferable to add a water-soluble polysaccharide such as alginic acid, sodium alginate, xanthan gum, sodium caseinate, gum arabic, guar gum, tamarind seed polysaccharide.
本発明の飼料は、非加熱カシューナッツ殻油、加熱カシューナッツ殻油、アナカルド酸、カルダノール、カルドールおよびカシューナッツ殻粉砕物から選択される１つ以上に加えて、家畜動物の成長促進に有効な成分、栄養補助成分、保存安定性を高める成分等の任意成分をさらに含むものであってもよい。このような任意成分としては、例えば、エンテロコッカス属細菌、バチルス属細菌、ビフィズス菌、ラクトバチルス属細菌等の生菌剤；アミラーゼ、リパーゼ等の酵素；Ｌ−アスコルビン酸、塩化コリン、イノシトール、葉酸等のビタミン；塩化カリウム、クエン酸鉄、酸化マグネシウム、リン酸塩類等のミネラル；ＤＬ−アラニン、ＤＬ−メチオニン、塩酸Ｌ−リジン等のアミノ酸；フマル酸、酪酸、乳酸、酢酸等の有機酸及びそれらの塩；エトキシキン、ジブチルヒドロキシトルエン等の抗酸化剤；プロピオン酸カルシウム等の防カビ剤；カルボキシルメチルセルロース（ＣＭＣ）、カゼインナトリウム、ポリアクリル酸ナトリウム等の粘結剤；グリセリン脂肪酸エステル、ソルビタン脂肪酸エステル等の乳化剤；アスタキサンチン、カンタキサンチン等の色素；各種エステル、エーテル、ケトン類等の着香料、が挙げられる。 The feed according to the present invention comprises, in addition to one or more selected from non-heated cashew nut shell oil, heated cashew nut shell oil, anacardic acid, cardanol, cardol and cashew nut shell ground material, an ingredient effective for promoting the growth of livestock animals, nutrition It may further contain an optional component such as an auxiliary component or a component that enhances storage stability. Examples of such optional components include live bacteria such as Enterococcus bacteria, Bacillus bacteria, Bifidobacteria, and Lactobacillus bacteria; Amylase, lipase and other enzymes; L-ascorbic acid, choline chloride, inositol, folic acid, and the like Vitamins; minerals such as potassium chloride, iron citrate, magnesium oxide, phosphates; amino acids such as DL-alanine, DL-methionine, L-lysine hydrochloride; organic acids such as fumaric acid, butyric acid, lactic acid, acetic acid and the like Antioxidants such as ethoxyquin and dibutylhydroxytoluene; Antifungal agents such as calcium propionate; Binders such as carboxymethylcellulose (CMC), sodium caseinate and sodium polyacrylate; Glycerin fatty acid ester, sorbitan fatty acid ester, etc. Emulsifiers; astaxanthin, mosquito Dyes data xanthine and the like; various esters, ethers, flavoring such as ketones, and the like.
本発明のウイルス感染防除剤を投与する動物の種類は、上述したようなウイルスに感染しうる動物であれば特に制限はされないが、哺乳類又は鳥類、例えば反芻動物（牛、羊、ヤギ）、豚、家禽（鶏、ウズラ、シチメンチョウ、アヒル、ガチョウ、ダチョウ、エミュー、キジ、ホロホロチョウ）、馬、ペット（犬、猫）である。摂取させる飼料の量は、動物の種類、体重、年齢、性別、健康状態、増体促進飼料の成分などにより適宜調節することができ、このとき本発明の飼料に含まれる非加熱カシューナッツ殻油、加熱カシューナッツ殻油、アナカルド酸、カルダノール、カルドールおよびカシューナッツ殻粉砕物から選択される１つ以上は、好ましくは０．０１〜１００ｇ／体重１００ｋｇ・日、より好ましくは０．０２５〜１００ｇ／体重１００ｋｇ・日、さらに好ましくは０．１〜１００ｇ／体重１００ｋｇ・日である。
本発明の飼料を摂取させる方法及び飼育する方法は、動物の種類に応じて、通常用いられる方法をとることができる。The type of animal to which the virus infection control agent of the present invention is administered is not particularly limited as long as it is an animal that can be infected with the virus as described above, but mammals or birds such as ruminants (cow, sheep, goat), pigs. , Poultry (chicken, quail, turkey, duck, goose, ostrich, emu, pheasant, guinea fowl), horse, pet (dog, cat). The amount of feed to be ingested can be appropriately adjusted according to the type of animal, body weight, age, sex, health condition, ingredients of the body weight-enhancing feed, etc., at this time, unheated cashew nut shell liquid contained in the feed of the present invention, One or more selected from heated cashew nut shell oil, anacardic acid, cardanol, cardol and cashew nut shell grind is preferably 0.01-100 g / 100 kg body weight / day, more preferably 0.025-100 g / 100 kg body weight / day. Day, more preferably 0.1 to 100 g / body weight 100 kg · day.
The method of ingesting the feed of the present invention and the method of rearing it can be a commonly used method depending on the type of animal.
本発明のウイルス感染防除剤の適用方法としては、特に制限はないが、例えば塗料に配合して塗布材料としたり、樹脂材料に配合して成形したり、あらかじめ形成した成形体に塗料などの塗布材料と共に塗布するなどして、樹脂成形体に備えるようにした抗ウイルス性樹脂成形体とするなど、任意の方法により適用することが出来る。また、これらの施設・環境用途に供する場合でも、非加熱カシューナッツ殻油、加熱カシューナッツ殻油、アナカルド酸、カルダノール、カルドールおよびカシューナッツ殻粉砕物から選択される１つ以上を、そのまま、あるいは各用途に許容される担体と混合して使用することができる。その他必要に応じて、ウイルス感染防除剤の酸化防止剤、安定化剤、防腐剤と併用することができる。
このような抗ウイルス性を備えた樹脂成形体や塗布材料は、ウイルスが侵入しやすい環境に使われる部品、具体的には換気フィルターやマスク等における化学繊維、グラスペーパー及び活性アルミナ、あるいは施設洗浄剤における溶剤、溶解補助剤、懸濁化剤、等張化剤、緩衝剤、あるいは施設塗料における塗膜形成成分、添加剤、溶剤、顔料等に広く適用することができる。更には、高度な衛生管理が求められる環境で使用される繊維製品や樹脂製品、化学薬品、具体的にはエプロン、布巾、カーテン、コーティング剤、シーリング材、クーリングタワー用防藻剤、ワックス等へも適用することができる。
なお、本発明の剤を利用した塗料は、ウイルス感染防除用途に限らず、抗菌塗料として使用することもできる。The method for applying the virus infection control agent of the present invention is not particularly limited, but for example, it is blended into a paint to form a coating material, blended with a resin material and molded, or coated with a preform on a molded body. It can be applied by any method, for example, by applying it together with the material to obtain an antiviral resin molded body prepared for the resin molded body. In addition, even when used for these facilities and environmental uses, one or more selected from unheated cashew nut shell oil, heated cashew nut shell oil, anacardic acid, cardanol, cardol, and cashew nut shell pulverized product may be used as they are or for each use. It can be used in admixture with an acceptable carrier. In addition, it can be used in combination with antioxidants, stabilizers, and preservatives of virus infection control agents as necessary.
Resin moldings and coating materials with such antiviral properties are components used in environments where viruses are likely to enter, specifically chemical fibers in ventilation filters and masks, glass paper and activated alumina, or facility cleaning. It can be widely applied to solvents, solubilizing agents, suspending agents, tonicity agents, buffering agents, coating film forming components, additives, solvents, pigments, etc. in facility paints. In addition, textile products and resin products used in environments where advanced hygiene management is required, chemicals, specifically apron, cloth, curtains, coating agents, sealing materials, cooling tower algae, wax, etc. Can be applied.
In addition, the coating material using the agent of the present invention can be used not only as a virus infection control application but also as an antibacterial coating material.
なお、以上説明した態様は、本発明の一態様を示したものであって、本発明は、前記した実施形態に限定されるものではなく、本発明の目的および効果を達成できる範囲内での変形や改良が、本発明の内容に含まれるものであることは言うまでもない。また、本発明を実施する際における具体的な構造及び形状などは、本発明の目的および効果を達成できる範囲内において、他の構造や形状などとしても問題はない。 The aspect described above shows one aspect of the present invention, and the present invention is not limited to the above-described embodiment, and is within a range in which the object and effect of the present invention can be achieved. It goes without saying that modifications and improvements are included in the content of the present invention. Further, the specific structure and shape in carrying out the present invention are not problematic as other structures and shapes within the scope of achieving the object and effect of the present invention.
製造例
カシュー・トレーディング（株）よりカシューナッツの殻５００ｋｇを入手し、圧搾することによりカシューナッツ殻油（非加熱ＣＮＳＬ）１５８ｋｇを製造した。また、１３０℃で加熱処理することによりアナカルド酸をカルダノールに変換した加熱処理カシューナッツ殻油（加熱ＣＮＳＬ）を、カシュー・トレーディング（株）より入手した（コールドプレスオイル（インド産））。
ＣＮＳＬの組成は以下の方法で測定した。すなわち、HPLC（Waters600、日本ウォーターズ（株））、検出機（Waters490E、日本ウォーターズ（株））、プリンタ(クロマトパックＣ−Ｒ６Ａ、島津製作所)、カラム（SUPELCOSIL LC18、SUPELCO社）を用いた。アセトニトリル：水：酢酸が８０：２０：１（容量比）の溶媒を用い、流速は２ｍｌ／分とした。２８０ｎｍの吸光度で検出した。
非加熱ＣＮＳＬには、アナカルド酸が６１．８質量％、カルダノールが８．２質量％、カルドールが１９．９質量％、加熱ＣＮＳＬには、アナカルド酸が０．０質量％、カルダノールが７１．４質量％、カルドールが１４．４質量％含まれていた。Production Example A cashew nut shell (500 kg) was obtained from Cashew Trading Co., Ltd. and pressed to produce 158 kg of cashew nut shell oil (non-heated CNSL). Further, heat-treated cashew nut shell liquid (heated CNSL) in which anacardic acid was converted to cardanol by heat treatment at 130 ° C. was obtained from Cashew Trading Co., Ltd. (cold press oil (produced in India)).
The composition of CNSL was measured by the following method. That is, HPLC (Waters 600, Nippon Waters Co., Ltd.), detector (Waters 490E, Nihon Waters Co., Ltd.), printer (Chromatopack C-R6A, Shimadzu Corporation), column (SUPELCOSIL LC18, SUPELCO) were used. A solvent of acetonitrile: water: acetic acid of 80: 20: 1 (volume ratio) was used, and the flow rate was 2 ml / min. Detection was performed at an absorbance of 280 nm.
Non-heated CNSL has 61.8% by weight of anacardic acid, 8.2% by weight of cardanol, 19.9% by weight of cardol, and heated CNSL has 0.0% by weight of anacardic acid and 71.4% of cardanol. % By weight and 14.4% by weight of cardol were contained.
実施例
＜検体の調製＞
リン酸水素二ナトリウム・１２水和物１７．２ｇを脱イオン水に溶解し、５００ｍｌにメスアップして、これを溶液（１）とした。リン酸二水素ナトリウム・２水和物７．５ｇを脱イオン水に溶解し、５００ｍｌにメスアップして、これを溶液（２）とした。溶液（１）に溶液（２）を加えｐＨを７．２に調整して、これを溶液（３）とした。溶液（１）に非加熱ＣＮＳＬ０．２ｇを加え２Ｎ ＨＣｌでｐＨを７．２に調整し、溶液（３）で１００ｍｌにメスアップし、検体として用いた（実施例１）。同様に加熱ＣＮＳＬ０．２ｇを溶液（１）に加え２Ｎ ＨＣｌでｐＨを７．２に調整し、溶液（３）で１００ｍｌにメスアップし、検体として用いた（実施例２）。Example <Sample Preparation>
Disodium hydrogen phosphate dodecahydrate (17.2 g) was dissolved in deionized water, and the volume was made up to 500 ml to obtain a solution (1). 7.5 g of sodium dihydrogen phosphate dihydrate was dissolved in deionized water and made up to 500 ml to obtain a solution (2). Solution (2) was added to solution (1) to adjust the pH to 7.2, which was designated as solution (3). 0.2 g of non-heated CNSL was added to the solution (1), the pH was adjusted to 7.2 with 2N HCl, the solution was made up to 100 ml with the solution (3), and used as a sample (Example 1). Similarly, 0.2 g of heated CNSL was added to the solution (1), the pH was adjusted to 7.2 with 2N HCl, and the solution was made up to 100 ml with the solution (3) and used as a sample (Example 2).
＜インフルエンザウイルスに対する防除効果＞
試験ウイルスにはインフルエンザウイルスＡ型（Ｈ１Ｎ１）ＡＴＣＣ ＶＲ−１６８２を用いた。細胞としてはＭＤＣＫ（ＮＢＬ−２）細胞 ＡＴＣＣ ＣＣＬ−３４株（大日本製薬株式会社）を用いた。
細胞増殖培地にはイーグルＭＥＭ培地「ニッスイ」（日水製薬株式会社）に牛胎仔血清を１０％加えたものを使用した。
細胞維持培地には以下の組成の培地を使用した。<Control effect against influenza virus>
Influenza virus type A (H1N1) ATCC VR-1682 was used as a test virus. MDCK (NBL-2) cell ATCC CCL-34 strain (Dainippon Pharmaceutical Co., Ltd.) was used as the cell.
As the cell growth medium, Eagle's MEM medium “Nissui” (Nissui Pharmaceutical Co., Ltd.) plus 10% fetal calf serum was used.
A medium having the following composition was used as the cell maintenance medium.
イーグルＭＥＭ培地「ニッスイ」 １０００ｍｌ
１０％ＮａＨＣＯ3 １４ｍｌ
Ｌ−グルタミン（３０ｇ／Ｌ） ９．８ｍｌ
１００×ＭＥＭ用ビタミン液 ３０ｍｌ
１０％アルブミン ２０ｍｌ
０．２５％トリプシン ２０ｍｌEagle MEM medium "Nissui" 1000ml
14% 10% NaHCO 3
L-glutamine (30 g / L) 9.8 ml
100 x MEM vitamin solution 30ml
20% 10% albumin
0.25% trypsin 20ml
細胞の培養には細胞増殖培地を用い、使用細胞を組織培養用フラスコ内に単層培養した。単層培養後にフラスコ内から細胞増殖培地を除き、試験ウイルスを接種した。次に細胞維持培地を加えて３７℃の炭酸ガスインキュベーター（ＣＯ2濃度：５％）内で１〜５日間培養した。培養後、倒立位相差顕微鏡を用いて細胞の形態を観察し、細胞に形態変化（細胞変性効果）が起こっていることを確認した。次に、培養液を遠心分離（３０００ｒｐｍ、１０分間）し、得られた上澄み液をウイルス浮遊液とした。
検体１ｍｌにウイルス浮遊液０．１ｍｌを添加・混合し、作用液とした。室温で作用させ、２時間後および６時間後に細胞維持培地を用いて１０００倍に希釈した。なお、脱イオン水を対照として同様に試験し、開始時についても測定を行った。
細胞増殖培地を用い、使用細胞を組織培養用マイクロプレート（９６穴）内で単層培養した後、細胞増殖培地を除き細胞維持培地を０．１ｍｌずつ加えた。次に、１０００倍希釈後の作用液および対照を、細胞維持培地を用いて１０倍段階希釈した。その希釈液０．１ｍｌを４穴ずつに接種し、３７℃の炭酸ガスインキュベーター（ＣＯ2濃度：５％）内で４〜７日間培養した。培養後、倒立位相差顕微鏡を用いて細胞の形態変化（細胞変性効果）の有無を観察し、Ｒｅｅｄ−Ｍｕｅｎｃｈ法により５０％組織培養感染量（ＴＣＩＤ50）を算出して作用液１ｍｌ当たりのウイルス感染価に換算した。
実施例１および実施例２によるインフルエンザウイルス感染価測定結果を表１に示した。Cell growth medium was used for cell culture, and the cells used were cultured in a single layer in a tissue culture flask. After monolayer culture, the cell growth medium was removed from the flask and inoculated with the test virus. Next, a cell maintenance medium was added and cultured in a carbon dioxide incubator (CO 2 concentration: 5%) at 37 ° C. for 1 to 5 days. After culturing, the morphology of the cells was observed using an inverted phase contrast microscope, and it was confirmed that morphological changes (cytopathic effect) occurred in the cells. Next, the culture solution was centrifuged (3000 rpm, 10 minutes), and the resulting supernatant was used as a virus suspension.
0.1 ml of the virus suspension was added to 1 ml of the sample and mixed to obtain a working solution. It was allowed to act at room temperature and diluted 1000-fold with cell maintenance medium after 2 and 6 hours. In addition, it tested similarly using deionized water as a control | contrast, and also measured at the time of a start.
Cell culture medium was used and the cells used were monolayer cultured in a tissue culture microplate (96 wells), and then the cell growth medium was removed and 0.1 ml of cell maintenance medium was added. Next, the working solution and control after 1000-fold dilution were serially diluted 10-fold using a cell maintenance medium. 0.1 ml of the diluted solution was inoculated every 4 holes and cultured in a 37 ° C. carbon dioxide incubator (CO 2 concentration: 5%) for 4 to 7 days. After culturing, the presence or absence of cell morphological change (cytopathic effect) is observed using an inverted phase contrast microscope, the 50% tissue culture infectious dose (TCID 50 ) is calculated by the Reed-Muench method, and the virus per ml of working solution Converted to infectious titer.
The results of measuring the infectious titer of influenza virus according to Example 1 and Example 2 are shown in Table 1.
実施例１および実施例２について、強いインフルエンザウイルス不活性化効果が確認された。これらの結果より、本発明のウイルス感染防除剤が、インフルエンザウイルスの不活化に非常に有効であることが理解される。 About Example 1 and Example 2, the strong influenza virus inactivation effect was confirmed. From these results, it is understood that the virus infection control agent of the present invention is very effective for inactivating influenza virus.
＜ＰＲＲＳウイルスに対する防除効果＞
供試培養細胞にはＭＡ−１０４（アフリカミドリザル腎細胞）を用いた。被検物質には非加熱ＣＮＳＬ ０．２％溶液（実施例１）、および加熱ＣＮＳＬ ０．２％溶液（実施例２）を用いた。ウイルス株にはＰＲＲＳ野外分離株（３型）を用いた。使用培地にはＤＭＥＭ（１０％非働化済み牛胎児血清、１００Ｕ／ｍｌペニシリン、１００Ｕ／ｍｌストレプトマイシン）を用いた。<Control effect against PRRS virus>
MA-104 (African green monkey kidney cells) was used as the test cultured cells. A non-heated CNSL 0.2% solution (Example 1) and a heated CNSL 0.2% solution (Example 2) were used as test substances. A PRRS field isolate (type 3) was used as the virus strain. DMEM (10% inactivated fetal bovine serum, 100 U / ml penicillin, 100 U / ml streptomycin) was used as the medium used.
ウイルス浮遊液の調製は以下の通り行った。細胞の培養はＤＭＥＭ培地で、使用細胞を組織培養用フラスコに単層培養した。単層培養したフラスコに、試験ウイルスを接種した。３７℃の炭酸ガスインキュベーター（ＣＯ2：５％）内で６日間培養した。培養後、培養液を遠心分離（３，０００ｒｐｍ／ｍｉｎ、１０分間）し、得られた上澄み液をウイルス浮遊液とした。The virus suspension was prepared as follows. The cells were cultured in DMEM medium, and the cells used were cultured in a single layer in a tissue culture flask. Monolayer culture flasks were inoculated with the test virus. The cells were cultured for 6 days in a 37 ° C. carbon dioxide incubator (CO 2 : 5%). After the culture, the culture solution was centrifuged (3,000 rpm / min, 10 minutes), and the resulting supernatant was used as a virus suspension.
上記ウイルスを含む上清０．１ｍｌと、前述した非加熱ＣＮＳＬ ０．２％（実施例１）および加熱ＣＮＳＬ ０．２％（実施例２）１ｍＬを混合し、室温で０．５および２時間感作させた。感作液をＤＭＥＭで１０、１００または１，０００倍希釈後、原液または各希釈液をＭＡ−１０４株に感染させた。なお、１試験区につき２連で実施した。無添加区（対照）を同時に設定した。６日間の培養後、培養上清中のＰＲＲＳウイルス濃度をＲｅａｌ−ｔｉｍｅ ＰＣＲで測定した。 0.1 ml of the supernatant containing the virus was mixed with 1 mL of the non-heated CNSL 0.2% (Example 1) and the heated CNSL 0.2% (Example 2) described above, and 0.5 and 2 hours at room temperature. I was sensitized. The sensitizing solution was diluted 10, 100, or 1,000 times with DMEM, and the stock solution or each diluted solution was infected with the MA-104 strain. In addition, it implemented by 2 per test area. An additive-free group (control) was set at the same time. After 6 days of culture, the PRRS virus concentration in the culture supernatant was measured by Real-time PCR.
実施例１および実施例２は、０．５および２時間後のＰＲＲＳウイルスのＬｏｇ ｃｏｐｉｅｓ／ｍｌが１となった。一方、対照区のＬｏｇ ｃｏｐｉｅｓ／ｍｌは０．５時間後で８．４と非常に高い値となった（表２）。この事より、実施例１と実施例２には、高いＰＲＲＳウイルス不活性化効果が確認された。 In Example 1 and Example 2, the log copies / ml of PRRS virus became 1 after 0.5 and 2 hours. On the other hand, Log copies / ml in the control group was very high at 8.4 after 0.5 hours (Table 2). From this, a high PRRS virus inactivating effect was confirmed in Example 1 and Example 2.
供試培養細胞にはＭＡ−１０４（アフリカミドリザル腎細胞）を用いた。被検物質には非加熱ＣＮＳＬ ２５ｐｐｍ溶液（実施例３）を用いた。ウイルス株にはＰＲＲＳ野外分離株（３型）を用いた。使用培地にはＤＭＥＭ（１０％非働化済み牛胎児血清、１００Ｕ／ｍｌペニシリン、１００Ｕ／ｍｌストレプトマイシン）を用いた。
ウイルス浮遊液の調製は以下の通り行った。細胞の培養はＤＭＥＭ培地で、使用細胞を組織培養用フラスコに単層培養した。単層培養したフラスコに、試験ウイルスを接種した。３７℃の炭酸ガスインキュベーター（ＣＯ2：５％）内で６日間培養した。培養後、培養液を遠心分離（３，０００ｒｐｍ／ｍｉｎ、１０分間）し、得られた上澄み液をウイルス浮遊液とした。
上記ウイルスを含む上清１ｍｌと、実施例１及び２と同様にして調製した非加熱ＣＮＳＬ ５０ｐｐｍ溶液１ｍＬを混合して、終濃度２５ｐｐｍとし（実施例３）、室温で２時間感作させた。感作液をＤＭＥＭで１０、１００または１，０００倍希釈後、原液または各希釈液をＭＡ−１０４株に感染させた。なお、１試験区につき３連で実施した。無添加区（対照）を同時に設定した。７日間の培養後、培養上清中のＰＲＲＳウイルス濃度をＲｅａｌ−ｔｉｍｅ ＰＣＲで測定した。MA-104 (African green monkey kidney cells) was used as the test cultured cells. A non-heated CNSL 25 ppm solution (Example 3) was used as a test substance. A PRRS field isolate (type 3) was used as the virus strain. DMEM (10% inactivated fetal bovine serum, 100 U / ml penicillin, 100 U / ml streptomycin) was used as the medium used.
The virus suspension was prepared as follows. The cells were cultured in DMEM medium, and the cells used were cultured in a single layer in a tissue culture flask. Monolayer culture flasks were inoculated with the test virus. The cells were cultured for 6 days in a 37 ° C. carbon dioxide incubator (CO 2 : 5%). After the culture, the culture solution was centrifuged (3,000 rpm / min, 10 minutes), and the resulting supernatant was used as a virus suspension.
1 ml of the supernatant containing the virus and 1 mL of a non-heated CNSL 50 ppm solution prepared in the same manner as in Examples 1 and 2 were mixed to give a final concentration of 25 ppm (Example 3), and sensitized at room temperature for 2 hours. The sensitizing solution was diluted 10, 100, or 1,000 times with DMEM, and the stock solution or each diluted solution was infected with the MA-104 strain. In addition, it implemented in triplicate per test area. An additive-free group (control) was set at the same time. After culturing for 7 days, the PRRS virus concentration in the culture supernatant was measured by Real-time PCR.
実施例３は、２時間後のＰＲＲＳウイルスのＬｏｇ ｃｏｐｉｅｓ／ｍｌが＜１となった。一方、対照区のＬｏｇ ｃｏｐｉｅｓ／ｍｌは２時間後で９．９と非常に高い値となった（表３）。この事より、実施例３には、高いＰＲＲＳウイルス不活性化効果が確認された。 In Example 3, the Log copies / ml of PRRS virus after 2 hours was <1. On the other hand, Log copies / ml in the control group was very high at 9.9 after 2 hours (Table 3). From this, a high PRRS virus inactivating effect was confirmed in Example 3.
本発明のウイルス感染防除剤は、医薬品および飼料の分野で有用である。 The virus infection control agent of the present invention is useful in the fields of pharmaceuticals and feeds.
Claims (8)
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| JP2013543048A JPWO2013069780A1 (en) | 2011-11-11 | 2012-11-09 | Virus infection control agent |
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| JP2013543048A JPWO2013069780A1 (en) | 2011-11-11 | 2012-11-09 | Virus infection control agent |
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Citations (4)
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| JPH02101036A (en) * | 1988-10-05 | 1990-04-12 | Yamanouchi Pharmaceut Co Ltd | Alkylbenzene derivative and medicine therefrom |
| JPH0656657A (en) * | 1992-07-31 | 1994-03-01 | Tonen Corp | Antitumor agent/aids-proofing agent containing fucosidase inhibitor |
| JPH08231410A (en) * | 1995-02-23 | 1996-09-10 | Takasago Internatl Corp | Coccidiosis-reducing agent and feed containing the same |
| WO2011013592A1 (en) * | 2009-07-30 | 2011-02-03 | 出光興産株式会社 | Coated preparation |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU1052492A (en) * | 1991-01-31 | 1992-08-06 | Farmitalia Carlo Erba S.R.L. | Synergistic composition comprising a fibroblast growth factor and a sulfated polylsaccharide, for use as antiviral agent |
| JPH05117150A (en) * | 1991-10-25 | 1993-05-14 | Sanwa Kagaku Kenkyusho Co Ltd | Antiviral agent |
| CN101952310A (en) * | 2007-10-24 | 2011-01-19 | 功能遗传学股份有限公司 | Methods of inhibiting viral infection |
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2012
- 2012-11-09 JP JP2013543048A patent/JPWO2013069780A1/en active Pending
- 2012-11-09 WO PCT/JP2012/079161 patent/WO2013069780A1/en active Application Filing
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| JPH02101036A (en) * | 1988-10-05 | 1990-04-12 | Yamanouchi Pharmaceut Co Ltd | Alkylbenzene derivative and medicine therefrom |
| JPH0656657A (en) * | 1992-07-31 | 1994-03-01 | Tonen Corp | Antitumor agent/aids-proofing agent containing fucosidase inhibitor |
| JPH08231410A (en) * | 1995-02-23 | 1996-09-10 | Takasago Internatl Corp | Coccidiosis-reducing agent and feed containing the same |
| WO2011013592A1 (en) * | 2009-07-30 | 2011-02-03 | 出光興産株式会社 | Coated preparation |
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| ARCH. VIROL., vol. 144, JPN6013003552, 1999, pages 1191 - 1200, ISSN: 0003385558 * |
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