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JP2005119986A - Method for administering physiologically active substance, physiologically active substance-containing emulsion, and method for producing physiologically active substance-containing emulsion - Google Patents

Method for administering physiologically active substance, physiologically active substance-containing emulsion, and method for producing physiologically active substance-containing emulsion Download PDF

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JP2005119986A
JP2005119986A JP2003354482A JP2003354482A JP2005119986A JP 2005119986 A JP2005119986 A JP 2005119986A JP 2003354482 A JP2003354482 A JP 2003354482A JP 2003354482 A JP2003354482 A JP 2003354482A JP 2005119986 A JP2005119986 A JP 2005119986A
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oil
active substance
physiologically active
water
substance
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Teruo Miyazaki
照雄 宮崎
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Japan Science and Technology Agency
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Priority to JP2003354482A priority Critical patent/JP2005119986A/en
Priority to KR1020067007210A priority patent/KR20060123119A/en
Priority to CNA2004800301817A priority patent/CN1867319A/en
Priority to PCT/KR2004/002652 priority patent/WO2005034914A1/en
Publication of JP2005119986A publication Critical patent/JP2005119986A/en
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/10Dispersions; Emulsions
    • A61K9/107Emulsions ; Emulsion preconcentrates; Micelles

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Dispersion Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
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  • Veterinary Medicine (AREA)
  • Medicinal Preparation (AREA)
  • Feed For Specific Animals (AREA)
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Abstract

<P>PROBLEM TO BE SOLVED: To provide an excellent oral drug delivery technique utilizing an oil particle-absorbing action in a constant animal digestive canal. <P>SOLUTION: This method for administering the physiologically active substance is characterized by preparing an oil-in-water type emulsion whose oil drops are fine oil particles containing the physiologically active substance and having an average particle diameters targeted for an oil sphere-absorbing action in the digestive canals of animals, and then orally administering the emulsion to a non-human animal whose oil particle-absorbing action has been recognized. In the fine oil particles in the emulsion, the physiologically active substance is buried in the oil phase by the action of an oil-soluble emulsifier as such or while forming an extremely fine dispersion phase together with a small amount of water. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

本願発明は、本願発明者の報告によって比較的最近に知られるに到った油球吸収作用を利用する、動物への経口的ドラッグデリバリー技術に関する。より詳しくは、本願発明は、一定の動物の消化管における油球吸収作用を有効に利用した生理活性物質の投与方法、このような投与方法に特に良好に適合すべく新規に開発された生理活性物質含有乳化液、及びこの生理活性物質含有乳化液の製造方法に関する。   The present invention relates to a technique for oral drug delivery to animals utilizing the oil-ball absorbing action which has been known relatively recently by the report of the present inventor. More specifically, the present invention relates to a method for administering a physiologically active substance that effectively utilizes the action of oil ball absorption in the digestive tract of certain animals, and a newly developed physiological activity that is particularly well suited to such a method of administration. The present invention relates to a substance-containing emulsion and a method for producing this physiologically active substance-containing emulsion.

昔から、例えばヒト、ウサギ、マウス、家畜等の哺乳動物の特定の疾病を予防するため、その疾病に対するワクチンや免疫賦活物質等を投与する方法が種々に提案されている。近年は、ワクチンや一定の薬理作用を示す物質その他各種の生理活性物質を養殖水棲動物等に投与して、その疾病の予防・治療等を図ろうとする試みも多くなっている。   For a long time, various methods for administering vaccines, immunostimulatory substances, and the like have been proposed to prevent specific diseases of mammals such as humans, rabbits, mice, and domestic animals. In recent years, many attempts have been made to administer vaccines, substances exhibiting a certain pharmacological action, and various other physiologically active substances to cultured aquatic animals and the like to prevent or treat the disease.

ヒトその他の哺乳動物に対して伝統的に行われている血管注射(静脈注射)等の方法は、理論的には優れた効果を期待できるドラッグデリバリー方法であるが、生物体に対して一定の外科的侵襲ストレスを与えると言う欠点があり、更に、この方法の魚類や甲殻類、貝類等への適用は容易ではない。   Methods such as vascular injection (intravenous injection) traditionally performed on humans and other mammals are drug delivery methods that can theoretically be expected to have excellent effects. There is a drawback of applying surgically invasive stress, and furthermore, this method is not easily applied to fish, shellfish, shellfish, and the like.

従って、特に魚類、甲殻類、貝類等に対する生理活性物質の投与方法としては、経口投与が主流となっている。そして、その経口投与剤を設計するに当たっては、投与すべき1種類又は2種類以上の生理活性物質や、これと併用する種々の助剤がそれぞれに異なる親和性(親水性あるいは親油性)を持つ点等を考慮して、経口投与剤の安定性等の配慮から、各種の形態のエマルジョン製剤を工夫している場合が多い。   Therefore, oral administration has become the mainstream as a method for administering physiologically active substances to fish, shellfish, shellfish and the like. And in designing the oral administration agent, one or two or more types of physiologically active substances to be administered and various auxiliary agents used in combination with each have different affinity (hydrophilicity or lipophilicity). Considering the points, etc., various forms of emulsion preparations are often devised in consideration of the stability of orally administered agents.

特開平11−255664号公報 上記の特許文献1には、分子量が10,000以下のペプチドグリカンを経口投与用免疫増強剤の有効成分として、これをそのまま、あるいは飼料に添加して、哺乳動物や魚類、甲殻類(例えば、エビの一種であるブラックタイガー)に投与する技術が開示されている。In JP-A-11-255664, peptidoglycan having a molecular weight of 10,000 or less is used as an active ingredient of an immunopotentiator for oral administration, and it is added as it is to a feed, to mammals and fish. A technique for administration to crustaceans (for example, black tiger, a kind of shrimp) is disclosed.

特表平8−504811号公報 上記の特許文献2には、生理的に許容できる薬剤送出用エマルジョンにして、フルオロケミカル(例えば、パーフルオロデカリン等のパーフルオロカーボン)及び油の水中に乳化された粒子、界面活性剤並びに該エマルジョンに可溶化されている薬剤を含んでなるエマルジョンが開示されている。In JP-A-8-504811, the above-mentioned patent document 2 describes a particle that is made into a physiologically acceptable emulsion for drug delivery and emulsified in water of fluorochemical (for example, perfluorocarbon such as perfluorodecalin) and oil. , An emulsion comprising a surfactant and a drug solubilized in the emulsion is disclosed.

この特許文献2に係る発明は、曖昧かつ難解な文章表現からなるが、要するに、トリグリセリドのような液状脂肪油30重量%以下をマトリクスとし、これに対して75重量%以下のフルオロケミカル液と、30重量%以下の薬剤液とが界面活性剤の作用により分散しているエマルジョン系が記載されている。   The invention according to this Patent Document 2 consists of ambiguous and esoteric text expressions, but in short, a liquid fatty oil such as triglyceride 30% by weight or less as a matrix, and 75% by weight or less of a fluorochemical liquid, An emulsion system in which 30% by weight or less of a drug solution is dispersed by the action of a surfactant is described.

特開平7−53404号公報 上記の特許文献3は、抗原性物質及び一方のアジュバントが油型であり他方のアジュバントが水型である油型及び水型アジュバント混合物の発明を開示する。より具体的には、油中水滴型エマルジョンであって、マトリクスたる分散媒(例えば70重量%の油)には乳化剤を含ませることにより、分散質(例えば30重量%の水滴)には抗原性物質及び/又はアジュバントを含ませている。JP-A-7-53404 discloses the invention of an oil-type and water-type adjuvant mixture in which an antigenic substance and one adjuvant are oil-type and the other adjuvant is water-type. More specifically, the emulsion is a water-in-oil emulsion, and the dispersoid (eg, 30% by weight of water droplets) is made antigenic by adding an emulsifier to the matrix dispersion medium (eg, 70% by weight of oil). Substances and / or adjuvants are included.

しかし、生理活性物質を経口投与した場合、その投与の効果が十分には発現し難いと言う一般的な問題が以前から認識されている。例えば上記の特許文献1の一部にも、魚類に対する生理活性物質の経口投与が十分な効果を生じ難い旨の指摘が見られる。その理由は必ずしも完全に解明されていないが、恐らく、血管注射とは異なり、経口投与の場合は消化管を通じての吸収となるため、生理活性物質が吸収の前に消化されたり、消化管で十分には吸収されなかったりする点に関連しているものと思われる。   However, when a physiologically active substance is orally administered, a general problem that the effect of the administration is not sufficiently expressed has been recognized for some time. For example, it is pointed out that a part of the above-mentioned Patent Document 1 is difficult to produce a sufficient effect by oral administration of a physiologically active substance to fish. The reason for this is not always fully understood, but perhaps unlike vascular injection, oral administration results in absorption through the gastrointestinal tract, so that bioactive substances are digested before absorption or sufficient in the gastrointestinal tract. It seems to be related to the point that it is not absorbed.

ところで、一般的に、動物の消化管においては、油(脂肪)は脂肪酸とグリセリンに分解された後に分子レベルで吸収される、とするのが旧来の常識である。しかしながら、本願発明者は、魚類等の消化管において、一定の微小なサイズの油球が腸管上皮からそのまま体内に取り込まれると言う、驚くべき油球吸収作用を見出した。この知見は、下記の非特許文献1等において報告している。その後、一部の哺乳動物においても同様の油球吸収作用が認められるとする新規な追加的知見も得られ始めている。   By the way, in general, in the digestive tract of animals, it is a conventional common sense that oil (fat) is absorbed at the molecular level after being decomposed into fatty acid and glycerin. However, the inventor of the present application has found a surprising oil ball absorption action that a certain minute size of oil balls is taken directly into the body from the intestinal epithelium in the digestive tract of fish and the like. This finding is reported in the following Non-Patent Document 1 and the like. Since then, new additional findings have begun to be obtained that a similar oil-ball absorbing action is observed in some mammals.

「マダイおよびクロダイ仔稚魚の卵黄吸収と消化管での栄養摂取に関する組織学的研究」 宮崎照雄ら 三重大学生物資源学部紀要1号、15−27頁(1988) 本願発明者は、このような油球吸収作用を巧妙に利用すれば、ワクチン、免疫賦活物質や薬理活性物質等の生理活性物質を、簡易な手段により非常に高い効率で動物に摂取させる経口的ドラッグデリバリー手段を構築できるのではないか、との着想に到り、本願発明を完成した。“Histological study on egg yolk absorption and nutrient intake in digestive tract of red sea bream and black sea bream larvae” Teruo Miyazaki et al. Bulletin of Faculty of Bioresources, Mie University, pp. 15-27 (1988) If skillful use of the sphere absorption action, it is not possible to construct an oral drug delivery method that allows animals to ingest physiologically active substances such as vaccines, immunostimulatory substances and pharmacologically active substances with a very high efficiency. The present invention has been completed.

なお、このような着想においては、(イ)その適用対象を消化管での油球吸収作用が確認されている動物に限ること、(ロ)対象動物に経口投与する微小油球が、油球吸収作用の対象となる一定の平均粒径を有すること、の2点が絶対的な条件である。上記の特許文献1〜特許文献3等の従来技術においては、この2点の条件は全く開示されていないし、考慮されてもいない。又、本願発明者の報告に係る上記の非特許文献1は、油球吸収作用を報告するが、この作用を経口的なドラッグデリバリー手段に利用できる可能性には全く触れていないし、もちろん、そのための具体的な実施手段も全く検討していない。   In such an idea, (a) the application target is limited to animals that have been confirmed to absorb oil balls in the gastrointestinal tract, and (b) micro oil balls orally administered to the target animals are oil balls. There are two absolute conditions: having a certain average particle diameter to be the object of absorption. In the conventional techniques such as Patent Document 1 to Patent Document 3 described above, these two conditions are not disclosed at all and are not taken into consideration. Further, the above-mentioned Non-Patent Document 1 relating to the report of the present inventor reports the oil ball absorption action, but does not mention the possibility that this action can be used for oral drug delivery means. The specific implementation method is not considered at all.

(第1発明の構成)
上記課題を解決するための本願第1発明の構成は、水中油滴型の乳化液であって、その油滴が生理活性物質を含むと共に動物の消化管における油球吸収作用の対象となる平均粒径を有する微小油球であるものを調製し、この乳化液を、消化管における油球吸収作用が認められている非ヒト動物に経口投与する、生理活性物質の投与方法である。
(Configuration of the first invention)
The configuration of the first invention of the present application for solving the above-mentioned problem is an oil-in-water type emulsion, and the oil droplet contains a physiologically active substance and is an average subject to oil ball absorption action in the digestive tract of animals. This is a method for administering a physiologically active substance, in which a fine oil sphere having a particle size is prepared and this emulsion is orally administered to a non-human animal in which an action of absorbing oil globules in the digestive tract is recognized.

(第2発明の構成)
上記課題を解決するための本願第2発明の構成は、前記第1発明に係る微小油球が、1〜60μmの範囲内の平均粒径を有する、生理活性物質の投与方法である。
(Configuration of the second invention)
The configuration of the second invention of the present application for solving the above problem is a method for administering a physiologically active substance, in which the micro oil spheres according to the first invention have an average particle diameter in the range of 1 to 60 μm.

(第3発明の構成)
上記課題を解決するための本願第3発明の構成は、前記第1発明又は第2発明に係る微小油球が以下(1)〜(3)のいずれかの粒子構造を持つものである、生理活性物質の投与方法である。
(1)微小油球中において、油溶性乳化剤の作用により、生理活性物質がそのまま、あるいは少量の水分と共に、極めて微小な分散相を形成している。
(2)微小油球中において、親油性の生理活性物質が溶解又は懸濁している。
(3)微小油球が脂質二重層によって形成されたミセル構造を持ち、その内部に生理活性物質が封入されている。
(Configuration of the third invention)
The configuration of the third invention of the present application for solving the above-described problem is that the micro oil sphere according to the first invention or the second invention has a particle structure of any one of (1) to (3) below. This is a method for administering an active substance.
(1) In the fine oil sphere, the bioactive substance forms an extremely fine dispersed phase as it is or with a small amount of water by the action of the oil-soluble emulsifier.
(2) A lipophilic physiologically active substance is dissolved or suspended in the fine oil sphere.
(3) A micro oil sphere has a micelle structure formed by a lipid bilayer, and a physiologically active substance is enclosed therein.

上記の(1)において、「油溶性乳化剤の作用により」とは、微小油球の内部において、生理活性物質の(又は、生理活性物質と少量の水分との)極めて微小な分散相が、その個々の分散相における分散質との界面に油溶性乳化剤を保持した状態で、微小油球の分散質たる油相の中に安定的に分散している、と言う油溶性乳化剤の作用を言う。   In the above (1), “by the action of an oil-soluble emulsifier” means that an extremely minute dispersed phase of a physiologically active substance (or a physiologically active substance and a small amount of water) is contained in the inside of a fine oil sphere. The action of the oil-soluble emulsifier means that the oil-soluble emulsifier is stably dispersed in the oil phase as the dispersoid of the fine oil spheres while the oil-soluble emulsifier is held at the interface with the dispersoid in each dispersed phase.

(第4発明の構成)
上記課題を解決するための本願第4発明の構成は、前記第1発明〜第3発明のいずれかに係る乳化液を、飼料又は飼料用ペレットに含浸させたもとで動物に摂食させる、生理活性物質の投与方法である。
(Configuration of the fourth invention)
The structure of the fourth invention of the present application for solving the above-described problem is that the animal feeds a feed or feed pellet impregnated with the emulsion according to any of the first to third inventions. It is a method of administration of a substance.

(第5発明の構成)
上記課題を解決するための本願第5発明の構成は、前記第1発明〜第4発明のいずれかに係る動物が、水棲動物又は非ヒト陸棲哺乳動物である、生理活性物質の投与方法である。
(Structure of the fifth invention)
The configuration of the fifth invention of the present application for solving the above-mentioned problem is a method for administering a physiologically active substance, wherein the animal according to any one of the first to fourth inventions is an aquatic animal or a non-human terrestrial mammal. .

(第6発明の構成)
上記課題を解決するための本願第6発明の構成は、前記第1発明〜第5発明のいずれかに係る生理活性物質が、ワクチン、免疫賦活物質、栄養物質、薬理作用物質、天然色素又はミネラルである、生理活性物質の投与方法である。
(Structure of the sixth invention)
The structure of the sixth invention of the present application for solving the above problem is that the physiologically active substance according to any one of the first to fifth inventions is a vaccine, an immunostimulatory substance, a nutritional substance, a pharmacologically active substance, a natural pigment or a mineral. This is a method for administering a physiologically active substance.

(第7発明の構成)
上記課題を解決するための本願第7発明の構成は、動物の消化管における油球吸収作用の対象となる平均粒径を有する微小油球が分散した水中油滴型の乳化液であって、該微小油球中においては、油溶性乳化剤の作用により、生理活性物質がそのまま、あるいは少量の水分と共に極めて微小な分散相を形成して油相中に包埋されている、生理活性物質含有乳化液である。ここに、「油溶性乳化剤の作用により」とは、「第3発明の構成」欄において説明した通りの意味である。
(Structure of the seventh invention)
The structure of the seventh invention of the present application for solving the above-mentioned problem is an oil-in-water emulsion in which fine oil spheres having an average particle size to be subjected to oil sphere absorption action in an animal digestive tract are dispersed, In the micro oil sphere, the bioactive substance-containing emulsification is embedded in the oil phase as it is or by forming an extremely fine dispersed phase with a small amount of water by the action of the oil-soluble emulsifier. It is a liquid. Here, “by the action of the oil-soluble emulsifier” has the same meaning as described in the “Structure of the third invention” column.

(第8発明の構成)
上記課題を解決するための本願第8発明の構成は、前記第7発明に係る微小油球が、1〜60μmの範囲内の平均粒径を有する、生理活性物質含有乳化液である。
(Configuration of the eighth invention)
The configuration of the eighth invention of the present application for solving the above problem is a bioactive substance-containing emulsion in which the fine oil spheres according to the seventh invention have an average particle diameter in the range of 1 to 60 μm.

(第9発明の構成)
上記課題を解決するための本願第9発明の構成は、前記第7発明又は第8発明に係る生理活性物質が、ワクチン、免疫賦活物質、栄養物質、薬理作用物質、天然色素又はミネラルである、生理活性物質含有乳化液である。
(Structure of the ninth invention)
The structure of the ninth invention of the present application for solving the above problems is that the physiologically active substance according to the seventh or eighth invention is a vaccine, an immunostimulatory substance, a nutritional substance, a pharmacologically active substance, a natural pigment or a mineral, It is a bioactive substance-containing emulsion.

(第10発明の構成)
上記課題を解決するための本願第10発明の構成は、以下の(A)第1乳化工程と、(B)第2乳化工程とを含む、生理活性物質含有乳化液の製造方法である。
(A)第1乳化工程:生理活性物質又はその水溶液もしくは水懸濁液と、油溶性乳化剤を添加した油液とを混合して乳化させることにより、生理活性物質がそのまま、あるいは少量の水分と共に極めて微小な分散相を形成して油相中に包埋されている油性乳化相を製造する。
(B)第2乳化工程:第1乳化工程で製造された油性乳化相と、水溶性乳化剤を添加した水とを混合して乳化させることにより、前記油性乳化相が動物の消化管における油球吸収作用の対象となる平均粒径を持つ微小油球として水中に分散した水中油滴型の乳化液を製造する。
(Configuration of the tenth invention)
The configuration of the tenth invention of the present application for solving the above problems is a method for producing a physiologically active substance-containing emulsion comprising the following (A) first emulsification step and (B) second emulsification step.
(A) 1st emulsification process: By mixing and emulsifying the bioactive substance or its aqueous solution or water suspension and the oil liquid to which the oil-soluble emulsifier is added, the bioactive substance remains as it is or with a small amount of water. An oily emulsified phase is produced which forms a very fine dispersed phase and is embedded in the oil phase.
(B) Second emulsification step: The oily emulsified phase produced in the first emulsification step and water added with a water-soluble emulsifier are mixed and emulsified, whereby the oily emulsified phase becomes an oil ball in the digestive tract of animals. An oil-in-water emulsion that is dispersed in water as fine oil spheres having an average particle size to be absorbed is produced.

ここにおいて、上記(A)における油相中においては、及び上記(B)における微小油球中においては、生理活性物質の(又は、生理活性物質と少量の水分との)極めて微小な分散相が、その個々の分散相における分散質との界面に油溶性乳化剤を保持した状態で、分散質たる油相又は微小油球の中に安定的に分散している。又、上記(B)における水中においては、このような微小油球が、その個々の微小油球における分散質(水)との界面に水溶性乳化剤を保持した状態で、分散質たる水中に安定的に分散している。   Here, in the oil phase in the above (A) and in the micro oil sphere in the above (B), an extremely minute dispersed phase of the physiologically active substance (or the physiologically active substance and a small amount of water) is present. In each of the dispersed phases, the oil-soluble emulsifier is held at the interface with the dispersoid, and the oil is stably dispersed in the oil phase or the fine oil spheres as the dispersoid. Further, in the water in the above (B), such fine oil spheres are stable in the water as the dispersoid while holding the water-soluble emulsifier at the interface with the dispersoids (water) in the individual fine oil spheres. Distributed.

(第1発明の効果)
第1発明の生理活性物質の投与方法は、経口投与法であるから、投与対象である非ヒト動物に対して血管注射法のような外科的侵襲ストレスを与えないし、例えば魚類や甲殻類、貝類等への適用も容易である。
(Effect of the first invention)
Since the administration method of the physiologically active substance of the first invention is an oral administration method, it does not give a surgical invasive stress like a vascular injection method to a non-human animal to be administered, for example, fish, shellfish, shellfish Etc. are also easy to apply.

次に、第1発明の最も重要な特徴として、この方法は、動物の消化管における油球吸収作用を有効に利用する経口投与法である。即ち、第1発明においては、前記の(イ)その適用対象を消化管での油球吸収作用が確認されている動物に限ること、(ロ)対象動物に経口投与する微小油球が、油球吸収作用の対象となる一定の平均粒径を有すること、の2点の条件が満たされ、かつ、ドラッグデリバリーの目的物質たる生理活性物質が水中油滴型乳化液の油滴たる微小油球中に含まれている。   Next, as the most important feature of the first invention, this method is an oral administration method that effectively utilizes the oil-ball absorbing action in the digestive tract of animals. That is, in the first invention, (b) the application target is limited to animals that have been confirmed to absorb oil balls in the digestive tract, and (b) micro oil balls that are orally administered to the target animals Micro oil spheres satisfying the two conditions of having a certain average particle size to be a target of sphere absorption, and the physiologically active substance as a drug delivery target substance being oil droplets of an oil-in-water emulsion Included in.

このため、経口投与した対象動物の消化管において、生理活性物質を含む微小油球は丸ごと腸管上皮からそのまま体内に取り込まれる。その結果、従来の経口投与方法に見られた、生理活性物質が吸収の前に消化されたり、消化管で十分には吸収されなかったりすると言う不具合を回避することができ、生理活性物質を簡易な手段により非常に高い効率で動物体内に摂取させることができる。   For this reason, in the digestive tract of an orally administered subject animal, the entire micro oil sphere containing a physiologically active substance is directly taken into the body from the intestinal epithelium. As a result, it is possible to avoid the disadvantage that the physiologically active substance is digested before absorption or is not sufficiently absorbed in the digestive tract, which is seen in the conventional oral administration method. Can be taken into the animal body with extremely high efficiency.

(第2発明の効果)
上記の第1発明における「動物の消化管における油球吸収作用の対象となる平均粒径」は、実際には動物種によって相違があるため、数値を以て一律に規定することが困難である。しかしながら、あえて数値を以て規定すれば、一般的には、1〜60μmの範囲内の平均粒径を有することが好ましい。
(Effect of the second invention)
Since the “average particle size that is the target of oil ball absorption in the digestive tract of animals” in the first invention is actually different depending on the animal species, it is difficult to uniformly define the numerical values. However, it is generally preferable to have an average particle size in the range of 1 to 60 μm if it is defined with numerical values.

(第3発明の効果)
油球吸収作用の対象となる「油球」の種類あるいは粒子構造は限定されないが、例えば第3発明の(1)〜(3)に列挙したものが、いずれも油球分散液としての安定性を期待できることから、代表的に例示される。
(Effect of the third invention)
There are no limitations on the type or particle structure of the “oil ball” that is the target of the oil ball absorption action. For example, those listed in (1) to (3) of the third invention are all stable as an oil ball dispersion. Is representatively exemplified.

但し、これらの(1)〜(3)の粒子構造を相対的に評価したとき、(3)は一般的に「リポソーム」と通称される小胞状の粒子構造であるが、動物の腸管内リパーゼの作用で境界膜(脂質二重層)が崩壊した場合には油球構造を維持できず、油球吸収作用を利用できなくなると言う欠点がある。(2)の粒子構造は、要するに油のボールであるから、リパーゼの作用によってもその油球構造は容易に崩壊しないが、油溶性の生理活性物質を油球中に溶解させる場合は別として、一般的には生理活性物質を油球中に微分散させることが困難であり、ひいては、所定の微小な平均粒径を有する油球を調製すること自体も困難になり易いと言う欠点がある。   However, when these particle structures (1) to (3) are relatively evaluated, (3) is a vesicle-like particle structure generally called “liposome”, but it is an animal intestinal lipase. When the boundary membrane (lipid bilayer) is collapsed by the above action, the oil ball structure cannot be maintained, and the oil ball absorption function cannot be used. Since the particle structure of (2) is essentially an oil ball, the oil sphere structure is not easily collapsed by the action of lipase, except when an oil-soluble physiologically active substance is dissolved in the oil sphere, In general, it is difficult to finely disperse a physiologically active substance in an oil sphere, and as a result, it is difficult to prepare an oil sphere having a predetermined minute average particle diameter.

これらに比較して、(1)の粒子構造は本願発明者が第1発明の効率的実施のために創製した粒子構造であって、その詳細は第7発明において述べるが、動物の腸管内リパーゼの作用によっても油球構造は崩壊せず、しかも生理活性物質の親和性の如何を問題とせずに、これを微小油球中に極めて微小に分散させることができると言うメリットがある。   Compared with these, the particle structure of (1) is a particle structure created by the present inventor for the efficient implementation of the first invention, and the details thereof will be described in the seventh invention. The effect of this is that the structure of the oil sphere does not collapse, and there is an advantage that it can be dispersed very finely in the micro oil sphere without considering the affinity of the physiologically active substance.

(第4発明の効果)
第1発明〜第3発明に係る乳化液の経口投与方法は限定されず、例えば対象動物たる魚類等の口腔に注射器等で注入すると言う方法も可能であるが、第4発明のように、乳化液を飼料又は飼料用ペレットに含浸させたもとで動物に摂食させる方法が、特に好ましい。この場合の飼料用ペレットの材質は限定されないが、多孔質のペレットが特に好ましい。
(Effect of the fourth invention)
The method for oral administration of the emulsion according to the first to third inventions is not limited, and for example, a method of injecting into the oral cavity of fish or the like as the target animal with a syringe or the like is possible, but as in the fourth invention, emulsification Particularly preferred is a method in which an animal is fed with the liquid impregnated in feed or feed pellets. The material of the pellet for feed in this case is not limited, but a porous pellet is particularly preferable.

(第5発明の効果)
第1発明〜第4発明における適用対象たる動物の種類は、消化管における油球吸収作用が認められている非ヒト動物である限りにおいて限定されないが、水棲動物又は非ヒト陸棲哺乳動物が、特に好ましい。
(Effect of the fifth invention)
The type of animal to be applied in the first invention to the fourth invention is not limited as long as it is a non-human animal in which the action of absorbing oil balls in the digestive tract is recognized, but aquatic animals or non-human terrestrial mammals are particularly preferred. preferable.

(第6発明の効果)
第1発明〜第5発明で利用する生理活性物質の種類は限定されないが、ワクチン、免疫賦活物質、栄養物質、薬理作用物質、天然色素又はミネラル等を好ましく例示することができる。
(Effect of the sixth invention)
Although the kind of physiologically active substance utilized by 1st invention-5th invention is not limited, A vaccine, an immunostimulatory substance, a nutrient substance, a pharmacological action substance, a natural pigment | dye, a mineral, etc. can be illustrated preferably.

(第7発明の効果)
第7発明の生理活性物質含有乳化液は、水中油滴型の乳化液であるため、油滴たる微小油球の分散状態が安定している。又、その微小油球は、動物の消化管における油球吸収作用の対象となる平均粒径を有するため、対象動物の消化管において良好に油球吸収作用を利用でき、高い効率で対象動物の体内に摂取されることが可能である。
(Effect of the seventh invention)
Since the bioactive substance-containing emulsion of the seventh invention is an oil-in-water emulsion, the dispersion state of the fine oil spheres as oil droplets is stable. In addition, since the micro oil spheres have an average particle size that is the target of the oil ball absorption action in the digestive tract of animals, the oil ball absorption action can be used well in the digestive tract of the target animal, and the target animal can be efficiently used. It can be taken into the body.

しかも、この微小油球中においては、油溶性乳化剤の作用により、生理活性物質がそのまま、あるいは少量の水分と共に極めて微小な分散相を形成して油相中に包埋されている。従って、第1に、動物の腸管内リパーゼの作用を受けた場合、微小油球の表層部の一部が消化されたとしても、微小油球の粒子構造はミセル構造のようには崩壊せずに維持され、腸管上皮より良好に取込まれる。第2に、生理活性物質を油溶性乳化剤の作用により油中に分散させるので、生理活性物質を、その親和性(親水性又は親油性)の如何に係わらず、しかも極めて微細な分散状態で、微小油球中に包埋させることができ、ひいては所定の微小な平均粒径を有する油球を調製することも容易となる。   In addition, in this micro oil sphere, the physiologically active substance is embedded in the oil phase as it is or by forming a very fine dispersed phase with a small amount of water by the action of the oil-soluble emulsifier. Therefore, first, when subjected to the action of lipase in the intestinal tract of an animal, even if a part of the surface layer of the microsphere is digested, the particle structure of the microsphere does not collapse like a micelle structure. And is better taken up than the intestinal epithelium. Second, since the physiologically active substance is dispersed in the oil by the action of the oil-soluble emulsifier, the physiologically active substance can be dispersed in an extremely fine dispersion state regardless of its affinity (hydrophilic or lipophilic). It can be embedded in the fine oil spheres, and as a result, it becomes easy to prepare oil globules having a predetermined fine average particle diameter.

(第8発明の効果)
上記の第7発明における「動物の消化管における油球吸収作用の対象となる平均粒径」は、実際には動物種によって相違があるため、数値を以て一律に規定することが困難である。しかしながら、あえて数値を以て規定すれば、一般的には、1〜60μmの範囲内の平均粒径を有することが好ましい。
(Effect of the eighth invention)
Since the “average particle diameter that is the target of the oil-ball absorbing action in the digestive tract of animals” in the seventh invention is actually different depending on the animal species, it is difficult to uniformly define the numerical values. However, it is generally preferable to have an average particle size in the range of 1 to 60 μm if it is defined with numerical values.

(第9発明の効果)
第7発明又は第8発明で利用する生理活性物質の種類は限定されないが、ワクチン、免疫賦活物質、栄養物質、薬理作用物質、天然色素又はミネラルを好ましく例示することができる。
(Effect of the ninth invention)
Although the kind of the physiologically active substance used in the seventh invention or the eighth invention is not limited, a vaccine, an immunostimulatory substance, a nutrient substance, a pharmacological agent, a natural pigment or a mineral can be preferably exemplified.

(第10発明の効果)
第10発明の製造方法によって、第7発明〜第9発明に係る生理活性物質含有乳化液を有効に製造することができる。
(Effect of the tenth invention)
The bioactive substance-containing emulsion according to the seventh to ninth inventions can be effectively produced by the production method of the tenth invention.

第10発明において、第1乳化工程における技術的なキーポイントは、第1に、生理活性物質又はその水溶液等と、油液とを混合することである。第2に、その油液には油溶性乳化剤を添加しておくことである。この2点のキーポイントを具備したとき、超音波処理やホモジナイザー利用等の公知の適宜な乳化液形成手段の利用により、「生理活性物質がそのまま、あるいは少量の水分と共に極めて微小な分散相を形成して油相中に包埋されている油性乳化相」を構成することができる。   In the tenth aspect of the invention, the technical key point in the first emulsification step is to first mix a bioactive substance or an aqueous solution thereof with an oil solution. Second, an oil-soluble emulsifier is added to the oil solution. When these two key points are provided, by using a known appropriate emulsion forming means such as ultrasonic treatment or using a homogenizer, “a physiologically active substance is formed as it is or with a small amount of water to form a very fine dispersed phase. Thus, an “oil-based emulsified phase embedded in the oil phase” can be constituted.

又、第2乳化工程においては、上記の油性乳化相と、水(水溶性乳化剤を添加しておく)とを混合し、公知の適宜な乳化液形成手段の利用により乳化させることにより、第7発明〜第9発明に係る生理活性物質含有乳化液を製造することができる。   In the second emulsification step, the above-mentioned oily emulsified phase and water (with a water-soluble emulsifier added) are mixed and emulsified by using a known appropriate emulsion forming means. The bioactive substance-containing emulsion according to the invention to the ninth invention can be produced.

次に、本願の第1発明〜第10発明を実施するための形態を、その最良の形態を含めて説明する。以下において、単に「本発明」と言う時は、本願の各発明を一括して指している。   Next, modes for carrying out the first invention to the tenth invention of the present application will be described including the best mode. In the following, the term “present invention” refers to each invention of the present application collectively.

〔生理活性物質の投与方法〕
本発明に係る生理活性物質の投与方法においては、水中油滴型の乳化液であってその油滴が生理活性物質を含むと共に動物の消化管における油球吸収作用の対象となる平均粒径を有する微小油球であるものを調製し、この乳化液を、消化管における油球吸収作用が認められている非ヒト動物に経口投与する。
[Method of administering physiologically active substance]
In the method for administering a physiologically active substance according to the present invention, an oil-in-water type emulsion, wherein the oil droplets contain a physiologically active substance and have an average particle size that is a target for oil ball absorption action in the digestive tract of animals. A fine oil sphere is prepared, and this emulsion is orally administered to a non-human animal in which the action of absorbing oil globules in the digestive tract is recognized.

なお、本発明において用いる各種の材料、例えば、乳化液の油滴を構成する油液、乳化液の形成に用いる界面活性剤(後述する生理活性物質含有乳化液の形成に用いる油溶性乳化剤及び水溶性乳化剤を含む)、あるいは飼料用ペレット等は、いずれも対象動物に対する無毒性又は生理的安全性が確認されたものを用いることを前提としている。更に、栄養面等で対象動物に有利なものを用いることが、より望ましい。   In addition, various materials used in the present invention, for example, an oil liquid constituting an oil droplet of an emulsion liquid, a surfactant used for forming an emulsion liquid (an oil-soluble emulsifier and a water-soluble emulsifier used for forming a bioactive substance-containing emulsion liquid described later) It is premised that feed pellets and the like are used that have been confirmed to be non-toxic or physiologically safe for target animals. Furthermore, it is more desirable to use a thing advantageous to the target animal in terms of nutrition and the like.

本発明に係る生理活性物質の投与方法において、水中油滴型乳化液の経口投与の具体的な方法は限定されない。例えば、注射器等を用いて対象動物の口腔内に注入することもできるし、哺乳動物に対しては液剤のまま飲用させることも可能である。又、その動物用の飼料に混ぜて摂食させることもできる。より好ましい方法は、その動物用の飼料ペレット(特に好ましくは、多孔質の飼料ペレット)に含浸させて、そのまま、あるいは乾燥させてから摂食させる方法である。飼料ペレットとしては、各種の市販品等を任意に利用できる。   In the administration method of the physiologically active substance according to the present invention, the specific method of oral administration of the oil-in-water emulsion is not limited. For example, it can be injected into the oral cavity of the target animal using a syringe or the like, and can be drunk as a liquid for mammals. It can also be mixed with the animal feed. A more preferred method is a method of impregnating the animal feed pellets (particularly preferably, porous feed pellets) and feeding them as they are or after drying. As the feed pellet, various commercially available products can be arbitrarily used.

〔生理活性物質〕
生理活性物質の種類は限定される必然性がなく、全く任意に選択することができる。上位概念的なカテゴリーとしては、ワクチン、免疫賦活物質、栄養物質、薬理作用物質、天然色素又はミネラル等を好ましく例示することができる。
[Bioactive substances]
The type of the physiologically active substance is not necessarily limited, and can be selected arbitrarily. Preferred examples of the superordinate category include vaccines, immunostimulatory substances, nutritional substances, pharmacologically active substances, natural pigments or minerals.

より具体的には、例えばワクチンとしての病原ウイルスや細菌又は寄生虫の粉砕物等、グルカン、フィコイダン、LPS(リポ多糖類)等の免疫賦活剤を例示できる。抗体等の生理活性を有するポリペプチドも例示できる。ビタミン類等の栄養物質も例示できる。抗生物質、ホルモン、インターフェロン等の薬理作用物質も例示できる。その他にも、対象動物に一定の色彩を賦与するための天然色素等も例示できる。有機態又は無機態のカルシウム、鉄、マグネシウム等のミネラルも例示できる。   More specifically, for example, immunostimulators such as glucan, phycoidan, LPS (lipopolysaccharide), etc., such as pathogenic virus as a vaccine, pulverized product of bacteria or parasite, and the like can be exemplified. Polypeptides having physiological activity such as antibodies can also be exemplified. Nutritional substances such as vitamins can also be exemplified. Examples also include pharmacological agents such as antibiotics, hormones, and interferons. Other examples include natural pigments for imparting a certain color to the target animal. Examples thereof include minerals such as organic or inorganic calcium, iron, and magnesium.

〔対象動物〕
本発明に係る生理活性物質の投与方法の対象となる動物は、消化管における油球吸収作用が認められている非ヒト動物である限りにおいて限定されない。特に好ましくは、水棲動物又は非ヒト陸棲哺乳動物を例示することができる。
[Target animal]
The animal that is the target of the method for administering a physiologically active substance according to the present invention is not limited as long as it is a non-human animal that is recognized to absorb oil balls in the digestive tract. Particularly preferably, aquatic animals or non-human terrestrial mammals can be exemplified.

より具体的には、消化管における油球吸収作用が認められる限りにおいて、例えばウサギ、マウス、イヌ、ネコ、ウシ、ブタ等の非ヒト哺乳動物、鳥類、両生類、爬虫類、養殖魚や鑑賞魚を含む各種の魚類、貝類、水棲甲殻類(エビ、カニ等)等を好ましく例示することができる。   More specifically, as long as an oil-ball absorbing action in the gastrointestinal tract is recognized, for example, non-human mammals such as rabbits, mice, dogs, cats, cows and pigs, birds, amphibians, reptiles, cultured fish and appreciation fish are included. Various kinds of fish, shellfish, oyster shellfish (shrimp, crab, etc.) can be preferably exemplified.

〔第1発明〜第6発明に係る乳化液〕
第1発明〜第6発明に係る乳化液とは、「水中油滴型の乳化液であって、その油滴が生理活性物質を含むと共に動物の消化管における油球吸収作用の対象となる平均粒径を有する微小油球であるもの」を言う。
[Emulsions according to the first to sixth inventions]
The emulsions according to the first to sixth inventions are “an oil-in-water type emulsion, wherein the oil droplets contain a physiologically active substance and are targeted for oil ball absorption in the digestive tract of animals. “It is a fine oil sphere having a particle size”.

ここに、微小油球における「油球吸収作用の対象となる平均粒径」は、対象動物の種類に対応して、一律には限定できないが、例えば1〜60μmの範囲内の平均粒径が好ましく、1〜20μmの範囲内の平均粒径が更に好ましい。微小油球の粒度分布(各微小油球の粒揃いの程度)も限定されないが、上記の平均粒径の範囲内において、なるべく粒度分布の狭いものが好ましい。   Here, the “average particle size that is the target of the oil ball absorption action” in the micro oil spheres is not limited to a uniform size according to the type of the target animal, but for example, the average particle size in the range of 1 to 60 μm An average particle diameter within the range of 1 to 20 μm is more preferable. The particle size distribution of the fine oil spheres (the degree of particle alignment of each fine oil sphere) is not limited, but those having as narrow a particle size distribution as possible within the above average particle size range are preferable.

微小油球の具体的な粒子構造は、油球吸収作用の対象となる限りにおいて限定されないが、以下に述べる(1)〜(3)の3種類の粒子構造を好ましく例示することができる。
(1)微小油球中において、油溶性乳化剤の作用により、生理活性物質がそのまま、あるいは少量の水分と共に、極めて微小な分散相を形成している。
(2)微小油球中において、親油性の生理活性物質が溶解又は懸濁している。
(3)微小油球が脂質二重層によって形成されたミセル構造を持ち、その内部に生理活性物質が封入されている。
The specific particle structure of the fine oil sphere is not limited as long as it is an object of oil ball absorption, but the following three types of particle structures (1) to (3) can be preferably exemplified.
(1) In the fine oil sphere, the bioactive substance forms an extremely fine dispersed phase as it is or with a small amount of water by the action of the oil-soluble emulsifier.
(2) A lipophilic physiologically active substance is dissolved or suspended in the fine oil sphere.
(3) A micro oil sphere has a micelle structure formed by a lipid bilayer, and a physiologically active substance is enclosed therein.

上記の内、(2)は、油液に親油性の生理活性物質を溶解又は微細に懸濁させ、この油液と水とを用いて(好ましくは更に乳化剤を用いて)水中油滴型の乳化液を構成するものである。又、(3)は、水中において、脂質二重層によって形成されたミセル構造(その内部に生理活性物質の水溶液等が封入されている)を構成するものである。これらの(2)と(3)との粒子構造の調製方法、あるいは、これらの粒子構造を持つ微小油球が分散した乳化液の調製方法は、公知又は周知であり、それらの公知又は周知の調製方法の内から適宜な方法を選択して、容易に実施することができる。(1)の粒子構造とその調製方法とは、以下の項において詳しく述べる。   Among the above, (2) is an oil-in-water droplet type wherein a lipophilic physiologically active substance is dissolved or finely suspended in an oil solution, and this oil solution and water are used (preferably further using an emulsifier). It constitutes an emulsion. Further, (3) constitutes a micelle structure (an aqueous solution of a physiologically active substance or the like is enclosed therein) formed by a lipid bilayer in water. The method for preparing the particle structure of (2) and (3) or the method for preparing the emulsion in which the fine oil spheres having these particle structures are dispersed is publicly known or well known. An appropriate method can be selected from the preparation methods and can be easily carried out. The particle structure of (1) and its preparation method will be described in detail in the following section.

〔生理活性物質含有乳化液〕
第7発明〜第9発明に係る生理活性物質含有乳化液においては、動物の消化管における油球吸収作用の対象となる平均粒径を有する微小油球が分散した水中油滴型の乳化液であって、該微小油球中においては、油溶性乳化剤の作用により、生理活性物質がそのまま、あるいは少量の水分と共に極めて微小な分散相を形成して油相中に包埋されている。
[Bioactive substance-containing emulsion]
The biologically active substance-containing emulsion according to the seventh to ninth inventions is an oil-in-water emulsion in which fine oil balls having an average particle size that are targets for oil ball absorption in the digestive tract of animals are dispersed. In the fine oil sphere, the bioactive substance is embedded in the oil phase as it is or with a small amount of water by forming an extremely fine dispersed phase by the action of the oil-soluble emulsifier.

ここにおいて、上記の微小油球中には、生理活性物質の(又は、生理活性物質と少量の水分との)極めて微小な分散相が、その個々の分散相における分散質との界面に油溶性乳化剤を保持した状態で、分散質たる微小油球の油相中に安定的に分散している。そして、このような微小油球が、その個々の微小油球における分散質(水)との界面に水溶性乳化剤を保持した状態で、分散質たる水中に安定的に分散している。   Here, in the micro oil spheres described above, an extremely minute dispersed phase of a physiologically active substance (or a physiologically active substance and a small amount of water) is oil-soluble at the interface with the dispersoid in each individual dispersed phase. It is stably dispersed in the oil phase of fine oil spheres as a dispersoid while retaining the emulsifier. Such fine oil spheres are stably dispersed in water as a dispersoid while holding a water-soluble emulsifier at the interface with the dispersoid (water) in each of the fine oil spheres.

生理活性物質は、その親和性が水溶性、親水性、油溶性、親油性のいずれであっても良い。微小油球の好ましい平均粒径や粒度分布については、上記した「第1発明〜第6発明に係る乳化液」の場合と同様である。   The affinity of the physiologically active substance may be any of water solubility, hydrophilicity, oil solubility, and lipophilicity. The preferable average particle size and particle size distribution of the fine oil spheres are the same as those of the above-mentioned “emulsions according to the first to sixth inventions”.

第7発明〜第9発明に係る生理活性物質含有乳化液に含まれる微小油球の特徴点は、油溶性乳化剤の作用により、微小油球中で生理活性物質が極めて微小な分散相を形成している(油相中に包埋されている)点にある。そして、生理活性物質はそのまま、あるいは少量の水分と共に微小分散しているため、微小な分散相における生理活性物質の濃度は、100%ないしは極めて高濃度であり、よってドラッグデリバリー効果が著しく高い。なお、生理活性物質のこのような高濃度かつ微小分散の状態は、次の項で述べる乳化液の製造方法によって実現可能となるものである。   The characteristic feature of the fine oil spheres contained in the bioactive substance-containing emulsion according to the seventh to ninth inventions is that the bioactive substance forms an extremely fine dispersed phase in the fine oil spheres by the action of the oil-soluble emulsifier. (Embedded in the oil phase). Since the physiologically active substance is finely dispersed as it is or with a small amount of water, the concentration of the physiologically active substance in the minute dispersed phase is 100% or extremely high, and thus the drug delivery effect is extremely high. Such a state of high concentration and fine dispersion of the physiologically active substance can be realized by the method for producing an emulsion described in the next section.

〔生理活性物質含有乳化液の製造方法〕
第10発明に係る生理活性物質含有乳化液の製造方法は、以下に順次述べるところの(A)第1乳化工程と、(B)第2乳化工程とを必須の工程として含む。但し、必要又は有益である限りにおいて、これらの工程に対して、任意の前処理工程、中間工程あるいは後処理工程を付加することもできる。
[Method of producing emulsion containing physiologically active substance]
The manufacturing method of the physiologically active substance-containing emulsion according to the tenth invention includes (A) a first emulsification step and (B) a second emulsification step as essential steps, which will be sequentially described below. However, as long as it is necessary or beneficial, an arbitrary pretreatment step, intermediate step or post-treatment step can be added to these steps.

(A)第1乳化工程:生理活性物質又はその水溶液もしくは水懸濁液と、油溶性乳化剤を添加した油液とを混合して乳化させることにより、生理活性物質がそのまま、あるいは少量の水分と共に極めて微小な分散相を形成して油相中に包埋されている油性乳化相を製造する。   (A) 1st emulsification process: By mixing and emulsifying the bioactive substance or its aqueous solution or water suspension and the oil liquid to which the oil-soluble emulsifier is added, the bioactive substance remains as it is or with a small amount of water. An oily emulsified phase is produced which forms a very fine dispersed phase and is embedded in the oil phase.

この第1乳化工程においては、生理活性物質をその水溶液もしくは水懸濁液として用いる場合には、水分をなるべく少なくすること、即ち、生理活性物質の高濃度水溶液あるいは高密度水懸濁液とすることが好ましい。   In this first emulsification step, when the physiologically active substance is used as an aqueous solution or aqueous suspension thereof, the water should be reduced as much as possible, that is, a highly concentrated aqueous solution or a high density aqueous suspension of the physiologically active substance. It is preferable.

第1乳化工程における重要な技術的ポイントは、生理活性物質やその水溶液等(a相)に対して油溶性乳化剤を添加した油液(b相)を混合して乳化させる点である。ここに、a相とb相の混合比率は一律には限定されないが、重量比で、例えばa相:b相=1:99〜99:1程度の範囲内で任意に選択される。   An important technical point in the first emulsification step is to mix and emulsify a bioactive substance or an aqueous solution thereof (phase a) with an oil solution (phase b) added with an oil-soluble emulsifier. Here, the mixing ratio of the a phase and the b phase is not uniformly limited, but is arbitrarily selected in a weight ratio, for example, within a range of about a phase: b phase = 1: 99 to 99: 1.

a相とb相の重量比が上記の範囲を逸脱すると、前記したような生理活性物質包埋型の微小油球の製造に失敗し易い。又、b相に油溶性乳化剤を添加しておかないと、前記したような生理活性物質包埋型の微小油球の製造に失敗し易い。b相に対する油溶性乳化剤の添加量は、1〜5重量%程度が適当である。   When the weight ratio of the a phase and the b phase deviates from the above range, the production of the bioactive substance-embedded micro oil sphere as described above tends to fail. If an oil-soluble emulsifier is not added to the b phase, the production of the bioactive substance-embedded micro oil spheres as described above tends to fail. The amount of the oil-soluble emulsifier added to the b phase is suitably about 1 to 5% by weight.

「油溶性乳化剤」とは、毒性や生理的安全性等に問題がない限りにおいて種類を限定されないが、そのHLB(親水性−親油性バランス)値が6以下のもの、特に2以下のものが好ましい。又、乳化剤のタイプとしては、非イオン性界面活性剤が好ましい。以上の条件に該当する好ましい油溶性乳化剤として、リノレイン酸ヘキサグリセリン、ポリオキシエチレンラウリルエーテル、レシチン、ポリオキシエチレンオレイルエーテル等を例示することができる。   “Oil-soluble emulsifier” is not limited in kind as long as there is no problem in toxicity, physiological safety, etc., but its HLB (hydrophilic-lipophilic balance) value is 6 or less, particularly 2 or less. preferable. As the type of emulsifier, a nonionic surfactant is preferable. Examples of preferable oil-soluble emulsifiers that meet the above conditions include linolenic acid hexaglycerin, polyoxyethylene lauryl ether, lecithin, polyoxyethylene oleyl ether, and the like.

(B)第2乳化工程:第1乳化工程で製造された油性乳化相と、水溶性乳化剤を添加した水とを混合して乳化させることにより、前記油性乳化相が動物の消化管における油球吸収作用の対象となる平均粒径を持つ微小油球として水中に分散した水中油滴型の乳化液を製造する。   (B) Second emulsification step: The oily emulsified phase produced in the first emulsification step and water added with a water-soluble emulsifier are mixed and emulsified, whereby the oily emulsified phase becomes an oil ball in the digestive tract of animals. An oil-in-water emulsion that is dispersed in water as fine oil spheres having an average particle size to be absorbed is produced.

第2乳化工程における技術的ポイントは、第1乳化工程で製造された油性乳化相(c相)に対して水溶性乳化剤を添加した水(d相)を混合して乳化させる点である。ここにc相とd相との混合比率は一律には限定されないが、重量比で、例えばc相:d相=2:1〜1:3程度のの範囲内で任意に選択される。   The technical point in a 2nd emulsification process is a point which mixes and emulsifies the water (d phase) which added the water-soluble emulsifier with the oil-based emulsification phase (c phase) manufactured at the 1st emulsification process. Here, the mixing ratio of the c phase and the d phase is not uniformly limited, but is arbitrarily selected in a weight ratio, for example, within a range of about c phase: d phase = 2: 1 to 1: 3.

c相とd相の重量比が上記の範囲を逸脱すると、本発明に係る生理活性物質含有乳化液の製造に失敗し易い。又、d相に水溶性乳化剤を添加しておかないと、同様に、本発明に係る生理活性物質含有乳化液の製造に失敗し易い。d相に対する水溶性乳化剤の添加量は、0.5〜1.3重量%程度が適当である。   If the weight ratio of the c phase and the d phase deviates from the above range, the production of the physiologically active substance-containing emulsion according to the present invention tends to fail. Similarly, unless a water-soluble emulsifier is added to the d phase, the production of the bioactive substance-containing emulsion according to the present invention is likely to fail. The amount of the water-soluble emulsifier added to the d phase is suitably about 0.5 to 1.3% by weight.

「水溶性乳化剤」とは、毒性や生理的安全性等に問題がない限りにおいて種類を限定されないが、そのHLB(親水性−親油性バランス)値が10以上のもの、特に12以上のものが好ましい。又、乳化剤のタイプとしては、非イオン性界面活性剤が好ましい。以上の条件に該当する好ましい水溶性乳化剤として、ジステアリン酸デカグリセリン、イソステアリン酸グリセリル、モノイソステアリン酸エチレングリコール、モノオレイン酸エチレングリコール等を例示することができる。   The “water-soluble emulsifier” is not limited as long as there is no problem in toxicity, physiological safety, etc., but its HLB (hydrophilic-lipophilic balance) value is 10 or more, particularly 12 or more. preferable. As the type of emulsifier, a nonionic surfactant is preferable. Examples of preferable water-soluble emulsifiers that meet the above conditions include decaglyceryl distearate, glyceryl isostearate, ethylene glycol monoisostearate, ethylene glycol monooleate, and the like.

以上の(A)第1乳化工程及び(B)第2乳化工程における乳化操作そのものは、公知の超音波を利用する方法や、ホモジナイザーないしはミキサー等を利用して適宜に行うことができる。それらの乳化操作の際、乳化手段の運転条件を適宜に選択することにより、生理活性物質含有乳化液における微小油球の粒径や、この微小油球中における生理活性物質の微細な分散の度合いを調整することができる。   The emulsification operation itself in the above (A) first emulsification step and (B) second emulsification step can be appropriately performed using a method using a known ultrasonic wave, a homogenizer, a mixer, or the like. During the emulsification operation, by appropriately selecting the operating conditions of the emulsification means, the particle size of the fine oil spheres in the emulsion containing the bioactive substance and the degree of fine dispersion of the bioactive substance in the fine oil spheres Can be adjusted.

(実施例1)
生理活性物質として、大腸菌のホルマリン死菌を用い、この死菌を高密度に含む水懸濁液(第1液)を調製した。一方、イカ肝油に対して油溶性乳化剤(リノレイン酸ヘキサグリセリン:商品名「サンソフト#818R」、太陽化学社製)を5重量%添加した油液(第2液)を調製した。
(Example 1)
Using a formalin killed Escherichia coli as a physiologically active substance, an aqueous suspension (first solution) containing the killed bacteria at a high density was prepared. On the other hand, an oil liquid (second liquid) was prepared by adding 5% by weight of an oil-soluble emulsifier (hexaglycerin linolenate: trade name “Sunsoft # 818R”, Taiyo Kagaku Co., Ltd.) to squid liver oil.

次に、前記第10発明に係る第1乳化処理として、第1液と第2液とを90:10の重量比で混合し、超音波発生器により十分な乳化処理を行って、大腸菌のホルマリン死菌の高密度懸濁液が極めて微小な分散相を形成して油相中に包埋されている油性乳化相(第3液)を形成した。   Next, as the first emulsification treatment according to the tenth aspect of the present invention, the first liquid and the second liquid are mixed at a weight ratio of 90:10, and sufficient emulsification treatment is performed with an ultrasonic generator to obtain E. coli formalin. The dense suspension of dead bacteria formed an extremely fine dispersed phase to form an oily emulsified phase (third liquid) embedded in the oil phase.

次に、水溶性乳化剤(ジステアリン酸デカグリセリン:商品名「サンソフト
Q−182S」、太陽化学社製)を1重量%添加した水(第4液)を調製した。そして、前記第10発明に係る第2乳化処理として、第3液と第4液とを2:1の重量比で混合し、ミキサーで十分に攪拌して、上記の油性乳化相が微小油球として水中に分散した生理活性物質含有乳化液を調製した。
Next, water (fourth liquid) to which 1% by weight of a water-soluble emulsifier (decaglyceryl distearate: trade name “Sunsoft Q-182S”, manufactured by Taiyo Chemical Co., Ltd.) was added was prepared. Then, as the second emulsification treatment according to the tenth aspect of the invention, the third liquid and the fourth liquid are mixed at a weight ratio of 2: 1 and sufficiently stirred with a mixer, so that the oily emulsified phase is a fine oil sphere. A bioactive substance-containing emulsion dispersed in water was prepared.

この生理活性物質含有乳化液に含まれている微小油球の粒径を観察したところ、概ね5〜10μm程度であった。調製した生理活性物質含有乳化液を冷蔵庫中に約1週間保存し、その後再度、微小油球の粒径を観察したところ、やはり概ね5〜10μm程度であって、微小油球の分散状態の安定性を確認した。   When the particle size of the fine oil spheres contained in this physiologically active substance-containing emulsion was observed, it was about 5 to 10 μm. The prepared emulsion containing the physiologically active substance is stored in the refrigerator for about one week, and then the particle size of the micro oil spheres is observed again. It is about 5 to 10 μm, and the dispersion state of the micro oil spheres is stable. The sex was confirmed.

(実施例2)
本実施例ではヒラメの病原細菌である Edwardsiella tarda のホルマリン死菌を用いた。そして、この死菌を10個体/ml含む水懸濁液(第1液)を調製し、それ以外の点は実施例1と全く同様にして、生理活性物質含有乳化液を調製した。
(Example 2)
In this example, killed formalin of Edwardsiella tarda, a pathogenic bacterium of flounder, was used. Then, an aqueous suspension (first liquid) containing 10 7 individuals / ml of the dead bacteria was prepared, and a physiologically active substance-containing emulsion was prepared in exactly the same manner as in Example 1 except for that point.

次に、この生理活性物質含有乳化液を、市販の魚類飼料用の多孔質ペレットに適量含浸させた。そして魚体重100g当たりの1日の乳化液摂取量が10μlとなるように、15尾のヒラメに対して上記した多孔質ペレットを3日間連続摂餌させた。   Next, an appropriate amount of this physiologically active substance-containing emulsion was impregnated into a commercially available porous pellet for fish feed. Then, the above-described porous pellets were fed continuously for 15 days to 15 flounder so that the daily emulsion intake per 100 g of fish weight was 10 μl.

上記の摂餌期間の終了後23日目に、上記のヒラメのうち5尾の尾動脈から採血し、血清中の抗 Edwardsiella tarda 抗体の抗体価を凝集価として測定した。一方、比較例として、生理活性物質含有乳化液を摂餌させていないヒラメ5尾の尾動脈から採血し、同様に血清中の抗 Edwardsiella tarda 抗体の抗体価を凝集価として測定した。   On the 23rd day after the end of the feeding period, blood was collected from 5 tail arteries of the flounder, and the antibody titer of anti-Edwardsiella tarda antibody in the serum was measured as the aggregation titer. On the other hand, as a comparative example, blood was collected from five flounder tail arteries not fed with a physiologically active substance-containing emulsion, and the antibody titer of anti-Edwardsiella tarda antibody in the serum was similarly measured as an agglutination titer.

それらの測定結果を以下に示す。以下において、「投与区」が実施例に係る5尾を、「対照区」が比較例に係る5尾を示す。又、「魚」の欄の数字は、投与区及び対照区の5尾のコイの検体番号である。   The measurement results are shown below. In the following, “administration group” shows 5 fishes according to the example, and “control group” shows 5 fishes according to the comparative example. The numbers in the “Fish” column are specimen numbers of five carp in the administration group and the control group.

魚 投与区凝集価 対照区凝集価
1 1024 2
2 1024 4
3 256 2
4 32 16
5 64 16
上記の凝集価測定とは別に、本実施例に係る摂餌期間の終了後23日目の前記15尾のヒラメのうち、残りの10尾に対しては、 Edwardsiella tarda 生菌を筋肉内に注射(攻撃菌量としては10個体/mlの菌液を0.3ml注射)して14日間飼育し、生存率を評価した。一方、比較例として、生理活性物質含有乳化液を摂餌させていないヒラメ10尾に対して同様に Edwardsiella tarda 生菌を筋肉内に注射して14日間飼育し、生存率を評価した。
Fish Administration group agglutination value Control group agglutination value 1 1024 2
2 1024 4
3 256 2
4 32 16
5 64 16
Apart from the agglutination titer measurement described above, the remaining 10 flounder 23 days after the end of the feeding period according to this example were injected intramuscularly with viable Edwardsiella tarda. (As the amount of attacking bacteria, 0.3 ml of 10 5 individuals / ml of the bacterial solution was injected) and reared for 14 days, and the survival rate was evaluated. On the other hand, as a comparative example, live Edwardsiella tarda was similarly injected into 10 muscles of flounder that were not fed with a biologically active substance-containing emulsion, and were reared for 14 days to evaluate the survival rate.

それらの評価結果を以下に示す。以下において「経過日数」の欄の数字は生菌注射後の経過日数を示す。又、「投与区」が実施例に係るヒラメを、「対照区」が比較例に係るヒラメを示し、各区の数字はその日に死亡した個体数を示す。投与区、対照区ともに、経過日数6日目までの死亡個体はなかった。対照区は14日経過時点での死亡個体数累計が10(生存率0%)であったのに対し、投与区は14日経過時点で死亡個体数累計が5(生存率50%)であった。   The evaluation results are shown below. In the following, the number in the “Elapsed days” column indicates the number of days that have elapsed since the injection of viable bacteria. In addition, “administration group” indicates the flounder according to the example, “control group” indicates the flounder according to the comparative example, and the numbers in each group indicate the number of individuals who died on that day. There were no dead animals until the 6th day in the treatment group and the control group. In the control plot, the cumulative number of dead individuals was 10 (survival rate 0%) at 14 days, whereas in the treated plot, the cumulative dead population was 5 (survival rate 50%) after 14 days. It was.

経過日数 投与区 対照区
7 0 0
8 0 0
9 0 1
10 0 4
11 1 1
12 1 0
13 3 1
14 0 3
以上の結果から分かるように、抗体価(凝集価)の測定においても、生菌注射後の生存率においても、投与区は対照区に比べて顕著に優れた結果を示した。
Elapsed days Administration group Control group 7 0 0
8 0 0
9 0 1
10 0 4
11 1 1
12 1 0
13 3 1
14 0 3
As can be seen from the above results, in the measurement of the antibody titer (aggregation titer) and the survival rate after injection of viable bacteria, the administration group showed significantly superior results as compared with the control group.

(実施例3)
本実施例でも、上記 Edwardsiella tarda のホルマリン死菌を用いた。そして、この死菌を10個体/ml含む水懸濁液(第1液)を調製する一方、実施例1の場合と同じ第2液を調製した。
(Example 3)
In the present example, the formalin killed by Edwardsiella tarda was also used. Then, while preparing an aqueous suspension (first liquid) containing 10 9 individuals / ml of the dead bacteria, the same second liquid as in Example 1 was prepared.

次に、前記第10発明に係る第1乳化処理として、第1液と第2液とを99:1の重量比で混合し、超音波発生器により十分な乳化処理を行って、大腸菌のホルマリン死菌の高密度懸濁液が極めて微小な分散相を形成して油相中に包埋されている油性乳化相(グループ1の第3液)を形成した。又、上記の第1液と第2液とを90:10の重量比で混合し、同上の乳化処理を行って油性乳化相(グループ2の第3液)を形成した。   Next, as the first emulsification treatment according to the tenth aspect of the present invention, the first liquid and the second liquid are mixed at a weight ratio of 99: 1, and sufficient emulsification treatment is performed with an ultrasonic generator to obtain E. coli formalin. The dense suspension of dead bacteria formed a very fine dispersed phase to form an oily emulsified phase (Group 1 third liquid) embedded in the oil phase. Moreover, said 1st liquid and 2nd liquid were mixed by the weight ratio of 90:10, and the same emulsification process was performed, and the oil-based emulsification phase (3rd liquid of group 2) was formed.

次に、実施例1と同じ内容の第4液を調製し、前記第10発明に係る第2乳化処理として、グループ1の第3液及びグループ2の第3液についてそれぞれ、第3液と第4液とを2:1の重量比で混合し、ミキサーで十分に攪拌して、上記の油性乳化相が微小油球として水中に分散した生理活性物質含有乳化液(「グループ1の生理活性物質含有乳化液」及び「グループ2の生理活性物質含有乳化液」)を調製した。   Next, the 4th liquid of the same content as Example 1 is prepared, and as the 2nd emulsification processing concerning the 10th invention, about the 3rd liquid of group 1 and the 3rd liquid of group 2, respectively, 4 liquids were mixed at a weight ratio of 2: 1, sufficiently stirred with a mixer, and the above-mentioned oily emulsified phase was dispersed in water as fine oil balls ("Group 1 physiologically active substances" Containing emulsions "and" group 2 physiologically active substance-containing emulsions ").

次に、グループ1の生理活性物質含有乳化液とグループ2の生理活性物質含有乳化液とを、それぞれ、市販の魚類飼料用の多孔質ペレットに適量含浸させた。そしてこれらの多孔質ペレットを、魚体重100g当たりの1日の乳化液摂取量が0.05mlとなるように、それぞれ10尾のヒラメ(平均体重は約100g)に対して3日間自由摂餌させた。グループ1の生理活性物質含有乳化液を摂餌したヒラメを「グループ1」、グループ2の生理活性物質含有乳化液を摂餌したヒラメを「グループ2」と呼ぶ。   Next, a commercially available porous pellet for fish feed was impregnated in an appropriate amount with the group 1 physiologically active substance-containing emulsion and the group 2 physiologically active substance-containing emulsion. These porous pellets were allowed to feed freely for 10 days for each 10 flounder (average body weight is about 100 g) so that the daily emulsion intake per 100 g of fish weight was 0.05 ml. It was. The flounder that feeds the group 1 bioactive substance-containing emulsion is called “Group 1”, and the flounder that feeds the group 2 bioactive substance-containing emulsion is called “Group 2”.

次に「グループ1」及び「グループ2」に係るヒラメについて、摂餌期間の終了後23日目にヒラメの血清中の抗 Edwardsiella tarda 抗体の抗体価を凝集価として測定した。その結果を表1の「菌体包埋油球投与区」の欄に示す。表1の「魚」欄の数字は「グループ1( Group-1)」及び「グループ2( Group-2)」に係る10尾のヒラメの検体番号である。   Next, for the flounder related to “Group 1” and “Group 2”, the antibody titer of the anti-Edwardsiella tarda antibody in the serum of the flounder was measured as the aggregation titer on the 23rd day after the end of the feeding period. The results are shown in the column of “Bacteria embedded in oil spheres” in Table 1. The numbers in the “Fish” column of Table 1 are the specimen numbers of 10 flounder samples related to “Group 1” and “Group 2”.

表1から分かるように、「グループ1」及び「グループ2」のヒラメにおいては、全体的に高濃度の抗 Edwardsiella tarda 抗体が産生されていた。   As can be seen from Table 1, in the flounder of “Group 1” and “Group 2”, a high concentration of anti-Edwardsiella tarda antibody was produced overall.

(実施例3に対する比較例)
実施例3と同じ Edwardsiella tarda のホルマリン死菌と、イカ肝油と、油溶性乳化剤とを、実施例3の「グループ1」の場合と同量に用いて単に混和しただけの油性液を調製した。
(Comparative example with respect to Example 3)
The same Edwardsiella tarda formalin killing bacteria as in Example 3, squid liver oil, and an oil-soluble emulsifier were used in the same amount as in “Group 1” in Example 3 to prepare an oily solution.

この油性液は、第10発明に係る第1乳化処理の結果物たる油性乳化相となっている可能性はあるが、第2乳化処理に相当するプロセスを経ていないため、本発明に係る生理活性物質含有乳化液にはなっていない。   Although this oily liquid may be an oily emulsified phase as a result of the first emulsification treatment according to the tenth invention, it has not undergone a process corresponding to the second emulsification treatment, so the physiological activity according to the present invention It is not a substance-containing emulsion.

この油性液を実施例3と同様に多孔質ペレットに含浸させ、実施例3と同様に10尾のヒラメに投与して抗体産生の評価を行った。この油性液を摂餌したヒラメを「グループ4( Group-4)」のヒラメ群とし、その評価結果を表1の「対象区」中の「 Group-4」の欄に示す。更に、 Edwardsiella tarda のホルマリン死菌や乳化液等を如何なる形態でも投与せず、全く通常に飼養している10尾のヒラメを「グループ3( Group-3)」のヒラメ群とし、これらのヒラメについても、同様の方法で抗体産生の評価を行った。その評価結果を表1の「対象区」中の「 Group-3」の欄に示す。   The oily liquid was impregnated into porous pellets in the same manner as in Example 3 and administered to 10 flounder in the same manner as in Example 3 to evaluate antibody production. The flounder fed with this oily liquid is defined as the “Group 4” flounder group, and the evaluation results are shown in the “Group-4” column of “Target Area” in Table 1. In addition, 10 groups of flounder that are not normally administered with any formalin killed bacteria or emulsions of Edwardsiella tarda and are kept in a normal manner are designated as “Group 3” flounder groups. In the same manner, antibody production was evaluated in the same manner. The evaluation results are shown in the “Group-3” column of “Target Area” in Table 1.

表1から分かるように、「グループ3」と「グループ4」との評価結果の間には余り差異がなく、かつ、これらと「グループ1」及び「グループ2」の評価結果との顕著な差異が明瞭である。   As can be seen from Table 1, there is not much difference between the evaluation results of “Group 3” and “Group 4”, and a significant difference between these and the evaluation results of “Group 1” and “Group 2” Is clear.

Figure 2005119986
Figure 2005119986

本願発明により、例えば養殖用又は鑑賞用の各種の魚類や水棲甲殻類等に対して、あるいは油球吸収作用が確認されている哺乳動物等に対して、免疫賦活物質や薬理活性物質その他の各種の生理活性物質を、簡易な手段によって非常に高い効率で摂取させる経口的ドラッグデリバリー手段として利用できる。
According to the invention of the present application, for example, various types of fish or aquatic shellfish for aquaculture or appreciation, or for mammals that have been confirmed to absorb oil balls, immunostimulatory substances, pharmacologically active substances, and other various types. The physiologically active substance can be used as an oral drug delivery means for taking in a very high efficiency by a simple means.

Claims (10)

水中油滴型の乳化液であって、その油滴が生理活性物質を含むと共に動物の消化管における油球吸収作用の対象となる平均粒径を有する微小油球であるものを調製し、
この乳化液を、消化管における油球吸収作用が認められている非ヒト動物に経口投与することを特徴とする生理活性物質の投与方法。
Prepare an oil-in-water emulsion, which is a micro oil sphere having an average particle size that contains a physiologically active substance and is subject to oil sphere absorption action in the digestive tract of animals,
A method for administering a physiologically active substance, wherein the emulsion is orally administered to a non-human animal that is recognized to absorb oil balls in the digestive tract.
前記微小油球が、1〜60μmの範囲内の平均粒径を有することを特徴とする請求項1に記載の生理活性物質の投与方法。 The method for administering a physiologically active substance according to claim 1, wherein the micro oil spheres have an average particle diameter in the range of 1 to 60 µm. 前記微小油球が以下(1)〜(3)のいずれかの粒子構造を持つものであることを特徴とする請求項1又は請求項2に記載の生理活性物質の投与方法。
(1)微小油球中において、油溶性乳化剤の作用により、生理活性物質がそのまま、あるいは少量の水分と共に、極めて微小な分散相を形成している。
(2)微小油球中において、親油性の生理活性物質が溶解又は懸濁している。
(3)微小油球が脂質二重層によって形成されたミセル構造を持ち、その内部に生理活性物質が封入されている。
The method for administering a physiologically active substance according to claim 1 or 2, wherein the micro oil sphere has a particle structure of any one of (1) to (3) below.
(1) In the fine oil sphere, the bioactive substance forms an extremely fine dispersed phase as it is or with a small amount of water by the action of the oil-soluble emulsifier.
(2) A lipophilic physiologically active substance is dissolved or suspended in the fine oil sphere.
(3) A micro oil sphere has a micelle structure formed by a lipid bilayer, and a physiologically active substance is enclosed therein.
前記乳化液を、飼料又は飼料用ペレットに含浸させたもとで動物に摂食させることを特徴とする請求項1〜請求項3のいずれかに記載の生理活性物質の投与方法。 The method for administering a physiologically active substance according to any one of claims 1 to 3, wherein the animal is fed while the emulsion is impregnated with feed or feed pellets. 前記動物が、水棲動物又は非ヒト陸棲哺乳動物であることを特徴とする請求項1〜請求項4のいずれかに記載の生理活性物質の投与方法。 The method for administering a physiologically active substance according to any one of claims 1 to 4, wherein the animal is an aquatic animal or a non-human terrestrial mammal. 前記生理活性物質が、ワクチン、免疫賦活物質、栄養物質、薬理作用物質、天然色素又はミネラルであることを特徴とする請求項1〜請求項5のいずれかに記載の生理活性物質の投与方法。 The method for administering a physiologically active substance according to any one of claims 1 to 5, wherein the physiologically active substance is a vaccine, an immunostimulatory substance, a nutrient substance, a pharmacologically active substance, a natural pigment or a mineral. 動物の消化管における油球吸収作用の対象となる平均粒径を有する微小油球が分散した水中油滴型の乳化液であって、該微小油球中においては、油溶性乳化剤の作用により、生理活性物質がそのまま、あるいは少量の水分と共に極めて微小な分散相を形成して油相中に包埋されていることを特徴とする生理活性物質含有乳化液。 An oil-in-water emulsion in which fine oil spheres having an average particle size to be absorbed in the animal's gastrointestinal tract are dispersed. In the fine oil spheres, due to the action of an oil-soluble emulsifier, A bioactive substance-containing emulsion characterized in that a bioactive substance is embedded in an oil phase as it is or with a small amount of water to form a very fine dispersed phase. 前記微小油球が、1〜60μmの範囲内の平均粒径を有することを特徴とする請求項7に記載の生理活性物質含有乳化液。 8. The bioactive substance-containing emulsion according to claim 7, wherein the fine oil spheres have an average particle diameter in the range of 1 to 60 [mu] m. 前記生理活性物質が、ワクチン、免疫賦活物質、栄養物質、薬理作用物質、天然色素又はミネラルであることを特徴とする請求項7又は請求項8に記載の生理活性物質含有乳化液。 9. The physiologically active substance-containing emulsion according to claim 7 or 8, wherein the physiologically active substance is a vaccine, an immunostimulatory substance, a nutrient substance, a pharmacologically active substance, a natural pigment or a mineral. 以下の(A)第1乳化工程と(B)第2乳化工程とを含むことを特徴とする生理活性物質含有乳化液の製造方法。
(A)第1乳化工程:生理活性物質又はその水溶液もしくは水懸濁液と、油溶性乳化剤を添加した油液とを混合して乳化させることにより、生理活性物質がそのまま、あるいは少量の水分と共に極めて微小な分散相を形成して油相中に包埋されている油性乳化相を製造する。
(B)第2乳化工程:第1乳化工程で製造された油性乳化相と、水溶性乳化剤を添加した水とを混合して乳化させることにより、前記油性乳化相が動物の消化管における油球吸収作用の対象となる平均粒径を持つ微小油球として水中に分散した水中油滴型の乳化液を製造する。
The manufacturing method of the bioactive substance containing emulsion characterized by including the following (A) 1st emulsification processes and (B) 2nd emulsification processes.
(A) 1st emulsification process: By mixing and emulsifying the bioactive substance or its aqueous solution or water suspension and the oil liquid to which the oil-soluble emulsifier is added, the bioactive substance remains as it is or with a small amount of water. An oily emulsified phase is produced which forms a very fine dispersed phase and is embedded in the oil phase.
(B) Second emulsification step: The oily emulsified phase produced in the first emulsification step and water added with a water-soluble emulsifier are mixed and emulsified, whereby the oily emulsified phase becomes an oil ball in the digestive tract of animals. An oil-in-water emulsion that is dispersed in water as fine oil spheres having an average particle size to be absorbed is produced.
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