JPH06166635A - Immunoadjuvant - Google Patents
ImmunoadjuvantInfo
- Publication number
- JPH06166635A JPH06166635A JP18596892A JP18596892A JPH06166635A JP H06166635 A JPH06166635 A JP H06166635A JP 18596892 A JP18596892 A JP 18596892A JP 18596892 A JP18596892 A JP 18596892A JP H06166635 A JPH06166635 A JP H06166635A
- Authority
- JP
- Japan
- Prior art keywords
- chitin
- water
- immunoadjuvant
- soluble
- chitosan
- 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.)
- Pending
Links
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明は、免疫アジュバントに関
するものである。詳しくは、本発明は、生体内に注入し
た抗原の、免疫効果を助成する免疫アジュバントに関す
るものである。FIELD OF THE INVENTION The present invention relates to an immunoadjuvant. More specifically, the present invention relates to an immunoadjuvant that promotes an immune effect of an antigen injected into a living body.
【０００２】免疫とは、生体が自己又は非自己（抗原）
を認識し、抗体を産生して非自己を処理排除するという
生体に備わった機能である。ある病原微生物に対する免
疫を付与する目的で病原性の低下した微生物、あるいは
不活化した病原微生物成分（ワクチン）の接種が実施さ
れる。この様な場合、有効な免疫応答を、目的とする局
所に惹起するために各種アジュバントが使用されてい
る。Immunity means that the living body is self or non-self (antigen)
Is a function of the body that recognizes and produces antibodies to process and eliminate non-self. For the purpose of immunizing a certain pathogenic microorganism, inoculation of a pathogenic microorganism or an inactivated pathogenic microorganism component (vaccine) is carried out. In such cases, various adjuvants have been used to elicit an effective immune response locally at the target.
【０００３】[0003]
【従来の技術】従来からキチンオリゴマー及びキトサン
オリゴマーにアジュバント活性があることが知られてい
る（特開昭62-61927号公報）。2. Description of the Related Art It has been conventionally known that chitin oligomers and chitosan oligomers have an adjuvant activity (JP-A-62-61927).
【０００４】[0004]
【発明が解決しようとする課題】しかしながら、キチン
オリゴマー、キトサンオリゴマーでは製造法に手間がか
かり、しかも効果が低い（有効投与量：50-150mg）とい
う欠点を有している。本発明は、キチン質にアジュバン
ト効果のあることを発見し、本発明を完成した。However, the chitin oligomer and chitosan oligomer have the drawbacks that the production method is troublesome and the effect is low (effective dose: 50-150 mg). The present invention discovered that chitin has an adjuvant effect and completed the present invention.
【０００５】本発明は、製造に手間がかからず、しかも
効果の高い、すなわち有効投与量の低いアジュバントの
提供を目的とするものである。It is an object of the present invention to provide an adjuvant which is easy to manufacture and has a high effect, that is, a low effective dose.
【０００６】[0006]
【課題を解決するための手段】本発明は、キチン質を有
効成分とする免疫アジュバントをその骨子とするもので
ある。The present invention has as its essence an immunoadjuvant containing chitin as an active ingredient.
【０００７】本発明に使用するキチン質としては、エ
ビ、カニ等の甲かく類、バッタ、カブトムシ等の昆虫
類、イカの甲等に含まれるキチン、キチンを20-80%脱ア
セチル化した脱アセチル化キチン、キトサン（特に脱ア
セチル化度80％以上のキチン）を挙げることができる。The chitin used in the present invention includes shrimp, crab and other shellfish, grasshopper, beetle and other insects, squid shell and other chitin, and deacetylated 20-80% of chitin. Examples include acetylated chitin and chitosan (particularly chitin having a deacetylation degree of 80% or more).
【０００８】本発明の免疫アジュバントに使用するキチ
ン質としては、水に不溶性、難溶性あるいは可溶性のキ
チン質を挙げることができる。水に不溶性及び難溶性の
キチン質としては、使用時、水に懸濁する程度に微粒子
化されたものが使用上推奨される。微粒子化の方法とし
ては、振動ボールミル、遊星ボールミル、遠心流動化ミ
ルなどのミルを用い、キチン質を乾式粉砕する方法等を
挙げることができる。さらに乾式粉砕方法に加えて、得
られた微粒子化キチン質を、水、生理食塩水等を用い、
湿式粉砕する方法を採用することも又使用上推奨され
る。本発明の水に不溶性あるいは難溶性のキチン質の粒
度としては、注射針をスムースに通過する程度の大きさ
を挙げることができる。水に可溶性のキチン質は、適当
な濃度に調整したものを使用すれば良い。Examples of chitins used in the immunoadjuvant of the present invention include water-insoluble, sparingly soluble or soluble chitins. As the chitin that is insoluble and hardly soluble in water, it is recommended to use fine particles that are finely dispersed to the extent that they are suspended in water during use. Examples of the method of forming fine particles include a method of dry-milling chitin using a mill such as a vibrating ball mill, a planetary ball mill, a centrifugal fluidizing mill. In addition to the dry pulverization method, the obtained finely divided chitin is used in water, physiological saline, etc.
It is also recommended to use the method of wet grinding. Examples of the particle size of the water-insoluble or sparingly soluble chitinous substance of the present invention include sizes that allow it to smoothly pass through an injection needle. The water-soluble chitin may be used after adjusting it to an appropriate concentration.
【０００９】本発明の免疫アジュバントの使用法は、ワ
クチンと混合した後注射する方法が挙げられる。また投
与量は10mg以下で十分効果を発揮する。Examples of the method of using the immunoadjuvant of the present invention include a method of injecting after mixing with a vaccine. A dose of 10 mg or less is sufficient to exert the effect.
【００１０】[0010]
【作用】本発明の免疫アジュバントがワクチン接種に効
果を発揮する理由は、今のところ明確には解明されてい
ないが、以下のごとく考えられる。生体内には、マクロ
ファージや多形核白血球に代表され、微生物などの異物
が生体内に進入したときにその異物を認識し、異物へ移
動し、異物を取り込み、そして酵素処理して排除すると
いう作用を持つ細胞、所謂貪食細胞が存在するが、キチ
ン質には、この貪食細胞を活性化させ、さらに活性化物
質に向かって濃度勾配に逆らって遊走する化学走化性を
も有している。キチン質をワクチンと共に注入すること
により注入部位に貪食細胞を遊走するとともに貪食作用
を活性化させ、抗原認識を助長することによって抗体産
生を活発化させるものと考えられる。またキトサンには
蛋白吸着能があることが知られており、ワクチン内の弱
毒化あるいは不活化ウイルスの吸着によって、より効率
よく抗原提示細胞（マクロファージ）に補食され易くな
るなどが総合してワクチン効果の加速化と抗体価の充分
な上昇が得られるものと考えられる。The reason why the immunoadjuvant of the present invention exerts an effect on vaccination has not yet been clarified yet, but it is considered as follows. Represented by macrophages and polymorphonuclear leukocytes in the body, when a foreign body such as a microorganism enters the body, it recognizes the foreign body, moves to the foreign body, takes in the foreign body, and removes it by enzymatic treatment. There are cells that act, so-called phagocytes, but the chitin also has chemotaxis that activates these phagocytes and migrates toward the activator against the concentration gradient. . It is considered that by injecting chitin together with a vaccine, phagocytic cells migrate to the injection site, activate phagocytosis, and promote antigen recognition, thereby activating antibody production. It is also known that chitosan has protein adsorption ability, and adsorption of attenuated or inactivated virus in the vaccine facilitates more efficient feeding by antigen-presenting cells (macrophages). It is considered that the effect is accelerated and the antibody titer is sufficiently increased.
【００１１】参考例１ カニのクチクラより製造したキトサン｛フローナック
Ｃ、脱アセチル化度83%（元素分析値より算出）、共和
テクノス製造｝を遠心流動化ミルを用い、乾式粉砕を行
った。粉砕条件はジルコニア製ボール（10mmφ、10kg使
用）を使用し、ミル回転数を420rpm、セパレーター回転
数を3500rpmで粉砕を行った。得られた粉末をSKレーサ゛ーPR
O-7000S（セイシン企業製造）によりエタノール中で測
定した結果、最大粒径12μm、平均粒径3.1μmであっ
た。Reference Example 1 Chitosan manufactured from crab cuticle {Flownac
C, deacetylation degree 83% (calculated from elemental analysis value, Kyowa Technos Production Co., Ltd.) was dry-ground using a centrifugal fluidizing mill. As the crushing conditions, balls made of zirconia (10 mmφ, 10 kg used) were used, and the crushing was performed at a mill rotation speed of 420 rpm and a separator rotation speed of 3500 rpm. The obtained powder is SK Laser PR
As a result of measurement in ethanol by O-7000S (manufactured by Seishin Enterprise Co., Ltd.), the maximum particle size was 12 μm and the average particle size was 3.1 μm.
【００１２】この粉末化キトサンを5%(w/w)の濃度で脱
イオン水に分散し5リットルの処理液を作成した。この
ときの粒径を粒度分析計（日機装株式会社製造、マイク
ロトラックSPA形）により測定した結果、温度11℃にお
いて平均粒径10.2μm、最大粒径42.2μmであった。This powdered chitosan was dispersed in deionized water at a concentration of 5% (w / w) to prepare a treatment liquid of 5 liters. The particle size at this time was measured by a particle size analyzer (manufactured by Nikkiso Co., Ltd., Microtrac SPA type). As a result, the average particle size was 10.2 μm and the maximum particle size was 42.2 μm at a temperature of 11 ° C.
【００１３】得られたキトサン分散液を攪拌下にサンド
グラインダーのベッセル内（容量 2リットル）に送り込
み、ジルコニア製ピン付羽根とジルコニア製ビーズ（0.
5mmφ、1.7リットル）と処理液とを強力に対流させるこ
とにより粉砕を行った。処理液はベッセル内を7回通過
させ、6ヶ月保存しても沈降することなく安定な分散性
を示し、針径0.4mm、針長20mmの皮下用注射針をスムー
ズに通過する微粒子状キトサン分散液を調製した。The resulting chitosan dispersion was fed into a vessel (capacity 2 liters) of a sand grinder with stirring, and a zirconia pin blade and zirconia beads (0.
(5 mmφ, 1.7 liter) and the treatment liquid were strongly convected to pulverize. The treatment solution passes through the vessel 7 times, shows stable dispersibility without sedimentation even after storage for 6 months, and fine particle chitosan dispersion that smoothly passes through a hypodermic injection needle with a needle diameter of 0.4 mm and a needle length of 20 mm. A liquid was prepared.
【００１４】[0014]
【実施例】以下実施例を挙げて更に詳しく説明するが、
本発明はかかる実施例に限定されるものではない。EXAMPLES The present invention will be described in more detail with reference to the following examples.
The present invention is not limited to such embodiments.
【００１５】実施例１ 参考例１で調整した微粒子状キトサン分散液をリンゲル
液で50μg/mlに希釈し、本発明の免疫アジュバントを製
造した。上記により製造した免疫アジュバント1mlとオ
ランダ、デュファー社製造ジステンパーワクチン1mlと
を約20分混和した後、2歳の雌雑種犬の皮下に投与し、
赤血球凝集反応法によって投与前と投与2週間後の抗体
価を測定した。その結果、接種前640であった抗体価が
接種2週間後5120以上に上昇した。Example 1 The fine particle chitosan dispersion prepared in Reference Example 1 was diluted to 50 μg / ml with Ringer's solution to prepare the immunoadjuvant of the present invention. The immunoadjuvant 1 ml produced by the above and the Netherlands, after mixing the Duffer Co. manufactured distemper vaccine 1 ml for about 20 minutes, subcutaneously administered to a 2-year-old female mongrel dog,
The antibody titer was measured before and 2 weeks after the administration by the hemagglutination method. As a result, the antibody titer, which was 640 before the inoculation, increased to 5120 or more 2 weeks after the inoculation.
【００１６】実施例２ カニのクチクラより製造したキトサンに代えて、イカ甲
より精製したキチンを使用した以外は参考例１と同様に
処理し、次いでリンゲル液でキチン粒子50μg／mlに希
釈して本発明の免疫アジュバントを製造した。このアジ
ュバント1mlとオランダ、デュファー社製造ヂステンパ
ーワクチン1mlとを約20分混和した後、3歳の雌ビーグル
犬の皮下に投与し、赤血球凝集反応法によって投与前と
投与2週間後の抗体価を測定した。その結果、接種前160
であった抗体価が接種2週間後5120以上に上昇した。Example 2 The same procedure as in Reference Example 1 was carried out except that chitin purified from squid shell was used instead of chitosan produced from cuticle of crab, and then diluted with Ringer's solution to 50 μg / ml of chitin particles. The immunoadjuvant of the invention was produced. 1 ml of this adjuvant and 1 ml of distemper vaccine manufactured by Duffer, Netherlands were mixed for about 20 minutes, and then subcutaneously administered to a 3-year-old female Beagle dog, and the antibody titer was measured before and 2 weeks after administration by the hemagglutination method. Was measured. As a result, before inoculation 160
The antibody titer, which was, increased to 5120 or more 2 weeks after the inoculation.
【００１７】比較例１ オランダ、デュファー社製造ヂステンパーワクチン1ml
を生後3ヶ月の雄ビーグル犬に接種し、赤血球凝集反応
法によって投与前と投与2週間後の抗体価を測定した。
その結果、接種前20であった抗体価が接種2週間後640で
あった。Comparative Example 1 1 ml of distemper vaccine manufactured by Duffer, Netherlands
Was inoculated into a 3-month-old male beagle dog, and the antibody titer was measured before and 2 weeks after the administration by hemagglutination method.
As a result, the antibody titer which was 20 before the inoculation was 640 2 weeks after the inoculation.
Claims (1)
ント。1. An immunoadjuvant containing chitin as an active ingredient.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP18596892A JPH06166635A (en) | 1992-06-18 | 1992-06-18 | Immunoadjuvant |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP18596892A JPH06166635A (en) | 1992-06-18 | 1992-06-18 | Immunoadjuvant |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH06166635A true JPH06166635A (en) | 1994-06-14 |
Family
ID=16180031
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP18596892A Pending JPH06166635A (en) | 1992-06-18 | 1992-06-18 | Immunoadjuvant |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH06166635A (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1996010421A1 (en) * | 1994-10-04 | 1996-04-11 | Medeva Holdings B.V. | Vaccine compositions |
| EP1027061A1 (en) * | 1997-10-03 | 2000-08-16 | Galenica Pharmaceuticals, Inc. | Imine-forming polysaccharides, preparation thereof and the use thereof as adjuvants and immunostimulants |
| EP1033995A1 (en) * | 1997-11-07 | 2000-09-13 | Protein Sciences Corporation | Insect cells or fractions as adjuvant for antigens |
| US6136606A (en) * | 1995-11-01 | 2000-10-24 | Medeva Holdings Bv | Influenza vaccine compositions |
| WO2000055345A3 (en) * | 1999-03-17 | 2001-05-31 | Univ Texas | Therapy of cancer by insect cells containing recombinant baculovirus encoding genes |
| US6316007B1 (en) | 1995-04-04 | 2001-11-13 | Wound Healing Of Oklahoma | Combined physical and immunotherapy for cancer |
| EP1574217A1 (en) * | 1997-10-03 | 2005-09-14 | Galenica Pharmaceuticals, Inc. | Imine-forming polysaccharides, preparation thereof and the use thereof as adjuvants and immunostimulants |
| US7074612B2 (en) * | 2002-03-06 | 2006-07-11 | National Institute Of Agrobiological Sciences | Insect cell primary culture medium, extracellular matrix, and process of preparing an insect culture cell line in a short period of time using the medium and matrix |
| JP2009541281A (en) * | 2006-06-20 | 2009-11-26 | シーエムピー セラピューティクス リミテッド | Compositions containing chitin microparticles and their medical use |
| JP2010511417A (en) | 2006-10-30 | 2010-04-15 | シーエムピー セラピューティクス リミテッド | Method for producing fine particles |
-
1992
- 1992-06-18 JP JP18596892A patent/JPH06166635A/en active Pending
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1996010421A1 (en) * | 1994-10-04 | 1996-04-11 | Medeva Holdings B.V. | Vaccine compositions |
| US6048536A (en) * | 1994-10-04 | 2000-04-11 | Medeva Holdings Bv | Vaccine compositions |
| US6316007B1 (en) | 1995-04-04 | 2001-11-13 | Wound Healing Of Oklahoma | Combined physical and immunotherapy for cancer |
| US6136606A (en) * | 1995-11-01 | 2000-10-24 | Medeva Holdings Bv | Influenza vaccine compositions |
| EP1027061A4 (en) * | 1997-10-03 | 2000-12-13 | Galenica Pharmaceuticals Inc | Imine-forming polysaccharides, preparation thereof and the use thereof as adjuvants and immunostimulants |
| EP1027061A1 (en) * | 1997-10-03 | 2000-08-16 | Galenica Pharmaceuticals, Inc. | Imine-forming polysaccharides, preparation thereof and the use thereof as adjuvants and immunostimulants |
| EP1574217A1 (en) * | 1997-10-03 | 2005-09-14 | Galenica Pharmaceuticals, Inc. | Imine-forming polysaccharides, preparation thereof and the use thereof as adjuvants and immunostimulants |
| US6960344B2 (en) | 1997-10-03 | 2005-11-01 | Galenica Pharmaceuticals, Inc. | Use of imine-forming polysaccharides as adjuvants and immunostimulants |
| US7196073B2 (en) | 1997-10-03 | 2007-03-27 | Adjuvantys, Inc. | Imine-forming polysaccharide adjuvants and immunostimulants |
| EP1033995A1 (en) * | 1997-11-07 | 2000-09-13 | Protein Sciences Corporation | Insect cells or fractions as adjuvant for antigens |
| EP1033995A4 (en) * | 1997-11-07 | 2001-01-10 | Protein Sciences Corp | Insect cells or fractions as adjuvant for antigens |
| EP1611897A3 (en) * | 1997-11-07 | 2007-08-15 | Protein Sciences Corporation | Whole or disrupted insect cells as adjuvant for antigens |
| WO2000055345A3 (en) * | 1999-03-17 | 2001-05-31 | Univ Texas | Therapy of cancer by insect cells containing recombinant baculovirus encoding genes |
| US7074612B2 (en) * | 2002-03-06 | 2006-07-11 | National Institute Of Agrobiological Sciences | Insect cell primary culture medium, extracellular matrix, and process of preparing an insect culture cell line in a short period of time using the medium and matrix |
| JP2009541281A (en) * | 2006-06-20 | 2009-11-26 | シーエムピー セラピューティクス リミテッド | Compositions containing chitin microparticles and their medical use |
| JP2010511417A (en) | 2006-10-30 | 2010-04-15 | シーエムピー セラピューティクス リミテッド | Method for producing fine particles |
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