JPS59210013A - Active substance-containing liposome - Google Patents
Active substance-containing liposomeInfo
- Publication number
- JPS59210013A JPS59210013A JP58078747A JP7874783A JPS59210013A JP S59210013 A JPS59210013 A JP S59210013A JP 58078747 A JP58078747 A JP 58078747A JP 7874783 A JP7874783 A JP 7874783A JP S59210013 A JPS59210013 A JP S59210013A
- Authority
- JP
- Japan
- Prior art keywords
- polysaccharide
- liposome
- suspension
- active substance
- glucosamine
- 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.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
- A61K9/1271—Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers
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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)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicinal Preparation (AREA)
Abstract
Description
【発明の詳細な説明】
本発明は、機械的強度が高く生体内での安全性の高い活
性物質含有新規リボン−１、に関する。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel ribbon-1 containing an active substance that has high mechanical strength and high safety in vivo.
リポソームはホスホリピドやコレステロール等のリン脂
質の２分子層よりなり、細胞膜と類似の組成をもつこと
から安全性の高い人工細胞を作るのに適しており、これ
を利用した人工血液のイ１ノ［究かなされている。Liposomes are composed of two molecular layers of phospholipids such as phospholipids and cholesterol, and have a similar composition to cell membranes, making them suitable for creating highly safe artificial cells. It has been investigated.
しかし、従来のリポソームは不安定で壊れ易く、例えば
ヘモグロビンのような蛋白質と結合させ、その中に封入
することか困難であった。また酸素運搬機能を有するヘ
モグロビンをリポソームに封入したとしてもその変形性
等の流動特性（レオロジー）についてはほとんど報告が
なされておらず、臨床的使用に耐えるか否か不明であっ
た。However, conventional liposomes are unstable and break easily, making it difficult to bind and encapsulate proteins such as hemoglobin. Furthermore, even if hemoglobin, which has an oxygen-carrying function, is encapsulated in liposomes, there have been few reports on its flow characteristics (rheology) such as deformability, and it was unclear whether the liposomes would be suitable for clinical use.
本発明者等は」−記問題に鑑みて鋭意イｉＪｆ究を重ね
た結果、リポソームの壁膜を形成するリン脂質の２分子
層にカルボキシメチルキチン、キトサン等のＤ−グルコ
サミンを主骨格成分とする多糖類を混合又は結合させる
ことにより機械的強度か高く、ヒツジヘモグロビンやヒ
トへモグロヒン等のは乳類動物の血液と同様の流動１、
コ性を示すリポソームが得られることを見い出した。The present inventors have conducted extensive research in view of the above problem, and have found that D-glucosamine, such as carboxymethyl chitin or chitosan, is added as a main skeleton component to the bimolecular layer of phospholipid that forms the wall of liposomes. Mechanical strength is increased by mixing or combining polysaccharides, and sheep hemoglobin and human hemoglobin have a similar flow rate as mammalian blood.
It has been found that liposomes exhibiting cohesive properties can be obtained.
即ち、本発明の特徴は、リン脂質の２分ｊ′層に機械的
強化剤としてＩ）−グルコサミンを主骨格成分とする多
糖類を混合又は結合させてなることを特徴とする活性物
質含有リポソームにある。That is, the present invention is characterized by an active substance-containing liposome comprising a bipartite layer of phospholipid mixed with or bonded with a polysaccharide whose main skeleton component is I)-glucosamine as a mechanical reinforcement. It is in.
本発明によれば、従来のリポソームに比べ機械的強度が
高いことか呟生体内外での安全性が高いのはもちろんカ
プセル化率か高く、更にカルボキシメチルで補強された
リポソームはそのカルボキシメチル基の存在により、化
学反応性を有し、細胞特異性をもたせるための化学修飾
をすることも可能である。According to the present invention, the mechanical strength of the liposomes is higher than that of conventional liposomes, and the encapsulation rate is high as well as the safety both inside and outside the body. Depending on their presence, they have chemical reactivity and can be chemically modified to provide cell specificity.
本発明の活性物質含有リポソームに使用するリン脂質と
しては、従来リポソームの壁膜形成に用いられている両
親媒性脂質であれば特に限定されず、例えばレシチン、
ケファリン、プラスマロゲン、α−グリセリルエーテル
、イ７シトールホスホリピッド、アミノアシルホスファ
チジルグリセロール、スフィンゴミリエン等か挙げられ
、これらを単独又は混合して、また必要に応してコレス
テロールやエルゴステロール等のステロール類ヲ添加す
るようにしてもよい。The phospholipids used in the active substance-containing liposomes of the present invention are not particularly limited as long as they are amphipathic lipids that have been conventionally used to form the wall of liposomes, such as lecithin,
Examples include cephalin, plasmalogen, α-glyceryl ether, isitol phospholipid, aminoacylphosphatidylglycerol, sphingomillien, etc., singly or in combination, and if necessary, sterols such as cholesterol and ergosterol. It may be added.
また、上記リン脂質に機械的強化剤として混合又は結合
さゼるＮ−アセチル−Ｄ−グルコサミンを主骨格成分と
する多糖類には、例えばキチンの誘導体であるカルボキ
シメチルキチン、カルホキジエチルキチン、それを加水
分解して得られるキトサン、カルボキシメチルキチン、
Ｎ−アセチル−Ｄ−グルコサミンとＤ−グルクロン酸と
か交互に結合したヒアルロン酸及びその誘導体等があり
、これらは単独或いは二種以」二を適宜組合せて用いる
ことができる。In addition, polysaccharides having N-acetyl-D-glucosamine as a main skeleton component that are mixed or bonded to the above-mentioned phospholipids as mechanical reinforcements include, for example, carboxymethyl chitin, carboxydiethyl chitin, which are derivatives of chitin, Chitosan, carboxymethyl chitin, obtained by hydrolyzing it,
There are hyaluronic acid and its derivatives in which N-acetyl-D-glucosamine and D-glucuronic acid are alternately bonded, and these can be used alone or in an appropriate combination of two or more.
」二記機械的強化剤としての多糖類を用いてリポソーム
を形成するに際しては、形成されるリポソームの壁膜中
に上記多糖類が０．０１乃至０．２重量％、好ましくは
０．０２乃至０．０５重量％存在するようにする。上記
壁膜中の多糖類の含量が０゜２重量％を越えると、合一
体となり、また、（）、０１重量％より低くなると機械
的強度の優れた安定なリポソームをＪＧる、二とができ
ない。2. When forming liposomes using polysaccharides as mechanical strengthening agents, the polysaccharides are present in the wall of the liposomes in an amount of 0.01 to 0.2% by weight, preferably 0.02 to 0.2% by weight. It should be present in an amount of 0.05% by weight. If the polysaccharide content in the wall membrane exceeds 0.2% by weight, it will coalesce, and if it is lower than 0.01% by weight, it will form a stable liposome with excellent mechanical strength. Can not.
一方、本発明のリポソームに１１人（包接）される活性
物質としては、人工血液の主体であるしツノヘモグロビ
ンやヒトヘモグロビン等のは乳類動物の赤血球、合成ヘ
モグロビン代替物をはじめとして、インシュリン、オキ
シトシン、バンプレシン、グルカゴン、フルチコトロピ
ン等のペプチドホルモン、黄体ホルモン、副腎ホルモン
等のステロイドホルモン、ペニシリン、ストレプトマイ
シン、クロラムフェニコール、バイオマイシン等の抗生
物質、ウレアーゼ、ペプシン、トリプシン、アルギナー
ゼ等の醇索、チアミン、ニコチン酸アミド、アスコルビ
ン酸等のビタミン類、その他５−ＦＵ（５−フルオロウ
ラシル）、マイトマイシン等の医薬か挙げられる。On the other hand, the active substances that are included in the liposomes of the present invention include mammalian red blood cells such as horn hemoglobin and human hemoglobin, which are the main components of artificial blood, synthetic hemoglobin substitutes, and insulin. , peptide hormones such as oxytocin, vamprecin, glucagon, and fluticotropin, steroid hormones such as progesterone and adrenal hormone, antibiotics such as penicillin, streptomycin, chloramphenicol, and biomycin, and urease, pepsin, trypsin, and arginase. Examples include vitamins such as thiamine, nicotinamide, and ascorbic acid, and pharmaceuticals such as 5-FU (5-fluorouracil) and mitomycin.
上記活性物質を包接した機械的強度の犬なる本発明のリ
ポソームを調製するには、レシチン等のリン脂質ヲジク
ロロメタン、クロロメタン、ベンゼン等の有機溶媒に溶
解しこれを上記ヒツジヘモグロビン等の活性物質の水溶
液と共に混合して激しく攪拌しＷ／○型のエマルジョン
とする。この場合、有機溶媒とリン脂質との使用割合は
、有機溶媒１００容量部に対してリン脂質１．０〜１０
．０重量部とするのが適当である。また、活性物質の水
溶液の濃度は５．０〜５０．０％（Ｗ／Ｖ）か゛適当で
ある。In order to prepare the mechanically strong liposome of the present invention containing the above-mentioned active substance, a phospholipid such as lecithin is dissolved in an organic solvent such as dichloromethane, chloromethane, benzene, etc. It is mixed with an aqueous solution of the substance and stirred vigorously to form a W/○ type emulsion. In this case, the ratio of organic solvent and phospholipid used is 1.0 to 10 parts by volume of phospholipid to 100 parts by volume of organic solvent.
．． It is appropriate to set the amount to 0 parts by weight. Further, the concentration of the aqueous solution of the active substance is suitably 5.0 to 50.0% (W/V).
このエマルジョンに上記機械的強化剤としての多糖類を
水性媒体に溶解又は分散させた液を加えて激しく攪拌し
Ｗ／○／Ｗ型のエマルジョンとした後、これを更に攪拌
を続けて上記有機）８媒を完全に蒸発させる。この多糖
類を溶解又は分散させる水性媒体としては、例えば、水
、生理食塩水、ｐＨ調整剤（水酸化ナトリウム、水酸化
カリウム）、緩衝剤（例えば、トリス−塩酸水溶液、リ
ン酸１ナトリウム、リン酸２ナトリウム水溶液）がある
。A solution prepared by dissolving or dispersing the above-mentioned polysaccharide as a mechanical strengthening agent in an aqueous medium is added to this emulsion and stirred vigorously to form a W/○/W type emulsion, which is then further stirred to form the above-mentioned organic) 8. Evaporate the medium completely. Examples of the aqueous medium in which this polysaccharide is dissolved or dispersed include water, physiological saline, pH adjusters (sodium hydroxide, potassium hydroxide), buffers (such as Tris-hydrochloric acid aqueous solution, monosodium phosphate, disodium acid aqueous solution).
この場合、多糖類の水性媒体中における濃度は、０．０
５〜０．５％（Ｗ／Ｖ）が適当であり、」上記Ｗ１０型
のエマルジョンへの添加に際しでは、全量を一度に添加
しても或いはこれを数回に分けて添加してもよい。なお
、これらの一連の操作は、室温で行い、特に活性物質か
ヒツジヘモグロビンやヒトヘモグロビンのように生体か
ら分ｇ＋ｒ、 Ｌ ？、＝ モのである場合には、窒素
等の不活性雰囲気で行うことが好ましい。In this case, the concentration of the polysaccharide in the aqueous medium is 0.0
5 to 0.5% (W/V) is appropriate, and when added to the above W10 type emulsion, the entire amount may be added at once or it may be added in several portions. Incidentally, these series of operations are performed at room temperature, and especially active substances such as sheep hemoglobin and human hemoglobin are separated from living organisms. , = Mo, it is preferable to carry out the reaction in an inert atmosphere such as nitrogen.
かくして、活性物質を包接しかつ多糖類によｉ）保護さ
れた安定なリポソームが分散したＷ／○／Ｗ型のエマル
ジョンか得られる。このリポソームは、粒子範囲が１４
０〜４２０ｎｍに集中し、平均経３１Ｏｒ＋ｍの球形で
あって毛細血管を自由に通過することが可能であり、２
０００〜１２０００ｇ、５〜３０分で遠心分離にかけ容
易に分離することかできる。従って、このエマルジョン
はこのまま使用することもできるが、上記遠心分離など
によりリポソームに封入されなかった活性物質を分離、
除去したのち、例えばヘモグロビン包接リポソームの場
合にはそのまま或いは要すれば他の栄養剤と共に輸血可
能な人工血液として、その他所望の注射液、経口剤、座
薬等の医薬や化粧剤として利用することができる。Thus, a W/O/W emulsion is obtained in which stable liposomes containing the active substance and protected i) by the polysaccharide are dispersed. This liposome has a particle range of 14
It is concentrated in the range of 0 to 420 nm, has a spherical shape with an average diameter of 31 Or+m, and can freely pass through capillaries.
It can be easily separated by centrifugation at 000 to 12,000 g for 5 to 30 minutes. Therefore, this emulsion can be used as is, but the active substance that was not encapsulated in the liposomes can be separated by the above-mentioned centrifugation, etc.
After removal, for example, in the case of a hemoglobin-included liposome, it can be used as an artificial blood that can be transfused as it is or with other nutritional supplements if necessary, or as other desired medicines or cosmetics such as injections, oral preparations, suppositories, etc. I can do it.
次に、本発明を実施例により具体的に説明する。Next, the present invention will be specifically explained using examples.
実施例１
カルボキシメチルキチン（ナンヨウ化成株製）を水に溶
解し０．２％（Ｗ／Ｖ）の溶液とした。デキス）・ラン
（Ｍｗ７３，３００ シグマ ケミカルカンパニー
セントルイス）をトリス−塩酸緩衝液（ｐＨ７，４）に
溶解し、６％（Ｗ／＼・“）の溶液とした。卵黄レシチ
ン（旭化成株製）を窒素気流中でン゛クロロメタンに溶
解し濃度５０＋Ｈ／ｍｌの溶液を調製した。ヒツジ赤血
球溶血液を浸透圧溶血法で調製した。Example 1 Carboxymethyl chitin (manufactured by Nanyo Kasei Co., Ltd.) was dissolved in water to make a 0.2% (W/V) solution. Dekis) Lan (Mw73,300 Sigma Chemical Company
St. Louis) was dissolved in Tris-HCl buffer (pH 7.4) to make a 6% (W/\・") solution.Egg yolk lecithin (manufactured by Asahi Kasei Corporation) was dissolved in chloromethane in a nitrogen stream. A solution with a concentration of 50+H/ml was prepared.Sheep red blood cell lysate was prepared by osmotic hemolysis.
上記ヒツジ溶血液１０ｍ１にレシチン溶液１（，１ｍ
ｌを加えて、マグネットスターラで１分間激しく攪拌し
Ｗ２Ｏ型のエマルジョンを調製した。このエマルジョン
を３１３ ｒｐｍで攪拌しなからすばやくカルボキシメ
チルキチン１（Ｎ）＋ｎｌｌこ力１１えＷ ／ Ｏ／Ｗ
型のエマルジョンを得た。攪拌を続けながら１０分後に
、更にカルボキシメチルキチン水溶液１００ｍ１を加え
、ジクロロメタンが完全に蒸発するまで攪拌を続行し、
ヒツン゛ヘモグロビン包接リポソーム（以下、単にＡＲ
ＢＣとい−う）を得た。この操作中、温度は室温に保っ
た。得られた水分散液を１２，０００ｇで１時間遠心分
離にかけ、Ａ ＲＢＣを捕集した。このＡＲＢＣを蒸留
水で洗浄後、トリス−塩酸緩衝液（ｐＨ７，４イオン強
度＝０゜１５４）に再分散させ、それぞれ粒子濃度が１
０゜２０ 、３０及び４０％（Ｖ／Ｖ）のＡＲＢｃ分散
液を調製した。また、同様にしてヒツジ赤血球（以下、
単に５ＲＢＣという）分散液（ｐＨ７、４）を調製した
。10ml of the above sheep blood lysate to 1ml of lecithin solution (1ml
1 was added and stirred vigorously for 1 minute using a magnetic stirrer to prepare a W2O type emulsion. This emulsion was stirred at 313 rpm and then quickly mixed with carboxymethyl chitin 1 (N) + nll power 11 w/o/w.
A mold emulsion was obtained. After 10 minutes while continuing stirring, 100 ml of carboxymethyl chitin aqueous solution was added and stirring was continued until dichloromethane was completely evaporated.
Human hemoglobin clathrate liposome (hereinafter simply AR
BC) was obtained. The temperature was kept at room temperature during this operation. The resulting aqueous dispersion was centrifuged at 12,000 g for 1 hour to collect ARBC. After washing this ARBC with distilled water, it was redispersed in Tris-HCl buffer (pH 7,4 ionic strength = 0°154), and the particle concentration was 1.
0°20, 30 and 40% (V/V) ARBc dispersions were prepared. Similarly, sheep red blood cells (hereinafter referred to as
A dispersion (simply referred to as 5RBC) (pH 7.4) was prepared.
ＡＲＢＣと５ＲＰＣの懸濁液の流動特性はコントロール
装置（レオフート３０．コントレイプズインダストリア
ルプロダク）Ｌｔｄ、スイス）を備えた集中円筒型回転
粘度計（ＬＳ−１００，同コントレイブス’Ｌｔｄ、）
で測定した。測定に使用した懸濁液量は１「ｒｌｌ と
し、温度は２５±０．５°Ｃ一定とした。The flow characteristics of the ARBC and 5RPC suspensions were measured using a concentrated cylindrical rotational viscometer (LS-100, Contraves Industrial Products Ltd., Switzerland) equipped with a control device (Leofut 30, Contraves Industrial Products Ltd., Switzerland).
It was measured with The amount of suspension used in the measurement was 1"rll, and the temperature was kept constant at 25±0.5°C.
得られたＡＲＢＣを走査電子顕微鏡（ｘ ５ 、０００
）により観察した結果、ＡＲＢＣは平均粒径３１０ｎ
ｍの完全な球形であることかわかった。従って、これら
の血球は毛細血管を自由に通過し得る。The obtained ARBCs were subjected to scanning electron microscopy (x 5,000
), the average particle size of ARBC was 310n.
It turns out that it is a perfect sphere of m. These blood cells can therefore freely pass through the capillaries.
第１図は、上記ＡＲＢＣ懸濁液の濃度を変えた場合の流
動特性を示す。また、第２図は、粒子濃度を変数とする
ＡＲＢＣと５ＲＢＣ懸濁液（ＩＩＨ７，４）の比粘度を
示す。５ＲＢＣＩＩ濁液の比粘度は粒子濃度と共に著し
く増大する。これに対して本発明のＡＲＢＣは実験範囲
では直線的に増加する。即ち、５ＲＢ（Ｊ濁液の比粘度
と粒子濃度との関係はブリンクマン（Ｂ ｒ ｉ ｎｋ
＋ｎａｎ ）の式とよく一致するが、ＡＲＢＣ懸濁液の
比粘度は約２．３の傾きを有するＥｉｎｓｔｅｉｎの式
にしたかう。FIG. 1 shows the flow characteristics when the concentration of the ARBC suspension is changed. Moreover, FIG. 2 shows the specific viscosity of ARBC and 5RBC suspensions (IIH7,4) with particle concentration as a variable. The specific viscosity of the 5RBCII suspension increases significantly with particle concentration. In contrast, the ARBC of the present invention increases linearly in the experimental range. That is, the relationship between the specific viscosity of the 5RB (J suspension) and the particle concentration is expressed by Brinkmann
+nan ), but the specific viscosity of the ARBC suspension is determined by Einstein's equation, which has a slope of about 2.3.
一方、血液と赤血球懸濁液の流動特性は力７ソン（Ｃａ
ｓｓｏｎ ）プロントに従うすることか゛知られている
。第３図は上記ＡＲＢＣｊｌ濁液のカッランプロットを
示し、ＡＲＢＣ懸濁液は直線性を示すことがわかる。On the other hand, the flow characteristics of blood and red blood cell suspensions are
sson) It is known to follow the prompt. FIG. 3 shows a Callan plot of the ARBCjl suspension, and it can be seen that the ARBC suspension exhibits linearity.
第４図は、６％（Ｗｌｏ）のデキストランを含むトリス
−塩酸緩衝溶液中におけるＡ ＲＢ Ｃ懸濁液の流動特
性を示す。ここで、デキストランは代用血しょう又は増
粘剤として使用されている。第１１図と第１図を比較す
ると、デキス）・ランを用いたＡＲＢＣｉｌｌ濁液の流
動抵抗はデキストランなしのＡＲＢＣ懸濁液よりも低ず
れ速度においてより高くなることか゛わかる。また、第
５図はデキストラン濃度を変数としたときのＡ ＲＢ
Ｃ及び５ＲＢＣの４０％（Ｖ／＼７）＠濁液の比粘度を
示す。ＡＲＢＣ懸濁液の比粘度は、Ｓ ＲＢ Ｃ懸濁液
の場合と同様に、デキストラン濃度の増加と共に減少す
る。FIG. 4 shows the flow properties of an A RBC suspension in a Tris-HCl buffer solution containing 6% (Wlo) dextran. Here, dextran is used as a plasma substitute or thickener. Comparing FIG. 11 with FIG. 1, it can be seen that the flow resistance of the ARBCill suspension with dextran is higher at low shear rates than the ARBCill suspension without dextran. In addition, Fig. 5 shows the A RB when the dextran concentration is used as a variable.
The specific viscosity of 40% (V/\7) @ suspension of C and 5RBC is shown. The specific viscosity of the ARBC suspension decreases with increasing dextran concentration, similar to that of the S RBC suspension.
デキス）・ラン濃度８％（Ｗ／Ｖ）では、比粘度かデキ
ストランなしのものと比べて２０％減少する。このこと
はＡ、 ＲＢ Ｃ及びＳ ＲＢ Ｃの流動特性はデキス
トランの存在によりかなり影響を受けることを示してい
る。At a concentration of 8% (W/V) dextran, the specific viscosity decreases by 20% compared to that without dextran. This shows that the flow properties of A, RB C and S RB C are significantly affected by the presence of dextran.
また、第６図は、ＡＲＢＣと５ＲＢＣの混合懸濁液の混
合比と比粘度との関係を示す。この混合懸濁液の総粒子
濃度は４０％一定とした。分散媒としてはデキストラン
を使用したものと使用しないものとかある。５ＲＢＣに
対するＡＲＢＣの混合比率を犬にすると、懸濁液の比粘
度はデキストラン濃度に関係なく減少する傾向を示す。Moreover, FIG. 6 shows the relationship between the mixing ratio and specific viscosity of a mixed suspension of ARBC and 5RBC. The total particle concentration of this mixed suspension was kept constant at 40%. Some use dextran and others do not use dextran as a dispersion medium. When the mixing ratio of ARBC to 5RBC is increased, the specific viscosity of the suspension tends to decrease regardless of the dextran concentration.
ＡＲＢＣの比率か減少すると、比粘度はＳ ＲＢ Ｃ懸
濁液（ｐＨ７、４，）に近づく。As the proportion of ARBC decreases, the specific viscosity approaches that of SRBC suspension (pH 7.4).
このように、本発明−（こよるＡＲＢＣの懸濁液（＋１
１−１７ 、４． ）の流動特性は擬似塑性であり、５
ＲＢ（］％濁液と同じ挙動を示す。ＡＲＢＣと５ＲＢＣ
懸濁液の比粘度は何れも分散媒中の粒子濃度と共に増加
する。しかしなから、Ａ ＲＢ Ｃ懸濁液についての比
粘度と粒子濃度との関係はＥｉｎｓＬｅｉｎの式によく
一致するのに対し、５ＲＢＣｊ％濁液ではＢｒｉｎｋｍ
ａｎの式に適合した１、これは、機械的強化剤として添
加したカルボキシメチルキチンかＡＲＢＣの表面にイオ
ン結合により吸着されているためと考えられる。Thus, according to the present invention, the suspension of ARBC (+1
1-17, 4. ) is pseudoplastic, and 5
Shows the same behavior as RB(]% suspension.ARBC and 5RBC
The specific viscosity of any suspension increases with the particle concentration in the dispersion medium. However, the relationship between specific viscosity and particle concentration for ARBC suspensions is in good agreement with EinsLein's equation, whereas for 5RBCj% suspensions, Brinkm
1, which conformed to the formula of an, is thought to be because carboxymethyl chitin, which was added as a mechanical reinforcement, was adsorbed on the surface of ARBC by ionic bonding.
一方、ＡＲＢＣとＳ ＲＢ Ｃの混合懸濁液（１＋１−
１７．４）の流動特性については、Ａ丁ぐＦ３ＣとＳ
Ｉり８０間の相互作用がＡ ＲＢ Ｃの混合比率を犬に
するにしたがって弱まることがわかる。そして、分散媒
中にデキストランを添加しても、その流動特性について
殆ど影響を与えない。従って、混合懸濁液中におけるＡ
ＲＢＣは、カルボキシメチルキチンの強い水利力によっ
て粒子−粒子間の相互作用を減少させると推論される。On the other hand, a mixed suspension of ARBC and SRB C (1+1-
Regarding the flow characteristics of 17.4),
It can be seen that the interaction between I and R80 weakens as the ARBC mixing ratio increases. Even if dextran is added to the dispersion medium, its fluidity properties are hardly affected. Therefore, A in the mixed suspension
It is inferred that RBC reduces particle-particle interactions due to the strong hydration of carboxymethyl chitin.
実施例２
２０ｍ１の２０％ヒトヘモグロビン液へ２０１１１１＜
７）レシチンのクロロホルム溶液を加え、磁気的攪拌器
で１分間激しく攪拌し、Ｖｖ’１０型のエマルジョンを
得−る。このエマルジョンに０．３％キトサンの水溶液
１００ ｍｌをすばやく加え４００ ｒｐ＋ｒで攪拌す
る。さらに、１０分後、２００＋ｎｌのカルボキシメチ
ルキトサンを加え、クロロホルムが蒸発するまで続ける
。なお、この一連の操作は室温で行う。得られたリポソ
ームは、ヒト赤血球と同じ酸素運搬能を有する。Example 2 To 20ml of 20% human hemoglobin solution 201111<
7) Add a chloroform solution of lecithin and stir vigorously for 1 minute using a magnetic stirrer to obtain a Vv'10 type emulsion. 100 ml of a 0.3% chitosan aqueous solution was quickly added to this emulsion and stirred at 400 rp+r. After another 10 minutes, add 200+ nl of carboxymethyl chitosan and continue until the chloroform has evaporated. Note that this series of operations is performed at room temperature. The resulting liposomes have the same oxygen carrying capacity as human red blood cells.
実施例３
３０＋ｏｇ／＋ｎレシチンと少量のコレステロールを含
むジクロロメクン溶液１部と４０Ｕ／ｍｌのインシュリ
ン水溶液１部とを乳化し、Ｗ／○型エマルノヨンとする
。次に、このエマルジョンを攪拌系にある０、０２％（
Ｗ／＼ｌ）カルボキシメチルキチン水溶液にすばやく加
え、Ｗ／○／Ｗ型エマルジョンとする。 その後、昇温
しでジクロロメタンを蒸散させ、インシュリン封入のリ
ポソームとする。Example 3 1 part of a dichloromecne solution containing 30+og/+n lecithin and a small amount of cholesterol and 1 part of a 40 U/ml insulin aqueous solution are emulsified to form a W/○ type emulsion. Next, this emulsion was placed in a stirring system at 0.02% (
W/\l) Quickly add to the carboxymethyl chitin aqueous solution to make a W/○/W type emulsion. Thereafter, dichloromethane is evaporated by raising the temperature to form insulin-encapsulated liposomes.
実施例４
５０ｍｇ／ｍｌレシチンベンゼンン容液２部と５０ｍｇ
／ ｎ＋ ｌのアルブミン水溶液１部を機械的に乳化し
、Ｗ１０型エマルン゛ヨンとする。このエマルジン゛ヨ
ンを攪拌系にある（）、１％（Ｗ／Ｖ）キトサン水溶液
に分散し、Ｗ１０／Ｗ型エマルノヨンとし、その後糸の
温度を上げてベンゼンを蒸散させ、アルブミン封入のリ
ポソームとした。Example 4 2 parts of 50mg/ml lecithin benzene solution and 50mg
1 part of an albumin aqueous solution of /n+l was mechanically emulsified to form a W10 type emulsion. This emulsion was dispersed in a 1% (W/V) chitosan aqueous solution in a stirring system to form a W10/W type emulsion, and then the temperature of the thread was raised to evaporate benzene to form albumin-encapsulated liposomes. .
実施例５
ブドウ糖を含むカルボキシメチルキチン化したリポソー
ムの調製
まず、０．０１から０．５％（Ｗ／＼・“）までのカル
ボキシメチルキチン水溶液を用意する。Example 5 Preparation of carboxymethyl chitinated liposome containing glucose First, an aqueous solution of carboxymethyl chitin ranging from 0.01 to 0.5% (W/\•") is prepared.
内封物のフド・ン糖溶液１部と５（、）ｍ８／ｍｌの卵
黄レシチンジクロルメタン溶液１部を混合上攪拌するこ
とで、Ｗ１０型エマルノヨンとする。A W10 type emulsion is prepared by mixing and stirring 1 part of the enclosed fudon sugar solution and 1 part of a 5 m8/ml egg yolk lecithin dichloromethane solution.
このエマルジョンを」二記の各カルボ゛キシメチルキチ
ン水溶液、２００部中にすばやく加え、攪拌してＷ１０
／Ｗ型エマルジョンとする。これまでの操作はすべて室
温で行なう。This emulsion was quickly added to 200 parts of each carboxymethyl chitin aqueous solution mentioned above, stirred and
/W type emulsion. All previous operations are performed at room temperature.
次に、系の温度を４０℃まで上昇させると、ジクロルメ
タンがエマルジョン中から蒸散する。この過程で、リポ
ソームを構成してし）るレシチンと、カルボキシメチル
キチンとの静電的な結合が生じる。さらに、ジクロルメ
タンか完全に蒸散すると、カルボキシメチル化したリポ
ソームをイυることかで゛きる。Next, when the temperature of the system is raised to 40° C., dichloromethane evaporates from the emulsion. During this process, electrostatic bonding occurs between lecithin, which constitutes the liposome, and carboxymethyl chitin. Furthermore, complete evaporation of dichloromethane can destroy carboxymethylated liposomes.
−に記の方法に従ってブト゛つ糖含人リポソームを調製
した場合の各カルボキシメチルキチン濃度に対するブド
ウ糖のリポソーム・＼の取込み率を下の表に示す。The table below shows the uptake rate of glucose into the liposomes for each carboxymethyl chitin concentration when the butyl sugar-containing liposomes were prepared according to the method described in -.
図面は本発明の一実施例を示し、第１図は本発明による
ＡＲＢＣ懸濁液の濃度を変えた場合の流動特性図、第２
図は粒子濃度を変数とする−１−記ＡＲＢＣとＳｌマＢ
Ｃ懸濁液（ｐ Ｊ−１７、４）の比粘度を示すグラフ、
第３図は上記Ａ ｉ（Ｂ Ｃｊ％濁液の力、ラン プロ
ットを示すグラフ、第・・１図は６％（Ｗｌｏ）のデキ
ストランを含む）リス−塩酸緩衝溶液中におけるＡ Ｒ
Ｂ Ｃ懸濁液の流動特性図、第１３図はデキストラン）
良度な変数としたときのＡ ＲＦ３ Ｃ及び５ＲＢＣの
４０％（Ｖ／Ｖ）懸濁液の比粘度を示すグラフ、第６図
はＡ ＲＢ ＣとＳｌマＢＣの混合懸濁液の混合比と比
粘度との関係を示すグラフである。
特許出願人 味の素株式会社
０ １ ２ ３ ４ ５
すり速度 ＋ ５ｅｃ−’
□ ＩＱ ２０ ３０ ４０粒子濃度
１％（ν／ｖ）
（ずつ速度）１／２＋ （ＳｅＣ−１）”’１２３４５
すり速度 ＋ ｓｅｃ＝
第５１
デキストラン濃度 １％（Ｗ／Ｖ）The drawings show one embodiment of the present invention, and FIG. 1 is a flow characteristic diagram when the concentration of the ARBC suspension according to the present invention is changed, and FIG.
The figure shows -1- ARBC and SlmaB with particle concentration as a variable.
A graph showing the specific viscosity of C suspension (p J-17, 4),
Figure 3 is a graph showing the force of the above A i (B Cj % suspension, run plot, Figure 1 contains 6% (Wlo) dextran) A R in a Lis-HCl buffer solution.
Flow characteristic diagram of B C suspension, Figure 13 is dextran)
A graph showing the specific viscosity of a 40% (V/V) suspension of A RF3 C and 5RBC when taken as a good variable. Figure 6 shows the mixing ratio of a mixed suspension of A RF3 C and Slma BC. It is a graph showing the relationship between and specific viscosity. Patent applicant Ajinomoto Co., Inc. 0 1 2 3 4 5 Slip speed + 5ec-' □ IQ 20 30 40 Particle concentration
1% (ν/v) (speed) 1/2+ (SeC-1)''12345 Slide speed + sec= 51st Dextran concentration 1% (W/V)
Claims (3)
骨格成分とする多糖類を混合又は結合させてなることを
特徴とする活性物質含有リポソーム。(1) An active substance-containing liposome comprising a bimolecular layer of phospholipid mixed with or bound to a polysaccharide whose main skeleton component is I)-glucosamine.
メチルキトサン、キトサンよりなる群から選ばれた１又
は２以上の混合物である特許請求の範囲第１項記載の活
性物質含有リポソーム。(2) The active substance-containing liposome according to claim 1, which is a polysaccharide or a mixture of one or more selected from the group consisting of carboxymethyl chitin, carboxymethyl chitosan, and chitosan.
ある特許請求の範囲第１項記載の活性物質含有リポソー
ム。(3) The active substance-containing liposome according to claim 1, wherein the polysaccharide is hyaluronic acid and/or a derivative thereof.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP58078747A JPS59210013A (en) | 1983-05-04 | 1983-05-04 | Active substance-containing liposome |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP58078747A JPS59210013A (en) | 1983-05-04 | 1983-05-04 | Active substance-containing liposome |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS59210013A true JPS59210013A (en) | 1984-11-28 |
| JPH0518807B2 JPH0518807B2 (en) | 1993-03-15 |
Family
ID=13670478
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP58078747A Granted JPS59210013A (en) | 1983-05-04 | 1983-05-04 | Active substance-containing liposome |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS59210013A (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS59216894A (en) * | 1983-05-23 | 1984-12-06 | Asai Gerumaniumu Kenkyusho:Kk | Germanium-containing liposome |
| EP0387252A1 (en) * | 1987-08-25 | 1990-09-19 | Macnaught Pty Ltd | Lubricant composition for rheumatism. |
| FR2667072A1 (en) * | 1990-09-24 | 1992-03-27 | Bioetica Sa | Ternary complex of chitosan, calcium ions and lipids, process of preparation and their applications |
| EP0498471A2 (en) * | 1986-12-23 | 1992-08-12 | The Liposome Company, Inc. | Liposomes comprising a guanidino aminoglycoside |
| US5401511A (en) * | 1991-02-14 | 1995-03-28 | Baxter International Inc. | Binding of protein and non-protein recognizing substances to liposomes |
| US5409704A (en) * | 1985-06-26 | 1995-04-25 | The Liposome Company, Inc. | Liposomes comprising aminoglycoside phosphates and methods of production and use |
| FR2761912A1 (en) * | 1997-04-14 | 1998-10-16 | Capsulis | PROCESS FOR ADHERING A PRODUCT TO A SURFACE |
| JP2005500991A (en) * | 2001-03-29 | 2005-01-13 | ザ スクリップス リサーチ インスチチュート | Formulations containing captured active ingredients and uses thereof |
| JP2005298407A (en) * | 2004-04-12 | 2005-10-27 | Masahiko Abe | Polycation-modified liposome and method for producing the same |
-
1983
- 1983-05-04 JP JP58078747A patent/JPS59210013A/en active Granted
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS59216894A (en) * | 1983-05-23 | 1984-12-06 | Asai Gerumaniumu Kenkyusho:Kk | Germanium-containing liposome |
| US5409704A (en) * | 1985-06-26 | 1995-04-25 | The Liposome Company, Inc. | Liposomes comprising aminoglycoside phosphates and methods of production and use |
| EP0498471A2 (en) * | 1986-12-23 | 1992-08-12 | The Liposome Company, Inc. | Liposomes comprising a guanidino aminoglycoside |
| EP0387252A1 (en) * | 1987-08-25 | 1990-09-19 | Macnaught Pty Ltd | Lubricant composition for rheumatism. |
| FR2667072A1 (en) * | 1990-09-24 | 1992-03-27 | Bioetica Sa | Ternary complex of chitosan, calcium ions and lipids, process of preparation and their applications |
| US5401511A (en) * | 1991-02-14 | 1995-03-28 | Baxter International Inc. | Binding of protein and non-protein recognizing substances to liposomes |
| FR2761912A1 (en) * | 1997-04-14 | 1998-10-16 | Capsulis | PROCESS FOR ADHERING A PRODUCT TO A SURFACE |
| WO1998046199A1 (en) * | 1997-04-14 | 1998-10-22 | Capsulis | Method for making a product adhere to a surface |
| US6277404B1 (en) | 1997-04-14 | 2001-08-21 | Capsulis | Method for making a product adhere to a surface |
| JP2005500991A (en) * | 2001-03-29 | 2005-01-13 | ザ スクリップス リサーチ インスチチュート | Formulations containing captured active ingredients and uses thereof |
| JP2005298407A (en) * | 2004-04-12 | 2005-10-27 | Masahiko Abe | Polycation-modified liposome and method for producing the same |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0518807B2 (en) | 1993-03-15 |
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