JPH01180245A - Liposome - Google Patents
LiposomeInfo
- Publication number
- JPH01180245A JPH01180245A JP63003052A JP305288A JPH01180245A JP H01180245 A JPH01180245 A JP H01180245A JP 63003052 A JP63003052 A JP 63003052A JP 305288 A JP305288 A JP 305288A JP H01180245 A JPH01180245 A JP H01180245A
- Authority
- JP
- Japan
- Prior art keywords
- liposome
- acid
- red blood
- fatty acid
- artificial red
- 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
- 239000002502 liposome Substances 0.000 title claims abstract description 48
- 235000014113 dietary fatty acids Nutrition 0.000 claims abstract description 22
- 229930195729 fatty acid Natural products 0.000 claims abstract description 22
- 239000000194 fatty acid Substances 0.000 claims abstract description 22
- 150000004665 fatty acids Chemical class 0.000 claims abstract description 22
- 150000002632 lipids Chemical class 0.000 claims abstract description 22
- 102000001554 Hemoglobins Human genes 0.000 claims abstract description 13
- 108010054147 Hemoglobins Proteins 0.000 claims abstract description 13
- 239000000126 substance Substances 0.000 claims abstract description 10
- 125000004432 carbon atom Chemical group C* 0.000 claims description 6
- 150000004671 saturated fatty acids Chemical class 0.000 claims description 3
- 210000004369 blood Anatomy 0.000 abstract description 17
- 239000008280 blood Substances 0.000 abstract description 17
- 238000000034 method Methods 0.000 abstract description 9
- WQEPLUUGTLDZJY-UHFFFAOYSA-N pentadecanoic acid Chemical compound CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 abstract description 6
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 abstract description 4
- SZHOJFHSIKHZHA-UHFFFAOYSA-N tridecanoic acid Chemical compound CCCCCCCCCCCCC(O)=O SZHOJFHSIKHZHA-UHFFFAOYSA-N 0.000 abstract description 4
- 239000004094 surface-active agent Substances 0.000 abstract description 3
- 239000010409 thin film Substances 0.000 abstract description 3
- 235000021314 Palmitic acid Nutrition 0.000 abstract description 2
- TUNFSRHWOTWDNC-HKGQFRNVSA-N tetradecanoic acid Chemical compound CCCCCCCCCCCCC[14C](O)=O TUNFSRHWOTWDNC-HKGQFRNVSA-N 0.000 abstract description 2
- 230000002035 prolonged effect Effects 0.000 abstract 1
- 210000003743 erythrocyte Anatomy 0.000 description 27
- 230000002776 aggregation Effects 0.000 description 6
- 238000004220 aggregation Methods 0.000 description 6
- 238000005538 encapsulation Methods 0.000 description 6
- 239000008346 aqueous phase Substances 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 150000003904 phospholipids Chemical class 0.000 description 4
- JQWAHKMIYCERGA-UHFFFAOYSA-N (2-nonanoyloxy-3-octadeca-9,12-dienoyloxypropoxy)-[2-(trimethylazaniumyl)ethyl]phosphinate Chemical compound CCCCCCCCC(=O)OC(COP([O-])(=O)CC[N+](C)(C)C)COC(=O)CCCCCCCC=CCC=CCCCCC JQWAHKMIYCERGA-UHFFFAOYSA-N 0.000 description 3
- PORPENFLTBBHSG-MGBGTMOVSA-N 1,2-dihexadecanoyl-sn-glycerol-3-phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(O)=O)OC(=O)CCCCCCCCCCCCCCC PORPENFLTBBHSG-MGBGTMOVSA-N 0.000 description 3
- TWJNQYPJQDRXPH-UHFFFAOYSA-N 2-cyanobenzohydrazide Chemical compound NNC(=O)C1=CC=CC=C1C#N TWJNQYPJQDRXPH-UHFFFAOYSA-N 0.000 description 3
- BZTDTCNHAFUJOG-UHFFFAOYSA-N 6-carboxyfluorescein Chemical compound C12=CC=C(O)C=C2OC2=CC(O)=CC=C2C11OC(=O)C2=CC=C(C(=O)O)C=C21 BZTDTCNHAFUJOG-UHFFFAOYSA-N 0.000 description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 3
- 235000021360 Myristic acid Nutrition 0.000 description 3
- TUNFSRHWOTWDNC-UHFFFAOYSA-N Myristic acid Natural products CCCCCCCCCCCCCC(O)=O TUNFSRHWOTWDNC-UHFFFAOYSA-N 0.000 description 3
- 230000008034 disappearance Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000002504 physiological saline solution Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 235000021355 Stearic acid Nutrition 0.000 description 2
- 125000000217 alkyl group Chemical group 0.000 description 2
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- KEMQGTRYUADPNZ-UHFFFAOYSA-N heptadecanoic acid Chemical compound CCCCCCCCCCCCCCCCC(O)=O KEMQGTRYUADPNZ-UHFFFAOYSA-N 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 2
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000008117 stearic acid Substances 0.000 description 2
- 239000002253 acid Substances 0.000 description 1
- 230000004520 agglutination Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 235000021342 arachidonic acid Nutrition 0.000 description 1
- 229940114079 arachidonic acid Drugs 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- RNPXCFINMKSQPQ-UHFFFAOYSA-N dicetyl hydrogen phosphate Chemical compound CCCCCCCCCCCCCCCCOP(O)(=O)OCCCCCCCCCCCCCCCC RNPXCFINMKSQPQ-UHFFFAOYSA-N 0.000 description 1
- 229940093541 dicetylphosphate Drugs 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229940042880 natural phospholipid Drugs 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 235000003441 saturated fatty acids Nutrition 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
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
Landscapes
- 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)
- Manufacturing Of Micro-Capsules (AREA)
- Medicinal Preparation (AREA)
Abstract
Description
【発明の詳細な説明】
［産業上の利用分野］
本発明は、リポソームに関する。さらに詳しくは、本発
明はリポソームの脂質層に脂肪酸を含有するリポソーム
に関する。DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to liposomes. More specifically, the present invention relates to liposomes containing fatty acids in the lipid layer of the liposome.
本発明のリポソームは血中滞留時間が長く、またヘモグ
ロビンカプセル化効率やリポソームの凝集防止能も高い
ので、特に人工赤血球として有利に利用される。The liposome of the present invention has a long blood retention time, and also has high hemoglobin encapsulation efficiency and liposome aggregation prevention ability, so it is particularly advantageously used as an artificial red blood cell.
［従来の技術及び問題点］
リポソームは、人工赤血球にも応用されており、リポソ
ームの内部水相にヘモグロビン（Ｈｂ）をカプセル化し
たものが用いられている。この人工赤血球については、
リポソーム内へのＨｂのカプセル化効率を増大させるた
め、及びリポソームの凝集を抑制するために、負荷電リ
ン脂質、例えばホスファチジン酸やジセチルホスフェー
トなどをリポソームの脂質層中に含有させることが不可
欠とされている（Ｓｚｅｂｅｎｌ、 Ｊ、らによるｒＢ
ｌｏｃｈｅｓｌｓｔｒｙＪ ２４．２Ｂ２７ （１９８
５） ； Ｇａｂｅｒ。[Prior Art and Problems] Liposomes are also applied to artificial red blood cells, and hemoglobin (Hb) is encapsulated in the internal aqueous phase of liposomes. Regarding this artificial red blood cell,
In order to increase the encapsulation efficiency of Hb within liposomes and to suppress liposome aggregation, it is essential to include negatively charged phospholipids, such as phosphatidic acid and dicetyl phosphate, in the lipid layer of liposomes. (rB by Szebenl, J. et al.
locheslstryJ 24.2B27 (198
5); Gaber.
Ｂ、 Ｐ、らによるｒＰｒｏｇ、 Ｃｌ１ｎ、 Ｂｉｏ
ｌ、 Ｒｅｓ、、 Ｊ１６５、１７９ （１９８４）
）。rProg, Cl1n, Bio by B. P. et al.
l, Res, J165, 179 (1984)
).
しかし、一方人工赤血球の膜層にこの負荷電リン脂質を
含有させると、循環血中からの消失速度が速くなること
が知られている（Ｒ，Ｌ。However, it is known that when the membrane layer of artificial red blood cells contains this negatively charged phospholipid, the rate of disappearance from the circulating blood becomes faster (R,L).
ＪｕｌｌａｎｏらによるｒＢｌｏｃｈｅｍ、 Ｂｌｏｐ
ｈｙｓ、 Ｒｅｓ。rBlochem, Blop by Jullano et al.
hys, Res.
Ｃｏｍｍｕｎ、 Ｊ ［ｉ３．８５１ （１９７５）
） 、長い血中滞留時間が必須の要件とされる人工赤血
球にとって、血中からの消失速度が速いことは致命的な
問題であった。Commun, J [i3.851 (1975)
), the rapid rate of disappearance from the blood was a fatal problem for artificial red blood cells, which require a long residence time in the blood.
従って、血中滞留時間が長く、しかもカプセル化効率と
リポソームの凝集抑制能の高い人工赤血球の開発が望ま
れる。Therefore, it is desired to develop artificial red blood cells that have a long blood residence time, high encapsulation efficiency, and high ability to inhibit liposome aggregation.
［発明が解決しようとする問題点］
本発明の目的は、血中滞留時間が長く、しかもカプセル
化効率とリポソームの凝集抑制能の高いリポソーム特に
人工赤血球を提供することにある。[Problems to be Solved by the Invention] An object of the present invention is to provide a liposome, particularly an artificial red blood cell, which has a long residence time in blood and has high encapsulation efficiency and ability to inhibit liposome aggregation.
［問題点を解決するための手段］
上記の目的は、下記の構成を有する本発明によって達成
される。[Means for Solving the Problems] The above object is achieved by the present invention having the following configuration.
１）リポソームの脂質層に、負荷電付与物質として脂肪
酸を含有することを特徴とするリポソーム。1) A liposome characterized by containing a fatty acid as a negative charge imparting substance in the lipid layer of the liposome.
２）内部に物質を取り込んでなる１項記載のりボソーム
。2) The ribosome according to item 1, which incorporates a substance inside.
３）物質がヘモグロビンである２項記載のリポソーム。3) The liposome according to item 2, wherein the substance is hemoglobin.
４）脂肪酸の炭素数が、１２〜２２である１乃至３項の
いずれか１項に記載のリポソーム。4) The liposome according to any one of items 1 to 3, wherein the fatty acid has 12 to 22 carbon atoms.
５）脂肪酸が、直鎖状の飽和脂肪酸である１乃至４項の
いずれか１項に記載のリポソーム。5) The liposome according to any one of items 1 to 4, wherein the fatty acid is a linear saturated fatty acid.
リポソームは、脂質人工膜の一種であって脂質の２分子
膜層よりなる内部に水相を含んだ閉鎖小胞体である。本
発明のリポソームは、特に内水相にヘモグロビンを含有
する人工赤血球用のリポソームである。A liposome is a type of artificial lipid membrane, and is a closed endoplasmic reticulum containing an aqueous phase inside of a bimolecular lipid membrane layer. The liposome of the present invention is particularly a liposome for artificial red blood cells containing hemoglobin in the internal aqueous phase.
本発明において使用される脂肪酸は、脂溶性であって、
しかも他のリポソーム形成脂質とともにリポソームを形
成し得るものであれば特に限定はない。The fatty acids used in the present invention are fat-soluble,
Moreover, there is no particular limitation as long as it can form liposomes together with other liposome-forming lipids.
ただし炭素数が１２未満の脂肪酸は、水溶性あるいは親
水性が高く、またアルキル鎖の運動性も高いためリポソ
ームを不安定にしやすいので、炭素数が１２以上である
ことが望ましい。However, fatty acids with less than 12 carbon atoms tend to destabilize liposomes because they are highly water-soluble or hydrophilic and have high alkyl chain mobility, so it is desirable that the number of carbon atoms is 12 or more.
また、脂肪酸のアルキル基に側鎖がある場合には、リポ
ソーム脂質層の配列を乱しやすく、さらに、脂肪酸が不
飽和脂肪酸である場合には、毒性の高い過酸化物を生成
する可能性があるので、直鎖状の飽和脂肪酸が好ましく
用いられる。In addition, if the alkyl group of the fatty acid has a side chain, it tends to disturb the arrangement of the liposome lipid layer, and if the fatty acid is an unsaturated fatty acid, there is a possibility of producing highly toxic peroxides. Therefore, linear saturated fatty acids are preferably used.
なお、炭素数が２２を越えると、溶解性が低下、また希
少なために高価となるので、炭素数は２２以下が望まし
い。If the number of carbon atoms exceeds 22, the solubility decreases and it becomes expensive due to its scarcity, so it is desirable that the number of carbon atoms is 22 or less.
このような脂肪酸の例としては、トリデシル酸、ミリス
チン酸、ペンタデシル酸、パルミチン酸、ヘプタデシル
酸、ステアリン酸、アラキドン酸、トコサン酸等があげ
られる。Examples of such fatty acids include tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, heptadecylic acid, stearic acid, arachidonic acid, tocosanoic acid, and the like.
本発明において使用されるリポソームを形成する脂質は
、天然のリン脂質、合成の脂質類縁体等、リポソーム（
またはベシクル）を形成し得るものであれば特に限定さ
れるものではないが、卵黄レシチンまたは水素添加卵黄
レシチンとコレステロールとの混合物が好ましい。The liposome-forming lipids used in the present invention include natural phospholipids, synthetic lipid analogs, etc.
Although there are no particular limitations on the material as long as it can form vesicles), egg yolk lecithin or a mixture of hydrogenated egg yolk lecithin and cholesterol is preferred.
本発明のリポソームは、リポソーム形成脂質と脂肪酸の
混合物を常法によってリポソーム化することによって得
られる。人工赤血球を製造する場合は、赤血球膜が除去
されたヘモグロビン溶液をリポソーム化すればよい。リ
ポソーム形成脂質に対する脂肪酸のモル比は１％乃至２
０％（重量比でロ、゛３％乃至１３％）であり、５％乃
至１５％（重量比で１，５％乃至１０％）が望ましい。The liposome of the present invention can be obtained by forming a mixture of a liposome-forming lipid and a fatty acid into liposomes by a conventional method. When producing artificial red blood cells, a hemoglobin solution from which red blood cell membranes have been removed may be made into liposomes. The molar ratio of fatty acids to liposome-forming lipids is 1% to 2.
0% (3% to 13% by weight), preferably 5% to 15% (1.5% to 10% by weight).
人工赤血球中の脂肪酸の含有量は、上記範囲未満である
場合には、脂肪酸によるヘモグロビンのカプセル化効率
増大効果や人工赤血球の凝集防止効果が発現されにくい
ので望ましくなく、また、上記範囲を越える場合には、
脂肪酸の界面活性作用によってリポソームが不安定とな
り易いので望ましくない。If the content of fatty acids in the artificial red blood cells is less than the above range, it is undesirable because the effect of increasing hemoglobin encapsulation efficiency and the effect of preventing the aggregation of artificial red blood cells by fatty acids is difficult to be expressed, and if it exceeds the above range, it is undesirable. for,
This is undesirable because liposomes tend to become unstable due to the surfactant action of fatty acids.
リポソーム化は、常法に従って行なうことが可能であり
、例えば薄膜法、界面活性剤除去法、逆相法、フレンチ
プレス法等いづれの方法を用いてもよい。The formation of liposomes can be carried out according to conventional methods, such as a thin film method, a surfactant removal method, a reverse phase method, a French press method, and the like.
以下本発明を実施例とそれに対する比較例により詳しく
説明する。The present invention will be explained in detail below using examples and comparative examples.
実施例 １
水素添加卵黄レシチン２９．０ｇ （１９ａｍｏｌ）、
コレスチロール１４．５ｇ （３９ｍｍｏｌ）及びミリ
スチン酸１．２ｇ（５，３ｍ５ｏｌ）をジクロロメタン
中に均一に溶解した後、エバポレーションにより溶媒を
除去した。この混合脂質薄膜に蛍光色素カルボキシフル
オレセイン（ＣＦ）を２０ａ＋Ｈの濃度で含有する４０
％ヘモグロビン水溶液（１）Ｈ７，４） ３５０ｍ１を
加えて振とうし、脂質を分散させた後、２５０ｋｇ／ｃ
−の圧力でフレンチプレス処理を５回くり返した。得ら
れたフレンチプレス処理液を生理食塩水により１０倍に
希釈して遠心分離処理（１７，０ＯＯｒ、ｐ、ａ＋、３
０分）し、沈澱リポソームをさらに生理食塩水により遠
心洗浄を２回くり返して、カプセル化されなかったヘモ
グロビンと蛍光色素を除去した。得られた沈澱リポソー
ムを生理食塩水中に懸濁させ、孔径０．４５ｍのフィル
ターをン濾過後Ｈｂ濃度が２．５％になるように濃度を
調製してＨｂとＣＦを内水相に保持し、脂質層に脂肪酸
を含有した人工赤血球の懸濁液を得た。この懸濁液中の
全脂質濃度は８８．０ｍｇ／ｍｌであった。また、この
懸濁液中の人工赤血球には全く凝集は認められなかった
。Example 1 Hydrogenated egg yolk lecithin 29.0g (19amol),
After uniformly dissolving 14.5 g (39 mmol) of cholestylol and 1.2 g (5.3 m5 ol) of myristic acid in dichloromethane, the solvent was removed by evaporation. 40 containing the fluorescent dye carboxyfluorescein (CF) at a concentration of 20a+H in this mixed lipid thin film.
% hemoglobin aqueous solution (1) H7, 4) was added and shaken to disperse the lipids, then 250 kg/c
The French press process was repeated 5 times at a pressure of -. The obtained French press treatment solution was diluted 10 times with physiological saline and centrifuged (17,0OOr, p, a+, 3
0 minutes), and the precipitated liposomes were further centrifuged and washed twice with physiological saline to remove unencapsulated hemoglobin and fluorescent dye. The precipitated liposomes obtained were suspended in physiological saline, filtered through a filter with a pore size of 0.45 m, and the concentration was adjusted so that the Hb concentration was 2.5% to retain Hb and CF in the internal aqueous phase. A suspension of artificial red blood cells containing fatty acids in the lipid layer was obtained. The total lipid concentration in this suspension was 88.0 mg/ml. Furthermore, no agglutination was observed in the artificial red blood cells in this suspension.
この人工赤血球懸濁液を脂質量で８７０ｍｇ／ｋｇ体重
の投与量として、体重２００ｇのラットに静脈注射した
後、経時的に０．２ｍｌずつ採血した。各０．２ｍｌの
血液に蒸留水１．８ｍｌ及びイソプロピルアルコール２
ｍｌを加え、赤血球及びリポソームを破壊してヘモグロ
ビンを変性させた。遠心分離によって変性ヘモグロビン
沈澱分を除去して、上清中の蛍光量を測定することによ
り人工赤血球の血中消失過程を追跡したところ、第１図
の丸プロットの結果を得た。This artificial red blood cell suspension was intravenously injected into a rat weighing 200 g at a dose of 870 mg lipid/kg body weight, and then 0.2 ml of blood was collected over time. For each 0.2 ml of blood, 1.8 ml of distilled water and 2 ml of isopropyl alcohol.
ml was added to destroy red blood cells and liposomes and denature hemoglobin. The denatured hemoglobin precipitate was removed by centrifugation, and the amount of fluorescence in the supernatant was measured to track the disappearance process of the artificial red blood cells from the blood, and the results shown in the circle plot in FIG. 1 were obtained.
実施例 ２
実施例１のミリスチン酸の代りにステアリン酸５Ｊｍｍ
ｏｌを使用する以外は実施例１と全く同様の操作を行な
ったところ、実施例１と同様の結果を得た。Example 2 Stearic acid 5Jmm instead of myristic acid in Example 1
The same operation as in Example 1 was performed except that ol was used, and the same results as in Example 1 were obtained.
比較例 １
実施例１のミリスチン酸の代りにジパルミトイルホスフ
ァチジン酸５．３ｍｍｏｌを使用する以外は実施例１と
全く同様の操作を行なったところ、Ｈｂ濃度２．５％で
全脂質濃度３９．４ｍｇ／ｍｌの人工赤血球懸濁液を得
た。これについて実施例１と同様の動物実験を行なった
ところ、第１図の三角プロットの結果を得た。Comparative Example 1 The same operation as in Example 1 was performed except that 5.3 mmol of dipalmitoyl phosphatidic acid was used instead of myristic acid in Example 1. As a result, the total lipid concentration was 39.4 mg at an Hb concentration of 2.5%. /ml artificial red blood cell suspension was obtained. Regarding this, an animal experiment similar to that in Example 1 was conducted, and the triangular plot shown in FIG. 1 was obtained.
第１図から明らかなように、本発明のリポソームは８時
間で５０％以上、７２時間で１０％以上の血中残存率を
示すのに対して、従来リポソームは８時間で１％以下、
７２時間では０．１％の血中残存率である。As is clear from FIG. 1, the liposome of the present invention shows a residual rate in blood of 50% or more after 8 hours and 10% or more after 72 hours, whereas the conventional liposome shows a residual rate of 1% or more after 8 hours.
The residual rate in blood is 0.1% after 72 hours.
［発明の効果］
本発明によればリポソームの脂質層に脂肪酸を含有させ
ることによって循環血中での滞留時間が延長されたリポ
ソーム特に人工赤血球が提供される。[Effects of the Invention] According to the present invention, a liposome, particularly an artificial red blood cell, whose residence time in circulating blood is extended by containing a fatty acid in the lipid layer of the liposome is provided.
本発明の人工赤血球は、リポソームの脂質層に負荷電物
質として脂肪酸を用いることによって、従来の負荷電リ
ン脂質を用いる人工赤血球よりも循環血中における滞留
時間が著しく長い。By using a fatty acid as a negatively charged substance in the lipid layer of the liposome, the artificial red blood cell of the present invention has a significantly longer residence time in circulating blood than artificial red blood cells using conventional negatively charged phospholipids.
従って、特に長い血中滞留時間が不可欠である人工赤血
球として、有用性が高い。Therefore, it is particularly useful as an artificial red blood cell that requires a long residence time in the blood.
さらに本発明の人工赤血球は、従来の負荷電リン脂質を
用いた人工赤血球と同様またはそれ以上のヘモグロビン
カプセル化効率とインビトロでの凝集防止能も有する。Furthermore, the artificial red blood cells of the present invention also have hemoglobin encapsulation efficiency and in vitro aggregation prevention ability that are similar to or higher than artificial red blood cells using conventional negatively charged phospholipids.
第１図は、人工赤血球の血中残存率の経時変化を示すグ
ラフであり、−〇−は本発明の脂肪酸含有人工赤血球の
場合を、−Δ−は従来のホスファチジン酸含有人工赤血
球の場合を示すものである。FIG. 1 is a graph showing changes over time in the residual rate of artificial red blood cells in blood, where -〇- indicates the case of the fatty acid-containing artificial red blood cell of the present invention, and -Δ- indicates the case of the conventional artificial red blood cell containing phosphatidic acid. It shows.
Claims (1)
酸を含有することを特徴とするリポソーム。 ２）内部に物質を取り込んでなる請求項１記載のリポソ
ーム。 ３）物質がヘモグロビンである請求項２記載のリポソー
ム。 ４）脂肪酸の炭素数が、１２〜２２である請求項１乃至
請求項３のいずれか１項に記載のリポソーム。 ５）脂肪酸が、直鎖状の飽和脂肪酸である請求項１乃至
請求項４のいずれか１項に記載のリポソーム。[Scope of Claims] 1) A liposome characterized in that the lipid layer of the liposome contains a fatty acid as a negatively charged substance. 2) The liposome according to claim 1, which contains a substance therein. 3) The liposome according to claim 2, wherein the substance is hemoglobin. 4) The liposome according to any one of claims 1 to 3, wherein the fatty acid has 12 to 22 carbon atoms. 5) The liposome according to any one of claims 1 to 4, wherein the fatty acid is a linear saturated fatty acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63003052A JPH01180245A (en) | 1988-01-12 | 1988-01-12 | Liposome |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63003052A JPH01180245A (en) | 1988-01-12 | 1988-01-12 | Liposome |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH01180245A true JPH01180245A (en) | 1989-07-18 |
Family
ID=11546549
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63003052A Pending JPH01180245A (en) | 1988-01-12 | 1988-01-12 | Liposome |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH01180245A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0395382A2 (en) * | 1989-04-27 | 1990-10-31 | Nippon Oil And Fats Company, Limited | Macromolecular endoplasmic reticulum |
| JP2008517023A (en) * | 2004-10-19 | 2008-05-22 | マックス−プランク−ゲゼルシャフト・ツア・フェルデルング・デア・ヴィッセンシャフテン・エー・ファオ | Formulations using novel negative charge carriers containing alkylphosphocholines |
| CN104825395A (en) * | 2015-04-22 | 2015-08-12 | 合肥华方医药科技有限公司 | Total bufadienolide nonionic surfactant vesicles and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS52151718A (en) * | 1976-06-10 | 1977-12-16 | Univ Illinois | Production of capsulated hemoglobin |
| JPS60155109A (en) * | 1984-01-23 | 1985-08-15 | Terumo Corp | Liposome pharmaceutical |
| JPS61267509A (en) * | 1985-05-21 | 1986-11-27 | Junzo Sunamoto | Production of liposome having improved holding efficiency of glycoprotein in membrane |
-
1988
- 1988-01-12 JP JP63003052A patent/JPH01180245A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS52151718A (en) * | 1976-06-10 | 1977-12-16 | Univ Illinois | Production of capsulated hemoglobin |
| JPS60155109A (en) * | 1984-01-23 | 1985-08-15 | Terumo Corp | Liposome pharmaceutical |
| JPS61267509A (en) * | 1985-05-21 | 1986-11-27 | Junzo Sunamoto | Production of liposome having improved holding efficiency of glycoprotein in membrane |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0395382A2 (en) * | 1989-04-27 | 1990-10-31 | Nippon Oil And Fats Company, Limited | Macromolecular endoplasmic reticulum |
| JP2008517023A (en) * | 2004-10-19 | 2008-05-22 | マックス−プランク−ゲゼルシャフト・ツア・フェルデルング・デア・ヴィッセンシャフテン・エー・ファオ | Formulations using novel negative charge carriers containing alkylphosphocholines |
| KR101311980B1 (en) * | 2004-10-19 | 2013-09-26 | 막스-플랑크-게젤샤프트 츄어 푀르더룽 데어 비쎈샤프텐 에.파우. | Formulations containing alkylphosphocholines using novel negative charge carriers |
| US8828972B2 (en) | 2004-10-19 | 2014-09-09 | Max-Planck-Gesellschaft Zur Foerderung Der Wissenschaften E.V. | Formulations containing alkylphosphocholines using novel negative charge carriers |
| CN104825395A (en) * | 2015-04-22 | 2015-08-12 | 合肥华方医药科技有限公司 | Total bufadienolide nonionic surfactant vesicles and preparation method thereof |
| CN104825395B (en) * | 2015-04-22 | 2018-02-27 | 合肥华方医药科技有限公司 | A kind of total toadpoison lactone nonionic surfactant vesicle and preparation method thereof |
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