JP3329554B2 - Dried er - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to in vivo targeting as a drug delivery system or use as a sustained-release material, or use as an artificial oxygen carrier containing a hemoglobin (hereinafter, referred to as Hb) solution. The present invention relates to a dried product that enables long-term stable storage of many useful phospholipid vesicles (hereinafter referred to as vesicles), that is, dried phospholipid vesicles (hereinafter referred to as dried vesicles).

[0002]

2. Description of the Related Art Although many studies have been made on the application of a vesicle as a drug carrier, vesicles are unstable due to aggregation, fusion, or destruction by various stimuli. It is a general means to stabilize by making the additive or the polymerizable phospholipid component into a polymer.

With respect to the storage stability of the vesicle as a freeze-dried body, when a large amount of saccharides such as trehalose is mixed with the vesicle, hydrogen bonding between the sugar and the lipid polar portion occurs even after lyophilization, and the bilayer membrane is formed. It is reported that leakage of inclusions is suppressed when the structure is maintained and water is added to regenerate (LM.
Crowe et al, Biochm. Biophy
s. Acta 861, 131 (1986); Ta
Naka et al, Chem. Pharm. Bull
l. , 40, 1 (1992)), and a report on a freeze-dried powdering technique in which the same method was applied to an Hb-containing endoplasmic reticulum (hemosome) dispersion (AS Rudolf, Cr)
Yobiology, 25, 277 (1988)), the particle size of the endoplasmic reticulum after freeze-dried powder redispersion is maintained at 20% even when a large amount of trehalose of 300 mM is added, while maintaining the particle size of the endoplasmic reticulum before freezing. Hb leakage occurs,
In this case, the endoplasmic reticulum was not sufficiently stabilized.
Therefore, the dried vesicles obtained by the freeze-drying method have insufficient stability, and have a high sugar concentration when regenerated by adding water, and are practically difficult to use.

[0004] The endoplasmic reticulum containing polymerizable phospholipids as a main component
It has been reported that if bilayer is stabilized by polymerizing a polymerizable phospholipid after encapsulating b, there is no leakage of the inclusion even if freeze-thaw is repeated (E. Tsuchida,
Biomat. , Art. Cells & Immo
b. Biotech. 20, 337 (1992)), and there is also a report that the ER can be stored as a dry powder with the addition of a small amount of sugar (L. Wang et a).
1, Polym. Adv. Tech. 3,17. (19
92)). However, none of them is clear about the effects of polymerized phospholipids in living body administration.

[0005] On the other hand, it has been reported that a stable lyophilized product can be obtained by introducing a cholesterol derivative having a disaccharide in a hydrophilic portion onto the surface of the endoplasmic reticulum (RP Goodric).
het al, Biophys. J. 53, 121a
(1988)). In addition, α, α-
There is an invention in which trehalose trimicolate is introduced into vesicles and freeze-dried, and the vesicles have a reconstituting ability even when dispersed again in pure water (JP-A-1-754).
No. 21). However, all require introduction of a large amount of saccharides in order to obtain sufficient stability.

The present inventors have proposed an oligosaccharide lipid in which a long-chain alkyl is ester-bonded to an oligosaccharide having a clear structure (Japanese Patent Application Laid-Open No. 1-294701), and an ether bond at the terminal anomeric position of the oligosaccharide chain (Japanese Patent Application Laid-Open No. 294983 and Japanese Patent Application No. 4-206136) and the same amide bond (JP-A-5-317677 and Japanese Patent Application No. 4-19136).
No. 4 invention) and an oligoglycolipid having a clear ester-bonded structure (Japanese Patent Application No. 5-75016).
It has been disclosed that it can be used as an inhibitor of vesicle aggregation and fusion. This utilizes the effect of oligosaccharide chains extending from the endoplasmic reticulum surface to prevent access between the endoplasmic reticulum or between the endoplasmic reticulum and cells or proteins. He also disclosed that oligosaccharides have a better inhibitory effect. However, the use of the oligosaccharide lipid relates to stabilization of the dispersion of vesicles in an aqueous phase system, and there is no disclosure of matters relating to drying of vesicles.

As described above, those skilled in the art have already developed a technique for freeze-drying vesicles without polymerizing the membrane components. However, there is no disclosure of a technique for drying the ER without freezing. Therefore,
Those skilled in the art understand that adding sugars in the process of preparing freeze-dried endoplasmic reticulum results in the sugar chain being hydrogen-bonded to the lipid hydrophilic portion on the surface of the endoplasmic reticulum, and furthermore, the sugar chains also form hydrogen bonds. We believe that the lipid molecules forming the bilayer retain their configuration even after dehydration. However, a freeze-dried product obtained by adding a large amount of saccharide to a dispersion of vesicles has a problem such as an increase in osmotic pressure and viscosity during regeneration with addition of water due to a large amount of added sugar. Development of a method for efficiently coating the surface of the endoplasmic reticulum with a smaller amount of sugar has been awaited. Furthermore, the freeze-drying method is a method usually used as a dry powdering method of the vesicle dispersion, but when the saccharide is frozen from a dilute solution, the saccharide dissolved in the aqueous phase effectively covers the vesicle surface. There is a problem that the effect of adding sugar is low because it is dried without coating, and that the freezing operation itself destroys the endoplasmic reticulum. As a result, there is a great expectation for a method for efficiently coating the surface of the endoplasmic reticulum with a smaller amount of sugar, and for developing a stable dried endoplasmic reticulum that does not inhibit the function of the endoplasmic reticulum when applied to a living body.

[0008]

Accordingly, the present invention provides a dried vesicle which can be stored for a long period of time, and which can retain and express its original structure and function upon regeneration by adding water. Is a technical issue.

[0009]

Means for Solving the Problems The present invention relates to dried vesicles that can be obtained without the use of concentrated sugars to solve the above-mentioned problems. That is, the present invention relates to a dried vesicle characterized in that a glycolipid is introduced into a phospholipid vesicle and then dried under reduced pressure without freezing.

Hereinafter, the present invention will be described in detail. In the preparation of the dried endoplasmic reticulum of the present invention, the carbohydrate used for the glycolipid introduced into the phospholipid endoplasmic reticulum (hereinafter referred to as carbohydrate) has a structure in which a long-chain alkyl group or cholesterol is bonded to the carbohydrate. If it is a substance, it may be an extract from a living body or a chemical compound. Examples of the biologically-derived glycolipid include glyceroglycolipid and glycosphingolipid. Preferably, the oligoglycolipid in which the long-chain alkyl is ester-linked to the oligosaccharide having a clear structure, the structure in which the long-chain alkyl is ester-linked to the oligosaccharide chain terminal anomeric position, which the present inventors have already disclosed, as described above. Oligosaccharide lipids, oligoglycolipids with a clear structure in which the same alkyl is ether-linked to the terminal anomeric position of the oligosaccharide chain, or
Oligoglycolipids having a clear structure in which the alkyl is amide-bonded to the terminal anomeric position of the oligosaccharide chain are effective.

Next, glycolipids, preferably oligoglycolipids,
Is described as a component of the endoplasmic reticulum membrane component. Glycolipids and endoplasmic reticulum components such as phospholipids are dissolved in chloroform, ether, or a mixed solvent of these and methanol and dried under reduced pressure in an eggplant-shaped flask to form a thin film of the mixed lipid on the inner wall of the flask. Alternatively, the glycolipid and the endoplasmic reticulum component are dissolved in benzene, dioxane, or a mixed solvent thereof with methanol, and lyophilized to obtain a powder of the mixed lipid. The content of the glycolipid is 0.1 to 50 mol%, preferably 1 to 10 mol% based on the total lipid.
And An aqueous solution of Hb, a water-soluble drug or a fluorescent molecule is added to the mixed lipid thin film or powder, and the mixture is shaken or stirred. The dispersion obtained here can be further irradiated with ultrasonic waves, or can be passed through a filter having different pore sizes in multiple stages by an extrusion method to control the dispersion to a desired particle size. The concentrated glycolipid-introduced endoplasmic reticulum can be obtained by removing the unencapsulated substance in the outer aqueous phase using a gel filtration column or an ultracentrifuge, and then performing centrifugal concentration. Another preparation method is to add a glycolipid dispersion to a functional substance-encapsulated vesicle dispersion and allow the mixture to stand, thereby introducing the glycolipid onto the surface of the vesicle membrane facing the external aqueous phase. is there.

Next, a drying operation is performed. The glycolipid-introduced vesicle dispersion liquid is dried by evaporating water while gradually increasing the degree of vacuum at room temperature using, for example, a rotary evaporator. At this time, a desiccator or the like can be used instead of the rotary evaporator. In the drying operation, the drying time can be shortened by allowing a drying agent such as silica gel blue or diphosphorus pentoxide to coexist in the dryer. It is important that the degree of pressure reduction does not become lower than the saturated vapor pressure. A vacuum is formed when almost water evaporates, and dried for 1 hour or more to obtain dried vesicles.

The saccharide is added to the glycolipid-introduced vesicle dispersion in an amount of 0 to 300 mM, preferably 1 to 150 mM, so as to adjust the colloid osmotic pressure to 300 mOsm or less.
M may be added and dried under reduced pressure without performing a freezing operation. Here, the saccharide is a water-soluble saccharide, for example, glucose, maltose, galactose, sucrose and the like, as well as oligosaccharides such as trehalose and raffinose, dextran, dextrin, and polysaccharides such as pullulan. The degree is not particularly limited.

In the dried endoplasmic reticulum of the present invention, when the oligosaccharide lipid-introduced vesicle dispersion or the natural glycolipid-introduced vesicle dispersion is dehydrated, the intervening carbohydrates and / or sugars and the endoplasmic reticulum are removed. Since both are concentrated, the sugar unit is efficiently hydrogen-bonded to the surface of the endoplasmic reticulum, resulting in a more stabilized endoplasmic reticulum structure.

To regenerate the dried vesicles obtained by drying under reduced pressure of the present invention, pure water may be added thereto and shaken. The stability of the dried endoplasmic reticulum can be evaluated from the change in the particle size of the regenerated endoplasmic reticulum obtained by light scattering measurement and the measurement of the leakage of the inclusions. The amount of inclusions leaked is determined by subjecting the regenerated ER dispersion to gel filtration and incorporating the ratio of the amount of inclusions present in the ER fraction and the fraction of inclusions leaked from the ER, or the fluorescent substance whose concentration has been quenched. When this occurs, the contained fluorescent substance emits fluorescence when leaked, and this is detected and calculated by fluorescence measurement. The dried vesicles of the present invention have the same particle size after rehydration as before the drying operation, and the leakage rate can be suppressed to about 10% or less.

[0016]

The use of oligoglycolipids is an effective means for introducing sugar chains to the surface of the endoplasmic reticulum and covering the surface efficiently. Regarding the sugar chain length of the carbohydrate, an extremely short one such as a monosaccharide is unlikely to have a sufficient effect, and when it becomes a polymer, it induces aggregation of the endoplasmic reticulum. The oligoglycolipid in the present invention has about 2 to 30 sugar chains and can impart sufficient stability to the dried endoplasmic reticulum. In the method of drying under reduced pressure of the present invention, since the dehydration is performed gently, there is no destruction of the endoplasmic reticulum due to freezing such as freeze drying. In addition, even with a small amount of added saccharide, the saccharide efficiently acts on the surface of the endoplasmic reticulum, and stable dried endoplasmic reticulum can be obtained. The vesicles obtained by redispersing this in water have no change in particle size, and the inclusions are almost completely retained. Therefore, the vesicles used for each purpose can be stably stored not as a dispersion but as a solid.

Next, when a large amount of an artificial oxygen carrier or the like containing Hb in the endoplasmic reticulum is administered in a large amount to the body, the osmotic pressure of the solution is required to be the same as the physiological condition. The dried ER requires only a small amount of added sugars during drying, so it does not cause an increase in osmotic pressure and is directly redispersed in water, or is administered to the living body immediately after adding the required amount of sugar to the ER redispersion solution it can. Further, since the particle size does not increase after the redispersion, the residence time of the endoplasmic reticulum in the blood does not decrease, and since the inclusions are hard to leak, the transport efficiency does not decrease.

[0018]

The present invention will be described in more detail with reference to the following examples. Embodiment 1 FIG. 96 mg of hexadecylmaltopentaonamide (hereinafter referred to as MPCA) (8.6 m
ol%) was dissolved in 2 ml of methanol, and this was dissolved in dipalmitoylglycerophosphatidylcholine (hereinafter referred to as DPPC).
Cholesterol (hereinafter referred to as Chol.)
And a mixed lipid composed of palmitic acid (hereinafter referred to as PA) (molar ratio: DPPC / Chol. / PA / MPCA)
= 7/7/2 / 1.5) 500 ml of chloroform solution containing 20 ml. Using the mixed solution, a thin film is formed on the inner wall of the eggplant-shaped flask using a rotary evaporator. Here, 10 ml of a purified aqueous solution of Hb having a concentration of 40 g / dl and purified from stroma was added, and the particle diameter was 200 ± 34 nm (Particlel) by an extrusion method.
eAnalyser N4-SD: trade name, manufactured by Coulter Corporation) was obtained.

0.5 ml of the Hb-containing vesicle dispersion was placed in a desiccator, coexisted with 5 g of diphosphorus pentoxide, and allowed to stand under reduced pressure (20 mmHg, 4 ° C.). After 3 hours, the resultant was vacuum-dried at 0.05 mmHg for 1 hour at room temperature of 21 ° C. using a vacuum pump to obtain dried vesicles. Next, the dried endoplasmic reticulum
After adding pure water at 5 ° C. and re-dispersing to an initial concentration, the mixture was subjected to gel filtration (10 mmd × 100 mmh, Sepharose CL-4B: trade name, manufactured by Pharmacia). From the Hb concentrations of the Hb endoplasmic reticulum fraction and the Hb solution fraction (quantified by the cyano-methemoglobin method), the Hb leakage rate was calculated to be 10.3%. The particle size distribution was 240 ± 83 nm, and almost no change in the particle size was observed before and after drying.

Comparative Example 1, which will be described later, showing the results of Example 1,
The results are compared with those of Comparative Examples 2 and 3. Comparative Example 1 In the Hb-encapsulated vesicle dispersion of Example 1,
The same vacuum drying as in Example 1 was performed for a system containing no MPCA (oligoglycolipid). When pure water was added to the dried product and redispersed, the Hb leakage rate was 62.4%. Therefore, it can be said that the introduction of the oligosaccharide lipid into the endoplasmic reticulum in Example 1 made the dried endoplasmic reticulum more stable.

Comparative Example 2 The Hb-containing vesicle dispersion liquid
5 ml was freeze-dried (0.04 mmHg, -30 ° C, 9 hours) and re-dispersed in the same manner.
3%. Further, the Hb-containing vesicle dispersion liquid was subjected to ultracentrifugation (50,000 × g, 60 minutes) to remove the supernatant, and trehalose was added to the outer aqueous phase with the osmotic pressure of blood (300 mOs).
m) The Hb leakage rate of the freeze-dried sample when 300 mM was added was 21.1%. Therefore, it was confirmed that the dried endoplasmic reticulum according to the present invention was extremely stable as compared with the freeze-dried endoplasmic reticulum.

Comparative Example 3 A glycolipid in which dodecyl alcohol is ether-bonded to the anomeric position of α-D-glucopyranose (1-O-dodecyl-α-D-glucose (hereinafter referred to as “1-O-dodecyl-α-D-glucose”)
DG)) and a mixed lipid obtained by freeze-drying from a benzene solution (molar ratio: DPPC / Chol. /
Hb-containing endoplasmic reticulum containing sucrose in the external aqueous phase from 500 mg (PA / DG = 7/7/2 / 1.5) of 500 mg (using 8.6 mol% of DG based on the total lipid) according to the same method as in Comparative Example 2. Dispersion (lipid concentration 10 g / dl, average particle size 227 ±
78 nm) to obtain dried vesicles. The Hb leakage rate after redispersion of the dried vesicles was 45.2%, the average particle size was polydispersed at 260 ± 117 nm, and the turbidity was increased. Therefore, as a glycolipid to be introduced into the dried endoplasmic reticulum, a monosaccharide derivative was not suitable for imparting stability to the dried endoplasmic reticulum.

Embodiment 2 FIG. Using the same material as in Example 1, the Hb-containing vesicle dispersion obtained by the same method was subjected to ultracentrifugation (50,000 × g, 60 minutes), the supernatant was removed, and an aqueous sucrose solution was added. Disperse and total lipid concentration 5
g / dl and sucrose concentration of 146 mM. The H
b) 0.5 ml of the endoplasmic reticulum re-dispersed liquid
In the same manner as described above, 5 g of diphosphorus pentoxide was allowed to coexist, followed by standing under reduced pressure.
mmHg, 4 ° C). After 3 hours, the resultant was vacuum-dried at room temperature of 19 ° C. with a vacuum pump at 0.05 mmHg for 1 hour to obtain dried vesicles. Next, pure water was added to the dried vesicles at 25 ° C., re-dispersed to the initial concentration, and then subjected to gel filtration. The method was the same as in Example 1. Thus, the Hb endoplasmic reticulum fraction and the Hb solution fraction Hb
From the concentration, the Hb leakage rate was calculated to be 3.1%. The particle size distribution was 197 ± 42 nm, and almost no change in the particle size was observed before and after drying. The dried endoplasmic reticulum was sealed in an ampoule bottle under nitrogen and stored at room temperature. After 30 days, pure water was added and redispersed. Therefore, it became clear that the dried endoplasmic reticulum of the present invention can be stored for a long period of time.

Embodiment 3 FIG. Stearyl laminariheptaose (hereinafter referred to as SLH) in which a hydrophobic group is ether-bonded to a sugar chain terminal anomeric position as an oligoglycolipid (hereinafter referred to as SLH) (5.8 mol% based on the total lipid), and dimyristoyl glycerophosphatidylcholine (hereinafter referred to as DMPC) , Cho
l. Together with dipalmitoyl glycerophosphatidic acid (hereinafter referred to as DPPA) (molar ratio: DMPC / C
hol. / DPPA / SLH = 7/7/2/1) It was dissolved in a benzene / methanol mixed solvent and freeze-dried. 5 g (6) -carboxyfluorescein (hereinafter referred to as CF) aqueous solution (20 m
M, pH 7.4, trehalose 100 mM) 100 ml
At 4 ° C. using a magnetic stirrer.
Stir for 4 hours. The vesicle dispersion thus obtained was treated with a microfluidizer at 4 ° C. and 8000 psi for 10 minutes to obtain a vesicle dispersion having an average particle size of 100 ± 41 nm. After removing the CF in the outer aqueous phase, it was concentrated and the lipid concentration was 5 g
/ Dl, 50 mM of trehalose was added, and the mixture was dried under reduced pressure in the presence of diphosphorus pentoxide.
mHg, 25 ° C, 1 hour). When the dried endoplasmic reticulum was redispersed, the leakage rate of CF was 3.0%, and the average particle size was 102 ± 49 nm. In addition, 4
After storage at 7 ° C for 7 days, the re-dispersed endoplasmic reticulum
03 ± 54 nm, the leakage rate was 3.1%, which was extremely stable.

Embodiment 4 FIG. Purine hydrolyzate of corn starch was obtained by using liquid chromatography and keeping the temperature at 60 ° C. using an octadecyl-modified silica gel column (ODS-AQ: trade name, manufactured by YMC) as a stationary phase and pure water as a mobile phase. The reduced terminal of the obtained oligosaccharide having a polymerization degree of 25 was oxidized to a carboxylic acid. Oligoglycolipid in which the carboxylic acid of the oligosaccharide and 1,2-di-O-octadecyl-3-glycerol are ester-linked, that is, 1,
Dissolve 9.6 mg of 2-di-O-octadecyl-3-α (β) -eicosapentaonyl-rac-glyceride (3.0 mol% based on the total lipid) and 50 mg of DPPC powder in a mixed solvent of methanol / dioxane. And freeze-dried to obtain a dry powder. 2.5 ml of a phosphate buffer (50 mM, pH 7.4) containing 40 mg of calcein was added thereto, and the mixture was subjected to ultrasonic irradiation at 0 ° C. for 30 minutes in a nitrogen atmosphere to obtain an endoplasmic reticulum dispersion having an average particle size of 55 ± 5 nm. Prepared. At this time, it passed through a sterile filter having a pore size of 3 μm. Then, non-encapsulated calcein in the external aqueous phase was removed by gel filtration. Therefore, the vesicle dispersion obtained by adding 60 mM maltose was dried under reduced pressure in the presence of diphosphorus pentoxide, and then dried under vacuum (0.05 mmHg, 25 ° C., 11 hours) and redispersed. Was 3.2%, the average particle size was 58 ± 9 nm, and the dried endoplasmic reticulum obtained in this example was extremely stable.

Embodiment 5 FIG. Monosialoganglioside (G
M1: trade name, manufactured by Sigma Corporation (10.0 m with respect to total lipids)
ol%) and mixed lipid (molar ratio: DPPC / Chol. /
Dipalmitoyl phosphatidylglycerol = 5/4 /
1) was dissolved in a mixed solvent of methanol / chloroform, and a thin film was formed on the inner wall of the eggplant-shaped flask as in Example 1. This was made into a multilayer vesicle dispersion liquid by vortexing, and then freeze-dried to obtain a mixed lipid powder. This powder was washed with 10 mM Tris buffer (pH 7.4) containing 50 mM maltose and 70 mM calcein.
0 ml was added and stirred at room temperature for 6 hours. A dispersion of vesicles having an average particle size of 200 ± 10 nm was obtained by an extrusion method, followed by centrifugation (30,000 × g, 30 minutes)
Washing and concentration yielded a vesicle dispersion having an average particle size of 199 ± 22 nm with a lipid concentration of 6 g / dl. To this, maltose was added to a concentration of 100 mM, and dried vesicles were prepared in the same manner as in Example 1. The dried endoplasmic reticulum was sealed with nitrogen and stored at 4 ° C. After 30 days, when the dried endoplasmic reticulum was redispersed, the average particle size was 202 ± 25.
The leakage rate of nm and calcein was as low as 2.1%, and was stable.

Embodiment 6 FIG. The reducing terminal group of maltopentaose is oxidized to a carboxylic acid, and maltopentaose monocholesteryl ester (hereinafter referred to as MP-Chol.) Esterified with cholesteryl bromide as a glycolipid is introduced into the endoplasmic reticulum. 1 for lipids
0.5 mol% was used. 109 mg of the glycolipid was dissolved in 2 ml of methanol, and this was dissolved in DPPC, Chol. And PA mixed lipid (molar ratio: DPPC / Cho)
l. / PA / MP-Chol. = 7/5/2/2) 50
It was mixed with 20 ml of a chloroform solution containing 0 mg. Thereafter, in the same manner as in Example 1, a thin film was formed on the inner wall of the eggplant-shaped flask using the rotary evaporator for the mixed solution. To this was added 10 ml of a purified aqueous solution of Hb containing stroma at a concentration of 40 g / dl, and an Hb-containing vesicle dispersion having a particle size of 202 ± 36 nm was obtained by an extrusion method. This dispersion was subjected to ultracentrifugation (50,000 × g,
60 minutes), the supernatant was removed, an aqueous sucrose solution was added, and the mixture was redispersed to make the total lipid concentration 5 g / dl and the sucrose concentration 146 mM.

0.5 ml of the Hb-containing vesicle dispersion was placed in a desiccator, coexisted with 5 g of diphosphorus pentoxide, and allowed to stand under reduced pressure (20 mmHg, 4 ° C.). After 3 hours, the resultant was vacuum-dried at 0.05 mmHg for 1 hour at room temperature of 23 ° C. using a vacuum pump to obtain dried vesicles. Next, pure water was added to the dried vesicles at 25 ° C. and redispersed to an initial concentration, followed by gel filtration (10 mmd × 100 mmh, Sepharose CL-4B: trade name, manufactured by Pharmacia). The Hb leakage rate was calculated from the Hb concentration of the Hb endoplasmic reticulum fraction and the Hb solution fraction to be 3.2%.
The particle size distribution was 198 ± 37 nm, and almost no change in particle size was observed before and after drying. The dried endoplasmic reticulum was sealed in an ampoule bottle under nitrogen and stored at room temperature. After 30 days, pure water was added and redispersed. The particle size distribution was 201 ± 29 nm, and the leakage rate was 2.8%. Therefore, the dried endoplasmic reticulum of this example was also evaluated as being capable of long-term storage and having excellent stability.

[0029]

According to the present invention, dried vesicles obtained by introducing glycolipids into phospholipid vesicles and then drying them under reduced pressure without freezing have a change in particle size of the vesicles and leakage of inclusions. Since it is extremely small, it has become possible to store and store vesicles, which are widely expected as drug delivery systems and artificial oxygen carriers, as dry vesicles. Accordingly, the present invention has significant contributions to the art.

──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Shinji Takeoka 1-3-19-1 Nishiwaseda, Shinjuku-ku, Tokyo 201 (72) Inventor Hirosui Sakai 1-6-29, Sekichominami, Nerima-ku, Tokyo (72) Inventor Hidetoshi Tsuchida 2-10-10, Sekichominami, Nerima-ku, Tokyo Examiner Taizo Nakamura (56) References JP-A-64-75421 (JP, A) JP-A-5-178739 (JP, A) JP-A-5-317677 (JP, A) JP-A-2-184336 (JP, A) JP-A-62-500102 (JP, A) JP-A-4-500803 (JP, A) A) Special table Hei 2-502348 (JP, A) (58) Field surveyed (Int. Cl. ⁷ , DB name) A61K 9/127 B01J 13/02 A61K 38/16

Claims (9)

(57) [Claims] 1. A saccharide comprising an integer of 5 to 30 saccharide units
Changes in vesicle particle size distribution before and after rehydration, obtained by drying phospholipid vesicles containing oligoglycolipids with hydrophobic groups attached thereto under reduced pressure without lyophilization A stabilized, dried endoplasmic reticulum with no leakage of less than 10% after rehydration of the internally loaded drug without any. 2. The dried endoplasmic reticulum according to claim 1, wherein the oligosaccharide lipid has a hydrophobic group bonded to the anomeric position of aldose at the terminal. 3. A process according to claim 1 or 2 dried vesicles according binding of the hydrophobic group is an ester bond. 4. The method of claim 1 or 2 dried vesicles according binding of the hydrophobic group is an ether bond. 5. The bond of a hydrophobic group is an amide bond.
3. The dried endoplasmic reticulum according to 1 or 2 . 6. The hydrophobic group dried vesicle of claim 1, 2, 3, 4 also <br/> 5 wherein the hydrophobic group composed of an alkyl chain of 12 to 22 carbon atoms. 7. The hydrophobic group dried small in claim 1, 2, 3, 4 or 5, wherein the hydrophobic group composed of alkenyl chain having 12 to 22 carbon atoms with an unsaturated bond of 1 to 4 Endoplasmic reticulum. 8. The method according to claim 1, wherein the hydrophobic group is cholesterol.
The dried endoplasmic reticulum according to 1, 2, 3, 4 or 5 . 9. A saccharide composed of an integer of 5 to 30 saccharide units
Changes in vesicle particle size distribution before and after rehydration, characterized in that phospholipid vesicles containing oligoglycolipids with hydrophobic groups attached to them are dried under reduced pressure without lyophilization Leakage rate after rehydration of internally loaded drug
A method for producing stabilized dry vesicles of 0% or less.