JPS6327440A - Glucosylated branched cyclodextrin-containing composition - Google Patents

Abstract

PURPOSE:A drug composition having improved water solubility of a water- insoluble or sightly water-soluble pharmacologically active substance, by incorporating the pharmacologically active substance with a glucosylated branched cyclodextrin. CONSTITUTION:(A) A water-insoluble or slightly water-soluble pharmacologically active substance such as steroid hormone agent, cardiac, neutral nervous system depressant, antibiotic or fat-soluble vitamin, etc., is incorporated with (B) a glucosylated branched cyclodextrin preferably a compound wherein 1-3 glucoses are bonded by alpha,1 6 bond to alpha-, beta- or gamma-cyclodextrin through oxygen atom of the cyclodextrin. The component B is premixed with 0.3-2.0pts.wt. water, blended with inclusion mol equivalent (usually equimolar) of the component A for 0.5 - several hours while stirring and powdered by a usually applicable drying method to give the aimed drug composition (inclusion compound).

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a pharmaceutical composition with improved water solubility, comprising a water-insoluble or poorly water-soluble pharmacologically active compound and a glucosylated branched cyclodextrin.

BACKGROUND OF THE INVENTION It is known that for poorly water-soluble drugs, the rate of dissolution is the rate-limiting step for gastrointestinal absorption. Therefore, if it is possible to increase the solubility of those drugs, it is possible to improve the bioavailability of the drugs. Natural α is used to improve the solubility of conventionally water-insoluble or poorly water-soluble drugs.

β or Fir-cyclodextrin has been used [Kaneto Kamikama, Pharmaceutical Journal 101(10), 857-873
(1981)).

Among cyclodextrins, β-cyclodextrin (
β-CD (hereinafter abbreviated as β-CD) has a wide range of inclusion targets and is highly effective, but the solubility of β-CD itself is 1 at room temperature.
．． The solubility of the clathrate compound, which was quite low at 85% (W/v), was not necessarily satisfactory. Therefore, various derivatives have been created with the aim of increasing the solubility of β-CD, and their performance as solubilizers has been investigated.
-CD.

β-CD? However, these chemical derivatives, especially dimethyl-β-CD
Alternatively, trimethyl-β-CD is highly hemolytic and has a practical problem in terms of safety. [Odai et al. 6th
Abstracts of symposium on interaction between regenerative biomembranes and drugs, p.
50 (1983)).

The present inventors conducted an intensive search for cyclodextrins (abbreviated as CD) that are highly effective in solubilizing various water-insoluble or poorly water-soluble drugs and have low toxicity such as hemolytic properties.
Branched CD, which is a reaction product of starch with a CD-producing enzyme, has high solubility in water, and has been found to have a remarkable effect on increasing the solubility of water-insoluble or poorly water-soluble drugs, and is also highly toxic due to hemolytic properties. The present invention was completed by discovering that the

Therefore, the present invention provides a composition with improved water solubility in which a water-insoluble or poorly water-soluble pharmacologically active substance and a glucosylated branched cyclodextrin coexist.

In the present invention, water-insoluble or poorly water-soluble pharmacologically active substances include fat-soluble vitamins that are hardly soluble in water, such as vitamin E + K1 aK, , 0
3 or steroid hormones with extremely low solubility in water, non-steroidal anti-inflammatory drugs, central nervous system depressants such as phenoparbital and nitrazenome, cardiac glycosides such as digitoxin and dibuxin, or griseofulvin. Water-insoluble antibiotics such as

The glucosylated branched cyclotextrin used in the present invention is α-1β- or γ-CD whose constituent glucose is composed of 6, 7 or 8 constituent glucose. Glucose is bonded through α, 1→6 bonds via oxygen at position. Normally, it is possible to make the branched glucose chain length 1 glucose by applying glucoamylase to an aqueous solution of a mixture of various branched cyclodextrins (for example, Starch Science 30°23
pp. 1-239 (1983).

Among these, those in which α- or β-CD has one or two glucose bonds, α, 1→6, are preferably used.

Glucosylated branched cyclodextrin can be produced by isolation from the usual CD production reaction solution by various separation methods, such as solvent precipitation, column methods using various resin carriers, ultrafiltration, or reverse osmosis. It is possible. C
The D production reaction solution is a mixture of starch or its hydrolyzate with cyclodextrin glucanotransferase (hereinafter referred to as CGT).
E, C, 2, 4, 1, 19), abbreviated as aae, is 50
It is obtained by reacting at ~70°C and pH 5-10 for 18-72 hours.

Glycosylated branched cyclodextrins are synthesized by enzymes that cleave α, 1→6 bonds, such as isoamylases (E, C, 3,
It is also possible to produce using the reverse reaction of 2, 1, 68).

The inclusion method of the cosylated branched cyclodextrin and the water-insoluble or poorly water-soluble pharmacologically active substance can be carried out in the same manner as in the case of conventional CD and dust compounds.

That is, water is added to glucosylated branched cyclotextrin by 0.3
Add ~2.0 parts, knead in advance, and add clathrate molar equivalents (
Add a water-insoluble or poorly water-soluble pharmacologically active substance (dust compound) (usually equimolar), stir for 0.5 to several hours, and then dry by a commonly used drying method such as freeze drying, spray drying, ventilation drying, etc. By powdering, the desired clathrate compound can be obtained.

Effects of the Invention The present invention will be explained in more detail with experimental examples and examples.

Suspend 10 ok of potato starch in 1.0 kl of water, add Neosvitase (Nagao Sangyo), and stir at 80°C for 10 oz.
After liquefaction by heating for a minute, the mixture was sterilized at 120°C for 10 minutes. After cooling the liquefied liquid to 60°C, Bacillus□hbensi
s (FERM P-1990) CGTa s s
Added 10 u/g-f 5tarch amount, 60℃,
The CD production reaction was carried out at pH 7.6 for 40 hours. After the reaction, the enzyme was deactivated by treatment at 100°C for 10 minutes, the reaction solution was condensed to 2501 in a concentrator, and cooled at 10°C.
After one day of standing, the crystallized β-CD was distributed door to door. Add 0.01 glucoamylase (Ohno Pharmaceutical, GNL-2) to solution F (
Wt/lJ), reacted at 40°C for 5 hours, and CD
In addition to converting other dextrins to glucose, the side chains of the glycomolized branched CDO long chains were shortened to glucose units. Glucose was removed and concentrated from the glucoamylase-treated liquid using a reverse osmosis filter (ULVAC Service, RO-B: module: As-215) to obtain a concentrated liquid 251. Concentrate 251 to 501
After removing the precipitated β-CD fraction, 150 μl of acetone was added to remove the glucosylated branch C.
D crude fraction 2,5) C9 was obtained. Five crude fractions of the branched CD were subjected to Toyopearl HW-408 (Toyo Soda Kogyo Co., Ltd.) 2μ chromatography K, and 6-0-α-D-glucosyl α-CD (hereinafter abbreviated as Gα-CD)2. 05! y
-16-〇-α-D-glucosyl-β-〇〇 (hereinafter G
β-CD) 0.48/- and 6.6#-G0
-α-D-glufucyru β-CD (hereinafter referred to as 2Gβ-CD
) 0.97i! I got -.

The structure of the obtained branched CD was determined by its behavior on various HPLC columns, chromatography, 7-fragment analysis of mass quictor, analysis of methylated branched CD and its partially melted product,
Measurement items such as FAB-Mass 4 vectors, 6-〇-α-D-glucosyl-α-CD, 6-0-α-D-glucyl-β-CD and 6.6'-di0-α-D −
It was confirmed that it was glucosyl-β-CD. Table 1 shows the physicochemical properties of each branched CD obtained.

1 of each substance? Table 2 shows the results of investigating how many d of water dissolves in water.

α-CD 5.7β-CD
53.3r-CD
3.8 As is clear from Table 2, glucosylated branched CD was found to have extremely high solubility in water compared to α, β, and r-CD.

Blood was collected from a healthy adult female volunteer using a syringe containing cyheparin, and approximately 10 times the volume of physiological saline was added to the blood and suspended for 3,000 ml.
Centrifugation was carried out at rpm for 5 minutes, the upper groove was discarded, and the same operation was repeated twice, and an appropriate amount of physiological saline was added to the resulting blush spheres to obtain a 2-chi blush sphere suspension. 2% red blush suspension 0. IWLl was added to various CD solutions 31rLl and incubated at 37°C for 1 hour.

After the reaction, centrifugation was performed, and the absorbance of the supernatant at 540 nm was measured. The hemolysis rate was expressed as a relative hemolysis rate, with the hemolysis rate in physiological saline being 0 cm and the hemolysis rate in distilled water for injection being 100 cm. The results are shown in Figure 1.

As shown in Fig. 1, branched CD is different from methylated derivatives, and its hemolytic property was found to be equal to or lower than that of CD, indicating that this substance is highly safe.

A fixed excess amount of poorly water-soluble drug was added to 11 L of each 1.5 x 10 M CD solution, shaken at 30°C until equilibrium was reached, passed through ν, and the content of each drug dissolved in solution F was analyzed by high-performance liquid chromatography. (HPLC) Measurement for each CD
The solubilization ability of Table 3 shows the results.

As is clear from Table 3, glucosylated branched CD, % branched β-CD (Gβ-CD, 2Gβ-CD) has a high ability to solubilize various poorly soluble drugs, and in particular produces precipitated clathrates. It became clear that there were no characteristics.

EXAMPLES Next, the present invention will be explained in more detail with reference to Examples, but the present invention is not limited to these Examples in any way.

Example 1 Cosylated branched CD, G prepared in Experimental Example (1)
When 10ml of physiological saline was added to β-CD901!9 and estriol 20~ (1:1 molar ratio), and the mixture was thoroughly mixed at room temperature and the inclusion operation was performed, a completely transparent liquid was obtained. . After freeze-drying the heavy liquid, an appropriate amount of lactose was added and a tablet was prepared using a tablet machine. Distilled water 1. After adding Q ml and stirring thoroughly, add the estriol dissolved in water to U.
When quantified by V absorption measurement, it was 2606 μf/lut. Separately, when the solubility of estriol in water was measured in a similar preparation using estriol and lactose, it was found to be 29μ? -/ml, a remarkable solubilizing effect of Gβ-CD was observed.

Example 2 Glucosylated branched CD, 2G prepared in Experimental Example (1)
10 times of physiological saline was added to β-CD901!9 and estriol 201q (molar ratio 1:1), and the solubility of estriol in water was examined in the same manner as in Experimental Example 1. The first 10 d of liquid containing 2Gβ-CD and estriol was completely dissolved, and in the solubility test from the tablet, 2517 μl”/It O111 degrees of estriol was solubilized in water.
2Gβ-CD was also shown to have a solubilization ability comparable to that of Gβ-CD.

Example 3 Experimental example (1) Branched CD prepared by K, Gβ-CD0.1
7 capes and 0.10 q of cytoxin (molar ratio 1:1) to 1.0 q of physiological saline. When Od was added and the mixture was brought to room temperature and thoroughly stirred, the liquid became completely transparent and solubilized. After freeze-drying the liquid obtained in the same manner as in Example 1, an appropriate amount of lactose was added and tableted.
The solubility of cytotoxin in this tablet in water was examined using a small amount of water and found to be 5834μ, P/G (at 25°C).

Example 4 Branched CD prepared in Experimental Example (1), 2Gβ-CDO6
Liquid preparations and tablets similar to those in Example 3 were prepared using Example 19 and digitoxin 0.1. The obtained liquid was completely transparent. The water solubility of digitoxin from the tablets was 5252 μIP/rlLl.

When liquid preparations and tablets were prepared using only digitoxin in the same manner as in Examples 3 and 4, digitoxin was hardly dissolved in the liquid preparation, and the solubility of digitoxin from the tablets was only 5.0 μf/ml.

Example 5 Glucosylated branched CD prepared in Experimental Example (1).

Gβ-CD560W and phenoparpital 100w
Distilled water IQd was added to 9 (molar ratio 1:1), and 9 was heated at room temperature.
The mixture was sufficiently stirred and kneaded for 0 minutes to achieve inclusion.

After drying this, an appropriate amount of lactose was added, and the mixture was made into tablets using a tablet machine. The solubility of 7-enoparbital from this tablet in water was determined in the same manner as in Example 1, and was found to be 4466 μm at 25°C. / It was. The solubility of phenoparbital alone in water is 1350 μf/vtl, and Gβ-
The effect of solubilization by CD was confirmed.

Example 6 Glucosylated branched CD, Gβ prepared in Experimental Example (1)
- CDI 7”9 and vitamin D, 5 capes (molar ratio 1
: 10 tJ of physiological saline was added to 1) and stirred sufficiently at room temperature to carry out an inclusion operation, resulting in a completely transparent solution. After freeze-drying the heavy liquid in the same manner as in Example 1, an appropriate amount of lactose was added to prepare tablets. When the solubility of vitamin D3 in this tablet in water was measured, it was found that s 1s AP/+ at 25°C.
It was a/. Vitamin D.

alone did not dissolve in water at all.

Example 7 Branched CD prepared in Experimental Example (1), 2Gβ-CD19
A liquid preparation and a tablet were prepared in the same manner as in Example 6 using Misaki and 35 gg of vitamin D (molar ratio, 1:1).

The obtained liquid was completely transparent. The solubility of vitamin D3 from the tablet in water was 523 μp/m/O. Since vitamin D3 does not dissolve in water at all, Gβ-C
The effect on solubilization of D and 2Gβ-CD was confirmed.

[Brief explanation of the drawing]

FIG. 1 shows the results of a hemolytic test on various cyclodextrins.

Claims (1)

[Scope of Claims] 1. A composition in which a water-insoluble or poorly water-soluble pharmacologically active substance and a glucosylated branched cyclodextrin coexist. 2. The composition according to claim 1, wherein the water-insoluble or poorly water-soluble pharmacologically active substance is a steroidal anti-inflammatory agent, a cardiotonic agent, a central nervous system depressant, an antibiotic, or a fat-soluble vitamin. 3. Glucosylated branched cyclodextrin is α-, β-
Or, 1 to 3 glucoses are attached to γ-cyclodextrin via the oxygen atom at the 6-position of the cyclodextrin.
, 1→6 bond.