JP2000191704A - Polyphosphorylated cyclodextrin, its production and use - Google Patents

Abstract

PROBLEM TO BE SOLVED: To produce a physiologically acceptable new polyphosphorylated cyclodextrin having improved solubility in water and useful as a solubilizing agent or the like of a medicine or the like. SOLUTION: This new polyphosphorylated cyclodextrin is a compound of formula I Rx is a phosphate group of the formula [PO(OR4)]j-O-PO(OR4)(OR5) ((j) is 1-6; R4 and R5 are each H, an alkali metal ion or the like), H, methyl or the like; with the proviso that at least one of Rx is the phosphate group; R1 is H, methyl, (2-hydroxy)propyl, glucosyl or the like; R2 and R3 are each H, methyl, (2-hydroxy)propyl or phosphate group; l+m is 6-8), e.g. a polyphosphorylated α-cyclodextrin. Preferably, the compound of formula I is obtained, for example, by reacting a cyclodextrin of formula II with a phosphorylating reagent such as phosphorus pentoxide and phosphorus oxychloride in the presence of an amide-based solvent or an ether-based solvent.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a polyphosphorylated cyclodextrin, its production method and use, and more particularly to a novel polyphosphorylated cyclodextrin, its production method, and a solubilizer containing the compound as an active ingredient.

[0002]

2. Description of the Related Art Cyclodextrin or a complex thereof having no substituent introduced therein has some disadvantageous properties such as low water solubility and low solubilizing power. In order to improve these disadvantageous properties, it is necessary to substitute one or more hydroxyl groups of the constituent anhydrous glucopyranose units. Some cyclodextrins having a substituent introduced therein are stable under physiological conditions, and some are unstable under physiological conditions. The chemically stable substituent is usually an ether type substituent, and the chemically unstable substituent is usually an organic or inorganic ester type substituent.

[0003] Lower alkyl ether derivatives, such as alkylated or (hydroxy) alkylated derivatives, have very high water solubility, high solubilizing power and characteristics as a water-soluble complex. However, relatively high chemical stability can cause severe physiological difficulties. Specific examples include hemolytic activity (methylcyclodextrin), harmful effects on the kidney ((hydroxy) alkylated derivatives), and the like. Chemically labile compounds, on the other hand, when administered over a long period of time, may cause the appearance of currently unknown effects due to the accumulation of the substance. For example, the formation of a neutral ester, such as an acetate, cannot solve the problem of the original molecule because the original cyclodextrin is regenerated by hydrolysis in vivo.

In order to develop cyclodextrin derivatives having good water solubility, high solubilizing power and low accumulation when administered over a long period of time,
It is considered that the cyclodextrin is sulfoalkylated or sulfated to give a charged cyclodextrin derivative. However, methods for preparing sulfoalkylated cyclodextrins are difficult. Recently, a sulfated cyclodextrin, which is an anionic ester, has been proposed, but the field of application of this derivative is immunomodulation, and its water solubility is good, but its solubilizing power can be satisfied in many cases. Not something.

[0005]

The main object of the present invention is to develop a novel substance which is physiologically acceptable and can be used as a solubilizer for pharmaceuticals and the like. Further, it is an object of the present invention to establish a method for producing the substance.

[0006]

Means for Solving the Problems The present invention according to claim 1 is a polyphosphorylated cyclodextrin represented by the following general formula (I).

Embedded image

(Where R^X At least one of-[PO
(OR^Four)]_J -O-PO (OR^Four) (OR^Five)
Acid groups and other R^X Is phosphate group, hydrogen atom, methyl
Group or a (2-hydroxy) propyl group
R¹ Represents a hydrogen atom, a methyl group, (2-hydroxy)
Ropyl, glucosyl, galactosyl, maltosyl
Glucosyl group or galactosyl group into which
Or a maltosyl group;^Two And R
^Three Represents a hydrogen atom, a methyl group, (2-hydr
Loxy) a propyl group or a phosphate group,
+ M is any of 6 to 8, and J is an integer of 1 to 6.
You. In addition, the other R^X And R¹ , R^Two , R ^Three In
The phosphate group has an average number of phosphorus atoms of 2 to 12 in one molecule.
, -PO (OR^Four) (OR^Five) Or-[PO (O
R^Four)]_J -O-PO (OR^Four) (OR^Five) Where
J is an integer of 1 to 6, R^Four And R^Five Is water separately or simultaneously
Elemental atom, alkali metal ion, alkaline earth metal ion
Or a substituted or unsubstituted ammonium ion
You. )

The present invention according to claim 2 is that the cyclodextrin is α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, (2-hydroxy) propyl-cyclodextrin, methyl-cyclodextrin, glucosyl-cyclodextrin. The polyphosphorylated cyclodextrin represented by the general formula (I) according to claim 1, which is any one of maltosyl-cyclodextrin and galactosyl-cyclodextrin.

According to a third aspect of the present invention, a phosphate esterification reagent is reacted with a cyclodextrin represented by the following general formula (II). This is a method for producing a polyphosphorylated cyclodextrin represented by the formula:

Embedded image

(Wherein R¹ Represents a hydrogen atom, a methyl group, (2
-Hydroxy) propyl group, glucosyl group, galactosyl
Group, maltosyl group, -PO (OR^Four) (OR^Five) And-
[PO (OR^Four)]_J -O-PO (OR^Four) (OR^FiveNo)
Or R^Two And R^Three Is a hydrogen atom simultaneously or separately,
Methyl group, (2-hydroxy) propyl group, -PO (O
R ^Four) (OR^Five) And-[PO (OR^Four)]_J -O-PO (O
R^Four) (OR^Five). J is an integer of 1 to 6.
A number, R^Four And R^Five Is a hydrogen atom separately or simultaneously,
Alkali metal ions, alkaline earth metal ions or
Indicates a substituted or unsubstituted ammonium ion. )
You.

According to a fourth aspect of the present invention, the phosphate esterifying reagent is selected from the group consisting of phosphorus pentoxide, phosphorus oxychloride, alkali metal phosphate, hydrogen phosphate, alkyl chloride phosphoride and active phosphoric acid. A method according to claim 3.

The present invention according to claim 5 is the process according to claim 3, wherein the reaction is carried out in the presence of an amide solvent or an ether solvent.

According to a sixth aspect of the present invention, there is provided a solubilizing agent containing the polyphosphorylated cyclodextrin represented by the general formula (I) according to the first aspect.

[0014]

BEST MODE FOR CARRYING OUT THE INVENTION The above-mentioned polyphosphorylated cyclodextrin represented by the general formula (I) is produced by reacting a cyclodextrin represented by the general formula (II) with a phosphorylation reagent. Can be. The cyclodextrin used in the present invention generally has α
-, Β-, γ-cyclodextrin, (2-hydroxy) propyl- or methyl-cyclodextrin, but a cyclodextrin having a branched structure, for example, glucosyl-, maltosyl-, galactosyl-cyclodextrin, or a branched cyclodextrin. A heterobranched cyclodextrin in which a mannosyl group is bonded to a hydroxyl group of a glucosyl group or a maltosyl group which is a side chain can also be used. Further, as shown in the general formula (II), those having a phosphate group introduced can be used.

Next, the phosphorylation reagent used for polyphosphorylating cyclodextrin includes phosphorus pentoxide, phosphorus oxychloride, alkali metal phosphate,
Examples thereof include hydrogen phosphate, alkyl chloride phosphoride, and activated phosphoric acid, and phosphorus pentoxide and phosphorus oxychloride are preferred.

The reaction for polyphosphorylating cyclodextrin can be carried out at room temperature (about 20 to 30 ° C.), but it is preferable to use a reaction mixture cooled in advance. This reaction is preferably performed in the presence of a solvent. Examples of the solvent include amide solvents such as formamide, N, N-dimethylformamide, N, N-dimethylacetamide, and hexamethylene phosphorylamide, and 1,4-dioxane. Ether solvents such as Of these, amide solvents such as N, N-dimethylformamide are preferred.

After completion of the reaction, the desired reaction product is isolated and purified by a conventional method. For example, after washing with an organic solvent such as n-hexane, the product is dissolved in water, and the solution is cleaned with activated carbon. Then, it freeze-drys and obtains a target compound. The product may be in a free form, but the product can be converted to a salt form if desired. For example, various salts are formed by dissolving a crude product obtained by filtration from the reaction mixture in water and adjusting the pH of the aqueous solution to a predetermined value. Salts that can be used in the present invention include:
Alkali metal salts such as sodium salt, potassium salt and lithium salt, alkaline earth metal salts such as calcium salt, magnesium salt and barium salt, and amines such as trimethylamine salt, triethylamine salt, ethanolamine salt and amino acid salt There are salts and the like.

The polyphosphorylated cyclodextrin of the present invention
Is represented by the aforementioned general formula (I). formula
Medium, R^X At least one of-[PO (OR^Four)]_J -O
-PO (OR^Four) (OR^Five) Is a phosphate group represented by
R^X Is a phosphate group, a hydrogen atom, a methyl group and
Roxy) propyl group;¹ Is hydrogen field
, Methyl group, (2-hydroxy) propyl group, gluco
A sil, galactosyl, maltosyl or phosphate group
Introduced glucosyl group, galactosyl group or mal
Any of the tosyl groups,^Two And R^Three Are at the same time or different
Hydrogen atom, methyl group, (2-hydroxy) propyl
L + m is from 6 to 8
J is an integer of 1 to 6. In addition, the other
R^X And R¹ , R^Two , R ^Three Phosphoric acid group is one molecule
The average number of phosphorus atoms in the compound is 2 to 12, and -PO (OR
^Four) (OR^Five) Or-[PO (OR^Four)]_J -O-PO
(OR^Four) (OR^Five) Where J is an integer from 1 to 6,
R^Four And R^Five Are separately or simultaneously hydrogen atoms, alkali gold
Genus ion, alkaline earth metal ion or substitution
Represents an unsubstituted ammonium ion. Where ammoni
The substituent of the um ion is C₀ ~ C₆ Alkane, C
_Two ~ C₆ Alkene or C_Two ~ C₆ Alkyne
These can be used separately or simultaneously with benzene and halogen
Benzene, nitro-substituted benzene, alkyl-substituted benzene
, Haloalkyl-substituted benzene, naphthalene, halogen
Substituted naphthalene, nitro substituted naphthalene, alkyl substituted
Substituted with naphthalene, haloalkyl-substituted naphthalene, etc.
May be used.

In the polyphosphorylated cyclodextrin of the present invention, the structural glucose unit (m) and the substituted glucose unit (l) into which the polyphosphate group has been introduced form a cyclic structure in an arbitrary order by bonding at the 1- and 4-positions. The following substances can be mentioned as typical examples. In addition, the thing containing 2-20 phosphorus atoms in a molecule is preferable.

[0020]

Embedded image

In the compound of the formula (III), at least one of R ^X represents a polyphosphate group. The average number of phosphorus atoms in one molecule, that is, DS, can be adjusted by the number of equivalents of the phosphorylating reagent to be added. DS is, elemental analysis or ³¹ P-
It is quantified by NMR and cannot give the exact number of substituted sugar hydroxyl groups, but it does well characterize the molecule. In the formula (III), when at least one of R ^X is a polyphosphate group (DS = 2 to 4) and R ^X and R ^Y other than the polyphosphate group are hydrogen atoms, this compound is 1+
When m is 6, polyphosphorylated α-cyclodextrin (DS = 2 to 4), 7 is polyphosphorylated β-cyclodextrin (DS = 2 to 4), and 8 is polyphosphorylated γ-cyclodextrin (DS). = 2-4).

In the formula (III), when at least one of R ^X is a polyphosphate group (DS = 6 to 12) and R ^X and R ^Y other than the polyphosphate group are hydrogen atoms, this compound is 1 + m
Are polyphosphorylated α-cyclodextrins (DS = 6 to 12), 7 are polyphosphorylated β-cyclodextrins (DS = 6 to 12), and 8 are polyphosphorylated γ-cyclodextrins (DS = 6-12)
It is.

In the formula (III), when at least one of R ^X is a polyphosphate group (DS = 2 to 4) and R ^X and R ^Y other than the polyphosphate group are a hydrogen atom or a (2-hydroxy) propyl group, The compound having l + m of 6 has polyphosphorylated (2-hydroxy) propyl α-cyclodextrin (DS = 2 to 4), and the compound of 7 has polyphosphorylated (2-hydroxy) propyl β-cyclodextrin (DS = 2 to 2). 4) and 8 are polyphosphorylated (2-hydroxy) propyl γ-cyclodextrins (DS = 2
To 4).

In the formula (III), at least one of R ^X is a polyphosphate group (DS = 6 to 12), and R ^X and R ^Y other than the polyphosphate group are a hydrogen atom or (2-hydroxy)
In the case of a propyl group, compounds having l + m of 6 are polyphosphorylated (2-hydroxy) propyl α-cyclodextrin (DS = 6 to 12), and those of 7 are polyphosphorylated (2-hydroxy) propyl β-cyclodextrin. (DS = 6 to 12) and 8 were polyphosphorylated (2-
Hydroxy) propyl γ-cyclodextrin (DS
= 6 to 12).

In the formula (III), when at least one of R ^X is a polyphosphate group (DS = 2 to 4) and R ^X and R ^Y other than the polyphosphate group are a hydrogen atom or a methyl group, this compound has l + m 6 are polyphosphorylated methyl α-cyclodextrins (DS = 2 to 4), and 7 are polyphosphorylated methyl β-cyclodextrins (DS = 2 to 4).
4) and 8 are polyphosphorylated methyl γ-cyclodextrins (DS = 2 to 4).

In the formula (III), when at least one of R ^X is a polyphosphate group (DS = 6 to 12) and R ^X and R ^Y other than the polyphosphate group are a hydrogen atom or a methyl group, this compound has l + m 6 are polyphosphorylated methyl α-
Cyclodextrins (DS = 6 to 12) and 7 were polyphosphorylated methyl β-cyclodextrins (DS =
6 to 12) and 8 are polyphosphorylated methyl γ-cyclodextrins (DS = 6 to 12).

When the polyphosphorylated cyclodextrin or a salt thereof of the present invention is used as a pharmaceutical preparation, it is administered orally or parenterally (eg, intravenously, intramuscularly, topically and subcutaneously). The form is arbitrary, and includes solutions, tablets, capsules, powders, injection products and the like. When the substance of the present invention is used as a carrier, the amount to be used differs depending on the solubilizing power and the dose of the drug to be administered. For example, when solubilizing testosterone, 10% polyphosphorylated β-cyclodextrin (D
Using S = 4), a solution having a concentration of 10.2 mg / cm ³ can be prepared and used.

[0028]

EXAMPLES The present invention will be described below in detail with reference to examples and the like, but the present invention is not limited thereto. Example 1 Preparation of polyphosphorylated α-, β- or γ-cyclodextrin (DS = 2-4) 0.01 M dry α-, β- or γ-cyclodextrin (9.7 g, 11.4 g, 13 respectively) .0 g) to 35
cm ³ of N, N-dimethylformamide (hereinafter DMF)
Abbreviated, stored on CaH ₂ powder)
The solution or suspension was cooled below 20 ° C. Then
0.07M (9.5 g) of powdered phosphorus pentoxide was added to DMF1
The suspension in 10 cm ³ was added and mixed. The reaction mixture becomes almost clear within minutes and the temperature is 60 ° C.
The mixture was stirred for 1 day without cooling. Several hours after the start of stirring, it became somewhat difficult to continue stirring, but stirring was continued overnight.

The precipitate formed is filtered, and the reaction vessel is cooled to 20
The precipitate was washed twice with ³ cm ³ of dry DMF and 4 times with 50 cm ³ of n-hexane, and the precipitate adhering to the container was recovered.
In this way, a very hygroscopic crystalline substance was obtained, which was added to 500 cm ³ of deionized water. The crystalline material added to the deionized water was initially cloudy, but became a homogeneous solution within 3 hours. This solution is clarified with 10% by weight of activated carbon,
The pH of the resulting colorless acidic solution was adjusted to a range of 6.50 to 6.75 using a saturated sodium hydrogen carbonate solution. Next, the solution was subjected to membrane filtration (porosity ≦ 40 μm) and then freeze-dried.

By the above operation, a solid was obtained. It is readily soluble in water but does not have a precisely determined melting point. The DS of the product is 3.5-4 per cyclodextrin ring and the ratio of mono-, di- and triphosphates is 1: 1 according to ³¹ P-NMR spectrum. : 1 to 3: 2: 1. The product is represented by the general formula (III), wherein l + m = 6 is polyphosphorylated α-cyclodextrin, 7 is polyphosphorylated β-cyclodextrin, and 8 is polyphosphorylated γ-cyclodextrin. is there. Table 1 shows the analysis results of these products. The free phosphate was 10% by weight, and the yield was calculated on the assumption that the conversion of cyclodextrin monophosphate to the sodium salt was completed. In the table, α-CD is polyphosphorylated α-cyclodextrin, β-CD is polyphosphorylated β-cyclodextrin, and γ-CD is polyphosphorylated γ.
-Cyclodextrins are indicated respectively.

[0031]

[Table 1] Table 1

Example 2 Polyphosphorylated α-, β- or γ
Preparation of cyclodextrin (DS = 6-12) 35 of 0.01M dry α-, β- or γ-cyclodextrin (9.7 g, 11.4 g, 13.0 g respectively).
It was dissolved or suspended in cm ³ of DMF, and the solution or suspension was cooled with stirring to 20 ° C. or lower. To this was added 0.135M (1%) suspended in 95 cm ³ of DMF.
9.2 g) of phosphorus pentoxide powder was added. The reaction mixture became clear within minutes and the temperature rose to 60 ° C., but was stirred with a stirrer without cooling. Several hours after the start of stirring, it became difficult to continue stirring, but stirring was continued overnight without cooling.

The precipitate formed was filtered, and the reaction vessel was further washed twice with 35 cm ³ of dry DMF and 100 cm ³ of n-
After washing four times with hexane, the precipitate attached to the container was recovered. In this way, a very hygroscopic crystalline substance was obtained, which was dissolved in 500 cm ³ of deionized water. The reaction solution immediately after adding the crystalline material to the deionized water became cloudy, but became a homogeneous solution within 3 hours. At this point, 1
Clarification was performed using 0% by weight of activated carbon, and the pH of the obtained colorless acid solution was adjusted to be in the range of 6.50 to 6.75 using a saturated sodium hydrogen carbonate solution. Next, the solution was subjected to membrane filtration (porosity ≦ 40 μm) and freeze-dried.

By the above operation, a solid was obtained. It is readily soluble in water but does not have a precisely determined melting point. The DS of the product is about 8-9 per cyclodextrin ring, and the ratio of mono-, di- and tri-phosphates is 1: 1: 1, according to the ³¹ P-NMR spectrum. It was between 1 and 2: 1: 1. The product is represented by the general formula (III), wherein l + m = 6 is polyphosphorylated α-cyclodextrin, 7 is polyphosphorylated β-cyclodextrin, and 8 is polyphosphorylated γ-cyclodextrin. is there. Table 2 shows the analysis results of these products. The symbols in the table are the same as those in Table 1.

[0035]

[Table 2] Table 2

Example 3 Preparation of polyphosphorylated (2-hydroxy) propyl α-, β- or γ-cyclodextrin (DS = 2-4) 0.01M dried (2-hydroxy) propyl α
-, Β- or γ-cyclodextrin (DS = 2.7
Hereinafter, 11.3 g, 13.0 g, and 14.5 g, respectively, where DS indicates the degree of introduction of (2-hydroxy) propyl group to all hydroxyl groups in cyclodextrin. Hereinafter the same) was dissolved or suspended in 25 cm ³ of DMF. After the solution or suspension was cooled to 20 ° C. or lower, 0.07M or less phosphorous pentoxide powder (9.5 g) suspended in 50 cm ³ of DMF was added.

The reaction mixture becomes clear within minutes,
The temperature rose to 60 ° C., but was stirred for one day without cooling. After stirring was continued overnight, 30 cm ³ of anhydrous ethyl ether was added to form a precipitate. The precipitate was filtered, and the reaction vessel was further washed twice with 20 cm ³ of anhydrous ethyl ether and four times with 50 cm ³ of n-hexane to collect the precipitate adhering to the vessel. In this way, a very hygroscopic crystalline material was obtained, which was added to 250 cm ³ of deionized water. Immediately after the crystalline material was added to deionized water, it became cloudy, but became a homogeneous solution within 3 hours. When the reaction solution became homogeneous, it was clarified with 10% by weight of activated carbon, and the pH of the obtained colorless acidic solution was adjusted.
Using saturated sodium bicarbonate solution for 6.50-
It adjusted so that it might be in the range of 6.75.

Subsequently, the pH-adjusted solution was added to about 150 c
Concentrate until it reaches m ³ , then add ethanol 600 cm ³
To give a waxy precipitate. This precipitate was dissolved in 100 cm ³ of deionized water, and when a waxy precipitate formed again, 500 cm ³ of ethanol was added. This solution is redissolved in deionized water, and the solution is subjected to membrane filtration (porosity ≦ 4).
0 μm) and freeze-dried.

The DS of the product is about 3 to 4 per cyclodextrin ring, and the ratio of mono-, di- and triphosphates according to the spectrum of ³¹ P-NMR is: It was about 2: 1: 1. In addition, free phosphorus oxide was 15% by weight, and the yield was calculated on the assumption that monophosphate of cyclodextrin was completely converted to a sodium salt. Table 3 shows the measurement results. The symbols in the table are the same as those in the first table of the first embodiment. The product is represented by the above general formula (III), wherein l + m = 6 is polyphosphorylated (2-hydroxy) propyl α-cyclodextrin, and 7 is polyphosphorylated (2-hydroxy) propyl β-cyclodextrin. , 8 are polyphosphorylated (2-hydroxy) propyl γ-cyclodextrins.

[0040]

[Table 3] Table 3

Example 4 Preparation of polyphosphorylated (2-hydroxy) propyl α-, β- or γ-cyclodextrin (DS = 6-12) 0.01 M dried (2-hydroxy) propyl α
-, Β- or γ-cyclodextrin (DS = 2.7
Hereinafter, 11.3 g, 13.0 g, and 14.5 g) were added to 2
It was dissolved or suspended in 5 cm ³ of DMF, and the solution or suspension was cooled to below 20 ° C. with stirring. To this was added 0.135 m of phosphorus pentoxide powder (19.2 g) suspended in 110 cm ³ of DMF.

The reaction mixture is almost transparent within a few minutes.
Became clear and the temperature rose to 60 ° C, but cooling
Without stirring for 1 day. After that, stirring was continued overnight.
30cm later^Three Of diethyl ether to precipitate
Generated. The formed precipitate is filtered and the reaction volume is
20cm^Three 50 cm twice with diethyl ether ^Three 
Washed four times with n-hexane, and the precipitate adhering to the container
The thing was collected. The resulting highly hygroscopic crystalline material
Deionized water 200cm^Three Was dissolved. Subsequent operations
Was carried out in the same manner as in Example 3, and
Oxide was obtained.

The product polyphosphorus oxide has a DS of 6 to
12, and the ratio of mono-, di- and triphosphates is about 2: according to ³¹ P-NMR spectrum.
1: 1. In addition, free phosphorus oxide is 15% by weight.
Below, yields were calculated assuming that the monophosphate of cyclodextrin was completely converted to the sodium salt. Table 4 shows the measurement results. The symbols in the table are the same as those in the first table of the first embodiment. The product is represented by the above general formula (III), wherein l + m = 6 is polyphosphorylated (2-hydroxy) propyl α-cyclodextrin, and 7 is polyphosphorylated (2-hydroxy) propyl β-cyclodextrin. , 8 are polyphosphorylated (2-hydroxy) propyl γ-cyclodextrins.

[0044]

[Table 4] Table 4

Example 5 Polyphosphorylated methyl α-, β-
Or preparation of γ-cyclodextrin (DS = 2-4) 0.01M dried methyl α-, β- or γ-cyclodextrin (DS = 13 or less, 11.5 g each,
3.2 g, 15.0 g) are dissolved or suspended in 25 cm ³ of DMF and the solution or suspension is stirred at 20 ° C.
It cooled until it became below. Add 50cm ³ DMF
0.07M or less phosphorus pentoxide powder (9.5
g) was added.

The reaction mixture became almost clear within minutes and the temperature rose to 60 ° C., but was stirred for one day without cooling. When stirring was continued overnight, a viscous precipitate formed. 20 cm ³ of n-hexane was added to the formed precipitate, and the formed solid was filtered. Further, the reaction vessel was washed twice with 20 cm ³ of anhydrous ether for 50 times.
The precipitate was washed four times with cm ³ of n-hexane, and the precipitate attached to the container was recovered. The solid thus obtained was dissolved in 200 cm ³ of deionized water. All subsequent operations were performed in the same manner as in Example 3 to obtain the target polyphosphorus oxide.

The DS of the above product is about 2 to 4 and the ratio of mono-, di- and triphosphates is ³¹ P
According to -NMR spectrum, it was about 2: 1: 1. In addition, free phosphorus oxide is 10% by weight or less,
Yields were calculated assuming that the monophosphate of cyclodextrin was completely converted to the sodium salt. Table 5 shows the measurement results. The symbols and the like in the table are the first in the first embodiment.
Same as the table. The product is represented by the above general formula (III), wherein l + m = 6 is polyphosphorylated methyl α-cyclodextrin, 7 is polyphosphorylated methyl β-cyclodextrin, and 8 is polyphosphorylated methyl γ
-Cyclodextrin.

[0048]

[Table 5] Table 5

Example 6 Polyphosphorylated methyl α-, β-
Or preparation of γ-cyclodextrin (DS = 6 to 12) 0.01M dried methyl α-, β- or γ-cyclodextrin (DS = 13 or less, 11.5 g each,
3.2 g, 15.0 g) was dissolved or suspended in 25 cm ³ of DMF, and the solution or suspension was cooled to 20 ° C. or lower. Add 110 cm ³ of DM to the solution or suspension.
F suspended in 0.135 M phosphorous pentoxide powder (19.2
g) was added.

The reaction mixture became almost clear within minutes and the temperature rose to 60 ° C., but was stirred for one day without cooling. When stirring was continued overnight, a viscous precipitate formed. After adding 20 cm ³ of n-hexane to the resulting precipitate, the resulting solid was filtered. Further, the reaction vessel was washed twice with 20 cm ³ of anhydrous ether for 50 times.
The precipitate was washed four times with cm ³ of n-hexane, and the precipitate attached to the container was recovered. The obtained solid is deionized water 10
0 cm ³ , and all subsequent operations were performed in Example 3.
In the same manner as described above, a target polyphosphorus oxide was obtained.

The DS of the product is about 6-9 and the ratio of mono-, di- and triphosphates is ³¹ P-NM
According to the spectrum of R, it was about 2: 1: 1. The free phosphate was less than 10% by weight, and the yield was calculated assuming that the monophosphate of cyclodextrin was completely converted to the sodium salt. Table 6 shows the measurement results. The symbols in the table are all the same as those in the first table of the first embodiment. The product is represented by the above general formula (III), wherein l + m = 6 is polyphosphorylated methyl α-cyclodextrin, 7 is polyphosphorylated methyl β-cyclodextrin, and 8 is polyphosphorylated methyl γ
-Cyclodextrin.

[0052]

[Table 6] Table 6

Test Example 1 Solubility test of ketoconazole Polyphosphorylated α- with DS = 4 prepared in Example 1
A 10% aqueous solution of cyclodextrin was prepared, and an excess amount of ketoconazole was added thereto. The suspension was stirred at 25 ± 1 ° C. for 24 hours, and then filtered using a cellulose nitrate membrane filter (pore diameter: 0.45 μm). The absorbance at 280 nm of the obtained filtrate was measured by ultraviolet photometry, and the solubility was calculated. As a control, a similar test was conducted using a 3% aqueous solution of α-cyclodextrin.

As a result, the solubility in water was 0.02 mg /
When the ketoconazole having a cm ³ was included in polyphosphorylated α-cyclodextrin, the solubility in water was 1.1 mg / cm ³ , and the solubility was increased 55-fold. On the other hand, control α-cyclodextrin 3%
The solubility in water when included with an aqueous solution is 0.2 mg /
cm ³ and a sufficient increase in solubility could not be achieved.

Test Example 2 Solubility test of testosterone Polyphosphorylated β- with DS = 4 prepared in Example 1
A 10% aqueous solution of cyclodextrin was prepared and excess testosterone was added thereto at room temperature with vigorous stirring. After stirring for 2 hours, the suspension was treated in the same manner as in Test Example 1, and the solubility in water was measured. As a control, the same test was performed using a 1.5% aqueous solution of β-cyclodextrin.

As a result, the solubility in water was 0.03 mg /
The testosterone of cm ³ is included in polyphosphorylated β-cyclodextrin, so that its solubility in water becomes 10.2 mg / cm ^{3 at} pH 6.0.
Solubility increased 340-fold. On the other hand, when included in a 1.5% aqueous solution of β-cyclodextrin, the solubility in water is 0.15 mg / cm ³ , and the solubility is only 5 mg / cm ^3.
There was only a doubling, with little improvement.

Test Example 3 Solubility test of furosemide Polyphosphorylated γ- of DS = 4 prepared in Example 1
A 10% aqueous solution of cyclodextrin was prepared, and an excessive amount of furosemide was added thereto while stirring vigorously at room temperature in the same manner as in Test Example 2, and the suspension was treated in the same manner to reduce the solubility of furosemide. It was measured. As a control, a similar test was performed using a 10% aqueous solution of γ-cyclodextrin.

As a result, the solubility in water was 0.06 mg /
By embedding furosemide, which is cm ³ , with polyphosphorylated γ-cyclodextrin, the solubility in water is:
At pH 6.2, the concentration was 2.5 mg / cm ³ , and the solubility was increased about 40-fold. On the other hand, when included in a 10% aqueous solution of γ-cyclodextrin, the solubility in water was 0.2 mg / cm ³ , and the solubility in water increased only about three times.

Test Example 4 Solubility test of ibuprofen DS = 4 polyphosphorylated β-prepared in Example 1
A 10% aqueous solution of cyclodextrin was prepared, and an excess amount of ibuprofen was added thereto in the same manner as in Test Example 2 to obtain a suspension. Then, the solubility of ibuprofen was measured by the same method. As a control, the same test was performed using a 1.5% aqueous solution of β-cyclodextrin.

As a result, the solubility in water was 0.08 mg /
Inclusion of cm ³ ibuprofen with polyphosphorylated β-cyclodextrin resulted in a water solubility of 9.8 mg / cm ^{3 and a} 123-fold increase in solubility. On the other hand, the solubility in water when included in a 1.5% aqueous solution of β-cyclodextrin is 1.7 mg / c.
It only showed m ³ .

Test Example 5 Solubility test of terfenadine Polyphosphorylated β-DS = 4 prepared in Example 1
A 10% aqueous solution of cyclodextrin was prepared, and an excess amount of terfenadine (Terfenadine) was prepared in the same manner as in Test Example 2.
adine) was added to obtain a suspension, and the solubility of terfenadine was measured in the same manner. In addition, as a control,
The same test was conducted using a 1.5% aqueous solution of β-cyclodextrin.

As a result, the solubility in water was 0.01 mg /
Inclusion of terfenadine in cm ³ with polyphosphorylated β-cyclodextrin resulted in a water solubility of 6.0 mg / cm ^{3 at} pH 6.1, increasing the solubility by a factor of 600. On the other hand, the solubility in water when included in a 1.5% aqueous solution of β-cyclodextrin was only 0.08 mg / cm ^3, and the effect of improving the solubility was not sufficient.

Test Example 6 Solubility test of astemizole Polyphosphorylated β- with DS = 4 prepared in Example 1
A 10% aqueous solution of cyclodextrin was prepared, and an excess amount of astemizole (Astemisol) was prepared in the same manner as in Test Example 2.
After the addition of zole), the solubility of astemizole was measured in the same manner as in Test Example 2. In addition,
As a control, the same test was performed using a 15% aqueous solution of β-cyclodextrin.

As a result, the solubility in water was 0.03 mg /
Inclusion of astemizole of cm ³ with polyphosphorylated β-cyclodextrin resulted in a solubility in water of 2.5 mg / cm ^{3 at} pH 6.3 and an increase of about 80 times in water. . On the other hand, when included in a 15% aqueous solution of β-cyclodextrin, the solubility in water was 0.08 mg / cm ³ , indicating that the solubility was hardly improved.

Test Example 7 Solubility test of dixoline A 10% aqueous solution of polyphosphorylated α-cyclodextrin having a DS of 8 to 9 prepared in Example 2 was prepared.
After adding an excess amount of dixolin to this, treatment was carried out in the same manner as in Test Example 1, and the solubility of dixolin was measured. As a control, one of α-cyclodextrin was used.
The same test was conducted using a 0% aqueous solution.

As a result, the solubility in water was 0.07 mg /
The inclusion in water of polyoxophosphoric acid α-cyclodextrin by the inclusion of dixoline of cm ³ ,
When the pH is 7.0 to 7.5, it becomes 3.6 mg / cm ³ ,
Solubility in water increased about 50-fold. On the other hand, the solubility in water when included in a 10% aqueous solution of α-cyclodextrin is 0.34 mg / cm ^{3 at} pH 7.2.
Thus, the solubility in water was not sufficiently improved.

Test Example 8 Solubility test of β-indoleacetic acid A 10% aqueous solution of polyphosphorylated β-cyclodextrin having a DS of 8 to 9 prepared in Example 2 was prepared, and an excess amount of β-indoleacetic acid was prepared. β-indoleacetic ac
id), and β
The solubility of indoleacetic acid was determined. As a control, the same test was performed using a 10% aqueous solution of β-cyclodextrin.

As a result, the solubility in water was 1.8 mg / c
By inclusion of β-indole acetic acid of m ³ with polyphosphorylated β-cyclodextrin, the solubility in water is 14.0 mg / cm ³ at pH 3.0,
The solubility in water was increased by about 8 times. On the other hand, when included in a 10% aqueous solution of β-cyclodextrin, the solubility in water was 3.6 mg / cm ³ , indicating that the solubility in water was not significantly improved.

Test Example 9 Solubility test of β-indoleacetic acid A 10% aqueous solution of polyphosphorylated γ-cyclodextrin having a DS of 8 to 9 prepared in Example 2 was prepared, and an excess amount of β-indoleacetic acid was added thereto. In addition, Test Example 1
The solubility of β-indoleacetic acid was measured according to the method described in (1). As a control, a similar test was performed using a 10% aqueous solution of γ-cyclodextrin.

As a result, the solubility in water was 1.8 mg / c
By inclusion of m ³ β-indoleacetic acid with polyphosphorylated γ-cyclodextrin, the solubility in water is 13.2 mg / cm ³ at pH 3.0,
Solubility in water increased about 7-fold. On the other hand, γ-
The solubility when included in a 10% aqueous solution of cyclodextrin was 3.3 mg / cm ³ , and the solubility was only doubled, and a sufficient improvement effect could not be achieved.

[0071]

According to the present invention, a polyphosphorylated cyclodextrin is provided. Since this substance has improved solubility in water and is physiologically acceptable, it can be used as a solubilizing agent for pharmaceuticals and the like. Furthermore, the present invention also provides a method for producing the substance.

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Katsuyoshi Nakanishi 13-46, Ogurocho, Tsurumi-ku, Yokohama, Kanagawa Prefecture Inside the Yokohama International Biotechnology Research Institute, Inc. (72) Inventor Jin Hashimoto 13-46, Ogurocho, Tsurumi-ku, Yokohama, Kanagawa Prefecture Inside Yokohama Institute of Biotechnology Co., Ltd. (72) Inventor Lathro Idinsky Hungary, 1025 Budapest, Pusta Seri Ut 59 Cyclolap Limited (72) Inventor Eva Fembesi Hungary, 1025 Budapest, Pusta Seri・ Uto 59 Cyclolap Limited (72) Inventor Joseph Cytori Hungary, 1025 Budapest, Pusta Seri Uto 59 Cyclolap Limited (72) Inventor Rios Cente Hungary, 1025 Budapest, Pusta - Uto, 59 Shikurorapu Limited in the F-term (reference) 4C076 AA12 CC31 EE39 FF15 FF67 4C090 AA02 AA05 AA09 BA10 BA11 BB04 BB12 BB32 BB36 BB52 BB64 BB85 BB96 BD02 BD36 BD42 CA38 DA09 DA23 4H050 AB68 BB15 BB20 BE04 BE54 BE90 WA13 WA15 WA19 WA23

Claims (6)

[Claims] 1. A polyline represented by the following general formula (I)
Oxidized cyclodextrin. Embedded image(Where R^X At least one of-[PO (OR^Four)]_J 
-O-PO (OR^Four) (OR^Five)
Other R^X Is a phosphate group, a hydrogen atom, a methyl group and (2
-Hydroxy) propyl group;¹ Is water
Element atom, methyl group, (2-hydroxy) propyl group,
Lucosyl group, galactosyl group, maltosyl group or phosphoric acid
Glucosyl group, galactosyl group or
A maltosyl group;^Two And R^Three Is also
Are independently hydrogen, methyl, (2-hydroxy) pro
A pill group or a phosphate group, and l + m is 6 to 8
And J is an integer of 1 to 6. Also before
Other R^X And R¹ , R^Two , R ^Three The phosphate group in is 1
The average number of phosphorus atoms in the molecule is 2 to 12,
(OR^Four) (OR^Five) Or-[PO (OR^Four)]_J -O-
PO (OR^Four) (OR^Five) Where J is an integer from 1 to 6.
Number, R^Four And R^Five Are separately or simultaneously hydrogen atoms,
Li metal ion, alkaline earth metal ion or substitution
Or an unsubstituted ammonium ion. ) 2. The method of claim 1, wherein the cyclodextrin is α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, (2-hydroxy) propyl-cyclodextrin, methyl-cyclodextrin, glucosyl-cyclodextrin, maltosyl-cyclodextrin, and galactosyl. The polyphosphorylated cyclodextrin represented by the general formula (I) according to claim 1, which is any one of -cyclodextrin. 3. A cyclone represented by the following general formula (II):
Reaction of dextrin with a phosphorylation reagent
And represented by the general formula (I) according to claim 1.
A method for producing polyphosphorylated cyclodextrin. Embedded image(Where R¹ Represents a hydrogen atom, a methyl group, (2-hydroxy
C) propyl group, glucosyl group, galactosyl group, mal
Tosyl group, -PO (OR^Four) (OR^Five) And-[PO (OR
^Four)]_J -O-PO (OR^Four) (OR^Five)
R^Two And R^Three Is simultaneously or separately a hydrogen atom, a methyl group,
(2-hydroxy) propyl group, -PO (OR ^Four) (OR
^Five) And-[PO (OR^Four)]_J -O-PO (OR^Four) (OR
^Five). J is an integer of 1 to 6,
R^Four And R^Five Are separately or simultaneously hydrogen atoms, alkali gold
Genus ion, alkaline earth metal ion or substitution
Represents an unsubstituted ammonium ion. ) 4. A phosphorylation reagent comprising phosphorus pentoxide,
4. The method according to claim 3, which is any one of phosphorus oxychloride, alkali metal phosphate, hydrogen phosphate, alkyl chloride phosphoride and activated phosphoric acid. 5. The method according to claim 3, wherein the reaction is carried out in the presence of an amide solvent or an ether solvent. 6. A solubilizer comprising the polyphosphorylated cyclodextrin represented by the general formula (I) according to claim 1.