JPS6310730A - Stable aqueous solution - Google Patents

Abstract

PURPOSE:An aqueous solution, containing a 4-carbamoyl-1-beta-D-ribofuranosyl- imidazolium-5-oleate, (hydrogen)sulfite, etc., having immunosuppressive action, diversified administration forms and improved stability. CONSTITUTION:An aqueous solution containing at least one selected from a sulfite, hydrogensulfite and metabisulfite in an aqueous solution containing a 4-carbamoyl-1-beta-D-ribofuranosyl-imidazolium-5-oleate or salt thereof (referred to as this compound). The total amount of the sulfite, hydrogensulfite and metabisulfite blended is 0.005-50pts.wt., preferably 0.05-2pts.wt. based on 1pts.wt. of this compound. The aqueous solution of this invention is preferably adjusted to 6-10pH and becomes usable as an eye drop or injection.

Description

Detailed Description of the Invention [Industrial Field of Application] The present invention relates to 4-carbamoyl-1-β-D-ribofuranosyl-imidazolium- which is a compound useful as a pharmaceutical.
Stable aqueous 5-oleate or its salt (hereinafter sometimes simply referred to as the present compound)? It is a night pill.

[Prior art]

4-Carbamoyl-1-β-D-ribofuranosyl-imidazolium-5-oleate, also called mizoribine, is produced by the filamentous fungus Eupenicillium brefeldiasum (Eupe
It is an imidazole-based nucleic acid-related compound discovered in the culture filtrate of Nicilium brefeldianu*), and is a substance that has immunosuppressive effects.

Its action is thought to competitively inhibit the pathway from inosinic acid to guanylic acid in the nucleic acid purine synthesis system, thereby selectively suppressing the proliferation of lymphocytes.

Tablets are already on the market for its powerful immunosuppressive effects.

By the way, as mentioned above, this compound has an immunosuppressive effect, so if it could be prepared into a highly concentrated and stable aqueous solution at around neutrality, it would be possible to administer it to the eye, and it would be useful for corneal transplantation. It can be used as a rejection inhibitor during surgery, and can also be used as an injection, making it possible to diversify its uses and administration forms.

[Problem that the invention seeks to solve]

From this point of view, the present inventors have conducted various studies on aqueous liquid preparations of the present compound, and have found that the present compound is unstable in an aqueous solution state.

Therefore, the object of the present invention is to provide an aqueous liquid preparation in which the present compound □ can be stably present, and as is clear from the above, the object of the present invention itself is a novel invention.

[Means for solving problems]

In order to solve this new object, the present inventors have attempted extensive studies on stabilizing the present compound in an aqueous solution.

As a result, the present inventors surprisingly found that by coexisting at least one selected from sulfite, hydrogen sulfite, and metabisulfite in an aqueous solution containing the present compound, an aqueous solution of the present compound can be obtained. However, even if a so-called stabilizer, which has traditionally been used to stabilize unstable substances in various aqueous liquids, is added, the stability of the drug will be significantly improved. The present invention was completed based on the discovery that a sufficient stabilizing effect could not be obtained for use as an anti-oxidant compound. That is, ascorbic acid, isoascorbic acid, which has been conventionally used as a stabilizer for various compounds;
Sodium carbonate, sodium citrate, disodium edetate, glycerin, polyvinylpyrrolidone, cyclodextrin (α-1β- and γ-), N-acetylcysteine, etc. do not contribute to the stabilization of this compound in aqueous solution. It has been found that the stability of the present compound or its salt in an aqueous solution is significantly improved when at least one selected from , sulfite, hydrogen sulfite and metabisulfite is contained.

That is, the present invention is an aqueous liquid preparation containing the present compound and at least one selected from sulfites, hydrogen sulfites, and metabisulfites.

The object to be stabilized in the present invention is mizoribine or a salt thereof, and examples of the salt include non-toxic inorganic acids or organic acids, or non-toxic organic bases or inorganic bases that can form a salt with the present compound.

In the aqueous solution of the present invention, at least one selected from sulfites, hydrogen sulfites and metabisulfites is contained in the aqueous solution containing the present compound. Examples of the salts in sulfite, hydrogen sulfite and metabisulfite include alkali metal salts such as sodium salts and potassium salts, alkaline earth metal salts such as calcium salts, and organic base salts such as ammonium salts. .

The aqueous liquid preparation of the present invention is not particularly limited as long as the present compound is dissolved in water, and examples include fIE forms such as eye drops and injections.

Examples of water include distilled water for injection, sterilized purified water, and the like.

The blending ratio of sulfite, hydrogen sulfite and metabisulfite to 1 part by weight of the present compound is usually o, oos to 50 parts by weight, preferably 0.05 to 2 parts by weight as a total amount thereof.

- The aqueous liquid preparation of the present invention can be produced by dissolving the present compound and at least one selected from sulfite, hydrogen sulfite, and metabisulfite in water, and the concentration of the present compound in the aqueous liquid preparation is against the water,
It is about 0.1 to 20 w/v%, preferably about 1 to 10 w/v%, and can be water-soluble to extremely high concentrations.

The amount of sulfite, hydrogen sulfite and metabisulfite added in the aqueous solution of the present invention is usually as follows:
It is about 0.1 to 5 w/v%, preferably about 0.5 to 2-/v%.

Furthermore, the present inventors obtained the following knowledge. That is, the present compound, as revealed in Experimental Example 1, contains all p)
I is unstable in aqueous solution of I, and relatively stable ρ
112 and around pH 7, it was found that this was still insufficient for practical use as a medicine, and as clarified in Experimental Example 3 below, the aqueous liquid preparation of the present invention was significantly effective at a pH of about 6 to 10. It was found to be stable. Therefore, the aqueous solution of the present invention has pi in -a.
(It is good to adjust the pH to about 6 to 10, preferably 117 to 9, and more preferably about 8 for three times.

The aqueous liquid preparation of the present invention includes, as long as it does not contradict the purpose of the present invention,
Additives normally used in aqueous solutions, such as buffers for pH adjustment (phosphate buffer, borate buffer, citrate buffer, tartrate buffer, acetate buffer, etc.), tonicity agents (sorbitol, glycerin, etc.) , polyethylene glycol, propylene glycol, glucose, sodium chloride, etc.), preservatives and disinfectants (benzalkonium chloride, paraoxybenzoic acid esters, benzyl alcohol, parachlormetaxinol, chlorcresol, phenethyl alcohol, sorbic acid or its salts, Thimerosal, chlorobutanol, etc.), chelating agents (sodium edetate, sodium citrate, condensed sodium phosphate, etc.), thickening agents (polyvinylpyrrolidone, methylcellulose, sodium carboxymethylcellulose, hydroxypropylcellulose, polyvinyl alcohol, sodium polyacrylate) etc.) can be added in the commonly used amounts.

The aqueous liquid preparation of the present invention may contain medicinal ingredients other than the present compound as long as they do not contradict the purpose of the present invention.

In preparing the aqueous solution of the present invention, the gt of the solution is
Depending on the type, per se known means can be adopted.

[Experiment example/Example]

Experimental Example 1 Add 20 g of water to 10 g of mizoribine and dissolve by heating.
60 ml of acetone are added and after standing at 5° C. for 24 hours it is filtered, washed with cold acetone and dried under reduced pressure for 24 hours. After repeating this operation twice, 100-methanol was added to prepare a suspension, which was refluxed for about 3 hours and filtered.
Wash with cold methanol. This was dried at 30°C under reduced pressure for 48 hours and used as a standard product (containing 199.5%: non-aqueous titration method).The decomposition rate constant for each pH of the standard product (
-1og k) is shown in FIG. 1.

As is clear from FIG. 1, this compound is most stable in aqueous solutions around pH 2 and 7.

Experimental Example 2 In order to clarify the stability improvement effect of mizoribine when at least one selected from sulfite, hydrogen sulfite, and metabisulfite is blended with mizoribine,
Stabilization tests were carried out using sodium sulfite, sodium hydrogen sulfite and sodium metabisulfite and as a control stabilizers commonly used to stabilize substances that are unstable in aqueous solutions. The stabilizers used as controls were ascorbic acid, isoascorbic acid (antioxidant), sodium carbonate (anticoloring agent), sodium citrate, disodium edetate, glycerin (decomposition inhibitor), and boric acid. , polyvinylpyrrolidone, cyclodextrin (α-1β- and γ-), and N-acetyl lastine.

The concentration of mizoribine was 3%, and the concentration of various stabilizers was 0.2.
．． 0.5.1.0%, and after storage at pH 7 and 40°C for 7 days, changes in color were observed. As a result, the groups to which sodium sulfite, sodium hydrogen sulfite, and sodium metabisulfite were added visually showed There was no appreciable coloration, whereas coloration was clearly observed in all of the control group.

Experimental Example 3 78 solutions were prepared by adding 3% mizoribine and 0.2% sodium sulfite to 0.1M phosphate buffer.

This solution was adjusted to various pH values listed in Table 1, and these were adjusted to 40%
After storage at ℃ for 7 days, the residual rate (%) of mizoribine was examined by high performance liquid chromatography.

As a control, a solution prepared by adding mizoribine at a concentration of 3% to 0.1 M phosphate buffer was used.

The results are shown in Table 1 below.

Experimental Example 4 The procedure of Experimental Example 3 was followed except that the concentration of sodium sulfite was 0.5%, and the residual rate of mizoribine was investigated. The results are shown in Table 2 below.

Experimental Example 5 The procedure of Experimental Example 3 was followed except that the concentration of sodium sulfite was 1.0%, and the residual rate of mizoribine was investigated. The results are shown in Table 3 below.

Experimental Example 6 The procedure of Experimental Examples 3 to 5 was followed except that sodium bisulfite was used instead of sodium sulfite in Experimental Examples 3 to 5, and the results shown in Tables 4 to 6 below were obtained.

Experimental Example 7 The same residual rate of mizoribine as in Experimental Examples 3 to 5 was obtained even if the treatment was performed according to Experimental Examples 3 to 5 except that metabisulfite was used instead of sodium sulfite in Experimental Examples 3 to 5. It will be done.

Formulation Example 1 (Eye drops) This compound 3.0g Sodium sulfite 1.0g Sodium hydrogen phosphate 0.5g Benzalkonium chloride
0.01g Sodium hydroxide Appropriate amount (pH 8,0) Sterile purified water All ffi
Sodium sulfite 1 in 100 m sterile purified water approximately 80 mF
．． 0g, disodium hydrogen phosphate 12 hydrate, 0.5g, and 3.0g of this compound were sequentially added and dissolved. To this solution, 0.01g of benzalkonium chloride was added and dissolved, and sodium hydroxide was further added to adjust the pH. The solution was adjusted to about 8 and sterilized purified water was added to make a total of 1i100-. This solution was sterilized by filtration (0.45 μrrt) and then aseptically filled into a plastic bottle for eye drops to obtain an eye drop.

Formulation Example 2 (injection) This compound 1.0g Potassium sulfite 0.5g Sodium hydrogen phosphate 0.5g Sodium chloride
0.3g sodium hydroxide appropriate amount (
pH 7,4) Sterile purified water Total amount: 100 - Sterile purified water about 80 - and potassium sulfite 0.5 g.

Add and dissolve 0.5 g of disodium hydrogen phosphate decahydrate and 1.0 g of this compound in sequence. Add sodium hydroxide to this solution to adjust the pH to 7.4, and add sterile purified water to make a total volume of 100 g. - This solution is sterilized by filtration (0,22
μm) and then aseptically filled into ampoules to obtain an injection.

Formulation example 3 (eye drops) This compound 3.0g Sodium metabisulfite 1.0g Sodium hydrogen phosphate 0.5g Benzalkonium chloride
0.01g sodium hydroxide appropriate amount (
Using a formulation with a pH of 8.0) sterile purified water (total amount: 100 - or higher), the eye drops were obtained by processing according to Formulation Example 1.

Formulation example 4 (eye drops) This compound 5.0g Sodium metabisulfite 1.0g Boric acid
0.5g Methyl paraoxybenzoate 0.026g Methyl paraoxybenzoate 0
．． 014g Sodium hydroxide appropriate amount (pH 8,
0) Sterile purified water total 1 10 (laj Sterilized purified water approximately 80111, methyl paraoxybenzoate 0
．． 026 g and propyl roseoxybenzoate 0.0
Add 14 g of sodium metabisulfite and dissolve while heating. To this solution, add 1. og.

Add 0.5 g of boric acid and 5.0 g of this compound one after another to dissolve, and then add sodium hydroxide to bring the ρ■ to about 8.0.
Sterilized purified water was added to bring the total volume to 100. This solution was sterilized by filtration (0.45 μff1) and then aseptically filled into a plastic bottle for eye drops to obtain an eye drop.

Formulation example 5 (eye drops) This compound 10.0 g Sodium thiosulfate 1.0 g Sodium hydrogen phosphate 0.5 g Thimerosal
0.0 O5g Sodium hydroxide Appropriate amount (pH 8,0) Sterile purified water Total amount 1
0〇-About 80% of sterile purified water and 1.0% of sodium thiosulfate
g, 0.5 g of disodium hydrogen phosphate and this compound 1
Add 0.0 g of thimerosal in sequence and dissolve. Add 5 g of thimerosal O, OO to the thus obtained solution and dissolve. Add sodium hydroxide to adjust the pH to approximately 8.0, and add sterile purified water to make a total of 1 tioo. I will dedicate it to you.

This liquid was filtered through 11t (0.45 μin) and then filled aseptically into a plastic bottle for eye drops to obtain an eye drop.

[Action/Effect]

The sulfite, hydrogen sulfite and metabisulfite used in the present invention have the effect of improving the stability of the present compound or its salt in an aqueous solution. The aqueous solution of the present invention containing at least one selected from bisulfites has the effect that the present compound and its salt can exist stably for a long period of time, is stable even at high concentrations, and can be used near neutrality. A stable aqueous solution can be obtained even when

(The following is a blank space) Table 1 Residual rate of mizoribine (%) In the table, Na means sodium salt (the same applies hereinafter).

Table 2 Mizoribine residual rate (%) Table 3: Mizoribine residual rate (%) Table 4: Mizoribine residual rate (%) Table 5: Mizoribine residual rate (%) Table 6: Mizoribine residual rate (%)

[Brief explanation of drawings]

FIG. 1 is a graph showing the pn profile of the stability of 4-carbamoyl-1-β-D-ribofuranosyl-imidazolium-5-oleate in an aqueous solution.

Claims (2)

[Claims] (1) An aqueous liquid preparation containing 4-carbamoyl-1-β-D-ribofuranosyl-imidazolium-5-oleate or a salt thereof, and at least one selected from sulfites, hydrogen sulfites, and metabisulfites. (2) The aqueous liquid preparation according to claim (1), which has a pH of 6 to 10.