JPH11269095A - Preparation containing hyaluronic acid - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an aqueous preparation containing hyaluronic acid or its salt and benzalkonium chloride, capable of efficiently improving a filtration process using a filter having a pore diameter of 0.22 μm as an effective sterilization process and substantially enabling the employment of the filter having the pore diameter of 0.22 μm. SOLUTION: This aqueous preparation containing hyaluronic acid is obtained by adding another surfactant, preferably a nonionic surfactant, to a system containing the hyaluronic acid, a hyaluronic acid salt or both the compounds as a main component and further benzalkonium chloride. Therein, the concentration of the benzalkonium chloride is 0.001-0.01 W/V%, and the concentration of the other surfactant is 0.001-1.0 W/V%.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to the sterile filtration of an aqueous preparation containing hyaluronic acid, a hyaluronic acid salt and both of them (hereinafter abbreviated as hyaluronic acids) and benzalkonium chloride. The present invention relates to an aqueous preparation which has been reduced and the production efficiency of the preparation has been dramatically improved.

[0002]

2. Description of the Related Art Benzalkonium chloride is a quaternary ammonium base type surfactant described in the Japanese Pharmacopoeia, and is generally used as a preservative for various aqueous preparations including pharmaceuticals such as eye drops and nasal drops. It has been used frequently. Since it has excellent antibacterial activity, it has a strong antiseptic effect against bacteria and fungi even at very low concentrations. Benzalkonium chloride is a mixture of fatty acids of a side chain alkyl group having 8 to 18 carbon atoms because fatty acids of natural fats and oils are used as a raw material.
And 14 are listed.

However, when benzalkonium chloride is added as a preservative in an aqueous preparation containing a hyaluronic acid as a main component at a normal concentration, it causes a change in the composition such as white turbidity and has been conventionally contraindicated. Techniques to avoid this contraindication include adding, for example, cyclodextrins to a system containing hyaluronic acid and benzalkonium chloride (Japanese Patent Laid-Open No. 1-16728), or using carbon in the side chain alkyl group of benzalkonium chloride to be used. Techniques such as limiting the number to 12 (JP-A-1-246227 and JP-A-2-148824) are disclosed.

[0004] These methods for avoiding contraindications are excellent in obtaining a clear preparation, but the judgment has been made by the naked eye. Even when using such a clear product, the sterile filterability, which is indispensable in the production of a sterile product, was examined.Compared with the case where hyaluronic acids and benzalkonium chloride were present together, the case where the product was used alone was compared. As a result, a decrease in filtration efficiency was observed. In other words, there is a possibility that a composition change that cannot be observed with the naked eye has occurred,
It has been found that even the use of the above-mentioned technology has not solved the problem in the production process of a preparation in which hyaluronic acids and benzalkonium chloride coexist.

[0005]

When benzalkonium chloride having 12 carbon atoms in the side chain alkyl group is used, filtration using a filter having a pore size of 0.22 μm, which guarantees sterility, is difficult. Efficiency is poor, and operability issues remain, such as frequently changing filters. In addition, the hole diameter is 0.22 μm
When it is difficult to use a filter having a pore size of 0.45 μm, it is theoretically conceivable to use a continuous multi-stage filter (two to three stages). . The present inventors have conducted intensive studies on reduction and improvement of the resistance to perfusion in a sterile filtration step when producing an aqueous preparation containing hyaluronic acids and using benzalkonium chloride as a preservative.

[0006]

An object of the present invention is to solve the above-mentioned problems, and the constitution of the present invention is based on a system containing hyaluronic acid, a hyaluronic acid salt or both as a main component and containing benzalkonium chloride, Another surfactant, preferably a nonionic surfactant, is blended, and the concentration of benzalkonium chloride is 0.001 to 0.01 W / V%, and the concentration of the other surfactant is 0.001 to 1.0 W. / V%.

That is, the present invention relates to an aqueous preparation mainly containing hyaluronic acid containing benzalkonium chloride as a preservative, and to which a small amount of another surfactant, preferably a nonionic surfactant or an amphoteric surfactant is added. This not only prevents a change in the composition, but also significantly improves the filtration resistance in a filtration membrane having a pore size of 0.22 μm during the aseptic filtration step.

When hyaluronic acids are used as raw materials for the preparation of pharmaceuticals, quasi-drugs or cosmetics,
Those having a molecular weight of about 300 to 2,000,000 are usually used. The larger the molecular weight, the more easily the blending with a cationic surfactant such as benzalkonium chloride occurs. In the present invention, a nonionic surfactant such as polyoxyethylene (20) sorbitan monooleate or an amphoteric surfactant such as betaine lauryl dimethylaminoacetate is added thereto. As a result, not only is an aqueous preparation excellent in clarity obtained, but also in the case of further sterile filtration, the resistance to the filter, which is a problem in the production process, is almost eliminated, and the conventional filter is poor in filtration and practically difficult to use. It is now possible to use a filter having a pore size of 0.22 μm.
According to the present invention, for example, even when the molecular weight of the hyaluronic acid used is 2,000,000 or more, it is possible to prevent a change in blending with benzalkonium chloride without affecting the preservative effect, and it is also possible to obtain the present invention. It is possible to perform sterile filtration using a filter having a pore size of 0.22 μm of the aqueous preparation.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION The hyaluronic acids used in the present invention may be those obtained by extracting from animal tissues such as chicken cockscomb, or those produced by fermentation engineering by culturing microorganisms or the like. . It may be a free form or a metal salt such as an alkali metal or an alkaline earth, and is used at a concentration of 0.01 to 1.0 W / V%.

As the benzalkonium chloride used as a preservative in an aqueous preparation, an alkyl group having 8 to 18 carbon atoms can be used. Its concentration is from 0.001 to 0.
01 W / V% is common. According to the present invention, a colorless and clear aqueous preparation which is stable over time can be obtained without causing a change in the composition at any benzalkonium chloride and concentration within this range.

[0011] side chain alkyl group as the C ₁₂ carbon atoms containing benzalkonium chloride 12 with relatively high ratio of - and benzalkonium chloride, carbon atoms of the side chain alkyl group is 1
The one containing benzalkonium chloride at a somewhat high ratio is referred to as C ₁₄ -benzalkonium chloride. C ₁₂ - When using benzalkonium chloride, in that the antimicrobial activity, C ₁₄ - poor preservative effect compared to benzalkonium chloride, sufficient preservative is the use of low concentration levels used in the aqueous formulation There is a concern that the effect cannot be obtained. Suppose C
Even when preparing a hyaluronic acid-containing aqueous preparation using _12- benzalkonium chloride, for example, at the time of aseptic filtration essential for the production process of eye drops, a pore size of 0.22 μm excellent in sterilization performance is obtained. Passing through a filter is difficult in the manufacturing process, and it is necessary to prepare a sterile preparation using a filter having a larger pore size. However, in the present invention, for example the hyaluronic acid In preparing the eye drop containing 0.1 W / V% is what carbon number ratio (C ₁₂ body, the abundance ratio of C ₁₄ body or the like) a mixture of Even when used, or when a plurality of raw materials having different ratios are used together, the total benzalkonium chloride concentration is 0.00
1 to 0.01 W / V% range, preferably 0.002 to
At any concentration within the range of 0.006 W / V%, filtration using a filter having a pore size of 0.22 μm is possible.

In the present invention, a nonionic surfactant or an amphoteric surfactant is used as another surfactant. Examples of the nonionic surfactant include polyoxyethylene sorbitan fatty acid esters, polyoxyethylene hydrogenated castor oil, polyoxyethylene castor oil, polyoxyethylene alkyl ether, polyoxyethylene polyoxypropylene ether, and the like. .

Representative examples of polyoxyethylene sorbitan fatty acid esters include polyoxyethylene (20) sorbitan monooleate, polyoxyethylene (20) sorbitan monostearate, and polyoxyethylene (20) sorbitan monopalmitate. Representative examples of polyoxyethylene hydrogenated castor oil include polyoxyethylene (60) hydrogenated castor oil, polyoxyethylene (40) hydrogenated castor oil, and the like. Representative examples of polyoxyethylene castor oil include polyoxyethylene (35) castor oil.

A typical example of the polyoxyethylene alkyl ether is polyoxyethylene (9) lauryl ether. Typical examples of polyoxyethylene polyoxypropylene ether include polyoxyethylene (30) polyoxypropylene (6) decyltetradecyl ether and the like. These nonionic surfactants may be used alone or in combination of a plurality of types. The concentration of the nonionic surfactant used is 0.001 to 1.0 W / V%, preferably 0.0
It is 0.5 to 0.1 W / V%, and more preferably 2 to 10 parts by weight based on 1 part by weight of benzalkonium chloride contained.

Examples of the amphoteric surfactant include betaines, aminocarboxylates, and imidazoline derivatives. Examples of the betaine type include carboxy betaine type such as betaine lauryl dimethylaminoacetate, and amino carboxylate. These amphoteric surfactants may be used alone or in combination of a plurality of types. The concentration of the amphoteric surfactant used is 0.001 to 1.0 W / V%, preferably 0.005 to 0.1 W / V%. Furthermore, the preservative effect can be further enhanced by adding a preservative effect enhancer such as edetate to the colorless and clear aqueous preparation obtained by the present invention.

[0016]

EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples, but the present invention is not limited to these examples.

Example 1 Sodium hyaluronate (average molecular weight: 900,000) 0.1 g Benzalkonium chloride 0.005 g Sodium chloride 0.9 g Polyoxyethylene (20) sorbitan monooleate 0.02 g A total amount of 100 ml was added by adding sterile purified water. Preparation method Sodium chloride, benzalkonium chloride in sterile purified water,
After dissolving polyoxyethylene (20) sorbitan monooleate, sodium hyaluronate was added and dissolved therein. The total volume was adjusted to 100 ml by adding sterilized purified water, and the pH was adjusted to 6.5 using dilute hydrochloric acid or sodium hydroxide.

Example 2 Sodium hyaluronate (average molecular weight: 900,000) 0.1 g Benzalkonium chloride 0.03 g Sodium edetate 0.01 g Sodium chloride 0.9 g Polyoxyethylene (20) sorbitan monooleate 0.01 g Sterilization and purification sodium chloride to a total volume of 100ml preparation process sterile purified water appropriate amount by adding water, benzalkonium chloride, sodium edetate, polyoxyethylene (20) was dissolved sorbitan monooleate, and this was dissolved by adding sodium hyaluronate . The total volume was adjusted to 100 ml by adding sterilized purified water, and the pH was adjusted to 6.5 using dilute hydrochloric acid or sodium hydroxide.

Example 3 Sodium hyaluronate (average molecular weight: 2,000,000) 0.1 g Benzalkonium chloride 0.005 g Sodium chloride 0.9 g Polyoxyethylene (20) sorbitan monooleate 0.02 g A total of 100 ml was prepared by adding sterile purified water. Method Sodium chloride, benzalkonium chloride, and polyoxyethylene (20) sorbitan monooleate were dissolved in an appropriate amount of sterilized purified water, and sodium hyaluronate was added to this and dissolved. Add sterilized purified water to make the total volume 100 ml,
The pH was adjusted to 6.5 using dilute hydrochloric acid or sodium hydroxide.

Example 4 Sodium hyaluronate (average molecular weight 2,000,000) 0.1 g Benzalkonium chloride 0.03 g Sodium edetate 0.01 g Sodium chloride 0.9 g Polyoxyethylene (20) sorbitan monooleate 0.01 g Sterilization and purification sodium chloride to a total volume of 100ml preparation process sterile purified water appropriate amount by adding water, benzalkonium chloride, sodium edetate, polyoxyethylene (20) was dissolved sorbitan monooleate, and this was dissolved by adding sodium hyaluronate . The total volume was adjusted to 100 ml by adding sterilized purified water, and the pH was adjusted to 6.5 using dilute hydrochloric acid or sodium hydroxide.

Example 5 Sodium hyaluronate (average molecular weight: 1,000,000) 0.1 g Benzalkonium chloride 0.03 g Sodium edetate 0.01 g Sodium chloride 0.9 g Betaine lauryl dimethylaminoacetate 0.05 g Add sterile purified water sodium chloride to a total volume of 100ml preparation process sterile purified water appropriate amount, benzalkonium chloride, sodium edetate were dissolved lauryl betaine, and this was dissolved by adding sodium hyaluronate. Add sterile purified water and add 100m
and the pH was adjusted to 6.5 using dilute hydrochloric acid or sodium hydroxide.

Comparative Examples 1 and 2 Polyoxyethylene (20) was used as a nonionic surfactant.
A hyaluronic acid preparation was produced in the same manner as in Example 1 except that sorbitan monooleate was not added. A hyaluronic acid preparation was produced in the same manner as in Example 2 except that polyoxyethylene (20) sorbitan monooleate was not added as a nonionic surfactant, and Comparative Example 2 was used.

Appearance stability test A hyaluronic acid preparation was prepared according to the formulations described in Examples 1 to 5 and Comparative Examples 1 and 2, and the appearance stability when stored at 40 ° C. and 75% RH for 6 months was evaluated. Observed. Example 1
5 and Comparative Examples 1 and 2 were all colorless and clear after 6 months.

Preservation Efficacy Test Hyaluronic acid preparations were prepared according to the formulations described in Examples 1 to 4, and the preservative effect of the preparations on bacteria and fungi was confirmed. The test was carried out in accordance with the thirteenth revised Japanese Pharmacopoeia Preservation Efficacy Test Method, and was evaluated by the residual ratio of the bacteria to the number of inoculated bacteria 14 and 28 days after inoculation. In addition, a sterile physiological saline solution was used as a control solution. The criterion for bacteria was 0.1% or less of the number of inoculated bacteria after 14 days and 1 day after 28 days.
As for fungi equivalent to or less than the level after 4 days, the number of inoculated bacteria is equal to or less than 14 days later and equal to or less than 28 days after inoculation.

As the bacterial species, Escherichia coli, Pseudo
monas aeruginosa, Staphylococcusaureus, Candida a
lbicans, Aspergillus niger, Examples 1-4
All products were "compatible" and all control products were "unsuitable".

Sterile Filtration Test A hyaluronic acid ophthalmic solution preparation was prepared according to the formulations described in Examples 1 to 5 and Comparative Examples 1 and 2, and this was used as a sample solution to compare the loper resistance. Purified water was used as a control solution.
In order to quantitatively evaluate the filtration performance, that is, the degree of clogging due to adsorption to the filter, measure the possible filtration amount (volume / time) of each sample solution within a certain time while measuring the flow rate at a certain temperature. And performed comparative evaluation. The filtration test conditions are shown below.

Filtration filter used under filtration conditions : Mirex-GS manufactured by Millipore
Type 0.22μm filter Pump used: Yamato Master Flex 7
518-02 type pump Sample solution temperature: 30 ° C Filter temperature: 30 ° C Flow rate setting: Set the flow rate to 20 ml per minute when filtering purified water Filter time: 5 minutes As a membrane filter Millipore My Rex-G
Using a Yamato pump equipped with an S-type 0.22 μm filter, each sample solution of Examples 1 to 5 and Comparative Examples 1 and 2 and purified water were accurately adjusted at a constant flow rate from the start of filtration. Was passed through a membrane filter for 5 minutes. During this time, the amount of filtration of the solution that could pass through the membrane filter was measured every minute, and the results were shown in Table 1.
It was shown to.

[0028]

[Table 1]

With respect to the formulations of Example 2, Example 4, Comparative Example 1 and Comparative Example 2, a change in the physical properties of the preparation was compared between the sample before filtration and the sample after filtration. The results are shown in Tables 2, 3, 4 and 5.

[0030]

[Table 2]

[0031]

[Table 3]

[0032]

[Table 4]

[0033]

[Table 5]

In any of the examples to which polyoxyethylene (20) sorbitan monooleate was added, no change in blending with benzalkonium chloride was observed over time. The preservative effect was not affected by the nonionic surfactant, and a sufficient preservative effect was obtained. Furthermore, it became clear that sterile filtration using a membrane filter having a pore size of 0.22 μm can be performed without any problem. On the other hand, in the comparative example, the filtration efficiency was poor, the hyaluronic acid concentration and the benzalkonium chloride concentration were changed, and sterile filtration with a 0.22 μm filter was substantially difficult.

[0035]

Industrial Applicability In an aqueous preparation containing a hyaluronic acid and benzalkonium chloride, the efficiency of sterilization by filtration is significantly improved by adding another surfactant, and a 0.22 μm filter having a pore size of 0.22 μm in the production process. Can be used.

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Toshihiko Umeda 3-5-1 Asahicho, Machida-shi, Tokyo Denki Kagaku Kogyo Co., Ltd. (72) Inventor Hiromichi Kitagawa 3 Asahicho, Machida-shi, Tokyo No.5-1, Denki Kagaku Kogyo Co., Ltd.

Claims (5)

[Claims] 1. An aqueous preparation characterized in that another surfactant is added to a system containing hyaluronic acid, a hyaluronic acid salt or both as a main component, and benzalkonium chloride. 2. The aqueous preparation according to claim 1, wherein the other surfactant is a nonionic surfactant. 3. A benzalkonium chloride concentration of 0.00
1 to 0.01 W / V%, the concentration of other surfactant is 0.0
The aqueous preparation according to claim 1, wherein the aqueous preparation has a concentration of 01 to 1.0 W / V%. 4. The aqueous preparation according to claim 1, wherein the aqueous preparation is an eye drop. 5. A system containing hyaluronic acid, hyaluronic acid salt or both as a main component and benzalkonium chloride, mixed with another surfactant, and filtered through a sterile filter membrane having a pore size of 0.22 μm. A method for producing an aqueous preparation, characterized in that: