JPH10130156A - Eye drop composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an eye drop composition capable of alleviating fear of adverse effects by an antiseptic, maintaining antimicrobial properties by adding borate ion, especially effective against microorganisms resistant to benzalkonium chloride, by adding a very small amount of an antiseptic to an eye drop composition. SOLUTION: This eye drop composition contains 0.001-0.005wt.% benzalkonium chloride and 8.14mM-0.244M borate ion.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention reduces the risk of side effects of a preservative by adding a small amount of a preservative, and maintains antibacterial properties by adding borate ions. The present invention relates to eye drops that are effective against bacteria.

[0002]

2. Description of the Related Art Since eye drops are pharmaceutical compositions containing water as a medium, it has been considered that the addition of a bactericidal preservative is an essential element in preserving the eye drops. According to recent research reports, there is a report indicating that the combination of preservatives may cause keratoconjunctival disorder and corneal epithelial disorder (New Ophthalmology 8 (10): 1599-160).
3, 1991; Ophthalmology 33: 533-538 (199
1), Toxicology and Applied
Pharmacology 62, 251 161 (1
982)). It is natural that the eye drops have antibacterial properties as a pharmaceutical preparation, but the amount of preservatives is reduced to prevent the above-mentioned fear of the occurrence of the trouble, and the antibacterial properties of the eye drops are maintained. It is desired. Tokuhei 07-54
No. 56 discloses an ophthalmic solution containing benzalkonium chloride and p-hydroxybenzoate as a preservative and further containing a chelating agent, which has a low concentration of the preservative, has a bactericidal effect, and is less irritating. .

[0003]

However, Japanese Patent Publication No. 07-5456 discloses that the amount of benzalkonium chloride having a strong bactericidal effect is reduced, paraoxybenzoic acid ester having a slightly weaker antibacterial activity is added, and the bactericidal effect is further reduced. It merely discloses those that were physically preserved by the addition of certain sodium edetate. The present invention reduces the danger by reducing the concentration of benzalkonium chloride, which is a preservative that has a strong bactericidal effect and may cause eye damage, and maintains the antibacterial property, and furthermore, has an antibacterial property. We also succeeded in obtaining a strong bactericidal effect against benzalkonium chloride-resistant bacteria, which is a problem.

[0004]

The ophthalmic solution composition of the present invention is characterized by containing the following components (a) and (b). (A) 0.001 to 0.005% by weight of benzalkonium chloride. (B) Borate ions at a concentration of 8.14 mM to 0.244 M.

[0005]

BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, benzalkonium chloride as the component (a) is blended as a preservative. Benzalkonium chloride is the most commonly used preservative in eye drops, but its drawbacks include the presence of resistant bacteria, and its high concentration can cause eye damage and irritation. . Therefore, in the present invention, the amount of benzalkonium chloride to be added to the eye drop composition is 0.001 to 0.005% by weight, preferably 0.002%, from the viewpoints of ensuring the preservative effect and preventing ocular damage and irritation. It was set to a very small range of 0.003% by weight. And, in the present invention, even in the trace amount range of benzalkonium chloride, the antibacterial action, particularly the antibacterial action against benzalkonium chloride-resistant bacteria can be improved by adding an appropriate amount of borate ion as the component (b). I found it.

[0006] Borate ions are used in eye drops in borate buffers and the like, and are known to have an antibacterial effect. It is also known that when the borate ion concentration exceeds 0.244M, allergies may occur. However, the combination of the borate buffer and low concentration of benzalkonium chloride, as compared with other buffers, for example, a combination of a citrate buffer, a phosphate buffer, and an acetate buffer, as in Experimental Example 1 described below, It is clearly excellent in antibacterial properties, and exerts its effects irrespective of the drug components to be blended, as in Experimental Example 2 also described later. Moreover, as a result of the study, it was found that the combination of borate ion and benzalkonium chloride used was effective within a certain range of the compounding amount.

[0007] The borate ion of the component (b) is contained in the ophthalmic solution of the present invention in the range of 8.14 mM (mmol) to 0.244.
M (mol), preferably in the range of 10 mM to 0.2 M. This compounding amount is 8.14 mM
If it is less than 0.1, the antibacterial effect of the present invention cannot be obtained.
If it exceeds 244M, there is a concern that allergy will occur. Borate ions are supplied from a borate such as boric acid or borax, and a borate buffer is formed by combining boric acid / borate.

[0008] The eye drop composition of the present invention comprises the above (a),
(B) In addition to both essential components, various optional components can be blended. For example, drugs such as timolol maleate, azelastine hydrochloride, sodium hyaluronate, polysorbate 80, polyoxyl stearate, and polyoxyethylene hydrogenated castor oil derivative And other nonionic surfactants can be used. In addition, polyhydric alcohols such as ethanol, propylene glycol, glycerin, and polyethylene glycol as solvents, and sodium edetate as a chelating agent can be blended. The eye drop composition of the present invention preferably has a pH of about 5 to 8. The osmotic pressure ratio may be generally adjusted to be 1 and can be adjusted with a polyhydric alcohol such as sodium chloride, potassium chloride or glycerin.

[0009]

According to the present invention, benzalkonium chloride as a preservative is blended in a low concentration range where there is no risk of causing irritation or eye damage, and a specific amount of borate ion is added thereto. It can enhance antibacterial properties and exhibits an excellent effect especially on benzalkonium chloride resistant bacteria.

[0010]

【Example】

Example 1 0.004 parts of concentrated benzalkonium chloride solution 50 was added to 90 parts of sterilized purified water, and 0.005 part of sodium edetate was added.
1.5 parts of boric acid (0.243 M), 0.019 parts of borax (0.49 mM) and 0.13 parts of sodium chloride were added in sequence, stirred and dissolved, and sterilized purified water was added in an appropriate amount to 100 ml.
And This solution is filtered through a 0.22 μm membrane filter to obtain the eye drop composition of the present invention. In addition,
"Parts" are by weight unless otherwise specified. Also,
"Concentrated benzalkonium chloride solution 50" indicates a 50-55% solution of benzalkonium chloride. The content of benzalkonium chloride in the eye drop composition is 0.002%.
The borate ion concentration of the eye drop composition of Example 1 was 0.24.
3M.

Example 2 0.004 parts of concentrated benzalkonium chloride solution 50 was added to 90 parts of sterilized purified water, and 1.5 parts of boric acid (0.243M) was added.
0.015 parts of borax (0.39 mM) and 0.13 parts of sodium chloride are sequentially added, stirred and dissolved, and sterilized purified water is added to make up to 100 ml. This solution is filtered through a 0.22 μm membrane filter to obtain the eye drop composition of the present invention. The borate ion concentration of the eye drop composition of Example 2 is 0.243M.

Example 3 0.004 parts of concentrated benzalkonium chloride solution 50 was added to 90 parts of sterile purified water, and 0.005 part of sodium edetate was added.
1.5 parts of boric acid (0.243 M), 0.14 parts of borax (3.67 mM) and 0.12 parts of sodium chloride are sequentially added, stirred and dissolved, and an appropriate amount of sterilized purified water is added to make 100 ml. This solution is filtered through a 0.22 μm membrane filter to obtain the eye drop composition of the present invention. The borate ion concentration of the eye drop composition of Example 3 is 0.243M.

Example 4 0.004 parts of concentrated benzalkonium chloride solution 50 was added to 90 parts of sterilized purified water, and 0.005 part of sodium edetate was added.
Boric acid 0.05 parts (0.008M), borax 0.002
Parts (0.052 mM) and 0.875 parts of sodium chloride were added in sequence, stirred and dissolved, and an appropriate amount of sterilized purified water was added thereto to obtain 100 parts.
ml. This solution is filtered through a 0.22 μm membrane filter to obtain the eye drop composition of the present invention. The borate ion concentration of the eye drop composition of Example 4 was 0.008 M
It is.

Example 5 0.004 parts of concentrated benzalkonium chloride solution 50 was added to 90 parts of sterilized purified water, and 0.005 part of sodium edetate was added.
0.5 part (0.081 M) of boric acid, 0.005 part (0.131 mM) of borax and 0.67 part of sodium chloride are sequentially added, stirred and dissolved, and an appropriate amount of sterilized purified water is added to make 100 ml. This solution is filtered through a 0.22 μm membrane filter to obtain an eye drop composition of the present invention. Example 5
Has a borate ion concentration of 0.081M.

EXAMPLE 6 0.004 parts of concentrated benzalkonium chloride solution 50 was added to 90 parts of sterile purified water, and 0.005 part of sodium edetate was added.
One part of boric acid (0.162M), 0.009 part of borax (0.236mM) and 0.3 part of sodium chloride are sequentially added, stirred, dissolved, and an appropriate amount of sterilized purified water is added to 100ml.
And This solution is filtered through a 0.22 μm membrane filter to obtain an eye drop composition of the present invention. The borate ion concentration of the eye drop composition of Example 6 is 0.162M.

Comparative Example 1 0.004 parts of concentrated benzalkonium chloride solution 50 was added to 90 parts of sterile purified water, and 0.005 part of sodium edetate was added.
0.3 parts of sodium citrate, 0.023 parts of citric acid,
0.82 parts of sodium chloride are sequentially added, stirred and dissolved, and sterilized purified water is added to make up to 100 ml. 0.22
The solution is filtered through a μm membrane filter to obtain an ophthalmic solution composition of Comparative Example.

Comparative Example 2 0.004 parts of concentrated benzalkonium chloride solution 50 was added to 90 parts of sterilized purified water, and 0.005 part of sodium edetate was added.
0.3 parts of sodium acetate, 0.045 parts of glacial acetic acid and 0.77 parts of sodium chloride are sequentially added, stirred and dissolved, and sterilized purified water is added to make 100 ml. This solution was filtered through a 0.22 μm membrane filter to obtain an ophthalmic solution composition of Comparative Example.

Comparative Example 3 0.004 parts of concentrated benzalkonium chloride solution 50 was added to 90 parts of sterilized purified water, and 0.005 part of sodium edetate was added.
1.3 parts of sodium hydrogen phosphate, 0.8 parts of disodium hydrogen phosphate and 0.1 part of sodium chloride are sequentially added and stirred,
Dissolve and add sterile purified water to make 100 ml. This solution was filtered through a 0.22 μm membrane filter to obtain an ophthalmic solution composition of Comparative Example.

Example 7 0.00342 parts of timolol maleate and 0.0
04 parts of concentrated benzalkonium chloride solution 50 was sterilized with purified water 9
In addition to 0 parts, sodium edetate 0.005 part, boric acid 0.5 part (0.081M), borax 0.0015 part (0.039 mM) and sodium chloride 0.64 part are sequentially added, stirred and dissolved. , Add an appropriate amount of sterile purified water and add 100ml
And This solution is filtered through a 0.22 μm membrane filter to obtain an eye drop composition of the present invention. The borate ion concentration of the eye drop composition of Example 7 is 0.081M.

Example 8 0.025 parts of azelastine hydrochloride and 0.004 parts of concentrated benzalkonium chloride solution 50 were added to 90 parts of sterilized purified water, and 0.01 parts of sodium edetate and 1.5 parts of boric acid ( 0.243M), borax 0.049 parts (0.001
M) and 0.1 part of sodium chloride were sequentially added, stirred and dissolved, and an appropriate amount of sterilized purified water was added to make 100 ml. This solution was filtered through a 0.22μ membrane filter,
The ophthalmic composition of the present invention is provided. The borate ion concentration of the eye drop composition of Example 8 is 0.244M.

[Experimental Examples] In the following experimental examples, the antibacterial effect was evaluated by the following method. (1) Preparation of Inoculated Bacterial Solution Bacteria: Using SCD agar slant medium, transplantation is repeated daily at least 3 days before at 37 ° C., and it is confirmed that the cells have grown sufficiently. If sufficient growth is not observed, the subculture is repeated until sufficient growth is observed. S the day before use
One platinum loop of the bacterium is transferred from a CD agar medium to 30 ml of an SCD medium, and cultured at 37 ° C. for 16 to 18 hours. After culture
Put the cultured SCD medium in a sterile centrifuge tube, and
Centrifuge at pm for 5 minutes. The supernatant was removed by decantation, and 30 ml of sterile PBS was used under the same conditions.
Wash twice. Using a bacterial counting plate, the suspension is diluted with sterile PBS to prepare a suspension having a bacterial concentration of 4 × 10 ⁶ to 2 × 10 ⁷ cells / ml, which is used as an inoculum. Further, the number of viable cells is measured by an agar plate pour method using an SCD agar medium.

Fungi: 1) Candida albica
The cells are cultured and prepared at 25 ° C. using ns GP agar slant medium in the same manner as bacteria. 2) Aspergillus niger, Asper
culturing at 25 ° C. for at least 7 days using a Gillus pullulans GP agar slant medium,
The conidia grown on the medium are suspended in 30 ml of sterile PBS and filtered with sterilized gauze. The filtrate is taken into a sterile centrifuge tube and separated at 3000 rpm for 5 minutes. The supernatant was removed by decantation, and 30 ml of sterile PBS was used.
Wash twice under the same conditions. Sterile PB using bacteria calculator
After dilution with S, a suspension having a bacterial concentration of 4 × 10 ⁶ to 2 × 10 ⁷ cells / ml is prepared and used as an inoculum. Further, the number of viable cells is measured by agar plate pour method using a GP agar medium.

(2) Separation of Test Material 19.5 ml of the test sample is dispensed into a sterilized test tube or a suitable container. PBS is used as a control. All test samples and controls are filtered using a 0.22 μm membrane filter.

(3) Inoculation of bacteria 0.5 ml of the inoculated bacterial solution was added to 19.5 ml of the test sample and control.
, And mixed well to obtain a bacterial inoculum sample (test solution).
The concentration (number / ml) in 20 ml of this test solution is defined as the number of inoculated bacteria. For example, in the case of 2.0 × 10 ⁷ inoculum / ml,
The number of inoculated bacteria is 5.0 × 10 ⁵ cells / ml.

(4) Storage and Sampling of Bacterial Inoculation Sample The test solution inoculated with the bacteria was stored at 30 to 32 ° C, and the test solution inoculated with the fungus was stored at 20 to 25 ° C.
After 6 hours, 1, 4 and 7 days, the mixture is thoroughly stirred or mixed, 1 ml of the test solution is aseptically sampled, and the time-dependent decrease in the number of viable cells is measured by an agar plate pour method.

(5) Measurement of viable cell count: Agar plate pour method. Preparation of agar plates Use SCD agar for bacteria and GP agar for fungi. 1 ml of the test solution is suspended in 9 ml of the LP diluent to prepare a 10-fold diluted sample. The same operation is repeated using 1 ml of the 10-fold diluted solution to prepare a 10-fold serial diluted solution of the sample. Place 1 ml of each serial dilution in two sterile dishes each. Then, 15 to 20 ml of a predetermined agar medium sterilized and dissolved in advance and kept at about 50 ° is aseptically poured,
Mix the sample and the medium well, taking care not to adhere to the lid of the petri dish, and cool and solidify.

(6) Culture The plate is turned upside down and cultured. For bacteria (SCD agar), culture at 30 ° C. for 3 days. For fungi (GP agar medium), culture at 25 ° C. for 7 days. (7) Calculation of the number of colonies Count the number of colonies that occurred immediately after culturing. The number of villages is calculated by selecting a flat plate having no diffused villages and having 20 to 200 settlements on one flat plate. (8) Calculation of the number of bacteria The number of viable bacteria in 1 ml of the test solution is measured from the number of colonies (average value) obtained per plate and the dilution factor.

(9) Calculation of Residual Rate The residual rate is expressed as a percentage with respect to the number of inoculated bacteria by the following equation 1.

(Equation 1)

(10) Indicator strain used: Pseudomonas cepacia: IFO14
595 Pseudomonas cepacia resistance: IFO
15124 Candida albicans: IFO 1594 Aspergillus niger: IFO 4414 Aspergillus pullulans: IFO
5353 (11) Determination Bacteria (Ps. Cepasia, resistant Ps. Cepaci)
a): 0.1% or less at 6 hours, negative at 24 hours, 7
Fungi that are negative until the day are considered fungi (Candida albicans, Asper)
gillus niger, Aspergillus
pullulans): 0.01% or less in 7 days, after which no growth is accepted

Experimental Example 1 The eye drops of Example 1 and Comparative Examples 1 to 3 were prepared using Ps.
cepacia, resistant Ps. The antibacterial effect on C. pacifica was confirmed and is shown in Tables 1 and 2 below. While the ophthalmic solution of Example 1 containing borate ion and containing borate buffer showed an excellent antibacterial effect, Comparative Examples 1 to 3 using a citrate buffer, a phosphate buffer and an acetate buffer all showed an excellent antibacterial effect. No antibacterial properties were observed.

[0031]

Table 1: Experimental example 1 (residual rate:%) Ps. Sepacia aging Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 0 0 hours 7.6 65 12 43 6 hours 0 44 2.6 2.7 1 day 0 129 1.5 25 4 days 0 106 1.8 65 7 days 0 171 38 153

[0032]

Table 2: Experimental example 1 (residual rate:%) Ps. Cepacia resistance aging Example 1 Comparative example 1 Comparative example 2 Comparative example 0 0 0.067 27 3.3 326 6 hours 0 17 1.9 52 1 day 0 133 5.6 253 4 days 4 173 12 267 7 days 0 160 95 147

EXPERIMENTAL EXAMPLE 2 The antibacterial effect of the eye drops of the present invention [(timolol maleate) of Example 7 and azelastine hydrochloride) containing different drugs was confirmed. The results are shown in Table 3. Was. It turns out that the ophthalmic solution of the present invention exerts an antibacterial effect without depending on the combination drug.

[0034]

Table 3: Experimental example 2 (residual rate:%) Ps. Cepacia Ps. Cepacia (resistance) with time Example 7 Example 8 Example 7 Example 8 0 hour 29 18 52 41 6 hours 0 0 0.01 0.01 1 day 0 0 0 0 4 days 0 0 0 0 7 days 0 0 0 0

Experimental Example 3 The antibacterial activity of the eye drops of the present invention was confirmed for various indicator strains by changing the borate ion concentration. It can be seen that excellent antibacterial activity is exhibited at a borate ion concentration in the range of 8.14 mM to 0.244 M.

[0036]

[Table 4]Table 4: Experimental example 3 (residual rate:%) Ps. cepacia Elapsed time Example 1 Example 2 Example 3 Example 4 Example 4 Example 5 Example 6 0 hours 7.6 0.16 0.026 53 20.1 13.4 6 hours 0 0 0 0.058 0 0 1 day 0 0 0 0 0 0 4 days 0 0 0 0 0 0 7th 0 0 0 0 0 0

[0037]

[Table 5]Table 5: Experimental example 3 (residual rate:%) Ps. cepacia resistance Elapsed time Example 1 Example 2 Example 3 Example 4 Example 4 Example 5 Example 6 0 hour 0.067 0.036 0.01 50 53.4 3.3 6 hours 0 0 0 0.04 0.02 0.01 1 day 0 0 0 0 0 0 4 days 0 0 0 0 0 0 7th 0 0 0 0 0 0

[0038]

[Table 6]Table 6: Experimental example 3 (residual rate:%) Candida albicans Elapsed time Example 1 Example 2 Example 3 Example 4 Example 4 Example 5 Example 6 0 hours 0.17 0 0 34 20.4 2.1 6 hours 0 0 0 0 0 0 1 day 0 0 0 0 0 0 4 days 0 0 0 0 0 0 7th 0 0 0 0 0 0

[0039]

[Table 7]Table 7: Experimental example 3 (residual rate:%) Aspergillus niger Elapsed time Example 1 Example 2 Example 3 Example 4 Example 4 Example 5 Example 6 0 hour 10 0 0 105 84.1 20.3 6 hours 0 0 0 0.24 0.15 0.02 1 day 0 0 0 0.017 0 0 4 days 0 0 0 0.014 0 0 7th 0 0 0 0.005 0 0

[0040]

[Table 8]Table 8: Experimental example 3 (residual rate:%) Aspergillus pullulans Elapsed time Example 1 Example 2 Example 3 Example 4 Example 4 Example 5 Example 6 0 hour 0 0.03 0.01 17 3.2 0.05 6 hours 0 0 0 0 0 0 1 day 0 0 0 0 0 0 4 days 0 0 0 0 0 0 7th 0 0 0 0 0 0

Claims (1)

[Claims] 1. a) 0.001 to 0.005% by weight of benzalkonium chloride, and (b) 8.14 mM.
An eye drop composition comprising a borate ion at a concentration of 0.244M.