HK1190089B - Ophthalmic solution containing hyaluronic acid or salt thereof and propylene glycol - Google Patents

Abstract

A water-based ophthalmic solution comprising hyaluronic acid or a salt thereof at a concentration of 0.03-0.5% (w/v) and propylene glycol at a concentration of 0.1-1.0% (w/v), wherein benzalkonium chloride is contained at a concentration of 0.001-0.002% (w/v) as a sole preservative agent and an ionic tonicity agent is also contained at such a concentration that the osmotic pressure ratio of the ophthalmic solution becomes 0.9-1.1.

Description

Eye drops containing hyaluronic acid or its salt and propylene glycol

Technical Field

The present invention relates to an aqueous eye drop containing hyaluronic acid or a salt thereof at a concentration of 0.03 to 0.5% (w/v) and propylene glycol at a concentration of 0.1 to 1.0% (w/v), the eye drop containing benzalkonium chloride at a concentration of 0.001 to 0.002% (w/v) as a sole preservative and containing a concentrated solution having an osmotic pressure ratio of 0.9 to 1.1A moderately ionic isotonic agent. The present invention also relates to a method for improving the storage efficiency of an eye drop containing sodium hyaluronate in a concentration of 0.1 or 0.3% (w/v) and benzalkonium chloride as the only preservative, and for achieving a kinematic viscosity of 3.0 to 4.0 or 17 to 30mm²(iii)/s and preventing dripping (drip) of the eye drop, the method comprising: a step (a) of mixing sodium hyaluronate in an amount to achieve a concentration of 0.1 or 0.3% (w/v) in the eye drop, benzalkonium chloride in an amount to achieve a concentration of 0.001-0.002% (w/v) in the eye drop, propylene glycol in an amount to achieve a concentration of 0.1-1.0% (w/v) in the eye drop, and an ionic isotonic agent in an amount to achieve an osmotic pressure ratio of 0.9-1.1 in the eye drop; a step (b) of adjusting the pH of the eye drop to 6.0 to 7.0; and a step (c) of filling the eye drop into an eye drop container.

Background

Dry eye is a disease that progresses from symptoms of an unpleasant degree such as dry eyes and a feeling of rubbing with foreign bodies, and if worsened, it causes a serious obstacle to daily life. The number of dry eye patients increases year by year with the advent of aging society and the increase in VDT (video display terminal) operations such as personal computers, and it is estimated that the number of patients in the united states is 1000 or more thousands, and it is also said that the number of patients in japan is 800 or more thousands.

Although the pathology of dry eye has not been fully elucidated, dry eye is known to cause corneal conjunctival epithelial damage, eventually leading to abnormal vision. Therefore, it is extremely important to treat the corneal conjunctival epithelium damage caused by dry eye early and appropriately.

At present, eye drop treatment is the most common treatment method for dry eye, and in japan, eye drops containing sodium hyaluronate are generally used for dry eye treatment.

There are eye drops containing sodium hyaluronate (hereinafter also referred to simply as "sodium hyaluronate eye drops") forMultidose type (Hyalein) capable of being used repeatedly and freely opening and re-closing cap^{(registered trademark)}0.1% of eye drops and Hyalein^{(registered trademark)}Eye drops 0.3%, etc.) and unit dose forms for single use (Hyalein)^{(registered trademark)}Mini eye drops 0.1%, Hyalein^{(registered trademark)}Mini eye drops 0.3%, etc.).

In the multi-dose type sodium hyaluronate ophthalmic solution, benzalkonium chloride is added as a preservative because the cap is opened and resealed, whereas the unit dose type is used once (used up), and therefore, the preservative is not added.

Preservatives such as benzalkonium chloride are pointed out to be at risk of inducing corneal damage, and benzalkonium chloride, for example, as disclosed in clinicaland experimental ophthalmology,32,180-184 (2004) (non-patent document 1), is known to cause damage to the corneal epithelium depending on the concentration. As described above, since dry eye is originally a disease accompanied by damage to corneal epithelium, sodium hyaluronate eye drops of a single dose type are used for severe dry eye patients. On the other hand, it is difficult to prescribe a single-dose type of sodium hyaluronate ophthalmic solution to all dry eye patients due to problems such as production cost, and therefore it is also conceivable to reduce the risk of occurrence of corneal epithelial injury by reducing the concentration of preservative in a multi-dose type of ophthalmic solution. However, in practice, if the concentration of the preservative in the eye drops is reduced, sufficient storage efficiency cannot be obtained when the eye drops are used in a multi-dose form, and the physical and chemical properties of the eye drops may be different from those of conventional sodium hyaluronate eye drops because the compounding ingredients change with the reduction of the preservative.

Further, although eye drops containing sodium hyaluronate, benzalkonium chloride and propylene glycol are disclosed in japanese patent laid-open nos. 2004-359629 (patent document 1), 2009-196903 (patent document 2) and 2009-161454 (patent document 3), there is no description at all on the point that the benzalkonium chloride concentration in the eye drops is 0.0025% or less, but the eye drops have a sufficient storage effect and have the same physicochemical properties as those of conventional sodium hyaluronate eye drops.

Patent document 1: japanese patent laid-open publication No. 2004-359629

Patent document 2: japanese patent laid-open No. 2009-196903

Patent document 3: japanese patent laid-open No. 2009-161454

Non-patent document 1: ClinicalandExperimentOphthematology, 32, 180-

Disclosure of Invention

As described above, it is an interesting problem to search for an eye drop which has a sufficient storage efficiency even when the benzalkonium chloride concentration is reduced and has the same physicochemical properties as those of conventional sodium hyaluronate eye drops.

The present inventors have found that sodium hyaluronate ophthalmic solution containing 0.002% (w/v) benzalkonium chloride cannot achieve sufficient storage efficiency, but that sufficient storage efficiency can be achieved even when benzalkonium chloride concentration is reduced to 0.001% (w/v) when propylene glycol is added.

On the other hand, the inventors found that if the propylene glycol added to the sodium hyaluronate eye drops exceeds 1% (w/v), the content of the propylene glycol exceeds "Hyalein 11 months (revised 7 th edition) in 2010^{(registered trademark)}0.1% of eye drops and Hyalein^{(registered trademark)}0.3 percent of eye drops and Hyalein^{(registered trademark)}Mini eye drops 0.1%, Hyalein^{(registered trademark)}The present inventors have completed the present invention by setting the upper limit of the amount of propylene glycol to 1% (w/v) because the allowable range of the kinematic viscosity and/or osmotic pressure ratio of sodium hyaluronate eye drops described in the mini eye drops 0.3% pharmaceutical summary (hereinafter also referred to as "Hyalein summary") "is defined.

Further, the present inventors have found that, as described above, sodium hyaluronate ophthalmic solution containing propylene glycol hardly causes dripping and has an effect of protecting corneal epithelial cells from drying.

That is, the present invention relates to an eye drop solution which is an aqueous eye drop solution containing hyaluronic acid or a salt thereof at a concentration of 0.03 to 0.5% (w/v) and propylene glycol at a concentration of 0.1 to 1.0% (w/v), wherein benzalkonium chloride at a concentration of 0.001 to 0.002% (w/v) is contained as a sole preservative, and an ionic isotonic agent at a concentration at which the osmotic pressure ratio of the eye drop solution becomes 0.9 to 1.1 (hereinafter, also simply referred to as "the present eye drop solution") is contained.

Another embodiment of the present invention is an eye drop containing a buffer agent and a pH adjuster in an amount to bring the pH to 6.0 to 7.0.

Another embodiment of the present invention is the present eye drops containing edetate at a concentration of 0.001 to 0.1% (w/v).

Another embodiment of the present invention is the eye drop wherein the concentration of hyaluronic acid or a salt thereof is 0.1 to 0.3% (w/v).

Another embodiment of the present invention is the eye drop wherein the concentration of propylene glycol is 0.25 to 0.75% (w/v).

In addition, the present invention provides the eye drops, wherein the concentration of benzalkonium chloride is 0.001-0.0015% (w/v).

Another embodiment of the present invention is an aqueous eye drop substantially comprising hyaluronic acid or a salt thereof at a concentration of 0.03 to 0.5% (w/v), propylene glycol at a concentration of 0.1 to 1.0% (w/v), benzalkonium chloride at a concentration of 0.001 to 0.002% (w/v), an ionic isotonicity agent at a concentration such that the osmotic pressure ratio of the eye drop is 0.9 to 1.1, a buffer agent and a pH adjuster in an amount such that the pH of the eye drop is 6.0 to 7.0, and edetate at a concentration of 0.001 to 0.1% (w/v).

Another embodiment of the present invention is an aqueous eye drop substantially comprising hyaluronic acid or a salt thereof at a concentration of 0.1 to 0.3% (w/v), propylene glycol at a concentration of 0.25 to 0.75% (w/v), benzalkonium chloride at a concentration of 0.001 to 0.0015% (w/v), an ionic isotonic agent at a concentration such that the osmotic pressure ratio of the eye drop is 0.9 to 1.1, a buffer agent and a pH adjusting agent in an amount such that the pH of the eye drop is 6.0 to 7.0, and edetate at a concentration of 0.001 to 0.1% (w/v).

The present invention also relates to a method for improving the storage efficiency of an aqueous eye drop containing sodium hyaluronate in a concentration of 0.1 or 0.3% (w/v) and benzalkonium chloride as a sole preservative, and for achieving a kinematic viscosity of the eye drop of 3.0 to 4.0 or 17 to 30mm²And/s and preventing dripping of the eye drops, the method comprising: a step (a) of mixing sodium hyaluronate in an amount to achieve a concentration of 0.1 or 0.3% (w/v) in the eye drop, benzalkonium chloride in an amount to achieve a concentration of 0.001-0.002% (w/v) in the eye drop, propylene glycol in an amount to achieve a concentration of 0.1-1.0% (w/v) in the eye drop, and an ionic isotonic agent in an amount to achieve an osmotic pressure ratio of 0.9-1.1 in the eye drop; a step (b) of adjusting the pH of the eye drop to 6.0 to 7.0; and a step (c) of filling the eye drops into an eye drop container (hereinafter, these are also collectively referred to simply as "the present method").

In the step (a) of the present method, edetate is preferably further mixed in an amount such that the concentration in the eye drops is 0.001 to 0.1% (w/v).

In the step (a) of the present method, it is preferable to mix propylene glycol so that the concentration in the eye drop is 0.25 to 0.75% (w/v).

In the step (a) of the present method, benzalkonium chloride is preferably mixed in an amount such that the concentration in the eye drop is 0.001 to 0.0015% (w/v).

The present eye drops have a kinematic viscosity and an osmotic pressure ratio acceptable as eye drops containing hyaluronic acid or a salt thereof (hereinafter also referred to simply as "hyaluronic acid eye drops") while exhibiting a sufficient storage effect by containing propylene glycol, even though the benzalkonium chloride concentration in the eye drops is 0.002% (w/v) or less. Further, as is clear from the results of the drip confirmation test and the corneal epithelial cell protective effect confirmation test described below, the present eye drops are less apt to cause dripping and also have an effect of protecting corneal epithelial cells from drying.

Drawings

Fig. 1 is a graph showing the effect of sodium hyaluronate eye drops containing propylene glycol on corneal epithelial cell damage caused by a drying load, with living cell activity on the vertical axis.

Detailed Description

The eye drop is an aqueous eye drop containing hyaluronic acid or a salt thereof at a concentration of 0.03-0.5% (w/v) and propylene glycol at a concentration of 0.1-1.0% (w/v), and is characterized by containing benzalkonium chloride at a concentration of 0.001-0.002% (w/v) as a sole preservative and an ionic isotonic agent at a concentration such that the osmotic pressure ratio of the eye drop is 0.9-1.1. In the present invention, the "aqueous eye drops" refer to eye drops containing water as a base.

The eye drops can be used not only for the treatment of corneal/conjunctival epithelial injuries accompanying endogenous diseases such as dry eye (xerophthalmia), Sjogren syndrome, Stevens-Johnson syndrome, etc., but also for the treatment of corneal/conjunctival epithelial injuries accompanying exogenous diseases caused by post-operation, drug-induced, trauma, contact lens wearing, etc. The present eye drops described above have a kinematic viscosity and an osmotic pressure ratio acceptable as hyaluronic acid eye drops, while they exhibit sufficient storage efficiency by containing propylene glycol, even though the benzalkonium chloride concentration in the eye drops is 0.002% (w/v) or less. Furthermore, the eye drop is not easy to generate dropping liquid, and has the effect of protecting corneal epithelial cells from drying.

The hyaluronic acid of the present invention is a compound represented by the following general formula (1).

[ in the formula, n represents a natural number. ]

The "hyaluronic acid" of the present invention is preferably hyaluronic acid having an average molecular weight of 50 to 390 ten thousand, and more preferably hyaluronic acid having an average molecular weight of 50 to 120 ten thousand.

The salt of hyaluronic acid is not particularly limited as long as it is a pharmaceutically acceptable salt, and may include salts with inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, sulfuric acid, and phosphoric acid; salts with organic acids such as acetic acid, fumaric acid, maleic acid, succinic acid, citric acid, tartaric acid, adipic acid, gluconic acid, glucoheptonic acid, glucuronic acid, terephthalic acid, methanesulfonic acid, lactic acid, hippuric acid, 1, 2-ethanedisulfonic acid, isethionic acid, lactobionic acid, oleic acid, methylenepamoic acid, polygalacturonic acid, stearic acid, tannic acid, trifluoromethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, lauryl sulfate, methyl sulfate, naphthalenesulfonic acid, and sulfosalicylic acid; quaternary ammonium salts with methyl bromide, methyl iodide, etc.; salts with halogen ions such as bromide ion, chloride ion, and iodide ion; salts with alkali metals such as lithium, sodium, and potassium; salts with alkaline earth metals such as calcium and magnesium; metal salts with iron, zinc, etc.; salts with ammonia; and salts with organic amines such as triethylenediamine, 2-aminoethanol, 2-iminobis (ethanol), 1-deoxy-1- (methylamino) -2-D-sorbitol, 2-amino-2- (hydroxymethyl) -1, 3-propanediol, procaine, and N, N-bis (phenylmethyl) -1, 2-ethylenediamine.

The "hyaluronic acid salt" of the present invention is preferably a sodium salt represented by the following general formula (2) (hereinafter also referred to as "sodium hyaluronate").

[ in the formula, m represents a natural number. ]

The "hyaluronic acid or a salt thereof" of the present invention may be in the form of a hydrate or a solvate.

In the case where hyaluronic acid exists as geometric isomers or optical isomers, the isomers or their salts are also included in the scope of the present invention. In addition, in the case where hyaluronic acid has proton tautomerism, the tautomer or a salt thereof is also included in the scope of the present invention.

In the case where hyaluronic acid or a salt, hydrate, or solvate thereof exists in a polymorphic form and a polymorphic group (polymorphic system), these polymorphic form and polymorphic group (polymorphic system) are also included in the scope of the present invention. The polymorphic group (polymorphic system) refers to various crystal forms at various stages when the crystal form changes due to conditions and states (including a preparation state) of the production, crystallization, storage, and the like of the crystals, and the whole process thereof.

"hyaluronic acid or a salt thereof" may be produced by a method conventionally used in the field of synthetic organic chemistry, or may be produced by a method described in Japanese patent application laid-open No. Hei 1-115902. In addition, "hyaluronic acid or a salt thereof" of the present invention may be hyaluronic acid or a salt thereof commercially available from Sigma or the like, and for example, "sodium hyaluronate" is commercially available from Sigma (catalog number: H5388).

The present eye drops may contain an active ingredient other than "hyaluronic acid or a salt thereof", but preferably contain "hyaluronic acid or a salt thereof" as the only active ingredient.

The concentration of the "hyaluronic acid or a salt thereof" in the eye drop is 0.03 to 0.5% (w/v), preferably 0.1 to 0.3% (w/v), and more preferably 0.1% (w/v) or 0.3% (w/v).

The "concentration of hyaluronic acid or a salt thereof" refers to both the concentrations of hyaluronic acid (free form) and hyaluronic acid salt. For example, "0.1% (w/v) hyaluronic acid or a salt thereof" refers to both the case where the concentration of "hyaluronic acid (free body)" in the eye drops is 0.1% (w/v) and the case where the concentration of "hyaluronic acid salt" is 0.1% (w/v).

The concentration of the "propylene glycol" in the eye drops is 0.1 to 1.0% (w/v), preferably 0.25 to 0.75% (w/v).

Benzalkonium chloride is a compound represented by the following general formula (3).

[ wherein R represents an alkyl group having 8 to 18 carbon atoms. ]

As the "benzalkonium chloride" of the present invention, it is preferable that R in the above general formula (3) represents "C₁₂H₂₅"a compound (hereinafter also referred to as" benzalkonium chloride (C12) ").

The concentration of benzalkonium chloride in the eye drops is 0.001-0.002% (w/v), preferably 0.001-0.0015% (w/v).

In the present invention, "containing benzalkonium chloride as the only preservative" means that the present eye drops contain benzalkonium chloride and do not contain any of benzethonium chloride, chlorhexidine gluconate, parabens (parabens), sorbic acid and its salts, chlorobutanol, boric acid, borax, and chlorite.

In the present invention, the phrase "containing an ionic isotonic agent in a concentration such that the osmotic pressure ratio of the eye drops is 0.9 to 1.1" means that the osmotic pressure ratio of the eye drops is adjusted to be in the range of "0.9 to 1.1" by adding an ionic isotonic agent in addition to propylene glycol or the like to the eye drops. The osmotic pressure ratio of the present eye drops can be easily measured using an automatic osmotic pressure analyzer.

The ionic isotonic agent used in the present invention is an ionic isotonic agent such as sodium chloride, potassium chloride, calcium chloride or magnesium chloride.

The eye drop preferably contains a buffer and a pH adjuster in amounts such that the pH of the eye drop is 6.0 to 7.0. In the present invention, the phrase "containing a buffering agent and a pH adjusting agent in an amount to bring the pH of the eye drop to 6.0 to 7.0" means that the pH of the eye drop is adjusted to a range of "6.0 to 7.0" by adding the buffering agent and the pH adjusting agent to the eye drop, and the amount (concentration) of the buffering agent and the pH adjusting agent to be added is not particularly limited as long as the pH of the eye drop can be adjusted to the range.

Specific examples of the "buffer" in the present invention include sodium phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium acetate, and aminocaproic acid, and particularly preferred is aminocaproic acid. Here, boric acid and borax are not included in the "buffer" of the present invention.

The "pH adjuster" in the present invention is not particularly limited as long as it can adjust the pH of the eye drop, and specific examples thereof include dilute hydrochloric acid, sodium hydroxide, and the like.

The eye drops have kinematic viscosity equal to that of the existing sodium hyaluronate eye drops. For example, when the concentration of hyaluronic acid or a salt thereof in the eye drops is 0.1% (w/v), the kinematic viscosity thereof is Hyalein^{(registered trademark)}Eye drops of 0.1% or Hyalein^{(registered trademark)}"3.0 to 4.0 mm" described in the summary of mini-eyedrops 0.1% pharmaceutical preparations²In the range of/s'. Further, when the concentration of hyaluronic acid or a salt thereof in the eye drops is 0.3% (w/v), the kinematic viscosity thereof is Hyalein^{(registered trademark)}Eye drops of 0.3% or Hyalein^{(registered trademark)}"17 to 30 mm" described in the summary of mini-eyedrops 0.3% pharmaceutical preparations²In the range of/s'.

The eye drop preferably contains 0.001-0.1% (w/v) edetate. The "edetate salt" in the present invention means a salt of edetic acid (ethylenediaminetetraacetic acid) such as monosodium edetate, disodium edetate, trisodium edetate, tetrasodium edetate, and the like, and a hydrate thereof. The "edetate" of the present invention is preferably disodium edetate dihydrate (hereinafter also referred to as "sodium edetate hydrate").

The present eye drops may contain pharmaceutically acceptable additives as needed, but the present eye drops preferably do not contain additives which are not specifically described and which affect the effects of the present eye drops, and more preferably do not contain additives which are not specifically described.

As a preferred embodiment of the above-mentioned eye drops, the present invention provides an aqueous eye drop substantially comprising hyaluronic acid or a salt thereof at a concentration of 0.03 to 0.5% (w/v), propylene glycol at a concentration of 0.1 to 1.0% (w/v), benzalkonium chloride at a concentration of 0.001 to 0.002% (w/v), an ionic isotonic agent at a concentration at which the osmotic pressure ratio of the eye drop is 0.9 to 1.1, a buffer agent and a pH adjusting agent in an amount at which the pH of the eye drop is 6.0 to 7.0, and edetate at a concentration of 0.001 to 0.1% (w/v). In the aqueous eye drops of the present embodiment, it is also preferable that the concentration of hyaluronic acid or a salt thereof is 0.1 to 0.3% (w/v), the concentration of propylene glycol is 0.25 to 0.75% (w/v), and the concentration of benzalkonium chloride is 0.001 to 0.0015% (w/v).

In the present method, the phrase "improving the storage efficiency" means that the sodium hyaluronate ophthalmic solution containing benzalkonium chloride at a concentration of 0.002% (w/v) or less is passed through a predetermined storage efficiency test (for example, the fifteenth modification of the storage efficiency test of the japanese pharmacopoeia) without increasing the amount of the preservative added.

In the present method, "prevention of dripping" means that the frequency of occurrence of a phenomenon (dripping) in which sodium hyaluronate eye drops drip from the tip of an eye drop container to the outside can be reduced in comparison with sodium hyaluronate eye drops not containing propylene glycol when an eye dropping operation is performed after the sodium hyaluronate eye drops are filled in the eye drop container.

In the step (a) of the method, ingredients other than sodium hyaluronate, benzalkonium chloride, propylene glycol and an ionic isotonizing agent may be mixed, and it is particularly preferable to mix edetate in the sodium hyaluronate eye drops in an amount of 0.001 to 0.1% (w/v).

The step (b) of the method is to add a buffering agent and a pH regulator into the sodium hyaluronate eye drops to adjust the pH of the sodium hyaluronate eye drops to be within the range of 6.0-7.0. The addition amount (concentration) of the buffer and the pH adjuster is not particularly limited as long as the pH of the sodium hyaluronate eye drops can be adjusted within the above range.

In the step (c) of the present method, the eye drop container to be filled with the sodium hyaluronate eye drops is not particularly limited as long as it is a container generally used as an eye drop container, and a polyethylene eye drop container is particularly preferable.

The method may also include steps other than steps (a), (b) and (c).

The results of the storage efficacy test, the kinematic viscosity measurement test, the drip confirmation test, and the corneal epithelial cell protective effect confirmation test, and preparation examples are shown below, and these examples are for better understanding of the present invention and do not limit the scope of the present invention.

Examples

[ preservation efficacy test ]

In order to confirm the influence of propylene glycol on the storage efficiency of hyaluronic acid eye drops, a storage efficiency test was performed.

(sample preparation)

< comparative formulations 1 and 2>

0.3g of sodium hyaluronate, 0.7g of sodium chloride, 0.15g of potassium chloride, 0.2g of aminocaproic acid, 0.01g of sodium edetate hydrate and 0.0025g of benzalkonium chloride (C12) were dissolved in water to a constant volume of 100mL, and diluted hydrochloric acid and/or sodium hydroxide were added to prepare a solution having a pH of 6.0 as comparative formulation 1 and a solution having a pH of 7.0 as comparative formulation 2.

< comparative formulations 3 and 4>

The preparation was carried out in the same manner as in comparative formulations 1 and 2, except that the amount of benzalkonium chloride (C12) added was 0.002 g.

< formulation 1>

0.3g of sodium hyaluronate, 0.7g of sodium chloride, 0.25g of propylene glycol, 0.2g of aminocaproic acid, 0.01g of sodium edetate hydrate and 0.001g of benzalkonium chloride (C12) were dissolved in water to a constant volume of 100mL, and diluted hydrochloric acid and/or sodium hydroxide was added thereto to adjust the pH to 6.0.

< formulation 2>

The preparation was carried out in the same manner as in the above formulation 1 except that the amount of sodium hyaluronate was 0.1g and the amount of benzalkonium chloride (C12) was 0.0012 g.

(test method)

The preservation efficacy test was performed based on the fifteenth revised preservation efficacy test method of the japanese pharmacopoeia (hereinafter also simply referred to as "japanese pharmacopoeia"). In this test, escherichia coli (e.coli), pseudomonas aeruginosa (p.aeruginosa), staphylococcus aureus (s.aureus), candida albicans (c.albicans), and aspergillus niger (a.niger) were used as test bacteria.

(test results)

The test results are shown in Table 1. In table 1, "n.d." indicates an undetected bacterium.

[ Table 1]

(discussion)

As shown in table 1, it was found that the sodium hyaluronate ophthalmic solution satisfied the criterion of "japanese pharmacopoeia reference information storage efficacy test type IA" in the range of acceptable pH (pH6.0 and 7.0) in the presence of 0.0025% (w/v) benzalkonium chloride, but did not satisfy the criterion at pH6.0 in the presence of 0.002% (w/v) benzalkonium chloride, and did not have sufficient storage efficacy. Then, the preservation efficacy at ph6.0 at which the preservation efficacy became weak was evaluated in the presence of 0.25% (w/v) propylene glycol, and it was found that 0.1% (w/v) sodium hyaluronate ophthalmic solution satisfied the criterion of "japanese pharmacopoeia reference information preservation efficacy test type IA" at a benzalkonium chloride concentration of 0.0012% (w/v), and 0.3% (w/v) sodium hyaluronate ophthalmic solution also satisfied the criterion at a benzalkonium chloride concentration of 0.001% (w/v), and each had sufficient preservation efficacy. As described above, the hyaluronic acid ophthalmic solution can exhibit a sufficient storage efficiency even when the benzalkonium chloride concentration is 0.002% (w/v) or less by adding propylene glycol.

[ kinematic viscosity measurement test ]

In order to confirm the kinematic viscosity of the hyaluronic acid eye drops containing propylene glycol when the osmotic pressure ratio of the same is adjusted to 0.9 to 1.1, a kinematic viscosity measurement test was performed.

(sample preparation)

< comparative formulation >

0.3g of sodium hyaluronate, 0.2g of aminocaproic acid, 0.8g of sodium chloride, 0.0015g of benzalkonium chloride (C12), and 0.01g of sodium edetate hydrate were dissolved in water to a constant volume of 100mL, and diluted hydrochloric acid and/or sodium hydroxide was added to adjust the pH to 6.5 and the osmotic pressure ratio to 1.0, which was used as a comparative formulation.

< formulations 1 to 5>

The preparation was carried out in the same manner as in the comparative formulation except that the osmotic pressure ratio was adjusted to 1.0 by appropriately changing the amounts of sodium chloride and propylene glycol added (see table 2).

(test method)

On page 4 of the Hyalein general description, the allowable ranges of osmotic pressure ratio and kinematic viscosity of 0.3% (w/v) sodium hyaluronate eye drops are "0.9 to 1.1" and "17 to 30 mm", respectively²And/s ". Then, the osmotic pressure ratio of the sodium hyaluronate eye drops is adjusted to "0.9 to 1.1" by using propylene glycol and an ionic isotonic agent, and then the kinematic viscosity of the eye drops is measured. The kinematic viscosity measurement test was carried out by measuring the kinematic viscosity at a measurement temperature of 30 ℃ according to the "first capillary viscometer method of viscometry according to the conventional test method of the japanese pharmacopoeia". The osmotic pressure ratio was measured by an automatic osmotic pressure analyzer (manufactured by ARKRAY) in accordance with "osmotic pressure measurement method based on the general test method in Japanese pharmacopoeia".

(results)

The results are shown in Table 2.

[ Table 2]

Compounding ingredients

Comparative formulations

Formulation 1

Formulation 2

Formulation 3

Formulation 4

Formulation 5

Hyaluronic acid sodium salt

0.3％

0.3％

0.3％

0.3％

0.3％

0.3％

ε Aminocaproic acid

0.2％

0.2％

0.2％

0.2％

0.2％

0.2％

Sodium chloride

0.8％

0.7％

0.6％

0.5％

0.4％

0.3％

Propylene glycol

-

0.25％

0.5％

0.75％

1％

1.25％

Benzalkonium chloride (C12)

0.0015％

0.0015％

0.0015％

0.0015％

0.0015％

0.0015％

Edetate sodium hydrate

0.01％

0.01％

0.01％

0.01％

0.01％

0.01％9 -->

Dilute hydrochloric acid/sodium hydroxide

Proper amount of

Proper amount of

Proper amount of

Proper amount of

Proper amount of

Proper amount of

Purified water

Proper amount of

Proper amount of

Proper amount of

Proper amount of

Proper amount of

Proper amount of

pH

6.5

6.5

6.5

6.5

6.5

6.5

Osmotic pressure ratio

1.0

1.0

1.0

1.0

1.0

1.0

Kinematic viscosity (mm)2/s)

25

26

27

28

30

34

(discussion)

As is clear from Table 2, when the propylene glycol content is 1% (w/v) or less, the kinematic viscosity of the sodium hyaluronate eye drops of 0.3% (w/v) is within the allowable range (17 to 30 mm)²In/s) and, on the other hand, at concentrations above that, outside the permitted range of kinematic viscosity. Furthermore, from the above results, it is presumed that, when the blending concentration of propylene glycol is 1% (w/v) or less, the kinematic viscosity of 0.1% (w/v) sodium hyaluronate eye drops is within the allowable range (3.0 to 4.0 mm)²In s). As described above, when propylene glycol is added to hyaluronic acid eye drops, the addition concentration thereof needs to be 1.0% (w/v) or less.

[ test for confirming dropping liquid ]

In order to confirm the influence of propylene glycol on the dropping of hyaluronic acid eye drops, a dropping confirmation test was performed.

(reagent preparation)

Formulations without propylene glycol

Use of commercially available "Hyalein^{(registered trademark)}Eye drops 0.1% ".

Formulations containing propylene glycol

Sodium hyaluronate (0.1g), buffer (-aminocaproic acid), sodium chloride, propylene glycol, benzalkonium chloride (C12) and sodium edetate hydrate were dissolved in water to a constant volume of 100mL (osmotic pressure ratio: 0.9-1.1), and dilute hydrochloric acid and/or sodium hydroxide were added to bring the pH to 6.5.

(test method)

The polyethylene eye drop container was filled with 5mL of a formulation containing no propylene glycol or a formulation containing propylene glycol, and the same operation as that in eye drop was performed 6 times by 5 healthy persons. For each operation, whether dripping occurred (phenomenon in which eye drops drip from the tip of the eye drop container to the outside) was evaluated.

(results)

The results are shown in Table 3.

[ Table 3]

(discussion)

As can be seen from table 3, dripping sometimes occurred in the formulation containing no propylene glycol, but dripping was not observed at all in the formulation containing propylene glycol. In conclusion, the eye drops have improved dropping properties compared to the conventional hyaluronic acid eye drops not containing propylene glycol.

[ confirmation test for corneal epithelial cell protective Effect ]

In order to examine the effect of the hyaluronic acid eye drops containing propylene glycol on damage of corneal epithelial cells caused by a drying load, a test for confirming the protective effect of corneal epithelial cells was performed.

(test method)

SV40 immortalized human corneal epithelial cells (HCE-T: institute of physico-chemical engineering, center of biological resources, cell number: RCB2280) were inoculated into a 96-well plate (1 × 10)⁴Individual cells/well) were cultured with SHEM medium for 1 day. The following day, the medium was changed to D-MEM/F12 medium containing 0.25% (w/v) propylene glycol, 0.03% (w/v) sodium hyaluronate, or 0.25% (w/v) propylene glycol and 0.03% (w/v) sodium hyaluronate, and then the corneal epithelial cells were cultured for 1 hour (hereinafter, also referred to as "propylene glycol single group", "hyaluronic acid single group", or "propylene glycol/hyaluronic acid combined group", respectively). After the exchange into D-MEM/F12 medium containing no test substance, the corneal epithelial cells were cultured for 1 hour, and this group was referred to as "stromal group". After the culture, the medium of each group was changed to D-MEM/F12 medium containing no test substance, and then a drying load was applied for 7.5 minutes. After the load, the activity of viable cells (absorbance at 490 nm) was measured using a cell proliferation assay kit (CellProlifications assay kit) (catalog number: G3580, manufactured by Promega corporation)

(results)

The test results are shown in FIG. 1.

(discussion)

As is clear from fig. 1, the decrease in the activity of the viable cells of the corneal epithelial cells (see the stromal group) due to the drying load was not sufficiently suppressed by the treatment with propylene glycol or hyaluronic acid alone (see the propylene glycol alone group and the hyaluronic acid alone group). On the other hand, it was surprisingly confirmed that the decrease in the activity of the living cells was significantly suppressed in the propylene glycol/sodium hyaluronate combined group. From the above results, it is considered that the hyaluronic acid eye drops containing propylene glycol have an effect of protecting corneal epithelial cells from dryness.

[ preparation examples ]

The pharmaceutical agent of the present invention will be described more specifically with reference to formulation examples, but the present invention is not limited to these formulation examples.

(formulation example 1)

Sodium hyaluronate and other components are added to sterile purified water and sufficiently mixed to prepare 0.1% (w/v) sodium hyaluronate eye drops having a pH of 6.0 to 7.0 and an osmotic pressure ratio of 0.9 to 1.1.

(formulation example 2)

Sodium hyaluronate and other components are added to sterile purified water and sufficiently mixed to prepare 0.3% (w/v) sodium hyaluronate eye drops having a pH of 6.0 to 7.0 and an osmotic pressure ratio of 0.9 to 1.1.

Industrial applicability

The hyaluronic acid ophthalmic solution of the present invention has an osmotic pressure ratio and a kinetic viscosity acceptable as a hyaluronic acid ophthalmic solution, while it exhibits a sufficient storage effect by containing propylene glycol, even though the benzalkonium chloride concentration in the ophthalmic solution is 0.002% (w/v) or less. Further, the hyaluronic acid eye drops of the present invention are not easily dropped and have an effect of protecting corneal epithelial cells from dryness, and therefore, are expected to be eye drops capable of more effectively treating dry eye.

Claims (10)

1. An eye drop comprising hyaluronic acid or a salt thereof at a concentration of 0.03 to 0.5% (w/v) and propylene glycol at a concentration of 0.1 to 1.0% (w/v), wherein the eye drop comprises benzalkonium chloride at a concentration of 0.001 to 0.0015% (w/v) as the only preservative and an ionic isotonic agent at a concentration such that the osmotic pressure ratio of the eye drop is 0.9 to 1.1.

2. An ophthalmic solution according to claim 1, which contains a buffering agent and a pH adjusting agent in amounts such that the pH of the ophthalmic solution is 6.0 to 7.0.

3. An ophthalmic solution according to claim 1 or 2, wherein the edetate is contained at a concentration of 0.001-0.1% (w/v).

4. An ophthalmic solution according to claim 1, wherein the concentration of hyaluronic acid or a salt thereof is 0.1 to 0.3% (w/v).

5. An ophthalmic solution according to claim 1, wherein the concentration of propylene glycol is 0.25 to 0.75% (w/v).

6. An aqueous eye drop substantially comprising hyaluronic acid or a salt thereof at a concentration of 0.03 to 0.5% (w/v), propylene glycol at a concentration of 0.1 to 1.0% (w/v), benzalkonium chloride at a concentration of 0.001 to 0.0015% (w/v), an ionic isotonic agent at a concentration such that the osmotic pressure ratio of the eye drop is 0.9 to 1.1, a buffer agent and a pH adjusting agent in an amount such that the pH of the eye drop is 6.0 to 7.0, and edetate at a concentration of 0.001 to 0.1% (w/v).

7. An aqueous eye drop containing sodium hyaluronate at a concentration of 0.1% (w/v) and benzalkonium chloride as a sole antiseptic has improved storage efficiency and has a kinematic viscosity of 3.0-4.0 mm²A method for preventing dripping of the eye drops, the method comprising the steps of:

step (a): mixing sodium hyaluronate in an amount to achieve a concentration of 0.1% (w/v) in the eye drops, benzalkonium chloride in an amount to achieve a concentration of 0.001-0.0015% (w/v) in the eye drops, propylene glycol in an amount to achieve a concentration of 0.1-1.0% (w/v) in the eye drops, and an ionic isotonic agent in an amount to achieve an osmotic pressure ratio of 0.9-1.1 in the eye drops;

step (b): adjusting the pH value of the eye drops to 6.0-7.0; and

step (c): the eye drops are filled in an eye drop container.

8. An aqueous eye drop containing sodium hyaluronate at a concentration of 0.3% (w/v) and benzalkonium chloride as a sole antiseptic has improved storage efficiency and kinematic viscosity of 17-30 mm²A method for preventing dripping of the eye drops, the method comprising the steps of:

step (a): mixing sodium hyaluronate in an amount to achieve a concentration of 0.3% (w/v) in the eye drops, benzalkonium chloride in an amount to achieve a concentration of 0.001-0.0015% (w/v) in the eye drops, propylene glycol in an amount to achieve a concentration of 0.1-1.0% (w/v) in the eye drops, and an ionic isotonic agent in an amount to achieve an osmotic pressure ratio of 0.9-1.1 in the eye drops;

step (b): adjusting the pH value of the eye drops to 6.0-7.0; and

step (c): the eye drops are filled in an eye drop container.

9. The method according to claim 7 or 8, wherein in step (a), edetate is further mixed in an amount such that the concentration in the eye drops is 0.001-0.1% (w/v).

10. The method according to claim 7 or 8, wherein in step (a), the propylene glycol is mixed in an amount such that the concentration in the eye drop is 0.25 to 0.75% (w/v).