JP2001240536A - Perfusate preparation for ophthalmic operation - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an intraoculo perfusate preparation which is excellent in protection of ocular tissue including endothelial cell and sustained effect of retinal function during and after operation in an ophthalmic operation and high safety for organism. SOLUTION: The perfusate preparation for ophthalmic operation results in high safety and excellent protection of ocular tissue including corneal endothelial cell and sustained effect of retinal function by combining D-3- hydroxybutyric acid and its salts and bicarbonate as an energy source. A stable preparation can be achieved by adding glucose, buffer and stabilizer or the like.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a perfusion solution for eye surgery such as cataract surgery, intraocular lens transplantation surgery, glaucoma surgery, vitreous surgery and full thickness corneal transplantation surgery. More specifically, in order to perform the operation safely and effectively,
The present invention relates to a perfusion solution for ophthalmic surgery suitable for protecting intraocular tissues, aspirating and removing surgical residues in the eye, and preventing corneal epithelium and conjunctiva from drying.

[0002]

2. Description of the Related Art In recent years, the development of ophthalmic surgical modes such as cataract surgery, intraocular lens transplantation surgery, and glaucoma surgery has made remarkable progress. In order to safely and effectively perform the above-mentioned operation, a perfusion solution used as an auxiliary for surgery plays an important role. For example, at the time of cataract surgery, the aqueous humor is drained from the anterior chamber by incising the periphery of the cornea with a scalpel at the start of the operation. Injection of a viscoelastic material is essential. Furthermore, if a perfusate is present in the eye during the crushing and excision of the opaque lens, the crushed material can be smoothly removed by suction. In addition, a perfusate is used to prevent drying of the keratoconjunctival surface during surgery.

[0003] In order to achieve the purpose of using such perfusate, precautions in formulation are: 1) Osmotic pressure and pH
Is physiologically harmonious with intraocular tissues and endothelial cells; 2) adding the minimum required compounds of aqueous humor components such as inorganic salts, energy sources and cell activators; 3) biologically safe And 4) the preparation can be stored at room temperature for a long period of time.

[0004] At present, perfusion solutions for eye surgery marketed in Japan include products containing oxyglutathione as a cell activator and products having a buffer system based on bicarbonate ions. However, these are two-part preparations that must be mixed at the time of use, and because they do not contain components effective for protecting the corneal endothelium,
It contains not only problems of formulation stability and ease of use, but also problems to be solved in terms of efficacy.

Further, a perfusion solution for ophthalmic surgery containing 3-hydroxybutyric acid as a main energy source is disclosed in US Pat. No. 5,116,868 and US Pat. No. 5,298,4.
No. 87 discloses it. However, the preparation described in this patent publication does not contain bicarbonate ions necessary to maintain the function of the corneal endothelium. The reason is that when bicarbonate ion is present in the preparation, the pH of the preparation fluctuates and becomes unstable due to the partial pressure of CO ₂ , and 3-hydroxybutyric acid automatically converts CO ₂ generated by metabolism into bicarbonate ion And therefore it is not necessary to premix the bicarbonate ions in the formulation.

[0006] D-3-hydroxybutyric acid or a salt thereof used as an active ingredient in the present invention is known as a biological component, is produced by oxidation of fatty acids in the liver, and is used as an energy source in peripheral tissues. (Lenininger Shinsei Chemistry, 2nd edition, p62
5, 1993). It is also known that this substance is extremely useful as an energy-producing substrate for corneal tissue rather than glucose (TRANSPLANTA).
TION, 57, pp. 1778-1785, 1994). The application of D-3-hydroxybutyric acid as a pharmaceutical has been reported as an infusion composition component for nutritional supplementation in patients with a state of increased biocatabolism or invasion of the body (see JP-A-2-191212). ).

[0007]

SUMMARY OF THE INVENTION An object of the present invention is to provide an intraocular perfusion solution which is excellent in protection of intraocular and postoperative ocular tissues and endothelial cells during eye surgery and has high biosafety. . Another object of the present invention is to add inorganic ions, an energy source, a tonicity agent, a buffering agent, a bicarbonate ion and a stabilizing agent, etc., in addition to the above-mentioned excellent properties, and to further improve the pharmaceutically stability. To provide a novel intraocular perfusion solution formulation.

[0008]

Means for Solving the Problems The present inventors have conducted intensive studies to solve the above-mentioned problems, and as a result, by combining D-3-hydroxybutyric acid or its salts used as an energy source with bicarbonate. An inorganic salt, an isotonic agent, and glucose, which are highly safe as a perfusion solution preparation for ophthalmic surgery and exhibit excellent functions for protecting ocular tissues and endothelial cells, and are known as human aqueous humor components. Was added, and furthermore, a buffer and a stabilizer were added to clarify that a stable preparation could be provided, and the present invention was completed.

That is, according to the present invention, the above objects and advantages of the present invention are achieved by the following perfusion solution preparation for ophthalmic surgery. (1) (A) at least one compound selected from the group consisting of D-3-hydroxybutyric acid and salts thereof in a concentration of 0.1 mM to 500 mM in an aqueous solution, and (B) bicarbonate in a concentration of an aqueous solution. 0.1 mM to 100 mM and (C) 0.1 mM as glucose in the aqueous solution.
A perfusion solution preparation for ophthalmic surgery, comprising an aqueous solution containing ５０50 mM. (2) (A) at least one compound selected from the group consisting of D-3-hydroxybutyric acid and salts thereof in a concentration of 0.1 mM to 500 mM in an aqueous solution, and (B) bicarbonate in a concentration of an aqueous solution. 0.1 mM to 100 mM and (C) 0.1 mM as glucose in the aqueous solution.
A perfusion solution for eye surgery, comprising an aqueous solution containing ５０50 mM, having a pH of 6.8 to 8.2, and an osmotic pressure of 270 to 350 mOsm. (3) (A) at least one compound selected from the group consisting of D-3-hydroxybutyric acid and salts thereof in a concentration of 0.1 mM to 500 mM in an aqueous solution, and (B) bicarbonate in a concentration of an aqueous solution. 0.1 mM to 100 mM,
(C) 0.1 mM or more of glucose as a concentration in an aqueous solution.
The concentration of 50 mM and (D) citric acid and / or an alkali metal salt thereof is 0.01 mM to 7.
A perfusion solution for ophthalmic surgery, comprising an aqueous solution containing 5 mM, having a pH of 6.8 to 8.2 and an osmotic pressure of 270 to 350 mOsm. (4) (A) 0.1 mM to 500 mM of at least one compound selected from the group consisting of D-3-hydroxybutyric acid and salts thereof in an aqueous solution, and (B) bicarbonate in an aqueous solution. 0.1 mM to 100 mM and (E) a phosphate buffer in a concentration of 0.1
A perfusion solution for ophthalmic surgery, comprising an aqueous solution containing from 50 mM to 50 mM. (5) (A) at least one compound selected from the group consisting of D-3-hydroxybutyric acid and salts thereof in a concentration of 0.1 mM to 500 mM in an aqueous solution, and (B) bicarbonate in a concentration of an aqueous solution. 0.1 mM to 100 mM,
(C) 0.1 mM or more of glucose as a concentration in an aqueous solution.
50 mM, (D) citric acid and / or an alkali metal salt thereof in a concentration of 0.01 mM to 7.5 m
A perfusion solution for ophthalmic surgery, comprising an aqueous solution containing M and (E) a phosphate buffer in a concentration of 0.1 mM to 50 mM in the aqueous solution.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION With respect to the configuration at the C3 position in the chemical structural formula of 3-hydroxybutyric acid, there are a group of D-form, D, L-racemate and L-form. Among them, in the present invention, the D-form is used for maximizing the effectiveness of the perfusion solution preparation for eye surgery. D-3-hydroxybutyric acid can be contained in the preparation of the present invention as DL-form in addition to D-form. Since D-3-hydroxybutyric acid and its salt are easily synthesized by selective asymmetric catalytic reduction of ethyl acetoacetate and subsequent ester hydrolysis, the compound can be obtained at relatively low cost. In the present invention, D-3-hydroxybutyric acid may be contained as salts. The salts are preferably selected from the group consisting of sodium salts, potassium salts, L-lysine salts, L-histidine salts and L-arginine salts. The concentration of D-3-hydroxybutyric acid and its salt in the aqueous solution preparation of the present invention needs to be in the range of 0.1 to 500 mM.

In the preparation of the present invention, it is essential to contain a bicarbonate. As the bicarbonate, sodium bicarbonate and potassium bicarbonate are preferable, and the concentration thereof is in the range of 0.1 to 100 mM. is necessary. In the preparation without bicarbonate ion, the effect of suppressing corneal swelling, which is one of the important indicators for maintaining corneal function, is extremely small.

In the perfusion solution preparation of the present invention, in addition to the above components, inorganic salts present in human aqueous humor and vitreous components, glucose as another energy source, osmotic pressure and pH are coordinated with intraocular tissues and endothelial cells. It is desirable to appropriately include a tonicity agent and a buffering agent for the preparation, and a stabilizer for the preparation.

According to the present invention, the main energy source of the perfusate formulation is D-3-hydroxybutyric acid, but glucose may be supplemented as a secondary energy source to increase the effect of the present invention. . The glucose concentration is
A concentration range of 0.1-50 mM is preferred. As the stabilizer, for example, citric acid and its sodium salt and potassium salt are preferable, and the concentration thereof is 0.01 to 7.5 mM.
Is preferable. More preferably, 0.1 to 5.0.
It is in the range of mM.

The inorganic salt and the tonicity agent used in the present invention include, for example, inorganic salts such as alkali or alkaline earth metal salts composed of sodium chloride, potassium chloride, calcium chloride, magnesium chloride and magnesium sulfate; And mannitol, sorbitol,
Isotonic agents such as carbohydrates, such as xylitol and dextran, are preferably used. These can be used alone or in combination of two or more. The concentration of each of the inorganic salt and the tonicity agent is preferably in the range of 0.1 to 1,000 mM. The osmotic pressure of the preparation is 270-350 m
It is desirable to keep in the range of Osm.

Examples of the buffer include phosphate buffers such as disodium monohydrogen phosphate, sodium dihydrogen phosphate, dipotassium monohydrogen phosphate and potassium dihydrogen phosphate, and boric acid such as boric acid and sodium borate System buffers. Among them, a phosphate buffer present as a human aqueous humor component is preferable. The concentration of the buffer is
A range from 0.1 to 50 mM is preferred. The pH range of the aqueous solution preparation of the present invention is pH 6.8 to maintain the function of the ocular tissues and cells without causing any trouble.
8.2 is preferred. Further, the pH is preferably in the range of 7.2 to 8.0, and as a result of a functional test and a safety test using rabbit eyes, these pH ranges are acceptable as a perfusion solution for ophthalmic surgery.

Such a main component and an additive are dissolved in distilled water in a predetermined amount in order, and the pH is adjusted with dilute hydrochloric acid or a dilute alkali solution. Then, a transparent glass bottle or a plastic bottle is used as a one-pack type. And both can be stored under a sealed stopper. A sample in which the formulation thus stored was stored at 40 ° C. and 75% relative humidity for 6 months was compared with that at the start of the test,
There was no change in appearance or osmotic pressure. On the other hand, as for pH, the amount of sodium citrate is preferably 0.1 to 5.
When the concentration is set to 0 mM, more preferably 0.15 to 3.0 mM, the pH of the preparation at the start of the test becomes 7.3 to 3.0 mM.
At 7.4, the pH value of the aqueous solution is relatively small since the pH value is 7.4 to 7.8 even at the end of the preservation, and a relatively stable perfusate preparation can be obtained even in the presence of bicarbonate ions. .

[0017]

EXAMPLES Hereinafter, the present invention will be described specifically with reference to Examples and Comparative Examples, but the present invention is not limited to these.

Example 1 Solution No. No. 1 (the present invention), No. 2 (Comparative Example) and No. 2 3 (Comparative Example) is to dissolve predetermined amounts of the following components in distilled water in order from the top, and finally dissolve sodium D-3-hydroxybutyrate (hereinafter abbreviated as D-3-HBA) to obtain a total amount. A 1 L aqueous solution was prepared, the pH was adjusted with dilute hydrochloric acid, and then sterile filtered to obtain a test perfusion solution. The components of commercial product B (comparative solution 1) and commercial product M (comparative solution 2) used as comparative examples and their concentrations are also shown below.

Solution No. 1 (Product of the present invention) NaCl 100.3 mM KCl 5.1 mM MgCl ₂ 1.0 mM CaCl ₂ 1.1 mM Na ₂ HPO ₄ 3.0 mM Sodium citrate 0.34 mM NaHCO ₃ 25.0 mM D-3-HBA 20.0 mM Glucose 5.0 mM solution no. 2 (Comparative Example) NaCl 100.0 mM MgCl ₂ 1.0 mM CaCl ₂ 2.0 mM K ₂ HPO ₄ 5.0 mM Na ₂ HPO ₄ 5.0 mM sodium citrate 10.0 mM sodium acetate 20.0 mM D, L-3- HBA 10.0 mM Glucose 5.5 mM Solution No. 3 (Comparative Example) NaCl 100.3 mM KCl 5.1 mM MgCl ₂ 1.0 mM CaCl ₂ 1.1 mM Na ₂ HPO ₄ 3.0 mM Sodium citrate 0.34 mM NaHCO ₃ 25.0 mM Glucose 5.0 mM Comparative solution 1 (Comparative) Example) NaCl 122.2 mM KCl 5.08 mM MgCl ₂ 0.98 mM CaCl ₂ 1.05 mM Na ₂ HPO ₄ 3.0 mM NaHCO ₃ 25.0 mM Oxyglutathione 0.30 mM Glucose 5.11 mM Comparative solution 2 (Comparative example) NaCl 112 .9mM KCl 4.8mM CaCl ₂ 1.2mM sodium citrate 3.4 mM NaHCO ₃ 25.0 mm sodium acetate 4.4mM glucose 8.3mM standard perfusion solution (Comparative example) NaCl 120.0mM KCl 3.1mM CaCl ₂ · _{H 2 O 1.15mM MgSO 4 · H} 2 O 1.2mM KH 2 PO 4 0.5mM NaHCO 3 23.0mM glucose 10.0mM

Example 2 The solution No. 1 containing D-3-HBA of Example 1 1 (inventive product) was subjected to a preparation stability test. 500 mL of the preparation
Was filled into a transparent glass container having a capacity of 600 mL, and the container was sealed to prepare 5 test liquids. These are at 40 ° C. ± 0.2.
The temperature was adjusted to 5 ° C. and stored in a thermo-hygrostat adjusted to 75 ± 5% RH for 6 months. As a result, the appearance of the solution, the insoluble foreign matter test, and the osmotic pressure did not change. Solution pH
The pH at the start of storage was 7.3 to 7.4, but after 6 months of storage, the pH was 7.4 to 7.8, and the fluctuation was relatively small. It was found that it could be saved.

Example 3 The solution No. 1 described in Example 1 was used. No. 1 (the present invention), 2
Using four types of test perfusion solutions, (Comparative Example), Comparative Solution 1 (Comparative Example), and Comparative Solution 2 (Comparative Example), the effects of the protective action on the cornea were compared in an in vitro experiment. In a state where the sclera adhered to the periphery of the cornea from an adult rabbit, a scleral corneal piece was excised (total of 20 specimens for each of the four kinds of test liquids, 5 specimens).
Corneal thickness in a test perfusion solution at 36 ° C. for 5 hours using an ultrasonic keratometer (DGH-500 PACHE).
TTE, DGH TECHNOLOGY). FIG. 1 shows the change in corneal thickness of each test solution after culturing for 5 hours (= corneal thickness before culturing−corneal thickness after culturing for 5 hours).

Solution No. D-3 containing D-3-HBA and bicarbonate was mixed. Solution No. 1 (product of the present invention) is a solution No. 2 (Comparative Example) shows a remarkable effect on corneal protection compared to Comparative Solution 1 (Comparative Example) and Comparative Solution 2 (Comparative Example). Further suppression effect was shown.

Example 4 The test perfusate No. 1 described in Example 1 was used. 1 (product of the present invention), N
o. 2 (Comparative Example), Comparative Solution 1 (Comparative Example) and Comparative Solution 2
Dutchrabb using the same four types (Comparative Example)
The effect of perfusion in the anterior chamber on the corneal thickness in an in vivo experiment was examined by it (male and female, body weight: 1.9 to 2.9 Kg). After general anesthesia of each individual rabbit with xylazine hydrochloride and ketamine hydrochloride by intramuscular injection, a 3.2 mm wide incision was made in the limbus of the left and right eyes of each individual using an ophthalmic scalpel, and the tip of the needle was rounded. The cut 18G injection needle was inserted into the anterior chamber through the incision, and the four kinds of test perfusion fluids were each perfused at a flow rate of 10 mL / min for 120 minutes. The amount of change in the corneal thickness of each test solution was 1 before the perfusion and every 30 minutes after the start of the perfusion.
Up to 20 minutes, the measurement was performed using the above-mentioned ultrasonic keratometer under ophthalmic anesthesia with 0.4% oxyprocaine hydrochloride. FIG. 2 shows the amount of change with time of each corneal thickness.

As a result, in vitro of Example 3
Similarly to the experimental results, FIG. 2 shows that the solution containing D-3-HBA and bicarbonate contained no. No. 1 (the product of the present invention) is the solution No. 1 containing no bicarbonate. Compared with the perfusion solutions of Comparative Solution 2 (Comparative Example), Comparative Solution 1 (Comparative Example), and Comparative Solution 2 (Comparative Example), the perfusion solution was significantly smaller in the change in corneal thickness, and showed an excellent corneal protection effect. .

Example 5 The test perfusate No. 1 described in Example 1 was used. 1 (product of the present invention), N
o. In order to investigate the effect on the resected rabbit retina using three types of Comparative Example 3 and Comparative Solution 1 (Comparative Example), an electrophysiological test using an electroretinogram (hereinafter abbreviated as ERG) was performed as follows. Performed according to law. The retina extracted from the posterior part of the rabbit eyeball is set in a chamber that reacts with the test solution,
It was immersed in a standard perfusate at 35 ° C. aerated with a gas of 95% O ₂ : 5% CO ₂ until the ERG waveform was stable. When the ERG waveform was stabilized, the perfusion solution was replaced with the test perfusion solution kept at the same temperature and cultured for 60 minutes, and then replaced with the standard perfusion solution again and cultured for 60 minutes, and the b-wave amplitude of ERG was observed at 10-minute intervals. The fluctuation rate of the b-wave amplitude of ERG is shown in FIG. 3 with respect to the b-wave amplitude immediately before the replacement of the test liquid, and the change over time of each test liquid obtained by converting the amplitude fluctuation of each measurement time into percentage is shown in FIG.

As a result, the solution N containing D-3-HBA
o. No. 1 is a solution No. 1 containing no compound. As compared with Comparative Sample No. 3 and the commercially available Comparative Solution 1, the decrease in amplitude was the smallest, indicating that it was excellent in maintaining the retinal function. Further, even after re-substitution with the standard perfusion solution, the solution No. 1 (the product of the present invention) recovered the amplitude% to nearly 100% at all measurement times as compared with the other test solutions (comparative examples), indicating that the degree of recovery to normal retinal function was high. .
Therefore, the solution containing the D-3-HBA of the present invention, ie, the solution No. It can be said that No. 1 has an excellent function as a perfusate for vitreous surgery and has relatively little influence on the retinal function. ERG is a change in retinal function, that is, the degree of decrease in the number of retinal cells, in other words, the range of symptoms, the degree of qualitative and quantitative abnormalities of visual and neurotransmitters, abnormalities of retinal cell membrane potential, It is one of the effective methods for obtaining comprehensive information on the retina such as the degree of degeneration (Ophthalmology, Vol. 1, pp. 69-70, published by Nakayama Shoten,
1993).

[0027]

According to the present invention, D- is used as an energy source.
By combining 3-hydroxybutyric acid and its salts with bicarbonate, an aqueous solution that is highly safe as a perfusion solution for ophthalmic surgery and is excellent in protection of ocular tissues including corneal endothelial cells and maintenance of retinal function A stable perfusion solution for ophthalmic surgery can be prepared by adding the above-mentioned glucose, buffer and stabilizer to the formulation.

[Brief description of the drawings]

1 shows the protective action (in vitro experiment) on the cornea of the formulation solution of the present invention (solution No. 1) and the comparative formulation solution (solution No. 2, comparative solution 1, comparative solution 2).

FIG. 2 shows the amount of change over time (in vivo experiment) with respect to the corneal thickness of each formulation solution.

FIG. 1 (product of the present invention), solution no. Three
The ERG of the Comparative Example and Comparative Solution 1 (Comparative Example) over time is shown.

Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat II (Reference) A61K 47/12 A61K 47/12 A61P 27/02 A61P 27/02 (72) Inventor Shigeru Nakamura Kamimidai, Toyooka, Hyogo Prefecture 156-5 In Offtex Research Institute, Inc. (72) Inventor Toshihisa Hata 156-5, Kamidaidai, Toyooka City, Hyogo Prefecture (72) Inventor Fumio Saito 156, Kamimidai, Toyooka City, Hyogo Prefecture No. 5 Inside the Offtex Research Institute (72) Inventor Takeshi Onuma 2-17-8 Nihonbashi Hamacho, Chuo-ku, Tokyo Inside Offtex Corporation

Claims (5)

[Claims] (A) a concentration of at least one compound selected from the group consisting of D-3-hydroxybutyric acid and salts thereof in an aqueous solution in a concentration of 0.1 mM to 500 mM;
(B) 0.1 mM to 1 as bicarbonate as a concentration in an aqueous solution
A perfusion solution for ophthalmic surgery, comprising an aqueous solution containing 0.1 mM to 50 mM of 00 mM and (C) glucose as an aqueous solution. (A) a concentration of at least one compound selected from the group consisting of D-3-hydroxybutyric acid and salts thereof in an aqueous solution in a concentration of 0.1 mM to 500 mM;
(B) 0.1 mM to 1 as bicarbonate as a concentration in an aqueous solution
It contains 0.1 mM to 50 mM as a concentration of 00 mM and (C) glucose in an aqueous solution, and has a pH of 6.8 to
8.2. A perfusion solution for ophthalmic surgery, comprising an aqueous solution having an osmotic pressure of 270 to 350 mOsm. (A) at least one compound selected from the group consisting of D-3-hydroxybutyric acid and salts thereof in a concentration of 0.1 mM to 500 mM in an aqueous solution;
(B) 0.1 mM to 1 as bicarbonate as a concentration in an aqueous solution
00 mM, glucose (C) 0.1 mM to 50 mM as a concentration in an aqueous solution, and (D) citric acid and / or an alkali metal salt thereof at a concentration of 0.0
It contains 1 mM to 7.5 mM and has a pH of 6.8 to 8.
2. An ophthalmic perfusion solution preparation comprising an aqueous solution having an osmotic pressure of 270 to 350 mOsm. (A) at least one compound selected from the group consisting of D-3-hydroxybutyric acid and salts thereof in a concentration of 0.1 mM to 500 mM in an aqueous solution;
(B) 0.1 mM to 1 as bicarbonate as a concentration in an aqueous solution
A perfusion solution for ophthalmic surgery, comprising an aqueous solution containing 0.1 mM to 50 mM as a concentration of 00 mM and (E) a phosphate buffer in the aqueous solution. 5. A concentration of at least one compound selected from the group consisting of (A) D-3-hydroxybutyric acid and salts thereof in a concentration of 0.1 mM to 500 mM in an aqueous solution,
(B) 0.1 mM to 1 as bicarbonate as a concentration in an aqueous solution
00 mM, glucose (C) 0.1 mM to 50 mM as a concentration in an aqueous solution, and (D) citric acid and / or an alkali metal salt thereof in an aqueous solution having a concentration of 0.01 m.
A perfusion solution for ophthalmic surgery, comprising an aqueous solution containing M to 7.5 mM and (E) a phosphate buffer in a concentration of 0.1 mM to 50 mM in the aqueous solution.