JPH06271478A - Agent for treating dry eye - Google Patents

Abstract

PURPOSE:To obtain the subject treating agent for ophthalmic field effective for the treatment of dry eye and its relating diseases, etc., such as lowering of lacrimation and Sjogren's syndrome. CONSTITUTION:The objective treating agent contains an interferon (preferably interferon-alpha, etc.) as an active component and a stabilizer (e.g. human serum albumin and sugar), a disinfectant (e.g. benzalkonium chloride), a buffering agent (e.g. boric acid and phosphoric acid), a chelating agent, etc., as arbitrary components.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to the application of interferon in the field of ophthalmology, more specifically to a therapeutic agent for dry eye.

[0002]

[Prior art and its problems] Dry eye is defined as "whether or not there is a keratoconjunctival disorder with a decrease in the amount of tears or abnormal quality of tears" [Yamada, M., et al. ., Ophthalmology, 43, 1289-1293 (1992)], Simple dry eye (lacrima), lacrimal tear, xerophthalmia, Sjögren's syndrome, keratoconjunctivitis sicca, Stevens-Jenson syndrome, pemphigus ophthalmos , Patients with blepharitis, etc. are included in the category of dry eye. In addition, it is said that dry eye associated with cataract surgery and allergic conjunctivitis may be included in the dry eye category. Moreover, in recent years, the development of inspections for dry eye and the increase in interest, and further, VDT (Visual
display terminal) The number of workers and the decrease in tear fluid, which is not abnormal in the whole body, are increasing due to the drying of the room by heating and cooling.

Although there are various causes of dry eye, including unknown ones, it is currently impossible to reactivate the secretion of lacrimal fluid which has been reduced due to the illness. Therefore, a dry eye is diagnosed by performing subjective findings by an interview and objective findings such as the Symmer test, fluorescent dye test and Rose Bencal staining test [Ikuko Toda, Kazuo Tsubota, New Ophthalmology, 8 (7) , 1021-1027 (1991)], an attempt is made to increase the amount of tears stored in the conjunctival sac of a patient, and treatment is currently performed with the goal of alleviating subjective symptoms of the patient.

For the treatment of the above dry eye, conventionally, for example, eye drops such as chondroitin sulfate and methyl cellulose, and oral administration of bromhexine hydrochloride, salivary hormone, bakumondoto and the like have been used, but the effect is not always satisfactory. In cases where it is not possible to do so and is resistant to such treatments and severe cases, punctal coagulation, punctal sutures, punctal plugs, punctal closure such as collagen rods, or eyedrops of artificial tears or contact lenses. , The goggles eye patch is the only method to protect the eyes from dryness [Hamano Taka,
New Ophthalmology, 8 (7), 1029-1035 (1991)].

On the other hand, Interferon (hereinafter abbreviated as IFN) was discovered as an antiviral substance produced by cells stimulated by viruses and the like [Naga
no, Y., et al., CR Soc. Biol., 148, 1700 (1954); Isaacs,
A., et al., Proc. Roy. Soc. B., 147, 258 (1957)].

There are three types of human IFN known to be α, β and γ types. Α type is a cell producing Sendai virus-induced leukocytes, and has a molecular weight of about 15,000 to 21,000.
It consists of 6 amino acids. β-type is produced by double-stranded RNA-induced fibroblasts and has a molecular weight of 22,000.
It consists of 6 amino acids, and the amino acid sequence is approximately 30
% Match. The IFN receptor is also common with α type. The γ-type is a major producer of mitogen-induced T cells, which is composed of 146 amino acids with molecular weights of 20,000 and 25,000.
Unlike types, the receptors for IFN are also different.
Initially, the action of IFN had animal species specificity, and it was difficult to mass-culture human cells, but in the 1980s, the technology for mass-culturing human cells was established, and natural human α-type and β-type Mass production of type IFN was successful with the cell culture method. After that, mass production of recombinant human IFN using gene recombination technology became possible, and clinical application was realized. As a method for mass-producing natural α-type human IFN, a method of using a virus-induced high titer α-type IFN-producing cell from among human acute leukemia cell lines is disclosed in JP-A Nos. 54-98307 and 55-55. -47629 and JP-A-55-62024. Further, as a method for producing human β-type IFN using gene recombination technology, European Published Patent Application 81-
28033, European Published Patent Application 81-321134
, European Published Patent Application 81-34307 and Belgian Patent 81-837397. As a method for producing human γ-type IFN using a gene recombination technique, a method of expression by Gray et al. In Escherichia coli and COS cells [Gray.PW, et al, Nature, 295, 503-508 (1982)]
Is described. Further, in JP-A-55-98118 and JP-A-61-263929, natural γ-IFN is disclosed.
Is described.

[0007] The clinical effects of the above IFN have been reported for renal cancer, multiple myeloma, brain tumor, melanoma, chronic hepatitis B, etc., but in the ophthalmological field, herpes simplex keratitis, etc. Only known for use in viral diseases [Naohiko Tanaka et al., Ophthalmology Clinic, 82 (5),
68-75 (1988)].

[0008]

[Means for Solving the Problems] The inventors of the present invention have been earnestly researching the use of IFN in the ophthalmologic field,
Surprisingly, the effectiveness of IFN has been confirmed against dry eye, which has not been reported yet, and the present invention has been completed here.

That is, the present invention relates to a therapeutic agent for dry eye characterized by containing IFN as an active ingredient.

The therapeutic agent of the present invention has a simple dry eye (hypolacrima) which is conventionally defined as "whether or not there is a keratoconjunctival disorder in a state where the amount of tears is decreased or the quality is abnormal". In addition, various types of dry eye diseases such as lachrymia, xerophthalmia, Sjogren's syndrome, keratoconjunctivitis sicca, Stevens-Jenson syndrome, ocular pemphigus, and eyelid inflammation, cataract surgery and allergic conjunctivitis It is effective for dry eye-like diseases such as dry eye and VDT (decreased tears in workers and dry tears in which there is no abnormality in the whole body caused by dryness of the room due to heating and cooling, etc., especially as eye drops. Particularly, according to the present invention, IFN-IFN-
There is provided a dry eye therapeutic agent for eye drops, which is selected from α, IFN-β and IFN-γ.

IF used as an active ingredient in the therapeutic agent of the present invention
N is not particularly limited as long as it has IFN activity, and includes natural IFN and recombinant IFN produced by gene recombination.
Any of N is included. The IFN includes IFN-α and I
Both FN-β and IFN-γ are included, including all natural and recombinant IFNs. The IFN is also a known culture supernatant containing in vitro-derived human IFN, its purified product, human IFN produced by gene recombination technology, and a synthetic peptide having a partial amino acid sequence. Any of the same IFN-effect substances such as

The production method according to the above-mentioned gene recombination technique can be obtained by an ordinary method, that is, by preparing a vector containing a gene encoding human IFN and its equivalent, and introducing the vector into a host cell for transformation. It can be carried out by culturing the transformed cell, expressing the target substance, and purifying the expression product. Examples of the microorganism used as a host in the above include Escherichia coli and yeast. In the above gene recombination technique, a method of incorporating a gene into an animal cell instead of a microorganism to transform the cell, and expressing the transgene product by culturing the transformed cell is also carried out as a method for producing the IFN. be able to. Examples of animal cells in this method include COS cells, CHO cells and BHK cells.

The gene used in the above gene recombination technique is not particularly limited as long as it is derived from various human cells capable of producing human IFN. For example, HL
-60 from the mRNA isolated from the cell line
Berg method (Okayama H. and Berg, P., Molecular and Cel
lular Biology, 2, 161 (1982)] and the Gubler-Hoffman method [Gubler, V. and Hoffman, BJ, Gene, 25 , 263 (1983)] and the like.

As a method of using Escherichia coli in the above gene recombination technique, the method of Pennica et al. [Pennic
a, D., Nature, 312, 724 (1984)]. In addition, human IFN and a part of human IFN can be modified or a human IFN derivative can be produced by utilizing a known gene recombination technique.
It can be carried out by a conventional method in this field.

The dry eye therapeutic agent of the present invention can be generally used in the form of a general pharmaceutical preparation. Various formulations can be selected according to the therapeutic purpose, and typical examples thereof include tablets, pills, powders, solutions, suspensions, capsules, suppositories, and injections (solutions, suspensions). Agent), ointment,
Examples thereof include eye drops, and eye drops are particularly preferable.

In clinical use of the therapeutic agent for dry eye of the present invention, IFN (IFN-α, IFN-β) as an active ingredient is used.
Or, the amount of IFN-γ) used is usually 1 × 1 per day for an adult.
0 ⁰ Units / eye to 1 × 10 ⁸ units / eye about, preferably 1
It is desirable to set the range of approximately 10 ⁴ units / eye to 200 × 10 ⁴ units / eye.

Further, the method of using the therapeutic agent of the present invention is usually 1 to 6 drops / eye for an adult, preferably 2 to 3 drops / eye once a day.
It is possible to employ a method of instilling each eye in 15 to 15 times or in 5 to 6 times a day. The above-mentioned applied amount can be appropriately changed depending on the age, symptoms, etc. of the patient.

The ophthalmic preparation containing the IFN of the present invention as an active ingredient is a variety of components that can be used as usual ophthalmic preparations, for example, stabilizers, bactericides, buffers, isotonic agents,
It can be prepared by appropriately using a chelating agent, a pH adjusting agent, a surfactant and the like.

Examples of stabilizers for the above-mentioned ophthalmic preparations include human serum albumin, ordinary L-amino acids, saccharides, cellulose derivatives and the like, which can be used alone or in combination with a surfactant or the like. In particular, this combination may be able to further improve the stability of the active ingredient IFN.

The L-amino acid is not particularly limited and may be any of glycine, cystine, glutamic acid and the like.

The sugar is not particularly limited, and examples thereof include monosaccharides such as glucose, mannose, galactose and fructose, sugar alcohols such as mannitol, inositol and xylitol, disaccharides such as sucrose, maltose and lactose, dextran and hydroxypropyl. Polysaccharides such as starch, chondroitin sulfate, hyaluronic acid and the like and their derivatives can be used.

The surfactant is not particularly limited, and both ionic and nonionic surfactants can be used. Examples thereof include polyoxyethylene glycol sorbitan alkyl ester type, polyoxyethylene alkyl ether type,
Examples thereof include sorbitan monoacyl ester type and fatty acid glyceride type.

The cellulose derivative is not particularly limited, and examples thereof include methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose and the like. Among the above cellulose derivatives, cellulose such as hydroxypropyl cellulose used as a component of artificial tears can be effectively blended particularly in the dry eye therapeutic agent of the present invention.

The amount of the saccharide added is about 0.000 per 1 μg of IFN itself or the derivative (IFN active substance).
It is suitable that the amount is about 1 mg or more, preferably about 0.01 to 10 mg. The amount of surfactant added is
It is suitable that the amount is about 0.00001 mg or more, preferably about 0.0001 to 0.01 mg per 1 μg of each of the above-mentioned active substances. The amount of human serum albumin added is appropriately about 0.0001 mg or more, preferably about 0.001 to 0.1 mg per 1 μg of each of the above-mentioned active substances. Amino acid is 1 μg of each of the above active substances
About 0.001 to 10 mg per unit is suitable. Further, the addition amount of the cellulose derivative is appropriately in the range of about 0.00001 mg or more, preferably about 0.001 to 0.1 mg per 1 μg of each of the above active substances.

The amount of IFN contained in the pharmaceutical preparation of the present invention as an active ingredient is not particularly limited and may be appropriately selected from a wide range. Usually, the total amount of active ingredients is about 0.00.
001 to 70% by weight, preferably 0.0001 to 5% by weight is suitable.

The bactericidal component used as a component of the ophthalmic formulation of the present invention may be any bactericidal agent which can be usually used in the ophthalmic field, for example, chlorhexidine salt, benzalkonium salt, paraoxybenzoic acid ester. , Chlorbutanol, benzethonium salt, cetylpyridinium salt, thimerosal, hydrogen peroxide and the like, more specifically, chlorhexidine salts such as chlorhexidine digluconate and benzalkonium salts such as benzalkonium chloride can be advantageously used. In the case of the above benzalkonium chloride, 0.001 w. It is preferably used at about v% to 0.05 w / v%.

The above bactericides may be used alone or in combination of two or more which do not react with each other.
In consideration of the pathological condition of dry eye, which is the target of the ophthalmic formulation of the present invention, an ophthalmic formulation containing the above-mentioned minimum amount of the bactericide or not containing it at all can be appropriately adjusted. Further, the ophthalmic formulation of the present invention may be used in an appropriate dosage form such as a liquid or an eye ointment for the purpose of treating dry eye, and for this purpose, an appropriate buffer, isotonicity agent, chelating agent. A medicinal ingredient that can be used in various ophthalmic areas such as, for example, artificial tears and the like may be contained.

Examples of the buffering agent include boric acid, phosphoric acid,
Examples thereof include acetic acid, citric acid, ε-aminocaproic acid, glutamic acid, and / or their corresponding salts (for example, their sodium salts, potassium salts, calcium salts, alkali metal salts such as magnesium salts, alkaline earth metal salts) and the like. . Examples of the isotonicity agent include sodium chloride, potassium chloride, sugars, glycerin and the like. Further, as the chelating agent, for example, sodium edetate, citric acid, etc. can be appropriately used. The chelating agent can also function as a stabilizer.

Further, the ophthalmic preparation of the present invention has a liquidity that can be used for ophthalmology, for example, it is added with the buffering agent exemplified above.
It is preferably adjusted to about H5 to pH8.

In the use of the ophthalmic preparation of the present invention, in addition to the solution ophthalmic preparation, the ophthalmic preparation is freeze-dried into a storable state and then water for use, physiological saline and / or Alternatively, it can be used after being dissolved in a buffer solution containing artificial tears and adjusted to a concentration suitable for eye drops.

The suitable concentration of IFN of the active ingredient of the present invention is generally 2 × 10 ⁴ IU / ml to 200 × 10.
^It is preferable to adjust the concentration to ⁴ IU / ml before use.

[0032]

INDUSTRIAL APPLICABILITY The therapeutic agent of the present invention is a dry eye which is defined as "eye drops may be present or not with or without keratoconjunctival disorder" in a state where the amount of tears is abnormal or abnormal. Useful for dry eye related diseases.

[0033]

[Examples] Formulation examples and test examples of the therapeutic agent of the present invention are given below.

[0034]

[Formulation Example 1] α-interferon 5.0 × 10 ⁶ IU / ml HSA (human serum albumin) 5 mg / ml Cysteine 0.1 mg / ml The above components were mixed at the above concentrations, and the mixture was filtered ( 0.22 μm membrane filter was used), and then the filtrate was aseptically dispensed in 1 ml portions into a polyethylene container or a glass container and freeze-dried to prepare the therapeutic agent of the present invention in the form of eye drops.

The preparation is used by dissolving it in physiological saline containing a suitable preservative at the time of use, or by dissolving it in artificial tears.

[0036]

[Formulation Example 2] α-interferon 5.0 × 10 ⁶ IU / ml HSA (human serum albumin) 5 mg / ml Hydroxypropylcellulose 1 mg / ml The above components were mixed at the above concentrations, and the mixture was filtered ( 0.22 μm membrane filter was used), and then the filtrate was aseptically dispensed in 1 ml portions into a polyethylene container or a glass container and freeze-dried to prepare the therapeutic agent of the present invention in the form of eye drops.

The preparation is used by dissolving it in physiological saline containing an appropriate preservative at the time of use.

[0038]

[Formulation Example 3] δ-interferon 5.0 × 10 ⁶ IU / ml HSA (human serum albumin) 5 mg / ml cysteine 0.1 mg / ml Tween 80 0.01 mg / ml Each of the above components was adjusted to the above concentration at 0. 0.01 M citric acid-sodium citrate buffer (pH 6.0) was added and mixed, and the mixture was filtered (using a 0.22 μm membrane filter), and then 1 ml of the filter was aseptically divided into polyethylene containers or glass containers. It was poured and freeze-dried to prepare the therapeutic agent of the present invention in the form of eye drops.

The preparation is used by dissolving it in physiological saline at the time of use or by dissolving it in artificial tears.

[0040]

[Formulation Example 4] β-interferon 5.0 × 10 ⁶ IU / ml HSA (human serum albumin) 5 mg / ml Cysteine 0.1 mg / ml Tween 80 0.01 mg / ml Dextran 40 15 mg / ml To 0.01M citric acid-sodium citrate buffer (pH 6.0) and mixed, and after filtering the mixture (using a 0.22 μm membrane filter), aseptically filter each 1 ml polyethylene container. Alternatively, it was dispensed in a glass container and freeze-dried to prepare the therapeutic agent of the present invention in the form of an eye drop preparation.

The preparation 4 is used by dissolving it in physiological saline containing an appropriate preservative at the time of use, or by dissolving it in an artificial lacrimal fluid.

[0042]

[Formulation Example 5] α-interferon 5.0 × 10 ⁶ IU / ml HSA (human serum albumin) 5 mg / ml The above components were mixed at the above concentrations, and the mixture was filtered (using a 0.22 μm membrane filter). After that, 1 ml each of the filtrate was aseptically dispensed into a polyethylene container or a glass container, and freeze-dried to prepare the therapeutic agent of the present invention in the form of eye drops.

The preparation 5 is used by dissolving it in physiological saline containing a suitable preservative at the time of use, or by dissolving it in an artificial lacrimal fluid.

[0044]

[Test Example 1] OPC-18 (α-type interferon: 1
5 million units in a vial containing SoftSantia (component: 0.1% potassium chloride, 0.4% sodium chloride;
When dissolved in 2.5 ml of Santen Pharmaceutical Co., Ltd., it was dissolved to prepare 2 million units / ml (1 eye drop, 100,000 units).

A 46-year-old (female) patient with moderate dry eye was instilled with the above-prepared eyedrops in each patient's eye at 2 to 3 drops, 5 to 6 times a day, and continued for 4 weeks. Was administered.

Before administration, after 4 weeks from the start of administration and after completion of administration 4
The effects of the above eye drops (the present invention) were examined on subjective symptoms of patients after week and subjective findings of dry eye in a vital stain test.

As a result, the subjective symptoms were written according to the patient's self-confidence regarding the following observation items and the degree of the symptoms (judgment stage 0 to 5), and the total score was used as the subjective symptom score.

<Observation Items> Eyes tired, eyes feel dry, eyes tingle, eyes tingle, eyes hurt, glare, glare, bright eyes, red eyes, greasy eyes, heavy eyes, discomfort in the morning, I can't open my eyes I have tears I have itchy eyes and my eyes are hot.

1 ... Sometimes this symptom is present. There is no hindrance to everyday life.

2 ... Not so bad, but there is always this symptom. There is no hindrance to daily life.

3 ... I am always worried and anxious. I can manage my daily life normally.

4 ... Symptoms are severely painful, but I can manage to endure it. It has a great influence on daily life.

5 ... Symptoms are so severe and unbearable. I can't live everyday.

(The term "daily life" as used herein refers to daily activities performed by the subject. For example, reading, driving, computer work, watching TV, etc.) The vital staining test for objective findings is different. , 3 to keratoconjunctiva
It is divided into 7 areas, and the Rose Bengal staining is scored from 0 to unstained depending on the degree of staining, mild 1, moderate 2,
It is classified into Severity 3 and Fluorescein staining is also classified according to the degree of staining, from 0 for unstained to Mild 1, Moderate 2 and Severe 3, and the total score is rose. Shown as Bengal score (RB) and Fluorescein score (F).

Changes in subjective symptoms and changes in vital staining test before and after administration of the therapeutic agent of the present invention are shown below.

Date 7/6 8/4 9/14 Subjective symptoms 36 28 34 RB (right eye / left eye) 10/10 7/6 10/10 F (right eye / left eye) 9/9 4/4 9/9 The above effect continued until around 9/12.

From the above results, the therapeutic agent of the present invention (eye drops)
Was confirmed to have an excellent effect of improving subjective symptoms of patients with dry eye and an excellent improving effect in a dyeing test, and it could be determined to be effective as a dry eye therapeutic agent for dry eye patients.

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[Procedure amendment]

[Submission date] April 26, 1993

[Procedure Amendment 1]

[Document name to be amended] Statement

[Correction target item name] 0038

[Correction method] Change

[Correction content]

[0038]

[Formulation Example 3] γ -interferon 5.0 × 10 ⁶ IU / ml HSA (human serum albumin) 5 mg / ml Cysteine 0.1 mg / ml Tween 80 0.01 mg / ml Each of the above components was adjusted to the above concentration at 0. 0.01 M citric acid-sodium citrate buffer (pH 6.0) was added and mixed, and the mixture was filtered (using a 0.22 μm membrane filter), and then 1 ml of the filter was aseptically divided into polyethylene containers or glass containers. It was poured and freeze-dried to prepare the therapeutic agent of the present invention in the form of eye drops.

Claims (4)

[Claims] 1. A therapeutic agent for dry eye, which comprises interferon as an active ingredient. 2. The dry eye is selected from the group consisting of hypolacrima, lacrimal tear, xerophthalmia, Sjogren's syndrome, keratoconjunctivitis sicca, Stevens-Jenson syndrome, and ocular pemphigus. The dry eye therapeutic agent described. 3. Interferon is interferon-
The dry eye therapeutic agent according to claim 1, which is α. 4. An eye drop, which is characterized in that
The therapeutic agent for dry eye according to.