WO2005003111A1

WO2005003111A1 - Preventive or remedy for viral conjunctivitis

Info

Publication number: WO2005003111A1
Application number: PCT/JP2004/009701
Authority: WO
Inventors: Jun Hiroi; Eisaku Tsujii; Tatsuo Suzutani; Hisatoshi Kaneko; Akihiko Miyatake
Original assignee: Astellas Pharma Inc.
Priority date: 2003-07-04
Filing date: 2004-07-01
Publication date: 2005-01-13

Abstract

It is intended to provide an antiviral agent against adenoviruses, in particular, adenovirus type 8, type 19 or type 37 and a preventive or a remedy for viral conjunctivitis induced by adenoviruses which are characterized by containing cromoglicic acid or its salt as the active ingredient; and a method of preventing or treating influenza which comprises administering the same into humans or animals.

Description

Prophylactic or therapeutic agent for viral conjunctivitis

The present invention relates to an antiviral agent for adenovirus. More specifically, the present invention relates to a prophylactic or therapeutic agent for viral conjunctivitis, comprising cromoglycic acid or a non-toxic salt thereof as an active ingredient, and a virus comprising administering these to humans or animals. And a method for preventing or treating conjunctivitis conjunctivitis. Background art

Adenovirus is responsible for 80 to 90% of viral conjunctivitis, and it is said to affect about 100,000 people a year in Japan.ル Since conjunctivitis is a highly contagious infectious disease, daily life is restricted and forced to leave school and workplaces if it becomes ill. Furthermore, hospital-acquired infections caused by the 7 denovirus have recently become an important social issue in public health, and have become an economic and ethical issue in hospitals.

However, as various infectious diseases have been overcome, there is no established drug that is effective and highly safe for viral conjunctivitis. Observed coping therapy is mainstream. Therefore, development of a drug having a reliable antiviral effect against adenovirus causing viral conjunctivitis is expected.

Human adenovirus was discovered by Rowe and Hillman. As common feature of adenoviridae (Adenoviridae), viral particles without a positive 2 0 tetrahedron (diameter 7 0 to 9 0 nm), the genome is a double-stranded DNA (molecular weight ^{2 0~ 2 5 X 1 0 6} ) It has an inverted repeat sequence of 100 to 160 bases at both ends. Currently, serotypes range to 51, with the addition of a new strain isolated from AIDS patients. It is divided into six groups (subgenus) based on indicators such as genome homology and recombination ability within the group. Minute of adenovirus The types are shown in Table 1.

Table 1 Classification of adenovirus

Adenovirus-related diseases are widespread, but the serotypes that cause conjunctivitis are

Types 3, 4, 7, 8, 11, 19 and 37 have been reported. In the subgenus, the subgenus B (3, 7, 11), E (4) and D (8, 19, 37) fall into this category (Hideo Uchio, New Ophthalmology, 20 (3), p289-295 (2003 )). Among them, adenovirus types 8, 19 and 37 are responsible for the outbreak of conjunctivitis in Asian countries including Japan, especially in East and Southeast Asia (K. Aoki et al., Br. J. Ophthalmol. ., 66 p776-780 (1982)). In addition, adenovirus types 8, 19 and 37 are well known epidemiological causes of nosocomial infections (J.A. Jernigan et al., J. Infect. Dis., 167 pl307-1313 (1993)).

On the other hand, sodium cromoglycate is used clinically as an antiallergic agent for bronchial asthma and allergic rhinitis and allergic conjunctivitis, and its side effects are minimal, such as cough sometimes seen when inhaling powdered formulations It is clinically recognized as a very safe drug. Sodium cromoglycate has not been reported to have any serious side effects, despite its long history of clinical administration, and most notably, it has established its effects and safety in children. To date, there is no finding that sodium lipodium molycdate is effective for viral conjunctivitis. For antiviral effects against adenovirus, see Penttinen et al. This report describes the antiviral activity of sodium cromoglycate.This report describes experiments using viruses different from adenoviruses that cause conjunctivitis (Adenovirus 5, Echovirus 7, Herpes simplex, and Semi iki Forest virus). Sodium cromoglycate does not affect the growth of the virus but inhibits the cell damage caused by the virus (K. Penttinen et al., Brit. Med. J., I, 6053, p82 (1977)

The present invention has been made in view of the current situation in the prior art, and has as its object to provide an antiviral agent for adenovirus which is highly safe for the human body, and particularly for safety for children. The purpose is to provide existing drugs with established sexual properties as preventive or therapeutic agents for viral conjunctivitis. Disclosure of the invention

The present inventors have found that sodium cromoglycate is effective for adenoviruses that cause conjunctivitis in the process of examining the antiviral effect of sodium cromoglycate, particularly for adenoviruses 8, 19 and 37. The present invention has been completed. Therefore, according to the present invention, there is provided an agent for preventing or treating viral conjunctivitis, comprising as an active ingredient cromoglycic acid or a salt thereof as shown in the following [1] to [S], Also provided is a method for preventing or treating diarrheal conjunctivitis comprising administering these to humans or animals.

[1] An antiviral agent against adenovirus, comprising cromoglycic acid or a salt thereof as an active ingredient.

[2] An antiviral agent against adenovirus type 8, type 19 or type 37, comprising cromoglycic acid or a salt thereof as an active ingredient.

[3] An agent for preventing or treating viral conjunctivitis, comprising cromoglycic acid or a salt thereof as an active ingredient.

[4] A prophylactic or therapeutic agent for viral conjunctivitis caused by adenovirus type 8, type 19 or type 37 infection, comprising cromoglycic acid or a salt thereof as an active ingredient. [5] An agent for preventing the onset of clinical symptoms of diluent conjunctivitis or alleviating the symptoms, comprising cromoglycic acid or a salt thereof as an active ingredient.

[6] A method for preventing or treating viral conjunctivitis, comprising administering an effective amount of cromoglycic acid or a salt thereof to a human or animal.

[7] A method for preventing the onset or alleviating the clinical symptoms of viral conjunctivitis, comprising administering an effective amount of cromoglycic acid or a salt thereof to a human or animal.

[8] The agent or the method according to any one of claims 1 to 7, wherein the cromoglycic acid or a salt thereof is sodium mogalycate salt. BEST MODE FOR CARRYING OUT THE INVENTION

Viral conjunctivitis, which is the subject of the present invention, is initiated by the attachment of adenovirus, particularly adenovirus type 8, 19 or 37, to a receptor present in epithelial cells of the conjunctiva. It is a disease. After infection, the virus transfers into the cells and multiplies in the nucleus. When the virus particles fill the nucleus, the virus releases virus particles and the infection spreads.

The cromoglycic acid used in the present invention is a known compound 5,5 '-(2-hydroxytrimethylenedioxy) bis (4-oxo-1H-1-benzopyran-12-carboxylic acid).

Non-toxic salts of cromoglycic acid include, for example, salts with alkali metals such as sodium or potassium, salts with alkaline earth metals such as calcium or magnesium, ammonium salts, and quaternary ammonium salts. Salts may be mentioned, and among these, sodium cromoglycate is particularly preferred.

The agent for preventing or treating viral conjunctivitis of the present invention is preferably used as a topical preparation containing the active ingredient cromoglycic acid or a nontoxic salt thereof, for example, as an eye drop. The content (concentration) of sodium cromoglycate in the topical preparation of the present invention is 0.5 to 5%, preferably 1 to 2%.

Next, a method for producing the topical preparation of the present invention will be described. For example, to prepare eye drops, sodium cromoglycate is dissolved in purified water or sterilized purified water by stirring. These preparations may optionally contain a tonicity agent such as sodium chloride. ), Buffers (eg, boric acid, sodium monohydrogen phosphate, sodium dihydrogen phosphate, etc.), preservatives (eg, benzalkonium chloride, etc.), thickeners (eg, carboxyl vinyl polymer, etc.), stabilizers (eg, Commonly used additives such as sodium detorate, a refreshing agent (eg, 1-menthol), dl-camphor, d-borneol, fennel oil, heart oil, heart water can be added. .

The thus obtained topical preparation of the present invention is administered once to several times a day depending on the condition of the patient.

Cromoglycic acid or a salt thereof may be used in combination with other drugs, antihistamines (eg, diphenhydramine hydrochloride, chlorpheniramine maleate, promethazine, diphenirarine hydrochloride, clemastine fumarate, etc.), vasoconstrictors (eg, norfuenefrin hydrochloride, It can also be used in combination with feneryphrine hydrochloride, naphazoline hydrochloride, gyrometazoline, mitodrine hydrochloride, methoxamine hydrochloride, tetrahydrooctrozoline nitrate), steroid anti-inflammatory agents, antibiotics and the like. The individual components of such combinations may be administered sequentially or simultaneously in separate or combined pharmaceutical formulations.

Eye drops containing cromoglycic acid or its salts that are effective for the prevention or treatment of viral conjunctivitis are administered once or several times daily, depending on the type of human or animal being treated and the condition of the patient . When the present invention is administered as a prophylactic agent for viral conjunctivitis, it can be started during the viral conjunctivitis epidemic season, and can be carried out with increasing or decreasing as needed. In other words, when viral conjunctivitis epidemic is imminent, or before or after possible infection (such as swimming in a pool) or before the onset of viral conjunctivitis. Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to these Examples.

Hereinafter, the preparation for topical administration of the present invention will be described with reference to Examples. In addition, 2% sodium cromoglycate ophthalmic solution is marketed as Intal (R) ophthalmic solution and Intal (R) ophthalmic solution UD. (Fujisawa Pharmaceutical Co., Ltd.)

Example 1 Sodium cromoglycate, d1-chlorpheniramine maleate, tifazoline hydrochloride and sodium edetate were dissolved in purified water, benzonalconium chloride was added thereto, and the mixture was stirred. A prescribed nasal drop was obtained.

Sodium cromoglycate 1 g

d 1 Chlorophyle maleate 0.25 g

Nafazoline hydrochloride 0.025 g

Sodium edetrate 0.01 g

Benzalkonium hydrochloride 0.01 Total 100ml

Example 2

Sodium cromoglycate, d1-chlorpheniramine maleate, sodium edetate, benzalkonium chloride, and sodium chloride were added to sterile purified water, stirred, dissolved and sterile-filtered to obtain eye drops of the following formulation.

Sodium cromoglycate 2 g

d 1—Chlorfe maleate 0.4 g

Sodium edetrate 0.01 g

Benzalkonium hydrochloride 0.01 g

NaCl chloride 0.3 g Total volume 00ml

Example 3

Sodium cromoglycate, d1-chlorpheniramine maleate, toluene, benzalkonium chloride, borax, and boric acid were added to sterile purified water, stirred and dissolved, and aseptically filtered to obtain eye drops of the following formulation.

Sodium cromoglycate 1 g

d 1—Chlorpheniramine maleate 0 015 g

1 1 Men! 0 006 g

Benzalkonium chloride 0 002 g Borax 0.01 g

Boric acid 1.8 g

100ml

Example 4

Sodium cromoglycate, diphenhydramine hydrochloride, 1-menthol, benzalkonidum, borax, and boric acid were added to sterilized purified water, stirred and dissolved, and aseptically filtered to obtain eye drops of the following formulation.

Sodium cromoglycate l g

Diphenhydramine hydrochloride 0.02 g

1 menthol 0.006 g

Benzalkonium chloride 0.002 g

Borax 0.01 g

Boric acid 1.8 g

Appropriate amount of sterilized purified water

Total volume 100ml

Test example

Effect of sodium cromoglycate on adenovirus infection (in vitro test) The degree of cytotoxicity due to virus infection in vitro was evaluated by the enzyme in intracellular mitochondria using MTT (3- (4,5-dimethyl-2-thiazolyl ) -2,5-Diphenyl-2H-tetrazolium · Br) was measured by measuring the amount of MTT formazan produced by reduction at 570 nm to quantitatively determine the number of viable cells.

I- Test material

Adenoviruses the three strains as shown in Table 2, the amount of virus, respectively 100MTT ID _5. It was used after dilution.

table 1 Adenovirus serotype derived

Type 8 clinical isolate

Type 19 clinical isolate

Type 37 clinical isolate The ID (Infectious dose) indicating the amount of virus infection is a unit of the amount of virus required for infection of host cells, and was measured and determined as follows.

Save virus solution, diluted to 2 ⁸ times with MEM medium, microplates (96 flat bottom Plate, co-made-learning Co.) HEL which had been cultured in (Human embryo lung fibrobla st, human embryonic lung cells) the virus dilutions 50 ^ 1 was added 6 days C0 ₂ incubator beta - (. ID ₅₎ was cultured in performs MTT staining, viral load surviving cells amount after viral infection is 50 percent each strain Was determined. 100 times the virus concentration was used for the following tests.

The sodium cromoglycate solution was dissolved in a MEM medium to a concentration of 20 mg / ml and sterilized with a 0.2 m filter. This solution was transferred in a 96-well plate into which 200 1 of MEM medium was dispensed, and a test dilution of 5-fold dilution of sodium cromoglycate was prepared.

I I. Test method

Using a 96-well culture microplate (manufactured by Koingen Co., Ltd.), prepare 2 x 10 ⁴ HEL cells in MEM medium 100 containing 10% fetal bovine serum per 1-well. The solution was inoculated. After culturing for 16 to 24 hours in the presence of 37 ° (5% C 存在₂ ), it was confirmed under a microscope that a monolayer was formed on the aerial surface.

The solution of each virus indicated that the culture supernatant up to a concentration 2 QmgZml or al the suction advance another plate to the 5-fold serial dilutions of sodium cromoglycate solution to ₀ more Table 1 was added in each IOOI the 100MTT ID _5. Diluted, and the culture medium and uninfected control media 100 l containing no virus added to the different cells as 37 ° C, 5% concentration C0 ₂ 6 days in the presence of containing virus.

After completion of the culture, the 4-hour culture supernatant after sucked by the addition of 5MgZml MTT solution 1 OI 37 ° CC_〇 ₂ incubator beta within one, isopropylidene containing 0. 05N HC 1 After adding formic acid 1001 and mixing to completely dissolve the formazone, measure the absorbance of each well using a microplate reader (TECAN, measurement wavelength 570 nm, control wavelength 660 nm) to determine the infection. The amount of remaining viable cells was measured.

III. Test results: Suppression of sodium cromoglycate against cell death due to growth of each serotype adenovirus, calculated assuming that the absorbance of uninfected cells is 100% survival and the absorbance of drug-free wells is 0% inhibition. EC _{5 of} action. And the CC ₅ 0 is an indication of the cytotoxicity to uninfected cells are shown in Table 2. Sodium cromoglycate suppressed the cytotoxicity of 0.85 to 1.53 mgZml in the conjunctivitis clinical isolates tested, due to the propagation of serotype 8, 19, and 37 adenoviruses. However, CC ₅ for HEL cells, a host cell. Was not observed even at 1 Omg / m1. Table 2

Industrial applicability

According to the present invention, an anti-adenovirus agent highly safe for the human body, more specifically, a prophylactic or therapeutic agent for viral conjunctivitis, which comprises cromoglycic acid or a salt thereof as an active ingredient, and A method for preventing or treating viral conjunctivitis comprising administering these to humans or animals is provided.

Claims

The scope of the claims

1. An antiviral agent against adenovirus, comprising cromoglycic acid or a salt thereof as an active ingredient. '

2. An antiviral agent against adenovirus type 8, type 19 or type 37, characterized by containing cromoglycic acid or a salt thereof as an active ingredient.

3. An agent for preventing or treating diarrheal conjunctivitis, comprising cromoglycic acid or a salt thereof as an active ingredient.

4. An agent for preventing or treating viral conjunctivitis caused by infection with adenovirus type 8, 19, or 37, comprising cromoglycic acid or a salt thereof as an active ingredient.

5. An agent for preventing the onset of symptomatic conjunctivitis or for alleviating the symptoms thereof, comprising cromoglycic acid or a salt thereof as an active ingredient.

6. A method for preventing or treating viral conjunctivitis, comprising administering an effective amount of cromoglycic acid or a salt thereof to a human or animal.

7. A method for preventing the onset or alleviating the clinical symptoms of virus '14 conjunctivitis, comprising administering an effective amount of cromoglycic acid or a salt thereof to a human or animal.

8. The agent or method according to any one of 1 to 7, wherein the clomoglycic acid or a salt thereof is sodium cromoglycate.