JPH039883B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an external preparation for treating dyskeratosis of the skin and mucous membranes containing interferon as an active ingredient.

Lesions observed on the skin and mucous membranes include stomatitis caused by infections such as herpes simplex virus and herpes zoster virus, viral diseases such as herpes simplex and shingles, and pathogens such as candidiasis caused by fungi. There are infections caused by sexually transmitted microorganisms and keratinization abnormalities whose etiology is completely unknown, such as lichen planus and leukoplakia. Among these skin and mucous membrane lesions, antiviral agents, antibiotics, antifungal agents, anti-inflammatory agents, and the like are known to be effective in treating viral diseases and infectious diseases whose etiology is clear. However, as for the treatment of abnormal keratosis such as lichen planus and leukoplakia, the etiology of which is unknown, currently only vitamin preparations and corticosteroids are administered, and no effective means have been found.

Lichen planus is a chronic inflammation accompanied by abnormal keratosis found in the skin and mucous membranes, such as the mucous membranes in the oral cavity.It is common in middle-aged and older people, especially women, and when erosion occurs in the oral cavity, it stains when eating or comes into contact with the skin. It is painful and there are several types, but the ulcer type and blister type are said to be more likely to turn into cancer. Leukoplakia is a white spot-like lesion that occurs due to hyperkeratosis of the mucosal epithelial layer of the oral mucosa, etc., and is said to occur more often in middle-aged and older men, and can turn into cancer.

Although abnormal keratosis of the skin and mucous membranes such as lichen planus and leukoplakia are a type of precancerous disease, there is no effective treatment because the etiology is unknown.
Its appearance is strongly desired.

In view of these circumstances, the present inventors have conducted extensive research to find an effective treatment for abnormal keratosis of the skin and mucous membranes such as lichen planus and leukoplakia. It was found that it has a remarkable therapeutic effect on abnormalities.

In other words, since interferon was discovered as a virus growth inhibitor produced by cells, subsequent research has revealed various pleiotropic biological effects, and recent technological advances have led to interferon being derived from human cells and humans. It has become possible to mass-produce and purify interferon derived from microorganisms with interferon gene integration suitable for clinical application, and to treat herpetic keratitis, hepatitis B, viral warts,
Clinical applications are underway for brain tumors, skin melanoma, etc.

However, interferon is an extremely unstable substance, and its activity is significantly lost when subjected to temperature (high temperature) or physical pressure, especially when purified to the extent that it can be used clinically. Therefore, currently,
Interferon is clinically applied primarily as a freeze-dried product, and before use, it is dissolved in physiological saline, purified water, etc., and administered in the form of an injection or eye drops.

However, the present inventors surprisingly found that when interferon was made to coexist with a trivalent or higher valent sugar alcohol and an organic acid buffer, the stability of interferon was significantly improved. We found that a topical preparation has a significant therapeutic effect on abnormal keratosis of the skin and mucous membranes of unknown etiology, such as lichen planus and leukoplakia. So far, there has been no pharmacological or clinical report showing that interferon is effective against these abnormal keratosis.

The present invention has been completed based on this knowledge, and is an external preparation for treating abnormal keratosis of the skin and mucous membranes, which contains interferon as an active ingredient, and contains a trivalent or higher valent sugar alcohol and an organic acid buffer. It provides: According to the present invention, it is possible to effectively treat abnormal keratosis of the skin and mucous membranes of unknown etiology, such as lichen planus and leukoplakia, simply by applying interferon externally to the affected area. .

The interferon used as the active ingredient of the external preparation of the present invention may be of any human origin, for example, it may be derived from human leukocytes or normal diploid cells, or it may be derived from human leukocytes or normal diploid cells, or it may be derived from human interferon genes incorporated using recombinant DNA techniques. Those derived from microorganisms are used. The blending amount is not particularly limited and can be selected appropriately depending on the actual dosage form as an external preparation, but from the viewpoint of effectiveness, it is generally 1×
It is preferable to mix interferon having a potency of 10 ⁵ international units or more in a proportion of 1×4 ⁴ international units or more per 100 g of the composition.

Examples of the trihydric or higher sugar alcohol to be used include glycerin, erythritol, arabitol, xylitol, sorbitol, mannitol, etc. These may be used alone or in combination of two or more types, and from the viewpoint of stability of interferon, The blending amount is 15% (weight%, same hereinafter) or more of the total external preparation as a sugar alcohol, preferably,
It is desirable to set it to 25-70%.

The organic acid buffer may be a conventional buffer consisting of an organic acid or its salt, such as monosodium citrate.
Citrate buffers such as disodium citrate mixtures, citric acid-trisodium citrate mixtures, citric acid-monosodium citrate mixtures, succinic acid-monosodium citrate mixtures, succinic acid-monosodium citrate mixtures,
Succinate buffers such as sodium hydroxide mixtures, succinic acid-disodium succinate mixtures, tartrate buffers such as tartaric acid-sodium tartrate mixtures, tartaric acid-potassium tartrate mixtures, tartaric acid-sodium hydroxide mixtures, fumaric acid - fumarate buffers, such as monosodium fumarate mixtures, fumaric acid-disodium fumarate mixtures, monosodium fumarate-disodium fumarate mixtures,
Gluconic acid-sodium gluconate mixture, gluconic acid-sodium hydroxide mixture, gluconic acid-
Gluconate buffers such as potassium gluconate mixtures, oxalic acid-sodium oxalate mixtures,
Oxalic acid-sodium hydroxide mixture, oxalic acid-
Oxalate buffers such as potassium oxalate mixtures, lactic acid-sodium lactate mixtures, lactic acid-sodium hydroxide mixtures, lactate buffers such as lactic acid-potassium lactate mixtures, acetic acid-sodium acetate mixtures, acetic acid-sodium hydroxide Examples include acetate buffers such as mixtures. From the viewpoint of stability of interferon, these organic acid buffers should be used in an amount of 0.01 mol/Kg or more, preferably 0.1 mol/Kg, based on the entire external preparation.
It is desirable to add mol/kg or more and adjust the pH of the external preparation to 3 to 6.

The external preparations of the present invention include ointments, paste preparations, gel preparations,
It can be made into a dosage form suitable for external use such as a spray or a liquid. These can be manufactured according to conventional formulation techniques, and interferon may be added in the final step of manufacturing. Other ingredients are not particularly limited as long as they do not affect the stability of interferon, and any of those commonly used in this type of preparation may be used. In particular, when preparing a solid preparation, it is preferable to use carboxymethylcellulose as a binder in view of the stability of interferon.

In order to use the external preparation of the present invention for the treatment of abnormal keratosis of the skin or mucous membranes such as lichen planus or leukoplakia, the external preparation may be applied directly to the affected area by coating or spraying. Although not particularly limited, in order to prevent interferon from being removed from the affected area and to promote the absorption of copper and enhance the therapeutic effect, the area to which the topical preparation has been applied should be covered with a bandage or bandage. It is preferable to cover it with etc. In particular,
When applying to the affected area on the oral mucosa, an oral mucosal adhesive bandage (for example, a
11299) etc., the therapeutic effect can be enhanced.

The dosage is usually 10 to 10 per dosage unit (one time).
It is preferable that 10 ⁸ international units of interferon can be applied, for example, a gel containing interferon of 10 ⁴ to 10 ⁹ international units/100 g for external use, or 10 ⁵ to 10 ¹⁰ international units/100 g for external use. When using an ointment containing interferon, the dose should be 0.01~
Just apply 10g to the affected area with your fingers, spatula, etc. two to three times a day.

Next, the present invention will be explained in more detail with reference to Examples and Test Examples. The interferon used in the following examples and test examples is obtained from human foreskin-derived fibroblasts, and has passed the standards for clinical application set by the National Institute of Preventive Health in terms of purity, production volume, etc. and its safety has been confirmed in the preclinical tests required by the Ministry of Health and Welfare.

Example 1 Gel base formulation Ingredients % Sodium lauryl sulfate 0.2 Carboxymethyl cellulose 2.0 Glycerin 40.0 0.4 mol/Citrate buffer (PH4.5) 25.0 Purified water Residual Each component was mixed to obtain a base, and per 100g of it,
Interferon was mixed in a proportion of 1×10 ⁷ international units to obtain a gel for external use.

Example 2 Gel base formulation Ingredients % Carboxymethyl cellulose 2.0 Glycerin 45.0 0.4 mol/Citrate buffer (PH4.5) 25.0 Purified water Remainder Each component was mixed to obtain a base, and per 100g of it,
Interferon was mixed in a proportion of 1×10 ⁶ international units to obtain a gel for external use.

Example 3 Ointment base formulation Ingredients % Carboxymethyl cellulose 0.2 Glycerin 30.0 White petrolatum 20 Stearyl alcohol 22 0.4 mol/Citrate buffer 25 Purified water Residual ingredients were mixed to obtain a base, and per 100 g of the
An ointment was obtained by mixing interferon in a proportion of 1×10 ⁶ international units.

Example 4 Pasta base formulation Ingredients % Carboxymethyl cellulose 2.0 Glycerin 25.0 Setanol 2.8 Glyceryl monostearate 9.3 Tween 80 2.0 Glucuronic acid 1.0 0.4 mol/Citrate buffer 20 Purified water Remainder Mix each component to obtain a base, and 100 g of it Hit,
A paste agent was obtained by mixing interferon in a proportion of 1×10 ⁴ international units.

Example 5 Liquid base formulation Ingredients % Carboxymethyl cellulose 0.1 Glycerin 15 0.4 mol/Citrate buffer (PH4.5) 50 Purified water Remainder Mix each component to obtain a base, and interferon per 100 g of the base is 1×10 ⁵ A solution was obtained by mixing in international unit proportions.

Example 6 Spray formulation Ingredients % Interferon solution obtained in Example 5 50 Freon 114 50 The solution was filled into a 50 ml aluminum container with a Teflon-treated inner surface along with Freon 114 to obtain a spray.

Safety test (1) Primary skin irritation test New Zealand white female rabbit (weight
2.5-3.0Kg) Remove hair from the backs of 6 birds, and place
The gels of Examples 1 and 2 were respectively applied and sealed for 24 hours. The applied area was observed 30 minutes after the gel was removed, but no abnormalities were observed.

(2) Skin sensitization test Female white guinea pigs (body weight 300-350)
g) Example 1 using 23 fish by intradermal injection sensitization method
and 2 gel agents were investigated. As a result, no sensitization was observed other than the effect of human serum albumin, which is contained as a stabilizer in interferon, the raw material. Furthermore, a 24-hour sealed patch test of these gels was conducted to elicit reactions, but no positive cases were observed.

(3) Human patch test A 24-hour closed patch test of the gels of Examples 1 and 2 was conducted on the upper backs of 10 healthy adult males. The patched area was observed 60 minutes after the gel was removed, but no abnormalities were observed.

(4) Human oral mucosal irritation test The gels of Examples 1 and 2 were applied to the oral mucosa of eight healthy adult males, covered with the oral mucosal adhesive bandage, and after 60 minutes the bandage was removed and observed. No abnormalities were observed in either case.

Thus, the external preparation of the present invention has very high topical safety.

Clinical trial Example of efficacy test for lichen planus 1 Subject: 73 years old, female At the first examination, red lesions with extensive edema were observed on both cheek mucosa, with some lace-like white streaks and white coating mixed together. It was done. Both clinically and histologically, the diagnosis was lichen planus. Therefore, vitamins A, B complex, and C were administered and the progress was observed, but there was no change in the disease state. Then, application of the gel of Example 1 was started. 1st day, 3rd day, 7th day after start of application
When applied 4 times, once on day 1 and 12, the red lesions slightly intensified after the fourth application.
It has been well localized from the mucosa around the lesion. Furthermore, on the 22nd day, 36th day, 43rd day, 48th day,
When the gel was continued to be applied on the 57th and 62nd days, the lesion area shrunk after the 62nd day (10th application), and the treatment was completed after 3 applications.
At this time, the oral mucosal lesions had almost completely disappeared. No further recurrence is allowed.

Test Example 2 Subject: 51 years old, female At the time of initial examination, red lesions on the oral mucosa extending from the mandibular buccal gingiva to the posterior molar gingiva, and near the opening of Stensen's canal on the buccal mucosa, including erosion, ulceration, and white coating. and a mixture of white streaks was observed. Both clinically and histologically, the diagnosis was lichen planus. Application of the gel of Example 1 was started while anti-inflammatory treatment was being performed. After the start of application, it was applied 4 times, once on the 1st, 3rd, 6th, and 10th day. From this section, the boundary between the mucosal lesion and the surrounding mucosa has become clearer, but the red lesion has become more intense and hemorrhagic. When the gel was further applied, from around the 7th application (30th day), it was observed that the Witskam's striae tended to disappear and the bleeding was reduced, and the mucosal lesions were localized and reduced in size. A total of 12 gel applications were performed until the 62nd day. A reduction in oral mucosal lesions was observed. Afterwards, the disease condition stabilized, so we performed excision of the remaining oral mucosal lesion and skin grafting. The mucosal lesions were almost completely cured, and no recurrence was observed.

Test Example 3 Subject: 73 years old, male The subject underwent excision of the primary lesion and upper cervical lymph node dissection for treatment of recurrent oral mucosal carcinoma. Subsequently, lichen planus was observed to develop from the gingiva of the left mandibular posterior molar to the buccal mucosa. Therefore, application of the gel of Example 2 was started. After application three times, once on the 1st, 5th, and 10th day, the mucosal lesions increased in redness and swelling for a while, but
After the 7th application on the 25th day, there was a tendency for the erosion to improve, and if the application was continued,
His condition gradually improved. On the 62nd day, the 11th application was performed and the treatment was completed. The lesion has almost completely healed, and no recurrence has been observed.

Example of efficacy test for leukoplakia 1 Subject: 63 years old, female The subject had developed vitiligo and erosion on the dorsum of the left tongue, and despite continued application of steroid ointment, no improvement in symptoms was observed. Vitiligo, ulcer formation, and spontaneous pain were observed from the tongue margins to the tip of the tongue on both sides. The patient was diagnosed with leukoplakia both clinically and pathologically. Therefore, application of the gel of Example 1 was started. Leukoplakia almost disappeared after the fifth application on the 46th day after the start of application, and no recurrence was observed thereafter.

Test Example 2 Subject: 63 years old, male At the first examination, a white lesion was observed on the left buccal mucosa in an area of approximately 2.5 x 2 cm from below the opening of the stainless steel canal to the rear molars, which could not be removed by rubbing. The surface was partially granular and had mucosal erosion and edema. However, no significant ulcer formation was observed.
The patient was diagnosed with leukoplakia both clinically and by pathological diagnosis. Therefore, application of the gel of Example 1 was started. From around the 4th application on the 19th day, the leukoplakia lesions on the buccal mucosa began to disappear, and by the 10th application on the 62nd day, only a white line showing a tendency to keratinization remained, and contact pain The symptoms almost resolved, and the mucosal erosion and red discoloration also disappeared. In this case, treatment was completed after the 10th application, but although white streaks with a tendency to keratinization remained, there were no subjective symptoms and no tendency for recurrence was observed.

Test Example 3 Subject: 70 years old, female A mucosal lesion containing redness of the mucous membrane, erosion, and a white lesion that could not be removed by rubbing was observed in the area extending from the right lower lip to the buccal mucosa and the crestal gingiva of the molar region. The patient was diagnosed with leukoplakia both clinically and by pathological diagnosis. Therefore, application of the gel of Example 2 was started. After the third application on the 27th day, the white moss on the oral mucosa gradually decreased. moreover,
The redness of the mucous membrane initially seemed to increase, but it gradually approached the normal mucosal image, and after the 10th application on the 84th day, the leukoplakia lesions almost disappeared, and the subjective symptoms such as contact disease also disappeared. did.

As shown in the above clinical test examples, the external preparation of the present invention exhibits extremely excellent therapeutic effects on lichen planus and leukoplakia. During each clinical trial, no abnormalities were observed in the subjects' blood cell counts or biochemical values.

Claims (1)

[Scope of Claims] 1. An external preparation for treating dyskeratosis of the skin and mucous membranes, which contains interferon as an active ingredient, and contains a trivalent or higher valent sugar alcohol and an organic acid buffer. 2. The external preparation of item 1 above for the treatment of lichen planus. 3. The external preparation according to item 1 above, which is for the treatment of leukoplakia. 4. The external preparation according to any one of Items 1 to 3 above, containing 10 to 10 ⁸ international units of interferon per dosage unit. 5. The external preparation according to any one of Items 1 to 4 above, which is in the form of an ointment. 6. The external preparation according to any one of Items 1 to 4 above, which is in the form of a pasta preparation. 7. The external preparation according to any one of Items 1 to 4 above, which is in the form of a gel preparation. 8. The external preparation according to any one of Items 5 to 7 above, which contains carboxymethyl cellulose as a binder. 9. The external preparation according to any one of the above items 1 to 4, which is in the form of a spray. 10. The external preparation according to any one of Items 1 to 4 above, which is in the form of a liquid preparation.