WO2008047779A1

WO2008047779A1 - Therapeutic or prophylactic agent for atopic dermatitis

Info

Publication number: WO2008047779A1
Application number: PCT/JP2007/070144
Authority: WO
Inventors: Akira Asari
Original assignee: Glycoscience Laboratories, Inc.
Priority date: 2006-10-17
Filing date: 2007-10-16
Publication date: 2008-04-24
Also published as: JPWO2008047779A1; JP5457675B2

Abstract

[PROBLEMS] To provide a pharmaceutical composition for the prevention and/or treatment of atopic dermatitis. [MEANS FOR SOLVING PROBLEMS] Disclosed is a therapeutic or prophylactic agent for atopic dermatitis, which comprises a low-molecular-weight hyaluronic acid as an active ingredient. The low-molecular-weight hyaluronic acid may be hyaluronic acid di- to hexa-saccharide, and hyaluronic acid tetrasaccharide is preferably used.

Description

Specification

Therapeutic or preventive for atopic dermatitis

Technical field

[0001] The present invention relates to prevention and prevention of atopic dermatitis comprising low molecular weight hyaluronic acid as an active ingredient.

It relates to a pharmaceutical composition for treatment.

Background art

[0002] Atopic dermatitis is an allergic constitution called atopic predisposition, such as allergic asthma, allergic rhinitis, dermatitis, and chronic skin with itching caused by various stimuli. Is a disease. The diagnostic criteria of the Japanese Dermatological Association define skin diseases mainly consisting of eczema with itching and atopic dermatitis that repeat exacerbations and remissions, and many patients have the ability to have atopic predisposition. Not necessary. Therefore, the term “atopic dermatitis” may be used for diagnosis, including contact dermatitis. In general, it exhibits chronic eczema symptoms over a wide area, becomes dry and blows white powder on the surface, and is accompanied by strong itching. Some patients may have red eczema, nodules, etc.

Tissue fluid leaches out from the wet phase, causing severe pain. When it becomes chronic, it becomes rough as if it were goosebumps, and the skin gradually thickens or sometimes has a lump-like wart-like rash.

[0003] Atopic dermatitis develops in early childhood and often remits spontaneously before reaching adulthood.

However, there are an increasing number of cases that carry over to adults, and cases that develop or recur after adulthood. Basically, the goal is to alleviate the symptoms by symptomatic treatment with anti-itch drugs such as steroids and antihistamines. Treatment that suppresses excessive immunity by administration of external or internal steroids, or protects the dried part with a moisturizer such as petrolatum is the mainstream.

[0004] However, steroids that are said to have the highest therapeutic effect! / Are caused by a phenomenon called rebound that rebounds more strongly than the original symptom, and the effect is reduced by long-term continuous use. (Takifiraki 1) may occur. In addition, topical steroids have been used for a long time Then, it is known that skin atrophy, induction of skin infection, and telangiectasia appear, and it is difficult to control steroid dosage and administration period according to symptoms. There has been a demand for the development of therapeutic agents with fewer side effects than steroids.

[0005] In order to prevent drying due to atopic dermatitis, therapeutic agents containing hyaluronic acid as a moisturizing agent have been developed. For example, Patent Document 1 discloses a therapeutic drug for atopic dermatitis containing ε-polylysine as an active ingredient and hyaluronic acid as a humectant. Patent Document 2 describes that a sheet in which hyaluronic acid and hydrolyzed silk are contained in a silk nonwoven fabric is used for the prevention and treatment of atopic dermatitis. Patent Document 3 discloses an external preparation for skin containing borage seed oil and vegetable ceramide extracted from wheat germ as main components and hyaluronic acid as a functional component. Patent Document 4 describes a skin external preparation containing eicosapentaenoic acid or docosahexaenoic acid and hyaluronic acid as a moisturizing agent. However, hyaluronic acid is used only as a moisturizing agent and is not contained as an active ingredient in the treatment of atopic dermatitis. Hyaluronic acid is so-called high molecular weight hyaluronic acid (for example, weight average molecular weight of about 900,000) and does not contain low molecular weight hyaluronic acid, particularly hyaluronic acid tetrasaccharide.

Patent Document 1: Japanese Unexamined Patent Application Publication No. 2005-035914

Patent Document 2: Japanese Patent Laid-Open No. 2003-212715

Patent Document 3: Japanese Patent Laid-Open No. 2002-053428

Patent Document 4: JP 2000-095683 A

Disclosure of the invention

Problems to be solved by the invention

[0006] An object of the present invention is to provide a therapeutic agent with a high therapeutic effect that can be substituted for a steroid agent in the prevention and / or treatment of atopic dermatitis.

Means for solving the problem

[0007] The present inventor has found that low molecular weight hyaluronic acid, particularly hyaluronic acid tetrasaccharide, has an effect of suppressing inflammation due to atopic dermatitis, and that administration can reduce symptoms of atopic dermatitis. I found it. [0008] That is, the present invention provides a therapeutic or prophylactic agent for atopic dermatitis comprising low molecular weight hyaluronic acid as an active ingredient.

[0009] Further, in the present invention, low molecular weight hyaluronic acid is disaccharide (molecular weight about 400), trisaccharide (molecular weight about 400).

600), tetrasaccharide (molecular weight about 800), pentasaccharide (molecular weight about 1000) or hexasaccharide (molecular weight 1200). Further, it is preferably a low molecular weight hyaluronic acid power hyaluronic acid tetrasaccharide (molecular weight of about 800).

The invention's effect

[0010] According to the present invention, it is possible to provide a therapeutic agent having a high therapeutic effect and having no side effects or having no side effects when preventing and / or treating atopic dermatitis.

Brief Description of Drawings

[0011] FIG. 1 shows the results of measuring the thickness of the left auricle of an atopic dermatitis model mouse in Example 1. FIG. 2 is a color photograph of an affected area of an atopic dermatitis model mouse in Example 1.

3] Observe the behavior of atopic dermatitis model mice in Example 1 (times / 15 minutes)

FIG. 4 is a color photograph taken of an affected area of a patient in Example 2.

BEST MODE FOR CARRYING OUT THE INVENTION

[0012] Hyaluronic acid is a macromolecular long-chain polysaccharide composed of a disaccharide repeating unit of D-glucuronic acid and N-acetyl-D-darcosamine, while oligosaccharides are also known. The molecular weight hyaluronic acid is a hyaluronic acid disaccharide, trisaccharide, tetrasaccharide, pentasaccharide, or hexasaccharide (molecular weight of about 400 to about 1200), and particularly, hyanolonic acid tetrasaccharide.

[0013] As the low molecular weight hyaluronic acid contained in the drug of the present invention, basically, the 1-position of / 3-D-glucuronic acid and the 3-position of β D-Ν-acetyl-darcosamine 1 were combined. As long as it contains at least one disaccharide unit and has more than two sugars and is basically composed of β-D-glucuronic acid and β D-N-acetyleno-darcosamine, These derivatives may be sugars in which those elements are bonded to multiple bonds, and these derivatives, For example, those having a hydrolyzable protecting group such as an acyl group can be used. Examples of unsaturated sugars that may be unsaturated sugars include non-reducing terminal sugars, usually those in which the 4th and 5th carbon positions of glucuronic acid are unsaturated.

[0014] Specific examples of the low molecular weight hyaluronic acid used in the present invention include those extracted from natural products such as animals, those obtained by culturing microorganisms, and chemically or enzymatically synthesized. Any of these can be used. For example, it can be obtained by a known tissue extraction method and purification method, such as a living tissue force such as a chicken crown, trousers, skin, and joint fluid. It can also be produced by a fermentation method using bacteria belonging to the genus Streptococcus.

[0015] In the present invention, low molecular weight hyaluronic acid such as a disaccharide consisting of one disaccharide unit and a derivative thereof can be used. Preferably, low molecular weight hyaluronic acid of about 2 to 20 sugars can be mentioned, more preferably low molecular weight hyaluronic acid of 2 to 6 sugars (molecular weight of about 400 to about 1200), most preferably hyaluronic acid tetrasaccharide. It is.

[0016] Specifically, low molecular weight hyaluronic acid is produced by known methods such as an enzymatic decomposition method, an alkaline decomposition method, a heat treatment method, and an ultrasonic treatment method (Biochem., 33 (1994) p6503-6507). The acid is preferably produced by a method of reducing the molecular weight, a method of chemically or enzymatically synthesizing (Glycoconjugate J., (1993) p435-439, WO93 / 20827), and the like. For example, enzymatic degradation methods include hyaluronic acid-degrading enzymes (hyaluronidase (derived from testicles), hyaluronidase (derived from Treptomyces), hyaluronidase SD, etc.), chondroitinase AC, chondroitinase ACII, chondroitinase ACIII, chondroitinase ABC, etc. A method for producing hyaluronan oligosaccharides by causing an enzyme that degrades hyaluronan to act on hyaluronan (Seminar of Biochemical Experiments "Carbohydrate II Proteodarican and Glycosaminodarlican" p244-248, 199 1 year, Tokyo Chemical Dojin For example).

[0017] In addition, as an alkali decomposition method, for example, a base such as about 1N sodium hydroxide is added to a hyaluronic acid solution, heated for several hours to lower the molecular weight, and then an acid such as hydrochloric acid is added. And a method of neutralizing to obtain low molecular weight hyaluronic acid. Hyaluronic acid used in the present invention includes a pharmacologically acceptable salt form, and a pharmaceutically acceptable salt thereof can be used as necessary in the preparation. For example, alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as calcium salt and magnesium salt, tri (n A) amine salts such as amine salts, triethylenoamine salts, pyridine salts and amino acid salts.

[0018] The agent of the present invention can be used without any particular limitation, such as hyaluronic acid having a specific molecular weight alone or a mixture of hyaluronic acids having different molecular weights. The therapeutic agent for atopic dermatitis of the present invention comprises hyaluronic acid as an active ingredient, and by administering an effective amount thereof to mammals including humans, symptoms of atopic dermatitis without adversely affecting the living body. Can be improved or prevented.

[0019] The drug of the present invention can be applied to the affected area and the surrounding skin with low molecular weight hyaluronic acid or a salt thereof as it is or together with a carrier, excipient, or other additives as necessary. Alternatively, it can be formulated into an arbitrary dosage form and administered intra-tissue (injection), orally or menstrually.

[0020] Examples of the dosage form of the drug to be applied to the skin include ointments, lotions, and creams. The base for these external preparations for skin is not particularly limited as long as it is a base that can be melted, blended or dispersed uniformly in an external preparation. For example, petroleum jelly, castor oil, silicon, squalene, sodium talinoleate, behenyl alcohol, glycerol monostearate, stearyl alcohol, ethanol, batyl alcohol, phenoxyethanol, 1,3-butylene glycol, isopropyl myristate, stearin Examples thereof include, but are not limited to, acid, lecithin, purified water and the like.

[0021] The agent of the present invention may be added with an antioxidant, an antiseptic, a wetting agent, a thickening agent, a buffering agent, an adsorbent, a solvent, an emulsifier, a stabilizer, a surfactant, and the like, if necessary. Agent can be added

Yes

[0022] An appropriate amount of an ointment containing about 0.05 to 5% of low molecular weight hyaluronic acid can be applied to the affected area and its surroundings several times a day. For example, hyaluronic acid tetrasaccharide can be applied to the affected area at a concentration of 1%. When used for the prevention of atopic dermatitis, apply in the same way to areas where atopic dermatitis is likely to develop.

[0023] The drug of the present invention can be used as a therapeutic agent for suppressing the progression of symptoms of atopic dermatitis or alleviating the symptoms by applying to the affected area. Applied to the skin or who has a predisposition for atopy but has not developed symptoms, or a patient with atopic dermatitis It can be applied to people who are highly likely to develop atopic dermatitis, such as those in their families, and can be used as a preventive agent to prevent recurrence of atopic dermatitis.

Example

Hereinafter, the present invention will be described more specifically with reference to examples. However, the technical scope of the present invention is not limited to these examples.

[Example 1: Confirmation of the therapeutic effect of HA4 using atopic dermatitis model mice]

Inflammation of the left auricle was produced by sensitization and induction with PiCl in NC mice, and the effect of the HA 4 formulation was examined. As a result, it was observed that inflammation scores such as skin dryness, crust formation, bleeding, hair loss, wrinkling behavior, and auricular thickness associated with inflammation decreased.

[0025] <Creation of dermatitis model mouse>

Twelve male 9-week-old NC / Nga Sic mice were obtained from Japan SLC Co., Ltd., and the appearance was examined at the time of acceptance to confirm that all cases were normal. Breeding is performed in a plastic cage (14.5 X 26. 0 X 12.) in a breeding room set at a temperature of 22 ± 2 ° C, a relative humidity of 55 ± 15%, a ventilation rate of 12 times / hour, and a light / dark time of 12 hours. 5 cm) were raised one by one. The feed was a solid feed Roden Lab Diet EQ (Nihon SLC Inc., LotNo. AUG24062), and the drinking water was freely taken daily by public tap water. After acclimatization for 9 days, 10 animals with good weight gain, general condition and manipulation site during the acclimatization period were selected and randomly divided into 2 groups of 5 animals.

[0026] For all cases, 0.05 mL per mouse was applied to the abdomen of a 5% PiCl / ethanol 4: acetone 1 solution using a syringe for disposable wurzbergrin on the abdomen of a mouse previously depigmented with a hair clipper, 0 · 02 mL per limb was applied to each limb. In addition, using a syringe for Disposer Brubber Clin, from 4 days after sensitization, l% PiCl olive oil solution was applied to the left auricle of the mouse at 0.02 mL, 3 times a week, and inflammation of the left auricle. Make sure to induce the application and create a skin inflammation model.

The test substance, HA4, is specifically prepared by the method of Tawada et al. (Tawada A, Masa T, Oonuki γ, Watanabe A, Matsuzaki Y, Asan A. Large-scale preparation, purirication, and characterization of hyaluronan oligosaccharides from 4- mers to 52-mers. Glycobiolog y. 2002; 12 (7): 421-6 ·). As the test substance IV-4 formulation, a formulation formulated as shown in Table 1 was used. As a control substance, a base-only coating agent not containing ΗΑ4 was used. Of the ingredients contained in this drug, except for 以外 4, it is a general base material and is known to be effective for skin diseases such as atopic dermatitis.

[table 1]

[0029] <Test substance administration>

In Group 1, the test substance listed in Table 1 HA4 formulation (manufactured by Glucose Research Institute, Inc .: LotNo.060607, HA formulation amount lmg / ml) 1 hour before the third induction Once a day, 0.02 mL was applied to the left auricle using a disposable Brucellin syringe. The HA4 formulation was administered for 5 weeks after the 3rd induction. The remaining 1 group was administered with the control substances listed in Table 1 (manufactured by Glucose Science Laboratory: Lot No. 060607) as a negative control (Table 2).

[0030] [Table 2] Group Administration category Number of animals (units) Test substance group Sensitization / induction with P i C 1 and application of HA 4 5

Control group P i C 1 cropping ■ Induction and base only (control) 5

[0031] <Evaluation>

The general condition of each mouse was observed at least once a day, and the body weight was measured at least once a week on the sensitization date, the test end date, and other days. Furthermore, the Student t t test was performed on the body weight and the scores of the following evaluation items.

[0032] (1) Auricular thickness measurement

Before the sensitization, before each induction application, and before each administration once a week from the start of administration of the test substance, the thickness of the left auricle of the mouse was measured using a mic mouth meter (manufactured by Mitutoyo). For reference, I also measured the right auricle! /.

(2) Observation of skin symptoms

Immediately before each induction application and once a week from the start of test substance administration,

The following items were observed and evaluated.

Observation items: Redness, edema, hair loss, desiccation, crust formation, bleeding, ulcer, tissue defect, etc. Evaluation score: asymptomatic (score 0), mild (score 1), moderate (score 2), severe (score 3)

(3) Observation of drought behavior

From 1 hour after each induction application and once a week from the start of test substance administration, from 1 hour after each administration, the wrinkling behavior of the applied part was observed for about 15 minutes, and the number of pulling was counted. Photographs of the mouse pinna of all cases were taken on the test end date.

[0033] <Test results and discussion>

There was no significant difference in body weight in the test substance group compared to the control group, and there was no abnormality except that both the control group and the test substance group showed atopic skin inflammation. [0034] 1. Measurement of pinna thickness (Figure 1)

The left auricle rose after sensitization in both the control group and the test substance group, and increased 2 weeks after the start of induction, and after 2 weeks, both groups reached about 2.1 times the thickness at the time of sensitization. She had symptoms of atopic dermatitis.

On the day following the administration of HA4 over 5 weeks (36th day of administration), the control group had 1.48 times the same and the test substance group had 1.39 times, and the thickness of the left auricle of the test substance group was higher than that of the control group. Significantly decreased. The left auricle thickness in the test substance group showed a lower value compared to the control group at each measurement point on days 9, 16, 23, and 30 after the start of administration, and there was no significant difference. . The thickness of the right auricle measured as a reference was 5% lower than that of the control group on the day after the 35th day of administration, and the test substance group showed a low value.

[0035] 2. Observation of skin symptoms

(Redness)

In the control group, 2/5 cases were observed in the left auricular skin at the second induction, all cases at the third induction, and 2/5 cases from the control group on the 9th day of administration. Mild redness was observed.

In the test substance group, mild redness was observed in 3/5 cases during the second induction and in all cases during the third induction. On the 9th day of administration, 1/5 patients in the test substance group had mild redness, and on the 23rd day, 1/5 patients in the test substance group had mild redness.

[0036] (Edema)

3rd induction • Mild edema was observed in all groups on day 1, day 9, and day 16 of administration in both groups.

(Dry)

In the control group, mild dryness was observed in all cases at the 2nd and 3rd induction, 2/5 cases on the 9th day of administration, 1/5 cases on the 16th day and 23rd day, and mild dryness. Admitted.

In the test substance group, 4/5 cases at the 2nd induction, all cases at the 3rd induction, 1/5 cases on the 9th day of administration, mild dryness was observed.

[0037] (Crust formation 'bleeding')

Mild in all controls in the 2nd induction, moderate in 1/5 in the 3rd induction, mild in 3/5, 4/5 on the 9th day of administration, 1 / day on the 16th day of administration 5 cases, 2/5 cases light on day 23 Degree of crust formation · Bleeding was observed.

Mild scab formation / bleeding was observed in all test substance groups at the second induction, 3/5 at the third induction, 2/5 at 9th day of administration, and 1/5 at 16th day of administration It was.

None of the cases had skin ulcer tissue defects.

[0038] (Hair removal)

Hair loss was observed in all cases in both groups at the third induction and on the ninth day of administration. On the 16th day of administration, 4/5 cases were observed in the control group and 2/5 cases in the test substance group, and the test substance group showed a significantly lower value at the 5% level compared to the control group. Fig. 2 shows photographs of the left pinna of typical individuals in the control group and the test substance group on the 30th day after administration.

[0039] As described above, there are cases in which the test substance group shows a lower value than the control group in each item of redness, edema, hair loss, dryness, and crust formation. It showed a low value. In all cases, there were no ulcers or tissue defects in the inflamed areas.

[0040] 3. Observation of drought behavior

Figure 3 shows a graph summarizing the observation results of drought behavior for the control group and the test substance group. In both groups, the number of sputum at the first induction was high at the second induction, and only the test substance group was higher at the third induction.

From day 9 to day 35 of administration, the number of sputum gradually decreased in both groups, except for the high value in the control group on day 30 of administration. The test substance group on the 23rd, 30th, and 35th day after administration (36th day) showed a lower value than the control group, but no significant difference was observed.

[0041] As described above, administration of a test substance to inflammation produced by induction of PiCl sensitization in NC mice reduced the inflammation scores such as hair loss, wrinkling behavior and auricle thickness. These results indicate that administration of HA4 alleviates symptoms in atopic dermatitis model mice, and strongly suggests that HA4 is effective in the treatment of atopic dermatitis.

[0042] In addition, the thickness of the right auricle, which is the opposite side to which the test substance was applied, was significantly increased compared to the control group. This suggests that the entire body was moved to the other side. This strongly suggests a successful treatment for atopic dermatitis with systemic symptoms. [Example 2: Confirmation of therapeutic effect of HA4 in patients with atopic dermatitis] As the agent of the present invention, the HA4 combination coating agent shown in Table 1 was used.

[0044] A 16-year-old female (height: 169cm, weight: 50kg) suffering from atopic dermatitis was volunteered and the cream was applied. The patient was diagnosed with atopic dermatitis at an early age and was not completely cured. The patient presents with mild, chronic lesions such as infiltrative erythema, prurigo, crust, rupture, papules, and wrinkles. The patient was applied the above cream on the inside of the elbow for a week after bathing in the morning and evening. One week after administration, chronic lesions and wrinkles disappeared (Figure 4). Other than hyaluronic acid tetrasaccharide, it is a general base material and is not known to be effective in treating atopic dermatitis. Therefore, this effect is considered to be the action of hyaluronic acid tetrasaccharide, suggesting that hyaluronic acid tetrasaccharide is effective in the treatment of atopic dermatitis.

Claims

The scope of the claims

[1] A therapeutic or prophylactic agent for atopic dermatitis comprising low molecular weight hyaluronic acid as an active ingredient.

[2] The therapeutic or prophylactic agent according to claim 1, wherein the low molecular weight hyaluronic acid is a disaccharide to hexasaccharide low molecular weight hyaluronic acid.

[3] The treatment or prevention according to claim 1, which is a low molecular weight hyaluronic acid power hyaluronic acid tetrasaccharide.

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