JP2006306813A - Mast cell increase inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new composition effective for preventing or treating diseases related to an increase in mast cell count. <P>SOLUTION: The composition effective for preventing or treating diseases related to an increase in its cell count contains dihomo-γ-linolenic acid (DGLA). <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention is a food having a prophylactic or therapeutic effect against various pathological conditions in which an increase in the number of mast cells is observed in the lesion, such as atopic dermatitis, bronchial asthma, hay fever, allergic rhinitis, and allergic conjunctivitis The present invention relates to an ingredient or a food composition or a pharmaceutical composition containing the ingredient, and particularly relates to an inhibitor for increasing the number of mast cells.

  In recent years, the number of allergic diseases has increased dramatically due to changes in the environment and diet, genetic factors, and the like. The pathology of allergic diseases is largely classified from type I to type IV based on the pathogenesis mechanism and related functional molecules, and mast cells are deeply related to one of the immediate allergies called type I allergy. It is said that. In the case of type I allergy, when a living body is exposed to a certain antigen, antigen-specific IgE antibodies are produced by B cells via antigen-presenting cells and helper T cells.

  The IgE antibody then binds to the surface of the mast cells, and becomes a state to watch for the invasion of the antigen again. The mast cells in this state capture the antigen that has invaded again on the membrane surface and degranulate. Causes various chemical mediators such as histamine and leukotrienes. Subsequently, these chemical messengers bind to the receptor, causing undesired physiological phenomena such as edema, redness, itching, airway resistance and increased mucus secretion, which are so-called “allergic reactions”.

  Against this background, the method of inhibiting mast cell function, specifically the method of inhibiting mast cell degranulation and inhibiting the release of chemical mediators, the method of inhibiting the synthesis of chemical mediators, or released Attempts have been made to reduce allergic reactions, particularly by inhibiting the action of chemical mediators produced by mast cells, such as methods for inhibiting the binding between chemical mediators and receptors. Currently, it has been found that many of the drugs used for the treatment of itching of atopic dermatitis, hay fever, allergic conjunctivitis, etc. are based on one of the mechanisms shown above (Internet HP "Rheumatology and Allergy Information Center", Hasegawa Yuki, 2005.04.04, Allergic Inflammatory Diseases-A New Approach-, Motohiro Kurosawa, 1994).

  However, there is an example which reports that there is a change not only in the function of the chemical transmitter produced by mast cells as described above but also in the number of mast cells confirmed in the lesion. For example, it has been found that the differentiation and proliferation of mast cells in the nasal mucosa of allergic rhinitis patients tend to be higher than that of non-allergic ones (Journal of the Otolaryngology Society of Japan, Naofumi Yoshida, 2001, 104, p.504-). In addition, it has been shown that the number of mast cells localized in the bronchial smooth muscle bundle is significantly higher in asthmatic patients than in healthy individuals (Current Opinion in Allergy and Clinical Immunology, Peter B, 2003). , 3, p.45-).

  Similar reports have been made on skin diseases. For example, an increase in mast cells has been reported in basal cell carcinoma, psoriasis vulgaris, atopic dermatitis lesions, etc. (Allergy, Noboru Yamamoto, 2000, 49, p. 455 ~). In other words, as a means of suppressing or reducing diseases deeply related to the increase in the number of mast cells, not only controls the amount and function of chemical mediators produced by inflammatory cells, particularly mast cells, but also abnormalities of mast cells themselves. It is also necessary to suppress proliferation and maintain the number of cells at a normal level, and it is assumed that more effective and multifaceted disease prevention and treatment can be realized by controlling this process.

  For example, steroid external preparations and immunosuppressive external preparations that have been clinically useful for allergic dermatitis, etc. are used in NC / Nga mice, which are model animals of the same disease, for dermatitis associated with the onset of dermatitis. In addition to showing a significant improvement in symptom score and a significant suppression of blood IgE elevation, it has also been found to significantly reduce the number of mast cells that increase at the dermatitis lesion (Japanese Journal of Pharmacology). , Hiroi J, 1998, 76, p. 175-), which strongly suggests that the therapeutic effect can be correlated with the number of mast cells, and the importance of controlling the number of mast cells to an appropriate level. Is to support.

Thus, while these drugs have been shown to be useful in the prevention and treatment of various mast cell-related diseases including allergic diseases, it is also true that they have problems in terms of safety and usage It is. If it is a topical steroid, side effects such as flushing during use, skin atrophy, and rebound phenomenon that exacerbates dermatitis due to discontinuation of administration, and if it is an immunosuppressive topical, the onset of skin tumors may be accelerated. It has been reported that the medicinal effect is greatly influenced by the state of the application site and the barrier function of the skin (Journal of Japanese Dermatological Association, Masataka Furue, 2004, 114, p.135-).
Against this background, a material that is medically useful, can be used safely for anyone, and effectively suppresses the increase in the number of mast cells has been demanded.

Hasegawa Yuki, 2005.04.04, Allergic Inflammatory Diseases-A New Approach-, Kurosawa Motohiro, 199 Bulletin of the Japan Otolaryngology Society, Takashi Yoshida, 2001, 104, p. 504- Current Opinion in Allergy and Clinical Immunology, Peter B, 2003, 3, p.45〜 Japanese Journal of Pharmacology, Hiroi J, 1998, 76, p.175〜 Journal of Japanese Dermatological Association, Masutaka Furue, 2004, 114, p.135〜 British Medical Journal, Kernoff PBA, 1977, 2, p. 1441- Lipids, Taki H, 1993, 28, p.873〜 Lipids, Cedric H, 1984, 19, p.699〜

Immunology, Maaike MBWD, 2003, 110, p.348〜 The Journal of Immunology, Deniela S, 1989, 143, p.1303〜 Archives of Dermatological Research, Iverson L, 1992, 284, p. 222〜 Prostaglandine Leukotrienes and Essential Fatty Acids, Zurier RB, 1999, 60, p.371〜 Prostaglandine Leukotrienes and Essential Fatty Acids, Gueck T, 2003, 68, p.317〜 Veterinary Dermatology, Gueck T, 2004, 15, p.309〜

  An object of the present invention is to provide a food or pharmaceutical composition for safe or more effective prevention or treatment for various diseases closely related to an increase in the number of mast cells.

As a result of intensive studies to solve the above-mentioned problems, the present inventor found for the first time that dihomo-γ-linolenic acid (DGLA) is extremely useful for suppressing the increase in the number of mast cells, and completed the present invention.
Considering medically and nutritionally useful food ingredients, dihomo-γ-linolenic acid (DGLA), an n-6 polyunsaturated fatty acid (PUFA) as shown in FIG. It can be cited as one of the candidate materials. DGLA is contained in various foods such as meat, eggs, seafood, etc., and these have been ingested regularly by human beings for many years and it goes without saying that they are extremely safe materials. . However, it has been found that the total content is much lower than that of the same n-6 PUFA arachidonic acid and n-3 PUFA EPA and DHA. As described in Patent No. 335481, the present inventors have invented a method for fermentative production of DGLA oils and fats by using strains, so that it is possible to prepare a large amount of triglyceride SUNTGD in which about 40% of the constituent fatty acids are composed of DGLA. did.

  The physiological function of this component has been partially confirmed in vivo or in vitro. In vivo, platelet aggregation is inhibited (British Medical Journal, Kernoff PBA, 1977, 2, p. 1441), delayed Type of footpad edema (Lipids, Taki H, 1993, 28, p.873 ~), blood pressure rise suppression (Lipids, Cedric H, 1984, 19, p.699 ~), etc. 2,10, Inhibition of cytokine production such as tumor necrosis factor (TNF-α) (Immunology, Maaike MBWD, 2003, 110, p.348 ~, The Journal of Immunology, Deniela S, 1989, 143, p.1303 ~) , Leukotriene production inhibitory action (Archives of Dermatological Research, Iverson L, 1992, 284, p. 222 ~), T cell proliferation inhibitory action (Prostaglandine Leukotrienes and Essential Fatty Acids, Zurier RB, 1999, 60, p.371 ~), etc. However, there is no direct proof regarding the effect on mast cells.

  On the other hand, linoleic acid (LA) and γ-linolenic acid (GLA) among the same n-6 PUFAs have been studied for their effect on mast cells, and the former is derived from established mast cell types. Has been reported (Prostaglandine Leukotrienes and Essential Fatty Acids, Gueck T, 2003, 68, p. 317-), and the latter suppresses histamine release in the same test system. (Veterinary Dermatology, Gueck T, 2004, 15, p.309-). However, in any of these studies, the effect on the release of chemical mediators from mast cells was examined, and it is not known at all what effect both have on the number of mast cells.

As described above, it was not known at all whether DGLA had any effect on the number of mast cells.
Therefore, the present invention provides a composition comprising DGLA and having a preventive or therapeutic effect on various diseases closely related to an increase in the number of mast cells.
This composition is, for example, a food composition or a pharmaceutical composition.
The various diseases include skin diseases such as atopic dermatitis, psoriasis, psoriasis, basal cell carcinoma, squamous cell carcinoma, bronchial asthma, hay fever, allergic rhinitis, allergic conjunctivitis and the like.

DGLA exists, for example, in the form of glycerides, phospholipids, glycolipids, alkyl esters, or free fatty acids. The glyceride is, for example, triglyceride, diglyceride, or monoglyceride. Preferably, the glyceride is triglyceride and / or diglyceride.
The composition is, for example, in the form of a pill, tablet or capsule.
The composition of the present invention can also take the form of foods and drinks comprising DGLA and displaying that it has a preventive or therapeutic effect on various diseases closely related to increased mast cell count. For example, it can be a food or drink with a label indicating that it has a prophylactic or therapeutic effect on various diseases closely related to an increase in the number of mast cells containing DGLA.

  Oral intake of DGLA can suppress the increase in the number of mast cells extremely safely and effectively, and therefore DGLA can be used in various diseases that are closely related to the increase in the number of mast cells, such as skin diseases, asthma and rhinitis. It is highly useful.

The present invention is described in detail below.
Food compositions include dietary supplements, and (pharmaceutical) formulations and preparations such as tablets, pills and capsules. In addition, solid or liquid foodstuffs such as dairy products (margarine, butter, milk, yogurt), bread, cakes; drings such as beverages (tea, coffee, cocoa, chocolate drinks), fruit juices, soft drinks (eg carbonated drinks) ); Confectionery; oily food (snack, salad dressing, mayonnaise), soup, sauce, carbohydrate rich food (rice, noodles, pasta), fish food, baby food (eg infant formula, as liquid or powder), Examples include pet food and food that can be cooked or cooked in a microwave.

DGLA can be derived from any suitable source. However, there are almost no natural fat sources known to have a high DGLA content, and it is possible to extract them from cow liver, pig kidney, egg yolk, etc., in very small amounts. In recent years, microbial fermentation technology has advanced and may be derived from microorganisms such as fungi, bacteria or yeast.
Suitable fungi are those belonging to the order of the Mucorales, for example Mortierella, Pythium or Entomophyhora. A preferred source of DGLA is from Mortierella. More preferably, it is derived from Mortierella alpina. As shown in Japanese Patent No. 334581, the inventors can prepare triglycerides in which about 40% of the constituent fatty acids are composed of DGLA by microbial fermentation using Mortierella.

In addition to DGLA, one or more additional PUFAs may be supplied. In addition to DGLA, this may be another n-6 PUFA (eg, linoleic acid (LA), γ-linolenic acid (GLA), arachidonic acid (AA), etc.), or n-3 PUFA (eg, EPA, DHA).
The physiologically acceptable functional derivatives of this acid that can be converted to DGLA for use in the present invention include DGLA-containing triglycerides, diglycerides, monoglycerides, or as phospholipids, glycolipids, and free fatty acids and fatty acid esters. (Eg methyl or ethyl ester), may be in the form of a sterol ester.

  Preferably the PUFA is present in the oil. This can be a pure oil, a processed oil (eg a chemically and / or enzymatically treated oil) or a concentrated oil. These oils may contain 10-100% PUFA, but the desired PUFA, eg, DGLA content is 5% or more, preferably 10% or more, more preferably 25% or more of the oil if the oil is derived from a microorganism. If it is good. The oil may contain one or more PUFAs within these percentage concentration ranges. The oil may be a single oil derived from a single cell or microorganism, or it may be a blended or mixed oil of two or more oils derived from other sources. This oil contains, for example, one or more additives such as antioxidants (eg tocopherol, vitamin E, tocotrienol, ascorbic acid derivatives, palmitate or ester, astaxanthin), sesamin, CoQ10, etc. May be.

  The present invention improves the amount of PUFA for healthy individuals who have eaten well and healthy, i.e., individuals with normal PUFA levels in the living body, to prevent disease and maintain and maintain health. Used for purposes.

  However, individuals with low PUFA intake due to inadequate or inappropriate eating habits, etc., or individuals with low PUFA in vivo despite having an appropriate diet, such as n- It can also be used to prevent, prevent, ameliorate, or treat diseases or conditions associated with abnormal or low levels of 3 or n-6 PUFAs. Therefore, the present invention also provides use for subjects with low DGLA levels, such as subjects who cannot convert LA to GLA and DGLA, cannot convert GLA to DGLA, and / or cannot convert effectively. Therefore, suitable patients may have dysfunctional, insufficient or deficient Δ6 desaturase and / or carbon chain extender enzymes.

  In particular, the present invention provides use for subjects with various diseases in which an increase in the number of mast cells is observed. Diseases with increased mast cell count include skin diseases such as atopic dermatitis, eczema, psoriasis, basal cell carcinoma, squamous cell carcinoma, bronchial asthma, hay fever, allergic rhinitis, allergic conjunctivitis, etc. Refers to disease.

Next, the present invention will be described more specifically with reference to examples.
Example 1.
In order to investigate whether or not DGLA intake has any effect on the increase in the number of mast cells, we used triglyceride SUNTGD with DGLA as the main constituent fatty acid prepared by the method shown in Patent No. 334581, which is a DGLA oil. Thus, its usefulness in experimental animals was investigated. In this study, NC / Nga mice were used as a model animal for diseases closely related to an increase in the number of mast cells. This model animal is currently recognized as one of the most useful atopic dermatitis models, and the steroid external preparations and immunosuppressive external preparations that are actually used in the clinical field of atopic treatment are the model animals as described above. Since it has been proved to be effective in the field, it is also widely used for screening for drugs for treating atopic dermatitis.

  This animal developed dermatitis spontaneously around 8 weeks after birth in a conventional rearing environment, and its inflammation became hated and chronic over time, and both macroscopically and histopathologically It is known to exhibit atopic-like skin inflammation. In addition, elevated serum IgE associated with the onset of dermatitis, differentiation / proliferation of mast cells in the lesion, and marked infiltration of immunocompetent cells such as eosinophils and T cells are also characterized by this disease state.

  In this study, male / female NC / Nga mice were prepared, and two groups were established with 7 mice per group in a conventional rearing environment. The following two types of food shown in Table I were prepared and continued to be fed from the 5th week after weaning until the 12th week when the test was terminated by free consumption. The groups were a control diet group and a DGLA diet group. To the latter, triglyceride SUNTGD containing DGLA as the main constituent fatty acid was added, and about 1.0% of the diet was set to be DGLA (calculated as the amount of free fatty acid). Since the average body weight of the mice was 20 g and the average daily food intake was about 2 g, the DGLA intake was calculated to be about 1000 mg / kg per day when the DGLA intake in this experiment was calculated. In both groups, the total amount of fatty acids in the diet was set at 5%. Evaluation items were macroscopic score of skin inflammation under the blind, number of curettages, and blood IgE.

  The animals were dissected at the end of the experiment at 12 weeks of age, and the skin on the back of the neck, which is the dermatitis lesion, was collected, fixed with neutral buffered formalin solution, embedded in paraffin, and sliced sections were prepared. Then, mast cells were identified by staining with toluidine blue. From these tissue specimens, 2 specimens out of 7 specimens in each group were selected, and the total number of mast cells and the degree of the number of these cells were evaluated relative to a total of 4 specimens. As a specimen selection method, a method was adopted in which two specimens showing values close to the average value of the gross score of skin inflammation in each group were selected. Regarding the count of mast cells, count the number of mast cells confirmed in the 10x40 field of view at the time of microscopic examination, repeat this work 5 times for the same sample, changing the field of view, and average the sample value Expressed as the number of mast cells.

Regarding the relative evaluation of the number of mast cells, the number of mast cells in each specimen was relatively determined. The relative evaluation criteria are
++: Several sites with particularly large mast cells were confirmed, and the number of cells was also large overall.
+: Some sites with a large number of mast cells are confirmed, but the number of cells is slightly higher overall.
+-: The number of mast cells is almost normal.
It was. In addition, regarding the count of the number of mast cells and the relative evaluation of the number of the present cells, it was performed under the blind so that it was not known which sample was evaluated.

  As a result of the examination, as shown in Table 3, the number of mast cells increased in the skin lesion as compared to the control diet group in the DGLA intake group without showing any abnormalities in weight transition (Table 2) or general findings. Showed a tendency to suppress. At this time, the gross score of skin inflammation was reduced (control diet group: 9.1 ± 1.0, DGLA diet group: 3.0 ± 0.5), and curettage was suppressed (control diet group: 51.7 ± 9.1 times, DGLA diet group: 35.0 In addition, suppression of plasma IgE production (control diet group: 64.2 ± 39.8 μg / mL, DGLA diet group: 29.8 ± 21.0 μg / mL) was also confirmed. These results suggest that DGLA intake may be very useful for various diseases closely related to increased mast cell count, such as atopic dermatitis, bronchial asthma, and allergic rhinitis.

FIG. 1 shows the metabolic pathways of n-6 and n-3 highly unsaturated fatty acids.

Claims (13)

  A composition comprising dihomo-γ-linolenic acid (DGLA) and effective for preventing or treating a disease associated with an increase in the number of mast cells.   A composition for suppressing the number of mast cells, comprising dihomo-γ-linolenic acid (DGLA).   The composition according to claim 1 or 2, wherein the composition is a food or a pharmaceutical composition.   The composition according to claim 3, wherein the food is a dietary supplement.   The composition according to claim 3, wherein the pharmaceutical composition is an orally administered agent.   The composition according to any one of claims 1 to 5, wherein the composition is a liquid, a pill, a tablet, or a capsule.   The composition according to any one of claims 1 and 3 to 6, wherein the disease associated with an increase in the number of mast cells is a skin disease, a respiratory disease or other allergic disease.   The composition according to claim 7, wherein the skin disease is atopic dermatitis, eczema, psoriasis, basement membrane cell carcinoma, or bite cell carcinoma.   The composition according to claim 7, wherein the respiratory disease is bronchial asthma.   The composition according to claim 7, wherein the allergic disease is hay fever, allergic rhinitis or allergic conjunctivitis.   The composition according to any one of claims 1 to 10, wherein the dihomo-γ-linolenic acid (DGLA) is in the form of glyceride, phospholipid, glycolipid, alkyl ester or free fatty acid or a salt thereof.   12. The composition according to claim 11, wherein the glyceride is triglyceride, diglyceride or monoglyceride.   The composition according to claim 1, wherein the dihomo-γ-linolenic acid (DGLA) is derived from a microorganism.

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