MCP-1 production inhibitor, food for inhibiting MCP-1 production, and oral preparation for inhibiting MCP-1 production

Abstract

To provide a floating particulate matter-induced inflammation suppressant and a food for suppressing floating particulate matter-induced inflammation that are effective at suppressing inflammation that is induced by floating particulate matter such as PM 2.5, yellow dust, and house dust.SOLUTION: The floating particulate matter-induced inflammation suppressant comprises naringenin and / or narirutin as an active ingredient thereof. This floating particulate matter-induced inflammation suppressant also comprises a component of the pericarp of the jabara as an active ingredient thereof. The food for suppressing floating particulate matter-induced inflammation comprises naringenin and / or narirutin as an active ingredient thereof. The food for suppressing floating particulate matter-induced inflammation also comprises a component of the pericarp of the jabara as an active ingredient thereof.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to an MCP-1 Production inhibitor, MCP-1 production food for inhibition, and, MCP-1 production and an oral agent for inhibition.

Background Art

[0002] In order to alleviate allergic symptoms, it is necessary to pay attention not only to allergens but also to allergy exacerbating factors. Humans inhale 15 to 20 m 3 of air per day, and it is suggested that inhalation of polluted air has a great impact on health. In the atmosphere, in addition to allergens such as pollen and mold spores, there are mainly substances such as house dust mainly composed of remains of microorganisms such as mites and mold, PM2.5, yellow sand, tobacco smoke, and chemical substances derived from building materials and fragrances. It is feared that these are involved in the exacerbation of allergic symptoms and the exacerbation of respiratory diseases.

[0003] In recent years, with the rapid industrialization of neighboring countries, air pollution in the local area has been worsening. Due to the westerly wind, the air pollutants are carried over, so the problem of air pollutants typified by PM2.5 has also become apparent in our country. In addition, desertification is progressing due to climate change and overgrazing of livestock in arid regions. Therefore, in the western part of the desert region in East Asia, yellow sand blows in on the westerly wind. In the Chinese and Korean mainlands, the arrival of yellow sand at a level that seriously affects daily life, such as extremely poor visibility, has been observed every year. In our country as well, the scattering of yellow sand has become a problem like pollen and PM2.5, and predictions of scattering and warnings are given in weather forecasts. These are considered to be exacerbating factors for allergic patients, and it is also important to deal with yellow sand-induced inflammation.

[0004] On the other hand, apart from outdoor-derived air pollutants, one must not overlook the suspended substances derived from microbial contamination in the indoor environment when considering air pollutants. Microbial contamination represented by mites and mold is often collectively referred to as house dust, which has the characteristic that the individual occupants must take measures. Outdoor-derived air pollutants such as pollen, PM2.5, and yellow sand do not grow indoors, but since mites and mold are living organisms, if left unattended, they may continue to increase within the dwelling. In particular, due to the emergence of highly insulated and airtight houses in recent years, mites can now live throughout the year and are deeply related to human life. Mites feed on dandruff, dirt, and mold, and their habitats include bedding such as futons and carpets. That is to say, "the living space of humans" can be said to be the same as "the habitat of mites". Since the largest source of food for mites is humans, it is said that if left unattended, mites will increase explosively in the place of human daily life. The bodies and feces of mites are atomized into fine particles by drying and physical crushing and float in the air during human daily activities.

[0005] Efforts have been made to prevent exposure to these air pollutants due to their harmful effects on living organisms. Specifically, it is the use of air purifiers and wearing masks.

[0006] In a closed space such as indoors, the operation of an air purifier can be considered. However, even if an air purifier is operated at home, it is suggested to be insufficient because the floor will turn white with fine dust if the floor is not swept, or dust will accumulate on the top of the air purifier. In fact, to maintain the air cleanliness in a closed space, a large-scale device like a clean room is essential.

[0007] In open spaces such as outdoors, mask wearers are often seen in the streets. Although the microparticle passage blocking property of the mask material is expected to function as a filter, in reality, for the blocking performance of the mask material to be fully exerted, like in the case of medical N95 masks, the mask must fit perfectly against the face surface without any gaps. During inhalation, the inside of the mask becomes negative pressure, so if there is a gap between the mask and the face, air will enter through the gap with no resistance rather than through the mask surface with high resistance, and thus the blocking function of the mask will not work effectively. Also, breathing while wearing an N95 mask places a high load on the respiratory organs and is uncomfortable, so it is not suitable for long-term wear.

[0008] From the above, it is suggested that there are limitations in preventing exposure to the living body simply by filtering out air pollutants. Thus, as a method of protecting the living body from the harmfulness of air pollutants on the premise that exposure to air pollutants by the living body is inevitable, introducing an anti-inflammatory effect into the living body is considered an effective means.

[0009] Subjects who suffer health damage from outdoor-derived air pollutants include, of course, the people of our country who experience a deterioration in their physical condition during the dispersion of PM2.5 and the like, as well as the people of countries such as China, India, and Vietnam where environmental measures cannot keep up with the rapid progress of industrialization, and the people of neighboring countries. For people in many of the above countries, including our own, who complain of health damage, it is expected to contribute to health recovery. Subjects who suffer health damage from house dust, which is an indoor air pollutant, are the people of developed countries where highly airtight and highly insulated houses are increasing and emerging countries where urbanization is rapidly developing.

[0010] Although it is known that the juice and fruit peels of Citrus jabara have an anti-allergic rhinitis effect (see, for example, Patent Document 1), there has been no report on the suppression of particulate matter-induced inflammation. Further, in previous anti-inflammatory studies, lipopolysaccharide (LPS) has been used as an inflammation inducer (see, for example, Non-Patent Document 1). However, according to the studies by the present inventors, as will be described later, it has been suggested that LPS and particulate matter are greatly different in composition and also different in the inflammation induction mechanism. Therefore, from the conventional research results, it has not been suggested at all that naringenin has an effect of suppressing particulate matter-induced inflammation, nor is it a fact that can be recognized as obvious.

Prior Art Documents

Patent Documents

[0011]

Patent Document 1

Non-Patent Documents

[0012]

Non-Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0013] In view of the above situation, an object of the present invention is to provide an inhibitor for suppressing MCP-1 induced by particulate matter such as PM2.5 and yellow sand, MCP-1 production that suppresses production a food for suppression, MCP-1 production and an oral preparation for suppression. and, MCP-1 production

Means for Solving the Problems

[0014] ​To solve the above problems, the present invention has the following configuration. That is, the MCP-1 Production inhibitor according to the present invention contains naringenin and / or naringin as an active ingredient. The MCP-1 Production inhibiting food according to the present invention contains naringenin and / or naringin as an active ingredient. The MCP-1 Production oral inhibitor according to the present invention contains naringenin and / or naringin as an active ingredient.

Advantages of the Invention

[0015] The MCP-1 production inhibitor, MCP-1 production inhibiting food, and MCP-1 production oral inhibitor according to the present invention can inhibit MCP-1 Production of induced by air pollutants typified by PM2.5.

Brief Description of the Drawings

[0016]

Figure 1

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Modes for Carrying Out the Invention

[0017] Hereinafter, preferred embodiments of the floating particulate matter-induced inflammation inhibitor and the food for inhibiting floating particulate matter-induced inflammation according to the present invention will be described in detail. However, the scope of the present invention is not restricted by these descriptions, and other than the following examples, it can be appropriately modified and implemented without departing from the spirit of the present invention.

[0018] "Floating particulate matter" is generally defined as particulate matter with a particle size of 10 μm or less among the particulate matter floating in the atmosphere. Among these, those with a diameter of 2.5 μm or less are called "PM2.5". Floating particulate matter includes those from anthropogenic sources such as soot and dust discharged from factories, etc., black smoke (diesel exhaust particles: DPE) contained in the exhaust gas of diesel vehicles, etc., scattering of soil represented by yellow sand, and those from natural sources such as microorganisms such as mites and molds or their dead remains, etc., and there are actually various types. Also, when classified according to the difference in the generation mechanism, it is classified into primary particles directly discharged into the atmosphere as particles from the source of generation, and secondary particles in which substances discharged as gaseous substances are changed into particles by photochemical reactions, etc. in the atmosphere. Specific examples include PM2.5, yellow sand, atmospheric dust, combustion products of fossil fuels, DPE, house dust, mite corpses and decomposition products of feces, decomposition products of molds, etc. The floating particulate matter-induced inflammation inhibitor and the food for inhibiting floating particulate matter-induced inflammation according to the present invention suppress inflammation induced by the above-mentioned floating particulate matter.

[0019] Naringin is a kind of flavonoid, a flavanone glycoside in which the aglycone is naringenin and the sugar moiety is β-rutinose. According to the study of the present inventor, naringenin exhibits an inhibitory effect on floating particulate matter-induced inflammation. And since naringin is absorbed by colon cells after being deglycosylated to naringenin, it is considered to function like a prodrug with naringenin as the active substance in vivo. Therefore, the floating particulate matter-induced inflammation inhibitor and the food for inhibiting floating particulate matter-induced inflammation of the present invention contain naringenin, which is the active ingredient, and / or naringin that is converted into naringenin in vivo as the active ingredient.

[0020] In addition, naringin is contained in all citrus fruits such as yuzu, kabosu, grapefruit, and mandarin oranges, and is particularly abundant in the pericarp. Among citrus fruits, it is particularly abundant in yabara. Therefore, the floating particulate matter-induced inflammation inhibitor and the food for inhibiting floating particulate matter-induced inflammation of the present invention also contain yabara pericarp components as the active ingredient.

[0021] As the yabara pericarp, those peeled by hand from yabara fruits, those peeled by a known automatic peeling machine, and the residue remaining after squeezing juice with a juice press can be used. Considering the labor, the residue of the juice press is preferable. In the above, the yabara fruit can be used without particular limitation. Among them, since the content of naringin per unit weight is high, the harvested fruit or the immature fruit before the pericarp turns yellow is preferable. Specifically, the fruit picked 3 to 4 months after fruit set is preferable as the immature fruit. Since yabara generally sets fruit around June, the fruit harvested around September to October is preferable.

[0022] The yabara pericarp may be processed into jam, juice, etc. together with sugar in a raw state, for example. However, in order to enhance the storage stability, the dried pericarp is preferable. Examples of the dried pericarp include those produced by a general method, that is, a heat drying method in which the pericarp and fruit are dried in the shade, in the sun, or by a dryer as they are or after being finely cut, a freeze-drying method, a spray-drying method, or a combination of these methods.

[0023] In addition, it is preferably manufactured by drying by the heat drying method after drying by the freeze drying method, and more preferably by heat drying at 80°C. The dried peel of sea bream thus manufactured can evaporate the limonene contained in the dried peel while maintaining a high naringin concentration, and suppress its bitterness.

[0024] In addition, although this dried peel can be used as it is, it is more preferable to process it into a powder form by a general method, that is, by a ball mill or the like, in consideration of ease of eating.

[0025] The dried peel processed into a powder form can be used alone, but the taste can be improved by mixing it with furikake, the base of ochazuke, etc., or by mixing it with saccharides such as reduced maltose to form granules. In addition, the dried peel processed into granules may be contained in a capsule such as an enteric capsule to form a capsule agent, or may be mixed with known additives such as magnesium stearate and excipients, and formed into a tablet by known pharmaceutical techniques.

[0026] As the form of administration to the living body of the floating particulate matter-induced inflammation inhibitor and the food for inhibiting floating particulate matter-induced inflammation according to the present invention, absorption through the digestive tract by oral ingestion, or absorption through the epithelial cell layer by application or spraying is conceivable, but it is not limited to this.

[0027] Examples of the food for suppressing particulate matter-induced inflammation according to the present invention include general foods for humans, functional foods for health (foods for specified health use, foods with nutrient function claims), health foods, dietary supplements, and the like. Specific examples of foods include processed fishery products such as kamaboko, chikuwa, and hanpen; processed meat products such as sausage, ham, and wiener; processed agricultural products such as tofu, aburaage, and konnyaku; confectioneries; seasonings; spices; processed vegetables and fruits such as canned and bottled various vegetables and fruits; dairy products such as cheese, butter, and yogurt; fruit juices, vegetable juices, whey beverages, soft drinks, health teas, medicinal liquors, and other beverages; and other health-oriented food and drink products such as tablets, beverages, and granules for maintaining health for the purpose of nutritional reinforcement (dietary supplementation), etc. However, the present invention is not limited thereto.

[0028] Hereinafter, experimental data demonstrating the effects of the particulate matter-induced inflammation inhibitor and the food for suppressing particulate matter-induced inflammation according to the present invention are shown.

[0029] 〔Experiment 1〕 <Experimental method> Naringenin, the aglycone of flavanon glycoside naringin, which is a major component in quince peel, was purchased from KLT Laboratory and dissolved in dimethyl sulfoxide to prepare a sample. Mouse macrophage-like cell line RAW264 cells were obtained from the RIKEN BioResource Center and used after culturing in D-MEM (hereinafter abbreviated as culture medium) containing 10% fetal bovine serum, 100 units / ml of penicillin, and 100 μg / ml of streptomycin. Lipopolysaccharide (LPS) was purchased from Sigma-Aldrich. Urban air dust (NIES CRM No. 28) was obtained from the National Institute for Environmental Studies. Cells were seeded at 1.0×10 per well in a 96-well plate 5The cells were seeded with 100 cells, culture medium containing naringenin (final concentration: 0, 50, 100, 200μM) was added, and then culture medium containing LPS (final concentration: 1μg / ml) or urban air dust (final concentration: 0, 100, 200, 300, 400, 500μg / ml) was added 2 hours later. The cells were cultured at 37℃ under 5% carbon dioxide for 24 hours, and the culture supernatant was collected. The culture supernatant was centrifuged at 15,000 rpm for 10 minutes, and the concentrations of nitric oxide (NO) in the obtained supernatant were quantified using Promega's Griess reagent system, and the concentrations of IL-6, MCP-1, and TNF-α were quantified using BioLegend's ELISA kits.

[0030] <Differences from previous anti-inflammatory research> Previous studies have used lipopolysaccharide (LPS) as an inflammation inducer. LPS is a component of the outer membrane of the cell wall of gram-negative bacteria cells, and is a glycolipid composed of lipids and polysaccharides. Meanwhile, the air pollutant used in this experiment was urban air dust NIES CRM No. 28 obtained from the National Institute for Environmental Studies (air dust collected in the ventilation filters of a building in the center of Beijing over a 10-year period from 1996 to 2005, with 99% of particles 10 μm or less in diameter and 40% or more 2 μm or less in diameter). Its composition analysis, as shown in Anal Bioanal Chem (2008) 391: 1997-2003, is clearly different from LPS. The composition of the urban air dust used in the experiment is characterized by its high content of silicon (Si) at 14.9% and sulfur (S) at 3.91%. The presence of silicon (Si) suggests that there is a lot of dust components such as yellow sand, and the presence of sulfur (S) suggests that there is a lot of chemicals derived from fossil fuel combustion. The presence of a lot of fossil fuel combustion products suggests that there is also a lot of aromatic compounds and nitrogen oxides derived from fossil fuels. The inflammatory effects of LPS and urban air particulate matter were compared using nitric oxide (NO), IL-6, MCP-1, and TNF-α as indicators. LPS was used at a concentration of 1 μg / ml, as used in many studies, to stimulate mouse macrophage-like cell line RAW264 cells. As shown in Fig. 1, the NO concentration 24 hours after LPS stimulation was 11.6 μM, while that of urban air dust was 4.32 - 7.03 μM at an added amount of 100 - 500 μg / ml, showing a lower value than that at an LPS added amount of 1 μg / ml. On the other hand, for the concentrations of IL-6, MCP-1, and TNF-α 24 hours after stimulation, except for the IL-6 concentration at an added amount of 100 μg / ml of urban air dust, the urban air dust added system far exceeded the value at an LPS added amount of 1 μg / ml and showed a value more than five times that at the time of LPS addition. Since urban air dust is a mixture of various components, it is expected that there are also various inflammatory inducers, and this result is considered reasonable. From the above, it was suggested that both LPS and urban air dust showed an inflammatory induction effect, but not only were their compositions greatly different, but their inflammatory induction mechanisms were also different. Since the inflammatory induction mechanisms of action are different, there is no guarantee that the anti-inflammatory effect observed in the LPS added system will be similarly exerted in the urban air dust added system, and it is considered necessary to measure and evaluate it actually.

[0031] <Suppressive Effect of Naringenin on Urban Air Dust-Induced Inflammatory Response> As shown in Fig. 1, naringenin dose-dependently suppressed the induction of the production of NO, IL-6, and MCP-1 by the addition of urban air dust (added amount 100 - 500 μg / ml). From this, it was suggested that naringenin has an effect of suppressing the inflammatory induction by urban air dust. However, with regard to TNF-α, almost no inhibitory effect was observed, which was different from the result at the time of LPS stimulation (see Non-Patent Document 1). This may also mean that it is due to the difference in the inflammatory induction mechanisms of LPS and urban air dust.

[0032] [Experiment 2] Gobi dust NIES CRM No.30 obtained from the National Institute for Environmental Studies (a dust sample containing sand and dust separated and collected by aerodynamic methods based on surface soil containing yellow sand collected from the yellow sand source area in the southwestern Gobi Desert. In May 2011, during the yellow sand generation season, surface soil containing 1200 kg of yellow sand was collected at four locations near 44°N and 109°E. Using a dust generation chamber installed in the management facility within the Sainshand Meteorological Observation Center, dust was generated from the surface soil. Only fine dust was separated and collected using a cyclone type classifier (separation limit diameter 10 μm), and approximately 2 kg of secondary raw material was obtained. Subsequently, at the National Institute for Environmental Studies, purification separation and homogenization by a rotary blender were performed to obtain 1.2 kg of the final raw material, 60 which was sterilized by Co irradiation (25 kGy) and used. As described in Analytical Methods, 2013, 5, 4088 - 4095, its composition shows that silicon (Si) accounts for 24.1%, while sulfur (S) is estimated to be below the detection limit. Under the condition of a yellow sand concentration of 100 μg / ml, similar to the experiment with LPS and urban airborne dust (Experiment 1), it was added to the mouse macrophage cell line RAW264 cells to evaluate the effect of naringenin. The results are shown in Figure 2. Naringenin showed an effective inhibitory effect on NO production and IL - 6 production in a concentration - dependent manner, similar to when urban airborne dust was added. However, the inhibitory effect on MCP - 1 and TNF - α was slow. Since the inflammation - inducing effect of yellow sand and the inhibitory effect of naringenin are different from those of LPS, there is no guarantee that the anti - inflammatory effect of naringenin in conventional LPS stimulation will be similarly exerted in yellow sand stimulation, and it is considered necessary to evaluate by actual measurement.

[0033] [Experiment 3] Using human airway epithelial cell line BEAS - 2B cells (purchased from DS Pharma Biomedical Co., Ltd.) and BEGM medium (purchased from LONZA) as the culture medium, similar to the experiment with RAW264 cells, LPS, urban airborne dust, and yellow sand were used as inflammation - inducing stimulants, and the anti - inflammatory effect of naringenin (concentrations: 0, 100, 200 μM) was evaluated by its inhibitory effect on IL - 6 production. The results are shown in Figure 3. Urban airborne dust did not show an effect of inducing IL-6 production in BEAS-2B cells. LPS and yellow sand induced IL-6 production, and an inhibitory effect was observed in a concentration-dependent manner with the addition of naringenin.

[0034] [Experiment 4] Extracts of mite bodies obtained from Biosuta Co., Ltd. were added to mouse macrophage cell line RAW264 cells under the conditions of final concentrations of 100, 50, 10, and 5 μg / ml, in the same manner as the experiments with LPS and urban airborne dust (Experiment 1), to evaluate the effect of naringenin. [Suppressive effect of naringenin on mite body extract-induced inflammatory response] As shown in Fig. 4, naringenin dose-dependently suppressed the induction of production of NO, IL-6, and MCP-1 by the addition of mite body extracts (added amount 5 - 100 μg / ml). From this, it was suggested that naringenin has an effect of suppressing inflammation induction by mite body extracts. However, with regard to TNF-α, almost no inhibitory effect was observed, which was different from the results during LPS stimulation (see Non-Patent Document 1). This may mean that it is due to the difference in the inflammation induction mechanisms of LPS and mite body extracts.

[0035] [Experiment 5] Extracts of mite feces obtained from Biosuta Co., Ltd. were added to mouse macrophage cell line RAW264 cells under the conditions of final concentrations of 100, 50, 10, and 5 μg / ml, in the same manner as the experiments with LPS and urban airborne dust (Experiment 1), to evaluate the effect of naringenin. [Suppressive effect of naringenin on mite feces extract-induced inflammatory response] As shown in Fig. 5, naringenin dose-dependently suppressed the induction of production of NO, IL-6, and MCP-1 by the addition of mite body extracts (added amount 5 - 100 μg / ml). From this, it was suggested that naringenin has an effect of suppressing inflammation induction by mite body extracts. However, with regard to TNF-α, almost no inhibitory effect was observed, which was different from the results during LPS stimulation (see Non-Patent Document 1). This may mean that it is due to the difference in the inflammation induction mechanisms of LPS and mite body extracts. In addition, since the degree of the inflammation-inducing effect and the inhibitory effect by naringenin differ between the mite extract and the worm body extract and the fecal extract, there is no guarantee that the anti-inflammatory effect of naringenin in conventional LPS stimulation will be similarly exerted in mite stimulation, and it is considered necessary to evaluate by actual measurement.

Claims

1. An MCP-1 production inhibitor containing naringenin and / or naringin as an active ingredient.

2. A food for inhibiting MCP-1 production containing naringenin and / or naringin as an active ingredient.

3. An oral preparation for inhibiting MCP-1 production containing naringenin and / or naringin as an active ingredient.

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