KR102703034B1 - Composition for preventing, improving or treating pulmonary fibrosis comprising complex of anthocyanin and anionic polysaccharide - Google Patents

Abstract

The present invention relates to a composition for preventing, improving or treating pulmonary fibrosis, comprising an anthocyanin-negatively charged polysaccharide complex as an effective ingredient, and more specifically, to a pharmaceutical composition for preventing, improving or treating pulmonary fibrosis, comprising an anthocyanin-negatively charged polysaccharide complex as an effective ingredient, which exhibits a synergistic effect compared to each of anthocyanin and negatively charged polysaccharide substances.
The composition according to the present invention can exhibit excellent effects in preventing or treating pulmonary fibrosis by regulating the cell transmission signal system of TGF-β, which directly affects the development of pulmonary fibrosis.

Description

{Composition for preventing, improving or treating pulmonary fibrosis comprising complex of anthocyanin and anionic polysaccharide as active ingredient}

The present invention relates to a composition for preventing, improving or treating pulmonary fibrosis, comprising an anthocyanin-negatively charged polysaccharide complex as an effective ingredient, and more specifically, to a pharmaceutical composition for preventing, improving or treating pulmonary fibrosis, comprising an anthocyanin-negatively charged polysaccharide complex as an effective ingredient, which exhibits a synergistic effect compared to each of anthocyanin and negatively charged polysaccharide substances.

The lungs are essential organs responsible for human breathing, obtaining oxygen and expelling carbon dioxide through inhalation and exhalation of air. Pulmonary fibrosis is a disease that worsens this important lung function and can even lead to death. It is a disease in which the parenchymal cells of the lungs become fibrosed and gradually contract and harden due to frequent recurrence of inflammation in the lungs. In other words, acute inflammation caused by bacterial or viral infection (pulmonary tuberculosis, cold, influenza, new influenza, etc.) becomes chronic and the inflammation remains, or the sympathetic nerves that affect immune activity continue to be hyperactive, causing an imbalance in the autonomic nervous system and excessive immune activity, and the tissues where this inflammation occurs repeatedly gradually harden and progress to pulmonary fibrosis.

Currently, steroids that suppress inflammation are used as a treatment for pulmonary fibrosis, but these only temporarily suppress inflammation and do not have a therapeutic effect on inflammation that has already occurred. In addition, long-term use causes various side effects, such as weakening of the skin and bones and deformation of blood vessels. Therefore, the development of a treatment that can effectively treat pulmonary fibrosis is urgently needed.

Accordingly, the inventor of the present invention conducted extensive research to develop a natural product-based pulmonary fibrosis treatment agent, and as a result, discovered that a complex of anthocyanin and negatively charged polysaccharide exhibited a significantly improved pulmonary fibrosis prevention or treatment effect compared to treatment with each substance alone, thereby completing the present invention.

Accordingly, the purpose of the present invention is to provide a pharmaceutical composition for preventing or treating pulmonary fibrosis, which comprises a complex of anthocyanin and negatively charged polysaccharide as an effective ingredient.

Another object of the present invention is to provide a food composition for preventing or improving pulmonary fibrosis, which comprises a complex of anthocyanin and negatively charged polysaccharide as an effective ingredient.

Another object of the present invention is to provide a health functional food or health supplement containing the food composition.

In order to achieve the above-mentioned object of the present invention, the present invention provides a pharmaceutical composition for preventing or treating pulmonary fibrosis, which comprises a complex of anthocyanin and negatively charged polysaccharide as an effective ingredient.

In order to achieve another object of the present invention, the present invention provides a food composition for preventing or improving pulmonary fibrosis, which comprises a complex of anthocyanin and negatively charged polysaccharide as an effective ingredient.

In order to achieve another object of the present invention, the present invention provides a health functional food or health supplement containing the food composition.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for preventing or treating pulmonary fibrosis, comprising a complex of anthocyanin and negatively charged polysaccharide as an effective ingredient.

Aronia (Aronia melanocarpa) is a berry belonging to the Rosaceae family, native to Northeastern America and Eastern Europe. Aronia, which is dark red in color, has recently been widely cultivated around the world, and is cultivated by crossbreeding two varieties in horticulture. In particular, Aronia is known to belong to the group with the highest anthocyanin among natural plants. Aronia contains many types of anthocyanins such as Cyanidin-3-galactoside, Cyanidin-3-glucoside, Cyanidin-3-arabinoside, and Delphinidin-3-glucoside, and is also known to contain a large amount of phenol. Such Aronia has been shown to have excellent antioxidant effects, anti-inflammation, and immunity enhancement effects through research, and many studies have been conducted on its effects on diabetes, various cardiovascular diseases, and cosmetics.

In the present invention, the 'anthocyanin' refers to a plant-based natural pigment that exhibits colors such as purple, red, and blue depending on the acid concentration of the cell fluid, the chemical structure of the pigment compound, and the state of bonding with various metal ions, which is a water-soluble pigment glycoside present in the plant. Recently, it has been known that anthocyanin has various physiological activities, and for example, its anti-aging effect, anti-bacterial effect, anti-mutagenic effect, cholesterol-lowering effect, vision-improving effect, vascular protection function, and anti-ulcer function have been elucidated.

In the present invention, the anthocyanin may be isolated from a plant. The plant includes any plant that produces anthocyanin, and may be, for example, black rice, black bean, black currant, chokeberry, black chokeberry, cranberry, aronia, mulberry, cherry, raspberry, blueberry, blackberry, eggplant, acai, cornelian cherries, or grapes, and preferably, aronia.

In the present invention, the anthocyanin may be obtained by crushing black rice, black beans, black currant, chokeberry, black chokeberry, cranberry, aronia, mulberry, cherry, raspberry, blueberry, blackberry, eggplant, acai, cornelian cherry or grape, adsorbing the juice onto a polyphenol adsorption resin, and eluting the same with an alcohol having 1 to 5 carbon atoms.

In the present invention, the anthocyanin is peonidin, cyanidin 3-arabinoside, cyanidin-3-(xylosylglucose)-5-galactose, cyanidin 3-glucoside, cyaniding-3-xyloside, cyanidin 3-galactoside, cyanidin-3-(coumaroyl-xylosylglucose)-5-galactose, delphinidin 3-glucoside, delphinidin 3-rutinoside. 3-rutinoside), peonidin 3-arabinoside, peonidin 3-galactoside, petunidin 3-glucoside, cyanidin, delphinidin, malvidin, pelargonidin, peonidin, cyanidin 3,5-diglucoside, cyanidin 3-rutinoside, pelargonidin 3-glucoside, peonidin 3-glucoside, malvidin 3-glucoside and malvidin It may include at least one anthocyanin selected from the group consisting of malvidin 3,5-diglucoside, preferably cyanidin-3-galactoside, cyanidin-3-glucoside, cyaniding-3-arabinoside, and cyaniding-3-xyloside.

In the present invention, the negatively charged polysaccharide is not particularly limited as long as it is biocompatible, and may include one or more types of negatively charged functional groups. The term 'polysaccharide' refers to a polymer of two or more monosaccharide molecules, wherein the monosaccharides may be the same or different. The polysaccharide of the present invention may or may not be cross-linked.

The polysaccharide used to form the complex of the present invention can be formed by using the negatively charged polysaccharide itself, or by introducing a chemical formula to a general polysaccharide containing no or a small amount of negatively charged functional groups to impart negatively charged functional groups. The negatively charged functional groups can be, for example, one or two or more carboxyl groups or sulfate groups.

In a specific example, the negatively charged polysaccharide can be selected from the group consisting of hyaluronic acid, o-sulfated hyaluronic acid (o-sulfated HA), dextran sulfate, chondroitin sulfate, dermatan sulfate, keratin sulfate, heparin, heparin sulfate, alginic acid, fucoidan, carrageenan, mixtures thereof and complexes thereof, preferably fucoidan or alginic acid, and most preferably fucoidan, but is not limited thereto.

The above negatively charged polysaccharide is a substance frequently used in foods and beverages, and is not only harmless to the human body, but also has a strong negative charge in a neutral solution, so it can effectively form an ion bond with positively charged anthocyanins to form a complex.

The polysaccharide described above has no particular limitation, but may have a weight average molecular weight in the range of about 1,000 to 1,000,000, preferably in the range of about 10,000 to 300,000, and more preferably in the range of 20,000 to 50,000.

Since the negatively charged polysaccharide of the present invention can control the loading rate of anthocyanin by controlling the ionic bonding ratio with anthocyanin using the type and number of existing negative charges of the polysaccharide, more than one type and polysaccharide of various molecular weights can be used depending on the purpose as long as it is a negatively charged polysaccharide.

In the above complex, the weight ratio of anthocyanin and negatively charged polysaccharide may be 1:1 to 100, preferably 1:1 to 50, more preferably 1:1 to 30, even more preferably 1:1 to 20, and most preferably 1:1 to 10. Within the above range, the complex formed from the polysaccharide of the present invention may exhibit advantageous effects in terms of enhancing the stability and physiological activity of anthocyanin.

More specifically, the weight ratio of anthocyanin to negatively charged polysaccharide in the complex can be 1:1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20.

Anthocyanins are known to have high stability at low pH, and in particular, have the highest stability and the highest antioxidant activity at pH 3 or lower. In addition, since they have a positive charge at pH 3 or lower, they can form a complex through ionic bonding with biocompatible negatively charged polysaccharides, thereby becoming structurally stable. Therefore, anthocyanin-negatively charged polysaccharide nanocomplexes have excellent oxidation stability and storage stability, and thus can improve processing conditions, distribution, and storage properties when applied to drugs or foods. In addition, since enhanced stability can enhance absorption in the body, they can be very useful for pharmaceutical development.

In addition, in the present invention, the complex may have a size ranging from 10 nm to 1000 μm after manufacturing by forming a complex of anthocyanin and negatively charged polysaccharide, and may be in a form in which anthocyanin is encapsulated by the polysaccharide, but is not limited thereto. The anthocyanin-negatively charged polysaccharide complex according to the present invention may be in a suspension or powder state depending on the purpose of use or the field of application.

In some cases, the anthocyanin-negatively charged polysaccharide complex may further comprise a biocompatible or biodegradable carrier. In this case, the anthocyanin-negatively charged polysaccharide complex is within or is a part of the carrier, and the carrier may be a liposome, a micelle, or a polymerized vesicle.

In the present invention, the most preferable form of the complex is that the negatively charged polysaccharide is alginic acid or fucoidan having a weight average molecular weight of 20,000 to 50,000, and the anthocyanin is a cyanidin glycoside, preferably cyanidin-3-galactoside, cyanidin-3-glucoside, cyanidin-3-arabinoside and/or cyanidin-3-xyloside.

The above cyanidin glycosides can form a complex by ionic bonding with negatively charged polysaccharides.

In the present invention, the anthocyanin-negatively charged polysaccharide complex may be manufactured by a method including the following steps, although the manufacturing method thereof is not particularly limited:

(i) a step of forming a complex of anthocyanin and negatively charged polysaccharide in a neutral or acidic environment; and/or

(ii) A step of recovering the complex formed above.

In the above, when forming a complex of anthocyanin and negatively charged polysaccharide, there is no particular limitation on the solvent, but the more edible it is, the better. After preparing the negatively charged polysaccharide and anthocyanin in an appropriate ratio, the two solutions can be stirred evenly and waited for the complex to be completely formed at an appropriate temperature.

The complex formed by the method (i) above may be maintained in an acidic environment as is if it is an oral formulation, depending on the formulation, but if it is used as an injection, it may be desirable to neutralize it to a neutral pH by the method (ii).

In the present invention, the "pulmonary fibrosis" refers to a type of chronic interstitial lung disease, also known as idiopathic pulmonary fibrosis, in which lung tissue cells change into fibrocytes, causing dyspnea, cough, cyanosis, clubbing, etc. When examining a tissue, honeycomb-shaped or atypical fibrocyte clusters are observed. Up to now, steroid-based therapeutic agents, interferon gamma, acetylcysteine, pirfenidone, bocetan, etc. have been used as therapeutic agents, but no agent showing a specific therapeutic effect has been reported.

In the present invention, the cause of the pulmonary fibrosis is not limited, and non-limiting examples thereof may include idiopathic pulmonary fibrosis, pulmonary fibrosis due to anticancer treatment, pulmonary fibrosis due to MERS or pneumonia, etc.

Meanwhile, the term 'comprising as an active ingredient' in the present specification means comprising a sufficient amount to achieve the efficacy or activity of the anthocyanin-negatively charged polysaccharide complex. In one specific embodiment of the present invention, the anthocyanin-negatively charged polysaccharide complex in the composition of the present invention is comprised, for example, at least 0.001 mg/kg, preferably at least 0.1 mg/kg, more preferably at least 10 mg/kg, still more preferably at least 100 mg/kg, still more preferably at least 250 mg/kg, and most preferably at least 0.1 g/kg. Since the anthocyanin-negatively charged polysaccharide complex is a natural product and does not have side effects on the human body even when administered in excessive amounts, a person skilled in the art can select and implement the quantitative upper limit of the anthocyanin-negatively charged polysaccharide complex included in the composition of the present invention within an appropriate range.

The pharmaceutical composition of the present invention can be prepared using pharmaceutically suitable and physiologically acceptable excipients in addition to the effective ingredient, and the excipients can include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, glidants, or flavoring agents.

The above pharmaceutical composition may be preferably formulated as a pharmaceutical composition by additionally including one or more pharmaceutically acceptable carriers in addition to the effective ingredients described above for administration.

The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an orally, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. In addition, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture, if desired or necessary. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweetener, acacia, tracheacanth or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, etc.

In the composition formulated as a liquid solution, acceptable pharmaceutical carriers are sterile and biocompatible, and include saline solution, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and one or more of these components can be mixed and used. As necessary, other conventional additives such as antioxidants, buffers, and bacteriostatic agents can be added. In addition, diluents, dispersants, surfactants, binders, and lubricants can be additionally added to formulate the composition as an injectable formulation such as an aqueous solution, suspension, or emulsion, or as a pill, capsule, granule, or tablet.

Furthermore, it can be preferably formulated according to the ingredient using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA, as an appropriate method in the field.

The pharmaceutical composition of the present invention can be administered orally or parenterally, and in the case of parenteral administration, can be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc., and oral administration is preferred.

The suitable dosage of the pharmaceutical composition of the present invention varies depending on factors such as the formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity, and a skilled physician can easily determine and prescribe a dosage effective for the desired treatment or prevention. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-10 g/kg.

The pharmaceutical composition of the present invention can be manufactured in a unit dose form or can be manufactured by placing it in a multi-dose container by formulating it using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily performed by a person having ordinary skill in the art to which the present invention pertains, and thereby. At this time, the formulation may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granules, tablets or capsules, and may additionally contain a dispersant or stabilizer.

The present invention also provides a food composition for preventing or improving pulmonary fibrosis, comprising a complex of anthocyanin and negatively charged polysaccharide as an effective ingredient.

The food composition according to the present invention can be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gum, ice cream, health supplements (beverages, low viscosity gel types, pills, tablets, capsules, etc.), vitamin complexes, etc.

The food composition of the present invention may contain not only the anthocyanin-negatively charged polysaccharide complex as an effective ingredient, but also components commonly added during food manufacturing, such as proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents. Examples of the above-mentioned carbohydrates include monosaccharides such as glucose, fructose, etc.; disaccharides such as maltose, sucrose, oligosaccharides, etc.; and polysaccharides such as dextrin, cyclodextrin, etc., and conventional sugars and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [thaumatin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.]) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is manufactured into a drink or beverage, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts may be additionally included in addition to the anthocyanin-negatively charged polysaccharide complex of the present invention.

The present invention relates to a health functional food comprising the food composition comprising the anthocyanin-negatively charged polysaccharide complex as an effective ingredient. Or, a health supplement is provided. A health functional food or health supplement refers to a food manufactured by adding an anthocyanin-negatively charged polysaccharide complex to food materials such as beverages, teas, spices, gums, and confectionery, or by manufacturing it in the form of encapsulation, powder, suspension, etc., and means that when consumed, it has a specific health effect, but unlike general drugs, it has the advantage of not having side effects that may occur when taking drugs for a long period of time because it uses food as a raw material. The health functional food or health supplement of the present invention obtained in this way is very useful because it can be consumed on a daily basis. The amount of the anthocyanin-negatively charged polysaccharide complex added to such health functional foods or health supplements varies depending on the type of the target health functional food and cannot be uniformly specified, but it may be added within a range that does not damage the original taste of the food, and is usually in the range of 0.01 to 50 wt%, preferably 0.1 to 20 wt%, with respect to the target food. In addition, in the case of health functional foods or health supplements in the form of pills, granules, tablets or capsules, it is usually added in the range of 0.1 to 100 wt%, preferably 0.5 to 80 wt%. In one specific example, the health functional foods or health supplements of the present invention may be in the form of pills, tablets, capsules or beverages.

The composition according to the present invention can exhibit excellent effects in preventing or treating pulmonary fibrosis by regulating the cell transmission signal system of TGF-β, which directly affects the development of pulmonary fibrosis.

Figure 1 shows the results of Western blot analysis of the phosphorylation levels of Smad2 and Smad3, which are downstream signaling steps of TGF-β, after treating A549 cells with TGF-β and each test substance (ABF, CFNC4, CFNC8) (1A) and the results of quantifying the results (1B).
Figure 2 shows the results of evaluating the degree of cell migration after creating an environment similar to the early stage of pulmonary fibrosis by treating A549 cells with TGF-β and then treating each test substance (ABF, CFNC4, CFNC8) for 24 or 48 hours.
Figure 3 is a graph showing the quantification of the degree of cell movement according to Figure 2.

Hereinafter, the present invention will be described in detail by the following examples. However, the following examples are only intended to illustrate the present invention, and the present invention is not limited thereto.

Example 1: Preparation of anthocyanin-negatively charged polysaccharide complex material

Anthocyanin was obtained by crushing the aronia berries, adsorbing the juice to a polyphenol adsorption resin, eluting it with an aqueous ethyl alcohol solution, and powdering it (hereinafter referred to as 'ABF'). The anthocyanin powder thus prepared was dissolved in 5 ml of phosphate buffer pH 3 at 40°C, and fucoidan, a negatively charged polysaccharide, was dissolved in 10 ml of deionized water (DI water) at room temperature. The anthocyanin solution was mixed with the fucoidan solution such that the weight ratio of fucoidan:anthocyanin contained in the solution was 1 to 6:1 (CFNC4) or 6 to 20:1 (CFNC8), and then stirred at room temperature for 48 hours and freeze-dried to prepare an anthocyanin-negatively charged polysaccharide complex (CFNC4 and CFNC8, respectively).

The above anthocyanin-negatively charged polysaccharide complex was prepared as a solution at a concentration of 10 mg/ml using cell culture medium (Ham's F-12K; 10% FBS, 1% Antibiotics) prior to use in the test, and then diluted using the same culture medium to the concentration required for each test and used.

Example 2: Analysis of TGF-β signaling pathway proteins

In the early stage of pulmonary fibrosis, an epithelial to mesenchymal transition (EMT) process occurs, in which epithelial cells transform into mesenchymal cells, and EMT occurs by the induction of TGF-β. Therefore, the prevention of pulmonary fibrosis can be predicted through the presence or absence of phosphorylation of Smad2 and Smad3, which are downstream signaling steps of TGF-β. In this study, the amount of phosphorylation of Smad2 and Smad3 induced by TGF-β was measured by performing a western blot assay. The phosphorylated form (activated form) of Smad2/3 is expressed as p-Smad2 and p-Smad3 below. The formation of p-Smad2 and p-Smad3 indicates the possibility of pulmonary fibrosis development.

The experimental method is as follows:

1) 24 hours before treatment with test substances (ABF, CFNC4, CFNC8), subculture A459 cells using Ham's F-12K (10% FBS, 1% Antibiotics) medium.

2) Treat with 0.1 mg/ml of the test substance and 46 hours later, treat with 5 ng/ml of TGF-β for 2 hours.

3) 48 hours after treatment with the test substance, cells are collected, lysed, and quantified using the Bradford assay.

4) Analyze the same amount of protein using a western blot assay.

The results are shown in Figure 1.

Referring to Figure 1, the results of this test confirmed that when the substances CFNC4 and CFNC8 were treated, the phosphorylation formation of Smad2 and Smad3 was reduced (Figure 1A), and this result was verified through statistical processing through repeated experiments (Figure 1B). Statistical analysis was performed using the graph pad Prism5 program, and one-way ANOVA and post-hoc analysis were performed with Bonferroni's test.

Example 3: Cell migration assay (migration assay, wound healing assay)

The potential of the test substances (ABF, CFNC4, CFNC8) to prevent and treat pulmonary fibrosis was confirmed through migration assays that examine cell mobility and cell-to-cell binding.

The experimental method is as follows:

1) 24 hours before treatment with test substances (ABF, CFNC4, CFNC8), subculture A459 cells using Ham's F-12K (10% FBS, 1% Antibiotics) medium.

2) After 24 hours, draw lines on the cells at equal intervals using the tip.

3) After co-treatment with 0.1 mg/ml of test substance and 5 ng/ml of TGF-β, cell mobility was observed at 24 and 48 hours.

The experimental results are shown in Figures 2 and 3.

As shown in Figures 2 and 3, when the test substances CFNC4 and CFNC8 were treated for 24 hours, mobility was inhibited compared to the group treated with only TGF-β. Statistical analysis was performed using one-way ANOVA and post-hoc Bonferroni's test.

The composition according to the present invention can exhibit excellent effects in preventing or treating pulmonary fibrosis by regulating the cell transmission signal system of TGF-β, which directly affects the development of pulmonary fibrosis, and thus has high potential for industrial use.

Claims (9)

A pharmaceutical composition for preventing or treating pulmonary fibrosis, comprising a complex of anthocyanins and fucoidan derived from Aronia as an effective ingredient,
A pharmaceutical composition for preventing or treating pulmonary fibrosis, characterized in that the weight ratio of aronia-derived anthocyanin and fucoidan forming a complex is 1:1 to 20.
delete delete A pharmaceutical composition in claim 1, wherein the anthocyanin comprises at least one anthocyanin selected from the group consisting of cyanidin 3-arabinoside, cyanidin-3-xyloside, cyanidin 3-glucoside, and cyanidin 3-galactoside.
In claim 1, a pharmaceutical composition characterized in that the complex of anthocyanin and fucoidan derived from aronia is prepared by a method comprising the following steps:
(a) a step of forming a complex of anthocyanin and fucoidan derived from aronia under neutral or acidic conditions; and
(b) A step of recovering the formed complex.
delete A pharmaceutical composition according to claim 1, characterized in that the composition inhibits pulmonary fibrosis by inhibiting phosphorylation of smad2 and smad3.
A health functional food composition for preventing or improving pulmonary fibrosis, comprising a complex of aronia-derived anthocyanins and fucoidan as effective ingredients,
A health functional food composition for preventing or improving pulmonary fibrosis, characterized in that the weight ratio of aronia-derived anthocyanin and fucoidan forming a complex is 1:1 to 20.

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