Composition for preventing, treating or ameliorating inflammatory diseases

Abstract

The pharmaceutical composition for preventing or treating inflammatory diseases according to the present invention contains marigold and basil extracts as active ingredients. In addition, the functional food composition for preventing or improving inflammatory diseases of the present invention contains marigold and basil extracts as active ingredients.

Description

Technical Field

[0001] The present invention relates to a composition for preventing, treating or improving inflammatory diseases. More specifically, the present invention relates to a composition using plant materials, which can effectively prevent, treat or improve inflammatory diseases by regulating the expression of inflammatory disease-related factors. Background Art

[0002] The inflammatory response is a type of biological response to random stimuli such as tissue damage or microbial infection and is essential for normal body maintenance. However, when such an inflammatory response occurs excessively, the tissues or organs of the body in the inflamed area cannot maintain their original functions, thereby causing pain and fever. Diseases caused by excessive inflammatory responses are called inflammatory diseases. Although a large number of studies have been conducted on anti-inflammatory agents for curing these inflammatory diseases, there is still a need for safer and more effective anti-inflammatory agents.

[0003] A representative inflammatory disease is arthritis. Arthritis is a disease in which damage or inflammation occurs in joints at the bone intersections due to various reasons. Many people suffer from arthritis, which may have an adverse impact on social activities, especially by hindering the mobility of patients. In particular, degenerative arthritis in arthritis is a progressive and irreversible degenerative disease that reduces the quality of life and is the most common degenerative disease affecting 8 / 10 of the elderly. In particular, degenerative arthritis is the most representative degenerative disease, which causes pain, impaired activities of daily living and movement disorders, thereby reducing the quality of life and consuming astronomical social treatment costs, etc. However, there is currently no therapeutic agent for it.

[0004] Degenerative arthritis is a disease that causes degeneration of articular cartilage tissue and changes in the subchondral bone structure. Its pathological causes have not been clearly identified, but the main causes include cartilage wear due to repeated use during the aging process, genetics, damage to cartilage tissue caused by impact, pressure caused by overweight, and weakening of the muscles around the knee. In particular, congenital factors (such as aging and genetics) and mechanical factors (such as obesity, injury and muscle weakening) activate the biochemical pathways leading to degenerative arthritis.

[0005] The present inventors have discovered effective materials for the above-mentioned inflammatory diseases from plant materials to develop pharmaceutical compositions and functional food compositions that can effectively prevent, treat or improve arthritis, especially degenerative arthritis, and have confirmed that the combination of marigold and basil is very effective for these diseases. Therefore, the present invention has been completed. Summary of the Invention

[0006] Technical problem

[0007] The present invention relates to providing a pharmaceutical composition and a functional food composition which can effectively prevent, treat or improve inflammatory diseases such as arthritis, especially degenerative arthritis, by using plant materials.

[0008] Technical solution

[0009] One aspect of the present invention provides a pharmaceutical composition for preventing or treating inflammatory diseases, the pharmaceutical composition comprising marigold and basil extracts as active ingredients.

[0010] Another aspect of the present invention provides a functional food composition for preventing or improving inflammatory diseases, the functional food composition comprising marigold and basil extracts as active ingredients.

[0011] Beneficial effect

[0012] The composition of the present invention uses plant materials and can effectively prevent, treat or improve inflammatory diseases such as arthritis, especially degenerative arthritis, based on the inhibitory effect on the increased expression of related factors (such as MMP3, MMP13, IL-6 and COX-2) caused by inflammatory responses. In particular, the composition of the present invention can exhibit a synergistic effect from the combination of two types of plant materials. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 Shows the expression levels of MMP3, MMP13, IL-6 and COX-2 obtained by qRT-PCR when chondrocytes isolated from mouse knee joints were treated with IL-1β (1 ng / mL) to induce degenerative arthritis and then treated with a mixture of marigold extract and basil extract at a weight ratio of 1:1 (marigold + basil), marigold extract alone (marigold), and basil extract alone (basil). *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, n.s. = not significant.

[0014] Figure 2 Shows the expression levels of MMP3, MMP13, IL-6 and COX-2 obtained by qRT-PCR when chondrocytes isolated from mouse knee joints were treated with IL-1β (1 ng / mL) to induce degenerative arthritis and then treated with a mixture of marigold extract and basil extract at a weight ratio of 4:1, 2:1, 1:0.1 or 1:2. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, n.s. = not significant.

[0015] Figure 3Shows the results of observing the therapeutic effects by the concentration-dependent administration of the experimental material in mice with osteoarthritis induced by destabilization of the medial meniscus (DMM). Figure 3 Shows photographs of the joints, cartilage, and synovium stained with Safranin O after the experimental material was administered at different concentrations to each of the mice in the normal group (Sham) and the mice with DMM-induced osteoarthritis for 10 weeks and the mice were dissected. [PBS, negative control administered phosphate-buffered saline (PBS); WGA-M001, group administered a complex of Tagetes erecta extract and Ocimum basilicum extract; Tagetes erecta, group administered only Tagetes erecta extract; Ocimum basilicum, group administered only Ocimum basilicum extract; Boswellia, positive control administered Boswellia extract (currently used as a functional raw material for joint health); and Perna canaliculus, positive control administered Perna canaliculus extract (currently used as a functional ingredient for joint health)].

[0016] Figure 4 Is a graph showing the grades of osteoarthritis according to the concentration-dependent administration of the experimental material in mice with DMM-induced osteoarthritis evaluated by the OARSI system. [Sham, normal group mice; DMM, mice with DMM-induced osteoarthritis; PBS, negative control administered PBS; WGA-M001, group administered a complex of Tagetes erecta extract and Ocimum basilicum extract; Tagetes erecta, group administered only Tagetes erecta extract; Ocimum basilicum, group administered only Ocimum basilicum extract; Boswellia, positive control administered Boswellia extract; Perna canaliculus, positive control administered Perna canaliculus extract]. Values are expressed as mean ± SD, and were evaluated using the Kruskal-Wallis test and the Mann-Whitney U test. Significant differences from the negative control (PBS) are indicated as *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, and n.s = not significant.

[0017] Figure 5It is a graph showing the subchondral bone plate (SBP) thickness administered in a concentration-dependent manner of experimental materials in mice with DMM-induced osteoarthritis. [Sham, normal group mice; DMM, mice with DMM-induced osteoarthritis; PBS, negative control administered PBS; WGA-M001, group administered a complex of marigold extract and basil extract; Marigold, group administered only marigold extract; Basil, group administered only basil extract; Boswellia, positive control administered Boswellia extract; Green-lipped mussel, positive control administered green-lipped mussel extract]. Values are expressed as mean ± SD and were evaluated using the Kruskal-Wallis test and the Mann-Whitney U test. Significant differences from the negative control (PBS) are indicated as *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, and n.s = not significant.

[0018] Figure 6 It is a graph evaluating the maturity (degree of formation) of osteophytes administered in a concentration-dependent manner of experimental materials in mice with DMM-induced osteoarthritis. [Sham, normal group mice; DMM, mice with DMM-induced osteoarthritis; PBS, negative control administered PBS; WGA-M001, group administered a complex of marigold extract and basil extract; Marigold, group administered only marigold extract; Basil, group administered only basil extract; Boswellia, positive control administered Boswellia extract; Green-lipped mussel, positive control administered green-lipped mussel extract]. Values are expressed as mean ± SD and were evaluated using the Kruskal-Wallis test and the Mann-Whitney U test. Significant differences from the negative control (PBS) are indicated as *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, and n.s = not significant. Detailed Description

[0019] The composition of the present invention is characterized by containing marigold and basil extracts as active ingredients.

[0020] Marigold is an annual plant belonging to the genus Tagetes of the family Asteraceae and is mainly used as tea or as a flavoring or coloring agent for food. In the present invention, for example, one marigold species selected from Tagetes erecta, T. patula, T. minuta, and T. lucida can be used, preferably Tagetes erecta. Additionally, in the present invention, the above-ground part of the marigold can be used as the extraction raw material.

[0021] Basil is an annual plant belonging to the Lamiaceae family and is mainly used in Italian and French cuisine. In the present invention, for example, a basil species selected from sweet basil (Ocimum basilicum), holy basil (O. tenuiflorum), American basil (O. americanum), Amazon basil (Ocimum campechianum), and African basil (O. gratissimum) is used, preferably sweet basil. Additionally, in the present invention, the above-ground part of the basil can be used as an extraction raw material.

[0022] The term "extract" as used herein may refer to an extraction product, including a liquid component obtained by immersing an extraction raw material in a solvent and extracting for a certain period of time at room temperature or under heating, or a solid content obtained by removing the solvent from the liquid component. In addition to the obtained product, the extract can also be comprehensively interpreted to include its dilutions and concentrated solutions, as well as materials obtained by purifying the above product.

[0023] The extract of the present invention can be prepared using marigold and basil as extraction raw materials according to extraction methods commonly used in the art. For example, extraction can be carried out using a polar solvent or a non-polar solvent. The polar solvent can be selected from the group consisting of C1 to C4 lower alcohols and water, and the non-polar solvent can be selected from the group consisting of chloroform, petroleum ether, diethyl ether, hexane, toluene, ethyl acetate, and dichloromethane. For the effects to be achieved in the present invention, the extract of the present invention is preferably obtained using water, ethanol, or a mixed solution thereof as a solvent, and more preferably using a mixed solution of water and ethanol as a solvent. The mixed solution of water and ethanol is preferably an aqueous ethanol solution of 10% (v / v) to 90% (v / v), more preferably 30% (v / v) to 70% (v / v), still more preferably 40% (v / v) to 60% (v / v), and most preferably 45% (v / v) to 55% (v / v).

[0024] During extraction, the solvent can be used in an amount, for example, 1 to 40 times the dry weight of the extraction raw material, preferably 5 to 30 times, more preferably 10 to 30 times, still more preferably 10 to 25 times, and most preferably 15 to 20 times by weight.

[0025] The extraction temperature can be set, for example, at 0°C to 120°C, preferably 50°C to 100°C, more preferably 60°C to 90°C, still more preferably 70°C to 90°C, and most preferably 75°C to 85°C.

[0026] The extraction time can be set, for example, from 30 minutes to 12 hours, preferably from 1 hour to 10 hours, more preferably from 1 hour to 7 hours, still more preferably from 1 hour to 4 hours, and most preferably from 1 hour to 3 hours.

[0027] According to the above extraction conditions, extracts showing the desired effects of the present invention can be effectively prepared from marigold and basil.

[0028] In addition, the extract can be obtained by repeated extraction, for example, by repeating the process of separating and recovering the extract obtained after the first extraction from the residue, adding a new solvent to the residue, and obtaining the extract again by a second extraction. When performing repeated extraction, the number of repetitions can be, for example, 2 to 5 times, preferably 2 to 4 times, more preferably 2 to 3 times, and still more preferably 2 times.

[0029] In order to prepare the extract in a dry form, a drying process can be carried out after extraction. For example, a freeze-drying or spray-drying method after concentration under reduced pressure. The concentration under reduced pressure is preferably carried out at 50°C to 70°C under a pressure condition of 50 Torr to 100 Torr.

[0030] The marigold and basil extracts as the active ingredients of the present invention can be a mixture of a marigold extract and a basil extract, or can be an extract obtained from a mixture of marigold and basil.

[0031] When mixing the marigold extract and the basil extract, they can be mixed in a weight ratio ranging from, for example, 20:1 to 1:5, preferably 15:1 to 1:3, more preferably 12:1 to 1:2.5, and still more preferably 10:1 to 1:2.

[0032] When extracting the marigold and basil extracts from a mixture of marigold and basil, they can be extracts from a mixture in which marigold and basil are mixed in a weight ratio ranging from 20:1 to 1:5, preferably 15:1 to 1:3, more preferably 12:1 to 1:2.5, and still more preferably 10:1 to 1:2.

[0033] The composition of the present invention relates to a pharmaceutical composition for preventing or treating inflammatory diseases and a functional food composition for preventing or improving inflammatory diseases, and the composition of the present invention can prevent, treat, or improve inflammatory diseases through an effect on inflammation-related factors, for example, by inhibiting the increased expression of IL-6 and COX-2 caused by inducing an inflammatory response.

[0034] In the present invention, an inflammatory disease is a disease having inflammation as its main indication, and examples include inflammatory lung diseases, inflammatory liver diseases, inflammatory bowel diseases, autoinflammatory diseases, inflammatory central nervous system diseases, inflammatory skin diseases, and allergic inflammatory diseases, as well as arthritis, but the present invention is not limited thereto.

[0035] The composition of the present invention is preferably a pharmaceutical composition for preventing or treating arthritis or a functional food composition for preventing or improving arthritis.

[0036] More preferably, the composition of the present invention is a pharmaceutical composition for preventing or treating degenerative arthritis or a functional food composition for preventing or improving degenerative arthritis. In particular, the composition of the present invention can effectively prevent, treat, or improve degenerative arthritis by inhibiting the increased expression of MMP3 and MMP13 and the increased expression of IL-6 and COX-2.

[0037] The composition of the present invention can be orally or parenterally administered during clinical application and can be used in the form of a general pharmaceutical preparation. The pharmaceutical composition of the present invention may contain 0.1% to 100% by weight of calendula and basil extracts as active ingredients based on the total weight of the composition, but the present invention is not limited thereto.

[0038] In addition to calendula and basil extracts as active ingredients, the pharmaceutical composition of the present invention can be formulated into a conventional pharmaceutical dosage form by selecting one or two or more pharmaceutically acceptable carriers and one or two or more additives. The pharmaceutical composition of the present invention can be used alone or in combination not only with another pharmaceutically active compound but also with appropriate materials.

[0039] The pharmaceutical composition of the present invention can be formulated into various oral or parenteral dosage forms. For preparation, the pharmaceutical composition of the present invention can be formulated using diluents or excipients (such as common fillers, thickeners, binders, wetting agents, disintegrants and surfactants). Solid preparations for oral administration include tablets, pills, powders, granules and capsules, and such solid preparations are prepared by mixing at least one excipient (such as starch, calcium carbonate, sucrose, lactose or gelatin) with the composition. In addition to simple excipients, lubricants such as magnesium stearate, talc, etc. are also used. Liquid preparations for oral administration include suspensions, oral liquids, emulsions and syrups, and in addition to common simple diluents (such as water or liquid paraffin), various types of excipients such as wetting agents, sweeteners, flavoring agents and preservatives can be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations and suppositories. As non-aqueous solvents or suspensions, propylene glycol, polyethylene glycol, vegetable oils (such as olive oil) or injectable esters (such as ethyl oleate) can be used. As suppository bases, Witepsol, Tween 61, cocoa butter, laurin butter or glycerogelatin can be used.

[0040] In addition to the active ingredient, the pharmaceutical composition of the present invention may further contain: compounds including vitamins B, C, E or β-carotene, minerals such as Ca, Mg and Zn, phospholipids such as lecithin, or compounds such as maltol and amino acids.

[0041] In addition, in addition to the above components, known additives can also be used to enhance palatability, including natural flavors such as Japanese apricot, lemon, pineapple or herbal flavors; natural fruit juices; natural colorants such as chlorophyll and flavonoids; sweeteners such as fructose, honey, sugar alcohols and sugars; and sour agents such as citric acid and sodium citrate.

[0042] The subjects to which the pharmaceutical composition of the present invention can be administered can be vertebrates, preferably mammals, and more preferably humans.

[0043] The pharmaceutical composition of the present invention can be administered orally or parenterally. The pharmaceutical composition of the present invention can be administered by parenteral routes, including topical application to the skin or injection into the peritoneal cavity, rectum, vein, muscle, subcutaneous tissue, uterine cavity, epidural cavity or cerebral blood vessel, but preferably orally.

[0044] The dosage of the pharmaceutical composition of the present invention varies according to the patient's body weight, age, gender, health condition, diet, administration time, administration route, excretion rate, and the severity of the disease. For example, based on the amount of marigold and basil extracts, for an adult weighing 60 kg, the daily dosage range can be from 50 mg / day to 5,000 mg / day, and it can be administered once a day or divided into 2 to 6 doses per day. According to the animal experiment results, based on the amount of marigold and basil extracts, for an adult weighing 60 kg, the daily dosage can be 250 mg / day to 2,000 mg / day, preferably 500 mg / day to 2,000 mg / day, and more preferably 1,000 mg / day to 2,000 mg / day.

[0045] The pharmaceutical composition of the present invention may also contain one or more active ingredients that exhibit the same or similar functions.

[0046] The pharmaceutical composition of the present invention can be used alone or in combination with surgery, radiotherapy, hormone therapy, chemotherapy, or methods using biological response modifiers.

[0047] The food composition of the present invention can be, for example, various food products such as chewing gum, caramel products, candies, frozen desserts, and snacks; beverage products such as soft drinks, mineral water, and alcoholic beverages; and health functional foods containing vitamins, minerals, etc.

[0048] The marigold and basil extracts of the present invention can be used alone or in combination with other foods or food ingredients, and can be appropriately used according to conventional methods. The mixing amount of the active ingredient can be appropriately determined according to the purpose of use. Generally, in the production of food or beverage, the active ingredient can be added at 15% by weight or less, preferably 10% by weight or less, relative to the raw materials. However, in the case of long-term intake, the above amount can be the same as or lower than the above range, and the active ingredient can also be used in an amount greater than the above range.

[0049] The food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients. Natural carbohydrates may include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; and sugar alcohols such as xylitol, sorbitol, erythritol, etc. As sweeteners, natural sweeteners (such as miraculin and stevia extract) or synthetic sweeteners (such as saccharin and aspartame) can be used.

[0050] In addition to the above components, the food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerol, alcohols, and carbonating agents used in carbonated beverages. Additionally, the food composition of the present invention may contain the pulp used in the production of natural fruit juices, fruit juice beverages, and vegetable beverages. These components may be used alone or in combination.

[0051] Hereinafter, the present invention will be described in more detail with reference to the examples. However, these examples are provided only for illustrative purposes of the present invention, and thus the scope of the present invention should not be construed as being limited by these examples.

[0052] Preparation example

[0053] 1. Preparation of marigold extract

[0054] The above-ground parts of marigold are washed with water to remove surface impurities, and then cut and dried for subsequent use as extraction materials.

[0055] The prepared above-ground parts of marigold and a 50% (v / v) ethanol aqueous solution with a volume 20 times that of the above-ground parts of marigold are placed in a reflux extraction apparatus, and reflux extraction is carried out twice at 80 °C for 2 hours each time.

[0056] The extract produced by reflux extraction is filtered, concentrated, powdered, and used as an experimental sample.

[0057] 2. Preparation of basil extract

[0058] The above-ground parts of basil are washed with water to remove surface impurities, and then cut and dried for subsequent use as extraction materials.

[0059] The above-ground parts of basil and a 50% (v / v) ethanol aqueous solution with a volume 20 times that of the above-ground parts of basil are placed in a reflux extraction apparatus, and reflux extraction is carried out twice at 80 °C for 2 hours each time. Subsequently, the extract is filtered, concentrated, powdered, and used as an experimental sample.

[0060] Example 1

[0061] The marigold and basil extracts prepared in the preparation examples are mixed at a weight ratio of 1:1 and used as an experimental sample.

[0062] Examples 2 to 5

[0063] The marigold and basil extracts prepared in the preparation examples were mixed at a weight ratio of 4:1 (Example 2), 2:1 (Example 3), 1:0.1 (10:1; Example 4), and 1:2 (Example 5), and used as experimental samples.

[0064] Comparative example 1

[0065] The marigold extract prepared in the preparation examples was used alone as an experimental sample.

[0066] Comparative example 2

[0067] The basil extract prepared in the preparation examples was used alone as an experimental sample.

[0068] Experimental example 1

[0069] The aim was to study the effects of each sample on the expression of key inflammation-related factors (such as COX-2 and IL-6) and catabolic factors (such as MMP3 and MMP13) that cause degenerative arthritis.

[0070] Chondrocytes (primary chondrocytes) isolated from the knee joints of mice were used. Using ICR mouse P3, articular chondrocytes were isolated from mouse joints using collagenase and seeded at 2.88×10 5 cells per 35 mm culture dish and cultured in fetal bovine serum (FBS) / Dulbecco's modified Eagle's medium (DMEM). When the cell confluence reached approximately 70%, the cells were treated with IL-1β (1 ng / mL) and each sample at various concentrations and incubated for 24 hours.

[0071] Then, qRT-PCR was performed to examine whether the expression of MMP3, MMP13, IL-6, and COX-2, which increased in chondrocytes when inflammation (or degenerative arthritis) was induced by IL-1β treatment, was reduced by sample treatment.

[0072] Each of the prepared samples was dissolved in DMSO and used for the experiment.

[0073] The experimental results are shown in Figure 1 and Figure 2 .

[0074] As shown in Figure 1 , the composition of Example 1 (i.e., a mixture of marigold extract and basil extract at a weight ratio of 1:1) (in Figure 1In [description], it is represented as "marigold + basil") inhibits the increased expression of MMP3, MMP13, IL-6, and COX-2 caused by inflammation (or degenerative arthritis). In particular, compared with Comparative Example 1 (in Figure 1 represented as "marigold") and Comparative Example 2 (in Figure 1 represented as "basil"), the composition of Example 1 showed excellent inhibitory effects on the increased expression of all four factors, thus demonstrating the synergistic effect of the combination of marigold and basil.

[0075] In addition, as shown in Figure 2 , all the compositions of Examples 2 to 5 effectively inhibited the increased expression of MMP3, MMP13, IL-6, and COX-2 caused by inflammation (or degenerative arthritis) induced by IL-1β treatment, and showed inhibitory effects on the increased expression of all four factors similar to the levels observed in Example 1.

[0076] Experimental example 2

[0077] To study whether each sample is effective in improving osteoarthritis in an osteoarthritis animal model, mice, which are the most representative osteoarthritis animal model, were selected, and medial meniscus destabilization (DMM) surgery was performed to induce osteoarthritis.

[0078] For the DMM mouse model, DMM surgery was performed on 10-week-old male C57BL / 6 mice, which involved removing the fat pad and transecting the medial meniscus ligament to increase mechanical stress on the knee joint and induce cartilage degeneration and changes such as osteoarthritis. After DMM surgery, the mice were randomly grouped into phosphate-buffered saline (PBS), Comparative Example 1 (only marigold; represented as "Tagetes erecta" (200 mg / kg and 500 mg / kg)), Comparative Example 2 (only basil; represented as "Ocimum basilicum") (200 mg / kg and 500 mg / kg), and Example 4 (a combination of marigold and basil; represented as "WGA-M001") (50 mg / kg, 100 mg / kg, and 200 mg / kg), and each test material was orally administered to a group of 8 mice (n = 8 mice / group) at a predetermined dose daily for 10 weeks. In addition, to compare the efficacy of the test materials, as a positive control, health functional food ingredients that are currently widely used as health functional raw materials for joint health, such as Boswellia extract (represented as "Boswellia") and green-lipped mussel extract (represented as "Perna canaliculus"), were orally administered at a dose of 200 mg / kg daily in the same manner as the test materials.

[0079] Ten weeks after DMM surgery, cartilage samples from DMM-induced osteoarthritis mice were collected and fixed in 4% paraformaldehyde overnight, and then decalcified in 0.5 M EDTA for 2 weeks. Subsequently, the samples were embedded in paraffin and the paraffin blocks were cut into 5-μm-thick sections. The sections were stained with hematoxylin, safranin O, and Fast Green. Histopathological results were evaluated using the Osteoarthritis Research Society International (OARSI) grading system, osteophyte maturity grading, and measurement of subchondral bone plate (SBP) thickness. Representative safranin O-stained images were selected based on the area showing the most advanced lesions. For the experimental group, the OARSI score was evaluated blindly. The OARSI grading system consists of seven grades (0 to 6), where grade 0 indicates a joint without signs of osteoarthritis and grades 1 to 6 indicate different degrees of cartilage matrix abnormalities and the presence of cells in the superficial zone. Specifically, these grades reflect surface discontinuity, vertical cracks, erosion, delamination, and deformation, and the details are shown in Table 1 below.

[0080] [Table 1]

[0081]

[0082] As a result of daily oral administration of the marigold-basil complex to DMM-induced osteoarthritis mice to confirm its in vivo function, as shown by the safranin O staining results in Figure 3 , the marigold-basil complex (WGA-M001) exhibited a protective effect against the development of osteoarthritis. In particular, both marigold (Tagetes erecta) alone and basil (Ocimum basilicum) alone needed to be administered at a high dose (500 mg / kg) to prevent the development of osteoarthritis, but in the 100 mg / kg administration group, the marigold-basil complex (WGA-M001) showed similar osteoarthritis inhibitory and protective effects to the high-dose group. Additionally, in fact, the results of OARSI grading, SBP thickness, and bone tissue maturity measurement were consistent with the results of articular cartilage tissue analysis observed by safranin O staining ( Figures 4 to 6 ). This shows that the articular protective effect of the marigold-basil complex is at least 5 times that of other extracts, and the combined use of marigold and basil has a synergistic effect, thus demonstrating a cartilage restoration effect even at a lower concentration than when they are used alone.

[0083] Refer to Figure 3In the PBS (negative control) group, compared with the normal group (Sham), the infiltration of inflammatory cells into the joints and cartilage was significantly increased in mice with DMM-induced osteoarthritis. On the other hand, in the groups treated with Tagetes erecta extract, Ocimum basilicum extract, and Tagetes erecta-Ocimum basilicum complex (WGA-M001), the inflammatory response was alleviated, and particularly, a significant alleviation of the inflammatory response was confirmed in the group treated with the Tagetes erecta-Ocimum basilicum complex (WGA-M001).

[0084] In addition, the subchondral bone plate (SBP) thickness of mice with osteoarthritis induced by DMM surgery was measured and found to be greatly increased in the negative control (PBS) without any experimental material administration, but was found to be greatly decreased in the group administered with the Tagetes erecta-Ocimum basilicum complex (WGA-M001). This result can show that the Tagetes erecta-Ocimum basilicum complex has the effect of improving osteoarthritis. In addition, compared with the Boswellia extract and Perna canaliculus extract, which are currently used as health functional raw materials for joint health care, the Tagetes erecta-Ocimum basilicum complex (WGA-M001) showed a greater inhibitory effect on osteoarthritis.

Claims

1. A pharmaceutical composition for preventing or treating inflammatory diseases, said pharmaceutical composition comprising calendula and basil extracts as active ingredients.

2. The pharmaceutical composition according to claim 1, wherein the extract is a 10% (v / v) to 90% (v / v) ethanol extract.

3. The pharmaceutical composition according to claim 1, wherein the inflammatory disease is arthritis.

4. The pharmaceutical composition according to claim 3, wherein the arthritis is degenerative arthritis.

5. The pharmaceutical composition according to claim 1, wherein the extract is an extract of a mixture of calendula and basil mixed in a weight ratio ranging from 15:1 to 1:

3.

6. A functional food composition for preventing or improving inflammatory diseases, said functional food composition comprising calendula and basil extracts as active ingredients.

7. The functional food composition according to claim 6, wherein the extract is a 10% (v / v) to 90% (v / v) ethanol extract.

8. The functional food composition according to claim 6, wherein the inflammatory disease is arthritis.

9. The functional food composition according to claim 8, wherein the arthritis is degenerative arthritis.

10. The functional food composition according to claim 6, wherein the extract is an extract of a mixture of calendula and basil mixed in a weight ratio ranging from 15:1 to 1:3.

Images