Composition for preventing, treating or ameliorating inflammatory diseases

Abstract

The pharmaceutical composition for preventing or treating inflammatory diseases of the present invention contains marigold and basil extracts as active ingredients. Also, the functional food composition for preventing or ameliorating 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 ameliorating inflammatory diseases. Specifically, the present invention relates to a composition that utilizes plant materials and can effectively prevent, treat, or ameliorate inflammatory diseases by regulating the expression of inflammatory disease-related factors. [Background technology]

[0002] The inflammatory response is a biological response to any stimulus, such as tissue damage or microbial infection, and is essential for maintaining the normal function of the body. However, if this inflammatory response occurs excessively, the tissues and organs at the site of inflammation are unable to maintain their normal functions, resulting in pain and fever. Diseases caused by such excessive inflammatory responses are called inflammatory diseases. Although much research has been conducted on anti-inflammatory drugs to treat such inflammatory diseases, there is a continuing demand for safer and more effective anti-inflammatory drugs.

[0003] Arthritis is a representative inflammatory disease. Arthritis is a disease in which damage and inflammation occur due to various causes in the joints, which are the joints between bones. Many people suffer from arthritis, which can particularly impair patients' mobility and significantly impact their social activities. Among arthritis types, degenerative arthritis is a progressive, irreversible degenerative disease that reduces quality of life. It is the most common degenerative disease, affecting eight out of ten elderly people. Osteoarthritis, in particular, causes pain and impairments in daily activities and mobility, reducing quality of life and consuming astronomical social costs for treatment. However, there are currently no therapeutic agents for this condition.

[0004] Osteoarthritis is a disease that causes the degeneration of articular cartilage tissue and structural changes in the subchondral bone. While the pathological causes are still unknown, the main causes include cartilage wear due to repeated use during the aging process, genetics, damage to cartilage tissue due to impact, pressure due to obesity, and weakened muscles around the knee. In particular, congenital factors such as aging and genetics, and mechanical factors such as obesity, injury, and weakened muscles activate biochemical pathways that induce osteoarthritis.

[0005] In order to develop pharmaceutical compositions and functional food compositions that can effectively prevent, treat or improve the above-mentioned inflammatory diseases, particularly arthritis, especially osteoarthritis, the present inventors searched for substances from plant materials that are effective against these diseases, and found that a combination of marigold and basil is highly effective in treating these diseases, leading to the completion of the present invention. Summary of the Invention [Problem to be solved by the invention]

[0006] The object of the present invention is to provide a pharmaceutical composition and a functional food composition that utilize plant materials to effectively prevent, treat or improve inflammatory diseases, particularly arthritis, especially osteoarthritis. [Means for solving the problem]

[0007] In one aspect, the present invention provides a pharmaceutical composition for preventing or treating inflammatory diseases, comprising marigold and basil extracts as active ingredients.

[0008] In another aspect, the present invention provides a functional food composition for preventing or ameliorating inflammatory diseases, which contains marigold and basil extracts as active ingredients. [Effects of the Invention]

[0009] The composition of the present invention utilizes plant materials and can effectively prevent, treat, or improve inflammatory diseases, particularly arthritis, especially osteoarthritis, by suppressing the increased expression of inflammatory response-induced factors such as MMP3, MMP13, IL-6, and COX-2. In particular, the composition of the present invention can exert a synergistic effect by combining two plant materials. [Brief explanation of the drawings]

[0010] [Figure 1] Chondrocytes isolated from mouse knee joints were treated with IL-1β (1 ng / ml) to induce osteoarthritis, and the expression levels of MMP3, MMP13, IL-6, and COX-2 were examined by qRT-PCR in mice treated with a 1:1 mixture of marigold extract and basil extract (marigold + basil), marigold extract alone (marigold), and basil extract alone (basil). *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, ns=not significant. [Figure 2] Chondrocytes isolated from mouse knee joints were treated with IL-1β (1 ng / ml) to induce osteoarthritis, and the expression levels of MMP3, MMP13, IL-6, and COX-2 were examined by qRT-PCR when 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, ns=not significant. [Figure 3]The results show the therapeutic effects of various concentrations of experimental substances administered to mice with osteoarthritis induced by medial meniscal cartilage damage (DMM) surgery. Normal mice (Sham) and mice with osteoarthritis induced by DMM were administered various concentrations of the experimental substances for 10 weeks, and then autopsied. The photographs show the joints, cartilage, and synovium stained with sapranin O. The negative control group was administered PBS (phosphate-buffered saline); the group administered WGA-M001, a complex of marigold extract and basil extract; the group administered Tagetes erecta (marigold extract alone); the group administered Ocimum basilicum (basil extract alone); the positive control group administered Boswellia (boswellia extract, traditionally used as a functional ingredient for joint health); and the positive control group administered Perna canaliculus (green-lipped mussel extract, traditionally used as a functional ingredient for joint health). [Figure 4] This graph shows the grade of osteoarthritis according to the OARSI assessment method for mice with osteoarthritis induced by DMM. Sham, normal mice; DMM, mice with osteoarthritis induced by DMM; PBS, negative control group administered with phosphate-buffered saline; WGA-M001, a group administered with a complex of marigold extract and basil extract; Tagetes erecta, a group administered with marigold extract alone; Ocimum basilicum, a group administered with basil extract alone; Boswellia, a positive control group administered with Boswellia extract; and Perna canaliculus, a positive control group administered with green-lipped mussel extract. Values ​​are expressed as mean ± SD and evaluated using the Kruskal-Wallis test and Mann-Whitney U test. Significant differences from the negative control group (PBS) were indicated as *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, and ns=not significant. [Figure 5]This graph shows the subchondral bone plate thickness measured by the administration of various concentrations of experimental substances in mice with osteoarthritis induced by DMM. Sham, normal mice; DMM, mice with osteoarthritis induced by DMM; PBS, negative control group administered with phosphate-buffered saline; WGA-M001, a group administered with a complex of marigold extract and basil extract; Tagetes erecta, a group administered with marigold extract alone; Ocimum basilicum, a group administered with basil extract alone; Boswellia, positive control group administered with Boswellia extract; and Perna canaliculus, positive control group administered with green-lipped mussel extract. Values ​​are expressed as mean ± SD and evaluated using the Kruskal-Wallis test and Mann-Whitney U test. Significant differences from the negative control group (PBS) were indicated as *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, and ns=not significant. [Figure 6] This graph shows the degree of osteophyte maturation (degree of formation) in mice with osteoarthritis induced by DMM (Drug-Method-Induced Mice). Sham: normal mice; DMM: mice with osteoarthritis induced by DMM; PBS: negative control group administered with phosphate-buffered saline; WGA-M001: group administered with a complex of marigold and basil extracts; Tagetes erecta: group administered with marigold extract alone; Ocimum basilicum: group administered with basil extract alone; Boswellia: positive control group administered with Boswellia extract; and Perna canaliculus: positive control group administered with green-lipped mussel extract. Values ​​are expressed as mean ± SD and evaluated using the Kruskal-Wallis test and Mann-Whitney U test. Significant differences from the negative control (PBS) were indicated as *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, and ns=not significant. DETAILED DESCRIPTION OF THE INVENTION

[0011] The composition of the present invention is characterized by containing marigold and basil extracts as active ingredients.

[0012] Marigold is an annual plant of the genus Tagetes in the Asteraceae family, and is primarily used as tea or as a food flavoring or coloring agent. In the present invention, a marigold selected from the group consisting of African marigold (Tagetes erecta), French marigold (T. patula), wild marigold (T. minuta), and Mexican marigold (T. lucida) may be used, with African marigold being preferred. Furthermore, in the present invention, the aerial parts of such marigolds may be used as the extraction raw material.

[0013] Basil is an annual plant of the Lamiaceae family, commonly used in Italian and French cuisine. In the present invention, a basil selected from the group consisting of sweet basil (Ocimum basilicum), holy basil (O. tenuiflorum), American basil (O. americanum), Amazonian basil (O. campechianum), and African basil (O. gratissimum) may be used, with sweet basil being preferred. Furthermore, the above-ground parts of such basil may be used as the extraction material in the present invention.

[0014] In the present invention, the term "extract" may refer to a liquid component obtained by immersing an extraction material in a solvent and then extracting for a certain period of time, for example, at room temperature or under heated conditions, or a solid component obtained by removing the solvent from the liquid component. Furthermore, the term "extract" may be interpreted as encompassing not only the above-mentioned product, but also dilutions of the above-mentioned product, concentrates thereof, and crude and purified products thereof.

[0015] The extract of the present invention can be prepared using marigold and basil as the extraction raw material and extraction methods commonly used in this field. For example, extraction can be performed using a polar or nonpolar solvent. The polar solvent may be selected from the group consisting of C1-C4 lower alcohols and water, and the nonpolar solvent may be selected from the group consisting of chloroform, petroleum ether, ether, hexane, toluene, ethyl acetate, dichloromethane, and the like. To achieve the desired effects of the present invention, the extract of the present invention is preferably extracted using water, ethanol, or a mixture thereof as the solvent, more preferably a mixture of water and ethanol. The water and ethanol mixture is preferably a 10-90% (v / v) aqueous ethanol solution, more preferably a 30-70% (v / v) aqueous ethanol solution, more preferably a 40-60% (v / v) aqueous ethanol solution, and even more preferably a 45-55% (v / v) aqueous ethanol solution.

[0016] During extraction, for example, 1 to 40 times the weight of the solvent relative to the dry weight of the extraction raw material may be used, and preferably 5 to 30 times, more preferably 10 to 30 times, more preferably 10 to 25 times, and more preferably 15 to 20 times the weight of the solvent relative to the dry weight of the extraction raw material.

[0017] The extraction temperature may be set, for example, at 0 to 120°C, preferably at 50 to 100°C, more preferably at 60 to 90°C, more preferably at 70 to 90°C, and more preferably at 75 to 85°C.

[0018] The extraction time may be set, for example, from 30 minutes to 12 hours, and is preferably set to from 1 to 10 hours, more preferably from 1 to 7 hours, more preferably from 1 to 4 hours, and more preferably from 1 to 3 hours.

[0019] According to the above extraction conditions, an extract exhibiting the desired effects of the present invention can be efficiently produced from marigold and basil.

[0020] Alternatively, the extraction can be repeated, for example, by separating and recovering the extract from the residue after the first extraction, adding a new solvent to the residue, and performing a second extraction to obtain the extract again. When the extraction is repeated, the number of repetitions may be, for example, 2 to 5 times, preferably 2 to 4 times, more preferably 2 to 3 times, and more preferably 2 times.

[0021] To prepare a dried extract, a drying process may be carried out after extraction. For example, the extract may be concentrated under reduced pressure followed by freeze-drying or spray-drying. The reduced pressure concentration is preferably carried out at a temperature of 50 to 70°C and a pressure of 50 to 100 torr.

[0022] The marigold and basil extract of the present invention may be a mixture of marigold extract and basil extract, or may be extracted from a mixture of marigold and basil.

[0023] When marigold extract and basil extract are mixed, for example, the marigold extract and basil extract may be mixed in a weight ratio of 20:1 to 1:5, preferably in a weight ratio of 15:1 to 1:3, more preferably in a weight ratio of 12:1 to 1:2.5, and even more preferably in a weight ratio of 10:1 to 1:2.

[0024] When extracted from a mixture of marigold and basil, it may be, for example, an extract of a mixture of marigold and basil in a weight ratio of 20:1 to 1:5, preferably an extract of a mixture of marigold and basil in a weight ratio of 15:1 to 1:3, more preferably a 12:1 to 1:2.5, and more preferably a 10:1 to 1:2.

[0025] 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 ameliorating inflammatory diseases, and the composition of the present invention can prevent, treat, or ameliorate inflammatory diseases through its effect on inflammation-related factors, for example, by suppressing the increased expression of IL-6 and COX-2 due to the induction of an inflammatory response.

[0026] In the present invention, inflammatory diseases refer to diseases whose main lesion is inflammation, and include, but are not limited to, arthritis, inflammatory lung diseases, inflammatory liver diseases, inflammatory bowel diseases, autoinflammatory diseases, inflammatory central nervous system diseases, inflammatory skin diseases, and allergic inflammatory diseases.

[0027] The composition of the present invention is preferably a pharmaceutical composition for preventing or treating arthritis or a functional food composition for preventing or ameliorating arthritis.

[0028] More preferably, the composition of the present invention is a pharmaceutical composition for preventing or treating osteoarthritis or a functional food composition for preventing or ameliorating osteoarthritis. In particular, the composition of the present invention can effectively prevent, treat, or ameliorate osteoarthritis through its inhibitory effect on the increased expression of IL-6 and COX-2, as well as MMP3 and MMP13.

[0029] The composition of the present invention can be administered orally or parenterally in clinical administration and may be used in the form of a common pharmaceutical formulation. The pharmaceutical composition of the present invention may contain, for example, 0.1 to 100% by weight of marigold and basil extract as an active ingredient based on the total weight of the composition, but is not limited thereto.

[0030] The pharmaceutical composition of the present invention may be formulated into a conventional pharmaceutical preparation by selecting one or more pharmaceutically acceptable carriers and one or more additives in addition to the active ingredients, marigold and basil extract. The pharmaceutical composition of the present invention may be used alone or in combination with other pharmaceutically active compounds, as well as in combination with other appropriate substances.

[0031] The pharmaceutical compositions of the present invention may be in various oral or parenteral dosage forms. When formulated, they may be prepared using commonly used diluents or excipients, such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid formulations for oral administration include tablets, pills, powders, granules, capsules, and the like. These solid formulations may be prepared by mixing one or more compounds with at least one or more excipients, such as starch, calcium carbonate, sucrose or lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid formulations for oral administration include suspensions, oral solutions, emulsions, syrups, and the like. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients, such as wetting agents, sweeteners, flavoring agents, and preservatives, may also be included. Preparations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspension solvents may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases may include witepsol, macrogol, Tween 61, cocoa butter, laurin butter, and glycerogelatin.

[0032] In addition to the active ingredient, the pharmaceutical composition of the present invention may contain auxiliary ingredients such as vitamins B, C, and E, β-carotene, mineral-containing compounds such as Ca, Mg, and Zn, phospholipids such as lecithin, or compounds such as maltol and amino acids.

[0033] In addition to the above ingredients, known additives may be used to enhance the taste, such as natural flavors such as plum, lemon, pineapple, or herb flavors, natural fruit juices, natural pigments such as chlorphyllin and flavonoids, and sweeteners such as fructose, honey, sugar alcohols, and sugars, as well as acidulants such as citric acid and sodium citrate.

[0034] The individuals to which the pharmaceutical compositions of the present invention can be applied are vertebrates, preferably mammals, and more preferably humans.

[0035] The pharmaceutical composition of the present invention may be administered orally or parenterally. When administered parenterally, topical application to the skin or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebrovascular injection may be used, and oral administration is preferred.

[0036] The dosage of the pharmaceutical composition of the present invention varies depending on the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of disease. For example, the daily dosage may be 50-5,000 mg / day for a 60 kg adult based on the amount of marigold and basil extract, and may be administered once or in 2-6 divided doses per day. Based on the results of animal experiments, the daily dosage is preferably 250-2,000 mg / day for a 60 kg adult based on the amount of marigold and basil extract, more preferably 500-2,000 mg / day for a 60 kg adult based on the amount of marigold and basil extract, and even more preferably 1,000-2,000 mg / day for a 60 kg adult based on the amount of marigold and basil extract.

[0037] The pharmaceutical composition of the present invention may further contain one or more active ingredients exhibiting the same or similar function.

[0038] The pharmaceutical compositions of the present invention may be used alone or in combination with methods involving surgery, radiation therapy, hormone therapy, chemotherapy and the use of biological response modifiers.

[0039] The food composition of the present invention may be, for example, various foods such as chewing gum, caramel products, candies, ice cream, and confectioneries; beverage products such as soft drinks, mineral water, and alcoholic beverages; and health functional foods containing vitamins and minerals.

[0040] The marigold and basil extract of the present invention may be used alone or in combination with other foods or food ingredients, and may be used appropriately in a conventional manner. The amount of active ingredient to be added may be appropriately determined depending on the intended use. Generally, when producing a food or beverage, the active ingredient of the present invention may be added in an amount of 15% by weight or less, preferably 10% by weight or less, based on the raw materials. However, for long-term intake, the amount may be less than the above range, and the active ingredient may be used in an amount greater than the above range.

[0041] The food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients. Natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, or sugar alcohols such as xylitol, sorbitol, and erythritol. Natural sweeteners such as thaumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame may also be used as sweeteners.

[0042] In addition, the food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectinic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, etc. In addition, the food composition of the present invention may contain fruit pulp for the production of natural fruit juices, fruit juice beverages, and vegetable beverages. These ingredients may be used alone or in combination.

[0043] The present invention will be described in more detail below through examples, which are merely for the purpose of illustrating the present invention and are not to be construed as limiting the scope of the present invention.

[0044] (Manufacturing example) (1. Production of Marigold Extract) The whole marigold plant (above ground) was washed with water to remove foreign matter from the surface, cut, dried and used as the extraction material.

[0045] The prepared marigold whole plant and 20 times its weight of 50% (v / v) ethanol aqueous solution were placed in a reflux extraction apparatus and reflux extracted at 80°C for 2 hours twice.

[0046] The extract produced by reflux extraction was filtered, concentrated and powdered, which was then used as the experimental sample.

[0047] (2. Production of Basil Extract) The whole basil plant (above ground) was washed with water to remove foreign matter from the surface, cut, dried and used as the extraction material.

[0048] Whole basil and 20 times its weight of 50% (v / v) ethanol aqueous solution were placed in a reflux extraction apparatus and reflux extracted twice at 80°C for 2 hours. The extract was then filtered, concentrated, and powdered, and used as the experimental sample.

[0049] Example 1 The marigold extract and basil extract prepared in the above Preparation Example were mixed in a weight ratio of 1:1 and used as the experimental sample.

[0050] Examples 2 to 5 The marigold extract and basil extract prepared in the above Preparation Examples were mixed in a weight ratio of 4:1 (Example 2), 2:1 (Example 3), 1:0.1 (10:1; Example 4), or 1:2 (Example 5), and the mixture was used as the experimental sample.

[0051] (Comparative Example 1) The marigold extract prepared in the above Preparation Example was used alone as an experimental sample.

[0052] (Comparative Example 2) The basil extract prepared in the above Preparation Example was used alone as the experimental sample.

[0053] (Experimental Example 1) The effects of each sample on the expression of COX-2 and IL-6, which are representative inflammation-related factors, and MMP3 and MMP13, which are catabolic factors that induce osteoarthritis, were examined.

[0054] Primary chondrocytes isolated from the knee joints of mice were used. ICR mice P3 were used. Chondrocytes were isolated from the joints of mice using collagenase. Articular chondrocytes were cultured in FBS (Fetal Bovine Serum) / Dulbecco's Modified Eagle Medium (DMEM) medium at 2.88 × 10 cells per 35 mm dish. 5 When the cells reached about 70% confluence, they were treated with IL-1β (1 ng / ml) and each sample at different concentrations and cultured for 24 hours.

[0055] When inflammation (or osteoarthritis) is induced by IL-1β treatment, qRT-PCR was performed to examine whether the expression of MMP3, MMP13, IL-6, and COX-2, which increases in chondrocytes, is reduced by sample treatment.

[0056] Each sample was dissolved in DMSO and used in the experiment.

[0057] The experimental results are shown in Figures 1 and 2.

[0058] As shown in FIG. 1, the composition of Example 1, i.e., a mixture of marigold extract and basil extract in a 1:1 weight ratio (shown as "Marigold + Basil" in FIG. 1), inhibited the increased expression of MMP3, MMP13, IL-6, and COX-2 caused by inflammation (or osteoarthritis) induction (IL-1β treatment). In particular, when compared with Comparative Example 1 (shown as "Marigold" in FIG. 1) and Comparative Example 2 (shown as "Basil" in FIG. 1), the composition of Example 1 exhibited a superior inhibitory effect on the increased expression of all four factors, indicating a synergistic effect due to the combination of marigold and basil.

[0059] Furthermore, as shown in Figure 2, all of the compositions of Examples 2 to 5 were shown to effectively suppress the increased expression of MMP3, MMP13, IL-6, and COX-2 caused by inflammation (or osteoarthritis) induction (treatment with IL-1β), and all four factors showed the same level of inhibitory effect on increased expression as in Example 1.

[0060] (Experimental Example 2) To examine the effect of the sample on improving osteoarthritis in an animal model of osteoarthritis, we selected a mouse model, which is a very representative animal model for osteoarthritis, in which osteoarthritis was induced by performing a medial meniscus cartilage injury (DMM) surgery on the mouse.

[0061] In the DMM mouse model, 10-week-old male C57BL / 6 mice underwent DMM surgery, which involved removal of the fat pad and incision of the medial meniscus-tibial ligament to increase mechanical stress in the knee joint, inducing cartilage degradation and osteoarthritis-like changes. After DMM surgery, mice were randomly assigned to one of the following groups: phosphate-buffered saline (PBS), comparative example 1 (marigold alone; labeled "Tagetes erecta") (200 and 500 mg / kg), comparative example 2 (basil alone; labeled "Ocimum basilicum") (200 and 500 mg / kg), or example 4 (marigold and basil combination; labeled "WGA-M001") (50, 100, and 200 mg / kg). Eight mice per group (n=8) were orally administered the prescribed doses daily for 10 weeks. In order to compare the efficacy of the experimental substances separately, boswellia extract (labeled "Boswellia") and green-lipped mussel extract (labeled "Perna canaliculus"), which are currently widely used functional food ingredients for joint health, were used as positive controls and were orally administered daily at a dose of 200 mg / kg, just like the experimental substances.

[0062] Ten weeks after DMM surgery, cartilage samples from DMM-induced osteoarthritic mice were harvested, fixed overnight in 4% paraformaldehyde, and decalcified in 0.5 M EDTA for 2 weeks. The samples were then embedded in paraffin, and the paraffin blocks were cut into 5 μm-thick sections. The sections were stained with hematoxylin, sapranin O, and fast green. Histopathological results were scored using the Osteoarthritis Research Society International (OARSI) grade, osteophyte maturation grade, and subchondral bone thickness measurement. Representative sapranin O-stained images were selected from the most advanced lesions. The OARSI score was assessed for each experimental group using a blinded observational method. The OARSI scoring system consists of seven grades (0–6), with grade 0 representing a joint without signs of osteoarthritis and grades 1–6 representing various levels of cartilage matrix abnormalities and the presence of superficial cells. Specifically, these include surface discontinuities, vertical cracks, erosion, delamination and deformation, and are detailed in Table 1 below.

[0063] [Table 1]

[0064] To verify the in vivo function of the marigold and basil complex, the corresponding samples were orally administered daily to mice with DMM-induced osteoarthritic disease. As can be seen from the safranin O staining results in Figure 3, the marigold and basil complex (WGA-M001) demonstrated a protective effect against the development of osteoarthritis. While marigold (Tagetes erecta) and basil (Ocimum basilicum) alone require high doses (500 mg / kg) to block the development of osteoarthritis, the marigold and basil complex (WGA-M001) at 100 mg / kg demonstrated similar inhibitory and protective effects against the development of osteoarthritis as the high-dose group. Furthermore, the OARSI grade, SBP thickness, and bone maturity measurements were consistent with the results of autopsy analysis of articular cartilage tissue observed with safranin O staining (Figures 4-6). This indicates that the joint-protecting effect of the marigold and basil complex is at least five times stronger than that of other extracts, and that there is a synergistic effect when using a combination of marigold and basil, with a cartilage-restoring effect even at lower concentrations than when using either extract alone.

[0065] As shown in Figure 3 (PBS, negative control group), mice with osteoarthritis induced by DMM surgery showed significantly increased infiltration of inflammatory cells into the joints and cartilage compared to the normal (Sham) group. Meanwhile, the inflammatory response was alleviated in the groups treated with marigold extract (Tagetes erecta), basil extract (Ocimum basilicum), and marigold and basil complex (WGA-M001), with the marigold and basil complex (WGA-M001) showing particularly significant alleviation of the inflammatory response.

[0066] In addition, the thickness of the subchondral bone plate (SBP) was measured in mice with osteoarthritis induced by the DMM surgery. The negative control group (PBS), which did not receive any experimental substances, showed a significant increase in SBP thickness, while the group administered the marigold and basil complex (WGA-M001) showed a significant decrease in SBP thickness. These results demonstrate that the marigold and basil complex has an effect in improving osteoarthritis. Furthermore, the marigold and basil complex (WGA-M001) showed greater effectiveness in suppressing osteoarthritis than boswellia extract and green-lipped mussel extract (Perna canaliculus), which are currently used as functional ingredients for joint health.

[0067] (Addendum) (Appendix 1) A pharmaceutical composition for preventing or treating inflammatory diseases, comprising marigold and basil extracts as active ingredients.

[0068] (Appendix 2) 2. The pharmaceutical composition for preventing or treating an inflammatory disease according to claim 1, wherein the extract is a 10 to 90% (v / v) ethanol extract.

[0069] (Appendix 3) 2. The pharmaceutical composition for preventing or treating an inflammatory disease according to claim 1, wherein the inflammatory disease is arthritis.

[0070] (Appendix 4) The pharmaceutical composition for preventing or treating an inflammatory disease according to Appendix 3, wherein the arthritis is osteoarthritis.

[0071] (Appendix 5) 2. The pharmaceutical composition for preventing or treating an inflammatory disease according to claim 1, wherein the extract is an extract of a mixture of marigold and basil in a weight ratio of 15:1 to 1:3.

[0072] (Appendix 6) A functional food composition for preventing or improving inflammatory diseases, comprising marigold and basil extracts as active ingredients.

[0073] (Appendix 7) 7. The functional food composition for preventing or ameliorating an inflammatory disease according to Appendix 6, wherein the extract is a 10 to 90% (v / v) ethanol extract.

[0074] (Appendix 8) 7. The functional food composition for preventing or ameliorating an inflammatory disease according to claim 6, wherein the inflammatory disease is arthritis.

[0075] (Appendix 9) The functional food composition for preventing or ameliorating an inflammatory disease according to Appendix 8, wherein the arthritis is osteoarthritis.

[0076] (Appendix 10) 7. The functional food composition for preventing or ameliorating an inflammatory disease according to Appendix 6, wherein the extract is an extract of a mixture of marigold and basil in a weight ratio of 15:1 to 1:3.

Claims

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

2. 2. The pharmaceutical composition for preventing or treating an inflammatory disease according to claim 1, wherein the extract is a 10 to 90% (v / v) ethanol extract.

3. The pharmaceutical composition for preventing or treating an inflammatory disease according to claim 1, wherein the inflammatory disease is arthritis.

4. The pharmaceutical composition for preventing or treating an inflammatory disease according to claim 3, wherein the arthritis is osteoarthritis.

5. 2. The pharmaceutical composition for preventing or treating an inflammatory disease according to claim 1, wherein the extract is an extract of a mixture of marigold and basil in a weight ratio of 15:1 to 1:

3.

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

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

8. The functional food composition for preventing or ameliorating an inflammatory disease according to claim 6, wherein the inflammatory disease is arthritis.

9. The functional food composition for preventing or ameliorating an inflammatory disease according to claim 8, wherein the arthritis is osteoarthritis.

10. 7. The functional food composition for preventing or improving inflammatory diseases according to claim 6, wherein the extract is an extract of a mixture of marigold and basil in a weight ratio of 15:1 to 1:3.

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