Pharmaceutical composition for preventing or treating degenerative osteoarthritis

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating degenerative osteoarthritis, containing a pyrano[2,3-f]chromene derivative compound, a pharmaceutically acceptable salt thereof, or a solvate thereof.

Description

Pharmaceutical composition for the prevention or treatment of degenerative osteoarthritis

[0001] The present invention relates to a pharmaceutical composition for preventing or treating degenerative arthritis.

[0002] Osteoarthritis is a disease in which the articular cartilage covering the joint surfaces of bones wears away, exposing the underlying bone. This inflammation causes inflammation in the synovial membrane surrounding the joint, resulting in pain and deformity. Commonly referred to as degenerative arthritis, it is the most common joint disease. Osteoarthritis is caused by damage to the soft cartilage that acts as a cushion between bones. Causes include genetic factors, obesity, joint trauma, and cartilage damage due to inflammation. Chronic joint disease, starting in childhood, can lead to osteoarthritis, which can develop at a relatively young age. Spinal osteoarthritis can cause lower back pain. In severe cases, it can cause tingling or numbness, sometimes mistaken for a herniated disc. Hip osteoarthritis can cause pain or limited range of motion, making walking awkward. Osteoarthritis is also common in the finger joints, particularly in middle-aged women. Finger osteoarthritis can cause pain and a gradual thickening of the finger joints.

[0003] Treatment options for osteoarthritis include medication, physical therapy, lifestyle modifications, and surgery. Most medications reduce pain and swelling and slow the progression of the disease, but they are not fundamental cures. The most commonly used medications are nonsteroidal anti-inflammatory drugs (NSAIDs) and steroid injections. While NSAIDs offer excellent anti-inflammatory and analgesic effects, long-term use can lead to gastrointestinal side effects or cardiovascular problems such as heart attack, stroke, and myocardial infarction. Furthermore, steroid injections have only temporary effects, and frequent use can actually damage cartilage.

[0004] The pain experienced by osteoarthritis patients is nociceptive pain, a signal that cartilage tissue needs protection and recovery from damage. Nociceptive pain typically resolves quickly when the threatening stimulus is removed or tissues heal. However, in osteoarthritis patients, pain persists even after the external stimulus is removed. This is because osteoarthritis is an endogenous disease, originating from within the body itself, rather than an external stimulus. Degenerative diseases are closely linked to aging, and cell metabolism declines with age, leading to apoptosis. Mitochondrial dysfunction and decreased metabolism in chondrocytes of osteoarthritis patients gradually induce apoptosis, which the body perceives as a warning signal, resulting in persistent pain. Therefore, improving cartilage cell metabolism and inhibiting apoptosis is a novel osteoarthritis treatment that not only alleviates pain but also fundamentally addresses the endogenous cause.

[0005]

[0006] The purpose of the present invention is to provide a pharmaceutical composition for preventing or treating degenerative arthritis.

[0007] However, the problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by those skilled in the art from the following film.

[0008]

[0009] One embodiment of the present invention provides a pharmaceutical composition for preventing or treating degenerative osteoarthritis, comprising a pyrano[2,3-f]chromene derivative compound of the following chemical formula 1, a pharmaceutically acceptable salt thereof, or a solvate thereof:

[0010] [Chemical Formula 1]

[0011]

[0012] In the above chemical formula 1, R1 is a hydrogen atom; a substituted or unsubstituted, straight-chain or branched C1-C6 alkyl group; a halogen atom; a substituted or unsubstituted, straight-chain or branched C1-C6 alkoxy group; or a substituted or unsubstituted, straight-chain or branched C1-C4 thioalkyl group; and R2 is a hydrogen atom, methyl, ethyl, methoxy, or ethoxy, and R3 and R4 are each independently a hydrogen atom or a C1-C2 alkyl group, and in the case of the substituted alkyl, substituted alkoxy, and substituted thioalkyl, the substituent is a straight-chain or branched C1-C5 alkyl group, a halogen atom, a straight-chain or branched C1-C5 alkoxy group, or a straight-chain or branched C1-C3 thioalkyl group.

[0013]

[0014] A pharmaceutical composition comprising a pyrano[2,3-f]chromene derivative compound, a pharmaceutically acceptable salt thereof, or a solvate thereof according to one embodiment of the present invention has excellent preventive and therapeutic efficacy against degenerative arthritis.

[0015] The effects of the present invention are not limited to the effects described above, and effects not mentioned will be clearly understood by those skilled in the art from the present specification and the attached drawings.

[0016]

[0017] Figure 1 shows the results of measuring pain levels in a MIA-OA rat model according to Experimental Example 1.

[0018] Figure 2 shows the results of measuring body weight load in an MIA-OA rat model according to Experimental Example 2.

[0019] Figure 3 shows the results of knee joint tissue pathology of the MIA-OA rat model according to Experimental Example 3.

[0020] Figure 4 shows the results of measuring OARSI and Mankin scores through tissue pathology results according to Experimental Example 3.

[0021] Figure 5 shows the results of quantifying IL-6 changes in knee joint tissue through tissue immunochemical analysis according to Experimental Example 4.

[0022] Figure 6 shows the results of quantifying changes in MCP-1 in knee joint tissue through tissue immunochemical analysis according to Experimental Example 4.

[0023] Figure 7 shows the results of quantifying changes in MMP3 in knee joint tissue through tissue immunochemical analysis according to Experimental Example 4.

[0024] Figure 8 shows the results of quantifying changes in RIPK3 in knee joint tissue through tissue immunochemical analysis according to Experimental Example 4.

[0025]

[0026] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by those skilled in the art in the relevant fields of the present invention. Furthermore, while preferred methods and samples are described herein, similar or equivalent methods are also included within the scope of the present invention.

[0027]

[0028] One embodiment of the present invention provides a pharmaceutical composition for preventing or treating degenerative osteoarthritis, comprising a pyrano[2,3-f]chromene derivative compound of the following chemical formula 1, a pharmaceutically acceptable salt thereof, or a solvate thereof:

[0029] [Chemical Formula 1]

[0030]

[0031] In the above chemical formula 1, R1 is a hydrogen atom; a substituted or unsubstituted, straight-chain or branched C1-C6 alkyl group; a halogen atom; a substituted or unsubstituted, straight-chain or branched C1-C6 alkoxy group; or a substituted or unsubstituted, straight-chain or branched C1-C4 thioalkyl group; and R2 is a hydrogen atom, methyl, ethyl, methoxy, or ethoxy, and R3 and R4 are each independently a hydrogen atom or a C1-C2 alkyl group, and in the case of the substituted alkyl, substituted alkoxy, and substituted thioalkyl, the substituent is a straight-chain or branched C1-C5 alkyl group, a halogen atom, a straight-chain or branched C1-C5 alkoxy group, or a straight-chain or branched C1-C3 thioalkyl group.

[0032] A pharmaceutical composition comprising a pyrano[2,3-f]chromene derivative compound, a pharmaceutically acceptable salt thereof, or a solvate thereof according to one embodiment of the present invention has excellent preventive and therapeutic effects on degenerative osteoarthritis, particularly primary osteoarthritis.

[0033] In one embodiment of the present invention, the degenerative osteoarthritis may include osteoporosis, knee arthritis, and / or knee pain. In addition, the degenerative osteoarthritis may be primary osteoarthritis or secondary osteoarthritis. The primary osteoarthritis may be primary osteoarthritis or osteoarthritis caused by aging, obesity, injury, etc. The secondary osteoarthritis may be chondromalacia patellae or patellofemoral degenerative arthritis among the primary osteoarthritis.

[0034] A pharmaceutical composition comprising a pyrano[2,3-f]chromene derivative compound according to the present invention, a pharmaceutically acceptable salt thereof, or a solvate thereof has excellent preventive and therapeutic efficacy against osteoarthritis that can cause secondary osteoarthritis, such as primary osteoarthritis, chondromalacia patellae, and patellofemoral degenerative arthritis.

[0035] In the present invention, the degenerative osteoarthritis is a disease in which inflammation and pain occur due to damage to bones and ligaments forming the joint caused by gradual damage or degenerative changes in the cartilage protecting the joint.

[0036] According to one embodiment of the present invention, in the chemical formula 1, R1 may be a substituted or unsubstituted, straight-chain or branched C1-C6 alkoxy group. Specifically, R1 may be an unsubstituted straight-chain or branched C1-C6 alkoxy group. In addition, in the chemical formula 1, R2 may be a hydrogen atom, and R3 and R4 may be a C1-C2 alkyl group. When R1 to R5 in the chemical formula 1 are as described above, a pharmaceutical composition comprising the pyrano pyrano[2,3-f]chromene derivative compound of the chemical formula 1, a pharmaceutically acceptable salt thereof, or a solvate thereof is chemically stable and has excellent preventive and therapeutic efficacy for degenerative osteoarthritis, particularly primary osteoarthritis.

[0037] According to one embodiment of the present invention, the compound represented by the chemical formula 1 may include any one of the following compounds.

[0038]

[0039]

[0040] The pharmaceutical composition comprising any one of the compounds 1 to 16, a pharmaceutically acceptable salt thereof, or a solvate thereof is chemically stable and has excellent preventive and therapeutic efficacy for degenerative osteoarthritis, particularly primary osteoarthritis.

[0041] In addition, the pharmaceutical composition may have an excellent preventive or therapeutic effect on degenerative osteoarthritis even when administered once a day. Furthermore, the pharmaceutical composition may achieve an excellent anti-degenerative osteoarthritis effect even when the active ingredient is administered in a relatively small dosage. In particular, the pharmaceutical composition comprising any one of Compounds 1 to 16 may have a superior anti-degenerative osteoarthritis effect compared to a pharmaceutical composition comprising any one of Compounds 17 to 28 described below.

[0042] According to one embodiment of the present invention, the compound represented by the chemical formula 1 may include any one of the following compounds.

[0043]

[0044]

[0045] The pharmaceutical composition comprising any one of the compounds 17 to 28, a pharmaceutically acceptable salt thereof, or a solvate thereof can exhibit excellent anti-degenerative osteoarthritis effects.

[0046] A pharmaceutical composition comprising the pyrano[2,3-f]chromene derivative compound of the above chemical formula 1, a pharmaceutically acceptable salt thereof, or a solvate thereof has excellent preventive and therapeutic effects on degenerative osteoarthritis, particularly primary osteoarthritis.

[0047] According to one embodiment of the present invention, the pyranochromene derivative compound of Chemical Formula 1 suppresses chemokines and cytokines that induce an inflammatory response, such as IL-6 and MCP-1, in the cartilage tissue of the knee joint, and induces an anti-inflammatory response in the cartilage tissue of the joint by suppressing MMP3, a cartilage-degrading protein, and RIPK3, which is involved in cartilage cell apoptosis. Therefore, it can have an excellent effect in the prevention or treatment of degenerative osteoarthritis, especially primary osteoarthritis.

[0048] According to one embodiment of the present invention, a pharmaceutically acceptable salt of the compound of formula 1 may mean that the compound of formula 1 forms a salt with a free acid and exists as an acid addition salt. The compound of formula 1 can form a pharmaceutically acceptable acid addition salt according to a conventional method known in the art. An organic acid or an inorganic acid may be used as the free acid, and the inorganic acid may be hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, or the like, and the organic acid may be citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, galacturonic acid, embonic acid, glutamic acid, aspartic acid, or the like.

[0049] A pharmaceutical composition according to one embodiment of the present invention may include, in addition to the pyrano[2,3-f]chromene derivative compound of the above chemical formula 1, a pharmaceutically acceptable salt thereof, or a solvate thereof, a pharmaceutically acceptable carrier, excipient, or diluent. Examples of the carrier, excipient, and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0050] In one embodiment of the present invention, the pyrano[2,3-f]chromene derivative compound of Chemical Formula 1 can be prepared by the same method as that described in Korean Patent Publication Nos. 10-2015-0075030 and 10-2018-0002539. However, the present invention is not limited thereto.

[0051] According to one embodiment of the present invention, the pharmaceutical composition may be formulated into a conventional pharmaceutical formulation known in the art. The formulation may be formulated and administered in any dosage form, including, but not limited to, oral administration formulations, injections, suppositories, transdermal administration formulations, and nasal administration formulations. Preferably, the composition may be formulated into oral administration formulations and injections.

[0052] When formulating into each of the above dosage forms, a pharmaceutically acceptable carrier necessary for the manufacture of each dosage form may be added to manufacture the composition. The term "pharmaceutically acceptable carrier" is used herein to refer to any component other than a pharmaceutically active ingredient. "Pharmaceutically acceptable" refers to a property that does not cause pharmaceutically undesirable changes by interacting with other components present in the composition (e.g., interaction between carriers or between the pharmaceutically active ingredient and the carrier). The selection of the pharmaceutically acceptable carrier may vary depending on factors such as the characteristics of a specific dosage form, the administration method, and the effect of the carrier on solubility and stability.

[0053] According to one embodiment of the present invention, a pharmaceutically acceptable carrier included in a pharmaceutical composition for oral administration may be at least one selected from, but is not limited to, a diluent, a binder, a disintegrant, a glidant (or a lubricant), an adsorbent, a stabilizer, a solubilizer, a sweetener, a colorant, and a flavoring agent.

[0054] A diluent refers to any excipient added to increase the volume of a composition to make it an appropriate size according to the dosage form. The diluent may be at least one selected from, but is not limited to, starch (e.g., potato starch, corn starch, wheat starch, pregelatinized starch), microcrystalline cellulose (e.g., low-hydration microcrystalline cellulose), lactose (e.g., lactose monohydrate, anhydrous lactose, spray lactose), glucose, sorbitol, mannitol, sucrose, alginate, alkaline earth metal salt, clay, polyethylene glycol dicalcium phosphate, anhydrous calcium hydrogen phosphate, and silicon dioxide. In the present invention, the diluent may be used in an amount of 5 wt% to 50 wt% based on the total amount of the pharmaceutical composition. For example, it may be used in an amount of 10 wt% to 35 wt% based on the total amount of the composition for tabletting and quality maintenance.

[0055] A binder refers to a substance used to impart adhesiveness to a powdered substance to facilitate compression and improve fluidity. The binder may be at least one selected from, but is not limited to, starch, microcrystalline cellulose, highly dispersible silica, mannitol, lactose, polyethylene glycol, polyvinylpyrrolidone, cellulose derivatives (e.g., hydroxypropylmethylcellulose, hydroxypropylcellulose, low-substituted hydroxypropylcellulose), natural gum, synthetic gum, povidone, copovidone, and gelatin. In the present invention, the binder may be used in an amount of 2 wt% to 15 wt% based on the total amount of the pharmaceutical composition. For example, it may be used in an amount of 1 wt% to 3 wt% for tableting and quality maintenance.

[0056] A disintegrant refers to a substance added to facilitate the disintegration or disintegration of a solid dosage form after administration to a body. The disintegrant may be used alone or in combination with, but is not limited to, starch or modified starch such as sodium starch glycolate, corn starch, potato starch or pregelatinized starch; clay such as bentonite, montmorillonite or veegum; cellulose such as microcrystalline cellulose, hydroxypropyl cellulose or carboxymethyl cellulose; alginates such as sodium alginate or alginic acid; cross-linked cellulose such as croscarmellose sodium; gums such as guar gum or xanthan gum; cross-linked polymers such as cross-linked polyvinylpyrrolidone (crospovidone); and effervescent agents such as sodium bicarbonate or citric acid. In the present invention, the disintegrant may be used in an amount ranging from 2% to 15% by weight based on the total amount of the pharmaceutical composition. For example, it may be used in an amount ranging from 4% to 10% by weight for tableting and quality maintenance.

[0057] A glidant or lubricant refers to a substance that functions to prevent powder from sticking to the pressing equipment and improve the flow of granules. The glidant may be, but is not limited to, light anhydrous silicic acid, talc, stearic acid, metal salts of stearic acid (magnesium salt or calcium salt, etc.), sodium lauryl sulfate, hydrogenated vegetable oil, sodium benzoate, sodium stearyl fumarate, glyceryl behenate, glyceryl monostearate, or polyethylene glycol, used alone or in combination.

[0058] In the present invention, the lubricant may be used in an amount of 0.1 wt% to 5 wt% based on the total amount of the pharmaceutical composition. For example, it may be used in an amount of 1 wt% to 3 wt% for tableting and quality maintenance.

[0059] The adsorbent may be, but is not limited to, hydrous silicon dioxide, light anhydrous silicic acid, colloidal silicon dioxide, magnesium aluminate metasilicate, microcrystalline cellulose, lactose, or cross-linked polyvinylpyrrolidone, used alone or in combination.

[0060] The stabilizer may be at least one selected from, but is not limited to, an antioxidant such as butylhydroxyanisole, butylhydroxytoluene, carotene, retinol, ascorbic acid, tocopherol, tocopherol polyethylene glycol succinic acid or propyl gallate, a cyclic compound of a sugar such as cyclodextrin, carboxyethyl cyclodextrin, hydroxypropyl cyclodextrin, sulfobutyl ether or cyclodextrin, and an organic acid such as phosphoric acid, lactic acid, acetic acid, citric acid, tartaric acid, succinic acid, maleic acid, fumaric acid, glycolic acid, propionic acid, gluconic acid or glucuronic acid.

[0061] Optionally, the pharmaceutical composition may include known additives to enhance palatability by enhancing taste. For example, sweeteners such as sucralose, sucrose, fructose, erythritol, acesulfame potassium, sugar alcohols, honey, sorbitol, or aspartame can be added to more effectively mask the bitter taste and maintain the stability and quality of the formulation. Additionally, acidulants such as citric acid and sodium citrate, natural flavors such as plum, lemon, pineapple, and herbal flavors, and natural fruit juices, chlorophyllin, and flavonoids can be used.

[0062] The pharmaceutical composition for oral administration may be a solid preparation, a semi-solid preparation, or a liquid preparation for oral administration. Examples of solid preparations for oral administration include, but are not limited to, tablets, pills, hard or soft capsules, powders, granules, granules, powders for reconstitution of solutions or suspensions, lozenges, wafers, oral strips, dragees, chewable gums, and the like. Liquid preparations for oral administration include solutions, suspensions, emulsions, syrups, elixirs, spirits, flavorings, lemonades, extracts, precipitants, tinctures, and emulsions. Semi-solid preparations include, but are not limited to, aerosols, creams, gels, and the like. The pharmaceutical composition according to the present invention may be formulated as an injection, and when formulated as an injection, it may include blood and a non-toxic buffer solution as a diluent, for example, a phosphate buffer solution having a pH of 7.4. The pharmaceutical composition may include other diluents or additives in addition to the buffer solution.

[0063] The carrier used in the above-mentioned preparation and the method for preparing the preparation can be selected and prepared according to those widely known in the art, and can be prepared, for example, according to the method described in the latest edition of Remington's Pharmaceutical Science.

[0064] The dosage and timing of administration of the pharmaceutical composition according to the present invention may vary depending on the age, sex, condition, weight, administration route, number of administrations, and form of the drug of the subject. The daily dosage is about 0.1 to 300 mg / kg, preferably 1 to 200 mg / kg, based on the active ingredient, i.e., the compound of Chemical Formula 1. The dosage may be appropriately increased or decreased depending on the type of disease, the degree of disease progression, the administration route, sex, age, weight, etc.

[0065] The pharmaceutical composition according to the present invention may be administered in several divided doses so that the total daily dose for adults is 1 to 100 mg / kg based on the compound of chemical formula 1 as the active ingredient in order to obtain the desired effect, and specifically, it may be administered in several divided doses so that it is 25 to 75 mg / kg, and more specifically, it may be administered in several divided doses so that it is 50 mg / kg.

[0066] As one specific example of the invention, the pharmaceutical composition according to the present invention can reduce an inflammatory factor, specifically, the inflammatory factor of the pharmaceutical composition can be reduced by 75 to 25%, and more specifically, can be reduced by 50%.

[0067] One embodiment of the present invention provides a method for preventing or treating degenerative osteoarthritis, comprising administering to a subject suspected of having degenerative osteoarthritis a composition comprising a pyrano[2,3-f]chromene derivative compound of the following chemical formula 1, a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0068] [Chemical Formula 1]

[0069]

[0070] In the above chemical formula 1, R1 is a hydrogen atom; a substituted or unsubstituted, straight-chain or branched C1-C6 alkyl group; a halogen atom; a substituted or unsubstituted, straight-chain or branched C1-C6 alkoxy group; or a substituted or unsubstituted, straight-chain or branched C1-C4 thioalkyl group; and R2 is a hydrogen atom, methyl, ethyl, methoxy, or ethoxy, and R3 and R4 are each independently a hydrogen atom or a C1-C2 alkyl group, and in the case of the substituted alkyl, substituted alkoxy, and substituted thioalkyl, the substituent is a straight-chain or branched C1-C5 alkyl group, a halogen atom, a straight-chain or branched C1-C5 alkoxy group, or a straight-chain or branched C1-C3 thioalkyl group.

[0071] One embodiment of the present invention provides a composition comprising a pyrano[2,3-f]chromene derivative compound of the following chemical formula 1, a pharmaceutically acceptable salt thereof, or a solvate thereof for the prevention or treatment of degenerative osteoarthritis.

[0072] [Chemical Formula 1]

[0073]

[0074] In the above chemical formula 1, R1 is a hydrogen atom; a substituted or unsubstituted, straight-chain or branched C1-C6 alkyl group; a halogen atom; a substituted or unsubstituted, straight-chain or branched C1-C6 alkoxy group; or a substituted or unsubstituted, straight-chain or branched C1-C4 thioalkyl group; and R2 is a hydrogen atom, methyl, ethyl, methoxy, or ethoxy, and R3 and R4 are each independently a hydrogen atom or a C1-C2 alkyl group, and in the case of the substituted alkyl, substituted alkoxy, and substituted thioalkyl, the substituent is a straight-chain or branched C1-C5 alkyl group, a halogen atom, a straight-chain or branched C1-C5 alkoxy group, or a straight-chain or branched C1-C3 thioalkyl group.

[0075] The pharmaceutical composition according to the present invention may contain the compound of the chemical formula I according to the present invention in an amount of about 1 to 50 wt%, preferably 10 to 40 wt%, based on the total weight of the entire composition.

[0076]

[0077] Hereinafter, the present invention will be described in detail using examples. However, the examples according to the present invention may be modified in various ways, and the scope of the present invention is not limited to the examples described below. The examples in this specification are provided to more fully explain the present invention to those of ordinary skill in the art.

[0078]

[0079]

[0080] Experimental Example 1: Confirmation of pain suppression effect in an OA rat model using MIA.

[0081]

[0082] Experimental Example 1-1. Experimental Preparation

[0083] All animals were allowed to acclimatize to the laboratory environment for one week after introduction. On the day of the experiment, monosodium iodoacetate (MIA, Sigma, Cat. No. I2512) was dissolved in saline for injection at a concentration of 30 mg / mL, and 50 μL was administered once on the first day of the experiment. Animal groups were divided as follows. Animals were randomly distributed according to the pain threshold evaluation scores ranked by rank to ensure that the average of each group was distributed as evenly as possible, resulting in the numbers specified in the "Composition of Test Groups" table. Individual identification was performed by directly writing numbers on the animals' tails. Colored identification cards for each dose were attached to the cages. An animal room usage record sheet was attached to the entrance of the room, containing the study number, period of animal room use, name of the study director, name of the study staff, and emergency contact information. The test group consisted of WT, Vehicle, Compound 2 25 mg / kg, and Compound 2 50 mg / kg, and was administered orally once a day for a total of 21 days starting from 3 days after MIA administration.

[0084] The nociceptive threshold (g) was measured using a dynamic plantar aesthesiometer (Ugo Basile, 37400, Comerio, Italy), a device that gradually increases the force applied to the animal's paw over a set period of time. Before measurement, the animals were placed in an acrylic box with a wire mesh bench below and allowed to acclimate for 5 minutes. The pain threshold was measured by gradually applying a force from 0 to 50 g over 10 seconds to the center of the right hind paw using a metal filament. The weight at which the animal elicited a paw withdrawal behavior was measured. The cut-off threshold was set at 50 g to avoid tissue damage. Measurements were taken before arthritis induction (Day 0) to obtain a baseline value, and then before test substance administration (Day 3) and at fixed times twice a week after intra-articular test substance administration. The pain threshold results were calculated as paw withdrawal latency (sec) and paw withdrawal threshold (g).

[0085]

[0086] Experimental Example 1-2. Results from the experiment

[0087] As a result of measuring pain due to cartilage damage in an OA animal model according to Experimental Example 1-1, it was confirmed that pain in the OA model was gradually reduced as compound 2 was administered. It was confirmed that it exhibited efficacy equivalent to that of the positive control, celecoxib (Fig. 1).

[0088]

[0089]

[0090] Experimental Example 2: Confirmation of improved weight bearing in an OA rat model using MIA.

[0091]

[0092] Experimental Example 2-1. Preparation for an experiment to improve weight bearing.

[0093] Weight bearing was measured from the same model as Experimental Example 1. The weight bearing measurement test was an experiment to measure the change in weight bearing (or weight distribution) between the normal hind limb (left) and the arthritic hind limb (right) caused by pain in the hind limb knee induced by unilateral arthritis. The load of each hind paw was measured using an incapacitance meter (Model 600, IITC, USA), a device for measuring weight bearing. Since the load may change depending on the animal's foot-stepping posture, the animal was positioned accurately in the holder so that both feet could step symmetrically, and only one person was responsible for fixing it to minimize errors. Before the measurement, the animal was placed in an acrylic box and allowed to acclimate for 5 minutes.

[0094] When each animal was correctly positioned in the holder, the machine was operated, and the measurement time was set to 5 seconds, and a total of two measurements were taken, and the average value was taken as the weight bearing (g) value of each paw. Measurements were taken before arthritis induction (Day 0) to obtain a baseline value, then before test substance administration (Day 3), and twice a week at a set time after intra-articular test substance administration. The results of the weight bearing measurement test were converted to weight bearing ratio using the following calculation formula and analyzed.

[0095]

[0096] Experimental Example 2-2. Results from Experiment 2-1

[0097] As a result of measuring the weight load according to cartilage damage in the OA animal model according to Experimental Example 2-1, it was confirmed that the weight load in the OA model was reduced by administering compound 2.

[0098] Additionally, it can be confirmed that it shows efficacy equivalent to that of the positive control group, celecoxib (Figure 2).

[0099]

[0100]

[0101] Experimental Example 3: Histopathological analysis according to therapeutic effects in an MIA-OA animal model.

[0102]

[0103] Experimental Example 3-1. Histopathological analysis according to therapeutic effect in MIA-OA animal model

[0104] The knee joint was analyzed histopathologically using the same model as Experimental Example 1. After histopathological analysis, two evaluators scored the severity of osteoarthritis according to the following evaluation criteria after Safranin O staining. The analysis criteria were analyzed using the OARSI and Total Mankin score. The OARSI analysis criteria were scored as follows: 0: Normal cartilage surface; 1: Normal cartilage surface with cell death; 2: Disconnection of the cartilage surface and fibrosis of the superficial layer; 3: Cracking along the longitudinal axis of the cartilage; 4: Damage and infiltration up to the middle of the cartilage; 5: Exposure of the stromal bone area; 6: Complete cartilage damage and osteophyte formation. The Total Mankin score analysis criteria were analyzed according to four items: Structure (0-6), Cell (0-3), Safranin O (0-4), and Tidemark (0-1). A higher score for each item indicates more damage to cartilage cells and structure.

[0105]

[0106] Experimental Example 3-2. Experimental Results According to Histopathological Analysis Method

[0107] As a result of confirming the tissue pathology of the knee joint in the OA animal model according to Experimental Example 3-1, Safranin 0 stained in the cartilage tissue of the OA model was improved by administering compound 2 (Fig. 3).

[0108] Additionally, ORASI and Mankin scores, which analyze the severity of osteoarthritis and assign scores, were reduced. This confirms that the drug exhibits efficacy equivalent to that of the positive control, celecoxib (Figure 4).

[0109]

[0110]

[0111] Experimental Example 4: Tissue immunochemical analysis according to therapeutic effect in MIA-OA animal model

[0112]

[0113] Experimental Example 4-1. Tissue immunochemical analysis in an MIA-OA animal model

[0114] Knee joint tissue pathology analysis was performed using the same model as Experimental Example 1. For immunochemical analysis of joint tissue, specific IL-6, MCP-1, MMP3, and RIPK3 antibodies were used to stain the joint tissue, and photographs were taken under a microscope at 400x magnification. Then, using ImageJ software (Wayne Rasband, NIH, USA), the Dab positive area expressing each cytokine in the synovium was extracted and analyzed as a % compared to the total nuclear area.

[0115]

[0116] Experimental Example 4-2. Results of tissue immunochemical analysis according to the therapeutic effect in the MIA-OA animal model.

[0117] As a result of tissue immunochemical analysis of the knee joint in the OA animal model according to Experimental Example 4-1, it was confirmed that IL-6 and MCP-1, which are proteins that cause inflammation in the OA model, were suppressed by administering compound 2 (Figs. 5 and 6).

[0118] Additionally, it was confirmed that MMP3, a protein that decomposes cartilage, was reduced, and RIPK3, which is involved in cartilage cell death, was inhibited. At this time, it was confirmed that it showed superior efficacy compared to the positive control group, celecoxib (Figs. 7, 8).

[0119] The present invention has been described above, focusing on preferred embodiments thereof. Those skilled in the art will appreciate that the present invention can be implemented in modified forms without departing from its essential characteristics. Therefore, the disclosed embodiments should be considered illustrative rather than limiting. The scope of the present invention is set forth in the claims, not the foregoing description, and all differences within the scope equivalent thereto should be construed as being encompassed by the present invention.

[0120]

[0121] The present invention aims to provide a pharmaceutical composition for preventing or treating degenerative arthritis. Specifically, the present invention relates to a pharmaceutical composition for preventing or treating degenerative arthritis comprising a pyrano[2,3-f]chromene derivative compound, a pharmaceutically acceptable salt thereof, or a solvate thereof, and therefore has industrial applicability in terms of treating, improving, alleviating, and preventing degenerative arthritis.

Claims

1. A pharmaceutical composition for preventing or treating degenerative osteoarthritis, comprising a pyrano[2,3-f]chromene derivative compound of the following chemical formula 1, a pharmaceutically acceptable salt thereof, or a solvate thereof: [Chemical Formula 1] In the above chemical formula 1, R1 is a hydrogen atom; a substituted or unsubstituted, straight-chain or branched C1-C6 alkyl group; a halogen atom; a substituted or unsubstituted, straight-chain or branched C1-C6 alkoxy group; or a substituted or unsubstituted, straight-chain or branched C1-C4 thioalkyl group; R2 is a hydrogen atom, methyl, ethyl, methoxy or ethoxy, R3 and R4 are each independently a hydrogen atom or a C1-C2 alkyl group, In the case of the above substituted alkyl, substituted alkoxy and substituted thioalkyl, the substituent is a straight or branched C1-C5 alkyl group, a halogen atom, a straight or branched C1-C5 alkoxy group or a straight or branched C1-C3 thioalkyl group.

2. In paragraph 1, The above R1 is ethoxy, n-propoxy, isopropoxy, n-butoxy, 2-methylpropoxy, n-pentoxy, 2-methylbutoxy, 3-methylbutoxy, 2-ethylpropoxy, n-hexyloxy, 2-methylpentoxy, 3-methylpentoxy, 4-methylpentoxy or 2-ethylbutoxy; R2 is a hydrogen atom, R3 and R4 are methyl, A pharmaceutical composition for the prevention or treatment of degenerative osteoarthritis.

3. In paragraph 1, The compound represented by the above chemical formula 1 comprises any one of the following compounds: Pharmaceutical composition for the prevention or treatment of degenerative osteoarthritis:

4. In paragraph 1, A pharmaceutical composition for the prevention or treatment of degenerative osteoarthritis, wherein the above degenerative osteoarthritis is primary osteoarthritis or secondary osteoarthritis.

5. In paragraph 4, A pharmaceutical composition for the prevention or treatment of degenerative osteoarthritis, wherein the above primary osteoarthritis is primary osteoarthritis.

6. A pharmaceutical composition for preventing or treating degenerative osteoarthritis, wherein the secondary osteoarthritis is primary osteoarthritis such as chondromalacia or patellofemoral degenerative arthritis.

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