WO2025118014A1

WO2025118014A1 - A composition containing a carboxamide redox derivative compound as an active ingredient for the prevention or treatment of arthritis

Info

Publication number: WO2025118014A1
Application number: PCT/AU2024/051276
Authority: WO
Inventors: In Hyun Lee
Original assignee: Benobio Australia Pty Ltd; Benobio Co Ltd
Current assignee: Benobio Australia Pty Ltd; Benobio Co Ltd
Priority date: 2021-11-30
Filing date: 2024-11-29
Publication date: 2025-06-12
Anticipated expiration: 2026-06-04
Also published as: KR102619332B1; KR102619331B1; KR20230081959A; KR20230081602A; KR20230169049A

Abstract

Provided is a composition for the prevention or treatment of arthritis including a compound represented by the following Chemical Formula I or a pharmaceutically acceptable salt thereof as an active ingredient.

Description

DESCRIPTION

Invention Title

A COMPOSITION CONTAINING A CARBOXAMIDE REDOX DERIVATIVE COMPOUND AS AN ACTIVE INGREDIENT FOR THE PREVENTION OR TREATMENT OF ARTHRITIS

Technical Field

The present invention relates to a pharmaceutical composition or food composition containing a carboxamide redox derivative compound having cartilage tissue destruction inhibition ability and cartilage regeneration ability as an active ingredient for the prevention or treatment of arthritis.

Background Art

Bone tissue generally consists of compact bone osteoid with a strong bone surface, and sponge bone osteoid in which the center or both ends of the long bone has osteoid joined like a mesh. Most bones are initially developed as cartilage in a connective tissue, which later changes into bone tissue, and some bones are directly formed in a connective tissue.

Cartilage is an elastic tissue that connects two bones, provides mobility to the articular area, and plays a role in shock absorption. In particular, articular cartilage is a connective tissue and includes cartilage cells embedded in the cartilage matrix, which is an intercellular substance containing proteoglycan, collagen fibrils such as type II collagen, other proteins, and water. Damage to the cartilage tissue may cause arthritis and the like, which causes direct friction between bones, stiffness of joints, a gradual decrease in articular cartilage movement, and frequent pain in the cartilage area. In the bone surface, there is a tough connective tissue periosteum, and nerves and blood vessels are distributed to protect and nourish the bone. If the periosteum is damaged, bone survival, new growth, and regeneration become difficult. The bone osteoid component is composed of water, an organic material including cells, and an inorganic material, and the constant elasticity of the bone is due to the organic material. As the age increases, the inorganic material (mainly calcium phosphate) increases, thereby increasing bone hardness.

Arthritis is a type of inflammatory disease, and may be broadly divided into chronic inflammation, rheumatoid arthritis, and degenerative arthritis. The exact cause and mechanism of the chronic inflammation, rheumatoid arthritis, are not yet well known, but one of the strongest causes that have been discovered so far is known to occur when bone homeostasis is disrupted due to a numerical imbalance between osteoblasts that generate bone and osteoclasts that destroy bone. Normal osteoclasts are differentiated from monocytes and macrophages and play a role in destroying old bone or removing damaged bone, but the appearance of osteoclasts found in patients with rheumatoid arthritis shows a pathological form in which bone is damaged due to a numerical increase, continuous survival, and excessive activity caused by osteoclatogeneration. The degenerative arthritis is a disease in which inflammation and pain occur due to damage to the bones and ligaments that form the joint, caused by gradual damage or degenerative changes in the cartilage that protects the joint. The degenerative arthritis has the highest frequency among inflammatory diseases of the joints, and the cartilage tissue in joints such as the knee, ankle, fingers, elbow, and hip joints is gradually degenerated and destroyed due to old age, obesity, joint trauma, joint dysplasia, a history of arthritis, some special occupations, genetic predisposition, and the like. Meanwhile, despite the diversity of causes of arthritis, there is no therapeutic agent that may directly resolve the cause of the disease, and there are only methods for relieving pain or delaying the severity of an already occurring disease, and thus there is a need for the development of a therapeutic agent that is effective in fundamentally treating arthritis.

[Prior Arts]

[Patent Documents]

Korean Patent Registration No. 10-2127887

Disclosure

Technical Problem

The present invention has been made in an effort to provide a composition for the prevention or treatment of arthritis, including a carboxamide redox derivative compound having cartilage tissue regeneration ability and cartilage tissue destruction inhibition ability.

Technical Solution

In order to solve the above problem, one aspect of the present invention provides a pharmaceutical composition or food composition for the prevention or treatment of arthritis, including a compound represented by the following Chemical Formula I or a pharmaceutically acceptable salt thereof as an effective ingredient:

[Chemical Formula I]

Advantageous Effects

According to the present invention, the pharmaceutical composition or food composition for the prevention or treatment of arthritis including the carboxamide redox derivative compound or the pharmaceutically acceptable salt thereof as the active ingredient has an excellent effect on cartilage tissue regeneration and cartilage tissue destruction inhibition. Further, according to the composition including the carboxamide redox derivative compound or the pharmaceutically acceptable salt thereof as the active ingredient, there are effects of reducing pain in arthritis patients, having increased walking ability and a cartilage regeneration effect, and improving arthritis.

Description of Drawings

FIG. 1 illustrates results of 'H-NMR spectrum analysis of a compound of Chemical Formula I according to an example of the present invention. FIG. 2 illustrates results of LCMS analysis of a compound of Chemical

Formula I according to an example of the present invention.

FIG. 3 illustrates results of confirming cartilage destruction inhibition ability of a composition according to an example of the present invention using Safranin O- staining. FIG. 4 illustrates results of measuring OASRSI grade and SBP thickness of a composition according to an example of the present invention.

FIG. 5 illustrates results of confirming cartilage destruction inhibition ability of a composition according to an example of the present invention using H&E and Safranin O-staining.

FIG. 6 illustrates results of confirming cartilage regeneration ability of a composition according to an example of the present invention using immunohistochemical staining (Collagen type 2).

Modes of the Invention

Hereinafter, various examples of the present invention will be described in detail with reference to the accompanying drawings. The present invention is not limited to specific examples, and it should be understood to include various modifications, equivalents, and/or alternatives to the examples of the present invention. In connection with the description of the drawings, similar reference numerals may be used for similar components.

In this specification, expressions such as “have”, “may have”, “include”, or “may include” refer to the presence of the corresponding feature (e.g., numerical value, function, operation, or component such as part), and does not exclude the presence of additional features.

In the present invention, the expression such as “A or B”, “at least one of A and/or B”, or “one or more of A and/or B” may include all possible combinations of items listed together. For example, “A or B”, “at least one of A and B”, or “at least one of A or B” may refer to all cases of (1) including at least one A, (2) including at least one B, or (3) including both at least one A and at least one B. The expression “configured to” used in this specification may be changed and used to, for example, “suitable for”, “having the capacity to”, “designed to”, “adapted to”, “made to” or “capable of’, depending on the situation. The term of “configured to” may not necessarily mean “specially designed to.”

The terms used in this specification are used to illustrate only specific examples, and may not be intended to limit the scope of other examples. A singular expression may include a plural expression unless the context clearly indicates otherwise. The terms used herein, including technical or scientific terms, may have the same meaning as generally understood by those of ordinary skill in the art described in the present invention. The terms defined in a general dictionary among the terms used in this specification may be interpreted in the same or similar meaning as or to the meaning on the context of the related art, and will not be interpreted as an ideal or excessively formal meaning unless otherwise defined in the present invention. In some cases, even the terms defined in the present invention may not be interpreted to exclude the examples of the present invention.

The examples disclosed in the present invention are presented for explanation and understanding of the disclosed technical contents, and do not limit the scope of the present invention. Therefore, the scope of the present invention should be interpreted as including all changes or various other examples based on the technical idea of the present invention.

Hereinafter, a preferred example of the present invention will be described in detail. Terms and words used in the present specification and claims should not be interpreted as being limited to typical or dictionary meanings, but should be interpreted as having meanings and concepts which comply with the technical spirit of the present invention, based on the principle that an inventor may appropriately define the concept of the term to describe his/her own invention in the best manner.

Therefore, the configurations of the examples described in the present specification are merely the most preferred example of the present invention and are not intended to represent all of the technical ideas of the present invention, and thus, it should be understood that there are various equivalents and modifications capable of replacing the configurations at the time of this application.

Throughout the specification, when a part “includes” a component, unless otherwise specifically stated for the contrary, it is meant to further include other components rather than excluding other components.

Hereinafter, the present invention will be described in detail.

According to an example of the present invention, there is provided a pharmaceutical composition for the prevention or treatment of arthritis, including a compound represented by the following Chemical Formula I, an isomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient:

[Chemical Formula I]

The compound represented by Chemical Formula I is N-methyl-2-(l-((5-(2-(2- methyl-l-oxoisoindolin-4-yl)phenyl)furan-2-yl)methyl)-3-oxopiperazin-2- yl)acetamide. In the present disclosure, the term ‘isomer’ means a compound of the present invention or a salt thereof having the same Chemical Formula or Molecular Formula, but being structurally or sterically different. These isomers include structural isomers such as tautomers, and stereoisomers, and the stereoisomers include R or S isomers (optical isomers, enantiomers) having an asymmetric carbon center, geometric isomers (trans, cis), etc. In the present invention, all stereoisomers of the compound represented by Chemical Formula I and mixtures thereof are also included in the scope of the present invention.

In the present disclosure, the term ‘solvate’ means that the compound represented by Chemical Formula I and a solvent other than water are bound by non- covalent intermolecular force, and includes a stoichiometric or non-stoichiometric amount of solvent. Preferred solvents are volatile and non-toxic, and may be administered to humans in very small amounts. Specifically, the solvate may include water in a ratio of about 0.25 mole to about 10 moles based on 1 mole of the active ingredient, and more specifically, about 0.5 mole, about 1 mole, about 1.5 moles, about 2 moles, about 3 moles, about 5 moles, etc.

In the present disclosure, the term ‘pharmaceutically acceptable salt’ refers to a salt commonly used in the pharmaceutical industry. For example, the pharmaceutically acceptable salt includes inorganic ion salts prepared from calcium, potassium, sodium, magnesium, etc.; inorganic acid salts prepared from hydrochloric acid, nitric acid, phosphoric acid, hydrobromic acid, iodic acid, perchloric acid, sulfuric acid, etc.; organic acid salts prepared from acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, etc.; sulfonic acid salts prepared from methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p- toluenesulfonic acid, naphthalenesulfonic acid, etc.; and amine salts prepared from trimethylamine, triethylamine, ammonia, pyridine, picoline, etc. However, the types of salt in the present invention are not limited to these listed salts.

In the present disclosure, the term ‘including as the active ingredient’ means containing in a dose range that brings about an effect of preventing, improving, or treating arthritis, and the dose range may vary depending on the severity and formulation, and the number of applications may also vary depending on the age, weight, and constitution of a subject of application. In an example of the present invention, the compound represented by Chemical Formula I in the pharmaceutical composition of the present invention may be, for example, 0.001 mg/kg or more, preferably 0.002 mg/kg or more. The quantitative upper limit of the compound represented by Chemical Formula I included in the pharmaceutical composition of the present invention may be selected and implemented within an appropriate range by those skilled in the art.

The pharmaceutical composition according to the present invention may include an effective amount of the compound represented by Chemical Formula I alone, or may include one or more pharmaceutically acceptable carriers, excipients, or diluents.

The pharmaceutically acceptable carrier, excipient, or diluent refers to a material that is physiologically acceptable and does not cause an allergic reaction, such as gastrointestinal disorder, dizziness, etc., or a similar reaction thereto when administered to humans. Examples of the carrier, the excipient, and the diluent may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil, but are not limited thereto. In addition, the pharmaceutical composition may further include fillers, anticoagulating agents, lubricants, wetting agents, flavorings, emulsifiers, preservatives, and the like.

The pharmaceutical composition of the present invention may be formulated by using a method known in the art so as to provide rapid, sustained, or delayed release of the active ingredient after administrated to the subject. The pharmaceutical composition may be formulated in the form of an oral preparation, an injectable preparation, or an external preparation. The oral preparation may be selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, oral solutions, emulsions, syrups, and lyophilized preparations, but is not limited thereto. In addition, the external preparation may be selected from the group consisting of creams, gels, ointments, emulsions, suspensions, sprays, and transdermal delivery patches, but is not limited thereto.

The pharmaceutical composition of the present invention may be administered orally or through other routes including transdermal, subcutaneous, intravenous or intramuscular route.

In the present disclosure, the term ‘prevention’ means preventing the occurrence of arthritis symptoms in advance by administering, ingesting, or applying the composition (e.g., pharmaceutical composition or food composition) of the present invention to a subject not suffering from arthritis to suppress or block the arthritis symptoms. In the present disclosure, the term ‘treatment’ includes not only complete cure of arthritis symptoms but also partial cure, improvement, and alleviation of arthritis symptoms, as a result of administering the composition (e.g., pharmaceutical composition or food composition) of the present invention to a subject suffering from arthritis.

In the present disclosure, the term ‘subject’ refers to all animals, including humans, that have already developed or may develop arthritis.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount.

In the present disclosure, the term ‘pharmaceutically effective amount’ refers to an amount enough to treat the disease at a reasonable benefit/risk ratio applicable to medical treatment or improvement, and the effective dose level may be determined according to factors including the type, severity, age, and sex of a subject, the activity of a drug, the sensitivity to a drug, a time of administration, a route of administration, an excretion rate, duration of treatment, and drugs to be simultaneously used, and other factors well-known in the medical field.

In the present disclosure, the term ‘administration’ means providing a predetermined material to a subject or patient by any suitable method, and the pharmaceutical composition may be administered parenterally (e.g., applied intravenously, subcutaneously, intraperitoneally or topically in injectable formulations) or orally according to a desired method, and the dose range may vary depending on the body weight, age, sex, and health condition of a patient, a diet, an administration time, an administration method, an excretion rate, and the severity of a disease. Specifically, in the present disclosure, the term ‘parenteral administration’ means a method of administering subcutaneously, intramuscularly, intravenously, or intraperitoneally using a tube, excluding oral administration. In addition, in the present disclosure, the term ‘oral administration’ means a method of injecting a drug into the mouth to improve pathological symptoms.

The composition may have cartilage tissue regeneration ability or cartilage tissue destruction inhibition ability.

In the cartilage tissue regeneration ability and the cartilage tissue destruction inhibition ability, ‘cartilage tissue’ is a flexible connective tissue found in many body parts of humans and animals, including bones, rib cage, ears, nose, bronchial tubes, and joints between intervertebral discs. The cartilage tissue is not as hard and rigid as bone, but us much stiffer and less flexible than muscle. Cartilage is composed of specialized cells called chondroblasts that produce a large amount of extracellular matrix consisting of collagen fibers, a proteoglycan-rich ground substance, and elastin fibers.

In the cartilage tissue regeneration ability, ‘regeneration’ generally refers to an action of restoring a part of the body or its function to its original state by regenerating the tissue or organ of the part when the part of the body or its function is lost in a living organism. In the cartilage tissue regeneration ability, the regeneration may be promoting the expression of type II collagen (Col II), which is a major component of the extracellular matrix (ECM) of cartilage cells, which is important for the production (synthesis) of cartilage cells. The type II collagen (Col II) maintains the elasticity of cartilage and acts as a support, and if the collagen is insufficient, the cartilage tissue may not be formed normally and becomes thin, which leads to erosion around the cartilage or arthritis. The cartilage tissue destruction inhibition ability may be evaluated by promoting the secretion of type II collagen (Col II) to prevent the destruction of cartilage and cartilage tissue. The arthritis may be at least one selected from the group consisting of osteoarthritis, rheumatoid arthritis, degenerative arthritis, psoriatic arthritis, and reactive arthritis, but is not limited thereto. Preferably, the arthritis may be at least one selected from the group consisting of osteoarthritis, rheumatoid arthritis and degenerative arthritis. More preferably, the arthritis may be osteoarthritis.

The compound represented by Chemical Formula I, the isomer thereof, the solvate thereof, or the pharmaceutically acceptable salt thereof may be contained in an amount of 0.00001 to 50 wt% based on the total weight of the composition. Preferably, the compound represented by Chemical Formula I, the isomer thereof, the solvate thereof, or the pharmaceutically acceptable salt thereof may be contained in an amount of 0.0001 to 30 wt%. However, the amount thereof may be increased or decreased according to the needs of a drug user, and may be increased or decreased appropriately depending on various factors such as diet, nutritional status, and the degree of damage to bone tissue or cartilage tissue, and thus is not limited to the range.

The dose of the pharmaceutical composition of the present invention may be determined by a specialist according to various factors, such as the condition, age, and body weight of a patient, the degree of cartilage damage, the degree of disease progression, and the like. In addition, the daily dose of the pharmaceutical composition per unit formulation or 1/2, 1/3 or 1/4 of the dose thereof is contained and may be administered 1 to 6 times a day.

According to another example of the present invention, there is provided a food composition for the prevention or treatment of arthritis including the compound represented by Chemical Formula I, an isomer thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof, as an active ingredient. The food includes health supplements, health functional foods, functional foods, etc., but is not limited thereto, and may include natural foods, processed foods, general food materials, etc. added with the compound represented by Chemical Formula I, the isomer thereof, the solvate thereof, or the pharmaceutically acceptable salt thereof. The food composition of the present invention may be added with the composition as it is or used with other foods or food compositions, and may be appropriately used according to a general method. The mixing amount of the active ingredients may be appropriately determined depending on the purpose of use.

The arthritis may be at least one selected from the group consisting of osteoarthritis, rheumatoid arthritis, degenerative arthritis, psoriatic arthritis, and reactive arthritis, but is not limited thereto. Preferably, the arthritis may be at least one selected from the group consisting of osteoarthritis, rheumatoid arthritis and degenerative arthritis. More preferably, the arthritis may be osteoarthritis.

The compound represented by Chemical Formula I, the isomer thereof, the solvate thereof, or the pharmaceutically acceptable salt thereof may be contained in an amount of 0.00001 to 50 wt% based on the total weight of the composition. Preferably, the compound represented by Chemical Formula I, the isomer thereof, the solvate thereof, or the pharmaceutically acceptable salt thereof may be contained in an amount of 0.0001 to 30 wt%. The effective dose of the compound represented by Chemical Formula I, the isomer thereof, the solvate thereof, or the pharmaceutically acceptable salt thereof may be used in accordance with the effective dose of the pharmaceutical composition. However, in the case of long-term intake for anti-inflammation, and improvement or maintenance of bone tissue or cartilage tissue damage, the effective dose may be below the range, and since the active ingredient has no problem in safety, the active ingredient may be used in an amount above the range. In addition, the food composition may further include one or more additives selected from the group consisting of an organic acid, a phosphate, an antioxidant, lactose casein, dextrin, glucose, sugar, and sorbitol, in addition to the compound represented by Chemical Formula I, the isomer thereof, the solvate thereof, or the pharmaceutically acceptable salt thereof.

Examples of the food may include meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, alcohol drinks, vitamin complex, other nutritional supplements, and the like, but are not limited to these types of foods.

Hereinafter, the present invention will be described in more detail through Examples. These Examples are only to describe the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these Examples in accordance with the gist of the present invention.

Preparation Examples and Examples

Compound I of the present invention was prepared according to the following Preparation Examples and Examples.

Preparation Example 1>

Preparation of 5-(2-(2-methyl-l-oxoisoindolin-4-yl)phenyl)furan-2- carboaldehyde

Preparation Example 1-1)

Preparation of 5-(2-bromophenyl)furan-2-carboaldehyde 2-bromoaniline (3.36 g, 35.0 mmol) was dissolved in a 2N-HC1 aqueous solution (50 ml), added dropwise with cuprous iodide (666 mg, 3.50 mmol) and then stirred at 0°C. At the same temperature, sodium nitrite (4.83 g, 70.0 mmol) was slowly added dropwise and stirred for 1 hour, and then added with furan-2-carbaldehyde (5.04 g, 52.5 mmol) and stirred at room temperature for 12 hours. The mixture was added with ethyl acetate (200 ml), washed with water (200 ml, twice), dried over anhydrous magnesium sulfate, and then the filtered filtrate was distilled under reduced pressure. The filtrate was separated using silica gel column chromatography to obtain the following compound (2.00 g, 7.96 mmol).

'H-NMR (400MHz, DMSO-t/s) 59.68(s, 1H), 7.66(m, 2H), 7.73(d, 1H), 7.52(t, 1H), 7.44(t, 1H), 7.32(s, 1H)

Preparation Example 1-2)

The compound (2.00 g, 7.96 mmol) obtained in Preparation Example 1-1) was dissolved in dimethylformamide (40 ml), added with sodium bicarbonate (669 mg, 7.96 mmol) and water (10 ml), and then stirred. The mixture was added with (1,1’- bis(diphenylphosphino)ferrocene)palladium(II) chloride (0.796 mmol, 582.4 mg) and 2-methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaboran-2-yl)isoindoline-l-one (2.61 g, 9.55 mmol), and stirred at 80 to 90°C for 1 hour. When the reaction was completed, the mixture was cooled to room temperature, added with water, and then extracted with ethyl acetate (100 ml). The extracted solution was dried with anhydrous magnesium sulfate and concentrated under reduced pressure to obtain a residue. The residue was separated by silica gel column chromatography to obtain the following compound (1.40 g, 4.41 mmol). ’H-NMR (400MHZ, DMSO-de) 59.69(s, 1H), 8.05(d, 1H), 7.96(d, 2H), 7.94(d, 1H), 7.69(t, 1H), 7.66(d, 1H), 7.61(t, 1H), 7.32(d, 1H), 4.22(brs, 2H), 3.26(s, 3H)

Example

Synthesis of N-methyl-2-(l-((5-(2-(2-methyl-l-oxoisoindolin-4- yl)phenyl)furan-2-yl)methyl)-3-oxopiperazin-2-yl)acetamide (Code name BBC1505, Chemical Formula: C27H28N4O4, Exact Mass: 472.21)

5-(2-(2-methyl-l-oxoisoindolin-4-yl)phenyl)furan-2-carboaldehyde (80 mg, 0.252 mmol) obtained in Preparation Example 1-2) was added to di chloromethane (5 ml), added sequentially with sodium triacetoxyborohydride (80.1 mg, 0.378 mmol), acetic acid (10 pl), and methylamine hydrochloride (25.5 mg, 0.378 mmol), and then stirred at room temperature for 12 hours. When the reaction was completed, the mixture was added with water and extracted with dichloromethane (20 ml). The extracted solution was dried with anhydrous magnesium sulfate and concentrated under reduced pressure to obtain a residue. The residue was separated by silica gel column chromatography to synthesize N-methyl-2-(l-((5-(2-(2-methyl-l- oxoisoindolin-4-yl)phenyl)furan-2-yl)methyl)-3-oxopiperazin-2-yl)acetamide (BBC 1505).

- ’H NMR: 400 MHz, CD3OD 5 (ppm) 8.0 (dd, J = 12, 7.6 Hz, 2H), 7.6 (t, J =

7.6 Hz, 1H), 7.50 (td, J = 7.6, 1.2 Hz, 1H), 7.40-7.47 (m, 2H), 7.32 (d, J = 6.4 Hz, 1H), 6.22 (d, J = 3.2 Hz, 1H), 5.80 (d, J = 3.2 Hz, 1H), 4.08 (br s, 2H), 3.68 (s, 2H), 3.20- 3.26 (m, 2H), 3.11-3.17 (m, 1H), 3.08 (s, 3H), 2.76-2.81 (m, 1H), 2.73 (d, J = 4.8 Hz, 1H), 2.66-2.71 (m, 4H), 2.29 (m, 1H)

- LCMS: m/z = 473.4 (M+H)+, Rt = 1.256 min

Experimental Examples

Experimental Example 1

In order to confirm a therapeutic effect of the composition containing the compound according to the present invention as an effective ingredient on osteoarthritis, an experiment (in vivo) for confirming the therapeutic effect on osteoarthritis was performed using a rodent disease model (DMM).

A 12-week-old B6 mouse was first anesthetized, the mouse knee was resected in a straight line using a No. 11 blade, and then the knee ligament was pushed to the side to confirm and resect the meniscus, and then sutured. Only a mouse knee skin portion of the opposite leg was resected in a straight line, and then the knee ligament was pushed to the side to restore the leg to its original state. When analyzing the mouse knee for 2 to 10 weeks after surgery, it was confirmed that osteoarthritis symptoms appeared from 4 weeks after surgery and cartilage damage was caused after 6 weeks. As described above, osteoarthritis was caused and a BBC 1505 drug was injected into the mouse knee joint from 4 weeks later. Six weeks after the start of substance administration, the mouse knee was fixed with 4% PFA and decalcification was performed for 2 weeks using an EDTA solution. After paraffinization, paraffin was absorbed into the knee cartilage and then Safranin O-staining (see FIG. 3) was measured to confirm the cartilage tissue destruction inhibition ability. In addition, changes in OARSI grade and SBP thickness were measured to confirm the destruction inhibition ability of mouse knee cartilage tissue and the osteoarthritis therapeutic effect

(see FIG. 4).

After inducing osteoarthritis (DMM), a B6 mouse was administered with the BBC 1505 drug, and the bone tissue was confirmed through Safranin O-staining for 6 weeks, and as a result, it was confirmed that an effect of inhibiting cartilage destruction was shown compared to a negative control group. In addition, even in the result of histological evaluation performed based on the OARSI score, it was confirmed that the composition showed an osteoarthritis therapeutic effect according to the composition administration. The thickness of a subchondral bone plate (SBP) was also reduced, thereby confirming the osteoarthritis therapeutic effect of a test substance.

Experimental Example 2

An experiment (in vivo) for confirming the osteoarthritis therapeutic effect was performed using a rabbit disease model (ACLT).

Before inducing osteoarthritis using a male New Zealand white rabbit, the hair around the right knee of the rabbit was widely removed using a clipper. The incision site was widely disinfected using povidone and 70% alcohol, and then the skin of the right knee was incised. The surrounding tissue was bluntly dissected to expose the articular surface of the right femoral distal end. The anterior cruciate ligament was cut with surgical scissors, and a defect wound was fabricated on the medial articular surface, and then the wound was sutured using a stapler. After induction of osteoarthritis, antibiotics (cefazolin) and analgesics (tramadol) were administered for 3 days. One week after induction of cartilage damage, the rabbit was subjected to artificial exercise (30 min/day) once/day for 1 week. At 2 weeks after induction of osteoarthritis, the weight-bearing distribution rate values of animals in an arthritis- induced group were measured and ranked, and randomly distributed, so that the average value of the osteoarthritis-induced group was distributed as evenly as possible. The distributed animals were anesthetized and a BBC 1505 drug, a negative control group (vehicle), and a positive control group (hyaluronic acid) were administered into the right knee joints. On day 84 from the start of test substance administration, the right knee joint was excised and fixed in a 10% neutral buffered formalin solution. The fixed tissue was stained using H&E and Safranin O-staining (see FIG. 5) to confirm cartilage destruction inhibition ability. In addition, immunohistochemical staining (Collagen type 2) was performed, and histopathological changes were observed using an optical microscope according to the OARSI evaluation method (see FIG. 6).

Using the articular tissue extracted at autopsy, OARSI grading was performed through H&E staining and Safranin O-staining. The femur and tibia were each divided into medial and lateral, and the product of Grade and Stage was calculated, and the average of the values was expressed as a value of an individual. In the case of an induced control group, most individuals were observed to have lesions of Grade 3 or higher with fissures appearing on the articular surface, and on Safranin O-staining, it was observed that the proteoglycan content in the cartilage was reduced compared to a normal control group. In addition, all individuals showed average Stage 2 or higher in the range of articular surface damage, and all individuals had areas of Stage 3 or higher. Based on the results, it was confirmed that the degree of arthritis induction in the test model was appropriate. G3 and G5 groups, which were groups administered with the test substance, showed about Grade 2 lesions with articular surface fibrillation on average, so that the OARSI score was observed to be statistically significantly lower than that of the induced control group.

As a result of performing immunohistochemical staining on Type 2 collagen using the extracted articular tissue, the Type 2 collagen positive area levels of the G3 and G5 groups were observed to be statistically significantly higher than that of the induced control group, and it was confirmed that the G3 and G5 groups had excellent cartilage regeneration ability compared to the positive control group. The histopathological examination, immunofluorescence staining result, and ELISA analysis result showed a trend corresponding to the results of measuring the knee circumference and weight load distribution rate, and based on these results, it was confirmed that the BBC 1505 drug was effective in alleviating pain caused by osteoarthritis induced by anterior cruciate ligament transection and meniscus transection and improving indicators related to cartilage regeneration.

Claims

1. A pharmaceutical composition for the prevention or treatment of arthritis, comprising a compound represented by Chemical Formula I below or a pharmaceutically acceptable salt thereof as an active ingredient:

[Chemical Formula I]

2. The pharmaceutical composition for the prevention or treatment of arthritis of claim 1, wherein the compound represented by Chemical Formula I is N-methyl-2-(l- ((5-(2-(2-methyl-l-oxoisoindolin-4-yl)phenyl)furan-2-yl)methyl)-3-oxopiperazin-2- yl)acetamide.

3. The pharmaceutical composition for the prevention or treatment of arthritis of claim 1 or 2, wherein the composition has cartilage tissue regeneration ability.

4. The pharmaceutical composition for the prevention or treatment of arthritis of claim 1 or 2, wherein the composition has cartilage tissue destruction inhibition ability.

5. The pharmaceutical composition for the prevention or treatment of arthritis of claim 1 or 2, wherein the arthritis is at least one selected from the group consisting of osteoarthritis, rheumatoid arthritis, degenerative arthritis, psoriatic arthritis, and reactive arthritis.

6. The pharmaceutical composition for the prevention or treatment of arthritis of claim 1, wherein the compound represented by Chemical Formula I or the pharmaceutically acceptable salt thereof is contained in an amount of 0.00001 to 50 wt% based on the total weight of the composition.

7. A food composition for the prevention or treatment of arthritis, comprising a compound represented by Chemical Formula I below or a pharmaceutically acceptable salt thereof as an active ingredient:

[Chemical Formula I]

8. The food composition for the prevention or treatment of arthritis of claim 7, wherein the arthritis is at least one selected from the group consisting of osteoarthritis, rheumatoid arthritis, degenerative arthritis, psoriatic arthritis, and reactive arthritis.

9. The food composition for the prevention or treatment of arthritis of claim 7, wherein the compound represented by Chemical Formula I or the pharmaceutically acceptable salt thereof is contained in an amount of 0.00001 to 50 wt% based on the total weight of the composition.