KR101987971B1 - Composition for Preventing or Treating Arthritis, Comprising Aromatic Ketone Compound, Melicope pteleifolia Extract, or Fraction Thereof Containing the Same - Google Patents

Abstract

The present invention relates to a composition for alleviating, preventing or treating arthritis, which contains a compound represented by chemical formula 1, a pharmaceutically acceptable salt of the compound, a Melicope pteleifolia extract containing the compound, or a fraction thereof as an active component, wherein the composition is administered to an individual of arthritis, thereby being able to alleviate inflammation, edema, and panus, and to alleviate cartilage loss and bone erosion.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for preventing or treating arthritis, which comprises an aromatic ketone compound, a trihydrate extract thereof or a fraction thereof, and a composition for preventing or treating arthritis,

The present application relates to aromatic ketone-based compounds, pharmaceutically acceptable salts of such compounds, tertiary ( Melicope The present invention also relates to a pharmaceutical composition for preventing or treating arthritis.

Arthritis is caused by inflammation of the joints, especially the knee joints, due to a variety of causes, including edema and fever. Arthritis can be categorized into at least 100 forms, but the most frequent manifestations are osteoarthritis (OA) and rheumatoid arthritis (RA).

Degenerative arthritis is the most common form of arthritis, a degenerative disease that progresses due to progressive wear of articular cartilage and is most common in middle-aged and elderly people. Causes of articular cartilage damage include overweight, joint trauma, peripheral bone disease, muscle weakness, and joint nerve damage. It may also be caused by genetic predisposition. The joints in which degenerative arthritis mainly occurs are the hip, knee, and lumbar spine, and may occur in the fingers or toes. Degenerative arthritis, unlike rheumatoid arthritis, does not occur in the wrists, elbows or ankles.

Rheumatoid arthritis is a systemic inflammatory disease that is a type of arthritis with a prevalence of 1% of the population. It is an inflammatory reaction caused by an autoimmune mechanism, which is a typical refractory disease resulting in severe and sudden joint destruction and corresponding dysfunction. The prevalence of Korean arthritis using the National Health and Nutrition Examination Survey, and the change by year, Korean Journal of Rheumatology, Vol. 15, No. 1, page 11-26, March 2008).

Rheumatoid arthritis is the most serious disease and disorder of arthritis. The mortality rate is known to be similar to that of malignant tumors and cardiovascular diseases in the long term. The non-bacillary inflammatory reaction appears chronically in the synovial membrane, which is characterized by proliferation of synovial membrane and increased amount of synovial fluid resulting in edema and pain of the joint. Rheumatoid arthritis affects not only the joints but also the various organs of the body, so it is sometimes called a rheumatic disease. Key features include activation of the immune response. The precise factors responsible for this response are not yet known, but autoimmunity is known to play an important role in the chronicity and progression of the disease. In other words, rheumatoid arthritis is a persistent autoimmune reaction that causes T cells to play an important role in inflammatory mediators and cytokines, resulting in destruction of joints. The main symptoms that appear are fatigue, helplessness, pain and arthritis And accompanied by fever and weakness.

As a typical treatment for arthritis, Cox-2 inhibitor is mainly used for osteoarthritis, and it is used for reduction of acute pain through reduction of inflammation and improvement of joint function. However, it is effective for direct treatment of rheumatoid arthritis patients and prevention of joint destruction Is difficult to expect.

In addition, the use of non-steroidal anti-inflammatory drugs (NSAIDs) to treat rheumatoid arthritis has been shown to improve joint pain and improve symptoms, And gastrointestinal side effects such as gastric duodenal ulcer may become a problem during long-term use. Steroid Hormone System The medication that controls the inflammation, the face should be rounded, the weight gain, diabetes, hypertension, etc.

The development of an effective and safe arthritis treatment with the side effects and defects is continuing, and it is very important to develop a medicament having a low side effect because it is necessary to take the medicine for a long time to treat rheumatoid arthritis It is an urgent matter.

Under these circumstances, the present inventors have made intensive efforts to develop a preventive or therapeutic agent for arthritis, which is harmless to humans. As a result, the present inventors have found that a composition for preventing or treating arthritis comprising an aromatic ketone compound, a trichome extract or a fraction thereof Has been administered to an individual of arthritis to alleviate inflammation, edema, and panus, and can alleviate cartilage loss and bone erosion, thus completing the present invention.

One object of the present invention is three garage including a pharmaceutically acceptable salt thereof, wherein the compound of the compound, the compound represented by Formula 1 (Melicope The present invention also provides a pharmaceutical composition for preventing or treating arthritis, which comprises, as an active ingredient, an extract or a fraction thereof.

It is another object of the present invention three garage including a pharmaceutically acceptable salt thereof, wherein the compound of the compound, the compound represented by Formula 1 (Melicope pteleifolia ) extract or a fraction thereof as an active ingredient. The present invention also provides a food composition for preventing or ameliorating arthritis.

Another object of the present invention is to provide a compound represented by the formula (I), a pharmaceutically acceptable salt of the compound, a triple- phase ( Melicope pteleifolia ) extract or a fraction thereof as an active ingredient. The present invention also provides a feed composition for preventing or ameliorating arthritis.

Another object of the present invention is to provide a compound represented by the formula (I), a pharmaceutically acceptable salt of the compound, a triple- phase ( Melicope pelleifolia ) extract or a fraction thereof as an active ingredient to an individual other than a human.

Hereinafter, the present invention will be described in more detail.

On the other hand, each description and embodiment disclosed herein can be applied to each other description and embodiment. That is, all combinations of the various elements disclosed herein are within the scope of the present invention. Further, the scope of the present invention can not be said to be limited by the following detailed description.

In addition, those of ordinary skill in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described in this application. Further, such equivalents are intended to be included in the present invention.

According to one aspect of the present invention for achieving the above object, the present invention provides a compound represented by the following formula (1), a pharmaceutically acceptable salt of the compound, a triple heme comprising the compound pteleifolia ) extract or a fraction thereof as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating arthritis.

[Chemical Formula 1]

In the present invention, the "compound represented by the formula (1)" may be used for the prophylactic or therapeutic use of arthritis. The compound is also referred to herein as compound 1. The extract or fraction may be separated from the extract. However, the compound of formula (1) is not limited to the method of obtaining the compound, and may be used for the prevention or treatment of arthritis.

The compound 1 is not limited to a stereoisomer having the structure of Formula 1 and showing the use of arthritis for preventing or treating arthritis. Specifically, a β-D-glucose glycoside may be used as an active ingredient, such as a compound represented by the following formula (2). This is referred to herein as EL6.

(2)

The compound 1 is administered to an individual of arthritis to alleviate inflammation, edema, and panus, and can alleviate cartilage loss and bone erosion. In one embodiment of the present invention, the administration of EL6 to an animal model of arthritis confirmed that inflammation, edema, and panus were alleviated, and further, the degree of cartilage and erosion was alleviated (Example 2).

The compound can be used in the form of a pharmaceutically acceptable salt. The term "pharmaceutically acceptable salt" of the present invention means a concentration that is relatively non-toxic to a subject and has a harmless effective action, wherein the side effects caused by the salt are any all organic or inorganic salts of the compound that do not impair the beneficial efficacy of Compound An inorganic addition salt. In addition, the compound 1 may be used alone or in combination with another pharmaceutically active compound or in a set.

The pharmaceutically acceptable salts of the compound 1 of the present invention refer to salts prepared according to methods customary in the art, and such methods are known to those skilled in the art. In particular, the pharmaceutically acceptable salts include, but are not limited to, salts derived from inorganic and organic acids and bases which are pharmacologically or physiologically acceptable.

The acid addition salt is prepared by a conventional method, for example, by dissolving the compound in an excess amount of an acid aqueous solution, and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. The same molar amount of the compound and the acid or alcohol (e.g., glycol monomethyl ether) in water may be heated and then the mixture may be evaporated to dryness, or the precipitated salt may be subjected to suction filtration. As the free acid, organic acids and inorganic acids can be used. As the inorganic acids, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid and the like can be used. Examples of the organic acids include methanesulfonic acid, p- toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycollic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid and the like can be used , But are not limited to these.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal salt or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess amount of an alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the non-soluble salt salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically acceptable to produce sodium, potassium, or calcium salt, but not limited thereto. The corresponding silver salt can also be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

The present invention includes both the compound 1 or a pharmaceutically acceptable salt thereof as well as a solvate or hydrate thereof exhibiting the same effect, which can be prepared therefrom, within the scope of the present invention.

As used herein, the term " triplet "is a shrub with Rutaceae having the scientific name Melicope pteleifolia . Inhabits forests with mixed evergreen and deciduous forests, at the edge of forests, bushes and marsh forests, about 2300-2800 meters above sea level. Specifically, it lives in Cambodia, Laos, Myanmar, Thailand, Taiwan and Vietnam. In the present invention, the tertiary structure can be obtained by purchasing a commercially sold tertiary structure, or harvested or cultivated in nature.

As used herein, the term "extract" means an extract obtained by extracting tertiary amino acids. The trichotomous extract is prepared by mixing trical and pulverized products in a volume of about 2 to 20 times, preferably about 3 to 5 times the dry weight of water, of ₁ to 4 carbon atoms (C ₁ to 4) such as methanol, ethanol, ₄ ) or a mixed solvent having a mixing ratio of about 1: 0.1 to 1:10 of them is used as an elution solvent, and the extraction temperature is 20 to 100 ° C, preferably room temperature, and the extraction period is about Extraction can be carried out for 1 to 4 days using hot water extraction, cold extraction, reflux cooling extraction, ultrasonic extraction, or the like. However, it is possible to use a substance having the activity of preventing or treating arthritis, specifically, And can be used without limitation. More specifically, it may be a result of several times extraction with water or ethanol for 1 to 3 hours using an ultrasonic extractor, but it is not limited to this, as long as it can exhibit the activity of preventing or treating arthritis. Or a concentrate, a dried product obtained by drying the extract, or any of these adjusted products or purified products. The extract can be extracted from various organs of natural, hybrids, and variant plants and extracted from plant tissue cultures as well as root, top, stem, leaf, fruit body, fruit skin. In one embodiment of the present invention, trilobal leaves were pulverized and then water or ethanol was added thereto, followed by ultrasonic extraction for 2 hours.

In one embodiment of the present invention, the content of EL6 as the compound 1 was the highest at the extraction condition using the extract of 70% ethanol, but the use of the resin to optimize the extraction amount and the fraction containing Compound 1 Considering convenience, it can be extracted using water or other concentrations of ethanol.

The term "fraction " as used herein means a product obtained by a fractionation method for separating a specific component or a specific group from a mixture containing various constituents. The fraction of the present invention is not limited to the method for obtaining a substance used for preventing or treating arthritis, particularly a fraction containing Compound 1. [ Specifically, the tertiary fraction of the present invention is prepared by suspending the above-mentioned crude extract in 1 to 10 times, particularly 5 to 8 times, water, and then subjecting it to various fractionation methods such as adsorption chromatography, ion exchange chromatography, Can be used to obtain fractions.

More specifically, the extract may be fractionated by passing the extract through an ion exchange resin under a solvent flow. In the present invention, the ion exchange resin used for fractionation of the extract is a synthetic adsorbent, Resin (Diaion HP-20 series, SP825 series, AXT204 series, XAD1600T series, MN200 series). The type of the solvent is not particularly limited, and any solvent known in the art can be used. Non-limiting examples of the fraction solvent include polar solvents such as water, alcohol and the like; And non-polar solvents such as hexane, ethylacetate, acetone, and chloroform. Specifically, it is possible to use a lower alcohol solvent having a carbon number of ₁ (C 1) to ₄ (C 4), can be more specifically used for the ethanol solvent of from 5 to 50% (v / v). These may be used alone or in combination of two or more. In a specific example of the present invention, the content of EL6 was the highest in the fraction obtained by using Diaion HP-20 as an adsorbent and using a 40% (v / v) ethanol solvent. In addition, when the fraction of the above example was administered to an animal model of arthritis, it was confirmed that inflammation, edema, and panus were alleviated (Example 2).

As used herein, the term "arthritis" includes the symptoms of inflammation in the joints of an individual and the symptoms accompanying it. The arthritis may be accompanied by pain, edema, panus and / or stiff symptoms.

In the present invention, arthritis refers to all diseases classified as arthritis, and includes all kinds of diseases classified into arthritis, including acute rheumatoid arthritis, chronic rheumatoid arthritis, degenerative arthritis, obturator arthritis, gouty arthritis, atrophic arthritis, chronic inflammatory arthritis, , Arthritis, arthritis, arthritis, arthritis, arthritis, arthritis, arthritis, arthritis, arthritis, arthritis, arthritis, arthritis, arthritis, arthritis and arthritis.

EL6 as an active ingredient of the present invention, EL6, is administered to an individual of arthritis to alleviate inflammation, edema, and panus, and can alleviate cartilage loss and bone erosion (Example 2) It is possible to achieve direct arthritis treatment beyond simply relieving inflammation.

The pharmaceutical compositions herein have the use of "prevention " and / or" treatment "of arthritis. For prophylactic uses, the pharmaceutical compositions herein are administered to individuals suspected of having, or at risk for developing, the diseases, disorders, or conditions described herein. For therapeutic use, the pharmaceutical compositions herein are administered in an amount sufficient to treat or at least partially arrest the symptoms of the disease, disorder or condition described herein in an individual, such as a patient already suffering from the disorder as described herein . The effective amount for such use will depend upon the severity and course of the disease, disorder or condition, previous treatment, the health condition of the individual and responsiveness to the drug, and the judgment of the physician or veterinarian.

It may further comprise suitable carriers, excipients or diluents conventionally used in the manufacture of the pharmaceutical compositions herein. At this time, the content of the compound 1 contained in the composition is not particularly limited, but it may include 0.0001% by weight to 10% by weight, preferably 0.001% by weight to 1% by weight based on the total weight of the composition.

The pharmaceutical composition may be any one selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, solutions, emulsions, syrups, sterilized aqueous solutions, nonaqueous solvents, suspensions, emulsions, And may be oral or parenteral formulations of various forms. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules, and the like, which may contain one or more excipients such as starch, calcium carbonate, sucrose or lactose lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the non-aqueous solvent and the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

The composition of the present invention may be administered to a subject in a pharmaceutically effective amount.

The term "pharmaceutically effective amount" as used herein means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will vary depending on the species and severity, age, sex, The type of drug, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the rate of release, the duration of the treatment, factors including co-administered drugs, and other factors well known in the medical arts. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And can be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without adverse effect, and can be easily determined by those skilled in the art. The preferred dosage of the composition of the present invention depends on the condition and the weight of the patient, the degree of the disease, the type of the drug, the administration route and the period, and the administration may be carried out once a day or divided into several times. The composition is not particularly limited as long as it is an object for treating arthritis, and any composition can be applied. The mode of administration is not limited as long as it is a conventional method in the art. For example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

When the pharmaceutical composition of the present application is administered to the subject, inflammation, edema, and panus are alleviated, and cartilage loss and bone erosion can be alleviated.

In another aspect of the present invention for solving the above-mentioned problems, the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable salt of the compound or a triple extract ( Melicope pteleifolia ) comprising the compound or a fraction thereof as an active ingredient , And a food composition for preventing or ameliorating arthritis.

[Chemical Formula 1]

.

.

The terms used here are the same as those described above.

As used herein, the term "improvement" refers to any activity in which the suspicion of arthritis and symptoms of an invention are alleviated or benefited by using a composition comprising Compound 1 as an active ingredient.

The food composition for preventing or ameliorating arthritis of the present invention includes forms such as pills, powders, granules, infusions, tablets, capsules or liquid preparations. Foods to which the composition of the present invention can be added include, for example, various Foodstuffs, for example, beverages, gums, tea, vitamin complexes, and health supplement foods.

There are no particular restrictions on other ingredients other than those containing Compound 1 as an essential ingredient that can be included in the food composition of the present invention, and may contain various herbal medicine extracts, food supplementary additives, natural carbohydrates, .

In addition, the food-aid additive includes food-aid additives customary in the art, for example, flavoring agents, flavoring agents, coloring agents, fillers, stabilizers and the like.

Examples of such natural carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. In addition to the above, natural flavoring agents (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used as flavoring agents.

In addition to the above, the food composition of the present invention can be used as a food composition containing various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and fillers (cheese, chocolate etc.), pectic acid and its salts, Salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, it may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components may be used independently or in combination.

In the present invention, the health supplement food includes health functional food, health food and the like. The term "functional food" as used herein means the same term as "food for special health use" (FoSHU). In addition to nutrition, It means food. Here, the term "function (surname)" means that the structure and function of the human body have a beneficial effect for health use such as controlling nutrients or physiological action. The food of the present invention can be prepared by a method commonly used in the art and can be prepared by adding raw materials and ingredients which are conventionally added in the art. In addition, the formulations of the food can also be produced without restrictions as long as they are formulations recognized as food. The food composition of the present invention can be manufactured in various forms, and unlike general pharmaceuticals, it has advantages of being free from side effects that may occur when a food is used as a raw material for a long time, and is excellent in portability.

In another aspect of the present invention for solving the above problems, the present invention provides a pharmaceutical composition comprising a compound represented by the formula (1), a pharmaceutically acceptable salt of the compound, or a triple- pteleifolia ) extract or a fraction thereof as an active ingredient. The present invention also provides a feed composition for preventing or ameliorating arthritis.

[Chemical Formula 1]

The terms used here are the same as those described above.

As used herein, the term "feed" means any natural or artificial diet, single meal, or the like ingredients described above for feeding, ingesting, digesting, or suitable for the animal. Specifically, the feed containing the composition for preventing or treating arthritis according to the present invention as an active ingredient can be produced in various types of feed known in the art, and preferably includes a rich feed, a feed and / or a special feed .

Concentrated feeds include seeds containing cereals such as wheat, oats, and corn, and by-products such as rice bran, wheat bran, barley, etc., Which is the by-product of the starch residue, which is the main component of the starch residue, which is the remaining product of starch minerals, sweet potatoes, potatoes, etc., which is a byproduct obtained by concentrating the fresh liquid product obtained from fish meal, fish meal, fish residue, The animal feed such as fish soluble, meat powder, blood meal, wheat meal, skim milk powder, cheese in milk, dried whey in the case of casein production in skim milk, dry yeast, yeast, Chlorella, algae, but not limited thereto.

Roughage includes root vegetables such as wild grasses, grasses and fodder, raw turnip for feed, turnip for feed, feed bit, turnip root, Silage, which is filled with lactic acid and fermented by lactic acid, hay, dried hay, herbage straw, straw of leguminous crops, leaves of leguminous plants, and the like. Special diets include mineral feeds such as fodder, rock salt, urea and its derivatives such as diuretic isobutane, nutrients such as diuretic isobutane, and nutrients that can be supplemented when only natural feed ingredients are mixed or supplemented with diets Feed additives, dietary supplements, which are substances added in small quantities, but are not limited thereto.

The composition for the feed comprising the composition for preventing or treating arthritis according to the present invention may be prepared by mixing the compound represented by the formula (1), the pharmaceutically acceptable salt of the compound , Or a tertiary ( Melicope pteleifolia ) extract or a fraction thereof.

In another aspect of the present invention for solving the above problems, the present invention provides a pharmaceutical composition comprising a compound represented by the formula (1), a pharmaceutically acceptable salt of the compound, or a triple- The present invention provides a method of preventing or treating arthritis, comprising administering to a subject a composition comprising, as an active ingredient, an extract or fraction thereof of pteleifolia .

[Chemical Formula 1]

The terms used here are the same as those described above.

In the present invention, the term "individual" means all animals, including humans, who have arthritis or can develop arthritis. By administering the pharmaceutical composition of the present invention to suspect individuals of arthritis, the individual can be efficiently treated.

As used herein, the term "administering " means introducing the pharmaceutical composition of the present invention to a suspected individual of arthritis by any appropriate method, and the administration route may be administered through various routes of oral or parenteral administration, .

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

As used herein, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and the effective dose level will vary depending on the species and severity, age, sex, , The activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and rate of release, the duration of the treatment, factors including co-administered drugs, and other factors well known in the medical arts. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And can be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without adverse effect, and can be easily determined by those skilled in the art.

The pharmaceutical composition of the present invention is not particularly limited as long as it is an object for prevention or treatment of arthritis, and any of them can be applied. For example, any non-human animal such as a monkey, a dog, a cat, a rabbit, a guinea pig, a rat, a mouse, a cattle, a sheep, a pig or a goat can be used. Subcutaneous, intraperitoneal, intrapulmonary, and intranasal, and may be administered by a suitable method, including localized administration, if necessary, for localized treatment. The preferred dosage of the pharmaceutical composition of the present invention varies depending on the condition and body weight of the individual, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. For example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.

Suitable total daily doses may be determined by the treatment within the scope of sound medical judgment and are generally in the range of 0.001 to 1000 mg / kg, preferably 0.05 to 200 mg / kg, more preferably 0.1 to 100 mg / kg can be administered once or several times a day.

The compositions herein may be administered to an arthritis or suspected individual to alleviate inflammation, edema, and panus, or to alleviate cartilage loss and bone erosion.

1 and 2 are each a view showing 1 H NMR and 13 C NMR spectra of EL6 obtained from the triplicate.
Figures 3 and 4 show the HSQC and HMBC spectra of EL6, respectively, obtained from triplicate.
Figure 5 shows an HPLC chromatogram of EL6 obtained from triplicate.
6 is a schematic diagram showing a drug administration schedule of an arthritic animal model.
FIG. 7 is a diagram showing arthritic index when 100 mg / kg of fractional fraction is administered to an animal model of arthritis.
8 is a graph showing the arthritis index when EL6 25 mg / kg (expressed as EL6 25) and 50 mg / kg (expressed as EL6 50) are administered to an animal model of arthritis.
FIG. 9 shows the thickness of the ankle (FIG. 9A) and the ankle appearance (FIG. 9B) when EL6 25 mg / kg (expressed as EL6 25) and 50 mg / Fig.
10 is a graph showing H & E staining photographs (FIG. 10A) and histopathological arthritis index (FIG. 10B) of joint slices when EL6 50 mg / kg (denoted EL650) .
FIG. 11 is a graph showing the result of microcontrast analysis of bone and Safranin O and Toluidine blue staining of cartilage when EL6 50 mg / kg (expressed as EL6 50) was administered to an animal model of arthritis.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are intended to illustrate the present invention, but the scope of the present invention is not limited thereto.

Example  1: from trichotomous extract Fraction  Preparation and isolation of active compounds

Determination of solvent extraction conditions of active compounds

The triplet used in the present invention was purchased from My Duc herbal distract in Hanoi, Vietnam in 2017 and was used by Tran The Bach of Vietnam Institute of Ecology and Biological Resources, Vietnam . 10 g of the trilobate was treated with 100 ml of distilled water, 30% [v / v] ethanol aqueous solution, 70% [v / v] ethanol aqueous solution, 95% ethanol, methanol and ethyl acetate for 2 hours, And extracted three times. Then, extracts from which the solid content was removed were concentrated under reduced pressure to obtain 0.33 to 2.08 g of extracts per each extract.

The contents of EL6 were evaluated by high performance liquid chromatography / mass spectrometry (HPLC / MS) from these extracts, and the results are shown in Table 1.

Extraction solvent Extraction quantity Content of EL6 in the extract (%) Distilled water 2.08 g 4.38% 30% ethanol 1.55 g 2.86% 50% ethanol 1.19 g 5.60% 70% ethanol 1.59 g 6.07% 95% ethanol 0.62 g 1.45% Methanol 1.12g 5.39% Ethyl acetate 0.33 g 0% (not detected)

As shown in Table 1, the highest content of EL6 was obtained under the condition of using 70% ethanol, but the distilled water was used in consideration of the ease of use of the resin to optimize the extraction amount and the fraction containing EL6 And the extraction method using the ultrasonic wave extractor 3 times was set as the optimum extraction condition.

Truncated fraction Purified water  Produce

The extract was obtained by cutting the three-dimensional cartilage into an extractor, adding 5-8 times of purified water, and then using an ultrasonic extractor for 30 to 50 hours. The extracts extracted three times were combined and filtered to obtain a filtrate.

The distilled water extract solution was sampled and passed through a Diaion HP-20 resin (synthetic adsorbent resin, Mitsubishi Chemical) in an amount of 5-10 times the weight of the raw material. (V / v) ethanol, 60% (v / v) ethanol, 80% (v / v) ethanol and 100% (v / v) ethanol Respectively. Table 2 shows that EL6 adsorbed on Diaion HP-20 was dissolved in distilled water, 40% (v / v) ethanol, 60% (v / v) ethanol, 80% (v / v) ethanol and 100% The degree of elution during use was analyzed by HPLC. The contents of EL6 and acetaphenone compounds were evaluated by high performance liquid chromatography / mass spectrometry (HPLC / MS), and it was confirmed that the optimum solvent was the maximum when 40% (v / v) ethanol was used.

Eluting solvent condition Distilled water 40% EtOH 60% EtOH 80% EtOH 100% EtOH EL6 0.8% 35.5% 0.06% 0% (not detected) 0% (not detected)

Thereafter, the eluate eluted in 40% (v / v) ethanol was filtered, concentrated in a vacuum concentrator, and lyophilized to obtain dry extract.

From trash  The active compound EL6 (3,5- di -C- beta - glucopyranosyl  phloroacetophenone

A portion of the 40% (v / v) ethanol eluate was concentrated and then 40% (v / v) methanol solvent was added under medium pressure liquid chromatography (MPLC, Watcher® Flash Cartridge, 3 x 15 cm; ≪ / RTI > Then, the obtained fractions were analyzed by Gilson HPLC system [Optima Pak C18 Column; 10 × 250 mm, 5 μm particle size, RS Tech, Korea; _{mobile phase MeOH in H 2 Ocontaining0.1%} HCO 2 H (0-50 min: 15-30% MeOH, 51-60 min: 100% MeOH); flow rate 2 mL / min; UV detection at 205 and 254 nm] was performed to obtain the purified main compound EL6.

The physicochemical properties of the obtained compound EL6 are as follows.

The chemical formula: 3,5- di - C - beta - glucopyranosyl phloroacetophenone

Brownish gum.

[[alpha]] _D ; + 56.4 (c 0.3, MeOH).

UV? _Max (MeOH) (log?) (Nm) 234 (3.48), 286 (3.49).

IR (KBr) v _max 3354, 1621, 1447, 1277, 1048 cm < ^-1 >.

ESIMS m / z 475 [MH ₂ O + H] ⁺ , 491 [MH] ^- .

^{1 H NMR (800 MHz, DMSO-} d 6): 9.15 (1H, s, 6-OH), 4.71 (2H, d, J = 10.0 Hz, H-1 ', 1), 3.61 (4H, m, H -6 ', 6), 3.47 ( 2H, t, J = 9.0 Hz, H-2', 2), 3.33 (2H, t, J = 9.0 Hz, H-4 ', 4), 3.26 (4H, m , H-3 ', 3, 5', 5), 2.60 (3H, s, H-8).

^{13 C NMR (200 MHz, DMSO-} d 6): 104.8 (C-1), 161.3 (C-2, 6), 104.0 (C-3, 5), 161.1 (C-4), 203.5 (C-7 ), 32.9 (C-8), 74.6 (C-1 ', 1), 72.1 (C-2', 2), 77.8 (C-5 ', 5), 60.0 (C-6', 6).

The 1 H NMR and 13 C NMR spectra of the resulting EL6 are shown in Figures 1 and 2, respectively, and the HSQC and HMBC spectra are shown in Figures 3 and 4, respectively. The HPLC chromatogram is also shown in Fig.

Example  2: collagen-induced arthritis in animal models Fraction  And the active ingredient EL6 In vivo  (in vivo ) Efficacy verification

Preparation of animal models

An animal model of collagen induced arthritis (CIA) was prepared. Male DBA / 1J mice at 8 weeks of age were used as test animals. The second type collagen (CII) was dissolved in 0.1 M acetic acid solution to a concentration of 2 mg / ml and dialyzed with a phosphate buffered saline to obtain Complete Fred's Adjuvant (CFA, Chondrex) containing M. tuberculosis, And the immunogen was subcutaneously injected (100 μg / 100 μl) into the base of the tail of the mouse by 100 μl per mouse (first injection). Three weeks later, the same CII was mixed with an equal volume of CFA and subcutaneously injected (100 μg / 100 μl) into the base of the tail of the mouse (second injection). After one week, lipopolysaccharide (LPS) was intraperitoneally injected at 40 μg per mouse, and the experiment was terminated after one week.

Administration and evaluation of arthritis efficacy

Kg of EL6 at 25 mg / kg (expressed as EL6 25) at a concentration of 100 mg / kg (triplicate fraction 100) in vehicle (0.5% carboxymethyl cellulose, CMC) ) And 50 mg / kg (expressed as EL650) once a day until the end of the experiment (Fig. 6).

From the second inoculation as a starting point, the severity of joint inflammation was evaluated once a day until the end of the experiment. At this time, arthritis evaluation was used by adding the points scored according to the following scale on the four legs. The score criteria for arthritis evaluation are as follows.

0 point: normal

1 point: Inflammation and edema confined to one toe

2 points: inflammation and edema on two or more toes or slight edema of the foot

3 points: Inflammation and edema of the foot

4 points: Severe inflammation and edema of the foot, or ankylosis of the foot,

At the end of the experiment, the thickness of the ankle joint was measured with a caliper, and photographs of each test group were taken.

After the end of the experiment, the test animals were sacrificed and the hind paws of the mice were fixed with 10% neutral buffer formalin, and the calcite was removed from the bones and embedded in paraffin. Five-μm articular cartilage was prepared and stained with hematoxylin and eosin (H & E) and histopathologically assessed for severity of joint inflammation. At this time, arthritis evaluation was used by averaging the score according to the following scale. The score criteria for arthritis evaluation are as follows.

0 point: normal

1 point: proliferation of synovial membrane infiltrated with slight inflammation and inflammatory cells

2 points: 1 point granulomatous lesions of synovial tissue with findings

3 points: In addition to the findings of 2 points, pannus formation and bone erosion

To confirm the degree of destruction of cartilage, histologic examination was performed with Safranin O and toluidine blue staining. Micro-tomographs were used to confirm the degree of 3-D cartilage and bone erosion. CT) analysis.

result

In the group administered orally at a dose of 100 mg / kg, the arthritic index was significantly decreased from the 10th day after the second injection of CII to the end of the experiment ( p <0.05, Student's t- test ) (Fig. 7).

The group of oral administration of EL6 at a dose of 25 mg / kg showed an increase in the arthritis index similar to that of the vehicle administered group. However, in the group administered orally at a dose of 50 mg / kg of EL6, ( P <0.05, Student's t- test) (Fig. 8).

At the end of the experiment, the thickness of the ankle joint was 2.94 ± 0.16 mm in the vehicle-treated group, and 2.30 ± 0.11 mm in the group treated with EL6 at a dose of 50 mg / kg, ( P <0.05, Student's t- test), the visual acuity of the ankle joint was decreased in the whole joint of the foot joint (Fig. 9).

Histological examination with H & E staining showed that the vehicle was orally administered with clear synovial proliferation, moderate inflammatory cell infiltration, and intussusception, whereas EL6 was administered orally at a dose of 50 mg / kg vehicle, the quantitative histopathological arthritis index was 1.69 ± 0.25 in the vehicle-treated group and 0.82 ± 0.24 in the group treated with EL6 at the dose of 50 mg / kg EL6 was orally administered at a dose of 50 mg / kg ( p < 0.05, Student's t- test) (Fig. 10).

In the group treated with Vehicle, cartilage loss was observed in Safranin O and Toluidine blue staining of cartilage, and micro-CT analysis showed many bone erosion. On the other hand, in the case of oral administration of EL6 at a dose of 50 mg / kg, such cartilage destruction and bone erosion were clearly recovered (FIG. 11). This shows that Compound 1 is administered to an individual of arthritis to alleviate inflammation, edema, and panus, and to alleviate cartilage loss and bone erosion.

From the above description, it will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention without departing from the scope of the present invention as defined by the appended claims.

Claims (11)

A pharmaceutical composition for preventing or treating arthritis, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt of said compound as an active ingredient:
[Chemical Formula 1]

. 2. The pharmaceutical composition according to claim 1, wherein the compound is obtained from a Melicope pteleifolia extract or a fraction thereof. The pharmaceutical composition according to claim 2, wherein the extract is water, a C ₁ to C ₄ alcohol or a mixed solvent thereof. 3. The pharmaceutical composition according to claim 2, wherein the fraction is eluted with 5-50% (v / v) ethanol in the ion exchange resin. 3. The method of claim 1 or 2, wherein the arthritis is selected from the group consisting of acute rheumatoid arthritis, chronic rheumatoid arthritis, degenerative arthritis, obsessive-compulsive arthritis, gouty arthritis, atrophic arthritis, chronic inflammatory arthritis, infectious arthritis, menopausal arthritis, Arthritis, hypertrophic arthritis, or pyogenic arthritis. ( Melicope pteleifolia ) fraction containing a compound represented by the following formula (1) as an active ingredient. The fraction is obtained by eluting the extract from the ion exchange resin with 5 to 50% (v / v) ethanol Pharmacological composition for the prevention or treatment of arthritis,
[Chemical Formula 1]

. A food composition for preventing or improving arthritis, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient:
[Chemical Formula 1]

. 8. A food composition according to claim 7, wherein the compound is obtained from a Melicope pteleifolia extract or a fraction thereof. ( Melicope pteleifolia ) fraction containing a compound represented by the following formula (1) as an active ingredient. The fraction is obtained by eluting the extract from the ion exchange resin with 5 to 50% (v / v) ethanol Food composition for the prevention or amelioration of arthritis:
[Chemical Formula 1]

. A feed composition for preventing or ameliorating arthritis, comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient:
[Chemical Formula 1]

. A method for preventing or treating arthritis, comprising administering to a subject other than a human a composition comprising an effective ingredient of a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
[Chemical Formula 1]

.

Images