WO2018124695A1 - Nouveau composé flavonoïde séparé de l'extrait de feuilles de stauntonia hexaphylla et composé pour favoriser l'anti-inflammation, formation osseuse ou formation de cartilage ayant la même fonction qu'un principe actif - Google Patents

Nouveau composé flavonoïde séparé de l'extrait de feuilles de stauntonia hexaphylla et composé pour favoriser l'anti-inflammation, formation osseuse ou formation de cartilage ayant la même fonction qu'un principe actif Download PDF

Info

Publication number
WO2018124695A1
WO2018124695A1 PCT/KR2017/015479 KR2017015479W WO2018124695A1 WO 2018124695 A1 WO2018124695 A1 WO 2018124695A1 KR 2017015479 W KR2017015479 W KR 2017015479W WO 2018124695 A1 WO2018124695 A1 WO 2018124695A1
Authority
WO
WIPO (PCT)
Prior art keywords
formula
composition
bone
arthritis
flavonoid compound
Prior art date
Application number
PCT/KR2017/015479
Other languages
English (en)
Korean (ko)
Inventor
이용남
이동구
유지석
최철웅
반상오
강후원
김재용
이규옥
김재갑
Original Assignee
영진약품 주식회사
재단법인 전남생물산업진흥원
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 영진약품 주식회사, 재단법인 전남생물산업진흥원 filed Critical 영진약품 주식회사
Publication of WO2018124695A1 publication Critical patent/WO2018124695A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
    • C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
    • C07H17/06—Benzopyran radicals
    • C07H17/065—Benzo[b]pyrans
    • C07H17/07—Benzo[b]pyran-4-ones
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/306—Foods, ingredients or supplements having a functional effect on health having an effect on bone mass, e.g. osteoporosis prevention
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00—Food ingredients
    • A23V2250/20—Natural extracts
    • A23V2250/21—Plant extracts
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00—Food ingredients
    • A23V2250/20—Natural extracts
    • A23V2250/21—Plant extracts
    • A23V2250/2116—Flavonoids, isoflavones

Definitions

  • the present invention relates to a novel flavonoid compound isolated from honey leaf extract and a composition for promoting anti-inflammatory, bone tissue production or cartilage tissue production comprising the same as an active ingredient, more specifically from dried honey (Stauntoni a hexaphyl la) leaf Extracted with an organic solvent including water, methanol, ethanol, butanol, etc., a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, and then fractionated to obtain the fractionol, which is then purified by chromatography or the like.
  • the present invention relates to a method for preparing a flavonoid compound and a composition for promoting anti-inflammatory or bone tissue production or cartilage tissue production of a novel flavonoid compound prepared by the above method.
  • Bone tissue is generally composed of dense bone with a strong surface of the bone, and the center or both ends of the long bone ( ⁇ ⁇ ) is composed of spongy bone, which is associated with the bone like a mesh. Most canes initially develop as cartilage in connective tissue, which turns into bone tissue in leprosy, some of which are made directly from connective tissue. In the epiphysis, the joint surface is in contact with the neighboring bone, and the surface is covered with articular cartilage, which is a faux bone. Sea sponge shochu in the sponge is characterized by a certain arrangement. The broad course of the bony stem is continuous with the small lumen of sponge shochu and all are filled with bone marrow.
  • Hematopoietic bone marrow has a lot of blood vessels distributed in red color, called red bone marrow.
  • red bone marrow In the structure of the bone, dense and spongy cortices of 5-12 thick layers overlap. In dense form, several layers of concentric layers (harbour layer) are arranged in various directions. There is a bus duct through which blood vessels pass.
  • Bone cells are arranged between the lamellar plates, and are connected to other bone cells adjacent to each other by a protoplasma that leads to an irregular star shape. It has a tough connective tissue periosteum, and nerves and blood vessels are distributed to protect and nourish bone. Defects of the periosteum make it difficult to survive, develop, and regenerate bones.
  • the bone components are 20% moisture, 35% organic matter including cells, 45% minerals, because the constant elasticity of bone is organic. As age increases, minerals (mostly phosphate) increase, resulting in an increase in bone hardness.
  • Osteoclasts produce a variety of enzymes to break down bone, oxidative phosphorylation enzyme TRAPCtartrate-resistant acid phosphatase, which oxidizes and breaks down bone, and matrix metallopeptidase-9, which breaks down the supportive tissue of bone and cartilage.
  • Cathepsin K which breaks down collagen, is known as the main enzyme, and it is used as a special marker for mature osteoclasts.
  • RANKL was treated on macrophages, precursors of osteoclasts, NF- ⁇ and MAPK kinase were activated to produce inflammatory mediators.
  • the C0X enzyme is a major enzyme involved in the biosynthesis of prostaglandin present in vivo (Smith et al., J. Biol. Chem., 271, 33157 (1996)), and two isomeric enzymes, C0X-1 and C0X. -2 is known to exist.
  • the C0X-1 is constantly present in tissues such as the stomach and kidneys, and is involved in maintaining normal homeostasis, while the C0X-2 is involved in mitogen or cytokines during inflammation and other immune reactions. Is an enzyme that is transiently and rapidly expressed in cells.
  • Nitric oxide is produced from L-arginine by NO synthase (N0S) and is produced in many cell types by substances such as endotoxins or cytokines such as UV or external stresses such as UV. do.
  • NO Nitric oxide
  • N0S NO synthase
  • cytokines such as UV or external stresses such as UV. do.
  • Such inflammatory stimuli can increase the expression of inducible NOS (iNOS) in the cell, thereby inducing NO production in the cell, thereby inducing an inflammatory response by activating macrophages. Therefore, in order to effectively alleviate inflammation, studies on substances capable of inhibiting NO production have been conducted.
  • H2 inhibitors H2-B l ockers
  • H2 receptor second histamine receptor
  • H2 inhibitors interfere with the metabolism of other drugs in the liver (potent i n bi bi tors of P-450), so take caution when taken with other drugs and have anti-androgen (Ant i— Androgen) effects in men. Side effects such as gynecomast ia, impotence and decreased libido may occur. In addition, because it crosses the placenta and cerebrovascular barrier, side effects may be more dangerous for pregnant women and the elderly, and may cause headaches, complications, sores, and dizziness. Therefore, natural substance-derived substances can effectively inhibit the production of NO, i NOS and TNF- ⁇ expression, and can effectively inhibit the activity of C0X-2 enzyme. And development of substances that promote osteoblast differentiation, inhibit osteoclast differentiation, and produce bone or cartilage tissue, as well as materials derived from natural substances with little or no risk of side effects or cytotoxicity, and thus limit the amount of their use. This is required.
  • S honey (S untoni a hexaphy lla) is an evergreen vine plant that grows up to 15 m tall in the southern coastal area, along with native creeping vines. In mid-white flowers bloom, and in autumn, dark red fruits of the size of a child's fist hang.
  • the fruit is known as a delicious fruit, as the name is derived from the meaning of 'sweet as honey 1 ', but the fruit is not commercialized due to the coarse seeds and the small flesh.
  • the leaves can be wintered in the southern region, the flesh is thick, and the ornamental value is high. As it is used for garden and pergola, the use is increasing, and accordingly, seedling production is increasing.
  • Patent Document 1 Korean Patent Publication No. 1992-0005995
  • Patent Document 2 Republic of Korea Patent Publication No. 1994-0006596
  • Patent Document 3 Korean Patent Publication No. 0465778
  • Patent Document 4 Korean Patent Publication No. 2004-0108265
  • Patent Document 5 Korean Patent Publication No. 0514916
  • Patent Document 6 Korean Patent Publication No. 2005-0074753
  • Patent Document 8 Korean Patent Publication No. 10-0930927
  • an object of the present invention is to provide an anti-inflammatory, bone tissue or cartilage tissue-promoting pharmaceutical composition comprising a novel active ingredient derived from natural honey.
  • Another object of the present invention to provide a pharmaceutical composition for the prevention or treatment of diseases that destroy bone tissue or cartilage tissue comprising a novel active ingredient derived from the honey.
  • Still another object of the present invention is to provide a food composition for preventing or ameliorating a disease that destroys bone tissue or cartilage tissue including a novel active ingredient derived from the honey.
  • the present invention provides a novel flavonoid compound of Formula 1, an isomer thereof, a pharmaceutically acceptable salt, or solvate isolated from the honey extract.
  • the compounds of the present invention are novel flavonoid compounds that have been isolated and purified for the first time in nature by the present inventors, and they exhibit excellent anti-inflammatory, osteogenic or cartilage tissue-promoting effects. It is very useful as a prophylactic, ameliorating or therapeutic agent for various diseases such as rheumatoid arthritis associated with destruction.
  • Figure 5 is a flavonoid compound HOMO COSY spectrum analysis of the present invention isolated from honey bees:
  • Figure 7 is a flavonoid compound of the formula 2 of the present invention isolated from honey honeycomb BC spectroscopic analysis:
  • FIG. 11 is a C-NMR spectrum analysis result of the full-lavonoid compound 13 of Formula 3 of the present invention isolated from honey bees:
  • FIG. 13 is a HMQC spectrum analysis result of a flavonoid compound of Formula 3 of the present invention isolated from honey bees:
  • FIG. 16 is a graph showing the effect of the flavonoid compound of Formula 1 of the present invention isolated from honeybee on TRAP activity against osteoclasts; 17 is a graph showing the osteoclast differentiation staining results of the flavonoid compound of formula 1 of the present invention isolated from honey;
  • 21 is a graph showing the results of ALP activity staining for osteoblasts (MG— 63) of formula 3 derived from the honey leaf extract:
  • FIG. 22 is a graph showing the effect on the expression of co l agen type I I and aggrecan on chondrocyte differentiation (generation) of the flavonoid compound of Formula 1 of the present invention isolated from honey.
  • the compound of formula 1 is a compound of formula 2 having the following structure (7-(((2,3,4,5,6) -4,5- ⁇ 1 ⁇ 0 -61 1 ⁇ 3 -(((2,3,4,5) -3,4,5-1 ⁇ 11 ydroxyt etr ahydro-2H-pyr an-2-y 1) oxy) tetr ahydr o ⁇ 2H ⁇ pyr an-2-y 1) oxy) ⁇ 2 ⁇ (3, 4-di hydr oxypheny 1) -5-hydr oxy-4H-chromen-4-one) or a compound of formula 3 (7-(((2,3,4,5, 6) —4,5- ⁇ ⁇ ( «-6 ⁇ 11 1-3-(((2,3,4,5) ⁇ 3,4,5- ⁇ ydroxytetrahydro
  • the compounds of Formulas 1 to 3 are provided by the present inventors for the first time.
  • the present invention is a salt of the flavonoid compound represented by any one of formulas (1) to (3), preferably a pharmaceutically acceptable salt to provide .
  • the ⁇ pharmaceutically acceptable salts '' refer to salts that are suitable for use in contact with tissues of humans and animals without causing excessive toxicity, irritation, allergic reactions, etc. within the scope of pure medical judgment. Salts acceptable as are well known in the art, for example, described in detail in S. M. Berge et al., J. Parmaceut i cal Scences, 66, 1, 1977.
  • Suitable base addition salt forms are, for example, ammonium salts, lithium, sodium, potassium, magnesium Alkali salts such as salts, such as calcium and alkaline earth metal salts, salts with organic bases, for example primary, secondary and tertiary aliphatic and aromatic amines, for example methylamine, ethylamine, propylamine , Isopropylamine, four butylamine isomers, dimethylamine, diethylamine, diethan to amine, dipropylamine, diisopropylamine, di-n-butylamine, pyridine, piperidine, morpholine, Trimethylamine, triethylamine, tripropylamine, quinuclidin, pyridine, quinoline and isoquinoline, benzatin, N-methyl-D-glucamine, 2—amino-2— (hydroxymethyl) -1,
  • the present invention also relates to hydrates of the flavonoid compounds represented by any one of Formulas (1) to (3) or their respective chains (si de chain). It may include derivative compounds such as glycosides to which a compound such as gl ucose is bound, etc.
  • the flavonoid compound according to the present invention may be isolated from nature or may be prepared by chemical synthesis of flavonoid compounds known in the art.
  • Can Preferably polar flavonoids compounds of the present invention may be isolated and purified from a natural plant. That is, using all manner of extracting the conventional materials, and separation can be obtained from a portion of the plant or plant.
  • the raw plant (medicinal herb) for producing the flavonoid compound of any one of the formulas (1) to (3) of the present invention is honey, which may use the whole plant or a part of the plant, and a preferred usable site is a leaf.
  • these raw plants In order to obtain the desired extract, it may be appropriately dried and macerated, or only dried to use an appropriate solvent, for example water such as purified water, an organic solvent having 1 to 4 carbon atoms.
  • the organic solvent is not limited to, for example, methanol, ethanol, propanol, isopropanol, butanol, etc.
  • Extraction methods used in the present invention can be used without limitation, all methods commonly used for the extraction of plants or herbal medicines, for example, may be steaming, room temperature extraction, hot water extraction, ultrasonic extraction, percolation method or reflux cooling extraction method. Or not limited thereto.
  • the extraction process may be single, but may be repeated two or more times as necessary.
  • the desired extract may be further subjected to a process such as fractionation for the separation of any one of the compounds of Formulas 1 to 3 of the present invention
  • the fractionation solvent used may be used without limitation the extraction solvent.
  • Purification may also be carried out using purification methods known to those skilled in the art. Such purification methods include, for example, reverse phase part it ion chromatography, normal phase adsorpt i on chromatography, and ion exchange chromatography.
  • the separation and purification may be carried out by concentration gradient chromatography, which is carried out by size exclusion chromatography (Si ze exc l us i on chromatography) or an additional purification method comprising one or more combinations thereof.
  • the present invention According to any of formulas 1-3
  • One flavonoid compound was prepared by extracting water from honey leaves with water, separating and purifying it. Isomers, pharmaceutically acceptable salts or solvates thereof of any of the flavonoid compounds of Formulas 1 to 3 increase the activity of ALP to increase the differentiation and activity of osteoblasts, inhibit TRAP activity of osteoclasts, and osteoclasts.
  • the present invention comprises a flavonoid compound of any one of formulas 1 to 3, isomers, pharmaceutically acceptable salts or solvates thereof as an active ingredient, for promoting anti-inflammatory, bone tissue production or cartilage tissue production It relates to a pharmaceutical composition.
  • Flavonoid compounds of any one of formulas (1) to (3) as an active ingredient in the composition according to the present invention can be appropriately adjusted according to the use form and purpose, patient condition, type of symptom and weight, and 0.000001 to 50 weight based on the total weight of solids 3 ⁇ 4, preferably 0.0001 to 40% by weight. However, this may be increased or decreased depending on the needs of the doser, and may be appropriately increased or decreased depending on various factors such as dietary status, nutritional status, degree of damage of bone tissue or cartilage tissue, and the like.
  • composition according to the invention can be administered to a mammal, including humans, by various routes.
  • the mode of administration may be any of the routinely used methods, and may be administered by, for example, oral or parenteral (for example, skin, vein, intramuscular, subcutaneous), and preferably orally. have.
  • the composition of the present invention is a capsule, such as tablets, soft or hard capsules (CAPSULES), granules (GRANULES), pills (PILLS), limonadese (LEMONADES), aromatherapy (AROMATIC WATERS), respectively, according to a conventional method.
  • Powder POWDERS
  • SYRUPS DECOCTIONS
  • INFUSIONS LIQUIDS AND SOLUTIONS
  • ELIXIRS FLUIDEXTRACTS
  • EMULS IONS SUSPES I ONS Oral formulations such as TROCHES, TINCTURES, SPIRITS, or transdermal, PLASTERS, lotion OPTIONS, LINIMENTS, OPHTALMIC OINTMENTS , Aerosol (AEROSOLS), Ointment (OINTMENTS), Emulsion (EMULS IONS), Suspension (SUSPES IONS), Eye drops (OPHTHALM IC SOLUT IONS), Suppository (SUPPOSITIORIES), INJECTIONS, Catalytics (CATAPLSMA), It may be formulated into a parenteral formulation in the form of a cream (CREAMS), pasta (PASTES) and the like.
  • CREAMS cream
  • PASTES pasta
  • CATAPLSMA Catalytics
  • composition of the present invention may further contain an adjuvant such as a pharmaceutically suitable and physiologically acceptable carrier, excipient and diluent in addition to the combination extract.
  • an adjuvant such as a pharmaceutically suitable and physiologically acceptable carrier, excipient and diluent in addition to the combination extract.
  • an excipient a binder, a disintegrant, a lubricant, a solubilizer, a suspending agent, a preservative or an extender may be formulated.
  • the dosage of the pharmaceutical composition of the present invention may be determined by a specialist according to various factors such as the patient's condition, age, weight, degree of cartilage damage, disease progression, and the like. In addition, it is intended to contain a daily dose or 1/2, 1/3 or 1/4 dose thereof of the pharmaceutical composition per unit dosage form, and may be administered 1 to 6 times a day.
  • the present invention comprises a flavonoid compound of any one of Formulas 1 to 3, its isomers, pharmaceutically acceptable salts or solvates as an active ingredient, the prevention or treatment of diseases that destroy bone tissue or cartilage tissue It relates to a pharmaceutical composition.
  • the disease that destroys or damages the bone or cartilage tissue is not limited thereto, arthritis, destruction of the joint due to autoimmune diseases, multiple myositis, ankylosing spondyl itis, systemic lupus erythematosus, multiple myositis, Polymyalgi a rheumat i ca, osteoporosis, bone metastasis or Paget's di sease.
  • arthritis various forms of arthritis, for example osteoarthritis, rheumatoid arthritis, It may include, but is not limited to, diseases such as degenerative arthritis, psoriatic arthritis, and reactive arthritis. Preferably rheumatoid arthritis.
  • the present invention is to promote the production of bone tissue or cartilage or to destroy bone tissue or cartilage tissue, including flavonoid compounds of any one of Formulas 1 to 3, isomers, pharmaceutically acceptable salts or solvates thereof It relates to a food composition for improving the disease to be damaged.
  • the food is not limited to, but not limited to, health supplements, health functional foods, functional foods, and the like, natural foods, processed foods, general food materials, etc.
  • Flavonoid compounds of formula 1, isomers, pharmaceutically acceptable salts or solvents of the present invention It also includes adding a cargo.
  • the food composition of the present invention may be added as it is or used with other food or food compositions, and may be suitably used according to a conventional method.
  • the mixed amount of the active ingredient can be suitably determined according to the purpose of use thereof.
  • the flavonoid compound of any one of Formulas 1 to 3 of the present invention, isomers, pharmaceutically acceptable salts or solvates thereof is from 0.000001 to the total weight of the raw material of the food or beverage in the manufacture of the food or beverage 50% by weight may be added.
  • an effective dose of the flavonoid compound of any one of Formulas 1 to 3, an isomer thereof, a pharmaceutically acceptable salt or solvate thereof may be used in accordance with the effective dose of the pharmaceutical composition, but may be used for anti-inflammatory, bone or cartilage tissue damage.
  • the active ingredient may be used in an amount above the above range because there is no problem in terms of safety.
  • the food composition may contain organic acids, phosphates, antioxidants, lactose caseins, dextrins, glucose, sugars and sorbobis.
  • the food composition comprising the flavonoid compound of any one of Formulas 1 to 3, an isomer thereof, a pharmaceutically acceptable salt or solvate thereof may be in the form of a beverage such as tablets, hard or soft capsules, granules, powders, solutions, suspensions, and the like.
  • these preparations may be in the form of acceptable conventional carriers, e.g., for oral preparations, excipients, binders, disintegrants, lubricants, solubilizers, suspending agents, preservatives or It can be prepared using an extender or the like.
  • Examples of the food to which the flavonoid compound of any one of Formulas 1 to 3, an isomer thereof, a pharmaceutically acceptable salt or a solvate may be added include meat, sausage bread, chocolate, candy, snacks, confectionary, pizza, and ramen noodles. Other noodles, gums, dairy products including ice cream, various soups, beverages, teas, drinks, alcoholic beverages and vitamin complexes, and other nutritional supplements, but are not limited to these types of food.
  • the UV spectrum in methanol showed maximum absorption at 268, 293 and 394 nm.
  • the -NMR spectrum showed five olefinic (S 6.36, 6.77, 6.79, 6.95, and 7.96), two anmeric (55.00 and 5.20) and one -methyl proton signal ( ⁇ 1.23).
  • the arrangement of the anomeric positions was determined to be in ⁇ form.
  • the chemical shifts of the two anomeric carbons are indicated at ⁇ 100.5 and 105.3.
  • Example 3 Measurement of TRAP activity of osteoclasts by the compound of formula 2 or 3 derived from honey leaf extract
  • Osteoclasts have tartrate—resistant acid phosphatase (TRAP), an acid phosphatase that is characteristically resistant to tartrate, and is used as a cytochemical marker of osteoclasts to distinguish them from other bone tissue cells.
  • TRAP tartrate—resistant acid phosphatase
  • the resulting TRAP activity of osteoclasts by of "(2), three compounds from meolkkul leaf extract of the present invention was tested as follows. As described in Example 2 above,
  • TRAP activity was measured using a TRAP acivity assay kit (AK04F, COSMO BIO CO., LTD). Specifically, after 4 days of incubation, 30 ul / well of the culture supernatant was dispensed on a new 96 plate and the chromogenic properties of the prepared kit. Add 170ul / well of substrate / Tart rate-containing buffer
  • Absorbance is measured at 540 Hz after reaction at 37 ° C. for 30 minutes to 3 hours.
  • TRAP activity expressed the absorbance of the sample as a percentage of the absorbance of the control.
  • the cells were inoculated to 5xl0 5 cells / well in 48—well and treated with differentiation factors (RANKL, MCSF) and the compounds of Formulas 2 and 3 of the present invention and incubated for 4 days. Remove the medium, wash with PBS, and add the substrate solution (1.36 mg / mL 4-nitrophenyl phosphate di sodium salt, 10 mM tartrate, 50 mM citrate buffer pH 4.6) to the cell fixed with 10% formaldehyde.
  • RNKL differentiation factors
  • N, N-dimethylformamide 100 JL was added to prepare a staining solution, and the cells were fixed with 10% formaldehyde.
  • the staining solution was 45 (divided by jL and stained at 37 0 C for 30 minutes and observed under a microscope. It is shown in 17.
  • Example 5 Determination of ALP (alkaline phosphatase) activity of osteoblasts by the compound of formula 3 derived from honey leaf extract
  • osteoblasts exhibit ALP activity
  • the effect of the compound of formula 3 derived from the honey leaf extract according to the present invention on the ALP activity in osteoblasts was measured.
  • the osteoblasts, C2C12 celKATCC® CRL-1772) and MG-63 celKATCC® CRL- 1427) were respectively divided into 48wel l 5 x 10 4 cells / wel 1, respectively.
  • Cells were incubated for 3 days in ⁇ - ⁇ essential medium containing 10% FBS, 0.1% p / s of cell growth medium. Three days later, induction of osteoblast differentiation was performed with a—MEM essential medium containing 1% horse serum and 0.1% p / s.
  • Compounds of formula 3 derived from honey leaf extract obtained in Example 1 were treated with osteoblasts C2C12 cells, MG—63 cells for each concentration (E, 5, 5, and 10ug / ml), and cultured for 5 days. Subsequently, the cells were treated with an AP assay buffer (kit) containing 0.01% Triton X, and centrifuged at 1000 xg for 5 minutes to obtain a sample for ALP activity detection.
  • kit AP assay buffer
  • ALP is used to degrade ALP activity using a change in absorbance at 405 nm with the degradation of p-nitrophenylphosphate (pn itr opheny 1 pho spha te) into p-nitrophenol and phosphate. Measured. Protein concentration was determined using the BioRad protein assay kit. ALP activity was expressed as PNP LiM / min / mg protein.
  • osteoblast differentiation was measured by ALP activity site staining using NBT / BCIP substrate to determine osteoblast differentiation of C2C12 cells and MG— 63 cells. Measured. Specifically, the culture solution of the cells that have been incubated for 5 days is removed, and the cells are washed three times with lxPBS. At room temperature with 10% formalin solution. Fix the cells for 15 minutes and wash the cells 3 times with lxPBS to remove residual formalin. The cells were washed again with lx alkaline phosphaste solution and stained with ALP active sites with NBT / BCIP substrate solution.
  • FIGS. 18 and 19 The results of ALP activity and ALP activity site staining using C2C12 cells are shown graphically in FIGS. 18 and 19, and the results of ALP activity and ALP activity site staining using MG—63 cells are shown graphically in FIGS. 20 and 21, respectively.
  • Test results osteoblasts C2C12 cells, as shown in Figure 18, in the experimental group treated with the compound of formula 3 derived from the honey leaf extract (0.5, 5, 10ug / ml), normal group not treated with BMP-2 It was confirmed that the ALP activity is increased compared to. Specifically, in the positive control group treated with BMP-2 (50 ng / ml), An increase in ALP activity (pNP 72.4 uM; 313.4%) was observed as compared to normal.
  • ALP is an enzyme released by osteoblast differentiation and increased activity. Since the increase of ALP activity is directly related to osteoblast activity and increase, the ALP activity by the compound of formula 3 derived from the honey leaf extract of the present invention is increased. Is to demonstrate the effect of promoting osteoblast formation by direct osteoblast differentiation and increased activity.
  • Example 6 Chondrocyte Differentiation (Generation) by the Compounds of Formulas 2 and 3 Derived from Honey Leaf Extract Biomarker Agrecan, Collagen Type 11 (collagen II) mRNA expression (production) measurement
  • chondrocyte primary cells were used to obtain chondrocyte primary cells. Specifically, 1cm above and below the knee of 5 days old rats are cut and the knee cartilage is stored in a 50ml conical tube containing IX PBS (containing 20% P / S). After washing three times for 15 minutes at room temperature with IX PBS (containing 20% P / S), add 400ul of 1% Collagenase D (or H), 1.2 ml of Trypsin 400, serum free media (DMEM F-12) and add 37 0 C. Incubated for 1 hour with stirring.
  • IX PBS containing 20% P / S
  • DMEM F-12 serum free media
  • the cultured cells were lysed with GIT so kit ion (easy BLUE Total RNA extraction kit, Intron Biotechnology Co., Ltd.), centrifuged at 10,000 rpm for 5 minutes at room temperature, The supernatant was removed to obtain a pellet, 1 ml of 0.1% DEPC solution (Sigma) was added to the pellet, and centrifuged again at 12,000 rpm for 2 minutes to remove the supernatant, and the pellet was obtained. In the obtained pellet, guanidinium 0.5 ml By addition and vortexing.
  • GIT so kit ion Easy BLUE Total RNA extraction kit, Intron Biotechnology Co., Ltd.
  • a phenol / chloroform / iso-amylalchol mixed solution 25: 24: 1 was added, vortexed, and centrifuged at 12,000 rpm for 3 minutes to obtain a supernatant.
  • isopropyl alcohol iso-propylalcol
  • the mixture was left at -20 ° C for 30 minutes. Thereafter, the supernatant was removed by centrifugation at 12,000 rpm for 10 minutes, and then the pellet was washed with 70% aqueous ethanol solution and dried under vacuum to separate RA.
  • the isolated RNA was dissolved in 1 ml of 0.1% DEPC solution and used to measure the mRM content of collagen II (Col 11) and agrecan, which are chondrocyte markers, and GAPDH as a house keeping gene. Used.
  • the real-time PCR results are shown in FIG. 22 when the photographed results were compared with the GAPDH content.
  • the negative control group treated with IL—ipa0ng / ml The amount of m RNA expression of collagen II (Col ⁇ ) and agrecan ( a g Titanan ), which are biomarkers related to chondrocytes, was significantly reduced compared to the normal group not treated with IL- ⁇ .
  • the positive control group treated with BMP-2 100 ng / ml was collagen II (Col), a biomarker associated with chondrocytes.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Mycology (AREA)
  • Biotechnology (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Botany (AREA)
  • Nutrition Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Microbiology (AREA)
  • Medical Informatics (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne un nouveau composé flavonoïde séparé d'une feuille de Stauntonia hexaphylla et une composition pharmaceutique ou alimentaire, pour favoriser la formation osseuse ou la formation de cartilage pour prévenir, remédier ou traiter une destruction osseuse ou cartilagineuse telle que la polyarthrite rhumatoïde, ayant la même fonction qu'un principe actif. La présente invention concerne particulièrement, une composition pharmaceutique ou alimentaire comprenant en tant que principe actif un composé représenté par la formule chimique (1) ayant un effet favorisant la formation osseuse ou cartilagineuse ou un effet de prévention, de correction ou de traitement de la destruction osseuse ou cartilagineuse au moyen de l'inhibition de la différenciation des ostéoclastes et de promotion de l'expression et de la production du collagène de type II et de l'aggrécane.
PCT/KR2017/015479 2016-12-29 2017-12-26 Nouveau composé flavonoïde séparé de l'extrait de feuilles de stauntonia hexaphylla et composé pour favoriser l'anti-inflammation, formation osseuse ou formation de cartilage ayant la même fonction qu'un principe actif WO2018124695A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020160182923A KR102038107B1 (ko) 2016-12-29 2016-12-29 멀꿀 잎 추출물로부터 분리된 신규한 플라보노이드 화합물 및 이를 유효성분으로 하는 항염, 골조직 생성 또는 연골조직 생성 촉진용 조성물
KR10-2016-0182923 2016-12-29

Publications (1)

Publication Number Publication Date
WO2018124695A1 true WO2018124695A1 (fr) 2018-07-05

Family

ID=62710465

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2017/015479 WO2018124695A1 (fr) 2016-12-29 2017-12-26 Nouveau composé flavonoïde séparé de l'extrait de feuilles de stauntonia hexaphylla et composé pour favoriser l'anti-inflammation, formation osseuse ou formation de cartilage ayant la même fonction qu'un principe actif

Country Status (2)

Country Link
KR (1) KR102038107B1 (fr)
WO (1) WO2018124695A1 (fr)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102126575B1 (ko) * 2017-11-03 2020-06-24 영진약품 주식회사 멀꿀 잎 유래 추출물 또는 분획 정제물 또는 이로부터 분리된 신규 플라보노이드 화합물 및 카페인산 화합물을 유효성분으로 포함하는 항염, 또는 골조직 생성 또는 연골조직 생성 촉진용 약학적 조성물
KR102625435B1 (ko) 2023-03-17 2024-01-18 (주)제이비 바이오 천연발효효소복합액을 이용한 멀꿀 발효물과 이를 이용한 골밀도 개선, 뼈 성장촉진, 키 성장촉진 또는 관절건강증진용 조성물

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20150002398A (ko) * 2013-06-30 2015-01-07 재단법인 전남생물산업진흥원 멀꿀 잎 추출물을 유효성분으로 포함하는 골조직 생성 촉진용 약학 조성물
KR101652073B1 (ko) * 2015-10-12 2016-08-29 충남대학교산학협력단 멀꿀 추출물을 함유하는 전립선 질환의 예방 및 치료용 조성물

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR940011917B1 (ko) 1990-09-05 1994-12-27 태평양화학 주식회사 인삼잎 플라보노이드 성분의 추출방법
KR960004022B1 (ko) 1992-09-23 1996-03-25 1996년03월25일 은행잎에서 플라보노이드 화합물을 추출 정제하는 방법
KR100465778B1 (ko) 2002-05-30 2005-01-13 우리홍화인영농조합법인 항산화성 플라보노이드를 함유하고 있는 고품질의홍화잎차 추출물의 제조방법
KR100514916B1 (ko) 2003-02-10 2005-09-14 부경대학교 산학협력단 항산화활성을 갖는 연수 추출물 및 그로부터 분리된플라보노이드 화합물을 함유한 조성물
KR20040108265A (ko) 2003-06-17 2004-12-23 학교법인 한림대학교 플라보노이드를 포함하는 항-죽상경화증 조성물
KR100642151B1 (ko) 2004-01-14 2006-11-02 이호섭 돌나물 추출물로부터 분리한 플라보노이드 화합물들을포함하는 고혈압의 예방 및 치료용 약학조성물
KR100930927B1 (ko) 2008-03-11 2009-12-10 고려대학교 산학협력단 들깻잎으로부터 γ―ＧＣＳ의 활성을 증가시키는카페인산을 제조하는 방법, 그로부터 제조된 카페인산, 및상기 카페인산을 포함하는 식품 조성물 및 약학 조성물
KR20100043882A (ko) 2008-10-21 2010-04-29 김정환 헬리코박터 파이로리 균에 항균능을 갖는 뽕잎추출물, 이의제조방법 및 이를 이용한 건강기능식품

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20150002398A (ko) * 2013-06-30 2015-01-07 재단법인 전남생물산업진흥원 멀꿀 잎 추출물을 유효성분으로 포함하는 골조직 생성 촉진용 약학 조성물
KR101652073B1 (ko) * 2015-10-12 2016-08-29 충남대학교산학협력단 멀꿀 추출물을 함유하는 전립선 질환의 예방 및 치료용 조성물

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CHEON, Y.-H.: "Stauntonia hexaphylla (Lardizabalaceae) leaf methanol extract inhibits osteoclastogenesis and bone resorption activity via proteasome-mediated degradation of c-Fos protein and suppression of NFATcl expression", BMC COMPLEMENTARY AND ALTERNATIVE MEDICINE, 2015, pages 1 - 9 *
PARK, YUN-JUAN ET AL,: "Chemical Compound and Biological Activity of Stauntonia Hexaphylla", THE KOREAN JOURNAL OF PLANT RESOURCES, vol. 22, no. 5, 1 October 2009 (2009-10-01), pages 403 - 411, XP055516969 *
ZHAO, J.: "Cytotoxic Components in an Extract from the Leaves and Stems of Stauntonia hexaphylla", NATURAL PRODUCT SCIENCES, 2014, pages 130 - 134, XP053028801 *

Also Published As

Publication number Publication date
KR102038107B1 (ko) 2019-10-29
KR20180077997A (ko) 2018-07-09

Similar Documents

Publication Publication Date Title
EP2407160B1 (fr) Composition pharmaceutique pour la prévention et/ou le traitement de maladies osseuses, aliment fonctionnel ou aliment sain comprenant la composition et préparation pharmaceutique comprenant la composition en tant qu'ingrédient actif
KR102206591B1 (ko) 섬유화증 예방, 개선 또는 치료용 조성물
WO2018124695A1 (fr) Nouveau composé flavonoïde séparé de l'extrait de feuilles de stauntonia hexaphylla et composé pour favoriser l'anti-inflammation, formation osseuse ou formation de cartilage ayant la même fonction qu'un principe actif
KR102038108B1 (ko) 멀꿀 잎 추출물로부터 분리된 신규한 카페인산 화합물 및 이를 유효성분으로 하는 항염, 골조직 생성 또는 연골조직 생성 촉진용 조성물
KR101493461B1 (ko) 백미 추출물의 디옥시퍼굴라리닌을 유효성분으로 하는 결핵치료용 약학조성물
KR20110035127A (ko) 곰피와 감태 추출물 유래의 플로로탄닌을 유효성분으로 하는 염증억제용 조성물
KR101449575B1 (ko) 테뉴폴리사이드 ａ를 유효성분으로 포함하는 염증성 질환의 예방 또는 치료용 조성물
KR101168379B1 (ko) 가시오갈피 열매 추출물을 포함하는 통풍, 고혈압 및 암의 예방 및 치료용 조성물
KR20130005118A (ko) 사우치논을 유효성분으로 포함하는 근위축 억제 및 예방용 조성물
KR101418164B1 (ko) 자외선을 처리한 벼 추출물을 유효성분으로 포함하는 대장암 예방 또는 치료용 약학적 조성물
JP4472281B2 (ja) ラン科植物の含有成分とその用途
KR100833654B1 (ko) 삼백초 추출물 또는 이로부터 분리된 활성 화합물을포함하는 골다공증 예방 또는 치료용 조성물
KR20070019440A (ko) 후피향나무 잎으로부터 분리된 신규 화합물 및 이를 이용한항산화제
KR102350036B1 (ko) 양제근 추출물에서 유래한 화합물을 유효 성분으로 포함하는 혈액암 예방, 개선 및 치료용 조성물
KR101470613B1 (ko) 라티폴린을 유효성분으로 포함하는 염증성 질환의 예방 또는 치료용 조성물
KR102556835B1 (ko) 어수리 추출물을 유효성분으로 포함하는 면역증진 및 항비만용 조성물
WO2018062895A1 (fr) Composition comprenant de l'osmundacétone ou un sel pharmaceutiquement acceptable de cette dernière et destinée à la prévention ou au traitement de maladies osseuses
JP2008001673A (ja) ジオスシンを有効成分として含む糖吸収抑制用または胃粘膜保護用組成物およびジオスシン類似化合物
JP4024128B2 (ja) ウコギ科植物の花部の抽出物とその用途
KR20120054577A (ko) 곰피와 감태 추출물 유래의 플로로탄닌을 유효성분으로 하는 염증억제용 조성물
JP2024008233A (ja) フロログルシノール誘導体
JP2024082894A (ja) フロログルシノール誘導体
KR20150144370A (ko) 오디 유래 아릴벤조퓨란 및 플라보노이드 화합물을 포함하는 퇴행성 뇌질환의 예방, 개선 및 치료용 조성물
KR20240042861A (ko) 개다래 유래의 쿠마로일 트라이테르펜 유도체 및 이의 당뇨병 예방 또는 치료 용도
KR20240143369A (ko) 인가목 추출물을 포함하는 면역증진 및 항비만용 조성물

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17888915

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 17888915

Country of ref document: EP

Kind code of ref document: A1