WO2014007447A1 - Composition de prévention ou de traitement de maladies provoquées par l'angiogenèse, contenant un composé hydroxychalcone comme principe actif - Google Patents

Composition de prévention ou de traitement de maladies provoquées par l'angiogenèse, contenant un composé hydroxychalcone comme principe actif Download PDF

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WO2014007447A1
WO2014007447A1 PCT/KR2012/011867 KR2012011867W WO2014007447A1 WO 2014007447 A1 WO2014007447 A1 WO 2014007447A1 KR 2012011867 W KR2012011867 W KR 2012011867W WO 2014007447 A1 WO2014007447 A1 WO 2014007447A1
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hydroxyphenyl
propenone
disease
phenylpropenone
phenyl
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Korean (ko)
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이응석
김정애
강유라
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영남대학교 산학협력단
한국보건산업진흥원
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    • 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
    • A61K31/12—Ketones
    • 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
    • A61K31/12—Ketones
    • A61K31/122—Ketones having the oxygen directly attached to a ring, e.g. quinones, vitamin K1, anthralin
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/357—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having two or more oxygen atoms in the same ring, e.g. crown ethers, guanadrel
    • A61K31/36—Compounds containing methylenedioxyphenyl groups, e.g. sesamin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention relates to a composition for the prevention or treatment of diseases caused by angiogenesis, which contains a hydroxy baccon compound having angiogenic inhibitory activity as an active ingredient.
  • the present invention relates to a composition for the prevention or treatment of diseases caused by angiogenesis, which contains a hydroxy baccon compound having angiogenic inhibitory activity as an active ingredient.
  • Angiogenesis is a process in which new capillaries are formed from existing microvascular vessels.
  • angiogenesis occurs normally, embryonic development, tissue regeneration and wound healing, and periodic changes in the female genital system are called corpus luteum. Is a developmental stage, and in this case, it is strictly controlled (Folkman J et al., Int. Rev. Exp. Pathol. , 16, pp 207-248, 1976).
  • vascular endothelial cells grow very slowly and do not divide relatively well compared to other types of cells.
  • the angiogenesis process is generally caused by stimulation of angiogenesis factors, resulting in the formation of lumen due to the degradation of the basal membrane, proliferation, proliferation, and differentiation of vascular endothelial cells due to proteases. Consists of being generated.
  • angiogenesis there are diseases caused by angiogenesis that is not controlled autonomously and grows pathologically.
  • Diseases related to angiogenesis in pathological conditions include hemangioma, angiofibroma, angioplasty and cardiovascular diseases such as atherosclerosis, angiogenesis, and edema sclerosis.
  • Eye diseases caused by angiogenesis include corneal graft angiogenesis and angiogenesis. Glaucoma, diabetic retinopathy, corneal disease caused by neovascularization, spot degeneration, pterygium, retinal degeneration, posterior capsular fibrosis, granular conjunctivitis and the like.
  • Chronic inflammatory diseases such as arthritis, psoriasis, capillary dilatation, purulent granulomas, seborrheic dermatitis, dermatological diseases such as acne, Alzheimer's and obesity are also associated with angiogenesis, and cancer growth and metastasis necessarily depends on angiogenesis (D'Amato RJ et al., Ophthalmology, 102 (9), pp1261-1262, 1995; Arbiser JL, J. Am. Acad. Dermatol., 34 (3), pp486-497, 1996; O'Brien KD et al. Circulation, 93 (4), pp 672-682, 1996; Hanahan D et al., Cell, 86 , pp 353-364, 1996).
  • Angiogenesis plays an important role in the growth and metastasis of cancer cells.
  • Tumors are supplied with nutrients and oxygen for growth and proliferation through neovascularization, and new blood vessels that penetrate the tumors allow cancer cells to metastasize by giving the metastasizing cancer cells an opportunity to enter the blood circulation (Folkman and Tyler, Cancer Invasion and metastasis , Biologic mechanisms and Therapy (SB Day ed.) Raven press, New York, pp 94-103, 1977; Polverini PJ, Crit. Rev. Oral. Biol. Med. , 6 (3), pp 230-247, 1995).
  • the main cause of death of cancer patients is metastasis, and the current metastasis of cancer does not contribute to the survival of cancer patients.
  • diabetic retinopathy is a complication of diabetes, in which capillaries in the retina invade the vitreous body and eventually become blind.
  • Psoriasis which is characterized by red spots and skin, is also a chronic proliferative disease of the skin. It does not heal and involves pain and malformations. In normal cases, keratinocytes proliferate once a month, whereas psoriasis patients proliferate at least once a week. This rapid proliferation requires a lot of blood supply and angiogenesis is inevitably active (Folkman J, J. Invest. Dermatol., 59, pp 40-48, 1972).
  • Inflammatory bowel disease one of the other representative diseases of inflammatory diseases, is a term used to refer to diseases that cause inflammation of the intestinal ulcerative colitis, Crohn's disease, Behcet's disease, intestinal tuberculosis, amoeba enteritis, radiation enteritis, and drug-induced enteritis.
  • inflammatory bowel disease refers to ulcerative colitis and Crohn's disease, which are chronic idiopathic inflammatory diseases.
  • Ulcerative colitis is a disease in which mucous membranes or ulcers form continuously on the mucous membrane of the colon. Bloody stools, mucosal stools, diarrhea and abdominal pain occur, and in severe cases, systemic symptoms such as fever, weight loss, and anemia appear. .
  • Crohn's disease is a condition in which lesions such as ulcers are discontinuously occurring in any part of the digestive tract from the mouth to the anus. In addition to abdominal pain, diarrhea and bloody stools, in severe cases, fever, bleeding, weight loss, and general malaise , Anemia and other symptoms appear. Ulcerative colitis and Crohn's disease are both chronic refractory diseases that temporarily improve symptoms and then recur. The pathogenesis of these diseases is pointed out that environmental factors such as genetic factors, immune abnormalities, and meals are involved, but the cause is still unclear.
  • sulfasalazine which is frequently used as an aminosalicylic acid agent, has been reported to have side effects such as nausea, vomiting, anorexia, rash, headache, liver injury, white blood cell reduction, abnormal red blood cells, proteinuria, and diarrhea.
  • Prednisolone, an corticosteroid is used for oral administration, enema, suppository, intravenous injection, etc., but side effects such as gastric ulcer and femoral head necrosis due to long-term use are strong.
  • Infliximab a TNF-a monoclonal antibody
  • TNF Tumor Necrosis Factor
  • the inflammatory response is a mechanism for repairing and repairing the damaged area when an invasion that causes any organic change, such as physical action, chemicals, or bacterial infection, is applied to the living body or tissue.
  • Angiogenesis and vascular active substances such as bradykinin, prostagladin, and leukotriene are released to increase vascular permeability and cause inflammation, and excessive inflammatory reactions promote mucosal damage, and in some cases cancer Results in disease.
  • angiogenesis inhibitors can be applied as a therapeutic agent for such various angiogenesis-related diseases. Recently, studies to treat the above diseases by inhibiting angiogenesis have been actively conducted.
  • angiogenesis inhibitors generally require long-term administration to patients, and therefore, there is a need for development of drugs having low toxicity as angiogenesis inhibitors because they should be used as the most ideal therapeutic agent if they are less toxic and oral.
  • the present inventors confirmed that the hydroxy bacconium compounds of the present invention strongly inhibit angiogenesis according to treatment of angiogenesis inducers such as vascular endothelial growth factor in chicken choriocapillary model, thereby treating diseases caused by angiogenesis. And the present invention was completed by confirming that it can be usefully used as a preventive composition.
  • the disease caused by the angiogenesis is macular degeneration associated with aging, proliferative vitreoretinopathy, immature retinopathy, cone cornea, Sjogren's syndrome, corneal transplant rejection, abnormal wound fusion, bone disease, Proteinuria, abdominal aortic aneurysm, degenerative cartilage loss due to traumatic joint injuries, demyelination of the nervous system, cirrhosis, renal glomeruli, immature rupture of the embryonic membrane, periodontal membrane disease, cancer, inflammatory diseases, arthritis and bone metabolism Can be.
  • the cancer may be liver cancer, stomach cancer, colon cancer, lung cancer, breast cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer or myopic ophthalmic tumor.
  • the inflammatory disease may be gastritis, colitis, nephritis, hepatitis or inflammatory bowel disease.
  • the inflammatory bowel disease may be selected from the group consisting of Crohn's disease, intestinal lesions associated with Behcet's disease, ulcerative colitis, hemorrhagic rectal ulcer and ileal cystitis.
  • the bone metabolic disease is osteoporosis, Paget's bone disease, rickets disease, osteomalacia, nephrotic dystrophy in patients with renal failure, rheumatoid bone disease, degenerative bone disease, bone metastatic lesions, It may be a tumor, periodontal disease, inflammatory alveolar bone disease or an inflammatory bone resorption disease that is primarily produced in bone.
  • the compound is angiogenesis inhibitory effect by VEGF; Reduced inflammatory cytokine TNF- ⁇ (tumor necrosis factor- ⁇ ) and IL-17 production; And osteoclast differentiation inhibitory effect.
  • the compound may be included in a concentration of 0.1uM to 20uM in the composition.
  • the present invention is 1- (2-hydroxyphenyl) -3-phenylpropenone, 1- (3-hydroxyphenyl) -3-phenylpropenone, 1- (4-hydroxyphenyl) -3- Phenylpropenone, 3- (2-hydroxyphenyl) -1-phenylpropenone, 3- (3-hydroxyphenyl) -1-phenylpropenone, 3- (4-hydroxyphenyl) -1-phenylpropenone, 1, 3-bis (2-hydroxyphenyll) propenone, 1- (2-hydroxyphenyl) -3- (3-hydroxyphenyl) propenone, 1- (2-hydroxyphenyl) -3- (4 -Hydroxyphenyl) propenone, 1- (2-hydroxyphenyl) -1- (3-hydroxyphenyl) propenone, 1,3-bis (3-hydroxyphenyll) propenone, 1- (3 -Hydroxyphenyl) -3- (4-hydroxyphenyl) propenone, 3- (2-hydroxyphenyl) -1- (4-
  • the health functional food may be in the form of powder, granules, tablets, capsules, candy, beverages, gum, tea or vitamin complexes.
  • the present invention relates to a composition containing a hydroxy beacon compound that inhibits angiogenesis as an active ingredient, the compounds of the present invention is excellent in inhibiting angiogenesis in chorionic ureteric model, excellent anticancer activity, inflammatory response
  • the compounds of the present invention is excellent in inhibiting angiogenesis in chorionic ureteric model, excellent anticancer activity, inflammatory response
  • it also has the effect of inhibiting osteoclast differentiation, which is useful for the preparation of pharmaceutical compositions and health functional foods for the prevention and treatment of diseases caused by angiogenesis. have.
  • Figure 1 is a will showing the inhibitory effect of the compounds 10 to 15 according to the invention for the angiogenesis induced by VEGF (* P ⁇ 0.05 compared with the control group; comparison * P ⁇ 0.05 and VEGF- treated group),
  • Figure 2 shows the inhibitory effect of compounds 16-24 according to the invention on the angiogenesis induced by VEGF (compared with the control group * P ⁇ 0.05; compared * P ⁇ 0.05 and VEGF- treated group).
  • Figure 3 is a result showing the angiogenesis and tumor growth inhibitory effect of the compound according to the invention.
  • Figure 4 shows the compound inhibitory effect according to the invention on the inflammatory response induced by TNF- ⁇ .
  • Figure 5 is a result of measuring whether there is a change in body weight and treatment of the compound of the present invention in TNBS-induced inflammatory growth disease-induced animal model.
  • Figure 6 shows the results obtained by extracting the colon of the test animal after administration of TNBS for 5 days.
  • Figure 7 is the result of measuring the tissue weight after extracting the colon of the test animal after administration of TNBS for 5 days.
  • Figure 8 is the result of measuring the MPO activity in the intestinal mucosa to determine the degree of infiltration of inflammatory cells as an indicator for enteritis.
  • the present invention is characterized in that it provides a novel use of the hydroxy baccon compound as a new therapeutic agent that can effectively prevent and treat diseases caused by angiogenesis without side effects.
  • Diseases caused by angiogenesis as defined herein include macular degeneration associated with aging, proliferative vitreoretinopathy, immature retinopathy, keratoconus, Sjogren's syndrome, corneal transplant rejection, abnormal wound union, bone disease, proteinuria, abdominal aortic aneurysm , Degenerative cartilage loss due to traumatic joint injury, myelin deficiency diseases of the nervous system, cirrhosis, renal glomeruli, immature rupture of the embryonic membrane, periodontal membrane disease, cancer, inflammatory diseases, arthritis and bone metabolism diseases.
  • the cancer may be liver cancer, stomach cancer, colon cancer, lung cancer, breast cancer, rectal cancer, pancreatic cancer, brain cancer, skin cancer or myopic eye tumor, and the inflammatory disease may be gastritis, colitis, nephritis, hepatitis or inflammatory bowel disease.
  • the inflammatory bowel disease may include Crohn's disease, intestinal lesions associated with Behcet's disease, ulcerative colitis, hemorrhagic rectal ulcers and ileal cystitis, and the bone metabolic diseases include osteoporosis, Paget's bone disease, rickets, osteomalacia, kidney failure patients.
  • the compounds of the present invention can be prepared with pharmaceutically acceptable salts and solvates according to methods conventional in the art.
  • salts are acid addition salts formed with pharmaceutically acceptable free acids.
  • Acid addition salts are prepared by conventional methods, for example by dissolving a compound in an excess of aqueous acid solution and precipitating the salt using a water miscible organic solvent, such as methanol, ethanol, acetone or acetonitrile. Equivalent molar amounts of the compound and acid or alcohol (eg, glycol monomethyl ether) in water can be heated and the mixture can then be evaporated to dryness or the precipitated salts can be suction filtered.
  • a water miscible organic solvent such as methanol, ethanol, acetone or acetonitrile.
  • Equivalent molar amounts of the compound and acid or alcohol (eg, glycol monomethyl ether) in water can be heated and the mixture can then be evaporated to dryness or the precipitated salts can be suction filtered.
  • organic acids and inorganic acids may be used as the free acid, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, etc. may be used as the inorganic acid, and methanesulfonic acid, p -toluenesulfonic acid, acetic acid, trifluoroacetic acid, Citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid ( gluconic acid), galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanic acid, hydroiodic acid, and the like.
  • Bases can also be used to make pharmaceutically acceptable metal salts.
  • An alkali metal or alkaline earth metal salt is obtained by, for example, dissolving a compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and then evaporating and drying the filtrate.
  • the metal salt it is particularly suitable to prepare sodium, potassium or calcium salts, and the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).
  • Pharmaceutically acceptable salts of the compounds of the invention include salts of acidic or basic groups which may be present in the compound, unless otherwise indicated.
  • pharmaceutically acceptable salts include sodium, calcium and potassium salts of the hydroxy group
  • other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfate, hydrogen sulphate, phosphate, hydrogen phosphate, dihydrogen Phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p -toluenesulfonate (tosylate) salts, and methods or processes for preparing salts known in the art It can be prepared through.
  • Another object of the present invention is a method for preparing a compound of the present invention, can be prepared by a synthetic method known in the art, can be chemically synthesized by the method shown in the following schemes, but is not limited only to these examples no.
  • aryl methyl aldehyde benzaldehyde
  • EtOH EtOH
  • R2 ad
  • the compound of the present invention obtained by the above method is confirmed to strongly inhibit neovascularization following treatment of neovascularization inducer such as vascular endothelial growth factor in chicken choriocapillary model to treat diseases caused by angiogenesis. And it was confirmed that it can be usefully used as a preventive composition.
  • neovascularization inducer such as vascular endothelial growth factor in chicken choriocapillary model to treat diseases caused by angiogenesis.
  • composition of the present invention comprises 0.01 to 99% by weight of the compound relative to the total weight of the composition.
  • composition as described above is not necessarily limited thereto, and may vary according to the condition of the patient and the type and extent of the disease.
  • compositions comprising a compound of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.
  • compositions comprising the compounds according to the invention are each formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories, and sterile injectable solutions according to conventional methods.
  • the carriers, excipients and diluents that may be included in the formulation may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium Silicates, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate and mineral oil.
  • diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used.
  • Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one or more excipients in the compound, at least cotton, starch, calcium carbonate, sucrose. Or lactose, gelatin, or the like is mixed. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Oral liquid preparations include suspending agents, liquid solutions, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. .
  • Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories.
  • non-aqueous solvent and suspending agent propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used.
  • base of the suppository witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
  • Preferred dosages of the compounds of the present invention depend on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, but may be appropriately selected by those skilled in the art.
  • the compound is preferably administered at 0.01 mg / kg to 10 g / kg per day, preferably at 1 mg / kg to 1 g / kg. Administration may be administered once a day or may be divided several times. Therefore, the above dosage does not limit the scope of the present invention in any aspect.
  • composition of the present invention can be administered to mammals such as rats, mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example by oral and rectal or intravenous methods.
  • the present invention also provides 1- (2-hydroxyphenyl) -3-phenylpropenone, 1- (3-hydroxyphenyl) -3-phenylpropenone, 1- (4-hydroxyphenyl) -3-phenylpropenone, 3- (2-hydroxyphenyl) -1-phenylpropenone, 3- (3-hydroxyphenyl) -1-phenylpropenone, 3- (4-hydroxyphenyl) -1-phenylpropenone, 1,3-bis (2-hydroxyphenyll) propenone, 1- (2-hydroxyphenyl) -3- (3-hydroxyphenyl) propenone, 1- (2-hydroxyphenyl) -3- (4-hydroxyphenyl) propenone, 1- (2-hydroxyphenyl) -1- (3-hydroxyphenyl) propenone, 1,3-bis (3-hydroxyphenyll) propenone, 1- (3-hydroxyphenyl) -3- (4-hydroxyphenyl) propenone, 3- (2-hydroxyphenyl) -1- (4-hydroxyphenyl) propenone, 3-
  • Health functional foods containing the compound of the present invention can be used in a variety of drugs, foods and beverages for the prevention and improvement of diseases caused by angiogenesis.
  • examples of the food to which the compound of the present invention may be added include various foods, beverages, gums, teas, vitamin complexes, health supplements, and the like, and may be used in the form of powders, granules, tablets, capsules, or beverages. have.
  • the present invention also provides 1- (2-hydroxyphenyl) -3-phenylpropenone and 1- (3-hydroxyphenyl) -3-phenylpropenone having the effect of preventing and improving diseases caused by angiogenesis.
  • the food form to which the compound of the present invention can be added includes various foods, beverages, gums, teas, vitamin complexes, or health supplements of candy.
  • the compounds of the present invention may be added to foods or beverages for the purpose of preventing and ameliorating diseases caused by angiogenesis.
  • the amount of the compound in the food or beverage is generally added to the health food composition of the present invention to 0.01 to 15% by weight of the total food weight, the health beverage composition is 0.02 to 10 g, preferably based on 100 ml Can be added in a ratio of 0.3 to 1 g.
  • the health beverage composition of the present invention contains a mixture of the above compounds as essential ingredients in the ratios indicated, and there are no particular limitations on the liquid components, and it can contain various flavors or natural carbohydrates as additional ingredients, like ordinary drinks.
  • natural carbohydrates are conventional monosaccharides such as disaccharides such as glucose and fructose, such as maltose, sucrose and the like, and polysaccharides such as dextrin, cyclodextrin and the like.
  • Sugars and sugar alcohols such as xylitol, sorbitol, and erythritol.
  • natural flavoring agents such as, tauumatin, stevia compounds (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used.
  • the proportion of said natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.
  • the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like.
  • the compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.
  • HPLC analysis was performed using two instruments (Shimadzu LC-10AT pumps gradient-controlled HPLC system, Shimadzu system controller (SCL-10A VP) and photo diode array detector (SPD-M10A VP), using Shimadzu Class VP program).
  • Shimadzu system controller SCL-10A VP
  • SPD-M10A VP photo diode array detector
  • volumetric sample (10 ⁇ L) was subjected to constant conditions (bevel dilution: 50% to 100% of B in A and 100% to 50% of B in A for 10 minutes Flow rate: 1.0 mL / min, t 254 nm UV detection Mobile phase A was injected into a column of X-Terra 5 ⁇ Mreverse-phase C 18 column (4.6 ⁇ 250 mm) in 20 mM ammonium formate (AF) treated secondary distilled water and B is 90% ACN solvent in 20 mM AF treated water. Purity is expressed in percent (%).
  • acetophenone acetophenone, 1.16 mL, 10.00 mmol
  • 3-hydroxybenzaldehyde hydroxybenzaldehyde, 1.22 g, 10.00 mmol
  • 3-hydroxyphenyl -1-phenylpropenone (2.11 g, 94.1%, 9.4 mmol) was obtained.
  • the fertilized egg was purchased from a white chicken farm (Cheongsong, Korea), and cortisone acetate was purchased from Aldrich Chemical Co., St. Louis, MO, USA.
  • cortisone acetate was purchased from Aldrich Chemical Co., St. Louis, MO, USA.
  • vascular endothelial growth factor was purchased from R & D systems, Minneapolis, MN, USA and whatman filter disc was purchased from Whatman, UK.
  • chorioallantoic membrane (CAM) analysis was performed (Nguyen M et al., Microvascular Res. , 47, pp31-40, 1994). .
  • hypodermic needle hypodermic needle, Green Cross Medical Industry, Korea
  • the villus urea separates from the shell of the fertilized egg, which is then cut with a grinding wheel (Multipro 395JA, Dremel, Mexico). A cm 2 sized window was made.
  • VEGF vascular endothelial growth factor
  • the CAM part on the filter disc was removed and washed with phosphate buffer solution, followed by a stereo microscope (Stemi SV6 stereomicroscope, Carl Zeiss, Germany) and Image-Pro Plus software (Media Cybernetics; Silver Spring, MD, USA) was used to measure the number of vascular branches and analyze data.
  • a stereo microscope Stemi SV6 stereomicroscope, Carl Zeiss, Germany
  • Image-Pro Plus software Media Cybernetics; Silver Spring, MD, USA
  • the compounds according to the present invention strongly inhibit VEGF-induced neovascularization in vivo (see Table 1), and the blood vessel branch points were added to the PBS-treated group. In comparison, it was significantly increased by VEGF.
  • Treatment of the compounds of the present invention against CAM significantly inhibited VEGF-induced neovascularization in terms of number and small vessel length after incubation (see FIGS. 1A, 1B, and 2A, 2B).
  • Compound 15 showed the strongest anti-angiogenic activity in vivo at 1 / CAM (66%).
  • Compound 19 also showed the strongest inhibitory activity (89%) among the compounds, and similarly compounds 21 and 22 also showed the strong inhibitory activity (80% and 87%, respectively).
  • the present inventors confirmed that 15 compounds of the present invention have high angiogenesis inhibitory effects in vivo through Experimental Example 1, and furthermore, the following experiments were performed to confirm whether these compounds actually have anticancer activity.
  • the HT-29 human-derived colorectal cancer cells were mixed 1: 1 with matrigel instead of discs, and then, among the compounds synthesized in Examples of the present invention.
  • Hydroxychalcone compound NO. 19, 21 and 22 were treated with colorectal cancer cells, respectively, wherein the cells were seeded at a density of 1.5 ⁇ 10 6 cells / CAM.
  • the tumor-formed CAM was removed and washed with phosphate buffer solution, followed by stereomicroscopy (Stemi SV6 stereomicroscope, Carl Zeiss, Germany) and Image-Pro Plus software (Media Cybernetics; Silver Spring, MD, USA).
  • stereomicroscopy Stereomicroscopy
  • Carl Zeiss Carl Zeiss, Germany
  • Image-Pro Plus software Media Cybernetics; Silver Spring, MD, USA.
  • angiogenesis which plays a role in tumor formation, was inhibited by the treatment of the compounds of the present invention NO.19, 21, and 22, and tumor growth was also inhibited.
  • the present inventors found that the 15 compounds according to the present invention not only effectively inhibit tumor-induced angiogenesis, but also inhibit tumor growth, and thus can be used as a new anticancer agent.
  • the 15 compounds according to the present invention not only effectively inhibit tumor-induced angiogenesis, but also inhibit tumor growth, and thus can be used as a new anticancer agent.
  • HT-29 human colon epithelial cells and U937 human monocytes were used as cellular models of inflammatory bowel disease.
  • HT-29 cells (American Type Culture Collections, Rockville, Mass., USA) were cultured in RPMI 1640 containing 10% fetal bovine serum (FBS), 1% penicillin / streptomycin and 2 mmol / L glutamine, 95 The cell line was incubated at 37 ° C. under an environment of% air and 5% CO 2. The following experiments used cells of 36 passages or less, and cells were subcultured weekly using Dulbecco's phosphate buffered saline (D-PBS) containing 0.25% trypsin and 1% EDTA. The culture medium was replaced every two days and allowed to grow confluent on the culture plate, then subcultured in a 1: 5 ratio.
  • D-PBS Dulbecco's phosphate buffered saline
  • Each cell was No. 1 in the compound prepared in Example 1 1 hour before stimulation of TNF- ⁇ . 19 compounds were used and the stock solution of each compound was dissolved in dimethylsulfoxide (DMSO). At this time, the final maximum concentration of DMSO used in the test medium was 0.1% or less.
  • DMSO dimethylsulfoxide
  • cells treated with the control group and TNF- ⁇ alone were pretreated with a test medium containing 0.1% DMSO, and 20 mM 5-aminosalicylic acid (5-ASA) was used as a positive control. Disease) is known to have the effect of inhibiting the activated inflammatory sequence.
  • the compound according to the present invention has the effect of inhibiting the adhesion of TNF- ⁇ -induced intestinal epithelial cells, which ultimately has the effect of preventing and inhibiting enteritis, ie, inflammatory diseases. I could see that.
  • the present inventors performed the following experiments on animal models of inflammatory bowel disease to determine whether the hydroxy chancon compound of the present invention has a therapeutic effect on inflammatory bowel disease.
  • the test animal was purchased from Samtaco Bio Korea for 7 days old Sprague Dawley species, stabilized with a general solid feed for 2 days, and used for experiments. It was supplied freely, the temperature of the kennel was maintained at 25 ⁇ 1 °C, relative humidity 50 ⁇ 10%. Lighting control was controlled by a 12-hour light-dark cycle by an automatic light controller.
  • the experimental group consisted of 6 animals in each group, and the average weight was 180 ⁇ 10 g by the randomized block design (5 groups) (control group, TNBS alone group, TNBS + 5-ASA 100 mg / kg administration group, TNBS + hydrolock).
  • Siconcone Compound No. 19 was administered in 1 mg / kg administration group, TNBS + hydroxychalcone Compound No.19 administered in the 10 mg / kg administration group, TNBS + Chalcone 10 mg / kg administration group, and TNBS + Chalcone 50 mg / kg administration group).
  • TNBS trobencene sulfonic acid
  • the drug was intraperitoneally administered with hydroxychalcone compound No.19 at 1 mg / kg or 10 mg / kg once daily at a constant time for 5 days after TNBS treatment after 24 hours of fasting.
  • the comparative test material was 5-ASA and chalcone compounds, active metabolites of sulfasalazine, the best known agent for treating inflammatory bowel disease.
  • test animals were sacrificed 7 days after TNBS administration and the severity of visual ulcer and inflammatory bowel disease (colitis) was assessed by two investigators who did not participate in the experiment.
  • the large intestine of the test animal was removed, and tissues between 5 and 6 cm from the anus were cut into 1 cm lengths and used for measuring the intestinal weight and MPO activity of the tissues and performing a biopsy.
  • the present inventors examined the naked eye after 5 days of drug administration, and as a result, the large intestine of TNBS-treated test animals was observed edema and hyperemia compared to the control group, and edema and congestion and adhesion of appendicitis were observed. appear.
  • the group treated with hydroxychalcone compound No. 19 of the present invention was observed to be significantly reduced edema and hyperemia compared to the TNBS alone group, in particular hydroxychalcone compound 19 treated with 10 mg / kg
  • One group was observed to be similar in shape to the normal group without any treatment. Therefore, these results show that the present inventors can effectively inhibit the edema and hyperemia of the bowel caused by inflammatory bowel disease of the present invention. This effect is known as a therapeutic agent for inflammatory bowel disease. It was found that the therapeutic effect was better than that of ASA (see FIG. 6).
  • the weight of the intestinal edema was significantly increased in the TNBS-treated group compared to the vehicle-treated control group.
  • the intestinal weight was significantly decreased compared to the TNBS treatment group, and the group treated with hydroxychalcone compound 19 showed less relief than the control group.
  • the group treated with hydroxychalcone compound 19 showed less relief than the control group.
  • the present inventors measured the MPO activity in the intestinal mucosa to determine the degree of invasion of inflammatory cells as an indicator for enteritis.
  • MPO Myeloperoxidase
  • the inventors used an MPO assay kit to measure MPO activity.
  • the colon tissues were washed with cold PBS and weighed to determine the lysis buffer (pH7.4, 200 mM NaCl, 5 mM EDTA, 10 per 10 mg of tissue weight). 200 ⁇ l of mM Tris, 10% glycerol) was added and homogenized for 30 seconds using a tissue homogenizer (Bio homogenizer M133, BIOSPEC PRODUCTS Inc. USA).
  • the homogenized sample was centrifuged twice at 1500 x g for 15 minutes to obtain a supernatant, and then the supernatant was measured for MPO activity using an MPO ELISA kit (HK210, Hycult Biotechnology, Netherlands).
  • the MPO activity refers to a value at which 1 ⁇ M of hydrogen peroxide is reduced in water at 25 ° C. for 1 minute, and the colorectal tissue homogenate. Calculated by the amount of MPO contained in 1 ml.
  • MPO activity was significantly higher in the TNBS-treated group than in the control group, and MPO activity was significantly decreased in the 5-ASA 100 mg / kg administration group.
  • MPO activity was lower than that in the TNBS alone group, and especially 1 mg / kg inhibited the MPO activity of the tissues better than 5-ASA 100 mg / kg. It could be seen (see Fig. 8).
  • the above ingredients are mixed and filled in an airtight cloth to prepare a powder.
  • tablets are prepared by tableting according to a conventional method for preparing tablets.
  • the above ingredients are mixed and filled into gelatin capsules to prepare capsules.
  • the amount of the above ingredient is prepared per ampoule (2 ml).
  • Nicotinic Acid Amide ... 1.7 mg
  • composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method.
  • the granules may be prepared and used for preparing a health food composition according to a conventional method.
  • composition ratio is a composition that is relatively suitable for the preferred beverage in a preferred embodiment
  • compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

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Abstract

La présente invention concerne une composition contenant un composé hydroxychalcone, comme principe actif, et destinée à inhiber l'angiogenèse. Les composés selon la présente invention ont des effets remarquables d'inhibition de l'angiogenèse dans le modèle de membrane chorioallantoïque, et peuvent ainsi être utilisés comme composition pharmaceutique et comme complément alimentaire destinés à prévenir et à traiter des maladies provoquées par l'angiogenèse.
PCT/KR2012/011867 2012-07-03 2012-12-31 Composition de prévention ou de traitement de maladies provoquées par l'angiogenèse, contenant un composé hydroxychalcone comme principe actif WO2014007447A1 (fr)

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CN114380680A (zh) * 2022-01-20 2022-04-22 广州市朝利良生物科技有限公司 一种黄酮类化合物及其应用
CN115124409A (zh) * 2022-05-30 2022-09-30 山东科源制药股份有限公司 一种盐酸普罗帕酮中间体的制备方法

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KR101988757B1 (ko) * 2017-12-19 2019-06-12 가톨릭대학교 산학협력단 1,2-디하이드록시-3-메틸안트라퀴논을 유효성분으로 하는 간암 예방 또는 치료용 조성물

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CN114380680A (zh) * 2022-01-20 2022-04-22 广州市朝利良生物科技有限公司 一种黄酮类化合物及其应用
CN115124409A (zh) * 2022-05-30 2022-09-30 山东科源制药股份有限公司 一种盐酸普罗帕酮中间体的制备方法

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