KR101804109B1 - Chrysin having improved antinflammation ability by irradiation, method for producing the same and method - Google Patents
Chrysin having improved antinflammation ability by irradiation, method for producing the same and method Download PDFInfo
- Publication number
- KR101804109B1 KR101804109B1 KR1020150117307A KR20150117307A KR101804109B1 KR 101804109 B1 KR101804109 B1 KR 101804109B1 KR 1020150117307 A KR1020150117307 A KR 1020150117307A KR 20150117307 A KR20150117307 A KR 20150117307A KR 101804109 B1 KR101804109 B1 KR 101804109B1
- Authority
- KR
- South Korea
- Prior art keywords
- chrysin
- inflammatory
- disease
- irradiation
- present
- Prior art date
Links
- RTIXKCRFFJGDFG-UHFFFAOYSA-N chrysin Chemical compound C=1C(O)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC=CC=C1 RTIXKCRFFJGDFG-UHFFFAOYSA-N 0.000 title claims abstract description 141
- NYCXYKOXLNBYID-UHFFFAOYSA-N 5,7-Dihydroxychromone Natural products O1C=CC(=O)C=2C1=CC(O)=CC=2O NYCXYKOXLNBYID-UHFFFAOYSA-N 0.000 title claims abstract description 64
- 229940043370 chrysin Drugs 0.000 title claims abstract description 64
- 235000015838 chrysin Nutrition 0.000 title claims abstract description 64
- 238000000034 method Methods 0.000 title claims abstract description 24
- 238000004519 manufacturing process Methods 0.000 title abstract description 19
- -1 chrysin derivative compound Chemical class 0.000 claims abstract description 15
- 230000005855 radiation Effects 0.000 claims description 21
- 208000027866 inflammatory disease Diseases 0.000 claims description 15
- 230000002757 inflammatory effect Effects 0.000 claims description 11
- 235000013376 functional food Nutrition 0.000 claims description 10
- 239000004480 active ingredient Substances 0.000 claims description 7
- 230000001678 irradiating effect Effects 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 208000029147 Collagen-vascular disease Diseases 0.000 claims description 6
- 201000004624 Dermatitis Diseases 0.000 claims description 6
- 208000004232 Enteritis Diseases 0.000 claims description 6
- 208000007882 Gastritis Diseases 0.000 claims description 6
- 206010018364 Glomerulonephritis Diseases 0.000 claims description 6
- 206010068319 Oropharyngeal pain Diseases 0.000 claims description 6
- 206010033645 Pancreatitis Diseases 0.000 claims description 6
- 201000007100 Pharyngitis Diseases 0.000 claims description 6
- 206010035664 Pneumonia Diseases 0.000 claims description 6
- 206010038910 Retinitis Diseases 0.000 claims description 6
- 206010040047 Sepsis Diseases 0.000 claims description 6
- 206010003246 arthritis Diseases 0.000 claims description 6
- 206010006451 bronchitis Diseases 0.000 claims description 6
- 230000036541 health Effects 0.000 claims description 6
- 208000006454 hepatitis Diseases 0.000 claims description 6
- 231100000283 hepatitis Toxicity 0.000 claims description 6
- 201000008383 nephritis Diseases 0.000 claims description 6
- 201000000306 sarcoidosis Diseases 0.000 claims description 6
- 206010044008 tonsillitis Diseases 0.000 claims description 6
- 208000022559 Inflammatory bowel disease Diseases 0.000 claims description 5
- 238000000354 decomposition reaction Methods 0.000 claims description 4
- 238000011282 treatment Methods 0.000 claims description 4
- 239000008194 pharmaceutical composition Substances 0.000 claims description 3
- 206010057190 Respiratory tract infections Diseases 0.000 claims 3
- 235000013402 health food Nutrition 0.000 claims 1
- 238000011321 prophylaxis Methods 0.000 claims 1
- 230000003110 anti-inflammatory effect Effects 0.000 abstract description 30
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 abstract description 27
- 230000009466 transformation Effects 0.000 abstract description 5
- 230000004048 modification Effects 0.000 abstract description 2
- 238000012986 modification Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 17
- WDECIBYCCFPHNR-UHFFFAOYSA-N chrysene Chemical compound C1=CC=CC2=CC=C3C4=CC=CC=C4C=CC3=C21 WDECIBYCCFPHNR-UHFFFAOYSA-N 0.000 description 17
- 235000013305 food Nutrition 0.000 description 14
- 230000000694 effects Effects 0.000 description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 10
- 230000005251 gamma ray Effects 0.000 description 10
- 230000001965 increasing effect Effects 0.000 description 9
- 229930014626 natural product Natural products 0.000 description 8
- 102000004889 Interleukin-6 Human genes 0.000 description 7
- 108090001005 Interleukin-6 Proteins 0.000 description 7
- 235000013361 beverage Nutrition 0.000 description 7
- 230000008859 change Effects 0.000 description 7
- 238000011156 evaluation Methods 0.000 description 7
- 229940100601 interleukin-6 Drugs 0.000 description 7
- 230000001766 physiological effect Effects 0.000 description 7
- 102000004127 Cytokines Human genes 0.000 description 6
- 108090000695 Cytokines Proteins 0.000 description 6
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 6
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 6
- 239000002158 endotoxin Substances 0.000 description 6
- 238000004128 high performance liquid chromatography Methods 0.000 description 6
- 229920006008 lipopolysaccharide Polymers 0.000 description 6
- 206010061218 Inflammation Diseases 0.000 description 5
- 241001529936 Murinae Species 0.000 description 5
- 231100000987 absorbed dose Toxicity 0.000 description 5
- 231100000135 cytotoxicity Toxicity 0.000 description 5
- 230000003013 cytotoxicity Effects 0.000 description 5
- 238000011161 development Methods 0.000 description 5
- 230000018109 developmental process Effects 0.000 description 5
- XEYBRNLFEZDVAW-ARSRFYASSA-N dinoprostone Chemical compound CCCCC[C@H](O)\C=C\[C@H]1[C@H](O)CC(=O)[C@@H]1C\C=C/CCCC(O)=O XEYBRNLFEZDVAW-ARSRFYASSA-N 0.000 description 5
- 229960002986 dinoprostone Drugs 0.000 description 5
- 230000004054 inflammatory process Effects 0.000 description 5
- 230000002401 inhibitory effect Effects 0.000 description 5
- 210000002540 macrophage Anatomy 0.000 description 5
- XEYBRNLFEZDVAW-UHFFFAOYSA-N prostaglandin E2 Natural products CCCCCC(O)C=CC1C(O)CC(=O)C1CC=CCCCC(O)=O XEYBRNLFEZDVAW-UHFFFAOYSA-N 0.000 description 5
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 4
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 239000000796 flavoring agent Substances 0.000 description 4
- 235000019634 flavors Nutrition 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 239000003963 antioxidant agent Substances 0.000 description 3
- 230000003078 antioxidant effect Effects 0.000 description 3
- 235000006708 antioxidants Nutrition 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- PUPZLCDOIYMWBV-UHFFFAOYSA-N (+/-)-1,3-Butanediol Chemical compound CC(O)CCO PUPZLCDOIYMWBV-UHFFFAOYSA-N 0.000 description 2
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 2
- 108010037462 Cyclooxygenase 2 Proteins 0.000 description 2
- 238000008157 ELISA kit Methods 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 102100038280 Prostaglandin G/H synthase 2 Human genes 0.000 description 2
- 102100040247 Tumor necrosis factor Human genes 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 235000013351 cheese Nutrition 0.000 description 2
- 235000019219 chocolate Nutrition 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000010894 electron beam technology Methods 0.000 description 2
- 235000013373 food additive Nutrition 0.000 description 2
- 239000002778 food additive Substances 0.000 description 2
- 235000019253 formic acid Nutrition 0.000 description 2
- 235000020510 functional beverage Nutrition 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 235000013622 meat product Nutrition 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 229920001451 polypropylene glycol Polymers 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 241000251468 Actinopterygii Species 0.000 description 1
- KYNSBQPICQTCGU-UHFFFAOYSA-N Benzopyrane Chemical compound C1=CC=C2C=CCOC2=C1 KYNSBQPICQTCGU-UHFFFAOYSA-N 0.000 description 1
- 241001474374 Blennius Species 0.000 description 1
- 229940046168 CpG oligodeoxynucleotide Drugs 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 239000004278 EU approved seasoning Substances 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- 231100000002 MTT assay Toxicity 0.000 description 1
- 238000000134 MTT assay Methods 0.000 description 1
- 102000008299 Nitric Oxide Synthase Human genes 0.000 description 1
- 108010021487 Nitric Oxide Synthase Proteins 0.000 description 1
- 108010016731 PPAR gamma Proteins 0.000 description 1
- 229920002230 Pectic acid Polymers 0.000 description 1
- 102100038825 Peroxisome proliferator-activated receptor gamma Human genes 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 240000000697 Pinguicula vulgaris Species 0.000 description 1
- 244000294611 Punica granatum Species 0.000 description 1
- 235000014360 Punica granatum Nutrition 0.000 description 1
- 241000269821 Scombridae Species 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 239000000556 agonist Substances 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000003276 anti-hypertensive effect Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 235000008452 baby food Nutrition 0.000 description 1
- 235000013527 bean curd Nutrition 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 235000014171 carbonated beverage Nutrition 0.000 description 1
- 231100000357 carcinogen Toxicity 0.000 description 1
- 239000003183 carcinogenic agent Substances 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 239000010779 crude oil Substances 0.000 description 1
- 235000015140 cultured milk Nutrition 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 235000019985 fermented beverage Nutrition 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 235000019688 fish Nutrition 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 235000011194 food seasoning agent Nutrition 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 235000015243 ice cream Nutrition 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 208000026278 immune system disease Diseases 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000017306 interleukin-6 production Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 235000021109 kimchi Nutrition 0.000 description 1
- 235000020640 mackerel Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 231100001083 no cytotoxicity Toxicity 0.000 description 1
- 235000012149 noodles Nutrition 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 239000003002 pH adjusting agent Substances 0.000 description 1
- LCLHHZYHLXDRQG-ZNKJPWOQSA-N pectic acid Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)O[C@H](C(O)=O)[C@@H]1OC1[C@H](O)[C@@H](O)[C@@H](OC2[C@@H]([C@@H](O)[C@@H](O)[C@H](O2)C(O)=O)O)[C@@H](C(O)=O)O1 LCLHHZYHLXDRQG-ZNKJPWOQSA-N 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 229940127557 pharmaceutical product Drugs 0.000 description 1
- 230000003128 phytoestrogenic effect Effects 0.000 description 1
- 235000021110 pickles Nutrition 0.000 description 1
- 239000010318 polygalacturonic acid Substances 0.000 description 1
- 235000012015 potatoes Nutrition 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000003608 radiolysis reaction Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000033764 rhythmic process Effects 0.000 description 1
- 235000015067 sauces Nutrition 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 235000011888 snacks Nutrition 0.000 description 1
- 235000013555 soy sauce Nutrition 0.000 description 1
- 235000015096 spirit Nutrition 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000012916 structural analysis Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002277 temperature effect Effects 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 230000006433 tumor necrosis factor production Effects 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
Images
Classifications
-
- 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
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
- A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
-
- 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
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Mycology (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Nutrition Science (AREA)
- Engineering & Computer Science (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The present invention relates to a method for preparing chrysin having enhanced anti-inflammatory activity, and more particularly, to a method for producing chrysin by dissolving chrysin in methanol followed by transformation by irradiation with gamma rays to enhance anti-inflammatory activity. When producing chrysin by the production method according to the present invention, it is possible to produce a chrysin derivative compound that has undergone structural modification by irradiation, and the chrysin prepared by the above method has a superior anti-inflammatory activity Is effective.
Description
The present invention relates to a method for preparing chrysin having enhanced anti-inflammatory activity, and more particularly, to a method for producing chrysin by dissolving chrysin in methanol followed by transformation by irradiation with gamma rays to enhance anti-inflammatory activity.
Recently, as the quality of life has increased, interest in health has increased and the number of chronic complex diseases has been increasing due to aging population and western type change of lifestyle. The development of health functional foods / medicines with increased functionality based on long-used clinical experience has increased the importance of natural products due to changes in awareness for prevention and treatment. Research and development for the development of novel natural products / medicines that can prevent and treat various diseases while minimizing side effects while having excellent efficacy not only in domestic but also in the world are proceeding. Chrysin, one of the phytoestrogenic substances, is widely distributed in plants such as pomegranate, plum, bean, and seaweed, and is widely used as an antioxidant, antihypertensive, antioxidant, anti-cancer, anti- Various biological activities have been reported. Various methods such as synthesis of natural materials and enzymatic methods for enhancing the inherent physiological activity of natural products and developing materials have been studied. In the development of existing natural products, there is a high possibility that contaminants such as other impurities are contained because the specific ingredients are concentrated and heated. In recent years, there have been cases of safety controversy including detection of carcinogens such as benzopyran and formaldehyde. The use of radiation technology is required as a measure for the side effects of the development process.
In order to solve such a problem, the present invention relates to the development of a natural biomaterial-use / pharmaceutical product which enhances the intrinsic physiological activity (anti-inflammatory effect) of natural products possessed by chrysin through structural transformation of natural biological materials, The aim of this study was to prepare radiation - induced chrysene.
Accordingly, an object of the present invention is to provide a method for preparing a radiation-induced chrysene for producing chrysin having excellent anti-inflammatory activity and a structural transformed chrysin prepared by the method.
In order to achieve the above object, there is provided a method for producing a chrysin derivative having an anti-inflammatory activity, which comprises irradiating a chrysene compound with radiation.
In one embodiment of the present invention, the irradiation with radiation may be a step of dissolving the chrysene compound in alcohol and then irradiating it with radiation.
In one embodiment of the present invention, the alcohol may be selected from the group consisting of methanol, ethanol, n-propanol, isopropanol, ethylene glycol, polypropylene glycol, and butylene glycol.
In one embodiment of the present invention, the radiation may be gamma rays, X-rays, or radiation of electron beams.
In one embodiment of the present invention, the radiation is gamma rays, which may be irradiated at 15 to 50 kGy.
The present invention also provides a pharmaceutical composition for preventing or treating inflammatory diseases, which comprises the chrysin derivative compound prepared by the method according to the present invention as an active ingredient.
In one embodiment of the present invention the inflammatory disease is selected from inflammatory bowel disease, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, sore throat, bronchitis, pneumonia, pancreatitis , Sepsis, and nephritis.
The present invention also provides a health functional food for preventing or ameliorating an inflammatory disease which comprises the chrysin derivative compound prepared by the method according to the present invention as an active ingredient.
In one embodiment of the present invention the inflammatory disease is selected from inflammatory bowel disease, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, sore throat, bronchitis, pneumonia, pancreatitis , Sepsis, and nephritis.
When producing chrysin by the production method according to the present invention, it is possible to produce a chrysin derivative compound that has undergone structural modification by irradiation, and the chrysin prepared by the above method has a superior anti-inflammatory activity Is effective.
BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic diagram of a process for producing the chrysin derivative of the present invention. FIG.
FIG. 2 is a graph showing the results of HPLC analysis after irradiation of gamma-ray to chrysin dissolved in methanol.
Figure 3 shows the cytotoxicity evaluation results of the gamma-irradiated chrysin derivative of the present invention.
4 is a graph showing the results of measuring the inhibitory effect on the inflammatory factors NO and PEG2 of the gamma-irradiated chrysin derivative of the present invention.
(a) Evaluation results of NO production inhibition of gamma-irradiated chrysin derivatives.
(b) the inhibitory effect of gamma-irradiated chrysin derivatives on PEG2 production.
FIG. 5 is a graph showing the results of measuring the inhibitory effect of inflammatory cytokines on the gamma-irradiation chromin derivatives of the present invention.
(a) Evaluation of inhibition of IL-6 production by gamma-irradiation chrysin derivatives.
(b) the inhibition of TNF-α production by gamma-irradiation chrysin derivatives.
The present invention is characterized by providing a chrysin derivative in which the molecular structure is changed due to irradiation and the anti-inflammatory activity is enhanced, and a method for producing the same.
Chrysin (5,7-dihydroxyflavone) is a natural flavonoid that exhibits a wide range of physiological activities such as antioxidant and anti-inflammatory action. Since the anti-inflammatory mechanism of chrysin acts as an agonist of the PPAR-γ receptor, it inhibits the functions of cyclooxygenase-2 (COX-2) and in vivo nitric oxide synthase (iNOS), which are important inflammatory precursor enzymes .
The absorbed dose of radiation used in the present invention is 5 to 150 kGy, preferably 10 to 100 kGy, more preferably 15 to 50 kGy. When the absorbed dose is less than 5 kGy, The target can not be achieved. On the other hand, when the dose is more than 150 kGy, problems such as decomposition of the material by high dose of radiation may occur.
In order to evaluate the cytotoxicity of chrysin transformed by the irradiation of the present invention, 0, 3.125, 6.25, 12.5, and 25 μM of Murine macrophage RAW264.7 cells were treated to measure cell viability. As a result, it was confirmed that there was no cytotoxicity, and thus it was confirmed that the radiation-treated chrysin derivative prepared by the method of the present invention was a stable substance in the human body (see FIG. 3).
Meanwhile, in order to confirm the effect of increasing the anti-inflammatory activity compared to the non-irradiated chrysin of the chrysin having the molecular structure converted by the irradiation according to the present invention, the inhibitory activity of the anti-inflammatory activity factors NO and PEG2 was confirmed, It was confirmed that the irradiated chrysene significantly inhibited the production of NO and PEG2 as compared with non-irradiated chrysin (see Fig. 4).
Further, in order to confirm the effect of increasing the anti-inflammatory activity compared with the non-irradiated chrysin of the chrysin having the molecular structure converted by the irradiation according to the present invention, the inhibitory activity of the anti-inflammatory activity factors IL-6 and TNF- , And as a result, it was confirmed that the irradiation-induced chrysin significantly inhibited the production of IL-6 and TNF-a than non-irradiated chrysin (see FIG. 5).
Therefore, the anti-inflammatory activity of chrysin having a modified molecular structure by irradiation according to the present invention is increased to a maximum of __ times higher than that of untreated group, so that the physiological activity such as inflammation inhibition ability can be improved.
Therefore, the present invention can be provided as a pharmaceutical composition for the prevention or treatment of an inflammatory disease comprising, as an active ingredient, a chrysin derivative compound having a molecular structure converted by irradiation.
In particular, the term " inflammatory disease " in the present invention is a disease caused by excessive production of inflammatory cytokines TNF-a or IL-6 induced by lipopolysaccharide (LPS) or CpG-ODN stimulation, Collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, sore throat, bronchitis, pneumonia, pancreatitis, sepsis and nephritis.
The method for producing a chrysin according to the present invention comprises a step of irradiating a chrysene compound with radiation, and provides a method for producing a chrysine derivative having an anti-inflammatory activity.
The method for preparing a chrysin derivative according to the present invention may further comprise the step of dissolving the chrysin compound in alcohol and then irradiating it with radiation.
The alcohol used in the process for preparing a chrysin derivative according to the present invention may be selected from the group consisting of methanol, ethanol, n-propanol, isopropanol, ethylene glycol, polypropylene glycol and butylene glycol, Is preferably used.
The radiation used in the method for producing a chrysin derivative according to the present invention is characterized by being radiation of gamma rays, X-rays or electron beams.
The radiation used in the method for preparing a chrysin derivative according to the present invention is a gamma ray, and it is preferable to irradiate it with 15 to 50 kGy.
Accordingly, another aspect of the present invention provides a method for preparing a chrysin derivative compound that has been irradiated with a molecular structure converted.
Yet another aspect of the present invention provides a health functional food for preventing or ameliorating an inflammatory disease comprising an active ingredient of a chrysin derivative compound that is irradiated with a molecular structure and is transformed.
As used herein, the term " food " means a natural product or a processed product containing one or more nutrients, preferably a state capable of being directly eaten through a certain degree of processing, , Food, food additives, functional foods and beverages.
Foods to which a composition for preventing and / or ameliorating symptoms of an inflammatory disease according to the present invention can be added include, for example, various foods, beverages, gums, tea, vitamin complexes, and functional foods. In addition, in the present invention, the food may include special nutritive foods (e.g., crude oil, spirits, baby food, etc.), meat products, fish meat products, tofu, mackerel, noodles (Such as soy sauce, soybean paste, kochujang, mixed potatoes), sauces, confectionery (eg, snacks), candies, chocolate, gums, ice cream, milk products (eg, fermented milk, cheese, But are not limited to, pickled foods (various kinds of kimchi, pickles, etc.), beverages (e.g., fruit drinks, vegetable beverages, beverages, fermented beverages and the like) and natural seasonings (e.g. The food, beverage or food additive may be prepared by a conventional production method.
The above-mentioned " functional food " refers to a food group which is imparted with added value to function and express the function of the food by physical, biochemical or biotechnological techniques, or to control the bio-defense rhythm of the food composition, Refers to a food prepared by processing a body so as to sufficiently express the body's control function on the body, such as recovery, and the like. Specifically, it may be a health functional food. The functional food may include a food-acceptable food-aid additive, and may further comprise suitable carriers, excipients and diluents conventionally used in the production of functional foods.
In the present invention, the term " beverage " means a general term for drinking or enjoying a taste, and includes a functional beverage. The beverage is not particularly limited as long as it contains a composition for preventing and ameliorating the symptoms of inflammation and immunological diseases as an essential ingredient at the indicated ratio, and it is also possible to add various flavors or natural carbohydrates, .
Furthermore, in addition to the above-described foods, the food containing the composition for preventing or ameliorating symptoms of inflammatory diseases according to the present invention may contain flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and fillers Cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, These components can be used independently or in combination.
Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are for further illustrating the present invention, and the scope of the present invention is not limited to these examples.
Example 1 Preparation of Gamma Irradiated Chrysin
In order to prepare the gamma-irradiated chrysin of the present invention, chrysin (Sigma-Aldrich Co., St. Louis, MO, USA) used in this experiment was purchased at a purity of 97%. Chrysin was dissolved in methanol at a concentration of 1 mg / ml after quantifying 1 mg of the chrysene in the micro-tube before irradiation of the chrysene with the gamma ray. In order to irradiate the above-mentioned krysin with gamma ray, 11.1 pBq of crew and Co-60 at 10 kGy / hr at room temperature were irradiated from a gamma ray irradiation facility (IR-79, MDS Nordion Internation Ltd., Ontario, Canada) , Respectively, to obtain total absorbed doses of 15, 30 and 50 kGy in 1 mg / ml of chrysin dissolved in methanol. The absorbed doses were calculated using an alanine dosimeter (Bruker Instrument, Rheinstetten, Germanay) at 5 mm. The dosimeter system was used after standardization in accordance with the International Atomic Energy Agency (IAEA) standard, and the error of the total absorbed dose was within ± 5%. 0 kGy, which was not irradiated, was stored outside the gamma irradiation facility after obtaining the same temperature effect, and was stored in a refrigerator together with the irradiated treatment immediately after the irradiation to be used for evaluation of physiological activity (see FIG.
Example 2 HPLC Analysis of Gamma Irradiated Chrysin
The HPLC system was used to analyze the structural change of the gamma irradiation-treated chrysin prepared in Example 1 above. For the structural analysis, DAD (Diode Array Detector) detector of HPLC (Allience HPLC system, Mo. 2690, MA, USA) was used and the column was Agilent Eclipse XDB-C18 (4.6 mm × 150 mm, 5 μM) Respectively. A: B = methanol: 0.1% formic acid was used as the mobile phase, and the flow rate was 0.8 mL / min while maintaining the temperature at 35 ° C. The mobile phase was analyzed by changing the composition of MeOH (100%) to 0.1% formic acid (100%) for 45 minutes (see FIG. 1).
In order to confirm the structural change of chrysin by irradiation, the HPLC peaks of the irradiated (15, 30, 50 kGy) chrysin and the unirradiated chrysene (0 kGy) (See FIG. 2). The 50 kGy gamma - irradiated chrysin was completely absent from the peak of unchecked chrysin and confirmed the formation of gamma ray - induced new compounds CM1 and CM2. Therefore, the optimal radiation dose for securing the radiolysis - inducing compound by irradiation with chrysene was determined to be 50 kGy.
<Experimental Example 1>
Evaluation of cytotoxicity of irradiated chrysin
To assess the toxicity of gamma-irradiated chrysin to normal cells, an appropriate step to determine the appropriate concentration of the sample was carried out. Murine macrophage RAW264.7 was isolated from the Korean Cell Line Bank (KCLB) And cultured in an incubator at 37 ° C and 5% CO 2 using DMEM medium. Murine macrophage RAW264.7 cells were treated with Murine macrophage RAW264.7 cells at concentrations of 0, 3.125, 6.25, 12.5 and 25 μM. Murine macrophage RAW264.7 cells were treated with MTT assay And was assessed by law.
Cells were plated in 96-well plates, and 20 μl of MTT solution was added. The cells were incubated at 37 ° C for 2 hours. Media was removed, 200 μl of DMSO was added, and absorbance was measured at 595 nm.
As a result, RAW264.7 cells did not show cytotoxicity when treated with non-irradiated chrysin, and did not show cytotoxicity depending on the concentration when irradiated with gamma irradiation (see FIG. 3). Therefore, it was confirmed that the gamma-irradiated chrysin of the present invention did not induce apoptosis in normal cells.
< Experimental Example 2>
Irradiated Creosin The anti-inflammatory factors NO and To PEG2 Activity evaluation for
In order to observe the change of the anti-inflammatory activity according to the generation of the novel compound of the gamma-ray-treated chrysin and the gamma ray-induced new anti-inflammatory activity, the experiment of the anti-inflammatory activity change by the gamma- In order to investigate the effect of NO and PGE2, which are known to be important inflammatory factors in inflammatory reactions, 500ng / ml of lipopolysaccharide (LPS), which is an inflammation inducer, was used to measure anti-inflammatory activity of gamma-irradiated chrysin .
For the NO activity, RAW264.7 cells were cultured at a concentration of 2x10 < 5 > cells / well, and chrysin was treated for each concentration and cultured for 24 hours. After recovering the supernatant from each well, the absorbance value is evaluated at 517 nm using the Griess method (50 μL supernatant + 50 μL Griess reagent). PGE2 activity was assessed using the PGE2 detection ELISA kit in supernatants of the same conditions as above.
As a result, it was confirmed that gamma-irradiation chrysin inhibits the production of NO and PGE2 in a concentration-dependent manner in cells abnormally induced by LPS. In particular, 50 kGy gamma-irradiation chrysin showed higher inflammatory factors PGE2) (see FIG. 4). Therefore, it was confirmed that the structural transformation of chrysin prepared by using the irradiation technique improved the anti - inflammatory activity, which is the physiological activity inherent to natural products of chrysin.
< Experimental Example 3>
Irradiated Creosin Anti-inflammatory factor To Cytokine Activity evaluation for
In order to observe the change of anti-inflammatory activity according to the generation of the novel compound of gamma-ray-treated chrysin and gamma ray induction, the experiment on the change of the anti-inflammatory activity by the gamma-irradiated chrysin prepared in Example 2 is referred to as cytokine . Overexpression of the cytokine, which plays an important role in the immune system that maintains the defense system of the human body against external invasion, is the cause of the inflammatory reaction which indicates the extreme effect of the human body. Therefore, the activity of cytokine TNF-α (tumor necrosis factor-alpha) and IL-6 (interleukin-6), which are known to be important anti inflammatory factors, were evaluated.
Cytokine activity is obtained by culturing cells at a concentration of 2 × 10 5 cells / well in RAW264.7. After that, the cells are treated for each concentration of chrysin and cultured for 24 hours. The supernatant from each well was recovered and assessed using Cytokine detection ELISA kits. Absorbance values were evaluated at 450 nm.
As a result, it was confirmed that gamma-irradiation chrysin inhibits TNF-α and IL-6 secretion in a concentration-dependent manner in cells abnormally induced by LPS. In particular, 50 kGy gamma- (TNF-α and IL-6) (see FIG. 5). Therefore, it was confirmed that the structural transformation of chrysin prepared by using the irradiation technique improved the anti - inflammatory activity, which is the physiological activity inherent to the natural product of chrysin.
The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.
Claims (9)
The inflammatory disease is selected from the group consisting of inflammatory bowel disease, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, sore throat, bronchitis, pneumonia, pancreatitis, sepsis and nephritis ≪ / RTI >
The inflammatory disease is selected from the group consisting of inflammatory bowel disease, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, sore throat, bronchitis, pneumonia, pancreatitis, sepsis and nephritis ≪ / RTI >
The inflammatory disease is selected from the group consisting of inflammatory bowel disease, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, sore throat, bronchitis, pneumonia, pancreatitis, sepsis and nephritis ≪ / RTI >
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020150117307A KR101804109B1 (en) | 2015-08-20 | 2015-08-20 | Chrysin having improved antinflammation ability by irradiation, method for producing the same and method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020150117307A KR101804109B1 (en) | 2015-08-20 | 2015-08-20 | Chrysin having improved antinflammation ability by irradiation, method for producing the same and method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| KR20170022403A KR20170022403A (en) | 2017-03-02 |
| KR101804109B1 true KR101804109B1 (en) | 2017-12-04 |
Family
ID=58426743
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020150117307A KR101804109B1 (en) | 2015-08-20 | 2015-08-20 | Chrysin having improved antinflammation ability by irradiation, method for producing the same and method |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR101804109B1 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20210101761A (en) | 2020-02-11 | 2021-08-19 | 동아대학교 산학협력단 | Methods of preparing flavonoid compound using Streptomyces, and the flavonoid compound by them |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100609007B1 (en) * | 2003-12-09 | 2006-08-09 | 한국화학연구원 | Chrysin derivaties for inhibitiing selectively cox-2 enzyme |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101117072B1 (en) | 2009-05-19 | 2012-02-22 | 한국원자력연구원 | Preparation method of redginseng and blackginseng for increasing ginsenoside using radiation rays |
-
2015
- 2015-08-20 KR KR1020150117307A patent/KR101804109B1/en active IP Right Grant
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100609007B1 (en) * | 2003-12-09 | 2006-08-09 | 한국화학연구원 | Chrysin derivaties for inhibitiing selectively cox-2 enzyme |
Non-Patent Citations (1)
| Title |
|---|
| Reactivity of flavonoids with 1-hydroxyethyl radical: a γ-radiolysis study, Biochimica et Biophysica Acta, 1670(1), 28-39(2004.01.)* |
Also Published As
| Publication number | Publication date |
|---|---|
| KR20170022403A (en) | 2017-03-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Xia et al. | Nutrients and bioactive components from vinegar: A fermented and functional food | |
| JP6222626B2 (en) | Fructose absorption inhibitor | |
| KR101811122B1 (en) | A preparation method of jelly and beverage compositions containing Aronia extract reduced tannin and L-arginine | |
| KR20150111724A (en) | Beverage Compositon for Animal Companhaving Antioxidant and Deodorant | |
| KR101745338B1 (en) | Composition for preventing and treating inflammatory diseases comprising Sagassum serratifolium extract | |
| KR101872128B1 (en) | A preparation method of fermented onion composition for enhancing immune activity | |
| KR101804109B1 (en) | Chrysin having improved antinflammation ability by irradiation, method for producing the same and method | |
| KR20160141027A (en) | Phamaceutical composition or healthy food comprising water extracts from Pleurotus eryngii var. ferulea (Pf.). for treating or preventing metabolic disorder | |
| KR20140105657A (en) | A comprising an extracts of fermented Schisandra chinensis Baillon showing anti-oxidative, anti-hypertensive activity | |
| JP2008201763A (en) | Antioxidative composition | |
| KR101494664B1 (en) | fermented corni fructus composition with antioxidant activity and method of making the same | |
| KR101692889B1 (en) | Composition comprising an extract or a fraction of Daphne kamtschatica for preventing or treating inflammatory diseases | |
| WO2018155150A1 (en) | Blood uric acid level improving agent and food/drink for improving blood uric acid level | |
| KR20140145808A (en) | Preparing method of immuno-stimulating activities of polysaccharide fractions isolated and aglycon flavonoid from citrus peel | |
| JP2012006905A (en) | Composition for skin care | |
| KR101636608B1 (en) | Composition for antioxidation comprising the seed extract of cornus officinalis | |
| JP6379807B2 (en) | New resveratrol derivatives | |
| KR101854447B1 (en) | Generation of nitrite in plant extracts or grinded plant by using plasma | |
| JP6044944B2 (en) | Process for producing new processed ume products and functional composition, food composition, and pharmaceutical composition using the same | |
| JP5702514B2 (en) | Stabilization method for photodegradation of lutein | |
| KR102534736B1 (en) | Method for producing cricket powder for improving male sexual function | |
| KR101338532B1 (en) | Composition for preventing or treating colon cancer containing extract of sea cucumber | |
| Li et al. | Chemical composition and biological effects of maple syrup | |
| KR102080489B1 (en) | Method for producing buckwheat sprout with increased content of specific phenolic compounds by pectin treatment | |
| KR20120118174A (en) | Composition comprising seacucumber extract having anti-oxidation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A201 | Request for examination | ||
| E902 | Notification of reason for refusal | ||
| AMND | Amendment | ||
| E601 | Decision to refuse application | ||
| AMND | Amendment | ||
| X701 | Decision to grant (after re-examination) | ||
| GRNT | Written decision to grant |