KR20210085627A - Composition for Enhancing Immunity Comprising Complex Extracts of Gryllus bimaculatus and Tenebrio molitor Linnaues as Active Ingredient - Google Patents
Composition for Enhancing Immunity Comprising Complex Extracts of Gryllus bimaculatus and Tenebrio molitor Linnaues as Active Ingredient Download PDFInfo
- Publication number
- KR20210085627A KR20210085627A KR1020190178881A KR20190178881A KR20210085627A KR 20210085627 A KR20210085627 A KR 20210085627A KR 1020190178881 A KR1020190178881 A KR 1020190178881A KR 20190178881 A KR20190178881 A KR 20190178881A KR 20210085627 A KR20210085627 A KR 20210085627A
- Authority
- KR
- South Korea
- Prior art keywords
- extract
- cells
- cricket
- brown mealworm
- mixed
- Prior art date
- Legal status (The legal status 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 status listed.)
- Ceased
Links
- 239000000284 extract Substances 0.000 title claims abstract description 131
- 239000000203 mixture Substances 0.000 title claims abstract description 48
- 230000036039 immunity Effects 0.000 title claims abstract description 39
- 230000002708 enhancing effect Effects 0.000 title claims abstract description 26
- 239000004480 active ingredient Substances 0.000 title claims abstract description 13
- 241000238820 Gryllus bimaculatus Species 0.000 title abstract 3
- 241000254109 Tenebrio molitor Species 0.000 title abstract 3
- 210000004027 cell Anatomy 0.000 claims abstract description 56
- 210000004988 splenocyte Anatomy 0.000 claims abstract description 51
- 235000013305 food Nutrition 0.000 claims abstract description 28
- 210000001151 cytotoxic T lymphocyte Anatomy 0.000 claims abstract description 26
- 108010002350 Interleukin-2 Proteins 0.000 claims description 22
- 108060008682 Tumor Necrosis Factor Proteins 0.000 claims description 22
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 claims description 22
- 239000008194 pharmaceutical composition Substances 0.000 claims description 11
- 230000004663 cell proliferation Effects 0.000 claims description 9
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 35
- 230000001419 dependent effect Effects 0.000 abstract description 13
- 230000035755 proliferation Effects 0.000 abstract description 12
- 239000000126 substance Substances 0.000 abstract description 9
- 210000002865 immune cell Anatomy 0.000 abstract description 8
- 230000002829 reductive effect Effects 0.000 abstract description 7
- 230000035899 viability Effects 0.000 abstract description 7
- 230000024932 T cell mediated immunity Effects 0.000 abstract description 5
- 231100001083 no cytotoxicity Toxicity 0.000 abstract description 3
- 230000004913 activation Effects 0.000 abstract description 2
- 241000238814 Orthoptera Species 0.000 description 89
- 238000011282 treatment Methods 0.000 description 38
- 150000001875 compounds Chemical class 0.000 description 30
- 238000012360 testing method Methods 0.000 description 30
- 102000000588 Interleukin-2 Human genes 0.000 description 21
- 230000003833 cell viability Effects 0.000 description 21
- 239000000523 sample Substances 0.000 description 19
- 238000002156 mixing Methods 0.000 description 17
- 102000004127 Cytokines Human genes 0.000 description 15
- 108090000695 Cytokines Proteins 0.000 description 15
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 14
- 229960004397 cyclophosphamide Drugs 0.000 description 14
- 238000004458 analytical method Methods 0.000 description 13
- 235000013376 functional food Nutrition 0.000 description 12
- 230000036541 health Effects 0.000 description 12
- 230000003013 cytotoxicity Effects 0.000 description 10
- 231100000135 cytotoxicity Toxicity 0.000 description 10
- 238000002474 experimental method Methods 0.000 description 10
- 230000036737 immune function Effects 0.000 description 10
- 238000000034 method Methods 0.000 description 9
- 230000001988 toxicity Effects 0.000 description 9
- 231100000419 toxicity Toxicity 0.000 description 9
- 230000004083 survival effect Effects 0.000 description 8
- 238000005259 measurement Methods 0.000 description 7
- 210000000822 natural killer cell Anatomy 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- 230000008859 change Effects 0.000 description 6
- 230000003247 decreasing effect Effects 0.000 description 6
- 239000000654 additive Substances 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 150000003839 salts Chemical class 0.000 description 5
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 4
- 206010020751 Hypersensitivity Diseases 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
- 239000012091 fetal bovine serum Substances 0.000 description 4
- 238000009472 formulation Methods 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 239000006285 cell suspension Substances 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 235000015872 dietary supplement Nutrition 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 210000003743 erythrocyte Anatomy 0.000 description 3
- 239000000796 flavoring agent Substances 0.000 description 3
- 235000013355 food flavoring agent Nutrition 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 210000000952 spleen Anatomy 0.000 description 3
- 210000004989 spleen cell Anatomy 0.000 description 3
- SQDAZGGFXASXDW-UHFFFAOYSA-N 5-bromo-2-(trifluoromethoxy)pyridine Chemical compound FC(F)(F)OC1=CC=C(Br)C=N1 SQDAZGGFXASXDW-UHFFFAOYSA-N 0.000 description 2
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 2
- 229920001287 Chondroitin sulfate Polymers 0.000 description 2
- RGHNJXZEOKUKBD-SQOUGZDYSA-N D-gluconic acid Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O RGHNJXZEOKUKBD-SQOUGZDYSA-N 0.000 description 2
- 238000008157 ELISA kit Methods 0.000 description 2
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 244000269722 Thea sinensis Species 0.000 description 2
- 244000299461 Theobroma cacao Species 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 239000011668 ascorbic acid Substances 0.000 description 2
- 229960005070 ascorbic acid Drugs 0.000 description 2
- 235000010323 ascorbic acid Nutrition 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 229940059329 chondroitin sulfate Drugs 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 239000012636 effector Substances 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 239000000945 filler Substances 0.000 description 2
- -1 for example Chemical class 0.000 description 2
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 2
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 230000002934 lysing effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 2
- 231100000957 no side effect Toxicity 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 2
- JUJBNYBVVQSIOU-UHFFFAOYSA-M sodium;4-[2-(4-iodophenyl)-3-(4-nitrophenyl)tetrazol-2-ium-5-yl]benzene-1,3-disulfonate Chemical compound [Na+].C1=CC([N+](=O)[O-])=CC=C1N1[N+](C=2C=CC(I)=CC=2)=NC(C=2C(=CC(=CC=2)S([O-])(=O)=O)S([O-])(=O)=O)=N1 JUJBNYBVVQSIOU-UHFFFAOYSA-M 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- 229960005322 streptomycin Drugs 0.000 description 2
- 235000000346 sugar Nutrition 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000000829 suppository Substances 0.000 description 2
- 235000013616 tea Nutrition 0.000 description 2
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 description 2
- 238000002137 ultrasound extraction Methods 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 description 1
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 1
- BMYNFMYTOJXKLE-UHFFFAOYSA-N 3-azaniumyl-2-hydroxypropanoate Chemical compound NCC(O)C(O)=O BMYNFMYTOJXKLE-UHFFFAOYSA-N 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 206010049153 Allergic sinusitis Diseases 0.000 description 1
- BMFMQGXDDJALKQ-BYPYZUCNSA-N Argininic acid Chemical compound NC(N)=NCCC[C@H](O)C(O)=O BMFMQGXDDJALKQ-BYPYZUCNSA-N 0.000 description 1
- 206010003402 Arthropod sting Diseases 0.000 description 1
- 210000003771 C cell Anatomy 0.000 description 1
- 229940123150 Chelating agent Drugs 0.000 description 1
- 241000254173 Coleoptera Species 0.000 description 1
- 206010010741 Conjunctivitis Diseases 0.000 description 1
- RGHNJXZEOKUKBD-UHFFFAOYSA-N D-gluconic acid Natural products OCC(O)C(O)C(O)C(O)C(O)=O RGHNJXZEOKUKBD-UHFFFAOYSA-N 0.000 description 1
- 206010012438 Dermatitis atopic Diseases 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 1
- 206010013700 Drug hypersensitivity Diseases 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- LVGKNOAMLMIIKO-UHFFFAOYSA-N Elaidinsaeure-aethylester Natural products CCCCCCCCC=CCCCCCCCC(=O)OCC LVGKNOAMLMIIKO-UHFFFAOYSA-N 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 1
- 239000004606 Fillers/Extenders Substances 0.000 description 1
- 208000004262 Food Hypersensitivity Diseases 0.000 description 1
- 206010016946 Food allergy Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 241000238821 Gryllus Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 229920002230 Pectic acid Polymers 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 1
- 241000700157 Rattus norvegicus Species 0.000 description 1
- 241000555745 Sciuridae Species 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
- 241000254105 Tenebrio Species 0.000 description 1
- 235000005764 Theobroma cacao ssp. cacao Nutrition 0.000 description 1
- 235000005767 Theobroma cacao ssp. sphaerocarpum Nutrition 0.000 description 1
- 208000024780 Urticaria Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 229920001284 acidic polysaccharide Polymers 0.000 description 1
- 150000004805 acidic polysaccharides Chemical class 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 235000013334 alcoholic beverage Nutrition 0.000 description 1
- IAJILQKETJEXLJ-RSJOWCBRSA-N aldehydo-D-galacturonic acid Chemical compound O=C[C@H](O)[C@@H](O)[C@@H](O)[C@H](O)C(O)=O IAJILQKETJEXLJ-RSJOWCBRSA-N 0.000 description 1
- IAJILQKETJEXLJ-QTBDOELSSA-N aldehydo-D-glucuronic acid Chemical compound O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C(O)=O IAJILQKETJEXLJ-QTBDOELSSA-N 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 208000030961 allergic reaction Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 239000002543 antimycotic Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000003125 aqueous solvent Substances 0.000 description 1
- 239000007900 aqueous suspension Substances 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
- 201000008937 atopic dermatitis Diseases 0.000 description 1
- 230000001363 autoimmune Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 235000008429 bread Nutrition 0.000 description 1
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 1
- 235000014121 butter Nutrition 0.000 description 1
- 235000001046 cacaotero Nutrition 0.000 description 1
- 239000002775 capsule 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
- 150000001735 carboxylic acids Chemical class 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 238000001516 cell proliferation assay Methods 0.000 description 1
- 238000003570 cell viability assay Methods 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000019219 chocolate Nutrition 0.000 description 1
- 235000015165 citric acid Nutrition 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000010411 cooking Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 201000005311 drug allergy Diseases 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 235000006694 eating habits Nutrition 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- LVGKNOAMLMIIKO-QXMHVHEDSA-N ethyl oleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC LVGKNOAMLMIIKO-QXMHVHEDSA-N 0.000 description 1
- 229940093471 ethyl oleate Drugs 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 239000005417 food ingredient Substances 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000000174 gluconic acid Substances 0.000 description 1
- 235000012208 gluconic acid Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229940097043 glucuronic acid Drugs 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 229940093915 gynecological organic acid Drugs 0.000 description 1
- 230000003862 health status Effects 0.000 description 1
- 229920002674 hyaluronan Polymers 0.000 description 1
- 229960003160 hyaluronic acid Drugs 0.000 description 1
- 230000000521 hyperimmunizing effect Effects 0.000 description 1
- 235000015243 ice cream Nutrition 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- FZWBNHMXJMCXLU-BLAUPYHCSA-N isomaltotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)O1 FZWBNHMXJMCXLU-BLAUPYHCSA-N 0.000 description 1
- 235000015110 jellies Nutrition 0.000 description 1
- 239000008274 jelly Substances 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 229960003511 macrogol Drugs 0.000 description 1
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 1
- 239000011976 maleic acid Substances 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 239000012457 nonaqueous media Substances 0.000 description 1
- 235000012149 noodles Nutrition 0.000 description 1
- 235000003715 nutritional status Nutrition 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000003002 pH adjusting agent Substances 0.000 description 1
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 1
- 238000007911 parenteral administration Methods 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
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 235000013550 pizza Nutrition 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 239000010318 polygalacturonic acid Substances 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 238000010149 post-hoc-test Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 1
- 206010039083 rhinitis Diseases 0.000 description 1
- 229960004889 salicylic acid Drugs 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 235000013580 sausages Nutrition 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000001932 seasonal effect Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 235000011888 snacks Nutrition 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 235000014347 soups Nutrition 0.000 description 1
- 238000000551 statistical hypothesis test Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 150000003460 sulfonic acids Chemical class 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
- 235000002906 tartaric acid Nutrition 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
- A23L35/00—Foods or foodstuffs not provided for in groups A23L5/00 - A23L33/00; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
- 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
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/63—Arthropods
- A61K35/64—Insects, e.g. bees, wasps or fleas
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
-
- 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
- A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Insects & Arthropods (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Nutrition Science (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Immunology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Botany (AREA)
- Mycology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
Description
본 발명은 쌍별귀뚜라미 및 갈색거저리를 유효성분으로 포함하는 면역증진용 조성물에 관한 것으로, 더욱 구체적으로 쌍별귀뚜라미 및 갈색거저리의 혼합 추출물을 유효성분으로 포함하는 면역증진용 조성물에 관한 것이다.The present invention relates to a composition for enhancing immunity comprising paired star cricket and brown mealworm as active ingredients, and more particularly, to a composition for enhancing immunity comprising a mixed extract of paired star cricket and brown mealworm as an active ingredient.
면역은 체내에 존재하는 자기방어체계로서 인체가 외부로부터 침입해 오는 각종 물질이나 생명체를 자기 자신에 대한 이물질로 인식하여 제거하고 대사시키는 과정이다.Immunity is a self-defense system that exists in the body, and it is a process in which the human body recognizes and removes and metabolizes various substances or living things that invade from the outside as foreign substances against itself.
외부 자극에 의한 손상이나 병원 미생물의 침입으로부터 자신을 방어하기도 하지만 염증반응 등과 같이 자기 자신의 조직에 손상을 줄 수도 있다. 면역기능의 변화를 조절하여 정상으로 회복시키거나 변화의 폭을 줄여 주는 작용으로 면역기능 억제나 면역기능 증강으로 구분된다. 과민면역 반응 완화는 외부 물질에 대해 불리하게 반응하여 초래되는 알레르기 반응 자기항원 또는 변형된 자기항원에 대한 반응 등 바람직하지 않게 증가된 면역 반응을 억제시키는 것을 의미한다.It protects itself from damage caused by external stimuli or invasion of pathogenic microorganisms, but it can also damage its own tissues, such as an inflammatory response. It modulates changes in immune function to restore normalcy or reduces the range of changes, and is classified as suppression of immune function or enhancement of immune function. Alleviation of hyperimmune response means suppressing an undesirably increased immune response, such as a response to an allergic reaction self-antigen or a modified self-antigen resulting from an adverse reaction to a foreign substance.
면역 기능에 영향을 미치는 요인에는 유전적, 환경적 요인, 나이, 성별, 심리적 스트레스, 영양상태, 식이습관, 질병 등이 있다.Factors that affect immune function include genetic and environmental factors, age, gender, psychological stress, nutritional status, dietary habits, and disease.
면역 기능의 조절 기능성을 갖는 건강기능식품은 현재까지, 저하된 면역기능을 증진 시키는 기능성과 과도한 면역기능을 조절하는 기능성, 자기 성분에 대한 면역반응을 조절하는 자가면역기능 조절 기능성으로 구분된다. 면역기능이 저하되면 과민반응을 일으키는 경우가 있는데 이러한 과민반응은 천식, 계절성 또는 동년성 비염, 알러지성 부비강염, 결막염, 식품과 약품에 의한 알러지, 아토피성 피부염, 두드러기, 벌침에 의한 알러지 등과 같이 다양하게 나타난다.So far, health functional foods with the function of regulating the immune function are divided into the function of enhancing the reduced immune function, the function of regulating the excessive immune function, and the function of regulating the autoimmune function of controlling the immune response to the self component. When the immune function is lowered, hypersensitivity reactions may occur. Such hypersensitivity reactions are diverse, such as asthma, seasonal or same-year rhinitis, allergic sinusitis, conjunctivitis, food and drug allergy, atopic dermatitis, urticaria, and allergy to bee stings. appear to be
이러한 여러 원인으로 인해 면역기능에 장애가 발생하는 문제점을 극복하기 위하여, 다양한 면역 증강제 및 치료제가 사용되고 있으나, 부작용 문제들이 있고, 주로 시판되는 것은 예방보다는 치료용으로 복용되는 것으로, 부작용이 없어 장기 복용이 가능하고, 예방용으로도 적합한 면역증진용 조성물이 요구되는 실정이다.In order to overcome the problem that immune function is impaired due to these various causes, various immune enhancers and therapeutic agents are used, but there are side effects, and those on the market are mainly taken for treatment rather than prevention, and long-term use is not possible because there are no side effects. There is a need for a composition for enhancing immunity that is possible and suitable for prophylaxis.
이에, 본 발명자들은 면역관련 기전을 바탕으로 면역증진 기능을 갖는 물질을 스크리닝하던 중 우수한 효능이 있는 소재로서 쌍별귀뚜라미와 갈색거저리를 선별하였으며, 쌍별귀뚜라미와 갈색거저리의 혼합물이 세포 독성이 없고, 비장세포, NK(Natural Killer) 세포, 및 CTL(Cytotoxic T Lymphocyte) 세포의 세포 증식을 촉진시켜 세포성 면역 반응을 증진시키고, 사이토카인인 TNF-α 및 IL-2의 발현을 증가시켜 우수한 면역증진 효과가 있는 것을 확인하고 본 발명을 완성하였다. Accordingly, the present inventors selected paired crickets and brown mealworms as materials with excellent efficacy while screening for substances having an immune enhancing function based on an immune-related mechanism, and the mixture of paired star crickets and brown mealworms has no cytotoxicity, and the spleen It promotes cell proliferation of cells, NK (Natural Killer) cells, and CTL (Cytotoxic T Lymphocyte) cells to enhance a cellular immune response, and increases the expression of cytokines TNF-α and IL-2 to enhance immunity It was confirmed that there is and completed the present invention.
따라서, 본 발명의 주된 목적은 면역증진에 뛰어난 효과를 나타내는 쌍별귀뚜라미와 갈색거저리의 혼합물을 유효성분으로 포함하는 면역증진용 조성물을 제공하는 데 있다.Therefore, the main object of the present invention is to provide a composition for enhancing immunity comprising a mixture of crickets and brown mealworm as an active ingredient, which exhibits an excellent effect in enhancing immunity.
본 발명의 다른 목적 및 이점은 하기의 발명의 상세한 설명, 청구범위 및 도면에 의해 보다 명확하게 된다.Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.
본 발명의 한 양태에 따르면, 본 발명은 쌍별귀뚜라미 및 갈색거저리의 혼합 추출물을 유효성분으로 포함하는 면역증진용 식품 조성물을 제공한다.According to one aspect of the present invention, the present invention provides a food composition for enhancing immunity comprising a mixed extract of paired star cricket and brown mealworm as an active ingredient.
본 발명의 용어 "쌍별귀뚜라미(Gryllus bimaculatus)"는 메뚜기목 귀뚜라미과의 곤충을 의미한다. 또한, 본 발명의 "갈색거저리(Tenebrio molitor Linnaues)"는 딱정벌레목 거저리과의 곤충의 유충을 의미한다. 대한민국 식품의약품안전처는 한시적 식품원료로 인정받은 갈색거저리 유충과 쌍별귀뚜라미를 모든 영업자가 식품의 제조·가공·조리에 사용할 수 있도록 허용한바 있다. 상기 쌍별귀뚜라미 및 갈색거저리는 농림축산식품부에서 식품원료로 등록된 이후 명칭을 '쌍별이'로, 갈색거저리 유충은 '고소애'로 지칭된 바 있으며, 본 발명에서는 이들 용어 상호간에 동일한 의미로 사용한다. The term "double star cricket ( Gryllus)" in the present invention bimaculatus )" means an insect of the locustaceae family Acridae. In addition, the "brown mealworm ( Tenebrio)" of the present invention molitor Linnaues) means the larvae of insects of the Coleoptera family. The Ministry of Food and Drug Safety of the Republic of Korea permits all business operators to use brown mealworm larvae and twin-starred crickets, which have been recognized as temporary food ingredients, for manufacturing, processing and cooking of food. After being registered as food raw materials by the Ministry of Agriculture, Food and Rural Affairs, the pair-byeol cricket and brown mealworm have been called 'ssangbyeol' and the brown mealworm larvae have been called 'gosoae', and in the present invention, these terms are identical to each other. used in meaning
본 발명에서 사용되는 용어, "추출물"은 천연물의 추출 처리에 의하여 얻어지는 추출액, 상기 추출액의 희석액이나 농축액, 상기 추출액을 건조하여 얻어지는 건조물, 상기 추출액의 조정제물이나 정제물, 또는 이들의 혼합물 등, 추출액 자체 및 추출액을 이용하여 형성 가능한 모든 제형의 추출물을 포함한다. 본 발명의 상기 추출물은 바람직하게 추출 후 건조 분말 형태로 제조되어 사용될 수 있다.As used herein, the term "extract" refers to an extract obtained by extraction of a natural product, a diluted or concentrated liquid of the extract, a dried product obtained by drying the extract, a prepared or purified product of the extract, or a mixture thereof, etc., It includes extracts of all formulations that can be formed using the extract itself and the extract. The extract of the present invention may be prepared and used in the form of a dry powder after extraction.
본 발명의 상기 추출에 있어서, 상기 추출물을 추출하는 방법은 특별히 제한되지 아니하며, 당해 기술 분야에서 통상적으로 사용하는 방법에 따라 추출할 수 있다. 상기 추출 방법의 비제한적인 예로는, 초음파 추출법, 용매분획법, 원심분리법 등을 들 수 있으며, 이들은 단독으로 수행되거나 2종 이상의 방법을 병용하여 수행될 수 있다. In the extraction of the present invention, the method of extracting the extract is not particularly limited, and may be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method include an ultrasonic extraction method, a solvent fractionation method, a centrifugation method, and the like, and these may be performed alone or in combination of two or more methods.
본 발명에서 용어, "면역"은 체내에 존재하는 자기방어체계로서 인체가 외부로부터 침입해 오는 각종 물질이나 생명체를 자기 자신에 대한 이물질로 인식하여 제거하고 대사시키는 과정을 의미하며, "면역증진"은 면역기능을 증진시키는 것을 의미한다. As used herein, the term “immunity” refers to the process of recognizing, removing and metabolizing various substances or living organisms that invade from the outside as a self-defense system existing in the body, as foreign substances to itself, and “improving immunity” means to enhance the immune function.
또한, 본 명세서에서 "유효성분"이란 단독으로 목적하는 활성을 나타내거나, 또는 그 자체는 활성이 없는 담체와 함께 활성을 나타낼 수 있는 성분을 의미한다.In addition, as used herein, the term "active ingredient" refers to a component that alone exhibits the desired activity, or can exhibit activity together with a carrier having no activity by itself.
본 발명의 면역증진용 식품 조성물에서, 상기 쌍별귀뚜라미와 상기 갈색거저리는 이에 제한되는 것은 아니나 바람직하게는 70-80:20-30 중량부로 혼합될 수 있고, 가장 바람직하게는 75:25의 중량부로 혼합될 수 있으며, 면역증진에 있어서 뚜렷하게 개선된 시너지 효과를 내는 측면에서 상기 비율로 혼합되는 것이 가장 바람직하다.In the food composition for enhancing immunity of the present invention, the paired cricket and the brown mealworm are not limited thereto, but preferably 70-80:20-30 parts by weight, and most preferably 75:25 parts by weight. It can be mixed, and it is most preferable to mix in the above ratio in terms of producing a distinctly improved synergistic effect in enhancing immunity.
본 발명의 면역증진용 식품 조성물에서, 상기 조성물은 비장세포, NK(Natural Killer) 세포, 및 CTL(Cytotoxic T Lymphocyte) 세포의 세포 증식을 증진하는 것을 특징으로 한다. 구체적으로 본 발명의 바람직한 실시예에서는 상기 쌍별귀뚜라미 및 갈색거저리의 추출물 또는 초음파 추출물의 혼합물이 Cy(cyclophosphamide) 화합물을 처리하여 면역력이 낮아진 비장세포에서 농도 의존적으로 비장세포의 생존율과 증식율을 개선시키고(도 3 내지 도 5 참조), NK(Natural Killer) 세포인 AR42J 세포에서 농도 의존적으로 NK 세포의 생존율과 증식율을 개선시키고(도 10 내지 도 13 참조), CTL(Cytotoxic T Lymphocyte) 세포인 HL-60 세포에서 농도 의존적으로 CTL 세포의 생존율과 증식율을 개선시켜 저하된 면역세포의 활성을 실질적으로 증가시켜주고, 이에 따라 전반적으로 세포성 면역 반응 체계의 활성을 통해 면역력을 직접적으로 개선하는 효과를 확인할 수 있었다(도 14 및 도 15 참조). In the food composition for enhancing immunity of the present invention, the composition is characterized in that it promotes cell proliferation of splenocytes, NK (Natural Killer) cells, and CTL (Cytotoxic T Lymphocyte) cells. Specifically, in a preferred embodiment of the present invention, the mixture of extracts or ultrasonic extracts of crickets and brown mealworms is treated with a Cy (cyclophosphamide) compound to improve the survival rate and proliferation rate of splenocytes in a concentration-dependent manner in splenocytes with reduced immunity ( 3 to 5), improves the survival rate and proliferation rate of NK cells in a concentration-dependent manner in AR42J cells, which are Natural Killer (NK) cells (see FIGS. 10 to 13), and HL-60, which is a Cytotoxic T Lymphocyte (CTL) cell. By improving the survival rate and proliferation rate of CTL cells in a concentration-dependent manner in the cell, it substantially increases the activity of the reduced immune cells, and thus the effect of directly improving immunity through the activity of the cellular immune response system in general can be confirmed. (see FIGS. 14 and 15 ).
본 발명의 면역증진용 식품 조성물에서, 상기 조성물은 TNF-α 및 IL-2의 발현을 증가시키는 것을 특징으로 한다. 구체적으로 본 발명의 바람직한 실시예에서는 상기 쌍별귀뚜라미 및 갈색거저리의 추출물 또는 초음파 추출물의 혼합물이 Cy 화합물의 처리로 저하된 사이토카인(TNF-α, IL-2)의 발현을 실질적으로 증가시켜주고, 이에 따라 전반적으로 면역력을 개선하는 효과를 나타내는 것을 확인할 수 있었다(도 6 내지 도 9 참조). In the food composition for enhancing immunity of the present invention, the composition is characterized in that it increases the expression of TNF-α and IL-2. Specifically, in a preferred embodiment of the present invention, the mixture of extracts or ultrasonic extracts of the paired cricket and brown mealworm substantially increases the expression of cytokines (TNF-α, IL-2) lowered by the treatment of the Cy compound, Accordingly, it was confirmed that the effect of improving the overall immunity was exhibited (see FIGS. 6 to 9 ).
상기 식품 조성물은 건강기능식품의 형태로 사용될 수 있으나, 이에 제한되는 것은 아니다.The food composition may be used in the form of health functional food, but is not limited thereto.
또한, 상기 식품 조성물은 유효성분 이외에 식품학적으로 허용 가능한 식품보조첨가제를 포함할 수 있다.In addition, the food composition may include a food pharmaceutically acceptable food supplement additive in addition to the active ingredient.
본 발명에 있어서, "식품보조첨가제"란 식품에 보조적으로 첨가될 수 있는 구성요소를 의미하며, 각 제형의 건강기능식품을 제조하는데 첨가되는 것으로서 당업자가 적절히 선택하여 사용할 수 있다. 식품보조첨가제의 예로는 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 충진제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등이 포함되지만, 상기 예들에 의해 본 발명의 식품보조 첨가제의 종류가 제한되는 것은 아니다.In the present invention, "food supplementary additive" means a component that can be added to food as an auxiliary, added to the manufacture of health functional food of each formulation can be appropriately selected and used by those skilled in the art. Examples of food supplement additives include various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners , pH adjuster, stabilizer, preservative, glycerin, alcohol, carbonation agent used in carbonated beverages, etc., but the types of food supplement additives of the present invention are not limited by the above examples.
본 발명의 식품 조성물에는 건강기능식품이 포함될 수 있다. 본 발명에 있어서, "건강기능식품"이란 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 정제, 캅셀, 분말, 과립, 액상 및 환 등의 형태로 제조 및 가공한 식품을 말한다. 여기서 “기능성”이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻는 것을 의미한다. 본 발명의 건강기능식품은 당업계에서 통상적으로 사용되는 방법으로 제조 가능하며, 상기 제조시에는 당업계에서 통상적으로 첨가하는 원료 및 성분을 첨가하여 제조할 수 있다. The food composition of the present invention may include a health functional food. In the present invention, "health functional food" refers to a food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. using raw materials or ingredients useful for the human body. Here, the term “functionality” refers to obtaining useful effects for health purposes, such as regulating nutrients or physiological actions with respect to the structure and function of the human body. The health functional food of the present invention can be prepared by a method commonly used in the art, and at the time of manufacture, it can be prepared by adding raw materials and components commonly added in the art.
또한, 상기 건강기능식품의 제형 또한 건강기능식품으로 인정되는 제형이면 제한 없이 제조될 수 있다. 본 발명의 식품 조성물은 다양한 형태의 제형으로 제조될 수 있으며, 일반 약품과는 달리 식품을 원료로 하여 약품의 장기 복용 시 발생할 수 있는 부작용 등이 없는 장점이 있고, 휴대성이 뛰어나, 본 발명의 건강기능식품은 면역증진을 위한 보조제로 섭취가 가능하다.In addition, the dosage form of the health functional food may also be manufactured without limitation as long as it is a dosage form recognized as a health functional food. The food composition of the present invention can be prepared in various forms, and unlike general drugs, it has the advantage that there are no side effects that may occur during long-term administration of the drug using food as a raw material, and has excellent portability, Health functional food can be consumed as an adjuvant for immune enhancement.
본 발명의 건강기능식품이 취할 수 있는 형태에는 제한이 없으며, 통상적인 의미의 식품을 모두 포함할 수 있고, 기능성 식품 등 당업계에 알려진 용어와 혼용이 가능하다. 아울러 본 발명의 건강기능식품은 당업자의 선택에 따라 식품에 포함될 수 있는 적절한 기타 보조 성분과 공지의 첨가제를 혼합하여 제조할 수 있다. 첨가할 수 있는 식품의 예로는 육류, 소세지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알코올 음료 및 비타민 복합제 등이 있으며, 본 발명에 따른 화학식 1로 표시되는 화합물을 주성분으로 하여 제조한 즙, 차, 젤리 및 주스 등에 첨가하여 제조할 수 있다. 또한, 동물을 위한 사료로 이용되는 식품도 포함된다.There is no limitation on the form that the health functional food of the present invention can take, and it may include any food in a conventional sense, and may be used interchangeably with terms known in the art, such as functional food. In addition, the health functional food of the present invention can be prepared by mixing known additives with other suitable auxiliary ingredients that may be included in the food according to the selection of those skilled in the art. Examples of foods that can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages and There are vitamin complexes and the like, and it can be prepared by adding the compound represented by Formula 1 according to the present invention as a main component to juice, tea, jelly, juice, and the like. Also included are foods used as feed for animals.
본 발명의 다른 양태에 따르면, 본 발명은 쌍별귀뚜라미 및 갈색거저리의 혼합 추출물을 유효성분으로 포함하는 면역증진용 약학적 조성물을 제공한다.According to another aspect of the present invention, the present invention provides a pharmaceutical composition for enhancing immunity comprising a mixed extract of paired star cricket and brown mealworm as an active ingredient.
본 발명의 면역증진용 약학적 조성물은 비장세포, NK(Natural Killer) 세포, 및 CTL(Cytotoxic T Lymphocyte) 세포의 세포 증식을 증진하는 것을 특징으로 한다. 또한, 본 발명의 면역증진용 약학적 조성물은 사이토카인인 TNF-α 및 IL-2의 발현을 증가시키는 것을 특징으로 한다. "면역증진"의 의미는 상기에서 설명한 바와 같다.The pharmaceutical composition for enhancing immunity of the present invention is characterized in that it promotes cell proliferation of splenocytes, NK (Natural Killer) cells, and CTL (Cytotoxic T Lymphocyte) cells. In addition, the pharmaceutical composition for enhancing immunity of the present invention is characterized in that it increases the expression of cytokines TNF-α and IL-2. The meaning of "immune enhancement" is the same as described above.
한편, 본 발명의 쌍별귀뚜라미 및 갈색거저리의 혼합물을 유효성분으로 포함하는 면역증진용 약학적 조성물은 약학적으로 허용 가능한 염의 형태로 제조될 수 있다. On the other hand, the pharmaceutical composition for enhancing immunity comprising a mixture of the pair-star cricket and brown mealworm of the present invention as an active ingredient may be prepared in the form of a pharmaceutically acceptable salt.
구체적으로 산을 첨가함으로써 염을 형성할 수 있고, 예를 들어 무기산(예: 염산, 히드로브롬산, 인산, 질산, 황산 등), 유기 카르복실산(예: 아세트산, 트리플루오로아세트산과 같은 할로 아세트산, 프로피온산, 말레산, 숙신산, 말산, 시트르산, 타르타르산, 살리실산), 및 산성 당(글루쿠론산, 갈락투론산, 글루콘산, 아스코르브산), 산성 폴리사카리드(예: 히알우론산, 콘드로이틴 술페이트, 아르기닌산), 콘드로이틴 술페이트와 같은 술폰산 당 에스테르를 포함하는 유기 술폰산(예: 메탄, 술폰산, p-톨루엔 술폰산) 등을 첨가하여 염을 형성할 수 있다.Specifically, salts can be formed by addition of acids, for example, inorganic acids (e.g. hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, sulfuric acid, etc.), organic carboxylic acids (e.g. acetic acid, trifluoroacetic acid, etc.) acetic acid, propionic acid, maleic acid, succinic acid, malic acid, citric acid, tartaric acid, salicylic acid), and acidic sugars (glucuronic acid, galacturonic acid, gluconic acid, ascorbic acid), acidic polysaccharides such as hyaluronic acid, chondroitin sulfate, arginic acid), organic sulfonic acids including sulfonic acid sugar esters such as chondroitin sulfate (eg, methane, sulfonic acid, p-toluene sulfonic acid), etc. may be added to form salts.
본 발명의 약학적 조성물은 세포 독성이 없는 천연 추출물에서 분리되는 물질들로 이루어져 임상투여시 경구 또는 비경구로 투여 가능하며, 일반적인 의약품 제제의 형태로 사용될 수 있다.The pharmaceutical composition of the present invention is composed of substances separated from natural extracts without cytotoxicity and can be administered orally or parenterally during clinical administration, and can be used in the form of general pharmaceutical preparations.
즉, 본 발명의 약학적 조성물은 실제로 경구 또는 비경구의 여러 가지 제형으로 투여될 수 있는데, 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 비경구투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조제제, 좌제가 포함된다. 비수성용제, 현탁용제로는 프로필렌글리콜(Propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 리우린지, 글리세로제라틴 등이 사용될 수 있다.That is, the pharmaceutical composition of the present invention can actually be administered in various oral or parenteral formulations, and when formulated, commonly used fillers, extenders, binders, wetting agents, disintegrants, diluents such as surfactants or excipients are used. is prepared by Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base of the suppository, witepsol, macrogol, tween 61, cacao butter, liulinji, glycerogelatin, and the like can be used.
또한, 본 발명의 약학적 조성물은 생리식염수 또는 유기용매와 같이 약제로 허용된 여러 전달체(carrier)와 혼합하여 사용될 수 있고, 안정성이나 흡수성을 증가시키기 위하여 글루코스, 수크로스 또는 덱스트란과 같은 카보하이드레이트, 아스코르브 산(ascorbic acid) 또는 글루타치온과 같은 항산화제(antioxidants), 킬레이트화제(chelating agents), 저분자 단백질 또는 다른 안정화제(stabilizers)들이 약제로 사용될 수 있다.In addition, the pharmaceutical composition of the present invention can be used by mixing with various pharmaceutically acceptable carriers such as physiological saline or organic solvents, and carbohydrates such as glucose, sucrose or dextran to increase stability or absorption , antioxidants such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers may be used as pharmaceuticals.
본 발명의 약학적 조성물의 유효용량은 0.01㎎/㎏ 내지 10㎎/㎏이고, 바람직하게는 0.1 내지 1㎎/㎏ 이며, 하루 1회 내지 3회 투여될 수 있다.The effective dose of the pharmaceutical composition of the present invention is 0.01 mg/kg to 10 mg/kg, preferably 0.1 to 1 mg/kg, and may be administered 1 to 3 times a day.
본 발명의 약학적 조성물의 총 유효량은 볼루스(bolus) 형태 혹은 상대적으로 짧은 기간 동안 주입(infusion) 등에 의해 단일 투여량(single does)으로 환자에게 투여될 수 있으며, 다중 투여량(multiple does)이 장기간 투여되는 분할 치료 방법(fractionated treatment protocol)에 의해 투여될 수 있다. 상기 농도는 약의 투여 경로 및 치료 횟수뿐만 아니라 환자의 나이 및 건강상태 등 다양한 요인들을 고려하여 환자의 유효 투여량이 결정되는 것이므로 이러한 점을 고려할 때, 이 분야의 통상적인 지식을 가진 자라면 본 발명의 약학적 조성물로서의 특정한 용도에 따른 적절한 유효 투여량을 결정할 수 있을 것이다.The total effective amount of the pharmaceutical composition of the present invention may be administered to a patient in the form of a bolus or by infusion for a relatively short period of time as a single dose, and multiple doses may be used. It can be administered by this long administered fractionated treatment protocol. Since the concentration is determined by considering various factors such as the age and health status of the patient as well as the administration route and number of treatments, the effective dosage of the patient is determined. It will be possible to determine an appropriate effective dosage according to a particular use as a pharmaceutical composition.
본 발명의 특징 및 이점을 요약하면 다음과 같다:The features and advantages of the present invention are summarized as follows:
(a) 본 발명은 쌍별귀뚜라미 및 갈색거저리의 추출물 또는 초음파 추출물을 유효성분으로 포함하는 면역증진용 조성물을 제공한다.(a) The present invention provides a composition for enhancing immunity comprising an extract or ultrasonic extract of twin-star cricket and brown mealworm as an active ingredient.
(b) 본 발명의 면역증진용 조성물은 세포 독성이 없고, 농도 의존적으로 비장세포, NK(Natural Killer) 세포, 및 CTL(Cytotoxic T Lymphocyte) 세포의 생존율과 증식율을 개선시켜 저하된 면역세포의 활성을 실질적으로 증가시켜주고, 이에 따라 전반적으로 세포성 면역 반응 체계의 활성을 통해 면역력을 직접적으로 개선함으로써 우수한 면역증진 효과가 있다. (b) the composition for enhancing immunity of the present invention has no cytotoxicity, and improves the viability and proliferation rate of splenocytes, NK (Natural Killer) cells, and CTL (Cytotoxic T Lymphocyte) cells in a concentration-dependent manner, thereby reducing the activity of immune cells , and thus has an excellent immune-promoting effect by directly improving immunity through activation of the overall cellular immune response system.
(c) 또한, 본 발명의 면역증진용 조성물은 천연에서 존재하는 식용물질인 쌍별귀뚜라미와 갈색 거저리를 배합하여 인체에 매우 안전할 뿐만 아니라, 안정성도 매우 탁월하여 식품, 및 의약품 분야에서 면역증진의 목적으로 유용하게 이용 가능하다. (c) In addition, the composition for enhancing immunity of the present invention is not only very safe for the human body by blending edible substances that exist in nature, such as twin-star cricket and brown mealworm, but also very excellent in stability, so that it can be used to enhance immunity in food and pharmaceutical fields. useful for the purpose.
도 1은 쌍별귀뚜라미(A)와 갈색거저리(B) 추출물 처리에 따른 비장세포의 세포 생존율을 측정하여 나타낸 그래프이다(시험군별 평균값의 통계적 유의수준은 p<0.05에 대한 각각의 부집단으로 표기).
도 2는 쌍별귀뚜라미(A)와 갈색거저리(B) 초음파 추출물 처리에 따른 비장세포의 세포 생존율을 측정하여 나타낸 그래프이다(시험군별 평균값의 통계적 유의수준은 p<0.05에 대한 각각의 부집단으로 표기).
도 3은 쌍별귀뚜라미와 갈색거저리의 혼합 추출물(A) 및 혼합 초음파 추출물(B) 처리에 따른 비장세포의 세포 생존율을 측정하여 나타낸 그래프이다(시험군별 평균값의 통계적 유의수준은 p<0.05에 대한 각각의 부집단으로 표기).
도 4는 Cy(cyclophosphamide) 화합물을 처리한 비장세포에서 쌍별귀뚜라미와 갈색거저리의 혼합 추출물(A) 및 혼합 초음파 추출물(B)의 비율별 처리에 따른 비장세포의 세포 생존율을 측정하여 나타낸 그래프이다(시험군별 평균값의 통계적 유의수준은 p<0.05에 대한 각각의 부집단으로 표기).
도 5는 Cy(cyclophosphamide) 화합물을 처리한 비장세포에서 쌍별귀뚜라미와 갈색거저리를 75:25 중량비로 혼합한 추출물(A) 및 초음파 추출물(B)의 처리에 따른 비장세포의 세포 생존율을 측정하여 나타낸 그래프이다(시험군별 평균값의 통계적 유의수준은 p<0.05에 대한 각각의 부집단으로 표기).
도 6은 Cy(cyclophosphamide) 화합물을 처리한 비장세포에서 쌍별귀뚜라미와 갈색거저리의 추출물의 혼합 비율에 따른 비장세포의 사이토카인(TNF-α, IL-2) 발현량 변화를 측정하여 나타낸 그래프이다(시험군별 평균값의 통계적 유의수준은 p<0.05에 대한 각각의 부집단으로 표기).
도 7은 Cy(cyclophosphamide) 화합물을 처리한 비장세포에서 쌍별귀뚜라미와 갈색거저리의 초음파 추출물의 혼합 비율에 따른 비장세포의 사이토카인(TNF-α, IL-2) 발현량 변화를 측정하여 나타낸 그래프이다(시험군별 평균값의 통계적 유의수준은 p<0.05에 대한 각각의 부집단으로 표기).
도 8은 Cy(cyclophosphamide) 화합물을 처리한 비장세포에서 쌍별귀뚜라미와 갈색거저리를 75:25 중량비로 혼합한 추출물의 농도별 처리에 따른 비장세포의 사이토카인(TNF-α, IL-2) 발현량 변화를 측정하여 나타낸 그래프이다(시험군별 평균값의 통계적 유의수준은 p<0.05에 대한 각각의 부집단으로 표기).
도 9는 Cy(cyclophosphamide) 화합물을 처리한 비장세포에서 쌍별귀뚜라미와 갈색거저리를 75:25 중량비로 혼합한 초음파 추출물의 농도별 처리에 따른 비장세포의 사이토카인(TNF-α, IL-2) 발현량 변화를 측정하여 나타낸 그래프이다(시험군별 평균값의 통계적 유의수준은 p<0.05에 대한 각각의 부집단으로 표기).
도 10은 쌍별귀뚜라미(A)와 갈색거저리(B) 추출물 처리에 따른 AR42J 세포의 세포 생존율을 측정하여 나타낸 그래프이다(시험군별 평균값의 통계적 유의수준은 p<0.05에 대한 각각의 부집단으로 표기).
도 11은 쌍별귀뚜라미(A)와 갈색거저리(B)의 초음파 추출물 처리에 따른 AR42J 세포의 세포 생존율을 측정하여 나타낸 그래프이다(시험군별 평균값의 통계적 유의수준은 p<0.05에 대한 각각의 부집단으로 표기).
도 12는 쌍별귀뚜라미와 갈색거저리 혼합 추출물(A) 및 혼합 초음파 추출물(B)의 비율별 처리에 따른 AR42J 세포의 세포 생존율을 측정하여 나타낸 그래프이다(시험군별 평균값의 통계적 유의수준은 p<0.05에 대한 각각의 부집단으로 표기).
도 13은 쌍별귀뚜라미와 갈색거저리의 추출물 또는 초음파 추출물을 75:25의 중량비로 혼합한 혼합 추출물(A)과 혼합 초음파 추출물(B)의 처리에 따른 AR42J 세포의 세포 생존율을 측정하여 나타낸 그래프이다(시험군별 평균값의 통계적 유의수준은 p<0.05에 대한 각각의 부집단으로 표기).
도 14는 쌍별귀뚜라미와 갈색거저리를 75:25의 중량비로 혼합한 혼합 추출물(A)과 혼합 초음파 추출물(B)의 처리에 따른 CTL(Cytotoxic T Lymphocyte) 세포인 HL-60 세포의 세포 생존율을 측정하여 나타낸 그래프이다(시험군별 평균값의 통계적 유의수준은 p<0.05에 대한 각각의 부집단으로 표기).
도 15는 쌍별귀뚜라미와 갈색거저리를 75:25의 중량비로 혼합한 혼합 추출물(A)과 혼합 초음파 추출물(B)을 비장세포에 HL-60 세포와 함께 처리하고 HL-60 세포의 세포 생존율을 측정하여 나타낸 그래프이다(시험군별 평균값의 통계적 유의수준은 p<0.05에 대한 각각의 부집단으로 표기).1 is a graph showing the measurement of cell viability of splenocytes according to pairwise cricket (A) and brown mealworm (B) extract treatment (the statistical significance level of the mean value for each test group is expressed as each subgroup for p <0.05) .
2 is a graph showing the cell viability of splenocytes according to the ultrasonic extract treatment of paired cricket (A) and brown mealworm (B) (the statistical significance level of the average value for each test group is expressed as each subgroup for p <0.05). ).
3 is a graph showing the cell viability of splenocytes according to the treatment of the mixed extract (A) and the mixed ultrasonic extract (B) of pairwise cricket and brown mealworm (the statistical significance level of the mean value for each test group is p <0.05, respectively. denoted as a subgroup of ).
4 is a graph showing the measurement of cell viability of splenocytes according to the ratio of the mixed extract (A) and the mixed ultrasonic extract (B) of paired cricket and brown mealworm in splenocytes treated with Cy (cyclophosphamide) compound (B). The statistical significance level of the mean value for each test group is expressed as each subgroup for p<0.05).
5 is a Cy (cyclophosphamide) compound-treated splenocytes in splenocytes treated with an extract (A) and an ultrasonic extract (B) in a 75:25 weight ratio of twin-star crickets and brown mealworm by measuring the cell viability of splenocytes. It is a graph (statistical significance level of the mean value for each test group is expressed as each subgroup for p<0.05).
Figure 6 is a graph showing the change in cytokine (TNF-α, IL-2) expression in splenocytes according to the mixing ratio of extracts of paired cricket and brown mealworm in splenocytes treated with Cy (cyclophosphamide) compound ( The statistical significance level of the mean value for each test group is expressed as each subgroup for p<0.05).
7 is a graph showing the change in cytokine (TNF-α, IL-2) expression in splenocytes according to the mixing ratio of the ultrasonic extracts of twin-star cricket and brown mealworm in splenocytes treated with Cy (cyclophosphamide) compound. (The statistical significance level of the mean value for each test group is indicated by each subgroup for p<0.05).
Figure 8 shows the cytokine (TNF-α, IL-2) expression level of splenocytes according to the concentration-specific treatment of an extract obtained by mixing paired cricket and brown mealworm in a 75:25 weight ratio in splenocytes treated with Cy (cyclophosphamide) compound; This is a graph showing the measured change (the statistical significance level of the mean value for each test group is indicated by each subgroup for p<0.05).
9 shows cytokine (TNF-α, IL-2) expression in splenocytes according to concentration-specific treatment of an ultrasonic extract obtained by mixing paired cricket and brown mealworm in a 75:25 weight ratio in splenocytes treated with Cy (cyclophosphamide) compound; This is a graph showing the change in dose (the statistical significance level of the mean value for each test group is indicated by each subgroup for p<0.05).
10 is a graph showing the measurement of the cell viability of AR42J cells according to the pairwise cricket (A) and brown mealworm (B) extract treatment (the statistical significance level of the average value for each test group is expressed as each subgroup for p <0.05) .
11 is a graph showing the measurement of the cell viability of AR42J cells according to the ultrasonic extract treatment of paired crickets (A) and brown mealworm (B) (the statistical significance level of the mean value for each test group is p <0.05 for each subgroup. Mark).
12 is a graph showing the measurement of the cell viability of AR42J cells according to the ratio treatment of the pairwise cricket and brown mealworm mixed extract (A) and the mixed ultrasonic extract (B) (the statistical significance level of the mean value for each test group is p <0.05) denoted by each subgroup).
13 is a graph showing the measurement of the cell viability of AR42J cells according to the treatment of the mixed extract (A) and the mixed ultrasonic extract (B) in which an extract or ultrasonic extract of paired cricket and brown mealworm was mixed in a weight ratio of 75:25 ( The statistical significance level of the mean value for each test group is expressed as each subgroup for p<0.05).
14 shows the cell viability of CTL (Cytotoxic T Lymphocyte) cells, HL-60 cells, according to the treatment of the mixed extract (A) and the mixed ultrasonic extract (B) in which the pairwise cricket and brown mealworm are mixed in a weight ratio of 75:25. (The statistical significance level of the mean value for each test group is expressed as each subgroup for p<0.05).
15 is a mixed extract (A) and a mixed ultrasonic extract (B) mixed with paired crickets and brown mealworm in a weight ratio of 75:25, treated with HL-60 cells in splenocytes, and the cell viability of HL-60 cells is measured. (The statistical significance level of the mean value for each test group is expressed as each subgroup for p<0.05).
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하기로 한다. 이들 실시예는 단지 본 발명을 예시하기 위한 것이므로, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는다.Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and therefore, the scope of the present invention should not be construed as being limited by these examples.
[실험재료 및 방법][Test materials and methods]
1-1. 세포 배양1-1. cell culture
비장세포(Splenocytes)는 Wistar Rat의 비장을 적출 후 핀셋과 메쉬를 이용하여 단일세포 부유액을 제조하였다. 단일세포 부유액을 RPMI-1640 배양액으로 3회 원심 침전(1,000g, 5 min, 4℃)하여 세척한 다음, Red blood cell lysing buffer(Sigma, CA, USA)를 3분간 처리하여 적혈구를 제거 후 실험에 사용하였다.For splenocytes, a single cell suspension was prepared using tweezers and a mesh after the spleen of Wistar Rat was extracted. The single cell suspension was centrifuged three times (1,000g, 5 min, 4℃) with RPMI-1640 culture medium, washed, and then treated with Red blood cell lysing buffer (Sigma, CA, USA) for 3 minutes to remove red blood cells. was used for
또한 Natural Killer(NK) 세포인 AR42J 세포는 한국세포주은행(KCLB)에서 분양받아 사용하였다. 세포 배양에 필요한 시약은 Gibco(USA)에서 구입하였으며, 배지는 Roswell Park Memorial Institute 1640(RPMI-1640) 배지에 10% fetal bovine serum(FBS)과 1% antibiotics-antimycotic 제제를 첨가하여 5% CO2가 존재하는 37℃ 세포 배양기에서 2~3일에 한 번씩 계대 배양하면서 실험에 사용하였다.In addition, AR42J cells, which are Natural Killer (NK) cells, were purchased from the Korea Cell Line Bank (KCLB) and used. Reagents required for cell culture were purchased from Gibco (USA), and the medium was 5% CO 2 by adding 10% fetal bovine serum (FBS) and 1% antibiotics-antimycotic to Roswell Park Memorial Institute 1640 (RPMI-1640) medium. It was used for the experiment while subcultured once every 2-3 days in a 37°C cell incubator in the presence of
1-2. 세포생존율 분석1-2. Cell viability analysis
분리된 비장세포는 5×105 cells/90μl/well 농도로, AR42J 세포는 5×105 cells/90μl/well 농도로 농도별 추출물과 함께 처리하여 37℃, 5% CO2 농도에서 각각 24시간 동안 배양하였다. 이 후, 세포 배양액 100μl에 WST-1 시료 용액 10μl를 첨가하고 1시간 동안 배양하여 Multi Detection Reader(Infinite 200, TECAN Group Ltd, Switzerland)를 이용하여 흡광도 값을 측정하였다. 대조군은 시료를 처리하지 않고 시료를 용해한 용매만을 고농도 시험군과 동일 농도로 처리한 시험군으로 설정하였다. 세포의 증식율은 다음의 식에 따라 계산하였다.The separated spleen cells 5 × 10 5 cells / in 90μl / well density, AR42J cells are 5 × 10 5 cells / in a 90μl / well concentration treatment with concentrations extract respectively for 24 hours at 37 ℃, 5% CO 2 concentration incubated during Thereafter, 10 μl of the WST-1 sample solution was added to 100 μl of the cell culture medium and incubated for 1 hour to measure the absorbance value using a Multi Detection Reader (
Cell proliferation assay(%)=(시료 처리군의 흡광도/대조군의 흡광도)×100Cell proliferation assay (%) = (absorbance of sample treatment group / absorbance of control group) × 100
1-3. Cy 처리에 의한 비장세포 증식률 분석1-3. Analysis of Splenocyte Proliferation Rate by Cy Treatment
Cy(cyclophosphamide) 화합물의 처리에 따른 세포 독성과 이에 따른 비장세포의 생존률 감소 현상에 대하여 본 발명의 추출물 처리를 통한 비장세포의 생존율 증가 현상을 측정하였다.For the cytotoxicity according to the treatment of the Cy (cyclophosphamide) compound and the decrease in the viability of the splenocytes, the increase in the viability of the splenocytes through the treatment of the extract of the present invention was measured.
이와 같은 면역증진 효과를 관찰하기 위하여 분리한 비장세포를 96 well plate에 2×105 cells/90μl/well로 분주하고 24시간 배양하였다. 이 후, 대조군으로 Cy 화합물 1.6 mg/ml 농도를 처리하여 37℃, 5% CO2 조건에서 48시간 동안 배양하였고, 실험군으로 쌍별귀뚜라미(추출물, 초음파 추출물)와 갈색거저리(추출물, 초음파 추출물)를 100:0, 75:25, 50:50, 25:75, 및 0:100의 중량비로 처리하여 비장세포 증식률을 분석하였다.In order to observe this immune-enhancing effect, the isolated splenocytes were aliquoted at 2×10 5 cells/90 μl/well in a 96 well plate and cultured for 24 hours. Thereafter, as a control, a concentration of 1.6 mg/ml of Cy compound was treated and cultured for 48 hours at 37° C. and 5% CO 2 conditions. As an experimental group, twin-star cricket (extract, ultrasonic extract) and brown mealworm (extract, ultrasonic extract) were used. Splenocyte proliferation rates were analyzed by treatment with weight ratios of 100:0, 75:25, 50:50, 25:75, and 0:100.
1-4. NK cell activity 분석1-4. NK cell activity analysis
Natural Killer(NK) 세포인 AR42J 세포는 96 well plate에 5ⅹ106 cells/ml로 쌍별귀뚜라미(추출물, 초음파 추출물)와 갈색거저리(추출물, 초음파 추출물)를 75:25의 중량비로 혼합한 시료를 농도별로 처리하였다. AR42J cells, which are Natural Killer (NK) cells, were prepared by mixing a sample of 5 ×10 6 cells/ml in a 96 well plate with twin-star cricket (extract, ultrasonic extract) and brown mealworm (extract, ultrasonic extract) in a weight ratio of 75:25. processed.
배양 후 LDH 측정은 CytoTox detection kit(Takara)를 사용하여 측정하였으며, 반응액 내에 NAD의 산화로 의해 형성된 포마잔(formazan)을 490nm에서 흡광도를 측정하여 대조군과 비교 측정하였다.After incubation, LDH was measured using a CytoTox detection kit (Takara), and absorbance of formazan formed by oxidation of NAD in the reaction solution was measured at 490 nm and compared with the control.
1-5. 사이토카인 함량분석1-5. Cytokine content analysis
Cy 처리에 의한 면역세포의 사이토카인 분비량은 R&D system(Minneapolis, MN, USA)에서 구입한 ELISA kit를 이용하여 Cy 화합물과 쌍별귀뚜라미(추출물, 초음파 추출물)와 갈색거저리(추출물, 초음파 추출물)를 100:0, 75:25, 50:50, 25:75, 및 0:100의 중량비로 혼합한 시료를 농도별로 처리하여 24시간 반응 후 TNF-α(tumor necrosis factor-α), 및 IL-2(interleukin-2) 사이토카인의 발현량 변화를 분석하였다. Cytokine secretion of immune cells by Cy treatment was measured using an ELISA kit purchased from R&D system (Minneapolis, MN, USA), and 100 of Cy compound, paired crickets (extract, ultrasonic extract) and brown mealworm (extract, ultrasonic extract) were used. : 0, 75:25, 50:50, 25:75, and after 24 hours reaction by treating samples mixed at a weight ratio of 0:100 by concentration, TNF-α (tumor necrosis factor-α), and IL-2 ( Interleukin-2) cytokine expression level was analyzed.
1-6. CTL(Cytotoxic T Lymphocyte) 분석1-6. Cytotoxic T Lymphocyte (CTL) Analysis
비장 세포 부유액을 RPMI-1640(10% FBS, 1% penicillin-streptomycin) 배지로 희석하고, lysing buffer(BD PharmLyseTM Lysis Buffer, BD Biosciences, USA)에 3분간 현탁 시켜 적혈구를 제거한 뒤 원심세척하였다. 이 후, 96 well plate에 RPMI-1640(10% FBS, 1% penicillin-streptomycin) 배지로 희석하여 분리된 세포(effector cell)를 접종하였다. 표적 세포로는 HL-60 세포(Target cell)를 이용하여 effector cell과 target cell을 첨가한 후 37℃, 5% CO2 incubator에서 24시간 배양하였다. 배양 후 면역세포 증식능을 알아보기 위해 WST-1 시료를 각 well에 10ul 처리하여 ELISA reader로 540nm에서 측정하였다.The spleen cell suspension was diluted with RPMI-1640 (10% FBS, 1% penicillin-streptomycin) medium, suspended in lysing buffer (BD PharmLyse TM Lysis Buffer, BD Biosciences, USA) for 3 minutes to remove red blood cells and then centrifuged. Thereafter, the 96-well plate was diluted with RPMI-1640 (10% FBS, 1% penicillin-streptomycin) medium and the separated cells (effector cells) were inoculated. As target cells, effector cells and target cells were added using HL-60 cells (Target cells), and then cultured at 37° C., 5% CO 2 in an incubator for 24 hours. After culturing, 10ul of WST-1 sample was treated in each well to determine the ability to proliferate immune cells, and was measured at 540nm with an ELISA reader.
1-7. 통계 분석1-7. statistical analysis
모든 실험결과는 통계 프로그램(SPSS ver.12.0, SPSS Inc., Chicago, IL, USA)을 이용하여 평균 ± 표준오차(Mean ± S.E.)로 계산하였다. 각 실험군 간의 통계적 유의성 검정에 따른 통계분석은 ANOVA(one-way analysis of variance test)를 실시한 후 유의성이 있는 경우, p<0.05 미만일 때 Ducan's multiple range test로 사후 검정하였다.All experimental results were calculated as mean ± standard error (Mean ± S.E.) using a statistical program (SPSS ver.12.0, SPSS Inc., Chicago, IL, USA). Statistical analysis according to the statistical significance test between each experimental group was carried out after ANOVA (one-way analysis of variance test), and when there was significance, when p<0.05 was less than p<0.05, a post-hoc test was performed with Ducan's multiple range test.
실시예 1. 쌍별귀뚜라미와 갈색거저리의 추출물 및 초음파 추출물의 제조Example 1. Preparation of extracts and ultrasonic extracts of twin-star cricket and brown mealworm
쌍별귀뚜라미 추출물 시료는 분말 5g에 35ml PBS를 혼합하여 4℃에서 24시간동안 침지하여 상층액 사용하였고, 갈색거저리(고소애) 추출물 시료는 분말 5g에 35ml PBS를 혼합하여 4℃에서 24시간 동안 침지하여 상등액을 사용하였다. A sample of the cricket squirrel extract was mixed with 5 g of powder and 35 ml of PBS and immersed at 4 ° C for 24 hours to use the supernatant, and for the brown mealworm extract sample, 35 ml of PBS was mixed with 5 g of powder and immersed at 4 ° C for 24 hours. Thus, the supernatant was used.
실험에 사용한 추출물 시료는 0.22 μm syringe filter로 여과하여 사용하였고, 건조 중량을 측정한 결과 쌍별귀뚜라미는 22mg/ml, 갈색거저리는 28.9mg/ml로 측정되었다. 각 시료의 농도별 희석은 PBS로 하였으며 제조된 시료는 냉장 보관하였다. The extract sample used in the experiment was filtered with a 0.22 μm syringe filter, and as a result of measuring the dry weight, 22mg/ml for twin-star cricket and 28.9mg/ml for brown mealworm. The concentration of each sample was diluted with PBS, and the prepared sample was refrigerated.
쌍별귀뚜라미 초음파 추출물 시료는 50g에 450ml PBS를 혼합하여 저온 초음파 추출하였으며, 고소애 초음파 추출물 시료 또한 50g에 450ml PBS를 혼합하여 90분 동안 저온 초음파 추출하여 사용하였다. 초음파 추출물 시료는 실험에 사용하기 전에 0.22 μm syringe filter로 여과하였으며, 건조 중량을 측정한 결과 쌍별 추출물은 26.4mg/ml, 고소애 추출물은 32.1mg/ml이 측정되었다. 각 시료의 농도별 희석은 PBS로 하였으며 제조된 시료는 냉장 보관하였다. Ultrasonic cricket extract samples were obtained by mixing 50 g with 450 ml of PBS and ultrasonically extracted at low temperature, and the ultrasonic extract samples from Goae were also mixed with 50 g of 450 ml PBS and subjected to low temperature ultrasonic extraction for 90 minutes. The ultrasonic extract sample was filtered with a 0.22 μm syringe filter before use in the experiment, and as a result of measuring the dry weight, 26.4 mg/ml of the pairwise extract and 32.1 mg/ml of the Soae extract were measured. The concentration of each sample was diluted with PBS, and the prepared sample was refrigerated.
실시예 2. 쌍별귀뚜라미 및 갈색거저리 혼합물의 비장세포 독성 확인Example 2. Confirmation of splenocyte toxicity of a mixture of crickets and brown mealworm
쌍별귀뚜라미 및 갈색거저리(고소애)의 추출물 처리에 따른 비장세포의 독성을 확인하기 위해 세포 생존율을 분석하였다. 시료 노출은 24시간과 48시간 후 대조군에 대한 쌍별귀뚜라미와 갈색거저리 추출물을 처리한 시험군의 세포 생존율을 1~1000ug/ml의 농도로 실험하였다. 쌍별귀뚜라미 추출물은 500ug/ml의 농도부터 비장세포에서 독성을 나타내었고, 갈색거저리는 1000ug/ml 농도에서부터 세포독성이 확인되었다(도 1 참조). 이러한 결과를 토대로 쌍별귀뚜라미와 갈색거저리의 추출물은 각각 300ug/ml, 및 500ug/ml 이하의 농도를 최고농도로 설정하여 이하 실험을 진행하였다.Cell viability was analyzed to determine the toxicity of splenocytes following the treatment of extracts from paired crickets and brown mealworm (Gosoae). The sample exposure was tested at a concentration of 1-1000ug/ml for the cell viability of the test group treated with the extracts of pairwise crickets and brown mealworm for the control group after 24 and 48 hours. The pairwise cricket extract showed toxicity in splenocytes at a concentration of 500 ug/ml, and cytotoxicity was confirmed in brown mealworm at a concentration of 1000 ug/ml (see FIG. 1). Based on these results, the following experiments were carried out by setting the concentrations of 300 ug/ml and 500 ug/ml or less as the highest concentrations for the extracts of twin-star cricket and brown mealworm, respectively.
또한, 쌍별귀뚜라미 및 갈색거저리(고소애)의 초음파 추출물 처리에 따른 비장세포의 독성을 확인하기 위해 세포 생존율을 분석하였다. 시료 노출은 24시간과 48시간 후 대조군에 대한 쌍별귀뚜라미와 갈색거저리 초음파 추출물을 처리한 시험군의 세포 생존율을 1~1000ug/ml의 농도로 실험하였다. 쌍별귀뚜라미 초음파 추출물은 100ug/ml의 농도부터 비장세포에서 독성을 나타내었고, 갈색거저리는 300ug/ml 농도에서부터 세포독성이 확인되었다(도 2 참조). 이러한 결과를 토대로 쌍별귀뚜라미와 갈색거저리의 초음파 추출물은 각각 50ug/ml, 및 100ug/ml 이하의 농도를 최고농도로 설정하여 이하 실험을 진행하였다.In addition, the cell viability was analyzed to confirm the toxicity of splenocytes following the ultrasonic extract treatment of paired crickets and brown mealworm (Kosoae). After 24 and 48 hours of sample exposure, the cell viability of the test group treated with the ultrasonic extracts of paired cricket and brown mealworm for the control group was tested at a concentration of 1-1000ug/ml. Ultrasonic cricket extract was toxic to spleen cells at a concentration of 100 ug/ml, and cytotoxicity was confirmed from a concentration of 300 ug/ml in brown mealworm (see FIG. 2). Based on these results, the ultrasonic extracts of twin-star cricket and brown mealworm were respectively set to 50 ug/ml and 100 ug/ml or less as the highest concentrations, and the following experiments were carried out.
그리고, 쌍별귀뚜라미와 갈색거저리의 혼합물(75:25 w/w) 처리에 따른 비장세포의 증식률을 확인하기 위해 세포 생존율을 측정하였다. 실험은 24시간 후 대조군에 대한 쌍별귀뚜라미 및 갈색거저리의 혼합물을 처리한 시험군의 세포 증식률을 10~1000ug/ml의 농도별로 수행하였다. 쌍별귀뚜라미와 갈색거저리의 혼합 추출물은 500ug/ml의 농도에서 최고의 생존율을 보였고, 쌍별귀뚜라미와 갈색거저리 혼합 초음파 추출물은 100ug/ml에서 최고의 생존율을 나타내었다(도 3 참조). In addition, cell viability was measured to confirm the proliferation rate of splenocytes following treatment with a mixture of paired crickets and brown mealworm (75:25 w/w). After 24 hours, the cell proliferation rate of the test group treated with a mixture of paired crickets and brown mealworm for the control group was performed at a concentration of 10-1000ug/ml. The mixed extract of cricket serrata and brown mealworm exhibited the highest survival rate at a concentration of 500 ug/ml, and the mixed ultrasonic extract of cricket locust and brown mealworm showed the highest survival rate at 100 ug/ml (see FIG. 3).
실시예 3. Cy 처리에 의한 비장세포 생존률 분석Example 3. Analysis of splenocyte viability by Cy treatment
Cyclophosphamide(Cy) 화합물 처리에 의한 독성을 경감하는데 쌍별귀뚜라미와 갈색거저리(고소애)의 비율별 영향을 알아보기 위해 비장세포에 쌍별귀뚜라미와 갈색거저리의 혼합 시료와 Cy를 함께 처리하여 세포 생존률을 분석하였다. In order to investigate the effect of each ratio of cricket crickets and mealworms in the cyclophosphamide (Cy) compound in reducing toxicity, cell viability was analyzed by treating splenocytes with a mixed sample of crickets crickets and worms and Cy. did.
분석 결과 Cy 화합물을 처리한 시험군은 대조군과 비교하여 24시간 경과 후 62.1±0.7%, 48시간 경과 시 33.6±0.5%로 비장세포의 증식률이 현저히 감소하는 것으로 나타났다. 반면, Cy 화합물과 쌍별귀뚜라미와 갈색거저리(고소애)의 혼합 추출물과 혼합 혼합 추출물의 비율별 처리군은 24시간과 48시간 노출 시, 비율에 따라 세포 증식률 증가에서 차이를 보이나 모든 농도에서 유의적으로 증가되는 것을 확인 할 수 있었다(도 4 참조).As a result of the analysis, the test group treated with the Cy compound showed a significant decrease in the proliferation rate of splenocytes to 62.1±0.7% after 24 hours and 33.6±0.5% after 48 hours, compared to the control group. On the other hand, the ratio-treated group of the Cy compound and the mixed extract of crickets and brown mealworm (Gosoae) showed a difference in the increase in cell proliferation rate according to the ratio when exposed for 24 hours and 48 hours, but was significant at all concentrations. was confirmed to be increased (see FIG. 4).
상기 결과를 종합해 보면, 쌍별귀뚜라미와 갈색거저리(고소애)의 혼합 추출물과, 혼합 초음파 추출물의 비율별 cyclophosphamide(Cy) 화합물에 의한 독성 경감 효과는 쌍별귀뚜라미와 갈색거저리를 75:25의 중량비로 24시간 처리한 군에서 가장 좋은 효과를 보이는 것으로 확인됨에 따라 이후 실험은 75:25 중량비와 24시간 처리 조건에서 추가 효능 평가를 수행하였다.Summarizing the above results, the toxicity reduction effect of the cyclophosphamide (Cy) compound for each ratio of the mixed extract of cricket cricket and brown mealworm (Ko Soae) and the mixed ultrasonic extract was found to be 75:25 in weight ratio of twin cricket and brown mealworm. As it was confirmed that the group treated for 24 hours showed the best effect, further efficacy evaluation was performed in the following experiment at a 75:25 weight ratio and 24 hours treatment condition.
상기 결과를 토대로 쌍별귀뚜라미와 갈색거저리의 혼합 추출물과 혼합 초음파 추출물의 혼합 비율을 75:25 중량비로 고정하고, cyclophosphamide(Cy) 화합물을 처리한 비장세포에서 농도별 혼합 추출물과 혼합 초음파 추출물의 처리에 따른 비장세포의 생존율을 분석하였다. Based on the above results, the mixing ratio of the mixed extract and the mixed ultrasonic extract of paired cricket and brown mealworm was fixed at a weight ratio of 75:25, and in the splenocytes treated with the cyclophosphamide (Cy) compound, the concentration of the mixed extract and the mixed ultrasonic extract were applied. The viability of splenocytes was analyzed accordingly.
분석 결과 Cy 화합물을 처리한 시험군은 대조군과 비교하여 24시간 경과 시 55.1±0.6%로 비장세포의 증식률이 현저히 감소되었다. 반면, Cy 화합물과 혼합 추출물 및 혼합 초음파 추출물의 농도별 처리군은 농도 의존적으로 세포 증식률도 점차 증가하는 것으로 확인되었다(도 5 참조). As a result of the analysis, the test group treated with the Cy compound significantly reduced the proliferation rate of splenocytes to 55.1±0.6% after 24 hours compared to the control group. On the other hand, in the concentration-dependent treatment group of the Cy compound and the mixed extract and the mixed ultrasonic extract, it was confirmed that the cell proliferation rate also gradually increased (see FIG. 5 ).
이러한 결과를 통해, 본 발명의 쌍별귀뚜라미와 갈색거저리의 혼합물이 Cy 화합물의 처리로 저하된 면역세포의 활성을 실질적으로 증가시켜주고, 이에 따라 전반적으로 면역력을 개선하는 효과를 나타내는 것을 확인하여 준다. Through these results, it is confirmed that the mixture of crickets and brown mealworm of the present invention substantially increases the activity of immune cells decreased by the treatment of the Cy compound, and thus exhibits the effect of improving the overall immunity.
실시예 4. Cy 처리에 의한 사이토카인(TNF-α, IL-2) 발현량 분석Example 4. Cytokine (TNF-α, IL-2) expression level analysis by Cy treatment
Cyclophosphamide(Cy) 화합물 처리로 감소한 사이토카인 TNF-α, 및 IL-2 발현량 감소에 쌍별귀뚜라미와 갈색거저리(고소애) 추출물 및 초음파 추출물의 비율별 처리에 따른 발현 변화를 측정하였다. Cyclophosphamide (Cy) compound treatment reduced the expression level of cytokines TNF-α, and IL-2 decreased, and the expression change according to the ratio of cricket crickets and brown mealworm (Gosoae) extracts and ultrasonic extracts was measured.
비장세포에 쌍별귀뚜라미와 갈색거저리(고소애) 추출물 및 초음파 추출물 시료와 Cy 화합물을 함께 처리하고, 24시간 후에 비장세포의 사이토카인(TNF-α, IL-2) 발현량 변화를 ELISA 키트를 사용하여 분석하였다. Splenocytes were treated with spleen cricket and brown mealworm extract, ultrasonic extract, and Cy compound, and after 24 hours, the change in the expression level of splenocytes of cytokines (TNF-α, IL-2) was measured using an ELISA kit. and analyzed.
분석 결과, Cy 화합물을 처리한 시험군은 대조군과 비교하여 24시간 경과 시 비장세포의 사이토카인(TNF-α, IL-2) 발현량이 현저히 감소하는 것으로 나타났다. 반면 Cy 화합물과 쌍별귀뚜라미와 갈색거저리(고소애) 추출물의 비율별 처리군은 농도 의존적으로 비장세포에서 사이토카인(TNF-α, IL-2)의 발현량이 증가하는 것으로 확인되었다(도 6 참조). As a result of the analysis, it was found that the test group treated with the Cy compound significantly decreased the expression level of splenocytes (TNF-α, IL-2) for 24 hours compared to the control group. On the other hand, it was confirmed that the expression levels of cytokines (TNF-α, IL-2) in splenocytes increased in a concentration-dependent manner in the group treated with the Cy compound and the extracts of cricket and brown mealworm (Kosoae) in a concentration-dependent manner (see FIG. 6). .
또한, Cy 화합물과 쌍별귀뚜라미와 갈색거저리(고소애) 추출물의 비율별 처리군에서도 농도 의존적으로 비장세포에서 사이토카인(TNF-α, IL-2)의 발현량이 증가하는 것으로 확인되었다(도 7 참조).In addition, it was confirmed that the expression level of cytokines (TNF-α, IL-2) in splenocytes increased in a concentration-dependent manner in the treatment group by ratio of the Cy compound and extracts of cricket and brown mealworms (see Fig. 7). ).
상기와 같은 결과를 토대로 쌍별귀뚜라미와 갈색거저리의 추출물과 초음파 추출물의 cyclophosphamide(Cy) 화합물에 처리에 의한 TNF-α, 및 IL-2 사이토카인의 발현은 쌍별귀뚜라미와 갈색거저리가 75:25의 중량비로 혼합한 추출물과 같은 중량 비율의 초음파 추출물을 24시간 처리한 군에서 가장 좋은 효과를 보이는 것으로 확인함에 따라 이후 실험은 75:25 비율과 24시간으로 농도별 실험과 추가 효능 평가를 수행하였다.Based on the above results, the expression of TNF-α and IL-2 cytokines by treatment with the cyclophosphamide (Cy) compound of the extracts and ultrasonic extracts of crickets crickets and brown mealworms was 75:25 in weight ratio of crickets and brown mealworms. As it was confirmed that the best effect was shown in the group treated with the ultrasonic extract in the same weight ratio as the extract mixed with , for 24 hours, subsequent experiments were conducted at the 75:25 ratio and 24 hours, by concentration and additional efficacy evaluation.
상기 결과를 토대로 쌍별귀뚜라미와 갈색거저리의 혼합 추출물과 혼합 초음파 추출물의 혼합 비율을 75:25 중량비로 고정하고, cyclophosphamide(Cy) 화합물을 처리한 비장세포에서 농도별 혼합물의 처리에 따른 TNF-α 및 IL-2 사이토카인의 발현량 변화를 분석하였다. Based on the above results, the mixing ratio of the mixed extract and the mixed ultrasonic extract of paired cricket and brown mealworm was fixed at a weight ratio of 75:25, and in splenocytes treated with cyclophosphamide (Cy) compound, TNF-α and Changes in the expression level of IL-2 cytokines were analyzed.
분석 결과, Cy 화합물을 처리한 시험군은 대조군과 비교하여 24시간 경과 시 비장세포에서 사이토카인(TNF-α, IL-2) 발현량이 현저히 감소하는 것으로 나타났다. As a result of the analysis, it was found that the test group treated with the Cy compound significantly reduced the expression level of cytokines (TNF-α, IL-2) in splenocytes after 24 hours compared to the control group.
반면, Cy 화합물과 쌍별귀뚜라미 및 갈색거저리 혼합 추출물(도 8 참조) 및 혼합 초음파 추출물(도 9 참조)의 농도별 처리군은 농도 의존적으로 비장세포의 사이토카인(TNF-α, IL-2) 발현량이 점차 증가하는 것으로 확인되었다. On the other hand, the concentration-dependent treatment group of the Cy compound and the mixed extract (see FIG. 8) and the mixed ultrasonic extract (see FIG. 9) of paired cricket and brown mealworm (see FIG. 9) expressed splenocyte cytokine (TNF-α, IL-2) in a concentration-dependent manner. It was found that the amount gradually increased.
이러한 결과를 통해, 본 발명의 쌍별귀뚜라미와 갈색거저리의 혼합물이 Cy 화합물의 처리로 저하된 사이토카인(TNF-α, IL-2)의 발현을 실질적으로 증가시켜주고, 이에 따라 전반적으로 면역력을 개선하는 효과를 나타내는 것을 확인할 수 있었다. Through these results, the mixture of cricket and brown mealworm of the present invention substantially increases the expression of cytokines (TNF-α, IL-2) decreased by the treatment of the Cy compound, thus improving overall immunity It was confirmed that the effect of
실시예 5. NK(Natural Killer) 세포 활성 분석Example 5. NK (Natural Killer) Cell Activity Assay
쌍별귀뚜라미 및 갈색거저리(고소애)의 추출물 처리에 따른 Natural Killer(NK) 세포의 독성을 확인하기 위해 AR42J 세포의 생존율을 분석하였다. 시료 노출은 24시간 후 대조군에 대한 쌍별귀뚜라미와 갈색거저리 추출물을 처리한 시험군의 세포 생존율을 1~1000ug/ml의 농도로 실험하였다. The viability of AR42J cells was analyzed to determine the toxicity of Natural Killer (NK) cells following the treatment of extracts from paired crickets and brown mealworm (Gosoae). The sample exposure was tested at a concentration of 1-1000ug/ml for cell viability of the test group treated with the extracts of paired crickets and brown mealworm for the control group after 24 hours.
그 결과, 쌍별귀뚜라미 추출물은 500ug/ml의 농도부터 AR42J 세포에서 독성을 나타내었고, 갈색거저리 추출물은 1000ug/ml 농도에서부터 세포 독성이 확인되었다(도 10 참조). 이러한 결과를 토대로 NK 세포 활성을 측정하기 위하여 쌍별귀뚜라미와 갈색거저리의 추출물은 각각 300ug/ml, 및 500ug/ml 이하의 농도를 최고농도로 설정하여 이하 실험을 진행하였다.As a result, the cricket extract was cytotoxic from the concentration of 500 ug/ml to AR42J cells, and the Mealworm Mealworm extract showed cytotoxicity from the concentration of 1000 ug/ml (see FIG. 10 ). Based on these results, in order to measure the NK cell activity, the following experiments were conducted by setting the maximum concentrations of 300 ug/ml and 500 ug/ml or less of the extracts of paired cricket and brown mealworm, respectively.
또한, 쌍별귀뚜라미 및 갈색거저리(고소애)의 초음파 추출물 처리에 따른 AR42J 세포의 독성을 확인하기 위해 세포 생존율을 분석하였다. 시료 노출은 24시간과 48시간 후 대조군에 대한 쌍별귀뚜라미와 갈색거저리 초음파 추출물을 처리한 시험군의 세포 생존율을 1~1000ug/ml의 농도로 실험하였다. 쌍별귀뚜라미 초음파 추출물은 300ug/ml의 농도부터 AR42J 세포에서 독성을 나타내었고, 갈색거저리는 500ug/ml 농도에서부터 세포독성이 확인되었다(도 11 참조). 이러한 결과를 토대로 쌍별귀뚜라미와 갈색거저리의 초음파 추출물은 각각 100ug/ml, 및 300ug/ml 이하의 농도를 최고농도로 설정하여 이하 실험을 진행하였다.In addition, cell viability was analyzed in order to confirm the toxicity of AR42J cells following the ultrasonic extract treatment of paired crickets and brown mealworm (Gosoae). After 24 and 48 hours of sample exposure, the cell viability of the test group treated with the ultrasonic extracts of paired cricket and brown mealworm for the control group was tested at a concentration of 1-1000ug/ml. Ultrasonic extracts of paired crickets showed toxicity in AR42J cells at a concentration of 300 ug/ml, and cytotoxicity was confirmed in brown mealworms at a concentration of 500 ug/ml (see FIG. 11). Based on these results, the ultrasonic extracts of paired crickets and brown mealworms were respectively set at concentrations of 100 ug/ml and 300 ug/ml or less as the highest concentrations, and the following experiments were carried out.
그리고, 쌍별귀뚜라미와 갈색거저리의 혼합 비율에 따른 비장세포의 증식률을 확인하기 위해 혼합 비율(중량비 100:0, 75:25, 50:50; 25:75, 및 0:100)을 달리하여 AR42J 세포의 생존율을 측정하였다. And, in order to check the proliferation rate of splenocytes according to the mixing ratio of pairwise cricket and brown mealworm, AR42J cells were changed by varying the mixing ratio (weight ratio 100:0, 75:25, 50:50; 25:75, and 0:100). The survival rate was measured.
분석 결과, 쌍별귀뚜라미와 갈색거저리의 혼합 추출물과 혼합 초음파 추출물의 비율별 처리군은 대조군과 비교하여 24시간 동안 비율별로 세포독성에서 조금씩 차이를 보이나 모든 혼합 비율에서 유의적으로 증가하는 것을 확인할 수 있었다(도 12 참조). 실험 결과를 종합해 보면 쌍별귀뚜라미와 갈색거저리의 혼합 추출물과 혼합 초음파 추출물의 비율별 NK cell 활성은 이들을 75:25의 중량비로 혼합한 혼합물을 24시간 처리한 군에서 가장 좋은 효과를 보이는 것으로 확인함에 따라 이후 실험은 75:25 비율과 24시간으로 농도별 NK 세포 활성을 측정하였다.As a result of the analysis, it was confirmed that the group treated by ratio of the mixed extracts of cricket crickets and brown mealworm and the mixed ultrasonic extract showed a slight difference in cytotoxicity by ratio for 24 hours compared to the control group, but significantly increased in all mixing ratios. (See Fig. 12). Combining the experimental results, it was confirmed that the NK cell activity of the mixed extract and the mixed ultrasonic extract of twin-star cricket and brown mealworm showed the best effect in the group treated with the mixture mixed at a weight ratio of 75:25 for 24 hours. In subsequent experiments, NK cell activity was measured by concentration at a 75:25 ratio and 24 hours.
상기 결과를 토대로 쌍별귀뚜라미와 갈색거저리의 혼합 추출물과 혼합 초음파 추출물의 혼합 비율을 75:25 중량비로 고정하고, AR42J 세포의 생존율을 분석하였다. Based on the above results, the mixing ratio of the mixed extract of paired cricket and brown mealworm and the mixed ultrasonic extract was fixed at a weight ratio of 75:25, and the survival rate of AR42J cells was analyzed.
분석 결과, 쌍별귀뚜라미와 갈색거저리의 혼합물을 75:25의 중량비로 혼합한 혼합물을 처리한 군에서는 농도 의존적으로 AR42J 세포의 증식률이 유의적으로 증가하는 것을 확인 할 수 있었다(도 13 참조).As a result of the analysis, it was confirmed that the proliferation rate of AR42J cells was significantly increased in a concentration-dependent manner in the group treated with a mixture of a mixture of paired crickets and brown mealworm in a weight ratio of 75:25 (see FIG. 13).
이러한 결과를 통해, 본 발명의 쌍별귀뚜라미와 갈색거저리의 혼합물이 저하된 면역세포의 능력을 회복시켜줌에 따라 NK 세포의 활성을 증가시켜주는 것을 확인함으로써 본 발명의 쌍별귀뚜라미와 갈색거저리의 혼합물이 면역력 증가에 직접적인 효과를 나타내는 것을 확인할 수 있었다. Through these results, it was confirmed that the mixture of cricket and brown mealworm of the present invention increases the activity of NK cells as it restores the decreased ability of immune cells. It was confirmed that it had a direct effect on the increase.
실시예 6. CTL(Cytotoxic T Lymphocyte) 세포 활성 분석Example 6. Cytotoxic T Lymphocyte (CTL) Cell Activity Assay
쌍별귀뚜라미와 갈색거저리의 혼합 추출물과 혼합 초음파 추출물의 혼합 비율을 75:25 중량비로 고정하고, 혼합물이 CTL(Cytotoxic T Lymphocyte) 세포인 HL-60 세포의 생존에 미치는 영향을 분석하였다. The mixing ratio of the mixed extract of cricket cricket and brown mealworm and the mixed ultrasonic extract was fixed at a weight ratio of 75:25, and the effect of the mixture on the survival of CTL (Cytotoxic T Lymphocyte) cells, HL-60 cells, was analyzed.
시료 노출은 24시간 후 대조군에 대한 쌍별귀뚜라미와 갈색거저리 혼합 추출물을 처리한 시험군의 세포 생존율을 1~1000ug/ml의 농도로 실험하였다. After 24 hours of sample exposure, the cell viability of the test group treated with the mixed extract of paired cricket and brown mealworm for the control group was tested at a concentration of 1 to 1000 ug/ml.
그 결과, 쌍별귀뚜라미와 갈색거저리의 혼합 추출물(A)은 1,000ug/ml의 농도부터 세포독성을 보였고, 쌍별귀뚜라미와 갈색거저리의 혼합 초음파 추출물(B)은 500ug/ml에서 세포독성이 확인되었다(도 14 참조). 이러한 결과 토대로 쌍별귀뚜라미와 갈색거저리의 혼합 추출물과 혼합 초음파 추출물의 CTL 활성 측정은 각각 500ug/ml, 및 300ug/ml 이하의 농도를 최고농도로 설정하였다.As a result, the mixed extract (A) of twin-star cricket and brown mealworm showed cytotoxicity at a concentration of 1,000 ug/ml, and the mixed ultrasonic extract of twin-star cricket and brown mealworm (B) showed cytotoxicity at 500 ug/ml ( 14). Based on these results, the CTL activity measurement of the mixed extracts of cricket crickets and brown mealworm and the mixed ultrasonic extract was set to the highest concentrations of 500ug/ml and 300ug/ml or less, respectively.
또한, 쌍별귀뚜라미와 갈색거저리의 혼합 추출물과 혼합 초음파 추출물의 혼합 비율을 75:25 중량비로 고정하고, 비장세포에 혼합 시료들와 HL-60 세포를 함께 처리하여 세포 독성을 분석하였다. In addition, the mixing ratio of the mixed extract of cricket cricket and brown mealworm and the mixed ultrasonic extract was fixed at a weight ratio of 75:25, and cytotoxicity was analyzed by treating splenocytes with mixed samples and HL-60 cells.
분석 결과, 쌍별귀뚜라미와 갈색거저리의 혼합 추출물과 혼합 초음파 추출물의 농도별 처리군은 대조군과 비교하여 24시간 경과 시 농도 의존적으로 HL-60 세포의 세포 증식률을 유의적으로 증가시키는 것을 확인할 수 있었다(도 15 참조). As a result of the analysis, it was confirmed that the concentration-dependently increased cell proliferation rate of HL-60 cells was found to be significantly increased in the concentration-dependent treatment group of the mixed extract of cricket cricket and brown mealworm and the mixed ultrasonic extract for 24 hours compared to the control group ( 15).
이러한 결과는 쌍별귀뚜라미와 갈색거저리의 혼합물이 저하된 면역세포의 능력을 회복시키고, 이에 따라 CTL(Cytotoxic T Lymphocyte) 세포의 활성을 증가시켜, 세포 매개성 면역력 증가에 직접적으로 효과를 나타내는 것을 확인할 수 있었다. These results confirm that the mixture of paired crickets and brown mealworms restores the decreased ability of immune cells and, accordingly, increases the activity of CTL (Cytotoxic T Lymphocyte) cells, thereby showing a direct effect on the increase of cell-mediated immunity. there was.
이상으로 본 발명의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현예일 뿐이며, 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.As described above in detail a specific part of the present invention, for those of ordinary skill in the art, this specific description is only a preferred embodiment, and it is clear that the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.
Claims (5)
상기 쌍별귀뚜라미와 상기 갈색거저리는 70-80:20-30의 중량부로 혼합된 것을 특징으로 하는 면역증진용 식품 조성물.According to claim 1,
The food composition for enhancing immunity, characterized in that the pairwise cricket and the brown mealworm are mixed in a weight ratio of 70-80:20-30.
상기 조성물은 비장세포, NK(Natural Killer) 세포, 및 CTL(Cytotoxic T Lymphocyte) 세포의 세포 증식을 증진하는 것을 특징으로 하는 면역증진용 식품 조성물.According to claim 1,
The composition is a food composition for enhancing immunity, characterized in that it promotes cell proliferation of splenocytes, NK (Natural Killer) cells, and CTL (Cytotoxic T Lymphocyte) cells.
상기 조성물은 TNF-α 및 IL-2의 발현을 증가시키는 것을 특징으로 하는 면역증진용 식품 조성물.According to claim 1,
The composition is a food composition for enhancing immunity, characterized in that it increases the expression of TNF-α and IL-2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020190178881A KR20210085627A (en) | 2019-12-31 | 2019-12-31 | Composition for Enhancing Immunity Comprising Complex Extracts of Gryllus bimaculatus and Tenebrio molitor Linnaues as Active Ingredient |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020190178881A KR20210085627A (en) | 2019-12-31 | 2019-12-31 | Composition for Enhancing Immunity Comprising Complex Extracts of Gryllus bimaculatus and Tenebrio molitor Linnaues as Active Ingredient |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| KR20210085627A true KR20210085627A (en) | 2021-07-08 |
Family
ID=76894479
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020190178881A Ceased KR20210085627A (en) | 2019-12-31 | 2019-12-31 | Composition for Enhancing Immunity Comprising Complex Extracts of Gryllus bimaculatus and Tenebrio molitor Linnaues as Active Ingredient |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR20210085627A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20230102039A (en) | 2021-12-29 | 2023-07-07 | 단국대학교 천안캠퍼스 산학협력단 | Composition for enhancing immunity comprising gryllus bimaculatus prepared by enzymatic hydrolysis and manufacturing method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017213172A1 (en) | 2016-06-07 | 2017-12-14 | 国立大学法人愛媛大学 | Method for production of composition |
| JP2019054749A (en) | 2017-09-21 | 2019-04-11 | 株式会社愛南リベラシオ | Method of producing composition |
| KR20190125035A (en) | 2018-04-27 | 2019-11-06 | 가톨릭대학교 산학협력단 | Adjuvant comprising nucleic acid molecule inserted viral expression regulation sequence and phamraceutical composition having the adjuvant |
-
2019
- 2019-12-31 KR KR1020190178881A patent/KR20210085627A/en not_active Ceased
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017213172A1 (en) | 2016-06-07 | 2017-12-14 | 国立大学法人愛媛大学 | Method for production of composition |
| JP2019054749A (en) | 2017-09-21 | 2019-04-11 | 株式会社愛南リベラシオ | Method of producing composition |
| KR20190125035A (en) | 2018-04-27 | 2019-11-06 | 가톨릭대학교 산학협력단 | Adjuvant comprising nucleic acid molecule inserted viral expression regulation sequence and phamraceutical composition having the adjuvant |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20230102039A (en) | 2021-12-29 | 2023-07-07 | 단국대학교 천안캠퍼스 산학협력단 | Composition for enhancing immunity comprising gryllus bimaculatus prepared by enzymatic hydrolysis and manufacturing method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR20170019461A (en) | Composition with high content of cyclic dipeptide | |
| KR20210063728A (en) | Fermented kiwi powder for improving bowel function and method of preparing the same | |
| KR101870960B1 (en) | Composition for preventing or treating of colitis disease comprising Lactobacillus sakei K040706 as an active ingredient | |
| JP6443804B2 (en) | Natural killer cell activity promoter | |
| JP6086953B2 (en) | AMPK activator | |
| KR20120051707A (en) | Crude caffeine complex, improved food products using the crude caffeine complex, and methods of use thereof | |
| JPWO2004100969A1 (en) | Therapeutic agent | |
| JPWO2004112817A1 (en) | Celery family-derived extract and method for producing the same | |
| JP2017193497A (en) | Muscle enhancer | |
| KR102405386B1 (en) | Composition for Enhancing Immunity Comprising Complex Extracts of Swallow''s Nest as Active Ingredient | |
| KR102203626B1 (en) | Pharmaceutical and functional food composition comprising phosvitin and lysozyme for enhancing immunity | |
| KR101704123B1 (en) | Method for seperating bee venom containing active amines and food composition thereof | |
| KR20210085627A (en) | Composition for Enhancing Immunity Comprising Complex Extracts of Gryllus bimaculatus and Tenebrio molitor Linnaues as Active Ingredient | |
| KR102315959B1 (en) | A composition for reinforcing immune function comprising the extract of oat sprout | |
| KR20220109668A (en) | Composition for oral administration comprising extract of oxya chinensis sinuosa | |
| JP2017031120A (en) | TNF-α AND IL-6 PRODUCTION INHIBITORS, AND MUSCLE INFLAMMATORY INHIBITORS USING THE SAME | |
| JP6105186B2 (en) | Pancreatic lipase inhibitor | |
| KR101895158B1 (en) | A composition comprising mixture of the extract of Eucommia ulmoides Oliver and silkworm powder for improving immunity | |
| KR20170037690A (en) | Composition for enhancing blood circulation containing the extract of Rice Bran and Fermented Rice Bran as an active ingredient | |
| JP6954960B2 (en) | TNF-α and IL-6 production inhibitors and muscle inflammation inhibitors using them | |
| KR102244732B1 (en) | Probiotic acetic acid bacteria Acetobacter pasteurianus MGLV and its immunomodulatory effect | |
| JP7206623B2 (en) | Composition for prevention and improvement of glucose metabolism disorder | |
| KR20220128198A (en) | Immune enhancing composition comprising a mixture of Sanghwang mushroom extract and ginseng extract, and method for preparing the same | |
| KR101923822B1 (en) | Anti-inflammatory composition containing extract of cornus officinalis seed | |
| KR101832357B1 (en) | A composition for the enhancement of immune system comprising extracts of soybean sprouts as an active ingredient and the method of preparation thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PA0109 | Patent application |
Patent event code: PA01091R01D Comment text: Patent Application Patent event date: 20191231 |
|
| PA0201 | Request for examination | ||
| PG1501 | Laying open of application | ||
| E902 | Notification of reason for refusal | ||
| PE0902 | Notice of grounds for rejection |
Comment text: Notification of reason for refusal Patent event date: 20211021 Patent event code: PE09021S01D |
|
| E601 | Decision to refuse application | ||
| PE0601 | Decision on rejection of patent |
Patent event date: 20220119 Comment text: Decision to Refuse Application Patent event code: PE06012S01D Patent event date: 20211021 Comment text: Notification of reason for refusal Patent event code: PE06011S01I |