KR102604839B1 - Composition for antigenicity comprising peptide from PRRSV - Google Patents
Composition for antigenicity comprising peptide from PRRSV Download PDFInfo
- Publication number
- KR102604839B1 KR102604839B1 KR1020210046000A KR20210046000A KR102604839B1 KR 102604839 B1 KR102604839 B1 KR 102604839B1 KR 1020210046000 A KR1020210046000 A KR 1020210046000A KR 20210046000 A KR20210046000 A KR 20210046000A KR 102604839 B1 KR102604839 B1 KR 102604839B1
- Authority
- KR
- South Korea
- Prior art keywords
- peptide
- present
- seq
- virus
- respiratory syndrome
- Prior art date
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 110
- 241001135989 Porcine reproductive and respiratory syndrome virus Species 0.000 title claims abstract description 44
- 239000000203 mixture Substances 0.000 title claims abstract description 43
- 230000002163 immunogen Effects 0.000 claims abstract description 19
- 210000001744 T-lymphocyte Anatomy 0.000 claims abstract description 12
- 108010074328 Interferon-gamma Proteins 0.000 claims abstract description 11
- 229960003130 interferon gamma Drugs 0.000 claims abstract description 4
- 102000008070 Interferon-gamma Human genes 0.000 claims abstract 2
- 208000005342 Porcine Reproductive and Respiratory Syndrome Diseases 0.000 claims description 24
- 239000008194 pharmaceutical composition Substances 0.000 claims description 24
- 229960005486 vaccine Drugs 0.000 claims description 24
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 20
- 238000000034 method Methods 0.000 claims description 16
- 241000700605 Viruses Species 0.000 abstract description 30
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 abstract description 15
- 210000004027 cell Anatomy 0.000 abstract description 8
- 230000001939 inductive effect Effects 0.000 abstract description 8
- 230000007969 cellular immunity Effects 0.000 abstract description 6
- 230000003321 amplification Effects 0.000 abstract description 2
- 238000003199 nucleic acid amplification method Methods 0.000 abstract description 2
- 230000004936 stimulating effect Effects 0.000 abstract 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 23
- 241000282887 Suidae Species 0.000 description 11
- 208000015181 infectious disease Diseases 0.000 description 10
- 230000005727 virus proliferation Effects 0.000 description 10
- 102100037850 Interferon gamma Human genes 0.000 description 9
- 150000001413 amino acids Chemical class 0.000 description 9
- 239000000427 antigen Substances 0.000 description 9
- 102000036639 antigens Human genes 0.000 description 9
- 108091007433 antigens Proteins 0.000 description 9
- -1 olive oil Chemical compound 0.000 description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 7
- 230000005764 inhibitory process Effects 0.000 description 7
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 6
- 235000001014 amino acid Nutrition 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- 230000002401 inhibitory effect Effects 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 239000002671 adjuvant Substances 0.000 description 5
- 239000000969 carrier Substances 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 239000000839 emulsion Substances 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- 230000000638 stimulation Effects 0.000 description 5
- 239000000725 suspension Substances 0.000 description 5
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 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 4
- 239000000654 additive Substances 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 239000008121 dextrose Substances 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 230000028993 immune response Effects 0.000 description 4
- 239000013642 negative control Substances 0.000 description 4
- 230000002265 prevention Effects 0.000 description 4
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 239000002202 Polyethylene glycol Substances 0.000 description 3
- 239000003963 antioxidant agent Substances 0.000 description 3
- 235000006708 antioxidants Nutrition 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 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 3
- 230000005540 biological transmission Effects 0.000 description 3
- 239000007844 bleaching agent Substances 0.000 description 3
- 239000003086 colorant Substances 0.000 description 3
- 230000005847 immunogenicity Effects 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000003921 oil Substances 0.000 description 3
- 235000019198 oils Nutrition 0.000 description 3
- 238000007911 parenteral administration Methods 0.000 description 3
- 229920001223 polyethylene glycol Polymers 0.000 description 3
- 239000003755 preservative agent Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 230000028327 secretion Effects 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 229940124597 therapeutic agent Drugs 0.000 description 3
- GHOKWGTUZJEAQD-ZETCQYMHSA-N (D)-(+)-Pantothenic acid Chemical compound OCC(C)(C)[C@@H](O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-ZETCQYMHSA-N 0.000 description 2
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 2
- NLZUEZXRPGMBCV-UHFFFAOYSA-N Butylhydroxytoluene Chemical compound CC1=CC(C(C)(C)C)=C(O)C(C(C)(C)C)=C1 NLZUEZXRPGMBCV-UHFFFAOYSA-N 0.000 description 2
- ZKQDCIXGCQPQNV-UHFFFAOYSA-N Calcium hypochlorite Chemical compound [Ca+2].Cl[O-].Cl[O-] ZKQDCIXGCQPQNV-UHFFFAOYSA-N 0.000 description 2
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 2
- 241000255925 Diptera Species 0.000 description 2
- LVGKNOAMLMIIKO-UHFFFAOYSA-N Elaidinsaeure-aethylester Natural products CCCCCCCCC=CCCCCCCCC(=O)OCC LVGKNOAMLMIIKO-UHFFFAOYSA-N 0.000 description 2
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- 229930195725 Mannitol Natural products 0.000 description 2
- 108010052285 Membrane Proteins Proteins 0.000 description 2
- 102000018697 Membrane Proteins Human genes 0.000 description 2
- 101710144128 Non-structural protein 2 Proteins 0.000 description 2
- 208000037581 Persistent Infection Diseases 0.000 description 2
- 102100022648 Reticulon-2 Human genes 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 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 2
- 229930006000 Sucrose Natural products 0.000 description 2
- XLOMVQKBTHCTTD-UHFFFAOYSA-N Zinc monoxide Chemical compound [Zn]=O XLOMVQKBTHCTTD-UHFFFAOYSA-N 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 239000000443 aerosol Substances 0.000 description 2
- 239000008365 aqueous carrier Substances 0.000 description 2
- 239000003125 aqueous solvent Substances 0.000 description 2
- 239000007900 aqueous suspension Substances 0.000 description 2
- 229940031567 attenuated vaccine Drugs 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000011230 binding agent Substances 0.000 description 2
- 229960002685 biotin Drugs 0.000 description 2
- 235000020958 biotin Nutrition 0.000 description 2
- 239000011616 biotin Substances 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 239000011651 chromium Substances 0.000 description 2
- 239000010949 copper Substances 0.000 description 2
- 239000006071 cream Substances 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000007884 disintegrant Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- LVGKNOAMLMIIKO-QXMHVHEDSA-N ethyl oleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC LVGKNOAMLMIIKO-QXMHVHEDSA-N 0.000 description 2
- 229940093471 ethyl oleate Drugs 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 235000003599 food sweetener Nutrition 0.000 description 2
- 230000009036 growth inhibition Effects 0.000 description 2
- 229940031551 inactivated vaccine Drugs 0.000 description 2
- 230000009349 indirect transmission Effects 0.000 description 2
- 230000002458 infectious effect Effects 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 229910052500 inorganic mineral Inorganic materials 0.000 description 2
- 238000007918 intramuscular administration Methods 0.000 description 2
- 238000001990 intravenous administration Methods 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000000314 lubricant Substances 0.000 description 2
- 239000011777 magnesium Substances 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 239000011572 manganese Substances 0.000 description 2
- 239000000594 mannitol Substances 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 239000011707 mineral Substances 0.000 description 2
- 210000004980 monocyte derived macrophage Anatomy 0.000 description 2
- LPUQAYUQRXPFSQ-DFWYDOINSA-M monosodium L-glutamate Chemical compound [Na+].[O-]C(=O)[C@@H](N)CCC(O)=O LPUQAYUQRXPFSQ-DFWYDOINSA-M 0.000 description 2
- 235000013923 monosodium glutamate Nutrition 0.000 description 2
- 210000003205 muscle Anatomy 0.000 description 2
- 239000002674 ointment Substances 0.000 description 2
- 239000004006 olive oil Substances 0.000 description 2
- 235000008390 olive oil Nutrition 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 210000002826 placenta Anatomy 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 2
- 239000012679 serum free medium Substances 0.000 description 2
- 235000010288 sodium nitrite Nutrition 0.000 description 2
- GEHJYWRUCIMESM-UHFFFAOYSA-L sodium sulfite Chemical compound [Na+].[Na+].[O-]S([O-])=O GEHJYWRUCIMESM-UHFFFAOYSA-L 0.000 description 2
- 239000000600 sorbitol Substances 0.000 description 2
- 235000010356 sorbitol Nutrition 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- 229960004793 sucrose Drugs 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 239000003765 sweetening agent Substances 0.000 description 2
- 239000006188 syrup Substances 0.000 description 2
- 235000020357 syrup Nutrition 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- 235000015112 vegetable and seed oil Nutrition 0.000 description 2
- 239000008158 vegetable oil Substances 0.000 description 2
- 230000005570 vertical transmission Effects 0.000 description 2
- 239000001993 wax Substances 0.000 description 2
- ASWBNKHCZGQVJV-UHFFFAOYSA-N (3-hexadecanoyloxy-2-hydroxypropyl) 2-(trimethylazaniumyl)ethyl phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(O)COP([O-])(=O)OCC[N+](C)(C)C ASWBNKHCZGQVJV-UHFFFAOYSA-N 0.000 description 1
- CHHHXKFHOYLYRE-UHFFFAOYSA-M 2,4-Hexadienoic acid, potassium salt (1:1), (2E,4E)- Chemical compound [K+].CC=CC=CC([O-])=O CHHHXKFHOYLYRE-UHFFFAOYSA-M 0.000 description 1
- UFBJCMHMOXMLKC-UHFFFAOYSA-N 2,4-dinitrophenol Chemical compound OC1=CC=C([N+]([O-])=O)C=C1[N+]([O-])=O UFBJCMHMOXMLKC-UHFFFAOYSA-N 0.000 description 1
- 206010000234 Abortion spontaneous Diseases 0.000 description 1
- CKLDHDOIYBVUNP-KBIXCLLPSA-N Ala-Ile-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(O)=O CKLDHDOIYBVUNP-KBIXCLLPSA-N 0.000 description 1
- YCRAFFCYWOUEOF-DLOVCJGASA-N Ala-Phe-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C)CC1=CC=CC=C1 YCRAFFCYWOUEOF-DLOVCJGASA-N 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- APKFDSVGJQXUKY-KKGHZKTASA-N Amphotericin-B Natural products O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1C=CC=CC=CC=CC=CC=CC=C[C@H](C)[C@@H](O)[C@@H](C)[C@H](C)OC(=O)C[C@H](O)C[C@H](O)CC[C@@H](O)[C@H](O)C[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 APKFDSVGJQXUKY-KKGHZKTASA-N 0.000 description 1
- 229920000945 Amylopectin Polymers 0.000 description 1
- HJAICMSAKODKRF-GUBZILKMSA-N Arg-Cys-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O HJAICMSAKODKRF-GUBZILKMSA-N 0.000 description 1
- HQIZDMIGUJOSNI-IUCAKERBSA-N Arg-Gly-Arg Chemical compound N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O HQIZDMIGUJOSNI-IUCAKERBSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 239000004215 Carbon black (E152) Substances 0.000 description 1
- GHOKWGTUZJEAQD-UHFFFAOYSA-N Chick antidermatitis factor Natural products OCC(C)(C)C(O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-UHFFFAOYSA-N 0.000 description 1
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 description 1
- 208000003322 Coinfection Diseases 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 235000019739 Dicalciumphosphate Nutrition 0.000 description 1
- 206010061818 Disease progression Diseases 0.000 description 1
- GGLIEWRLXDLBBF-UHFFFAOYSA-N Dulcin Chemical compound CCOC1=CC=C(NC(N)=O)C=C1 GGLIEWRLXDLBBF-UHFFFAOYSA-N 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 239000004278 EU approved seasoning Substances 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 239000004386 Erythritol Substances 0.000 description 1
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 208000001951 Fetal Death Diseases 0.000 description 1
- 239000004606 Fillers/Extenders Substances 0.000 description 1
- 206010055690 Foetal death Diseases 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- BPLNJYHNAJVLRT-ACZMJKKPSA-N Glu-Ser-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O BPLNJYHNAJVLRT-ACZMJKKPSA-N 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- ORZGPQXISSXQGW-IHRRRGAJSA-N His-His-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C(C)C)C(O)=O ORZGPQXISSXQGW-IHRRRGAJSA-N 0.000 description 1
- DGTOKVBDZXJHNZ-WZLNRYEVSA-N Ile-Thr-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N DGTOKVBDZXJHNZ-WZLNRYEVSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 208000029523 Interstitial Lung disease Diseases 0.000 description 1
- 244000017020 Ipomoea batatas Species 0.000 description 1
- 235000002678 Ipomoea batatas Nutrition 0.000 description 1
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 1
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- WSGXUIQTEZDVHJ-GARJFASQSA-N Leu-Ala-Pro Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@@H]1C(O)=O WSGXUIQTEZDVHJ-GARJFASQSA-N 0.000 description 1
- PPBKJAQJAUHZKX-SRVKXCTJSA-N Leu-Cys-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@H](C(O)=O)CC(C)C PPBKJAQJAUHZKX-SRVKXCTJSA-N 0.000 description 1
- BABSVXFGKFLIGW-UWVGGRQHSA-N Leu-Gly-Arg Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCNC(N)=N BABSVXFGKFLIGW-UWVGGRQHSA-N 0.000 description 1
- YOKVEHGYYQEQOP-QWRGUYRKSA-N Leu-Leu-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YOKVEHGYYQEQOP-QWRGUYRKSA-N 0.000 description 1
- HOMFINRJHIIZNJ-HOCLYGCPSA-N Leu-Trp-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)NCC(O)=O HOMFINRJHIIZNJ-HOCLYGCPSA-N 0.000 description 1
- IMDJSVBFQKDDEQ-MGHWNKPDSA-N Lys-Tyr-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](CCCCN)N IMDJSVBFQKDDEQ-MGHWNKPDSA-N 0.000 description 1
- GILLQRYAWOMHED-DCAQKATOSA-N Lys-Val-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCCN GILLQRYAWOMHED-DCAQKATOSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 239000005913 Maltodextrin Substances 0.000 description 1
- 229920002774 Maltodextrin Polymers 0.000 description 1
- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 description 1
- AFVOKRHYSSFPHC-STECZYCISA-N Met-Ile-Tyr Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 AFVOKRHYSSFPHC-STECZYCISA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229930193140 Neomycin Natural products 0.000 description 1
- 101100342977 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) leu-1 gene Proteins 0.000 description 1
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 1
- 101710144127 Non-structural protein 1 Proteins 0.000 description 1
- 206010048685 Oral infection Diseases 0.000 description 1
- 241000237988 Patellidae Species 0.000 description 1
- BYAIIACBWBOJCU-URLPEUOOSA-N Phe-Ile-Thr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O BYAIIACBWBOJCU-URLPEUOOSA-N 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 108010093965 Polymyxin B Proteins 0.000 description 1
- 102100031776 SH2 domain-containing protein 3A Human genes 0.000 description 1
- WINXNKPZLFISPD-UHFFFAOYSA-M Saccharin sodium Chemical compound [Na+].C1=CC=C2C(=O)[N-]S(=O)(=O)C2=C1 WINXNKPZLFISPD-UHFFFAOYSA-M 0.000 description 1
- 239000005708 Sodium hypochlorite Substances 0.000 description 1
- YGCDFAJJCRVQKU-RCWTZXSCSA-N Thr-Pro-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)[C@@H](C)O YGCDFAJJCRVQKU-RCWTZXSCSA-N 0.000 description 1
- MYNYCUXMIIWUNW-IEGACIPQSA-N Thr-Trp-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CCCCN)C(=O)O)N)O MYNYCUXMIIWUNW-IEGACIPQSA-N 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- IECQJCJNPJVUSB-IHRRRGAJSA-N Val-Tyr-Ser Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](Cc1ccc(O)cc1)C(=O)N[C@@H](CO)C(O)=O IECQJCJNPJVUSB-IHRRRGAJSA-N 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 206010058874 Viraemia Diseases 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 229940037003 alum Drugs 0.000 description 1
- WNROFYMDJYEPJX-UHFFFAOYSA-K aluminium hydroxide Chemical compound [OH-].[OH-].[OH-].[Al+3] WNROFYMDJYEPJX-UHFFFAOYSA-K 0.000 description 1
- 210000001132 alveolar macrophage Anatomy 0.000 description 1
- APKFDSVGJQXUKY-INPOYWNPSA-N amphotericin B Chemical compound O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1/C=C/C=C/C=C/C=C/C=C/C=C/C=C/[C@H](C)[C@@H](O)[C@@H](C)[C@H](C)OC(=O)C[C@H](O)C[C@H](O)CC[C@@H](O)[C@H](O)C[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 APKFDSVGJQXUKY-INPOYWNPSA-N 0.000 description 1
- 229960003942 amphotericin b Drugs 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000010775 animal oil Substances 0.000 description 1
- 239000002518 antifoaming agent Substances 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 239000004599 antimicrobial Substances 0.000 description 1
- 108010062796 arginyllysine Proteins 0.000 description 1
- 239000000022 bacteriostatic agent Substances 0.000 description 1
- 239000000440 bentonite Substances 0.000 description 1
- 229910000278 bentonite Inorganic materials 0.000 description 1
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000007975 buffered saline Substances 0.000 description 1
- CZBZUDVBLSSABA-UHFFFAOYSA-N butylated hydroxyanisole Chemical compound COC1=CC=C(O)C(C(C)(C)C)=C1.COC1=CC=C(O)C=C1C(C)(C)C CZBZUDVBLSSABA-UHFFFAOYSA-N 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- CJZGTCYPCWQAJB-UHFFFAOYSA-L calcium stearate Chemical compound [Ca+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O CJZGTCYPCWQAJB-UHFFFAOYSA-L 0.000 description 1
- 239000008116 calcium stearate Substances 0.000 description 1
- 235000013539 calcium stearate Nutrition 0.000 description 1
- BPKIGYQJPYCAOW-FFJTTWKXSA-I calcium;potassium;disodium;(2s)-2-hydroxypropanoate;dichloride;dihydroxide;hydrate Chemical compound O.[OH-].[OH-].[Na+].[Na+].[Cl-].[Cl-].[K+].[Ca+2].C[C@H](O)C([O-])=O BPKIGYQJPYCAOW-FFJTTWKXSA-I 0.000 description 1
- 229910002091 carbon monoxide Inorganic materials 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 229910052804 chromium Inorganic materials 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 235000008504 concentrate Nutrition 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- 239000000645 desinfectant Substances 0.000 description 1
- NEFBYIFKOOEVPA-UHFFFAOYSA-K dicalcium phosphate Chemical compound [Ca+2].[Ca+2].[O-]P([O-])([O-])=O NEFBYIFKOOEVPA-UHFFFAOYSA-K 0.000 description 1
- 229940038472 dicalcium phosphate Drugs 0.000 description 1
- 229910000390 dicalcium phosphate Inorganic materials 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000013681 dietary sucrose Nutrition 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 230000005750 disease progression Effects 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 239000008157 edible vegetable oil Substances 0.000 description 1
- 230000002500 effect on skin Effects 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 1
- 235000019414 erythritol Nutrition 0.000 description 1
- 229940009714 erythritol Drugs 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 229960004756 ethanol Drugs 0.000 description 1
- MVPICKVDHDWCJQ-UHFFFAOYSA-N ethyl 3-pyrrolidin-1-ylpropanoate Chemical compound CCOC(=O)CCN1CCCC1 MVPICKVDHDWCJQ-UHFFFAOYSA-N 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 239000010685 fatty oil Substances 0.000 description 1
- 231100000479 fetal death Toxicity 0.000 description 1
- 210000003754 fetus Anatomy 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 229940014144 folate Drugs 0.000 description 1
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000011724 folic acid Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000010855 food raising agent Nutrition 0.000 description 1
- 235000011194 food seasoning agent Nutrition 0.000 description 1
- 229960004279 formaldehyde Drugs 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 229960001031 glucose Drugs 0.000 description 1
- 229960005150 glycerol Drugs 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000007902 hard capsule Substances 0.000 description 1
- 230000003862 health status Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 210000000087 hemolymph Anatomy 0.000 description 1
- 208000021760 high fever Diseases 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 239000011261 inert gas Substances 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 150000002596 lactones Chemical class 0.000 description 1
- 229960001375 lactose Drugs 0.000 description 1
- 229940124590 live attenuated vaccine Drugs 0.000 description 1
- 229940023012 live-attenuated vaccine Drugs 0.000 description 1
- 239000007937 lozenge Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 108010038320 lysylphenylalanine Proteins 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 239000000845 maltitol Substances 0.000 description 1
- VQHSOMBJVWLPSR-WUJBLJFYSA-N maltitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O VQHSOMBJVWLPSR-WUJBLJFYSA-N 0.000 description 1
- 235000010449 maltitol Nutrition 0.000 description 1
- 229940035436 maltitol Drugs 0.000 description 1
- 229940035034 maltodextrin Drugs 0.000 description 1
- 238000007726 management method Methods 0.000 description 1
- 229910052748 manganese Inorganic materials 0.000 description 1
- 229960001855 mannitol Drugs 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 208000015994 miscarriage Diseases 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000004223 monosodium glutamate Substances 0.000 description 1
- 210000000214 mouth Anatomy 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 210000003928 nasal cavity Anatomy 0.000 description 1
- 229960004927 neomycin Drugs 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 239000012457 nonaqueous media Substances 0.000 description 1
- 239000000346 nonvolatile oil Substances 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 150000002895 organic esters Chemical class 0.000 description 1
- 229940055726 pantothenic acid Drugs 0.000 description 1
- 235000019161 pantothenic acid Nutrition 0.000 description 1
- 239000011713 pantothenic acid 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
- 239000012188 paraffin wax Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 229920001983 poloxamer Polymers 0.000 description 1
- 229920000447 polyanionic polymer Polymers 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 229920000024 polymyxin B Polymers 0.000 description 1
- 229960005266 polymyxin b Drugs 0.000 description 1
- 229920005862 polyol Polymers 0.000 description 1
- 150000003077 polyols Chemical class 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 229940086065 potassium hydrogentartrate Drugs 0.000 description 1
- 235000010241 potassium sorbate Nutrition 0.000 description 1
- 239000004302 potassium sorbate Substances 0.000 description 1
- 229940069338 potassium sorbate Drugs 0.000 description 1
- 229920001592 potato starch Polymers 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000003380 propellant Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 229960004889 salicylic acid Drugs 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 235000017709 saponins Nutrition 0.000 description 1
- 239000004460 silage Substances 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 229940083542 sodium Drugs 0.000 description 1
- WXMKPNITSTVMEF-UHFFFAOYSA-M sodium benzoate Chemical compound [Na+].[O-]C(=O)C1=CC=CC=C1 WXMKPNITSTVMEF-UHFFFAOYSA-M 0.000 description 1
- 235000010234 sodium benzoate Nutrition 0.000 description 1
- 239000004299 sodium benzoate Substances 0.000 description 1
- 229960003885 sodium benzoate Drugs 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 229940045902 sodium stearyl fumarate Drugs 0.000 description 1
- 235000010265 sodium sulphite Nutrition 0.000 description 1
- 239000007901 soft capsule Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 229960002920 sorbitol Drugs 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 208000000995 spontaneous abortion Diseases 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- JBQYATWDVHIOAR-UHFFFAOYSA-N tellanylidenegermanium Chemical compound [Te]=[Ge] JBQYATWDVHIOAR-UHFFFAOYSA-N 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 229940033663 thimerosal Drugs 0.000 description 1
- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 210000004291 uterus Anatomy 0.000 description 1
- 210000001215 vagina Anatomy 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- MWOOGOJBHIARFG-UHFFFAOYSA-N vanillin Chemical compound COC1=CC(C=O)=CC=C1O MWOOGOJBHIARFG-UHFFFAOYSA-N 0.000 description 1
- FGQOOHJZONJGDT-UHFFFAOYSA-N vanillin Natural products COC1=CC(O)=CC(C=O)=C1 FGQOOHJZONJGDT-UHFFFAOYSA-N 0.000 description 1
- 235000012141 vanillin Nutrition 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 239000005723 virus inoculator Substances 0.000 description 1
- 235000019155 vitamin A Nutrition 0.000 description 1
- 239000011719 vitamin A Substances 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
- 239000011787 zinc oxide Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
- A61K2039/552—Veterinary vaccine
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/10011—Arteriviridae
- C12N2770/10034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Molecular Biology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Animal Husbandry (AREA)
- Epidemiology (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
본 발명은 돼지생식기호흡기증후군 바이러스 (PRRSV)의 T 세포 면역 유도 에피토프 펩타이드를 포함하는 면역원성 조성물에 관한 것으로, 본 발명의 펩타이드는 돼지생식기호흡기증후군 바이러스 (porcine reproductive and respiratory syndrome virus; PRRSV)에 감염된 돼지의 말초 혈액 단핵세포 (peripheral blood mononuclear cell; PBMC)를 자극하여 인터페론 감마의 발현을 유도하고, 타겟세포에서의 바이러스 증폭을 억제하여, 돼지생식기호흡기증후군 바이러스를 효과적으로 방어할 수 있다.The present invention relates to an immunogenic composition containing a T cell immunity-inducing epitope peptide of porcine reproductive and respiratory syndrome virus (PRRSV). It can effectively protect against the porcine reproductive and respiratory syndrome virus by stimulating porcine peripheral blood mononuclear cells (PBMC) to induce the expression of interferon gamma and inhibit virus amplification in target cells.
Description
본 발명은 PRRSV 유래 펩타이드를 포함하는 면역원성 조성물에 관한 것으로, 더욱 구체적으로 돼지생식기호흡기증후군 바이러스 (PRRSV)의 T 세포 면역 유도 에피토프 펩타이드를 포함하는 조성물에 관한 것이다.The present invention relates to an immunogenic composition containing a PRRSV-derived peptide, and more specifically to a composition containing a T cell immunity-inducing epitope peptide of porcine reproductive and respiratory syndrome virus (PRRSV).
돼지생식기호흡기증후군을 유발하는 돼지생식기호흡기증후군 바이러스 (porcine reproductive and respiratory syndrome virus; PRRSV)는 크게 북미형과 유럽형으로 나뉘며, 변이가 심해 수많은 종이 존재한다. 또한, 폐포 대식세포를 파괴하여 다른 병원체의 2차 감염이 일어나기 쉽고, 직접적으로 폐에 간질성 폐렴을 일으키며, 모돈에 급성 감염되면 고열을 동반하여 유산을 야기한다. 유산이 야기되지 않은 임신돈은 태반을 통해 PRRSV가 감염되어 태아가 폐사한다.Porcine reproductive and respiratory syndrome virus (PRRSV), which causes porcine reproductive and respiratory syndrome, is largely divided into North American and European types, and there are numerous species due to severe mutations. In addition, it destroys alveolar macrophages, making it easy for secondary infections with other pathogens to occur, directly causing interstitial pneumonia in the lungs, and acute infection in sows is accompanied by high fever and causes miscarriage. Pregnant sows that do not miscarry are infected with PRRSV through the placenta, resulting in fetal death.
돼지생식기호흡기증후군 바이러스는 돼지의 비강, 근육, 경구, 자궁, 그리고 질 등을 통해서 돼지생식기호흡기증후군을 유발한다. 일반적으로 돼지는 경구감염보다 비경구감염에 더 큰 감수성을 가진다. PRRSV 감염의 유형에는 비간접적인 전파, 수직 감염, 지속 감염, 농장 내에서의 전파, 농장 사이의 전파가 있다. 비간접적인 전파는 장비, 도구, 의복, 물, 음식, 매개곤충 (모기, 파리), 공기 중의 비말, 고기의 육즙 등을 통해서 일어날 수 있으며, PRRSV에 급성 감염된 돼지를 접촉한 후 60분 이내의 작업자의 작업복, 장화, 손 등에서도 PRRSV가 검출된다. 수직감염은 PRRSV가 바이러스 혈증을 보이는 임신 말기 모돈의 태반을 통해서 태아에게 전염되는 것을 의미한다.Porcine reproductive and respiratory syndrome virus causes porcine reproductive and respiratory syndrome through the nasal cavity, muscles, mouth, uterus, and vagina of pigs. In general, pigs are more susceptible to parenteral infections than oral infections. Types of PRRSV infection include indirect transmission, vertical transmission, persistent infection, transmission within farms, and transmission between farms. Indirect transmission can occur through equipment, tools, clothing, water, food, vector insects (mosquitoes, flies), droplets in the air, meat juices, etc., and can occur within 60 minutes after contact with a pig acutely infected with PRRSV. PRRSV is also detected in workers' work clothes, boots, and hands. Vertical transmission means that PRRSV is transmitted to the fetus through the placenta of a late-gestation sow with viremia.
또한, PRRSV는 돼지에서 만성적인 감염을 일으킨다. 바이러스는 수개월 동안 임상증상이 명확히 나타나지 않은 돼지의 감수성 세포 내에서도 복제된다. 이것은 PRRSV 전파의 가장 중요한 특징이며, 감염된 모돈이 99일까지도 바이러스를 전파시킬 수 있다고 보고되고 있다.Additionally, PRRSV causes chronic infections in pigs. The virus also replicates within susceptible cells in pigs that do not show clear clinical symptoms for several months. This is the most important characteristic of PRRSV transmission, and it has been reported that infected sows can transmit the virus for up to 99 days.
한편, 돼지생식기호흡기증후군의 원인체인 PRRSV를 발견한 이후 약 20년 동안 이의 예방법을 개발하기 위하여 많은 노력이 이어졌음에도 아직까지 효과적인 예방법 및 관리법이 개발되지 않은 실정이다. PRRSV 제어를 위해 불활화 백신과 약독화 생백신 등 다양한 백신이 개발되었으나 오직 감염성이 확보된 약독화 백신만이 만족스러운 수준의 방어효과를 유도하는 것으로 밝혀졌다. 하지만, PRRSV가 유전적으로 매우 다양하게 존재하기 때문에 다양한 바이러스간 교차면역의 부재로, 기존 상용 백신만으로 PRRSV의 감염을 완벽하게 방어하기 어려운 상황이다.Meanwhile, although much effort has been made to develop prevention methods for about 20 years since the discovery of PRRSV, the cause of porcine reproductive and respiratory syndrome, effective prevention and management methods have not yet been developed. Various vaccines, including inactivated vaccines and live attenuated vaccines, have been developed to control PRRSV, but only attenuated vaccines with guaranteed infectivity were found to induce a satisfactory level of protective effect. However, because PRRSV is genetically very diverse, it is difficult to completely protect against PRRSV infection with existing commercial vaccines due to the absence of cross-immunity between various viruses.
본 발명자는 돼지생식기호흡기증후군의 효과적인 예방을 위하여 연구하던 중, 본 발명자가 개발한 신규의 펩타이드가 T 세포 면역을 현저히 증가시킴으로써 PRRSV를 효과적으로 방어할 수 있음을 확인하고 본 발명을 완성하였다.While researching for effective prevention of porcine reproductive and respiratory syndrome, the present inventor confirmed that the novel peptide developed by the present inventor could effectively protect against PRRSV by significantly increasing T cell immunity, and completed the present invention.
따라서, 본 발명의 목적은 서열번호 1의 펩타이드 또는 서열번호 2의 펩타이드로 이루어진 돼지생식기호흡기증후군 바이러스 면역원성 펩티이드를 제공하는 것이다.Therefore, the object of the present invention is to provide a porcine reproductive and respiratory syndrome virus immunogenic peptide consisting of the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2.
또한, 본 발명의 또 다른 목적은 서열번호 1의 펩타이드 또는 서열번호 2의 펩타이드를 포함하는 면역원성 조성물, 돼지생식기호흡기증후군 예방 또는 치료용 약학 조성물 또는 사료 조성물을 제공하는 것이다.In addition, another object of the present invention is to provide an immunogenic composition containing the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2, a pharmaceutical composition for preventing or treating porcine reproductive and respiratory syndrome, or a feed composition.
또한, 본 발명의 또 다른 목적은 돼지에 서열번호 1의 펩타이드 또는 서열번호 2의 펩타이드를 투여하는 단계를 포함하는 돼지생식기호흡기증후군 예방 또는 치료방법을 제공하는 것이다.In addition, another object of the present invention is to provide a method for preventing or treating porcine reproductive and respiratory syndrome, which includes administering the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2 to a pig.
본 발명은 서열번호 1의 펩타이드 또는 서열번호 2의 펩타이드로 이루어진 돼지생식기호흡기증후군 바이러스 면역원성 펩타이드를 제공한다.The present invention provides a porcine reproductive and respiratory syndrome virus immunogenic peptide consisting of the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2.
본 발명은 서열번호 1의 펩타이드 또는 서열번호 2의 펩타이드를 포함하는 돼지생식기호흡기증후군 바이러스에 대한 면역원성 조성물을 제공한다.The present invention provides an immunogenic composition against porcine reproductive and respiratory syndrome virus comprising the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2.
본 발명은 서열번호 1의 펩타이드 또는 서열번호 2의 펩타이드를 포함하는 돼지생식기호흡기증후군 예방 또는 치료용 약학 조성물을 제공한다.The present invention provides a pharmaceutical composition for preventing or treating porcine reproductive and respiratory syndrome, comprising the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2.
본 발명은 서열번호 1의 펩타이드 또는 서열번호 2의 펩타이드를 포함하는 돼지생식기호흡기증후군 예방 또는 개선용 사료 조성물을 제공한다.The present invention provides a feed composition for preventing or improving porcine reproductive and respiratory syndrome, comprising the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2.
본 발명은 돼지에 서열번호 1의 펩타이드 또는 서열번호 2의 펩타이드를 투여하는 단계;를 포함하는 돼지생식기호흡기증후군 예방 또는 치료 방법을 제공한다.The present invention provides a method for preventing or treating porcine reproductive and respiratory syndrome, comprising administering the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2 to a pig.
본 발명의 펩타이드는 돼지생식기호흡기증후군 바이러스 (porcine reproductive and respiratory syndrome virus; PRRSV)에 감염된 돼지의 말초 혈액 단핵세포 (peripheral blood mononuclear cell; PBMC)를 자극하여 인터페론 감마의 발현을 유도하고, 타겟세포에서의 바이러스 증폭을 억제하여, 돼지생식기호흡기증후군 바이러스를 효과적으로 방어할 수 있다.The peptide of the present invention stimulates peripheral blood mononuclear cells (PBMC) of pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV) to induce the expression of interferon gamma in target cells. It can effectively protect against the porcine reproductive and respiratory syndrome virus by suppressing the amplification of the virus.
도 1은 돼지에서 실시예 1의 펩타이드 감염에 따른 IFN-γ유도능을 확인한 도이다.
도 2는 PRRSV의 증식 억제능 분석의 모식도이다.
도 3은 실시예 1의 펩타이드 혼합액 감염에 따른 바이러스 증식 억제능을 확인한 도이다.
도 4는 실시예 1의 펩타이드 개별 감염에 따른 바이러스 증식 억제능을 확인한 도이다.Figure 1 is a diagram confirming the IFN-γ inducing ability following infection with the peptide of Example 1 in pigs.
Figure 2 is a schematic diagram of the analysis of the proliferation inhibition ability of PRRSV.
Figure 3 is a diagram confirming the ability to inhibit virus proliferation following infection with the peptide mixture of Example 1.
Figure 4 is a diagram confirming the virus proliferation inhibition ability according to individual infection with the peptide of Example 1.
본 발명은 서열번호 1의 펩타이드 또는 서열번호 2의 펩타이드로 이루어진 돼지생식기호흡기증후군 바이러스 면역원성 펩타이드를 제공한다. The present invention provides a porcine reproductive and respiratory syndrome virus immunogenic peptide consisting of the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2.
본 발명의 돼지생식기호흡기증후군 바이러스 면역원성 펩타이드에 있어, 상기 펩타이드는 바람직하게는 국내바이러스 Type2 PRRSV 국내 바이러스를 기초로 합성되었으며 다른 서열로 이루어진 펩타이드 대비 면역 유도능이 현저히 우수한 T 세포 에피토프 펩타이드이다. 에피토프는 면역 체계에 의해 인식되는 항원상의 부위 (예컨대, 항체가 결합하는 부위)를 나타낸다. 에피토프는 하나 이상의 단백질의 삼차 접힘에 의해 병치된 연속 아미노산 또는 비연속 아미노산으로부터 형성될 수 있다. 에피토프는 전형적으로 적어도 3개, 보다 일반적으로는 적어도 5개 또는 8~10개의 아미노산을 독특한 공간적 형태에 포함한다.In the porcine reproductive and respiratory syndrome virus immunogenic peptide of the present invention, the peptide is preferably a T cell epitope peptide synthesized based on the domestic virus Type 2 PRRSV domestic virus and has a significantly superior immune inducing ability compared to peptides composed of other sequences. An epitope refers to a site on an antigen that is recognized by the immune system (e.g., a site to which an antibody binds). Epitopes can be formed from continuous or discontinuous amino acids juxtaposed by tertiary folding of one or more proteins. Epitopes typically contain at least 3, more commonly at least 5 or 8-10 amino acids in a unique spatial configuration.
본 발명의 펩타이드는 코딩된 또는 코딩되지 않은 아미노산 및 화학적으로 또는 생화학적으로 변형된 또는 유도체화된 아미노산을 포함하는, 임의의 길이의 아미노산의 중합체 형태를 나타낸다.Peptides of the invention represent polymeric forms of amino acids of any length, including encoded or non-encoded amino acids and amino acids that have been chemically or biochemically modified or derivatized.
본 발명의 용어 "면역원성"은 대상체에게 투여될 때 대상체에서 면역 반응을 이끌어내는 분자 (예컨대, 단백질, 핵산, 항원, 또는 유기체)의 선천적인 능력을 나타낸다. 이때, 항원은 본 발명에서 대상체와 접촉하게 될 때 (예컨대, 대상체에 존재하거나 대상체에 의해 검출될 때), 대상체로부터 검출 가능한 면역 반응을 초래하는 물질을 나타내기 위해 사용된다.The term “immunogenicity” herein refers to the innate ability of a molecule (e.g., protein, nucleic acid, antigen, or organism) to elicit an immune response in a subject when administered to the subject. At this time, antigen is used in the present invention to refer to a substance that, when brought into contact with a subject (e.g., present in or detected by the subject), results in a detectable immune response from the subject.
본 발명의 상기 펩타이드는 항원성 펩타이드라 할 수 있는데, 이는 대상체에 존재하거나 대상체에 의해 검출될 때 대상체에서 면역 반응의 설치(mounting)로 이어지는 펩타이드를 나타낸다.The peptide of the present invention may be referred to as an antigenic peptide, which refers to a peptide that, when present in or detected by a subject, leads to the mounting of an immune response in the subject.
또한, 본 발명은 서열번호 1의 펩타이드 또는 서열번호 2의 펩타이드를 포함하는 돼지생식기호흡기증후군 바이러스에 대한 면역원성 조성물을 제공한다. 이때, 상기 면역원성 조성물은, 바람직하게는 백신 또는 백신 조성물인 것이 좋다.Additionally, the present invention provides an immunogenic composition against porcine reproductive and respiratory syndrome virus comprising the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2. At this time, the immunogenic composition is preferably a vaccine or vaccine composition.
본 발명의 일 실시예에 의하면, 서열번호 1의 펩타이드 또는 서열번호 2의 펩타이드가 돼지에 접종시 T세포 면역을 현저히 증가시킴으로써 PRRSV를 효과적으로 방어하는데 이용할 수 있으며, PRRS 예방용 백신 혹은 백신 조성물로 유용하게 사용할 수 있음을 확인하였다.According to one embodiment of the present invention, the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2 can be used to effectively protect against PRRSV by significantly increasing T cell immunity when inoculated into pigs, and is useful as a vaccine or vaccine composition for preventing PRRS. It was confirmed that it can be used effectively.
본 발명의 펩타이드를 포함하는 백신 조성물은 불활화 바이러스를 이용한 백신과 비교하여, 제조공정 및 바이러스 접종이 편리하다는 우수성이 있다. PRRSV 는 MARC-145cell 에서 배양하여 사용하고 있으나, 바이러스 감염 4~5일 후 췌득시 바이러스 역가가 높지 않기 때문에 효과적인 역가를 포함하는 백신으로 제조하기 위해서는 대량으로 바이러스를 배양하고 바이러스 배양액를 울트라 센트리 퓨즈(30000RPM, 4H, 4도) 혹은 멤브레인을 이용하여 농축하는 과정이 필요한 단점이 있다. 또한 바이러스 선정과정에서 개발된 백신 균주가 국내 발생 바이러스와 매칭되어야 하기 때문에 바이러스를 선정하는데 어려움이 있었다. 반면 본 발명의 돼지생식기호흡기증후군 바이러스 면역원성 펩타이드를 이용하면, 대장균에서 생산하거나, 발현벡터 또는 감염성 벡터에 삽입하여 발현하는 방식으로 생산 할 수 있어 보다 용이하게 백신을 제조할 수 있고, 충분한 양의 백신을 효과적으로 제조할 수 있다. 또한 본 발명의 돼지생식기호흡기증후군 바이러스 면역원성 펩타이드는 Type 2 PRRSV 간의 멤브레인 서열 중에서 공통적인 서열로 만든 조합이므로, Type2 간의 PRRS 방어능도 가능한 장점이 있다. Compared to vaccines using inactivated viruses, the vaccine composition containing the peptide of the present invention has the advantage of being convenient for manufacturing processes and virus inoculation. PRRSV is used by culturing in MARC-145cell, but since the virus titer is not high when harvested 4-5 days after virus infection, in order to manufacture a vaccine with effective titer, the virus must be cultured in large quantities and the virus culture medium must be used in an ultra-centry fuse (30000RPM). , 4H, 4 degrees) or a concentration process using a membrane is required. In addition, there was difficulty in selecting the virus because the vaccine strain developed during the virus selection process had to match the domestically occurring virus. On the other hand, if the porcine reproductive and respiratory syndrome virus immunogenic peptide of the present invention is used, it can be produced in E. coli or by inserting into an expression vector or infectious vector, making it possible to manufacture a vaccine more easily and in sufficient quantity. Vaccines can be manufactured effectively. In addition, the porcine reproductive and respiratory syndrome virus immunogenic peptide of the present invention is a combination of sequences common among the membrane sequences between Type 2 PRRSV, so it has the advantage of being able to protect against Type 2 PRRSV.
본 발명의 백신은 추가로 약리학적으로 허용가능한 담체 또는 희석제를 포함한다. 백신에 적합한 담체는 기술분야의 당업자에게 공지되어 있으며, 단백질, 설탕 등을 포함하지만, 이에 한정되는 것은 아니다. 상기의 담체는 수용액 또는 비-수용액, 현탁액, 및 에멀전일 수 있다. 비-수용액 담체의 예는 프로필렌 글리콜, 폴리에틸렌 글리콜, 식용유 예컨대 올리브 오일 및 주사 가능한 유기 에스테르 예컨대 에틸 올리에이트를 들 수 있다. 수용액 담체는 식염수 및 완충배지를 포함하는, 물, 알콜/수용액, 에멀전 또는 현탁액을 포함한다. 비경구 담체는 염화나트륨 용액, 링거 덱스트로오스, 덱스트로오스 및 염화나트륨, 유산처리 링거 또는 고정 오일을 포함한다. 정맥주사용 담체는 예컨대 링거 덱스트로오스를 기본으로 하는 것과 같은 전해질 보충제, 액체 및 영양 보충제 등을 포함한다. 또한, 방부제 및 기타 첨가제 예컨대 항미생물제제, 항산화제, 킬레이트제, 불활성 가스 등이 추가로 존재할 수 있다. 바람직한 방부제로는 포르말린, 티메로살, 네오마이신, 폴리믹신 B 및 암포테리신 B를 포함할 수 있다.The vaccine of the present invention further comprises a pharmacologically acceptable carrier or diluent. Suitable carriers for vaccines are known to those skilled in the art and include, but are not limited to, proteins, sugars, etc. The carriers may be aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous carriers include propylene glycol, polyethylene glycol, edible oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcohol/aqueous solutions, emulsions or suspensions, including saline solutions and buffered media. Parenteral carriers include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's or fixed oils. Carriers for intravenous injection include electrolyte supplements, liquid and nutritional supplements, such as those based on Ringer's dextrose. In addition, preservatives and other additives such as antimicrobial agents, antioxidants, chelating agents, inert gases, etc. may additionally be present. Preferred preservatives may include formalin, thimerosal, neomycin, polymyxin B, and amphotericin B.
또한, 상기 백신 조성물은 어쥬번트를 추가로 포함할 수 있다.Additionally, the vaccine composition may further include an adjuvant.
상기 어쥬번트는 면역반응의 향상 및/또는 접종 후 흡수 속도를 촉진하는 화합물 또는 혼합물을 칭하는 것으로 임의의 흡수-촉진제를 포함한다. 허용가능한 어쥬번트는 프로인트 완전어쥬번트, 프로인트 불완전어쥬번트, 사포닌, 미네랄 젤 예컨대 수산화 알루미늄, 계면활성제, 리소레시틴, 플루론 폴리올, 다중음이온, 펩타이드, 오일, 탄화수소 에멀전, 키홀림펫 헤모시아닌, 디니트로페놀 등을 포함하나, 이에 제한되는 것은 아니다.The adjuvant refers to a compound or mixture that enhances the immune response and/or accelerates the rate of absorption after inoculation and includes an optional absorption-promoting agent. Acceptable adjuvants include Freund's complete adjuvant, Freund's incomplete adjuvant, saponins, mineral gels such as aluminum hydroxide, surfactants, lysolecithin, pluronic polyols, polyanions, peptides, oils, hydrocarbon emulsions, keyhole limpet hemolymph. It includes, but is not limited to, dinitrophenol, etc.
상기 백신은 생백신, 약독화 백신 및 사독 백신으로 이루어진 군에서 선택된 1종의 백신일 수 있다. 본 발명의 목적을 위한 백신조성물의 투여량은 체중 1kg 당 0.01㎖ 내지 1㎖를 투여하는 것이 바람직하다.The vaccine may be one type of vaccine selected from the group consisting of live vaccines, attenuated vaccines, and killed vaccines. The preferred dosage of the vaccine composition for the purpose of the present invention is 0.01 ml to 1 ml per 1 kg of body weight.
본 발명의 백신 조성물은 근육, 피하, 복강, 정맥, 경구, 진피, 안구 및 뇌로 이루어진 군에서 선택된 1종 이상의 투여 경로를 통해 투여하는 것이 바람직하다.The vaccine composition of the present invention is preferably administered through one or more administration routes selected from the group consisting of muscle, subcutaneous, abdominal, intravenous, oral, dermal, eye, and brain.
본 발명에 있어, 상기 면역원성 조성물은, 바람직하게는 T 세포에서 인터페론-감마 (INF-γ)의 발현을 유도하는 것이 좋고, 불활화 백신보다 돼지생식기호흡기증후군 바이러스(PRRSV) 증식억제능이 높은 것이 좋다.In the present invention, the immunogenic composition preferably induces the expression of interferon-gamma (INF-γ) in T cells and has a higher ability to inhibit the proliferation of porcine reproductive and respiratory syndrome virus (PRRSV) than an inactivated vaccine. good night.
본 발명의 일 실시예에 의하면, PRRSV로 감염된 돼지에서 분리한 T 세포를 본 펩타이드로 자극한 경우, INF-γ의 발현을 유도하였고, 자극된 T 세포는 타켓에서 감염된 바이러스의 성장을 억제하였음을 확인하였다.According to one embodiment of the present invention, when T cells isolated from pigs infected with PRRSV were stimulated with this peptide, the expression of INF-γ was induced, and the stimulated T cells inhibited the growth of the virus infected in the target. Confirmed.
한편, 본 발명은 서열번호 1의 펩타이드 또는 서열번호 2의 펩타이드를 포함하는 돼지생식기호흡기증후군 예방 또는 치료용 약학 조성물을 제공한다.Meanwhile, the present invention provides a pharmaceutical composition for preventing or treating porcine reproductive and respiratory syndrome, comprising the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2.
본 발명에서 사용되는 용어, "예방"이란 본 발명에 따른 약학적 조성물의 투여에 의해 돼지생식기호흡기증후군을 억제시키거나 발병을 지연시키는 모든 행위를 의미한다.As used in the present invention, the term “prevention” refers to all actions that suppress or delay the onset of porcine reproductive and respiratory syndrome by administering the pharmaceutical composition according to the present invention.
본 발명에서 사용되는 용어, "치료"란 본 발명에 따른 약학적 조성물의 투여에 의해 돼지생식기호흡기증후군에 의한 증세가 호전되거나 이롭게 변경되는 모든 행위를 의미한다.As used in the present invention, the term “treatment” refers to any action in which symptoms caused by porcine reproductive and respiratory syndrome are improved or beneficially changed by administration of the pharmaceutical composition according to the present invention.
본 발명의 상기 펩타이드를 포함하는 조성물은 상기 성분에 추가로 동일 또는 유사한 기능을 나타내는 유효성분을 1종 이상 함유할 수 있다. 또한, 본 발명의 약학적 조성물은 본 발명의 펩타이드 이외에 약학적으로 허용 가능한 담체를 추가로 포함할 수 있다.The composition containing the peptide of the present invention may contain one or more active ingredients that exhibit the same or similar functions in addition to the above ingredients. Additionally, the pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier in addition to the peptide of the present invention.
본 발명에서 사용될 수 있는 담체의 종류는 특별히 제한되지 아니하며 당해 기술 분야에서 통상적으로 사용되는 담체라면 어느 것이든 사용할 수 있다. 상기 담체의 비제한적인 예로는, 식염수, 멸균수, 링거액, 완충 식염수, 알부민 주사 용액, 락토오스, 덱스트로오스, 수크로오스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 말토덱스트린, 글리세롤, 에탄올 등을 들 수 있다. 이들은 단독으로 사용되거나 2종 이상을 혼합하여 사용될 수 있다.The type of carrier that can be used in the present invention is not particularly limited, and any carrier commonly used in the art can be used. Non-limiting examples of the carrier include saline solution, sterile water, Ringer's solution, buffered saline solution, albumin injection solution, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, maltodextrin, glycerol, ethanol, etc. You can. These may be used alone or in combination of two or more types.
또한, 본 발명의 약학적 조성물은 필요한 경우, 부형제, 희석제, 항산화제, 완충액 또는 정균제 등 기타 약학적으로 허용 가능한 첨가제들을 첨가하여 사용할 수 있으며, 충진제, 증량제, 습윤제, 붕해제, 분산제, 계면 활성제, 결합제 또는 윤활제 등을 부가적으로 첨가하여 사용할 수 있다.In addition, the pharmaceutical composition of the present invention can be used by adding other pharmaceutically acceptable additives, such as excipients, diluents, antioxidants, buffers or bacteriostatic agents, if necessary, as well as fillers, extenders, wetting agents, disintegrants, dispersants, and surfactants. , it can be used by additionally adding a binder or lubricant.
본 발명의 약학적 조성물에 있어서, 상기 본 발명의 펩타이드는 약학적 조성물의 전체의 중량을 기준으로 0.00001중량％ 내지 99.99중량％로 포함될 수 있으며, 바람직하게는 0.1중량% 내지 90중량%, 보다 바람직하게는 0.1중량% 내지 70중량%, 더욱 바람직하게는 0.1중량% 내지 50중량%로 포함될 수 있으나, 이에 한정되지 않으며 투여 대상의 상태, 병증의 진행 정도 등에 따라 다양하게 변경될 수 있다. 필요한 경우, 약학적 조성물의 전체 함량으로도 포함될 수 있다.In the pharmaceutical composition of the present invention, the peptide of the present invention may be included in an amount of 0.00001% to 99.99% by weight, preferably 0.1% by weight to 90% by weight, more preferably 0.1% by weight to 90% by weight, based on the total weight of the pharmaceutical composition. Typically, it may be included in an amount of 0.1% to 70% by weight, more preferably 0.1% to 50% by weight, but is not limited thereto and may vary depending on the condition of the administration subject, the degree of disease progression, etc. If necessary, it may also be included in the total content of the pharmaceutical composition.
본 발명의 약학적 조성물은 경구 투여 또는 비경구 투여를 위한 적합하고 다양한 제형으로 제제화되어 사용될 수 있다.The pharmaceutical composition of the present invention can be formulated and used in various suitable dosage forms for oral or parenteral administration.
본 발명의 약학적 조성물을 이용한 경구 투여용 제제의 비제한적인 예로는, 트로키제(troches), 로젠지(lozenge), 정제, 수용성 현탁액, 유성 현탁액, 조제 분말, 과립, 에멀젼, 하드 캡슐, 소프트 캡슐, 시럽 또는 엘릭시르제 등을 들 수 있다.Non-limiting examples of preparations for oral administration using the pharmaceutical composition of the present invention include troches, lozenges, tablets, aqueous suspensions, oily suspensions, preparation powders, granules, emulsions, hard capsules, and soft capsules. Capsules, syrups, or elixirs may be used.
본 발명의 약학적 조성물을 경구 투여용으로 제제화하기 위하여, 락토오스, 사카로오스, 솔비톨, 만니톨, 전분, 아밀로펙틴, 셀룰로오스 또는 젤라틴 등과 같은 결합제; 디칼슘 포스페이트 등과 같은 부형제; 옥수수 전분 또는 고구마 전분 등과 같은 붕해제; 스테아르산 마그네슘, 스테아르산 칼슘, 스테아릴 푸마르산 나트륨 또는 폴리에틸렌 글리콜 왁스 등과 같은 윤활유 등을 사용할 수 있으며, 감미제, 방향제, 시럽제 등도 사용할 수 있다. 나아가 캡슐제의 경우에는 상기 언급한 물질 외에도 지방유와 같은 액체 담체 등을 추가로 사용할 수 있다.In order to formulate the pharmaceutical composition of the present invention for oral administration, binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin; excipients such as dicalcium phosphate; disintegrants such as corn starch or sweet potato starch; Lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate, or polyethylene glycol wax can be used, and sweeteners, fragrances, syrups, etc. can also be used. Furthermore, in the case of capsules, in addition to the above-mentioned substances, a liquid carrier such as fatty oil can be additionally used.
본 발명의 약학적 조성물을 이용한 비경구용 제제의 비제한적인 예로는, 주사액, 좌제, 호흡기 흡입용 분말, 스프레이용 에어로졸제, 연고, 도포용 파우더, 오일, 크림 등을 들 수 있다.Non-limiting examples of parenteral preparations using the pharmaceutical composition of the present invention include injections, suppositories, powders for respiratory inhalation, aerosols for sprays, ointments, powders for application, oils, creams, etc.
본 발명의 약학적 조성물을 비경구 투여용으로 제제화하기 위하여, 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결 건조 제제, 외용제 등을 사용할 수 있으며, 상기 비수성용제, 현탁제로는 프로필렌글리콜, 폴리에틸렌글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다.To prepare the pharmaceutical composition of the present invention for parenteral administration, sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, topical preparations, etc. can be used. Examples of the non-aqueous solvents and suspensions include propylene glycol and polyethylene. Glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, etc. may be used.
본 발명의 약학적 조성물을 주사액으로 제제화하는 경우, 본 발명의 약학적 조성물을 안정제 또는 완충제와 함께 물에서 혼합하여 용액 또는 현탁액으로 제조하고 이를 앰플(ampoule) 또는 바이알(vial)의 단위 투여용으로 제제화할 수 있다.When the pharmaceutical composition of the present invention is formulated as an injection solution, the pharmaceutical composition of the present invention is mixed in water with a stabilizer or buffer to prepare a solution or suspension, which is then administered in ampoules or vials for unit administration. It can be formulated.
본 발명의 약학적 조성물을 에어로졸제로 제제화하는 경우, 수분산된 농축물 또는 습윤 분말이 분산되도록 추진제 등이 첨가제와 함께 배합될 수 있다.When the pharmaceutical composition of the present invention is formulated as an aerosol, propellants, etc. may be mixed with additives to disperse the water-dispersed concentrate or wet powder.
본 발명의 약학적 조성물을 연고, 크림, 도포용 파우더, 오일, 피부 외용제 등으로 제제화하는 경우에는, 동물성 유, 식물성 유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크, 산화 아연 등을 담체로 사용하여 제제화할 수 있다.When the pharmaceutical composition of the present invention is formulated into ointments, creams, powders for application, oils, external skin preparations, etc., animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite , silica, talc, zinc oxide, etc. can be used as carriers.
본 발명의 약학적 조성물의 약학적 유효량, 유효 투여량은 약학적 조성물의 제제화 방법, 투여 방식, 투여 시간 및/또는 투여 경로 등에 의해 다양해질 수 있으며, 약학적 조성물의 투여로 달성하고자 하는 반응의 종류와 정도, 투여 대상이 되는 개체의 연령, 체중, 일반적인 건강 상태, 질병의 증세나 정도, 성별, 식이, 배설, 해당 개체에 동시 또는 이시에 함께 사용되는 약물 기타 조성물의 성분 등을 비롯한 여러 인자 및 의약 분야에서 잘 알려진 유사 인자에 따라 다양해질 수 있으며, 당해 기술 분야에서 통상의 지식을 가진 자는 목적하는 치료에 효과적인 투여량을 용이하게 결정하고 처방할 수 있다. 예를 들어, 본 발명의 약학적 조성물의 일일 투여량은 약 0.01 내지 1,000mg/kg이고, 바람직하게는 0.1 내지 100mg/kg이며, 하루 일회 내지 수회에 나누어 투여할 수 있다.The pharmaceutically effective amount and effective dosage of the pharmaceutical composition of the present invention may vary depending on the formulation method, administration method, administration time, and/or administration route of the pharmaceutical composition, and the reaction to be achieved by administration of the pharmaceutical composition may be determined. Various factors including the type and extent, age, weight, general health status, symptoms or severity of disease, gender, diet, excretion, drugs used simultaneously or simultaneously with the subject, and other composition components, etc. of the subject to be administered. and similar factors well known in the pharmaceutical field, and a person skilled in the art can easily determine and prescribe an effective dosage for the desired treatment. For example, the daily dosage of the pharmaceutical composition of the present invention is about 0.01 to 1,000 mg/kg, preferably 0.1 to 100 mg/kg, and can be administered once or in divided doses several times a day.
한편, 본 발명은 서열번호 1의 펩타이드 또는 서열번호 2의 펩타이드를 포함하는 돼지생식기호흡기증후군 예방 또는 개선용 사료 조성물을 제공한다.Meanwhile, the present invention provides a feed composition for preventing or improving porcine reproductive and respiratory syndrome comprising the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2.
본 발명의 상기 사료 조성물은 발효 사료, 배합 사료, 펠렛 형태 및 사일리지(silage) 등의 형태로 제조될 수 있으나, 이에 한정되는 것은 아니고, 필요에 따라 변형하여 제조할 수 있다.The feed composition of the present invention may be manufactured in the form of fermented feed, compounded feed, pellet form, silage, etc., but is not limited thereto, and may be modified and manufactured as necessary.
본 발명의 사료 조성물은 통상 사용되어 냄새, 맛, 시각 등을 향상시킬 수 있는 추가 성분을 포함할 수 있다. 예들 들어, 비타민 A, C, D, E, B1, B2, B6, B12, 니아신(niacin), 비오틴(biotin), 폴레이트(folate), 판토텐산(panthotenic acid) 등을 포함할 수 있다. 또한, 아연(Zn), 철(Fe), 칼슘(Ca), 크롬(Cr), 마그네슘(Mg), 망간(Mn), 구리(Cu) 등의 미네랄을 포함할 수 있다. 또한, 라이신, 트립토판, 시스테인, 발린 등의 아미노산을 포함할 수 있다. 또한, 방부제(소르빈산 칼륨, 벤조산나트륨, 살리실산, 디히드로초산나트륨 등), 살균제(표백분과 고도 표백분, 차아염소산나트륨 등), 산화방지제(부틸히드록시아니졸(BHA), 부틸히드록시톨루엔(BHT) 등), 착색제(타르색소 등), 발색제(아질산 나트륨, 아초산 나트륨 등), 표백제(아황산나트륨), 조미료(MSG 글루타민산나트륨 등), 감미료(둘신, 사이클레메이트, 사카린, 나트륨 등), 향료(바닐린, 락톤류 등), 팽창제(명반, D-주석산수소칼륨 등), 강화제, 유화제, 증점제(호료), 피막제, 검기초제, 거품억제제, 용제, 개량제 등의 식품 첨가물(food additives)을 첨가할 수 있다. 상기 첨가물은 적절한 양으로 사용될 수 있다.The feed composition of the present invention may contain additional ingredients that are commonly used to improve smell, taste, vision, etc. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. Additionally, it may contain minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), and copper (Cu). Additionally, it may contain amino acids such as lysine, tryptophan, cysteine, and valine. In addition, preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dihydroacetate, etc.), disinfectants (bleaching powder, high bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxytoluene (BHT) ), etc.), colorants (tar colors, etc.), coloring agents (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG monosodium glutamate, etc.), sweeteners (dulcine, cyclemate, saccharin, sodium, etc.), Food additives such as flavorings (vanillin, lactones, etc.), leavening agents (alum, D-potassium hydrogen tartrate, etc.), strengthening agents, emulsifiers, thickeners (grease), coating agents, gum base agents, anti-foam agents, solvents, improvers, etc. can be added. The above additives can be used in appropriate amounts.
본 발명의 사료 조성물에 있어서, 본 발명 펩타이드의 함량은 특별히 제한되지 않으며, 투여 대상의 상태, 병증의 진행 정도 등에 따라 다양하게 변경될 수 있다.In the feed composition of the present invention, the content of the peptide of the present invention is not particularly limited and may vary depending on the condition of the administration subject, the progress of the disease, etc.
한편, 본 발명은 돼지에 서열번호 1의 펩타이드 또는 서열번호 2의 펩타이드를 투여하는 단계;를 포함하는 돼지생식기호흡기증후군 예방 또는 치료 방법을 제공한다.Meanwhile, the present invention provides a method for preventing or treating porcine reproductive and respiratory syndrome, comprising administering the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2 to a pig.
본 발명에서 사용된 용어, "투여"는 어떠한 적절한 방법으로 돼지에게 본 발명의 펩타이드를 도입하는 것을 의미하며, 본 발명의 조성물의 투여 경로는 목적 조직에 도달할 수 있는 한 경구 또는 비경구의 다양한 경로를 통하여 투여될 수 있다.As used in the present invention, the term "administration" means introducing the peptide of the present invention into a pig by any suitable method, and the route of administration of the composition of the present invention is various oral or parenteral routes as long as it can reach the target tissue. It can be administered through.
본 발명 펩타이드의 비경구 투여 방법으로는, 정맥 내 투여, 복강 내 투여, 근육 내 투여, 경피 투여 또는 피하 투여 등을 이용할 수 있으며, 상기 펩타이드를 분무, 흡입하는 방법 또한 이용할 수 있으나 이에 제한되지 않는다.Parenteral administration methods for the peptide of the present invention include intravenous administration, intraperitoneal administration, intramuscular administration, transdermal administration, or subcutaneous administration. Methods of spraying or inhaling the peptide may also be used, but are not limited thereto. .
본 발명의 펩타이드의 투여는 하루에 1회 투여될 수 있고, 수회에 나누어 투여될 수도 있다. 본 발명의 펩타이드는 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와는 순차적 또는 동시에 투여될 수 있다. 상기 요소를 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양으로 투여할 수 있으며, 이는 당업자에 의해 용이하게 결정될 수 있다.The peptide of the present invention may be administered once a day, or may be administered in several divided doses. The peptide of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents. Considering all of the above factors, it can be administered in an amount that can achieve maximum effect with the minimum amount without side effects, and this can be easily determined by a person skilled in the art.
본 발명의 펩타이드의 투여 경로 및 투여 방식은 각각 독립적일 수 있으며, 목적하는 타겟에 상기 펩타이드가 도달할 수 있는 한, 특별한 제한 없이 임의의 투여 경로 및 투여 방식에 따를 수 있다.The administration route and administration method of the peptide of the present invention may be independent, and any administration route and administration method may be used without particular limitation as long as the peptide can reach the intended target.
본 발명의 펩타이드는 돼지생식기호흡기증후군을 예방, 개선 또는 치료하기 위하여 추가적으로 호르몬 치료, 약물 치료 등의 다양한 방법들과 병용하여 사용될 수 있다.The peptide of the present invention can be used in combination with various methods such as hormone treatment and drug treatment to prevent, improve, or treat porcine reproductive and respiratory syndrome.
이하, 본 발명의 내용을 하기 실시예 또는 실험예를 통해 더욱 상세히 설명하기로 한다. 다만, 본 발명의 권리범위가 하기 실시예 및 실험예에만 한정되는 것은 아니고, 그와 등가의 기술적 사상의 변형까지를 포함한다.Hereinafter, the contents of the present invention will be described in more detail through the following examples or experimental examples. However, the scope of the present invention is not limited to the following examples and experimental examples, and includes modifications of the technical idea equivalent thereto.
[[ 실시예Example 1: 본 발명의 1: of the present invention 펩타이드peptide 선발] Selection]
돼지생식기호흡기증후군 바이러스를 방어하기 위한 펩타이드를 선발하기 위하여, 국내바이러스 Type2 PRRSV의 막 단백질의 아미노산 서열을 기초로 유전자 상동성이 100% 유사한 T세포 에피토프 펩타이드 10종을 선정 및 합성하였고 이들의 Spanning region 정보를 분석한 결과를 표 1에 나타내었다. 선정된 T세포 에피토프 펩타이드는 20 내지 38개 아미노산 서열로 이루어져 있으며, 이하 실험에서 이들 10종에 대한 비교 실험을 수행하였다. In order to select peptides to protect against the porcine reproductive and respiratory syndrome virus, 10 types of T cell epitope peptides with 100% similar genetic homology were selected and synthesized based on the amino acid sequence of the membrane protein of the domestic virus Type 2 PRRSV, and their spanning region was selected. The results of analyzing the information are shown in Table 1. The selected T cell epitope peptide consists of 20 to 38 amino acid sequences, and a comparative experiment was performed on these 10 types in the following experiment.
[[ 실험예Experiment example 1: One: 실시예Example 1의 1's 펩타이드에To peptides 의한 세포면역 증가 확인] [Confirmation of increase in cellular immunity]
상시 실시예 1을 통해 선발된 10종 펩타이드의 IFN-γ유도능을 분석하기 위해, ELISpot (Enzyme linked immunospot) kit (MABTECH)를 실시하였다. 3주령(개체번호 #90, #93)의 돼지에 국내 분리주 (Type 2 PRRS LMY, Accession No. GenBank accession no.DQ473474.1)를 (근육접종: 104. 5TCID50/ml)감염시키고 5주 후 돼지에서 얻은 PBMC를 세포 면역능 확인 실험에 이용하였다. 개체번호 #90, #93의 각각의 PBMC가 1x106cells/well 포함되도록 웰을 96well U form plate에 준비 하고 항원자극을 위한 물질로 10종의 펩타이드 각 10 μM/well 또는 불활화된 LMY (PRRS LMY 모균주 (Accession No. GenBank accession no.DQ473474.1, MARC-145 cell 에서 7번 계대 배양 하여 얻은 바이러스 주) 1x106/well 와 양성 대조군인 PHA 0.5μg/well, 음성대조군인 무혈청 배지를 각각 넣고 PBMC와 항원을 파이펫을 이용하여 섞어 반응 시킨 후 이전에 capture mAb가 코팅된 ELISA plate 에 옮겨주고, 5% CO2, 37℃에서 21시간 반응시켰다. 그 후, biotin이 표지된 anti-IFN-gamma 1차 항체 및 streptavidin-HRP가 표지된 2차 항체 (상기 항체들은 모두 ELISpot kit (MABTECH)에 포함된 것임)를 순차적으로 처리하고, TMB발색 후 세포 1x106개 당 spot forming unit (SFU)를 계산하였다. 이 때, SFU 수치는 IFN- γ 발현수준을 나타내며, 각 실험군의 IFN- γ 분비량을 도 1 에 나타내었다. To analyze the IFN-γ inducing ability of the 10 peptides selected through Example 1, an ELISpot (Enzyme linked immunospot) kit (MABTECH) was performed. Pigs aged 3 weeks (individual numbers #90, #93) were infected with a domestic isolate (Type 2 PRRS LMY, Accession No. GenBank accession no.DQ473474.1) (intramuscular inoculation: 10 4. 5 TCID 50 /ml) and 5 PBMCs obtained from pigs after one week were used in experiments to confirm cellular immunity. Wells were prepared in a 96-well U form plate to contain 1x10 6 cells/well of each PBMC of individual numbers #90 and #93, and 10 types of peptides were added at 10 μM/well each or inactivated LMY (PRRS) as a substance for antigen stimulation. LMY parent strain (Accession No. GenBank accession no.DQ473474.1, virus strain obtained by subculturing 7 times in MARC-145 cells) 1x10 6 /well, PHA 0.5μg/well as positive control, and serum-free medium as negative control. Each was added, PBMC and antigen were mixed using a pipette, transferred to an ELISA plate previously coated with capture mAb, and reacted for 21 hours at 37°C with 5% CO 2. Afterwards, biotin-labeled anti- The primary antibody for IFN-gamma and the secondary antibody labeled with streptavidin-HRP (all of the above antibodies are included in the ELISpot kit (MABTECH)) were sequentially treated, and after TMB staining, spot forming unit (SFU) was added per 1x106 cells. ) was calculated. At this time, the SFU value represents the level of expression of IFN- γ, and the amount of IFN- γ secretion in each experimental group is shown in Figure 1.
도 1에 나타낸 바와 같이, 개체 #90번, #93번 PBMC에서 펩타이드 처리군에 따라IFN- γ 분비량은 각각 상이하게 나타났다. 특히 PS7번 PS8번 처리 그룹이 양성 컨트롤인 불활화 LMY 항원 처리군과 비교 하였을 때 평균 이상이거나 유사한 수준으로 IFN- γ가 분비됨을 확인하였으나, 나머지 펩타이드 서열에서는 IFN-γ발현이 음성 대조군과 같거나 수치가 낮음을 확인하였다. 이러한 결과는 모든 막 단백질 펩타이드 서열 기초 T 세포 에피토프 펩타이드들이 면역원성 유도 효과를 나타내는 것이 아니며, 특정 서열을 가진 펩타이드만이 효과적으로 면역원성 유도 효과를 나타냄을 보여주는 결과이다. 가장 우수한 효과를 나타낸 PS7번 PS8번 펩타이드의 서열 정보를 하기 표 2에 나타내었다. As shown in Figure 1, the amount of IFN-γ secretion was different depending on the peptide treatment group in individual #90 and #93 PBMC. In particular, it was confirmed that the PS7 and PS8 treatment groups secreted IFN-γ at a level that was above or similar to the average when compared to the inactivated LMY antigen treatment group, which was a positive control. However, in the remaining peptide sequences, the expression of IFN-γ was the same as that of the negative control group. It was confirmed that the value was low. These results show that not all membrane protein peptide sequence-based T cell epitope peptides exhibit an immunogenicity-inducing effect, and only peptides with a specific sequence effectively exhibit an immunogenicity-inducing effect. The sequence information of the PS7 and PS8 peptides that showed the best effect are shown in Table 2 below.
[[ 실험예Experiment example 2: 2: 실시예Example 1의 1's 펩타이드의of peptides 바이러스 증식 virus growth 억제능inhibitory ability 확인] check]
2.1 2.1 실시예Example 1의 1's 펩타이드peptide 혼합액의 바이러스 증식 Virus growth in mixed solution 억제능inhibitory ability 평가 evaluation
국내분리주(LMY: Type 2 PRRS LMY, Accession No. GenBank accession no.DQ473474.1)의 돼지감염 후 5주째에 추출된 PBMC를 선발된 실시예 1의 10종의 펩타이드의 혼합액 혹은 불활화 항원(heat inactivated LMY, no.DQ473474.1)으로 자극하였다. 불활화 항원인 heat inactivated LMY 는 1x107/ml을 56℃, 30분 열처리하여 불활화처리된 항원이며 이를 PBMC에 10μl를 처리하여 PBMC를 자극 하였다. 펩타이드 혼합액은 선별된 실시예 1의 펩타이드 10종을 1uM씩 혼합하여 혼합액 10uM을 만드는 방법으로 제조하였으며, 제조된 펩타이드 혼합액으로 PBMC를 자극시켰다. 무혈청배지를 PBMC에 첨가하여 음성대조군을 설정하였다. 자극 7일 뒤, 자극된 말초 혈액 단핵세포와 감염성 PRRS 바이러스 (live LMY, 0.00005moi)와 타겟세포인 단핵 세포 유도 대식세포 (monocyte-derived macrophage, MDM)를 4일 동안 공배양하였다. 이후, LMY 바이러스 증식 억제작용 여부를 실시간 중합효소연쇄반응 (realtime PCR)을 이용하여 분석하였으며, 해당 처리과정을 도 2에 PCR 결과를 도 3에 나타내었다. PBMCs extracted 5 weeks after infection of pigs with a domestic isolate (LMY: Type 2 PRRS LMY, Accession No. GenBank accession no.DQ473474.1) were treated with a mixture of the 10 peptides of Example 1 or inactivated antigen (heat). Stimulation was performed with inactivated LMY, no.DQ473474.1). Heat inactivated LMY, an inactivated antigen, is an antigen that was inactivated by heat treatment at 56°C for 30 minutes at 1x10 7 /ml, and 10 μl of this was treated with PBMC to stimulate PBMC. The peptide mixture was prepared by mixing 10 uM of the selected 10 peptides from Example 1 to make 10uM of the mixture, and PBMCs were stimulated with the prepared peptide mixture. A negative control group was set up by adding serum-free medium to PBMC. Seven days after stimulation, stimulated peripheral blood mononuclear cells were co-cultured with infectious PRRS virus (live LMY, 0.00005 moi) and monocyte-derived macrophages (MDM), which were target cells, for 4 days. Afterwards, the inhibitory effect of LMY virus proliferation was analyzed using real-time polymerase chain reaction (realtime PCR), and the processing process is shown in Figure 2 and the PCR results are shown in Figure 3.
도 3에 나타낸 바와 같이, 실험결과 10종의 펩타이드가 혼합된 혼합액은 동일 열 불활화 바이러스 보다 더 높은 수준의 바이러스 증식 억제 능력을 보여줌을 확인하였다. 구체적으로 열 불활화 바이러스로 자극된 실험군 (HIV 자극) 에서는 음성대조군(PBMC 비 자극)에 비하여 80.4% 억제능을 확인하였으나, 본 발명의 펩타이드 혼합액은 바이러스 증식을 97%까지 억제하였음을 확인하였다. As shown in Figure 3, the experimental results confirmed that the mixture of 10 types of peptides showed a higher level of virus proliferation inhibition ability than the same heat-inactivated virus. Specifically, in the experimental group stimulated with heat-inactivated virus (HIV stimulation), 80.4% inhibition was confirmed compared to the negative control group (PBMC non-stimulation), but the peptide mixture of the present invention was confirmed to inhibit virus proliferation by up to 97%.
2.2 2.2 실시예Example 1의 1's 펩타이드의of peptides 바이러스 증식 virus proliferation 억제능inhibitory ability 평가 evaluation
상기 실험을 통해 펩타이드 혼합액에서 바이러스 증식 억제능력이 불활화된 바이러스에 비해 더욱 높게 나타난 것을 확인하였고, 개별 펩타이드도 실질적인 바이러스 증식 억제능을 가지는지 여부를 접종 이후 작출되는 바이러스의 양을 TCID50으로 측정하여 확인하였다. 개별 펩타이드를 10uM로 각각 자극한 것을 제외하고는 상기 실험예 2.1의 펩타이드 혼합액과 동일한 방법으로 바이러스 증식 억제능 평가를 수행하였으며, 그 결과를 도 4에 나타내었다. Through the above experiment, it was confirmed that the virus growth inhibition ability in the peptide mixture was higher than that of the inactivated virus, and whether individual peptides also had a substantial virus growth inhibition ability was confirmed by measuring the amount of virus produced after inoculation with TCID50. did. The virus proliferation inhibitory ability was evaluated in the same manner as the peptide mixture in Experimental Example 2.1, except that each peptide was stimulated with 10uM, and the results are shown in Figure 4.
도 4에 나타낸 바와 같이, 실시예 1의 펩타이드 중 PS7의 바이러스 증식억제율이 98.1%로 확인되었고, PS8의 바이러스 증식억제율이 99.6%로 확인되어, 타겟 세포에서 높은 수준, 즉 불활화바이러스와 유사한 수준의 바이러스 증식 억제능을 가짐을 확인하였다. 한편, 그 외 펩타이드는 약한 수준의 바이러스 억제능을 보여 실험예 1의 IFN- γ 분비 효과와 일치하는 결과를 확인하였다. As shown in Figure 4, among the peptides of Example 1, the virus proliferation inhibition rate of PS7 was confirmed to be 98.1%, and the virus proliferation inhibition rate of PS8 was confirmed to be 99.6%, showing a high level in target cells, that is, a level similar to that of the inactivated virus. It was confirmed to have the ability to inhibit virus proliferation. Meanwhile, other peptides showed a weak level of virus inhibition, a result consistent with the effect of IFN-γ secretion in Experimental Example 1.
상기와 같은 결과를 통해, 펩타이드 PS7 및 PS8 이 PRRS 바이러스 증식 억제 효과가 더욱 우수하고 면역유도 효과가 우수함을 확인하였으며, 이들 펩타이드들이 불활화 바이러스를 대체하여 백신으로 활용가능함을 확인하였다. Through the above results, it was confirmed that peptides PS7 and PS8 had a better effect of inhibiting the proliferation of PRRS virus and had an excellent immune inducing effect, and it was confirmed that these peptides can be used as a vaccine by replacing inactivated viruses.
<110> REPUBLIC OF KOREA(Animal and Plant Quarantine Agency) <120> Composition for antigenicity comprising peptide from PRRSV <130> 1-107P <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 26 <212> PRT <213> porcine reproductive and respiratory syndrome virus <400> 1 Leu Leu Ala Phe Ser Ile Thr Tyr Thr Pro Val Met Ile Tyr Ala Leu 1 5 10 15 Lys Val Ser Arg Gly Arg Leu Leu Gly Leu 20 25 <210> 2 <211> 38 <212> PRT <213> porcine reproductive and respiratory syndrome virus <400> 2 Leu Trp Gly Val Tyr Ser Ala Ile Glu Thr Trp Lys Phe Ile Thr Ser 1 5 10 15 Arg Cys Arg Leu Cys Leu Leu Gly Arg Lys Tyr Ile Leu Ala Pro Ala 20 25 30 His His Val Glu Ser Ala 35 <110> REPUBLIC OF KOREA(Animal and Plant Quarantine Agency) <120> Composition for antigenicity comprising peptide from PRRSV <130> 1-107P <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 26 <212> PRT <213> porcine reproductive and respiratory syndrome virus <400> 1 Leu Leu Ala Phe Ser Ile Thr Tyr Thr Pro Val Met Ile Tyr Ala Leu 1 5 10 15 Lys Val Ser Arg Gly Arg Leu Leu Gly Leu 20 25 <210> 2 <211> 38 <212> PRT <213> porcine reproductive and respiratory syndrome virus <400> 2 Leu Trp Gly Val Tyr Ser Ala Ile Glu Thr Trp Lys Phe Ile Thr Ser 1 5 10 15 Arg Cys Arg Leu Cys Leu Leu Gly Arg Lys Tyr Ile Leu Ala Pro Ala 20 25 30 His His Val Glu Ser Ala 35
Claims (7)
Porcine reproductive and respiratory syndrome virus immunogenic peptide consisting of the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2.
An immunogenic composition against porcine reproductive and respiratory syndrome virus comprising the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2.
상기 면역원성 조성물은,
백신 또는 백신 조성물인 것을 특징으로 하는 면역원성 조성물.
According to paragraph 2,
The immunogenic composition,
An immunogenic composition, characterized in that it is a vaccine or vaccine composition.
상기 면역원성 조성물은,
T 세포에서 인터페론-감마 (INF-γ)의 발현을 유도하는 것을 특징으로 하는 면역원성 조성물.
According to paragraph 2,
The immunogenic composition,
An immunogenic composition characterized in that it induces the expression of interferon-gamma (INF-γ) in T cells.
A pharmaceutical composition for preventing or treating porcine reproductive and respiratory syndrome, comprising the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2.
A feed composition for preventing or improving porcine reproductive and respiratory syndrome, comprising the peptide of SEQ ID NO: 1 or the peptide of SEQ ID NO: 2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020210046000A KR102604839B1 (en) | 2021-04-08 | 2021-04-08 | Composition for antigenicity comprising peptide from PRRSV |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020210046000A KR102604839B1 (en) | 2021-04-08 | 2021-04-08 | Composition for antigenicity comprising peptide from PRRSV |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| KR20220139705A KR20220139705A (en) | 2022-10-17 |
| KR102604839B1 true KR102604839B1 (en) | 2023-11-22 |
Family
ID=83809935
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020210046000A KR102604839B1 (en) | 2021-04-08 | 2021-04-08 | Composition for antigenicity comprising peptide from PRRSV |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR102604839B1 (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102193460B1 (en) | 2020-03-24 | 2020-12-21 | 주식회사 바이오포아 | Vaccine composition for preventing porcine reproductive and respiratory syndrome and preventing method thereof |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7829274B2 (en) * | 2007-09-04 | 2010-11-09 | Boehringer Ingelheim Vetmedica, Inc. | Reduction of concomitant infections in pigs by the use of PCV2 antigen |
| KR101933240B1 (en) * | 2014-06-12 | 2018-12-28 | 건국대학교 산학협력단 | Differentiating Infected from Vaccinated Animals Vacine for Porcine Reproductive and Respiratory Syndrome Virus |
| KR101933932B1 (en) | 2017-12-27 | 2018-12-31 | 주식회사 바이오포아 | Mutant strain of porcine reproductive and respiratory syndrome virus and antiviral vaccines including the same |
| KR102481135B1 (en) * | 2018-01-02 | 2022-12-26 | 한국생명공학연구원 | Composition for preventing or treating infectious disease containing extract of Celastrus orbiculatus Thunb |
-
2021
- 2021-04-08 KR KR1020210046000A patent/KR102604839B1/en active IP Right Grant
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102193460B1 (en) | 2020-03-24 | 2020-12-21 | 주식회사 바이오포아 | Vaccine composition for preventing porcine reproductive and respiratory syndrome and preventing method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| KR20220139705A (en) | 2022-10-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR20200132931A (en) | Antiviral immunosuppressant for the treatment of acute respiratory viral infections | |
| CN113454102A (en) | African swine fever vaccine | |
| AU2018278927A1 (en) | Methods and compositions for dengue virus vaccines | |
| KR20170075775A (en) | Methods and compositions for recombinant dengue viruses for vaccine and diagnostic development | |
| KR20110090711A (en) | Novel porcine circovirus type 2 and use thereof | |
| JP7046607B2 (en) | Pestivirus vaccine against congenital tremor | |
| KR102136954B1 (en) | A composition for preventing or treating MERS-CoV virus comprising HBD2 (human beta-defensin 2) or an fusion protein comprising epitope protein of MERS-CoV virus and the HBD2) | |
| KR20190070629A (en) | A composition for preventing or treating MERS-CoV virus comprising HBD2 (human beta-defensin 2) or an fusion protein comprising epitope protein of MERS-CoV virus and the HBD2L | |
| KR102604839B1 (en) | Composition for antigenicity comprising peptide from PRRSV | |
| JPH07504662A (en) | Immune stimulants for therapeutic use in immunocompromised hosts | |
| KR20170055766A (en) | Vaccine composition for preventing or treating brucellosis comprising attenuated Salmonella mutant as effective component | |
| KR102062624B1 (en) | Vaccine composition for preventing or treating avian influenza and fowl typhoid simultaneouly comprising attenuated Salmonella gallinarum mutant as effective component | |
| KR101579861B1 (en) | Vaccine composition for tuberculosis containing Rv3131 protein | |
| KR20220133633A (en) | Recombinant protein comprising spike protein S1 of SARS-CoV-2-derived protein and ferritin-derived protein and use thereof | |
| KR101642705B1 (en) | Attenuated genotype 1 Japanese encephalitis vaccine composition containing the same for preventing Japanese encephalitis in swine | |
| KR20220133631A (en) | Recombinant protein comprising spike protein of SARS-CoV-2-derived protein and Fc of immunoglobulin-derived protein and use thereof | |
| CN116133669A (en) | Materials and methods for inhibiting viral infections, including coronavirus infections | |
| KR101705769B1 (en) | Vaccine composition for tuberculosis containing MTBK_20640 protein | |
| KR20150101630A (en) | Vaccine composition for tuberculosis containing Rv0351 protein | |
| KR102221244B1 (en) | Vaccine composition for preventing tuberculosis comprising Mycobacterium anyangense strain | |
| KR102616625B1 (en) | Vaccine composition for preventing or treating of caseous lymphadenitis and uses thereof | |
| KR20140064847A (en) | Use of the pacap as a molecular adjuvant for vaccines | |
| KR102336245B1 (en) | Vaccine composition of novel inactivated Actinobacillus pleuroneumoniae bacteria | |
| CN108419437A (en) | DNA vaccination for preventing and treating HSV-2 infection | |
| KR101825090B1 (en) | Pharmaceutical Composition Comprising Ribavirin Or Derivatives Thereof For Treating Foot-And-Mouth Disease |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| E701 | Decision to grant or registration of patent right |