WO2022260468A1 - 신규 fgf21 변이체 개발 및 이의 생산기법과 용도 - Google Patents
신규 fgf21 변이체 개발 및 이의 생산기법과 용도 Download PDFInfo
- Publication number
- WO2022260468A1 WO2022260468A1 PCT/KR2022/008202 KR2022008202W WO2022260468A1 WO 2022260468 A1 WO2022260468 A1 WO 2022260468A1 KR 2022008202 W KR2022008202 W KR 2022008202W WO 2022260468 A1 WO2022260468 A1 WO 2022260468A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- fgf21
- protein
- variant
- mutant protein
- polypeptide
- Prior art date
Links
- 102000003973 Fibroblast growth factor 21 Human genes 0.000 title claims abstract description 146
- 108090000376 Fibroblast growth factor 21 Proteins 0.000 title claims abstract description 146
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 17
- 238000000034 method Methods 0.000 title claims description 37
- 238000011161 development Methods 0.000 title abstract description 3
- 230000000694 effects Effects 0.000 claims abstract description 22
- 108090000623 proteins and genes Proteins 0.000 claims description 56
- 102000004169 proteins and genes Human genes 0.000 claims description 44
- 108010021466 Mutant Proteins Proteins 0.000 claims description 43
- 102000008300 Mutant Proteins Human genes 0.000 claims description 43
- 150000001413 amino acids Chemical class 0.000 claims description 28
- 238000010438 heat treatment Methods 0.000 claims description 27
- 239000000203 mixture Substances 0.000 claims description 17
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 17
- 239000008194 pharmaceutical composition Substances 0.000 claims description 15
- 208000030159 metabolic disease Diseases 0.000 claims description 14
- 229920001184 polypeptide Polymers 0.000 claims description 14
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 14
- 208000024172 Cardiovascular disease Diseases 0.000 claims description 13
- 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 7
- 230000035772 mutation Effects 0.000 claims description 7
- 238000006467 substitution reaction Methods 0.000 claims description 7
- 239000001963 growth medium Substances 0.000 claims description 6
- 102220198580 rs113903766 Human genes 0.000 claims description 6
- 102200076716 rs374059924 Human genes 0.000 claims description 6
- 102220222820 rs759405317 Human genes 0.000 claims description 6
- 108010076504 Protein Sorting Signals Proteins 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 5
- 239000012535 impurity Substances 0.000 claims description 5
- 238000000746 purification Methods 0.000 claims description 5
- 102220577033 Ornithine decarboxylase_Q184E_mutation Human genes 0.000 claims description 3
- 125000003275 alpha amino acid group Chemical group 0.000 claims 2
- 238000007792 addition Methods 0.000 claims 1
- 230000037430 deletion Effects 0.000 claims 1
- 238000012217 deletion Methods 0.000 claims 1
- 230000001965 increasing effect Effects 0.000 abstract description 5
- 229940000406 drug candidate Drugs 0.000 abstract description 3
- 230000019491 signal transduction Effects 0.000 abstract description 3
- 235000018102 proteins Nutrition 0.000 description 39
- 235000001014 amino acid Nutrition 0.000 description 21
- 229940024606 amino acid Drugs 0.000 description 20
- 210000004027 cell Anatomy 0.000 description 13
- 102100020683 Beta-klotho Human genes 0.000 description 12
- 101710104526 Beta-klotho Proteins 0.000 description 12
- 229920002971 Heparan sulfate Polymers 0.000 description 10
- 230000006870 function Effects 0.000 description 9
- 208000008338 non-alcoholic fatty liver disease Diseases 0.000 description 9
- 239000013598 vector Substances 0.000 description 9
- 108091008794 FGF receptors Proteins 0.000 description 8
- 230000004913 activation Effects 0.000 description 8
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Natural products NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 7
- 108091005804 Peptidases Proteins 0.000 description 7
- 239000004365 Protease Substances 0.000 description 7
- 239000000796 flavoring agent Substances 0.000 description 7
- 206010053219 non-alcoholic steatohepatitis Diseases 0.000 description 7
- 239000004471 Glycine Substances 0.000 description 6
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 6
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- 108060008226 thioredoxin Proteins 0.000 description 6
- 102000018233 Fibroblast Growth Factor Human genes 0.000 description 5
- 108050007372 Fibroblast Growth Factor Proteins 0.000 description 5
- 102000044168 Fibroblast Growth Factor Receptor Human genes 0.000 description 5
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 5
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 5
- -1 Polyethylen Polymers 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 238000005119 centrifugation Methods 0.000 description 5
- 238000012136 culture method Methods 0.000 description 5
- 229940126864 fibroblast growth factor Drugs 0.000 description 5
- 230000036541 health Effects 0.000 description 5
- 229960002897 heparin Drugs 0.000 description 5
- 239000000546 pharmaceutical excipient Substances 0.000 description 5
- 230000001766 physiological effect Effects 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 102000005962 receptors Human genes 0.000 description 5
- 108020003175 receptors Proteins 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 4
- 102100036407 Thioredoxin Human genes 0.000 description 4
- 241000723792 Tobacco etch virus Species 0.000 description 4
- 150000001720 carbohydrates Chemical class 0.000 description 4
- 235000014633 carbohydrates Nutrition 0.000 description 4
- 238000004925 denaturation Methods 0.000 description 4
- 230000036425 denaturation Effects 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 239000000839 emulsion Substances 0.000 description 4
- 235000019634 flavors Nutrition 0.000 description 4
- 238000009472 formulation Methods 0.000 description 4
- 235000013376 functional food Nutrition 0.000 description 4
- 210000004185 liver Anatomy 0.000 description 4
- 108091033319 polynucleotide Proteins 0.000 description 4
- 102000040430 polynucleotide Human genes 0.000 description 4
- 239000002157 polynucleotide Substances 0.000 description 4
- 150000003839 salts Chemical class 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 230000014616 translation Effects 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 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 3
- 241000701489 Cauliflower mosaic virus Species 0.000 description 3
- 208000032928 Dyslipidaemia Diseases 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- 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 3
- 208000035150 Hypercholesterolemia Diseases 0.000 description 3
- 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 3
- 208000017170 Lipid metabolism disease Diseases 0.000 description 3
- 102000035195 Peptidases Human genes 0.000 description 3
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 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 3
- 229930006000 Sucrose Natural products 0.000 description 3
- 239000004480 active ingredient Substances 0.000 description 3
- 210000001789 adipocyte Anatomy 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
- 238000012790 confirmation Methods 0.000 description 3
- 239000013078 crystal Substances 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 102000052178 fibroblast growth factor receptor activity proteins Human genes 0.000 description 3
- 235000013355 food flavoring agent Nutrition 0.000 description 3
- 230000002068 genetic effect Effects 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 125000003630 glycyl group Chemical group [H]N([H])C([H])([H])C(*)=O 0.000 description 3
- 208000006575 hypertriglyceridemia Diseases 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 239000008101 lactose Substances 0.000 description 3
- 210000000496 pancreas Anatomy 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- 235000000346 sugar Nutrition 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 239000000829 suppository Substances 0.000 description 3
- 101150003509 tag gene Proteins 0.000 description 3
- LWTDZKXXJRRKDG-KXBFYZLASA-N (-)-phaseollin Chemical compound C1OC2=CC(O)=CC=C2[C@H]2[C@@H]1C1=CC=C3OC(C)(C)C=CC3=C1O2 LWTDZKXXJRRKDG-KXBFYZLASA-N 0.000 description 2
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 2
- 241000589155 Agrobacterium tumefaciens Species 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 241000701022 Cytomegalovirus Species 0.000 description 2
- IMXSCCDUAFEIOE-UHFFFAOYSA-N D-Octopin Natural products OC(=O)C(C)NC(C(O)=O)CCCN=C(N)N IMXSCCDUAFEIOE-UHFFFAOYSA-N 0.000 description 2
- IMXSCCDUAFEIOE-RITPCOANSA-N D-octopine Chemical compound [O-]C(=O)[C@@H](C)[NH2+][C@H](C([O-])=O)CCCNC(N)=[NH2+] IMXSCCDUAFEIOE-RITPCOANSA-N 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- YQYJSBFKSSDGFO-UHFFFAOYSA-N Epihygromycin Natural products OC1C(O)C(C(=O)C)OC1OC(C(=C1)O)=CC=C1C=C(C)C(=O)NC1C(O)C(O)C2OCOC2C1O YQYJSBFKSSDGFO-UHFFFAOYSA-N 0.000 description 2
- 101150021185 FGF gene Proteins 0.000 description 2
- 108010074860 Factor Xa Proteins 0.000 description 2
- 229930091371 Fructose Natural products 0.000 description 2
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 2
- 239000005715 Fructose Substances 0.000 description 2
- 229920002683 Glycosaminoglycan Polymers 0.000 description 2
- AVXURJPOCDRRFD-UHFFFAOYSA-N Hydroxylamine Chemical compound ON AVXURJPOCDRRFD-UHFFFAOYSA-N 0.000 description 2
- 241000829100 Macaca mulatta polyomavirus 1 Species 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 208000008589 Obesity Diseases 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 102000004022 Protein-Tyrosine Kinases Human genes 0.000 description 2
- 108090000412 Protein-Tyrosine Kinases Proteins 0.000 description 2
- 241000725643 Respiratory syncytial virus Species 0.000 description 2
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 244000299461 Theobroma cacao Species 0.000 description 2
- 102000002933 Thioredoxin Human genes 0.000 description 2
- 108090000190 Thrombin Proteins 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 210000000577 adipose tissue Anatomy 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 239000003125 aqueous solvent Substances 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 210000004556 brain Anatomy 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 230000002124 endocrine Effects 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 230000004153 glucose metabolism Effects 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 238000007912 intraperitoneal administration Methods 0.000 description 2
- 230000037356 lipid metabolism Effects 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
- 239000003550 marker Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 108010058731 nopaline synthase Proteins 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 235000020824 obesity Nutrition 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 235000005985 organic acids Nutrition 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 229910052698 phosphorus Inorganic materials 0.000 description 2
- 239000006187 pill Substances 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 150000004804 polysaccharides Chemical class 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- HELXLJCILKEWJH-NCGAPWICSA-N rebaudioside A Chemical compound O([C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O HELXLJCILKEWJH-NCGAPWICSA-N 0.000 description 2
- 238000003259 recombinant expression Methods 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 230000011664 signaling Effects 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000000527 sonication Methods 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
- 239000006188 syrup Substances 0.000 description 2
- 235000020357 syrup Nutrition 0.000 description 2
- 229940124597 therapeutic agent Drugs 0.000 description 2
- 229940094937 thioredoxin Drugs 0.000 description 2
- 229960004072 thrombin Drugs 0.000 description 2
- 238000013518 transcription Methods 0.000 description 2
- 230000035897 transcription Effects 0.000 description 2
- 238000011426 transformation method Methods 0.000 description 2
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 239000000080 wetting agent Substances 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 description 1
- 108020003589 5' Untranslated Regions Proteins 0.000 description 1
- 102000007469 Actins Human genes 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 241000589158 Agrobacterium Species 0.000 description 1
- 108010011170 Ala-Trp-Arg-His-Pro-Gln-Phe-Gly-Gly Proteins 0.000 description 1
- ATRRKUHOCOJYRX-UHFFFAOYSA-N Ammonium bicarbonate Chemical compound [NH4+].OC([O-])=O ATRRKUHOCOJYRX-UHFFFAOYSA-N 0.000 description 1
- 239000004254 Ammonium phosphate Substances 0.000 description 1
- 239000004382 Amylase Substances 0.000 description 1
- 102000013142 Amylases Human genes 0.000 description 1
- 108010065511 Amylases Proteins 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- 108010011485 Aspartame Proteins 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000283254 Balaenoptera acutorostrata Species 0.000 description 1
- 108010006654 Bleomycin Proteins 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 102000000584 Calmodulin Human genes 0.000 description 1
- 108010041952 Calmodulin Proteins 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- ZEOWTGPWHLSLOG-UHFFFAOYSA-N Cc1ccc(cc1-c1ccc2c(n[nH]c2c1)-c1cnn(c1)C1CC1)C(=O)Nc1cccc(c1)C(F)(F)F Chemical compound Cc1ccc(cc1-c1ccc2c(n[nH]c2c1)-c1cnn(c1)C1CC1)C(=O)Nc1cccc(c1)C(F)(F)F ZEOWTGPWHLSLOG-UHFFFAOYSA-N 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 229920000858 Cyclodextrin Polymers 0.000 description 1
- 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 1
- 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 1
- 230000004543 DNA replication Effects 0.000 description 1
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 1
- 108090000626 DNA-directed RNA polymerases Proteins 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 241000283086 Equidae Species 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
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 239000001512 FEMA 4601 Substances 0.000 description 1
- 102000003971 Fibroblast Growth Factor 1 Human genes 0.000 description 1
- 108090000386 Fibroblast Growth Factor 1 Proteins 0.000 description 1
- 102100024802 Fibroblast growth factor 23 Human genes 0.000 description 1
- 102100028072 Fibroblast growth factor 4 Human genes 0.000 description 1
- 102100028071 Fibroblast growth factor 7 Human genes 0.000 description 1
- 102100037680 Fibroblast growth factor 8 Human genes 0.000 description 1
- 102100037665 Fibroblast growth factor 9 Human genes 0.000 description 1
- 102100023593 Fibroblast growth factor receptor 1 Human genes 0.000 description 1
- 101710182386 Fibroblast growth factor receptor 1 Proteins 0.000 description 1
- 239000004606 Fillers/Extenders Substances 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 239000004378 Glycyrrhizin Substances 0.000 description 1
- 239000005562 Glyphosate Substances 0.000 description 1
- HVLSXIKZNLPZJJ-TXZCQADKSA-N HA peptide Chemical compound C([C@@H](C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](N)CC=1C=CC(O)=CC=1)C1=CC=C(O)C=C1 HVLSXIKZNLPZJJ-TXZCQADKSA-N 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 108010033040 Histones Proteins 0.000 description 1
- 101000812437 Homo sapiens ER lumen protein-retaining receptor 1 Proteins 0.000 description 1
- 101100120063 Homo sapiens FGF21 gene Proteins 0.000 description 1
- 101001051973 Homo sapiens Fibroblast growth factor 23 Proteins 0.000 description 1
- 101001060274 Homo sapiens Fibroblast growth factor 4 Proteins 0.000 description 1
- 101001060261 Homo sapiens Fibroblast growth factor 7 Proteins 0.000 description 1
- 101001027382 Homo sapiens Fibroblast growth factor 8 Proteins 0.000 description 1
- 101001027380 Homo sapiens Fibroblast growth factor 9 Proteins 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
- 102000015834 Klotho Human genes 0.000 description 1
- 108050004036 Klotho Proteins 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 1
- 108010028554 LDL Cholesterol Proteins 0.000 description 1
- OYHQOLUKZRVURQ-HZJYTTRNSA-N Linoleic acid Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(O)=O OYHQOLUKZRVURQ-HZJYTTRNSA-N 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 101100390675 Mus musculus Fgf15 gene Proteins 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 235000021314 Palmitic acid Nutrition 0.000 description 1
- 235000019483 Peanut oil Nutrition 0.000 description 1
- 229920002230 Pectic acid Polymers 0.000 description 1
- 102000010292 Peptide Elongation Factor 1 Human genes 0.000 description 1
- 108010077524 Peptide Elongation Factor 1 Proteins 0.000 description 1
- 102000005877 Peptide Initiation Factors Human genes 0.000 description 1
- 108010044843 Peptide Initiation Factors Proteins 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 101710163504 Phaseolin Proteins 0.000 description 1
- IAJOBQBIJHVGMQ-UHFFFAOYSA-N Phosphinothricin Natural products CP(O)(=O)CCC(N)C(O)=O IAJOBQBIJHVGMQ-UHFFFAOYSA-N 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 108010020346 Polyglutamic Acid Proteins 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- HELXLJCILKEWJH-SEAGSNCFSA-N Rebaudioside A Natural products O=C(O[C@H]1[C@@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1)[C@@]1(C)[C@@H]2[C@](C)([C@H]3[C@@]4(CC(=C)[C@@](O[C@H]5[C@H](O[C@H]6[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O6)[C@@H](O[C@H]6[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O6)[C@H](O)[C@@H](CO)O5)(C4)CC3)CC2)CCC1 HELXLJCILKEWJH-SEAGSNCFSA-N 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 235000019764 Soybean Meal Nutrition 0.000 description 1
- 235000021355 Stearic acid Nutrition 0.000 description 1
- 244000228451 Stevia rebaudiana Species 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 235000019486 Sunflower oil Nutrition 0.000 description 1
- 235000005764 Theobroma cacao ssp. cacao Nutrition 0.000 description 1
- 235000005767 Theobroma cacao ssp. sphaerocarpum Nutrition 0.000 description 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- 108090000848 Ubiquitin Proteins 0.000 description 1
- 102000044159 Ubiquitin Human genes 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 101150067314 aadA gene Proteins 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 239000000556 agonist Substances 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 239000001099 ammonium carbonate Substances 0.000 description 1
- 235000012501 ammonium carbonate Nutrition 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 229910000148 ammonium phosphate Inorganic materials 0.000 description 1
- 235000019289 ammonium phosphates Nutrition 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 235000019418 amylase Nutrition 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- 239000000605 aspartame Substances 0.000 description 1
- 235000010357 aspartame Nutrition 0.000 description 1
- IAOZJIPTCAWIRG-QWRGUYRKSA-N aspartame Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(=O)OC)CC1=CC=CC=C1 IAOZJIPTCAWIRG-QWRGUYRKSA-N 0.000 description 1
- 229960003438 aspartame Drugs 0.000 description 1
- 150000007514 bases Chemical class 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 229960001561 bleomycin Drugs 0.000 description 1
- OYVAGSVQBOHSSS-UAPAGMARSA-O bleomycin A2 Chemical compound N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC=C(N=1)C=1SC=C(N=1)C(=O)NCCC[S+](C)C)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1N=CNC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C OYVAGSVQBOHSSS-UAPAGMARSA-O 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 235000014121 butter Nutrition 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 235000001046 cacaotero Nutrition 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 235000014171 carbonated beverage Nutrition 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 235000013351 cheese Nutrition 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 1
- 229960005091 chloramphenicol Drugs 0.000 description 1
- 235000019219 chocolate Nutrition 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 231100000313 clinical toxicology Toxicity 0.000 description 1
- 239000003240 coconut oil Substances 0.000 description 1
- 235000019864 coconut oil Nutrition 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 229940097362 cyclodextrins Drugs 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 229940125542 dual agonist Drugs 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 210000002472 endoplasmic reticulum Anatomy 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- HELXLJCILKEWJH-UHFFFAOYSA-N entered according to Sigma 01432 Natural products C1CC2C3(C)CCCC(C)(C(=O)OC4C(C(O)C(O)C(CO)O4)O)C3CCC2(C2)CC(=C)C21OC(C1OC2C(C(O)C(O)C(CO)O2)O)OC(CO)C(O)C1OC1OC(CO)C(O)C(O)C1O HELXLJCILKEWJH-UHFFFAOYSA-N 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 235000019414 erythritol Nutrition 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
- 229940009714 erythritol Drugs 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000003925 fat Substances 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 208000010706 fatty liver disease Diseases 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 239000005417 food ingredient Substances 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 229920000370 gamma-poly(glutamate) polymer Polymers 0.000 description 1
- 239000007789 gas 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
- BRZYSWJRSDMWLG-CAXSIQPQSA-N geneticin Natural products O1C[C@@](O)(C)[C@H](NC)[C@@H](O)[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](C(C)O)O2)N)[C@@H](N)C[C@H]1N BRZYSWJRSDMWLG-CAXSIQPQSA-N 0.000 description 1
- IAJOBQBIJHVGMQ-BYPYZUCNSA-N glufosinate-P Chemical compound CP(O)(=O)CC[C@H](N)C(O)=O IAJOBQBIJHVGMQ-BYPYZUCNSA-N 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- LPLVUJXQOOQHMX-UHFFFAOYSA-N glycyrrhetinic acid glycoside Natural products C1CC(C2C(C3(CCC4(C)CCC(C)(CC4C3=CC2=O)C(O)=O)C)(C)CC2)(C)C2C(C)(C)C1OC1OC(C(O)=O)C(O)C(O)C1OC1OC(C(O)=O)C(O)C(O)C1O LPLVUJXQOOQHMX-UHFFFAOYSA-N 0.000 description 1
- 229960004949 glycyrrhizic acid Drugs 0.000 description 1
- UYRUBYNTXSDKQT-UHFFFAOYSA-N glycyrrhizic acid Natural products CC1(C)C(CCC2(C)C1CCC3(C)C2C(=O)C=C4C5CC(C)(CCC5(C)CCC34C)C(=O)O)OC6OC(C(O)C(O)C6OC7OC(O)C(O)C(O)C7C(=O)O)C(=O)O UYRUBYNTXSDKQT-UHFFFAOYSA-N 0.000 description 1
- 235000019410 glycyrrhizin Nutrition 0.000 description 1
- LPLVUJXQOOQHMX-QWBHMCJMSA-N glycyrrhizinic acid Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@@H]1C([C@H]2[C@]([C@@H]3[C@@]([C@@]4(CC[C@@]5(C)CC[C@@](C)(C[C@H]5C4=CC3=O)C(O)=O)C)(C)CC2)(C)CC1)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O LPLVUJXQOOQHMX-QWBHMCJMSA-N 0.000 description 1
- XDDAORKBJWWYJS-UHFFFAOYSA-N glyphosate Chemical compound OC(=O)CNCP(O)(O)=O XDDAORKBJWWYJS-UHFFFAOYSA-N 0.000 description 1
- 229940097068 glyphosate Drugs 0.000 description 1
- 230000002363 herbicidal effect Effects 0.000 description 1
- 239000004009 herbicide Substances 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 125000000487 histidyl group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 150000002484 inorganic compounds Chemical class 0.000 description 1
- 229910010272 inorganic material Inorganic materials 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000001361 intraarterial administration Methods 0.000 description 1
- 230000031146 intracellular signal transduction Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 229910000358 iron sulfate Inorganic materials 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229930027917 kanamycin Natural products 0.000 description 1
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 1
- 229960000318 kanamycin Drugs 0.000 description 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- VMPHSYLJUKZBJJ-UHFFFAOYSA-N lauric acid triglyceride Natural products CCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCC)COC(=O)CCCCCCCCCCC VMPHSYLJUKZBJJ-UHFFFAOYSA-N 0.000 description 1
- 235000020778 linoleic acid Nutrition 0.000 description 1
- OYHQOLUKZRVURQ-IXWMQOLASA-N linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 229940057995 liquid paraffin Drugs 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 229960003511 macrogol Drugs 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate 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
- 108020004999 messenger RNA Proteins 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 239000012457 nonaqueous media Substances 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 235000008390 olive oil Nutrition 0.000 description 1
- 230000003076 paracrine Effects 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000000312 peanut oil Substances 0.000 description 1
- LCLHHZYHLXDRQG-ZNKJPWOQSA-N pectic acid Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)O[C@H](C(O)=O)[C@@H]1OC1[C@H](O)[C@@H](O)[C@@H](OC2[C@@H]([C@@H](O)[C@@H](O)[C@H](O2)C(O)=O)O)[C@@H](C(O)=O)O1 LCLHHZYHLXDRQG-ZNKJPWOQSA-N 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- LWTDZKXXJRRKDG-UHFFFAOYSA-N phaseollin Natural products C1OC2=CC(O)=CC=C2C2C1C1=CC=C3OC(C)(C)C=CC3=C1O2 LWTDZKXXJRRKDG-UHFFFAOYSA-N 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 239000010318 polygalacturonic acid Substances 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- GCYXWQUSHADNBF-AAEALURTSA-N preproglucagon 78-108 Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1N=CNC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=CC=C1 GCYXWQUSHADNBF-AAEALURTSA-N 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000012846 protein folding Effects 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 235000019203 rebaudioside A Nutrition 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 235000019204 saccharin Nutrition 0.000 description 1
- CVHZOJJKTDOEJC-UHFFFAOYSA-N saccharin Chemical compound C1=CC=C2C(=O)NS(=O)(=O)C2=C1 CVHZOJJKTDOEJC-UHFFFAOYSA-N 0.000 description 1
- 229940081974 saccharin Drugs 0.000 description 1
- 239000000901 saccharin and its Na,K and Ca salt Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000007781 signaling event Effects 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 229960002920 sorbitol Drugs 0.000 description 1
- 235000010356 sorbitol Nutrition 0.000 description 1
- 239000004455 soybean meal Substances 0.000 description 1
- 239000003549 soybean oil Substances 0.000 description 1
- 235000012424 soybean oil Nutrition 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- 238000012916 structural analysis Methods 0.000 description 1
- 150000005846 sugar alcohols Chemical class 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L sulfate group Chemical group S(=O)(=O)([O-])[O-] QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 239000002600 sunflower oil Substances 0.000 description 1
- 239000010414 supernatant solution Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000002195 synergetic effect Effects 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
- 238000012360 testing method Methods 0.000 description 1
- 239000000892 thaumatin Substances 0.000 description 1
- 235000010436 thaumatin Nutrition 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 125000000341 threoninyl group Chemical group [H]OC([H])(C([H])([H])[H])C([H])(N([H])[H])C(*)=O 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000005026 transcription initiation Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 150000003626 triacylglycerols Chemical class 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000001262 western blot Methods 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
- 239000012138 yeast extract Substances 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/475—Growth factors; Growth regulators
- C07K14/50—Fibroblast growth factors [FGF]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/35—Fusion polypeptide containing a fusion for enhanced stability/folding during expression, e.g. fusions with chaperones or thioredoxin
Definitions
- FGF Fibroblast growth factor
- FGFs belonging to five subfamilies require heparin or heparan sulfate when binding to receptors.
- Heparin/heparan sulfate present on the cell surface or extracellular matrix is a negatively charged polysaccharide belonging to the Glycosaminoglycan (GAG) family and interacts with various proteins to regulate their functions.
- GAG Glycosaminoglycan
- FGF21 is a protein encoded by the FGF21 gene in mammals and belongs to the endocrine subfamily including FGF23 and FGF15/19.
- FGF21 is the major endogenous agonist of the FGF21 receptor, which consists of co-receptors FGF receptor 1c and ⁇ -Klotho.
- FGF21 has recently been widely studied as an active ingredient in various therapeutic agents. Specifically, in July 2019, Yuhan Corporation signed a contract with Boehringer, a German multinational pharmaceutical company, for the purpose of developing a treatment for nonalcoholic steatohepatitis (NASH).
- NASH nonalcoholic steatohepatitis
- the present inventors have made intensive efforts to develop a FGF21 protein variant with improved activity, and as a result, developed a mutant with about 6-fold increase in signaling activity compared to wild type, and completed the present invention.
- One aspect relates to novel FGF21 variant proteins and methods for their production.
- One aspect provides a polypeptide comprising a FGF21 variant, wherein the variant contains one or more mutations in the wild-type FGF21 protein.
- Another aspect is culturing a transformant producing a FGF21 mutant protein; And it provides a method for producing a FGF21 mutant protein comprising the step of heating the transformant or culture medium to a temperature of 40 ° C. or higher.
- Another aspect provides a method for purifying a FGF21 mutant protein, comprising heating a mixture containing the FGF21 mutant protein and impurities to a temperature of 40° C. or higher.
- Another aspect provides a FGF21 mutant protein prepared by the above method.
- Another aspect provides a pharmaceutical composition for treating metabolic disorders or cardiovascular disorders containing the FGF21 mutant protein.
- Another aspect provides a method for treating metabolic disorders or cardiovascular disorders, comprising administering the pharmaceutical composition to a subject.
- Another aspect provides the use of the pharmaceutical composition for the treatment of metabolic disorders or cardiovascular disorders.
- the novel FGF21 mutant since the novel FGF21 mutant according to one embodiment has a signal transduction activity increased by about 6 times compared to the wild type, the mutant can be evaluated as a valuable drug candidate as a biobetter.
- 1 is a diagram showing the results of sequence comparison and analysis of hFGF21 and wFGF21, and shaded marks indicate sequences with specific differences among the C-terminal tails of the two sequences.
- FIG. 2 is a view showing the results of an X-ray crystal structure (PDB code: 5VAQ) analysis of the complex of hFGF21CT and ⁇ -Klotho, and the sequences corresponding to variant design strategies 1 and 2 in the hFGF21CT structure are enlarged and displayed.
- PDB code 5VAQ
- Figure 3 is a diagram showing the FGF21 wild-type sequence and mutant sequence.
- FIG. 4 is a view showing the SDS-PAGE analysis results (left) of the supernatant in which the Trx-FGF21 mutant is overexpressed before and after heat treatment and the SDS-PAGE analysis result (right) of the final purified FGF21 mutant 3.
- FIG. 5 is a diagram showing data comparing the activities of commercially available FGF21 wild-type and heat-treated purified FGF21 N4.
- FIG. 6 is a diagram showing in vitro cell assay data of FGF21 wild-type and mutants 1, 2, and 3 for 3T3L1 adipocytes.
- FIG. 7 is a view showing FGFR1c activation experimental data according to concentrations of FGF21 wild-type and variants 1, 2 and 3.
- the term “combination(s) of these” included in the expression of the Markush form means a mixture or combination of one or more selected from the group consisting of the components described in the expression of the Markush form, It means including one or more selected from the group consisting of components.
- One aspect provides a polypeptide comprising a FGF21 variant, wherein the variant contains one or more mutations in the wild-type FGF21 protein.
- fibroblast growth factor 21 used throughout this specification is a protein consisting of 209 amino acids
- FGF21 functions as an endocrine hormone that reaches distant targets and exerts physiological activity.
- FGF21 acts as a complex with FGFR1c, one of the FGF receptors present on the cell surface, and the cofactor ⁇ -Klotho protein.
- FGFR1c one of the FGF receptors present on the cell surface
- ⁇ -Klotho the cofactor of ⁇ -Klotho protein.
- Heparin/heparan sulfate is present in all cells, whereas ⁇ -Klotho is mainly distributed in adipose tissue, liver, and pancreas.
- FGF21 regulates glucose and lipid metabolism by influencing the activity of cells expressing FGFR1c and ⁇ -Klotho.
- FGF21 has been reported to be effective in improving metabolic diseases such as type 2 diabetes and fatty liver disease. has emerged as an important candidate material.
- FGF21 variant refers to a variant comprising a mutation in which one or more amino acids are substituted, deleted, or added in the amino acid sequence of wild type FGF21.
- FGF21 variant may be used interchangeably with “FGF variant protein”.
- the polypeptide may include a FGF21 variant, and specifically may be composed of the FGF21 variant.
- the wild-type FGF21 protein may be in a mature form in which the signal peptide is removed, and specifically, in the FGF21 protein composed of SEQ ID NO: 1, amino acids 1 to 28, which are signal peptides, are removed to form the 29th amino acid. to 209th amino acid, and more specifically, it may include the amino acid of SEQ ID NO: 2.
- the FGF21 mutant may be one in which 1 to 4 amino acids at the N-terminus are deleted.
- the FGF21 variant may be one in which one or more amino acids selected from the group consisting of Q184, S195, P199, S200, Q201 and A208 are substituted with other amino acids, specifically 20 amino acids (Q, E, G , A, S, T, C, V, L, I, M, P, F, Y, W, D, N, H, K, R) may be substituted with one of.
- the FGF21 variant may be substituted with one or more selected from the group consisting of Q184E, S195G, P199G, S200G, Q201H and A208T, specifically S195G, P199G, S200G, Q201H and A208T.
- the S195G substitution is that the 167th amino acid (serine) of SEQ ID NO: 2 is substituted with glycine
- the P199G substitution is that the 171st amino acid (proline) of SEQ ID NO: 2 is substituted with glycine
- the S200G substitution is the sequence
- the 172nd amino acid (serine) of SEQ ID NO: 2 is substituted with glycine
- the Q201H substitution is that the 173rd amino acid (glutamine) of SEQ ID NO: 2 is substituted with histidine
- the A208T substitution is that the 180th amino acid (alanine) of SEQ ID NO: 2 is substituted. It may be substituted with threonine.
- the FGF21 variant may be one of the amino acid sequences of SEQ ID NOs: 3 to 6.
- the FGF21 variant may have improved activity compared to the wild type, and specifically, signal transduction activity may be improved by about 6 times compared to the wild type.
- Another aspect is culturing a transformant producing a FGF21 mutant protein; And it provides a method for producing a FGF21 mutant protein comprising the step of heating the transformant or culture medium to a temperature of 40 ° C. or higher.
- the contents overlapping with the FGF21 mutant-containing polypeptide are also applied to the production method of the FGF21 mutant protein.
- the FGF21 mutant protein may include a polypeptide comprising at least one of the amino acid sequences of SEQ ID NOs: 3 to 6, and may specifically include the amino acid sequence of SEQ ID NO: 6.
- the transformant may include a polynucleotide encoding the FGF21 mutant protein, and specifically, a gene construct or a recombinant vector including a polynucleotide encoding the FGF21 mutant protein.
- the transformant producing the FGF21 mutant protein may express the FGF21 mutant by including a gene construct or a recombinant vector containing a polynucleotide encoding the FGF21 protein.
- the term "recombinant vector” refers to a vector capable of expressing a peptide or protein encoded by a heterologous nucleic acid inserted into the vector, preferably a polynucleotide encoding the FGF21 protein. Refers to a vector prepared to contain a nucleotide.
- the "vector” refers to any medium for the introduction and/or transfer of a base into a host cell in vitro, in vivo, or in vivo, and other DNA fragments bind to and bind to each other.
- a replica unit is any genetic unit that functions as a self-unit of DNA replication in vivo, that is, can be replicated by its own regulation ( For example, plasmid, phage, cosmid, chromosome, virus, etc.).
- the recombinant vector or recombinant expression vector preferably includes a promoter, which is a transcription initiation factor to which RNA polymerase binds, an arbitrary operator sequence for regulating transcription, a sequence encoding a suitable mRNA ribosome binding site, and transcription and translation. It may include a sequence controlling termination, a terminator, etc., more preferably, a 5' UTR region gene of M17 to increase the amount of protein synthesis, and minimize protein degradation so that the protein can be stably maintained in the endoplasmic reticulum. It may additionally include an HDEL gene, etc.
- a tag gene for recombinant protein production and more preferably, a tag gene for increasing the production of recombinant protein, a tag gene for maintaining structural stability of the recombinant protein, and a recombinant protein for easily separating the recombinant protein. It may additionally include a tag gene, a marker gene for selection such as an antibiotic resistance gene for selecting transformants, and the like.
- Representative tags for easy separation include Avi tag, Calmodulin tag, polyglutamate tag, and E tag.
- the selection marker genes typically include herbicide resistance genes such as glyphosate or phosphinothricin, kanamycin, G418, bleomycin, hygromycin ( hygromycin), antibiotic resistance genes such as chloramphenicol, and aadA genes.
- promoter examples include pEMU promoter, MAS promoter, histone promoter, Clp promoter, and cauliflower mosaic virus Origin 35S Promo cauliflower mosaic virus (cauliflower mosaic virus) derived 19S RNA promoter, plant actin protein promoter, ubiquitin protein promoter, CMV (Cytomegalovirus) promoter, SV40 (Simian virus 40) promoter, RSV (Respiratory syncytial virus) promoter, EF- 1 ⁇ (Elongation factor-1 alpha) promoter, etc.
- the terminator is typically nopaline synthase (NOS), rice amylase RAmy1 A terminator, phaseolin terminator, octopine of Agrobacterium tumefaciens (Octopine) gene terminator, E. coli's rrnB1/B2 terminator, etc., but the type of the added gene is not limited as long as it is a type conventionally used in the production of recombinant proteins.
- NOS nopaline synthase
- rice amylase RAmy1 A terminator phaseolin terminator
- Octopine octopine of Agrobacterium tumefaciens
- transgenic organism is a molecular genetic method that converts an external gene into an organism. It is a living organism prepared by injecting, preferably a living organism transformed by the recombinant expression vector of the present invention, and the living organism is not limited as long as it is a living organism such as a microorganism, eukaryotic cell, insect, animal, plant, etc., preferably Escherichia coli, salmonella, bacillus, yeast, animal cells, mice, rats, dogs, monkeys, pigs, horses, cows, Agrobacterium tumefaciens, plants, etc., but not limited thereto.
- the transformants are transformed ), transfection, Agrobacterium-mediated transformation method, particle gun bombardment, sonication, electroporation, PEG (Polyethylen glycol)-mediated It may be prepared by a method such as a transformation method, but there is no limitation as long as the vector can be injected.
- the FGF21 mutant protein may be thermostable or thermoelastic.
- Heat-Stability refers to the fact that protein denaturation or precipitation does not occur even when a certain amount of heat is applied under heat treatment conditions, and the physiological activity of proteins does not decrease or change. It refers to a property in which original characteristics/functions are maintained the same/similar to those before heat treatment.
- heat-elasticity used throughout the present specification means that the original properties/functions of proteins are restored to the same/similar to those before heat treatment when a certain level of heat is applied under heat treatment conditions and then re-cooled. it means.
- the FGF21 mutant protein may have a beta-trifoil ( ⁇ trefoil) structure.
- beta-trifoil ( ⁇ or "beta-trifoil fold” structure) refers to a protein folding structure consisting of six beta hairpins each formed from two beta strands. The structure is three hairpins , and the structure has approximately three-fold symmetry.
- the method may include heating the transformant or culture medium to a temperature higher than a temperature at which general proteins can be denatured, specifically, the temperature is 40 ° C. or higher, 45 ° C. or higher, It may be 50 ° C or more, 55 ° C or more, 60 ° C or more, 65 ° C or more, 70 ° C or more, 75 ° C or more, 80 ° C or more, 85 ° C or more, 90 ° C or more, 95 ° C or more, or 100 ° C, more specifically , 40 ° C or more to 300 ° C or less, 40 ° C or more to 250 ° C or less, 40 ° C or more to 200 ° C or less, 40 ° C or more to 150 ° C or less, 40 ° C or more to 120 ° C or less, 50 ° C or more to 300 ° C or less, 50 °C or more to 250 °C or less, 50 °C or more to 200 °C or less, 50 °C or or
- the FGF21 mutant protein may be additionally bound to a thermostable protein tag.
- thermostable protein tag may be linked to the N-terminus or C-terminus of the FGF21 variant protein, without affecting the characteristics/functions of the FGF21 variant protein, while producing/producing the protein. It is not limited thereto as long as the purification efficiency can be improved.
- the heat-stable protein tag is a heat-resistant protein tag that is not denatured even when heat is applied, and is a thermophilic organism-derived protein, thioredox
- thermostable proteins such as Thioredoxin (Trx) and DnaK.
- the FGF21 mutant protein and the protein tag may be directly linked or linked through a linker.
- the linker is not particularly limited as long as it does not affect the characteristics/functions of the FGF21 variant protein or the fusion protein to which the protein tag is linked, but may be composed of, for example, a nucleic acid or a peptide.
- the linker may be cleaved or degraded by various biological or chemical actions such as enzymatic action in an intracellular or external process. Specifically, it may be cleaved by protease treatment using tobacco etch virus (TEV) protease or a specific sequence recognized by proteases such as Thrombin and Factor Xa as a linker, or asparagine -It may be cleaved through heat treatment (40-42 ° C, 3 hours) using hydroxylamine (NH 2 OH) using a linker composed of glycine (Asn-Gly).
- TSV tobacco etch virus
- Thrombin and Factor Xa Thrombin and Factor Xa
- asparagine -It may be cleaved through heat treatment (40-42 ° C, 3 hours) using hydroxylamine (NH 2 OH) using a linker composed of glycine (Asn-Gly).
- the step of culturing the transformant is not particularly limited thereto, but may be performed by a known batch culture method, continuous culture method, fed-batch culture method, or the like.
- the culture conditions are not particularly limited thereto, but a basic compound (eg, sodium hydroxide, potassium hydroxide, or ammonia) or an acidic compound (eg, phosphoric acid or sulfuric acid) using a suitable pH (eg, pH 5 to 9, specifically can be adjusted to pH 6-8, most specifically pH 6.8), and oxygen or an oxygen-containing gas mixture can be introduced into the culture to maintain aerobic conditions.
- the culture temperature may be maintained at 20 to 45° C., specifically 25 to 40° C., and may be cultured for about 10 to 160 hours.
- the FGF21 mutant protein produced by the culture may be secreted into the medium or remain in the transformant.
- the culture medium used is sugar and carbohydrates (eg,
- fats and oils eg soybean oil, sunflower oil, peanut oil and coconut oil
- fatty acids eg palmitic acid, stearic acid and linoleic acid
- alcohols eg, glycerol and ethanol
- organic acids eg, acetic acid
- Nitrogen sources include nitrogen-containing organic compounds such as peptone, yeast extract, broth, malt extract, corn steep liquor, soybean meal and urea, or inorganic compounds such as ammonium sulfate, ammonium chloride, ammonium phosphate, ammonium carbonate and Ammonium nitrate) and the like may be used individually or in combination, but are not limited thereto.
- Potassium dihydrogen phosphate, dipotassium hydrogen phosphate, and sodium-containing salts corresponding thereto may be used individually or in combination as a phosphorus source, but are not limited thereto, and other metal salts (eg, magnesium sulfate or iron sulfate), amino acids, and vitamins Essential growth-promoting substances such as may be included.
- the method may further include removing proteins denatured by heat treatment.
- the step of removing the denatured protein may be to precipitate the protein denatured by heat treatment, specifically, the protein derived from the transformant through centrifugation, and then remove the precipitate.
- the FGF21 mutant protein which is not denatured by heat treatment, exists in the supernatant portion after centrifugation, and can be easily obtained.
- the method may further include recovering the produced FGF21 mutant protein.
- This recovery step may be a step of recovering from cultured cells or their supernatant, and a person skilled in the art can select an appropriate recovery process.
- the method for recovering the FGF21 protein produced in the culturing step can be used to collect the desired product from the culture medium using a suitable method known in the art according to a culture method, for example, a batch, continuous or fed-batch culture method. .
- proteins (proteins derived from the transformant) other than the FGF21 mutant protein precipitated by denaturation by heat treatment are removed using centrifugation, etc., and heat stability is maintained even after heat treatment. It may be to obtain only the FGF21 mutant protein that is not denatured due to (thermoelasticity).
- a method for purifying a FGF21 mutant protein comprising heating a mixture containing the FGF21 mutant protein and impurities to a temperature of 40° C. or higher.
- the impurities include proteins other than the FGF21 mutant protein generated in the FGF21 mutant protein production process, and may specifically include proteins derived from transformants that produce the FGF21 mutant protein. have.
- the step of purifying the FGF21 mutant protein is denatured by heat treatment, and precipitated impurities (proteins derived from the transformant) are removed using centrifugation, etc., and heat stability (thermal elasticity) is maintained even after heat treatment. It may be to obtain only the FGF21 mutant protein that is not denatured due to the
- Another aspect provides novel FGF21 variants.
- Another aspect provides a composition for treating metabolic or cardiovascular disorders comprising the novel FGF21 variant.
- the metabolic disorder or cardiovascular disorder is selected from hypercholesterolemia, dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), type 2 diabetes, and obesity. it may be
- the treatment, prevention, or management of the metabolic disorder or cardiovascular disorder reduces one or more of the subject's body weight, liver fat content, elevated LDL-C, total-C, triglycerides, and Apo B levels.
- the subject's body weight may include
- the metabolic disorder or cardiovascular disorder may be dyslipidemia, optionally mixed dyslipidemia, hypertriglyceridemia, optionally severe hypertriglyceridemia, or hypercholesterolemia, optionally primary hypercholesterolemia.
- the composition may be one or more of a pharmaceutical composition, a health functional food composition, and a feed composition.
- the pharmaceutical composition may further include suitable carriers, excipients or diluents commonly used in the preparation of pharmaceutical compositions.
- a composition containing a pharmaceutically acceptable carrier may be in various oral or parenteral formulations. When formulated, it may be prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.
- Solid preparations for oral administration may include tablet pills, powders, granules, capsules, etc., and such solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose in one or more compounds. It can be prepared by mixing lactose, gelatin, etc.
- Liquid preparations for oral administration include suspensions, internal solutions, emulsions, syrups, etc.
- various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included.
- Formulations for parenteral administration may include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations, and suppositories.
- Vegetable oils such as propylene glycol, polyethylene glycol, and olive oil may be used as non-aqueous solvents and suspending solvents.
- witepsol macrogol, tween 61, cacao butter, laurin paper, glycerogelatin, and the like may be used.
- the pharmaceutical composition is selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, internal solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. It can have any one formulation that is.
- Another aspect provides a method for treating metabolic disorders or cardiovascular disorders, comprising administering the pharmaceutical composition to a subject.
- the pharmaceutical composition is administered in a pharmaceutically effective amount.
- the term "administration” refers to introducing the pharmaceutical composition of the present invention to a subject by any suitable method, and the route of administration is through various oral or parenteral routes as long as it can reach the target tissue. It can be.
- composition of the present invention can be appropriately administered to a subject according to a conventional method, administration route, and dosage used in the art according to purpose or necessity.
- administration routes include oral, parenteral, subcutaneous, intraperitoneal, intrapulmonary, and intranasal administration
- parenteral injection includes intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.
- an appropriate dose and frequency of administration may be selected according to a method known in the art, and the amount and frequency of administration of the pharmaceutical composition of the present invention actually administered depend on the type of symptom to be treated, route of administration, gender, health It can be appropriately determined by various factors such as condition, diet, age and weight of the individual, and severity of the disease.
- pharmaceutically effective amount in the present invention means an amount sufficient to inhibit or alleviate metabolic disorders or cardiovascular disorders at a reasonable benefit / risk ratio applicable to medical use, and the effective dose level depends on the type and severity of the subject, Age, gender, activity of the drug, sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field.
- the term "subject” refers to all animals, including humans, that have or may develop metabolic disorders or cardiovascular disorders, and effectively prevent metabolic disorders or cardiovascular disorders by administering the pharmaceutical composition of the present invention to an object. can be cured
- Another aspect provides the use of the pharmaceutical composition for the treatment of metabolic disorders or cardiovascular disorders.
- the health functional food composition may be added to food as it is or used together with other foods or food ingredients, and may be appropriately used according to conventional methods.
- the health functional food composition is not particularly limited in other ingredients other than containing the active ingredient as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as additional ingredients like conventional beverages.
- Examples of the aforementioned natural carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrins, cyclodextrins, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol.
- natural flavoring agents thaumatin, stevia extract (eg rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can advantageously be used.
- the ratio of the natural carbohydrates can be appropriately determined by a person skilled in the art.
- the health functional food composition includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its It may contain salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, and the like. These components may be used independently or in combination. The ratio of these additives can also be appropriately selected by those skilled in the art.
- FGF21 is attracting attention from various pharmaceutical companies around the world as a candidate for the treatment of obesity, diabetes and nonalcoholic steatohepatitis (NASH).
- FGF21 is a protein that regulates glucose and lipid metabolism by influencing cell activity in brain, liver, pancreas, muscle, and adipose tissue.
- tyrosine kinase domain which is a cytosolic domain, signaling of intracellular kinases can be triggered.
- the FGF21 protein consists of 209 amino acids, and a new variant was designed based on a form in which four amino acids at the N-terminus were removed from the mature form (29-209aa) in which the signal peptide (1-28aa) was removed, In order to develop novel FGF21 variants, FGF21 variants were designed with the following strategy.
- the engineering position was selected through analysis of the crystal structure of the complex of hFGF21CT and ⁇ . Based on the structural analysis, it was possible to identify three regions (Ser195 & Pro199 & Ser200) in which the residues in the sequence of FGF21CT do not participate in the interaction with ⁇ -Klotho. Therefore, for the purpose of enhancing the activity and improving the stability of FGF21, mutation 2 (Ser195Gly & Pro199Gly & Ser200Gly) in which glycine was introduced was developed. This is a protein structure-based engineering strategy designed to help the interaction with ⁇ -Klotho and at the same time increase resistance from protease attack.
- Trx-FGF21 The following heat-resistant protein was used as a tag, and E. coli overexpressing Trx-FGF21 in which Trx was fused to a FGF21 mutant was disrupted by sonication and then centrifuged. The supernatant solution containing Trx-FGF21 was incubated at high temperature (50-100° C.) for 10 minutes, and then proteins denatured at high temperature and Trx-FGF21 were separated by centrifugation (Fig. 4, left).
- cleavable linker In addition, in order to obtain a pure FGF21 mutant from which Trx has been removed, a cleavable linker must be inserted between the tag and the FGF21 mutant.
- Tobaccoetch virus (TEV)-derived proteases or specific sequences recognized by proteases such as Thrombin and Factor Xa can be used as linkers.
- FGF21 can be separated from the tag by protease, and can then be purified with a high purity of 90% or more through two or less chromatographic techniques (Fig. 4, right).
- FGF21 (FGF21 ⁇ N4, 33-209aa) from which the signal peptide (1-28aa) and N-terminal 4 amino acids (29-32aa) of FGF21 are additionally removed is produced through heat treatment, and commercially sold FGF21 (29-209aa) and activity were compared.
- FGF21 binds to FGF receptors together with beta-klotho, a coreceptor, and activates FGFR1c, one of the FGF receptors, to exhibit various physiological effects. Therefore, HEK293 cells expressing FGFR1c and beta-Klotho and equipped with a reporter system capable of confirming the degree of FGFR1c activation were used to confirm the FGF receptor activation ability by FGF21.
- FGF21 has no effect on the function of activating FGFR1c.
- the cell activity improvement effect of the highly purified FGF21 variants 1, 2, and 3 in Example 2 was measured through an in vitro cell assay.
- FGFR signaling in adipocytes by FGF21 wild-type and variants was confirmed by western blot.
- ERK is one of the representative signal transducers of the intracellular signal transduction system following FGFR activation, and becomes phosphorylated (pERK) according to FGFR activation.
- pERK phosphorylated
- variant 3 a novel FGF21 variant developed in the present invention, has excellent activity and can be selected as an FGF21 biobetter.
Abstract
Description
Claims (17)
- FGF21 변이체를 포함하는 폴리펩타이드로서, 상기 변이체는 야생형 FGF21 단백질에 하나 이상의 돌연변이가 포함된 것을 특징으로 하는 폴리펩타이드.
- 청구항 1에 있어서,상기 돌연변이는 아미노산의 결실, 치환 및 부가 중 하나 이상인 것인, 폴리펩타이드.
- 청구항 1에 있어서,상기 야생형 FGF21 단백질은 신호 펩타이드가 제거된 성숙된 형태인 것인, 폴리펩타이드.
- 청구항 1에 있어서,상기 야생형 FGF21 단백질은 서열번호 2의 아미노산 서열을 포함하는 것인, 폴리펩타이드.
- 청구항 1에 있어서,상기 FGF21 변이체는 N-말단의 1 내지 4개의 아미노산이 결실된 것인, 폴리펩타이드.
- 청구항 1에 있어서,상기 FGF21 변이체는 Q184E, S195G, P199G, S200G, Q201H 및 A208T 로 구성된 군에서 선택된 하나 이상으로 치환된 것인, 폴리펩타이드.
- 청구항 1에 있어서,상기 FGF21 변이체는 S195G, P199G, S200G, Q201H 및 A208T로 치환된 것인, 폴리펩타이드.
- 청구항 1에 있어서,상기 FGF21 변이체는 서열번호 3 내지 6의 아미노산 서열 중 하나인 것인, 폴리펩타이드.
- 청구항 1에 있어서,상기 FGF21 변이체는 야생형에 비해 활성이 향상된 것인, 폴리펩타이드.
- FGF21 변이체 단백질을 생산하는 형질전환체를 배양하는 단계; 및상기 형질전환체 또는 배양액을 40℃ 이상의 온도로 가열하는 단계를 포함하는, FGF21 변이체 단백질의 생산 방법.
- 청구항 10에 있어서,상기 FGF21 변이체 단백질은 열안정성 또는 열탄력성이 있는 것인, FGF21 변이체 단백질의 생산 방법.
- 청구항 10에 있어서,상기 FGF21 변이체 단백질은 베타-트리포일(β-trefoil) 구조를 포함하고 있는 것인, FGF21 변이체 단백질의 생산 방법.
- 청구항 10에 있어서,상기 FGF21 변이체 단백질은 열안정성 단백질 태그(tag)가 추가로 결합되어 있는 것인, FGF21 변이체 단백질의 생산 방법.
- 청구항 10에 있어서,상기 방법은 열처리에 의해 변성된 단백질을 제거하는 단계를 추가로 포함하는 것인, FGF21 변이체 단백질의 생산 방법.
- FGF21 변이체 단백질 및 불순물이 포함된 혼합물을 40℃ 이상의 온도로 가열하는 단계를 포함하는, FGF21 변이체 단백질의 정제 방법.
- 청구항 15에 있어서,상기 방법은 열처리에 의해 변성된 단백질을 제거하는 단계를 추가로 포함하는 것인, FGF21 변이체 단백질의 정제 방법.
- FGF21 변이체를 포함하는, 대사 장애 또는 심혈관 장애 치료용 약학적 조성물.
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP22820595.1A EP4353742A1 (en) | 2021-06-10 | 2022-06-10 | Development of novel fgf21 variant, and production technique and use thereof |
CN202280041814.2A CN117480177A (zh) | 2021-06-10 | 2022-06-10 | 新型fgf21变体的开发及其生产技术和用途 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020210075460A KR102631925B1 (ko) | 2021-06-10 | 2021-06-10 | 신규 fgf21 변이체 개발 및 이의 생산기법과 용도 |
KR10-2021-0075460 | 2021-06-10 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2022260468A1 true WO2022260468A1 (ko) | 2022-12-15 |
Family
ID=84426238
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/KR2022/008202 WO2022260468A1 (ko) | 2021-06-10 | 2022-06-10 | 신규 fgf21 변이체 개발 및 이의 생산기법과 용도 |
Country Status (4)
Country | Link |
---|---|
EP (1) | EP4353742A1 (ko) |
KR (1) | KR102631925B1 (ko) |
CN (1) | CN117480177A (ko) |
WO (1) | WO2022260468A1 (ko) |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20150006060A (ko) * | 2012-06-11 | 2015-01-15 | 일라이 릴리 앤드 캄파니 | 섬유모세포 성장 인자 21 단백질 |
KR20170095256A (ko) * | 2014-12-23 | 2017-08-22 | 노보 노르디스크 에이/에스 | Fgf21 유도체 및 그것의 용도 |
KR20180083938A (ko) * | 2015-12-02 | 2018-07-23 | 사노피 | Fgf21 변이체 |
KR102217254B1 (ko) * | 2011-09-26 | 2021-02-19 | 노파르티스 아게 | 대사 장애를 치료하기 위한 이중 기능 단백질 |
KR20210075460A (ko) | 2019-12-13 | 2021-06-23 | 주식회사 피티지 | 고토크 다상 ipm bldc 모터 제어장치 |
KR20220059660A (ko) * | 2020-11-03 | 2022-05-10 | 주식회사 도어코코리아 | Fgf21의 열탄력성을 이용한 fgf21 생산 방법 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2019123427A1 (en) * | 2017-12-22 | 2019-06-27 | Novartis Ag | Methods of treating metabolic disorders with fgf21 variants |
-
2021
- 2021-06-10 KR KR1020210075460A patent/KR102631925B1/ko active IP Right Grant
-
2022
- 2022-06-10 EP EP22820595.1A patent/EP4353742A1/en active Pending
- 2022-06-10 WO PCT/KR2022/008202 patent/WO2022260468A1/ko active Application Filing
- 2022-06-10 CN CN202280041814.2A patent/CN117480177A/zh active Pending
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR102217254B1 (ko) * | 2011-09-26 | 2021-02-19 | 노파르티스 아게 | 대사 장애를 치료하기 위한 이중 기능 단백질 |
KR20150006060A (ko) * | 2012-06-11 | 2015-01-15 | 일라이 릴리 앤드 캄파니 | 섬유모세포 성장 인자 21 단백질 |
KR20170095256A (ko) * | 2014-12-23 | 2017-08-22 | 노보 노르디스크 에이/에스 | Fgf21 유도체 및 그것의 용도 |
KR20180083938A (ko) * | 2015-12-02 | 2018-07-23 | 사노피 | Fgf21 변이체 |
KR20210075460A (ko) | 2019-12-13 | 2021-06-23 | 주식회사 피티지 | 고토크 다상 ipm bldc 모터 제어장치 |
KR20220059660A (ko) * | 2020-11-03 | 2022-05-10 | 주식회사 도어코코리아 | Fgf21의 열탄력성을 이용한 fgf21 생산 방법 |
Non-Patent Citations (2)
Title |
---|
"Current protocols in protein science", WILEY , US , article JOHN MCCOY, EDWARD LAVALLIE: "Expression and Purification of Thioredoxin Fusion Proteins", pages: 16.8.1 - 16.8.14, XP009151941, DOI: 10.1002/0471142727.mb1608s28 * |
JUNG YE-EUN, LEE KYEONG WON, CHO JAE HYUN, BAE DA-WOON, JEONG BO-GYEONG, JUNG YEON-JU, AN YOUNG JUN, KIM KYUNGCHAN, LEE GA SEUL, K: "Temperature-responsive structural reversibility of FGF21 and structure-based design of its variant with enhanced potency", BIORXIV, 29 November 2021 (2021-11-29), pages 1 - 32, XP093014596, DOI: 10.1101/2021.11.16.468794 * |
Also Published As
Publication number | Publication date |
---|---|
EP4353742A1 (en) | 2024-04-17 |
KR102631925B1 (ko) | 2024-02-01 |
KR20220166554A (ko) | 2022-12-19 |
CN117480177A (zh) | 2024-01-30 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
WO2018030608A1 (ko) | Cas9 단백질 및 가이드 RNA의 혼성체를 함유하는 나노 리포좀 전달체 조성물 | |
US4431740A (en) | DNA Transfer vector and transformed microorganism containing human proinsulin and pre-proinsulin genes | |
CS219257B2 (en) | Method of making the eukaryotic protein | |
WO2023124847A1 (zh) | 一种长效glp-1衍生物 | |
EA008831B1 (ru) | Слитые белки аналогов glp-1 | |
KR20070091131A (ko) | 카복시-말단 아미드화 펩티드를 제조하는 방법 | |
WO2014208970A1 (ko) | 트랜스케톨라아제 유전자 프로모터 변이체 및 이의 용도 | |
WO2012030068A2 (ko) | 1-데옥시노지리마이신 합성관련 폴리펩타이드 및 이의 용도 | |
JP7097434B2 (ja) | ヒトfgf21変異体、その製造方法、及びその使用 | |
WO2015009074A2 (ko) | 신규 변이 오르니틴 디카복실레이즈 단백질 및 이의 용도 | |
JP2637393B2 (ja) | プロインシュリンおよびプレプロインシュリン産生暗号を有するヒト遺伝子 | |
CN113735960B (zh) | 一种fgf重组蛋白治疗nash的应用 | |
WO2022260468A1 (ko) | 신규 fgf21 변이체 개발 및 이의 생산기법과 용도 | |
WO2022097914A1 (ko) | 에프지에프21의 열탄력성을 이용한 에프지에프21 생산 방법 | |
JPH03503596A (ja) | 蛋白質若しくはポリペプチドの調製方法、該ポリペプチドをコードするdna配列、該dna配列およびポリペプチドを有する微生物および該ポリペプチドの薬学的製剤としての利用 | |
WO2013129852A1 (ko) | 신규한 인터루킨-6에 대한 리피바디 및 그 용도 | |
JP2862870B2 (ja) | 新規ペプチド | |
WO2020101366A1 (en) | Activin/bmp7 chimeras: super-active sab704 and sab715, and their respective noggin-sensitized variants, nab704 and nab715; and nab204 | |
WO2019098708A1 (ko) | 해조류 유래 아가로트리오스의 제조방법 및 프리바이오틱으로서의 용도 | |
WO2018004294A9 (ko) | 인간 성장호르몬 변이 단백질 또는 이의 트랜스페린 융합 단백질을 유효성분으로 포함하는 약학적 조성물 | |
WO2022010319A1 (ko) | 글루카곤-유사 펩타이드-1 및 인터루킨-1 수용체 길항제를 포함하는 융합단백질 및 이의 용도 | |
WO2021136521A1 (zh) | 多肽及其应用 | |
WO2017065529A1 (ko) | O-아세틸호모세린 설피드릴라제 변이체 및 이를 이용한 l-메치오닌 제조 방법 | |
CN113018424B (zh) | Cst1在预防和/或治疗肝脏免疫失调疾病中的应用 | |
WO2016018133A9 (ko) | 보체 단백질 C5a와 결합할 수 있는 신규한 폴리펩타이드 및 그 용도 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 22820595 Country of ref document: EP Kind code of ref document: A1 |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2022820595 Country of ref document: EP |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
ENP | Entry into the national phase |
Ref document number: 2022820595 Country of ref document: EP Effective date: 20240110 |
|
ENP | Entry into the national phase |
Ref document number: 2022820595 Country of ref document: EP Effective date: 20240110 |