CN114249837B - 一种多肽、及其制备方法和应用 - Google Patents
一种多肽、及其制备方法和应用 Download PDFInfo
- Publication number
- CN114249837B CN114249837B CN202111623326.6A CN202111623326A CN114249837B CN 114249837 B CN114249837 B CN 114249837B CN 202111623326 A CN202111623326 A CN 202111623326A CN 114249837 B CN114249837 B CN 114249837B
- Authority
- CN
- China
- Prior art keywords
- polypeptide
- at1r
- tat
- seq
- protein
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 61
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 58
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 58
- 238000002360 preparation method Methods 0.000 title abstract description 7
- 208000031225 myocardial ischemia Diseases 0.000 claims abstract description 20
- 208000007201 Myocardial reperfusion injury Diseases 0.000 claims abstract description 18
- 101150059573 AGTR1 gene Proteins 0.000 claims abstract description 15
- 125000000539 amino acid group Chemical group 0.000 claims abstract description 9
- 230000002107 myocardial effect Effects 0.000 claims abstract description 9
- 208000028867 ischemia Diseases 0.000 claims abstract description 8
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims abstract description 6
- 206010063837 Reperfusion injury Diseases 0.000 claims abstract description 5
- 108090000623 proteins and genes Proteins 0.000 claims description 13
- 102000004169 proteins and genes Human genes 0.000 claims description 13
- 239000003814 drug Substances 0.000 claims description 12
- 150000007523 nucleic acids Chemical class 0.000 claims description 12
- 230000002633 protecting effect Effects 0.000 claims description 12
- 239000001963 growth medium Substances 0.000 claims description 10
- 230000014509 gene expression Effects 0.000 claims description 9
- 108020004707 nucleic acids Proteins 0.000 claims description 9
- 102000039446 nucleic acids Human genes 0.000 claims description 9
- 108020004414 DNA Proteins 0.000 claims description 7
- 102000037865 fusion proteins Human genes 0.000 claims description 7
- 108020001507 fusion proteins Proteins 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 7
- 239000002299 complementary DNA Substances 0.000 claims description 6
- 230000000694 effects Effects 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 5
- 238000001976 enzyme digestion Methods 0.000 claims description 5
- 239000013604 expression vector Substances 0.000 claims description 5
- 239000002773 nucleotide Substances 0.000 claims description 5
- 125000003729 nucleotide group Chemical group 0.000 claims description 5
- 102000053602 DNA Human genes 0.000 claims description 4
- 102000004190 Enzymes Human genes 0.000 claims description 4
- 108090000790 Enzymes Proteins 0.000 claims description 4
- 108091026890 Coding region Proteins 0.000 claims description 3
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 3
- 239000007924 injection Substances 0.000 claims description 3
- 238000002347 injection Methods 0.000 claims description 3
- 239000012139 lysis buffer Substances 0.000 claims description 3
- 238000001742 protein purification Methods 0.000 claims description 3
- 238000010839 reverse transcription Methods 0.000 claims description 3
- 238000012216 screening Methods 0.000 claims description 3
- 241000672609 Escherichia coli BL21 Species 0.000 claims description 2
- 108010076818 TEV protease Proteins 0.000 claims description 2
- 239000004480 active ingredient Substances 0.000 claims description 2
- 239000007925 intracardiac injection Substances 0.000 claims description 2
- 239000007927 intramuscular injection Substances 0.000 claims description 2
- 238000010255 intramuscular injection Methods 0.000 claims description 2
- 239000007928 intraperitoneal injection Substances 0.000 claims description 2
- 238000010253 intravenous injection Methods 0.000 claims description 2
- 239000007929 subcutaneous injection Substances 0.000 claims description 2
- 238000010254 subcutaneous injection Methods 0.000 claims description 2
- 239000000825 pharmaceutical preparation Substances 0.000 claims 3
- 229940127557 pharmaceutical product Drugs 0.000 claims 3
- 238000004519 manufacturing process Methods 0.000 claims 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 238000010276 construction Methods 0.000 claims 1
- 230000001939 inductive effect Effects 0.000 claims 1
- 229940054733 arestin Drugs 0.000 abstract description 25
- 150000001413 amino acids Chemical group 0.000 abstract description 14
- 230000003993 interaction Effects 0.000 abstract description 5
- CUKWUWBLQQDQAC-VEQWQPCFSA-N (3s)-3-amino-4-[[(2s)-1-[[(2s)-1-[[(2s)-1-[[(2s,3s)-1-[[(2s)-1-[(2s)-2-[[(1s)-1-carboxyethyl]carbamoyl]pyrrolidin-1-yl]-3-(1h-imidazol-5-yl)-1-oxopropan-2-yl]amino]-3-methyl-1-oxopentan-2-yl]amino]-3-(4-hydroxyphenyl)-1-oxopropan-2-yl]amino]-3-methyl-1-ox Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C)C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@@H](N)CC(O)=O)C(C)C)C1=CC=C(O)C=C1 CUKWUWBLQQDQAC-VEQWQPCFSA-N 0.000 abstract description 2
- 102000005862 Angiotensin II Human genes 0.000 abstract description 2
- 101800000733 Angiotensin-2 Proteins 0.000 abstract description 2
- 108010062481 Type 1 Angiotensin Receptor Proteins 0.000 abstract description 2
- 229950006323 angiotensin ii Drugs 0.000 abstract description 2
- 102000005962 receptors Human genes 0.000 abstract description 2
- 108020003175 receptors Proteins 0.000 abstract description 2
- 102000010913 Type 1 Angiotensin Receptor Human genes 0.000 abstract 1
- 230000002265 prevention Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 18
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 9
- 230000006870 function Effects 0.000 description 7
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 6
- 230000010410 reperfusion Effects 0.000 description 6
- 210000004413 cardiac myocyte Anatomy 0.000 description 5
- 230000000149 penetrating effect Effects 0.000 description 5
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 4
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 208000037891 myocardial injury Diseases 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 3
- 108090000045 G-Protein-Coupled Receptors Proteins 0.000 description 3
- 102000003688 G-Protein-Coupled Receptors Human genes 0.000 description 3
- 206010021143 Hypoxia Diseases 0.000 description 3
- 210000000170 cell membrane Anatomy 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 230000007954 hypoxia Effects 0.000 description 3
- 230000019491 signal transduction Effects 0.000 description 3
- 238000011144 upstream manufacturing Methods 0.000 description 3
- 101150028074 2 gene Proteins 0.000 description 2
- 241000723873 Tobacco mosaic virus Species 0.000 description 2
- 230000030833 cell death Effects 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 210000005003 heart tissue Anatomy 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229910052759 nickel Inorganic materials 0.000 description 2
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- DYJJJCHDHLEFDW-FXQIFTODSA-N Ala-Pro-Cys Chemical compound C[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CS)C(=O)O)N DYJJJCHDHLEFDW-FXQIFTODSA-N 0.000 description 1
- IASNWHAGGYTEKX-IUCAKERBSA-N Arg-Arg-Gly Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)NCC(O)=O IASNWHAGGYTEKX-IUCAKERBSA-N 0.000 description 1
- ATABBWFGOHKROJ-GUBZILKMSA-N Arg-Pro-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O ATABBWFGOHKROJ-GUBZILKMSA-N 0.000 description 1
- LFWOQHSQNCKXRU-UFYCRDLUSA-N Arg-Tyr-Phe Chemical compound C([C@H](NC(=O)[C@H](CCCN=C(N)N)N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=C(O)C=C1 LFWOQHSQNCKXRU-UFYCRDLUSA-N 0.000 description 1
- PQKSVQSMTHPRIB-ZKWXMUAHSA-N Asn-Val-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O PQKSVQSMTHPRIB-ZKWXMUAHSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 238000011746 C57BL/6J (JAX™ mouse strain) Methods 0.000 description 1
- 206010007559 Cardiac failure congestive Diseases 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 206010019280 Heart failures Diseases 0.000 description 1
- 208000013875 Heart injury Diseases 0.000 description 1
- CWSZWFILCNSNEX-CIUDSAMLSA-N His-Ser-Asn Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)N)C(=O)O)N CWSZWFILCNSNEX-CIUDSAMLSA-N 0.000 description 1
- 108700000788 Human immunodeficiency virus 1 tat peptide (47-57) Proteins 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- FQYQMFCIJNWDQZ-CYDGBPFRSA-N Ile-Pro-Pro Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 FQYQMFCIJNWDQZ-CYDGBPFRSA-N 0.000 description 1
- LOLUPZNNADDTAA-AVGNSLFASA-N Leu-Gln-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O LOLUPZNNADDTAA-AVGNSLFASA-N 0.000 description 1
- ONPJGOIVICHWBW-BZSNNMDCSA-N Leu-Lys-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 ONPJGOIVICHWBW-BZSNNMDCSA-N 0.000 description 1
- PPGBXYKMUMHFBF-KATARQTJSA-N Leu-Ser-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PPGBXYKMUMHFBF-KATARQTJSA-N 0.000 description 1
- SQUFDMCWMFOEBA-KKUMJFAQSA-N Leu-Ser-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 SQUFDMCWMFOEBA-KKUMJFAQSA-N 0.000 description 1
- WSXTWLJHTLRFLW-SRVKXCTJSA-N Lys-Ala-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O WSXTWLJHTLRFLW-SRVKXCTJSA-N 0.000 description 1
- ATNKHRAIZCMCCN-BZSNNMDCSA-N Lys-Lys-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)N ATNKHRAIZCMCCN-BZSNNMDCSA-N 0.000 description 1
- YXPJCVNIDDKGOE-MELADBBJSA-N Lys-Lys-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)N)C(=O)O YXPJCVNIDDKGOE-MELADBBJSA-N 0.000 description 1
- MIFFFXHMAHFACR-KATARQTJSA-N Lys-Ser-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CCCCN MIFFFXHMAHFACR-KATARQTJSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000016943 Muramidase Human genes 0.000 description 1
- 108010014251 Muramidase Proteins 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 1
- XZFYRXDAULDNFX-UHFFFAOYSA-N N-L-cysteinyl-L-phenylalanine Natural products SCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XZFYRXDAULDNFX-UHFFFAOYSA-N 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- LWPMGKSZPKFKJD-DZKIICNBSA-N Phe-Glu-Val Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O LWPMGKSZPKFKJD-DZKIICNBSA-N 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- PYTKULIABVRXSC-BWBBJGPYSA-N Ser-Ser-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PYTKULIABVRXSC-BWBBJGPYSA-N 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- 102100026803 Type-1 angiotensin II receptor Human genes 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 230000009091 contractile dysfunction Effects 0.000 description 1
- 210000004351 coronary vessel Anatomy 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000012149 elution buffer Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 108010015792 glycyllysine Proteins 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 108010031424 isoleucyl-prolyl-proline Proteins 0.000 description 1
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 1
- 108010047926 leucyl-lysyl-tyrosine Proteins 0.000 description 1
- 229960000274 lysozyme Drugs 0.000 description 1
- 239000004325 lysozyme Substances 0.000 description 1
- 235000010335 lysozyme Nutrition 0.000 description 1
- 108010057952 lysyl-phenylalanyl-lysine Proteins 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 239000002808 molecular sieve Substances 0.000 description 1
- 230000008065 myocardial cell damage Effects 0.000 description 1
- 230000003680 myocardial damage Effects 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 230000007310 pathophysiology Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000004017 serum-free culture medium Substances 0.000 description 1
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 239000013598 vector Substances 0.000 description 1
- 239000011534 wash buffer 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
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/72—Receptors; Cell surface antigens; Cell surface determinants for hormones
- C07K14/723—G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH receptor
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- A61K48/005—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- 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
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/70—Vectors or expression systems specially adapted for E. coli
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/10—Fusion polypeptide containing a localisation/targetting motif containing a tag for extracellular membrane crossing, e.g. TAT or VP22
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Epidemiology (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Heart & Thoracic Surgery (AREA)
- Vascular Medicine (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Plant Pathology (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Urology & Nephrology (AREA)
- Cardiology (AREA)
- Endocrinology (AREA)
- Cell Biology (AREA)
- Immunology (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Dermatology (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明公开了一种多肽、及其制备方法和应用,该多肽名称为TAT‑AT1R‑1,为如下(1)或(2)所述的多肽:(1)SEQ ID NO:1所示的氨基酸序列第12‑65位氨基酸残基的多肽;(2)SEQ ID NO:1所示的氨基酸序列的多肽。本发明提供的多肽TAT‑AT1R‑1来源于血管紧张素II 1型受体(angiotensin II type1receptor,AT1R)羧基末端,TAT‑AT1R‑1能够代替AT1R与β‑arrestin2发生相互作用,进而保护心肌细胞抵御缺血/再灌注损伤。本发明的多肽TAT‑AT1R‑1在心肌缺血/再灌注损伤的治疗和预防中具有临床应用潜力。
Description
技术领域
本发明涉及医药技术领域,具体涉及一种多肽、及其制备方法和应用。
背景技术
血管紧张素II 1型受体(angiotensin II type 1receptor,AT1R)是一种典型的G蛋白偶联受体,其含有7次跨膜的空间结构,AT1R的持续激活会造成心脏的伤害,而其药理上的阻断则能够保护充血性心力衰竭和高血压等疾病。在心肌细胞损伤的病理过程中,AT1R和β-arrestin2的相互作用发挥了关键作用。
研究显示:心肌缺血/再灌注损伤状态,β-arrestin2表达量特异性增加,而β-arrestin1表达量则未发生明显改变,随后证实β-arrestin2是心肌缺血/再灌注导致心肌细胞死亡和心肌损伤的重要参与者,是关键的致病因素之一。过表达β-arrestin2能够诱发缺血/再灌注过程中心肌细胞死亡增加和可收缩性功能异常。过表达β-arrestin2的转基因动物缺血/再灌注造成的心肌损伤明显加重。而敲除β-arrestin2基因则能够有效保护心脏减轻缺血/再灌注的损伤。
AT1R作为典型的G蛋白偶联受体,其功能受到β-arrestin2的调控,两者这种典型相互作用是AT1R在心肌细胞病理生理过程中发挥作用的关键,如果以AT1R羧基末端氨基酸序列为基础设计多肽与β-arrestin2相互作用,或者能够抑制其后续信号传导,达到类似敲除β-arrestin2基因的效果——有效保护心肌细胞抵御缺血/再灌注损伤。
考虑到AT1R与β-arrestin2的相互作用是在细胞质内发生的,再考虑到GPCR羧基末端氨基酸序列在与β-arrestins相互作用过程中的关键功能,我们以AT1R羧基末端氨基酸序列为基础设计多肽,该多肽将竞争性地与β-arrestin2结合,阻断其介导的信号通路。同时,在多肽的氨基末端加入TAT穿膜肽,赋予其穿过细胞膜进入细胞的生物学功能。
目前干预心肌缺血/再灌注损伤的手段比较有限,大多是化学药物例如中国专利CN 104983731 B,但其选择性不高,毒副作用仍较大,生物药例如CN102558357B,仅为理论研究,但是却并没有临床应用价值,因此迫切需要新的保护心肌缺血/再灌注损伤的生物药物。
总之,研究和开发源于AT1R氨基酸序列的多肽药物,抑制β-arrestin2信号通路,将具有良好的保护心肌缺血/再灌注损伤的功效,具有临床应用前景。
发明内容
针对上述存在的技术不足,本发明的目的是提供一种多肽、及其制备方法和应用,该多肽具有保护心肌缺血/再灌注损伤作用的多肽,能够竞争性占据β-arrestin2功能区域,从而阻断β-arrestin2信号通路,达到保护心肌缺血/再灌注损伤的目的。
为实现上述目的,本发明提供一种具有保护心肌缺血/再灌注损伤作用的多肽,名称为TAT-AT1R-1,为如下(1)或(2)的多肽:
(1)SEQ ID NO:1所示的氨基酸序列第12-65位氨基酸残基的多肽;
(2)SEQ ID NO:1所示的氨基酸序列的多肽。
其中,序列表中SEQ ID NO:1所示的氨基酸序列由65个氨基酸残基组成,序列1中第1-11位氨基酸残基组成具有穿透细胞膜作用的区域,该区域和12-65位氨基酸前后位置可以互换,并且可以被具有相同作用的其他序列替换;序列1中第12-65位氨基酸残基构成能够和β-arrestin2蛋白质相互作用的区域。
本发明的第二个目的是提供所述一种多肽的制备方法为:
S1:构建表达载体,筛选重组子提取AC16心肌细胞总RNA,反转录得到cDNA,以cDNA为模板通过PCR获得AT1R羧基末端多肽的核酸序列,在引物中插入TAT编码序列和酶切位点,酶切连接法构建表达载体pWaldo-TAT-AT1R-1,转化大肠杆菌BL21表达菌株,筛选重组子;
S2:获得高纯度的TAT-AT1R-1和GFP融合蛋白接种重组子菌株于培养基中,诱导蛋白质表达,继续培养后离心收集细胞用于蛋白质纯化;将细胞悬浮于裂解缓冲液中,破碎细胞后去除细胞碎片;进一步纯化蛋白质可获得较高纯度的融合蛋白;
S3:纯化得到TAT-AT1R-1多肽将S2得到的蛋白质样品中加入TEV蛋白酶,酶切过夜,去掉没有酶切完全的融合蛋白和酶切后的GFP,最终收集得到纯化的TAT-AT1R-1多肽样品,浓缩备用。
本发明的第三个目的是提供编码所述多肽的核酸分子。
为实现上述目的,本发明所提供的编码所述多肽的核酸分子,为如下(3)或(4)的DNA分子:
(3)SEQ ID NO:2自5’末端起第34位至第198位核苷酸所示的DNA分子;
(4)SEQ ID NO:2所示的DNA分子。
其中,序列表中SEQ ID NO:2所示的核苷酸序列的长度为198bp,其编码氨基酸序列是序列表中SEQ ID NO:1所示的多肽片段。
本发明的第四个目的是提供含有所述多肽的编码核酸分子的表达盒、重组载体或重组菌。
本发明的第五个目的是提供所述多肽或所述核酸分子在制备保护心肌缺血/再灌注损伤作用的药品中的应用。
进一步地,所述保护心肌缺血/再灌注损伤作用包括减少心肌细胞死亡和和心肌损伤等。
本发明还提供一种具有保护心肌缺血/再灌注损伤作用的药品,其活性成分为上述的多肽或上述的核酸分子。
进一步地,所述药品的给药方式包括肌肉注射、静脉注射、心内注射、皮内注射、皮下注射、腹腔注射。
本发明的有益效果在于:本发明提供的多肽TAT-AT1R-1来源于AT1R羧基末端,所述多肽TAT-AT1R-1能够与β-arrestin2相互作用,竞争性占据β-arrestin2功能区域,从而阻断β-arrestin2信号通路,达到保护心肌缺血/再灌注损伤的目的。本发明的多肽TAT-AT1R-1在心肌缺血/再灌注损伤的预防和治疗中具有临床应用潜力。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本发明的一些实施例,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。
图1为肽段序列对缺氧/复氧造成心肌细胞AC16损伤的保护作用检测。
图2为肽段序列对心肌缺血/再灌注损伤的保护作用检测,其中A为对照组,B为多肽组。
具体实施方式
下面结合具体实例对本发明的发明方法进行详细描述和说明。其内容是对本发明的解释而非限定本发明的保护范围。
下面结合具体实施例对本发明进行进一步的详细描述,给出的实施例仅为了阐明本发明,而不是为了限制本发明的范围。本实施例中的定量实验,如无特殊说明均设置三次重复,结果取平均值。
下述实施例中的实验方法,如无特殊说明,均为常规的分子生物学方法。
下述实施例中所用的材料、试剂等,如无特殊说明,均可从商业途径得到。
下述实施例中人源心肌AC16细胞来源于ATCC(美国模式培养物集存库)。
实施例1多肽TAT-AT1R-1的获得
(1)具有穿模活性的TAT-AT1R-1多肽获得
提取AC16心肌细胞总RNA,反转录得到cDNA,以cDNA为模板通过PCR获得AT1R羧基末端多肽的核酸序列,其上游引物:CCATCTCGAGATGTACGGTCGTAAAAAACGTCGTCAGCGTCGTCGTGGGAAAAAATTTAAAAGATATTTTCTCC(SEQ ID NO.3),下游引物5'CGTCGGATCCCTCAACCTCAAAACATGGTGCAG 3'(SEQ ID NO.4),在上游引物中插入TAT编码序列,在上游和下游引物中分别加入XhoI和BamHI酶切位点,通过酶切连接法,构建表达载体pWaldo-TAT-AT1R-1,转化大肠杆菌BL21(DE3)表达菌株,筛选重组子。
接种重组子菌株于200ml LB培养基中,37℃培养过夜,转种100ml过夜培养物至2LLB培养基中,37℃培养至OD600 0.5~0.6之间,加入终浓度0.1mM的IPTG诱导蛋白质表达,20℃继续培养22h,6000g离心15min收集细胞用于蛋白质纯化。
将细胞悬浮于100ml裂解缓冲液中(50mM Tris-HCl,pH 7.5;300mM NaCl和5%甘油),加入50μg/mL溶菌酶,200U DnaseI和100mM PMSF,800bar破碎细胞,10,000rpm,4℃离心30min去除细胞碎片;将上清过镍柱,冲洗缓冲液(50mM Tris-HCL,pH 7.5,300mM NaCl,5%甘油,30mM咪唑)洗80ml去除杂蛋白,洗脱缓冲液(50mM Tris-HCL,pH7.5,300mM NaCl,5%甘油和300mM咪唑)洗脱,收集蛋白质,通过分子筛柱子(缓冲液:50mM Tris-HCL,pH7.5,300mM NaCl,5%甘油)进一步纯化,可获得较高纯度的TAT-AT1R-1和GFP融合蛋白。
为了进一步获得TAT-AT1R-1多肽,蛋白质样品中加入100μL浓度为3mg/mL的TEV(烟草花叶病毒)蛋白酶,4℃酶切过夜,样品重新通过镍柱,将没有酶切完全的融合蛋白和酶切后的GFP组分去掉。最终收集的TAT-AT1R-1多肽样品,通过截留分子量1,000的浓缩管,浓缩至5mg/mL左右,用于功能研究。纯化的多肽TAT-AT1R-1氨基酸序列如SEQ ID NO.1所示,其编码的核苷酸序列如SEQ ID NO.2所示。
其中,序列表中SEQ ID NO:1所示的氨基酸序列由65个氨基酸残基组成,序列1中第1-11位氨基酸残基组成具有穿透细胞膜作用的区域,该区域和12-65位氨基酸前后位置可以互换,并且可以被具有相同作用的其他序列替换;序列1中第12-65位氨基酸残基构成能够和β-arrestin2蛋白质相互作用的区域。
实施例2多肽TAT-AT1R-1的应用研究
心肌细胞缺氧/复氧模型:培养基中加入TAT-AT1R-1多肽(50μg/ml),对照组给与等体积生理盐水,预培养6h,以5%N2和95%CO2混合气体预饱和的无糖无血清培养基替换原培养基,转入5%N2和95%CO2的环境下缺氧处理12h,再换成含血清的高糖DMEM培养基在正常培养箱内继续孵育6h。台盼蓝染色计数死亡细胞数。
采用8-12周鼠龄的C57BL/6J小鼠,静脉给与TAT-AT1R-1多肽(1mg/kg),对照组给与等体积生理盐水。心肌缺血/再灌注损伤模型小鼠左冠状动脉前降支结扎30min局部缺血随后再灌注24h,再灌注结束后取材心脏组织,置于-80℃冰箱冰冻数分钟,心脏沿心尖至心底以横切面切成均匀厚度的环形切片,放置于盛有1%TTC染液的无菌器皿内,避光、恒温孵育后置于10%的甲醛内固定24h,检测心肌损伤面积。
结果显示:给予多肽后细胞生存状态和细胞形态未发现明显异常。图1为TAT-AT1R-1对心肌细胞缺氧/复氧保护作用检测,相对于对照组,加入TAT-AT1R-1多肽组AC16心肌细胞死亡率降低约52%;图2为TAT-AT1R-1对心肌缺血/再灌注损伤的保护作用检测,相对于对照组,注射TAT-AT1R-1多肽组心肌损伤面积减小约38.5%。如图1、图2综合表明,TAT-AT1R-1能够明显改善心肌缺血/再灌注对细胞的损伤。
综上所述:本发明所述多肽TAT-AT1R-1能够替代AT1R和β-arrestin2发生相互作用,从而阻断β-arrestin2信号通路对心肌缺血/再灌注损伤起到保护作用。
以上所述的具体实施方式,对本发明的目的、技术方案和有益效果进行了进一步详细说明,所应理解的是,以上所述仅为本发明的具体实施方式而已,并不用于限制本发明,凡在本发明的精神和原则之内,所做的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
序列表
<110> 徐州市中心医院
<120> 一种多肽、及其制备方法和应用
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 64
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg Gly Lys Lys Phe Lys
1 5 10 15
Arg Tyr Phe Leu Gln Leu Leu Lys Tyr Ile Pro Pro Lys Ala Lys Ser
20 25 30
His Ser Asn Leu Ser Thr Lys Ser Thr Leu Ser Tyr Arg Pro Ser Asp
35 40 45
Asn Val Ser Ser Ser Thr Lys Lys Pro Ala Pro Cys Phe Glu Val Glu
50 55 60
<210> 2
<211> 198
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
tacggtcgta aaaaacgtcg tcagcgtcgt cgtgggaaaa aatttaaaag atattttctc 60
cagcttctaa aatatattcc cccaaaagcc aaatcccact caaacctttc aacaaaaatg 120
agcacgcttt cctaccgccc ctcagataat gtaagctcat ccaccaagaa gcctgcacca 180
tgttttgagg ttgagtga 198
<210> 3
<211> 74
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
ccatctcgag atgtacggtc gtaaaaaacg tcgtcagcgt cgtcgtggga aaaaatttaa 60
aagatatttt ctcc 74
<210> 4
<211> 33
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
cgtcggatcc ctcaacctca aaacatggtg cag 33
Claims (4)
1.一种多肽或核酸分子在制备保护缺血/再灌注损伤作用的药物中的应用,其特征在于,所述多肽名称为TAT-AT1R-1,为如下(1)或(2)的多肽:
(1)SEQ ID NO:1所示的氨基酸序列第12-65位氨基酸残基;
(2)SEQ ID NO:1所示的氨基酸序列;
编码所述多肽的核酸分子,为如下(3)或(4)的DNA分子:
(3)SEQ ID NO:2自5’末端起第34位至第198位核苷酸所示的DNA分子;
(4)SEQ ID NO:2所示的DNA分子。
2.根据权利要求1所述一种多肽或核酸分子在制备保护缺血/再灌注损伤作用的药物中的应用,其特征在于,所述多肽的制备方法步骤为:
S1:构建表达载体,筛选重组子
提取AC16心肌细胞总RNA,反转录得到cDNA,以cDNA为模板通过PCR获得AT1R羧基末端多肽的核酸序列,在引物中插入TAT编码序列和酶切位点,酶切连接法构建表达载体pWaldo-TAT-AT1R-1,转化大肠杆菌BL21表达菌株,筛选重组子;
S2:获得高纯度的TAT-AT1R-1和GFP融合蛋白
接种重组子菌株于培养基中,诱导蛋白质表达,继续培养后离心收集细胞用于蛋白质纯化;将细胞悬浮于裂解缓冲液中,破碎细胞后去除细胞碎片;进一步纯化蛋白质可获得较高纯度的融合蛋白;
S3:纯化得到TAT-AT1R-1多肽
将S2得到的蛋白质样品中加入TEV蛋白酶,酶切过夜,去掉没有酶切完全的融合蛋白和酶切后的GFP,最终收集得到纯化的TAT-AT1R-1多肽样品,浓缩备用。
3.一种活性成分为权利要求1中所述的多肽或核酸分子的药品,具有保护心肌缺血/再灌注损伤作用。
4.根据权利要求3所述的药品,其特征在于,所述药品的给药方式包括但不限定:肌肉注射、静脉注射、心内注射、皮内注射、皮下注射、腹腔注射。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202111623326.6A CN114249837B (zh) | 2021-12-28 | 2021-12-28 | 一种多肽、及其制备方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202111623326.6A CN114249837B (zh) | 2021-12-28 | 2021-12-28 | 一种多肽、及其制备方法和应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN114249837A CN114249837A (zh) | 2022-03-29 |
CN114249837B true CN114249837B (zh) | 2023-10-20 |
Family
ID=80795422
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202111623326.6A Active CN114249837B (zh) | 2021-12-28 | 2021-12-28 | 一种多肽、及其制备方法和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114249837B (zh) |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101600715A (zh) * | 2006-12-21 | 2009-12-09 | 辉瑞产品公司 | 具有血管紧张素II受体拮抗作用和PPARγ活化活性的化合物 |
CN107929714A (zh) * | 2017-12-01 | 2018-04-20 | 广东医科大学 | 一种多肽在预防或治疗脑缺血再灌注损伤相关疾病中的应用 |
WO2018121457A1 (zh) * | 2016-12-29 | 2018-07-05 | 广东医科大学 | 一种多肽在预防或治疗心肌缺血再灌注损伤相关疾病中的药物应用 |
CN112225821A (zh) * | 2020-10-21 | 2021-01-15 | 徐州医科大学 | 一种具有抗肿瘤作用的多肽及其应用 |
-
2021
- 2021-12-28 CN CN202111623326.6A patent/CN114249837B/zh active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101600715A (zh) * | 2006-12-21 | 2009-12-09 | 辉瑞产品公司 | 具有血管紧张素II受体拮抗作用和PPARγ活化活性的化合物 |
WO2018121457A1 (zh) * | 2016-12-29 | 2018-07-05 | 广东医科大学 | 一种多肽在预防或治疗心肌缺血再灌注损伤相关疾病中的药物应用 |
CN107929714A (zh) * | 2017-12-01 | 2018-04-20 | 广东医科大学 | 一种多肽在预防或治疗脑缺血再灌注损伤相关疾病中的应用 |
CN112225821A (zh) * | 2020-10-21 | 2021-01-15 | 徐州医科大学 | 一种具有抗肿瘤作用的多肽及其应用 |
Non-Patent Citations (4)
Title |
---|
MicroRNA-214在心肌缺血/再灌注的研究进展;柳培雨等;现代生物医学进展;第16卷(第21期);4194-4196,4141 * |
Novel Subtype of Human Angiotensin II Type 1 Receptor: cDNA Cloning and Expression;Konishi H 等;Biochemical and biophysical Research Communications;第199卷(第2期);467-474 * |
The novel angiotensin II type 1 receptor (AT1R)-associated protein ATRAP downregulates AT1R and ameliorates cardiomyocyte hypertrophy;Yutaka Tanaka等;FEBS Letters;第579卷;1579–1586 * |
神经调节蛋白-1对心肌缺血再灌注损伤的保护作用;王富华等;生理科学进展;第47卷(第03期);223-226 * |
Also Published As
Publication number | Publication date |
---|---|
CN114249837A (zh) | 2022-03-29 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
AU2021218103B2 (en) | GDF15 fusion proteins and uses thereof | |
JP4411330B2 (ja) | 腫瘍壊死因子関連リガンド | |
JP3484189B2 (ja) | ヒトC3b/C4bレセプター (CR1) | |
US5856126A (en) | Peptide having anti-thrombus activity and method of producing the same | |
US11306296B2 (en) | MG53 mutants, methods of making the same, and uses thereof | |
JPH04281790A (ja) | Dnaおよびその用途 | |
WO1989009277A1 (en) | Mutant human angiogenin (angiogenesis factor with superior angiogenin activity) genes therefor and methods of expression | |
JP6230158B2 (ja) | ヒトβ−ヘキソサミニダーゼBの基質特異性を変換し、且つ、プロテアーゼ抵抗性を付与した新規高機能酵素 | |
CN107022557B (zh) | 新的死亡受体5抗体融合蛋白及其应用 | |
US5516656A (en) | Production of a new hirudin analog and anticoagulant pharmaceutical composition containing the same | |
NO302527B1 (no) | Fremgangsmåte ved fremstilling av polypeptid med antikoagulerende aktivitet | |
CN101134105B (zh) | 用于治疗和/或预防脑梗塞的含有重组人胰激肽原酶的药物组合物 | |
CA2565227A1 (en) | Human complement c3 derivates with cobra venom factor-like function | |
WO1998029446A1 (fr) | Neuropeptides provenant du scorpion | |
CN112225821B (zh) | 一种具有抗肿瘤作用的多肽及其应用 | |
CN114249837B (zh) | 一种多肽、及其制备方法和应用 | |
WO1998033067A1 (en) | Transcription factors that repress hiv transcription and methods based thereon | |
CA2130561A1 (en) | Modified tcf | |
CN107987144B (zh) | 一种蜈蚣多肽SLP_SsTx及其编码基因和应用 | |
WO2017063185A1 (zh) | 一种Slit2D2-HSA重组蛋白及其在治疗脓毒症中的应用 | |
CN114262384B (zh) | 一种多肽及其保护心肌缺血/再灌注损伤作用的应用 | |
KR20080026085A (ko) | 곤충세포에서 제조한 재조합 e-셀렉틴 | |
JP5982394B2 (ja) | MMP基質で連結したβig−h3断片ペプチド及びそのリウマチ性関節炎予防及び治療用途 | |
CN101062948B (zh) | 单体速效胰岛素及其制法和用途 | |
KR100724331B1 (ko) | 신규한 타키키닌 펩티드, 이의 전구체 펩티드 및 이를코딩하는 유전자 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |