CN113735939A - Combined polypeptide and application thereof - Google Patents
Combined polypeptide and application thereof Download PDFInfo
- Publication number
- CN113735939A CN113735939A CN202110873583.9A CN202110873583A CN113735939A CN 113735939 A CN113735939 A CN 113735939A CN 202110873583 A CN202110873583 A CN 202110873583A CN 113735939 A CN113735939 A CN 113735939A
- Authority
- CN
- China
- Prior art keywords
- arg
- lys
- ala
- pro
- gln
- 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.)
- Pending
Links
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 108
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 75
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 72
- CQOVPNPJLQNMDC-ZETCQYMHSA-N carnosine Chemical compound [NH3+]CCC(=O)N[C@H](C([O-])=O)CC1=CNC=N1 CQOVPNPJLQNMDC-ZETCQYMHSA-N 0.000 claims abstract description 55
- 108010087806 Carnosine Proteins 0.000 claims abstract description 51
- CQOVPNPJLQNMDC-UHFFFAOYSA-N N-beta-alanyl-L-histidine Natural products NCCC(=O)NC(C(O)=O)CC1=CN=CN1 CQOVPNPJLQNMDC-UHFFFAOYSA-N 0.000 claims abstract description 51
- QRYRORQUOLYVBU-VBKZILBWSA-N Carnosic acid Natural products CC([C@@H]1CC2)(C)CCC[C@]1(C(O)=O)C1=C2C=C(C(C)C)C(O)=C1O QRYRORQUOLYVBU-VBKZILBWSA-N 0.000 claims abstract description 50
- 229940044199 carnosine Drugs 0.000 claims abstract description 50
- 239000012634 fragment Substances 0.000 claims abstract description 31
- 108010085443 Anserine Proteins 0.000 claims abstract description 24
- SLRNWACWRVGMKD-UHFFFAOYSA-N L-anserine Natural products CN1C=NC(CC(NC(=O)CCN)C(O)=O)=C1 SLRNWACWRVGMKD-UHFFFAOYSA-N 0.000 claims abstract description 24
- 241000210053 Potentilla elegans Species 0.000 claims abstract description 24
- MYYIAHXIVFADCU-QMMMGPOBSA-N anserine Chemical compound CN1C=NC=C1C[C@H](NC(=O)CC[NH3+])C([O-])=O MYYIAHXIVFADCU-QMMMGPOBSA-N 0.000 claims abstract description 24
- 239000003814 drug Substances 0.000 claims abstract description 22
- CHFFHQUVXHEGBY-GARJFASQSA-N Ala-Lys-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N1CCC[C@@H]1C(=O)O)N CHFFHQUVXHEGBY-GARJFASQSA-N 0.000 claims abstract description 11
- IXHKPDJKKCUKHS-GARJFASQSA-N Lys-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCCCN)N IXHKPDJKKCUKHS-GARJFASQSA-N 0.000 claims abstract description 11
- FYQSMXKJYTZYRP-DCAQKATOSA-N Pro-Ala-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1 FYQSMXKJYTZYRP-DCAQKATOSA-N 0.000 claims abstract description 11
- 208000032382 Ischaemic stroke Diseases 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 6
- RVLOMLVNNBWRSR-KNIFDHDWSA-N (2s)-2-aminopropanoic acid;(2s)-2,6-diaminohexanoic acid Chemical group C[C@H](N)C(O)=O.NCCCC[C@H](N)C(O)=O RVLOMLVNNBWRSR-KNIFDHDWSA-N 0.000 claims description 5
- WPWUFUBLGADILS-WDSKDSINSA-N Ala-Pro Chemical group C[C@H](N)C(=O)N1CCC[C@H]1C(O)=O WPWUFUBLGADILS-WDSKDSINSA-N 0.000 claims description 5
- 108010065920 Insulin Lispro Chemical group 0.000 claims description 5
- QOOWRKBDDXQRHC-BQBZGAKWSA-N L-lysyl-L-alanine Chemical group OC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN QOOWRKBDDXQRHC-BQBZGAKWSA-N 0.000 claims description 5
- AIXUQKMMBQJZCU-IUCAKERBSA-N Lys-Pro Chemical group NCCCC[C@H](N)C(=O)N1CCC[C@H]1C(O)=O AIXUQKMMBQJZCU-IUCAKERBSA-N 0.000 claims description 5
- FELJDCNGZFDUNR-WDSKDSINSA-N Pro-Ala Chemical group OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1 FELJDCNGZFDUNR-WDSKDSINSA-N 0.000 claims description 5
- 108010087924 alanylproline Chemical group 0.000 claims description 5
- 210000004899 c-terminal region Anatomy 0.000 claims description 5
- 150000003839 salts Chemical class 0.000 claims description 5
- JOCMUACTOZLBLC-WOYTXXSLSA-N (2s)-6-amino-2-[[(2s)-2-[[(2s)-1-[(2s)-2-[[(2s)-2-aminopropanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]pyrrolidine-2-carbonyl]amino]propanoyl]amino]hexanoic acid Chemical group NC(N)=NCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O JOCMUACTOZLBLC-WOYTXXSLSA-N 0.000 claims description 3
- 208000024827 Alzheimer disease Diseases 0.000 claims description 3
- DBKNLHKEVPZVQC-LPEHRKFASA-N Arg-Ala-Pro Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@@H]1C(O)=O DBKNLHKEVPZVQC-LPEHRKFASA-N 0.000 claims description 3
- 208000016988 Hemorrhagic Stroke Diseases 0.000 claims description 3
- 208000018737 Parkinson disease Diseases 0.000 claims description 3
- 108010091735 fibrinogen peptide 6A Chemical group 0.000 claims description 3
- 208000020658 intracerebral hemorrhage Diseases 0.000 claims description 3
- 230000004770 neurodegeneration Effects 0.000 claims description 3
- 208000015122 neurodegenerative disease Diseases 0.000 claims description 3
- PIXQDIGKDNNOOV-GUBZILKMSA-N Ala-Lys-Gln Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O PIXQDIGKDNNOOV-GUBZILKMSA-N 0.000 claims description 2
- 108700000788 Human immunodeficiency virus 1 tat peptide (47-57) Proteins 0.000 claims description 2
- 208000030886 Traumatic Brain injury Diseases 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- GRRXPUAICOGISM-RWMBFGLXSA-N Arg-Lys-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCCN)NC(=O)[C@H](CCCN=C(N)N)N)C(=O)O GRRXPUAICOGISM-RWMBFGLXSA-N 0.000 claims 1
- -1 Gly-Arg-Pro-Ala-Lys Chemical compound 0.000 claims 1
- 241000270295 Serpentes Species 0.000 abstract description 14
- 230000000694 effects Effects 0.000 abstract description 9
- 208000012902 Nervous system disease Diseases 0.000 abstract description 5
- 108010038807 Oligopeptides Proteins 0.000 abstract description 4
- 102000015636 Oligopeptides Human genes 0.000 abstract description 4
- 230000004071 biological effect Effects 0.000 abstract description 4
- 230000008499 blood brain barrier function Effects 0.000 abstract description 3
- 210000001218 blood-brain barrier Anatomy 0.000 abstract description 3
- 230000002949 hemolytic effect Effects 0.000 abstract description 3
- 238000001990 intravenous administration Methods 0.000 abstract description 3
- 208000025966 Neurological disease Diseases 0.000 abstract description 2
- 208000029028 brain injury Diseases 0.000 abstract description 2
- 230000015556 catabolic process Effects 0.000 abstract description 2
- 238000006731 degradation reaction Methods 0.000 abstract description 2
- 239000002207 metabolite Substances 0.000 abstract description 2
- 102000009123 Fibrin Human genes 0.000 abstract 1
- 108010073385 Fibrin Proteins 0.000 abstract 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 abstract 1
- 229950003499 fibrin Drugs 0.000 abstract 1
- 230000000149 penetrating effect Effects 0.000 abstract 1
- 238000005336 cracking Methods 0.000 description 49
- 239000011347 resin Substances 0.000 description 41
- 229920005989 resin Polymers 0.000 description 41
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 30
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 24
- 241000700159 Rattus Species 0.000 description 22
- 230000015572 biosynthetic process Effects 0.000 description 21
- 239000003153 chemical reaction reagent Substances 0.000 description 21
- 238000003786 synthesis reaction Methods 0.000 description 21
- BZLVMXJERCGZMT-UHFFFAOYSA-N Methyl tert-butyl ether Chemical compound COC(C)(C)C BZLVMXJERCGZMT-UHFFFAOYSA-N 0.000 description 20
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 20
- RDOXTESZEPMUJZ-UHFFFAOYSA-N anisole Chemical compound COC1=CC=CC=C1 RDOXTESZEPMUJZ-UHFFFAOYSA-N 0.000 description 20
- 230000008878 coupling Effects 0.000 description 20
- 238000010168 coupling process Methods 0.000 description 20
- 238000005859 coupling reaction Methods 0.000 description 20
- 238000004108 freeze drying Methods 0.000 description 20
- HNICLNKVURBTKV-NDEPHWFRSA-N (2s)-5-[[amino-[(2,2,4,6,7-pentamethyl-3h-1-benzofuran-5-yl)sulfonylamino]methylidene]amino]-2-(9h-fluoren-9-ylmethoxycarbonylamino)pentanoic acid Chemical compound C12=CC=CC=C2C2=CC=CC=C2C1COC(=O)N[C@H](C(O)=O)CCCN=C(N)NS(=O)(=O)C1=C(C)C(C)=C2OC(C)(C)CC2=C1C HNICLNKVURBTKV-NDEPHWFRSA-N 0.000 description 16
- 125000006239 protecting group Chemical group 0.000 description 14
- 150000001413 amino acids Chemical group 0.000 description 13
- 241001465754 Metazoa Species 0.000 description 12
- 229940024606 amino acid Drugs 0.000 description 12
- 210000005013 brain tissue Anatomy 0.000 description 12
- 229940079593 drug Drugs 0.000 description 12
- 239000000243 solution Substances 0.000 description 12
- 238000005406 washing Methods 0.000 description 12
- 239000012190 activator Substances 0.000 description 11
- 210000004004 carotid artery internal Anatomy 0.000 description 11
- 239000000843 powder Substances 0.000 description 11
- JFLSOKIMYBSASW-UHFFFAOYSA-N 1-chloro-2-[chloro(diphenyl)methyl]benzene Chemical compound ClC1=CC=CC=C1C(Cl)(C=1C=CC=CC=1)C1=CC=CC=C1 JFLSOKIMYBSASW-UHFFFAOYSA-N 0.000 description 10
- 230000003213 activating effect Effects 0.000 description 10
- 239000003795 chemical substances by application Substances 0.000 description 10
- 239000011248 coating agent Substances 0.000 description 10
- 238000000576 coating method Methods 0.000 description 10
- 239000012043 crude product Substances 0.000 description 10
- 238000007689 inspection Methods 0.000 description 10
- UZKWTJUDCOPSNM-UHFFFAOYSA-N methoxybenzene Substances CCCCOC=C UZKWTJUDCOPSNM-UHFFFAOYSA-N 0.000 description 10
- 239000002244 precipitate Substances 0.000 description 10
- 230000001376 precipitating effect Effects 0.000 description 10
- 238000002953 preparative HPLC Methods 0.000 description 10
- 238000000746 purification Methods 0.000 description 10
- 238000012546 transfer Methods 0.000 description 10
- 238000005303 weighing Methods 0.000 description 10
- 210000000269 carotid artery external Anatomy 0.000 description 8
- 230000002490 cerebral effect Effects 0.000 description 8
- XWQVQFBTSBCKLI-FKXNDIMNSA-N dnc008987 Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCNC(N)=N)NC(=O)CNC(=O)[C@@H](N)CC1=CC=C(O)C=C1 XWQVQFBTSBCKLI-FKXNDIMNSA-N 0.000 description 8
- 108010016626 Dipeptides Proteins 0.000 description 7
- 206010061216 Infarction Diseases 0.000 description 7
- 230000007574 infarction Effects 0.000 description 7
- 239000007790 solid phase Substances 0.000 description 7
- 210000001367 artery Anatomy 0.000 description 6
- UCMIRNVEIXFBKS-UHFFFAOYSA-N beta-alanine Chemical compound NCCC(O)=O UCMIRNVEIXFBKS-UHFFFAOYSA-N 0.000 description 6
- 206010008118 cerebral infarction Diseases 0.000 description 6
- 238000002347 injection Methods 0.000 description 6
- 239000007924 injection Substances 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- QWXZOFZKSQXPDC-NSHDSACASA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)propanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](C)C(O)=O)C3=CC=CC=C3C2=C1 QWXZOFZKSQXPDC-NSHDSACASA-N 0.000 description 5
- 108700019745 Disks Large Homolog 4 Proteins 0.000 description 5
- 102000047174 Disks Large Homolog 4 Human genes 0.000 description 5
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 5
- 210000001168 carotid artery common Anatomy 0.000 description 5
- 230000006870 function Effects 0.000 description 5
- 210000003462 vein Anatomy 0.000 description 5
- UMRUUWFGLGNQLI-QFIPXVFZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-6-[(2-methylpropan-2-yl)oxycarbonylamino]hexanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCCCNC(=O)OC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 UMRUUWFGLGNQLI-QFIPXVFZSA-N 0.000 description 4
- 201000006474 Brain Ischemia Diseases 0.000 description 4
- 206010008120 Cerebral ischaemia Diseases 0.000 description 4
- 210000004556 brain Anatomy 0.000 description 4
- 229960002885 histidine Drugs 0.000 description 4
- 230000003834 intracellular effect Effects 0.000 description 4
- 230000000302 ischemic effect Effects 0.000 description 4
- 229940121466 nerinetide Drugs 0.000 description 4
- 238000004393 prognosis Methods 0.000 description 4
- XXMYDXUIZKNHDT-QNGWXLTQSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-(1-tritylimidazol-4-yl)propanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)C(N=C1)=CN1C(C=1C=CC=CC=1)(C=1C=CC=CC=1)C1=CC=CC=C1 XXMYDXUIZKNHDT-QNGWXLTQSA-N 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 239000003146 anticoagulant agent Substances 0.000 description 3
- 239000003963 antioxidant agent Substances 0.000 description 3
- 230000003078 antioxidant effect Effects 0.000 description 3
- 230000003542 behavioural effect Effects 0.000 description 3
- 229940000635 beta-alanine Drugs 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 3
- 230000000324 neuroprotective effect Effects 0.000 description 3
- 238000005502 peroxidation Methods 0.000 description 3
- 239000000902 placebo Substances 0.000 description 3
- 229940068196 placebo Drugs 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 230000002537 thrombolytic effect Effects 0.000 description 3
- ZPGDWQNBZYOZTI-SFHVURJKSA-N (2s)-1-(9h-fluoren-9-ylmethoxycarbonyl)pyrrolidine-2-carboxylic acid Chemical compound OC(=O)[C@@H]1CCCN1C(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21 ZPGDWQNBZYOZTI-SFHVURJKSA-N 0.000 description 2
- 206010008190 Cerebrovascular accident Diseases 0.000 description 2
- 102100022397 Nitric oxide synthase, brain Human genes 0.000 description 2
- 101710111444 Nitric oxide synthase, brain Proteins 0.000 description 2
- 208000006011 Stroke Diseases 0.000 description 2
- 102000003978 Tissue Plasminogen Activator Human genes 0.000 description 2
- 108090000373 Tissue Plasminogen Activator Proteins 0.000 description 2
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 description 2
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 229960003318 alteplase Drugs 0.000 description 2
- 230000036760 body temperature Effects 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 208000026106 cerebrovascular disease Diseases 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 210000003205 muscle Anatomy 0.000 description 2
- 239000004090 neuroprotective agent Substances 0.000 description 2
- 150000002825 nitriles Chemical class 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 230000010410 reperfusion Effects 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 239000012192 staining solution Substances 0.000 description 2
- 230000035882 stress Effects 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 229940116269 uric acid Drugs 0.000 description 2
- XZWXFWBHYRFLEF-FSPLSTOPSA-N Ala-His Chemical compound C[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 XZWXFWBHYRFLEF-FSPLSTOPSA-N 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 206010008092 Cerebral artery thrombosis Diseases 0.000 description 1
- 241000283153 Cetacea Species 0.000 description 1
- 208000017667 Chronic Disease Diseases 0.000 description 1
- 235000009161 Espostoa lanata Nutrition 0.000 description 1
- 240000001624 Espostoa lanata Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 102100022630 Glutamate receptor ionotropic, NMDA 2B Human genes 0.000 description 1
- 101710195187 Glutamate receptor ionotropic, NMDA 2B Proteins 0.000 description 1
- 241000282553 Macaca Species 0.000 description 1
- JDHILDINMRGULE-LURJTMIESA-N N(pros)-methyl-L-histidine Chemical compound CN1C=NC=C1C[C@H](N)C(O)=O JDHILDINMRGULE-LURJTMIESA-N 0.000 description 1
- BRMWTNUJHUMWMS-LURJTMIESA-N N(tele)-methyl-L-histidine Chemical compound CN1C=NC(C[C@H](N)C(O)=O)=C1 BRMWTNUJHUMWMS-LURJTMIESA-N 0.000 description 1
- 102000004868 N-Methyl-D-Aspartate Receptors Human genes 0.000 description 1
- 108090001041 N-Methyl-D-Aspartate Receptors Proteins 0.000 description 1
- 108090000189 Neuropeptides Proteins 0.000 description 1
- 102000003797 Neuropeptides Human genes 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 206010048038 Wound infection Diseases 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 238000009098 adjuvant therapy Methods 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 108010070944 alanylhistidine Proteins 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- RNFNDJAIBTYOQL-UHFFFAOYSA-N chloral hydrate Chemical compound OC(O)C(Cl)(Cl)Cl RNFNDJAIBTYOQL-UHFFFAOYSA-N 0.000 description 1
- 229960002327 chloral hydrate Drugs 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000002224 dissection Methods 0.000 description 1
- 101150069842 dlg4 gene Proteins 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 229940124645 emergency medicine Drugs 0.000 description 1
- 238000005516 engineering process Methods 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
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 125000002485 formyl group Chemical class [H]C(*)=O 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 208000028867 ischemia Diseases 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 210000003657 middle cerebral artery Anatomy 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000000626 neurodegenerative effect Effects 0.000 description 1
- 230000000926 neurological effect Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000025308 nuclear transport Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 239000008177 pharmaceutical agent Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000013641 positive control Substances 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
- 230000001737 promoting effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000009993 protective function Effects 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000011552 rat model Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000010532 solid phase synthesis reaction Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 210000001913 submandibular gland Anatomy 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000013151 thrombectomy Methods 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
- 210000001186 vagus nerve Anatomy 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
- A61P25/16—Anti-Parkinson drugs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- 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
- A61P9/12—Antihypertensives
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Neurosurgery (AREA)
- Neurology (AREA)
- Biomedical Technology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Heart & Thoracic Surgery (AREA)
- Cardiology (AREA)
- Psychology (AREA)
- Hospice & Palliative Care (AREA)
- Psychiatry (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明公开了一种组合多肽及其应用，属于药物技术领域。该组合肽由多肽片段A和多肽片段B组合而成，多肽片段A为TAT穿膜肽，多肽片段B为由Pro‑Ala‑Lys、Ala‑Lys‑Pro、Lys‑Ala‑Pro及其片段和衍生物中的一种或多种组成且其C端和/或N端具有肌肽、鹅肌肽或者蛇肌肽。多肽片段B与溶血活性寡肽纤维蛋白beta链的降解或代谢产物（P6A）相关，多肽片段A对多肽片段B具有提升作用，采用TAT穿膜肽对现有多肽进行进一步提升。这些肽类出乎意外地容易透过血脑屏障，通过静脉途径给药展示了很好的生物活性，从而在治疗神经系统疾病，尤其是脑损伤，脑卒中方面显示了广阔的前景。The invention discloses a combined polypeptide and application thereof, belonging to the technical field of medicine. The combined peptide is composed of a polypeptide fragment A and a polypeptide fragment B, wherein the polypeptide fragment A is a TAT penetrating peptide, and the polypeptide fragment B is composed of Pro-Ala-Lys, Ala-Lys-Pro, Lys-Ala-Pro and its fragments and One or more of the derivatives consists of carnosine, anserine or snake carnosine at its C-terminus and/or N-terminus. Polypeptide fragment B is related to the degradation or metabolite (P6A) of the hemolytic active oligopeptide fibrin beta chain. Polypeptide fragment A has an improving effect on polypeptide fragment B, and TAT transmembrane peptide is used to further improve the existing polypeptide. These peptides are unexpectedly easy to pass through the blood-brain barrier, and show good biological activity by intravenous administration, thus showing broad prospects in the treatment of neurological diseases, especially brain injury and stroke.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a combined polypeptide and application thereof.
Background
Nerinetide (NA-1) is a neuroprotective agent that interferes with postsynaptic density protein 95 (PSD-95) by terminating the production of intracellular NO free radicals, reduces infarct size of cerebral ischemia reperfusion in preclinical ischemic stroke models, and improves functional prognosis. Patients with acute ischemic stroke with macrovascular occlusion within the adult 12-hour treatment window were randomized to receive a single dose of 2.6mg/kg of Nerinetide and a maximum dose of 270 mg or saline placebo. The primary endpoint of the study was a good functional prognosis after 90 days of randomization, defined as a Modified Rankin Scale (MRS) score of 0-2, and the secondary endpoint was neurological disability, functional independence in daily living activities, good functional outcome (MRS 0-1), and mortality. There were a total of 1105 patients enrolled in the experiment, of which Nerinetide group 549 human and placebo group 556 human. The proportion of patients with an mRS score of 0-2 over 90 days was: nerinetide group 337 (61.4%), placebo group 329 (59.2%). Secondary results were similar between the two groups. At the same time, the present study found that netitenetide treatment resulted in an inhibition of the alteplase effect in patients receiving alteplase. The incidence of serious adverse events was the same between the two groups. (Michael D Hill et al, effectiveness and safety of a nitrile for the treatment of an acid electrochemical stress (ESCAPE-NA 1): a multicentre, double-blind, randomised controlled three. Lance. 2020, 395 (10227), P878-887).
The Nerinetide is a fusion peptide consisting of 9C-terminal residues of the NMDAR GluN2B subunit and a transmembrane peptide TAT derived from nuclear transport activator, can be combined with PDZ-1 or PDZ-2 structural domain of PSD95 to inhibit the generation of nNOS NO, and is also called TAT-NR2B9C, and the specific amino acid sequence thereof is as follows: Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Leu-Ser-Ser-Ile-Glu-Ser-Asp-Val. The latter Lys-Leu-Ser-Ser-Ile-Glu-Ser-Asp-Val sequence is NR2B9c sequence, and specifically inhibits the generation of nNOS NO. The former part Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg is to improve the bioavailability of NR2B9 c.
Carnosine (L-Carnosine), a dipeptide consisting of two amino acids, beta-alanine and L-histidine. Carnosine has strong antioxidant capacity and is beneficial to human body. Carnosine has been shown to scavenge reactive oxygen Radicals (ROS) and α - β unsaturated aldehydes, which form during oxidative stress over-oxidation of cell membrane fatty acids. Carnosine has anti-inflammatory, anti-glycation, anti-oxidation and chelating effects, can be used as an over-the-counter food supplement, and has good prospects in the aspects of prevention and adjuvant therapy of cardiovascular diseases, neurodegenerative and other chronic diseases. In animal experiments, the neuroprotective mechanism of carnosine can prevent permanent cerebral ischemia. Carnosine is also an important intracellular antioxidant. Carnosine is not only non-toxic but also has strong antioxidant properties, and thus has attracted much attention as a novel food additive and pharmaceutical agent. Carnosine is involved in intracellular peroxidation, and can inhibit related peroxidation in cells in addition to inhibiting peroxidation of cell membranes. In 1900, the russian scholars first discovered carnosine, and later, the russian scholars extracted and separated other histidine dipeptide derivatives in different muscle tissues, such as Anserine (Anserine), which is a dipeptide consisting of two amino acids, beta-alanine and 1-methyl-L-histidine, and whale carnosine or snake carnosine (also called opine), which is a dipeptide consisting of beta-alanine and 3-methyl-L-histidine. The content of the carnosine and the carnosine derivative is different among different animals, and the content and the proportion of the histidine dipeptides are different among different species, so that the carnosine and the carnosine derivative have certain specificity. In addition to histidine dipeptides being present in muscle tissue, these dipeptides are also present in other tissues, such as brain tissue. The carnosine derivatives have water solubility and strong activity, have remarkable functions of resisting oxidation, resisting aging, reducing uric acid and the like, are used as natural antioxidants and uric acid reduction dietary therapy in the food industry, and also have certain neuroprotective function.
Although carnosine and carnosine derivatives have some brain protective effects, they often require larger doses and are not as effective as carnosine, anserine and snake carnosine alone. The patent of 'a synthetic polypeptide and application thereof' (Lijiangxiong, Chinese patent application No. 202110266153.0) reported by the inventor in 2021 reports a brand-new polypeptide containing fragments of carnosine and the like, and the novel combined peptide provided by the invention also has high biological activity.
Disclosure of Invention
The invention provides a combined polypeptide, which is formed by linking oligopeptide with thrombolytic function and carnosine, anserine or snake carnosine and the like, and forms a series of polypeptide compounds with the characteristics of protective function of the carnosine, the anserine or the snake carnosine and thrombolytic function of the oligopeptide with thrombolytic function. The polypeptide fragment B (such as Ala-Arg-Pro-Ala-Lys, Arg-Pro-Ala-Lys and the like) is related to the degradation or metabolite (P6A) of the fibrinoprotein beta chain of hemolytic active oligopeptide, the polypeptide fragment A has a promoting effect on the polypeptide fragment B, the TAT transmembrane peptide Tyr-Gly-Arg-Lys-Arg-Arg-Gln-Arg-Arg-Arg is adopted to further improve the polypeptide with the application number of CN202110266153.0, and the polypeptide fragment B also has the bioactivity of the work, and the activity is further promoted according to the number of single molecules. The peptides unexpectedly and easily permeate blood brain barrier, and show good biological activity by intravenous administration, thereby showing wide prospect in the aspect of treating nervous system diseases, particularly brain injury and cerebral apoplexy. The technical scheme is as follows:
in one aspect, the present invention provides a combination polypeptide comprising a polypeptide fragment A and a polypeptide fragment B, wherein the polypeptide fragment A is Tyr-Gly-Arg-Lys-Arg-Arg-Gln-Arg-Arg-Arg, and the polypeptide fragment B is Pro-Ala-Lys, Ala-Lys-Pro, Lys-Ala-Pro, Pro-Ala, Ala-Lys, Lys-Pro, Lys-Ala, Ala-Pro, Ala-Arg-Pro-Ala-Lys, Ala-Arg-Ala-Lys-Pro, Ala-Lys-Ala-Pro, Arg-Ala-Lys-Pro, Arg-Lys-Ala-Pro, Gly-Arg-Pro-Ala-Lys, Arg-Ala-Pro, One or more of Gly-Arg-Ala-Lys-Pro, Gly-Arg-Lys-Ala-Pro, Gln-Arg-Pro-Ala-Lys, Gln-Arg-Ala-Lys-Pro, and Gln-Arg-Lys-Ala-Pro, and the like, and the C terminal and/or N terminal of the peptide has carnosine, anserine, or serosine.
Wherein the polypeptide fragment B consists of one or more of Pro-Ala-Lys, Ala-Lys-Pro, Lys-Ala-Pro, Pro-Ala, Ala-Lys, Lys-Pro, Lys-Ala and Ala-Pro, and the C end and/or N end of the polypeptide fragment B is provided with carnosine, anserine or serosine; the polypeptide fragment B preferably consists of one Pro-Ala-Lys, Ala-Lys-Pro or Lys-Ala-Pro and has carnosine, anserine or snake carnosine at the C-terminal and/or N-terminal; the polypeptide fragment B is preferably pentapeptide composed of one of Pro-Ala-Lys, Ala-Lys-Pro and Lys-Ala-Pro and one of Pro-Ala, Ala-Lys, Lys-Pro, Lys-Ala and Ala-Pro, and has carnosine, anserine or serosine at C-terminal and/or N-terminal. The method comprises the following steps:
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Pro-Ala，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Pro-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Lys-Ala，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Pro-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Pro-Ala，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Pro-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Lys-Ala，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Pro-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Pro-Ala，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Pro-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Lys-Ala，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Pro-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Pro-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Pro-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Lys-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Pro-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Pro-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Pro-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Lys-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Pro-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Pro-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Pro-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Lys-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Pro-Lys-Ala-Pro-β-Ala-His，
β-Ala-His-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Pro-Ala-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Pro-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Lys-Ala-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Pro-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Pro-Ala-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Pro-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Lys-Ala-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Pro-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Pro-Ala-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Pro-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Lys-Ala-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Pro-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Pro-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Pro-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Lys-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Pro-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Pro-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Pro-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Lys-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Pro-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Pro-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Ala-Pro-β-Ala-Hi-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Args，
Lys-Ala-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Pro-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Lys-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Pro-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
wherein His is His, 1-Methyl-His or 3-Methyl-His corresponding to carnosine, anserine and snake carnosine respectively.
Wherein the polypeptide fragment B is hexapeptide consisting of one or two of Pro-Ala-Lys, Ala-Lys-Pro and Lys-Ala-Pro (consisting of two Pro-Ala-Lys, two Ala-Lys-Pro or two Lys-Ala-Pro) and has carnosine, anserine or serosine at the C end and/or N end, and comprises:
Pro-Ala-Lys-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His -Pro-Ala-Lys-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His -Lys-Ala-Pro-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His -Lys-Ala-Pro-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Lys-Ala-Pro，
β-Ala-His-Pro-Ala-Lys-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Lys-Ala-Pro-β-Ala-His，
wherein His is His, 1-Methyl-His or 3-Methyl-His corresponding to carnosine, anserine and snake carnosine respectively.
Among them, the combined polypeptide of the present invention includes:
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-β-Ala-His，
β-Ala-His-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
wherein His is His, 1-Methyl-His or 3-Methyl-His corresponding to carnosine, anserine and snake carnosine respectively.
Specifically, the combined polypeptide of the present invention includes (the polypeptide fragment B is Pro-Ala-Lys and has carnosine, anserine, or serosine at the C-terminus and/or N-terminus):
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-β-Ala-His，
β-Ala-His-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
wherein His is His, 1-Methyl-His or 3-Methyl-His corresponding to carnosine, anserine and snake carnosine respectively.
Specifically, the combined polypeptide of the invention comprises (the polypeptide fragment B is Ala-Lys-Pro and has carnosine, anserine or snake carnosine at the C terminal and/or N terminal):
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-β-Ala-His，
β-Ala-His-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
wherein His is His, 1-Methyl-His or 3-Methyl-His corresponding to carnosine, anserine and snake carnosine respectively.
Specifically, the combined polypeptide of the present invention includes (the polypeptide fragment B is Lys-Ala-Pro and has carnosine, anserine, or serosine at the C-terminus and/or N-terminus):
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-β-Ala-His，
β-Ala-His-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
wherein His is His, 1-Methyl-His or 3-Methyl-His corresponding to carnosine, anserine and snake carnosine respectively.
Preferably, the combined polypeptide of the invention comprises:
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-β-Ala-His；
wherein His is only His.
In another aspect, the invention also provides the application of the combined polypeptide and the salt thereof in preparing medicaments (especially medicaments for treating nervous system diseases).
Specifically, the combined polypeptide and the salt thereof are applied to the preparation of medicines for treating ischemic stroke, hemorrhagic stroke, cerebral trauma, Alzheimer disease, Parkinson disease or other neurodegenerative diseases and the like. Preferably, the combined polypeptide is applied to preparing a medicine for treating cerebral arterial thrombosis.
Among them, the above-mentioned drugs are injections, oral administration, sublingual administration, spray administration, anal administration, etc., and injections are preferred. Specifically, the injection is powder injection or injection. Further, the aforementioned drugs are administered intravenously. The medicine can also be used as an active ingredient to prepare other dosage forms so as to facilitate corresponding medical application. The developed preparation can be taken as neuroprotective medicine, oral administration and sublingual administration preparation, and can be taken as emergency medicine for cerebral apoplexy, and the developed preparation can be taken as spray administration, anal administration and other preparations, so that the patient without mobility can be treated.
Further, the medicament comprises a pharmaceutically acceptable diluent or/and a carrier and the like.
In yet another aspect, the invention provides methods for preparing various classes of combinatorial polypeptides of the invention by solid phase synthesis, but some of the peptides are more conveniently synthesized in liquid phase. The salification of polypeptide drugs is one of the common means for improving the physicochemical properties of drug molecules and improving the pharmaceutical properties of the drugs, and the drugs can be salts in any form.
In order to determine the application of the synthetic peptide in the aspect of nervous system diseases, SD rats are used as experimental objects, a cerebral ischemia rat model is prepared by adopting a cerebral middle artery occlusion Method (MCAO), a medicine is injected intravenously after 1-2 hours of ischemia, and behavioral observation and scoring are carried out on each animal at 22-24 hours. After the behavioral observation, the experimental rats were euthanized and their brains were removed, sectioned in brain tissue, stained with TTC and then analyzed quantitatively to calculate the% cerebral infarction volume.
The combined peptide creatively combines hemolytic peptide, carnosine, anserine or snake carnosine and the like, and combines with TAT peptide, so that the biological activity is improved, and the blood brain barrier is improved. The synthetic peptide is preferably applied to treating ischemic stroke, hemorrhagic stroke, brain trauma, Alzheimer disease, Parkinson disease and other neurodegenerative diseases. The invention only needs 3mg/kg of synthetic polypeptide intravenous injection dosage and can achieve remarkable treatment effect.
Drawings
FIG. 1 is a section view of brain tissue of a model set;
FIG. 2 is a sectional view of brain tissue of group S1;
FIG. 3 is a section of brain tissue from group S2;
FIG. 4 is a section of brain tissue from group S4;
FIG. 5 is a section of brain tissue from group S5;
FIG. 6 is a section of brain tissue from group S6;
FIG. 7 is a sectional view of brain tissue from a sham-operated group.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail with reference to the accompanying drawings.
Example 1: synthesis of Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg- β -Ala-His-Lys-Ala-Pro:
1. Fmoc-Pro-CTC resin was obtained by coupling solid support 2-CTC resin and Fmoc-Pro-OH in the presence of an activator system (HoBT, DIC).
2. And (3) removing the Fmoc protecting group on the Fmoc-Pro-CTC by using 20% piperidine, and washing the Fmoc protecting group by using DMF after the Fmoc protecting group is completely removed.
3. Weighing 3-time excess Fmoc-Ala-OH, adding a small amount of DMF with three-time excess activating agent, fully dissolving, adding into the washed resin after dissolving, reacting for 1h, and washing with DMF.
4. And repeating the second step and the third step, and coupling the amino acids with the N-terminal Fmoc protection and the side chain protection in sequence from Lys to Tyr according to the main chain sequence.
5. After the synthesis is finished, the cracking is carried out, and the proportion of a cracking reagent is TFA: EDT (electro-thermal transfer coating): and Tis: TA: anisole: h2O = 80: 5: 1: 5: 5: 4 (volume ratio), 1g of peptide resin needs 10ml of cracking reagent, cracking for about 2h (120 r/min) at room temperature, precipitating with glacial methyl tert-butyl ether after cracking, and obtaining crude product as lower precipitate.
6. The crude peptide from the previous step was dissolved and purified on preparative HPLC with 0.1% TFA/acetonitrile.
7. And (4) freeze-drying after purification, taking out powder after freeze-drying, and performing subpackage quality inspection to obtain the target polypeptide with the purity of more than 95%.
Example 2: synthesis of Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg- β -Ala-His-Ala-Lys-Pro:
1. Fmoc-Pro-CTC resin was obtained by coupling solid support 2-CTC resin and Fmoc-Pro-OH in the presence of an activator system (HoBT, DIC).
2. And (3) removing the Fmoc protecting group on the Fmoc-Pro-CTC by using 20% piperidine, and washing the Fmoc protecting group by using DMF after the Fmoc protecting group is completely removed.
3. Weighing 3 times excess Fmoc-Lys (Boc) -OH and three times excess activating agent, adding a small amount of DMF to fully dissolve, adding the solution into a clean resin after the solution is completely dissolved, and washing the resin with DMF after reacting for 1 h.
4. And repeating the second step and the third step, and coupling the amino acids with the Fmoc protection at the N end and the protected side chains in sequence from Ala to Tyr according to the main chain sequence.
5. After the synthesis is finished, the cracking is carried out, and the proportion of a cracking reagent is TFA: EDT (electro-thermal transfer coating): and Tis: TA: anisole: h2O = 80: 5: 1: 5: 5: 4 (volume ratio), 1g of peptide resin needs 10ml of cracking reagent, cracking for about 2h (120 r/min) at room temperature, precipitating with glacial methyl tert-butyl ether after cracking, and obtaining crude product as lower precipitate.
6. The crude peptide from the previous step was dissolved and purified on preparative HPLC with 0.1% TFA/acetonitrile.
7. And (4) freeze-drying after purification, taking out powder after freeze-drying, and performing subpackage quality inspection to obtain the target polypeptide with the purity of more than 95%.
Example 3: synthesis of Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg- β -Ala-His-Pro-Ala-Lys:
1. Fmoc-Lys (Boc) -CTC resin was obtained by coupling solid support 2-CTC resin with Fmoc-Lys (Boc) -OH in the presence of an activator system (HoBT, DIC).
2. The Fmoc protecting group on Fmoc-Lys (Boc) -CTC was removed with 20% piperidine and washed clean with DMF.
3. Weighing 3-time excess Fmoc-Ala-OH, adding a small amount of DMF into three-time excess activating agent, fully dissolving, adding into the washed resin after dissolving, and washing with DMF after reaction.
4. And repeating the second step and the third step, and coupling the amino acids with the N-terminal Fmoc protection and the side chain protection in sequence from Pro to Tyr according to the main chain sequence.
5. After the synthesis is finished, the cracking is carried out, and the proportion of a cracking reagent is TFA: EDT (electro-thermal transfer coating): and Tis: TA: anisole: h2O = 80: 5: 1: 5: 5: 4 (volume ratio), 1g of peptide resin needs 10ml of cracking reagent, cracking for about 2h (120 r/min) at room temperature, precipitating with glacial methyl tert-butyl ether after cracking, and obtaining crude product as lower precipitate.
6. The crude peptide from the previous step was dissolved and purified on preparative HPLC with 0.1% TFA/acetonitrile.
7. And (4) freeze-drying after purification, taking out powder after freeze-drying, and performing subpackage quality inspection to obtain the target polypeptide with the purity of more than 95%.
Example 4: synthesis of Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys- β -Ala-His:
1. Fmoc-His (Trt) -CTC resin was obtained by coupling solid phase support 2-CTC resin with Fmoc-His (Trt) -OH in the presence of an activator system (HoBT, DIC).
2. The Fmoc protecting group on Fmoc-His (Trt) -CTC was removed with 20% piperidine, and washed clean with DMF.
3. Weighing 3-time excess Fmoc-Ala-OH, adding a small amount of DMF with three-time excess activating agent, fully dissolving, adding into the washed resin after dissolving, reacting for 1h, and washing with DMF.
4. And repeating the second step and the third step, and coupling the amino acids with the N-terminal Fmoc protection and the side chain protection in sequence from Lys to Tyr according to the main chain sequence.
5. After the synthesis is finished, the cracking is carried out, and the proportion of a cracking reagent is TFA: EDT (electro-thermal transfer coating): and Tis: TA: anisole: h2O = 80: 5: 1: 5: 5: 4 (volume ratio), 1g of peptide resin needs 10ml of cracking reagent, cracking for about 2h (120 r/min) at room temperature, precipitating with glacial methyl tert-butyl ether after cracking, and obtaining crude product as lower precipitate.
6. The crude peptide from the previous step was dissolved and purified on preparative HPLC with 0.1% TFA/acetonitrile.
7. And (4) freeze-drying after purification, taking out powder after freeze-drying, and subpackaging for quality inspection.
Example 5: synthesis of Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro- β -Ala-His:
1. Fmoc-His (Trt) -CTC resin was obtained by coupling solid phase support 2-CTC resin with Fmoc-His (Trt) -OH in the presence of an activator system (HoBT, DIC).
2. The Fmoc protecting group on Fmoc-His (Trt) -CTC was removed with 20% piperidine, and washed clean with DMF.
3. Weighing 3-time excess Fmoc-Ala-OH, adding a small amount of DMF with three-time excess activating agent, fully dissolving, adding into the washed resin after dissolving, reacting for 1h, and washing with DMF.
4. And repeating the second step and the third step, and coupling the amino acids with the N-terminal Fmoc protection and the side chain protection in sequence from Lys to Tyr according to the main chain sequence.
5. After the synthesis is finished, the cracking is carried out, and the proportion of a cracking reagent is TFA: EDT (electro-thermal transfer coating): and Tis: TA: anisole: h2O = 80: 5: 1: 5: 5: 4 (volume ratio), prepared by mixing 1g peptide resin with 10ml cracking reagent, cracking at room temperature for about 2h (120 r/min), and then using iceAnd (3) precipitating methyl tert-butyl ether, wherein the lower precipitate is a crude product.
6. The crude peptide from the previous step was dissolved and purified on preparative HPLC with 0.1% TFA/acetonitrile.
7. And (4) freeze-drying after purification, taking out powder after freeze-drying, and subpackaging for quality inspection.
Example 6: synthesis of Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro- β -Ala-His:
1. Fmoc-His (Trt) -CTC resin was obtained by coupling solid phase support 2-CTC resin with Fmoc-His (Trt) -OH in the presence of an activator system (HoBT, DIC).
2. The Fmoc protecting group on Fmoc-His (Trt) -CTC was removed with 20% piperidine, and washed clean with DMF.
3. Weighing 3-time excess Fmoc-Ala-OH, adding a small amount of DMF with three-time excess activating agent, fully dissolving, adding into the washed resin after dissolving, reacting for 1h, and washing with DMF.
4. And repeating the second step and the third step, and coupling the amino acids with the N-terminal Fmoc protection and the side chain protection in sequence from Lys to Tyr according to the main chain sequence.
5. After the synthesis is finished, the cracking is carried out, and the proportion of a cracking reagent is TFA: EDT (electro-thermal transfer coating): and Tis: TA: anisole: h2O = 80: 5: 1: 5: 5: 4 (volume ratio), 1g of peptide resin needs 10ml of cracking reagent, cracking for about 2h (120 r/min) at room temperature, precipitating with glacial methyl tert-butyl ether after cracking, and obtaining crude product as lower precipitate.
6. The crude peptide from the previous step was dissolved and purified on preparative HPLC with 0.1% TFA/acetonitrile.
7. And (4) freeze-drying after purification, taking out powder after freeze-drying, and subpackaging for quality inspection.
Example 7: synthesis of β -Ala-His-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg:
1. Fmoc-Arg (pbf) -CTC resin was obtained by coupling solid phase support 2-CTC resin with Fmoc-Arg (pbf) -OH in the presence of an activator system (HoBT, DIC).
2. The Fmoc protecting group on Fmoc-Arg (pbf) -CTC was removed with 20% piperidine and washed clean with DMF.
3. Weighing 3 times excess Fmoc-Arg (pbf) -OH and three times excess activating agent, adding a small amount of DMF to fully dissolve, adding the solution into a clean resin after the solution is completely dissolved, reacting for 1h, and washing with DMF.
4. And repeating the second step and the third step, and sequentially coupling amino acids with Fmoc protection at the N end and protected side chains from Arg to Ala in the main chain sequence.
5. After the synthesis is finished, the cracking is carried out, and the proportion of a cracking reagent is TFA: EDT (electro-thermal transfer coating): and Tis: TA: anisole: h2O = 80: 5: 1: 5: 5: 4 (volume ratio), 1g of peptide resin needs 10ml of cracking reagent, cracking for about 2h (120 r/min) at room temperature, precipitating with glacial methyl tert-butyl ether after cracking, and obtaining crude product as lower precipitate.
6. The crude peptide from the previous step was dissolved and purified on preparative HPLC with 0.1% TFA/acetonitrile.
7. And (4) freeze-drying after purification, taking out powder after freeze-drying, and performing subpackage quality inspection to obtain the target polypeptide with the purity of more than 95%.
The method of example 7 can also be used for the synthesis of the following polypeptides:
β-Ala-His-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg；
β-Ala-His-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg。
example 8: Pro-Ala-Lys- β -Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg:
1. Fmoc-Arg (pbf) -CTC resin was obtained by coupling solid phase support 2-CTC resin with Fmoc-Arg (pbf) -OH in the presence of an activator system (HoBT, DIC).
2. The Fmoc protecting group on Fmoc-Arg (pbf) -CTC was removed with 20% piperidine and washed clean with DMF.
3. Weighing 3 times excess Fmoc-Arg (pbf) -OH and three times excess activating agent, adding a small amount of DMF to fully dissolve, adding the solution into a clean resin after the solution is completely dissolved, reacting for 1h, and washing with DMF.
4. And repeating the second step and the third step, and coupling the amino acids with the N-terminal Fmoc protection and the side chain protection in sequence from Arg to Pro according to the main chain sequence.
5. After the synthesis is finished, the cracking is carried out, and the proportion of a cracking reagent is TFA: EDT (electro-thermal transfer coating): and Tis: TA: anisole: h2O = 80: 5: 1: 5: 5: 4 (volume ratio), 1g of peptide resin needs 10ml of cracking reagent, cracking for about 2h (120 r/min) at room temperature, precipitating with glacial methyl tert-butyl ether after cracking, and obtaining crude product as lower precipitate.
6. The crude peptide from the previous step was dissolved and purified on preparative HPLC with 0.1% TFA/acetonitrile.
7. And (4) freeze-drying after purification, taking out powder after freeze-drying, and performing subpackage quality inspection to obtain the target polypeptide with the purity of more than 95%.
Example 9: synthesis of Ala-Lys-Pro-beta-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg:
1. Fmoc-Arg (pbf) -CTC resin was obtained by coupling solid phase support 2-CTC resin with Fmoc-Arg (pbf) -OH in the presence of an activator system (HoBT, DIC).
2. The Fmoc protecting group on Fmoc-Arg (pbf) -CTC was removed with 20% piperidine and washed clean with DMF.
3. Weighing 3 times excess Fmoc-Arg (pbf) -OH and three times excess activating agent, adding a small amount of DMF to fully dissolve, adding the solution into a clean resin after the solution is completely dissolved, reacting for 1h, and washing with DMF.
4. And repeating the second step and the third step, and sequentially coupling amino acids with Fmoc protection at the N end and protected side chains from Arg to Ala in the main chain sequence.
5. After the synthesis is finished, the cracking is carried out, and the proportion of a cracking reagent is TFA: EDT (electro-thermal transfer coating): and Tis: TA: anisole: h2O = 80: 5: 1: 5: 5: 4 (volume ratio), 1g of peptide resin needs 10ml of cracking reagent, cracking for about 2h (120 r/min) at room temperature, precipitating with glacial methyl tert-butyl ether after cracking, and obtaining crude product as lower precipitate.
6. The crude peptide from the previous step was dissolved and purified on preparative HPLC with 0.1% TFA/acetonitrile.
7. And (4) freeze-drying after purification, taking out powder after freeze-drying, and performing subpackage quality inspection to obtain the target polypeptide with the purity of more than 95%.
Example 10: synthesis of Lys-Ala-Pro- β -Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg:
1. Fmoc-Arg (pbf) -CTC resin was obtained by coupling solid phase support 2-CTC resin with Fmoc-Arg (pbf) -OH in the presence of an activator system (HoBT, DIC).
2. The Fmoc protecting group on Fmoc-Arg (pbf) -CTC was removed with 20% piperidine and washed clean with DMF.
3. Weighing 3 times excess Fmoc-Arg (pbf) -OH and three times excess activating agent, adding a small amount of DMF to fully dissolve, adding the solution into a clean resin after the solution is completely dissolved, reacting for 1h, and washing with DMF.
4. And repeating the second step and the third step, and coupling the amino acids with the N-terminal Fmoc protection and the side chain protection in the sequence from Arg to Lys in the main chain sequence.
5. After the synthesis is finished, the cracking is carried out, and the proportion of a cracking reagent is TFA: EDT (electro-thermal transfer coating): and Tis: TA: anisole: h2O = 80: 5: 1: 5: 5: 4 (volume ratio), 1g of peptide resin needs 10ml of cracking reagent, cracking for about 2h (120 r/min) at room temperature, precipitating with glacial methyl tert-butyl ether after cracking, and obtaining crude product as lower precipitate.
6. The crude peptide from the previous step was dissolved and purified on preparative HPLC with 0.1% TFA/acetonitrile.
7. And (4) freeze-drying after purification, taking out powder after freeze-drying, and performing subpackage quality inspection to obtain the target polypeptide with the purity of more than 95%.
Example 11: synthesis of comparative polypeptides is described in patent application No.: 202110266153.0 (Lijiangxiong, Chinese patent, 2021).
Example 12: animal experiment method
The small animal monitor comprises: shenzhen, Shen honor Zhi science and technology Limited, VT 200.
SD rat: 180-200g of Chinese institute for food and drug identification.
Wire tying: meyer organism (M8507).
TTC staining solution: a source leaf organism (R24053).
Animal model operation steps:
experimental SD rats were weighed and anesthetized with chloral hydrate. After anesthesia, the four limbs of the rat are fixed and lie on the back, and the small animal monitor is connected to monitor important physiological indexes of the rat, such as body temperature, blood pressure, heart rate and the like. The rat neck was dehaired and a median incision of about 2cm length was made after sterilization with a 75% alcohol cotton ball, rat submandibular glands were dissected blunt, gland destruction was avoided as much as possible during dissection, then Common Carotid Artery (CCA) on the left side of the experimental rat was dissected blunt, and Internal Carotid Artery (ICA) and External Carotid Artery (ECA) were carefully dissected up along CCA. Damage to the vagus nerve was avoided during isolation. The CCA and the ICA are clamped and closed by two artery clamps respectively, the distal end of the ECA is cut off, a silica gel line bolt is inserted from an ECA 'port', the artery clamp is loosened briefly when the ICA is inserted to the ICA artery clamp, the ICA is clamped and closed again after the head end of the line bolt is inserted through the artery clamp, then the artery clamp on the ICA is loosened slightly and the line bolt is inserted continuously until the black mark of the line bolt is inserted through the bifurcation of the ECA and the ICA, and the ECA 'port' and the line bolt are tightly fastened by a suture after the head end of the line bolt is blocked on the middle cerebral artery so as to prevent the line bolt from 'withdrawing' or bleeding after the rat wakes up. The clamps on the CCA and ICA were loosened and removed to allow the tissue to recover in situ and an appropriate amount of penicillin was added dropwise to prevent wound infection. The suture is made with a medical suture needle with thread, sealed and sterilized again with iodophor. And observing the small animal monitor to judge whether the index of the experimental rat is normal, and then putting the rat on an electric blanket of the small animal to keep the body temperature until the rat is awakened and putting the rat in an animal cage.
The administration process comprises the following steps:
the drug to be tested is prepared into a solution of 3 mg/mL. The tail vein was dosed 1-2h after ligation.
When in administration, the rat is fixed by a fixer, a 1mL injector is used for absorbing the drug to be detected according to the dose of 3mg/kg, then the tail vein of the rat is injected, and the injection is slowly injected to reduce the load of the heart and lung of the experimental rat.
Animals were divided into different groups of six animals each.
Model group: after ligation, the tail vein is given with the same amount of normal saline;
the drug group S1 Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Leu-Ser-Ser-Ile-Glu-Ser-Asp-Val,
S2 Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro，
S4 β-Ala-His-Lys-Ala-Pro，
S5 Pro-Ala-Lys-β-Ala-His，
S6 Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-βAla-His，
s1-2 and 4-6, and the tail vein is administered after ligation;
the sham operation group: after isolating the Internal Carotid Artery (ICA) and the External Carotid Artery (ECA), ligation was not performed, and the tail vein was given an equal amount of physiological saline.
TTC staining and quantitative analysis:
after the behavioral observations, the experimental rats were euthanized and their brains were removed. The brain tissue is transversely cut into 6 slices with the thickness of 2mm, then the slices are transferred into TTC staining solution, incubated in an incubator at 37 ℃ for 10min in dark place, and photographed. The TTC stained brain tissue and the remaining small amount of brain tissue that was not subjected to TTC staining were then stored at-20 ℃.
Images after TTC staining were quantified using Image J software.
Cerebral infarction volume% = (total infarct area slice thickness)/(total brain slice area slice thickness) × 100%.
The experimental results are as follows:
1. the TTC quantification results are shown in table 1:
table 1: TTC quantitative results
| Numbering | 1 | 2 | 3 | 4 | 5 | 6 | average | STDEV |
| Model set | 0.41 | 0.40 | 0.39 | 0.42 | 0.38 | 0.34 | 0.39 | 0.03 |
| S1 | 0.19 | 0.22 | 0.15 | 0.15 | 0.11 | 0.13 | 0.16 | 0.04 |
| S2 | 0.34 | 0.25 | 0.22 | 0.12 | 0.14 | 0.10 | 0.20 | 0.09 |
| S4 | 0.23 | 0.16 | 0.14 | 0.17 | 0.12 | 0.12 | 0.16 | 0.04 |
| S5 | 0.12 | 0.14 | 0.11 | 0.18 | 0.09 | 0.23 | 0.15 | 0.05 |
| S6 | 0.16 | 0.13 | 0.11 | 0.19 | 0.14 | 0.18 | 0.15 | 0.03 |
| Artificial operation group | 0.05 | 0.04 | 0.04 | 0.05 | 0.04 | 0.04 | 0.04 | 0.01 |
S1： Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Leu-Ser-Ser-Ile-Glu-Ser-Asp-Val；
S2：Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro；
S4：β-Ala-His-Lys-Ala-Pro；
S5：Pro-Ala-Lys-β-Ala-His；
S6：Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-β-Ala-His。
As can be seen from the results in table 1, the cerebral ischemic rats of the model group had infarct volumes of 0.39 ± 0.03; cerebral ischemic rats injected with 3mg/kg Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-beta-Ala-His-Lys-Ala-Pro (S2) had infarct volume of 0.20 + -0.09, with no statistical difference compared to the results of 0.16 + -0.04 for Tyr-Gly-Arg-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Leu-Ser-Ser-Ile-Glu-Ser-Asp-Val (S1, NA1) and 0.16 + -0.04 for beta-Ala-His-Lys-Ala-Pro (S4). Compared with the Model group, S2 has obvious bioactivity and S2 is not statistically different from S1 and S4, wherein NA1 (Nerinetide) is a neuroprotective agent (Michael Tymianski and Jonathan D. Garman. Model systems and treatment reagents for treatment of neurological disease. 2015, US Patent, US8940699B2), can interfere with postsynaptic Density protein 95 (PSD-95), is realized by stopping the generation of intracellular NO free radicals, can reduce the infarct area of animal experimental (macaque) cerebral ischemia reperfusion in a preclinical ischemic stroke Model, and improves the functional prognosis, and is a good positive control product. Phll doctor studied the efficacy and safety of intravenous administration of the novel neuropeptide NA-1 (2.6 mg/kg) in patients with Acute Ischemic Stroke (AIS) following intravascular thrombectomy, and the results indicated that treatment with NA1 improved patient prognosis. (Michael D Hill et al, effectiveness and safety of a nitrile for the treatment of an acid electrochemical stress (ESCAPE-NA 1): a multicentre, double-blind, random controlled trial. Lance. Published connected February 20,2020). S2 is the combination of S4 and TAT polypeptide, and the activity of single molecule of S2 is improved from the molecular level.
As can also be seen from the results in table 1, the infarct volume in the saline-injected cerebral ischemic rats was 0.39 ± 0.03; cerebral ischemic rats injected with 3mg/kg Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys- β -Ala-His (S6) had infarct volume of 0.15. + -. 0.03, and were not statistically different from the results of 0.16. + -. 0.04 for Tyr-Gly-Arg-Lys-Arg-Arg-Gln-Arg-Arg-Lys-Leu-Ser-Ser-Ile-Glu-Ser-Asp-Val (S1, NA1) and 0.15. + -. 0.05 for Pro-Ala-Lys- β Ala-His (S5). Compared with the sham group, S6 had significant bioactivity and S6 was not statistically different from S1 and S5. S6 is the binding of S5 to TAT polypeptide, which also indicates that, on a molecular level, the activity of S6 as a single molecule is increased.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. A polypeptide combination comprising a polypeptide fragment A and a polypeptide fragment B, wherein the polypeptide fragment A is Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg, and the polypeptide fragment B is Pro-Ala-Lys, Ala-Lys-Pro, Lys-Ala-Pro, Pro-Ala, Ala-Lys, Lys-Pro, Lys-Ala, Ala-Pro, Ala-Arg-Pro-Ala-Lys, Ala-Arg-Ala-Lys-Pro, Arg-Lys-Ala-Pro, Arg-Lys-Pro, Arg-Ala-Pro, Gly-Arg-Pro-Ala-Lys, or Arg-Ala-Pro, One or more of Gly-Arg-Ala-Lys-Pro, Gly-Arg-Lys-Ala-Pro, Gln-Arg-Pro-Ala-Lys, Gln-Arg-Ala-Lys-Pro and Gln-Arg-Lys-Ala-Pro, and the C terminal and/or N terminal of the composition has carnosine, anserine or sercarnosine.
2. The combination polypeptide of claim 1, wherein the polypeptide fragment B is composed of one or more of Pro-Ala-Lys, Ala-Lys-Pro, Lys-Ala-Pro, Pro-Ala, Ala-Lys, Lys-Pro, Lys-Ala, and Ala-Pro and has carnosine, anserine, or serosine at its C-terminus and/or N-terminus, comprising:
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Pro-Ala，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Pro-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Lys-Ala，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Pro-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Pro-Ala，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Pro-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Lys-Ala，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Pro-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Pro-Ala，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Pro-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Lys-Ala，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Pro-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Pro-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Pro-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Lys-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Pro-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Pro-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Pro-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Lys-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Pro-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Pro-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Pro-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Lys-Ala-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Pro-Lys-Ala-Pro-β-Ala-His，
β-Ala-His-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Pro-Ala-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Pro-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Lys-Ala-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Pro-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Pro-Ala-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Pro-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Lys-Ala-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Pro-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Pro-Ala-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Pro-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Lys-Ala-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Pro-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Pro-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Pro-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Lys-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Pro-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Pro-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Pro-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Lys-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Pro-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Pro-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Ala-Pro-β-Ala-Hi-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Args，
Lys-Ala-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Pro-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Lys-Ala-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Pro-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
wherein His is His, 1-Methyl-His or 3-Methyl-His.
3. The combination polypeptide of claim 1, wherein the polypeptide fragment B is a hexapeptide consisting of one or two of Pro-Ala-Lys, Ala-Lys-Pro, and Lys-Ala-Pro and has carnosine, anserine, or sercarnosine at the C-terminus and/or N-terminus, comprising:
Pro-Ala-Lys-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His -Ala-Lys-Pro-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His -Pro-Ala-Lys-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His -Lys-Ala-Pro-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His -Lys-Ala-Pro-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His -Lys-Ala-Pro-Lys-Ala-Pro，
β-Ala-His-Pro-Ala-Lys-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-Lys-Ala-Pro-β-Ala-His，
wherein His is His, 1-Methyl-His or 3-Methyl-His.
4. The combination polypeptide of claim 1, comprising:
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro-β-Ala-His，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro-β-Ala-His，
β-Ala-His-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
β-Ala-His-Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
wherein His is His, 1-Methyl-His or 3-Methyl-His.
5. The combination polypeptide of claim 1, comprising:
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Pro-Ala-Lys，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-β-Ala-His，
β-Ala-His-Pro-Ala-Lys-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Pro-Ala-Lys-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
wherein His is His, 1-Methyl-His or 3-Methyl-His.
6. The combination polypeptide of claim 1, comprising:
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Ala-Lys-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Ala-Lys-Pro-β-Ala-His，
β-Ala-His-Ala-Lys-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Ala-Lys-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
wherein His is His, 1-Methyl-His or 3-Methyl-His.
7. The combination polypeptide of claim 1, comprising:
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Lys-Ala-Pro-β-Ala-His，
β-Ala-His-Lys-Ala-Pro-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
Lys-Ala-Pro-β-Ala-His-Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg，
wherein His is His, 1-Methyl-His or 3-Methyl-His.
8. The combination polypeptide of claim 1, comprising:
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-β-Ala-His-Lys-Ala-Pro，
Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-Pro-Ala-Lys-β-Ala-His。
9. use of a combination polypeptide as claimed in any one of claims 1 to 8 and salts thereof in the manufacture of a medicament.
10. The use according to claim 9, characterized in that the combined polypeptide and its salts are used for the preparation of medicaments for the treatment of ischemic stroke, hemorrhagic stroke, brain trauma, alzheimer's disease, parkinson's disease or other neurodegenerative diseases.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110873583.9A CN113735939A (en) | 2021-07-30 | 2021-07-30 | Combined polypeptide and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110873583.9A CN113735939A (en) | 2021-07-30 | 2021-07-30 | Combined polypeptide and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113735939A true CN113735939A (en) | 2021-12-03 |
Family
ID=78729558
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110873583.9A Pending CN113735939A (en) | 2021-07-30 | 2021-07-30 | Combined polypeptide and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113735939A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113150065A (en) * | 2021-03-11 | 2021-07-23 | 武汉英纳氏药业有限公司 | Synthetic peptide and application thereof |
| CN115677668A (en) * | 2022-10-28 | 2023-02-03 | 东南大学 | Anserine derivatives and their preparation methods and applications |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101678068A (en) * | 2007-03-02 | 2010-03-24 | 阿尔伯维塔公司 | Do not suppress N type calcium channel and treat apoplexy and other diseases |
| CN103936838A (en) * | 2014-04-10 | 2014-07-23 | 武汉启瑞科技发展有限公司 | Micro-molecule polypeptide TAT-p53DM and application thereof to preparing medicine for treating or preventing ischemic stroke |
| CN106916210A (en) * | 2017-03-13 | 2017-07-04 | 河北科技大学 | A kind of polypeptide and its application with treating cerebral ischemia |
| CN106928322A (en) * | 2017-03-13 | 2017-07-07 | 河北科技大学 | A kind of fused polypeptide and its application with treating cerebral ischemia |
| CN109293743A (en) * | 2018-10-30 | 2019-02-01 | 河北科技大学 | A novel fusion polypeptide with anti-cerebral ischemia effect and its application |
| CN113121641A (en) * | 2021-03-11 | 2021-07-16 | 武汉英纳氏药业有限公司 | Multifunctional polypeptide and application thereof in medicine field |
| CN113150065A (en) * | 2021-03-11 | 2021-07-23 | 武汉英纳氏药业有限公司 | Synthetic peptide and application thereof |
-
2021
- 2021-07-30 CN CN202110873583.9A patent/CN113735939A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101678068A (en) * | 2007-03-02 | 2010-03-24 | 阿尔伯维塔公司 | Do not suppress N type calcium channel and treat apoplexy and other diseases |
| CN103936838A (en) * | 2014-04-10 | 2014-07-23 | 武汉启瑞科技发展有限公司 | Micro-molecule polypeptide TAT-p53DM and application thereof to preparing medicine for treating or preventing ischemic stroke |
| CN106916210A (en) * | 2017-03-13 | 2017-07-04 | 河北科技大学 | A kind of polypeptide and its application with treating cerebral ischemia |
| CN106928322A (en) * | 2017-03-13 | 2017-07-07 | 河北科技大学 | A kind of fused polypeptide and its application with treating cerebral ischemia |
| CN109293743A (en) * | 2018-10-30 | 2019-02-01 | 河北科技大学 | A novel fusion polypeptide with anti-cerebral ischemia effect and its application |
| CN113121641A (en) * | 2021-03-11 | 2021-07-16 | 武汉英纳氏药业有限公司 | Multifunctional polypeptide and application thereof in medicine field |
| CN113150065A (en) * | 2021-03-11 | 2021-07-23 | 武汉英纳氏药业有限公司 | Synthetic peptide and application thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113150065A (en) * | 2021-03-11 | 2021-07-23 | 武汉英纳氏药业有限公司 | Synthetic peptide and application thereof |
| CN113150065B (en) * | 2021-03-11 | 2022-07-08 | 武汉英纳氏药业有限公司 | Synthetic peptide and application thereof |
| CN115677668A (en) * | 2022-10-28 | 2023-02-03 | 东南大学 | Anserine derivatives and their preparation methods and applications |
| CN115677668B (en) * | 2022-10-28 | 2023-12-05 | 东南大学 | Anserine derivative, and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113121641B (en) | A class of multifunctional polypeptides and their applications in the field of medicine | |
| JP2793719B2 (en) | Cytokine inhibitor | |
| CN113150065B (en) | Synthetic peptide and application thereof | |
| USRE43309E1 (en) | Immunoregulatory compositions | |
| TWI882141B (en) | Peptide for repairing mucosal damage or skin trauma and its application | |
| CN113735939A (en) | Combined polypeptide and application thereof | |
| CN101497651A (en) | Compound with thrombolytic activity, preparation method and application thereof | |
| CN113735938B (en) | A neuroprotective polypeptide compound and its application | |
| JP4933440B2 (en) | Pharmaceutical composition for transdermal delivery | |
| JPH05503103A (en) | Anti-inflammatory peptides that suppress vascular leakage in damaged tissues and methods for treating the tissues | |
| EP3162811B1 (en) | Peptide as absorption enhancer and composition containing same | |
| RU2537560C2 (en) | Tetrapeptide and preparation, possessing cerebroprotective and antiamnestic activities (versions) | |
| JP2024545747A (en) | Pentapeptides and their uses | |
| CN114106100A (en) | Polypeptide for repairing skin wound or mucosal injury and application thereof | |
| JP3002725B2 (en) | Pharmaceutical composition comprising fibroblast growth factor-related peptide | |
| CN120265642A (en) | Compound, preparation method and application thereof | |
| CN111214649A (en) | Application of BM23 peptide in preparing medicine for treating ischemic cerebrovascular disease | |
| CN119176850A (en) | Neuroprotection agent capable of being combined with rt-PA and application thereof | |
| EP0147194B1 (en) | Compositions for treating cerebral ischemia | |
| CN118512430A (en) | Application of midodrine hydrochloride in medicine for alleviating intestinal ischemia-reperfusion injury | |
| JP2002179699A (en) | Novel peptide and composition using the peptide |
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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20211203 |