CN101273058A - 增强肽酶抗性的glp-1(胰高血糖素样肽-1)融合多肽 - Google Patents
增强肽酶抗性的glp-1(胰高血糖素样肽-1)融合多肽 Download PDFInfo
- Publication number
- CN101273058A CN101273058A CNA2006800350367A CN200680035036A CN101273058A CN 101273058 A CN101273058 A CN 101273058A CN A2006800350367 A CNA2006800350367 A CN A2006800350367A CN 200680035036 A CN200680035036 A CN 200680035036A CN 101273058 A CN101273058 A CN 101273058A
- Authority
- CN
- China
- Prior art keywords
- sequence
- peptide
- ala
- seq
- glu
- 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 369
- DTHNMHAUYICORS-KTKZVXAJSA-N Glucagon-like peptide 1 Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1N=CNC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=CC=C1 DTHNMHAUYICORS-KTKZVXAJSA-N 0.000 title claims abstract description 120
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 37
- 102400000322 Glucagon-like peptide 1 Human genes 0.000 title claims abstract 9
- 230000004927 fusion Effects 0.000 title abstract description 8
- 101800000224 Glucagon-like peptide 1 Proteins 0.000 title description 114
- 229920001184 polypeptide Polymers 0.000 title description 18
- 102000035195 Peptidases Human genes 0.000 title description 9
- 108091005804 Peptidases Proteins 0.000 title description 9
- 230000001965 increasing effect Effects 0.000 title description 6
- 235000019833 protease Nutrition 0.000 title description 4
- 150000001413 amino acids Chemical class 0.000 claims abstract description 85
- 239000012634 fragment Substances 0.000 claims abstract description 47
- TWSALRJGPBVBQU-PKQQPRCHSA-N glucagon-like peptide 2 Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(O)=O)[C@@H](C)CC)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCSC)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CO)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)[C@@H](C)CC)C1=CC=CC=C1 TWSALRJGPBVBQU-PKQQPRCHSA-N 0.000 claims abstract description 33
- 101800000221 Glucagon-like peptide 2 Proteins 0.000 claims abstract description 29
- 239000003814 drug Substances 0.000 claims abstract description 23
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 22
- 201000010099 disease Diseases 0.000 claims abstract description 16
- 208000001072 type 2 diabetes mellitus Diseases 0.000 claims abstract description 10
- 238000002360 preparation method Methods 0.000 claims abstract description 7
- 101710198884 GATA-type zinc finger protein 1 Proteins 0.000 claims abstract 7
- 102400000326 Glucagon-like peptide 2 Human genes 0.000 claims abstract 2
- 229940024606 amino acid Drugs 0.000 claims description 84
- 235000001014 amino acid Nutrition 0.000 claims description 82
- 210000004027 cell Anatomy 0.000 claims description 77
- 230000000799 fusogenic effect Effects 0.000 claims description 64
- GCYXWQUSHADNBF-AAEALURTSA-N preproglucagon 78-108 Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1N=CNC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=CC=C1 GCYXWQUSHADNBF-AAEALURTSA-N 0.000 claims description 58
- -1 deaminize-Histidine Chemical compound 0.000 claims description 56
- 101800004266 Glucagon-like peptide 1(7-37) Proteins 0.000 claims description 54
- 238000011282 treatment Methods 0.000 claims description 48
- 210000004369 blood Anatomy 0.000 claims description 37
- 239000008280 blood Substances 0.000 claims description 37
- 229960002885 histidine Drugs 0.000 claims description 37
- 238000000034 method Methods 0.000 claims description 33
- 108090000623 proteins and genes Proteins 0.000 claims description 31
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 claims description 29
- 125000003295 alanine group Chemical group N[C@@H](C)C(=O)* 0.000 claims description 29
- 150000001732 carboxylic acid derivatives Chemical class 0.000 claims description 27
- 230000004048 modification Effects 0.000 claims description 24
- 238000012986 modification Methods 0.000 claims description 24
- 235000018102 proteins Nutrition 0.000 claims description 21
- 102000004169 proteins and genes Human genes 0.000 claims description 21
- 125000000539 amino acid group Chemical group 0.000 claims description 20
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 19
- 239000002773 nucleotide Substances 0.000 claims description 18
- 125000003729 nucleotide group Chemical group 0.000 claims description 18
- 241001465754 Metazoa Species 0.000 claims description 15
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 15
- 102000004877 Insulin Human genes 0.000 claims description 14
- 108090001061 Insulin Proteins 0.000 claims description 14
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 claims description 14
- 230000008034 disappearance Effects 0.000 claims description 13
- 238000006243 chemical reaction Methods 0.000 claims description 12
- 230000037396 body weight Effects 0.000 claims description 10
- UYEGXSNFZXWSDV-BYPYZUCNSA-N (2s)-3-(2-amino-1h-imidazol-5-yl)-2-azaniumylpropanoate Chemical compound OC(=O)[C@@H](N)CC1=CNC(N)=N1 UYEGXSNFZXWSDV-BYPYZUCNSA-N 0.000 claims description 7
- WOXWUZCRWJWTRT-UHFFFAOYSA-N 1-amino-1-cyclohexanecarboxylic acid Chemical compound OC(=O)C1(N)CCCCC1 WOXWUZCRWJWTRT-UHFFFAOYSA-N 0.000 claims description 7
- NILQLFBWTXNUOE-UHFFFAOYSA-N 1-aminocyclopentanecarboxylic acid Chemical compound OC(=O)C1(N)CCCC1 NILQLFBWTXNUOE-UHFFFAOYSA-N 0.000 claims description 7
- HNDVDQJCIGZPNO-RXMQYKEDSA-N D-histidine Chemical compound OC(=O)[C@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-RXMQYKEDSA-N 0.000 claims description 7
- 229930195721 D-histidine Natural products 0.000 claims description 7
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 claims description 7
- KBOJOGQFRVVWBH-ZETCQYMHSA-N N-acetyl-L-histidine Chemical compound CC(=O)N[C@H](C(O)=O)CC1=CN=CN1 KBOJOGQFRVVWBH-ZETCQYMHSA-N 0.000 claims description 7
- 229940125396 insulin Drugs 0.000 claims description 7
- 125000001360 methionine group Chemical group N[C@@H](CCSC)C(=O)* 0.000 claims description 6
- 210000000496 pancreas Anatomy 0.000 claims description 6
- 125000001500 prolyl group Chemical group [H]N1C([H])(C(=O)[*])C([H])([H])C([H])([H])C1([H])[H] 0.000 claims description 5
- MPVGZUDRATZEEE-YFKPBYRVSA-N (2s)-2-amino-3-(3-hydroxyimidazol-4-yl)propanoic acid Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1O MPVGZUDRATZEEE-YFKPBYRVSA-N 0.000 claims description 3
- 206010015995 Eyelid ptosis Diseases 0.000 claims description 3
- 201000003004 ptosis Diseases 0.000 claims description 3
- 238000010532 solid phase synthesis reaction Methods 0.000 claims description 3
- 239000004215 Carbon black (E152) Substances 0.000 claims description 2
- 229930195733 hydrocarbon Natural products 0.000 claims description 2
- 125000001165 hydrophobic group Chemical group 0.000 claims description 2
- 230000000926 neurological effect Effects 0.000 claims description 2
- 125000000487 histidyl group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 claims 2
- 102000014914 Carrier Proteins Human genes 0.000 claims 1
- 108010078791 Carrier Proteins Proteins 0.000 claims 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims 1
- 125000000729 N-terminal amino-acid group Chemical group 0.000 claims 1
- 239000005864 Sulphur Substances 0.000 claims 1
- 102000002070 Transferrins Human genes 0.000 claims 1
- 108010015865 Transferrins Proteins 0.000 claims 1
- 210000004899 c-terminal region Anatomy 0.000 claims 1
- 238000000855 fermentation Methods 0.000 claims 1
- 230000004151 fermentation Effects 0.000 claims 1
- 150000002505 iron Chemical class 0.000 claims 1
- 230000000813 microbial effect Effects 0.000 claims 1
- 238000010647 peptide synthesis reaction Methods 0.000 claims 1
- 230000001568 sexual effect Effects 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 37
- 230000036961 partial effect Effects 0.000 abstract description 7
- 208000035475 disorder Diseases 0.000 abstract description 5
- 206010067584 Type 1 diabetes mellitus Diseases 0.000 abstract description 3
- 238000001727 in vivo Methods 0.000 abstract description 3
- 208000015122 neurodegenerative disease Diseases 0.000 abstract description 3
- 102000016622 Dipeptidyl Peptidase 4 Human genes 0.000 abstract 1
- 101000930822 Giardia intestinalis Dipeptidyl-peptidase 4 Proteins 0.000 abstract 1
- 208000034615 apoptosis-related disease Diseases 0.000 abstract 1
- 102100040918 Pro-glucagon Human genes 0.000 description 121
- 241000699666 Mus <mouse, genus> Species 0.000 description 48
- 108010080629 tryptophan-leucine Proteins 0.000 description 45
- KSCVLGXNQXKUAR-JYJNAYRXSA-N Tyr-Leu-Glu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O KSCVLGXNQXKUAR-JYJNAYRXSA-N 0.000 description 43
- LLWQVJNHMYBLLK-CDMKHQONSA-N Gly-Thr-Phe Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O LLWQVJNHMYBLLK-CDMKHQONSA-N 0.000 description 41
- IVDFVBVIVLJJHR-LKXGYXEUSA-N Thr-Ser-Asp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O IVDFVBVIVLJJHR-LKXGYXEUSA-N 0.000 description 41
- PZTZYZUTCPZWJH-FXQIFTODSA-N Val-Ser-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)O)N PZTZYZUTCPZWJH-FXQIFTODSA-N 0.000 description 41
- WZPIKDWQVRTATP-SYWGBEHUSA-N Ile-Ala-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](C)NC(=O)[C@@H](N)[C@@H](C)CC)C(O)=O)=CNC2=C1 WZPIKDWQVRTATP-SYWGBEHUSA-N 0.000 description 40
- YQFZRHYZLARWDY-IHRRRGAJSA-N Leu-Val-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN YQFZRHYZLARWDY-IHRRRGAJSA-N 0.000 description 40
- DUTMKEAPLLUGNO-JYJNAYRXSA-N Lys-Glu-Phe Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O DUTMKEAPLLUGNO-JYJNAYRXSA-N 0.000 description 40
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 39
- RJIVPOXLQFJRTG-LURJTMIESA-N Gly-Arg-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CCCN=C(N)N RJIVPOXLQFJRTG-LURJTMIESA-N 0.000 description 38
- 150000003839 salts Chemical class 0.000 description 37
- YJIUYQKQBBQYHZ-ACZMJKKPSA-N Gln-Ala-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O YJIUYQKQBBQYHZ-ACZMJKKPSA-N 0.000 description 36
- AFPFGFUGETYOSY-HGNGGELXSA-N His-Ala-Glu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O AFPFGFUGETYOSY-HGNGGELXSA-N 0.000 description 36
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 36
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 35
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 34
- 239000008103 glucose Substances 0.000 description 34
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 33
- 239000002253 acid Substances 0.000 description 30
- 239000000203 mixture Substances 0.000 description 27
- ODKSFYDXXFIFQN-BYPYZUCNSA-N L-arginine Chemical compound OC(=O)[C@@H](N)CCCN=C(N)N ODKSFYDXXFIFQN-BYPYZUCNSA-N 0.000 description 26
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 24
- LTFLDDDGWOVIHY-NAKRPEOUSA-N Val-Ala-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](C)NC(=O)[C@H](C(C)C)N LTFLDDDGWOVIHY-NAKRPEOUSA-N 0.000 description 22
- TTXYKSADPSNOIF-IHRRRGAJSA-N Arg-Asp-Phe Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O TTXYKSADPSNOIF-IHRRRGAJSA-N 0.000 description 21
- 102100025012 Dipeptidyl peptidase 4 Human genes 0.000 description 21
- 239000011159 matrix material Substances 0.000 description 21
- FRKBNXCFJBPJOL-GUBZILKMSA-N Pro-Glu-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O FRKBNXCFJBPJOL-GUBZILKMSA-N 0.000 description 20
- 238000002347 injection Methods 0.000 description 20
- 239000007924 injection Substances 0.000 description 20
- 229940090044 injection Drugs 0.000 description 20
- 150000001412 amines Chemical class 0.000 description 19
- 210000002381 plasma Anatomy 0.000 description 19
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerol Natural products OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 17
- FUOOLUPWFVMBKG-UHFFFAOYSA-N 2-Aminoisobutyric acid Chemical compound CC(C)(N)C(O)=O FUOOLUPWFVMBKG-UHFFFAOYSA-N 0.000 description 16
- 108010067722 Dipeptidyl Peptidase 4 Proteins 0.000 description 16
- 150000001875 compounds Chemical class 0.000 description 16
- 125000003275 alpha amino acid group Chemical group 0.000 description 15
- 108010068380 arginylarginine Proteins 0.000 description 15
- 206010012601 diabetes mellitus Diseases 0.000 description 15
- 238000009472 formulation Methods 0.000 description 15
- 108020004414 DNA Proteins 0.000 description 14
- 238000012360 testing method Methods 0.000 description 14
- 239000000052 vinegar Substances 0.000 description 14
- 235000021419 vinegar Nutrition 0.000 description 14
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 13
- WKOBSJOZRJJVRZ-FXQIFTODSA-N Ala-Glu-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O WKOBSJOZRJJVRZ-FXQIFTODSA-N 0.000 description 13
- 239000004475 Arginine Substances 0.000 description 13
- KIEICAOUSNYOLM-NRPADANISA-N Glu-Val-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O KIEICAOUSNYOLM-NRPADANISA-N 0.000 description 13
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 13
- SZTTYWIUCGSURQ-AUTRQRHGSA-N Val-Glu-Glu Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O SZTTYWIUCGSURQ-AUTRQRHGSA-N 0.000 description 13
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 13
- 230000000875 corresponding effect Effects 0.000 description 13
- XUYPXLNMDZIRQH-LURJTMIESA-N N-acetyl-L-methionine Chemical compound CSCC[C@@H](C(O)=O)NC(C)=O XUYPXLNMDZIRQH-LURJTMIESA-N 0.000 description 12
- QARPMYDMYVLFMW-KKUMJFAQSA-N Phe-Pro-Glu Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(O)=O)C1=CC=CC=C1 QARPMYDMYVLFMW-KKUMJFAQSA-N 0.000 description 12
- 239000003153 chemical reaction reagent Substances 0.000 description 12
- 239000000725 suspension Substances 0.000 description 12
- 238000001890 transfection Methods 0.000 description 12
- BABSVXFGKFLIGW-UWVGGRQHSA-N Leu-Gly-Arg Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCNC(N)=N BABSVXFGKFLIGW-UWVGGRQHSA-N 0.000 description 11
- 108091028043 Nucleic acid sequence Proteins 0.000 description 11
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 11
- 230000004071 biological effect Effects 0.000 description 11
- 238000002474 experimental method Methods 0.000 description 11
- 108010050848 glycylleucine Proteins 0.000 description 11
- 239000007788 liquid Substances 0.000 description 11
- 229930182817 methionine Natural products 0.000 description 11
- 239000000047 product Substances 0.000 description 11
- 230000004044 response Effects 0.000 description 11
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 10
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 10
- OYJCVIGKMXUVKB-GARJFASQSA-N Ala-Leu-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N OYJCVIGKMXUVKB-GARJFASQSA-N 0.000 description 9
- 108010011459 Exenatide Proteins 0.000 description 9
- 101800004295 Glucagon-like peptide 1(7-36) Proteins 0.000 description 9
- JUFFVKRROAPVBI-PVOYSMBESA-N chembl1210015 Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(=O)N[C@H]1[C@@H]([C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO[C@]3(O[C@@H](C[C@H](O)[C@H](O)CO)[C@H](NC(C)=O)[C@@H](O)C3)C(O)=O)O2)O)[C@@H](CO)O1)NC(C)=O)C(=O)NCC(=O)NCC(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CO)C(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCSC)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)CNC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=CC=C1 JUFFVKRROAPVBI-PVOYSMBESA-N 0.000 description 9
- 229960001519 exenatide Drugs 0.000 description 9
- 210000002966 serum Anatomy 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- NGJOFQZEYQGZMB-KTKZVXAJSA-N (4S)-5-[[2-[[(2S,3R)-1-[[(2S)-1-[[(2S,3R)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[2-[[(2S)-5-amino-1-[[(2S)-1-[[(2S)-1-[[(2S)-6-amino-1-[[(2S)-1-[[(2S)-1-[[(2S,3S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-6-amino-1-[[2-[[(1S)-4-carbamimidamido-1-carboxybutyl]amino]-2-oxoethyl]amino]-1-oxohexan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-3-(1H-indol-3-yl)-1-oxopropan-2-yl]amino]-1-oxopropan-2-yl]amino]-3-methyl-1-oxopentan-2-yl]amino]-1-oxo-3-phenylpropan-2-yl]amino]-4-carboxy-1-oxobutan-2-yl]amino]-1-oxohexan-2-yl]amino]-1-oxopropan-2-yl]amino]-1-oxopropan-2-yl]amino]-1,5-dioxopentan-2-yl]amino]-2-oxoethyl]amino]-4-carboxy-1-oxobutan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-3-(4-hydroxyphenyl)-1-oxopropan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-3-carboxy-1-oxopropan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-3-hydroxy-1-oxobutan-2-yl]amino]-1-oxo-3-phenylpropan-2-yl]amino]-3-hydroxy-1-oxobutan-2-yl]amino]-2-oxoethyl]amino]-4-[[(2S)-2-[[(2S)-2-amino-3-(1H-imidazol-4-yl)propanoyl]amino]propanoyl]amino]-5-oxopentanoic acid Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=CC=C1 NGJOFQZEYQGZMB-KTKZVXAJSA-N 0.000 description 8
- DKJPOZOEBONHFS-ZLUOBGJFSA-N Ala-Ala-Asp Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O DKJPOZOEBONHFS-ZLUOBGJFSA-N 0.000 description 8
- BIOCIVSVEDFKDJ-GUBZILKMSA-N Arg-Arg-Asp Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O BIOCIVSVEDFKDJ-GUBZILKMSA-N 0.000 description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 8
- 108060003199 Glucagon Proteins 0.000 description 8
- 108010054147 Hemoglobins Proteins 0.000 description 8
- 102000001554 Hemoglobins Human genes 0.000 description 8
- 108091016366 Histone-lysine N-methyltransferase EHMT1 Proteins 0.000 description 8
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 8
- 108010079364 N-glycylalanine Proteins 0.000 description 8
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 8
- 108010047857 aspartylglycine Proteins 0.000 description 8
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 8
- 239000003795 chemical substances by application Substances 0.000 description 8
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 8
- SLNCSSWAIDUUGF-LSJOCFKGSA-N Arg-His-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C)C(O)=O SLNCSSWAIDUUGF-LSJOCFKGSA-N 0.000 description 7
- 102000051325 Glucagon Human genes 0.000 description 7
- 239000004471 Glycine Substances 0.000 description 7
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 7
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 7
- 108010076504 Protein Sorting Signals Proteins 0.000 description 7
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 7
- 239000004473 Threonine Substances 0.000 description 7
- 150000001720 carbohydrates Chemical class 0.000 description 7
- 235000014633 carbohydrates Nutrition 0.000 description 7
- WDRMVIMVHHWVBI-STCSGHEYSA-N chembl1222074 Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)N[C@@H](CC=1NC=NC=1)C(O)=O)[C@@H](C)O)[C@H](C)O)C(C)C)C1=CC=CC=C1 WDRMVIMVHHWVBI-STCSGHEYSA-N 0.000 description 7
- 230000014509 gene expression Effects 0.000 description 7
- MASNOZXLGMXCHN-ZLPAWPGGSA-N glucagon Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O)C(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 MASNOZXLGMXCHN-ZLPAWPGGSA-N 0.000 description 7
- 229960004666 glucagon Drugs 0.000 description 7
- 235000011187 glycerol Nutrition 0.000 description 7
- 229960001153 serine Drugs 0.000 description 7
- 238000010254 subcutaneous injection Methods 0.000 description 7
- 239000007929 subcutaneous injection Substances 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- 229960002898 threonine Drugs 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- ATVYZJGOZLVXDK-IUCAKERBSA-N Glu-Leu-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O ATVYZJGOZLVXDK-IUCAKERBSA-N 0.000 description 6
- JSLVAHYTAJJEQH-QWRGUYRKSA-N Gly-Ser-Phe Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 JSLVAHYTAJJEQH-QWRGUYRKSA-N 0.000 description 6
- 102100035043 Histone-lysine N-methyltransferase EHMT1 Human genes 0.000 description 6
- JRHFQUPIZOYKQP-KBIXCLLPSA-N Ile-Ala-Glu Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCC(O)=O JRHFQUPIZOYKQP-KBIXCLLPSA-N 0.000 description 6
- OUUCIIJSBIBCHB-ZPFDUUQYSA-N Ile-Leu-Asp Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O OUUCIIJSBIBCHB-ZPFDUUQYSA-N 0.000 description 6
- YWCJXQKATPNPOE-UKJIMTQDSA-N Ile-Val-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N YWCJXQKATPNPOE-UKJIMTQDSA-N 0.000 description 6
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 6
- 125000000174 L-prolyl group Chemical group [H]N1C([H])([H])C([H])([H])C([H])([H])[C@@]1([H])C(*)=O 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- FTVRVZNYIYWJGB-ACZMJKKPSA-N Ser-Asp-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O FTVRVZNYIYWJGB-ACZMJKKPSA-N 0.000 description 6
- 239000013543 active substance Substances 0.000 description 6
- 125000003368 amide group Chemical group 0.000 description 6
- 239000002585 base Substances 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 6
- 238000006731 degradation reaction Methods 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 238000006471 dimerization reaction Methods 0.000 description 6
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 230000017854 proteolysis Effects 0.000 description 6
- 230000009467 reduction Effects 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- FGWRYRAVBVOHIB-XIRDDKMYSA-N Gln-Pro-Trp Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CCC(=O)N)N)C(=O)N[C@@H](CC2=CNC3=CC=CC=C32)C(=O)O FGWRYRAVBVOHIB-XIRDDKMYSA-N 0.000 description 5
- CQZDZKRHFWJXDF-WDSKDSINSA-N Gly-Gln-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CN CQZDZKRHFWJXDF-WDSKDSINSA-N 0.000 description 5
- IBMVEYRWAWIOTN-UHFFFAOYSA-N L-Leucyl-L-Arginyl-L-Proline Natural products CC(C)CC(N)C(=O)NC(CCCN=C(N)N)C(=O)N1CCCC1C(O)=O IBMVEYRWAWIOTN-UHFFFAOYSA-N 0.000 description 5
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 5
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 5
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 5
- 239000000654 additive Substances 0.000 description 5
- 235000004279 alanine Nutrition 0.000 description 5
- 108010087924 alanylproline Proteins 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 108010029539 arginyl-prolyl-proline Proteins 0.000 description 5
- 230000027455 binding Effects 0.000 description 5
- 230000000975 bioactive effect Effects 0.000 description 5
- 230000015556 catabolic process Effects 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 235000012631 food intake Nutrition 0.000 description 5
- 230000006870 function Effects 0.000 description 5
- 230000036571 hydration Effects 0.000 description 5
- 238000006703 hydration reaction Methods 0.000 description 5
- MGXWVYUBJRZYPE-YUGYIWNOSA-N incretin Chemical class C([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1C=CC(O)=CC=1)[C@@H](C)O)[C@@H](C)CC)C1=CC=C(O)C=C1 MGXWVYUBJRZYPE-YUGYIWNOSA-N 0.000 description 5
- 239000000859 incretin Substances 0.000 description 5
- 229910052500 inorganic mineral Inorganic materials 0.000 description 5
- 229910052742 iron Inorganic materials 0.000 description 5
- 108010073472 leucyl-prolyl-proline Proteins 0.000 description 5
- 239000011707 mineral Substances 0.000 description 5
- 235000010755 mineral Nutrition 0.000 description 5
- 235000020925 non fasting Nutrition 0.000 description 5
- 108020004707 nucleic acids Proteins 0.000 description 5
- 150000007523 nucleic acids Chemical class 0.000 description 5
- 102000039446 nucleic acids Human genes 0.000 description 5
- 229920001451 polypropylene glycol Polymers 0.000 description 5
- 230000028327 secretion Effects 0.000 description 5
- 150000005846 sugar alcohols Chemical class 0.000 description 5
- 238000012546 transfer Methods 0.000 description 5
- 108010029384 tryptophyl-histidine Proteins 0.000 description 5
- HSKCCNHLWZSIDN-LZVKGNEYSA-N (2s)-2-[[(2s)-1-[(2s)-2-[[2-[[(2r)-2-[[(2s)-5-(diaminomethylideneamino)-2-[[(2s)-pyrrolidine-2-carbonyl]amino]pentanoyl]amino]-3-sulfanylpropanoyl]amino]acetyl]amino]-3-methylbutanoyl]pyrrolidine-2-carbonyl]amino]butanedioic acid Chemical compound N([C@@H](CCCN=C(N)N)C(=O)N[C@@H](CS)C(=O)NCC(=O)N[C@@H](C(C)C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CC(O)=O)C(O)=O)C(=O)[C@@H]1CCCN1 HSKCCNHLWZSIDN-LZVKGNEYSA-N 0.000 description 4
- FJKROLUGYXJWQN-UHFFFAOYSA-N 4-hydroxybenzoic acid Chemical compound OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 4
- DVWVZSJAYIJZFI-FXQIFTODSA-N Ala-Arg-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(O)=O DVWVZSJAYIJZFI-FXQIFTODSA-N 0.000 description 4
- MKZCBYZBCINNJN-DLOVCJGASA-N Ala-Asp-Phe Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 MKZCBYZBCINNJN-DLOVCJGASA-N 0.000 description 4
- NWVVKQZOVSTDBQ-CIUDSAMLSA-N Ala-Glu-Arg Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O NWVVKQZOVSTDBQ-CIUDSAMLSA-N 0.000 description 4
- IPZQNYYAYVRKKK-FXQIFTODSA-N Ala-Pro-Ala Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O IPZQNYYAYVRKKK-FXQIFTODSA-N 0.000 description 4
- NCQMBSJGJMYKCK-ZLUOBGJFSA-N Ala-Ser-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O NCQMBSJGJMYKCK-ZLUOBGJFSA-N 0.000 description 4
- OBFTYSPXDRROQO-SRVKXCTJSA-N Arg-Gln-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CCCN=C(N)N OBFTYSPXDRROQO-SRVKXCTJSA-N 0.000 description 4
- ZATRYQNPUHGXCU-DTWKUNHWSA-N Arg-Gly-Pro Chemical compound C1C[C@@H](N(C1)C(=O)CNC(=O)[C@H](CCCN=C(N)N)N)C(=O)O ZATRYQNPUHGXCU-DTWKUNHWSA-N 0.000 description 4
- XQFLFQWOBXPMHW-NHCYSSNCSA-N Asp-Val-His Chemical compound N[C@@H](CC(=O)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)O XQFLFQWOBXPMHW-NHCYSSNCSA-N 0.000 description 4
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 4
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- MCAVASRGVBVPMX-FXQIFTODSA-N Gln-Glu-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O MCAVASRGVBVPMX-FXQIFTODSA-N 0.000 description 4
- HPJLZFTUUJKWAJ-JHEQGTHGSA-N Glu-Gly-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O HPJLZFTUUJKWAJ-JHEQGTHGSA-N 0.000 description 4
- NNCSJUBVFBDDLC-YUMQZZPRSA-N Gly-Leu-Ser Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O NNCSJUBVFBDDLC-YUMQZZPRSA-N 0.000 description 4
- KZTLOHBDLMIFSH-XVYDVKMFSA-N His-Ala-Asp Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(O)=O KZTLOHBDLMIFSH-XVYDVKMFSA-N 0.000 description 4
- YXBRCTXAEYSCHS-XVYDVKMFSA-N His-Ala-Cys Chemical compound C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N YXBRCTXAEYSCHS-XVYDVKMFSA-N 0.000 description 4
- BPOHQCZZSFBSON-KKUMJFAQSA-N His-Leu-His Chemical compound CC(C)C[C@H](NC(=O)[C@@H](N)Cc1cnc[nH]1)C(=O)N[C@@H](Cc1cnc[nH]1)C(O)=O BPOHQCZZSFBSON-KKUMJFAQSA-N 0.000 description 4
- CPELXLSAUQHCOX-UHFFFAOYSA-N Hydrogen bromide Chemical compound Br CPELXLSAUQHCOX-UHFFFAOYSA-N 0.000 description 4
- LJHGALIOHLRRQN-DCAQKATOSA-N Leu-Ala-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N LJHGALIOHLRRQN-DCAQKATOSA-N 0.000 description 4
- IBMVEYRWAWIOTN-RWMBFGLXSA-N Leu-Arg-Pro Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N1CCC[C@@H]1C(O)=O IBMVEYRWAWIOTN-RWMBFGLXSA-N 0.000 description 4
- 229930195725 Mannitol Natural products 0.000 description 4
- HDNOQCZWJGGHSS-VEVYYDQMSA-N Met-Asn-Thr Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O HDNOQCZWJGGHSS-VEVYYDQMSA-N 0.000 description 4
- GNRMAQSIROFNMI-IXOXFDKPSA-N Phe-Thr-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O GNRMAQSIROFNMI-IXOXFDKPSA-N 0.000 description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 4
- RVGRUAULSDPKGF-UHFFFAOYSA-N Poloxamer Chemical compound C1CO1.CC1CO1 RVGRUAULSDPKGF-UHFFFAOYSA-N 0.000 description 4
- HFZNNDWPHBRNPV-KZVJFYERSA-N Pro-Ala-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O HFZNNDWPHBRNPV-KZVJFYERSA-N 0.000 description 4
- ICTZKEXYDDZZFP-SRVKXCTJSA-N Pro-Arg-Pro Chemical compound N([C@@H](CCCN=C(N)N)C(=O)N1[C@@H](CCC1)C(O)=O)C(=O)[C@@H]1CCCN1 ICTZKEXYDDZZFP-SRVKXCTJSA-N 0.000 description 4
- GMJDSFYVTAMIBF-FXQIFTODSA-N Pro-Ser-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O GMJDSFYVTAMIBF-FXQIFTODSA-N 0.000 description 4
- CUXJENOFJXOSOZ-BIIVOSGPSA-N Ser-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CO)N)C(=O)O CUXJENOFJXOSOZ-BIIVOSGPSA-N 0.000 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- 210000001744 T-lymphocyte Anatomy 0.000 description 4
- 230000003187 abdominal effect Effects 0.000 description 4
- 230000000996 additive effect Effects 0.000 description 4
- 125000005907 alkyl ester group Chemical group 0.000 description 4
- 150000001408 amides Chemical group 0.000 description 4
- 125000000637 arginyl group Chemical group N[C@@H](CCCNC(N)=N)C(=O)* 0.000 description 4
- 210000004556 brain Anatomy 0.000 description 4
- 229910052799 carbon Inorganic materials 0.000 description 4
- 238000005336 cracking Methods 0.000 description 4
- 238000006073 displacement reaction Methods 0.000 description 4
- 150000002148 esters Chemical class 0.000 description 4
- 238000012637 gene transfection Methods 0.000 description 4
- 230000002068 genetic effect Effects 0.000 description 4
- 230000012447 hatching Effects 0.000 description 4
- 108010040030 histidinoalanine Proteins 0.000 description 4
- 108010028295 histidylhistidine Proteins 0.000 description 4
- 230000001900 immune effect Effects 0.000 description 4
- 230000005847 immunogenicity Effects 0.000 description 4
- 210000004962 mammalian cell Anatomy 0.000 description 4
- 239000000594 mannitol Substances 0.000 description 4
- 235000010355 mannitol Nutrition 0.000 description 4
- 108010051242 phenylalanylserine Proteins 0.000 description 4
- 229920000642 polymer Polymers 0.000 description 4
- 229960004063 propylene glycol Drugs 0.000 description 4
- 230000002441 reversible effect Effects 0.000 description 4
- 238000006467 substitution reaction Methods 0.000 description 4
- 239000000829 suppository Substances 0.000 description 4
- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical compound NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 description 4
- 230000001225 therapeutic effect Effects 0.000 description 4
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 4
- 238000001262 western blot Methods 0.000 description 4
- DSSYKIVIOFKYAU-XCBNKYQSSA-N (R)-camphor Chemical compound C1C[C@@]2(C)C(=O)C[C@@H]1C2(C)C DSSYKIVIOFKYAU-XCBNKYQSSA-N 0.000 description 3
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 3
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 3
- 108010016626 Dipeptides Proteins 0.000 description 3
- 108090000194 Dipeptidyl-peptidases and tripeptidyl-peptidases Proteins 0.000 description 3
- 102000003779 Dipeptidyl-peptidases and tripeptidyl-peptidases Human genes 0.000 description 3
- KGWDUNBJIMUFAP-KVVVOXFISA-N Ethanolamine Oleate Chemical compound NCCO.CCCCCCCC\C=C/CCCCCCCC(O)=O KGWDUNBJIMUFAP-KVVVOXFISA-N 0.000 description 3
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 3
- 101000908391 Homo sapiens Dipeptidyl peptidase 4 Proteins 0.000 description 3
- QYOGJYIRKACXEP-SLBDDTMCSA-N Ile-Asn-Trp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N QYOGJYIRKACXEP-SLBDDTMCSA-N 0.000 description 3
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 3
- AUBMZAMQCOYSIC-MNXVOIDGSA-N Leu-Ile-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(N)=O)C(O)=O AUBMZAMQCOYSIC-MNXVOIDGSA-N 0.000 description 3
- 102000000422 Matrix Metalloproteinase 3 Human genes 0.000 description 3
- 239000002202 Polyethylene glycol Substances 0.000 description 3
- 229920001213 Polysorbate 20 Polymers 0.000 description 3
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 3
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 3
- UUSQVWOVUYMLJA-PPCPHDFISA-N Thr-Lys-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O UUSQVWOVUYMLJA-PPCPHDFISA-N 0.000 description 3
- 108700005077 Viral Genes Proteins 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N acetic acid Substances CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 125000000217 alkyl group Chemical group 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 3
- 229960000846 camphor Drugs 0.000 description 3
- 150000001718 carbodiimides Chemical class 0.000 description 3
- 229910002091 carbon monoxide Inorganic materials 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 239000002299 complementary DNA Substances 0.000 description 3
- 229940090124 dipeptidyl peptidase 4 (dpp-4) inhibitors for blood glucose lowering Drugs 0.000 description 3
- 239000002552 dosage form Substances 0.000 description 3
- 231100000673 dose–response relationship Toxicity 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000000194 fatty acid Substances 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- UKVFVQPAANCXIL-FJVFSOETSA-N glp-1 (1-37) amide Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=CC=C1 UKVFVQPAANCXIL-FJVFSOETSA-N 0.000 description 3
- 229960002989 glutamic acid Drugs 0.000 description 3
- 239000004519 grease Substances 0.000 description 3
- 230000002949 hemolytic effect Effects 0.000 description 3
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 3
- 238000003119 immunoblot Methods 0.000 description 3
- 230000002779 inactivation Effects 0.000 description 3
- 239000003112 inhibitor Substances 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 238000003780 insertion Methods 0.000 description 3
- 230000037431 insertion Effects 0.000 description 3
- 210000000936 intestine Anatomy 0.000 description 3
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 230000007170 pathology Effects 0.000 description 3
- 239000013612 plasmid Substances 0.000 description 3
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 3
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 235000013772 propylene glycol Nutrition 0.000 description 3
- 102000005962 receptors Human genes 0.000 description 3
- 108020003175 receptors Proteins 0.000 description 3
- 230000008521 reorganization Effects 0.000 description 3
- 238000005204 segregation Methods 0.000 description 3
- APSBXTVYXVQYAB-UHFFFAOYSA-M sodium docusate Chemical compound [Na+].CCCCC(CC)COC(=O)CC(S([O-])(=O)=O)C(=O)OCC(CC)CCCC APSBXTVYXVQYAB-UHFFFAOYSA-M 0.000 description 3
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 3
- 230000004936 stimulating effect Effects 0.000 description 3
- 108091007196 stromelysin Proteins 0.000 description 3
- 238000007920 subcutaneous administration Methods 0.000 description 3
- 229940095064 tartrate Drugs 0.000 description 3
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 3
- 238000011144 upstream manufacturing Methods 0.000 description 3
- 238000012795 verification Methods 0.000 description 3
- BHQCQFFYRZLCQQ-UHFFFAOYSA-N (3alpha,5alpha,7alpha,12alpha)-3,7,12-trihydroxy-cholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 BHQCQFFYRZLCQQ-UHFFFAOYSA-N 0.000 description 2
- PORPENFLTBBHSG-MGBGTMOVSA-N 1,2-dihexadecanoyl-sn-glycerol-3-phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(O)=O)OC(=O)CCCCCCCCCCCCCCC PORPENFLTBBHSG-MGBGTMOVSA-N 0.000 description 2
- TZCPCKNHXULUIY-RGULYWFUSA-N 1,2-distearoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCCCCCCCCCCCC TZCPCKNHXULUIY-RGULYWFUSA-N 0.000 description 2
- NOGLQXZIGOQIBD-UHFFFAOYSA-N 1-(dimethylazaniumyl)propane-1-sulfonate Chemical compound CCC(N(C)C)S(O)(=O)=O NOGLQXZIGOQIBD-UHFFFAOYSA-N 0.000 description 2
- PAJPWUMXBYXFCZ-UHFFFAOYSA-N 1-aminocyclopropanecarboxylic acid Chemical compound OC(=O)C1(N)CC1 PAJPWUMXBYXFCZ-UHFFFAOYSA-N 0.000 description 2
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 2
- CFKMVGJGLGKFKI-UHFFFAOYSA-N 4-chloro-m-cresol Chemical compound CC1=CC(O)=CC=C1Cl CFKMVGJGLGKFKI-UHFFFAOYSA-N 0.000 description 2
- 229940090248 4-hydroxybenzoic acid Drugs 0.000 description 2
- DLFVBJFMPXGRIB-UHFFFAOYSA-N Acetamide Chemical compound CC(N)=O DLFVBJFMPXGRIB-UHFFFAOYSA-N 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 2
- HHGYNJRJIINWAK-FXQIFTODSA-N Ala-Ala-Arg Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N HHGYNJRJIINWAK-FXQIFTODSA-N 0.000 description 2
- 208000024827 Alzheimer disease Diseases 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 2
- PNHQRQTVBRDIEF-CIUDSAMLSA-N Asn-Leu-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(=O)N)N PNHQRQTVBRDIEF-CIUDSAMLSA-N 0.000 description 2
- RDLYUKRPEJERMM-XIRDDKMYSA-N Asn-Trp-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(C)C)C(O)=O RDLYUKRPEJERMM-XIRDDKMYSA-N 0.000 description 2
- NVFSJIXJZCDICF-SRVKXCTJSA-N Asp-Lys-Lys Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)O)N NVFSJIXJZCDICF-SRVKXCTJSA-N 0.000 description 2
- PCJOFZYFFMBZKC-PCBIJLKTSA-N Asp-Phe-Ile Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O PCJOFZYFFMBZKC-PCBIJLKTSA-N 0.000 description 2
- PWAIZUBWHRHYKS-MELADBBJSA-N Asp-Phe-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)NC(=O)[C@H](CC(=O)O)N)C(=O)O PWAIZUBWHRHYKS-MELADBBJSA-N 0.000 description 2
- QOJJMJKTMKNFEF-ZKWXMUAHSA-N Asp-Val-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC(O)=O QOJJMJKTMKNFEF-ZKWXMUAHSA-N 0.000 description 2
- 210000002237 B-cell of pancreatic islet Anatomy 0.000 description 2
- 239000004380 Cholic acid Substances 0.000 description 2
- 229920000858 Cyclodextrin Polymers 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 108010059378 Endopeptidases Proteins 0.000 description 2
- 102000005593 Endopeptidases Human genes 0.000 description 2
- IAYPIBMASNFSPL-UHFFFAOYSA-N Ethylene oxide Chemical compound C1CO1 IAYPIBMASNFSPL-UHFFFAOYSA-N 0.000 description 2
- QJCKNLPMTPXXEM-AUTRQRHGSA-N Glu-Glu-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CCC(O)=O QJCKNLPMTPXXEM-AUTRQRHGSA-N 0.000 description 2
- JZJGEKDPWVJOLD-QEWYBTABSA-N Glu-Phe-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JZJGEKDPWVJOLD-QEWYBTABSA-N 0.000 description 2
- ZWZWYGMENQVNFU-UHFFFAOYSA-N Glycerophosphorylserin Natural products OC(=O)C(N)COP(O)(=O)OCC(O)CO ZWZWYGMENQVNFU-UHFFFAOYSA-N 0.000 description 2
- 241000270431 Heloderma suspectum Species 0.000 description 2
- 101000684208 Homo sapiens Prolyl endopeptidase FAP Proteins 0.000 description 2
- JRYQSFOFUFXPTB-RWRJDSDZSA-N Ile-Gln-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N JRYQSFOFUFXPTB-RWRJDSDZSA-N 0.000 description 2
- 206010022489 Insulin Resistance Diseases 0.000 description 2
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 2
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- XBBKIIGCUMBKCO-JXUBOQSCSA-N Leu-Ala-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XBBKIIGCUMBKCO-JXUBOQSCSA-N 0.000 description 2
- NCZIQZYZPUPMKY-PPCPHDFISA-N Lys-Ile-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O NCZIQZYZPUPMKY-PPCPHDFISA-N 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- DBMLDOWSVHMQQN-XGEHTFHBSA-N Met-Ser-Thr Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O DBMLDOWSVHMQQN-XGEHTFHBSA-N 0.000 description 2
- 101100395023 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) his-7 gene Proteins 0.000 description 2
- JWBLQDDHSDGEGR-DRZSPHRISA-N Phe-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 JWBLQDDHSDGEGR-DRZSPHRISA-N 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- 239000004372 Polyvinyl alcohol Substances 0.000 description 2
- HCBIBCJNVBAKAB-UHFFFAOYSA-N Procaine hydrochloride Chemical compound Cl.CCN(CC)CCOC(=O)C1=CC=C(N)C=C1 HCBIBCJNVBAKAB-UHFFFAOYSA-N 0.000 description 2
- 108010058003 Proglucagon Proteins 0.000 description 2
- ATUOYWHBWRKTHZ-UHFFFAOYSA-N Propane Chemical compound CCC ATUOYWHBWRKTHZ-UHFFFAOYSA-N 0.000 description 2
- RADKZDMFGJYCBB-UHFFFAOYSA-N Pyridoxal Chemical compound CC1=NC=C(CO)C(C=O)=C1O RADKZDMFGJYCBB-UHFFFAOYSA-N 0.000 description 2
- SMWDFEZZVXVKRB-UHFFFAOYSA-N Quinoline Chemical compound N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- LYMVXFSTACVOLP-ZFWWWQNUSA-N Trp-Leu Chemical compound C1=CC=C2C(C[C@H]([NH3+])C(=O)N[C@@H](CC(C)C)C([O-])=O)=CNC2=C1 LYMVXFSTACVOLP-ZFWWWQNUSA-N 0.000 description 2
- WMBFONUKQXGLMU-WDSOQIARSA-N Trp-Leu-Val Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N WMBFONUKQXGLMU-WDSOQIARSA-N 0.000 description 2
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 2
- 239000006035 Tryptophane Substances 0.000 description 2
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 125000002252 acyl group Chemical group 0.000 description 2
- UDMBCSSLTHHNCD-KQYNXXCUSA-N adenosine 5'-monophosphate Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(O)=O)[C@@H](O)[C@H]1O UDMBCSSLTHHNCD-KQYNXXCUSA-N 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 125000003545 alkoxy group Chemical group 0.000 description 2
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 2
- 125000003277 amino group Chemical group 0.000 description 2
- 206010002026 amyotrophic lateral sclerosis Diseases 0.000 description 2
- 230000008485 antagonism Effects 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 125000000613 asparagine group Chemical group N[C@@H](CC(N)=O)C(=O)* 0.000 description 2
- 235000003704 aspartic acid Nutrition 0.000 description 2
- 229960005261 aspartic acid Drugs 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 125000004429 atom Chemical group 0.000 description 2
- 210000000227 basophil cell of anterior lobe of hypophysis Anatomy 0.000 description 2
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- QARVLSVVCXYDNA-UHFFFAOYSA-N bromobenzene Chemical compound BrC1=CC=CC=C1 QARVLSVVCXYDNA-UHFFFAOYSA-N 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- DWKPZOZZBLWFJX-UHFFFAOYSA-L calcium;1,4-bis(2-ethylhexoxy)-1,4-dioxobutane-2-sulfonate Chemical compound [Ca+2].CCCCC(CC)COC(=O)CC(S([O-])(=O)=O)C(=O)OCC(CC)CCCC.CCCCC(CC)COC(=O)CC(S([O-])(=O)=O)C(=O)OCC(CC)CCCC DWKPZOZZBLWFJX-UHFFFAOYSA-L 0.000 description 2
- 230000023852 carbohydrate metabolic process Effects 0.000 description 2
- 235000021256 carbohydrate metabolism Nutrition 0.000 description 2
- 239000004359 castor oil Substances 0.000 description 2
- 235000019438 castor oil Nutrition 0.000 description 2
- 230000003915 cell function Effects 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 210000003169 central nervous system Anatomy 0.000 description 2
- 239000000460 chlorine Substances 0.000 description 2
- 229910052801 chlorine Inorganic materials 0.000 description 2
- 235000019416 cholic acid Nutrition 0.000 description 2
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 2
- 229960002471 cholic acid Drugs 0.000 description 2
- 229960001231 choline Drugs 0.000 description 2
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 230000021615 conjugation Effects 0.000 description 2
- 125000004122 cyclic group Chemical group 0.000 description 2
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 2
- 238000013016 damping Methods 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- KXGVEGMKQFWNSR-UHFFFAOYSA-N deoxycholic acid Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 KXGVEGMKQFWNSR-UHFFFAOYSA-N 0.000 description 2
- 238000001212 derivatisation Methods 0.000 description 2
- 235000019329 dioctyl sodium sulphosuccinate Nutrition 0.000 description 2
- 229960000878 docusate sodium Drugs 0.000 description 2
- LQZZUXJYWNFBMV-UHFFFAOYSA-N dodecan-1-ol Chemical compound CCCCCCCCCCCCO LQZZUXJYWNFBMV-UHFFFAOYSA-N 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- HDERJYVLTPVNRI-UHFFFAOYSA-N ethene;ethenyl acetate Chemical compound C=C.CC(=O)OC=C HDERJYVLTPVNRI-UHFFFAOYSA-N 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000037406 food intake Effects 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- FBPFZTCFMRRESA-GUCUJZIJSA-N galactitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-GUCUJZIJSA-N 0.000 description 2
- 229940121355 glucagon like peptide 2 (glp-2) analogues Drugs 0.000 description 2
- 125000000404 glutamine group Chemical group N[C@@H](CCC(N)=O)C(=O)* 0.000 description 2
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 description 2
- 108010074027 glycyl-seryl-phenylalanine Proteins 0.000 description 2
- YMAWOPBAYDPSLA-UHFFFAOYSA-N glycylglycine Chemical compound [NH3+]CC(=O)NCC([O-])=O YMAWOPBAYDPSLA-UHFFFAOYSA-N 0.000 description 2
- HHLFWLYXYJOTON-UHFFFAOYSA-N glyoxylic acid Chemical compound OC(=O)C=O HHLFWLYXYJOTON-UHFFFAOYSA-N 0.000 description 2
- 229910052736 halogen Inorganic materials 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 235000020256 human milk Nutrition 0.000 description 2
- 210000004251 human milk Anatomy 0.000 description 2
- 229910000042 hydrogen bromide Inorganic materials 0.000 description 2
- 201000001421 hyperglycemia Diseases 0.000 description 2
- CDAISMWEOUEBRE-UHFFFAOYSA-N inositol Chemical compound OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 2
- 230000010354 integration Effects 0.000 description 2
- 238000010253 intravenous injection Methods 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical class CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 2
- 108091005601 modified peptides Proteins 0.000 description 2
- 239000000178 monomer Substances 0.000 description 2
- 230000001537 neural effect Effects 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 230000000050 nutritive effect Effects 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 235000006408 oxalic acid Nutrition 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- OJUGVDODNPJEEC-UHFFFAOYSA-N phenylglyoxal Chemical compound O=CC(=O)C1=CC=CC=C1 OJUGVDODNPJEEC-UHFFFAOYSA-N 0.000 description 2
- 229960000502 poloxamer Drugs 0.000 description 2
- 229920001983 poloxamer Polymers 0.000 description 2
- 229920000728 polyester Polymers 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 229920002451 polyvinyl alcohol Polymers 0.000 description 2
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 2
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 2
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 2
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 2
- CACRHRQTJDKAPJ-UHFFFAOYSA-M potassium;1,4-bis(2-ethylhexoxy)-1,4-dioxobutane-2-sulfonate Chemical compound [K+].CCCCC(CC)COC(=O)CC(S([O-])(=O)=O)C(=O)OCC(CC)CCCC CACRHRQTJDKAPJ-UHFFFAOYSA-M 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 108010070643 prolylglutamic acid Proteins 0.000 description 2
- 239000001294 propane Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- NGVDGCNFYWLIFO-UHFFFAOYSA-N pyridoxal 5'-phosphate Chemical compound CC1=NC=C(COP(O)(O)=O)C(C=O)=C1O NGVDGCNFYWLIFO-UHFFFAOYSA-N 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000036186 satiety Effects 0.000 description 2
- 235000019627 satiety Nutrition 0.000 description 2
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 230000003248 secreting effect Effects 0.000 description 2
- 210000002955 secretory cell Anatomy 0.000 description 2
- 239000003352 sequestering agent Substances 0.000 description 2
- 238000004088 simulation Methods 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 210000004988 splenocyte Anatomy 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 150000003462 sulfoxides Chemical class 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 229960003080 taurine Drugs 0.000 description 2
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 2
- GETQZCLCWQTVFV-UHFFFAOYSA-N trimethylamine Chemical compound CN(C)C GETQZCLCWQTVFV-UHFFFAOYSA-N 0.000 description 2
- 229960004799 tryptophan Drugs 0.000 description 2
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 2
- 230000003827 upregulation Effects 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- USMNOWBWPHYOEA-XKSSXDPKSA-N (+)-beta-thujone Chemical compound O=C([C@H]1C)C[C@@]2(C(C)C)[C@@H]1C2 USMNOWBWPHYOEA-XKSSXDPKSA-N 0.000 description 1
- WWUZIQQURGPMPG-UHFFFAOYSA-N (-)-D-erythro-Sphingosine Natural products CCCCCCCCCCCCCC=CC(O)C(N)CO WWUZIQQURGPMPG-UHFFFAOYSA-N 0.000 description 1
- MEIRRNXMZYDVDW-MQQKCMAXSA-N (2E,4E)-2,4-hexadien-1-ol Chemical compound C\C=C\C=C\CO MEIRRNXMZYDVDW-MQQKCMAXSA-N 0.000 description 1
- SNRCMWGPCHPRBP-YFKPBYRVSA-N (2s)-2-(hydroxyamino)-3-(1h-imidazol-5-yl)propanoic acid Chemical compound ON[C@H](C(O)=O)CC1=CNC=N1 SNRCMWGPCHPRBP-YFKPBYRVSA-N 0.000 description 1
- MSECZMWQBBVGEN-LURJTMIESA-N (2s)-2-azaniumyl-4-(1h-imidazol-5-yl)butanoate Chemical compound OC(=O)[C@@H](N)CCC1=CN=CN1 MSECZMWQBBVGEN-LURJTMIESA-N 0.000 description 1
- ASWBNKHCZGQVJV-UHFFFAOYSA-N (3-hexadecanoyloxy-2-hydroxypropyl) 2-(trimethylazaniumyl)ethyl phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(O)COP([O-])(=O)OCC[N+](C)(C)C ASWBNKHCZGQVJV-UHFFFAOYSA-N 0.000 description 1
- PYIDGJJWBIBVIA-IHAUNJBESA-N (3r,4s,5s,6r)-2-dodecoxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound CCCCCCCCCCCCOC1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O PYIDGJJWBIBVIA-IHAUNJBESA-N 0.000 description 1
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 1
- MXOAEAUPQDYUQM-QMMMGPOBSA-N (S)-chlorphenesin Chemical compound OC[C@H](O)COC1=CC=C(Cl)C=C1 MXOAEAUPQDYUQM-QMMMGPOBSA-N 0.000 description 1
- 230000006269 (delayed) early viral mRNA transcription Effects 0.000 description 1
- LDVVTQMJQSCDMK-UHFFFAOYSA-N 1,3-dihydroxypropan-2-yl formate Chemical compound OCC(CO)OC=O LDVVTQMJQSCDMK-UHFFFAOYSA-N 0.000 description 1
- VNBFUGOVQMFIRN-UHFFFAOYSA-N 1-chlorobutan-2-ol Chemical compound CCC(O)CCl VNBFUGOVQMFIRN-UHFFFAOYSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- LDGWQMRUWMSZIU-LQDDAWAPSA-M 2,3-bis[(z)-octadec-9-enoxy]propyl-trimethylazanium;chloride Chemical compound [Cl-].CCCCCCCC\C=C/CCCCCCCCOCC(C[N+](C)(C)C)OCCCCCCCC\C=C/CCCCCCCC LDGWQMRUWMSZIU-LQDDAWAPSA-M 0.000 description 1
- ZZXDRXVIRVJQBT-UHFFFAOYSA-N 2,3-dimethylbenzenesulfonic acid Chemical class CC1=CC=CC(S(O)(=O)=O)=C1C ZZXDRXVIRVJQBT-UHFFFAOYSA-N 0.000 description 1
- NHJVRSWLHSJWIN-UHFFFAOYSA-N 2,4,6-trinitrobenzenesulfonic acid Chemical compound OS(=O)(=O)C1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O NHJVRSWLHSJWIN-UHFFFAOYSA-N 0.000 description 1
- NMPVEAUIHMEAQP-UHFFFAOYSA-N 2-Bromoacetaldehyde Chemical compound BrCC=O NMPVEAUIHMEAQP-UHFFFAOYSA-N 0.000 description 1
- IEQAICDLOKRSRL-UHFFFAOYSA-N 2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2-dodecoxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethanol Chemical compound CCCCCCCCCCCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCO IEQAICDLOKRSRL-UHFFFAOYSA-N 0.000 description 1
- UXFQFBNBSPQBJW-UHFFFAOYSA-N 2-amino-2-methylpropane-1,3-diol Chemical compound OCC(N)(C)CO UXFQFBNBSPQBJW-UHFFFAOYSA-N 0.000 description 1
- CRTWOYOCILXSSX-UHFFFAOYSA-N 2-bromo-3-(1h-imidazol-5-yl)propanoic acid Chemical compound OC(=O)C(Br)CC1=CN=CN1 CRTWOYOCILXSSX-UHFFFAOYSA-N 0.000 description 1
- BSKHPKMHTQYZBB-UHFFFAOYSA-N 2-methylpyridine Chemical compound CC1=CC=CC=N1 BSKHPKMHTQYZBB-UHFFFAOYSA-N 0.000 description 1
- SLAMLWHELXOEJZ-UHFFFAOYSA-M 2-nitrobenzoate Chemical compound [O-]C(=O)C1=CC=CC=C1[N+]([O-])=O SLAMLWHELXOEJZ-UHFFFAOYSA-M 0.000 description 1
- QCDWFXQBSFUVSP-UHFFFAOYSA-N 2-phenoxyethanol Chemical compound OCCOC1=CC=CC=C1 QCDWFXQBSFUVSP-UHFFFAOYSA-N 0.000 description 1
- OFJWFSNDPCAWDK-UHFFFAOYSA-N 2-phenylbutyric acid Chemical class CCC(C(O)=O)C1=CC=CC=C1 OFJWFSNDPCAWDK-UHFFFAOYSA-N 0.000 description 1
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 1
- GSWYUZQBLVUEPH-UHFFFAOYSA-N 3-(azaniumylmethyl)benzoate Chemical compound NCC1=CC=CC(C(O)=O)=C1 GSWYUZQBLVUEPH-UHFFFAOYSA-N 0.000 description 1
- PQDKQNSTGLBIAT-UHFFFAOYSA-N 3-methylpyridin-1-ium-2-sulfonate Chemical compound CC1=CC=CN=C1S(O)(=O)=O PQDKQNSTGLBIAT-UHFFFAOYSA-N 0.000 description 1
- WZBDCLMTYAGWKV-UHFFFAOYSA-N 3-nitropyridine-2-sulfonic acid Chemical compound OS(=O)(=O)C1=NC=CC=C1[N+]([O-])=O WZBDCLMTYAGWKV-UHFFFAOYSA-N 0.000 description 1
- PXACTUVBBMDKRW-UHFFFAOYSA-N 4-bromobenzenesulfonic acid Chemical compound OS(=O)(=O)C1=CC=C(Br)C=C1 PXACTUVBBMDKRW-UHFFFAOYSA-N 0.000 description 1
- BBMFSGOFUHEVNP-UHFFFAOYSA-N 4-hydroxy-2-methylbenzoic acid Chemical compound CC1=CC(O)=CC=C1C(O)=O BBMFSGOFUHEVNP-UHFFFAOYSA-N 0.000 description 1
- SJZRECIVHVDYJC-UHFFFAOYSA-N 4-hydroxybutyric acid Chemical class OCCCC(O)=O SJZRECIVHVDYJC-UHFFFAOYSA-N 0.000 description 1
- IJJWOSAXNHWBPR-HUBLWGQQSA-N 5-[(3as,4s,6ar)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]-n-(6-hydrazinyl-6-oxohexyl)pentanamide Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)NCCCCCC(=O)NN)SC[C@@H]21 IJJWOSAXNHWBPR-HUBLWGQQSA-N 0.000 description 1
- WOVKYSAHUYNSMH-RRKCRQDMSA-N 5-bromodeoxyuridine Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(Br)=C1 WOVKYSAHUYNSMH-RRKCRQDMSA-N 0.000 description 1
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-M Acrylate Chemical compound [O-]C(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-M 0.000 description 1
- MAZZQZWCCYJQGZ-GUBZILKMSA-N Ala-Pro-Arg Chemical compound [H]N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(O)=O MAZZQZWCCYJQGZ-GUBZILKMSA-N 0.000 description 1
- 208000011403 Alexander disease Diseases 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 102000004400 Aminopeptidases Human genes 0.000 description 1
- 108090000915 Aminopeptidases Proteins 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical class N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 102000014303 Amyloid beta-Protein Precursor Human genes 0.000 description 1
- 108010079054 Amyloid beta-Protein Precursor Proteins 0.000 description 1
- IASNWHAGGYTEKX-IUCAKERBSA-N Arg-Arg-Gly Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)NCC(O)=O IASNWHAGGYTEKX-IUCAKERBSA-N 0.000 description 1
- CIBWFJFMOBIFTE-CIUDSAMLSA-N Asn-Arg-Gln Chemical compound C(C[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N)CN=C(N)N CIBWFJFMOBIFTE-CIUDSAMLSA-N 0.000 description 1
- QCVXMEHGFUMKCO-YUMQZZPRSA-N Asp-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC(O)=O QCVXMEHGFUMKCO-YUMQZZPRSA-N 0.000 description 1
- FAUPLTGRUBTXNU-FXQIFTODSA-N Asp-Pro-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O FAUPLTGRUBTXNU-FXQIFTODSA-N 0.000 description 1
- 206010003591 Ataxia Diseases 0.000 description 1
- 206010003594 Ataxia telangiectasia Diseases 0.000 description 1
- 238000011725 BALB/c mouse Methods 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- WVDDGKGOMKODPV-UHFFFAOYSA-N Benzyl alcohol Chemical compound OCC1=CC=CC=C1 WVDDGKGOMKODPV-UHFFFAOYSA-N 0.000 description 1
- 241000212384 Bifora Species 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- CPELXLSAUQHCOX-UHFFFAOYSA-M Bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 description 1
- LVDKZNITIUWNER-UHFFFAOYSA-N Bronopol Chemical compound OCC(Br)(CO)[N+]([O-])=O LVDKZNITIUWNER-UHFFFAOYSA-N 0.000 description 1
- COVZYZSDYWQREU-UHFFFAOYSA-N Busulfan Chemical compound CS(=O)(=O)OCCCCOS(C)(=O)=O COVZYZSDYWQREU-UHFFFAOYSA-N 0.000 description 1
- YNNKVAPENUJLEK-UHFFFAOYSA-L C(=O)[O-].C1=CC=CC=C1.Cl[Hg+] Chemical compound C(=O)[O-].C1=CC=CC=C1.Cl[Hg+] YNNKVAPENUJLEK-UHFFFAOYSA-L 0.000 description 1
- YDNKGFDKKRUKPY-JHOUSYSJSA-N C16 ceramide Natural products CCCCCCCCCCCCCCCC(=O)N[C@@H](CO)[C@H](O)C=CCCCCCCCCCCCCC YDNKGFDKKRUKPY-JHOUSYSJSA-N 0.000 description 1
- OBMZMSLWNNWEJA-XNCRXQDQSA-N C1=CC=2C(C[C@@H]3NC(=O)[C@@H](NC(=O)[C@H](NC(=O)N(CC#CCN(CCCC[C@H](NC(=O)[C@@H](CC4=CC=CC=C4)NC3=O)C(=O)N)CC=C)NC(=O)[C@@H](N)C)CC3=CNC4=C3C=CC=C4)C)=CNC=2C=C1 Chemical compound C1=CC=2C(C[C@@H]3NC(=O)[C@@H](NC(=O)[C@H](NC(=O)N(CC#CCN(CCCC[C@H](NC(=O)[C@@H](CC4=CC=CC=C4)NC3=O)C(=O)N)CC=C)NC(=O)[C@@H](N)C)CC3=CNC4=C3C=CC=C4)C)=CNC=2C=C1 OBMZMSLWNNWEJA-XNCRXQDQSA-N 0.000 description 1
- 101100335894 Caenorhabditis elegans gly-8 gene Proteins 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- KXDHJXZQYSOELW-UHFFFAOYSA-N Carbamic acid Chemical compound NC(O)=O KXDHJXZQYSOELW-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 1
- GHXZTYHSJHQHIJ-UHFFFAOYSA-N Chlorhexidine Chemical compound C=1C=C(Cl)C=CC=1NC(N)=NC(N)=NCCCCCCN=C(N)N=C(N)NC1=CC=C(Cl)C=C1 GHXZTYHSJHQHIJ-UHFFFAOYSA-N 0.000 description 1
- 235000007516 Chrysanthemum Nutrition 0.000 description 1
- 244000189548 Chrysanthemum x morifolium Species 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 108010062580 Concanavalin A Proteins 0.000 description 1
- 208000020406 Creutzfeldt Jacob disease Diseases 0.000 description 1
- 208000003407 Creutzfeldt-Jakob Syndrome Diseases 0.000 description 1
- 208000010859 Creutzfeldt-Jakob disease Diseases 0.000 description 1
- HEBKCHPVOIAQTA-QWWZWVQMSA-N D-arabinitol Chemical compound OC[C@@H](O)C(O)[C@H](O)CO HEBKCHPVOIAQTA-QWWZWVQMSA-N 0.000 description 1
- ZDXPYRJPNDTMRX-GSVOUGTGSA-N D-glutamine Chemical compound OC(=O)[C@H](N)CCC(N)=O ZDXPYRJPNDTMRX-GSVOUGTGSA-N 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- MEQISPGLXJPPIG-UHFFFAOYSA-N FC(C(C)=O)(F)F.[Br] Chemical compound FC(C(C)=O)(F)F.[Br] MEQISPGLXJPPIG-UHFFFAOYSA-N 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 102100024785 Fibroblast growth factor 2 Human genes 0.000 description 1
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 description 1
- BDAGIHXWWSANSR-UHFFFAOYSA-M Formate Chemical compound [O-]C=O BDAGIHXWWSANSR-UHFFFAOYSA-M 0.000 description 1
- 201000011240 Frontotemporal dementia Diseases 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 108010004460 Gastric Inhibitory Polypeptide Proteins 0.000 description 1
- 102100039994 Gastric inhibitory polypeptide Human genes 0.000 description 1
- 108700028146 Genetic Enhancer Elements Proteins 0.000 description 1
- 101710173678 Glucagon-5 Proteins 0.000 description 1
- 102000007446 Glucagon-Like Peptide-1 Receptor Human genes 0.000 description 1
- 108010086246 Glucagon-Like Peptide-1 Receptor Proteins 0.000 description 1
- 102400000324 Glucagon-like peptide 1(7-37) Human genes 0.000 description 1
- 229920001503 Glucan Polymers 0.000 description 1
- YGHSQRJSHKYUJY-SCZZXKLOSA-N Gly-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN YGHSQRJSHKYUJY-SCZZXKLOSA-N 0.000 description 1
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 description 1
- 108010008488 Glycylglycine Proteins 0.000 description 1
- 229920003114 HPC-L Polymers 0.000 description 1
- 229920003115 HPC-SL Polymers 0.000 description 1
- BIAKMWKJMQLZOJ-ZKWXMUAHSA-N His-Ala-Ala Chemical compound C[C@H](NC(=O)[C@H](C)NC(=O)[C@@H](N)Cc1cnc[nH]1)C(O)=O BIAKMWKJMQLZOJ-ZKWXMUAHSA-N 0.000 description 1
- HAPWZEVRQYGLSG-IUCAKERBSA-N His-Gly-Glu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O HAPWZEVRQYGLSG-IUCAKERBSA-N 0.000 description 1
- PMWSGVRIMIFXQH-KKUMJFAQSA-N His-His-Leu Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@@H](N)CC=1NC=NC=1)C1=CN=CN1 PMWSGVRIMIFXQH-KKUMJFAQSA-N 0.000 description 1
- 101500028773 Homo sapiens Glucagon-like peptide 1(7-37) Proteins 0.000 description 1
- 229920002153 Hydroxypropyl cellulose Polymers 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 208000009164 Islet Cell Adenoma Diseases 0.000 description 1
- 206010023379 Ketoacidosis Diseases 0.000 description 1
- 208000007976 Ketosis Diseases 0.000 description 1
- LKDRXBCSQODPBY-AMVSKUEXSA-N L-(-)-Sorbose Chemical compound OCC1(O)OC[C@H](O)[C@@H](O)[C@@H]1O LKDRXBCSQODPBY-AMVSKUEXSA-N 0.000 description 1
- AHLPHDHHMVZTML-BYPYZUCNSA-N L-Ornithine Chemical compound NCCC[C@H](N)C(O)=O AHLPHDHHMVZTML-BYPYZUCNSA-N 0.000 description 1
- 125000000570 L-alpha-aspartyl group Chemical group [H]OC(=O)C([H])([H])[C@]([H])(N([H])[H])C(*)=O 0.000 description 1
- 125000002059 L-arginyl group Chemical group O=C([*])[C@](N([H])[H])([H])C([H])([H])C([H])([H])C([H])([H])N([H])C(=N[H])N([H])[H] 0.000 description 1
- 125000002061 L-isoleucyl group Chemical group [H]N([H])[C@]([H])(C(=O)[*])[C@](C([H])([H])[H])([H])C(C([H])([H])[H])([H])[H] 0.000 description 1
- 125000003440 L-leucyl group Chemical group O=C([*])[C@](N([H])[H])([H])C([H])([H])C(C([H])([H])[H])([H])C([H])([H])[H] 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 125000002435 L-phenylalanyl group Chemical group O=C([*])[C@](N([H])[H])([H])C([H])([H])C1=C([H])C([H])=C([H])C([H])=C1[H] 0.000 description 1
- 125000003798 L-tyrosyl group Chemical group [H]N([H])[C@]([H])(C(=O)[*])C([H])([H])C1=C([H])C([H])=C(O[H])C([H])=C1[H] 0.000 description 1
- CHJKEDSZNSONPS-DCAQKATOSA-N Leu-Pro-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O CHJKEDSZNSONPS-DCAQKATOSA-N 0.000 description 1
- OFOBLEOULBTSOW-UHFFFAOYSA-L Malonate Chemical compound [O-]C(=O)CC([O-])=O OFOBLEOULBTSOW-UHFFFAOYSA-L 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 108010076502 Matrix Metalloproteinase 11 Proteins 0.000 description 1
- 241000511739 Melampyrum Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- VIHYIVKEECZGOU-UHFFFAOYSA-N N-acetylimidazole Chemical compound CC(=O)N1C=CN=C1 VIHYIVKEECZGOU-UHFFFAOYSA-N 0.000 description 1
- CRJGESKKUOMBCT-VQTJNVASSA-N N-acetylsphinganine Chemical compound CCCCCCCCCCCCCCC[C@@H](O)[C@H](CO)NC(C)=O CRJGESKKUOMBCT-VQTJNVASSA-N 0.000 description 1
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 1
- 241001391926 Neea Species 0.000 description 1
- BZQFBWGGLXLEPQ-UHFFFAOYSA-N O-phosphoryl-L-serine Natural products OC(=O)C(N)COP(O)(O)=O BZQFBWGGLXLEPQ-UHFFFAOYSA-N 0.000 description 1
- 208000008589 Obesity Diseases 0.000 description 1
- 239000005642 Oleic acid Substances 0.000 description 1
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 1
- 101500016420 Oncorhynchus mykiss Glucagon-like peptide 1-1 Proteins 0.000 description 1
- AHLPHDHHMVZTML-UHFFFAOYSA-N Orn-delta-NH2 Natural products NCCCC(N)C(O)=O AHLPHDHHMVZTML-UHFFFAOYSA-N 0.000 description 1
- UTJLXEIPEHZYQJ-UHFFFAOYSA-N Ornithine Natural products OC(=O)C(C)CCCN UTJLXEIPEHZYQJ-UHFFFAOYSA-N 0.000 description 1
- 206010033307 Overweight Diseases 0.000 description 1
- 208000018737 Parkinson disease Diseases 0.000 description 1
- 101710176384 Peptide 1 Proteins 0.000 description 1
- 208000000609 Pick Disease of the Brain Diseases 0.000 description 1
- 229920002562 Polyethylene Glycol 3350 Polymers 0.000 description 1
- 229920002565 Polyethylene Glycol 400 Polymers 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- CGBYDGAJHSOGFQ-LPEHRKFASA-N Pro-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@@H]2CCCN2 CGBYDGAJHSOGFQ-LPEHRKFASA-N 0.000 description 1
- XQLBWXHVZVBNJM-FXQIFTODSA-N Pro-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1 XQLBWXHVZVBNJM-FXQIFTODSA-N 0.000 description 1
- XUSDDSLCRPUKLP-QXEWZRGKSA-N Pro-Asp-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H]1CCCN1 XUSDDSLCRPUKLP-QXEWZRGKSA-N 0.000 description 1
- JLMZKEQFMVORMA-SRVKXCTJSA-N Pro-Pro-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 JLMZKEQFMVORMA-SRVKXCTJSA-N 0.000 description 1
- 102000035554 Proglucagon Human genes 0.000 description 1
- GOOHAUXETOMSMM-UHFFFAOYSA-N Propylene oxide Chemical compound CC1CO1 GOOHAUXETOMSMM-UHFFFAOYSA-N 0.000 description 1
- 102000007327 Protamines Human genes 0.000 description 1
- 108010007568 Protamines Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 description 1
- 102000001253 Protein Kinase Human genes 0.000 description 1
- 239000004373 Pullulan Substances 0.000 description 1
- 229920001218 Pullulan Polymers 0.000 description 1
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 1
- 208000001647 Renal Insufficiency Diseases 0.000 description 1
- 108010039491 Ricin Proteins 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- WINXNKPZLFISPD-UHFFFAOYSA-M Saccharin sodium Chemical compound [Na+].C1=CC=C2C(=O)[N-]S(=O)(=O)C2=C1 WINXNKPZLFISPD-UHFFFAOYSA-M 0.000 description 1
- 208000021811 Sandhoff disease Diseases 0.000 description 1
- FIXILCYTSAUERA-FXQIFTODSA-N Ser-Ala-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FIXILCYTSAUERA-FXQIFTODSA-N 0.000 description 1
- QYSFWUIXDFJUDW-DCAQKATOSA-N Ser-Leu-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O QYSFWUIXDFJUDW-DCAQKATOSA-N 0.000 description 1
- ADJDNJCSPNFFPI-FXQIFTODSA-N Ser-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CO ADJDNJCSPNFFPI-FXQIFTODSA-N 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 239000004288 Sodium dehydroacetate Substances 0.000 description 1
- 239000004141 Sodium laurylsulphate Substances 0.000 description 1
- 102000017303 Stromelysin-3 Human genes 0.000 description 1
- NYTOUQBROMCLBJ-UHFFFAOYSA-N Tetranitromethane Chemical compound [O-][N+](=O)C([N+]([O-])=O)([N+]([O-])=O)[N+]([O-])=O NYTOUQBROMCLBJ-UHFFFAOYSA-N 0.000 description 1
- VXMHQKHDKCATDV-VEVYYDQMSA-N Thr-Asp-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O VXMHQKHDKCATDV-VEVYYDQMSA-N 0.000 description 1
- VUVCRYXYUUPGSB-GLLZPBPUSA-N Thr-Gln-Glu Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N)O VUVCRYXYUUPGSB-GLLZPBPUSA-N 0.000 description 1
- 102000003929 Transaminases Human genes 0.000 description 1
- 108090000340 Transaminases Proteins 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- BNOODXBBXFZASF-UHFFFAOYSA-N [Na].[S] Chemical compound [Na].[S] BNOODXBBXFZASF-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- LPQOADBMXVRBNX-UHFFFAOYSA-N ac1ldcw0 Chemical compound Cl.C1CN(C)CCN1C1=C(F)C=C2C(=O)C(C(O)=O)=CN3CCSC1=C32 LPQOADBMXVRBNX-UHFFFAOYSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 1
- BYNPVFHVZRNRQD-UHFFFAOYSA-N acetic acid;benzene Chemical compound CC(O)=O.C1=CC=CC=C1 BYNPVFHVZRNRQD-UHFFFAOYSA-N 0.000 description 1
- YRKCREAYFQTBPV-UHFFFAOYSA-N acetylacetone Chemical compound CC(=O)CC(C)=O YRKCREAYFQTBPV-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 102000030621 adenylate cyclase Human genes 0.000 description 1
- 108060000200 adenylate cyclase Proteins 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- 150000001447 alkali salts Chemical class 0.000 description 1
- 150000001336 alkenes Chemical class 0.000 description 1
- 230000000735 allogeneic effect Effects 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- WQZGKKKJIJFFOK-PQMKYFCFSA-N alpha-D-mannose Chemical compound OC[C@H]1O[C@H](O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-PQMKYFCFSA-N 0.000 description 1
- SRBFZHDQGSBBOR-LECHCGJUSA-N alpha-D-xylose Chemical compound O[C@@H]1CO[C@H](O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-LECHCGJUSA-N 0.000 description 1
- QWCKQJZIFLGMSD-UHFFFAOYSA-N alpha-aminobutyric acid Chemical compound CCC(N)C(O)=O QWCKQJZIFLGMSD-UHFFFAOYSA-N 0.000 description 1
- 150000001409 amidines Chemical class 0.000 description 1
- 230000006229 amino acid addition Effects 0.000 description 1
- 150000003862 amino acid derivatives Chemical class 0.000 description 1
- 150000001414 amino alcohols Chemical class 0.000 description 1
- 229940124277 aminobutyric acid Drugs 0.000 description 1
- 125000004202 aminomethyl group Chemical group [H]N([H])C([H])([H])* 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 230000002424 anti-apoptotic effect Effects 0.000 description 1
- 230000003178 anti-diabetic effect Effects 0.000 description 1
- 230000000636 anti-proteolytic effect Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 108010062796 arginyllysine Proteins 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 238000011888 autopsy Methods 0.000 description 1
- OGBUMNBNEWYMNJ-UHFFFAOYSA-N batilol Chemical class CCCCCCCCCCCCCCCCCCOCC(O)CO OGBUMNBNEWYMNJ-UHFFFAOYSA-N 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 229960001950 benzethonium chloride Drugs 0.000 description 1
- UREZNYTWGJKWBI-UHFFFAOYSA-M benzethonium chloride Chemical compound [Cl-].C1=CC(C(C)(C)CC(C)(C)C)=CC=C1OCCOCC[N+](C)(C)CC1=CC=CC=C1 UREZNYTWGJKWBI-UHFFFAOYSA-M 0.000 description 1
- 235000019445 benzyl alcohol Nutrition 0.000 description 1
- 229960004217 benzyl alcohol Drugs 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- UCMIRNVEIXFBKS-UHFFFAOYSA-N beta-alanine Chemical group NCCC(O)=O UCMIRNVEIXFBKS-UHFFFAOYSA-N 0.000 description 1
- 229940000635 beta-alanine Drugs 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 239000007998 bicine buffer Substances 0.000 description 1
- 210000000941 bile Anatomy 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000001851 biosynthetic effect Effects 0.000 description 1
- 229930189065 blasticidin Natural products 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 229960003168 bronopol Drugs 0.000 description 1
- ITMIAZBRRZANGB-UHFFFAOYSA-N but-3-ene-1,2-diol Chemical compound OCC(O)C=C ITMIAZBRRZANGB-UHFFFAOYSA-N 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 159000000007 calcium salts Chemical class 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 150000001244 carboxylic acid anhydrides Chemical class 0.000 description 1
- 229920003123 carboxymethyl cellulose sodium Polymers 0.000 description 1
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 229940105329 carboxymethylcellulose Drugs 0.000 description 1
- 229940063834 carboxymethylcellulose sodium Drugs 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 229940106189 ceramide Drugs 0.000 description 1
- ZVEQCJWYRWKARO-UHFFFAOYSA-N ceramide Natural products CCCCCCCCCCCCCCC(O)C(=O)NC(CO)C(O)C=CCCC=C(C)CCCCCCCCC ZVEQCJWYRWKARO-UHFFFAOYSA-N 0.000 description 1
- 229960001927 cetylpyridinium chloride Drugs 0.000 description 1
- YMKDRGPMQRFJGP-UHFFFAOYSA-M cetylpyridinium chloride Chemical compound [Cl-].CCCCCCCCCCCCCCCC[N+]1=CC=CC=C1 YMKDRGPMQRFJGP-UHFFFAOYSA-M 0.000 description 1
- 229960003260 chlorhexidine Drugs 0.000 description 1
- KVSASDOGYIBWTA-UHFFFAOYSA-N chloro benzoate Chemical compound ClOC(=O)C1=CC=CC=C1 KVSASDOGYIBWTA-UHFFFAOYSA-N 0.000 description 1
- VIMWCINSBRXAQH-UHFFFAOYSA-M chloro-(2-hydroxy-5-nitrophenyl)mercury Chemical compound OC1=CC=C([N+]([O-])=O)C=C1[Hg]Cl VIMWCINSBRXAQH-UHFFFAOYSA-M 0.000 description 1
- FOCAUTSVDIKZOP-UHFFFAOYSA-N chloroacetic acid Chemical compound OC(=O)CCl FOCAUTSVDIKZOP-UHFFFAOYSA-N 0.000 description 1
- 229960002242 chlorocresol Drugs 0.000 description 1
- 229960003993 chlorphenesin Drugs 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000000975 co-precipitation Methods 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- GHVNFZFCNZKVNT-UHFFFAOYSA-M decanoate Chemical compound CCCCCCCCCC([O-])=O GHVNFZFCNZKVNT-UHFFFAOYSA-M 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 229950006137 dexfosfoserine Drugs 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- FFYPMLJYZAEMQB-UHFFFAOYSA-N diethyl pyrocarbonate Chemical compound CCOC(=O)OC(=O)OCC FFYPMLJYZAEMQB-UHFFFAOYSA-N 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-M dihydrogenphosphate Chemical compound OP(O)([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-M 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- XPPKVPWEQAFLFU-UHFFFAOYSA-J diphosphate(4-) Chemical class [O-]P([O-])(=O)OP([O-])([O-])=O XPPKVPWEQAFLFU-UHFFFAOYSA-J 0.000 description 1
- FRKBLBQTSTUKOV-UHFFFAOYSA-N diphosphatidyl glycerol Natural products OP(O)(=O)OCC(OP(O)(O)=O)COP(O)(O)=O FRKBLBQTSTUKOV-UHFFFAOYSA-N 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 235000019800 disodium phosphate Nutrition 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 229940018600 docusate potassium Drugs 0.000 description 1
- QBHFVMDLPTZDOI-UHFFFAOYSA-N dodecylphosphocholine Chemical compound CCCCCCCCCCCCOP([O-])(=O)OCC[N+](C)(C)C QBHFVMDLPTZDOI-UHFFFAOYSA-N 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 229950011333 edamine Drugs 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 229960004756 ethanol Drugs 0.000 description 1
- YMFBFFPJRABBPE-BTVCFUMJSA-N ethanol;(2r,3s,4r,5r)-2,3,4,5,6-pentahydroxyhexanal Chemical compound CCO.OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O YMFBFFPJRABBPE-BTVCFUMJSA-N 0.000 description 1
- 125000001301 ethoxy group Chemical group [H]C([H])([H])C([H])([H])O* 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 150000002187 fatty acyl carnitines Chemical class 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 239000001530 fumaric acid Substances 0.000 description 1
- 229960004675 fusidic acid Drugs 0.000 description 1
- IECPWNUMDGFDKC-MZJAQBGESA-N fusidic acid Chemical class O[C@@H]([C@@H]12)C[C@H]3\C(=C(/CCC=C(C)C)C(O)=O)[C@@H](OC(C)=O)C[C@]3(C)[C@@]2(C)CC[C@@H]2[C@]1(C)CC[C@@H](O)[C@H]2C IECPWNUMDGFDKC-MZJAQBGESA-N 0.000 description 1
- 150000008195 galaktosides Chemical class 0.000 description 1
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 description 1
- 230000030136 gastric emptying Effects 0.000 description 1
- 239000003629 gastrointestinal hormone Substances 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 229940093181 glucose injection Drugs 0.000 description 1
- 208000018914 glucose metabolism disease Diseases 0.000 description 1
- 229930195712 glutamate Natural products 0.000 description 1
- 229940049906 glutamate Drugs 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-M glutaminate Chemical compound [O-]C(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-M 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 108010049041 glutamylalanine Proteins 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000002641 glycemic effect Effects 0.000 description 1
- 229960002449 glycine Drugs 0.000 description 1
- KWIUHFFTVRNATP-UHFFFAOYSA-N glycine betaine Chemical compound C[N+](C)(C)CC([O-])=O KWIUHFFTVRNATP-UHFFFAOYSA-N 0.000 description 1
- 125000003630 glycyl group Chemical group [H]N([H])C([H])([H])C(*)=O 0.000 description 1
- 229940043257 glycylglycine Drugs 0.000 description 1
- 108010077515 glycylproline Proteins 0.000 description 1
- ZRALSGWEFCBTJO-UHFFFAOYSA-N guanidine group Chemical group NC(=N)N ZRALSGWEFCBTJO-UHFFFAOYSA-N 0.000 description 1
- 150000002367 halogens Chemical group 0.000 description 1
- 108010036302 hemoglobin AS Proteins 0.000 description 1
- MNWFXJYAOYHMED-UHFFFAOYSA-M heptanoate Chemical compound CCCCCCC([O-])=O MNWFXJYAOYHMED-UHFFFAOYSA-M 0.000 description 1
- KUIJPSLAGAQZTC-UHFFFAOYSA-N hexadecanoyl phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OP(O)(O)=O KUIJPSLAGAQZTC-UHFFFAOYSA-N 0.000 description 1
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 description 1
- 229920006158 high molecular weight polymer Polymers 0.000 description 1
- 108010045383 histidyl-glycyl-glutamic acid Proteins 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 102000053680 human centrocyte centroblast marker 1 Human genes 0.000 description 1
- 108700042450 human centrocyte centroblast marker 1 Proteins 0.000 description 1
- 238000006197 hydroboration reaction Methods 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- 239000000017 hydrogel Substances 0.000 description 1
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-M hydroxide Chemical compound [OH-] XLYOFNOQVPJJNP-UHFFFAOYSA-M 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 229940089468 hydroxyethylpiperazine ethane sulfonic acid Drugs 0.000 description 1
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 1
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Substances C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 1
- ATYGMDZIBQHJFX-UHFFFAOYSA-N imidazolidine;urea Chemical compound NC(N)=O.C1CNCN1 ATYGMDZIBQHJFX-UHFFFAOYSA-N 0.000 description 1
- 150000002461 imidazolidines Chemical class 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 230000003914 insulin secretion Effects 0.000 description 1
- 230000002473 insulinotropic effect Effects 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- CBEQRNSPHCCXSH-UHFFFAOYSA-N iodine monobromide Chemical compound IBr CBEQRNSPHCCXSH-UHFFFAOYSA-N 0.000 description 1
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
- 201000002529 islet cell tumor Diseases 0.000 description 1
- 210000004153 islets of langerhan Anatomy 0.000 description 1
- KQNPFQTWMSNSAP-UHFFFAOYSA-N isobutyric acid Chemical compound CC(C)C(O)=O KQNPFQTWMSNSAP-UHFFFAOYSA-N 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- FZWBNHMXJMCXLU-BLAUPYHCSA-N isomaltotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)O1 FZWBNHMXJMCXLU-BLAUPYHCSA-N 0.000 description 1
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 1
- JJWLVOIRVHMVIS-UHFFFAOYSA-N isopropylamine Chemical compound CC(C)N JJWLVOIRVHMVIS-UHFFFAOYSA-N 0.000 description 1
- 239000007951 isotonicity adjuster Substances 0.000 description 1
- 210000001630 jejunum Anatomy 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000003292 kidney cell Anatomy 0.000 description 1
- 201000006370 kidney failure Diseases 0.000 description 1
- 101150000548 laf1 gene Proteins 0.000 description 1
- 125000000400 lauroyl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000011866 long-term treatment Methods 0.000 description 1
- 125000003588 lysine group Chemical group [H]N([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 1
- 150000002672 m-cresols Chemical class 0.000 description 1
- 239000001095 magnesium carbonate Substances 0.000 description 1
- ZLNQQNXFFQJAID-UHFFFAOYSA-L magnesium carbonate Chemical compound [Mg+2].[O-]C([O-])=O ZLNQQNXFFQJAID-UHFFFAOYSA-L 0.000 description 1
- 235000014380 magnesium carbonate Nutrition 0.000 description 1
- 229960001708 magnesium carbonate Drugs 0.000 description 1
- 229910000021 magnesium carbonate Inorganic materials 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 230000005389 magnetism Effects 0.000 description 1
- IWYDHOAUDWTVEP-UHFFFAOYSA-N mandelic acid Chemical compound OC(=O)C(O)C1=CC=CC=C1 IWYDHOAUDWTVEP-UHFFFAOYSA-N 0.000 description 1
- 229960002510 mandelic acid Drugs 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 125000005341 metaphosphate group Chemical group 0.000 description 1
- 150000005342 methoxybenzoic acids Chemical class 0.000 description 1
- RMAHPRNLQIRHIJ-UHFFFAOYSA-N methyl carbamimidate Chemical compound COC(N)=N RMAHPRNLQIRHIJ-UHFFFAOYSA-N 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- NEGQCMNHXHSFGU-UHFFFAOYSA-N methyl pyridine-2-carboximidate Chemical compound COC(=N)C1=CC=CC=N1 NEGQCMNHXHSFGU-UHFFFAOYSA-N 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000003607 modifier Substances 0.000 description 1
- NJTGANWAUPEOAX-UHFFFAOYSA-N molport-023-220-454 Chemical compound OCC(O)CO.OCC(O)CO NJTGANWAUPEOAX-UHFFFAOYSA-N 0.000 description 1
- 229940045641 monobasic sodium phosphate Drugs 0.000 description 1
- VYQNWZOUAUKGHI-UHFFFAOYSA-N monobenzone Chemical compound C1=CC(O)=CC=C1OCC1=CC=CC=C1 VYQNWZOUAUKGHI-UHFFFAOYSA-N 0.000 description 1
- PJUIMOJAAPLTRJ-UHFFFAOYSA-N monothioglycerol Chemical compound OCC(O)CS PJUIMOJAAPLTRJ-UHFFFAOYSA-N 0.000 description 1
- 201000006417 multiple sclerosis Diseases 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- VVGIYYKRAMHVLU-UHFFFAOYSA-N newbouldiamide Natural products CCCCCCCCCCCCCCCCCCCC(O)C(O)C(O)C(CO)NC(=O)CCCCCCCCCCCCCCCCC VVGIYYKRAMHVLU-UHFFFAOYSA-N 0.000 description 1
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 1
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 150000002883 o-cresols Chemical class 0.000 description 1
- AQFWNELGMODZGC-UHFFFAOYSA-N o-ethylhydroxylamine Chemical compound CCON AQFWNELGMODZGC-UHFFFAOYSA-N 0.000 description 1
- 235000020824 obesity Nutrition 0.000 description 1
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 1
- 239000006186 oral dosage form Substances 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 150000007530 organic bases Chemical class 0.000 description 1
- 229960003104 ornithine Drugs 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 150000002931 p-cresols Chemical class 0.000 description 1
- 239000006174 pH buffer Substances 0.000 description 1
- 125000001312 palmitoyl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 208000022102 pancreatic neuroendocrine neoplasm Diseases 0.000 description 1
- 206010033675 panniculitis Diseases 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000032696 parturition Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 230000006320 pegylation Effects 0.000 description 1
- JLFNLZLINWHATN-UHFFFAOYSA-N pentaethylene glycol Chemical compound OCCOCCOCCOCCOCCO JLFNLZLINWHATN-UHFFFAOYSA-N 0.000 description 1
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 description 1
- 150000008104 phosphatidylethanolamines Chemical class 0.000 description 1
- 150000003905 phosphatidylinositols Chemical class 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- HMFAQQIORZDPJG-UHFFFAOYSA-N phosphono 2-chloroacetate Chemical class OP(O)(=O)OC(=O)CCl HMFAQQIORZDPJG-UHFFFAOYSA-N 0.000 description 1
- 150000003016 phosphoric acids Chemical class 0.000 description 1
- BZQFBWGGLXLEPQ-REOHCLBHSA-N phosphoserine Chemical compound OC(=O)[C@@H](N)COP(O)(O)=O BZQFBWGGLXLEPQ-REOHCLBHSA-N 0.000 description 1
- XNGIFLGASWRNHJ-UHFFFAOYSA-L phthalate(2-) Chemical compound [O-]C(=O)C1=CC=CC=C1C([O-])=O XNGIFLGASWRNHJ-UHFFFAOYSA-L 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 150000003053 piperidines Chemical class 0.000 description 1
- 230000001817 pituitary effect Effects 0.000 description 1
- 229920001993 poloxamer 188 Polymers 0.000 description 1
- 229940044519 poloxamer 188 Drugs 0.000 description 1
- 229920001992 poloxamer 407 Polymers 0.000 description 1
- 229940044476 poloxamer 407 Drugs 0.000 description 1
- 229920000747 poly(lactic acid) Polymers 0.000 description 1
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 229920002635 polyurethane Polymers 0.000 description 1
- 239000004814 polyurethane Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- UORVCLMRJXCDCP-UHFFFAOYSA-M propynoate Chemical compound [O-]C(=O)C#C UORVCLMRJXCDCP-UHFFFAOYSA-M 0.000 description 1
- 229940048914 protamine Drugs 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 108060006633 protein kinase Proteins 0.000 description 1
- 230000002797 proteolythic effect Effects 0.000 description 1
- 230000006337 proteolytic cleavage Effects 0.000 description 1
- 235000019423 pullulan Nutrition 0.000 description 1
- 229960003581 pyridoxal Drugs 0.000 description 1
- 235000008164 pyridoxal Nutrition 0.000 description 1
- 239000011674 pyridoxal Substances 0.000 description 1
- 235000007682 pyridoxal 5'-phosphate Nutrition 0.000 description 1
- 239000011589 pyridoxal 5'-phosphate Substances 0.000 description 1
- 229960001327 pyridoxal phosphate Drugs 0.000 description 1
- 125000001453 quaternary ammonium group Chemical group 0.000 description 1
- 238000007348 radical reaction Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000012340 reverse transcriptase PCR Methods 0.000 description 1
- 238000009666 routine test Methods 0.000 description 1
- YGSDEFSMJLZEOE-UHFFFAOYSA-M salicylate Chemical compound OC1=CC=CC=C1C([O-])=O YGSDEFSMJLZEOE-UHFFFAOYSA-M 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 229940116351 sebacate Drugs 0.000 description 1
- CXMXRPHRNRROMY-UHFFFAOYSA-L sebacate(2-) Chemical compound [O-]C(=O)CCCCCCCCC([O-])=O CXMXRPHRNRROMY-UHFFFAOYSA-L 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000007493 shaping process Methods 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 210000003625 skull Anatomy 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 235000017550 sodium carbonate Nutrition 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- NRHMKIHPTBHXPF-TUJRSCDTSA-M sodium cholate Chemical compound [Na+].C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC([O-])=O)C)[C@@]2(C)[C@@H](O)C1 NRHMKIHPTBHXPF-TUJRSCDTSA-M 0.000 description 1
- 229940079839 sodium dehydroacetate Drugs 0.000 description 1
- 235000019259 sodium dehydroacetate Nutrition 0.000 description 1
- FHHPUSMSKHSNKW-SMOYURAASA-M sodium deoxycholate Chemical compound [Na+].C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC([O-])=O)C)[C@@]2(C)[C@@H](O)C1 FHHPUSMSKHSNKW-SMOYURAASA-M 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- IHQKEDIOMGYHEB-UHFFFAOYSA-M sodium dimethylarsinate Chemical compound [Na+].C[As](C)([O-])=O IHQKEDIOMGYHEB-UHFFFAOYSA-M 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- OABYVIYXWMZFFJ-ZUHYDKSRSA-M sodium glycocholate Chemical compound [Na+].C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCC([O-])=O)C)[C@@]2(C)[C@@H](O)C1 OABYVIYXWMZFFJ-ZUHYDKSRSA-M 0.000 description 1
- BYKRNSHANADUFY-UHFFFAOYSA-M sodium octanoate Chemical compound [Na+].CCCCCCCC([O-])=O BYKRNSHANADUFY-UHFFFAOYSA-M 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 235000011008 sodium phosphates Nutrition 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- JAJWGJBVLPIOOH-IZYKLYLVSA-M sodium taurocholate Chemical compound [Na+].C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCCS([O-])(=O)=O)C)[C@@]2(C)[C@@H](O)C1 JAJWGJBVLPIOOH-IZYKLYLVSA-M 0.000 description 1
- DSOWAKKSGYUMTF-GZOLSCHFSA-M sodium;(1e)-1-(6-methyl-2,4-dioxopyran-3-ylidene)ethanolate Chemical compound [Na+].C\C([O-])=C1/C(=O)OC(C)=CC1=O DSOWAKKSGYUMTF-GZOLSCHFSA-M 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 150000003408 sphingolipids Chemical class 0.000 description 1
- WWUZIQQURGPMPG-KRWOKUGFSA-N sphingosine Chemical compound CCCCCCCCCCCCC\C=C\[C@@H](O)[C@@H](N)CO WWUZIQQURGPMPG-KRWOKUGFSA-N 0.000 description 1
- 230000003019 stabilising effect Effects 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 230000003335 steric effect Effects 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- TYFQFVWCELRYAO-UHFFFAOYSA-L suberate(2-) Chemical compound [O-]C(=O)CCCCCCC([O-])=O TYFQFVWCELRYAO-UHFFFAOYSA-L 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-L sulfite Chemical compound [O-]S([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-L 0.000 description 1
- 150000003467 sulfuric acid derivatives Chemical class 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 238000011287 therapeutic dose Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 239000008181 tonicity modifier Substances 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 239000012096 transfection reagent Substances 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000010474 transient expression Effects 0.000 description 1
- 238000003146 transient transfection Methods 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- 125000001493 tyrosinyl group Chemical group [H]OC1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 230000037303 wrinkles Effects 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
- 229960003487 xylose Drugs 0.000 description 1
- 150000003751 zinc Chemical class 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/605—Glucagons
-
- 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
- A61P25/28—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
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/04—Anorexiants; Antiobesity agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
- A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pharmacology & Pharmacy (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Diabetes (AREA)
- Endocrinology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Toxicology (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Biomedical Technology (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Obesity (AREA)
- Hematology (AREA)
- Hospice & Palliative Care (AREA)
- Child & Adolescent Psychology (AREA)
- Psychiatry (AREA)
- Emergency Medicine (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
本发明提供了具有GLP-1活性和增加的在体稳定性的融合肽,该融合肽特别地对IV型糖尿病具有抗性。该融合肽包括作为组分(I)的N-端GLP-1(7-35、7-36或7-37)序列,和作为组分(II)的C-端具有至少9个氨基酸的肽序列或其功能型片段、变异体或衍生物。组分(II)优选为IP2(插入肽2)的完整或部分版本。优选的实施例包括序列GLP-1(7-35、36或37)/IP2/GLP-1(7-35、36或37)或GLP-2。该融合肽可在工程细胞中生成或合成,且可用于制备用于治疗各种疾病或失调的药物,例如1或2型糖尿病、下垂症相关疾病或神经退行性失调等。
Description
技术领域
本发明涉及新型胰高血糖素样融合肽-1(GLP-1 fusion peptides),其具有延伸的C-端,并耐内肽酶IV失活,可在动物转化细胞中以较高水平表达,且可用于例如2型糖尿病的治疗。
背景技术
胰高血糖素基因已经得到广泛的研究(参见White,J.W.et al.,1986 NucleicAcid Res.14(12)4719-4730)。作为高分子量前体分子,胰高血糖素前原分子在胰腺α细胞、空肠和结肠L细胞中合成。胰高血糖素前原是包括180个氨基酸的长激素原,且除了胰高血糖素外其序列还包括两个结构相关的序列:胰高血糖素样肽-1(GLP-1)和胰高血糖素样肽-2(GLP-2)。在胰高血糖素前原分子中,GLP-1和GLP-2之间是插入肽2(IP2)的17肽氨基酸序列(或可以是15肽氨基酸序列加C-端RR裂解位点)。IP2序列(位于前元分子的GLP-1和GLP-2之间)通常在GLP-1的aa 37后通过蛋白水解裂解。因此,胰高血糖素前原分子根据细胞和环境裂解成不同的肽,包括GLP-1(1-37)、和未加工形态的37肽氨基酸。通常,该加工过程通常发生在胰腺和肠中。可将该GLP-1(1-37)序列进一步蛋白水解加工以获得活性GLP-1(7-37)、31肽氨基酸加工形态、或GLP-1(7-36)醯胺。因此,命名GLP-1(7-36)指出了我们所讨论的氨基酸片段包括从亲本肽(parent peptide)GLP-1的N-端起的第7号(包含7号)到第37号(包含37号)的氨基酸残基。分子式1(SEQ ID No.:43)示出了GLP-1(7-36)醯胺和GLP-1(7-37)的氨基酸序列:
His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-X(I),当X是NH2时,该分子式示出的是GLP-1(7-36)醯胺,当X是Gly-OH时,示出的是GLP-1(7-37)。
GLP-1是胃肠道激素,其是最有效的内源性促胰岛素剂,并能在β细胞中提高腺苷酸环化酶和蛋白激酶活性。生理学上,GLP-1和来自上部肠道的肠抑胃肽共同用作肠促胰岛激素,来降低血糖水平。因此,例如响应食物摄取而分泌的GLP-1在胃、肝脏和脑部具有多方面的效应,协同工作以调节血糖。由于胰高血糖素样肽-1 GLP-1(7-36)醯胺和其非酰胺化类似物GLP-1(7-37)在碳水化合物代谢上的有效作用和其潜在的疾病治疗应用(包括2型糖尿病),因此具有相当大的吸引力。2型糖尿病的特征在于抗胰岛素性,因此当胰岛素存在时,细胞并不对其做出合适的响应。其比1型糖尿病更为复杂。由于其症状一般较轻微(没有酮酸中毒)并是偶发性的,因此在被诊断出来以前,2型糖尿病可能在病人体内潜伏很久。然而,被忽视的2型糖尿病可能导致严重的并发症,包括肾衰竭和冠心病。2型糖尿病将导致这些疾病的发病率和死亡率的增加。
GLP-1(7-36)醯胺或GLP-1(7-37)在血液中存活的时间很短。二肽酰肽酶IV(DPP-IV)将肽从残基8和9之间裂解。裂解后的肽无活性。由于外源GLP-1的存活时间相当地短,因此限制了其在治疗上的应用。
为了合成天然生成的GLP-1(GLP-1(7-37))的稳定(对抗DPP-IV)类似物,已经做出了各种尝试。特别地,在活体内为丙氨酸(Ala)的残基8被替换成如甘氨酸(Gly)、丝氨酸(Ser)或苏氨酸(Thr)(参见Burcelin,R.,et al.(1999)Metabolism 48,252-258)。对Gly8或G8进行广泛地测试,它们均是合成分子,并由细胞株基因工程生成以分泌突变多肽(参见Burcelin,R.,et al(1999)Annals of the New York Academy of Sciences 875:277-285)。可将各种其它的修饰引入GLP-1(7-37)以加强其在体稳定性且并不影响其生物活性。然而,由于牵涉到相当多的难题,所有的这些方法都难以取得显著的治疗效果。
在WO/9953064中,Thorens,B.公开了一种用于制作多聚体GLP-1表达盒的方法,其可与多种细胞类型合并,这些细胞是广泛应用的永存细胞株并采用不同的原始细胞培养。样本可包括来自哺乳动物的CNS的响应EGF的神经球(neurospheres)、响应bFGF的神经元干细胞,处理过的样本使用幼地鼠肾(BHK)细胞。据说,该移植转染细胞已经成功地用于身染糖尿病的老鼠,其可使血糖调节基本等于非糖尿病调节。然而,这个技术不符合可应用于糖尿病人的治疗的日常管理要求。
另一种外源稳定血糖水平的方法基于一类已知的新药,肠促胰岛素模拟剂(incretin mimetics),其对2型糖尿病的治疗正在研究中。依克那肽()是从毒蜥蜴(Gila monster lizard)唾液中获取的天然化合物的合成版本。在临床试验中,已经证实肠促胰岛素模拟剂(依克那肽)可降低血糖并改进β细胞功能。然而,依克那肽仅仅显示了人类的肠促胰岛素胰高血糖素样肽-1(GLP-1)的某些功效。
总之,对于2型糖尿病,现在还没有有效的治疗方法可以允许在GLP-1的基础上降低血糖水平,换句话说,还不能提供一种可反应GLP-1已知的所有有利作用的疗法。举例来说,通过降低肥胖患者的胃排空率或胰岛素刺激活性,具有生理浓度活性的GLP-1有效地降低了进入血液的营养物质的比率。
发明内容
因此,本发明的目的是提供基于GLP-1的肽分子,其具有生物活性并能抗蛋白降解。
本发明涉及一种融合肽,包括作为组分(I)N-端的GLP-1(7-35,7-36或7-37)序列和作为组分(II)C-端的包括至少9个氨基酸的肽序列或其功能性片段、变异体或衍生物。
本发明基于以下发现:生成的本发明肽在活体内能抵抗蛋白降解,这主要是取决于蛋白水解内肽酶(proteolytic endopeptidase)IV的活性。本发明具有至少两个组分(I)和(II)的肽,呈现出GLP-1的生物活性,同时通过对其组分(I)进行C-端延长能赋予GLP-1稳定性。
此处所用的术语“本发明肽”是在此定义的融合肽以及其变异体、类似物、片段或衍生物,包括例如融合肽的衍生片段、类似物或变异体的结合。此处所用的术语“GLP-1肽”表示GLP-1(7-35、36或37),而“经修饰的GLP-1肽”表示任何GLP-1类似物、GLP-1衍生物、GLP-1变异体或GLP-1片段,包括GLP-1(7-35、36或37)的衍生片段、类似物或变异体,这些可存在于本发明肽的组分(I)和(II)中。此处所用的术语“GLP-2肽”表示GLP-2(1-33、34、或35),而“经修饰的GLP-2肽”表示任何GLP-2类似物、片段或变异体、GLP-2衍生物或GLP-2类似物的衍生物,包括GLP-2(1-33、34、或35)的衍生片段、类似物或变异体。可将这些变异体、类似物、片段和衍生物分类成未修饰序列(举例来说,GLP-1(7-35、36或37)或GLP-2(1-33、34或35))的修饰物。在本发明中,任何变异体、类似物、片段或衍生物都是功能性的,举例来说,能够完成和未修饰的GLP-1肽相同或相类似的功能。
优选地,本发明肽是融合肽或其变异体、类似物、片段或衍生物,其中,组分(I)包含一个序列,其至少与SEQ ID No.:1具有80%的序列同系性,更佳地与SEQ ID No.:1具有85%的序列同系性,最好与SEQ ID No.:1具有90%的序列同系性。SEQ ID No.:1呈现GLP-1(7-37)(长度为31个氨基酸)的天然氨基酸序列,该序列在哺乳动物间严格保持不变。
根据本发明的融合肽(或更普遍地包括该融合肽的类似物、片段、变异体或衍生物的任何本发明肽)的第二组分(组分(II))一般包含长度为至少9个氨基酸的序列,该序列可以构成β-回转状结构,也可以不构成β-回转状结构。β-回转状结构是蛋白质或肽的典型二级结构元,其一般由4个氨基酸构成,并从肽或是蛋白质的主链方向反转。组分(II)的氨基酸序列包括至少9个氨基酸,更佳地在其序列中包含至少一个脯氨酸或丙胺酸残基。脯氨酸残基是β-回转构象四聚体氨基酸序列中的常见氨基酸。脯氨酸残基一般位于融合肽的组分(II)中存在的四聚物β-回转序列的位2或3,优选位2。
本发明融合肽的组分(II)序列一般长度为9到30个氨基酸,更佳地长度为9到20个氨基酸,最好长度为9到15个氨基酸。一般而言,由于与较长序列相比,组分(II)中的较短序列具有更高的与GLP受体结合活性,因而优选较短序列。虽然不是先觉条件,组分(II)的序列可优选中性的或可在pH7带负电荷。
根据本发明的所述的融合肽,优选其组分(II)包含从VAIA、IAEE、PEEV、AEEV、EELG、AAAA、AAVA、AALG、DFPE、AADX、AXDX和XADX中选出的序列基元,其中X表示任何氨基酸(天然氨基酸或修饰后的非天然氨基酸)。这些四聚体基元可位于组分(II)序列的任何位置。在特定的优选实施例中,本发明融合肽的组分(II)是通过其N-端序列基元与组分(I)的C-端链接的肽序列,其N-端序列基元选自AA、XA、AX、RR、RX和XR中,其中X表示任何氨基酸(天然氨基酸或修饰后的非天然氨基酸)。
本发明融合肽中的组分(II)的较优基元包含序列基元SEQ ID No.:25(DFPEEVA)或包含至少与序列基元DFPEEVA具有80%的序列同系性的序列。该序列基元DFPEEVA对应于人类或小鼠IP-2的部分序列。
在特别优选的融合肽中,组分(II)是肽序列,其包含依照SEQ ID No.:22(RRDFPEEVAI)或SEQ ID No.:26(AADFPEEVAI)的序列(所有的肽序列服从该单字母代码)或包含一个至少与SEQ ID No.:22或SEQ ID No.:26具有80%的序列同系性的序列。SEQ ID No.:22是全长(人类或小鼠)IP-2(插入肽)序列的部分序列,其包含15个氨基酸长度的全长IP-2序列中的N-端的10个氨基酸。SEQ ID No.:26是通过用于(AA)替换SEQ ID No.:22的N-端(RR)残基衍生出来的。IP-2是含氨基酸序列的β-回转的优选实施例。因此,组分(II)中包括的其它更优选的序列是IP-2的较长的部分氨基酸序列,例如人类中存在的N-端14氨基酸序列(SEQ ID No.:23(RRDFPEEVAIVEEL))or其小鼠配对物(SEQ ID No.24(RRDFPEEVAIAEEL))或至少与SEQ ID Nos.:23或24具有80%序列同系性的序列。融合肽的组分(II)中所含的最优成分是天然存在的IP-2序列的所有15个氨基酸的全长IP-2序列(SEQ ID No.:2(RRDFPEEVAIVEELG)人类或SEQ ID No.3(RRDFPEEVAIAEELG)小鼠)或至少与SEQ ID Nos.:2或3具有80%的序列同系性的序列。本发明的保护范围同样包括IP2的所有母乳动物异构体(母乳动物中的IP2的天然变异体)。本段中提到的所有序列也可具有N-端基元(AA)、(AX)或(XA)以替代天然基元(RR),举例来说,SEQ ID No.:27(AADFPEEVAIVEEL)、SEQ ID No.:28(AADFPEEVAIAEEL)、SEQ ID No.:29(AADFPEEVAIVEELG)、和SEQ IDNO.:30(AADFPEEVAIAEELG)。可多次复制组分(II)中包含的序列,举例来说,对IP2或IP2的片段、变异体、类似物或衍生物进行2次、3次或更多次复制。
因此,发明肽优选包含依照SEQ ID No.:8(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFPEEVAIAEELG)的序列,也就是,GLP-1(7-37)通过其C-端与小鼠IP2链接而不经任何接头序列(linker sequence ),或依照SEQ ID No.:12(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFPEEVAIVEELG)的序列,也就是,GLP-1(7-37)通过其C-端与人类IP2链接而不经任何接头序列。SEQ ID No.:8和SEQ ID No.:9的本发明肽的更佳的变异体是:
SEQ ID No.:31
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGAADFPEEVAIAEELG);
SEQ ID No.:32
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFAEEVAIAEELG);
SEQ ID No.:33
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDAAAAVAIAEELG);
SEQ ID No.:34
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGAADAAAAVAIAAALG);
SEQ ID No.:35
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFP);
SEQ ID No.:36
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFPEEVA);
SEQ ID No.:37
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFPEEVAIAEELGRRHAC);
SEQ ID No.:38
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGAADFPEEVAIVEELG);
SEQ ID No.:39
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFAEEVAIVEELG);
SEQ ID No.:40
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDAAAAVAIVEELG);
SEQ ID No.:41
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGAADAAAAVAIVAALG);
SEQ ID No.:42
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFPEEVAIVEELGRRHAC);
也就是,GLP-1(7-37)通过其C-端与IP2序列的特殊类似物或变异体链接而不经任何接头序列。优选还包括其变异体、类似物或片段,与SEQ ID Nos:8和12及它们的衍生物具有至少80%序列同系性。
在不违背任何理论的情况下,本发明的发明人得出如下结论:如果将其运用于任何危急情况下的病人,GLP-1(7-35、36或37)的不稳定性取决于其无保护的三维结构。蛋白酶可裂解GLP-1(7-35,36或37)肽并迅速废止其在体的生理活性。通过将肽序列链接到GLP-1(7-35,36or 37)的C-端,其结构将获得针对酶降解的稳定性。如果由于由其一级结构形成的β-回转结构成分的出现并为组分(II)提供硬度,延长的C-端肽序列(包含在根据本发明所述的融合肽的组分(II)中)折回,将增强获得的针对酶降解的稳定性。然而,本发明肽的组分(II)中的β-回转结构并不是组分(I)的GLP-1获得针对酶降解的稳定性的先决条件。由于其C-端肽延伸,例如包含β-回转结构成分,发现本发明肽改善了抗DPP-IV失活性质。在对目标细胞中的受体起作用之前,C-端肽并不从GLP-1(7-35、36或37)序列上裂解,或者可对C-端肽进行酶裂解以形成在体GLP-1(7-35、36或37)。不管本发明肽在GLP-1受体处的精确形式的如何,本发明肽作为活性促胰岛素化合物发挥其作用。
由于其构成β-回转成份的一级结构,适合包含在组分(II)中的肽序列很容易被正确识别,例如通过光谱法、图二色谱、或技术人员已知的其它方法)。
组分(II)和组分(I)可直接链接或通过接头序列来链接。两个组分优选彼此直接连接。在它们是通过接头序列连接的例子中,该接头序列优选肽接头或有机接头。肽接头的长度一般是1到10个氨基酸,更佳的肽接头长度是1到5个氨基酸,最优的肽接头长度是1到3个氨基酸,在某些例子中,接头序列可以更长,包括10到50个氨基酸。肽接头可由多种氨基酸序列组成。优选地,肽接头将在被连接的组分间引入某些结构柔性。结构柔性可通过例如包含各种甘氨酸或脯氨酸残基的肽接头来得到,优选在接头序列中的脯氨酸和甘氨酸残基至少占30%,至少占40%更佳,至少占60%为最佳。不管其特定序列如何,肽接头优选是免疫惰性的。
在本发明的优选实施例中,本发明肽,也就是融合肽或其类似物、片段、变异体或衍生物,包含第三组分(组分(III)),其与组分(II)的C-端连接和/或与组分(I)的N-端连接。优选地,组分(III)位于组分(II)的C-端。不管组分(III)是连接在组分(I)的N-端(通过其C-端),还是连接在组分(II)的C-端(通过其N-端),该连接可以是直接连接或是通过接头序列的间接连接。关于接头序列,其指的是前面公开的用于连接组分(I)和组分(II)的所有接头。通常,组分(III)包括至少4个氨基酸残基,更佳地包括至少10个附加氨基酸残基,最优地包括至少20或至少30个氨基酸残基。在功能方面中,组分(III)用于进一步增强本发明肽的稳定性。组分(III)将不会干扰本发明肽的生物学功能,本发明肽的生物活性明显可与GLP-1(7-37)的生物活性相比。
优选地,本发明肽的组分(III)包括至少4个附加的氨基酸残基,优选至少10个,更佳为至少20个,该氨基酸残基是任何哺乳动物机体(哺乳动物间的GLP-2的其它天然存在的变异体)的亚型GLP-2的N-端序列的氨基酸残基,例如SEQ ID Nos:4和5中示出的小鼠或人类亚基。GLP-2存在于胰高血糖素原,并在糖代谢中涉及。由于具有组分(I)(GLP-1肽)中包含的生物活性序列,组分(III)也可包括GLP-2的天然存在形式的类似物、变异体或衍生物。作为替换地另一种情况下,组分(III)也可包括至少4个GLP-1(7-37)的N-端序列的附加氨基酸残基,更佳地包括至少10个GLP-1(7-37)的N-端序列的附加氨基酸残基,最佳地包括至少20个GLP-1(7-37)的N-端序列的附加氨基酸残基,相应地包括所有哺乳动物亚型-如上所述-以及其功能性变异体、类似物或衍生物。一般而言,组分(III)可包括任何形式的GLP-1肽或修饰后的GLP-1肽,这些GLP-1肽也可适用于本发明肽的组分(I)。在进一步的可替换的实施例中,组分(III)也可包含GLP-1(7-37)和GLP-2的嵌合形式。该嵌合形式可通过将GLP-1(7-37)和GLP-2(或两者的片段、类似物、变异体或衍生物)彼此相连生成,随后将该嵌合形式作为组分(III)引入本发明肽。优选地,该嵌合形式由GLP-1(7-37)的部分序列和GLP-2的部分序列链接在一起组成。举例来说,该嵌合形式可包括GLP-1的N-端5到30个氨基酸和GLP-2的C-端5到30个氨基酸,或者反之亦然,例如,GLP-1(7-37)的氨基酸7或8到22、23、24、25、26、27或28和GLP-2的从15、16、17、18、19、20、21、22、23位到例如C-端的氨基酸序列。
如果将GLP-2或GLP-1(7-37)的天然存在形式的修饰分别用作组分(III),组分(III)优选分别包含SEQ ID Nos.:4或5或SEQ ID No.:1的序列,或包含有至少与SEQ ID Nos.:4或5或SEQ ID No.:1具有80%的序列同系性的序列。由于侧链修饰或肽主链修饰等(如在此公开的所属衍生物),本发明的组分(III)也可包含这些优选序列的衍生物。
在另一实施例中,组分(III)可包括多个如上所述的序列。举例来说,组分(III)可包括至少两个GLP-1(7-37)和/或GLP-2的拷贝,优选2、3或4个GLP-1(7-37)和/或GLP-2的拷贝,或包括与SEQ ID Nos:1、4或5具有80%序列同系性的序列的至少两个拷贝。组分(III)也可包括如上所述的更多的GLP-1(7-37)或GLP-2的嵌合版本的拷贝,最终形成嵌合版本和GLP-1(7-37)和/或GLP-2的结合物或其具有至少80%序列同源性的修饰版本。在本发明的保护范围中,包括两个或更多个组分(III),其中优选两个组分(III),举例来说:(1)可通过其N-端直接或间接连接到组分(II)的C-端且;(2)通过其C-端直接或间接连接到组分(I)的N-端。如果提供了两个组分(III),这两个组分(III)可以是相同的,也可以是不同的。
因此,本发明的融合肽特别优选包含三个组分(I)、(II)和(III)。包含所有这三个组分的四个实施例从以下组中选出:SEQ ID No.6(N-GLP-1(7-37)-IP2(小鼠)-RR-GLP-1(7-37)-C,在此又称为小鼠CM2)、SEQ IDNo.7(N-GLP-1(7-37)-IP2(小鼠)-RR-GLP2-C,在此又称为小鼠CM2)、SEQ IDNo.10(N-GLP-1(7-37)-IP2(人类)-RR-GLP-1(7-37)-C,在此又称为人类CM1)和SEQ ID No.11(N-GLP-1(7-37)-IP2(人类)-RR-GLP-2-C),在此又称为人类CM2),或者是与SEQ ID Nos.:6、7、10或11具有至少80%序列同系性的序列或其衍生物。所有序列6、7、10和11在IP2(组分(II))的C-端包含有RR-接头(两个精氨酸残基),该接头也可以被去除。在依照SEQ ID Nos:6、7、10或11的每个实施例中的组分(I)是GLP-1(7-37),而组分(III)(在每个实施例中其与组分(II)的C-端相连)是GLP-1(7-37)或GLP-2。
本发明肽可在以多种修饰形式存在。下面将对这些修饰形式进行详细描述。
本申请中,术语“盐”既指羧基盐,也指上述的融合肽的氨基酸加成盐或它们的类似物、片段、衍生物或变异体。羧基盐可通过本领域已知的方法制备,包括无机盐,如钠盐、钙盐、铵盐、铁盐或锌盐等等,以及有机碱盐,采用如胺、乙醇胺、精氨酸或赖氨酸、哌啶、普鲁卡因等等制备。酸加成盐包括:如无机酸盐,该无机酸可以如盐酸或硫酸,以及有机酸盐,该有机酸可以如乙酸或草酸。当然,所有这些盐都必须保持本发明肽的生物活性,也就是,降低营养物质进入血液循环的速率的能力。如下所述,肽盐(salt peptide)形式将包含在制剂分子式中。
根据本发明的融合肽的“片段”是指该分子的任意子集,即可保留期望的生物活性的较短肽。通过在该分子的任意端切除氨基酸并检验其是否具有肠胰岛素的性质,就可容易地制备所述片段。用于每次从多肽的N-端和/或C-端切除一个氨基酸的蛋白酶是已知的,因此确定保持期望的生物活性仅仅是例行试验。最后,片段可取决于肽端的氨基酸的切除和/或所切除的氨基酸在肽序列中的位置。
另外,本发明肽具有抗2型糖尿病活性,其本身作为融合肽及其类似物或变异体、盐、功能性衍生物和/或片段,可还包含有侧接在本发明肽两端的附加氨基酸残基。只要该合成分子保持其对蛋白酶的抗性和稳定性,以及其作为肠促胰岛素的性能,就能确定是否有任何这样的侧接残基影响中心肽的基本和新颖特性,举例来说,通过其对胰腺细胞的作用,通过例行实验。当涉及某个特定序列时,用语“本质上由……组成”是可以存在一些附加的侧接残基而不会影响本发明肽的基本和新颖特性。这一用语并不包括该特定序列中的取代、切除或添加。
根据本发明所述的“变异体”指与以上定义的整个肽或其片段基本类似的分子。使用本领域已知的方法,可以通过直接化学合成便利地制备变异肽(variant peptides)。当然,本发明肽的这些变异体将具有类似的抗糖尿病功效,举例来说,与相应的天然存在的GLP-1肽相同的胰岛素刺激活性。
可替换地另一种情况下,以上定义的肽的氨基酸序列变异体可由编码合成衍生物的DNA的突变来制备。这些变异体包括例如从氨基酸序列中切除残基、或在氨基酸序列中插入残基或取代氨基酸序列中的残基。假定最后的产物具有期望的活性,可通过切除、插入、取代的任意组合来获得最后产物。显然地,在对编码该变异肽的DNA做出的突变必须不会改变阅读框,并且更佳地不会形成能生成二级mRNA结构的互补区域。
根据本发明以上定义的肽的“类似物”指与整个分子或其活性片段基本类似的非天然分子。这些类似物将展现与对应的天然存在的GLP-1肽相同的活性。
可在根据本发明所述的肽中做出的置换类型可取决于对在不同种类的同源蛋白/肽之间氨基酸变化的频率的分析。基于这些分析,在此可将存性置换(conservative substitution)定义为在下列五个基团中的一个中的调换:
I小的、脂肪族、非极性的或弱极性的残基:Ala(丙氨酸)、Ser(丝氨酸)、Thr(苏氨酸)、Pro(脯氨酸)、Gly(甘氨酸);
II.极性的、带负电荷的残基和它们的醯胺:Asp(门冬氨酸)、Asn(天冬酰胺)、Glu(谷氨酸)、Gln(谷氨酰胺);
III.极性的、带正电荷的残基:His(组氨酸)、Arg(精氨酸)、Lys(赖氨酸);
IV.大的、芬芳族、非极性的残基:Phe(苯丙氨酸)、Tyr(酪氨酸)、Trp(色氨酸)。
在前述基团中,下列置换可看作是“高保存性的”:Asp/Glu;His/Arg/Lys;Phe/Tyr/Trp;Met/Leu/Ile/Val。半保存性置换定义为以上基团(I)-(IV)中的两个之间的替换,限于大基团(supergroup)(A),包括上述(I)、(II)和(III),或限于大基团(B),包括上述(IV)和(V)。置换并不限于基因编码以至天然存在的氨基酸。
通常,本发明肽的类似物或变异体也可包含有氨基酸置换,举例来说,做出此类置换具有改进其溶解性的目的(用亲水性氨基酸取代疏水性氨基酸)。在本发明肽的GLP-1肽的变异体/类似物(存在于本发明肽的组分(I)和/或(III))的实施例中,(经修饰的)GLP-1肽的特征在于在GLP-1肽的7、8、11、12、16、22、23、23、25、27、30、33、34、35、36或37位的一个或多个置换。作为下列命名的一个例子,[Arg34-GLP-1(7-37)]指的是GLP-1类似物,其中位于34位的天然存在的赖氨酸被精氨酸取代。
特别地,本发明肽的组分(I)和/或组分(III)可包括GLP-1(7-35、36或37)的变异体和类似物,包括例如Gln9-GLP-1(7-37)、D-Gln9-GLP-1(7-37)、acetyl-Lys9-GLP-1(7-37)、Thr16-Lys18-GLP-1(7-37)和Lys18-GLP-1(7-37)、Arg34-GLP-1(7-37)、Lys38-Arg26-GLP-1(7-38)-OH、Lys36-Arg26-GLP-1(7-36)、Arg26,34-Lys38-GLP-1(7-38)、Arg26,34-Lys38-GLP-1(7-38)、Arg26,34-Lys38-GLP-1(7-38)、Arg26,34-Lys38-GLP-1(7-38)、Arg26,34-Lys38-GLP-1(7-38)、Arg26-Lys38-GLP-1(7-38)、Arg26-Lys38-GLP-1(7-38)、Arg26-Lys38-GLP-1(7-38)、Arg34-Lys38-GLP-1(7-38)、Ala37-Lys38-GLP-1(7-38)和Lys37-GLP-1(7-37)。
在本发明的又一实施例中,本发明肽包含有括作为组分(I)或(III)的经修饰的GLP-1肽,其包括具有下列分子式(II)(SEQ ID No.:44)的氨基酸序列:
Xaa7-Xaa8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Xaa16-Ser-Xaa18-Xaa19-Xaa20-Glu-Xaa22-Xaa23-Ala-Xaa25-Xaa26-Xaa27-Phe-Ile-Xaa3o-Trp-Leu-Xaa33-Xaa34-Xaa35-Xaa36-Xaa37,
其中,Xaa7是L-组氨酸、D-组氨酸、去氨基-组氨酸、2-氨基-组氨酸、3-羟基-组氨酸、同源组氨酸(homohistidine)、N-乙酰基-组氨酸、a-氟甲基-组氨酸、a-甲基-组氨酸、3-吡啶丙氨酸、2-吡啶丙氨酸或4-吡啶丙氨酸;Xaa8是Ala、Gly、Val、Leu、Ile、Lys、Aib、(1-氨基环丙基)羧酸、(1-氨基环丁基)羧酸、(1-氨基环戊基)羧酸、(1-氨基环己基)羧酸、(1-氨基环庚基)羧酸或(1-氨基环辛基)羧酸,特别优选Gly;Xaa16是Val或Leu;Xaa是Ser、Lys或Arg;Xaa19是Tyr或Gln;Xaa20是Leu或Met;Xaa22是Gly、Glu或Aib;Xaa23是Gln、Glu、Lys或Arg;Xaa25是Ala或Val;Xaa26是Lys、Glu或Arg;Xaa27是Glu或Leu;Xaa30是Ala、Glu或Arg;Xaa33是Val或Lys;Xaa34是Lys、Glu、Asn或Arg;Xaa35是Gly或Aib;Xaa36是Arg、Gly或Lys或氨基或缺失;Xaa37是Gly、Ala、Glu、Pro、Lys、氨基或缺失。
在本发明的又一实施例中,本发明肽的组分(I)和/或(III)包含经修饰的GLP-1肽,其包括下列分子式(III)(SEQ ID No.:45)的氨基酸序列:Xaa7-Xaa8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Xaa8-Tyr-Leu-Glu-Xaa22-Xaa23-Ala-Ala-Xaa26-Glu-Phe-Ile-Xaa30-Trp-Leu-Val-Xaa34-Xaa35-Xaa36-Xaa37,其中,Xaa7是L-组氨酸、D-组氨酸、去氨基-组氨酸、2-氨基-组氨酸、-羟基-组氨酸、同源组氨酸、N-乙酰基-组氨酸、a-氟甲基-组氨酸、a-甲基-组氨酸、3-吡啶丙氨酸、2-吡啶丙氨酸或4-吡啶丙氨酸;Xaa8是Ala、Gly、Val、Leu、Ile、Lys、Aib、(1-氨基环丙基)羧酸、(1-氨基环丁基)羧酸、(1-氨基环戊基)羧酸、(1-氨基环己基)羧酸、(1-氨基环庚基)羧酸或(1-氨基环辛基)羧酸;Xaa18是Ser、Lys或Arg;Xaa22是Gly、Glu或Aib;Xaa23是Gln、Glu、Lys或Arg;Xaa26是Lys、Glu或Arg;Xaa34是Lys、Glu或Arg;Xaa35是Gly或Aib;Xaa36是Arg或Lys、氨基或缺失;Xaa37是Gly、Ala、Glu、或Lys、氨基或缺失。
在本发明的特别优选的实施例中,本发明肽的组分(I)和/或(III)包括(经修饰的)GLP-1肽,其选自GLP-1(7-35)、GLP-1(7-36)、GLP-1(7-36)-氨基、GLP-1(7-37)或它们的变异体、类似物或衍生物。在本发明肽的组分(I)和/或(III)中,还优选地包括经修饰的GLP-1肽,该经修饰的GLP-1肽在其8位具有Aib残基或在其7位具有选自下列基团的氨基酸残基:D-组氨酸、去氨基组氨酸、2-氨基-组氨酸、羟基-组氨酸、同源组氨酸、N-乙酰基-组氨酸、a-氟甲基-组氨酸、a-甲基-组氨酸、3-吡啶丙氨酸、2-吡啶丙氨酸和4-吡啶丙氨酸。
可用于获得可在本发明中使用的类似物的生成蛋白质中氨基酸取代基的例子包括任何已知的方法和步骤,如美国专利RE 33,653、4,959,314、4,588,585和4,737,462(Mark等)、5,116,943(Koths等)、4,965,195(Namen等)以及5,017,691(Lee等),以及在美国专利4,904,584(Shaw等)中公开的赖氨酸取代的蛋白质。
优选地,以上定义的包含在组分(I)、(II)和/或(III)中的变异体或类似物,将具有中心序列,该中心序列与“天然”序列(举例来说,GLP-1(7-37)或GLP-2或它们的生物活性片段或任何IP2亚基)相同,该中心序列具有与该天然氨基酸序列至少70%同源性并保持其生物活性的氨基酸序列。更优选地,该序列与天然序列相比至少具有80%的同源性、至少90%的同源性,或更优选地至少95%的同源性。在特定的肽被认为与确定长度的参考多肽相比具有特定百分比的同源性时,这个同源性百分比是相对于该参考多肽的。因此,与100个氨基酸长度的参考多肽具有50%一致的肽可以是与参考多肽的50个氨基酸长度的部分完全一致的50个氨基酸的多肽。其也可以是100个氨基酸长度的多肽,在其整个长度中,有50%与参考多肽相同。当然,其它多肽也将符合同样的标 准。此处所使用的术语“序列同源性(sequence identity)”是指该序列进行如下比较。对该序列采用基因计算机组的GAP(总体序列对比程序(global alignment program))的版本9进行对准,采用具有-12的空格开放罚分(用于空格的第一无效值)和-4的空格延伸罚分(在该空格中的每个附加连续无效值)的默认(BLOSUM62)矩阵(值-4到11)。在经过对准后,通过将相匹配的数量表示为与所要求的序列中的氨基酸数量的百分比来计算出同源百分比。
本发明还包括融合肽的衍生物或它的类似物、片段或变异体。术语本发明肽的“衍生物”仅仅包括那些并不将一个氨基酸改变成二十个普遍存在的天然氨基酸中的另一种的经修饰的肽。相对的,可通过将其与有机衍生试剂反应来修饰基因编码的氨基酸,该有机衍生试剂能与残基的选择侧链或与终端残基的氨基酸或羧基反应(优选共价修饰);也可通过引入非天然氨基酸(化学合成制作,也就是遗传密码编码的氨基酸的二体、Aib(a-氨基异丁酸)、Abu(氨基丁酸)、Tie(叔亮氨酸)、对-丙氨酸、3-氨甲苯酸、氨基苯甲酸)来修饰基因编码的氨基酸;也可通过引入未经遗传密码编码的天然氨基酸(举例来说,羟基脯氨酸、γ-羧基谷氨盐、鸟氨酸、磷酸丝氨酸、D-丙胺酸和D-谷氨酸盐)来修饰基因编码的氨基酸;或者还可通过其它替代的肽主链排布来修饰肽主链。
下面还公开了本发明肽(如上所述,还包括融合肽的变异体、类似物或片段)的氨基酸的优选修饰,其可存在于本发明肽的任何位点(任何氨基酸),举例来说,位于组分(I)、(II)和/或(III)内。
如果半胱氨酰残基出现在本发明肽例如本发明肽的类似物的任何形态中,其一般会与α-卤代乙酸(以及其对应的胺)例如如氯乙酸或氯乙酰胺反应,以生成羧甲基或羧氨甲基衍生物。半胱氨酰残基的衍生物也可通过与溴三氟丙酮、α-溴-β-(5-咪唑基)丙酸、氯乙酰基磷酸盐、N-取代马来酰亚胺、3-硝基-2硫代吡啶、甲基-2-硫代吡啶、对-氯汞基苯甲酸盐、2-氯汞基-4-硝基酚、或氯-7-硝基苯-2-含氧酸-1,3-二唑反应而生成。
本发明肽中的组酰胺残基也可通过在pH 5.5-7.0与二乙基焦碳酸酯反应衍生,因为该试剂对于组酰胺侧链有相关特效。溴苯甲酰甲基溴也是有用的,该反应优选在pH 6.0的0.1M的二甲胂酸钠中进行。特别,本发明肽的组分(I)和/或(III)中包含的GLP-1(7-37)的N-端组酰胺残基(His)对GLP-1肽的促胰岛素活性非常重要,参见Suzuki等所示(Diabetes Res.;Clinical Practice 5(Supp.1):S30(1988))。相对地,可采用烷基或酰基(C1-C6)基团在作为组分(I)和/或(III)的GLP-1的His7,对本发明肽进行修饰,或采用功能等效的C5-C6环状结构取代His。较佳的修饰是在His7的氨基末端或其组酰胺侧链上引入疏水基。
举例来说,本发明肽的赖氨酰和氨基末端残基可与丁二酸酐或其它羧酸酐反应。与这些试剂的衍生反应具有改变赖氨酰残基电荷的功效。通过对本发明肽的赖氨酰残基的ε-氨基酸基团的酰基(C12-C18)修饰,其在循环中的半衰期增长了。举例来说,可通过与一种或多种传统试剂反应修饰精氨酰残基,这些试剂包括苯甲酰甲醛和茚三酮。由于胍官能团的高pKa,精氨酰残基的衍生需要在碱性环境中进行。此外,这些试剂可与赖氨酸基团以及精氨酸的ε-氨基酸基团反应。
酪氨酰残基的特定修饰的本质已经得到了广泛的研究。最常用地,N-乙酰基咪唑和四硝基甲烷可用于分别形成O-乙酰基酪氨酰族和e-硝基衍生物。
通过将羧基侧基(天门冬氨酰或谷氨酰)与碳二亚胺(R′N-C-N-R′)例如1-环乙基-3-[2-马啉基]-(4-乙烷基)碳二亚胺或1-乙烷基-3-(4-氮阳离子)-4,4-二甲基苯)碳二亚胺进行反应,可以对羧基侧基进行修饰。
此外,通过与铵离子反应,可将天门冬氨酰和谷氨酰转换成天冬酰胺酰和谷氨酰胺酰残基。
对天冬酰胺酰和谷氨酰胺酰残基进行脱酰胺作用,可以生成对应的天门冬氨酰和谷氨酰。可替换地另一情况下,可在弱酸条件下对这些残基进行脱酰胺作用。这些残基的任何形态均落入本发明的保护范围。脱掉酰氨基的本发明肽对蛋白酶或肽酶的蛋白质水解的敏感性可能发生改变,表明脱酰胺作用在本发明肽的蛋白水解中具有生理重要性。注意,生物合成的本发明肽可在某种存储条件下降解,导致本发明肽中的一个或多个位点上的出现脱酰胺反应。本发明肽中的蛋氨酸残基易于被氧化成亚砜。如上面所提到的其它衍生物,本发明脱掉酰氨基的肽和/或亚砜肽均可具有完整的生物活性。
其它适用于衍生包含有α-氨基酸的残基的试剂包括酰亚胺酯,如甲基吡啶三氟脒菊酯(methyl picolinimidate)、吡哆醛磷酸盐、吡哆醛、硼氢化氯、三硝基苯磺酸、O-甲基异脲(O-methyliosurea)、2,4-间戊二酮;并由转氨酶催化与乙醛酸反应。
使用合适的氨基或羧基保护基团,本发明肽的末端氨基酸残基的羧基(C-端)和氨基(N-端)基团可以以被保护(举例来说,用氨基基团保护其C-端)或未被保护的形态出现。也可提供本发明肽的酸加成盐。常见的酸加成盐是氢卤酸盐,也就是,HBr、HI,HCl,其中HCl较佳。
在本发明肽中出现的末端、侧链羧基基团、或赖氨酸的ε-氨基基团的聚乙二醇化作用,可为本发明肽提供抗氧化性,并也落入本发明的保护范围。
导致本发明肽的衍生反应的其它的修饰均基于可与本发明肽共价相连的碳水化合物和/或油脂。首选将油脂和/或碳水化合物通过其活性侧链基与丝氨酸、苏氨酸、天冬酰胺酸、谷氨酸盐、酪氨酸、谷氨酸酯或天冬酰胺酸盐相连。或者,油脂和/或碳水化合物也可与本发明肽的末端基团相连。此外,本发明肽可与不同的官能肽或蛋白质基团相连,这些官能肽或蛋白质基团可稳定本发明肽和/或用于改进本发明肽在体液特别是血液中的传输特性。举例来说,可从清蛋白、铁转移蛋白等中选择合适的肽或蛋白质,这些肽或蛋白质可与本发明肽直接相连,或通过肽或有机接头序列与本发明肽相连(作为组分IV)。优选将这些肽或蛋白质连接在本发明肽的一端。
为了解决本发明肽的降解的难题,本发明的又一实施例提供了一种由D氨基酸或至少部分由D氨基酸组成的本发明肽的逆反异构体(retro-inversoisomer)。术语“逆反异构体”是指线性肽的异构体,其中序列的方向是逆反的,并且每个氨基酸残基的手性是反向的(参见Jameson et al.,Nature,368,744-746(1994);Brady et al.,Nature,368,692-693(1994))。相对于亲本肽,该逆反异构肽是以氨基酸的反序组装的,一般具有F-moc氨基酸衍生物。特别地,该未加工的肽可由反向HPLC纯化。
其他可引入到本发明肽中的修饰与肽骨架的修饰有关。优选地,修饰后的本发明肽是支架拟态(scaffold mimetics)。它们的骨架不同于天然存在的骨架,但是它们的侧链与本发明肽或其片段、变异体、衍生物或类似物相同。通常,支架拟态呈现了对一个或多个主链成员(NH,CH,CO)的修饰,可以是通过取代(优选)或插入。举例来说,取代是(I)-O-、-S-或-CH2-取代-NH-;(II)-N-、C-Alkyl-或-BH-取代-CHR-;(III)-CS-、-CH2-、-SOn-、-P=O(OH)-或-B(OH)-取代-CO-.。本发明肽的模拟肽可以是这些修饰中的每一种修饰的结合。特别地,可将每个基团I、II和III的修饰相结合。在模拟肽中,可对每个主链成员进行修饰,或者采用非天然生成的基团取代一定数量的主链成员。优选地,本发明肽中的所有的主链成员:-NH-、-CHR-或CO,均被另一非天然生成的基团取代。在本发明肽骨架的酰胺键(-NH-CO-)被取代的例子中(在整个分子中或至少在单个位点),较佳的取代基是生物等排电子,举例来说,逆反酰胺键(-CO-NH-)、羟乙烯基(-CH(OH)-CH2-)、烯烃(CH2=CH-)、卡巴结构(carba)(CH2-CH2-)和/或-P=O(OH)-CH2-)。或者,通过插入实现的主链延长可在本发明肽的模拟支架中发生,举例来说,通过侧接在C-α原子的基团。举例来说,可在C-α原子的任一侧插入-O-、-S-、-CH-、-NH-。
本发明肽的低聚氨基甲酸酯(oligocarbamate)肽骨架结构为最佳。其酰胺键由氨基甲酸基取代。可通过相应的氨基酸或氨基醇获得N-端保护的氨基碳酸烷基酯单体。通过固相合成法,使用F-moc基或光敏硝基芳氧羟基团(nitroatryloxycarbonyl group),可将这些单体转换成活性酯,例如p-硝基苯酯。
对本发明肽进行如上保护以防止其蛋白水解。特别对其进行保护以抗二肽氨基肽酶-4(DPP-IV)。在此使用的术语“(DPP-IV)保护”是依照指权利要求1所述的肽。本发明肽以及其衍生物、类似物、片段和变异体均将GLP-1(7-35、36或37)作为其组分(I)和/或(III)的一部分,并使其对抗血浆肽酶(DPP-IV)具有抗性。
肽的抗二肽氨基肽酶-4(DPP-IV)降解特性可通过以下降解实验证明。将肽的等分试样和纯化的抗二肽氨基肽酶IV的等分试样在适合的缓冲溶液(缓冲溶液不能是清蛋白)中孵化4-22小时(实验条件:37摄氏度,pH 7-8)。加入三氟醋酸后,酶促反应被终止,使用HPLC或LC-MS分析将肽的降解产物分离纯化。执行该分析的一种方法是:将混合物运用到Zorbax 300SB-C18(30nm孔径,5μm微粒)150×2.1mm,并在0.1%的三氟醋酸(0%-100%乙腈,超过30min)中采用0.5ml/min的流速且具有乙腈线性梯度的条件下洗脱。在214nm(肽骨架)或280nm(芳香氨基酸)观察肽和其降解产物的吸收率,并由其峰值面积的积分对其定量。使用LC-MS确定其降解形式,在此,可确定分离峰的MC谱。在特定时间的原化合物/降解化合物的百分比用于估计肽的DPP-IV稳定性。
当在给定的时间里,本发明肽的原化合物所占的百分比比GLP-1(7-37)稳定10倍时,可将其定义为DPP-IV稳定。这样,DPP-IV稳定的本发明肽至少比GLP-1(7-37)稳定10倍,更佳地比GLP-1(7-37)稳定20倍。对于本领域技术人员,可通过任何已知的方法评估肽的稳定性,举例来说,通过在将要进行检测的肽溶液中加入DPP-IV,采用分光物镜法、免疫印迹分析法、抗体筛选法来通过确定肽随着时间的推移的降解。同时的,本发明肽被定义为化合物,其通过与GLP-1(7-37)的天然受体(GLP-1(7-37)受体)绑定来发挥GLP-1(7-37)的功效。优选地,本发明肽对GLP-1(7-37)受体具有亲和力,其具有至少10%、优选至少50%的对应的天然存在的GLP-1肽的亲和力。可采用任何适合的方法判断这种亲和力,举例来说,表面等离子体共振等。此外,更佳的是,本发明肽通过其与细胞外受体的绑定,将信号传递到细胞内,并唤醒细胞内cAMP的形成。
可使用固相肽合成技术合成本发明的肽,合成过程类似与本领域中合成GLP-1(7-36)醯胺和GLP-1(7-37)的方式。然后,本发明肽可以实验室规模纯化,举例来说,在逆向HPLC色柱上的一步纯化或使用适合的色谱法。
然而,更适合在工程细胞(微生物细胞或动物细胞系)中生成本发明肽。本发明肽可以从其表达的细胞中分离,例如,采用传统的分离技术。这样细胞可在合适的条件下(例如包括支撑物和养分的)在试管中成长并分泌蛋白,也就是,从细胞外基质中重新获得本发明肽。这样,引入到细胞中的序列优选地包括本发明肽的前导序列(leader sequences)和信号肽序列。优选地,该细胞表达可裂解该前导序列和信号序列的蛋白酶,这些序列可以是内生基因序列或工程基因序列。在一个可替换的示例中,编码本发明肽的工程基因序列并不包括该前导序列和信号肽序列,因此将不会分泌细胞内表达的本发明肽,而通过处理相关的细胞溶菌重新从细胞获得本发明肽。在这些方法中,编码的序列可包括纯化标记,该标记使得可从媒介中有效地萃取产物肽,也可裂解该标记以释放单个的本发明肽。
本发明进一步提供了一种核苷酸,该核苷酸用于编码本发明肽,使其成为融合肽或它的片断、类似物或变异体。本发明包括任何编码发明肽的核苷酸。由于遗传密码的兼并性,多种核苷酸序列可用于编码发明肽。落入本发明保护范围的核苷酸分子也可包含编码发明肽的核苷酸,另外,更进一步地(功能性地)包含核苷酸序列。在本发明的优选实施例中,这样一个核苷酸分子可编码(a)整个GLP-1aa序列(GLP-1(1-37)或功能的GLP-1(7-35、36或37)序列,(b)位于依照(a)的GLP-1序列的N-端的裂解序列,该裂解序列可用于任何蛋白质,(b)的上游可用于编码前导序列。在另一优选实施例中,编码(b)核苷酸分子的核苷酸序列上游可另外包括(c)编码信号肽的序列。或者,本发明核苷酸分子可具有融合上游(a)的序列(c),但其间并不包括任何编码前导序列(b)的序列。优选地,对于胰高血糖素前原(preproglucagon),前导序列和信号肽序列是异源的。
本发明肽进一步提供一种媒介,其包括本发明核苷酸(分子)和其它用于本发明核苷酸(分子)的表达的功能组分。一般地,本发明核苷酸(分子)将和启动子序列融合,并最终与其它调节序列(如增强子序列)组合。对于复制,质粒可包括复制起点。为了选择可转染本发明媒介的细胞,可在该媒介中提供一个或多个抗生素抗性基因(举例来说,卡那霉素、安匹西林)。该媒介可以是质粒,其包括用于细菌原始启动子、抗生素抗性基因、和在哺乳动物细胞中复制和表达的抗生素抗性基因和原始启动子。本发明进一步提供一种宿主细胞,该细胞包括外源引入的本发明的DNA,可用于转译所述前体蛋白质。该宿主细胞可以是原核宿主细胞或真核宿主细胞,举例来说,哺乳动物细胞。
根据本发明的另一方面,提供了一种用于动物的治疗方法,优选用于人类的治疗方法,该方法通过施用包括组分(I)和(II)以及组分(III)的本发明肽来实现。本发明还提供了本发明肽在制造用于治疗或预防与葡萄糖代谢有关的疾病或不适的产品中的应用。葡萄糖紊乱的非限制性例子包括:I型糖尿病、II型糖尿病(NIDDM)、胰岛素抗性、体重失调或与之相关的疾病或不适,其中体重失调或相关不适包括肥胖、超重引起的不适、反常过饱、血浆胰岛素水平降低、血糖浓度升高或胰腺β细胞量减少。优选地,在此公开了发明肽在制造用于治疗II型糖尿病(NIDDM)的药剂中的应用。因此,本发明涉及本发明肽的使用,举例来说,用于降低受治疗者的体重,用于减轻受治疗者的过饱感,用于升高受治疗者餐后的血浆胰岛素水平,用于降低受治疗者禁食的血糖水平,用于增加受治疗者的胰腺β细胞量,或用于治疗受治疗者的I型或II型糖尿病。
本发明肽(也就是,融合肽或其类似物、片段、变异体或衍生物等)可治疗患有其它疾病或生理紊乱的病人。本发明肽可用于制作药剂,这些药剂可用于治疗神经退行病变(neurodegenerative disorders)以及与此相关的疾病或生理状况,还可用于治疗与下垂症相关的紊乱、疾病或生理状况。本发明肽对这些紊乱的治疗的使用取决于下列因素:与环AMP第二信使途径相连的GLP-1受体,该受体可在啮齿动物和人类的脑髓中完全表达。在脑髓中受体分布的化学构建(chemoarchitecture)并不仅仅只与在日常食物摄取中的GLP-1的中心作用有关,其还响应有害物应力(aversive stress)。应了解,与其GLP-1受体绑定的GLP-1发挥神经营养特性,并在培养的神经元细胞中提供抗谷氨酸盐诱导的下垂症和抗氧化伤害。此外,已显示出GLP-1对细胞培养中的淀粉状β-蛋白前体处理进行的修饰,并且一定剂量的GLP-1可降低在体的脑髓内的淀粉状β-蛋白水平。因此,已知GLP-1是中枢神经系统的调节者。模拟生理活性的GLP-1的生物活性的本发明肽可用于治疗例如阿尔茨海默病(AD)和其它中枢和边缘神经退行性疾病(举例来说,肌萎缩性脊髓侧索硬化症(ALS)、亚历山大病、阿耳珀氏病、共济失调毛细管扩张症、脑白质海绵状变性、柯凯因氏症候群、克雅氏病、多发性硬化、桑德霍夫病、皮克氏病、脊髓小脑性共济失调、谢尔德氏病和帕金森病)。
此外,已示出生理活性的GLP-1可在不同的细胞上发挥抗细胞凋亡的功效,举例来说,GLP-1有益于新鲜的单个人类胰岛细胞或其它类型的细胞团和细胞功能的保护。在此范围内,该生物活性的本发明肽可用于治疗由细胞或组织凋亡引起的紊乱。
本发明肽可用于制造一种可外部给予的化合物,其包括本发明肽。生成的化合物也可用于治疗上述疾病。此处公开的疾病也可通过本发明宿主细胞、核苷酸(分子)或媒介来治疗,甚至,本发明宿主细胞、核苷酸(分子)或媒介可用于制备可治疗这些疾病的药剂。
包含本发明肽序列作为活性成分的制备剂型为本领域所熟知,例如美国专利4,608,251、4,601,903、4,599,231、4,599,230、4,596,792和4,578,770,在此将这些专利引入以作参考。通常,这些剂型可制作成血管注射剂,如流体或悬浮液,较佳地包括水(水合剂型)或可将其乳化。术语“水合剂型”定义为至少包括50%w/w水。类似的,术语“水合溶液”定义为至少包括50%w/w水的溶液,且术语“水合悬浮液”定义为至少包括50%w/w水的悬浮液。
对于静脉注射、皮肤注射或皮下注射,或在患处的注射,该活性成分可采用肠道外可接收水合溶液的形式,其是无热原的,并具有适合的pH、等渗力和稳定性。液体药物组分一般包括流体媒介,如水。优选地,该流体媒介将包括生理盐溶液、葡萄糖乙醇,或其它糖溶液,或乙二醇,如乙烯基乙二醇、丙二醇或聚乙二醇,或者该流体媒介可包括这些溶液和组合。进一步的例子是其它的等渗媒介,如林格注射剂或乳酸盐林格注射剂。
如果本发明涉及的药物剂型包括依照本发明的化合物的水溶液和缓冲剂,其中所述化合物的浓度为0.1mg/ml或更高,且其中所述剂型的pH范围从2.0到10.0,更佳地从7.0到约8.5。优选地,该剂型离根据本发明的化合物的等电位点至少是1pH单位,更优选地,该剂型离根据本发明的化合物的等电位点至少是2pH单位。
也可制备在注射以前可溶解或悬浮在液体中的固体形式。该药物剂型可以是冻干剂型,因此在使用之前,医师或病人可在其中添加溶液和/或稀释剂。换句话说,一旦该剂型制备好了,不必马上给患者施用。相反的,制备好后,将其包装好在冰冻状态下保存,或在干燥状态下保存并可稍后补充试剂以成为液体状态,或以其它适合患者使用的形式。在另一实施例中,该药物剂型是干燥剂型(举例来说,冻干或喷干),其可直接使用无需任何预先溶解。通过“干燥形式”是指通过冷冻干燥(也就是,冻干法,参见如Williams和Polli(1984)J.Parenteral Sci.Technol.38:48-59)、喷射烘干(见Masters(1991)inSpray-Drying Handbook(5th ed;Longman Scientific和Technical,Essez,U.K.),pp.491-676;Broadhead等(1992)Drug Devel.Ind.Pharm.18:1169-1206;和Mumenthaler等(1994)Pharm.Res.11:12-20)或空气干燥(Carpenter和Crowe(1988)Cryobiology 25:459-470;和Roser(1991)Biopharm.4:47-53)该液体药物成分或剂型。在液体药物组分的储存中,团聚体的形成可能逆向影响该多肽的生物活性,导致该药物组分治疗功效的降低。此外,团聚体的形成可导致其它的问题,如当包括多肽的药剂组分是使用注射系统给与时,可能造成管道、膜或泵的堵塞。
在本发明的肽药剂剂型中,可能存在其它的试剂。这些附加试剂可包括湿润剂、乳化剂、抗氧化剂、填充剂、pH缓冲剂(举例来说,磷酸、柠檬酸盐或马来酸盐缓冲液)、防腐剂、表面活性剂、稳定剂、弹性改性剂(tonicitymodifiers)、金属离子、螯合剂、脂性媒介、蛋白质(举例来说,人血清蛋白、凝胶或蛋白质)和/或两性离子(举例来说,氨基酸,如甜菜碱、牛磺酸、精氨酸、甘氨酸、赖氨酸和组氨酸)。
至于本发明剂型的稳定剂,优选那些高分子量聚合物或低分子化合物。在本发明的进一步的实施例中,该稳定剂可选自:聚乙二醇(举例来说,PEG3350)、聚乙烯醇(PVA)、聚乙烯吡咯烷酮、羧基氧化纤维素(carboxy-hydroxycellulose)或它的衍生物(举例来说,HPC、HPC-SL、HPC-L和HPMC)、环式糊精、含硫磺的基质(如硫代甘油、2-硫基乙醇和2-甲基硫代乙醇(2-methylthioethanol))和不同的盐类(举例来说,氯化钠)。这些特定的稳定剂中的任意一个组成本发明的一个实施例。该药剂组分还可包括附加稳定剂,其可进一步增强此处的治疗活性多肽的稳定性。本发明的特别重要的稳定剂包括但不限于蛋氨酸和EDTA,其可保护多肽使其抗蛋氨酸氧化,以及非离子表面活性剂,其可保护多肽使其抗与冻融或机械剪切相关的聚集。
至于本发明剂型的表面活性剂,可较佳地选自清洁剂、乙氧基蓖麻油(ethoxylated castor oil)、聚乙二醇甘油酯、乙酰化单甘酯、山梨糖醇酐脂肪酸酯、聚氧乙烯-聚氧丙烯多嵌段聚合物(举例来说,泊洛沙姆,如泊洛沙姆液体(Pluronie)F68、泊洛沙姆188和407、曲拉通100)、聚氧丙烯山梨糖醇酐脂肪酸酯、成形PEO、聚氧乙烯和聚氧丙烯衍生物(如烷基化物或烷基化衍生物(吐温,举例来说,吐温-20、吐温-40、吐温-80和Brij-35))、聚氧丙烯羟基硬酯酸、甘油一酸酯或其乙氧基化衍生物、甘油二酯或其聚氧丙烯衍生物、乙醇、甘油、卵磷脂、磷脂(举例来说,磷脂酰丝氨酸、磷脂酰胆碱、磷脂酰乙醇胺、磷脂酰纤维醇、双磷脂酰甘油和鞘磷脂)、磷脂衍生物(举例来说,二棕榈酰基磷酸酯(dipalmitoyl phosphatidic acid))和溶血磷脂(举例来说,棕榈酰基溶血磷脂酰-L-丝氨酸(palmitoyllysophosphatidyl-L-serine)和乙醇胺的1-酰基-甘油-3-磷酸酯、胆碱、丝氨酸或苏氨酸)、以及溶血磷脂和卵磷脂的烷基、烷氧基(烷基酯)、烷氧基(烷基酯)衍生物(举例来说,溶血磷脂和二棕榈酰基磷酸胆碱的月桂酰和肉豆蔻酰衍生物)、以及极性头端(也就是,胆碱、乙醇胺、磷脂酸、丝氨酸、苏氨酸、甘油、纤维醇,以及正电荷的DODAC、DOTMA、DCP、BISHOP、溶血磷脂酰丝氨酸和溶血磷脂酰苏氨酸)、以及环甘油磷脂(举例来说,脑磷脂)、环甘油醣脂(举例来说,半乳糖苷(galactopyransoide))、鞘氨醇醣脂(举例来说,神经酰胺、神经节苷脂)、十二烷基胆碱磷酸、鸡蛋溶血卵磷脂、梭链孢酸衍生物(举例来说,牛磺-梭链孢酸钠等)、长链脂肪酸及其盐C6-C12(举例来说,油酸和辛酸)、酰基肉碱、赖氨酸、精氨酸或组氨酸的N`X-酰化衍生物、或赖氨酸的侧链酰化衍生物、或缩二肽的精氨酸N-酰化衍生物(该缩二肽包括赖氨酸、精氨酸或组氨酸的任意组合和一种中性或酸性氨基酸)、三肽的N-酰化衍生物(该三肽包括一种中性氨基酸和两种带电荷的氨基酸的组合)、DDS(多库酯钠(docusatesodium),CAS登记号[577-11-7],多库酯钙,CAS登记号[128-49-4],多库酯钾,CAS登记号[7491-09-0])、SDS(十二烷基硫酸钠或十二烷醇硫酸钠)、辛酸钠、胆酸及其衍生物、胆汁酸及其盐和甘氨酸或牛磺酸的共轭化合物、熊去胆酸、胆酸钠、脱氧胆酸钠、牛磺胆酸钠、甘胆酸钠、N-十六烷基-N、N-二甲基-3-氨-1-丙烷磺酸盐、单价阴离子(烷基-芳基-磺酸盐)表面活性剂、两性离子表面活性剂(举例来说,N-烷基-N,N-e二甲氨基-1-丙烷磺酸盐、3-胆汁氨-1-丙烷二甲氨基-丙烷-磺酸盐)、阳离子表面活性剂(季铵基)(举例来说,十六烷基-三甲基溴化铵、氯化十六烷基吡啶)、非离子型表面活性剂(举例来说,十二烷基-D-葡萄糖苷)、泊洛沙胺(举例来说,曲拉通,该泊洛沙胺是衍生自向乙撑二胺中连续加入环氧丙烷和乙撑氧的四功能化合物的嵌段共聚物)。或者表面活性剂可从咪唑啉衍生物或混合物选取。这些特定表面活性剂中的每个都可组成本发明的可选实施例。对本领域技术人员来说,表面活性剂在药物组分中的应用是众所周知的(可参考Remington:The Science and Practice ofPharmacy,19th edition,1995)。
关于配药学上可使用的防腐剂较佳地选自下列物质:苯酚、o-甲酚、m-甲酚、p-甲酚、甲基p-羟基苯甲酸、丙基p-羟基苯甲酸、2-苯氧基乙醇、苯甲醇、乙醇、氯丁醇,以及硫柳汞、溴硝醇、苯甲酸、咪唑烷脲、氯己啶、脱氢醋酸钠、氯化甲酚、乙烷基p-羟基苯甲酸、乙氧胺氯(benzethonium chloride)、氯苯甘醚(3p-氯苯氧基丙烷-1,2-二醇)或它们的混合物。
关于等渗剂,可较佳地选自下列物质:盐(举例来说,氯化钠)、糖或糖醇、氨基酸(举例来说,L-甘氨酸、L-组氨酸、精氨酸、赖氨酸、异亮氨酸、天门冬氨酸、色氨酸、苏氨酸)、醛醇(举例来说,甘油(glycerol)(丙三醇(glycerine))、1,2-丙二醇(丙二醇(propyleneglycol)、1,3-丙二醇、1,3-丁二醇)聚乙二醇(举例来说,PEG400)、或它们的混合物。可使用任何糖类:如单糖、二糖或多糖、或水溶性葡聚糖,包括如果糖、葡萄糖、甘露糖、山梨糖、木糖、麦芽糖、乳糖、蔗糖、海藻糖、右旋糖苷、茁霉多糖、糊精、环式糊精、可溶淀粉、羟乙基淀粉、羧甲基纤维钠。在一个实施例中糖类添加剂是蔗糖。糖醇是定义为至少具有一个-OH基的C4-C8烃,其包括如甘露醇、山梨醇、纤维醇、半乳糖醇、卫矛醇、木糖醇和阿糖醇。在一个实施例中,该糖醇添加剂是甘露醇。上述的糖类或糖醇可以单独使用或组合使用。对糖类或糖醇用量没有固定的限制,只要它们可溶解在准备液体中,且不会影响采用本发明的方法获得的稳定效果即可。
关于螯合剂,可较佳地选自下列物质:乙二胺四乙酸(EDTA)的盐、柠檬酸和天门冬胺酸,以及它们的混合物。
关于缓冲溶液,可较佳地选自下列物质:乙酸钠、碳酸钠、柠檬酸盐、双甘氨肽、组氨酸、甘氨酸、赖氨酸、精氨酸、磷酸二氢钠、磷酸氢二钠、磷酸钠和三羟甲基氨基甲烷、羟乙基哌嗪乙磺酸(hepes)、N-二羟乙基甘氨酸、蓖麻毒、苹果酸、琥珀酸盐、马来酸、反丁烯二酸、酒石酸、天门冬胺酸,以及它们的混合物。这些特定缓冲液中的任意一种可组成本发明的可选实施例。
对本领域技术人员来说,在包含本发明用于治疗的肽的配药组分中的所有的前述的添加剂的使用是众所周知的,特别是浓度范围是已知的。为便于理解,可参考Remington:The Science and Practice of Pharmacy,19th edition,1995。
包含本发明肽的剂型一般不经肠道给予,通过注射,例如皮下注射(subcutaneously)、皮内注射(intradermally)、真皮下层注射(subdermally)或肌肉注射。本发明肽的肠胃外投药的化合物的制备可参见WO 03/002136。
其它的适合其它给予方式的剂型包括栓剂,且在某些情况下,栓剂可口服或放置在面颊内、舌下、腹膜内、叶鞘内、肛门内、头颅骨内。对于栓剂,传统的粘合剂和载体可以包括例如聚乙烯乙二醇或甘油三酸酯。这样的栓剂可由包含0.5%到10%的活性组分的混合剂制成,活性成分的含量优选12%。口服剂型包括常用的赋性剂,如制药级别的甘露醇、乳糖、淀粉、硬脂酸镁、糖精钠、纤维素、碳酸镁以及类似物。这些组分采用溶液、悬浮液、药片、药丸、胶囊、持续释放药剂或药粉的形式,包含10-95%的活性试剂,优选25-70%。
如上所述,可采用额外的药剂方法来控制药效的持续时间。通过使用聚合体来复合或吸收本发明的肽,可控制释放作用。活性成分(肽)的控释释放可通过选择适合的高分子(如,聚酯、聚氨基酸、聚乙烯吡咯烷酮、乙烯-醋酸乙烯聚合物、甲基纤维素、羧甲基纤维素、鱼精蛋白硫酸盐)、高分子的浓度、以及结合的方法来实现。在Remington′s Pharmaceutical Sciences(见上述)中已经公开了此类教导。控制药效持续时间的另一可行的方法是将本发明的肽结合到聚合材料(如聚酯、聚氨基酸、水凝胶、聚(乳酸)或乙烯-醋酸乙烯聚合物)颗粒中。
本发明肽可制作成中性或盐形式。本发明的肽可以与多种有机和无机碱、多种有机和无机酸中的任何一种反应,以形成(加成)盐(举例来说,与肽的自由氨基基团反应)。常用的形成酸加成盐的酸可以是无机酸,如盐酸、溴化氢、溴化碘、硫酸、磷酸以及类似的酸;有机酸,如酒石酸、天冬氨酸-甲苯磺酸、甲烷磺酸、草酸、扁桃酸、p-溴苯基-磺酸、碳酸、琥珀酸、柠檬酸、苯甲酸。这些盐的例子包括硫酸盐、焦亚硫酸盐[、硫酸氢盐、亚硫酸盐、重亚硫酸盐、磷酸盐、磷酸氢盐、磷酸二氢盐、偏磷酸盐、焦磷酸盐、氯化物、溴化物、碘化物、醋酸盐、丙酸盐、癸酸盐、辛酸盐、丙烯酸盐、甲酸盐、异丁酸盐、己酸盐、庚酸盐、丙炔酸盐、草酸盐、丙二酸盐、琥珀酸盐、辛二酸盐、癸二酸盐、延胡索酸盐、马来酸盐、丁炔-1,4-二元酸盐、乙炔-1,6-二元酸盐、苯甲酸盐、氯苯甲酸盐、硝基苯甲酸盐、羟基苯甲酸盐、甲氧基苯甲酸盐、苯二甲酸盐、硫酸盐、二甲苯硫酸盐、乙酸苯盐、丙酸盐、苯基丁酸盐、柠檬酸盐、乳酸盐、γ-羟丁酸盐、乙二醇盐、酒石酸盐、甲烷硫酸盐、丙烷硫酸盐。自由碳基团形成的盐可衍生自无机碱,如钠、钾、铵、钙或氢氧化铁,以及有机碱,如异丙胺、三甲胺、2-乙氨基乙醇、组氨酸、普鲁卡因及其类似物。本发明肽的酸加成盐、羧酸盐、低烷基酯和醯胺均可根据WO 91/11457(1991)、EP 0 733 644(1996)和U.S.5,512,549(1996)制备。
包含本发明肽序列的药剂可以一定剂量的形式给与,并且在这样一个剂量具有治疗效果。给与的剂量取决于受治疗者,包括如病人的疾病的严重程度。合适的剂量范围是,举例来说,达到每个治疗剂量含几百微克的活性成分,活性成分的优选范围是0.1μg到2000μg(即使在1-10mg范围内的更高的量,也是可行的),如其范围可以是从0.5μg到1000μg,更佳的范围是从1μg到500μg,最佳的范围是从10μg到100μg。
包含本发明肽的并具有附加赋形剂如甘氨酸和甘露醇或其它添加剂的药剂,可装在小瓶中以冻干的形式出售。成套稀释药水瓶的提供,允许病人在服药以前,将产品重新配置成理想的浓度。本发明药剂也可以以其它已知的方式在市场上出售,如预先填充的注射器等。
本发明将在后续实施例中进一步给予介绍。
具体实施方式
实施例1
基因结构的创建
GLP-1(7-37)cDNA的编码序列是在包括Hincll和EcoR1位点的序列中综合合成的,如图1a所示。图1b中分别示出的cDNA是合成的,如图1b所示其包括GLP-1(7-37)的编码序列、IP2和Sfo1、EcoR1和Xba1的限制位点。为了使GLP-1集中在分泌途径,使用了溶基质素3(Acc.No.NM 005940)的异源编码序列。因此,编码溶基质素信号和前导序列的cDNA是从人类DNA逆转录酶PCR(聚合酶链反应)扩增的,并使用图1a或1b示出的结构分别形成图1c和1d中示出的结构。
图1a结构中的Hincll和EcoR1片段被克隆到图1d的序列的SfoI位点以形成图1e的结构。类似地,图1d的EcoR1片段被克隆到真核表达质粒的EcoR1位点,以生成图1f中示出的结构。为了形成图1g中示出的结构,图1b中示出的HincII/XbaI片段被分别克隆到图1d中示出的结构的SfoI/XbaI位点。图1h示出了编码该溶基质素前导和信号序列的合成遗传密码优化的序列,该序列被缩短的内源性内含子序列阻断,与编码人类GLP-1(7-37)、IP2和GLP-2(1-35)的序列融合。结构图1h的DNA序列是SEQ ID No.:16,而SEQ IDNo.:15也示出了该转译肽的序列。
图1i和1j中的序列也是合成的。接着,通过将图1j的Nael/BssHII克隆到图1h的Nael/BssHII线状序列,可形成图1k中的结构。结构图1k的DNA序列是SEQ ID No.:14,而SEQ ID No.:13也示出了该转译肽的序列。图1l的结构是由图1h的序列的BssHll消化和重接修复构成的。结构图1l的DNA序列是SEQ ID No.:18,而SEQ ID No.:17也示出了该转译肽的序列。Fig.1m的结构是通过将Fig.1i的序列的Afel/BssHII片段克隆到图1h的Afel/BssHII线性序列而形成的。结构图1m的DNA序列是SEQ ID No.:20,而SEQ ID No.:19也示出了该转译肽的序列。
本领域技术人员可采用常用的方法完成上述结构。
实施例2
哺乳动物细胞的转染、克隆选择和GLP-1表达
细胞源:HEK293(人类胚肾细胞系,#ACC 305,DSMZ,细胞培养采集,德国)、AtT20(鼠LAF1脑下垂体瘤细胞系,#87021902,欧洲细胞培养采集,英国),hTERT-MSC细胞是由丹麦欧登塞大学医院的Kassem教授培养的。
将具有不同GLP-1结构的0.5-2μg质粒DNA用于转染106细胞。这些GLP-1结构的合成如实施例1中所示。按照分子生物学实验方法汇编(Ausubelet al.1994ff Harvard Medical School Vol2.,Unit 9.1)中所述的方法,采用标准磷酸钙共沉淀法转染HEK293细胞。按照分子生物学实验方法汇编(Ausubel et al.1994ff Harvard Medical School Vol2.,Unit 9.1)中所述的方法,采用非脂质体基因转染试剂(FuGene)(罗氏公司)转染AtT20细胞。采用非病毒基因转染技术完成hTERT-MSC细胞的转染,该技术是基于电学参数和细胞类型特定溶剂的组合的非病毒型方法。使用非病毒基因转染设备(程序C17)和非病毒基因转染溶液VPE-1001,将获得>60%的转染效率。48小时后,通过在培养介质中加入选择性试剂杀稻瘟菌素(2μg/ml),执行细胞克隆体的转染选择,且DNA稳定整合入生物染色体。12-15天后,可将稳定的转染细胞克隆体离析并延展塑造。
可在hTERT-MSC和HEK293细胞中测量不同GLP-1结构的瞬时表达。虽然在单显性GLP-1结构#103和#317(仅仅具有GLP-1(7-37)的一个复制体)中仅可找到边缘活性(marginal active)GLP-1水平,在hTERT-MSC和HEK293细胞的二聚GLP-1结构#217(具有作为组分(I)和作为组分(III)的GLP-1(7-37))中可获得表达的聚大增益。图2中概述了结果。该结构到GLP-1结构#159(具有4个IP2复制体作为组分(II))的延伸将不会导致进一步的明显的增加(未示出)。在具有不同结构克隆体的hTERT-MSC细胞的转染选定后,其将稳定地表达GLP-1。该表达水平在表1中示出。
表1
结构 | 细胞克隆体 | 每小时且每106细胞的活性GLP[]pmol] |
#103_GLP1(7-37) | 49TM113/13 | 0,4 |
#317_GLP1(7-37)-IP2-11aa | 71TM169/1 | 0,3 |
#217_GLP1(7-37)-IP2-GLP1(7-37) | 79TM217/13 | 2,7 |
实施例3
从哺乳动物细胞分泌的GLP-1肽的蛋白印迹分析(Western Bolt Analyst)
将来自GLP-1分泌细胞的细胞培养悬浮液分散在10%-20%浓度梯度的SDS PAGE(120V,90分钟)中,并通过半干引印迹转移槽(2.0mA/cm2,60分钟)将其转移到PVDF膜(转印膜0.45μm微孔IPVH 00010)。在甲醇固定和阻断(3%(w∶v)BSA,TBS中的0.1%(v∶v)吐温-20)后,在4℃ o/n使用1μg/ml抗-GLP-1抗体(HYB 147-12,Antibodyshop)免疫印迹该膜。在RT使用0.02μg/ml检测抗体(抗鼠IgG、HRP共轭、珀金埃尔默PC2855-1197)洗涤和孵化4小时后,荧光检测将显示该蛋白质的位置。
图3中示出了蛋白印迹分析(1:在空载体转染hTERT-MSC细胞的悬浮液中溶解100ng合成GLP-1(7-37),2:从结构#217分泌二聚GLP-1的hTERT-MSC细胞(克隆体79TM217/13)的悬浮液,3:从结构#217分泌二聚GLP-1的AtT20细胞(克隆体81-A-217/3)的悬浮液,M:预染蛋白质标记[kDa])。该结果显示包含GLP-1(7-37)和C-端附件(图3中的2和3)的本发明肽可从转染细胞系分泌并可使用与GLP-1(7-37)的中部分子的表位结合的抗GLP-1抗体检测到。
实施例4
人类细胞分泌的GLP-1肽的离体稳定性
采用编码异源溶基质素信号序列的结构瞬时转染HEK293和hTERT-MSC细胞,该序列与编码下列肽的GLP-1变异体相连:
1:GLP-1(7-37)
2:GLP-1(7-37)-IP2-延展与11AA
3:GLP1(7-37)-IP2-GLP1(7-37)
将细胞悬浮液(包含从细胞或合成GLP-1(7-37)(巴赫姆)分泌出的GLP-1肽)与人类淋巴细胞浓缩浆(包含二肽基肽酶活性)在37℃下5%CO2中孵化3或4小时。来自空载体转染细胞的悬浮液中的合成GLP-1(7-37)可用作DPP-1V活性的正控制(positive control),该DPP-IV的活性受到加入的DPP-IV抑制剂(#DPP4,生物趋势公司(biotrend))的抑制。使用GLP-1(活性)ELlSA(#EGLP-35K,生物趋势公司),使用与可识别DPP-1V降解的GLP-1(7-37)的N-端相连的抗体(非活性GLP-1(9-37)肽),可以测得活性GLP。
其结果如图4(HEK293细胞)和图5(hTERT-MSC细胞)所示。HEK293细胞和hTERT-MSC细胞均是该基因结构的有效寄主。类型1到3的转染细胞的结果的编号方式与实施例3相同(1:在空载体转染hTERT-MSC细胞的悬浮液中溶解100ng合成GLP-1(7-37),2:从结构#217分泌二聚GLP-1的hTERT-MSC细胞(克隆体79TM217/13)的悬浮液,3:从结构#217分泌二聚GLP-1的AtT20细胞(克隆体81-A-217/3)的悬浮液)。当结构1产生野生型GLP-1(DPP-1V可以与合成GLP-1类似的方式使其失活)时,本发明C-端延伸的GLP-1形式(图4中的2和3,图5中的3)具有更高的抗降解性,并维持至少40%的活性。该C-端延伸的GLP-1肽在离体人类血浆中是相当稳定的。具有二聚GLP-1序列(3)的肽在离体对于DPP-IV降解是完全稳定的。
实施例5
GLP-1肽的蛋白印迹分析
通过固相合成(syn)或使用的E.coli(rec)重组细胞法可合成各种GLP-1肽。将GLP-1肽(31ng SEQ ID No:1,以及SEQ ID No:6、SEQ ID No:7、SEQID No:8各10ng)在10%-20%浓度梯度的SDS PAGE(120V,90分钟)中分散,并通过半干引印迹转移槽(2.0mA/cm2,60分)将其转移到PVDF膜(转印膜0.45μm微孔IPVH 00010)。在甲醇固定和阻断(3%(w∶v)BSA、TBS中的0.1%(v∶v)吐温-20)后,在4℃ o/n使用1μg/ml抗-GLP-1抗体(HYB 147-12,Antibodyshop)免疫印迹该膜。在RT使用0.02μg/ml检测抗体(抗鼠IgG、HRP共轭、珀金埃尔默PC2855-1197)洗涤和孵化4小时后,荧光检测将显示该蛋白质的位置。图6示出了该肽的蛋白印迹分析,给出了下列值:对应于GLP-1(7-37),31aa,3,3kD的SEQ ID No.:1(ID1syn);对应于GLP-1(7-37)-IP2,46aa,5,1kD的SEQ ID No.:8(ID8syn,CM3);对应于GLP-1(7-37)-IP2-RR-GLP2,83aa,9,4kD的SEQ ID No.:7(ID7rec,CM2);对应于GLP-1(7-37)-IP2-RR-GLP1(7-37),79aa,8,7kD的SEQ ID No.:6(ID6syn,CM1)。
实施例6
GLP-1CM肽的离体人类血浆稳定性
将合成GLP-1肽(SEQ ID No:1syn、SEQ ID No:6syn、SEQ ID No:7rec、SEQ IDNo:8syn)与浓度为20ng/mL的人类血浆在37℃下5%CO2中孵化3小时。血浆的二肽基肽酶活性受到DPP-IV抑制剂(#DPP4,生物趋势公司)的抑制。使用GLP-1(活性)ELlSA(#EGLP-35K,生物趋势公司)测量活性GLP。与活性GLP-1(7-37)(SEQ ID No:1)比较,本发明C-端延长的GLP-1肽SEQ ID No:6、SEQID No:7和SEQ ID No:8在离体人类血浆中相当稳定(图7)。作为控制(位于右手边)还示出了在加入DPP-IV的试验中获得的结果。在这些控制试验中,GLP-1活性完全得以维持。
实施例7
离体生物测定
环AMP生成
RIN-5F细胞(鼠胰岛细胞瘤,ECACC No.95090402)在24-孔板上生长4天,达到70%汇聚。在加入含1%HAS(安内特)、02mM IBMX(858455,西格马)和检测肽的0.5mL DMEM(E15-009,PAA)之前,采用DMEM(E15-009,PAA)洗涤细胞两次。在25℃下孵育20分钟后,使用冰冷的PBS冲洗细胞两次。加入含0.5%曲拉通100的0.1N HCl萃取培育的cAMP。使用cAMP(低pH)EIA(Cat.DE0355,R&D)量化环AMP。3*10-8M SEQ ID No:1、SEQ ID No:6syn、SEQ ID No:6rec、SEQ ID No:7rec、SEQ ID No:8syn用于刺激。
其结果如图8所示。100%cAMP产物对应于不含GLP-1的基础产物。GLP-1与连有G蛋白的受体相连并刺激cAMP产物。所有被检测的分子都增加了细胞cAMP产物。
实施例8
在体生物活性
通过一天两次(上午9点和下午5点)皮下注射5μg肽治疗11-周大的患II型糖尿病的老鼠(C57BL/Ks-Leprdb/db,Harlan)(每组n=5)。在用GLPCM肽治疗之前年(0天)或之后(2、4、7、10天),在经过隔夜培养周期后的上午10点,测量血糖。示出了相对于0天时测量的血糖浓度数据。
经测试的所有的本发明肽(SEQ ID No.:6(合成的或重组的)和SEQ IDNo.:7(合成的或重组的))具有抗高血糖功效。重组SEQ ID No.:6(CM1)和合成SEQ ID No.:8(CM3)获得的效果最好。在图9(y-轴)中示出了治疗的相关功效。在day=0时的血糖浓度被设为1。未治疗的动物的血糖水平随着时间持续增加,而采用本发明肽治疗的动物的血糖浓度大体上随着时间而持续降低。
实施例9
GLP1CM肽的离体血浆稳定性测量(动力学测试法)
使用10%人类血浆在37℃下5%CO2中孵化孵育缓冲液(50mM三乙醇胺-HCl(pH 7、8),0.2%HSA)中的1μM肽的等分试样0、3、6和9小时,该肽是来自CM3、谷氨酸取代的CM3类似物(CM3-ANA01、CM3-ANA02、CM3-ANA03、CM3-ANA04)、C-端缩短的CM3类似物(CM3-ANA06、CM3-ANA07)或C-端延长的CM3类似物(CM3-ANA09)。二肽基肽酶活性受到添加的DPP-IV抑制剂(#DPP4,生物趋势公司)的抑制,且使用GLP-1(活性)ELlSA(#EGLP-35K,利高)测量活性GLP。测量结果来自重复三次的至少两个独立试验。
具有至少9个添加到GLP-1的C-端的本发明肽是CM3(小鼠,GLP-1(7-37)-IP2)及其在组分(II)(IP2)具有修饰序列的衍生物,即CM3-ANA01、CM3-ANA01、CM3-ANA02、CM3-ANA03、CM3-ANA04、CM3-ANA05、CM3-ANA07、CM3-ANA07。肽GLP-1和CM3-ANA06反映了控制或参照物。
图10以0小时(0h)值的a%(0h=100%)示出了活性GLP-1的活性部分。可以清楚地看到GLP-1具有最差的血浆稳定性,在暴露在血浆中9小时(9h)后,其稳定性仅为9%。CM3-ANA01展示了最佳的稳定性,在9小时后,还有84%的原料剩余。在9小时以后,本发明肽具有至少30%的剩余物,而具有较短延伸的肽达不到这个值。从动力学,对于受检测物质,可以确定将活性GLP-1降解到0h值的80%(DT80:80%降解时间)所需的时间。
增加DT80值的序列清单:
肽 | 序列 | DT80[h] | 9h值[%] |
GLP-1 | HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG | 0.8 | 9 |
CM3-ANA06 | HAEGTFTSDVSSYLEGQAAKEFLAWLVKGRGRRDFP | 0.9 | 11 |
CM3-ANA03 | HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDAAAAVAIAEELG | 1.6 | 30 |
CM3-ANA07 | HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFPEEVA | 2.0 | 37 |
CM3-ANA09 | HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFPEEVAIAEELGRRHAC | 4.2 | 60 |
CM3 | HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFPEEVAIAEELG | 4.4 | 63 |
CM3-ANA02 | HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFAEEVAIAEELG | 4.5 | 65 |
CM3-ANA04 | HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGAADAAAAVAIAAALG | 4.7 | 65 |
CM3-ANA01 | HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGAADFPEEVAIEELG | 12.5 | 84 |
本发明基质示出了与参照基质相比的长相当多的离体血浆稳定性。不受任何理论的束缚,GLP-1的C-端延展引入了位阻,其阻止类似物进入到DPPIV的活性位点,并且这样避免降解,而较短的参照基质并不展示其对于GLP-1稳定性的保护效果。该假设得到肽CM3-ANA06(36aa)、CM3-ANA07(40aa)和CM3(46aa)的累积C-端延展的持续增加的稳定性支持。然而,该现象并不仅仅取决于氨基酸的纯数量,其已在CM3的IP2区通过谷氨酸取代示出了。CM3-ANA03肽具有与CM3相同的氨基酸数量,其具有与CM3相比大大降低的稳定性。另一方面,CM3-ANA01由46个氨基酸组成,其具有比CM3更高的血浆稳定性。这显示和氨基酸数目比较,附加空间位阻效应可能影响其稳定性。特别优选地是在GLP-1的C-端具有包括IP2的延展部分的肽或是在GLP-1的C-端具有包括与IP2一定程度相似的类似物的延展部分的肽,这是由于其特定的构造,这种构造可阻碍N-端区进入DPPIV的活性位点。
实施例10
载体研究-免疫原性
为了说明测试基质CM1的潜在免疫原性,在Parabioscience(Groningen,德国)完成小鼠研究。该试验在22天内(n=5)接受5次重复注射(70μg肽/剂量,静脉注射)的BALB/c小鼠上完成。
CM1syn没有引入毒性,并仅仅有极微的T-细胞抗体响应和抗体滴度,如下图中所示。不能排出副产品(该肽的纯度仅为95%)的免疫效应。图11示出了对于CM1syn的潜在免疫效应的评估研究。在左手边(图11A),示出了T-细胞回忆响应试验(recall response assay)结果。将经治疗的和校验鼠的脾外植,T细胞分离并采用量逐渐增加的抗原对其进行刺激,测得增生的回忆响应。在右手边(图11B),测定了动物免疫血浆中的抗原特异性IgG抗体滴度。
实施例11
在患糖尿病的鼠中的剂量功效研究
在经过2小时的紧固期后,采用5种不同浓度的GLP-1、CM1、CM3、CM3-ANA01、胰高血糖素样肽-1受体激动剂(exendin-4)对糖尿病C57BL/KsJ@Rj-db(db/db)鼠进行治疗。在注射前、注射后1小时和4小时,分别测定尾巴血液的血糖。
其结果显示如图12中(图12A(GLP-1)、图12B(CM1)、图12C(CM3)、图12D(CM3-ANA01))的剂量-反应曲线所示。对于每个测试基质,采用与不同的浓度初始值相关血糖的百分比差异制作了曲线图。对于不同测试基质的ED50值,可通过使用1h时测得的与基本值相比的百分比的降低值来绘制剂量响应曲线来确定。为了估计GLP-1、CM3-ANA01、CM3rec和exendin-4的ED50值,采用了摩根-默赛尔-弗罗丁(MMF)模型。对于CM1,使用了理查兹(Richards)模型。两个模型都是适合剂量响应估计的S型拟合模型。对于每个肽,可从血糖下降的百分比测量该血糖ED50值(以ug/鼠为单位的单个皮下剂量)。ED50值都在表1中概况出来。
表1:
肽 | ED50[μg/鼠] |
GLP-1(7-37) | 12,76 |
CM1 | 0,61 |
CM3 | 0,25 |
CM3-ANA01 | 0,55 |
exendin-4 | 0,03 |
从ED50值推出,GLP-1类似物CM1、CM3和CM3-ANA01展现了高于20倍的更好的在体生物活性,这很可能是血浆稳定性增强的结果。
至少对于最高浓度,所有的被测试的肽导致高血糖症db/db鼠中的血糖水平的显著降低。表2总结了在1.1-3.0nmol肽(相对校验)的单次皮下注射后血糖的降低。表2示出了被测基质(相对校验)的血糖降低功效。对于注射本发明肽后的时间段1小时(@1h)和4小时(@4h),给出了血糖的降低和对应的重要水平。在这些肽的长期功效中,可观察到不同之处。仅仅只有exendin-4和CM3在4小时后还显示出明显的血糖水平下降。
表2:
实施例12
db/db鼠的长期治疗
研究了n=12的四组C57BL/KsJ@Rj-db(db/db)鼠。
A组采用媒介(0.9%盐),一天治疗一次;
B组采用24nmol/kg测试基质1:CM1rec,一天治疗一次;
C组采用24nmol/kg测试基质2:CM3-ANA01,一天治疗一次;
D组采用24nmol/kg参考基质exendin-4(在本领域已知并得到证明(exendin-4并不是GLP-1的类似物)),一天治疗一次。
在2°°-3°°p.m之间将肽或媒介一天一次(A-D组)地通过皮下注射到鼠背部的皮肤褶襞。该治疗持续18周。从第12周到第18周,对每个组中的12个动物中的六个每天治疗两次。
研究了鼠的各种生理参数,也就是,健康状况(1)、体重(2)、食物消耗(3)、血糖(4)、葡萄糖赖量检测(5)、胰岛素数据(6)、糖化血红蛋白(7)、病理(8)和T细胞的再次刺激(9)。
健康状况(1)在所有的组中均良好,没有见到治疗的副作用。
体重(2)在经过18周的治疗后,所有检测组均有所下降(图13)。在CM3-ANA01组中,相对的体重增加显著降低。图13A示出了盐(A)、CM1(B)、CM3-ANA01(C)或exendin(D)对db/db鼠的体重的治疗效果。每组12个动物的体重平均值在图中标示出了。图13B示出了盐(A)、CM1(B)、CM3-ANA01(C)或exendin(D)对db/db鼠的体重的治疗效果。在治疗122天后的每组12个动物的相对体重增加在图中标示出了。只有采用测试基质CM3-ANA01(组B)的治疗显示了明显较低的体重增加(p<0.001)。
食物消耗(3)与对照值相比,CM1和CM3-ANA01组中的明显较低。图14示出了在122天的治疗时段中,采用盐(A)、CM1(B)、CM3-ANA01(C)或exendin(D)治疗的db/db鼠的周食物消耗。在组B和C中,每只鼠的相对食物消耗显著降低(p<0.001)。
血糖(4)水平已在各种情况下测得。图15A中示出了非禁食血糖水平(在肽注射后的1小时),也就是,使用盐(A)、CM1(B)、CM3-ANA01(C)或exendin(D)在治疗非禁食血糖水平上的功效。在皮下注射24nmol/kg的盐(A)、CM1(B)、CM3-ANA01(C)或exendin(D)1小时后,从尾巴血中测量血糖浓度。与对照值相比,在皮下肽注射1小时后,B组中的非禁食血糖水平下降到55%,C组中的非禁食血糖水平下降到51%,D组中的非禁食血糖水平下降到60%(图15B)。接收治疗的组中,血糖浓度的下降是相当显著的(p<0.0001)。
图15C中示出了禁食血糖水平(在肽注射后的18小时),也就是,使用盐(A)、CM1(B)、CM3-ANA01(C)或exendin(D)在治疗禁食血糖水平上的功效。在皮下注射检测基质18小时后整夜12小时禁食后,从尾巴血中测量血糖浓度。在皮下肽注射18小时后,CM1和CM3-ANA01组中的禁食血糖浓度降低。
腹腔注射葡萄糖耐量检测(5)(IPGTT)在对A-D组的鼠治疗8星期后执行。在检测基质的皮下注射和腹腔注射20%葡萄糖溶液(每kg含1g葡萄糖)后对尾巴血液检测可确定12小时禁食动物的基本血糖值(0分钟)。在葡萄糖注射15、30、45、60、90和120分钟后,进一步检测血糖。图16A示出了所有经治疗的组的葡萄糖耐量的标准化。在图16A中,示出了经盐(A)、CM1(B)、CM3-ANA01(C)或exendin(D)治疗8个星期后,在腹腔注射葡萄糖耐量检测(IPGTT)中的绝对血糖水平(每个组(n=12)的平均值)。图16B中示出了图16A示出的葡萄糖耐量实验数据的另一表现形式,采用起始血糖水平(100%)归一化经盐(A)、CM1(B)、CM3-ANA01(C)或exendin(D)治疗8个星期后在腹腔注射葡萄糖耐量检测(IPGTT)中的相对血糖水平。
胰岛素水平(6)在12小时整夜禁食后对鼠测得。经过18周的治疗,所有经治疗组的鼠与未治疗的对照值相比,明显生成更多的胰岛素。图17示出了在采用0.9%盐(A组)、CM1肽(B组)、CM3-ANA01肽(C组)或exendin(D组)治疗18个星期后的鼠血浆胰岛素水平的数据。在采用血浆样本后,迅速用蛋白酶抑制剂混合物(cocktail)进行处理,以避免胰岛素降解。采用胰岛素鼠超灵敏ELISA(Cat.#EIA-3440,DRG)针对胰岛素分析血清。其意义已由t检验实验确定。
糖化血红蛋白(7)可通过与红细胞(RBC)中的血红蛋白相连的过量血糖形成。因为RBC具有120天的生命周期,糖化血红蛋白的测量值给出了血糖控制的长期指示,并可将其看作是比血糖水平更好的指标。从球后窦中采集全血,并采用酶HbA1c实验盒(#DZ121A,Diazyme)进行分析,以确定糖化血红蛋白水平。将糖化血红蛋白作为参量,可在所有的经治疗的组中的看到其增加有显著的降低。
图18示出了在采用盐(A)、CM1(B)、CM3-ANA01(C)或exendin(D)治疗6、8、12个星期后,糖化血红蛋白(GHb)在全部血液样本中的相对增加。图18B示出了采用盐(A)、CM1(B)、CM3-ANA01(C)或exendin(D)治疗18个星期后,糖化血红蛋白(GHb)在全部血液样本中的相对增加。给出了每组所有动物的平均值(n=12)。所有的经治疗的组的糖化血红蛋白水平的增加明显低于参照值。
被治疗的鼠的病理(8)也得到研究。验尸结果显示,与没有治疗的组A相比,除胰腺体积外,经治疗的组的器官没有任何变化。在所有的经治疗的组中,测定的胰腺的重量相当地高。图19示出了在采用0.9%盐(A组,n=11)、CM1肽(B组,n=12)、CM3-ANA01肽(C组,n=12)或exendin(D组,n=12)治疗18个星期后,,该动物被屠宰的当天的胰腺重量。
为了评估检测基质的可能免疫功效,通过T细胞再次刺激法来检验IV型 免疫性质(9)。因此将每组8个动物的脾外植,进行T细胞分离并采用不同浓度对应实验基质对其进行再次刺激。与未经治疗的参照值相比,肽再次刺激的T细胞增生没有显著的增加。图20示出了脾细胞对不同浓度的实验基质(CM1(图20A)、CM3-ANA01(图20B)和参考基质exendin-4(图20C))的离体回忆响应。使用非放射性细胞增植实验(细胞增植ELISA、BrdU,化学发光的),三次测量A组到D组的单鼠的脾细胞对与始于50μg/ml的检测基质CM1、CM3-ANA01、参考基质exendin-4的三倍稀释剂的离体回忆响应。可将刺激指数(SI)计算为出现肽情况下的响应和肽缺失的情况下的响应的比值。图中示出了各个治疗组和参照组的平均值±标准差(A组:n=3,B组:n=6,C组:n=7,D组:n=6)。对于均值的计算,仅对具有5μg/ml的Con A的正控制培养物中SI大于6.5的鼠进行了分析。
对患糖尿病的鼠的长期研究进一步支持本发明C-端延长肽的功效。在所有的检测参数中(体重、食物消耗、血糖水平、糖化血红蛋白、葡萄糖耐性、胰岛素分泌、胰腺重量),C-端延长的肽示出了显著的治疗效果。考虑到具有内生天然序列的C-端延长的细胞医学肽的同源性,已经表明与非哺乳动物exendin-4相比,本发明肽在免疫原性方面更具有优势。对鼠的免疫原性研究的数据支持任何临床相关的CM1肽的免疫原性。
序列表
HAEGTFTSDV SSYLEGQAAK EFIAWLVKGR G
Sequence ID No.:1
RRDFPEEVAI VEELG
Sequence ID No.:2
RRDFPEEVAI AEELG
Sequence ID No.:3
HADGSFSDEM NTILDNLAAR DFINWLIQTK ITDRK
Sequence ID No.:4
HADGSFSDEM STILDNLATR DFINWLIQTK ITDKK
Sequence ID No.:5
HAEGTFTSDV SSYLEGQAAK EFIAWLVKGR GRRDFPEEVA IAEELGRRHA
EGTFTSDVSS YLEGQAAKEF IAWLVKGRG
Sequence ID No.:6
HAEGTFTSDV SSYLEGQAAK EFIAWLVKGR GRRDFPEEVA IAEELGRRHA
DGSFSDEMST ILDNLATRDF INWLIQTKIT DKK
Sequence ID No.:7
HAEGTFTSDV SSYLEGQAAK EFIAWLVKGR GRRDFPEEVA IAEELG
Sequence ID No.:8
MAPAAWLRSA AARALLPPML LLLLQPPPLL ARALPPDVHH LHAERRGPQP
WHAALPSSPA PAPATQEAPR PASSLRPPRC GVPDPSDGLS ARNRQKR
Sequence ID No.:9
HAEGTFTSDV SSYLEGQAAK EFIAWLVKGR GRRDFPEEVA IVEELGRRHA
EGTFTSDVSS YLEGQAAKEF IAWLVKGRG
Sequence ID No.:10
HAEGTFTSDV SSYLEGQAAK EFIAWLVKGR GRRDFPEEVA IVEELGRRHA
DGSFSDEMNT ILDNLAARDF INWLIQTKIT DRK
Sequence ID No.:11
HAEGTFTSDV SSYLEGQAAK EFIAWLVKGR GRRDFPEEVA IVEELG
Sequence ID No.:12
1 gatatccacc atggcccccg ccgcctggct gaggagcgcc gccgccaggg
M A P A A W L R S A A A R A
51 ccctgctgcc acccatgctg ctgctgctgc tgcagccccc acctctgctg
L L P P M L L L L L Q P P P L L
101 gcccgggccc tgcccccggt gagtgcccgc cactcgccgt ccgctcctcg
A R A L P P
151 ctgagggggc gccgggcacg cgggctgggc ccagcggcgt atccggacgc
201 caagaaacca gagagccagc cagatgccaa agggccctgc catgtgccgg
251 tgccctttcc ctctccattt gccctgccac acagtgggct ggggttgcac
301 gtgtgtttgc tgacaggcca catctctaac tgtgggccat gtggacctta
351 ggcctgacca gaccctcatg tcttcctcct tcccaggacg tgcaccacct
D V H H L
401 gcacgccgag aggcgcggcc ctcagccctg gcacgccgcc ctgccaagca
H A E R R G P Q P W H A A L P S S
451 gccctgcccc tgccccagcc acccaggagg cccccaggcc tgccagcagc
P A P A P A T Q E A P R P A S S
501 ctgaggccac ccaggtgcgg cgtgcctgat ccctccgatg gcctgagcgc
L R P P R C G V P D P S D G L S A
551 tcggaatcgg cagaagaggc acgccgaggg caccttcacc tccgacgtga
R N R Q K R H A E G T F T S D V S
601 gcagctacct ggagggccag gccgccaagg agttcatcgc ctggctggtg
S Y L E G Q A A K E F I A W L V
651 aagggcaggg gccgcaggga cttccctgag gaggtggcca tcgtggagga
K G R G R R D F P E E V A I V E E
701 gctgggccgg cgacacgccg agggcacctt cacctccgac gtgagcagct
L G R R H A E G T F T S D V S S Y
751 acctggaggg ccaggccgcc aaggagttca tcgcctggct ggtgaagggc
L E G Q A A K E F I A W L V K G
801 aggggctgag cgcgc
R G *
Sequence ID No.:13
1 gatatccacc atggcccccg ccgcctggct gaggagcgcc gccgccaggg ccctgctgcc
61 acccatgctg ctgctgctgc tgcagccccc acctctgctg gcccgggccc tgcccccggt
121 gagtgcccgc cactcgccgt ccgctcctcg ctgagggggc gccgggcacg cgggctgggc
181 ccagcggcgt atccggacgc caagaaacca gagagccagc cagatgccaa agggccctgc
241 catgtgccgg tgccctttcc ctctccattt gccctgccac acagtgggct ggggttgcac
301 gtgtgtttgc tgacaggcca catctctaac tgtgggccat gtggacctta ggcctgacca
361 gaccctcatg tcttcctcct tcccaggacg tgcaccacct gcacgccgag aggcgcggcc
421 ctcagccctg gcacgccgcc ctgccaagca gccctgcccc tgccccagcc acccaggagg
481 cccccaggcc tgccagcagc ctgaggccac ccaggtgcgg cgtgcctgat ccctccgatg
541 gcctgagcgc tcggaatcgg cagaagaggc acgccgaggg caccttcacc tccgacgtga
601 gcagctacct ggagggccag gccgccaagg agttcatcgc ctggctggtg aagggcaggg
661 gccgcaggga cttccctgag gaggtggcca tcgtggagga gctgggccgg cgacacgccg
721 agggcacctt cacctccgac gtgagcagct acctggaggg ccaggccgcc aaggagttca
781 tcgcctggct ggtgaagggc aggggctgag cgcgc
Sequence ID No.:14
1 gatatccacc atggcccccg ccgcctggct gaggagcgcc gccgccaggg
M A P A A W L R S A A A R A
51 ccctgctgcc acccatgctg ctgctgctgc tgcagccccc acctctgctg
L L P P M L L L L L Q P P P L L
101 gcccgggccc tgcccccggt gagtgcccgc cactcgccgt ccgctcctcg
A R A L P P
151 ctgagggggc gccgggcacg cgggctgggc ccagcggcgt atccggacgc
201 caagaaacca gagagccagc cagatgccaa agggccctgc catgtgccgg
251 tgccctttcc ctctccattt gccctgccac acagtgggct ggggttgcac
301 gtgtgtttgc tgacaggcca catctctaac tgtgggccat gtggacctta
351 ggcctgacca gaccctcatg tcttcctcct tcccaggacg tgcaccacct
D V H H L
401 gcacgccgag aggcgcggcc ctcagccctg gcacgccgcc ctgccaagca
H A E R R G P Q P W H A A L P S S
451 gccctgcccc tgccccagcc acccaggagg cccccaggcc tgccagcagc
P A P A P A T Q E A P R P A S S
501 ctgaggccac ccaggtgcgg cgtgcctgat ccctccgatg gcctgagcgc
L R P P R C G V P D P S D G L S A
551 tcggaatcgg cagaagaggc acgccgaggg caccttcacc tccgacgtga
R N R Q K R H A E G T F T S D V S
601 gcagctacct ggagggccag gccgccaagg agttcatcgc ctggctggtg
S Y L E G Q A A K E F I A W L V
651 aagggcaggg gccgcaggga cttccctgag gaggtggcca tcgtggagga
K G R G R R D F P E E V A I V E E
701 gctgggccgg cgacacgccg acggcagctt cagcgacgag atgaacacca
L G R R H A D G S F S D E M N T I
751 tcctggacaa cctggccgcg cgcgacttca tcaactggct gatccagacc
L D N L A A R D F I N W L I Q T
801 aagatcaccg atcggaagtg agcgcgctga tatc
K I T D R K *
Sequence ID No.:15
1 gatatccacc atggcccccg ccgcctggct gaggagcgcc gccgccaggg ccctgctgcc
61 acccatgctg ctgctgctgc tgcagccccc acctctgctg gcccgggccc tgcccccggt
121 gagtgcccgc cactcgccgt ccgctcctcg ctgagggggc gccgggcacg cgggctgggc
181 ccagcggcgt atccggacgc caagaaacca gagagccagc cagatgccaa agggccctgc
241 catgtgccgg tgccctttcc ctctccattt gccctgccac acagtgggct ggggttgcac
301 gtgtgtttgc tgacaggcca catctctaac tgtgggccat gtggacctta ggcctgacca
361 gaccctcatg tcttcctcct tcccaggacg tgcaccacct gcacgccgag aggcgcggcc
421 ctcagccctg gcacgccgcc ctgccaagca gccctgcccc tgccccagcc acccaggagg
481 cccccaggcc tgccagcagc ctgaggccac ccaggtgcgg cgtgcctgat ccctccgatg
541 gcctgagcgc tcggaatcgg cagaagaggc acgccgaggg caccttcacc tccgacgtga
601 gcagctacct ggagggccag gccgccaagg agttcatcgc ctggctggtg aagggcaggg
661 gccgcaggga cttccctgag gaggtggcca tcgtggagga gctgggccgg cgacacgccg
721 acggcagctt cagcgacgag atgaacacca tcctggacaa cctggccgcg cgcgacttca
781 tcaactggct gatccagacc aagatcaccg atcggaagtg agcgcgctga tatc
Sequence ID No.:16
1 gatatccacc atggcccccg ccgcctggct gaggagcgcc gccgccaggg
M A P A A W L R S A A A R A
51 ccctgctgcc acccatgctg ctgctgctgc tgcagccccc acctctgctg
L L P P M L L L L L Q P P P L L
101 gcccgggccc tgcccccggt gagtgcccgc cactcgccgt ccgctcctcg
A R A L P P
151 ctgagggggc gccgggcacg cgggctgggc ccagcggcgt atccggacgc
201 caagaaacca gagagccagc cagatgccaa agggccctgc catgtgccgg
251 tgccctttcc ctctccattt gccctgccac acagtgggct ggggttgcac
301 gtgtgtttgc tgacaggcca catctctaac tgtgggccat gtggacctta
351 ggcctgacca gaccctcatg tcttcctcct tcccaggacg tgcaccacct
D V H H L
401 gcacgccgag aggcgcggcc ctcagccctg gcacgccgcc ctgccaagca
H A E R R G P Q P W H A A L P S S
451 gccctgcccc tgccccagcc acccaggagg cccccaggcc tgccagcagc
P A P A P A T Q E A P R P A S S
501 ctgaggccac ccaggtgcgg cgtgcctgat ccctccgatg gcctgagcgc
L R P P R C G V P D P S D G L S A
551 tcggaatcgg cagaagaggc acgccgaggg caccttcacc tccgacgtga
R N R Q K R H A E G T F T S D V S
601 gcagctacct ggagggccag gccgccaagg agttcatcgc ctggctggtg
S Y L E G Q A A K E F I A W L V
651 aagggcaggg gccgcaggga cttccctgag gaggtggcca tcgtggagga
K G R G R R D F P E E V A I V E E
701 gctgggccgg cgacacgccg acggcagctt cagcgacgag atgaacacca
L G R R H A D G S F S D E M N T I
751 tcctggacaa cctggccgcg cgctga tat c
L D N L A A R *
Sequence ID No.:17
1 gatatccacc atggcccccg ccgcctggct gaggagcgcc gccgccaggg ccctgctgcc
61 acccatgctg ctgctgctgc tgcagccccc acctctgctg gcccgggccc tgcccccggt
121 gagtgcccgc cactcgccgt ccgctcctcg ctgagggggc gccgggcacg cgggctgggc
181 ccagcggcgt atccggacgc caagaaacca gagagccagc cagatgccaa agggccctgc
241 catgtgccgg tgccctttcc ctctccattt gccctgccac acagtgggct ggggttgcac
301 gtgtgtttgc tgacaggcca catctctaac tgtgggccat gtggacctta ggcctgacca
361 gaccctcatg tcttcctcct tcccaggacg tgcaccacct gcacgccgag aggcgcggcc
421 ctcagccctg gcacgccgcc ctgccaagca gccctgcccc tgccccagcc acccaggagg
481 cccccaggcc tgccagcagc ctgaggccac ccaggtgcgg cgtgcctgat ccctccgatg
541 gcctgagcgc tcggaatcgg cagaagaggc acgccgaggg caccttcacc tccgacgtga
601 gcagctacct ggagggccag gccgccaagg agttcatcgc ctggctggtg aagggcaggg
721 acggcagctt cagcgacgag atgaacacca tcctggacaa cctggccgcg cgctgatatc
Sequence ID No.:18
1 gatatccacc atggcccccg ccgcctggct gaggagcgcc gccgccaggg
M A P A A W L R S A A A R A
51 ccctgctgcc acccatgctg ctgctgctgc tgcagccccc acctctgctg
L L P P M L L L L L Q P P P L L
101 gcccgggccc tgcccccggt gagtgcccgc cactcgccgt ccgctcctcg
A R A L P P
151 ctgagggggc gccgggcacg cgggctgggc ccagcggcgt atccggacgc
201 caagaaacca gagagccagc cagatgccaa agggccctgc catgtgccgg
251 tgccctttcc ctctccattt gccctgccac acagtgggct ggggttgcac
301 gtgtgtttgc tgacaggcca catctctaac tgtgggccat gtggacctta
351 ggcctgacca gaccctcatg tcttcctcct tcccaggacg tgcaccacct
D V H H L
401 gcacgccgag aggcgcggcc ctcagccctg gcacgccgcc ctgccaagca
H A E R R G P Q P W H A A L P S S
451 gccctgcccc tgccccagcc acccaggagg cccccaggcc tgccagcagc
P A P A P A T Q E A P R P A S S
501 ctgaggccac ccaggtgcgg cgtgcctgat ccctccgatg gcctgagcgc
L R P P R C G V P D P S D G L S A
551 tcggaatcgg cagaagaggc acgccgaggg caccttcacc tccgacgtga
R N R Q K R H A E G T F T S D V S
601 gcagctacct ggagggccag gccgccaagg agttcatcgc ctggctggtg
S Y L E G Q A A K E F I A W L V
651 aagggcaggg gccgcaggga cttccctgag gaggtggcca tcgtggagga
K G R G R R D F P E E V A I V E E
701 gctgggctga gcgcgc
L G *
Sequence ID No.:19
1 gatatccacc atggcccccg ccgcctggct gaggagcgcc gccgccaggg ccctgctgcc
61 acccatgctg ctgctgctgc tgcagccccc acctctgctg gcccgggccc tgcccccggt
121 gagtgcccgc cactcgccgt ccgctcctcg ctgagggggc gccgggcacg cgggctgggc
181 ccagcggcgt atccggacgc caagaaacca gagagccagc cagatgccaa agggccctgc
241 catgtgccgg tgccctttcc ctctccattt gccctgccac acagtgggct ggggttgcac
301 gtgtgtttgc tgacaggcca catctctaac tgtgggccat gtggacctta ggcctgacca
361 gaccctcatg tcttcctcct tcccaggacg tgcaccacct gcacgccgag aggcgcggcc
421 ctcagccctg gcacgccgcc ctgccaagca gccctgcccc tgccccagcc acccaggagg
481 cccccaggcc tgccagcagc ctgaggccac ccaggtgcgg cgtgcctgat ccctccgatg
541 gcctgagcgc tcggaatcgg cagaagaggc acgccgaggg caccttcacc tccgacgtga
601 gcagctacct ggagggccag gccgccaagg agttcatcgc ctggctggtg aagggcaggg
661 gccgcaggga cttccctgag gaggtggcca tcgtggagga gctgggctga gcgcgc
Sequence ID No.:20
HGEGTFTSDV SSYLEGQAAK EFIAWLVKGR G
Sequence ID No.:21
RRDFPEEVAI
Sequence ID No.:22
RRDFPEEVAI VEEL
Sequence ID No.:23
RRDFPEEVAI AEEL
Sequence ID No.:24
Asp Phe Pro Glu Glu Val Ala
Sequence ID No:25
Ala Ala Asp Phe Pro Glu Glu Val Ala Ile
Sequence ID No:26
Ala Ala Asp Phe Pro Glu Glu Val Ala Ile Val Glu Glu Leu
Sequence ID No:27
Ala Ala Asp Phe Pro G1u Glu Val Ala Ile Ala Glu Glu Leu
Sequence ID No:28
Ala Ala Asp Phe Pro Glu Glu Val Ala Ile Val Glu Glu Leu Gly
Sequence ID No:29
Ala Ala Asp Phe Pro Glu Glu Val Ala Ile Ala Glu Glu Leu Gly
Sequence ID No:30
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Ala
Ala Asp Phe Pro Glu Glu Val Ala Ile Ala Glu Glu Leu Gly
Sequence ID No:31
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
Arg Asp Phe Ala Glu Glu Val Ala Ile Ala Glu Glu Leu Gly
Sequence ID No:32
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu ValLys Gly Arg Gly Arg
Arg Asp Ala Ala Ala Ala Val Ala Ile Ala Glu Glu Leu Gly
Sequence ID No:33
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Ala
Ala Asp Ala Ala Ala Ala Val Ala Ile Ala Ala Ala Leu Gly
Sequence ID No:34
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
Arg Asp Phe Pro
Sequence ID No:35
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
Arg Asp Phe Pro Glu Glu Val Ala
Sequence ID No:36
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
Arg Asp Phe Pro Glu Glu Val Ala Ile Ala Glu Glu Leu Gly Arg Arg
His Ala Cys
Sequence ID No:37
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Ala
Ala Asp Phe Pro Glu Glu Val Ala Ile Val Glu Glu Leu Gly
Sequence ID No:38
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
Arg Asp Phe Ala Glu Glu Val Ala Ile Val Glu Glu Leu Gly
Sequence ID No:39
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
Arg Asp Ala Ala Ala Ala Val Ala Ile Val Glu Glu Leu Gly
Sequence ID No:40
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Ala
Ala Asp Ala Ala Ala Ala Val Ala Ile Val Ala Ala Leu Gly
Sequence ID No:41
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
Arg Asp Phe Pro Glu Glu Val Ala Ile Val Glu Glu Leu Gly Arg Arg
His Ala Cys
Sequence ID No:42
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Xaa
Sequence ID No:43(分子式(I))
Xaa Xaa Glu Gly Thr Phe Thr Ser Asp Xaa Ser Xaa Xaa Xaa Glu Xaa
Xaa Ala Xaa Xaa Xaa Phe Ile Xaa Trp Leu Xaa Xaa Xaa Xaa Xaa
Sequence ID No:44(分子式(I))
Xaa Xaa Glu Gly Thr Phe Thr Ser Asp Val Ser Xaa Tyr Leu Glu Xaa
Xaa Ala Ala Xaa Glu Phe Ile Xaa Trp Leu Val Xaa Xaa Xaa Xaa
Sequence ID No:45(分子式(III))
序列表
<110>生物相容英国公司
<120>增强肽酶抗性的GLP-1(胰高血糖素样肽-1)融合多肽
<130>WCN0810058CXH-BGSJ
<140>
<141>
<150>EP050207.18.2
<151>2005-09-22
<160>45
<170>PatentIn version 3.3
<210>1
<211>31
<212>PRT
<213>人工序列
<220>
<223>序列描述:对应GLP-1(7-37)的合成肽(参见说明书第36页)
<400>1
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly
20 25 30
<210>2
<211>15
<212>PRT
<213>人工序列
<220>
<223>序列描述:全长IP-2序列,具有天然存在的IP-2序列的所有15个氨基酸,人类的
(参见说明书第36页)
<400>2
Arg Arg Asp Phe Pro Glu Glu Val Ala Ile Val Glu Glu Leu Gly
1 5 10 15
<210>3
<211>15
<212>PRT
<213>人工序列
<220>
<223>序列描述:全长IP-2序列,具有天然存在的IP-2序列的所有15个氨基酸,鼠
(参见说明书第36页)
<400>3
Arg Arg Asp Phe Pro Glu Glu Val Ala Ile Ala Glu Glu Leu Gly
1 5 10 15
<210>4
<211>35
<212>PRT
<213>人工序列
<220>
<223>
序列描述:GLP-2的鼠异构体形式(参见说明书第36页)
<400>4
His Ala Asp Gly Ser Phe Ser Asp Glu Met Asn Thr Ile Leu Asp Asn
1 5 10 15
Leu Ala Ala Arg Asp Phe Ile Asn Trp Leu Ile Gln Thr Lys Ile Thr
20 25 30
Asp Arg Lys
35
<210>5
<211>35
<212>PRT
<213>人工序列
<220>
<223>
序列描述:GLP-2的人异构体形式(参见说明书第36页)
<400>5
His Ala Asp Gly Ser Phe Ser Asp Glu Met Ser Thr Ile Leu Asp Asn
1 5 10 15
Leu Ala Thr Arg Asp Phe Ile Asn Trp Leu Ile Gln Thr Lys Ile Thr
20 25 30
Asp Lys Lys
35
<210>6
<211>79
<212>PRT
<213>人工序列
<220>
<223>序列描述:SEQ ID No:6(ID6syn,CM1)对应
GLP-1(7-37)-IP2-RR-GLP1(7-37)、79aa、8、7kD(参见说明书第36页)
<400>6
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
20 25 30
Arg Asp Phe Pro Glu Glu Val Ala Ile Ala Glu Glu Leu Gly Arg Arg
35 40 45
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
50 55 60
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly
65 70 75
<210>7
<211>83
<212>PRT
<213>人工序列
<220>
<223>序列描述:SEQ ID No:7(ID7rec,CM2)对应
GLP-1(7-37)-IP2-RR-GLP2、83aa、9、4kD(参见说明书第36页)
<400>7
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
20 25 30
Arg Asp Phe Pro Glu Glu Val Ala Ile Ala Glu Glu Leu Gly Arg Arg
35 40 45
His Ala Asp Gly Ser Phe Ser Asp Glu Met Ser Thr Ile Leu Asp Asn
50 55 60
Leu Ala Thr Arg Asp Phe Ile Asn Trp Leu Ile Gln Thr Lys Ile Thr
65 70 75 80
Asp Lys Lys
<210>8
<211>46
<212>PRT
<213>人工序列
<220>
<223>序列描述:SEQ ID No:8(ID8 syn,CM3)对应
GLP-1(7-37)-IP2、46aa、5、1kD;
<400>8
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
20 25 30
Arg Asp Phe Pro Glu Glu Val Ala Ile Ala Glu Glu Leu Gly
35 40 45
<210>9
<211>97
<212>PRT
<213>人工序列
<220>
<223>序列描述:人工序列
<400>9
Met Ala Pro Ala Ala Trp Leu Arg Ser Ala Ala Ala Arg Ala Leu Leu
1 5 10 15
Pro Pro Met Leu Leu Leu Leu Leu Gln Pro Pro Pro Leu Leu Ala Arg
20 25 30
Ala Leu Pro Pro Asp Val His His Leu His Ala Glu Arg Arg Gly Pro
35 40 45
Gln Pro Trp His Ala Ala Leu Pro Ser Ser Pro Ala Pro Ala Pro Ala
50 55 60
Thr Gln Glu Ala Pro Arg Pro Ala Ser Ser Leu Arg Pro Pro Arg Cys
65 70 75 80
Gly Val Pro Asp Pro Ser Asp Gly Leu Ser Ala Arg Asn Arg Gln Lys
85 90 95
Arg
<210>10
<211>79
<212>PRT
<213>人工序列
<220>
<223>序列描述:SEQ ID No:10
(N-GLP-1(7-37)-IP2(人)-RR-GLP-1(7-37)-C,同样指定人CM1),(参见说明书第36页)
<400>10
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
20 25 30
Arg Asp Phe Pro Glu Glu Val Ala Ile Val Glu Glu Leu Gly Arg Arg
35 40 45
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
50 55 60
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly
65 70 75
<210>11
<211>83
<212>PRT
<213>人工序列
<220>
<223>序列描述:SEQ ID No:11
(N-GLP-1(7-37)-IP2(人)-RR-GLP-2-C),在此同样指定人CM2),(参见说明书第36页)
<400>11
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
20 25 30
Arg Asp Phe Pro Glu Glu Val Ala Ile Val Glu Glu Leu Gly Arg Arg
35 40 45
His Ala Asp Gly Ser Phe Ser Asp Glu Met Asn Thr Ile Leu Asp Asn
50 55 60
Leu Ala Ala Arg Asp Phe Ile Asn Trp Leu Ile Gln Thr Lys Ile Thr
65 70 75 80
Asp Arg Lys
<210>12
<211>46
<212>PRT
<213>人工序列
<220>
<223>序列描述:SEQ ID No:12,GLP-1(7-37)不通过任何接头直接通过其C端与
人IP2连接(参见说明书第36页)
<400>12
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
20 25 30
Arg Asp Phe Pro Glu Glu Val Ala Ile Val Glu Glu Leu Gly
35 40 45
<210>13
<211>815
<212>DNA
<213>人工序列
<220>
<223>序列描述:(SEQ ID No:13)展现依照图1K的结构的转染肽序列(SEQ ID No:14),
(参见说明书第37页);
<220>
<221>CDS
<222>(11)..(118)
<220>
<221>CDS
<222>(387)..(809)
<400>13
gatatccacc atg gcc ccc gcc gcc tgg ctg agg agc gcc gcc gcc agg 49
Met Ala Pro Ala Ala Trp Leu Arg Ser Ala Ala Ala Arg
1 5 10
gcc ctg ctg cca ccc atg ctg ctg ctg ctg ctg cag ccc cca cct ctg 97
Ala Leu Leu Pro Pro Met Leu Leu Leu Leu Leu Gln Pro Pro Pro Leu
15 20 25
ctg gcc cgg gcc ctg ccc ccg gtgagtgccc gccactcgcc gtccgctcct 148
Leu Ala Arg Ala Leu Pro Pro
30 35
cgctgagggg gcgccgggca cgcgggctgg gcccagcggc gtatccggac gccaagaaac 208
cagagagcca gccagatgcc aaagggccct gccatgtgcc ggtgcccttt ccctctccat 268
ttgccctgcc acacagtggg ctggggttgc acgtgtgttt gctgacaggc cacatctcta 328
actgtgggcc atgtggacct taggcctgac cagaccctca tgtcttcctc cttcccag 386
gac gtg cac cac ctg cac gcc gag agg cgc ggc cct cag ccc tgg cac 434
Asp Val His His Leu His Ala Glu Arg Arg Gly Pro Gln Pro Trp His
40 45 50
gcc gcc ctg cca agc agc cct gcc cct gcc cca gcc acc cag gag gcc 482
Ala Ala Leu Pro Ser Ser Pro Ala Pro Ala Pro Ala Thr Gln Glu Ala
55 60 65
ccc agg cct gcc agc agc ctg agg cca ccc agg tgc ggc gtg cct gat 530
Pro Arg Pro Ala Ser Ser Leu Arg Pro Pro Arg Cys Gly Val Pro Asp
70 75 80
ccc tcc gat ggc ctg agc gct cgg aat cgg cag aag agg cac gcc gag 578
Pro Ser Asp Gly Leu Ser Ala Arg Asn Arg Gln Lys Arg His Ala Glu
85 90 95 100
ggc acc ttc acc tcc gac gtg agc agc tac ctg gag ggc cag gcc gcc 626
Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly Gln Ala Ala
105 110 115
aag gag ttc atc gcc tgg ctg gtg aag ggc agg ggc cgc agg gac ttc 674
Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg Arg Asp Phe
120 125 130
cct gag gag gtg gcc atc gtg gag gag ctg ggc cgg cga cac gcc gag 722
Pro Glu Glu Val Ala Ile Val Glu Glu Leu Gly Arg Arg His Ala Glu
135 140 145
ggc acc ttc acc tcc gac gtg agc agc tac ctg gag ggc cag gcc gcc 770
Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly Gln Ala Ala
150 155 160
aag gag ttc atc gcc tgg ctg gtg aag ggc agg ggc tga gcgcgc 815
Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly
165 170 175
<210>14
<211>815
<212>DNA
<213>人工序列
<220>
<223>序列描述:依照图1K的结构的DNA序列(参见说明书第37页);
<400>14
gatatccacc atggcccccg ccgcctggct gaggagcgcc gccgccaggg ccctgctgcc 60
acccatgctg ctgctgctgc tgcagccccc acctctgctg gcccgggccc tgcccccggt 120
gagtgcccgc cactcgccgt ccgctcctcg ctgagggggc gccgggcacg cgggctgggc 180
ccagcggcgt atccggacgc caagaaacca gagagccagc cagatgccaa agggccctgc 240
catgtgccgg tgccctttcc ctctccattt gccctgccac acagtgggct ggggttgcac 300
gtgtgtttgc tgacaggcca catctctaac tgtgggccat gtggacctta ggcctgacca 360
gaccctcatg tcttcctcct tcccaggacg tgcaccacct gcacgccgag aggcgcggcc 420
ctcagccctg gcacgccgcc ctgccaagca gccctgcccc tgccccagcc acccaggagg 480
cccccaggcc tgccagcagc ctgaggccac ccaggtgcgg cgtgcctgat ccctccgatg 540
gcctgagcgc tcggaatcgg cagaagaggc acgccgaggg caccttcacc tccgacgtga 600
gcagctacct ggagggccag gccgccaagg agttcatcgc ctggctggtg aagggcaggg 660
gccgcaggga cttccctgag gaggtggcca tcgtggagga gctgggccgg cgacacgccg 720
agggcacctt cacctccgac gtgagcagct acctggaggg ccaggccgcc aaggagttca 780
tcgcctggct ggtgaagggc aggggctgag cgcgc 815
<210>15
<211>834
<212>DNA
<213>人工序列
<220>
<223>序列描述:依照图1h的转译肽的序列和DNA序列(参见说明书第38页);
<220>
<221>CDS
<222>(11)..(118)
<220>
<221>CDS
<222>(387)..(821)
<400>15
gatatccacc atg gcc ccc gcc gcc tgg ctg agg agc gcc gcc gcc agg 49
Met Ala Pro Ala Ala Trp Leu Arg Ser Ala Ala Ala Arg
1 5 10
gcc ctg ctg cca ccc atg ctg ctg ctg ctg ctg cag ccc cca cct ctg 97
Ala Leu Leu Pro Pro Met Leu Leu Leu Leu Leu Gln Pro Pro Pro Leu
15 20 25
ctg gcc cgg gcc ctg ccc ccg gtgagtgccc gccactcgcc gtccgctcct 148
Leu Ala Arg Ala Leu Pro Pro
30 35
cgctgagggg gcgccgggca cgcgggctgg gcccagcggc gtatccggac gccaagaaac 208
cagagagcca gccagatgcc aaagggccct gccatgtgcc ggtgcccttt ccctctccat 268
ttgccctgcc acacagtggg ctggggttgc acgtgtgttt gctgacaggc cacatctcta 328
actgtgggcc atgtggacct taggcctgac cagaccctca tgtcttcctc cttcccag 386
gac gtg cac cac ctg cac gcc gag agg cgc ggc cct cag ccc tgg cac 434
Asp Val His His Leu His Ala Glu Arg Arg Gly Pro Gln Pro Trp His
40 45 50
gcc gcc ctg cca agc agc cct gcc cct gcc cca gcc acc cag gag gcc 482
Ala Ala Leu Pro Ser Ser Pro Ala Pro Ala Pro Ala Thr Gln Glu Ala
55 60 65
ccc agg cct gcc agc agc ctg agg cca ccc agg tgc ggc gtg cct gat 530
Pro Arg Pro Ala Ser Ser Leu Arg Pro Pro Arg Cys Gly Val Pro Asp
70 75 80
ccc tcc gat ggc ctg agc gct cgg aat cgg cag aag agg cac gcc gag 578
Pro Ser Asp Gly Leu Ser Ala Arg Asn Arg Gln Lys Arg His Ala Glu
85 90 95 100
ggc acc ttc acc tcc gac gtg agc agc tac ctg gag ggc cag gcc gcc 626
Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly Gln Ala Ala
105 110 115
aag gag ttc atc gcc tgg ctg gtg aag ggc agg ggc cgc agg gac ttc 674
Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg Arg Asp Phe
120 125 130
cct gag gag gtg gcc atc gtg gag gag ctg ggc cgg cga cac gcc gac 722
Pro Glu Glu Val Ala Ile Val Glu Glu Leu Gly Arg Arg His Ala Asp
135 140 145
ggc agc ttc agc gac gag atg aac acc atc ctg gac aac ctg gcc gcg 770
Gly Ser Phe Ser Asp Glu Met Asn Thr Ile Leu Asp Asn Leu Ala Ala
150 155 160
cgc gac ttc atc aac tgg ctg atc cag acc aag atc acc gat cgg aag 818
Arg Asp Phe Ile Asn Trp Leu Ile Gln Thr Lys Ile Thr Asp Arg Lys
165 170 175 180
tga gcgcgctgat atc 834
<210>16
<211>834
<212>DNA
<213>人工序列
<220>
<223>序列描述:依照图1h的结构的DNA序列(参见说明书第38页);
<400>16
gatatccacc atggcccccg ccgcctggct gaggagcgcc gccgccaggg ccctgctgcc 60
acccatgctg ctgctgctgc tgcagccccc acctctgctg gcccgggccc tgcccccggt 120
gagtgcccgc cactcgccgt ccgctcctcg ctgagggggc gccgggcacg cgggctgggc 180
ccagcggcgt atccggacgc caagaaacca gagagccagc cagatgccaa agggccctgc 240
catgtgccgg tgccctttcc ctctccattt gccctgccac acagtgggct ggggttgcac 300
gtgtgtttgc tgacaggcca catctctaac tgtgggccat gtggacctta ggcctgacca 360
gaccctcatg tcttcctcct tcccaggacg tgcaccacct gcacgccgag aggcgcggcc 420
ctcagccctg gcacgccgcc ctgccaagca gccctgcccc tgccccagcc acccaggagg 480
cccccaggcc tgccagcagc ctgaggccac ccaggtgcgg cgtgcctgat ccctccgatg 540
gcctgagcgc tcggaatcgg cagaagaggc acgccgaggg caccttcacc tccgacgtga 600
gcagctacct ggagggccag gccgccaagg agttcatcgc ctggctggtg aagggcaggg 660
gccgcaggga cttccctgag gaggtggcca tcgtggagga gctgggccgg cgacacgccg 720
acggcagctt cagcgacgag atgaacacca tcctggacaa cctggccgcg cgcgacttca 780
tcaactggct gatccagacc aagatcaccg atcggaagtg agcgcgctga tatc 834
<210>17
<211>780
<212>DNA
<213>人工序列
<220>
<223>序列描述:依照图11的结构的DNA序列和转译肽序列(参见说明书第35、38页);
<220>
<221>CDS
<222>(11)..(118)
<220>
<221>CDS
<222>(387)..(776)
<400>17
gatatccacc atg gcc ccc gcc gcc tgg ctg agg agc gcc gcc gcc agg 49
Met Ala Pro Ala Ala Trp Leu Arg Ser Ala Ala Ala Arg
1 5 10
gcc ctg ctg cca ccc atg ctg ctg ctg ctg ctg cag ccc cca cct ctg 97
Ala Leu Leu Pro Pro Met Leu Leu Leu Leu Leu Gln Pro Pro Pro Leu
15 20 25
ctg gcc cgg gcc ctg ccc ccg gtgagtgccc gccactcgcc gtccgctcct 148
Leu Ala Arg Ala Leu Pro Pro
30 35
cgctgagggg gcgccgggca cgcgggctgg gcccagcggc gtatccggac gccaagaaac 208
cagagagcca gccagatgcc aaagggccct gccatgtgcc ggtgcccttt ccctctccat 268
ttgccctgcc acacagtggg ctggggttgc acgtgtgttt gctgacaggc cacatctcta 328
actgtgggcc atgtggacct taggcctgac cagaccctca tgtcttcctc cttcccag 386
gac gtg cac cac ctg cac gcc gag agg cgc ggc cct cag ccc tgg cac 434
Asp Val His His Leu His Ala Glu Arg Arg Gly Pro Gln Pro Trp His
40 45 50
gcc gcc ctg cca agc agc cct gcc cct gcc cca gcc acc cag gag gcc 482
Ala Ala Leu Pro Ser Ser Pro Ala Pro Ala Pro Ala Thr Gln Glu Ala
55 60 65
ccc agg cct gcc agc agc ctg agg cca ccc agg tgc ggc gtg cct gat 530
Pro Arg Pro Ala Ser Ser Leu Arg Pro Pro Arg Cys Gly Val Pro Asp
70 75 80
ccc tcc gat ggc ctg agc gct cgg aat cgg cag aag agg cac gcc gag 578
Pro Ser Asp Gly Leu Ser Ala Arg Asn Arg Gln Lys Arg His Ala Glu
85 90 95 100
ggc acc ttc acc tcc gac gtg agc agc tac ctg gag ggc cag gcc gcc 626
Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly Gln Ala Ala
105 110 115
aag gag ttc atc gcc tgg ctg gtg aag ggc agg ggc cgc agg gac ttc 674
Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg Arg Asp Phe
120 125 130
cct gag gag gtg gcc atc gtg gag gag ctg ggc cgg cga cac gcc gac 722
Pro Glu Glu Val Ala Ile Val Glu Glu Leu Gly Arg Arg His Ala Asp
135 140 145
ggc agc ttc agc gac gag atg aac acc atc ctg gac aac ctg gcc gcg 770
Gly Ser Phe Ser Asp Glu Met Asn Thr Ile Leu Asp Asn Leu Ala Ala
150 155 160
cgc tga tatc 780
Arg
165
<210>18
<211>720
<212>DNA
<213>人工序列
<220>
<223>序列描述:依照图1I的DNA序列(参见说明书第39页);
<400>18
gatatccacc atggcccccg ccgcctggct gaggagcgcc gccgccaggg ccctgctgcc 60
acccatgctg ctgctgctgc tgcagccccc acctctgctg gcccgggccc tgcccccggt 120
gagtgcccgc cactcgccgt ccgctcctcg ctgagggggc gccgggcacg cgggctgggc 180
ccagcggcgt atccggacgc caagaaacca gagagccagc cagatgccaa agggccctgc 240
catgtgccgg tgccctttcc ctctccattt gccctgccac acagtgggct ggggttgcac 300
gtgtgtttgc tgacaggcca catctctaac tgtgggccat gtggacctta ggcctgacca 360
gaccctcatg tcttcctcct tcccaggacg tgcaccacct gcacgccgag aggcgcggcc 420
ctcagccctg gcacgccgcc ctgccaagca gccctgcccc tgccccagcc acccaggagg 480
cccccaggcc tgccagcagc ctgaggccac ccaggtgcgg cgtgcctgat ccctccgatg 540
gcctgagcgc tcggaatcgg cagaagaggc acgccgaggg caccttcacc tccgacgtga 600
gcagctacct ggagggccag gccgccaagg agttcatcgc ctggctggtg aagggcaggg 660
acggcagctt cagcgacgag atgaacacca tcctggacaa cctggccgcg cgctgatatc 720
<210>19
<211>716
<212>DNA
<213>人工序列
<220>
<223>序列描述:依照图1m的转译肽的序列和DNA序列(参见说明书第39、40页);
<220>
<221>CDS
<222>(11)..(118)
<220>
<221>CDS
<222>(387)..(710)
<400>19
gatatccacc atg gcc ccc gcc gcc tgg ctg agg agc gcc gcc gcc agg 49
Met Ala Pro Ala Ala Trp Leu Arg Ser Ala Ala Ala Arg
1 5 10
gcc ctg ctg cca ccc atg ctg ctg ctg ctg ctg cag ccc cca cct ctg 97
Ala Leu Leu Pro Pro Met Leu Leu Leu Leu Leu Gln Pro Pro Pro Leu
15 20 25
ctg gcc cgg gcc ctg ccc ccg gtgagtgccc gccactcgcc gtccgctcct 148
Leu Ala Arg Ala Leu Pro Pro
30 35
cgctgagggg gcgccgggca cgcgggctgg gcccagcggc gtatccggac gccaagaaac 208
cagagagcca gccagatgcc aaagggccct gccatgtgcc ggtgcccttt ccctctccat 268
ttgccctgcc acacagtggg ctggggttgc acgtgtgttt gctgacaggc cacatctcta 328
actgtgggcc atgtggacct taggcctgac cagaccctca tgtcttcctc cttcccag 386
gac gtg cac cac ctg cac gcc gag agg cgc ggc cct cag ccc tgg cac 434
Asp Val His His Leu His Ala Glu Arg Arg Gly Pro Gln Pro Trp His
40 45 50
gcc gcc ctg cca agc agc cct gcc cct gcc cca gcc acc cag gag gcc 482
Ala Ala Leu Pro Ser Ser Pro Ala Pro Ala Pro Ala Thr Gln Glu Ala
55 60 65
ccc agg cct gcc agc agc ctg agg cca ccc agg tgc ggc gtg cct gat 530
Pro Arg Pro Ala Ser Ser Leu Arg Pro Pro Arg Cys Gly Val Pro Asp
70 75 80
ccc tcc gat ggc ctg agc gct cgg aat cgg cag aag agg cac gcc gag 578
Pro Ser Asp Gly Leu Ser Ala Arg Asn Arg Gln Lys Arg His Ala Glu
85 90 95 100
ggc acc ttc acc tcc gac gtg agc agc tac ctg gag ggc cag gcc gcc 626
Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly Gln Ala Ala
105 110 115
aag gag ttc atc gcc tgg ctg gtg aag ggc agg ggc cgc agg gac ttc 674
Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg Arg Arg Phe
120 125 130
cct gag gag gtg gcc atc gtg gag gag ctg ggc tga gcgcgc 716
Pro Glu Glu Val Ala Ile Val Glu Glu Leu Gly
135 140
<210>20
<211>716
<212>DNA
<213>人工序列
<220>
<223>序列描述:依照图1m的结构的DNA序列(参见说明书第40页);
<400>20
gatatccacc atggcccccg ccgcctggct gaggagcgcc gccgccaggg ccctgctgcc 60
acccatgctg ctgctgctgc tgcagccccc acctctgctg gcccgggccc tgcccccggt 120
gagtgcccgc cactcgccgt ccgctcctcg ctgagggggc gccgggcacg cgggctgggc 180
ccagcggcgt atccggacgc caagaaacca gagagccagc cagatgccaa agggccctgc 240
catgtgccgg tgccctttcc ctctccattt gccctgccac acagtgggct ggggttgcac 300
gtgtgtttgc tgacaggcca catctctaac tgtgggccat gtggacctta ggcctgacca 360
gaccctcatg tcttcctcct tcccaggacg tgcaccacct gcacgccgag aggcgcggcc 420
ctcagccctg gcacgccgcc ctgccaagca gccctgcccc tgccccagcc acccaggagg 480
cccccaggcc tgccagcagc ctgaggccac ccaggtgcgg cgtgcctgat ccctccgatg 540
gcctgagcgc tcggaatcgg cagaagaggc acgccgaggg caccttcacc tccgacgtga 600
gcagctacct ggagggccag gccgccaagg agttcatcgc ctggctggtg aagggcaggg 660
gccgcaggga cttccctgag gaggtggcca tcgtggagga gctgggctga gcgcgc 716
<210>21
<211>31
<212>PRT
<213>人工序列
<220>
<223>序列描述:人造的(参见说明书第40页);
<400>21
His Gly Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly
20 25 30
<210>22
<211>10
<212>PRT
<213>人工序列
<220>
<223>序列描述:人造的(参见说明书第40页);
<400>22
Arg Arg Asp Phe Pro Glu Glu Val Ala Ile
1 5 10
<210>23
<211>14
<212>PRT
<213>人工序列
<220>
<223>序列描述:人造的(参见说明书第40页);
<400>23
Arg Arg Asp Phe Pro Glu Glu Val Ala Ile Val Glu Glu Leu
1 5 10
<210>24
<211>14
<212>PRT
<213>人工序列
<220>
<223>序列描述:人造的(参见说明书第40页);
<400>24
Arg Arg Asp Phe Pro Glu Glu Val Ala Ile Ala Glu Glu Leu
1 5 10
<210>25
<211>7
<212>PRT
<213>人工序列
<220>
<223>序列描述:从IP2衍生出的部分序列
<400>25
Asp Phe Pro Glu Glu Val Ala
1 5
<210>26
<211>10
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自SEQ ID NO:22的序列,其中头两个丙胺酸被两个精氨酸取代
<400>26
Ala Ala Asp Phe Pro Glu Glu Val Ala Ile
1 5 10
<210>27
<211>14
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自SEQ ID NO:23的序列,其中头两个丙胺酸被两个精氨酸取代
<400>27
Ala Ala Asp Phe Pro Glu Glu Val Ala Ile Val Glu Glu Leu
1 5 10
<210>28
<211>14
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自SEQ ID NO:24的序列,其中头两个丙胺酸被两个精氨酸取代
<400>28
Ala Ala Asp Phe Pro Glu Glu Val Ala Ile Ala Glu Glu Leu
1 5 10
<210>29
<211>15
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自SEQ ID NO:2的序列,其中头两个精氨酸被两个丙胺酸取代
<400>29
Ala Ala Asp Phe Pro Glu Glu Val Ala Ile Val Glu Glu Leu Gly
1 5 10 15
<210>30
<211>15
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自SEQ ID NO:3的序列,其中头两个精氨酸被两个丙胺酸取代
<400>30
Ala Ala Asp Phe Pro Glu Glu Val Ala Ile Ala Glu Glu Leu Gly
1 5 10 15
<210>31
<211>46
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自GLP-1-IP2-片段的序列(鼠)
<400>31
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Ala
20 25 30
Ala Asp Phe Pro Glu Glu Val Ala Ile Ala Glu Glu Leu Gly
35 40 45
<210>32
<211>46
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自GLP-1-IP2-片段的序列(鼠)
<400>32
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
20 25 30
Arg Asp Phe Ala Glu Glu Val Ala Ile Ala Glu Glu Leu Gly
35 40 45
<210>33
<211>46
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自GLP-1-IP2-片段的序列(鼠)
<400>33
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
20 25 30
Arg Asp Ala Ala Ala Ala Val Ala Ile Ala Glu Glu Leu Gly
35 40 45
<210>34
<211>46
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自GLP-1-IP2-片段的序列(鼠)
<400>34
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Ala
20 25 30
Ala Asp Ala Ala Ala Ala Val Ala Ile Ala Ala Ala Leu Gly
35 40 45
<210>35
<211>36
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自GLP-1-IP2-片段的序列(鼠)
<400>35
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
20 25 30
Arg Asp Phe Pro
35
<210>36
<211>40
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自GLP-1-IP2-片段的序列(鼠)
<400>36
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
20 25 30
Arg Asp Phe Pro Glu Glu Val Ala
35 40
<210>37
<211>51
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自GLP-1-IP2-片段的序列(鼠)
<400>37
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
20 25 30
Arg Asp Phe Pro Glu Glu Val Ala Ile Ala Glu Glu Leu Gly Arg Arg
35 40 45
His Ala Cys
50
<210>38
<211>46
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自GLP-1-IP2-片段的序列(人)
<400>38
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Ala
20 25 30
Ala Asp Phe Pro Glu Glu Val Ala Ile Val Glu Glu Leu Gly
35 40 45
<210>39
<211>46
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自GLP-1-IP2-片段的序列(人)
<400>39
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
20 25 30
Arg Asp Phe Ala Glu Glu Val Ala Ile Val Glu Glu Leu Gly
35 40 45
<210>40
<211>46
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自GLP-1-IP2-片段的序列(人)
<400>40
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
20 25 30
Arg Asp Ala Ala Ala Ala Val Ala Ile Val Glu Glu Leu Gly
35 40 45
<210>41
<211>46
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自GLP-1-IP2-片段的序列(人)
<400>41
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Ala
20 25 30
Ala Asp Ala Ala Ala Ala Val Ala Ile Val Ala Ala Leu Gly
35 40 45
<210>42
<211>51
<212>PRT
<213>人工序列
<220>
<223>序列描述:衍生自GLP-1-IP2-片段的序列(人)
<400>42
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly Arg
20 25 30
Arg Asp Phe Pro Glu Glu Val Ala Ile Val Glu Glu Leu Gly Arg Arg
35 40 45
His Ala Cys
50
<210>43
<211>31
<212>PRT
<213>人工序列
<220>
<223>序列描述:依照分子式(I)的序列(参见说明书第1页)
<220>
<221>misc_feature
<222>(31)..(31)
<223>当序列是GLP-1(7-36)醯胺,Xaa=NH2;或当序列是GLP-1(7-37)醯胺时,Xaa=
Gly-OH,
<400>43
His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Xaa
20 25 30
<210>44
<211>31
<212>PRT
<213>人工序列
<220>
<223>序列描述:依照分子式(II)的序列(参见说明书第13页)
<220>
<221>misc_feature
<222>(1)..(1)
<223>Xaa是L-组氨酸,D-组氨酸,去氨基组氨酸,2-氨基-组氨酸,3-羟基-组氨酸,
同源组胺酸,N-乙酰基-组氨酸、a-氟甲基-组氨酸、a-甲基-组氨酸、3-吡啶丙氨酸、
2-吡啶丙氨酸或4-吡啶丙氨酸;
<220>
<221>misc_feature
<222>(2)..(2)
<223>Xaa is Ala,Gly,Val,Leu,Ile,Lys,or Aib;
<220>
<221>misc_feature
<222>(2)..(2)
<223>Xaa可选地是(1-氨基环丙基)羧酸、(1-氨基环丁基)羧酸、(1-氨基环戊基)羧酸、
(1-氨基环己基)羧酸、(1-氨基环庚基)羧酸或(1-氨基环辛基)羧酸
<220>
<221>misc_feature
<222>(10)..(10)
<223>Xaa是Val或Leu;Xaa是Ser、Lys或Arg;
<220>
<221>misc_feature
<222>(12)..(12)
<223>Xaa是Ser、Lys或Arg;
<220>
<221>misc_feature
<222>(13)..(13)
<223>Xaa是Tyr或Gln;
<220>
<221>misc_feature
<222>(14)..(14)
<223>Xaa是Leu或Met;
<220>
<221>misc_feature
<222>(16)..(16)
<223>Xaa是Gly、Glu或Aib;
<220>
<221>misc_feature
<222>(17)..(17)
<223>Xaa是Gln、Glu、Lys或Arg;
<220>
<221>misc_feature
<222>(19)..(19)
<223>Xaa是Ala或Val;
<220>
<221>misc_feature
<222>(20)..(20)
<223>Xaa是Lys、Glu或Arg;
<220>
<221>misc_feature
<222>(21)..(21)
<223>Xaa是Glu或Leu;
<220>
<221>misc_feature
<222>(24)..(24)
<223>Xaa是Ala、Glu或Arg;
<220>
<221>misc_feature
<222>(27)..(27)
<223>Xaa是Val或Lys;
<220>
<221>misc_feature
<222>(28)..(28)
<223>Xaa是Lys、Glu、Asn或Arg;
<220>
<221>misc_feature
<222>(29)..(29)
<223>Xaa是Gly或Aib;
<220>
<221>misc_feature
<222>(30)..(30)
<223>Xaa是Arg、Gly或Lys或醯胺或缺失;
<220>
<221>misc_feature
<222>(31)..(31)
<223>Xaa是Gly、Ala、Glu、Pro、Lys、醯胺或缺失;
<400>44
Xaa Xaa Glu Gly Thr Phe Thr Ser Asp Xaa Ser Xaa Xaa Xaa Glu Xaa
1 5 10 15
Xaa Ala Xaa Xaa Xaa Phe Ile Xaa Trp Leu Xaa Xaa Xaa Xaa Xaa
20 25 30
<210>45
<211>31
<212>PRT
<213>人工序列
<220>
<223>序列描述:依据分子式(III)的序列(参见说明书第13页)
<220>
<221>misc_feature
<222>(1)..(1)
<223>Xaa是是L-组氨酸、D-组氨酸、去氨基-组氨酸、2-氨基-组氨酸、-羟基-组氨酸、
同源组氨酸、N-乙酰基-组氨酸、a-氟甲基-组氨酸、a-甲基-组氨酸、3-吡啶丙氧酸、
2-吡啶丙氨酸或4-吡啶丙氨酸;
<220>
<221>misc_feature
<222>(2)..(2)
<223>Xaa is Ala,Gly,Val,Leu,Ile,Lys or Aib;
<220>
<221>misc_feature
<222>(2)..(2)
<223>Xaa可选地是(1-氨基环丙基)羧酸、(1-氨基环丁基)羧酸、(1-氨基环戊基)羧酸
、(1-氨基环己基)羧酸、(1-氨基环庚基)羧酸或(1-氨基环辛基)羧酸;
<220>
<221>misc_feature
<222>(12)..(12)
<223>Xaa是Ala、Gly、Val、Leu、Ile、Lys或Aib;
<220>
<221>misc_feature
<222>(12)..(12)
<223>Xaa可选地是(1-氨基环丙基)羧酸、(1-氨基环丁基)羧酸、(1-氨基环戊基)羧酸
、(1-氨基环己基)羧酸、(1-氨基环庚基)羧酸或(1-氨基环辛基)羧酸;
<220>
<221>misc_feature
<222>(16)..(16)
<223>Xaa是Gly、Glu或Aib;
<220>
<221>misc_feature
<222>(17)..(17)
<223>Xaa是Gln、Glu、Lys或Arg;
<220>
<221>misc-feature
<222>(20)..(20)
<223>Xaa是Lys、Glu或Arg;
<220>
<221>misc_feature
<222>(24)..(24)
<223>Xaa是Ala、Glu或Arg;
<220>
<221>misc_feature
<222>(28)..(28)
<223>Xaa是Lys、Glu或Arg;
<220>
<221>misc_feature
<222>(29)..(29)
<223>Xaa是Gly或Aib;
<220>
<221>misc_feature
<222>(30)..(30)
<223>Xaa是Arg或Lys、醯胺或缺失;
<220>
<221>misc_feature
<222>(31)..(31)
<223>Xaa是Gly、Ala、Glu或Lys、醯胺或缺失;
<400>45
Xaa Xaa Glu Gly Thr Phe Thr Ser Asp Val Ser Xaa Tyr Leu Glu Xaa
1 5 10 15
Xaa Ala Ala Xaa Glu Phe Ile Xaa Trp Leu Val Xaa Xaa Xaa Xaa
20 25 30
Claims (37)
1. 一种融合肽,其特征在于,包括作为组分(I)的N-端序列,该N-端序列依据分子式II:
Xaa7-Xaa8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Xaa16-Ser-Xaa18-Xaa19-Xaa20-Glu-Xaa22-Xaa23-Ala-Xaa25-Xaa26-Xaa27-Phe-Ile-Xaa3o-Trp-Leu-Xaa33-Xaa34-Xaa35-Xaa36-Xaa37,
其中,Xaa7是L-组氨酸、D-组氨酸、去氨基-组氨酸、2-氨基-组氨酸、3-羟基-组氨酸、同源组氨酸、N-乙酰基-组氨酸、a-氟甲基-组氨酸、a-甲基-组氨酸、3-吡啶丙氨酸、2-吡啶丙氨酸或4-吡啶丙氨酸;Xaa8是Ala、Gly、Val、Leu、Ile、Lys、Aib、(1-氨基环丙基)羧酸、(1-氨基环丁基)羧酸、(1-氨基环戊基)羧酸、(1-氨基环己基)羧酸、(1-氨基环庚基)羧酸或(1-氨基环辛基)羧酸,特别优选Gly;Xaa16是Val或Leu;Xaa是Ser、Lys或Arg;Xaa19是Tyr或Gln;Xaa20是Leu或Met;Xaa22是Gly、Glu或Aib;Xaa23是Gln、Glu、Lys或Arg;Xaa25是Ala或Val;Xaa26是Lys、Glu或Arg;Xaa27是Glu或Leu;Xaa30是Ala、Glu或Arg;Xaa33是Val或Lys;Xaa34是Lys、Glu、Asn或Arg;Xaa35是Gly或Aib;Xaa36是Arg、Gly或Lys或氨基或缺失;Xaa37是Gly、Ala、Glu、Pro、Lys、氨基或缺失;
或分子式III:
Xaa7-Xaa8-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Xaa8-Tyr-Leu-Glu-Xaa22-Xaa23-Ala-Ala-Xaa26-Glu-Phe-lle-Xaa30-Trp-Leu-Val-Xaa34-Xaa35-Xaa36-Xaa37,
其中,Xaa7是L-组氨酸、D-组氨酸、去氨基-组氨酸、2-氨基-组氨酸、-羟基-组氨酸、同源组氨酸、N-乙酰基-组氨酸、a-氟甲基-组氨酸、a-甲基-组氨酸、3-吡啶丙氨酸、2-吡啶丙氨酸或4-吡啶丙氨酸;Xaa8是Ala、Gly、Val、Leu、Ile、Lys、Aib、(1-氨基环丙基)羧酸、(1-氨基环丁基)羧酸、(1-氨基环戊基)羧酸、(1-氨基环己基)羧酸、(1-氨基环庚基)羧酸或(1-氨基环辛基)羧酸;Xaa18是Ser、Lys或Arg;Xaa22是Gly、Glu或Aib;Xaa23是Gln、Glu、Lys或Arg;Xaa26是Lys、Glu或Arg;Xaa34是LYs、Glu或Arg;Xaa35是Gly或Aib;Xaa36是Arg或Lys、氨基或缺失;Xaa37是Gly、Ala、Glu、或Lys、氨基或缺失;
以及作为组分(III)的具有至少9个氨基酸的C-端肽序列或其功能性片段、变异体或衍生物。
2. 根据权利要求1所述的融合肽,其特征在于,包括作为组分(I)的N-端GLP-1(7-35,7-36或7-37)序列以及作为组分(II)的具有至少9个氨基酸的C-端肽序列或其功能性片段、变异体或衍生物。
3. 根据权利要求1所述的融合肽,其特征在于,包括作为组分(I)的N-端序列,该N-端序列依据分子式I:
His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Lys-Gly-Arg-X(I),以及作为组分(III)的具有至少9个氨基酸的C-端肽序列或其功能性片段、变异体或衍生物。
4. 根据权利要求2或3所述的融合肽,其特征在于,所述组分(I)包含与SEQ ID No.:1具有至少80%的序列同源性的序列。
5. 根据权利要求1-4所述的融合肽,其特征在于,所述组分(II)是构成β-回转状结构的肽序列。
6. 根据权利要求1-5中任一项所述的融合肽,其特征在于,所述组分(II)是包含至少一个丙氨酸或脯氨酸残基的肽序列。
7. 根据权利要求1-6中任一项所述的融合肽,其特征在于,所述组分(II)是包含具有β-回转状结构特征的四聚体的肽序列,例如在所述四聚体的位点2具有脯氨酸残基。
8. 根据权利要求1-7中任一项所述的融合肽,其特征在于,所述组分(II)是包含从VAIA、IAEE、PEEV、AEEV、EELG、AAAA、AAVA、AALG、DFPE、AADX、AXDX和XADX中选出的氨基酸序列的肽序列,其中X表示任何氨基酸。
9. 根据权利要求1-8中任一项所述的融合肽,其特征在于,所述组分(II)是通过其N-端氨基酸序列与组分(I)的C-端相连的肽序列,其中所述氨基酸序列选自AA、XA、AX、RR、RX和XR。
10. 根据权利要求1-9中任一项所述的融合肽,其特征在于,所述组分(II)是包含SEQ ID No.:25(DFPEEVA)氨基酸序列的肽序列,或是包含与所述SEQID No.:25(DFPEEVA)具有至少80%同源性的序列的肽序列。
11. 根据权利要求1-10中任一项所述的融合肽,其特征在于,所述组分(II)是包含选自SEQ ID No.:22(RRDFPEEVAI)和SEQ ID No.:26(AADFPEEVAI)的序列的肽序列,或是包含与SEQ ID No.:22(RRDFPEEVAI)和SEQ ID No.:26(AADFPEEVAI)具有至少80%序列同源性的序列的肽序列。
12. 根据权利要求1-11中任一项所述的融合肽,其特征在于,所述组分(II)是包含选自SEQ ID No.:23(RRDFPEEVAIVEEL)、SEQ ID No.24(RRDFPEEVAIAEEL)、SEQ ID No.:27(AADFPEEVAIVEEL)和SEQ ID No.:28(AADFPEEVAIAEEL)的序列的肽序列,或是包含与SEQ ID Nos.:23、24、27或28中任一者至少具有80%序列同源性的序列的肽序列。
13. 根据权利要求1-12中任一项所述的融合肽,其特征在于,所述组分(II)是包含选自SEQ ID No.:2(RRDFPEEVAIVEELG)、SEQ ID No.3(RRDFPEEVAIAEELG)、SEQ ID No.:29(AADFPEEVAIVEELG)和SEQ ID NO.:30(AADFPEEVAIAEELG)的序列的肽序列,或是包含与SEQ ID Nos.:2、3、29或30中任一者至少具有80%序列同源性的序列的肽序列。
14. 根据权利要求1-13中任一项所述的融合肽,其特征在于,所述组分(II)是具有9到30个氨基酸的肽序列,其较佳的是具有9到20个氨基酸的肽序列,更佳地是具有9到15个氨基酸的肽序列。
15. 根据权利要求1-14中任一项所述的融合肽,其特征在于,所述组分(I)和组分(II)直接连接或通过接头序列连接。
16. 根据权利要求15所述的融合肽,其特征在于,所述接头序列具有1到10个氨基酸的长度。
17. 根据权利要求1-16中任一项所述的融合肽,其特征在于,所述融合肽包含有选自
SEQ ID No.:8
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFPEEVAIAEELG)、
SEQ ID No.:12
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFPEEVAIVEELG)、
SEQ ID No.:31
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGAADFPEEVAIAEELG)、
SEQ ID No.:32
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFAEEVAIAEELG)、
SEQ ID No.:33
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDAAAAVAIAEELG)、
SEQ ID No.:34
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGAADAAAAVAIAAALG)、
SEQ ID No.:35
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFP)、
SEQ ID No.:36
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFPEEVA)、
SEQ ID No.:37
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFPEEVAIAEELGRRHAC)、
SEQ ID No.:38
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGAADFPEEVAIVEELG)、
SEQ ID No.:39
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFAEEVAIVEELG)、
SEQ ID No.:40
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDAAAAVAIVEELG)、
SEQ ID No.:41
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGAADAAAAVAIVAALG)和
SEQ ID No.:42
(HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRGRRDFPEEVAIVEELGRRHAC)的序列,或包含有与SEQ ID Nos.:8、12具有至少80%同源性的序列。
18. 根据权利要求1-17中任一项所述的融合肽,其特征在于,所述融合肽包含有与组分(II)的C-端和/或组分(I)的N-端连接另一组分(III)。
19. 根据权利要求18所述的融合肽,其特征在于,所述组分(III)包括至少4个氨基酸残基,较佳地包括10个附加氨基酸残基,更佳地包括至少20个或至少30个附加氨基酸残基。
20. 根据权利要求17或19所述的融合肽,其特征在于,所述组分(III)包括作为胰血糖素中的GLP-2或GLP-1(7-37)的N-端序列的至少4个附加氨基酸残基,优选至少10个附加氨基酸残基,更佳为至少20个附加氨基酸残基。
21. 根据权利要求18-20中任一项所述的融合肽,其特征在于,所述组分(III)包含有SEQ ID Nos.:4或5的序列,或包含有与SEQ ID Nos.:4或5至少具有80%序列同源性的序列。
22. 根据权利要求18-21中任一项所述的融合肽,其特征在于,所述融合肽包含选自SEQ ID No.6、SEQ ID No.7、SEQ ID No.10和SEQ ID No.11的的肽序列,或包含与SEQ ID Nos.:6、7、10或11具有至少80%序列同源性的序列。
23. 根据权利要求1到22中任一项所述的融合肽,其特征在于,所述氨基酸中的至少一个是通过肽主链的修饰,或通过氨基或羧基末端基团的修饰,或通过天然存在的氨基酸的侧链的共价修饰来衍生的。
24. 根据权利要求23所述的融合肽,其特征在于,所述氨基酸中的至少一个是由硫辛酰基或碳氢氧基团衍生的。
25. 根据权利要求23或24所述的融合肽,其特征在于,GLP-1(GLP-1(7))的N-端组氨酸残基在其氨基端和/或在其组胺酰侧链处被化学修饰,特别是被疏水基修饰。
26. 根据权利要求1到25中任一项所述的融合肽,其特征在于,所述融合肽包括作为组分(IV)的载体蛋白质,特别是转铁蛋白或清蛋白。
27. 根据上述权利要求1到26中任一项所述的融合肽,其特征在于,所述组分(I)、(II)和/或(III)的氨基酸序列被反向,且其中所述氨基酸序列至少部分由D氨基酸异构体组成。
28. 一种通过固相肽合成法制备依据前述权利要求1到27中任一项所述的融合肽的方法。
29. 一种用于编码前述权利要求1到22或26中任一项所述的融合肽的核苷酸。
30. 一种包括根据权利要求29所述的核苷酸的媒介。
31. 一种宿主细胞,其特征在于,包括权利要求29所述的外源引入的DNA,特别地包括根据权利要求30所述的媒介,能够表达所述融合肽。
32. 一种用于制备根据前述权利要求1到22或26中任一项所述的融合肽的方法,其中,将经转换包含编码所述融合肽的核苷酸的微生物发酵,并从新获得蛋白质。
33. 一种根据权利要求32制备蛋白质的方法,其中,动物细胞生长在蛋白质从所述细胞中输出的环境下。
34. 根据权利要求1到27中任一项所述的融合肽作为用于治疗人类或动物的药剂。
35. 根据权利要求1到27中任一项所述的融合肽、根据权利要求29所述的核苷酸、根据权利要求30所述的媒介或根据权利要求31所述的宿主细胞用于制备用于治疗I型或II型糖尿病、胰岛素抗药性、体重失调和相关的疾病或生理状况的药剂的用途。
36. 根据权利要求1到27中任一项所述的融合肽、根据权利要求29所述的核苷酸、根据权利要求30所述的媒介或根据权利要求31所述的宿主细胞用于制备用于治疗神经退行性失调或相关疾病或生理状况的药剂的用途。
37. 根据权利要求1到27中任一项所述的融合肽、根据权利要求29所述的核苷酸、根据权利要求30所述的媒介或根据权利要求31所述的宿主细胞用于制备用于治疗与下垂症相关的失调、疾病或生理状况的药剂的用途。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP05020718A EP1767545B1 (en) | 2005-09-22 | 2005-09-22 | GLP-1 (Glucagon-like peptide-1) fusion polypeptides with increased peptidase resistance |
EP05020718.2 | 2005-09-22 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN101273058A true CN101273058A (zh) | 2008-09-24 |
Family
ID=35700191
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CNA2006800350367A Pending CN101273058A (zh) | 2005-09-22 | 2006-09-22 | 增强肽酶抗性的glp-1(胰高血糖素样肽-1)融合多肽 |
Country Status (23)
Country | Link |
---|---|
US (2) | US8431533B2 (zh) |
EP (5) | EP2045265B1 (zh) |
JP (2) | JP5222729B2 (zh) |
KR (1) | KR20080064840A (zh) |
CN (1) | CN101273058A (zh) |
AT (1) | ATE448247T1 (zh) |
AU (1) | AU2006299134B2 (zh) |
BR (1) | BRPI0616107A2 (zh) |
CA (1) | CA2619053A1 (zh) |
CY (1) | CY1109778T1 (zh) |
DE (1) | DE602005017628D1 (zh) |
DK (1) | DK1767545T3 (zh) |
EA (1) | EA013796B1 (zh) |
ES (3) | ES2336575T3 (zh) |
HR (1) | HRP20100062T1 (zh) |
IL (1) | IL188904A0 (zh) |
PL (1) | PL1767545T3 (zh) |
PT (1) | PT1767545E (zh) |
RS (1) | RS51319B (zh) |
SG (1) | SG165414A1 (zh) |
SI (1) | SI1767545T1 (zh) |
WO (1) | WO2007039140A1 (zh) |
ZA (1) | ZA200803488B (zh) |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102363633A (zh) * | 2011-11-16 | 2012-02-29 | 天津拓飞生物科技有限公司 | 胰高血糖素样肽-1突变体多肽及其制备方法、药物组合物和其应用 |
CN102766204A (zh) * | 2011-05-05 | 2012-11-07 | 天津药物研究院 | 胰高血糖素样肽-1突变体多肽及其制备方法和其应用 |
CN104707131A (zh) * | 2013-12-13 | 2015-06-17 | 上海建华精细生物制品有限公司 | 一种药物组合物及其制备方法和用途 |
CN108341880A (zh) * | 2017-01-23 | 2018-07-31 | 天津药物研究院有限公司 | 含有胰高血糖素样肽-1和胰高血糖素的片段类似物的嵌合多肽及其用途 |
CN111278452A (zh) * | 2017-12-26 | 2020-06-12 | 杰优泰德株式会社 | 以阿林肽为活性成分的尿毒症药物 |
CN112661862A (zh) * | 2020-12-25 | 2021-04-16 | 深圳大学 | 一种融合蛋白及其制备方法和应用 |
CN114796462A (zh) * | 2021-12-28 | 2022-07-29 | 北京惠之衡生物科技有限公司 | 一种长效glp-1衍生物的药物制剂 |
WO2023045996A1 (zh) * | 2021-09-26 | 2023-03-30 | 康霖生物科技(杭州)有限公司 | 一种用于糖代谢相关疾病基因治疗的核酸构建体 |
Families Citing this family (30)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
ES2336575T3 (es) | 2005-09-22 | 2010-04-14 | Biocompatibles Uk Limited | Polipeptidos de fusion glp-1 (peptido-1 similar al glucagon) con resistencia aumentada a la peptidasa. |
WO2007049940A1 (en) * | 2005-10-27 | 2007-05-03 | Peptron Co., Ltd | Bioactive substance-blood protein conjugate and stabilization of a bioactive substance using the same |
US20130172274A1 (en) | 2005-12-20 | 2013-07-04 | Duke University | Methods and compositions for delivering active agents with enhanced pharmacological properties |
US8841255B2 (en) | 2005-12-20 | 2014-09-23 | Duke University | Therapeutic agents comprising fusions of vasoactive intestinal peptide and elastic peptides |
DE602006009631D1 (de) | 2006-05-10 | 2009-11-19 | Biocompatibles Uk Ltd | GLP-1 Peptide enthaltende kugelförmige Mikrokapseln, deren Produktion und deren Verwendung |
EP1972349A1 (en) * | 2007-03-21 | 2008-09-24 | Biocompatibles UK Limited | GLP-1 fusion peptides conjugated to polymer(s), their production and use |
EP1975176A1 (en) * | 2007-03-27 | 2008-10-01 | Biocompatibles UK Limited | Novel glp-1 fusion peptides, their production and use |
WO2009158704A2 (en) | 2008-06-27 | 2009-12-30 | Duke University | Therapeutic agents comprising elastin-like peptides |
EP2163243A1 (en) * | 2008-09-12 | 2010-03-17 | Biocompatibles UK Limited | Treatment of acute myocardial infarction (AMI) using encapsulated cells encoding and secreting GLP-1 peptides or analogs thereof |
CN101993485B (zh) | 2009-08-20 | 2013-04-17 | 重庆富进生物医药有限公司 | 促胰岛素分泌肽类似物同源二聚体及其用途 |
MX339559B (es) * | 2010-02-11 | 2016-05-31 | Hoffmann La Roche | Conjugados de igf-i poli(etilenglicol). |
CN103237806A (zh) * | 2010-10-01 | 2013-08-07 | 株式会社Mmt | 具有免疫球蛋白结合能力的肽 |
JP6525456B2 (ja) | 2012-11-20 | 2019-06-05 | メデリス ダイアビーティーズ,エルエルシー | インスリン抵抗性のための改善されたペプチド製剤 |
CN104371019B (zh) | 2013-08-13 | 2019-09-10 | 鸿运华宁(杭州)生物医药有限公司 | 一种能与glp-1r特异性结合的抗体及其与glp-1的融合蛋白质 |
KR20230084337A (ko) | 2014-05-28 | 2023-06-12 | 메더리스 다이어비티즈, 엘엘씨 | 인슐린 저항성에 대한 개선된 펩티드 약제 |
JP6731953B2 (ja) | 2015-02-11 | 2020-07-29 | ジーエムエーエックス バイオファーム エルエルシー. | Glp−1r抗体融合タンパク質の安定な医薬溶液製剤 |
WO2017112889A1 (en) | 2015-12-23 | 2017-06-29 | The Johns Hopkins University | Long-acting glp-1r agonist as a therapy of neurological and neurodegenerative conditions |
CN107818433B (zh) * | 2016-09-14 | 2022-07-05 | 菜鸟智能物流控股有限公司 | 取件方法、物流信息处理方法及装置、系统 |
KR102502040B1 (ko) * | 2016-12-09 | 2023-02-24 | 질랜드 파마 에이/에스 | 아실화 glp-1/glp-2 이중 효능제 |
WO2018104559A1 (en) * | 2016-12-09 | 2018-06-14 | Zealand Pharma A/S | Glp-1/glp-2 dual agonists |
WO2018104558A1 (en) * | 2016-12-09 | 2018-06-14 | Zealand Pharma A/S | Acylated glp-1/glp-2 dual agonists |
IL311775A (en) | 2018-01-03 | 2024-05-01 | Mederis Diabetes Llc | Improved peptide drugs for the treatment of NASH and other disorders |
KR101903564B1 (ko) * | 2018-01-31 | 2018-11-13 | 한국지질자원연구원 | 제지공정에서 발생되는 부산물의 재활용 방법 |
AU2019247936C1 (en) | 2018-04-05 | 2023-06-15 | Sun Pharmaceutical Industries Limited | Novel GLP-1 analogues |
TW202024134A (zh) | 2018-12-21 | 2020-07-01 | 大陸商江蘇恆瑞醫藥股份有限公司 | 雙特異性蛋白 |
KR102349717B1 (ko) * | 2020-03-12 | 2022-01-11 | 주식회사 에스엘메타젠 | 신규 대사증후군 및 그와 관련된 질환 치료용 약학 조성물 |
WO2021182928A1 (ko) * | 2020-03-12 | 2021-09-16 | 주식회사 에스엘메타젠 | 신규 이중 특이성 단백질 및 그의 용도 |
WO2022015039A1 (ko) * | 2020-07-14 | 2022-01-20 | (주)지아이이노베이션 | 글루카곤-유사 펩타이드-1 및 글루카곤-유사 펩타이드-2를 포함하는 융합 단백질 및 이의 용도 |
JP2023553362A (ja) * | 2020-11-27 | 2023-12-21 | ディーアンドディー ファーマテック インコーポレイテッド | ビオチン部分、脂肪酸部分またはその組合せを連結されて有する生物学的活性材料結合体 |
WO2022114908A1 (ko) * | 2020-11-27 | 2022-06-02 | (주)디앤디파마텍 | 비오틴 모이어티, 지방산 모이어티 또는 이의 조합이 결합된 생리활성 물질 접합체의 경구 제형 |
Family Cites Families (83)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US586128A (en) * | 1897-07-13 | Toilet-paper holder | ||
US4352883A (en) * | 1979-03-28 | 1982-10-05 | Damon Corporation | Encapsulation of biological material |
DE2966763D1 (de) | 1979-10-16 | 1984-04-12 | Stocker Winfried | Microanalytic device |
US4596792A (en) | 1981-09-04 | 1986-06-24 | The Regents Of The University Of California | Safe vaccine for hepatitis containing polymerized serum albumin |
JPS5938877A (ja) | 1982-08-30 | 1984-03-02 | Musashi Eng Kk | 紙葉判別方法 |
US4588585A (en) | 1982-10-19 | 1986-05-13 | Cetus Corporation | Human recombinant cysteine depleted interferon-β muteins |
US4737462A (en) | 1982-10-19 | 1988-04-12 | Cetus Corporation | Structural genes, plasmids and transformed cells for producing cysteine depleted muteins of interferon-β |
US4518584A (en) | 1983-04-15 | 1985-05-21 | Cetus Corporation | Human recombinant interleukin-2 muteins |
DD224119A1 (de) | 1983-11-07 | 1985-06-26 | Univ Schiller Jena | Temperierbarer probentraeger |
US4599230A (en) | 1984-03-09 | 1986-07-08 | Scripps Clinic And Research Foundation | Synthetic hepatitis B virus vaccine including both T cell and B cell determinants |
US4599231A (en) | 1984-03-09 | 1986-07-08 | Scripps Clinic And Research Foundation | Synthetic hepatitis B virus vaccine including both T cell and B cell determinants |
US4959314A (en) | 1984-11-09 | 1990-09-25 | Cetus Corporation | Cysteine-depleted muteins of biologically active proteins |
US4608251A (en) | 1984-11-09 | 1986-08-26 | Pitman-Moore, Inc. | LHRH analogues useful in stimulating anti-LHRH antibodies and vaccines containing such analogues |
US5116943A (en) | 1985-01-18 | 1992-05-26 | Cetus Corporation | Oxidation-resistant muteins of Il-2 and other protein |
US4601903A (en) | 1985-05-01 | 1986-07-22 | The United States Of America As Represented By The Department Of Health And Human Services | Vaccine against Neisseria meningitidis Group B serotype 2 invasive disease |
US5144139A (en) * | 1985-08-05 | 1992-09-01 | Biotrack, Inc. | Capillary flow device |
US4687529A (en) * | 1985-08-30 | 1987-08-18 | Miles Laboratories, Inc. | Method of making a reagent test device containing hydrophobic barriers |
CA1292176C (en) | 1985-09-18 | 1991-11-19 | Joel M. Blatt | Volume metering capillary gap device for applying a liquid sample onto a reactive surface |
US5017691A (en) | 1986-07-03 | 1991-05-21 | Schering Corporation | Mammalian interleukin-4 |
US6849708B1 (en) * | 1986-05-05 | 2005-02-01 | The General Hospital Corporation | Insulinotropic hormone and uses thereof |
US4965195A (en) | 1987-10-26 | 1990-10-23 | Immunex Corp. | Interleukin-7 |
US4904584A (en) | 1987-12-23 | 1990-02-27 | Genetics Institute, Inc. | Site-specific homogeneous modification of polypeptides |
WO1991011457A1 (en) | 1990-01-24 | 1991-08-08 | Buckley Douglas I | Glp-1 analogs useful for diabetes treatment |
ES2087323T3 (es) | 1991-02-19 | 1996-07-16 | Takeda Chemical Industries Ltd | Metodo para producir peptidos exentos de cisteina. |
CA2062338A1 (en) | 1991-03-15 | 1992-09-16 | Zia Yassinzadeh | Electronic control cartridge and method of simulating light transmission patterns |
GB9311147D0 (en) | 1993-05-28 | 1993-07-14 | Long Ashton Research Station | Regulation of plant growth |
US5705483A (en) | 1993-12-09 | 1998-01-06 | Eli Lilly And Company | Glucagon-like insulinotropic peptides, compositions and methods |
US5512549A (en) | 1994-10-18 | 1996-04-30 | Eli Lilly And Company | Glucagon-like insulinotropic peptide analogs, compositions, and methods of use |
BE1008987A3 (fr) | 1995-01-06 | 1996-10-01 | Robyn Pierre | Blocs refractaires a canalisations pour sols de fours verriers et installation dans les sols de circuits de refroidissement ou de chauffage, permettant d'ameliorer le controle thermique des sols et la qualite du verre. |
JP3643863B2 (ja) | 1995-08-09 | 2005-04-27 | アークレイ株式会社 | 液体保持具とその製造方法 |
DE69738695D1 (de) | 1996-03-01 | 2008-06-26 | Novo Nordisk As | Peptid zur Appetitzügelung, dessen Zusammensetzungen und Verwendung |
US6165739A (en) * | 1996-03-13 | 2000-12-26 | Compucyte Corporation | Multiple simultaneous testing media |
ATE417622T1 (de) * | 1996-08-08 | 2009-01-15 | Amylin Pharmaceuticals Inc | Regulation gastrointestinaler beweglichkeit |
HU227021B1 (en) * | 1996-08-30 | 2010-05-28 | Novo Nordisk As | Glp-1 derivatives |
AU4863797A (en) | 1996-11-12 | 1998-06-03 | Novo Nordisk A/S | Use of glp-1 peptides |
AU2455199A (en) | 1998-01-12 | 1999-07-26 | Betagene, Inc. | Compositions and methods for regulated secretion from neuroendocrine cell lines |
CA2323160A1 (en) * | 1998-03-06 | 1999-09-10 | Jonathan A. Eppstein | Photothermal structure for biomedical applications, and method therefor |
IL138214A0 (en) * | 1998-03-09 | 2001-10-31 | Zealand Pharmaceuticals As | Pharmacolgically active peptide conjugates having a reduced tendency towards enzymatic hydrolysis |
AU3047299A (en) | 1998-04-13 | 1999-11-01 | Modex Therapeutiques, S.A. | Methods of delivering glp-1 |
US7259136B2 (en) * | 1999-04-30 | 2007-08-21 | Amylin Pharmaceuticals, Inc. | Compositions and methods for treating peripheral vascular disease |
US6267954B1 (en) * | 1999-11-24 | 2001-07-31 | Universite De Paris V Rene-Descartes | Intraocular transplantation of encapsulated cells |
US6706159B2 (en) * | 2000-03-02 | 2004-03-16 | Diabetes Diagnostics | Combined lancet and electrochemical analyte-testing apparatus |
EP1263458B1 (en) | 2000-03-08 | 2005-11-16 | Novo Nordisk A/S | Lowering serum cholesterol |
DE10039643A1 (de) * | 2000-08-14 | 2002-02-28 | Max Planck Gesellschaft | Funktionalisierte Perylentetracarbonsäurediimide |
JP2004528014A (ja) * | 2000-12-07 | 2004-09-16 | イーライ・リリー・アンド・カンパニー | Glp−1融合タンパク質 |
US6821485B2 (en) | 2001-02-09 | 2004-11-23 | Wisconsin Alumni Research Foundation | Method and structure for microfluidic flow guiding |
EP1401480B1 (en) | 2001-02-22 | 2012-11-28 | Novartis AG | Viral vectors encoding endostatin in the treatment of ocular neovascularization |
US20030034147A1 (en) * | 2001-05-31 | 2003-02-20 | Houck Glenn M. | Apparatus which eliminates the need for idling by trucks |
US20030119734A1 (en) | 2001-06-28 | 2003-06-26 | Flink James M. | Stable formulation of modified GLP-1 |
CN1363654A (zh) * | 2001-07-19 | 2002-08-14 | 上海华谊生物技术有限公司 | 生产促胰岛素分泌肽glp-1(7-36)的基因工程菌以及生产glp-1(7-36)的方法 |
EP1430115A1 (en) | 2001-07-27 | 2004-06-23 | Arhus Amt | Immortilized stem cells |
EP1411968B1 (en) | 2001-07-31 | 2008-09-17 | THE GOVERNMENT OF THE UNITED STATES OF AMERICA, as represented by THE SECRETARY, DEPARTMENT OF HEALTH AND HUMAN SERVICES | Glp-1 exendin-4 peptide analogs and uses thereof |
US20040143104A1 (en) * | 2001-08-08 | 2004-07-22 | Wadsworth Samuel C. | Methods of treating diabetes and other blood sugar disorders |
CA2452044A1 (en) | 2001-08-28 | 2003-03-13 | Eli Lilly And Company | Pre-mixes of glp-1 and basal insulin |
US7176278B2 (en) * | 2001-08-30 | 2007-02-13 | Biorexis Technology, Inc. | Modified transferrin fusion proteins |
US7166208B2 (en) * | 2004-03-03 | 2007-01-23 | Stephen Eliot Zweig | Apoenzyme reactivation electrochemical detection method and assay |
AU2002351753A1 (en) * | 2001-12-29 | 2003-07-30 | Novo Nordisk A/S | Combined use of a glp-1 compound and a modulator of diabetic late complications |
WO2003058203A2 (en) | 2002-01-08 | 2003-07-17 | Eli Lilly And Company | Extended glucagon-like peptide-1 analogs |
GB2388898B (en) | 2002-04-02 | 2005-10-05 | Inverness Medical Ltd | Integrated sample testing meter |
US20030191056A1 (en) * | 2002-04-04 | 2003-10-09 | Kenneth Walker | Use of transthyretin peptide/protein fusions to increase the serum half-life of pharmacologically active peptides/proteins |
US20030143113A2 (en) * | 2002-05-09 | 2003-07-31 | Lifescan, Inc. | Physiological sample collection devices and methods of using the same |
CA2497794A1 (en) | 2002-09-06 | 2004-03-18 | Bayer Pharmaceuticals Corporation | Modified glp-1 receptor agonists and their pharmacological methods of use |
JP4548335B2 (ja) * | 2003-03-07 | 2010-09-22 | 味の素株式会社 | 腸管細胞のインスリン産生細胞への変換誘導剤、及び糖尿病治療剤 |
US6969166B2 (en) | 2003-05-29 | 2005-11-29 | 3M Innovative Properties Company | Method for modifying the surface of a substrate |
US7057116B2 (en) | 2003-06-02 | 2006-06-06 | Intel Corporation | Selective reference plane bridge(s) on folded package |
AU2004251145C1 (en) * | 2003-06-12 | 2011-04-14 | Eli Lilly And Company | GLP-1 analog fusion proteins |
EP1498143A1 (en) | 2003-07-18 | 2005-01-19 | Aventis Pharma S.A. | Self-emulsifying and self-microemulsifying formulations for the oral administration of taxoids |
EP1654004A2 (en) | 2003-08-08 | 2006-05-10 | Novo Nordisk A/S | Synthesis and application of new structural well defined branched polymers as conjugating agents for peptides |
CN100580087C (zh) | 2003-10-10 | 2010-01-13 | 诺和诺德医疗保健公司 | 肽的缀合 |
RU2006120077A (ru) * | 2003-12-18 | 2008-01-27 | Ново Нордиск А/С (DK) | Аналоги глюкагоноподобного пептида-1 (glp-1), связанные с альбуминоподобными агентами |
WO2005072216A2 (en) | 2004-01-20 | 2005-08-11 | The Curators Of The University Of Missouri | Supported molecular biofluid viscosity sensors for in vitro and in vivo use |
CN103897066A (zh) | 2004-02-11 | 2014-07-02 | 安米林药品有限责任公司 | 具有可选择特性的杂合多肽 |
BRPI0418539A (pt) | 2004-03-05 | 2007-05-22 | Egomedical Swiss Ag | sistema de teste de analito para a determinação da concentração de um analito em um fluido fisiológico |
AU2005231359A1 (en) | 2004-03-31 | 2005-10-20 | Centocor, Inc. | Human GLP-1 mimetibodies, compositions, methods and uses |
US20080300173A1 (en) * | 2004-07-13 | 2008-12-04 | Defrees Shawn | Branched Peg Remodeling and Glycosylation of Glucagon-Like Peptides-1 [Glp-1] |
CA2574046A1 (en) | 2004-08-13 | 2006-02-16 | Egomedical Technologies Ag | Analyte test system for determining the concentration of an analyte in a physiological or aqueous fluid |
US7442682B2 (en) | 2004-10-19 | 2008-10-28 | Nitto Denko Corporation | Transepithelial delivery of peptides with incretin hormone activities |
WO2007018619A2 (en) | 2005-07-29 | 2007-02-15 | Amprotein Corporation | Chimeric therapeutic agents |
ES2336575T3 (es) | 2005-09-22 | 2010-04-14 | Biocompatibles Uk Limited | Polipeptidos de fusion glp-1 (peptido-1 similar al glucagon) con resistencia aumentada a la peptidasa. |
DE602006009631D1 (de) | 2006-05-10 | 2009-11-19 | Biocompatibles Uk Ltd | GLP-1 Peptide enthaltende kugelförmige Mikrokapseln, deren Produktion und deren Verwendung |
EP1972349A1 (en) * | 2007-03-21 | 2008-09-24 | Biocompatibles UK Limited | GLP-1 fusion peptides conjugated to polymer(s), their production and use |
EP1975176A1 (en) * | 2007-03-27 | 2008-10-01 | Biocompatibles UK Limited | Novel glp-1 fusion peptides, their production and use |
EP2163243A1 (en) | 2008-09-12 | 2010-03-17 | Biocompatibles UK Limited | Treatment of acute myocardial infarction (AMI) using encapsulated cells encoding and secreting GLP-1 peptides or analogs thereof |
-
2005
- 2005-09-22 ES ES05020718T patent/ES2336575T3/es active Active
- 2005-09-22 DK DK05020718.2T patent/DK1767545T3/da active
- 2005-09-22 PL PL05020718T patent/PL1767545T3/pl unknown
- 2005-09-22 SI SI200530887T patent/SI1767545T1/sl unknown
- 2005-09-22 EP EP08021837A patent/EP2045265B1/en active Active
- 2005-09-22 ES ES08021837T patent/ES2397289T3/es active Active
- 2005-09-22 RS RSP-2010/0039A patent/RS51319B/sr unknown
- 2005-09-22 AT AT05020718T patent/ATE448247T1/de active
- 2005-09-22 EP EP05020718A patent/EP1767545B1/en active Active
- 2005-09-22 PT PT05020718T patent/PT1767545E/pt unknown
- 2005-09-22 DE DE602005017628T patent/DE602005017628D1/de active Active
-
2006
- 2006-09-22 CN CNA2006800350367A patent/CN101273058A/zh active Pending
- 2006-09-22 SG SG201006921-9A patent/SG165414A1/en unknown
- 2006-09-22 KR KR1020087009543A patent/KR20080064840A/ko not_active Application Discontinuation
- 2006-09-22 US US11/991,562 patent/US8431533B2/en not_active Expired - Fee Related
- 2006-09-22 BR BRPI0616107-3A patent/BRPI0616107A2/pt not_active IP Right Cessation
- 2006-09-22 ES ES06792228T patent/ES2394218T3/es active Active
- 2006-09-22 ZA ZA200803488A patent/ZA200803488B/xx unknown
- 2006-09-22 EP EP06792228A patent/EP1926748B1/en not_active Not-in-force
- 2006-09-22 JP JP2008531616A patent/JP5222729B2/ja not_active Expired - Fee Related
- 2006-09-22 EP EP09014513A patent/EP2174952A3/en not_active Withdrawn
- 2006-09-22 EP EP10009718A patent/EP2261245A1/en not_active Withdrawn
- 2006-09-22 CA CA002619053A patent/CA2619053A1/en not_active Abandoned
- 2006-09-22 AU AU2006299134A patent/AU2006299134B2/en not_active Ceased
- 2006-09-22 EA EA200800699A patent/EA013796B1/ru not_active IP Right Cessation
- 2006-09-22 WO PCT/EP2006/009226 patent/WO2007039140A1/en active Application Filing
-
2008
- 2008-01-21 IL IL188904A patent/IL188904A0/en unknown
-
2010
- 2010-02-02 HR HR20100062T patent/HRP20100062T1/hr unknown
- 2010-02-11 CY CY20101100139T patent/CY1109778T1/el unknown
-
2012
- 2012-05-29 US US13/482,354 patent/US8853159B2/en not_active Expired - Fee Related
-
2013
- 2013-01-25 JP JP2013012558A patent/JP2013099352A/ja active Pending
Cited By (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102766204A (zh) * | 2011-05-05 | 2012-11-07 | 天津药物研究院 | 胰高血糖素样肽-1突变体多肽及其制备方法和其应用 |
CN102766204B (zh) * | 2011-05-05 | 2014-10-15 | 天津药物研究院 | 胰高血糖素样肽-1突变体多肽及其制备方法和其应用 |
CN102363633A (zh) * | 2011-11-16 | 2012-02-29 | 天津拓飞生物科技有限公司 | 胰高血糖素样肽-1突变体多肽及其制备方法、药物组合物和其应用 |
CN102363633B (zh) * | 2011-11-16 | 2013-11-20 | 天津拓飞生物科技有限公司 | 胰高血糖素样肽-1突变体多肽及其制备方法、药物组合物和其应用 |
CN104707131A (zh) * | 2013-12-13 | 2015-06-17 | 上海建华精细生物制品有限公司 | 一种药物组合物及其制备方法和用途 |
CN108341880A (zh) * | 2017-01-23 | 2018-07-31 | 天津药物研究院有限公司 | 含有胰高血糖素样肽-1和胰高血糖素的片段类似物的嵌合多肽及其用途 |
CN108341880B (zh) * | 2017-01-23 | 2023-07-04 | 天津药物研究院有限公司 | 含有胰高血糖素样肽-1和胰高血糖素的片段类似物的嵌合多肽及其用途 |
CN111278452A (zh) * | 2017-12-26 | 2020-06-12 | 杰优泰德株式会社 | 以阿林肽为活性成分的尿毒症药物 |
CN111278452B (zh) * | 2017-12-26 | 2023-08-29 | 杰优泰德株式会社 | 以阿林肽为活性成分的尿毒症药物 |
CN112661862A (zh) * | 2020-12-25 | 2021-04-16 | 深圳大学 | 一种融合蛋白及其制备方法和应用 |
WO2023045996A1 (zh) * | 2021-09-26 | 2023-03-30 | 康霖生物科技(杭州)有限公司 | 一种用于糖代谢相关疾病基因治疗的核酸构建体 |
CN114796462A (zh) * | 2021-12-28 | 2022-07-29 | 北京惠之衡生物科技有限公司 | 一种长效glp-1衍生物的药物制剂 |
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN101273058A (zh) | 增强肽酶抗性的glp-1(胰高血糖素样肽-1)融合多肽 | |
US8809499B2 (en) | Fusion protein of human fibroblast growth factor-21 and exendin-4 | |
EP1975176A1 (en) | Novel glp-1 fusion peptides, their production and use | |
AU2021204749B2 (en) | Acylated glp-1 derivative | |
KR20060109940A (ko) | 알부민-유사제에 연결된 신규 glp-1 유사체 | |
TW200817432A (en) | Amidated insulin glargine | |
CN102918055A (zh) | 新的胰高血糖素类似物 | |
RU2773242C2 (ru) | Ацилированное производное GLP-1 | |
AU2012202972A1 (en) | GLP-1 fusion peptides, their production and use | |
WO2020228610A1 (zh) | 多肽衍生物及其制备方法 | |
JP2012513981A (ja) | Glp−1アナログおよびその使用 | |
CN102949730A (zh) | 特异结合glp-1受体的药物融合体 | |
NZ622174B2 (en) | Glucagon-like peptide-2 compositions and methods of making and using same |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C12 | Rejection of a patent application after its publication | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20080924 |