CN113429471B - Long-acting GLP-1 polypeptide analogue, and preparation method and application thereof - Google Patents
Long-acting GLP-1 polypeptide analogue, and preparation method and application thereof Download PDFInfo
- Publication number
- CN113429471B CN113429471B CN202110815583.3A CN202110815583A CN113429471B CN 113429471 B CN113429471 B CN 113429471B CN 202110815583 A CN202110815583 A CN 202110815583A CN 113429471 B CN113429471 B CN 113429471B
- Authority
- CN
- China
- Prior art keywords
- gly
- ser
- glu
- polypeptide
- long
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 102400000322 Glucagon-like peptide 1 Human genes 0.000 title claims abstract description 53
- 238000002360 preparation method Methods 0.000 title claims abstract description 7
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 73
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 43
- 229920001184 polypeptide Polymers 0.000 claims abstract description 41
- 206010012601 diabetes mellitus Diseases 0.000 claims abstract description 29
- 208000008589 Obesity Diseases 0.000 claims abstract description 16
- 235000020824 obesity Nutrition 0.000 claims abstract description 15
- 150000001413 amino acids Chemical group 0.000 claims abstract description 10
- 238000004519 manufacturing process Methods 0.000 claims abstract description 3
- 150000001875 compounds Chemical class 0.000 claims description 62
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical group CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 57
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 claims description 55
- 239000003814 drug Substances 0.000 claims description 29
- 239000011347 resin Substances 0.000 claims description 18
- 229920005989 resin Polymers 0.000 claims description 18
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 claims description 14
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 claims description 12
- BDNKZNFMNDZQMI-UHFFFAOYSA-N 1,3-diisopropylcarbodiimide Chemical compound CC(C)N=C=NC(C)C BDNKZNFMNDZQMI-UHFFFAOYSA-N 0.000 claims description 9
- BZLVMXJERCGZMT-UHFFFAOYSA-N Methyl tert-butyl ether Chemical compound COC(C)(C)C BZLVMXJERCGZMT-UHFFFAOYSA-N 0.000 claims description 8
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 claims description 7
- 238000010168 coupling process Methods 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 7
- 230000008878 coupling Effects 0.000 claims description 6
- 238000005859 coupling reaction Methods 0.000 claims description 6
- 230000009089 cytolysis Effects 0.000 claims description 6
- RLSSMJSEOOYNOY-UHFFFAOYSA-N m-cresol Chemical compound CC1=CC=CC(O)=C1 RLSSMJSEOOYNOY-UHFFFAOYSA-N 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 239000003937 drug carrier Substances 0.000 claims description 5
- 239000002904 solvent Substances 0.000 claims description 5
- 238000000605 extraction Methods 0.000 claims description 4
- HCZMHWVFVZAHCR-UHFFFAOYSA-N 2-[2-(2-sulfanylethoxy)ethoxy]ethanethiol Chemical compound SCCOCCOCCS HCZMHWVFVZAHCR-UHFFFAOYSA-N 0.000 claims description 3
- ZHGNHOOVYPHPNJ-UHFFFAOYSA-N Amigdalin Chemical compound FC(F)(F)C(=O)OCC1OC(OCC2OC(OC(C#N)C3=CC=CC=C3)C(OC(=O)C(F)(F)F)C(OC(=O)C(F)(F)F)C2OC(=O)C(F)(F)F)C(OC(=O)C(F)(F)F)C(OC(=O)C(F)(F)F)C1OC(=O)C(F)(F)F ZHGNHOOVYPHPNJ-UHFFFAOYSA-N 0.000 claims description 3
- 239000002671 adjuvant Substances 0.000 claims description 3
- 239000007822 coupling agent Substances 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 238000003776 cleavage reaction Methods 0.000 claims description 2
- 239000012043 crude product Substances 0.000 claims description 2
- 238000001556 precipitation Methods 0.000 claims description 2
- 230000002194 synthesizing effect Effects 0.000 claims description 2
- 230000015572 biosynthetic process Effects 0.000 abstract description 17
- 238000003786 synthesis reaction Methods 0.000 abstract description 17
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 abstract description 10
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 abstract description 4
- 230000000857 drug effect Effects 0.000 abstract description 3
- 239000004471 Glycine Substances 0.000 abstract description 2
- 125000003277 amino group Chemical group 0.000 abstract description 2
- 230000003321 amplification Effects 0.000 abstract description 2
- 230000008901 benefit Effects 0.000 abstract description 2
- 230000002503 metabolic effect Effects 0.000 abstract description 2
- 238000003199 nucleic acid amplification method Methods 0.000 abstract description 2
- 125000005313 fatty acid group Chemical group 0.000 abstract 1
- 101800000224 Glucagon-like peptide 1 Proteins 0.000 description 38
- 210000004369 blood Anatomy 0.000 description 38
- 239000008280 blood Substances 0.000 description 38
- 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 33
- 241000699670 Mus sp. Species 0.000 description 33
- 239000008103 glucose Substances 0.000 description 33
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 21
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 18
- 230000000694 effects Effects 0.000 description 18
- 230000037396 body weight Effects 0.000 description 17
- 229940079593 drug Drugs 0.000 description 16
- 102000007446 Glucagon-Like Peptide-1 Receptor Human genes 0.000 description 15
- 108010086246 Glucagon-Like Peptide-1 Receptor Proteins 0.000 description 15
- 239000000047 product Substances 0.000 description 14
- 239000000243 solution Substances 0.000 description 14
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 13
- 241000700159 Rattus Species 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 12
- -1 fmoc-Gly-OH Chemical compound 0.000 description 12
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 11
- 206010022489 Insulin Resistance Diseases 0.000 description 10
- 241000699666 Mus <mouse, genus> Species 0.000 description 10
- 125000003275 alpha amino acid group Chemical group 0.000 description 10
- 229940125898 compound 5 Drugs 0.000 description 10
- 238000007410 oral glucose tolerance test Methods 0.000 description 10
- 229940125782 compound 2 Drugs 0.000 description 9
- 238000010254 subcutaneous injection Methods 0.000 description 9
- 239000007929 subcutaneous injection Substances 0.000 description 9
- REITVGIIZHFVGU-IBGZPJMESA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-[(2-methylpropan-2-yl)oxy]propanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](COC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 REITVGIIZHFVGU-IBGZPJMESA-N 0.000 description 8
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 8
- 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 8
- 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 8
- 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 8
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 8
- FBNPMTNBFFAMMH-AVGNSLFASA-N Leu-Val-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N FBNPMTNBFFAMMH-AVGNSLFASA-N 0.000 description 8
- FBNPMTNBFFAMMH-UHFFFAOYSA-N Leu-Val-Arg Natural products CC(C)CC(N)C(=O)NC(C(C)C)C(=O)NC(C(O)=O)CCCN=C(N)N FBNPMTNBFFAMMH-UHFFFAOYSA-N 0.000 description 8
- 235000001014 amino acid Nutrition 0.000 description 8
- 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 description 8
- 239000000463 material Substances 0.000 description 8
- 238000011160 research Methods 0.000 description 8
- 108010080629 tryptophan-leucine Proteins 0.000 description 8
- 229940024606 amino acid Drugs 0.000 description 7
- 238000005516 engineering process Methods 0.000 description 7
- 238000012544 monitoring process Methods 0.000 description 7
- 210000003462 vein Anatomy 0.000 description 7
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 6
- 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 description 6
- 102000017011 Glycated Hemoglobin A Human genes 0.000 description 6
- 230000001270 agonistic effect Effects 0.000 description 6
- 230000037406 food intake Effects 0.000 description 6
- 235000012631 food intake Nutrition 0.000 description 6
- 238000003304 gavage Methods 0.000 description 6
- 108091005995 glycated hemoglobin Proteins 0.000 description 6
- 210000004185 liver Anatomy 0.000 description 6
- 238000012986 modification Methods 0.000 description 6
- 230000004048 modification Effects 0.000 description 6
- 235000010436 thaumatin Nutrition 0.000 description 6
- 239000000892 thaumatin Substances 0.000 description 6
- 238000005406 washing Methods 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- NDKDFTQNXLHCGO-UHFFFAOYSA-N 2-(9h-fluoren-9-ylmethoxycarbonylamino)acetic acid Chemical compound C1=CC=C2C(COC(=O)NCC(=O)O)C3=CC=CC=C3C2=C1 NDKDFTQNXLHCGO-UHFFFAOYSA-N 0.000 description 5
- MTCFGRXMJLQNBG-UWTATZPHSA-N D-Serine Chemical compound OC[C@@H](N)C(O)=O MTCFGRXMJLQNBG-UWTATZPHSA-N 0.000 description 5
- 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 5
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 5
- 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 5
- 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 5
- PLQWGQUNUPMNOD-KKUMJFAQSA-N Ser-Tyr-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(O)=O PLQWGQUNUPMNOD-KKUMJFAQSA-N 0.000 description 5
- 125000000539 amino acid group Chemical group 0.000 description 5
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 5
- 230000003914 insulin secretion Effects 0.000 description 5
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 5
- 239000006166 lysate Substances 0.000 description 5
- 102000004877 Insulin Human genes 0.000 description 4
- 108090001061 Insulin Proteins 0.000 description 4
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 4
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 239000002552 dosage form Substances 0.000 description 4
- 201000010063 epididymitis Diseases 0.000 description 4
- 239000003925 fat Substances 0.000 description 4
- 229940125396 insulin Drugs 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 229910052757 nitrogen Inorganic materials 0.000 description 4
- 239000008194 pharmaceutical composition Substances 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- 238000006467 substitution reaction Methods 0.000 description 4
- DTQVDTLACAAQTR-UHFFFAOYSA-N trifluoroacetic acid Substances OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 4
- 239000003981 vehicle Substances 0.000 description 4
- OTKXCALUHMPIGM-FQEVSTJZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-5-[(2-methylpropan-2-yl)oxy]-5-oxopentanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCC(=O)OC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 OTKXCALUHMPIGM-FQEVSTJZSA-N 0.000 description 3
- DBTMQODRSDEGRZ-UHFFFAOYSA-N 9h-fluoren-9-ylmethyl n-(2-oxoethyl)carbamate Chemical compound C1=CC=C2C(COC(=O)NCC=O)C3=CC=CC=C3C2=C1 DBTMQODRSDEGRZ-UHFFFAOYSA-N 0.000 description 3
- 108010088751 Albumins Proteins 0.000 description 3
- 102000009027 Albumins Human genes 0.000 description 3
- 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 3
- YWAQATDNEKZFFK-BYPYZUCNSA-N Gly-Gly-Ser Chemical compound NCC(=O)NCC(=O)N[C@@H](CO)C(O)=O YWAQATDNEKZFFK-BYPYZUCNSA-N 0.000 description 3
- 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 3
- JMSONHOUHFDOJH-GUBZILKMSA-N His-Ser-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CN=CN1 JMSONHOUHFDOJH-GUBZILKMSA-N 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- DLSWIYLPEUIQAV-UHFFFAOYSA-N Semaglutide Chemical compound CCC(C)C(NC(=O)C(Cc1ccccc1)NC(=O)C(CCC(O)=O)NC(=O)C(CCCCNC(=O)COCCOCCNC(=O)COCCOCCNC(=O)CCC(NC(=O)CCCCCCCCCCCCCCCCC(O)=O)C(O)=O)NC(=O)C(C)NC(=O)C(C)NC(=O)C(CCC(N)=O)NC(=O)CNC(=O)C(CCC(O)=O)NC(=O)C(CC(C)C)NC(=O)C(Cc1ccc(O)cc1)NC(=O)C(CO)NC(=O)C(CO)NC(=O)C(NC(=O)C(CC(O)=O)NC(=O)C(CO)NC(=O)C(NC(=O)C(Cc1ccccc1)NC(=O)C(NC(=O)CNC(=O)C(CCC(O)=O)NC(=O)C(C)(C)NC(=O)C(N)Cc1cnc[nH]1)C(C)O)C(C)O)C(C)C)C(=O)NC(C)C(=O)NC(Cc1c[nH]c2ccccc12)C(=O)NC(CC(C)C)C(=O)NC(C(C)C)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CCCNC(N)=N)C(=O)NCC(O)=O DLSWIYLPEUIQAV-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 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 3
- 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 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000012790 confirmation Methods 0.000 description 3
- 150000004665 fatty acids Chemical group 0.000 description 3
- 239000012091 fetal bovine serum Substances 0.000 description 3
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 230000000405 serological effect Effects 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- OYXZPXVCRAAKCM-SANMLTNESA-N (2s)-2-[(2-methylpropan-2-yl)oxycarbonylamino]-3-(1-tritylimidazol-4-yl)propanoic acid Chemical compound C1=NC(C[C@H](NC(=O)OC(C)(C)C)C(O)=O)=CN1C(C=1C=CC=CC=1)(C=1C=CC=CC=1)C1=CC=CC=C1 OYXZPXVCRAAKCM-SANMLTNESA-N 0.000 description 2
- HNICLNKVURBTKV-NDEPHWFRSA-N (2s)-5-[[amino-[(2,2,4,6,7-pentamethyl-3h-1-benzofuran-5-yl)sulfonylamino]methylidene]amino]-2-(9h-fluoren-9-ylmethoxycarbonylamino)pentanoic acid Chemical compound C12=CC=CC=C2C2=CC=CC=C2C1COC(=O)N[C@H](C(O)=O)CCCN=C(N)NS(=O)(=O)C1=C(C)C(C)=C2OC(C)(C)CC2=C1C HNICLNKVURBTKV-NDEPHWFRSA-N 0.000 description 2
- LZOLWEQBVPVDPR-VLIAUNLRSA-N (2s,3r)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-[(2-methylpropan-2-yl)oxy]butanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H]([C@H](OC(C)(C)C)C)C(O)=O)C3=CC=CC=C3C2=C1 LZOLWEQBVPVDPR-VLIAUNLRSA-N 0.000 description 2
- GOPWHXPXSPIIQZ-FQEVSTJZSA-N (4s)-4-(9h-fluoren-9-ylmethoxycarbonylamino)-5-[(2-methylpropan-2-yl)oxy]-5-oxopentanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCC(O)=O)C(=O)OC(C)(C)C)C3=CC=CC=C3C2=C1 GOPWHXPXSPIIQZ-FQEVSTJZSA-N 0.000 description 2
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 2
- 208000007342 Diabetic Nephropathies Diseases 0.000 description 2
- 108090000331 Firefly luciferases Proteins 0.000 description 2
- 108010004460 Gastric Inhibitory Polypeptide Proteins 0.000 description 2
- QXUPRMQJDWJDFR-NRPADANISA-N Glu-Val-Ser Chemical compound CC(C)[C@H](NC(=O)[C@@H](N)CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O QXUPRMQJDWJDFR-NRPADANISA-N 0.000 description 2
- 108010088406 Glucagon-Like Peptides Proteins 0.000 description 2
- BCCRXDTUTZHDEU-VKHMYHEASA-N Gly-Ser Chemical compound NCC(=O)N[C@@H](CO)C(O)=O BCCRXDTUTZHDEU-VKHMYHEASA-N 0.000 description 2
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 2
- QIJVAFLRMVBHMU-KKUMJFAQSA-N Lys-Asp-Phe Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O QIJVAFLRMVBHMU-KKUMJFAQSA-N 0.000 description 2
- YNAVUWVOSKDBBP-UHFFFAOYSA-N Morpholine Chemical compound C1COCCN1 YNAVUWVOSKDBBP-UHFFFAOYSA-N 0.000 description 2
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 2
- 108010052090 Renilla Luciferases Proteins 0.000 description 2
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 2
- 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 2
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Chemical compound CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 210000000227 basophil cell of anterior lobe of hypophysis Anatomy 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 230000001684 chronic effect Effects 0.000 description 2
- 229940125904 compound 1 Drugs 0.000 description 2
- 229940126214 compound 3 Drugs 0.000 description 2
- 238000006482 condensation reaction Methods 0.000 description 2
- 230000001808 coupling effect Effects 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 208000033679 diabetic kidney disease Diseases 0.000 description 2
- 230000002526 effect on cardiovascular system Effects 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 125000003630 glycyl group Chemical group [H]N([H])C([H])([H])C(*)=O 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
- 201000001421 hyperglycemia Diseases 0.000 description 2
- 230000002218 hypoglycaemic effect Effects 0.000 description 2
- 238000010253 intravenous injection Methods 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 239000000644 isotonic solution Substances 0.000 description 2
- 210000004731 jugular vein Anatomy 0.000 description 2
- 238000004949 mass spectrometry Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 208000030159 metabolic disease Diseases 0.000 description 2
- 125000001360 methionine group Chemical group N[C@@H](CCSC)C(=O)* 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 230000004526 pharmaceutical effect Effects 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 230000036470 plasma concentration Effects 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000005086 pumping Methods 0.000 description 2
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 108010060325 semaglutide Proteins 0.000 description 2
- 229950011186 semaglutide Drugs 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- KQKPFRSPSRPDEB-UHFFFAOYSA-N sumatriptan Chemical compound CNS(=O)(=O)CC1=CC=C2NC=C(CCN(C)C)C2=C1 KQKPFRSPSRPDEB-UHFFFAOYSA-N 0.000 description 2
- 229960003708 sumatriptan Drugs 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- ADOHASQZJSJZBT-SANMLTNESA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-[1-[(2-methylpropan-2-yl)oxycarbonyl]indol-3-yl]propanoic acid Chemical compound C12=CC=CC=C2N(C(=O)OC(C)(C)C)C=C1C[C@@H](C(O)=O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21 ADOHASQZJSJZBT-SANMLTNESA-N 0.000 description 1
- JAUKCFULLJFBFN-VWLOTQADSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-3-[4-[(2-methylpropan-2-yl)oxy]phenyl]propanoic acid Chemical compound C1=CC(OC(C)(C)C)=CC=C1C[C@@H](C(O)=O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21 JAUKCFULLJFBFN-VWLOTQADSA-N 0.000 description 1
- FODJWPHPWBKDON-IBGZPJMESA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-4-[(2-methylpropan-2-yl)oxy]-4-oxobutanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CC(=O)OC(C)(C)C)C(O)=O)C3=CC=CC=C3C2=C1 FODJWPHPWBKDON-IBGZPJMESA-N 0.000 description 1
- WDGICUODAOGOMO-DHUJRADRSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-5-oxo-5-(tritylamino)pentanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC1C2=CC=CC=C2C2=CC=CC=C21)CC(=O)NC(C=1C=CC=CC=1)(C=1C=CC=CC=1)C1=CC=CC=C1 WDGICUODAOGOMO-DHUJRADRSA-N 0.000 description 1
- OJBNDXHENJDCBA-QFIPXVFZSA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)-6-(prop-2-enoxycarbonylamino)hexanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](CCCCNC(=O)OCC=C)C(=O)O)C3=CC=CC=C3C2=C1 OJBNDXHENJDCBA-QFIPXVFZSA-N 0.000 description 1
- 125000001431 2-aminoisobutyric acid group Chemical group [#6]C([#6])(N*)C(*)=O 0.000 description 1
- WDBQJSCPCGTAFG-QHCPKHFHSA-N 4,4-difluoro-N-[(1S)-3-[4-(3-methyl-5-propan-2-yl-1,2,4-triazol-4-yl)piperidin-1-yl]-1-pyridin-3-ylpropyl]cyclohexane-1-carboxamide Chemical compound FC1(CCC(CC1)C(=O)N[C@@H](CCN1CCC(CC1)N1C(=NN=C1C)C(C)C)C=1C=NC=CC=1)F WDBQJSCPCGTAFG-QHCPKHFHSA-N 0.000 description 1
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 208000017667 Chronic Disease Diseases 0.000 description 1
- 208000032928 Dyslipidaemia Diseases 0.000 description 1
- 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 1
- 208000004930 Fatty Liver Diseases 0.000 description 1
- 229940089838 Glucagon-like peptide 1 receptor agonist Drugs 0.000 description 1
- 108010023302 HDL Cholesterol Proteins 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- 108010028554 LDL Cholesterol Proteins 0.000 description 1
- 101150046735 LEPR gene Proteins 0.000 description 1
- 101150063827 LEPROT gene Proteins 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 208000017170 Lipid metabolism disease Diseases 0.000 description 1
- 108010019598 Liraglutide Proteins 0.000 description 1
- YSDQQAXHVYUZIW-QCIJIYAXSA-N Liraglutide Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCNC(=O)CC[C@H](NC(=O)CCCCCCCCCCCCCCC)C(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)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](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O)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=C(O)C=C1 YSDQQAXHVYUZIW-QCIJIYAXSA-N 0.000 description 1
- PGBPWPTUOSCNLE-JYJNAYRXSA-N Lys-Gln-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCCCN)N PGBPWPTUOSCNLE-JYJNAYRXSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 1
- LFZAGIJXANFPFN-UHFFFAOYSA-N N-[3-[4-(3-methyl-5-propan-2-yl-1,2,4-triazol-4-yl)piperidin-1-yl]-1-thiophen-2-ylpropyl]acetamide Chemical compound C(C)(C)C1=NN=C(N1C1CCN(CC1)CCC(C=1SC=CC=1)NC(C)=O)C LFZAGIJXANFPFN-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010067482 No adverse event Diseases 0.000 description 1
- 208000031662 Noncommunicable disease Diseases 0.000 description 1
- 206010053159 Organ failure Diseases 0.000 description 1
- 206010033307 Overweight Diseases 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- XHWCDRUPDNSDAZ-XKBZYTNZSA-N Thr-Ser-Glu Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N)O XHWCDRUPDNSDAZ-XKBZYTNZSA-N 0.000 description 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 239000000556 agonist Substances 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 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 1
- 230000000903 blocking effect Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 230000007211 cardiovascular event Effects 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 150000001793 charged compounds Chemical class 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000009693 chronic damage Effects 0.000 description 1
- 238000011960 computer-aided design Methods 0.000 description 1
- 208000012696 congenital leptin deficiency Diseases 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 239000008298 dragée Substances 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 230000007515 enzymatic degradation Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 239000012065 filter cake Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 208000020694 gallbladder disease Diseases 0.000 description 1
- 210000004211 gastric acid Anatomy 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 230000010030 glucose lowering effect Effects 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 125000000291 glutamic acid group Chemical group N[C@@H](CCC(O)=O)C(=O)* 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 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 1
- 239000000859 incretin Substances 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000011835 investigation 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
- 125000000741 isoleucyl group Chemical group [H]N([H])C(C(C([H])([H])[H])C([H])([H])C([H])([H])[H])C(=O)O* 0.000 description 1
- GCHPUFAZSONQIV-UHFFFAOYSA-N isovaline Chemical compound CCC(C)(N)C(O)=O GCHPUFAZSONQIV-UHFFFAOYSA-N 0.000 description 1
- 208000017169 kidney disease Diseases 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 125000001909 leucine group Chemical group [H]N(*)C(C(*)=O)C([H])([H])C(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 1
- 239000012669 liquid formulation Substances 0.000 description 1
- 229960002701 liraglutide Drugs 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 238000013227 male C57BL/6J mice Methods 0.000 description 1
- COTNUBDHGSIOTA-UHFFFAOYSA-N meoh methanol Chemical compound OC.OC COTNUBDHGSIOTA-UHFFFAOYSA-N 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 239000000693 micelle Substances 0.000 description 1
- 239000004530 micro-emulsion Substances 0.000 description 1
- 229910000403 monosodium phosphate Inorganic materials 0.000 description 1
- 235000019799 monosodium phosphate Nutrition 0.000 description 1
- 208000001022 morbid obesity Diseases 0.000 description 1
- 239000007922 nasal spray Substances 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 230000004768 organ dysfunction Effects 0.000 description 1
- 229910052763 palladium Inorganic materials 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 229940021222 peritoneal dialysis isotonic solution Drugs 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 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
- COLNVLDHVKWLRT-QMMMGPOBSA-N phenylalanine group Chemical group N[C@@H](CC1=CC=CC=C1)C(=O)O COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 210000004896 polypeptide structure Anatomy 0.000 description 1
- 238000002953 preparative HPLC Methods 0.000 description 1
- GCYXWQUSHADNBF-AAEALURTSA-N preproglucagon 78-108 Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1N=CNC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=CC=C1 GCYXWQUSHADNBF-AAEALURTSA-N 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 125000003607 serino group Chemical group [H]N([H])[C@]([H])(C(=O)[*])C(O[H])([H])[H] 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 239000004017 serum-free culture medium Substances 0.000 description 1
- 201000002859 sleep apnea Diseases 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000008174 sterile solution Substances 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 238000005556 structure-activity relationship Methods 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 239000007940 sugar coated tablet Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 125000000341 threoninyl group Chemical group [H]OC([H])(C([H])([H])[H])C([H])(N([H])[H])C(*)=O 0.000 description 1
- 125000000430 tryptophan group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C2=C([H])C([H])=C([H])C([H])=C12 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 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
- 238000001291 vacuum drying Methods 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 239000013585 weight reducing agent Substances 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
- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Diabetes (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Hematology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Endocrinology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Obesity (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Child & Adolescent Psychology (AREA)
- Zoology (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Emergency Medicine (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention belongs to the technical field of biochemistry, and particularly relates to a long-acting GLP-1 polypeptide analogue for treating or preventing diabetes or obesity, and a preparation method and application thereof. Solves the problems of short half-life period and insufficient metabolic stability of complex polypeptide. The amino acid sequence structure of the long-acting GLP-1 polypeptide analogue of the invention is as follows: the 20 th Lys is connected with a short peptide chain of a side arm in a mode of: the amino group of the short peptide chain of the side arm of the Lys at the 20 th position and the carboxyl group of the glycine of the short peptide chain of the side arm form an amido bond; the amino acid of the end Z of the short peptide chain of the side arm is connected with a fatty acid substituent. The long-acting GLP-1 polypeptide analogue has the advantages of long half-life period, high synthesis yield, good stability, easy amplification production and low cost, and has good drug effect on treating diabetes and reducing weight.
Description
Technical Field
The invention belongs to the technical field of biochemistry, and particularly relates to long-acting GLP-1 polypeptide analogues for treating or preventing diabetes or obesity, and a preparation method and application thereof.
Background
Diabetes is a chronic disease in which the body is at high blood sugar level for a long time to cause sugar metabolism disorder of the body, and the disease is mainly characterized in that: chronic hyperglycemia, and at the same time insulin secretion deficiency or insulin dysfunction, further causes chronic damage to various organs by affecting the metabolism of fats, carbohydrates and proteins, resulting in gradual organ dysfunction and even organ failure.
Over the past three decades, the number of diabetics worldwide has increased by three times. Worldwide, nearly 9% of adults suffer from type 2 diabetes (T2 DM). The advent of T2DM and its complications has greatly exacerbated the worldwide risk of disability and death. For example, the "2013 global risk for disease study" identified diabetes (all forms) as the ninth leading cause of decreased life expectancy. Based on the results of many investigations, it has been shown that diabetes is not effectively controlled in developed countries in europe and america, or in developing countries like china. Diabetes has become another important chronic non-infectious disease seriously endangering human health after cardiovascular and cerebrovascular diseases and tumors.
Obesity and diabetes belong to metabolic diseases, and are closely related to the occurrence of diabetes. Diabetes is mainly caused by the decline of islet beta cell function and insulin resistance, and obesity is a key factor of insulin resistance. The obesity patient is easy to generate insulin resistance due to excessive weight and high fat content, and the insulin resistance can not enable insulin in the body to play a corresponding role in reducing blood sugar. Insulin is the only blood sugar-lowering hormone in the body, the body must increase the insulin secretion capacity of islet beta cells for controlling blood sugar, and diabetes can occur when the blood sugar is still not normal due to the increase of insulin secretion, so that obesity is the root cause of insulin resistance and is also an important reason for the occurrence of diabetes.
An interesting phenomenon observed in McIntyre and ellick et al in the 60's of the 20 th century, the promotion of insulin secretion by oral glucose was significantly higher than that by intravenous injection, an effect known as the "incretin effect", and glucagon-like peptide-1 (GLP-1) and glucose-dependent insulin-releasing peptide (GIP) were subsequently found in extracts of the small intestinal mucosa. GLP-1 is a hormone that induces insulin secretion and has beneficial effects on a variety of vital organs including pancreas, heart, liver, etc. GLP-1 receptor (GLP-1R) agonist drugs have the advantages of effectively controlling blood sugar, obviously reducing incidence rate of hypoglycemic events, obviously reducing weight and reducing risk of cardiovascular events. However, due to the characteristics of a specific polypeptide structure of the GLP-1 medicament, the GLP-1 medicament has instability, can be degraded by gastric acid after being orally taken, can be basically administrated only by subcutaneous injection, and has short half life.
With the intensive research on diabetes and the treatment thereof, GLP-1 receptor agonists such as liraglutide, somaglutide and the like have been approved for marketing in recent years. Wherein, the affinity of the increased carbon chain of the somagluteptide to albumin is greatly enhanced, and the elimination of the somagluteptide to albumin is greatly slowed down. The two reconstruction parts prolong the half-life period of the rat to about 8h, and only one subcutaneous injection is needed clinically every week.
Nevertheless, how to solve the problems of short half-life and insufficient metabolic stability of polypeptides, especially complex polypeptides, in a breakthrough manner remains a major scientific and core problem in the field; the development of the ultra-long-acting polypeptide drug molecule modification technology is a key and is also a bottleneck to be broken through internationally in the field of research.
Disclosure of Invention
In view of the above problems, it is an object of the present invention to provide a new class of GLP-1 polypeptide analogues with a longer duration.
It is a further object of the present invention to provide a method for the preparation of such long-acting GLP-1 polypeptide analogues.
It is a further object of the present invention to provide a composition comprising the above-described long-acting GLP-1 polypeptide analog.
Still another object of the present invention is to provide uses of the above GLP-1 polypeptide analogs.
According to a specific embodiment of the invention, the long-acting GLP-1 polypeptide analogue has the following amino acid sequence:
His-Xaa2-Glu-Gly-Thr-Phe-Thr-Ser-Xaa9-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys{(Gly) x -(Ser-Gly) y -Z-CO(CH 2 ) n CO 2 H}-Xaa21-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly
wherein:
xaa2 is Aib or D-Ser;
xaa9 is Asp or Glu;
xaa21 is Glu or Gln or Asp;
z is gamma Glu or Asp;
x is 1, 2, 3 or 4;
y is 1, 2, 3 or 4;
n is an integer from 12 to 20, i.e. n is 12, 13, 14, 15, 16, 17, 18, 19 or 20.
Wherein, the first and the second end of the pipe are connected with each other,
His-Xaa2-Glu-Gly-Thr-Phe-Thr-Ser-Xaa9-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Xaa21-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly as main peptide chain;
(Gly) x -(Ser-Gly) y -Z-CO(CH 2 ) n CO 2 h is a side arm structure;
the side arm structure comprises a side arm short peptide chain- (Gly) x- (Ser-Gly) y-Z-and a fatty acid substituent-CO (CH) connected with the side arm short peptide chain- (Gly) 2 ) n CO 2 H。
The side chain amino of the 20 th Lys in the main peptide chain amino acid sequence is connected with a side arm short peptide chain structure in an amido bond mode formed by the side chain amino and the carboxyl of the other end glycine residue;
further, the amino group of the "Z" amino acid residue at the end of the "side arm" short peptide chain is linked to a fatty acid substituent by forming an amide bond with a carboxyl group.
The carboxyl terminal of the main peptide chain amino acid sequence is not modified or is modified by amino to form-CONH 2 A group.
Preferably, Z is gammaglu.
Preferably, x is 2, y is 1, 2 or 3, and in particular, in the above amino acid sequence, x is 2, y is 1, or x is 2, y is 2, or x is 2, y is 3.
Preferably, in the amino acid sequence structure: n is any integer of 14-18, namely n is 14, 15, 16, 17 or 18.
According to a specific embodiment of the invention, a long-acting GLP-1 polypeptide analog has an amino acid sequence wherein Z is gamma Glu, x is 2, y is 1, 2 or 3, n is 18, and the sequence is as follows: ,
His-Xaa2-Glu-Gly-Thr-Phe-Thr-Ser-Xaa9-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys{(Gly) 2 -(Ser-Gly) y -γGlu-CO(CH 2 ) 18 CO 2 H}-Xaa21-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly。
preferably, the GLP-1 polypeptide analogue is any one of the following compounds:
compound 1 (SEQ ID NO. 1):
His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys(Gly-Gly-Ser-Gly-γ-Glu-CO(CH 2 ) 18 CO 2 H)-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly;
the abbreviation is: H-Aib-EGTFTSDVSSYLEGQAAK (GGSG-gamma-E-CO (CH) 2 ) 18 CO 2 H)EFIAWLVRGRG;
Compound 2 (SEQ ID NO. 2):
His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys(Gly-Gly-Ser-Gly-Ser-Gly-γ-Glu-CO(CH 2 ) 18 CO 2 H)-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly;
the abbreviation is: H-Aib-EGTFTSDVSSYLEGQAAK (GGSGSG-gamma-E-CO (CH) 2 ) 18 CO 2 H)EFIAWLVRGRG;
Compound 3 (SEQ ID NO. 3):
His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Glu-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys(Gly-Gly-Ser-Gly-Ser-Gly-γ-Glu-CO(CH 2 ) 18 CO 2 H)-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly;
the abbreviation is: H-Aib-EGTFTSEVSSYLEGQAAK (GGSGSG-gamma-E-CO (CH) 2 ) 18 CO 2 H)EFIAWLVRGRG;
Compound 4 (SEQ ID NO. 4):
His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Glu-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys(Gly-Gly-Ser-Gly-Ser-Gly-γ-Glu-CO(CH 2 ) 18 CO 2 H)-Asp-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly;
the abbreviation is: H-Aib-EGTFTSEVSSYLEGQAAK (GGSGSG-gamma-E-CO (CH) 2 ) 18 CO 2 H)DFIAWLVRGRG;
Compound 5 (SEQ ID NO. 5):
His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys(Gly-Gly-Ser-Gly-Ser-Gly-Ser-Gly-γ-Glu-CO(CH 2 ) 18 CO 2 H)-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly;
the abbreviation is: H-Aib-EGTFTSDVSSYLEGQAAK (GGSGSGSG-gamma-E-CO (CH) 2 ) 18 CO 2 H)EFIAWLVRGRG;
Compound 6 (SEQ ID NO. 6):
His-(D-Ser)-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys(Gly-Gly-Ser-Gly-Ser-Gly-Ser-Gly-γ-Glu-CO(CH 2 ) 18 CO 2 H)-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly;
the abbreviation is: hsEGTFTSDVSSYLEGQAAK (GGSGSGSG-gamma-E-CO (CH) 2 ) 18 CO 2 H)EFIAWLVRGRG;
Compound 7 (SEQ ID NO. 7):
His-(D-Ser)-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys(Gly-Gly-Ser-Gly-Ser-Gly-Ser-Gly-γ-Glu-CO(CH 2 ) 18 CO 2 H)-Gln-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly;
the abbreviation is: hsEGTFTSDVSSYLEGQAAK (GGSGSGSG-gamma-E-CO (CH) 2 ) 18 CO 2 H)QFIAWLVRGRG;
Compound 8 (SEQ ID NO. 8):
His-(D-Ser)-Glu-Gly-Thr-Phe-Thr-Ser-Glu-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys(Gly-Gly-Ser-Gly-Ser-Gly-Ser-Gly-γ-Glu-CO(CH 2 ) 18 CO 2 H)-Asp-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly。
the abbreviation is: hsEGTFTSEVSSYLEGQAAK (GGSGSGSG-gamma-E-CO (CH) 2 ) 18 CO 2 H)DFIAWLVRGRG;
A method for preparing a long-acting GLP-1 polypeptide analog according to an embodiment of the invention comprises the following steps:
step 1: synthesizing a main peptide resin corresponding to a main peptide chain of the long-acting GLP-1 polypeptide analogue according to an Fmoc/t-Bu strategy, wherein the main peptide chain is His-Xaa2-Glu-Gly-Thr-Phe-Thr-Ser-Xaa9-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Xaa21-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly;
and 2, step: on the basis of the main peptide resin, coupling a side arm structure corresponding to the long-acting GLP-1 polypeptide analogue according to an Fmoc/t-Bu strategy to obtain a polypeptide resin corresponding to the long-acting GLP-1 polypeptide analogue; wherein, the structure of the side arm is (Gly) x -(Ser-Gly) y -Z-CO(CH 2 ) n CO 2 H;
And step 3: adding a lysis solution into the polypeptide resin, carrying out a lysis reaction, removing full protection of the polypeptide, and extracting a crude product compound;
purifying the crude compound to obtain the long-acting GLP-1 polypeptide analogue.
According to a method of preparing a GLP-1 polypeptide analogue according to an embodiment of the invention,
in the step 2, coupling agents used are 1-hydroxybenzotriazole and N, N-diisopropylcarbodiimide, a solvent is N, N-dimethylformamide, and Fmoc groups are removed by using 20% piperidine/N, N-dimethylformamide solution;
in step 3, the lysis solution is composed of TFA, DODT, m-cresol and H 2 O is mixed according to the volume ratio of 92.5:2.5:2.5: 2.5; the crude compound extraction means comprises filtration, precipitation and/or methyl tert-butyl ether extraction.
In step 4, the purity of the obtained long-acting GLP-1 polypeptide analogue is more than 96 percent.
It is a further object of the present invention to provide a composition comprising a long-acting GLP-1 polypeptide analogue, further comprising a pharmaceutically acceptable carrier or adjuvant. For example, carriers capable of reducing drug degradation and loss, reducing side effects, such as micelles, microemulsions, gels, and the like; adjuvants refer to materials added to prepare the drug into a suitable dosage form, such as buffers, excipients for lyophilization, etc., and the pharmaceutical composition containing the polypeptide analogs of the present invention can be formulated as a solution or lyophilized powder for parenteral administration, which can be reconstituted with a suitable solvent or other pharmaceutically acceptable carrier prior to use, with liquid formulations typically being buffers, isotonic solutions, and aqueous solutions. The buffer solution can be phosphate buffer solution, the isotonic solution can be 0.9% sodium chloride solution, and the aqueous solution is directly dissolved by purified water.
As will be understood by those skilled in the art, the long-acting GLP-1 polypeptide analogue is used as an active ingredient, and is added with a pharmaceutically acceptable carrier and/or auxiliary material to prepare a pharmaceutical composition, which is suitable for various administration modes, such as oral administration, transdermal administration, intravenous administration, intramuscular administration, local administration, nasal administration and the like. Depending on the mode of administration employed, the pharmaceutical compositions of the polypeptide analogs of the present invention may be formulated in a variety of suitable dosage forms comprising at least one effective amount of the polypeptide analog of the present invention and at least one pharmaceutically acceptable carrier. Examples of suitable dosage forms are tablets, capsules, sugar-coated tablets, granules, oral solutions and syrups, ointments and patches for the skin surface, aerosols, nasal sprays, and sterile solutions for injection.
The amount of the pharmaceutical composition of the present invention may vary over a wide range and can be determined by one skilled in the art based on objective factors such as the type of disease, the severity of the condition, the weight of the patient, the dosage form, the route of administration, and the like.
Still another object of the present invention is to provide the use of the above-mentioned long-acting GLP-1 polypeptide analogues and compositions.
The invention obtains a series of GLP-1 polypeptide analogues, and researches the pharmacodynamic action of the series of medicaments. The GLP-1 receptor agonistic activity, the activity in the aspects of reducing blood sugar, blood fat and weight, treating diabetic nephropathy and the like of the synthesized series of polypeptide analogue medicines are evaluated, and the pharmacokinetics of the polypeptide analogue medicines are preliminarily researched. The result shows that compared with the somaglutide, the long-acting property of the pharmaceutical effect is far better than that of the somaglutide (the half-life period is 2 times or more than that of the somaglutide), and the pharmaceutical effect is also better than that of the somaglutide in treating and improving diabetic nephropathy.
Therefore, the long-acting GLP-1 polypeptide analogue has longer half-life, insulin-promoting activity and no adverse reaction, can be used for treating diabetes and obesity, and can be potentially used as a new generation of medicine for treating diabetes and obesity.
The application of the long-acting GLP-1 polypeptide analogue specifically comprises the following steps:
in the aspect of preparing medicaments for preventing or treating diabetes, in the aspect of preparing medicaments for preventing or treating obesity and in the aspect of preparing health-care products capable of reducing body weight.
The invention has the beneficial effects that:
the solution of polypeptide half-life and stability is the key to the design of polypeptide drugs and the possibility of drug development, and is also a significant scientific and core problem in the field of research. The development of the ultra-long-acting polypeptide drug molecule modification technology is a key and is also a bottleneck to be broken through internationally in the research of the field. According to the invention, through bioinformatics, structural biology, computer-aided design, structure-activity relationship research and the like, a site-specific side arm structure modification technology is developed, an ultra-long-acting polypeptide and protein drug molecule modification neck technology is broken through, the half-life period of a synthesized polypeptide analogue is greatly prolonged, and ultra-long-acting of a polypeptide drug is realized.
The parent peptide in the long-acting GLP-1 polypeptide analogue is a homologous polypeptide, the homologous polypeptide in the invention refers to a polypeptide which originally has the amino acid sequence of glucagon-like peptide (GLP-1) and somaglutide (Semaglutide), but one or more amino acid residues are replaced by different amino acid residues, and a side arm short peptide amino acid residue sequence is added in the structural sequence, the amino acid residues are conserved among each other, and the obtained polypeptide can be used for implementing the invention.
Specifically, we refer to "side arm" short peptide chains linked by amino acid substitutions at specific sites and specific sites described herein (Lys at position 20 in the amino acid sequence), namely: the site-specific side arm modification technology successfully breaks through the neck clamping technology modified by the ultra-long-acting polypeptide drug molecules. Compared with the positive medicine of the somaglutide without side arm modification, the polypeptide medicine designed by the invention greatly prolongs the action half-life period, realizes the ultra-long effect of the polypeptide medicine, and has the integral medicine action effect which is greatly superior to the positive medicine of the somaglutide.
In addition, the GLP-1 polypeptide analogs of the invention utilize lipophilic substituents to bind albumin in the blood, protecting it from enzymatic degradation, thereby increasing half-life. The GLP-1 polypeptide analogs of the invention stabilize the helical structure of the molecule via an intramolecular bridge, improving potency and/or selectivity for the glucagon-like peptide 1 receptor (GLP-1R).
The GLP-1 polypeptide analogue has high synthesis yield, good stability, easy amplification production and low cost.
Compared with the somaglutide, the long-acting property and the stability of the compound having the drug effect are far better than those of the somaglutide (the half-life period of the compound is 2 times or more than that of the somaglutide).
The GLP-1 polypeptide analogue is also superior to the somaglutide in treating and improving diabetes and nephropathy when the mass number is the same.
Meanwhile, the GLP-1 polypeptide analogue has better efficacy of reducing body weight, and the GLP-1 polypeptide analogue can be used for preventing body weight from increasing or promoting body weight from losing by causing food intake reduction and/or energy consumption increase, so that the GLP-1 polypeptide analogue can also be used for directly or indirectly treating other diseases caused by or characterized by overweight, such as treating and/or preventing obesity, morbid obesity, obesity-related inflammation, obesity-related gallbladder disease and sleep apnea caused by obesity, and the effect of the GLP-1 polypeptide analogue in the diseases can be due to the effect of the GLP-1 polypeptide analogue on body weight directly or indirectly or on other aspects of bodies except body weight.
Among the compounds synthesized by the embodiment of the invention, the compounds 2, 5, 6 and 7 have more excellent and more obvious glucose tolerance improving effect, longer drug effect and half-life period compared with the thaumatin, can obviously reduce the liver weight and epididymal fat content of mice, and are also better than the thaumatin in the aspects of improving the oral glucose tolerance (OGTT) and insulin resistance (ITT) of diabetic mice.
Abbreviations used in the present invention have the following specific meanings:
DCM is dichloromethane, DMF is N, N-dimethylformamide, HOBt is 1-hydroxybenzotriazole, fmoc is fluorenylmethoxycarbonyl, resin, FBS is fetal bovine serum, GLP-1R is glucagon-like peptide 1 receptor, GLP-1 is glucagon-like peptide, his is histidine, ser is serine, D-Ser is D-type serine, gln is glutamine, gly is glycine, glu is glutamic acid, ala is alanine, thr is threonine, lys is lysine, arg is arginine, tyr is tyrosine, asp is aspartic acid, trp is tryptophan, phe is phenylalanine, IIe is isoleucine, leu is leucine, cys is cysteine, pro is proline, val is valine, met is methionine, and Asn is asparagine. Aib is 2-aminoisobutyric acid, iva is isovaline.
Drawings
FIG. 1 is a graph of blood glucose monitoring at 24h and 48h after administration based on experimental animals in example 3; wherein:
FIG. 1A is a graph showing the change in blood glucose concentration within 120 minutes of intragastric gavage 24h after administration;
FIG. 1B is the area under the curve (AUC) calculated in FIG. 1A;
FIG. 1C is the change in blood glucose concentration within 120 minutes of intragastric gavage of glucose 48h after dosing;
FIG. 1D is the area under the curve (AUC) calculated in FIG. 1C;
FIG. 2 is a graph of blood glucose monitoring at 72h and 96h after administration based on experimental animals in example 3; wherein:
FIG. 2A is a graph of the change in blood glucose concentration within 120 minutes of intragastric gavage of glucose 72 hours after dosing;
FIG. 2B is the area under the curve (AUC) calculated in FIG. 2A;
FIG. 2C is the change in blood glucose concentration within 120 minutes of intragastric gavage glucose after 96h of administration;
FIG. 2D is the area under the curve (AUC) calculated in FIG. 2C;
FIG. 3 is a graph of body monitoring after administration of 8 week old male db/db diabetic mice in example 4, wherein:
FIG. 3A is a body weight monitor chart of a mouse according to example 4;
FIG. 3B is a graph showing blood glucose monitoring of mice in example 4;
FIG. 3C is a graph showing the monitoring of food intake of the mouse in example 4;
FIG. 4 is a serological index plot of the measurement of material draw after 6 weeks of treatment of 8 week-old db/db diabetic mice, wherein:
FIGS. 4A and 4B are mouse serum ALT and AST results, respectively, with p <0.05;
FIGS. 4C and 4D are mouse serum TG and T-CHO results, respectively, with p <0.05;
FIGS. 4E, 4F and 4G are graphs of mouse serum HDL-C, LDL-C and GHb results, respectively, with p <0.05;
FIG. 5 is a graphical representation of the week 6 physiological index of the mice dosed in example 4, wherein,
FIG. 5A is a graph of mouse liver weight (lever weight);
FIG. 5B is a graph of mouse liver index (lever/body weight);
FIG. 5C is a graph of mouse body weight (body weight);
FIG. 5D is a graph of mouse Body Mass Index (BMI);
FIG. 5E is a graph of epididymal fat weight (EAM weight) in mice;
FIG. 5F is a graph of epididymal fat index (EAM/body weight) in mice;
FIG. 6 is a graph of Oral Glucose Tolerance Test (OGTT) and insulin resistance test (ITT) tests in mice; wherein, the first and the second end of the pipe are connected with each other,
FIG. 6A is a graph of the OGTT results of the mice of example 4 administered at week 4;
FIG. 6B is a histogram of AUC corresponding to FIG. 6A;
FIG. 6C is a graph showing the ITT results at 5 weeks of administration of the mice in example 4;
FIG. 6D is a 5 week histogram of AUC dosing corresponding to FIG. 6C; p <0.05;
fig. 7 is a blood concentration profile of blood collected at different time periods after a single administration in 8-week-old SD rats, in which:
FIGS. 7A1 and 7A2 are the blood concentration curves of SD rat tail vein and subcutaneous injection of Somalutide, respectively;
fig. 7B1 and 7B2 are the blood concentration curves of SD rat tail vein and subcutaneous injection of compound 2, respectively;
fig. 7C1, 7C2 are plasma concentration curves of SD rat tail vein and subcutaneous injection of compound 5, respectively;
fig. 7D1, 7D2 are graphs of the blood concentration of SD rat tail vein and subcutaneous injection of compound 6, respectively;
fig. 7E1, 7E2 are plasma concentration curves of SD rat tail vein and subcutaneous injection of compound 7, respectively;
FIG. 8 shows the molecular formula and structure of fatty acid substituent.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.
Materials:
all amino acids were purchased from jile biochemical, shanghai, somalutide from volume, peptides biology, inc, zhejiang, CAS No.:910463-68-2; all other reagents were analytical grade, purchased from Sigma, unless otherwise specified. Adopts Protein Technologies PRELUDE 6 channel polypeptide synthesizer, phenomenex Luna C 18 A preparative column (46mm. Times.250mm) was used for purifying the polypeptide, a high performance liquid chromatograph was a product of Thermo corporation, and mass spectrometry was performed using a Thermo liquid mass spectrometer.
The material and the method are as follows:
Boc-His (Trt) -OH, fmoc-Aib-OH was purchased from Gele, shanghai, and prepared as mono-tert-butyl eicosanedioate. The remaining amino acids were purchased from Chengdu Zhengyuan, inc., and the condensing agents were purchased from Suzhou Hao Van. All other reagents were analytical grade and solvents were purchased from Shanghai Tatan, unless otherwise specified. The centrifuge was purchased from Luxiang apparatus. 5.0m reverse phase C 18 Preparative columns (46mm X250mm) were used to purify the polypeptides. The high performance liquid chromatograph is a product of Saimerfi company. Mass spectrometry was performed using a Waters mass spectrometer.
EXAMPLE 1 Synthesis of Compound 5
Amino acid sequence of compound 5:
His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys(Gly-Gly-Ser-Glym-γ-Glu-CO(CH 2 ) 18 CO 2 H)-Glu-Phe-Ile-Ala-Trp
-Leu-Val-Arg-Gly-Arg-Gly
the abbreviation is: H-Aib-EGT FTSDV SSYLEGAAK (GGSGSGSG-gamma-E-CO (CH) 2 ) 18 CO 2 H) EFIAW LVRGR G (acetate),
the method comprises the following steps:
Manual synthesis according to Fmoc/t-Bu strategy, scale of synthesis: 0.5mmol, the following main peptide resins were synthesized:
Boc-His-Aib-Glu(OtBu)-Gly-Thr(tBu)-Phe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Val-Ser(tBu)-Ser(tBu)-Tyr(tBu)-Leu-Glu(OtBu)-Gly-Gln(Trt)-Ala-Ala-Lys(Alloc)-Glu(OtBu)-Phe-Ile-Ala-Trp(Boc)-Leu-Val-Arg(pbf)-Gly-Arg(pbf)-Gly-Wang Resin。
(1): weighing 1.47 g Wang Resin (loading 0.47mmol/g, xian Langxi), adding into a reaction column, adding into N, N-Dimethylformamide (DMF), swelling for 30min, weighing Fmoc-Gly-OH:1.233g (6 eq), HOBT:0.672g (7.2 eq), DMAP:0.06g (0.72 eq) for use. The DMF was taken off, the resin was washed thoroughly with N, N-Dimethylformamide (DMF) 2 times, and the weighed material was added to the reaction column. An appropriate amount of DMF was added, nitrogen was added thereto and the mixture was stirred well, followed by addition of 0.83mL (7.8 eq) of N, N-Diisopropylcarbodiimide (DIC). And (5) reacting for 2h, and finishing the reaction. The reaction was taken off, washed 3 times with DMF and blocked for 4h by addition of acetic anhydride/pyridine (7,v/v). And (4) pumping the confining liquid, and washing 6 times by using DMF to obtain Fmoc-Gly-Wang Resin.
(2): fmoc-Gly-Wang Resin is used as a carrier, 1-Hydroxybenzotriazole (HOBT) and N, N-Diisopropylcarbodiimide (DIC) are used as coupling agents, N, N-Dimethylformamide (DMF) is used as a solvent, an Fmoc group is removed by a 20% Piperidine (Piperidine)/N, N-Dimethylformamide (DMF) solution (two times, 5min +7 min), and the coupling effect is monitored by ninhydrin in the coupling process. By performing manual feeding, fmoc-Arg (pbf) -OH, fmoc-Gly-OH, fmoc-Arg (pbf) -OH, fmoc-Val-OH, fmoc-Leu-OH, fmoc-Trp (Boc) -OH, fmoc-Ala-OH, fmoc-Ile-OH, fmoc-Phe-OH, fmoc-Glu (OtBu) -OH, fmoc-Lys (Alloc) -OH, fmoc-Ala-OH, fmoc-Ala-OH, fmoc-Ala-OH, fmoc-Gln (Trt) -OH, fmoc-Gly-OH, fmoc-Glu (OtBu) -OH, fmoc-Leu-OH, fmoc-Tyr (tBu) -OH, fmoc-Ser (tBu) -OH were connected by condensation reaction sequentially from C-terminus to N-terminus, fmoc-Ser (tBu) -OH, fmoc-Val-OH, fmoc-Asp (OtBu) -OH, fmoc-Ser (tBu) -OH, fmoc-Thr (tBu) -OH, fmoc-Phe-OH, fmoc-Thr (tBu) -OH, fmoc-Gly-OH, fmoc-Glu (OtBu) -OH, fmoc-Aib-OH, boc-His (Trt) -OH the above amino acids were dosed (5 eq relative to the synthesis scale) to give Boc-His-Aib-Glu (OtBu) -Gly-Thr (tBu) -Phe-Thr (tBu) -Ser (tBu) -Asp (OtBu) -Val-Ser (tBu) -Ser (tBu) -Tyr (tBu) - Leu-Glu (OtBu) -Gly-Gln (Trt) -Ala-Ala-Lys (Alloc) -Glu (OtBu) -Phe-Ile-Ala-Trp (Boc) -Leu-Val-Arg (pbf) -Gly-Arg (pbf) -Gly-Wang Resin.
There are several points to be explained:
1) The synthesis of Fmoc-Gly-Wang Resin has the disadvantages that the substitution degree of Wang Resin is low, the feeding amount of Fmoc-Gly-OH is large, otherwise, the substitution degree is low, and the material is wasted. Blocking with acetic anhydride/pyridine prevented the appearance of defective peptides.
2) In the following condensation reaction, the inventory of Fmoc protected amino acid, 1-Hydroxybenzotriazole (HOBT) and N, N-Diisopropylcarbodiimide (DIC) is 5 times, and the reaction time is 2 hours.
(3): removal of allyloxycarbonyl (Alloc)
Dichloromethane (DCM) was added to the resin, morpholine 0.5mL (12 eq) was added, and 0.173g of palladium tetratriphenylphosphine (0.3 eq) was weighed into the reaction column and reacted for 1h. After the reaction, the reaction solution was removed by suction, washed 3 times with N, N-Dimethylformamide (DMF) and 6 times with DCM. Obtaining a main peptide resin: boc-His-Aib-Glu (OtBu) -Gly-Thr (tBu) -Phe-Thr (tBu) -Ser (tBu) -Asp (OtBu) -Val-Ser (tBu) -Ser (tBu) -Tyr (tBu) -Leu-Glu (OtBu) -Gly-Gln (Trt) -Ala-Ala-Lys-Glu (OtBu) -Phe-Ile-Ala-Trp (Boc) -Leu-Val-Arg (pbf) -Gly-Arg (pbf) -Gly-Wang Resin.
coupling Fmoc-Gly-OH according to Fmoc/t-Bu strategy: adding Fmoc-Gly-OH, HOBT and a proper amount of N, N-Dimethylformamide (DMF) into the main peptide resin product, uniformly stirring with nitrogen, adding DIC, stirring with nitrogen for reaction for 2 hours, detecting the coupling effect with ninhydrin, and detecting the reaction to obtain the product with colorless transparency. And (3) pumping out reaction liquid, washing with N, N-Dimethylformamide (DMF) for 3 times, removing Fmoc groups (two times of 5min + 7min) by using 20% Piperidine (Piperidine)/N, N-Dimethylformamide (DMF) solution, washing with DMF for 6 times after removing Fmoc, sampling ninhydrin for detection, and developing color.
The operations are repeated to couple Fmoc-Gly-OH, fmoc-Ser (tBu) -OH, fmoc-Gly-OH, fmoc-Ser (tBu) -OH, fmoc-Gly-OH, fmoc-Ser (tBu) -OH, fmoc-Gly-OH, fmoc-Glu-OtBu and mono-tert-butyl didecylate in sequence. Obtaining Boc-His-Aib-Glu (OtBu) -Gly-Thr (tBu) -Phe-Thr (tBu) -Ser (tBu) -Asp (OtBu) -Val-Ser (tBu) -Ser (tBu) -Tyr (tBu) -Leu-Glu (OtBu) -Gly-Gln (Trt) -Ala-Lys (Gly-Gly-Ser (tBu) Gly-Ser (tBu) -Gly-Ser (tBu) -Gly-Glu-OtBu-ditert-butyl didecanedioate) -Glu (OtBu) -Phe-Ile-Ala-Trp (Boc) -Leu-Val-Arg (pbf) -Gly-Arg (pbf) -Gly-Wang Resin. Washing with N, N-Dimethylformamide (DMF) for 3 times, washing with Dichloromethane (DCM) for 3 times, shrinking with Methanol (Methanol) for 2 times, and vacuum drying to obtain dried polypeptide resin.
Lysis solution: TFA, DODT, m-cresol, H 2 O is mixed according to the volume ratio of 92.5:2.5:2.5: the mixture is prepared in advance according to the proportion of 2.5, and is frozen in a refrigerator for 2 hours.
To the dried polypeptide Resin Boc-His-Aib-Glu (OtBu) -Gly-Thr (tBu) -Phe-Thr (tBu) -Ser (tBu) -Asp (OtBu) -Val-Ser (tBu) -Ser (tBu) -Tyr (tBu) -Leu-Glu (OtBu) -Gly-Gln (Trt) -Ala-Ala-Lys (Gly-Gly-Ser (tBu) -Gly-Ser (tBu) -Gly-Ser (tBu) -Gly-Glu-OtBu-didecanedioic acid mono-tert-butyl ester) -Glu (OtBu) -Phe-Ile-Ala-Trp (Boc) -Leu-Val-Arg (pbf) -Gly-Arg (pbf) -Gly-Wang Resin was added lysate per 10mL of lysate/g polypeptide Resin, the lysate was warmed to room temperature, and the cleavage reaction was carried out for 3 hours. Filtering, washing the filter cake with a small amount of lysate for 3 times, and combining the filtrates. The filtrate was slowly poured into ice methyl tert-butyl ether with stirring. Standing for more than 2 hours until the precipitate is complete. The supernatant was removed, the precipitate was centrifuged, washed 3 times with methyl tert-butyl ether, centrifuged and the solid was blown dry with nitrogen. To obtain a crude compound His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys (Gly-Gly-Ser-Gly-Ser-Gly-Glu-eicosanedioic acid) -Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly.
The crude compound obtained in step 2 was dissolved in Acetonitrile (ACN): h 2 O =1:2 (volume ratio), passing 5.0m of reverse phase C 18 Preparative HPLC purification was performed on a packed 46mm x 250mm column. With 45% acetonitrile/H 2 Starting with O (1% trifluoroacetic acid), the column was eluted with a gradient (increasing acetonitrile ratio at 0.33%/min) at a flow rate of 10mL/min for 6At 0min, fractions containing the polypeptide were collected, giving samples with HPLC purity greater than 90%. HPLC purification was repeated once starting with 31% acetonitrile/20 mM sodium dihydrogen phosphate in water and pH adjusted to 6.5 with 1M sodium hydroxide solution, using a gradient (0.33%/min increasing acetonitrile ratio) at a flow rate of 10mL/min, eluting the column for 60 minutes, collecting the polypeptide-containing fractions, and freeze-drying to obtain a purity of greater than 96.86% with a total yield of 15%.
Identifying the separated product polypeptide by liquid chromatography-mass spectrometry, and finding that the m/z value of the ion peak of the protonated molecule is as follows: 4397.16, target compound 5, theoretical compound 5 molecular weight 4397.87.
Synthesis of Compound 2
Since compound 2 differs from compound 5 by the sequence of the "side-arm" short peptide chain, the synthetic steps of compound 2 differ from compound 5 by step 2 coupling of the "side-arm" structure:
in the step, fmoc-Gly-OH, fmoc-Ser (tBu) -OH, fmoc-Gly-OH, fmoc-Ser (tBu) -OH, fmoc-Gly-OH, fmoc-Glu-OtBu and mono-tert-butyl didecanedioate are sequentially coupled. Obtaining Boc-His-Aib-Glu (OtBu) -Gly-Thr (tBu) -Phe-Thr (tBu) -Ser (tBu) -Asp (OtBu) -Val-Ser (tBu) -Ser (tBu) -Tyr (tBu) -Leu-Glu (OtBu) -Gly-Gln (Trt) -Ala-Ala-Lys (Gly-Gly-Ser (tBu) Gly-Ser (tBu) -Gly-Glu-OtBu-undecanedioic acid mono-tert-butyl ester) -Glu (OtBu) -Phe-Ile-Ala-Trp (Boc) -Leu-Val-Arg (pbf) -Gly-Wang Resin.
The purification of compound 2 and the confirmation of the product were carried out in the same manner as in compound 5, and the purity was more than 97.66%, and the total yield was 12%. The separated product was identified by LC-MS and the m/z values of the ion peaks of the protonated molecules were found to be: 4253.07 is target compound 2, and the theoretical molecular weight of compound 2 is 4253.74.
Based on the above synthesis steps, purification and product confirmation methods, the coupling order of the amino acids in the compound synthesis steps 1 and 2 was adjusted according to the difference in amino acids in the main peptide chain and the side arm "short peptide chain" of the compounds 1, 3, 4, 6, 7, 8, and finally the corresponding target product was synthesized, and the separated product was identified by liquid chromatography-mass spectrometry, the m/z value of the protonated molecular ion peak (the actual value in table 1) was confirmed, and the actual value was compared with the theoretical value of molecular weight to confirm that the synthesized and purified product was the target product, and the theoretical value of molecular weight, the actual value of liquid chromatography-mass spectrometry, the sequence and the molecular formula of the compounds 1 to 8 were given in table 1, respectively.
TABLE 1 Long-acting GLP-1 polypeptide analogue amino acid sequence table and liquid chromatography-mass spectrometry combined identification result
EXAMPLE 2 evaluation of agonistic Activity of Compounds 1-8 and Somaglutide on GLP-1R (glucagon-like peptide 1 receptor)
The agonistic activity of GLP-1R was determined for compounds 1-8 and somaglutide in a GLP-1R-Luciferase-HEK293 cell model (cell line constructed in this laboratory).
First, cells were digested in 96-well plates (medium containing 10% FBS, GLP-1R-Luciferase-HEK293:20000cell/well, 100. Mu.L); after 36h, the culture medium in the 96-well plate is discarded, and 90 mu L of serum-free culture medium is added; after 6h, serial concentrations (0.01, 0.1, 1, 10, 100, 1000, 10000, 100000, 1000000, 10000000, 100000000 pM) of polypeptide drugs (8 polypeptide analogs and somaglutide) were prepared in serum-free medium, 10 μ L (i.e. diluted 10-fold) was added to each well, and the cells were incubated for 5h; adding 100 mu L of cell lysate into each well, cracking on ice for 10min, then shaking uniformly, taking 2 mu L of lysate, adding into a white board of a 384 enzyme-labeling instrument, adding 10 mu L of firefly luciferase reaction solution, reading, adding 10 mu L of renilla luciferase reaction solution, and reading; data processing, the number of the firefly luciferase reading is divided by the Renilla luciferase reading, and the blank value is subtracted to obtain the activation times at different concentrations.
TABLE 2 EC of Compounds 1-8 with Somuramyl peptide agonizing GLP-1R 50
EC 50 | |
Somazutide | 222.2 |
Compound | |
1 | 699.1 |
Compound | |
2 | 64.73 |
Compound | |
3 | 1003.7 |
Compound | |
4 | 1349.9 |
Compound | |
5 | 146.3 |
Compound | |
6 | 203.5 |
Compound | |
7 | 302.4 |
Compound | |
8 | 1599.2pM |
From experimental knotAs can be seen, compounds 1-8 of the present invention all activated GLP-1R, and compounds 2, 5, 6 and 7 had better agonistic activity against GLP-1R (Table 2), wherein half maximal effect concentration (hereinafter abbreviated as EC) of compounds 2, 5 50 That is, a concentration that causes 50% of the maximal effect) is lower than thaumareuptade, indicating that the agonistic activity towards GLP-1R is superior to thaumareuptade, while the agonistic activity towards GLP-1R of compounds 6, 7 is comparable to thaumareuptade.
EXAMPLE 3 Effect of Compounds 1-8 and Somalitide on oral glucose tolerance (OGTT)
Male C57BL/6J mice (Experimental animals center, zhongshan university) of around 8 weeks of age were housed for one week in acclimation and randomly grouped into groups of 8 per group according to similar blood glucose (evaluation of blood samples taken from the tail tip).
The compounds 1-8 of the present invention (corresponding to numbers 1-8 in the figures) and somaglutide (corresponding to semaglutide in the figures) were administered subcutaneously at a dose of 120ug/kg, and the control group was administered with the same dose of PBS. ddH for intragastric glucose 2 O is prepared into stock solution with the concentration of 0.5g/mL and is stored at normal temperature. Mice were fasted for 8h before gavage, gavage glucose at a dose of 2.5g/kg at 24h, 48h, 72h and 96h post-dose, respectively, and blood glucose was measured at t =0min, t =15min, t =30min, t =60min, t =90min and t =120 min. The data were processed using software GraphPadPrism to generate a line graph of blood glucose changes and calculate the area under the curve to generate an AUC plot, the results of which are shown in fig. 1 and fig. 2.
According to the AUC (area under concentration-time curve) counted by the OGTT curve, the bioavailability is high when the AUC is large, and the bioavailability is low when the AUC is not large. Wherein in fig. 1 and 2: p <0.05; * *: p <0.01; * **: representing p <0.001.
As can be seen from fig. 1A and B, the sumatriptan and compounds 1-8 significantly reduced AUC compared to vehicle (PBS) after 24h administration, indicating that sumatriptan and compounds 1-8 have significant glucose tolerance.
As can be seen from fig. 1C, D and fig. 2, the effect of somaglutide on AUC did not differ significantly from vehicle (PBS) after 48h, 72h, 96h of administration, indicating that somaglutide has failed to improve glucose tolerance. After 48h, 72h, 96h of administration, compounds 2, 5, 6 and 7 still significantly reduced AUC compared to PBS group, and thus improved glucose tolerance, and thus, compounds 2, 5, 6 and 7 were all superior to somaglutide in improving glucose tolerance in mice.
As can be seen from the above, compounds 1 to 8 of the present invention all have the effect of improving glucose tolerance in mice to different degrees compared to vehicle (PBS), wherein compounds 2, 5, 6 and 7 exhibit more excellent and significant glucose tolerance improvement effect in 4 OGTT curve periods (24, 48, 72, 96 h), and the results also indicate that compounds 1 to 8, especially compounds 2, 5, 6 and 7 exhibit more excellent and significant long-acting glucose lowering effect in 4 OGTT curve periods (24, 48, 72, 96 h) relative to thaumautide.
EXAMPLE 4 therapeutic Effect of Compounds 2, 5, 6, 7 and Somalutide on diabetes in diabetic mice
48 male Lepr db/db mice (db/db) at 8 weeks of age and 8 littermate normal mice (WT) were purchased from Nanjing university-Nanjing biomedical research institute.
A db/db mouse diabetes model was obtained (purchased from Nanjing university-Nanjing biomedical research institute, mice about 8 weeks, and blood glucose and body weight were measured to ensure smooth performance of subsequent experiments), db/db mice were randomly divided into 6 groups according to blood glucose ( compounds 2, 5, 6, 7 and somatid, physiological saline group), each group had 8 mice, and there was no difference in basal body weight and blood glucose. Each group of mice was injected subcutaneously every other day with compounds 2, 5, 6, 7 (120. Mu.g/kg), somaglutide (120. Mu.g/kg) and saline (WT group and db/db group), respectively. After each administration of the mice, fasting for 6h every other day to measure the blood sugar and body weight of the mice; the water intake and food intake were measured every 7 days. OGTT was measured at week 4 of dosing, insulin tolerance (ITT) at week 5, and samples taken at week 6, and various serological and physiological indices were examined. The monitoring results of blood sugar, body weight, water intake and food intake of the mice are shown in figure 3, the serological indexes after material taking are shown in figure 4, and the physiological indexes are shown in figure 5.
A model of type 2 diabetes characterized by: obesity, insulin resistance, hyperglycemia, dyslipidemia, liver steatosis, and the like. Oral Glucose Tolerance Test (OGTT) and insulin resistance test (ITT) were performed at 4 and 5 weeks of treatment of diabetic mice with compounds 2, 5, 6, 7 and somaglutide, respectively, and the results are shown in fig. 6.
FIG. 3 is a graph of post-administration body monitoring of medium 8 week old male db/db diabetic mice, wherein normal saline, compound 2 (120. Mu.g/kg), compound 5 (120. Mu.g/kg), compound 6 (120. Mu.g/kg), compound 7 (120. Mu.g/kg) and Somalutide (120. Mu.g/kg) were injected subcutaneously for 6 weeks, respectively; body weight and blood glucose of mice were measured every 3 days after 6h fasting (3A and 3B), and food intake of mice was measured every 7 days (3C), wherein: p <0.05; * *: represents p <0.01; * **: representing p <0.001.
The results in fig. 3 (a, C) show that compounds 2, 5, 6, and 7 significantly reduced the food intake of mice compared to db/db group, thereby reducing the weight of mice, and the weight reduction effect was superior to thaumatin.
The results in fig. 3B show that compounds 2, 5, 6, 7 and the positive control drug somaglutide significantly reduced fasting plasma glucose in db/db mice compared to db/db group, indicating that these drugs had significant hypoglycemic effects, blood glucose had decreased to normal levels by week 1 and thereafter blood glucose was relatively stable and superior to somaglutide.
Fig. 4 (a, B) shows that compounds 2, 5, 6 and 7 are also comparable to somaglutide in terms of liver protection function.
Glycated hemoglobin (GHb) can be used as a control index reflecting that the diabetic obtains blood sugar for a long period of time (4-10 weeks), the glycated hemoglobin is increased due to the long-term poor control of the blood sugar, so the glycated hemoglobin measurement is helpful for control, and has an important role in the study of peripheral vascular and cardiovascular complications of diabetes, and the experimental results in fig. 4G show that compounds 2, 5, 6 and 7 have a significant effect of reducing glycated hemoglobin and are superior to thaumatin.
The results in fig. 5E show that compounds 2, 5, 6 and 7 can significantly reduce the liver weight and epididymal fat content of mice, and are superior to thaumatin.
The results in fig. 6 show that compounds 2, 5, 6 and 7 are superior to the positive control drug somaglutide in improving oral glucose tolerance (OGTT) and insulin resistance (ITT) in diabetic mice, as seen in the results.
Example 5 pharmacokinetic study of Compounds 2, 5, 6 and 7 and Somaloutide in SD rats
SD rats (Experimental animal center of Zhongshan university) of about 8 weeks old are bred for one week to adapt to the environment, and the male and female animals are randomly grouped according to the body weight, and 6 animals (male and female animals) are grouped in each group for 5 groups.
In this example, compounds 2, 5, 6 and 7 and somaglutide were administered at a dose of 50ug/kg, a volume of 2mL/kg, a concentration of 60ug/mL, and a vehicle of physiological saline, subcutaneously and in the tail vein, respectively.
The rats in the single tail vein administration group had blood samples collected through jugular veins before (0 min) and 5min, 15min, 30min, 1h, 2h, 4h, 6h, 8h, 24h, 36h, 48h, 72h, 4d, 5d, 6d, 7d, 8d, 9d, 10d after administration, and the collection amount was about 0.2mL at each time point. The rats in the single subcutaneous injection group had blood samples collected via jugular vein before (0 min) and 10min, 20min, 30min, 1h, 2h, 4h, 6h, 8h, 24h, 36h, 48h, 72h, 4d, 5d, 6d, 7d, 8d, 9d, 10d after administration, with a collection volume of about 0.2mL at each time point. Placing in a container containing anticoagulant K 2 EDTA in EP tubes, and centrifugation (3500 g, 10min) at 2-8 ℃ within 1 hour after blood collection, and the plasma separated after centrifugation was filled in labeled EP tubes, and the drug concentrations of compounds 2, 5, 6, 7 and Somalutide in the plasma were determined by LC-MS/MS analysis. Data processing pharmacokinetic parameters were calculated using a non-compartmental model in WinNonlin 6.4 software.
As can be seen from the results of the experiment in FIG. 7, after a single subcutaneous injection of compounds 2, 5, 6 and 7 (50. Mu.g/kg), the absorption in rats was slow and the time to peak T was observed max All are about 24h, half-life period t 1/2 On average 14.99h, 15.4h, 16.6h and 15.42h, respectively, compounds 2, 5, 6 and 7 were all higher than thaumatin (7.87 h). Half-life t after a single intravenous injection of Compounds 2, 5, 6 and 7 (50. Mu.g/kg) 1/2 The average time is 14.49h, 15.2h, 16.2h and 15.1h respectively, and the average time is superior to that of the somaglutide (the average time is 8 h).
The above description is only for the specific embodiments of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art can easily think of the changes or substitutions within the technical scope of the present invention, and shall cover the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Sequence listing
<110> Qingdao Borui scientific & technological Limited
<120> long-acting GLP-1 polypeptide analogue, preparation method and application thereof
<141> 2021-07-19
<160> 8
<170> SIPOSequenceListing 1.0
<210> 1
<211> 31
<212> PRT
<213> Artificial Synthesis ()
<220>
<221> MISC_FEATURE
<222> (2)..(2)
<223> Xaa=Aib
<220>
<221> UNSURE
<222> (2)..(2)
<223> The 'Xaa' at location 2 stands for Gln, Arg, Pro, or Leu.
<400> 1
His Xaa 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 Arg Gly Arg Gly
20 25 30
<210> 2
<211> 31
<212> PRT
<213> Artificial Synthesis ()
<220>
<221> MISC_FEATURE
<222> (2)..(2)
<223> Xaa=Aib
<220>
<221> UNSURE
<222> (2)..(2)
<223> The 'Xaa' at location 2 stands for Gln, Arg, Pro, or Leu.
<400> 2
His Xaa 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 Arg Gly Arg Gly
20 25 30
<210> 3
<211> 31
<212> PRT
<213> Artificial Synthesis ()
<220>
<221> MISC_FEATURE
<222> (2)..(2)
<223> Xaa=Aib
<220>
<221> UNSURE
<222> (2)..(2)
<223> The 'Xaa' at location 2 stands for Gln, Arg, Pro, or Leu.
<400> 3
His Xaa Glu Gly Thr Phe Thr Ser Glu Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Arg Gly Arg Gly
20 25 30
<210> 4
<211> 31
<212> PRT
<213> Artificial Synthesis ()
<220>
<221> MISC_FEATURE
<222> (2)..(2)
<223> Xaa=Aib
<220>
<221> UNSURE
<222> (2)..(2)
<223> The 'Xaa' at location 2 stands for Gln, Arg, Pro, or Leu.
<400> 4
His Xaa Glu Gly Thr Phe Thr Ser Glu Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Asp Phe Ile Ala Trp Leu Val Arg Gly Arg Gly
20 25 30
<210> 5
<211> 31
<212> PRT
<213> Artificial Synthesis ()
<220>
<221> MISC_FEATURE
<222> (2)..(2)
<223> Xaa=Aib
<220>
<221> UNSURE
<222> (2)..(2)
<223> The 'Xaa' at location 2 stands for Gln, Arg, Pro, or Leu.
<400> 5
His Xaa 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 Arg Gly Arg Gly
20 25 30
<210> 6
<211> 31
<212> PRT
<213> Artificial Synthesis ()
<220>
<221> MISC_FEATURE
<222> (2)..(2)
<223> D-Ser
<400> 6
His Ser 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 Arg Gly Arg Gly
20 25 30
<210> 7
<211> 31
<212> PRT
<213> Artificial Synthesis ()
<220>
<221> MISC_FEATURE
<222> (2)..(2)
<223> D-Ser
<400> 7
His Ser Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Gln Phe Ile Ala Trp Leu Val Arg Gly Arg Gly
20 25 30
<210> 8
<211> 31
<212> PRT
<213> Artificial Synthesis ()
<220>
<221> MISC_FEATURE
<222> (2)..(2)
<223> D-Ser
<400> 8
His Ser Glu Gly Thr Phe Thr Ser Glu Val Ser Ser Tyr Leu Glu Gly
1 5 10 15
Gln Ala Ala Lys Asp Phe Ile Ala Trp Leu Val Arg Gly Arg Gly
20 25 30
Claims (5)
1. A long-acting GLP-1 polypeptide analog, wherein the amino acid sequence of said long-acting GLP-1 polypeptide analog is as follows: his-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys (Gly-Gly-Ser-Gly-Ser-Gly- γ -Glu-CO (CH) 2 ) 18 CO 2 H)-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly。
2. A method for preparing a long-acting GLP-1 polypeptide analogue according to claim 1, comprising the steps of:
step 1: synthesizing a main peptide resin corresponding to a main peptide chain of the long-acting GLP-1 polypeptide analogue according to the Fmoc/t-Bu strategy, wherein the main peptide chain is His-Aib-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-Arg-Gly-Arg-Gly;
step 2: on the basis of the main peptide resin, coupling a side arm structure corresponding to the long-acting GLP-1 polypeptide analogue according to an Fmoc/t-Bu strategy to obtain a polypeptide resin corresponding to the long-acting GLP-1 polypeptide analogue; wherein the structure of the side arm is Gly-Gly-Ser-Gly-Ser-Gly-Ser-Gly-gamma-Glu-CO (CH) 2 ) 18 CO 2 H;
And step 3: adding a lysis solution into the polypeptide resin, carrying out a cleavage reaction, removing full protection of the polypeptide, and extracting a crude product compound;
and 4, step 4: purifying the crude compound to obtain the long-acting GLP-1 polypeptide analogue.
3. The method for producing a long-acting GLP-1 polypeptide analog according to claim 2,
in the step 2, coupling agents used are 1-hydroxybenzotriazole and N, N-diisopropylcarbodiimide, a solvent is N, N-dimethylformamide, and Fmoc groups are removed by using 20% piperidine/N, N-dimethylformamide solution;
in step 3, the lysis solution is composed of TFA, DODT, m-cresol and H 2 O is mixed according to a volume ratio of 92.5:2.5:2.5: 2.5; the crude compound extraction means comprises filtration, precipitation and/or methyl tert-butyl ether extraction.
4. A composition comprising a long-acting GLP-1 polypeptide analogue of claim 1 and a pharmaceutically acceptable carrier or adjuvant.
5. Use of a long-acting GLP-1 polypeptide analogue of claim 1 for the preparation of a medicament for the treatment of diabetes;
alternatively, the long-acting GLP-1 polypeptide analogue is used for preparing a medicament for treating obesity.
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110815583.3A CN113429471B (en) | 2021-07-19 | 2021-07-19 | Long-acting GLP-1 polypeptide analogue, and preparation method and application thereof |
CN202211555764.8A CN116554299A (en) | 2021-07-19 | 2021-07-19 | Long-acting GLP-1 polypeptide analogue, and preparation method and application thereof |
CN202211555750.6A CN116410297A (en) | 2021-07-19 | 2021-07-19 | Long-acting GLP-1 polypeptide analogue, and preparation method and application thereof |
PCT/CN2021/107770 WO2023000240A1 (en) | 2021-07-19 | 2021-07-22 | Long-acting glp-1 polypeptide analogue, and preparation method therefor and use thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110815583.3A CN113429471B (en) | 2021-07-19 | 2021-07-19 | Long-acting GLP-1 polypeptide analogue, and preparation method and application thereof |
Related Child Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202211555750.6A Division CN116410297A (en) | 2021-07-19 | 2021-07-19 | Long-acting GLP-1 polypeptide analogue, and preparation method and application thereof |
CN202211555764.8A Division CN116554299A (en) | 2021-07-19 | 2021-07-19 | Long-acting GLP-1 polypeptide analogue, and preparation method and application thereof |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113429471A CN113429471A (en) | 2021-09-24 |
CN113429471B true CN113429471B (en) | 2022-12-23 |
Family
ID=77761113
Family Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202211555750.6A Pending CN116410297A (en) | 2021-07-19 | 2021-07-19 | Long-acting GLP-1 polypeptide analogue, and preparation method and application thereof |
CN202110815583.3A Active CN113429471B (en) | 2021-07-19 | 2021-07-19 | Long-acting GLP-1 polypeptide analogue, and preparation method and application thereof |
CN202211555764.8A Pending CN116554299A (en) | 2021-07-19 | 2021-07-19 | Long-acting GLP-1 polypeptide analogue, and preparation method and application thereof |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202211555750.6A Pending CN116410297A (en) | 2021-07-19 | 2021-07-19 | Long-acting GLP-1 polypeptide analogue, and preparation method and application thereof |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202211555764.8A Pending CN116554299A (en) | 2021-07-19 | 2021-07-19 | Long-acting GLP-1 polypeptide analogue, and preparation method and application thereof |
Country Status (2)
Country | Link |
---|---|
CN (3) | CN116410297A (en) |
WO (1) | WO2023000240A1 (en) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114716533B (en) * | 2022-04-12 | 2023-04-14 | 北京惠之衡生物科技有限公司 | Acylated long-acting GLP-1 derivative |
CN115947822B (en) * | 2022-07-04 | 2023-08-18 | 北京惠之衡生物科技有限公司 | Long-acting acylated insulin derivative, and pharmaceutical composition and application thereof |
CN117756913A (en) * | 2022-11-07 | 2024-03-26 | 内蒙古博睿精创科技有限公司 | Novel long-acting polypeptide compound, composition and application thereof |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
TWI362392B (en) * | 2005-03-18 | 2012-04-21 | Novo Nordisk As | Acylated glp-1 compounds |
US8614182B2 (en) * | 2009-07-30 | 2013-12-24 | Jiangsu Hansoh Pharmaceuticals Co., Ltd. | GLP-1 analogues and their pharmaceutical salts and uses |
WO2013029279A1 (en) * | 2011-09-03 | 2013-03-07 | 深圳市健元医药科技有限公司 | Novel glp-i analogue, preparation method and use thereof |
AR104932A1 (en) * | 2015-06-22 | 2017-08-23 | Lilly Co Eli | GLUCAGON AND PEPTIDE-1 CO-AGONIST COMPOUNDS SIMILAR TO GLUGACON (GLP-1) |
US11130794B2 (en) * | 2017-07-19 | 2021-09-28 | Novo Nordisk A/S | Bifunctional compounds |
CN111253475B (en) * | 2020-02-18 | 2021-03-09 | 江苏诺泰澳赛诺生物制药股份有限公司 | GLP-1 agonist polypeptide compound and salt thereof, and synthesis method and application thereof |
CN112110981B (en) * | 2020-09-23 | 2021-06-25 | 深圳深创生物药业有限公司 | Preparation method of polypeptide containing long-chain fatty diacid side chain |
CN112898405B (en) * | 2021-01-22 | 2023-02-28 | 深圳市图微安创科技开发有限公司 | Polypeptide compound and application thereof in preventing or treating diabetes or diabetic complications |
-
2021
- 2021-07-19 CN CN202211555750.6A patent/CN116410297A/en active Pending
- 2021-07-19 CN CN202110815583.3A patent/CN113429471B/en active Active
- 2021-07-19 CN CN202211555764.8A patent/CN116554299A/en active Pending
- 2021-07-22 WO PCT/CN2021/107770 patent/WO2023000240A1/en unknown
Also Published As
Publication number | Publication date |
---|---|
CN116554299A (en) | 2023-08-08 |
WO2023000240A1 (en) | 2023-01-26 |
CN116410297A (en) | 2023-07-11 |
CN113429471A (en) | 2021-09-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
AU2015321161B2 (en) | Oxyntomodulin analogue | |
CN113429471B (en) | Long-acting GLP-1 polypeptide analogue, and preparation method and application thereof | |
CN112409460B (en) | GLP-1/glucagon receptor dual agonist and application thereof | |
CN111253475B (en) | GLP-1 agonist polypeptide compound and salt thereof, and synthesis method and application thereof | |
CN106928341B (en) | Fixed-point mono-substituted pegylated Exendin analogue and preparation method thereof | |
KR20040070237A (en) | Insulin Molecule Having Protracted Time Action | |
CN106661096B (en) | Novel exenatide analogue and application thereof | |
CN112898406B (en) | GLP-1 analogue peptide modified dimer with different configurations and application of preparation method thereof in treatment of type II diabetes | |
AU2024203093A1 (en) | Polypeptide compounds and use thereof in the prevention or treatment of diabetes or diagnostic compounds | |
CN105968186B (en) | Glucagon (Glu) analogue with long-acting effect and application thereof | |
KR20230008846A (en) | Polypeptide derivatives with dual receptor agonism and their uses | |
CN110759991B (en) | Gemfibrozil-xenopus laevis glucagon-like peptide-1 derivative and application thereof | |
CN113754752B (en) | Polypeptide and preparation method and application thereof | |
CN116514952B (en) | GLP-1 analogues and application thereof | |
CN115232200B (en) | Long-acting Exendin-4 analogue and application thereof | |
CN117756913A (en) | Novel long-acting polypeptide compound, composition and application thereof | |
CN115073582A (en) | Polypeptide compound and application thereof in preventing or treating diabetes or diabetic complications |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant | ||
CP03 | Change of name, title or address | ||
CP03 | Change of name, title or address |
Address after: 017299 1701, Floor 17, Dongfang Building, Altan Xire, Ejin Horo Banner Banner, Ordos City, Inner Mongolia Patentee after: Inner Mongolia Borui Jingchuang Technology Co.,Ltd. Address before: 266600 Room 101, No. 14, Zhenxing Road, Nanshu Town, Laixi City, Qingdao City, Shandong Province Patentee before: Qingdao Borui Jingchuang Technology Co.,Ltd. |