WO2021143810A1 - Composé polypeptidique et son utilisation - Google Patents

Composé polypeptidique et son utilisation Download PDF

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WO2021143810A1
WO2021143810A1 PCT/CN2021/072049 CN2021072049W WO2021143810A1 WO 2021143810 A1 WO2021143810 A1 WO 2021143810A1 CN 2021072049 W CN2021072049 W CN 2021072049W WO 2021143810 A1 WO2021143810 A1 WO 2021143810A1
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γglu
compound
aeeac
seq
aib
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PCT/CN2021/072049
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Chinese (zh)
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黄亮
曹春来
邓慧兴
周翠
刘合栋
谢鑫
何秀仪
杨晓纯
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珠海联邦制药股份有限公司
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Priority to CN202180009367.8A priority Critical patent/CN114981295A/zh
Publication of WO2021143810A1 publication Critical patent/WO2021143810A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/26—Glucagons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
    • 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

Definitions

  • the present invention relates to the field of medicine. More specifically, the present invention relates to agonizing the glucagon-like peptide-1 (GLP-1) receptor and optionally agonizing the glucose-dependent insulinotropic polypeptide (GIP) receptor and glucagon GCG receptor double or triple agonist polypeptide drug and its application for the treatment of metabolic syndrome (such as diabetes, obesity, non-alcoholic fatty liver).
  • GLP-1 glucagon-like peptide-1
  • GIP glucose-dependent insulinotropic polypeptide
  • Type 2 diabetes and obesity are a type of energy metabolism disorders, which continue to become more and more serious health problems in many countries, and cause a wide range of related dangerous diseases (such as cardiovascular and cerebrovascular diseases, kidney diseases, dyslipidemia, liver diseases, Osteoporosis).
  • Type 2 diabetes is characterized by insulin resistance and hyperglycemia. Obesity is the main cause of insulin resistance, and obesity and insulin resistance are the two major causes of non-alcoholic fatty liver disease (NAFLD/NASH). Therefore, there is an increasing need to find safe and effective drugs to treat these metabolic disorders-related diseases.
  • NAFLD/NASH non-alcoholic fatty liver disease
  • Secretin is a kind of substance secreted from the intestine after eating stimulation under normal physiological conditions. It can rely on the glucose level to stimulate the pancreatic ⁇ -cells to secrete insulin, regulate the homeostasis of glucose levels, and protect the pancreatic ⁇ -cells.
  • GLP-1 and GIP are two types of incretins (Glucagon-like peptide-1 and glucose-dependent insulin-releasing polypeptide plasma levels in response to nutrients. Digestion 1995; 56: 117-126.) that are currently discovered.
  • GLP-1 is expressed in intestinal mucosal L cells by the proglucagon gene. It has a 31-amino acid polypeptide and mainly acts on the GLP-1 receptor (GLP-1 R), which stimulates insulin secretion and inhibits glucagon It secretes and protects pancreatic ⁇ -cells. It has the physiological role of regulating blood sugar homeostasis.
  • Glucagon-like peptide-1 7-36 a physiological incremental in man.Lancet.1987; 2:1300-1304.; Glucagon-like peptide-1 receptor signaling modulates beta cell apoptosis.J Biol Chem.2003;278:471-478.;Relation between gastric empty of glucose and plasma concentrations of glucagon-like peptide-1. Peptides. 1998; 19:1049-1053.).
  • Natural GLP-1 is easily degraded by dipeptidyl peptidase-IV (DPP-IV) and neutral endopeptidase (NEP) that are ubiquitous in plasma, with a half-life of less than 2 min.
  • Exendin-4 (Exendin-4) is a GLP-1 analogue extracted from the salivary glands of African poisonous lizards. It has a stronger GLP-1 receptor agonistic effect and a similar hypoglycemic effect of GLP-1. Compared with natural GLP-1, Exendin-4 has stronger resistance to DPP-4 and NEP, and its half-life and action time in vivo are also longer.
  • Exenatide (trade name ) Is the first GLP-1 drug to be developed and marketed, and it has shown a good clinical effect on diabetes treatment, but it still has greater human immunogenicity and insufficient administration twice a day.
  • amino acid sequence (SEQ ID NO: 2) of GLP-1(7-37) is as follows:
  • GLP-1 analogues In order to improve the half-life of GLP-1 and the effect of diabetes treatment, a variety of long-acting GLP-1 analogues have been successfully developed on the market, such as Novo Nordisk's liraglutide (trade name) for the treatment of diabetes and/or obesity and ) And Somaglutide (trade name ) And Eli Lilly's dulaglutide (trade name ).
  • Liraglutide uses C 16 fatty acids to chemically modify the Lys side chain at position 20 of GLP-1 to increase albumin binding capacity and extend its half-life to 13-16h.
  • the amino acid sequence of liraglutide (SEQ ID NO: 3) is as follows:
  • Somaglutide uses the unnatural amino acid isoaminobutyric acid (Aib) to replace Ala at position 2, which significantly improves DPP-IV resistance. At the same time, the C 18 fatty diacid side chain further enhances the albumin binding force, and the half-life is significantly extended to achieve Dosing once a week.
  • the amino acid sequence of Somaglutide (SEQ ID NO: 4) is as follows:
  • HAibEGTFTSDVSSYLEGQAAK [2-(2-Amino-ethoxy)-ethoxy]-acetyl) 2 - ⁇ Glu-octadecanoic acid acyl group) EFIAWLVRGRG-OH.
  • GIP is a 42 amino acid single-chain polypeptide, produced by small intestinal mucosal K cells, and mainly acts on the GIP receptor (GIP R) in pancreatic islet cells and adipocytes.
  • GIP GIP receptor
  • GIP GIP glucose-dependently promotes insulin secretion, enhances the quality of islet ⁇ -cells, stimulates insulin secretion, inhibits gastric acid secretion, and slows gastric motility.
  • GIP GIP glucose-dependently promotes insulin secretion, enhances the quality of islet ⁇ -cells, stimulates insulin secretion, inhibits gastric acid secretion, and slows gastric motility.
  • it also stimulates the uptake and utilization of fatty acids by adipose tissue cells (Biology of Incretins:
  • GIP also has the physiological effects of promoting osteoblast differentiation, inhibiting osteoblast apoptosis, inhibiting bone resorption, and increasing bone mineral density, and plays a role in protecting bone (Glucose-dependent insulinotropic peptide is an integral hormone with osteotropic effects. Mol Cell Endocrinol.2001;177:35–41.;Effects of glucose-dependent insulinotropic peptide on osteoclast function.AmJ Physiol 2007;292:E543–E548.).
  • amino acid sequence of GIP (SEQ ID NO: 5) is as follows:
  • GCG is a 29-amino acid polypeptide. It is expressed and secreted by the proglucagon gene in pancreatic islet ⁇ -cells. It acts on the glucagon receptor (GCG R) mainly distributed in the liver and kidney to stimulate the decomposition of liver glycogen. , Raise blood sugar, activate lipase, promote fat decomposition, at the same time strengthen fatty acid oxidation, increase the production of ketone bodies. Research results show that GCG has a certain effect on reducing food intake, increasing adipose tissue energy consumption, and reducing body fat (The Metabolic actions of glucagon revised. Nat. Rev. Endocrinol.
  • amino acid sequence of GCG (SEQ ID NO: 6) is as follows:
  • Obesity is a state in which body fat, especially triglycerides, accumulates excessively due to excessive energy intake or abnormal metabolism of the body. Fat accumulation on the pancreas impairs the function of islet cells and aggravates diabetes. Accumulation on the liver can lead to non-alcoholic fatty liver disease.
  • the advantage of the currently marketed GLP-1 analogues is that they have both cardiovascular benefits and weight management effects while lowering blood sugar. However, the clinical maximum weight reduction effects of liraglutide and somaglutide are only 3% and 5%, respectively. Left and right, and gastrointestinal side effects (mainly nausea, vomiting, diarrhea) are more common. Therefore, for more and more obese people, there is still an urgent need for therapeutic drugs with better weight control effects, wider benefits, and greater safety ranges.
  • GLP-1/GCG co-agonists are more effective than a single GLP-1 receptor agonist in reducing food intake, reducing body weight, improving glucose tolerance and reducing triglycerides, and the therapeutic effect on obese mice is more significant (Glucagon- like peptide 1/glucagon receptor dual agonism reverses obesity in mice.Diabetes.2009; 58: 2258-2266; A new glucagon and GLP-1 coagonist eliminates obesity in rodents. Nat Chem Biol. 2009; 5: 749-757.
  • GLP-1 Acyl-GLP-1
  • acetylated GIP Acyl-GIP
  • liraglutide a group consisting of lipid metabolism indicators, such as triglycerides, leptin, adiponectin, ketone bodies, and FGF-21 (Unimolecular Dual Incretins Maximize Metabolic Benefits in Rodents ,Monkeys, and Humans. Sci Transl Med. 2013; 5: 209ra151; A rationally designed monomeric peptide triagonist corrects obesity and diabetes in rodents. Nat Med. 2015; 21: 27-36.).
  • the present invention provides a novel polypeptide compound that can have dual or triple agonistic effects.
  • Such polypeptides may have significant GLP-1 R agonistic activity, and may also have GIP R and/or GCG R agonistic activity. They can be used to prevent or treat metabolic syndrome, such as reducing blood sugar, weight, and fat, and have improved stability. sex.
  • R 1 is selected from H, Ac or pGlu
  • X 3 is selected from His, Gln;
  • X 10 is selected from Tyr, Leu, Lys, Cys or ⁇ ;
  • X 12 is selected from Ile, Arg, Lys, Cys or ⁇ ;
  • X 13 is selected from Tyr, Gln, Lys, Cys or ⁇ ;
  • X 14 is selected from Leu, Lys, Cys or ⁇ ;
  • X 15 is selected from Asp or Glu
  • X 16 is selected from Glu, Lys, Cys or ⁇ ;
  • X 17 is selected from Arg, Ile, Gln, Glu, Lys, Cys or ⁇ ;
  • X 18 is selected from Ala or Arg
  • X 19 is selected from Ala, Val or Gln;
  • X 20 is selected from Gln or Arg
  • X 21 is selected from Asp or Leu;
  • X 23 is selected from Val or Ile
  • X 24 is selected from Glu or Gln
  • X 27 is selected from Leu or Lys
  • X 28 is selected from Ala or Asp
  • R 2 is NH 2 or OH, or a pharmaceutically acceptable salt and/or ester thereof;
  • X 10 , X 12 , X 13 , X 14 , X 16 , and X 17 are ⁇ , and the ⁇ is Cys or Lys whose side chain is modified by the structure having the following formula II, so The formula II mentioned is:
  • Y is selected from the group consisting of Glu (preferably ⁇ Glu), AEEAc, GABA, GSEGSEE, and any combination of two and/or more thereof, and its carboxyl end is connected to the ⁇ -amino group of the side chain of Lys Connected, Z is -CO-(CH 2 ) m -R 3 , m is an integer between 6-24, R 3 is selected from -CH 3 or -COOH; Y preferably represents at most 10, or at most 5, or Up to 4, or up to 3, or up to 2, or 1 linker selected from Glu (preferably ⁇ Glu), AEEAc, GABA, GSEGSEE, and any combination of two and/or more thereof; for example, Y It can be ⁇ Glu, GSEGSEE, AEEAc-AEEAc- ⁇ Glu, ⁇ Glu- ⁇ Glu-AEEAc-AEEAc, ⁇ Glu- ⁇ Glu-AEEAc-AEEAc- ⁇ Glu, or ⁇
  • ⁇ is Cys
  • Y is Y1-Y2
  • Y1 is selected from acetylglycyl, 3-maleimidopropionyl and any combination thereof
  • Y2 is selected from Glu (preferably ⁇ Glu), AEEAc and Any combination thereof, and it is connected to the Cys side chain sulfhydryl group through acetylglycyl or 3-maleimidopropionyl
  • Z is -NH-(CH 2 ) m -R 3
  • m is between 6-24
  • R 3 is selected from -CH 3 or -COOH
  • Y1 preferably represents a linker selected from acetylglycyl, 3-maleimidopropionyl and any combination thereof
  • Y2 preferably represents the most 10, or at most 5, or at most 4, or at most 3, or at most 2, or 1 linker selected from Glu (preferably ⁇ Glu), AEEAc and any combination thereof; for example, Y1 can be 3- Maleimi
  • amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% similarity or identity to the amino acid sequence.
  • R 1 is selected from H, Ac or pGlu
  • X 3 is selected from His, Gln;
  • X 10 is selected from Leu, Lys, Cys or ⁇ ;
  • X 13 is selected from Tyr or Gln;
  • X 14 is selected from Leu, Lys, Cys or ⁇ ;
  • X 15 is selected from Asp or Glu
  • X 17 is selected from Arg, Gln or Glu
  • X 19 is selected from Ala or Val
  • X 20 is selected from Gln or Arg
  • X 21 is selected from Asp or Leu
  • X 23 is selected from Val or Ile
  • X 24 is selected from Glu or Gln
  • X 27 is selected from Leu or Lys
  • X 28 is selected from Ala or Asp
  • R 2 is NH 2 or OH, or a pharmaceutically acceptable salt and/or ester thereof;
  • X 10 and X 14 are ⁇ , and the ⁇ is Lys whose side chain is modified by the structure having the following formula II, and the formula II is:
  • Y is selected from Glu (preferably ⁇ Glu), AEEAc, GABA, GSEGSEE and any combination of two and/or more thereof, and its carboxyl end is connected to the ⁇ -amino group of the side chain of Lys, Z is -CO-(CH 2 ) m -R 3 , m is an integer between 6-24, R 3 is selected from -CH 3 or -COOH; Y preferably represents at most 10, or at most 5, or at most 4, or at most 3, Or at most 2, or 1 linker selected from Glu (preferably ⁇ Glu), AEEAc, GABA, GSEGSEE, and any combination of two and/or more thereof; for example, Y can be ⁇ Glu, GSEGSEE, AEEAc-AEEAc - ⁇ Glu, ⁇ Glu- ⁇ Glu-AEEAc-AEEAc, ⁇ Glu- ⁇ Glu-AEEAc-AEEAc- ⁇ Glu, or ⁇ Glu-GABA-AEEAc- ⁇ Glu
  • amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% similarity or identity to the amino acid sequence.
  • the above compounds with the structure of formula Ib can be prepared by chemical synthesis of polypeptides.
  • R 1 His-Aib-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Tyr-X 14 -Glu-Glu-Gln-Ala-Ala-Gln-Asp-Phe-Ile- Glu-Trp-Leu-Leu-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-R 2 (Ic) (SEQ ID NO:47)
  • R 1 is H
  • X 14 is ⁇
  • R 2 is NH 2 or OH, or a pharmaceutically acceptable salt and/or ester thereof;
  • X 14 is ⁇
  • ⁇ is Cys whose side chain is modified by the structure having the following formula II, and the formula II is:
  • Y is Y1-Y2, Y1 is selected from acetylglycyl, 3-maleimidopropionyl and any combination thereof, Y2 is selected from Glu (preferably ⁇ Glu), AEEAc and any combination thereof, and it passes through the acetyl group Glycyl or 3-maleimidopropionyl is connected to Cys side chain sulfhydryl group, Z is -NH-(CH 2 ) m -R 3 , m is an integer between 6-24, R 3 is selected from- CH 3 or -COOH; Y1 preferably represents a linker selected from acetylglycyl, 3-maleimidopropionyl and any combination thereof; Y2 preferably represents at most 10, or at most 5, or Up to 4, or up to 3, or up to 2, or 1 linker selected from Glu (preferably ⁇ Glu), AEEAc and any combination thereof; for example, Y1 can be 3-maleimidopropionyl; Y2 can
  • amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% similarity or identity to the amino acid sequence.
  • R 1 is H
  • X 3 is selected from His, Gln;
  • X 10 is selected from Tyr or Leu;
  • X 12 is selected from Ile, Arg, or ⁇ ;
  • X 13 is selected from Tyr, Gln, or ⁇ ;
  • X 14 is selected from Leu, or ⁇ ;
  • X 16 is selected from Glu, or ⁇ ;
  • X 17 is selected from Arg, Ile, Gln, or ⁇ ;
  • X 18 is selected from Ala or Arg
  • X 19 is selected from Ala or Gln;
  • X 23 is selected from Val or Ile
  • X 28 is selected from Ala or Asp
  • R 2 is NH 2 or OH, or a pharmaceutically acceptable salt and/or ester thereof;
  • X 12 , X 13 , X 14 , X 16 , X 17 and only one is ⁇
  • the ⁇ is Lys whose side chain is modified by the structure having the following formula II, and the formula II is :
  • Y is selected from Glu (preferably ⁇ Glu), AEEAc, GABA, GSEGSEE and any combination of two and/or more thereof, and its carboxyl end is connected to the ⁇ -amino group of the side chain of Lys, Z is -CO-(CH 2 ) m -R 3 , m is an integer between 6-24, R 3 is selected from -CH 3 or -COOH; Y preferably represents at most 10, or at most 5, or at most 4, or at most 3, Or at most 2, or 1 linker selected from Glu (preferably ⁇ Glu), AEEAc, GABA, GSEGSEE, and any combination of two and/or more thereof; for example, Y can be ⁇ Glu, GSEGSEE, AEEAc-AEEAc - ⁇ Glu, ⁇ Glu- ⁇ Glu-AEEAc-AEEAc, ⁇ Glu- ⁇ Glu-AEEAc-AEEAc- ⁇ Glu, or ⁇ Glu-GABA-AEEAc- ⁇ Glu;
  • amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% similarity or identity to the amino acid sequence.
  • the above-mentioned compounds with the structure of Formula Id can be prepared by chemical synthesis of polypeptides or biological semi-synthesis.
  • the biological semi-synthesis refers to a method similar to that disclosed in patents CN201510459093 and US9732137 for producing somaglutide (SEQ ID NO: 4).
  • the method features: firstly use yeast or E. coli to ferment and express part of the peptide TFTSDVSSYLEGQAAKEFIAWLVRGRG-OH in the somaglutide sequence; then use fatty acid activated ester to react with the side chain NH 2 of K in the peptide to obtain the peptide.
  • TFTSDVSSYLEGQAAK ([2-(2-Amino-ethoxy)-ethoxy]-acetyl)2- ⁇ E-octadecanoic acid acyl)EFIAWLVRGRG-OH; finally, use the N-terminal overhang Boc-His(Boc )-Aib-Glu(O-tBu)-Gly-OSuc reacts with the ⁇ -NH 2 acylation reaction of the above-mentioned peptide, and removes the Boc protecting group to obtain somaglutide.
  • This method overcomes the cumbersome steps of chemical synthesis, makes full use of the advantages of biological fermentation expression, and is beneficial to the reduction of production costs and large-scale industrial production.
  • R 1 is H
  • X 3 is selected from His, Gln;
  • X 10 is selected from Tyr or Leu;
  • X 12 is selected from Ile, Arg, Lys, Cys or ⁇ ;
  • X 13 is selected from Tyr, Gln, Lys, Cys or ⁇ ;
  • X 14 is selected from Leu, Lys, Cys or ⁇ ;
  • X 16 is selected from Glu, Lys, Cys or ⁇ ;
  • X 17 is selected from Arg, Ile, Gln, Lys, Cys or ⁇ ;
  • X 18 is selected from Ala or Arg
  • X 19 is selected from Ala or Gln;
  • X 23 is selected from Val or Ile
  • X 28 is selected from Ala or Asp
  • R 2 is NH 2 or OH, or a pharmaceutically acceptable salt and/or ester thereof;
  • X 12 , X 13 , X 14 , X 16 , X 17 and only one is ⁇
  • the ⁇ is Lys whose side chain is modified by the structure having the following formula II, and the formula II is :
  • Y is selected from Glu (preferably ⁇ Glu), AEEAc, GABA, GSEGSEE and any combination of two and/or more thereof, and its carboxyl end is connected to the ⁇ -amino group of the side chain of Lys, Z is -CO-(CH 2 ) m -R 3 , m is an integer between 6-24, R 3 is selected from -CH 3 or -COOH; Y preferably represents at most 10, or at most 5, or at most 4, or at most 3, Or at most 2, or 1 linker selected from Glu (preferably ⁇ Glu), AEEAc, GABA, GSEGSEE, and any combination of two and/or more thereof; for example, Y can be ⁇ Glu, GSEGSEE, AEEAc-AEEAc - ⁇ Glu, ⁇ Glu- ⁇ Glu-AEEAc-AEEAc, ⁇ Glu- ⁇ Glu-AEEAc-AEEAc- ⁇ Glu, or ⁇ Glu-GABA-AEEAc- ⁇ Glu;
  • amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% similarity or identity to the amino acid sequence.
  • the above formula II has the following structure:
  • the structure of formula II can be selected from (the term "R" in the following structure is intended to indicate the connection site of the peptide backbone to the formula II, that is ⁇ -amino group of Lys side chain):
  • the structure of formula II can be selected from (the term "R" in the following structures is intended to indicate the connection site of the peptide backbone to the formula II, that is Cys side chain mercapto group):
  • the compound is selected from:
  • a pharmaceutical composition which includes an effective amount of the compound or a salt or solvate of any one of the foregoing aspects, and pharmaceutically acceptable excipients, diluents, carriers or excipients.
  • the pharmaceutical composition is injection or lyophilized powder, tablet, pill, lozenge, soft capsule, hard capsule, granule, powder, solution, suspension or syrup; or
  • the pharmaceutical composition is in the form of microcapsules, microspheres, nanoparticles or liposomes.
  • the pharmaceutical composition is for oral administration, inhalation administration or parenteral administration, and the parenteral administration is selected from the group consisting of intraperitoneal, intramuscular, intraarterial, intravenous, subcutaneous or intradermal injection. Medicine.
  • the pharmaceutical composition is administered at least once a day, once a week, or once a month.
  • the metabolic syndrome is selected from high Glycemia, insulin resistance, impaired glucose tolerance, type 2 diabetes, obesity or non-alcoholic fatty liver disease/non-alcoholic steatohepatitis (NAFLD/NASH), diabetic nephropathy, diabetic retinopathy, dyslipidemia, osteoporosis; Or a neurodegenerative disease selected from Alzheimer's disease or Parkinson's syndrome.
  • a method for agonizing one, two or three of GLP-1 receptor, GIP receptor or GCG receptor comprises administering to an individual in need an effective amount of the compound of the present invention or its Salt or solvate.
  • metabolic syndrome being selected from hyperglycemia, insulin resistance, glucose intolerance, type 2 diabetes, obesity or Non-alcoholic fatty liver disease/non-alcoholic steatohepatitis (NAFLD/NASH), diabetic nephropathy, diabetic retinopathy, dyslipidemia, osteoporosis; or neurodegenerative disease selected from Alzheimer’s disease Or Parkinson's syndrome, including administering an effective amount of the compound of the present
  • the compound of the present invention can be a double- or tri-agonistic polypeptide molecule, which can be used to prevent or treat abnormal metabolic syndrome and has improved stability.
  • Figure 2 The electropherogram of the precursor fusion protein of KSI-DDDDK-peptide SEQ ID NO.19 (5-40).
  • FIG. 5 Effects of compounds on oral glucose tolerance in DIO mice.
  • the glucose tolerance of DIO mice was measured.
  • the blood glucose content at a specific time point within 120 minutes was measured, and the area under the blood glucose-time curve (AUC 0-120min ) was calculated.
  • Figure 5A Blood glucose-time curve after oral glucose
  • Figure 5B Blood glucose-time curve after oral glucose (AUC 0-120min ).
  • the compound significantly reduced the glucose tolerance of DIO mice after oral administration of glucose (***p ⁇ 0.001). Data are expressed as mean ⁇ SEM, n 8.
  • the amino acid sequence of the parent peptide of the polypeptide drug of the present invention may be based on Exendin-4 (SEQ ID NO: 1), GLP-1 (SEQ ID NO: 2), GIP (SEQ ID NO: 5), GCG ( The amino acid sequence of SEQ ID NO: 6) is modified.
  • polypeptide compounds of the present invention can be synthesized and modified by those skilled in the art through known technical methods.
  • the peptide sequence backbone of the polypeptide compound of the present invention can be prepared by synthetic methods, recombinant DNA biotechnology and other methods.
  • the peptide backbone of the compound of the present invention is chemically modified by a fatty acid side chain group at at least one site.
  • the compound may have a stable peptide ⁇ -helical structure, and may have an enhanced albumin binding capacity, which can achieve improved stability of the peptide compound and prolonged peptide action time.
  • the compound of the present invention has agonistic activity on GLP-1 receptor, and optionally also has agonistic activity on GIP receptor and/or GCG receptor.
  • the "agonistic activity” means that the compound can stimulate specific receptor cells to produce cAMP, and the cells used can be host cells or pancreatic islets that overexpress GLP-1 receptor or GIP receptor or GCG receptor constructed by those skilled in the art. Tissue cells, fat cells, liver cells, etc.
  • the receptor agonistic activity can be measured by the EC 50 value of the compound agonizing the receptor cells to produce cAMP.
  • the EC 50 value is the concentration of the drug required to reach half of the maximum activity (50% activity) of the compound in a specific assay system.
  • the agonistic activity of a compound can be evaluated by evaluating the relative activity of a specific natural compound. 50 percent relative activity of the particular value of EC EC 50 value of natural compounds in the ratio of the test compound.
  • the relative activity of the compound of the present invention on GLP-1 receptor agonism is at least 0.5%, preferably at least 5%, more preferably at least 50%, even most preferably at least 100% .
  • the relative activity of the compound of the present invention on GIP receptor agonism is at least 0.5%, preferably at least 5%, more preferably at least 50%, and even most preferably at least 100%.
  • the relative activity of the compound of the present invention on GCG receptor agonism is at least 0.5%, preferably at least 5%, more preferably at least 50%, and even most preferably at least 100%.
  • the structural similarity of two polypeptides can be determined by aligning the residues of two polypeptides (such as the candidate polypeptides described herein and any suitable reference polypeptides) along their sequence lengths to optimize the number of identical amino acids; In order to optimize the number of identical amino acids in the alignment, gaps in any one or two sequences are allowed, but the amino acids in each sequence must still retain its correct order.
  • the reference polypeptide may be the polypeptide described herein.
  • the candidate polypeptide is a polypeptide that is compared with a reference polypeptide.
  • non-polar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and tyrosine.
  • Polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine.
  • the positively charged (basic) amino acids include arginine, lysine and histidine.
  • Negatively charged (acidic) amino acids include aspartic acid and glutamic acid.
  • Conservative substitutions include, for example, substitution of Lys for Arg and vice versa to maintain a positive charge; Glu for Asp and vice versa to maintain a negative charge; Ser for Thr to maintain free -OH; and Gln for Asn to maintain free -NH 2 .
  • biologically active analogs of polypeptides are also considered, which contain the deletion or addition of one or more adjacent or non-adjacent amino acids that do not eliminate the functional activity of the polypeptide.
  • the amino acid sequence referring to the polypeptide of the present invention and/or referring to one or more SEQ ID NOs may include polypeptides having the following amino acid sequence similarity with the reference amino acid sequence: at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92% , At least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%.
  • the amino acid sequence referring to the polypeptide of the present invention and/or one or more SEQ ID NOs may include polypeptides having the following amino acid sequence identity with the reference amino acid sequence: at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92% , At least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%.
  • the polypeptide of the present invention may be further modified, for example, including a chemically or enzymatically derived polypeptide (or an analog thereof, such as a fragment thereof) at one or more constituent amino acids.
  • the modification may include, for example, side chain modification, main chain modification, and N- and C-terminal modification, such as acetylation, hydroxylation, methylation, amidation, and attachment of carbohydrate or lipid moieties, cofactors, etc., and their The combination.
  • the modified polypeptide of the present invention may retain the biological activity of the unmodified polypeptide, or may exhibit reduced or increased biological activity.
  • the effective amount of the compound of the present invention can be combined with at least one pharmaceutically acceptable auxiliary material, diluent, carrier or excipient to form a pharmaceutical composition.
  • Effective amount of compound refers to the amount of compound that can produce disease alleviation or treatment, but not produce damaging effects.
  • the effective amount can be appropriately determined by the attending physician according to the patient's disease severity, age, sex, weight, general health status, and the like.
  • the patient is a mammal.
  • the mammal is selected from cows, horses, goats, sheep, dogs, chimpanzees, rabbits, mice, rats, monkeys, pigs, and humans.
  • the patient is a human.
  • the compound of the present invention and its pharmaceutical composition can be made into injections or freeze-dried powders, tablets, pills, lozenges, soft capsules, hard capsules, granules, powders, solutions, suspensions or syrups, Preferably, injection or lyophilized powder.
  • Excipients that can be used in the composition of the present invention such as lubricants, binders, fillers, preservatives, surfactants, colorants, flavoring agents, emulsifiers, suspending agents, diluents, gelling agents, disintegrants Agents, pH adjusters, solubilizers, etc.
  • lubricants such as lubricants, binders, fillers, preservatives, surfactants, colorants, flavoring agents, emulsifiers, suspending agents, diluents, gelling agents, disintegrants Agents, pH adjusters, solubilizers, etc.
  • the compound of the present invention can also be loaded into a variety of drug carrier materials (for example, microcapsules, microspheres, nanoparticles, liposomes) or drug delivery devices for use.
  • drug carrier materials for example, microcapsules, microspheres, nanoparticles, liposomes
  • drug delivery devices for use.
  • the compound of the present invention can also be used in combination with at least one of the following therapeutic active agents, including anti-diabetic active agents (such as: insulin and its analogs, biguanides, sulfonylureas, thiazolidinediones, ⁇ -Glucosidase inhibitor, DPP-4 inhibitor, SGLT2 inhibitor, dual SGLT1/SGLT2 inhibitor, GLP-1 receptor agonist, amylin and its analogs), GIP receptor agonist, GCG receptor Agonist or antagonist, dual GLP-1/GIP receptor agonist, GLP-1/GCG receptor agonist, GIP/GCG receptor agonist, FGF-21 and its analogs, cholecystokinin B (CCKB) And its analogues, PYY(3-36) and its analogues, leptin and its analogues, calcitonin and its analogues, active drugs for regulating blood lipids, PPAR- ⁇ , ⁇ , ⁇
  • the compound of the present invention can promote insulin secretion and lower blood sugar.
  • food intake can be suppressed, gastric emptying can be delayed, energy consumption can be increased, and the effect of weight reduction can be observed in the end.
  • the compound of the present invention can reduce pancreatic ⁇ -cell apoptosis, increase the number of pancreatic ⁇ -cells, and improve the function of pancreatic islet cells.
  • the compound of the present invention can also improve blood lipids, reduce liver fat accumulation, inhibit the development of liver inflammation, and prevent and treat non-alcoholic fatty liver disease.
  • the compound of the present invention is expected to promote the growth of brain neurons, eliminate neurotoxic substances, inhibit the development of inflammation, and play a neuroprotective effect.
  • the compound or composition of the present invention can be used to prevent and/or treat metabolic disorders and related complications. It is preferably used for the treatment of diabetes, obesity and non-alcoholic fatty liver disease.
  • the compound or composition of the present invention can be used to treat dyslipidemia and related diseases, neurodegenerative diseases (e.g., Parkinson's disease, Alzheimer's disease).
  • neurodegenerative diseases e.g., Parkinson's disease, Alzheimer's disease.
  • the compound or composition of the present invention can be used to treat endocrine diseases, metabolic disorders, kidney diseases, weight loss and other causes-related skeletal diseases, such as osteoporosis, osteoarthritis.
  • the compound or composition of the present invention can be administered by various routes, for example, it can be used for oral administration, inhalation administration or parenteral administration, such as intraperitoneal, intramuscular, and intraarterial administration. , Intravenous, subcutaneous or intradermal injection.
  • the compound or composition of the present invention can be administered at least once a day, once a week, or once a month.
  • the compound of the present invention exhibits better solubility and stability, which is better than natural peptide molecules and liraglutide.
  • the compound of the present invention has a significant agonistic effect on two or three of GLP-1, GIP, and GCG receptors.
  • GABA ⁇ -aminobutyric acid
  • AEEAc [2-(2-Amino-ethoxy)-ethoxy]-acetyl
  • PEG polyethylene glycol
  • Trt Trityl
  • Tris Tris
  • DIPEA N,N-Diisopropylethylamine
  • PAM Peptidylglycine ⁇ -amidating monooxygenase
  • HEK-293 Human embryonic kidney cells
  • GLP-1 R Glucagon-like peptide-1 receptor
  • GIPR Glucose-dependent insulin releasing peptide receptor
  • FBS Fetal Bovine Serum
  • DMEM Dulbecco's Modified Eagle Medium
  • BSA Bovine Serum Albumin
  • IBMX 3-isobutyl-1-methylxanthine.
  • LDL-C Low-density lipoprotein cholesterol
  • HOMA Insulin Resistance Index
  • the intermediates and compounds of the present invention can be synthesized and prepared by various methods known in the art.
  • the following specific examples illustrate the preparation of the compounds of the present invention by chemical synthesis methods. Each specific synthesis step described can be combined with different materials and methods to synthesize multiple corresponding compounds of the present invention or their salts.
  • the reagents and raw materials used are easily available to those of ordinary skill in the art.
  • the following examples are only used to illustrate the present invention, and should not limit the scope of the present invention in any way.
  • the materials and reagents used in the present invention are all purchased from commercial products, and the protective amino acids used in the entire synthesis process are as follows: Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Ala-OH, Fmoc-Gly-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Leu-OH, Fmoc-Trp(Boc)-OH, Fmoc-Gln(Trt)-OH, Fmoc-Val-OH, Fmoc-Phe-OH, Fmoc-Arg(pbf )-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Tyr(t-Bu)-OH, Fmoc-Cys(Trt)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Lys(ivdde)-OH , Fmoc
  • the compound of SEQ ID NO: 12 is taken as an example to illustrate the synthetic preparation method of the compound of the present invention.
  • the Lys at the side chain modification site is replaced with Fmoc-Lys(ivdde)-OH.
  • Lys side chain modification add the prepared Fmoc-Glu-OtBu solution to the above-mentioned processed resin, add DIC/DMF solution, and stir for 1 hour. After the reaction is completed, filter and wash, add 20% piperidine/DMF solution, stir and react for 20 minutes to remove the Fmoc group. Then add the prepared palmitic acid solution to the resin, add the DIC/DMF solution, and stir and react for 1 hour. After the reaction is completed, the resin is washed 4 times, and the resin is drained.
  • Post-treatment of peptide resin Add cutting reagent K to cut the resin. After filtering the filtrate, the filtrate was precipitated and centrifuged to obtain a white solid as the crude product of the target compound.
  • the obtained crude peptide was purified by a reverse-phase C18 preparative chromatographic column (Shimadzu, Inertsil ODS 20x 250mm 5um), and the sample was purified to a purity greater than 95%.
  • 95% buffer A 0.065% TFA/H 2 O
  • 5% buffer B 0.05% TFA/acetonitrile
  • the purified peptide compound was analyzed and confirmed by analytical HPLC/MS method.
  • the following takes the biological semisynthesis of the peptide compound SEQ ID NO. 19 as an example to illustrate the method steps of the biological semisynthesis of the compound of the present invention.
  • the fusion gene has a gene sequence shaped like an ABC structure, where A is the chaperone protein coding gene, B is the linker peptide coding gene, and C is the tri-agonist peptide SEQ ID NO.19 (5-40) fragment encoding gene. It is subjected to high-density fermentation, induced, and the inclusion body or fusion protein containing the fusion protein is extracted.
  • the recombinant E. coli strain is preferably obtained by the following steps: cloning the fusion gene in the shape of A-B-C structure into a prokaryotic expression vector, and then transferring the obtained recombinant expression vector into an engineered E. coli strain to obtain a recombinant E. coli strain.
  • the prokaryotic expression vector is pET31b(+).
  • the engineered Escherichia coli bacteria is Escherichia coli BL21 (DE3).
  • the induction is induced by IPTG.
  • the fusion protein has the following structure, that is, from the N-terminus to the C-terminus, three fragments of the KSI chaperone protein, the connecting peptide and the peptide SEQ ID NO.19 (5-40) precursor are connected.
  • amino acid sequence of the fusion protein is shown in SEQ ID NO. 41.
  • the connecting peptide in the fusion protein is DDDDK.
  • the peptide SEQ ID NO. 19 (5-40) precursor sequence of the fusion protein is shown in SEQ ID NO. 42.
  • the construction method of the fusion gene and engineered bacteria with the A-B-C structure can refer to the experimental guide in this field (J. Sambrook et al., "Molecular Cloning Experiment Guide” Second Edition, Science Press, 1995).
  • Design DDDDK-peptide SEQ ID NO.19(5-40) precursor fusion gene fragment convert these amino acid sequences into nucleotide sequences according to the codon table, and select according to the preference of E. coli codon usage during the conversion process Use more frequent codons, adjust their GC content, remove cis-acting elements and repetitive sequences that affect gene transcription, and optimize them. At the same time, introduce the double stop codon TAATGA at the 3'end of the gene sequence.
  • DDDDK-peptide SEQ ID NO.19 (5-40) precursor fusion gene sequence is introduced into the 5'end of the AlwNI restriction site sequence CAGATGCTG, so two amino acids of ML are introduced before the N-terminal extension peptide, in the DDDDK-peptide SEQ ID
  • the 3'end of NO.19(5-40) precursor fusion gene introduces the XhoI restriction site CTCGAG, and the optimized DDDDK-peptide SEQ ID NO. 19(5-40) precursor fusion gene sequence is as SEQ ID NO: 43 shown.
  • the gene sequence was synthesized by a gene synthesis service company, and TA cloned into pUC57 vector.
  • Competent cells of Escherichia coli BL21 (DE3) (both purchased from Life Technologies) were prepared according to the calcium chloride method provided in the third edition of the "Molecular Cloning Experiment Guide” published by Cold Spring Harbor Laboratory in the United States. 1 ⁇ L of the recombinant expression vector pET31b-SEQ ID NO.19 was transformed into E. coli BL21(DE3) competent cells, and the transformation method was also carried out in accordance with the calcium chloride method in the third edition of the "Molecular Cloning Experiment Guide”.
  • Example 6 of patent CN201711154044 the constructed engineering bacteria are subjected to high-density fermentation, and the bacteria are broken and washed to obtain inclusion bodies.
  • the 10L fermentation broth finally obtained 0.62kg of inclusion bodies, and the Folin-phenol method showed that the amount of protein per 1g of inclusion bodies was 0.20g.
  • Dissolve 200g of the above fusion protein inclusion body in 1500ml 6mol/L guanidine hydrochloride solution add 80ml DMSO and 15ml N,N-diisopropylethylamine, mix well, pH 10.8; then add 50mg/ml N ⁇ -ten 80ml of hexaacyl-Glu(ONSu)-OH in DMSO solution, stirred for 3 hours; then add 1700ml 20mmol/L tris solution, and add 900IU lysyl specific endonuclease, dilute hydrochloric acid or Sodium hydroxide solution was used to adjust the pH to 9.0, and after 8 hours of reaction, the pH was adjusted to 7.8.
  • the collected components were diluted with one-fold volume of purified water, and the sample was loaded to 500ml UniPS40 (particle size 40 ⁇ m, 500ml UniPS40 (particle size 40 ⁇ m, 1500ml) containing 20mmol/LTris, 5% isopropanol, and pH 7.0 balance solvent.
  • 500 angstroms (pore size) in the chromatography column and then equilibrate the chromatography column with 1500 ml of equilibrium solvent. Elution was performed with a gradient elution solvent containing 20 mmol/LTris, 5% to 30% isopropanol pH 7.0, and the fractions containing peptide SEQ ID NO. 19 (5-40) were collected. And lyophilize it into powder.
  • SEQ ID NO. 19 The sequence of the peptide SEQ ID NO. 19 (5-40) is shown in SEQ ID NO. 44.
  • Example 4 of patent CN201510459093 and respectively couple Fmoc-Gly-OH, Fmoc-His(Trt)-OH, Fmoc-Aib-OH, Boc-His(Boc) on 2-chlorotrityl chloride resin in sequence. -OH, finally add trifluoroethanol-DCM (1:4), stir the resin. The filtrate was collected, the solvent was removed in vacuo, cold ether was added, the precipitate was filtered, washed with ether and dried in vacuo to obtain Boc-His(Boc)-Aib-His(Trt)-Gly-OH.
  • Example 5 of patent CN201510459093 to the anhydrous THF solution of Boc-His(Boc)-Aib-His(Trt)-Gly-OH, add a certain amount of DIPEA and hexafluorophosphate O-(N-succinyl Imino)-N,N,N,N-tetramethyluronium.
  • the reaction solution is added with dichloromethane, washed with water, and the organic phase is dried over magnesium sulfate and dried in vacuum to obtain Boc-His(Boc)-Aib-His(Trt)-Gly-Osu.
  • Example 8 of patent CN201510459093 the peptide SEQ ID NO.19(5-40) (0.24mmol, 1.0g) was dissolved in 100ml of NMP, and then 300ul DIPEA and 0.4mmol Boc-His(Boc)-Aib-His were added (Trt)-Gly-Osu, stir the reaction overnight. 1000ml of ice-cold ether was added, the precipitate was separated by centrifugation, washed with 500ml of ether, and then dried in vacuo.
  • the above-mentioned crude dry powder was dissolved in TFA-triisopropylsilane-water (95:2.5:2.5, 200ml) and reacted with stirring for 2 hours, and then the solution was concentrated in vacuo to about 20ml. Then add 500ml of ice-cold ether, stand at 2-8°C for 6-8h to form a precipitate, and centrifuge to precipitate. The precipitate was washed with cold ether and dried under vacuum. The collected dry powder is the crude powder of the peptide SEQ ID NO. 19 (1-40).
  • the purification method is as follows:
  • Phase A 95% 0.1M phosphate buffer + 5% acetonitrile, pH 6.5;
  • HPLC/MS detection on the collected liquid containing peptide sample components.
  • the HPLC method is as follows:
  • Mobile phase A water containing 0.065% TFA
  • mobile phase B acetonitrile containing 0.05% TFA
  • Peptidylglycine ⁇ -amidating monooxygenase (purchased from Wuhan Yunclone Technology Co., Ltd.) was used to catalytically cleave the C-terminal PPPSG-OH structure of the peptide SEQ ID NO.19 (1-40) sequence.
  • the C-terminal structure is the amidation product of PPPS-NH 2.
  • the reaction solution is diluted with 5% acetonitrile-containing pH 7.0 phosphate buffer by one volume.
  • the obtained sample is purified to a purity greater than 95%.
  • the purified samples were characterized and confirmed by analytical HPLC/MS methods, and the peptide compound with the sequence structure SEQ ID NO.19 was obtained.
  • SEQ ID NO. 15-40 peptide compounds can all be semi-synthesized by the above method, wherein the peptide sequence molecules with C-terminal amidation can be prepared by amidation after the semi-synthesis.
  • test compound was dissolved in a newly prepared PBS solution at 1 mg/ml, adjusted to pH 7.4, and filtered with a 0.22 ⁇ m sterile filter. Aspirate the compound solution and analyze the peak area of 10 ⁇ L injection by HPLC-UV. The result of this analysis is the initial point (T 0 ) of the stability test of the compound.
  • the compound sample solution for stability test was placed in a 25° C. thermostat, sealed and protected from light for 7 days. After the process, the sample solution was centrifuged at 4500 rpm for 10 min, the supernatant solution was gently aspirated, and the peak area of the 10 ⁇ L injection was analyzed by HPLC-UV. This analysis result was the end point (T 7 ) of the stability test of the compound.
  • the remaining peptide amount of the measured peptide is calculated. Calculated as follows:
  • Remaining peptide (%) (T 7 main peak area/T 0 main peak area) ⁇ 100
  • the stability of the peptide compound was evaluated by comparing the remaining amount of peptide of the peptide compound, and the measurement results are shown in Table 2.
  • the agonistic activity of the peptide compounds on the corresponding receptors was determined.
  • the intracellular cAMP content was determined using Cisbio Corp.'s kit based on HTRF (homogeneous time-resolved fluorescence) technology.
  • HEK-293 cells stably overexpressing human GLP-1, GIP, and GCG receptors in DMEM complete medium containing 10% FBS and 2mM L-glutamine.
  • DMEM complete medium containing 10% FBS and 2mM L-glutamine.
  • Digest with 0.025% trypsin terminate the digestion with complete medium and gently blow the cell mass into individual cells. Centrifuge the cell liquid at 1000 rpm for 5 min at room temperature, discard the supernatant, and use 1 ⁇ HBSS (20mM HEPES, 0.1% BSA, 250 ⁇ M IBMX). ) Resuspend the cells at a cell density of 1.0 ⁇ 10 5 /mL.
  • GLP-1 receptor cells Use natural wild-type human GLP-1, GIP, GCG as a positive control for the receptor agonistic effect of the test compound.
  • GLP-1 receptor cells calculate the ratio of the EC 50 value of the test compound to the EC 50 value of human GLP-1 The percentage is used as the relative activity (%) to evaluate the GLP-1 receptor agonistic activity of the test compound.
  • the percentage value 50 and the human GIP EC 50 values as a ratio of the relative activity (%) Evaluation test compound GIP receptor agonistic activity of the test compound is calculated by EC.
  • the test compound is calculated by EC as relative activity (%) to assess the percentage of 50 and EC 50 values of the ratio of the value of the GCG GCG human receptor agonistic activity of the test compound.
  • Rel.A Relative activity
  • NT not test
  • n ⁇ 4 Each group has at least 4 independent test data.
  • SD rats received the compound SEQ ID NO: 19 (30, 100 nmol/kg), SEQ ID NO: 39 (50 nmol/kg), SEQ ID NO: 40 (50 nmol/kg), liraglutide (30 nmol/kg), Marglutide (50nmol/kg) was administered by subcutaneous injection.
  • mice in SEQ ID NO: 19 and liraglutide group were administered at 0.25, 0.5, 1, 2, 4, 8, 12, 24, 36, 48, 56h Blood was collected through the jugular vein at the time point, SEQ ID NO: 39, SEQ ID NO: 40, Semaglutide group animals at 0.25, 0.5, 1, 2, 4, 8, 12, 24, 48, 72, 96h time points
  • the blood was collected through the jugular vein, the blood was processed to obtain the plasma sample, and the sample was analyzed by LC-MS/MS.
  • the blood drug concentration-time curve was analyzed using Phoenix WinNonlin version 6.3 software (non-compartmental model), and the PK parameters and half-life were calculated.
  • Example 6 Study on the efficacy of the compound of SEQ ID NO: 36, SEQ ID NO. 37, SEQ ID NO: 38 on diet-induced obesity (DIO) mice
  • mice with high-fat diet induced obesity have obvious metabolic syndrome characteristics similar to humans, such as obesity, elevated blood sugar, insulin resistance, and dyslipidemia.
  • DIO high-fat diet induced obesity
  • the animal is given a vehicle control, a certain dose of semaglutide or the compound of the present invention by subcutaneous injection according to the experimental group.
  • the compound is dissolved in 1 ⁇ PBS, and the dosage is 5ml/kg.
  • the administration operation is at 9 in the morning: Starting at 00, Vehicle, SEQ ID NO: 36 and SEQ ID NO: 37 are administered once a day (QD), and Semaglutide and SEQ ID NO: 38 are administered once every three days (Q3D) for 22 days.
  • QD Vehicle
  • Semaglutide and SEQ ID NO: 38 are administered once every three days (Q3D) for 22 days.
  • Mouse HOMA-IR [(fasting insulin(mIU/L) ⁇ fastingglucose(mmol/l)]/22.5. The result is expressed as the mean ⁇ SEM of 8 animals.
  • Example 7 Pharmacodynamic study of the compounds of SEQ ID NO: 19, SEQ ID NO: 39, SEQ ID NO: 40 on diet-induced obesity (DIO) mice
  • the animal vehicle control Vehicle (1 ⁇ PBS) and a certain dose of the compound of the present invention were given by subcutaneous injection, SEQ ID NO: 19, SEQ ID NO: 39, SEQ ID NO: 40, and the compound was dissolved in 1 ⁇ PBS ,
  • the dosage is 5ml/kg, the dosing operation starts at 9:00 in the morning, the compound SEQ ID NO: 19 is administered once a day (QD), the compound SEQ ID NO: 39, SEQ ID NO: 40 every three days Dosing once (Q3D) for 14 days.
  • the weight and food intake of the animals were measured before administration every day. By comparing with the initial body weight and food intake of the same animal before administration, the percentage of animal weight change (%) and the cumulative food intake were calculated to evaluate the compound's effect on body weight and food intake. The effect of changes in food intake.
  • the animal feed of each group was withdrawn, and the animals were fasted overnight for 12h (cannot help water).
  • Day 15 the animals were weighed at 9:00 in the morning, and the animals were measured by blood sampling by tail-tip pruning without anesthesia. Fasting blood sugar. Then the animals were anesthetized with CO 2 and sacrificed. Blood was collected from the heart and the plasma was centrifuged. The plasma was used to determine plasma total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), triglycerides (TG), and insulin content ( Calculate HOMA-IR). The liver was separated for homogenization, and the supernatant of the homogenate was centrifuged to determine the content of liver triglycerides.
  • TC plasma total cholesterol
  • LDL-C low-density lipoprotein cholesterol
  • TG triglycerides
  • insulin content Calculate HOMA-IR
  • Mouse HOMA-IR [(fasting insulin(mIU/L) ⁇ fastingglucose(mmol/l)]/22.5. The result is expressed as the mean ⁇ SEM of 6 animals.

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Abstract

L'invention concerne un composé polypeptidique et son utilisation, des médicaments polypeptidiques à agonistes doubles ou triples qui activent un récepteur de peptide-1 de type glucagon et activent éventuellement un récepteur de polypeptide insulinotrope glucose-dépendant et un récepteur de glucagon, et leur utilisation pour le traitement du syndrome métabolique.
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102180963A (zh) * 2011-04-22 2011-09-14 中国药科大学 胰高血糖素样肽-1(glp-1)类似物及其应用
CN103087175A (zh) * 2012-11-30 2013-05-08 中国药科大学 新型长效化胰高血糖素样肽-1(glp-1)类似物及其应用
CN103458920A (zh) * 2010-12-22 2013-12-18 印第安那大学科技研究公司 表现出gip受体活性的胰高血糖素类似物
WO2017160669A1 (fr) * 2016-03-18 2017-09-21 Merck Sharp & Dohme Corp. Conjugués d'insuline-incrétine
WO2017189342A1 (fr) * 2016-04-26 2017-11-02 Merck Sharp & Dohme Corp. Conjugués dimères d'insuline-incrétine

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103458920A (zh) * 2010-12-22 2013-12-18 印第安那大学科技研究公司 表现出gip受体活性的胰高血糖素类似物
CN102180963A (zh) * 2011-04-22 2011-09-14 中国药科大学 胰高血糖素样肽-1(glp-1)类似物及其应用
CN103087175A (zh) * 2012-11-30 2013-05-08 中国药科大学 新型长效化胰高血糖素样肽-1(glp-1)类似物及其应用
WO2017160669A1 (fr) * 2016-03-18 2017-09-21 Merck Sharp & Dohme Corp. Conjugués d'insuline-incrétine
WO2017189342A1 (fr) * 2016-04-26 2017-11-02 Merck Sharp & Dohme Corp. Conjugués dimères d'insuline-incrétine

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
EVERS ANDREAS, BOSSART MARTIN, PFEIFFER-MAREK STEFANIA, ELVERT RALF, SCHREUDER HERMAN, KURZ MICHAEL, STENGELIN SIEGFRIED, LORENZ M: "Dual Glucagon-like Peptide 1 (GLP-1)/Glucagon Receptor Agonists Specifically Optimized for Multidose Formulations", JOURNAL OF MEDICINAL CHEMISTRY, AMERICAN CHEMICAL SOCIETY, vol. 61, no. 13, 12 July 2018 (2018-07-12), pages 5580 - 5593, XP055829008, ISSN: 0022-2623, DOI: 10.1021/acs.jmedchem.8b00292 *

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