WO2021057541A1 - 一种艾塞那肽类似物 - Google Patents

一种艾塞那肽类似物 Download PDF

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WO2021057541A1
WO2021057541A1 PCT/CN2020/115222 CN2020115222W WO2021057541A1 WO 2021057541 A1 WO2021057541 A1 WO 2021057541A1 CN 2020115222 W CN2020115222 W CN 2020115222W WO 2021057541 A1 WO2021057541 A1 WO 2021057541A1
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fmoc
glu
ser
gly
pro
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French (fr)
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周述靓
王鹏
邓岚
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Chengdu Aoda Biotechnology Co Ltd
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Chengdu Aoda Biotechnology Co Ltd
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Priority to US17/764,041 priority Critical patent/US20220347270A1/en
Priority to KR1020227013441A priority patent/KR102907806B1/ko
Priority to JP2022519514A priority patent/JP7491610B2/ja
Priority to BR112022005446A priority patent/BR112022005446A2/pt
Priority to EP20869930.6A priority patent/EP4036108A4/en
Publication of WO2021057541A1 publication Critical patent/WO2021057541A1/zh
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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
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • 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
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/14—Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • 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
    • 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
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • 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
    • C07K14/57563—Vasoactive intestinal peptide [VIP]; Related peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to an analogue of exenatide and its use.
  • the analogue is a glucagon-like peptide-1 (GLP-1) analogue.
  • GLP-1 is a secretin secreted by intestinal L cells, which can promote insulin secretion, inhibit the release of glucagon, stimulate the proliferation of islet B cells, induce the regeneration of islet B cells, prevent the apoptosis of islet B cells, and improve Insulin sensitivity and increased glucose utilization play an important role in the occurrence and development of type II diabetes.
  • GLP-1 In patients with type II diabetes, the "intestinal stimulating effect" is impaired, which is mainly manifested as the increase in GLP-1 concentration after a meal is less than that of normal people, but its effect of promoting insulin secretion and lowering blood sugar is not significantly impaired. Therefore, GLP-1 can be used as an important target for the treatment of type II diabetes. At the same time, GLP-1 is glucose-concentration-dependent. Its hypoglycemic properties are the basis and guarantee for its clinical application safety, thereby avoiding people’s concerns about existing diabetes. Therapeutic drugs and programs may cause patients to worry about severe hypoglycemia, and have broad application prospects in the field of diabetes treatment.
  • GLP-1 produced by the human body is very unstable and easily degraded by dipeptidyl peptidase IV (DPP-IV) in the body. Its plasma half-life is short, which limits Clinical application of GLP-1. At the same time, many type II diabetic patients are unwilling to take daily injections, so the development of safe and effective GLP-1 analogs that can be administered once a week has greater prospects.
  • DPP-IV dipeptidyl peptidase IV
  • the present invention provides an analog of exenatide and its use.
  • the analog is a glucagon-like peptide-1 (GLP-1) analog.
  • the present invention first provides a compound represented by structure I, a pharmaceutically acceptable salt, solvate, chelate or non-covalent complex formed by the compound, and a drug based on the compound Precursor, or any mixture of the above forms.
  • AA1 in structure I is:
  • CH 3 CH (CH 3 ) 2 , C (CH 3 ) 3 , CH (CH 2 CH 3 ) 2 , C (CH 2 CH 3 ) 3 , CH (CH 2 CH 2 CH 3 ) 2 , C ( CH 2 CH 2 CH 3 ) 3 , CH(CH(CH 3 )) 2 , C(CH(CH 3 )) 3 ,
  • AA2 in structure I is Lys, or Dah, or Orn, or Dab, or Dap;
  • AA3 in structure I is NH 2 or OH
  • R in structure I is HO 2 C(CH 2 ) n1 CO-( ⁇ Glu) n2 -(PEG n3 (CH2) n4 CO) n5-
  • n1 is an integer from 10 to 20;
  • n2 is an integer from 1 to 5;
  • n3 is an integer from 1 to 30;
  • n4 is an integer from 1 to 5;
  • n5 is an integer from 1 to 5.
  • the present invention also provides a pharmaceutical composition comprising the compound according to the present invention, and the pharmaceutical composition provided with the compound of the present invention is used for preparing a medicine for the treatment of diseases.
  • the pharmaceutical composition is used in the preparation of a medicament for the treatment of at least one of the following diseases, the diseases including type II diabetes, impaired glucose tolerance, type I diabetes, obesity, hypertension, metabolic syndrome, blood lipids Abnormalities, cognitive impairment, atherosclerosis, myocardial infarction, coronary heart disease, cardiovascular disease, stroke, inflammatory bowel syndrome and/or indigestion or gastric ulcer, liver fibrotic disease and pulmonary fibrotic disease.
  • diseases including type II diabetes, impaired glucose tolerance, type I diabetes, obesity, hypertension, metabolic syndrome, blood lipids Abnormalities, cognitive impairment, atherosclerosis, myocardial infarction, coronary heart disease, cardiovascular disease, stroke, inflammatory bowel syndrome and/or indigestion or gastric ulcer, liver fibrotic disease and pulmonary fibrotic disease.
  • the pharmaceutical composition is used in the preparation of a medicament for treating type II diabetes with delayed drug effect and/or preventing the deterioration of type II diabetes.
  • the pharmaceutical composition is used in the preparation of drugs for reducing food intake, reducing ⁇ -cell apoptosis, increasing pancreatic ⁇ -cell function, increasing ⁇ -cell mass and/or restoring glucose sensitivity to ⁇ -cells .
  • the present invention further provides a method for administering the compound to a subject to adjust blood glucose in the body.
  • any chemical structure within the scope described herein, whether part or the whole structure contains the above-mentioned similar structure includes all possible enantiomers and diastereomers of the compound, including A simple stereoisomer (such as a simple geometric isomer, a simple enantiomer or a simple diastereomer) and any mixture of these isomers.
  • the compounds of structural formula I include, but are not limited to, optical isomers, racemates and/or other mixtures of these compounds.
  • a single enantiomer or diastereomer, such as an optical isomer can be obtained by asymmetric synthesis or racemate resolution.
  • the resolution of racemates can be achieved by different methods, such as conventional recrystallization with reagents that assist resolution, or chromatographic methods.
  • the compounds of structural formula I also contain cis and/or trans isomers with double bonds.
  • the compounds of the present invention include, but are not limited to, the compounds represented by structural formula I and all of their pharmaceutically usable different forms.
  • the pharmaceutically usable different forms of these compounds include various pharmaceutically acceptable salts, solvates, complexes, chelates, non-covalent complexes, prodrugs based on the above-mentioned substances and the above-mentioned forms. Any mixture.
  • the compound shown in structure I provided by the present invention has stable properties, is not easily degraded by dipeptidyl peptidase IV (DPP-IV) in the body, and is a long-acting GLP-I analog with a significant hypoglycemic effect.
  • DPP-IV dipeptidyl peptidase IV
  • the present invention discloses a glucagon-like peptide-1 (GLP-1) analogue and its use.
  • GLP-1 glucagon-like peptide-1
  • Those skilled in the art can learn from the content of this article and appropriately improve relevant parameters to achieve it.
  • all similar replacements and modifications are obvious to those skilled in the art, and they are all deemed to be included in the present invention.
  • the method of the present invention has been described through preferred embodiments, and the relevant personnel can obviously modify or appropriately change and combine the compounds and preparation methods described herein without departing from the content, spirit and scope of the present invention to achieve and Apply the technology of the present invention.
  • the preparation method includes: preparing the peptide resin by solid-phase peptide synthesis, then acid hydrolyzing the peptide resin to obtain the crude product, and finally the crude product is purified to obtain the pure product; wherein the step of preparing the peptide resin by the solid-phase peptide synthesis method is to solidify the peptide resin on the carrier resin.
  • the phase coupling synthesis method sequentially connects the corresponding protected amino acids or fragments in the following sequences to prepare peptide resins:
  • the amount of the Fmoc-protected amino acid or protected amino acid fragment is 1.2-6 times the total moles of the resin charged; preferably 2.5-3.5 times.
  • the substitution value of the carrier resin is 0.2-1.0 mmol/g resin, and the preferred substitution value is 0.3-0.5 mmol/g resin.
  • the solid-phase coupling synthesis method is: the protected amino acid-resin obtained in the previous step reaction removes the Fmoc protective group and then couples with the next protected amino acid.
  • the deprotection time for Fmoc protection is 10-60 minutes, preferably 15-25 minutes.
  • the coupling reaction time is 60-300 minutes, preferably 100-140 minutes.
  • the coupling reaction requires the addition of a condensation reagent, which is selected from DIC (N,N-diisopropylcarbodiimide), N,N-dicyclohexylcarbodiimide, and benzotriazole hexafluorophosphate -1-yl-oxytripyrrolidinyl phosphorus, 2-(7-aza-1H-benzotriazol-1-yl)-1,1,3,3-tetramethylurea hexafluorophosphate , Benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate or O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoro
  • a condensation reagent which is selected from DIC (N,N-diisopropylcarbodiimide), N,N-dicyclohexylcarbodiimide, and benzotriazole hexaflu
  • the coupling reaction needs to add an activating reagent, and the activating reagent is selected from 1-hydroxybenzotriazole or N-hydroxy-7-azabenzotriazole, preferably 1-hydroxybenzotriazole.
  • the amount of the activating reagent is 1.2-6 times the total moles of amino groups in the amino resin, preferably 2.5-3.5 times.
  • the reagent for removing Fmoc protection is a PIP/DMF (piperidine/N,N-dimethylformamide) mixed solution, and the mixed solution contains 10-30% piperidine (V ).
  • the amount of the de-Fmoc protection reagent is 5-15 mL per gram of amino resin, preferably 8-12 mL per gram of amino resin.
  • the peptide resin undergoes acid hydrolysis to simultaneously remove the resin and side chain protecting groups to obtain a crude product:
  • the acid hydrolyzing agent used in the acid hydrolysis of the peptide resin is a mixed solvent of trifluoroacetic acid (TFA), 1,2-ethanedithiol (EDT) and water, and the volume ratio of the mixed solvent is: TFA It is 80-95%, EDT is 1-10%, and the balance is water.
  • the volume ratio of the mixed solvent is as follows: TFA is 89% to 91%, EDT is 4% to 6%, and the balance is water. Optimally, the volume ratio of the mixed solvent is: TFA is 90%, EDT is 5%, and the balance is water.
  • the dosage of the acid hydrolyzing agent is 4-15 mL of acid hydrolyzing agent per gram of peptide resin; preferably, 7-10 mL of acid hydrolyzing agent is required per gram of peptide resin.
  • the cleavage time using an acid hydrolyzing agent is 1 to 6 hours at room temperature, preferably 3 to 4 hours.
  • the crude product is purified by high performance liquid chromatography and freeze-dried to obtain a pure product.
  • the peptide resin is prepared by sequentially coupling with the protected amino acids shown in the following table.
  • the protected amino acids corresponding to the protected amino acids used in this example are as follows:
  • n Protected amino acids 1 Fmoc-Lys(Alloc) 2 Fmoc-Ser(tBu) 3 Fmoc-Pro 4 Fmoc-Pro 5 Fmoc-Pro 6 Fmoc-Ala 7 Fmoc-Gly 8 Fmoc-Ser(tBu) 9 Fmoc-Ser(tBu) 10 Fmoc-Pro 11 Fmoc-Gly 12 Fmoc-Gly 13 Fmoc-Asn(Trt) 14 Fmoc-Lys(Boc) 15 Fmoc-Leu 16 Fmoc-Trp(Boc) 17 Fmoc-Glu(OtBu) 18 Fmoc-Ile 19 Fmoc-Phe 20 Fmoc-Leu twenty one Fmoc-Arg(pbf) twenty two Fmoc-Val
  • Fmoc-Ala twenty three Fmoc-Ala twenty four Fmoc-Glu(OtBu) 25 Fmoc-Glu(OtBu) 26 Fmoc-Glu(OtBu) 27 Fmoc-Met 28 Fmoc-Gln(Trt) 29 Fmoc-Lys(Boc) 30 Fmoc-Ser(tBu) 31 Fmoc-Leu 32 Fmoc-Asp(OtBu) 33 Fmoc-Ser(tBu) 34 Fmoc-Thr(tBu) 35 Fmoc-Phe 36 Fmoc-Thr(tBu) 37 Fmoc-Gly 38 Fmoc-Glu(OtBu) 39 Fmoc-Dhthr 40 Boc-His(Trt) Side chain-1 Fmoc-AEEA Side chain-2 Fmoc-AEEA Side chain-3 Fmoc- ⁇ Glu-OtBu Side chain-4 18 alkanedioic acid mono-
  • the activated first protected amino acid solution is added to the Fmoc-free resin, the coupling is reacted for 60-300 minutes, and filtered and washed to obtain a resin containing 1 protected amino acid.
  • Purification was carried out by high performance liquid chromatography.
  • the chromatographic packing used for purification was 10 ⁇ m reversed-phase C18, the mobile phase system was 0.1% TFA/water solution-0.1% TFA/acetonitrile solution, and the flow rate of the 30mm*250mm column was 20mL/min.
  • Gradient system elution, circular injection purification take the crude solution and load it on the chromatographic column, start the mobile phase elution, collect the main peak and evaporate the acetonitrile to obtain the purified intermediate concentrate;
  • the purified intermediate concentrate was filtered with a 0.45 ⁇ m filter membrane for use, and the salt was replaced by high performance liquid chromatography.
  • the mobile phase system was 1% acetic acid/water-acetonitrile, and the chromatographic packing for purification was 10 ⁇ m reversed-phase C18, 30mm*250mm
  • the flow rate of the chromatographic column is 20mL/min (the corresponding flow rate can be adjusted according to the different specifications of the chromatographic column); using gradient elution, loop loading method, load the sample on the chromatographic column, start the mobile phase elution, collect the spectrum, and observe For the change of absorbance, collect the main peak of salt exchange and check the purity with the analytical liquid phase.
  • the preparation method is the same as in Example 1.
  • the protected amino acids used are as follows:
  • n Protected amino acids 1 Fmoc-Lys(Alloc) 2 Fmoc-Ser(tBu) 3 Fmoc-Pro 4 Fmoc-Pro 5 Fmoc-Pro 6 Fmoc-Ala 7 Fmoc-Gly 8 Fmoc-Ser(tBu) 9 Fmoc-Ser(tBu)
  • Fmoc-Pro 11 Fmoc-Gly 12 Fmoc-Gly 13 Fmoc-Asn(Trt) 14 Fmoc-Lys(Boc) 15 Fmoc-Leu 16 Fmoc-Trp(Boc) 17 Fmoc-Glu(OtBu) 18 Fmoc-Ile 19 Fmoc-Phe 20 Fmoc-Leu twenty one Fmoc-Arg(pbf) twenty two Fmoc-Val twenty three Fmoc-Ala twenty four Fmoc-Glu(OtBu) 25 Fmoc-Glu(OtBu) 26 Fmoc-Glu(OtBu) 27 Fmoc-Met 28 Fmoc-Gln(Trt) 29 Fmoc-Lys(Boc) 30 Fmoc-Ser(tBu) 31 Fmoc-Leu 32 Fmoc-Asp(OtBu) 33 Fmoc-Ser(t
  • the pure product was 6.2g, the purity was 95.8%, and the total yield was 12.2%.
  • the molecular weight is 5070.6 (100% M+H).
  • the preparation method is the same as in Example 1.
  • the protected amino acids used are as follows:
  • n Protected amino acids 1 Fmoc-Lys(Alloc) 2 Fmoc-Ser(tBu) 3 Fmoc-Pro 4 Fmoc-Pro 5 Fmoc-Pro 6 Fmoc-Ala 7 Fmoc-Gly 8 Fmoc-Ser(tBu) 9 Fmoc-Ser(tBu)
  • Fmoc-Pro 11 Fmoc-Gly 12 Fmoc-Gly 13 Fmoc-Asn(Trt) 14 Fmoc-Lys(Boc) 15 Fmoc-Leu 16 Fmoc-Trp(Boc) 17 Fmoc-Glu(OtBu) 18 Fmoc-Ile 19 Fmoc-Phe 20 Fmoc-Leu twenty one Fmoc-Arg(pbf) twenty two Fmoc-Val twenty three Fmoc-Ala twenty four Fmoc-Glu(OtBu) 25 Fmoc-Glu(OtBu) 26 Fmoc-Glu(OtBu) 27 Fmoc-Met 28 Fmoc-Gln(Trt) 29 Fmoc-Lys(Boc) 30 Fmoc-Ser(tBu) 31 Fmoc-Leu 32 Fmoc-Asp(OtBu) 33 Fmoc-Ser(t
  • the pure product was 8.9 g, the purity was 98.5%, and the total yield was 17.6%.
  • the molecular weight is 5043.2 (100% M+H).
  • the preparation method is the same as in Example 1.
  • the protected amino acids used are as follows:
  • n Protected amino acids 1 Fmoc-Lys(Alloc) 2 Fmoc-Ser(tBu) 3 Fmoc-Pro 4 Fmoc-Pro 5 Fmoc-Pro 6 Fmoc-Ala 7 Fmoc-Gly 8 Fmoc-Ser(tBu) 9 Fmoc-Ser(tBu) 10 Fmoc-Pro
  • the preparation method is the same as in Example 1.
  • the protected amino acids used are as follows:
  • n Protected amino acids 1 Fmoc-Lys(Alloc) 2 Fmoc-Ser(tBu) 3 Fmoc-Pro 4 Fmoc-Pro 5 Fmoc-Pro 6 Fmoc-Ala 7 Fmoc-Gly 8 Fmoc-Ser(tBu) 9 Fmoc-Ser(tBu) 10 Fmoc-Pro 11 Fmoc-Gly
  • Fmoc-Gly 13 Fmoc-Asn(Trt) 14 Fmoc-Lys(Boc) 15 Fmoc-Leu 16 Fmoc-Trp(Boc) 17 Fmoc-Glu(OtBu) 18 Fmoc-Ile 19 Fmoc-Phe 20 Fmoc-Leu twenty one Fmoc-Arg(pbf) twenty two Fmoc-Val twenty three Fmoc-Ala twenty four Fmoc-Glu(OtBu) 25 Fmoc-Glu(OtBu) 26 Fmoc-Glu(OtBu) 27 Fmoc-Met 28 Fmoc-Gln(Trt) 29 Fmoc-Lys(Boc) 30 Fmoc-Ser(tBu) 31 Fmoc-Leu 32 Fmoc-Asp(OtBu) 33 Fmoc-Ser(tBu) 34 Fmoc-Thr(tBu) 35 F
  • the pure product was 5.6 g, the purity was 97.6%, and the total yield was 10.6%.
  • the molecular weight is 5263.8 (100% M+H).
  • the preparation method is the same as in Example 1.
  • the protected amino acids used are as follows:
  • n Protected amino acids 1 Fmoc-Lys(Alloc) 2 Fmoc-Ser(tBu) 3 Fmoc-Pro 4 Fmoc-Pro 5 Fmoc-Pro 6 Fmoc-Ala 7 Fmoc-Gly 8 Fmoc-Ser(tBu) 9 Fmoc-Ser(tBu) 10 Fmoc-Pro 11 Fmoc-Gly 12 Fmoc-Gly
  • GLP-1R mainly exists on the surface of pancreatic ⁇ cells and is a G protein-coupled receptor (GPCRs). Under the stimulation of its specific agonist, GLP-1R can activate the adenylate cyclase pathway in the cell, increase the level of cAMP, and ultimately lead to the production and release of insulin.
  • the cell line stably transfected with GLP-1R was stimulated by the test substance to rapidly increase the intracellular cAMP level.
  • the relative light unit (RLU) after each dose of stimulated cells was measured by chemiluminescence method, and then the agonist EC50, this activity determination method is currently a universal GLP-1 receptor agonist activity detection method at home and abroad.
  • CHO-K1 cell line stably expressing GLP-1R, stimulated the stable cells with different concentrations of agonist, and measured the relative light units of the cells after stimulation at each dose to obtain the biological activity of the agonist.
  • Each compound was divided into two administration groups: SD rats, 4 males in each group, a total of 8 rats.
  • Tail vein intravenous injection group The dose is 1 mg/kg, and the blood is taken from the rat's orbital vein before the drug (0h) and 30min, 1h, 2h, 4h, 8h, 24h, 48h, 96h, 144h after the drug, and centrifuged. Plasma sample.
  • Subcutaneous administration group the dose is 1 mg/kg, and the blood is collected from the orbital vein of the rat before (0h) and 1h, 2h, 3h, 4h, 8h, 24h, 48h, 96h, 144h after administration, and the plasma is separated by centrifugation sample.
  • the plasma concentrations of the corresponding compounds in the plasma samples of SD rats were determined by LC/MS. After intravenous and subcutaneous administration, the half-life of the compound in SD rats subcutaneous (SC) administration is shown in the following table:

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Abstract

涉及医药合成领域，公开了一种艾塞那肽类似物。所述艾塞那肽类似物，用于制备治疗疾病的药物组合物，所述药物组合物在制备治疗下述至少一种疾病的药物中的用途，所述疾病包括II型糖尿病、糖耐量受损、I型糖尿病、肥胖、高血压、代谢综合征、血脂异常、认知障碍、动脉粥样硬化、心肌梗塞、冠状动脉心脏病、心血管疾病、中风、炎性肠道综合征和/或消化不良或胃溃疡、肝纤维化疾病和肺纤维化疾病。

Description

一种艾塞那肽类似物
本申请要求于2019年09月25日提交中国专利局、申请号为201910908468.3、发明名称为“一种艾塞那肽类似物”的中国专利申请的优先权，其全部内容通过引用结合在本申请中。
技术领域
本发明涉及一种艾塞那肽类似物及其用途，该类似物是一种胰高血糖素样肽-1(GLP-1)类似物。
背景技术
糖尿病已成为继心脑血管疾病和肿瘤后排名第三的非传染性疾病，世界卫生组织(WHO)预测：2030年全世界糖尿病患者将超过3.6亿，其中90％以上为II型糖尿病。GLP-1是一种由肠道L细胞分泌的促胰素，具有促进胰岛素分泌、抑制胰高血糖素的释放、刺激胰岛B细胞增殖、诱导胰岛B细胞再生、阻止胰岛B细胞凋亡、改善胰岛素敏感性和增加葡萄糖的利用等作用，在II型糖尿病的发生、发展中起着重要作用。II型糖尿病患者，其“肠促胰岛效应”受损，主要表现为进餐后GLP-1浓度升高幅度较正常人有所减少，但其促进胰岛素分泌以及降血糖的作用并无明显受损，因此GLP-1可以作为II型糖尿病治疗的一个重要靶点，同时GLP-1具有葡萄糖浓度依赖性，其降糖特性是在其临床应用安全性的基础与保障，从而免除了人们对现有糖尿病治疗药物及方案可能造成患者严重低血糖的担心，在糖尿病治疗领域中具有广阔的应用前景。
但GLP-1应用于临床也面临着巨大问题，人体自身产生的GLP-1很不稳定，极易被体内的二肽基肽酶IV(DPP-IV)降解，其血浆半衰期较短，限制了GLP-1的临床应用。同时许多II型糖尿病患者不愿意每天注射给药，所以开发能够每周用药一次的安全有效的GLP-1类似物具有更大的前景。
发明内容
本发明提供了一种艾塞那肽类似物及其用途，该类似物是一种胰高血 糖素样肽-1(GLP-1)类似物。
为实现上述目的，本发明首先提供了一种结构I所示的化合物，该化合物所成的可药用的盐、溶剂化物、螯合物或非共价复合物，基于该化合物基础上的药物前体，或上述形式的任意混合物。
AA1-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met-Glu-Glu-Glu-Ala-Val-Arg-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-AA2(R)-AA3
结构I
结构I中的AA1为:
Figure PCTCN2020115222-appb-000001
其中X 1和X 2为H，
或为CH 3、CH(CH 3) 2、C(CH 3) 3、CH(CH 2CH 3) 2、C(CH 2CH 3) 3、CH(CH 2CH 2CH 3) 2、C(CH 2CH 2CH 3) 3、CH(CH(CH 3)) 2、C(CH(CH 3)) 3、
或为CH 2CH 3、CH 2CH(CH 3) 2、CH 2C(CH 3) 3、CH 2CH(CH 2CH 3) 2、CH 2C(CH 2CH 3) 3、CH 2CH(CH 2CH 2CH 3) 2、CH 2C(CH 2CH 2CH 3) 3、CH 2CH(CH(CH 3)) 2、CH 2C(CH(CH 3)) 3、
结构I中的AA2为Lys，或为Dah，或为Orn，或为Dab，或为Dap；
结构I中的AA3为NH 2，或为OH；
结构I中的R为HO 2C(CH 2) n1CO-(γGlu) n2-(PEG n3(CH2) n4CO) n5-
其中：n1为10至20的整数；
n2为1至5的整数；
n3为1至30的整数；
n4为1至5的整数；
n5为1至5的整数。
本发明还提供了包括根据本发明化合物的药物组合物，以及提供了本 发明化合物的药物组合物用于制备治疗疾病的药物用途。
作为优选，所述药物组合物在制备治疗下述至少一种疾病的药物中的用途，所述疾病包括II型糖尿病、糖耐量受损、I型糖尿病、肥胖、高血压、代谢综合征、血脂异常、认知障碍、动脉粥样硬化、心肌梗塞、冠状动脉心脏病、心血管疾病、中风、炎性肠道综合征和/或消化不良或胃溃疡、肝纤维化疾病和肺纤维化疾病。
作为优选，所述药物组合物在制备治疗II型糖尿病药效延迟和/或预防II型糖尿病恶化的药物中的应用。
作为优选，所述药物组合物在制备减少食物摄入量、减少β细胞凋亡、增加胰岛β细胞功能、增加β-细胞团和/或回复葡萄糖对β-细胞的敏感性的药物中的应用。
本发明还进一步提供了对治疗对象施用所述化合物以调节体内血糖的方法。
本发明所涉及到的更多内容在以下有详细描述，或者有些也可以在本发明的实施例中体会。
除非另有所指，本文中所用来表示不同成分的数量、反应条件，在任意情况下都可解读为“大致的”、“大约的”意思。相应的，除有明确的特指外，在下述以及权利要求中所引用的数字参数都是大致的参数，在各自的实验条件下由于标准误差的不同，有可能会得到不同的数字参数。
本文中，当一个化合物的化学结构式和化学名称有分歧或疑义时，以化学结构式确切定义此化合物。本文所描述的化合物有可能含有一个或多个手性中心，和/或者双键以及诸如此类的结构，也可能存在立体异构体，包括双键的异构体(比如几何异构体)、旋光对映异构体或者非对映异构体。相应的，在本文描述范围内的任意化学结构，无论是部分或整体结构中含有上述类似结构，都包括了此化合物的所有可能的对映异构体和非对映异构体，其中也包括了单纯的任一种立体异构体(如单纯的几何异构体、单纯的对映异构体或者单纯的非对映异构体)以及这些异构体的任意一种混合物。这些消旋异构体和立体异构体的混合物由本领域技术人员利用不停的分离技术或手性分子合成的方法也可进一步被拆分成其组成成分的 对映异构体或立体异构体。
结构式I的化合物包含了，但并不仅限于，这些化合物的光学异构体、消旋体和/或其他的混合物。上述情况下，其中单一的对映异构体或非对映异构体，如有旋光的异构体，可以用不对称合成的方法或消旋体拆分的方法获得。消旋体的拆分可用不同的方法实现，如常规的用助拆分的试剂重结晶，或用色谱方法。另外，结构式I的化合物也包含了带双键的顺式和/或反式的异构体。
本发明所述化合物包含但不限于，结构式I所示化合物以及他们所有的在药学上可用的不同形式。这些化合物的药学上可用的不同形式包括各种可药用的盐、溶剂化物、络合物、螯合物、非共价的复合物、基于上述物质基础上的药物前体和上述这些形式的任意混合物。
本发明提供的结构I所示的化合物性质稳定，不易被体内的二肽基肽酶IV(DPP-IV)降解，为一种长效的GLP-I类似物，具有显著的降糖效果。
具体实施方式
本发明公开了一种胰高血糖素样肽-1(GLP-1)类似物及其用途，本领域技术人员可以借鉴本文内容，适当改进相关参数实现。特别需要指出的是，所有类似的替换和改动对本领域技术人员来说是显而易见的，它们都被视为包括在本发明。本发明的方法已经通过较佳实施例进行了描述，相关人员明显能在不脱离本发明内容、精神和范围内对本文所述的的化合物和制备方法进行改动或适当变更与组合，来实现和应用本发明技术。
本发明中涉及的英文缩写所对应的中文名称见下表所示：
Figure PCTCN2020115222-appb-000002
Figure PCTCN2020115222-appb-000003
实施例1化合物1的制备
Figure PCTCN2020115222-appb-000004
制备方法，包括：采用固相多肽合成法制备肽树脂，肽树脂再经酸解得到粗品，最后粗品经过纯化得到纯品；其中固相多肽合成法制备肽树脂的步骤为在载体树脂上通过固相偶联合成法依次接入下列序列中相对应的保护氨基酸或片段，制备肽树脂：
上述制备方法中，所述的Fmoc-保护氨基酸或保护氨基酸片段的用量 为所投料树脂总摩尔数的1.2～6倍；优选为2.5～3.5倍。
上述制备方法中，所述的载体树脂取代值为0.2～1.0mmol/g树脂，优选的取代值为0.3～0.5mmol/g树脂。
作为本发明优选的方案，所述固相偶联合成法为：前一步反应得到的保护氨基酸-树脂脱去Fmoc保护基后再与下一个保护氨基酸偶联反应。所述的去Fmoc保护的脱保护时间为10～60分钟，优选的为15～25分钟。所述的偶联反应时间为60～300分钟，优选的为100～140分钟。
所述的偶联反应需添加缩合试剂，缩合试剂选自DIC(N,N-二异丙基碳二亚胺)、N,N-二环己基碳二亚胺，六氟磷酸苯并三唑-1-基-氧基三吡咯烷基磷、2-(7-氮杂-1H-苯并三氮唑-1-基)-1,1,3,3-四甲基脲六氟磷酸酯、苯并三氮唑-N,N,N',N'-四甲基脲六氟磷酸盐或O-苯并三氮唑-N,N,N',N'-四甲基脲四氟硼酸酯中的一种；优选的为N,N-二异丙基碳二亚胺。所述缩合试剂的摩尔用量为氨基树脂中氨基总摩尔数的1.2～6倍，优选为2.5～3.5倍。
所述的偶联反应需添加活化试剂，活化试剂选自1-羟基苯并三唑或N-羟基-7-氮杂苯并三氮唑，优选的为1-羟基苯并三唑。活化试剂的用量为氨基树脂中氨基总摩尔数的1.2～6倍，优选的为2.5～3.5倍。
作为本发明优选的方案，所述的脱去Fmoc保护的试剂为PIP/DMF(哌啶/N,N-二甲基甲酰胺)混合溶液，混合溶液中含哌啶为10～30％(V)。去Fmoc保护试剂的用量为每克氨基树脂5～15mL，优选的为每克氨基树脂8～12mL。
优选的，肽树脂经酸解同时脱去树脂及侧链保护基得到粗品：
进一步优选的，所述肽树脂酸解时采用的酸解剂为三氟醋酸(TFA)、1,2-乙二硫醇(EDT)和水的混合溶剂，混合溶剂的体积配比为：TFA为80～95％，EDT为1～10％，余量为水。
更进一步优选的，混合溶剂的体积配比为：TFA为89～91％、EDT为4～6％，余量为水。最优的，混合溶剂的体积配比为：TFA为90％、EDT为5％，余量为水。
所述酸解剂用量为每克肽树脂需要4～15mL酸解剂；优选的，每克 肽树脂需要7～10mL酸解剂。
使用酸解剂裂解的时间为室温条件下1～6小时，优选的为3～4小时。
进一步的，粗品经高效液相色谱纯化、冻干得到纯品。
1、肽树脂的合成
使用Rink Amide BHHA树脂为载体树脂，通过去Fmoc保护和偶联反应，依次与下表所示的保护氨基酸偶联，制得肽树脂。本实施例使用的保护氨基酸相对应的保护氨基酸如下所示：
接肽顺序n＝ 保护氨基酸
1 Fmoc-Lys(Alloc)
2 Fmoc-Ser(tBu)
3 Fmoc-Pro
4 Fmoc-Pro
5 Fmoc-Pro
6 Fmoc-Ala
7 Fmoc-Gly
8 Fmoc-Ser(tBu)
9 Fmoc-Ser(tBu)
10 Fmoc-Pro
11 Fmoc-Gly
12 Fmoc-Gly
13 Fmoc-Asn(Trt)
14 Fmoc-Lys(Boc)
15 Fmoc-Leu
16 Fmoc-Trp(Boc)
17 Fmoc-Glu(OtBu)
18 Fmoc-Ile
19 Fmoc-Phe
20 Fmoc-Leu
21 Fmoc-Arg(pbf)
22 Fmoc-Val
23 Fmoc-Ala
24 Fmoc-Glu(OtBu)
25 Fmoc-Glu(OtBu)
26 Fmoc-Glu(OtBu)
27 Fmoc-Met
28 Fmoc-Gln(Trt)
29 Fmoc-Lys(Boc)
30 Fmoc-Ser(tBu)
31 Fmoc-Leu
32 Fmoc-Asp(OtBu)
33 Fmoc-Ser(tBu)
34 Fmoc-Thr(tBu)
35 Fmoc-Phe
36 Fmoc-Thr(tBu)
37 Fmoc-Gly
38 Fmoc-Glu(OtBu)
39 Fmoc-Dhthr
40 Boc-His(Trt)
侧链-1 Fmoc-AEEA
侧链-2 Fmoc-AEEA
侧链-3 Fmoc-γGlu-OtBu
侧链-4 18烷二酸单叔丁酯
备注 Dhthr：脱羟基苏氨酸
(1)接入主链第1个保护氨基酸
取0.03mol第1个保护氨基酸和0.03mol HOBt，用适量DMF溶解；另取0.03mol DIC，搅拌下慢慢加入至保护氨基酸DMF溶液中，于室温环境中搅拌反应30分钟，得到活化后的保护氨基酸溶液，备用。
取0.01mol的Rink amide MBHA树脂(取代值约0.4mmol/g)，采用20％PIP/DMF溶液去保护25分钟，洗涤过滤得到去Fmoc的树脂。
将活化后的第1个保护氨基酸溶液加入到已去Fmoc的树脂中，偶联 反应60～300分钟，过滤洗涤，得含1个保护氨基酸的树脂。
(2)接入主链第2～40个保护氨基酸
采用上述接入主链第1个保护氨基酸同样方法，依次接入上述对应的第2～40个保护氨基酸，得含主链40个氨基酸的树脂。
(3)接入侧链第1个保护氨基酸
取0.03mol侧链第1个保护氨基酸和0.03mol HOBt，用适量DMF溶解；另取0.03mol DIC，搅拌下慢慢加入至保护氨基酸DMF溶液中，于室温环境中搅拌反应30分钟，得到活化后的保护氨基酸溶液。
取2.5mmol四三苯基膦钯和25mmol苯硅烷，用适量二氯甲烷溶解，去保护4小时，过滤洗涤，得到去Alloc的树脂备用。
将加入活化后的侧链第1个保护氨基酸液加入到已去Alloc的树脂，偶联反应60～300分钟，过滤洗涤，得含侧链第1个保护氨基酸的树脂。
(4)接入侧链第2～4个保护氨基酸
采用上述接入主链第1个保护氨基酸同样方法，依次接入侧链对应的第2～4个保护氨基酸和单保护脂肪酸，得到肽树脂。
2、粗品的制备
取上述肽树脂，加入体积比为TFA︰水︰EDT＝95︰5︰5的裂解试剂(裂解试剂10mL/克树脂)，搅拌均匀，室温搅拌反应3小时，反应混合物使用砂芯漏斗过滤，收集滤液，树脂再用少量TFA洗涤3次，合并滤液后减压浓缩，加入无水乙醚沉淀，再用无水乙醚洗沉淀3次，抽干得类白色粉末即为粗品。
3、纯品的制备
取上述粗品，加水搅拌，用氨水调pH8.0至完全溶解，溶液用0.45μm混合微孔滤膜过滤，纯化备用；
采用高效液相色谱法进行纯化，纯化用色谱填料为10μm的反相C18，流动相系统为0.1％TFA/水溶液-0.1％TFA/乙腈溶液，30mm*250mm的色谱柱流速为20mL/min，采用梯度系统洗脱，循环进样纯化，取粗品溶液上样于色谱柱中，启动流动相洗脱，收集主峰蒸去乙腈后，得纯化中间体浓缩液；
纯化中间体浓缩液用0.45μm滤膜滤过备用，采用高效液相色谱法进行换盐，流动相系统为1％醋酸/水溶液-乙腈，纯化用色谱填料为10μm的反相C18，30mm*250mm的色谱柱流速为20mL/min(可根据不同规格的色谱柱，调整相应的流速)；采用梯度洗脱，循环上样方法，上样于色谱柱中，启动流动相洗脱，采集图谱，观测吸收度的变化，收集换盐主峰并用分析液相检测纯度，合并换盐主峰溶液，减压浓缩，得到纯品醋酸水溶液，冷冻干燥，得纯品7.7g，纯度为95.8％，总收率为15.2％。分子量为5056.2(100％M+H)。
实施例2化合物2的制备
Figure PCTCN2020115222-appb-000005
制备方法同实施例1，使用的保护氨基酸如下表:
接肽顺序n＝ 保护氨基酸
1 Fmoc-Lys(Alloc)
2 Fmoc-Ser(tBu)
3 Fmoc-Pro
4 Fmoc-Pro
5 Fmoc-Pro
6 Fmoc-Ala
7 Fmoc-Gly
8 Fmoc-Ser(tBu)
9 Fmoc-Ser(tBu)
10 Fmoc-Pro
11 Fmoc-Gly
12 Fmoc-Gly
13 Fmoc-Asn(Trt)
14 Fmoc-Lys(Boc)
15 Fmoc-Leu
16 Fmoc-Trp(Boc)
17 Fmoc-Glu(OtBu)
18 Fmoc-Ile
19 Fmoc-Phe
20 Fmoc-Leu
21 Fmoc-Arg(pbf)
22 Fmoc-Val
23 Fmoc-Ala
24 Fmoc-Glu(OtBu)
25 Fmoc-Glu(OtBu)
26 Fmoc-Glu(OtBu)
27 Fmoc-Met
28 Fmoc-Gln(Trt)
29 Fmoc-Lys(Boc)
30 Fmoc-Ser(tBu)
31 Fmoc-Leu
32 Fmoc-Asp(OtBu)
33 Fmoc-Ser(tBu)
34 Fmoc-Thr(tBu)
35 Fmoc-Phe
36 Fmoc-Thr(tBu)
37 Fmoc-Gly
38 Fmoc-Glu(OtBu)
39 Fmoc-Dhval
40 Boc-His(Trt)
侧链-1 Fmoc-AEEA
侧链-2 Fmoc-AEEA
侧链-3 Fmoc-γGlu-OtBu
侧链-4 18烷二酸单叔丁酯
备注 Dhval：2,3-二脱氢缬氨酸
得纯品6.2g，纯度为95.8％，总收率为12.2％。分子量为5070.6(100％M+H)。
实施例3化合物3的制备
Figure PCTCN2020115222-appb-000006
制备方法同实施例1，使用的保护氨基酸如下表:
接肽顺序n＝ 保护氨基酸
1 Fmoc-Lys(Alloc)
2 Fmoc-Ser(tBu)
3 Fmoc-Pro
4 Fmoc-Pro
5 Fmoc-Pro
6 Fmoc-Ala
7 Fmoc-Gly
8 Fmoc-Ser(tBu)
9 Fmoc-Ser(tBu)
10 Fmoc-Pro
11 Fmoc-Gly
12 Fmoc-Gly
13 Fmoc-Asn(Trt)
14 Fmoc-Lys(Boc)
15 Fmoc-Leu
16 Fmoc-Trp(Boc)
17 Fmoc-Glu(OtBu)
18 Fmoc-Ile
19 Fmoc-Phe
20 Fmoc-Leu
21 Fmoc-Arg(pbf)
22 Fmoc-Val
23 Fmoc-Ala
24 Fmoc-Glu(OtBu)
25 Fmoc-Glu(OtBu)
26 Fmoc-Glu(OtBu)
27 Fmoc-Met
28 Fmoc-Gln(Trt)
29 Fmoc-Lys(Boc)
30 Fmoc-Ser(tBu)
31 Fmoc-Leu
32 Fmoc-Asp(OtBu)
33 Fmoc-Ser(tBu)
34 Fmoc-Thr(tBu)
35 Fmoc-Phe
36 Fmoc-Thr(tBu)
37 Fmoc-Gly
38 Fmoc-Glu(OtBu)
39 Fmoc-Dhthr
40 Boc-His(Trt)
侧链-1 Fmoc-PEG 5CH 2COOH
侧链-3 Fmoc-γGlu-OtBu
侧链-3 18烷二酸单叔丁酯
备注 Dhthr：脱羟基苏氨酸
得纯品8.9g，纯度为98.5％，总收率为17.6％。分子量为5043.2(100％M+H)。
实施例4化合物4的制备
Figure PCTCN2020115222-appb-000007
制备方法同实施例1，使用的保护氨基酸如下表:
接肽顺序n＝ 保护氨基酸
1 Fmoc-Lys(Alloc)
2 Fmoc-Ser(tBu)
3 Fmoc-Pro
4 Fmoc-Pro
5 Fmoc-Pro
6 Fmoc-Ala
7 Fmoc-Gly
8 Fmoc-Ser(tBu)
9 Fmoc-Ser(tBu)
10 Fmoc-Pro
11 Fmoc-Gly
12 Fmoc-Gly
13 Fmoc-Asn(Trt)
14 Fmoc-Lys(Boc)
15 Fmoc-Leu
16 Fmoc-Trp(Boc)
17 Fmoc-Glu(OtBu)
18 Fmoc-Ile
19 Fmoc-Phe
20 Fmoc-Leu
21 Fmoc-Arg(pbf)
22 Fmoc-Val
23 Fmoc-Ala
24 Fmoc-Glu(OtBu)
25 Fmoc-Glu(OtBu)
26 Fmoc-Glu(OtBu)
27 Fmoc-Met
28 Fmoc-Gln(Trt)
29 Fmoc-Lys(Boc)
30 Fmoc-Ser(tBu)
31 Fmoc-Leu
32 Fmoc-Asp(OtBu)
33 Fmoc-Ser(tBu)
34 Fmoc-Thr(tBu)
35 Fmoc-Phe
36 Fmoc-Thr(tBu)
37 Fmoc-Gly
38 Fmoc-Glu(OtBu)
39 Fmoc-Dhval
40 Boc-His(Trt)
侧链-1 Fmoc-PEG 5CH 2COOH
侧链-2 Fmoc-γGlu-OtBu
侧链-3 18烷二酸单叔丁酯
备注 Dhval：2,3-二脱氢缬氨酸
得纯品5.2g，纯度为96.3％，总收率为10.3％。分子量为5057.6(100％M+H)。
实施例5化合物5的制备
Figure PCTCN2020115222-appb-000008
制备方法同实施例1，使用的保护氨基酸如下表:
接肽顺序n＝ 保护氨基酸
1 Fmoc-Lys(Alloc)
2 Fmoc-Ser(tBu)
3 Fmoc-Pro
4 Fmoc-Pro
5 Fmoc-Pro
6 Fmoc-Ala
7 Fmoc-Gly
8 Fmoc-Ser(tBu)
9 Fmoc-Ser(tBu)
10 Fmoc-Pro
11 Fmoc-Gly
12 Fmoc-Gly
13 Fmoc-Asn(Trt)
14 Fmoc-Lys(Boc)
15 Fmoc-Leu
16 Fmoc-Trp(Boc)
17 Fmoc-Glu(OtBu)
18 Fmoc-Ile
19 Fmoc-Phe
20 Fmoc-Leu
21 Fmoc-Arg(pbf)
22 Fmoc-Val
23 Fmoc-Ala
24 Fmoc-Glu(OtBu)
25 Fmoc-Glu(OtBu)
26 Fmoc-Glu(OtBu)
27 Fmoc-Met
28 Fmoc-Gln(Trt)
29 Fmoc-Lys(Boc)
30 Fmoc-Ser(tBu)
31 Fmoc-Leu
32 Fmoc-Asp(OtBu)
33 Fmoc-Ser(tBu)
34 Fmoc-Thr(tBu)
35 Fmoc-Phe
36 Fmoc-Thr(tBu)
37 Fmoc-Gly
38 Fmoc-Glu(OtBu)
39 Fmoc-Dhthr
40 Boc-His(Trt)
侧链-1 Fmoc-PEG 10CH 2COOH
侧链-3 Fmoc-γGlu-OtBu
侧链-4 18烷二酸单叔丁酯
备注 Dhthr：脱羟基苏氨酸
得纯品5.6g，纯度为97.6％，总收率为10.6％。分子量为5263.8(100％M+H)。
实施例6化合物6的制备
Figure PCTCN2020115222-appb-000009
制备方法同实施例1，使用的保护氨基酸如下表:
接肽顺序n＝ 保护氨基酸
1 Fmoc-Lys(Alloc)
2 Fmoc-Ser(tBu)
3 Fmoc-Pro
4 Fmoc-Pro
5 Fmoc-Pro
6 Fmoc-Ala
7 Fmoc-Gly
8 Fmoc-Ser(tBu)
9 Fmoc-Ser(tBu)
10 Fmoc-Pro
11 Fmoc-Gly
12 Fmoc-Gly
13 Fmoc-Asn(Trt)
14 Fmoc-Lys(Boc)
15 Fmoc-Leu
16 Fmoc-Trp(Boc)
17 Fmoc-Glu(OtBu)
18 Fmoc-Ile
19 Fmoc-Phe
20 Fmoc-Leu
21 Fmoc-Arg(pbf)
22 Fmoc-Val
23 Fmoc-Ala
24 Fmoc-Glu(OtBu)
25 Fmoc-Glu(OtBu)
26 Fmoc-Glu(OtBu)
27 Fmoc-Met
28 Fmoc-Gln(Trt)
29 Fmoc-Lys(Boc)
30 Fmoc-Ser(tBu)
31 Fmoc-Leu
32 Fmoc-Asp(OtBu)
33 Fmoc-Ser(tBu)
34 Fmoc-Thr(tBu)
35 Fmoc-Phe
36 Fmoc-Thr(tBu)
37 Fmoc-Gly
38 Fmoc-Glu(OtBu)
39 Fmoc-Dhval
40 Boc-His(Trt)
侧链-1 Fmoc-PEG 10CH 2COOH
侧链-2 Fmoc-γGlu-OtBu
侧链-3 18烷二酸单叔丁酯
备注 Dhval：2,3-二脱氢缬氨酸
得纯品4.5g，纯度为97.1％，总收率为8.5％。分子量为5277.6(100％M+H)。
实施例7活性测定
1、测定方法
GLP-1R主要存在于胰岛β细胞表面，是一种G蛋白偶联的受体(GPCRs)。GLP-1R在其特异性的激动剂的刺激下，能激活细胞内腺苷酸环化酶通路，升高cAMP水平，最终导致胰岛素的生成和释放。通过待测物刺激稳定转染了GLP-1R的细胞株，使细胞胞内cAMP水平迅速升高，通过化学发光方法测定各剂量刺激细胞后的相对光单位(RLU)，进而计算出激动剂的EC50，该活性测定方法是目前国内外通用的GLP-1受体激动剂活性检测方法。
我们采用了稳定表达GLP-1R的CHO-K1细胞株，用不同浓度的激动剂刺激稳转细胞，通过测定各剂量刺激后细胞后的相对光单位，进而得到激动剂的生物活性。
2、测定结果
测定结果见下表。
化合物编号 生物活性(％)
化合物1 52.64
化合物2 18.14
化合物3 82.92
化合物4 20.25
化合物5 46.48
化合物6 14.51
实施例8初步药代特性的测定
将每个化合物分两个给药组：SD大鼠，每组雄各4只，共8只。
尾静脉静注组：剂量为1mg/kg，分别于药前(0h)、以及给药后30min、1h、2h、4h、8h、24h、48h、96h、144h大鼠眼眶静脉取血，离心分离血浆样本。
皮下给药组：剂量为1mg/kg，分别于药前(0h)、以及给药后1h、2h、3h、4h、8h、24h、48h、96h、144h大鼠眼眶静脉取血，离心分离血浆样本。
用液质联用法分别测定SD大鼠血浆样本中相应化合物的血药浓度，静脉和皮下给药后，化合物SD大鼠皮下(SC)给药半衰期见下表：
化合物 t 1/2(h)
化合物1 8.9
化合物2 8.4
化合物3 8.7
化合物4 8.9
化合物5 10.5
化合物6 10.1

Claims (7)

  1. 具有结构式Ⅰ的艾塞那肽类似物：
    AA1-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met-Glu-Glu-Glu-Ala-Val-Arg-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-AA2(R)-AA3
    结构I
    结构I中的AA1为:
    Figure PCTCN2020115222-appb-100001
    其中X 1和X 2为H，
    或为CH 3、CH(CH 3) 2、C(CH 3) 3、CH(CH 2CH 3) 2、C(CH 2CH 3) 3、CH(CH 2CH 2CH 3) 2、C(CH 2CH 2CH 3) 3、CH(CH(CH 3)) 2、C(CH(CH 3)) 3、
    或为CH 2CH 3、CH 2CH(CH 3) 2、CH 2C(CH 3) 3、CH 2CH(CH 2CH 3) 2、CH 2C(CH 2CH 3) 3、CH 2CH(CH 2CH 2CH 3) 2、CH 2C(CH 2CH 2CH 3) 3、CH 2CH(CH(CH 3)) 2、CH 2C(CH(CH 3)) 3、
    结构I中的AA2为Lys，或为Dah，或为Orn，或为Dab，或为Dap；
    结构I中的AA3为NH 2，或为OH；
    结构I中的R为HO 2C(CH 2) n1CO-(γGlu) n2-(PEG n3(CH2) n4CO) n5-
    其中：n1为10至20的整数；
    n2为1至5的整数；
    n3为1至30的整数；
    n4为1至5的整数；
    n5为1至5的整数。
  2. 根据权利要求1所述的艾塞那肽类似物，包含该类似物所成的可药用的盐、溶剂化物、螯合物或非共价复合物，基于该化合物基础上的药 物前体，或上述形式的任意混合物。
  3. 根据权利要求1和权利要求2所述的艾塞那肽类似物，用于制备治疗疾病的药物组合物。
  4. 根据权利要求3所述药物组合物，所述药物组合物在制备治疗下述至少一种疾病的药物中的用途，所述疾病包括II型糖尿病、糖耐量受损、I型糖尿病、肥胖、高血压、代谢综合征、血脂异常、认知障碍、动脉粥样硬化、心肌梗塞、冠状动脉心脏病、心血管疾病、中风、炎性肠道综合征和/或消化不良或胃溃疡、肝纤维化疾病和肺纤维化疾病。
  5. 根据权利要求4所述药物组合物，所述药物组合物在制备治疗II型糖尿病药效延迟和/或预防II型糖尿病恶化的药物中的应用。
  6. 根据权利要求6所述药物组合物，所述药物组合物在制备减少食物摄入量、减少β细胞凋亡、增加胰岛β细胞功能、增加β-细胞团和/或回复葡萄糖对β-细胞的敏感性的药物中的应用。
  7. 根据权利要求1所述艾塞那肽类似物，包含该类似物用于调节体内血糖的方法。
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113855812A (zh) * 2021-12-01 2021-12-31 上海翰森生物医药科技有限公司 聚乙二醇洛塞那肽或其药物组合物的新医药用途

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110551203B (zh) * 2019-09-25 2023-02-10 成都奥达生物科技有限公司 一种艾塞那肽类似物
CN111303245B (zh) * 2020-02-21 2023-06-27 成都奥达生物科技有限公司 一种抗合胞病毒膜融合抑制剂
CN111333714A (zh) * 2020-03-05 2020-06-26 成都奥达生物科技有限公司 一种长效glp-1化合物
CN111675752B (zh) * 2020-03-16 2023-07-07 成都奥达生物科技有限公司 一种冠状病毒膜融合抑制剂及其药物用途
CN114075275A (zh) * 2020-08-17 2022-02-22 成都奥达生物科技有限公司 一种长效胰岛素类似物
CN114478709A (zh) * 2020-11-13 2022-05-13 成都奥达生物科技有限公司 一种长效肝炎病毒进入抑制剂
CN114478694A (zh) * 2020-11-13 2022-05-13 成都奥达生物科技有限公司 一种长效mc4r激动剂
CN118684757A (zh) * 2023-03-24 2024-09-24 成都奥达生物科技有限公司 一种长效降钙素类似物

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106554404A (zh) * 2015-09-25 2017-04-05 博瑞生物医药（苏州）股份有限公司 一种艾塞那肽修饰物及其用途
WO2017178829A1 (en) * 2016-04-15 2017-10-19 Imperial Innovations Limited Peptide analogues
CN109641035A (zh) * 2016-06-29 2019-04-16 韩美药品株式会社 胰高血糖素衍生物、其缀合物、包含其的组合物、和其治疗用途
CN110551203A (zh) * 2019-09-25 2019-12-10 成都奥达生物科技有限公司 一种艾塞那肽类似物

Family Cites Families (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2003105463A (ru) * 2001-06-27 2004-11-27 Пробиодруг Аг (De) Пептидные структуры, пригодные для конкурентного модулирования катализа, осуществляемого дипептидилпептидазой iv
EP1610811A4 (en) * 2002-12-17 2008-03-26 Amylin Pharmaceuticals Inc PREVENTION AND TREATMENT OF CARDIAC ARRHYTHMIAS
KR101308912B1 (ko) * 2003-06-03 2013-09-23 노보 노르디스크 에이/에스 안정화된 약학적 펩티드 조성물
CN101365335A (zh) * 2005-12-30 2009-02-11 Ambrx公司 含有非天然氨基酸和多肽的组合物、涉及非天然氨基酸和多肽的方法以及非天然氨基酸和多肽的用途
CN101987868B (zh) * 2009-07-30 2013-09-04 江苏豪森医药集团有限公司 Glp-1类似物的衍生物或其可药用盐和用途
US8614182B2 (en) 2009-07-30 2013-12-24 Jiangsu Hansoh Pharmaceuticals Co., Ltd. GLP-1 analogues and their pharmaceutical salts and uses
JP2013530993A (ja) * 2010-07-02 2013-08-01 アンジオケム インコーポレーテッド 治療用コンジュゲートのための短く且つｄ−アミノ酸を含有するポリペプチドおよびその使用
SG11201407860PA (en) * 2012-06-14 2014-12-30 Sanofi Sa Exendin-4 peptide analogues
CN102827270A (zh) * 2012-09-13 2012-12-19 无锡和邦生物科技有限公司 一种聚乙二醇化艾塞那肽衍生物及其用途
WO2015086728A1 (en) * 2013-12-13 2015-06-18 Sanofi Exendin-4 peptide analogues as dual glp-1/gip receptor agonists
CN104945499B (zh) * 2014-03-31 2019-12-10 博瑞生物医药（苏州）股份有限公司 结构修饰的glp-1类似物及其制备方法
CN105985425B (zh) 2014-12-11 2019-12-17 浙江海昶生物医药技术有限公司 一种聚乙二醇修饰的exendin类似物及其制备方法和应用
CN110128526B (zh) * 2019-05-30 2021-07-23 江苏诺泰澳赛诺生物制药股份有限公司 长效化艾塞那肽衍生物及其盐与制备方法和用途

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106554404A (zh) * 2015-09-25 2017-04-05 博瑞生物医药（苏州）股份有限公司 一种艾塞那肽修饰物及其用途
WO2017178829A1 (en) * 2016-04-15 2017-10-19 Imperial Innovations Limited Peptide analogues
CN109641035A (zh) * 2016-06-29 2019-04-16 韩美药品株式会社 胰高血糖素衍生物、其缀合物、包含其的组合物、和其治疗用途
CN110551203A (zh) * 2019-09-25 2019-12-10 成都奥达生物科技有限公司 一种艾塞那肽类似物

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
SAMSON SUSAN L.; GARBER ALAN J.: "A Plethora of GLP-1 Agonists: Decisions About What to Use and When", CURRENT DIABETES REPORTS, CURRENT SCIENCE, PHILADELPHIA, VA, US, vol. 16, no. 12, 20 October 2016 (2016-10-20), US, pages 1 - 13, XP036115283, ISSN: 1534-4827, DOI: 10.1007/s11892-016-0823-6 *
See also references of EP4036108A4 *
YI FAN, DU QUAN: "GLP-1 biology and GLP-1 based antidiabetic therapy", JOURNAL OF CHINESE PHARMACEUTICAL SCIENCES, SCHOOL OF PHARMACEUTICAL SCIENCES, BEIJING MEDICAL UNIVERSITY, BEIJING , CN, vol. 22, no. 1, 15 January 2013 (2013-01-15), Beijing , CN, XP055793851, ISSN: 1003-1057, DOI: 10.5246/jcps.2013.01.001 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113855812A (zh) * 2021-12-01 2021-12-31 上海翰森生物医药科技有限公司 聚乙二醇洛塞那肽或其药物组合物的新医药用途

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