WO2011063549A1 - Analogues d'exendine 4 à action prolongée - Google Patents

Analogues d'exendine 4 à action prolongée Download PDF

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Publication number
WO2011063549A1
WO2011063549A1 PCT/CN2009/001321 CN2009001321W WO2011063549A1 WO 2011063549 A1 WO2011063549 A1 WO 2011063549A1 CN 2009001321 W CN2009001321 W CN 2009001321W WO 2011063549 A1 WO2011063549 A1 WO 2011063549A1
Authority
WO
WIPO (PCT)
Prior art keywords
glu
leu
gly
exendin
diabetes
Prior art date
Application number
PCT/CN2009/001321
Other languages
English (en)
Chinese (zh)
Inventor
吴晓琰
龚铁军
孙玉琨
Original Assignee
Wu Xiaoyan
Gong Tiejun
Sun Yukun
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Wu Xiaoyan, Gong Tiejun, Sun Yukun filed Critical Wu Xiaoyan
Priority to PCT/CN2009/001321 priority Critical patent/WO2011063549A1/fr
Priority to CN200980162062.XA priority patent/CN102712690B/zh
Publication of WO2011063549A1 publication Critical patent/WO2011063549A1/fr

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Classifications

    • 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
    • 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
    • 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

Definitions

  • the present invention relates to a polypeptide of an Exendin 4 analog which has a hypoglycemic effect and is useful for the treatment of type 2 diabetes.
  • the invention also provides methods of producing these polypeptides by chemical synthesis and recombinant DNA production processes.
  • type 1 diabetes insulin-dependent diabetes
  • type 2 diabetes non-insulin-dependent diabetes
  • Type 2 diabetic patients exhibit many characteristics, such as decreased ⁇ -cells, insufficient insulin secretion after meals, delayed secretion of insulin, increased fasting and postprandial blood glucose, elevated glycosylated hemoglobin HbAlc, and insulin resistance.
  • Exendin 4 and Exendin 3 are two peptides secreted by the saliva of Gila monster, Heloderme Suspectum, which consists of 39 amino acid residues, chemically synthesized as an anti-type 2 diabetes drug. Approved by the US FDA in 2005, the trade name is Byetta, and it was listed in China in 2009.
  • Exendin 4 can promote the synthesis of proinsulin, promote insulin secretion, reduce fasting, postprandial blood glucose, and no longer continue to function when blood sugar is normal, so it is not easy to produce hypoglycemia, coma, shock, and high safety. It can reduce HbAl c , increase the number and volume of pancreatic ⁇ cells, increase the sensitivity of insulin receptors in type 2 diabetes patients, and inhibit the secretion of glucagon. Inject twice a day, 5-10ug each time.
  • Exendin 4 is derived from the giant lizard, and GLP-1 is a human endogenous substance.
  • Exendin 4 is a chemically synthesized product that is very expensive.
  • the object of the present invention is to address the current market demand for Exendin 4, which is very expensive, and the present specification provides a 32 amino acid polypeptide derivative of Exendin 4. Its peptide chain length is similar to that of human GLP-1, reducing its immunogenicity.
  • the structure contains only one Lys20 residue, which allows the fatty acid acylation to specifically bind to Lys20 ⁇ (epsi lon ) - ⁇ 2 to form a fatty acid acylated derivative.
  • the present invention utilizes genetic engineering technology to mass-produce the polypeptide, which can reduce the cost and provide a new therapeutic drug for a large number of type 2 diabetic patients.
  • One aspect of the present invention provides a structural change, and still has hypoglycemic activity Exendin 4 Analog,
  • the Exendin 4 analog of the present invention has the following structural formula:
  • X2 Met, Leu or He
  • X3 Arg, OH or NH2.
  • XI is Tyr or Phe
  • X2 is Met or Leu
  • X3 is Arg
  • compositions comprising the above Exendin 4 analogue, the composition comprising one or more compounds having the above formula and a pharmaceutically acceptable diluent or excipient, preferably
  • the composition is in unit dosage form, such as tablets, pills, capsules (including sustained release or delayed release forms), powders, granules, elixirs, elixirs, syrups and emulsions, sterile injectable solutions or suspensions, aerosols Or liquid sprays, drops, injections, automatic injection devices or suppositories.
  • the above active pharmaceutical ingredient may be combined with an oral non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like.
  • an oral non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like.
  • the present invention provides the use of the above compound and a pharmaceutical combination for the preparation of a therapeutic disease, preferably the disease is type 2 diabetes.
  • a further aspect of the invention provides a genetic engineering method for the preparation of the above Exendi n 4 analogs and derivatives.
  • the above compound in which X3 is Arg may be synthesized by a genetic engineering method or may be synthesized by a chemical method, but is preferably synthesized by a genetic engineering method.
  • the above compounds in which X3 is 0H or NH2 can also be synthesized by a chemical synthesis method using a polypeptide synthesizer.
  • the invention changes the chemical structure of Exend in 4, shortens the length of the peptide chain from the C-terminus and changes the amino acid structure, and maintains its hypoglycemic activity, so that the peptide chain length of Exendi n 4 is similar to that of human GLP-1, and the immunogenicity is lowered.
  • the peptide chain contains only one Lys20, which facilitates the site-direct acylation of fatty acids to form long-acting derivatives.
  • the need to adapt to the production of genetic engineering methods is suitable for mass production of genetic engineering, reducing costs, adapting to the needs of patients with type 2 diabetes, and providing a new therapeutic drug for patients with type 2 diabetes.
  • Figure 1 is a tandem map of Exendi n 4 analog gene fragments
  • Figure 2 is a flow chart of the production of Exendi n 4 analog by genetic engineering
  • Figure 3 is a measure of the hypoglycemic effect of EW1 and Exend i n4.
  • Embodiment 1 is a diagrammatic representation of Embodiment 1:
  • the EW1 gene was synthesized by the conventional PCR method, and then digested with EcoR I /Sal I and cloned into the Lac promoter-containing plasmid.
  • the DNA sequence was identical to the design.
  • 487f 5' ccaatgAATTCCAGATCTAACGGCCGTCACGGCGAAG 3' 487r : 5' aattGTCGACTAGGATCCACGCGGGCCGCCCTGAACCAGCCAT 3'
  • Ampicillin (Ampici ll in ) to a final concentration of 50 ⁇ g/ml, 37 ° C, shaken overnight
  • the culture medium is composed of M9 medium, with glucose concentration of 1%, ampicillin (Ampici ll in ) concentration of 50 g/ml, aeration of 20L/ m in, dissolved oxygen. Maintained at more than 20%, the antifoaming agent is domestically produced, the pH is maintained at 7 ⁇ 8, and it is adjusted with ammonia water. When the concentration of the bacteria reached the neutral line of the log curve, IPTG was added, and the concentration was 0.5 mM. The fermentation was continued for 4 to 6 hours, and the cells were collected by centrifugation to a wet weight of 30 to 50 g/L.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Diabetes (AREA)
  • General Health & Medical Sciences (AREA)
  • Endocrinology (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Vascular Medicine (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Obesity (AREA)
  • Emergency Medicine (AREA)
  • Immunology (AREA)
  • Hematology (AREA)
  • Epidemiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Toxicology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention concerne un peptide analogue d'exendine 4. Par comparaison à l'exendine 4 de type sauvage, le peptide de la présente invention est plus court de 7 résidus d'acides aminés à son extrémité C-terminale. La longueur de la chaîne peptidique est similaire à GLP-1 humain, et il n'y a qu'un résidu d'acide aminé Lys20 dans la structure du peptide, de telle sorte que l'acylation d'acide gras se produit spécifiquement au ε-NH2 de la Lys20 pour former un dérivé acylé d'acide gras. L'invention concerne également des compositions pharmaceutiques comprenant un tel peptide et des procédés pour produire le peptide par synthèse chimique ou par des technologies d'ADN recombinant.
PCT/CN2009/001321 2009-11-26 2009-11-26 Analogues d'exendine 4 à action prolongée WO2011063549A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
PCT/CN2009/001321 WO2011063549A1 (fr) 2009-11-26 2009-11-26 Analogues d'exendine 4 à action prolongée
CN200980162062.XA CN102712690B (zh) 2009-11-26 2009-11-26 长效Exendin 4的类似物

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2009/001321 WO2011063549A1 (fr) 2009-11-26 2009-11-26 Analogues d'exendine 4 à action prolongée

Publications (1)

Publication Number Publication Date
WO2011063549A1 true WO2011063549A1 (fr) 2011-06-03

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ID=44065807

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2009/001321 WO2011063549A1 (fr) 2009-11-26 2009-11-26 Analogues d'exendine 4 à action prolongée

Country Status (2)

Country Link
CN (1) CN102712690B (fr)
WO (1) WO2011063549A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2017510636A (ja) * 2014-03-21 2017-04-13 エニジェン カンパニー．，リミテッド． 新規なエキセナチド類似体及びその用途

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103193881A (zh) * 2013-04-22 2013-07-10 中国药科大学 一种可口服给药的降糖多肽衍生物及其用途

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1724563A (zh) * 2005-06-29 2006-01-25 常州制药厂有限公司 Exendin4多肽片段
CN1746185A (zh) * 2004-09-06 2006-03-15 上海华谊生物技术有限公司 Exendin 4的类似物
US7329646B2 (en) * 2001-05-10 2008-02-12 Shanghai Huayi Bio-Lab Co., Ltd. Derivatives of the insulinotropic peptide exendin-4 and methods of production thereof
WO2008023050A1 (fr) * 2006-08-25 2008-02-28 Novo Nordisk A/S Composés d'exendine-4 acylée

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7329646B2 (en) * 2001-05-10 2008-02-12 Shanghai Huayi Bio-Lab Co., Ltd. Derivatives of the insulinotropic peptide exendin-4 and methods of production thereof
CN1746185A (zh) * 2004-09-06 2006-03-15 上海华谊生物技术有限公司 Exendin 4的类似物
CN1724563A (zh) * 2005-06-29 2006-01-25 常州制药厂有限公司 Exendin4多肽片段
WO2008023050A1 (fr) * 2006-08-25 2008-02-28 Novo Nordisk A/S Composés d'exendine-4 acylée

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
SON, S. ET AL.: "Preparation and Structural, Biochemical, and Pharmaceutical Characterizations of Bile Acid-Modified Long-Acting Exendin-4 Derivatives.", J. MED. CHEM., vol. 52, no. 21, 12 November 2009 (2009-11-12), pages 6889 - 6896, XP008157488, DOI: doi:10.1021/jm901153x *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2017510636A (ja) * 2014-03-21 2017-04-13 エニジェン カンパニー．，リミテッド． 新規なエキセナチド類似体及びその用途
CN106661096A (zh) * 2014-03-21 2017-05-10 安尼根有限公司 新的艾塞那肽类似物及其用途
EP3121195A4 (fr) * 2014-03-21 2017-10-11 Anygen Co., Ltd. Nouvel analogue de l'exénatide et son utilisation
US11103557B2 (en) 2014-03-21 2021-08-31 Anygen Co., Ltd. Exenatide analogue and use thereof

Also Published As

Publication number Publication date
CN102712690B (zh) 2016-04-13
CN102712690A (zh) 2012-10-03

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