CN111333714A - Long-acting GLP-1 compound - Google Patents
Long-acting GLP-1 compound Download PDFInfo
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- CN111333714A CN111333714A CN202010147530.4A CN202010147530A CN111333714A CN 111333714 A CN111333714 A CN 111333714A CN 202010147530 A CN202010147530 A CN 202010147530A CN 111333714 A CN111333714 A CN 111333714A
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- C07—ORGANIC CHEMISTRY
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- 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
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- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Abstract
The invention relates to the field of medicine synthesis, and discloses a long-acting GLP-1 compound. The long-acting GLP-1 compound of the invention is used for preparing a pharmaceutical composition for treating diseases, and the application of the pharmaceutical composition in preparing a medicament for treating at least one disease of type II diabetes, impaired glucose tolerance, type I diabetes, obesity, hypertension, metabolic syndrome, dyslipidemia, cognitive disorder, atherosclerosis, myocardial infarction, coronary heart disease, cardiovascular disease, stroke, inflammatory bowel syndrome and/or dyspepsia or gastric ulcer, hepatic fibrosis disease and pulmonary fibrosis disease.
Description
Technical Field
The invention relates to a long-acting GLP-1 compound and application thereof, wherein the analogue is a glucagon-like peptide-1 (GLP-1) analogue.
Background
Diabetes has become the third non-infectious disease after cardiovascular and cerebrovascular disease and tumor, and the World Health Organization (WHO) predicts: in 2030, the number of diabetic patients worldwide will exceed 3.6 hundred million, of which more than 90% are type II diabetes. GLP-1 is a secretive from intestinal tract L cell, it has effects of promoting insulin secretion, inhibiting release of glucagon, stimulating B cell proliferation of pancreatic island, inducing B cell regeneration of pancreatic island, preventing B cell apoptosis of pancreatic island, improving insulin sensitivity and increasing glucose utilization, etc., and plays an important role in the occurrence and development of type II diabetes. A patient with type II diabetes is impaired in an intestinal insulinotropic effect, and the increase amplitude of the GLP-1 concentration after meal intake is reduced compared with that of a normal person, but the effects of promoting insulin secretion and reducing blood sugar are not obviously impaired, so that the GLP-1 can be used as an important target point for treating type II diabetes, and meanwhile, the GLP-1 has glucose concentration dependence, and the blood sugar reduction characteristic is the basis and guarantee of clinical application safety, so that the worry that serious hypoglycemia of the patient can be caused by the existing diabetes treatment medicines and schemes is avoided, and the method has a wide application prospect in the field of diabetes treatment.
However, the clinical application of GLP-1 faces huge problems, the GLP-1 generated by a human body is very unstable and is very easy to be degraded by dipeptidyl peptidase IV (DPP-IV) in vivo, and the plasma half-life period of the GLP-1 is short, so that the clinical application of the GLP-1 is limited. Also, many type II diabetics are reluctant to administer daily injections, so there is great promise in developing safe and effective GLP-1 analogues that can be administered once a week.
Disclosure of Invention
The invention provides a long-acting GLP-1 compound and application thereof, wherein the compound is a glucagon-like peptide-1 (GLP-1) analogue.
To achieve the above object, the present invention provides, in a first aspect, a compound of structure I, a pharmaceutically acceptable salt, solvate, chelate or non-covalent complex thereof, a prodrug based on the compound, or any mixture thereof.
His-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-AA2-AA3(R)-AA4
Structure I
When AA2 in structure I is Ser, AA1 is Aib, or Acpr, or Acp, or Acpe, or Ach;
AA1 is Aib, or Acpr, or Acpe, or Ach, when AA2 is absent in structure I; or Dhthr, or DhVal;
AA3 in structure I is Lys, or Dah, or Orn, or Dab, or Dap;
AA4 in Structure I is NH2Or is OH;
r in structure I is HO2C(CH2)n1CO-(γGlu)n2-(PEGn3(CH2)n4CO)n5-
Wherein: 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 invention also provides pharmaceutical compositions comprising a compound according to the invention and the use of a pharmaceutical composition comprising a compound of the invention for the preparation of a medicament for the treatment of a disease.
Preferably, the use of the pharmaceutical composition in the manufacture of a medicament for the treatment of at least one of type II diabetes, impaired glucose tolerance, type I diabetes, obesity, hypertension, metabolic syndrome, dyslipidemia, cognitive disorders, atherosclerosis, myocardial infarction, coronary heart disease, cardiovascular disease, stroke, inflammatory bowel syndrome and/or dyspepsia or gastric ulcer, liver fibrosis diseases and pulmonary fibrosis diseases.
Preferably, the pharmaceutical composition is applied to the preparation of medicines for treating delayed drug effect and/or preventing worsening of type II diabetes.
Preferably, the use of the pharmaceutical composition for the manufacture of a medicament for reducing food intake, reducing β apoptosis, increasing pancreatic islet β cell function, increasing β -cell mass, and/or restoring glucose sensitivity to β -cells.
The invention still further provides methods of administering the compounds to a subject for the modulation of blood glucose in vivo.
Further details of the invention are set forth below, or some may be appreciated in embodiments of the invention.
Unless otherwise indicated, the amounts of the various ingredients, reaction conditions, and the like used herein are to be construed in any case to mean "about". Accordingly, unless expressly stated otherwise, all numerical parameters set forth in the following specification and attached claims are approximations that may vary depending upon the standard deviation found in the respective experimental conditions.
Herein, when a chemical structural formula and a chemical name of a compound are ambiguous or ambiguous, the compound is exactly defined by the chemical structural formula. The compounds described herein may contain one or more chiral centers, and/or double bonds and the like, and stereoisomers, including isomers of double bonds (e.g., geometric isomers), optical enantiomers, or diastereomers, may also be present. Accordingly, any chemical structure within the scope of the description, whether partial or complete, including similar structures as described above, includes all possible enantiomers and diastereomers of the compound, including any stereoisomer alone (e.g., pure geometric isomers, pure enantiomers, or pure diastereomers), as well as any mixture of such stereoisomers. Mixtures of these racemates and stereoisomers may also be further resolved into the enantiomers or stereoisomers of their constituent members by those skilled in the art using non-stop separation techniques or methods of chiral molecular synthesis.
The compounds of formula I include, but are not limited to, optical isomers, racemates and/or other mixtures of these compounds. In the above case, a single enantiomer or diastereomer, such as an optical isomer, can be obtained by asymmetric synthesis or racemate resolution. Resolution of the racemates can be accomplished by various methods, such as conventional recrystallization from resolution-assisting reagents, or by chromatographic methods. In addition, the compounds of formula I also include cis and/or trans isomers with double bonds.
The compounds of the present invention include, but are not limited to, the compounds of formula I and all of their pharmaceutically acceptable different forms. The pharmaceutically acceptable different forms of these compounds include various pharmaceutically acceptable salts, solvates, complexes, chelates, non-covalent complexes, prodrugs based on the above and any mixtures of these forms.
The compound shown in the structure I provided by the invention has stable property, is not easily degraded by dipeptidyl peptidase IV (DPP-IV) in vivo, is a long-acting GLP-I compound, and has a remarkable hypoglycemic effect.
Detailed Description
The invention discloses a long-acting GLP-1 compound and application thereof, and a person skilled in the art can appropriately improve related parameters by referring to the content. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the process of the present invention has been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications of the compounds and processes described herein, as well as other changes and combinations of the foregoing, may be made to implement and use the techniques of the present invention without departing from the spirit and scope of the invention.
The Chinese names corresponding to the English abbreviations related in the invention are shown in the following table:
english abbreviation | Name of Chinese | English abbreviation | Name of Chinese |
Fmoc | 9-fluorenylmethoxycarbonyl group | OtBu | Tert-butoxy radical |
tBu | Tert-butyl radical | Boc | Boc-acyl |
Trt | Trityl radical | Pbf | (2, 3-dihydro-2, 2,4,6, 7-pentamethylbenzofuran-5-yl) sulfonyl group |
Ala | Alanine | Leu | Leucine |
Arg | Arginine | Lys | Lysine |
Asn | Asparagine | Met | Methionine |
Asp | Aspartic acid | Phe | Phenylalanine |
Cys | Cysteine | Pro | Proline |
Gln | Glutamine | Ser | Serine |
Glu | Glutamic acid | Thr | Threonine |
Gly | Glycine | Trp | Tryptophan |
His | Histidine | Tyr | Tyrosine |
Ile | Isoleucine | Val | Valine |
Dap | 2, 3-diaminopropionic acid | Dab | 2, 4-diaminobutyric acid |
Orn | Ornithine | Dah | 2, 7-Diaminoheptanoic acid |
Aib | 2-Aminoisobutyric acid | Acpr | 1-aminocyclopropanecarboxylic acid |
Acp | 1-aminocyclobutanecarboxylic acid | Acpe | 1-aminocyclopentanoic acid |
Ach | 1-aminocyclohexanecarboxylic acid | Dhthr | Dehydroxythreonine |
Dhval | 2, 3-didehydro valine |
EXAMPLE 1 preparation of Compound 1
His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met-Glu-Glu-Ala-Val-Arg-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-Gly-Ala-Pro-Pro-Pro-Ser-Lys (AEEA-AEEA-gamma Glu-octadecanedioic acid) -NH2
The preparation method comprises the following steps: preparing peptide resin by adopting a solid-phase polypeptide synthesis method, carrying out acidolysis on the peptide resin to obtain a crude product, and finally purifying the crude product to obtain a pure product; the step of preparing the peptide resin by the solid-phase polypeptide synthesis method is to sequentially insert corresponding protective amino acids or fragments in the following sequences on a carrier resin by the solid-phase coupling synthesis method to prepare the peptide resin:
in the preparation method, the dosage of the Fmoc-protected amino acid or the protected amino acid fragment is 1.2-6 times of the total mole number of the charged resin; preferably 2.5 to 3.5 times.
In the preparation method, the substitution value of the carrier resin is 0.2-1.0 mmol/g resin, and the preferable substitution value is 0.3-0.5 mmol/g resin.
In a preferred embodiment of the present invention, the solid-phase coupling synthesis method comprises: and (3) after the Fmoc protecting group of the protected amino acid-resin obtained in the previous step is removed, carrying out coupling reaction with the next protected amino acid. The deprotection time for removing Fmoc protection is 10-60 minutes, and preferably 15-25 minutes. The coupling reaction time is 60-300 minutes, and preferably 100-140 minutes.
The coupling reaction needs to add a condensation reagent, and the condensation reagent is selected from one of DIC (N, N-diisopropyl carbodiimide), N, N-dicyclohexylcarbodiimide, benzotriazole-1-yl-oxy tripyrrolidinophosphonium hexafluorophosphate, 2- (7-aza-1H-benzotriazole-1-yl) -1,1,3, 3-tetramethylurea hexafluorophosphate, benzotriazole-N, N, N ', N' -tetramethylurea hexafluorophosphate or O-benzotriazole-N, N, N ', N' -tetramethylurea tetrafluoroborate; n, N-diisopropylcarbodiimide is preferred. The molar consumption of the condensation reagent is 1.2-6 times of the total molar number of amino groups in the amino resin, and preferably 2.5-3.5 times.
The coupling reaction needs to add an activating reagent, wherein the activating reagent is selected from 1-hydroxybenzotriazole or N-hydroxy-7-azabenzotriazole, and 1-hydroxybenzotriazole is preferred. The amount of the activating agent is 1.2 to 6 times, preferably 2.5 to 3.5 times of the total mole number of the amino groups in the amino resin.
As a preferable scheme of the invention, the reagent for removing Fmoc protection is PIP/DMF (piperidine/N, N-dimethylformamide) mixed solution, and the piperidine content in the mixed solution is 10-30% (V). The dosage of the Fmoc protection removing reagent is 5-15 mL per gram of amino resin, and preferably 8-12 mL per gram of amino resin.
Preferably, the peptide resin is subjected to acidolysis while removing the resin and side chain protecting groups to obtain a crude product:
more preferably, the acidolysis agent used in the acidolysis 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 as follows: 80-95% of TFA, 1-10% of EDT and the balance of water.
More preferably, the volume ratio of the mixed solvent is: 89-91% of TFA, 4-6% of EDT and the balance of water. Optimally, the volume ratio of the mixed solvent is as follows: TFA 90%, EDT 5%, balance water.
The dosage of the acidolysis agent is 4-15 mL per gram of the peptide resin; preferably, 7-10 mL of acidolysis agent is required per gram of peptide resin.
The time for cracking by using the acidolysis agent is 1-6 hours, preferably 3-4 hours at room temperature.
Further, the crude product is purified by high performance liquid chromatography and freeze-dried to obtain a pure product.
1. Synthesis of peptide resins
Rink Amide BHHA resin is used as carrier resin, and is coupled with protected amino acid shown in the following table in sequence through Fmoc protection removal and coupling reaction to prepare peptide resin. The protected amino acids used in this example correspond to the protected amino acids shown below:
(1) 1 st protected amino acid inserted into main chain
Dissolving 0.03mol of the 1 st protected amino acid and 0.03mol of HOBt in a proper amount of DMF; and adding 0.03mol DIC slowly into the protected amino acid DMF solution under stirring, and reacting for 30 minutes under stirring at room temperature to obtain an activated protected amino acid solution for later use.
0.01mol of Rink amide MBHA resin (substitution value about 0.4mmol/g) is taken, deprotected by 20% PIP/DMF solution for 25 minutes, washed and filtered to obtain Fmoc-removed resin.
And adding the activated 1 st protected amino acid solution into the Fmoc-removed resin, performing coupling reaction for 60-300 minutes, and filtering and washing to obtain the resin containing 1 protected amino acid.
(2) 2-40 protected amino acids connected to main chain
And sequentially inoculating the corresponding 2 nd to 40 th protected amino acids by the same method for inoculating the 1 st protected amino acid of the main chain to obtain the resin containing 40 amino acids of the main chain.
(3) Side chain insertion of the 1 st protected amino acid
Dissolving 0.03mol of the 1 st protected amino acid of the side chain and 0.03mol of HOBt in a proper amount of DMF; and adding 0.03mol DIC slowly into the protected amino acid DMF solution under stirring, and reacting for 30 minutes under stirring at room temperature to obtain an activated protected amino acid solution.
Taking 2.5mmol of tetratriphenylphosphine palladium and 25mmol of phenylsilane, dissolving with a proper amount of dichloromethane, deprotecting for 4 hours, filtering and washing to obtain a resin without Alloc for later use.
Adding the activated side chain 1 st protected amino acid solution into the Alloc-removed resin, performing coupling reaction for 60-300 minutes, filtering and washing to obtain the side chain 1 st protected amino acid-containing resin.
(4) 2-4 protective amino acids of grafted side chain
And sequentially inoculating 2 nd to 4 th protected amino acids and single protected fatty acids corresponding to side chains by adopting the same method for inoculating the 1 st protected amino acid to the main chain to obtain the peptide resin.
2. Preparation of crude product
Adding a cleavage reagent (10 mL of cleavage reagent/g of resin) with the volume ratio of TFA, water and EDT (95: 5) into the peptide resin, uniformly stirring, stirring at room temperature for reaction for 3 hours, filtering a reaction mixture by using a sand core funnel, collecting filtrate, washing the resin with a small amount of TFA for 3 times, combining the filtrates, concentrating under reduced pressure, adding anhydrous ether for precipitation, washing the precipitate with anhydrous ether for 3 times, and drying to obtain white-like powder, namely a crude product.
3. Preparation of the pure product
Mixing the crude product with water, stirring, adjusting pH to 8.0 with ammonia water to dissolve completely, filtering the solution with 0.45 μm mixed microporous membrane, and purifying;
purifying by high performance liquid chromatography, wherein the chromatographic packing for purification is 10 μm reversed phase C18, the mobile phase system is 0.1% TFA/water solution-0.1% TFA/acetonitrile solution, the flow rate of a 30mm by 250mm chromatographic column is 20mL/min, eluting by a gradient system, circularly sampling for purification, sampling the crude product solution in the chromatographic column, starting the mobile phase for elution, collecting the main peak, and evaporating acetonitrile to obtain a purified intermediate concentrated solution;
filtering the purified intermediate concentrated solution with 0.45 μm filter membrane for use, and performing salt exchange by high performance liquid chromatography with 1% acetic acid/water solution-acetonitrile as mobile phase system, 10 μm reversed phase C18 as purification chromatographic filler, and 20mL/min of 30 mm/250 mm chromatographic column flow rate (corresponding flow rate can be adjusted according to chromatographic columns of different specifications); adopting gradient elution and circulation sample loading method, loading sample in chromatographic column, starting mobile phase elution, collecting atlas, observing change of absorbance, collecting main peak of salt exchange and analyzing liquid phase to detect purity, combining main peak solutions of salt exchange, concentrating under reduced pressure to obtain pure acetic acid water solution, and freeze drying to obtain pure acetic acid 5.7g, purity 98.2% and total yield 11.3%. The molecular weight was 5056.2 (100% M + H).
EXAMPLE 2 preparation of Compound 2
His-Acpr-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met-Glu-Glu-Ala-Val-Arg-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-Gly-Ala-Pro-Pro-Pro-Ser-Lys (AEEA-AEEA-Gamma Glu-octadecanedioic acid) -NH2
The procedure is as in example 1, using the protected amino acids as in the following table:
6.0g of pure product is obtained, the purity is 97.1 percent, and the total yield is 11.8 percent. The molecular weight was 5070.7 (100% M + H).
EXAMPLE 3 preparation of Compound 3
His-Acp-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met-Glu-Glu-Ala-Val-Arg-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-Gly-Ala-Pro-Pro-Pro-Ser-Lys (AEEA-AEEA-gamma Glu-octadecanedioic acid) -NH2
The procedure is as in example 1, using the protected amino acids as in the following table:
7.2g of pure product is obtained, the purity is 97.6 percent, and the total yield is 14.2 percent. The molecular weight was 5084.6 (100% M + H).
EXAMPLE 4 preparation of Compound 4
His-Acpe-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-Lys (AEEA-AEEA-gamma Glu-octadecanedioic acid) -NH2
The procedure is as in example 1, using the protected amino acids as in the following table:
6.3g of pure product is obtained, the purity is 98.9 percent, and the total yield is 12.4 percent. The molecular weight was 5098.8 (100% M + H).
EXAMPLE 5 preparation of Compound 5
His-Ach-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met-Glu-Glu-Ala-Val-Arg-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-Gly-Ala-Pro-Pro-Pro-Ser-Lys (AEEA-AEEA-gamma Glu-octadecanedioic acid) -NH2
The procedure is as in example 1, using the protected amino acids as in the following table:
6.9g of pure product is obtained, the purity is 97.1 percent, and the total yield is 13.5 percent. The molecular weight was 5112.8 (100% M + H).
Example 6 Activity assay
1. Measurement method
GLP-1R mainly exists on the cell surface of pancreatic islet β and is a G Protein Coupled Receptor (GPCRs). GLP-1R can activate intracellular adenylate cyclase pathway under the stimulation of specific agonist, increase cAMP level, and finally cause the generation and release of insulin.A cell strain stably transfected with GLP-1R is stimulated by an analyte to ensure that intracellular cAMP level is rapidly increased, Relative Light Unit (RLU) after each dose of stimulated cells is determined by a chemiluminescence method, and then EC50 of the agonist is calculated.
A CHO-K1 cell strain which stably expresses GLP-1R is adopted, agonist with different concentrations is used for stimulating stable cells, relative light units after the cells are stimulated by various doses are measured, exenatide is used as a reference substance, and then the relative biological activity of the agonist is obtained.
2. Measurement results
The results of the measurements are shown in the following table.
Compound numbering | Biological Activity (%) |
Compound 1 | 65.17 |
Compound 2 | 57.39 |
Compound 3 | 52.65 |
Compound 4 | 45.48 |
Compound 5 | 30.52 |
EXAMPLE 7 determination of Primary pharmacokinetic Properties
Each compound was divided into two dosing groups: SD rats, 4 males per group, 8 in total.
Tail vein intravenous injection group: the dose is 1mg/kg, rat orbital veins are respectively bled before (0h) and 30min, 1h, 2h, 4h, 8h, 24h, 48h, 96h and 144h after administration, and plasma samples are centrifugally separated.
Subcutaneous administration group: the dose is 1mg/kg, rat orbital veins are respectively bled before (0h) and 1h, 2h, 3h, 4h, 8h, 24h, 48h, 96h and 144h after administration, and plasma samples are separated by centrifugation.
Plasma concentrations of the corresponding compounds in plasma samples of SD rats were measured by the liquid chromatography-mass spectrometry method, and the half-lives of the compounds after intravenous and subcutaneous administration in SD rats under Subcutaneous (SC) administration are shown in the following table:
compound (I) | t1/2(h) |
Compound 1 | 7.2 |
Compound 2 | 8.5 |
Compound 3 | 9.1 |
Compound 4 | 9.8 |
Compound 5 | 11.2 |
Claims (7)
1. A long-acting GLP-1 compound having the structural formula i:
His-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-AA2-AA3(R)-AA4
structure I
When AA2 in structure I is Ser, AA1 is Aib, or Acpr, or Acp, or Acpe, or Ach;
AA1 is Aib, or Acpr, or Acpe, or Ach, when AA2 is absent in structure I; or Dhthr, or DhVal;
AA3 in structure I is Lys, or Dah, or Orn, or Dab, or Dap;
AA4 in Structure I is NH2Or is OH;
r in structure I is HO2C(CH2)n1CO-(γGlu)n2-(PEGn3(CH2)n4CO)n5-
Wherein: 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.
2. The long-acting GLP-1 compound according to claim 1, comprising a pharmaceutically acceptable salt, solvate, chelate or non-covalent complex of said analogue, a prodrug based on said compound, or a mixture of any of the above forms.
3. The long-acting GLP-1 compound according to claim 1 and claim 2 for the preparation of a pharmaceutical composition for the treatment of a disease.
4. The pharmaceutical composition according to claim 3, for use in the manufacture of a medicament for the treatment of at least one of type II diabetes, impaired glucose tolerance, type I diabetes, obesity, hypertension, metabolic syndrome, dyslipidemia, cognitive disorders, atherosclerosis, myocardial infarction, coronary heart disease, cardiovascular disease, stroke, inflammatory bowel syndrome and/or dyspepsia or gastric ulcer, liver fibrosis diseases and pulmonary fibrosis diseases.
5. The pharmaceutical composition according to claim 4, for use in the preparation of a medicament for the treatment of delayed efficacy and/or prevention of exacerbation of type II diabetes.
6. The pharmaceutical composition of claim 6, for use in the preparation of a medicament for reducing food intake, reducing β apoptosis, increasing pancreatic islet β cell function, increasing β -cell mass, and/or restoring glucose sensitivity to β -cells.
7. The long-acting GLP-1 compound of claim 1, comprising the analog for use in a method of modulating blood glucose in vivo.
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