Long-acting dual agonist compound

Abstract

The present invention relates to the field of pharmaceutical combination components and discloses a GLP-1 / Gcg dual agonist compound. The GLP-1 / Gcg dual agonist compound described above is used for manufacturing a pharmaceutical composition for treating diseases. The pharmaceutical composition is used for manufacturing a medicament for treating at least one of diseases including type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, metabolic syndrome, dyslipidemia, cognitive impairment, atherosclerosis, myocardial infarction, coronary heart disease, cardiovascular disease, stroke, inflammatory bowel syndrome and / or indigestion or gastric ulcer, liver fibrosis and pulmonary fibrosis.

Description

Technical Field

[0001] [Cross-reference] This application claims priority based on a Chinese patent application filed with the China National Intellectual Property Administration on August 10, 2022, with an application number of 202210957875.5 and an invention title of "Long-acting dual agonist compound", and the entire content thereof is incorporated herein by reference.

[0002] The present invention relates to the field of pharmaceutical compositions, and in particular, to a long-acting dual agonist compound which is a dual agonist compound of a type of glucagon (Gcg) receptor and glucagon-like peptide-1 (GLP-1) receptor.

Background Art

[0003] GLP-1 is a 37-amino acid peptide that stimulates insulin secretion, protects pancreatic β-cells, inhibits glucagon secretion, gastric emptying and food intake, thereby leading to weight loss. GLP-1 is called an incretin. Incretin receptor signaling plays a physiologically important role in glucose homeostasis. In normal physiology, GLP-1 is secreted from the gastrointestinal tract after a meal. These incretins enhance physiological responses to food, including satiety, insulin secretion, and nutrient processing.

[0004] The most common side effect of GLP-1 analogs is that their administration cannot achieve sufficient and effective blood glucose control and weight loss, while GIP alone has a very moderate blood glucose lowering ability for type 2 diabetes patients. Since natural GLP-1 can be rapidly inactivated by the generally present protease DPP IV, it is only used for short-term metabolic management.

[0005] Gcg, also known as glucagon or anti-insulin or insulin B, is a hormone secreted from pancreatic α-cells of the vertebrate pancreas together with insulin, antagonizes insulin, and has the function of increasing blood glucose levels.

[0006] In the latest research, it has been shown that GLP-1 / Gcg receptor dual agonists not only have a better blood glucose control effect, but also have an obvious effect on the treatment of weight loss and non-alcoholic fatty liver disease.

[0007] Currently, GLP-1 / Gcg receptor dual agonists are not yet commercially available. OXM3, as a GLP-1 / Gcg receptor dual agonist compound in clinical research, has a half-life of 120 hours and is administered at a dosing frequency of once a week.

Summary of the Invention

Problems to be Solved by the Invention

[0008] In view of the above circumstances, the present invention provides a long-acting dual agonist compound, which is a dual agonist compound of a kind of glucagon (Gcg) receptor and glucagon-like peptide-1 (GLP-1) receptor.

Means for Solving the Problems

[0009] In order to achieve the above object, the present invention provides the following technical methods. According to the present invention, there is provided a compound comprising the following (I) or (II) or (III) or (IV) and / or (V) and / or (VI): (I) The amino acid sequence represented by Formula I; His-AA1-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Lys-Lys-Ala-Lys(R)-Glu-Phe-Val-Glu-Trp-Leu-Leu-AA2-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-AA3 [Formula I] [In Formula I, AA1 is selected from the group consisting of Aib, Acpr, Acp, Acpe and Ach, AA2 is Glu or Ser, AA3 is NH2 or OH, R is HO2C(CH2) n1 CO-(AA4) n2 -(PEG n3 (CH2) n4 CO )n5 - or HO2C(CH2) n1 CO-(AA4) n2 -(AA5) n6 - and provided that AA4 is selected from the group consisting of γGlu, εLys, β-Ala, γ-aminobutyric acid and 5-Ava, AA5 is selected from the group consisting of Gly, Ser, Thr, Asp, Glu, Aad, Lys, Orn, Dab and Dap] (II) a sequence in which one or more amino acids in the amino acid sequence represented by the above (I) are substituted, deleted, added and / or replaced; (III) a sequence having 90% or more homology with the amino acid sequence represented by the above (I) or (II); (IV) a pharmaceutically acceptable salt, solvate, chelate or non-covalent complex of the compound represented by the above formula I; (V) a prodrug based on the compound represented by the above formula I; (VI) a mixture optionally containing any one of the above (I), (II), (III), (IV) and / or (V).

[0010] In some specific embodiments of the present invention, the said n1 is any integer from 10 to 20, the said n2 is any integer from 1 to 5, the said n3 is any integer from 1 to 30, the said n4 is any integer from 1 to 5, the said n5 is any integer from 1 to 10, The n6 is an arbitrary integer from 1 to 10.

[0011] According to the present invention, there is further provided a method for producing the compound, including step 1 of producing a peptide resin by a solid-phase polypeptide synthesis method, and step 2 of obtaining the compound by acid decomposition and purification.

[0012] According to the present invention, GLP-1 Receptor and Gcg Receptor There is further provided the use of the following compound (I) and / or (II) in the production of a dual agonist: (I) The above-mentioned compound; (II) The compound produced by the above-mentioned production method.

[0013] According to the present invention, there is further provided the use of the following compound (I) and / or (II) in the production of a medicament or pharmaceutical composition for preventing and / or treating a disease: (I) The above-mentioned compound; (II) The compound produced by the above-mentioned production method.

[0014] In some specific embodiments of the present invention, the diseases include type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, metabolic syndrome, dyslipidemia, cognitive impairment, atherosclerosis, myocardial infarction, coronary heart disease, cardiovascular disease, stroke, inflammatory bowel syndrome and / or indigestion or gastric ulcer, liver fibrosis, and / or pulmonary fibrosis.

[0015] According to the present invention, there is further provided the use of the following compound (I) and / or (II) in the production of a medicament and / or pharmaceutical composition for treating type 2 diabetes with long-term continuous drug efficacy and / or preventing the deterioration of type 2 diabetes: (I) The above-mentioned compound; (II) The compound produced by the above-mentioned production method.

[0016] According to the present invention, there is further provided the use of the above-mentioned compound, or a compound produced by the above-mentioned production method, in the production of an agent and / or pharmaceutical composition for regulating the blood glucose level in the body. The regulation of the blood glucose level in the body includes a decrease in food intake, a decrease in β-cell apoptosis, an increase in pancreatic islet β-cell function, an increase in β-cell mass, and / or the restoration of the sensitivity of β-cells to glucose.

[0017] According to the present invention, there is further provided an agent or pharmaceutical composition comprising the following (I) and / or (II): (I) The above-mentioned compound; (II) A compound produced by the above-mentioned production method.

[0018] According to the present invention, there is further provided a method for regulating the blood glucose level in the body, which includes administering to a subject the following (I) and / or (II) and / or (III): (I) The above-mentioned compound; (II) A compound produced by the above-mentioned production method; (III) The above-mentioned agent or pharmaceutical composition.

Effects of the Invention

[0019] The present invention has beneficial effects including but not limited to the following effects. The compound represented by Formula I according to the present invention has a biological activity of lowering blood glucose level and reducing body weight Gcg / GLP-I dual agonist analog.

Modes for Carrying Out the Invention

[0020] The present invention discloses long-acting dual agonist compounds. Those skilled in the art can achieve the present invention by referring to the content of this specification and appropriately adjusting the process parameters. It should be noted that all similar substitutions and modifications are obvious to those skilled in the art and are all considered to be included in the present invention. The methods and uses according to the present invention have been described through preferred examples. However, it is obvious that those related can implement and apply the technology of the present invention by making corrections, appropriate changes and combinations to the methods and uses described in this specification without departing from the content, spirit and scope of the present invention.

[0021] An object of the present invention is to provide a GLP-1 / Gcg receptor dual agonist compound with a longer half-life, which is administered only once every two weeks or once every four weeks.

[0022] The present invention provides a long-acting dual agonist compound, which is a glucagon-like peptide-1 (GLP-1) receptor and glucagon (Gcg) receptor dual agonist compound, and its use.

[0023] To achieve the above object, first, the present invention provides a compound represented by formula I, a pharmaceutically acceptable salt, solvate, chelate, or non-covalent complex from this compound, a prodrug based on this compound, or any mixture of the above forms.

[0024] His-AA1-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Lys-Lys-Ala-Lys(R)-Glu-Phe-Val-Glu-Trp-Leu-Leu-AA2-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-AA3 [Formula I] [In formula I, AA1 is selected from the group consisting of Aib, Acpr, Acp, Acpe and Ach, AA2 is Glu or Ser, AA3 is NH2 or OH, R is HO2C(CH2) n1 CO-(AA4) n2 -(PEG n3 (CH2) n4 CO )n5 - or HO2C(CH2) n1 CO-(AA4) n2 -(AA5) n6 - and where n1 is any integer from 10 to 20, n2 is any integer from 1 to 5, n3 is any integer from 1 to 30, n4 is any integer from 1 to 5, n5 is any integer from 1 to 10, n6 is any integer from 1 to 10, AA4 is selected from the group consisting of γGlu, εLys, β-Ala, γ-aminobutyric acid, and 5-Ava, AA5 is selected from the group consisting of Gly, Ser, Thr, Asp, Glu, Aad, Lys, Orn, Dab, and Dap.]

[0025] According to the present invention, there is further provided a pharmaceutical composition of the compound according to the present invention, and the use of the pharmaceutical composition of the compound according to the present invention in the manufacture of a medicament for treating a disease.

[0026] Preferably, the pharmaceutical composition is used for manufacturing a medicament for treating at least one of diseases including type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, metabolic syndrome, dyslipidemia, cognitive impairment, atherosclerosis, myocardial infarction, coronary heart disease, cardiovascular disease, stroke, inflammatory bowel syndrome and / or indigestion or gastric ulcer, liver fibrosis, and pulmonary fibrosis.

[0027] Preferably, the pharmaceutical composition is used for manufacturing a medicament for treating type 2 diabetes with a long-term continuous therapeutic effect and / or preventing the exacerbation of type 2 diabetes.

[0028] Preferably, the pharmaceutical composition is used for manufacturing a medicament for reducing food intake, reducing β-cell apoptosis, increasing pancreatic islet β-cell function, increasing the β-cell mass, and / or restoring the sensitivity of β-cells to glucose. There is further provided a method of administering the compound to a subject for regulating blood glucose in vivo.

[0029] More details of what is referred to in the present invention will be described below in detail or may be understood from the examples of the present invention. Unless otherwise specified, the amounts of various components and reaction conditions used in this specification may be interpreted as meaning "substantially" or "approximately" in any case. Similarly, unless otherwise specified, the numerical parameters cited below and in the claims are all approximate parameters, and different numerical parameters may be obtained depending on the difference in the standard error under each experimental condition.

[0030] In this specification, if there is a discrepancy or doubt between the chemical structural formula and the chemical name of a compound, the compound defined by the chemical structural formula shall be regarded as accurate. The compounds described in this specification may contain one or more chiral centers, and / or double bonds, and may include such structures, and may also exist as stereoisomers such as double bond isomers (e.g., geometric isomers), optical enantiomers or diastereoisomers. Therefore, any chemical structure within the scope of this specification includes all possible enantiomers and diastereoisomers of this compound, even if it contains part or all of the above similar structures, including any pure stereoisomer (e.g., pure geometric isomer, pure enantiomer, or pure diastereoisomer) and any mixture of these isomers. These racemates and mixtures of stereoisomers can also be further separated into their constituent enantiomers or stereoisomers by various separation means or synthetic means of chiral molecules commonly used by those skilled in the art.

[0031] The compounds of Structural Formula I include, but are not limited to, optical isomers, racemates and / or other mixtures of these compounds. In the above cases, for example, a single enantiomer or diastereomer such as an optically active isomer can be obtained by an asymmetric synthesis method or a racemate separation method. The resolution of a racemate can be achieved by various methods such as conventional recrystallization using a separation promoter or chromatography. Also, the compounds of Formula I include cis and / or trans isomers having a double bond.

[0032] The compounds according to the present invention include, but are not limited to, the compounds represented by Formula I and all pharmaceutically usable forms thereof. Various pharmaceutically usable forms of these compounds include various pharmaceutically acceptable salts, solvates, complexes, chelates, non-covalent complexes, prodrugs based on the above substances, and any mixtures of these forms.

[0033] The compounds represented by Formula I according to the present invention are stable in nature and are hardly decomposed by dipeptidyl peptidase IV (DPP-IV) in the body. Gcg / GLP-I dual agonist analogs and have obvious blood glucose lowering and weight loss effects.

[0034] According to the present invention, there is further provided a production method including using Rink amide MBHA resin as a starting resin, producing a peptide resin by a polypeptide solid-phase synthesis method, then subjecting the peptide resin to acid decomposition to obtain a crude product, and finally purifying the crude product to obtain a pure product. Here, the step of producing a peptide resin by a polypeptide solid-phase synthesis method is to sequentially link the corresponding protected amino acids or fragments in the following sequence onto a carrier resin by a coupling solid-phase synthesis method to produce a peptide resin.

[0035] In the above production method, the usage amount of the Fmoc-protected amino acid or protected amino acid fragment is 1.2 to 6 times, preferably 2.5 to 3.5 times, the total molar number of the resin charged. In the above manufacturing method, the substitution value of the carrier resin is 0.2 to 1.0 mmol / g of resin, preferably 0.3 to 0.5 mmol / g of resin.

[0036] As a preferred embodiment of the present invention, in the coupling solid-phase synthesis method, after removing the Fmoc protecting group with the protected amino acid-resin obtained in the previous step, the next protected amino acid is subjected to a coupling reaction. The deprotection time for removing the Fmoc protecting group is 10 to 60 minutes, preferably 15 to 25 minutes. The coupling reaction time is 60 to 300 minutes, preferably 100 to 140 minutes.

[0037] A condensation reagent needs to be added to the coupling reaction. The condensation reagent is one selected from DIC (N,N-diisopropylcarbodiimide), N,N-dicyclohexylcarbodiimide, (benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate, 2-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate, benzotriazol-N,N,N',N'-tetramethyluronium hexafluorophosphate, or O-benzotriazol-N,N,N',N'-tetramethyluronium tetrafluoroborate, and preferably N,N-diisopropylcarbodiimide. The molar usage amount of the condensation reagent is 1.2 to 6 times the total molar number of amino groups in the amino resin, preferably 2.5 to 3.5 times.

[0038] An activating reagent needs to be added to the coupling reaction. The activating reagent is 1-hydroxybenzotriazole or N-hydroxy-7-azabenzotriazole, and preferably 1-hydroxybenzotriazole. The usage amount of the activating reagent is 1.2 to 6 times the total molar number of amino groups in the amino resin, preferably 2.5 to 3.5 times.

[0039] As a preferred embodiment of the present invention, the reagent for removing the Fmoc protecting group is a mixed solution of PIP / DMF (piperidine / N,N-dimethylformamide). The mixed solution contains 10 to 30% (V) of piperidine. The usage amount of the reagent for removing the Fmoc protecting group is 5 to 15 mL per gram of the amino resin, preferably 8 to 12 mL per gram of the amino resin.

[0040] Preferably, the peptide resin is acid-decomposed and the protecting groups of the resin and side chains are removed simultaneously to obtain a crude product.

[0041] More preferably, the acid decomposing agent used during the acid decomposition of the peptide resin is a mixed solvent of trifluoroacetic acid (TFA), 1,2-ethanedithiol (EDT), and water. The volume ratio of the mixed solvent is 80 to 95% of TFA, 1 to 10% of EDT, and the balance is water.

[0042] Even more preferably, the mixed solvent has a volume ratio of 89 to 91% of TFA, 4 to 6% of EDT, and the balance is water. Most preferably, the mixed solvent has a volume ratio of 90% of TFA, 5% of EDT, and the balance is water.

[0043] The usage amount of the acid decomposing agent is 4 to 15 mL of the acid decomposing agent per gram of the peptide resin, preferably 7 to 10 mL of the acid decomposing agent per gram of the peptide resin. The decomposition time by the acid decomposing agent is 1 to 6 hours under room temperature conditions, preferably 3 to 4 hours. Furthermore, the crude product is purified by high performance liquid chromatography and freeze-dried to obtain a pure product. The Japanese names corresponding to the English abbreviations mentioned in the present invention are shown in Table 1 below.

[0044]

Table 1

[0045] All raw materials and reagents used in the long-acting dual agonist compounds according to the present invention are commercially available. Hereinafter, the present invention will be further described with reference to examples. [Example 1] Production of Compound 1

[0046] The sequence of Compound 1 is represented by SEQ ID NO: 1. His-Aib-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Lys-Lys-Ala-Lys(AEEA-AEEA-γGlu-eicosanedioic acid)-Glu-Phe-Val-Glu-Trp-Leu-Leu-Glu-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH2 1. Synthesis of peptide resin

[0047] Using Rink Amide BHHA resin as the carrier resin, through Fmoc deprotection and coupling reactions, it was sequentially coupled with the protected amino acids shown in Table 2 below to produce a peptide resin. The protected amino acids corresponding to the protected amino acids used in this example are shown in Table 2 below.

[0048] [Table 2-1] [Table 2-2] (1) Linkage of the first protected amino acid of the main chain

[0049] Take 3 mmol of the first protected amino acid and 3 mmol of HOBt, dissolve them in an appropriate amount of DMF. Separately, take 3 mmol of DIC, and slowly add it to the DMF solution of the protected amino acid while stirring, and stir at room temperature for 30 minutes to react to obtain an activated protected amino acid solution, which was prepared for subsequent use.

[0050] 1 mmol of rink amide MBHA resin (substitution value is about 0.4 mmol / g) was taken, deprotected with 20% PIP / DMF solution for 25 minutes, washed, filtered, and the resin with Fmoc removed was obtained.

[0051] The activated first protected amino acid solution was added into the resin with Fmoc removed, and the coupling reaction was carried out for 60 - 300 minutes. After filtration and washing, the resin containing one protected amino acid was obtained. (2) Linkage of the 2nd - 39th protected amino acids of the main chain

[0052] Using the same method as the method for linking the first protected amino acid of the main chain, the corresponding 2nd - 39th protected amino acids were linked in sequence to obtain the resin containing 39 amino acids in the main chain. (3) Linkage of the first protected amino acid of the side chain

[0053] 3 mmol of the first protected amino acid of the side chain and 3 mmol of HOBt were dissolved in an appropriate amount of DMF. Separately, 3 mmol of DIC was taken and slowly added to the protected amino acid DMF solution while stirring, and stirred at room temperature for 30 minutes for reaction to obtain the activated protected amino acid solution.

[0054] 2.5 mmol of tetrakis(triphenylphosphine)palladium and 25 mmol of phenylsilane were taken, dissolved in an appropriate amount of dichloromethane, deprotected for 4 hours, filtered, washed, and the resin with Alloc removed was obtained and prepared for subsequent use.

[0055] The activated first protected amino acid solution of the side chain was added into the resin with Alloc removed, and the coupling reaction was carried out for 60 - 300 minutes. After filtration and washing, the resin containing the first protected amino acid of the side chain was obtained. (4) Linkage of the protected amino acids of the side chain

[0056] Using the same method as the method for linking the first protected amino acid of the main chain, the corresponding protected amino acids and monoprotected fatty acids of the side chain were linked in sequence to obtain the peptide resin. 2. Production of the crude product

[0057] Take the peptide resin described above and add it to a cleavage reagent with a volume ratio of TFA:water:EDT = 95:5:5 (10 mL of cleavage reagent per gram of resin). Stir to make it uniform, stir and react at room temperature for 3 hours. Filter the reaction mixture through a sand core funnel, collect the filtrate, wash the resin three times with a small amount of TFA, combine the filtrates, concentrate under reduced pressure, add diethyl ether (anhydrous) to precipitate, wash the precipitate three times with diethyl ether (anhydrous), and dry in vacuo to obtain a white powder, i.e., the crude product. 3. Production of the pure product

[0058] Add water to the crude product described above, stir, adjust the pH to 8.0 with aqueous ammonia until completely dissolved, filter the solution through a 0.45 μm mixed microporous membrane for purification, and prepare it for subsequent use.

[0059] For purification, high performance liquid chromatography was used. 10 μm reversed-phase C18 was used as the chromatographic packing for purification. A 0.1% TFA / aqueous solution - 0.1% TFA / acetonitrile solution was used as the mobile phase system. The flow rate of a 30 mm × 250 mm chromatographic column was set at 20 mL / min. Gradient elution was used and purification was carried out by the cyclic sample loading method. Take the solution of the crude product, inject it into the chromatographic column, start the mobile phase elution, collect the main peak, evaporate the acetonitrile, and obtain a concentrated solution of the purified intermediate.

[0060] The refined intermediate concentrate was filtered through a 0.45 μm filter membrane and prepared for subsequent use. Salt exchange was carried out by high performance liquid chromatography. The mobile phase system was 1% acetic acid / water solution - acetonitrile, the chromatography packing for purification was 10 μm reversed-phase C18, and the flow rate of a 30 mm × 250 mm chromatography column was set at 20 mL / min (the flow rate can be adjusted accordingly according to the specifications of the chromatography column). It was injected into the chromatography column by gradient elution and cyclic sample loading methods, the mobile phase elution was started, the chromatograph was collected, the change in absorbance was observed, the main peak of salt exchange was collected, the purity of the liquid phase was detected, the main peak solutions of salt exchange were combined and concentrated under reduced pressure to obtain a pure acetic acid aqueous solution, which was freeze-dried to obtain 0.35 g of a pure product. The purity was 97.9%, the total yield was 7.0%, and the molecular weight was 5012.6 (100% M+H). [Example 2] Preparation of Compound 2

[0061] The sequence of Compound 2 is shown in SEQ ID NO: 2. His-Aib-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Lys-Lys-Ala-Lys(PEG5CH2CO-γGlu-eicosanedioic acid)-Glu-Phe-Val-Glu-Trp-Leu-Leu-Glu-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH2

[0062] The manufacturing method was the same as in Example 1, and the protected amino acids shown in Table 3 below were used.

Table 3-1

Table 3-2

[0063] 0.43 g of a pure product was obtained. The purity was 97.0%, the total yield was 8.6%, and the molecular weight was 4999.6 (100% M+H). [Example 3] Preparation of Compound 3

[0064] The sequence of Compound 3 is represented by SEQ ID NO: 3. His-Aib-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Lys-Lys-Ala-Lys(Gly-Gly-Ser-Gly-Ser-Gly-γGlu-eicosanedioic acid-γGlu-eicosanedioic acid)-Glu-Phe-Val-Glu-Trp-Leu-Leu-Glu-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH2

[0065] The production method was carried out in the same manner as in Example 1, using the protected amino acids shown in Table 4 below. [Table 4-1] [Table 4-2]

[0066] 0.33 g of a pure product was obtained. The purity was 93.4%, the total yield was 6.4%, and the molecular weight was 5124.6 (100% M+H). [Example 4] Production of Compound 4

[0067] The sequence of Compound 4 is represented by SEQ ID NO: 4. His-Aib-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Lys-Lys-Ala-Lys(Gly-Gly-Glu-Gly-Glu-Gly-γGlu-eicosanedioic acid)-Glu-Phe-Val-Glu-Trp-Leu-Leu-Glu-Gly-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH2

[0068] The production method was carried out in the same manner as in Example 1, using the protected amino acids shown in Table 5 below. [Table 5-1] [Table 5-2]

[0069] 0.30 g of pure product was obtained. The purity was 96.5%, the total yield was 5.8%, and the molecular weight was 5208.7 (100% M+H). [Example 5] Production of Compound 5

[0070] The sequence of Compound 5 is shown by SEQ ID NO: 5. His-Aib-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Lys-Lys-Ala-Lys(AEEA-AEEA-γGlu-eicosanedioic acid)-Glu-Phe-Val-Glu-Trp-Leu-Leu-Glu-Gly-Gly-Pro-Ser-Ser-Gly-NH2

[0071] The production method was the same as that in Example 1. The molecular weight was 4563.1. [Example 6] GLP-1 Activity Measurement 1. Measurement Method

[0072] GLP-1R can activate the intracellular adenylate cyclase pathway under the stimulation of a specific agonist, increase the level of cAMP, and as a result, insulin production and release are brought about. By stimulating a cell line stably transfected with GLP-1R with a test substance, the intracellular cAMP level is rapidly increased, and the relative light unit (RLU) after stimulating the cells at each dosage is measured by chemiluminescence method, and the EC50 of the agonist is calculated. This activity measurement method is currently an activity detection method for GLP-1 receptor agonists commonly used at home and abroad.

[0073] The CHO-K1 cell line stably expressing GLP-1R was used, and the stably transfected cells were stimulated with agonists at different concentrations. The relative light unit after stimulating the cells at each dosage was measured, and by using OXM3 as a control sample, the relative biological activity of the agonist was obtained. 2. Measurement Results

[0074] The measurement results are shown in Table 6 below. [Table 6] As a result of the test, the GLP-1 activity of the novel Gcg / GLP-1 dual agonist analog was improved by 15% to over 159% compared with OXM3. [Example 7] Measurement of Gcg Activity 1. Measurement Method

[0075] GcgR can activate the intracellular adenylate cyclase pathway under the stimulation of a specific agonist, increase the level of cAMP, and as a result, bring about the production and release of insulin. GcgR By stimulating the cell line stably transfected with Gcg with the test substance, the intracellular cAMP level can be rapidly increased. After stimulating the cells at each dosage, the relative light unit (RLU) is measured by chemiluminescence method, and the EC50 of the agonist is calculated. This activity measurement method is currently commonly used at home and abroad

[0076] as an activity detection method for receptor agonists. 2. Measurement Results

[0077] The measurement results are shown in Table 7 below. [Table 7] As a result of the test, the Gcg activities of the novel Gcg / GLP-1 dual agonist analogs 1 to 3 were almost the same as the activity of OXM3. [Example 8] Measurement of Preliminary Pharmacokinetic Characteristics

[0078] Cynomolgus monkeys were used as experimental animals. Two male cynomolgus monkeys were assigned to each compound administration group and administered subcutaneously at a dose of 0.2 mg / kg. Blood samples were collected from the vein before administration (0 hours) and at 1 hour, 2 hours, 3 hours, 4 hours, 8 hours, 12 hours, 18 hours, 24 hours, 48 hours, 96 hours, 144 hours, and 168 hours after administration. The plasma samples were centrifuged. The blood drug concentration of the corresponding compound in each Cynomolgus monkey plasma sample was measured by liquid chromatography-mass spectrometry. The half-life of the compound at subcutaneous (SC) administration is shown in Table 8 below.

[0079]

Table 8

[0080] The pharmacokinetic properties of the modified compound (Compound 2) with the highest biological activity were measured. As a result, the half-life of Compound 2 was 140.2 hours, which reached approximately 280 hours when converted to the half-life of humans, and was much higher than the half-life of OXM3 (120 hours).

[0081] The long-acting dual agonist compounds according to the present invention have been described in detail above. In this specification, the principles and embodiments of the present invention have been described using specific examples, but the description of the above examples is only used to assist in understanding the method and the essence of the present invention. Here, it should be noted that those skilled in the art can make various improvements and modifications to the present invention without departing from the principles of the present invention, and these improvements and modifications are also included in the scope of the claims of the present invention.

Claims

1. A compound comprising any one of the following (I) or (II) or (III) or (IV) and / or (V) and / or (VI): (I) An amino acid sequence represented by the following formula I; His-AA1-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Lys-Lys-Ala-Lys(R)-Glu-Phe-Val-Glu-Trp-Leu-Leu-AA2-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-AA3 [Formula I] [In Formula I, AA1 is selected from the group consisting of Aib, Acpr, Acp, Acpe and Ach, AA2 is Glu or Ser, and AA3 is NH ２ or OH, R is HO ２ C(CH ２ ) ｎ１ CO-(AA4) ｎ２ -(PEG ｎ３ (CH2) ｎ４ CO ）ｎ５ - or HO ２ C(CH ２ ) ｎ１ CO-(AA4) ｎ２ -(AA5) ｎ６ - and provided that AA4 is selected from the group consisting of γGlu, εLys, β-Ala, γ-aminobutyric acid and 5-Ava, AA5 is selected from the group consisting of Gly, Ser, Thr, Asp, Glu, Aad, Lys, Orn, Dab and Dap]] (II) A sequence in which one or more amino acids are substituted, deleted, added and / or replaced in the amino acid sequence represented by (I) above; (III) A sequence having 90% or more homology with the amino acid sequence represented by (I) or (II) above; (IV) A pharmaceutically acceptable salt, solvate, chelate or non-covalent complex of the compound represented by the above formula I; (V) A prodrug based on the compound represented by the above formula I; (VI) A mixture optionally containing any one of (I), (II), (III), (IV) and / or (V) above.

2. Wherein n1 is any integer from 10 to 20, n2 is any integer from 1 to 5, n3 is any integer from 1 to 30, n4 is any integer from 1 to 5, n5 is any integer from 1 to 10, n6 is any integer from 1 to 10, The compound according to claim 1, characterized in that.

3. Step 1 of producing a peptide resin by the solid-phase synthesis method of polypeptides, A method for producing a compound according to claim 1 or 2, comprising step 2 of obtaining the compound by acid decomposition and purification.

4. Use of the following compounds (I) and / or (II) in the production of a GLP-1 and Gcg dual agonist: (I) The compound according to claim 1 or 2; (II) A compound produced by the production method according to claim 3.

5. Use of a compound of the following (I) and / or (II) in the manufacture of a medicament or pharmaceutical composition for preventing and / or treating a disease: (I) The compound according to claim 1 or 2; (II) The compound produced by the production method according to claim 3.

6. The use according to claim 5, wherein the disease is type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, metabolic syndrome, dyslipidemia, cognitive impairment, atherosclerotic disease, myocardial infarction, coronary heart disease, cardiovascular disease, stroke, inflammatory bowel syndrome and / or indigestion or gastric ulcer, liver fibrosis, and / or pulmonary fibrosis.

7. Use of a compound of the following (I) and / or (II) in the manufacture of a medicament and / or pharmaceutical composition for treating type 2 diabetes with a long-term continuous medicinal effect and / or preventing the deterioration of type 2 diabetes: (I) The compound according to claim 1 or 2; (II) The compound produced by the production method according to claim 3.

8. Use of the compound according to claim 1 or 2, or the compound produced by the production method according to claim 3, in the manufacture of a medicament and / or pharmaceutical composition for regulating the blood glucose level in the body, wherein regulating the blood glucose level in the body is a decrease in food intake, a decrease in β-cell apoptosis, an increase in pancreatic islet β-cell function, an increase in β-cell mass, and / or a restoration of the sensitivity of β-cells to glucose.

9. (I) The compound according to claim 1 or 2, and / or (II) A medicament or pharmaceutical composition characterized by comprising the compound produced by the production method according to claim 3.

10. (I) The compound according to claim 1 or 2, and / or (II) The compound produced by the production method according to claim 3, and / or (III) A method for regulating the blood glucose level in the body, characterized by comprising administering the medicament or pharmaceutical composition according to claim 9 to a subject.

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