GIP agonist compound

Abstract

The invention relates to the field of medicine synthesis, and discloses a long-acting GIP compound. The long-acting GIP compound is used for preparing a medicine composition for treating diseases, and the application of the medicine composition in preparing a medicine for treating at least one of the following diseases including diabetes mellitus, obesity, metabolic syndrome, dyslipidemia, atherosclerosis, hepatic fibrosis diseases, pulmonary fibrosis and the like.

Description

Technical Field

[0001] The present invention relates to a GIP agonist compound and its use. The compound is a modified glucagon agonist compound. Background Art

[0002] Human glucose-dependent insulinotropic polypeptide (GIP) is the first incretin discovered, and the currently known incretins are GIP and GLP-1. In normal human beings, the insulin secretion stimulated by the incretin effect (after a meal) accounts for about 70% of the total insulin secretion in the whole body. GIP accounts for 2 / 3 in the whole incretin effect, with a proportion much higher than that of GLP-1.

[0003] GIP is a 42-amino acid gastrointestinal regulatory peptide, which plays a physiological role in glucose homeostasis by stimulating insulin secretion from pancreatic β-cells and protecting pancreatic β-cells in the presence of glucose. The primary structure of GIP is: Tyr-Ala-Glu-Gly-Thr-Phe-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys-Gly-Lys-Lys-Asn-Asp-Trp-Lys-His-Asn-Ile-Thr-Gln-OH

[0004] GIP is widely distributed in different organ tissues, such as the pancreas, stomach, adipose tissue, lung, heart, bone, vascular endothelium, and some parts of the brain tissue. It belongs to the incretin and is an important metabolic hormone in the human body, playing an important physiological function in the human body, mainly stimulating insulin release and glucagon secretion, inhibiting gastric acid and pepsin secretion, and inhibiting gastric motility and emptying, etc.

[0005] In normal people, the half-life of GIP is about 7 minutes, and even shorter in type 2 diabetes mellitus (T2DM) patients. Because the expression level and activity of DPP-4 enzyme in T2DM patients increase, the half-life of GIP is shortened to 5 minutes. Due to the too short half-life, the clinical application of GIP is limited. The purpose of the present invention is to provide a long-acting GIP agonist compound for patients, reduce the dosing frequency, and provide a new choice for patients. Summary of the Invention

[0006] The present invention provides a GIP agonist compound and its use. The compound is a glucagon GIP agonist compound.

[0007] In the first aspect of the embodiments of the present invention, a GIP agonist compound is provided: Tyr-Aib-Glu-Gly-Thr-AA1-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys((CO(CH2) n1 PEG n2 ) n3 -(AA2) n4 -CO(CH2) n5 -COOH)-AA3 Structural formula Ⅰ Wherein: AA1 is selected from any one of Phe, αMePhe, and αMePhe(2F); AA2 is selected from any one of γGlu, δAad, εApm, and ζAsu; AA3 is selected from any one of amino group and hydroxyl group; n1 = 1 - 5, n2 = 1 - 30, n3 = 0 or = 1 - 5, n4 = 0 or = 1 - 5, n5 = 10 - 30;

[0008] Optionally, the GIP agonist is characterized in that AA1 is selected as Phe, AA2 is selected as γGlu, AA3 is selected as amino group, n1 = 1 - 5, n2 = 1 - 30, n3 = 0 or = 1 - 5, n4 = 0 or = 1 - 5, n5 = 10 - 30; Preferably, AA1 is selected as Phe, AA2 is selected as γGlu, AA3 is selected as amino group, n1 = 1, n2 = 5, n3 = 1, n4 = 1, n5 = 18;

[0009] Optionally, the GIP agonist is characterized in that AA1 is selected as αMePhe, AA2 is selected as γGlu, AA3 is selected as amino group, n1 = 1 - 5, n2 = 1 - 30, n3 = 0 or = 1 - 5, n4 = 0 or = 1 - 5, n5 = 10 - 30; Preferably, AA1 is selected as αMePhe, AA2 is selected as γGlu, AA3 is selected as amino group, n1 = 1, n2 = 5, n3 = 1, n4 = 1, n5 = 18;

[0010] Optionally, the GIP agonist is characterized in that AA1 is selected as αMePhe, AA2 is selected as δAad, AA3 is selected as amino group, n1 = 1 - 5, n2 = 1 - 30, n3 = 0 or = 1 - 5, n4 = 0 or = 1 - 5, n5 = 10 - 30; Preferably, AA1 is selected as αMePhe, AA2 is selected as δAad, AA3 is selected as amino group, n1 = 1, n2 = 5, n3 = 1, n4 = 1, n5 = 18;

[0011] Optionally, the GIP agonist is characterized in that AA1 is selected as αMePhe(2F), AA2 is selected as γGlu, AA3 is selected as amino, n1 = 1 - 5, n2 = 1 - 30, n3 = 0 or = 1 - 5, n4 = 0 or = 1 - 5, n5 = 10 - 30; preferably, AA1 is selected as αMePhe(2F), AA2 is selected as γGlu, AA3 is selected as amino, n1 = 1, n2 = 5, n3 = 1, n4 = 1, n5 = 18;

[0012] Optionally, the GIP agonist is characterized in that AA1 is selected as αMePhe(2F), AA2 is selected as δAad, AA3 is selected as amino, n1 = 1 - 5, n2 = 1 - 30, n3 = 0 or = 1 - 5, n4 = 0 or = 1 - 5, n5 = 10 - 30; preferably, AA1 is selected as αMePhe(2F), AA2 is selected as δAad, AA3 is selected as amino, n1 = 1, n2 = 5, n3 = 1, n4 = 1, n5 = 18;

[0013] Optionally, the above GIP agonist compound is characterized in that the GIP agonist compound comprises a medicinal salt, chelate or non-covalent complex formed by the compound, and a precursor of the compound, or any mixture of the above forms.

[0014] The second aspect of the embodiments of the present invention provides an application of a GIP agonist compound as described in any one of the above first aspects in the preparation of a pharmaceutical composition for treating diseases.

[0015] Optionally, the pharmaceutical composition for treating diseases is characterized in that the pharmaceutical composition for treating diseases is used for at least one of the following diseases, and the diseases include diabetes, obesity, metabolic syndrome, dyslipidemia, atherosclerosis, liver fibrosis diseases, pulmonary fibrosis and other diseases.

[0016] Unless otherwise indicated, the quantities and reaction conditions used herein to represent different components can be interpreted as "substantially" and "approximately" in any case. Correspondingly, unless specifically specified, the numerical parameters cited in the following and the claims are approximate parameters, and due to different standard errors under their respective experimental conditions, different numerical parameters may be obtained.

[0017] In this article, when there are differences or ambiguities between the chemical structural formula and the chemical name of a compound, the compound is defined precisely by its chemical structural formula. The compounds described herein may contain one or more chiral centers, and / or double bonds and structures of this kind, and there may also exist stereoisomers, including isomers of double bonds (such as geometric isomers), enantiomers or diastereoisomers. Accordingly, any chemical structure within the scope described herein, whether it contains the above-mentioned similar structures in part or in whole, includes all possible enantiomers and diastereoisomers of this compound, including any pure stereoisomer (such as a pure geometric isomer, a pure enantiomer or a pure diastereoisomer) and any mixture of these isomers. These racemates and mixtures of stereoisomers can also be further separated into their component enantiomers or stereoisomers by those skilled in the art using separation techniques or chiral molecular synthesis methods.

[0018] The compounds of Structural Formula I include, but are not limited to, the optical isomers, racemates and / or other mixtures of these compounds. In the above cases, a single enantiomer or diastereoisomer, such as an optically active isomer, can be obtained by asymmetric synthesis or racemate resolution methods. Racemate resolution can be achieved by different methods, such as conventional recrystallization with a resolving agent or by chromatography. In addition, the compounds of Structural Formula I also include cis and / or trans isomers with double bonds.

[0019] The compounds described in the present invention include, but are not limited to, the compounds shown in Structural Formula I and all their different pharmaceutically available forms. These different pharmaceutically available forms of the compounds include various pharmaceutically acceptable salts, solvates, complexes, chelates, non-covalent complexes, prodrugs based on the above substances and any mixture of the above forms.

[0020] The compound shown in Structural Formula I provided by the present invention has stable properties and is a highly active GIP agonist compound, with significant hypoglycemic and weight loss effects. Detailed implementation mode

[0021] The present invention discloses a GIP agonist compound and its uses, and those skilled in the art can draw on the content of this article and appropriately modify relevant parameters to achieve them. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are all regarded as included in the present invention. The methods of the present invention have been described through preferred embodiments, and those skilled in the art can obviously make changes or appropriate alterations and combinations to the compounds and preparation methods described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0022] The Chinese names corresponding to the English abbreviations involved in the present invention are shown in the following table: English Abbreviation Chinese Name English Abbreviation Chinese Name Fmoc 9-Fluorenylmethyloxycarbonyl OtBu tert-Butoxy tBu tert-Butyl Boc tert-Butyloxycarbonyl Trt Triphenylmethyl Pbf (2,3-Dihydro-2,2,4,6,7-pentamethylbenzofuran-5-yl)sulfonyl Ala Alanine Leu Leucine Arg Arginine Lys Lysine Asn Asparagine Phe Phenylalanine Asp Aspartic Acid Pro Proline Cys Cysteine Ser Serine Gln Glutamine Thr Threonine Glu Glutamic Acid Trp Tryptophan Gly Glycine Tyr Tyrosine His Histidine Val Valine Ile Isoleucine Ada 2-Aminoadipic Acid Aib Aminoisobutyric Acid Apm 2-Aminoheptanedioic Acid 5-Ava 5-Aminovaleric Acid Asu 2-Aminooctanedioic Acid

[0023] Preparation of the compound in Example 1

[0024] The preparation method includes: preparing a peptide resin by solid-phase peptide synthesis, subjecting the peptide resin to acidolysis to obtain a crude product, and finally purifying the crude product to obtain a pure product; wherein the steps of preparing the peptide resin by solid-phase peptide synthesis are to sequentially connect the corresponding protected amino acids or fragments in the polypeptide sequence onto the carrier resin by solid-phase coupling synthesis to prepare the peptide resin:

[0025] In the above preparation method, the amount of the Fmoc-protected amino acid or protected amino acid fragment used is 1.2 to 6 times the total molar amount of the charged resin; preferably 2.5 to 3.5 times.

[0026] In the above preparation method, the substitution value of the carrier resin is 0.2 to 1.0 mmol / g resin, and the preferred substitution value is 0.3 to 0.5 mmol / g resin.

[0027] As a preferred embodiment of the present invention, the solid-phase coupling synthesis method is: after the protected amino acid-resin obtained from the previous reaction is deprotected from the Fmoc protecting group, it is then coupled with the next protected amino acid. The deprotection time for removing the Fmoc protection is 10 to 60 minutes, preferably 15 to 25 minutes. The coupling reaction time is 60 to 300 minutes, preferably 100 to 140 minutes.

[0028] The coupling reaction needs to add a condensation reagent, and the condensation reagent is selected from one of DIC (N,N-diisopropylcarbodiimide), N,N-dicyclohexylcarbodiimide, benzotriazol-1-yl-oxytripyrrolidinophosphonium 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; preferably N,N-diisopropylcarbodiimide. The molar amount of the condensation reagent used is 1.2 to 6 times the total molar amount of the amino groups in the amino resin, preferably 2.5 to 3.5 times.

[0029] The coupling reaction needs to add an activation reagent, and the activation reagent is selected from 1-hydroxybenzotriazole or N-hydroxy-7-azabenzotriazole, preferably 1-hydroxybenzotriazole. The amount of the activation reagent used is 1.2 to 6 times the total molar amount of the amino groups in the amino resin, preferably 2.5 to 3.5 times.

[0030] As a preferred embodiment of the present invention, the reagent for removing the Fmoc protection is a mixed solution of PIP / DMF (piperidine / N,N-dimethylformamide), and the piperidine in the mixed solution is 10-30% (V). The dosage of the reagent for removing the Fmoc protection is 5-15 mL per gram of amino resin, preferably 8-12 mL per gram of amino resin.

[0031] Preferably, the peptide resin is simultaneously deprotected from the resin and side chain protecting groups by acidolysis to obtain a crude product:

[0032] More preferably, the acidolysis agent used for 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: TFA is 80-95%, EDT is 1-10%, and the balance is water.

[0033] Even more preferably, the volume ratio of the mixed solvent is: TFA is 89-91%, EDT is 4-6%, and the balance is water. Most preferably, the volume ratio of the mixed solvent is: TFA is 90%, EDT is 5%, and the balance is water.

[0034] The dosage of the acidolysis agent is 4-15 mL of acidolysis agent per gram of peptide resin; preferably, 7-10 mL of acidolysis agent is required per gram of peptide resin.

[0035] The time for cleavage using the acidolysis agent is 1-6 hours at room temperature, preferably 3-4 hours.

[0036] Furthermore, the crude product is purified by high performance liquid chromatography and freeze-dried to obtain a pure product.

[0037] 1. Synthesis of peptide resin

[0038] Using Rink Amide BHHA resin as the carrier resin, through de-Fmoc protection and coupling reaction, it is successively coupled with the protected amino acids corresponding to the polypeptide sequence to prepare the peptide resin.

[0039] (1) Incorporating the first protected amino acid of the main chain

[0040] Take 0.03 mol of the first protected amino acid and 0.03 mol of HOBt, and dissolve them in an appropriate amount of DMF; separately take 0.03 mol of DIC, and slowly add it to the protected amino acid DMF solution under stirring, and stir and react at room temperature for 30 minutes to obtain an activated protected amino acid solution for standby.

[0041] Take 0.01 mol of Rink amide MBHA resin (substitution value about 0.4 mmol / g), and deprotect it with 20% PIP / DMF solution for 25 minutes, wash and filter to obtain the de-Fmoc resin.

[0042] Add the activated solution of the first protected amino acid to the resin from which Fmoc has been removed, carry out the coupling reaction for 60 - 300 minutes, filter and wash to obtain the resin containing one protected amino acid.

[0043] (2) Incorporate the protected amino acids of the main chain

[0044] Using the same method as above for incorporating the first protected amino acid of the main chain, sequentially incorporate the protected amino acids corresponding to the respective polypeptide sequences to obtain the resin containing the amino acids of the main chain.

[0045] (3) Incorporate the first protected amino acid of the side chain

[0046] Take 0.03 mol of the first protected amino acid of the side chain and 0.03 mol of HOBt, dissolve them in an appropriate amount of DMF; separately take 0.03 mol of DIC and slowly add it to the DMF solution of the protected amino acid with stirring, and stir and react at room temperature for 30 minutes to obtain the activated solution of the protected amino acid.

[0047] Take 2.5 mmol of tetrakis(triphenylphosphine)palladium and 25 mmol of phenylsilane, dissolve them in an appropriate amount of dichloromethane, carry out deprotection for 4 hours, filter and wash to obtain the resin with Alloc removed for standby.

[0048] Add the activated solution of the first protected amino acid of the side chain to the resin from which Alloc has been removed, carry out the coupling reaction for 60 - 300 minutes, filter and wash to obtain the resin containing the first protected amino acid of the side chain.

[0049] (4) Incorporate other protected amino acids or mono - protected fatty acids of the side chain

[0050] Using the same method as above for incorporating the first protected amino acid of the main chain, sequentially incorporate the corresponding protected amino acids and mono - protected fatty acids of the side chain to obtain the peptide resin.

[0051] 2. Preparation of the crude product

[0052] Take the above - mentioned peptide resin, add the cleavage reagent with a volume ratio of TFA︰water︰EDT = 95︰5︰5 (10 mL of the cleavage reagent per gram of resin), stir evenly, stir and react at room temperature for 3 hours, filter the reaction mixture using a sintered glass funnel, collect the filtrate, wash the resin with a small amount of TFA 3 times, combine the filtrates and concentrate under reduced pressure, add anhydrous ether for precipitation, and then wash the precipitate with anhydrous ether 3 times, and drain to obtain an off - white powder, which is the crude product.

[0053] 3. Preparation of the pure product

[0054] Take the above - mentioned crude product, add water and stir, adjust the pH to 8.0 with ammonia water until completely dissolved, filter the solution through a 0.45 μm mixed microporous filter membrane for purification and standby;

[0055] Purification was carried out by high performance liquid chromatography. The chromatographic packing material for purification was 10 μm reversed-phase C18. The mobile phase system was 0.1% TFA / aqueous solution - 0.1% TFA / acetonitrile solution. The flow rate of the 30 mm * 250 mm chromatographic column was 20 mL / min. Gradient elution was used, and cyclic injection purification was carried out. The crude product solution was loaded onto the chromatographic column, the mobile phase elution was started, and after collecting the main peak and evaporating the acetonitrile, a purified intermediate concentrate was obtained.

[0056] The purified intermediate concentrate was filtered through a 0.45 μm filter membrane for standby. Salt exchange was carried out by high performance liquid chromatography. The mobile phase system was 1% acetic acid / aqueous solution - acetonitrile. The chromatographic packing material for purification was 10 μm reversed-phase C18. The flow rate of the 30 mm * 250 mm chromatographic column was 20 mL / min (the flow rate can be adjusted according to different specifications of the chromatographic column); gradient elution was used, and the cyclic loading method was used to load onto the chromatographic column. The mobile phase elution was started, the chromatogram was collected, the change in absorbance was observed, the main peak of salt exchange was collected and the purity was detected by analytical liquid chromatography. The main peak solutions of salt exchange were combined and concentrated under reduced pressure to obtain a pure acetic acid aqueous solution, and freeze-dried to obtain pure peptide.

[0057] The following compounds were synthesized by the above method: Compound Sequence Structure Positive Control GIP Tyr-Ala-Glu-Gly-Thr-Phe-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys-Gly-Lys-Lys-Asn-Asp-Trp-Lys-His-Asn-Ile-Thr-Gln-OH Compound 1 <![CDATA[Tyr-Aib-Glu-Gly-Thr-Phe-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys(COCH2PEG5-γGlu-eicosanedioic acid)-NH2]]> Compound 2 <![CDATA[Tyr-Aib-Glu-Gly-Thr-Phe-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys(COCH2PEG5-δAad-eicosanedioic acid)-NH2]]> Compound 3 <![CDATA[Tyr-Aib-Glu-Gly-Thr-Phe-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys(COCH2PEG5-εApm-eicosanedioic acid)-NH2]]> Compound 4 <![CDATA[Tyr-Aib-Glu-Gly-Thr-Phe-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys(COCH2PEG5-ζAsu-eicosanedioic acid)-NH2]]> Compound 5 <![CDATA[Tyr-Aib-Glu-Gly-Thr-αMePhe-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys(COCH2PEG5-γGlu-eicosanedioic acid)-NH2 <!-- 5 -->]]> Compound 6 <![CDATA[Tyr-Aib-Glu-Gly-Thr-αMePhe-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys(COCH2PEG5-δAad-eicosanedioic acid)-NH2]]> Compound 7 <![CDATA[Tyr-Aib-Glu-Gly-Thr-αMePhe-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys(COCH2PEG5-εApm-eicosanedioic acid)-NH2]]> Compound 8 <![CDATA[Tyr-Aib-Glu-Gly-Thr-αMePhe-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys(COCH2PEG5-ζAsu-eicosanedioic acid)-NH2]]> Compound 9 <![CDATA[Tyr-Aib-Glu-Gly-Thr-αMePhe(2F)-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys(COCH2PEG5-γGlu-eicosanedioic acid)-NH2]]> Compound 10 <![CDATA[Tyr-Aib-Glu-Gly-Thr-αMePhe(2F)-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys(COCH2PEG5-δAad-eicosanedioic acid)-NH2]]> Compound 11 <![CDATA[Tyr-Aib-Glu-Gly-Thr-αMePhe(2F)-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys(COCH2PEG5-εApm-icosanedioic acid)-NH2]]> Compound 12 <![CDATA[Tyr-Aib-Glu-Gly-Thr-αMePhe(2F)-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys(COCH2PEG5-ζAsu-eicosanedioic acid)-NH2]]> Compound 13 Tyr-Aib-Glu-Gly-Thr-Phe-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys(AEEA-AEEA-γGlu-eicosanedioic acid)-NH2 Compound 14 Tyr-Aib-Glu-Gly-Thr-αMePhe-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys(AEEA-AEEA-γGlu-eicosanedioic acid)-NH2 Compound 15 Tyr-Aib-Glu-Gly-Thr-αMePhe(2F)-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys(AEEA-AEEA-γGlu-eicosanedioic acid)-NH2

[0058] Example 2 Determination of Activity

[0059] 1. GIP Activity Determination Method

[0060] Under the stimulation of its specific agonist, GIP-R can activate the intracellular adenylate cyclase pathway, increase the cAMP level, and ultimately lead to the production and release of insulin. By stimulating the cell line stably transfected with GIP-R with the test substance, the intracellular cAMP level of the cells can be rapidly increased. The relative light units (RLU) after stimulating the cells at each dose are measured by chemiluminescence method, and then the EC50 of the agonist is calculated. This activity determination method is a commonly used method for detecting the activity of GIP receptor agonists at home and abroad.

[0061] The CHO-K1 cell line stably expressing GIP-R was used. The stably transfected cells were stimulated with agonists at different concentrations. By measuring the relative light units after stimulating the cells at each dose, the EC 50 value was calculated.

[0062] 2. Determination Results

[0063] The determination results are shown in the following table: Compound <![CDATA[GIP

EC 50 (pmol)

[0064] The experimental results show that the activities of all compounds GIP in the examples are at the pmol level, meeting the activity requirements of long-acting compounds.

[0065] Example 3 Determination of Preliminary Pharmacokinetic Properties

[0066] Among the above compounds, the principle of long-acting modification is basically the same. Therefore, the two compounds with the best activity were selected for the verification test of preliminary pharmacokinetic properties.

[0067] The test animals were cynomolgus monkeys, male. Two cynomolgus monkeys were used for each compound. The drug was administered subcutaneously at a dose of 0.1 mg / kg. Blood was collected intravenously before drug administration (0 h) and at 1 h, 2 h, 3 h, 4 h, 8 h, 12 h, 18 h, 24 h, 48 h, 96 h, 144 h, and 168 h after drug administration. Plasma samples were separated by centrifugation, and the plasma concentrations of the corresponding compounds in the plasma samples were determined by liquid chromatography-mass spectrometry. The subcutaneous (SC) half-lives of the compounds are shown in the following table: Compound <![CDATA[t 1 / 2 (h)]]> Compound 5 77.2 Compound 9 86.9

[0068] The results of the preliminary pharmacokinetic properties experiment in cynomolgus monkeys showed that the subcutaneous half-lives of Compound 5 and Compound 9 were both greater than 70 hours, fully meeting the requirements of long-acting drug administration.

Claims

1. A GIP agonist compound having the structural formula I: Tyr-Aib-Glu-Gly-Thr-AA1-Ile-Ser-Asp-Tyr-Ser-Ile-Ala-Met-Asp-Lys-Ile-His-Gln-Gln-Asp-Phe-Val-Asn-Trp-Leu-Leu-Ala-Gln-Lys((CO(CH2) n1 PEG n2 ) n3 -(AA2) n4 -CO(CH2) n5 -COOH)-AA3 Structural formula Ⅰ Wherein: AA1 is selected from any one of Phe, αMePhe, αMePhe(2F); AA2 is selected from any one of γGlu, δAad, εApm, ζAsu; AA3 is selected from any one of amino group, hydroxyl group; n1 = 1 - 5, n2 = 1 - 30, n3 = 0 or = 1 - 5, n4 = 0 or = 1 - 5, n5 = 10 - 30.

2. The GIP agonist according to claim 1, wherein AA1 is selected as Phe, AA2 is selected as γGlu, AA3 is selected as amino group, n1 = 1 - 5, n2 = 1 - 30, n3 = 0 or = 1 - 5, n4 = 0 or = 1 - 5, n5 = 10 - 30.

3. The GIP agonist according to claim 2, wherein, AA1 is selected as Phe, AA2 is selected as γGlu, AA3 is selected as amino group, n1 = 1, n2 = 5, n3 = 1, n4 = 1, n5 = 18.

4. The GIP agonist according to claim 1, wherein AA1 is selected as αMePhe, AA2 is selected as γGlu, AA3 is selected as amino group, n1 = 1 - 5, n2 = 1 - 30, n3 = 0 or = 1 - 5, n4 = 0 or = 1 - 5, n5 = 10 - 30.

5. The GIP agonist according to claim 4, wherein AA1 is selected as αMePhe, AA2 is selected as γGlu, AA3 is selected as amino group, n1 = 1, n2 = 5, n3 = 1, n4 = 1, n5 = 18.

6. The GIP agonist according to claim 1, wherein AA1 is selected as αMePhe, AA2 is selected as δAad, AA3 is selected as amino group, n1 = 1 - 5, n2 = 1 - 30, n3 = 0 or = 1 - 5, n4 = 0 or = 1 - 5, n5 = 10 - 30.

7. The GIP agonist according to claim 6, characterized in that, AA1 is selected as αMePhe, AA2 is selected as δAad, AA3 is selected as amino group, n1 = 1, n2 = 5, n3 = 1, n4 = 1, n5 = 18.

8. The GIP agonist according to claim 1, wherein AA1 is selected as αMePhe(2F), AA2 is selected as γGlu, AA3 is selected as amino group, n1 = 1 - 5, n2 = 1 - 30, n3 = 0 or = 1 - 5, n4 = 0 or = 1 - 5, n5 = 10 - 30.

9. The GIP agonist according to claim 8, wherein, AA1 is selected as αMePhe(2F), AA2 is selected as γGlu, AA3 is selected as amino group, n1 = 1, n2 = 5, n3 = 1, n4 = 1, n5 = 18.

10. The GIP agonist according to claim 1, characterized in that, AA1 is selected as αMePhe(2F), AA2 is selected as δAad, AA3 is selected as amino group, n1 = 1 - 5, n2 = 1 - 30, n3 = 0 or = 1 - 5, n4 = 0 or = 1 - 5, n5 = 10 - 30.

11. The GIP agonist according to claim 10, wherein AA1 is selected as αMePhe(2F), AA2 is selected as δAad, AA3 is selected as amino group, n1 = 1, n2 = 5, n3 = 1, n4 = 1, n5 = 18.

12. The GIP agonist according to any one of claims 1-11, characterized in that, The triple agonist compound comprises a medicinal salt, chelate or non - covalent complex formed by the compound, and a precursor of the compound, or any mixture of the above forms.

13. Use of a GIP agonist as described in any one of claims 1 - 12 in the preparation of a pharmaceutical composition for treating diseases.

14. The pharmaceutical composition for treating a disease according to claim 13, wherein The pharmaceutical composition for treating diseases is used for at least one of the following diseases, and the diseases include diabetes, obesity, metabolic syndrome, dyslipidemia, atherosclerosis, liver fibrosis diseases, pulmonary fibrosis and other diseases.