Cell and method for detecting activity of GLP-1R / GIPR double-target agonist drug

Abstract

The invention discloses a cell and a method for detecting the activity of a GLP-1R / GIPR double-target agonist drug. The invention relates to a cell for co-expressing GLP-1R, GIPR and CRE-miniP-luciferase reporter genes, and provides a method for detecting the biological activity of a GLP-1R / GIPR double-target agonist drug based on the cell, and the method is simple to operate, short in test period, sensitive in reaction, small in variability, strong in specificity and high in accuracy, can be used for release inspection of products, and can be used for detecting the biological activity of the GLP-1R / GIPR double-target agonist drug. The method has important significance on quality control and clinical application of GLP-1R / GIPR double-target agonist drugs.

Description

Technical Field

[0001] The present invention belongs to the field of biological activity detection of biological drugs. Specifically, the present invention relates to a cell and a method for detecting the activity of a GLP-1R / GIPR dual-target agonist drug. Background Art

[0002] GLP-1R / GIPR dual-target agonists are a new type of drug that achieves multiple biological effects such as blood sugar reduction, weight loss and metabolic improvement by simultaneously activating the glucagon-like peptide-1 receptor (GLP-1R) and the gastric inhibitory peptide receptor (GIPR). This drug design aims to integrate the beneficial effects of multiple gastrointestinal hormones to improve metabolic disorders, especially metabolic diseases such as obesity and diabetes.

[0003] Biological activity can reflect the correctness of the therapeutic protein's high-level structure, the integrity of its biological function, and the batch-to-batch consistency of the product, and is a key quality attribute of GLP-1R / GIPR dual-target agonist drugs. Traditional methods for determining the biological activity of GLP-1R / GIPR dual-target agonist drugs include the use of luminescent beads to detect cAMP and Promega kits to detect residual ATP, but these methods are greatly affected by the environment, have a short half-life, high cost, and poor stability. Therefore, it is of great significance to explore new methods for detecting the activity of GLP-1R / GIPR dual-target agonist drugs. Summary of the invention

[0004] In order to make up for the deficiencies of the prior art, the purpose of the present invention is to provide a cell and method for detecting the activity of GLP-1R / GIPR dual-target agonist drugs.

[0005] In order to achieve the above object, the present invention adopts the following technical solution: A first aspect of the present invention provides a cell line.

[0006] Further, the cell line is named GLP1R / GIPR-luc-293, deposited with the China Culture Collection of Microorganisms, address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, with the deposit number CGMCC No.46309, classified as Human HEK293 cells, and the deposit date is December 25, 2024.

[0007] Furthermore, the cell line co-expresses GLP-1R, GIPR and CRE-miniP-luciferase reporter genes; the CRE-miniP is a fusion gene formed by connecting the CRE gene and the miniP gene.

[0008] Furthermore, the sequence of the CRE gene is shown as SEQ ID NO.1, the sequence of the miniP gene is shown as SEQ ID NO.2, and the sequence of the CRE-miniP gene is shown as SEQ ID NO.3.

[0009] In the present invention, the applicant transfected the CRE-miniP-luciferase reporter gene, the GLP-1 receptor (GLP-1R) expression gene, and the GIP receptor (GIPR) expression gene into HEK293 cells, and obtained 5 stable monoclonal cell lines by pressure screening, monoclonal isolation culture and flow analysis. After functional verification, it was found that the 5 cell lines responded to the GLP-1R / GIPR dual-target agonist tirpotide to varying degrees. Among them, monoclonal clone C9 had the strongest response to tirpotide, GLP-1 and GIP, and the ratios of the upper and lower asymptotes of the curve were 162, 173, and 166, respectively. Therefore, this cell was selected for preservation and named GLP1R / GIPR-luc-293.

[0010] A second aspect of the present invention provides a cell preparation.

[0011] Furthermore, the cell preparation includes the cell line described in the first aspect of the present invention.

[0012] Preferably, the cell preparation further comprises a luciferase reaction substrate.

[0013] Preferably, the cell preparation includes a suspension-type cell preparation, a solution-type cell preparation and a solid dispersion-type cell preparation.

[0014] A third aspect of the present invention provides an evaluation system.

[0015] Furthermore, the evaluation system includes the following components: the cell line described in the first aspect of the present invention, a GLP-1R / GIPR dual-target agonist drug sample and a reference product.

[0016] In the present invention, the term "system" refers to the drug quality control system, which refers to the control of various factors in the drug production process through scientific management methods to ensure that the drug quality meets the predetermined quality standards and requirements.

[0017] In the present invention, the term "sample" refers to a test sample to be identified and determined. In the present invention, GLP-1R / GIPR dual-target agonist drug samples include drug samples produced in industrial production and to be tested for their biological activity.

[0018] A fourth aspect of the present invention provides a product.

[0019] Furthermore, the product includes the cell line described in the first aspect of the present invention or the cell preparation described in the second aspect of the present invention.

[0020] Preferably, the kit further comprises a luciferase reaction substrate and a diluent.

[0021] Preferably, the product is of the type of a kit.

[0022] In the present invention, the components of the test kit can be packaged in the form of an aqueous medium or in a lyophilized form. Suitable containers in the test kit usually include at least a vial, test tube, flask, PET bottle, syringe or other container, in which a component can be placed, and preferably, can be appropriately divided. When there is more than one component in the test kit, the test kit will usually also include a second, third or other additional container, in which additional components are placed separately. However, components of different combinations can be contained in a vial. The test kit of the present invention will usually also include a container for accommodating reactants, sealed for commercial sale. This container can include a plastic container of injection molding or blow molding, in which the required vial can be retained.

[0023] The fifth aspect of the present invention provides a method for constructing the cell line described in the first aspect of the present invention.

[0024] Further, the method comprises: 1) Transfecting effector cells with vectors expressing GLP-1R gene, GIPR gene, and / or CRE-miniP gene; 2) Screening and obtaining monoclonal cell lines that co-express GLP-1R, GIPR and CRE-miniP.

[0025] Furthermore, the CRE-miniP is a fusion gene formed by connecting the CRE gene and the miniP gene.

[0026] Furthermore, the sequence of the CRE gene is shown as SEQ ID NO.1, the sequence of the miniP gene is shown as SEQ ID NO.2, and the sequence of the CRE-miniP gene is shown as SEQ ID NO.3.

[0027] Those skilled in the art can select different methods to synthesize the CRE-miniP gene according to the sequences of the CRE gene and the miniP gene. In a specific embodiment of the present invention, the CRE gene and the miniP gene are handed over to Suzhou Dongling Biotechnology Co., Ltd. for connection using gene total synthesis technology.

[0028] Furthermore, the effector cells are HEK293 cells.

[0029] Furthermore, the expression vector includes a plasmid vector, a phage vector, and a viral vector.

[0030] Preferably, the expression vector is a plasmid vector.

[0031] Furthermore, the transfection method includes physical mediation, chemical mediation, and biological mediation.

[0032] Preferably, the transfection method is biologically mediated.

[0033] More preferably, the transfection method is lentiviral infection.

[0034] In some embodiments, the cell types commonly used for transfection are diverse, including but not limited to the following: 1) Primary cells and their derivatives, which are directly isolated from biological tissues without long-term in vitro culture and maintain high physiological activity and differentiation potential; 2) Tumor cell lines: such as human breast cancer cells (MCF-7), human lung cancer cells (A549), human cervical cancer cells (HeLa), etc. These cell lines have high division rate and stability and are often used for gene function research and drug screening; 3) Cell lines of specific tissue origin: Human embryonic kidney cells (HEK-293): have high transfection efficiency and stability, and are often used in gene cloning and vaccine production; Human embryonic lung cells (HEL): used as recipient cells in some experiments; Human umbilical vein endothelial cells (HUVEC): used to study vascular biology and cardiovascular diseases; 4) Animal cell lines: Baby hamster kidney cells (BHK-21): widely used in virology research and other biomedical research; Mouse embryonic fibroblasts (NIH3T3): have low immunogenicity and high division rate, and are often used for gene knockout and drug screening; Other special cell types: Neuronal cells: used in neurobiological research; Rat hepatocytes: used for drug metabolism and toxicity studies; Rat glioma cells (C6): Highly differentiated and stable, often used in drug screening and tumor research. Those skilled in the art can select different effector cells according to the prior art. In a specific embodiment of the present invention, the effector cell is HEK293.

[0035] In the present invention, the term "reporter gene" is a gene that encodes a protein or enzyme that is easily detected in the background of endogenous proteins. The reporter gene is connected to the target gene or regulatory sequence, and the transcriptional activity or expression level of the gene in the cell is directly reflected by indirectly or directly detecting the signal of the reporter gene encoding product, such as reporter protein, mRNA, enzyme, etc. The characteristics of general reporter genes are non-toxic, non-immunogenic, easy to detect, and have a certain specificity.

[0036] In a specific embodiment of the present invention, the reporter gene used is the LUC gene, i.e., the luciferase reporter gene. Luciferase comes from organisms that can emit light in nature. It is a general term for a class of enzymes that can catalyze the oxidation of luciferin or fatty aldehydes to produce light. Depending on the source organism, luciferase can be divided into firefly luciferase (FL) and bacterial luciferase (BL). At present, the luciferase gene from the North American firefly is the most widely used. This gene can encode a luciferase protein of 550 amino acids. The FL gene comes from fireflies and is found in Mg. 2+ , ATP, O2, catalyzes the oxidative decarboxylation of D-luciferin to produce activated oxyluciferin, and emits photons to produce 550-580nm fluorescence. Luciferase reporter gene detection is an important tool in the field of modern molecular biology research to analyze the interaction between potential cis elements (such as promoters, enhancers and silencers, etc.) and trans-acting factors in the flanking regions of structural genes. The luciferase reporter gene system is a reporting system that uses luciferin as a substrate to detect the activity of firefly luciferase. Luciferase can catalyze the oxidation of luciferin to oxyluciferin. During the oxidation process of luciferin, bioluminescence will be emitted, which is then measured by a chemiluminometer or liquid scintillation meter.

[0037] In the present invention, the term "plasmid" is a naked double-stranded circular DNA molecule independent of the DNA in the cell nucleus, which has the ability to replicate autonomously and can be stably inherited in the cell. Plasmids are mainly found in bacteria, but can also be found in eukaryotes. Specific DNA sequences are introduced into the plasmid, which encode a certain or certain specific proteins, thereby giving the host cell new properties or functions. For example, the plasmid can carry a target gene for gene therapy, or a coding sequence for protein production, or give the cell a new function through transfection. In the present invention, those skilled in the art can select different plasmids to introduce the GLP-1R gene, GIPR gene and CRE-miniP-luciferase reporter gene into the cell. In a specific embodiment of the present invention, the vector for introducing the GLP-1R gene and GIPR gene into the cell is the pLU.cmv.neo plasmid, and the vector for introducing the CRE-miniP-luciferase reporter gene into the cell is the pLU.CMV.Luc.PGK.PURO plasmid.

[0038] In the present invention, the term "transfection" refers to the process of introducing exogenous DNA (such as plasmid) into eukaryotic cells to express exogenous genes in cells. Plasmid transfection is an important technology in genetic engineering and is widely used in gene function research, protein production and other fields.

[0039] Methods of cell transfection include physical mediation, chemical mediation, and biological mediation. The physical mediation includes electroporation, microinjection, and gene gun; the chemical mediation includes calcium phosphate coprecipitation, cationic polymer, and cationic liposome; the biological mediation is mainly viral mediation, such as lentivirus, adenovirus, etc. In a specific embodiment of the present invention, the transfection method is lentivirus transfection. Furthermore, the screening includes the following steps: pressure screening, monoclonal isolation and culture, and flow cytometry analysis.

[0040] Preferably, the pressure screening is performed by adding antibiotics.

[0041] Furthermore, the antibiotics are bleomycin, puromycin and G418.

[0042] More preferably, the concentrations of bleomycin, puromycin and G418 are 200 μg·mL -1 , 3μg·mL -1 and 1000 μg mL -1 .

[0043] In the present invention, the term "pressure screening" refers to the screening and retention of successfully transfected cells by a specific selection pressure. Generally, pressure screening methods include antibiotic screening and gene expression product screening. In a specific embodiment of the present invention, the pressure screening is performed by adding antibiotics.

[0044] In cell screening, antibiotics are often used as a resistance screening tool to distinguish successfully transfected cells from untransfected cells. These antibiotics inhibit the growth of untransfected cells or kill them, thereby allowing cells that are successfully transfected and express antibiotic resistance genes to survive.

[0045] Further, the antibiotics include but are not limited to: a penicillin-streptomycin mixture, G-418 (Geneticin), Blasticidin, Hygromycin B, Puromycin, 6-thioguanine, Ampicillin, and Zeocin. In a specific embodiment of the present invention, the antibiotics are zeocin, Puromycin, and G418.

[0046] The sixth aspect of the present invention provides a method for detecting the biological activity of a GLP-1R / GIPR dual-target agonist drug.

[0047] Furthermore, the method comprises: adding a GLP-1R / GIPR dual-target agonist drug to the cell line described in the first aspect of the present invention for incubation, adding an enzyme reaction substrate after incubation, and determining the biological activity of the GLP-1R / GIPR dual-target agonist drug based on the measured reporter gene signal value.

[0048] Furthermore, when the GLP-1R / GIPR dual-target agonist drug is incubated with the cell line described in the first aspect of the present invention, its initial concentration is 25nmol·mL -1 ~100nmol·mL -1 .

[0049] Preferably, the initial concentration of the GLP-1R / GIPR dual-target agonist drug is 25nmol·mL -1 .

[0050] Furthermore, the dilution multiple of the GLP-1R / GIPR dual-target agonist drug is 4 to 6 times.

[0051] More preferably, the dilution multiple of the GLP-1R / GIPR dual-target agonist drug is 4 times.

[0052] Furthermore, the GLP-1R / GIPR dual-target agonist drugs include telpotide, BGM0504 and HS-20094.

[0053] Preferably, the GLP-1R / GIPR dual-target agonist drug is telpotide.

[0054] In the present invention, the applicant optimized the starting concentration and dilution multiple of the GLP-1R / GIPR dual-target agonist drug telportin and found that the optimal starting concentration was 25 nmol·mL -1 The best dilution factor is 4 times.

[0055] In the present invention, after measuring the reporter gene signal value, a four-parameter curve is fitted to determine the biological activity of the GLP-1R / GIPR dual-target agonist drug. The term four-parameter curve fitting is a mathematical method used to describe a set of data by fitting a curve and obtain a functional equation that can describe the data relationship in order to predict future data. The four-parameter curve is usually defined by four parameters, which have practical significance in the biological context. For example, in detecting the biological activity of a drug, these four parameters may represent: the lower limit of absorbance (D), the upper limit of absorbance (A), that is, the minimum and maximum values ​​of absorbance; the half-response concentration (C), that is, the value when the drug concentration reaches half the maximum response; and the slope of the curve (B). Through these parameters, the experimental data can be understood and analyzed more accurately, thereby providing deeper insights into biological research. A seventh aspect of the present invention provides applications of any of the following aspects.

[0056] Furthermore, the application includes: 1) Use of the cell line described in the first aspect of the present invention or the cell preparation described in the second aspect of the present invention in the preparation of a product for detecting the biological activity of a GLP-1R / GIPR dual-target agonist drug; 2) Application of the cell line described in the first aspect of the present invention, the cell preparation described in the second aspect of the present invention, or the evaluation system described in the third aspect of the present invention in the quality control of GLP-1R / GIPR dual-target agonist drugs; 3) Application of the evaluation system described in the third aspect of the present invention in evaluating the biological activity of GLP-1R / GIPR dual-target agonist drugs.

[0057] Advantages and beneficial effects of the present invention: The present invention provides a cell and method for detecting the activity of a GLP-1R / GIPR dual-target agonist drug. The principle of the method is that after the GLP-1 receptor and the GIP receptor of the CRE-miniP-luc-GLP-1R-GIPR HEK293 monoclonal cell line bind to telpotide, a series of signal transductions can activate the expression of the reporter gene luciferase, and the telpotide activity is quantified by detecting the change in luciferase expression after telpotide stimulation. The method has the advantages of simple operation, short test cycle, sensitive reaction, small variability, strong specificity and high accuracy, and can be used for product release inspection, which is of great significance for the quality control and clinical application of telpotide. BRIEF DESCRIPTION OF THE DRAWINGS

[0058] Figure 1 Shown are the results confirming that the CRE-miniP-luc-GLP-1R-GIPR HEK293 monoclonal cell line expresses GLP1-R; Figure 2 Shown are the results confirming that the CRE-miniP-luc-GLP-1R-GIPR HEK293 monoclonal cell line expresses GIPR; Figure 3 Shows the response of the CRE-miniP-luc-GLP-1R-GIPR HEK293 monoclonal cell line to tepoxetine, GLP-1, and GIP; Figure 4 Figure showing the results of the optimization of the starting concentration of tepoxetine; Figure 5 Graph showing the results of optimization of the dilution factor of tepoxetine. DETAILED DESCRIPTION

[0059] The present invention is further described below in conjunction with specific embodiments, which are only used to explain the present invention and cannot be understood as limiting the present invention. Those skilled in the art can understand that: various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and purpose of the present invention, and the scope of the present invention is defined by the claims and their equivalents.

[0060] The experimental methods in the following examples without specifying specific conditions are usually carried out according to conventional conditions or conditions recommended by the manufacturer.

[0061] Example 1 Cell construction and positive cell screening 1. Experimental Methods The GLP-1R gene was constructed into the pLU.cmv.neo vector through the BamHI and SalI cloning sites, the GIPR gene was constructed into the pLU.cmv.zeo vector through the BamHI and SalI cloning sites, and the CRE-miniP gene was constructed into the pLU.CMV.Luc.PGK.PURO vector through the ClaI and XbaⅠ cloning sites. The plasmids were then amplified and purified, and the lentivirus was packaged with a virus packaging kit to infect HEK293T cells. Then, 200 μg mL -1 zeocin, 3 μg mL -1 puromycin, 1000 μg mL -1 The cells were screened with G418 to obtain stable transfected cells. Finally, the expression of GLP-1R and GIPR on the cell surface was analyzed by flow cytometry to determine the positive monoclonal cells.

[0062]

[0063] 2. Experimental Results Five monoclonal cell lines were obtained by limiting dilution method. The GLP-1R positive ratios of the five monoclonal cell lines B6, B9, C9, E10 and G8 were 99.2%, 99.3%, 96.0%, 91.7% and 96.9%, respectively; the GIPR positive ratios were 96.3%, 98.8%, 98.4%, 98.1% and 97.4%, respectively. Figure 1 and Figure 2 ). The five cell lines were cultured at 4.0×10 4 The cells were plated at a density of 100 cells / well and the biological activities of the same sample of telportin, GLP-1, and GIP were determined. Figure 3 As shown in the figure, all five cell lines had a dose-effect response to tebuconazole stimulation, and the level of luciferase expressed by cells increased with the increase of drug concentration. The ratio of the upper and lower asymptotes of the S curve fitted by the four-parameter model of the five cell lines was greater than 40. Among them, the C9 monoclonal cell had the strongest response to tebuconazole, GLP-1 and GIP, and the ratio of the upper and lower asymptotes of the curve were 162, 173 and 166, respectively. Therefore, the C9 monoclonal cell line was selected for subsequent experiments.

[0064] Example 2 Method parameter optimization 1. Experimental Methods Dilute tepol to a starting concentration of 100 nmol mL -1 , 50nmol·mL -1 , 25nmol·mL -1 After that, 6-fold gradient dilutions were performed, with a total of 10 concentration points, to verify the response of C9 cell line to tepoxetine. The optimal starting concentration was selected to further optimize the dilution multiple.

[0065] 2. Experimental Results like Figure 4 As shown in the figure, the three starting concentrations can all reach the platform on the curve. When the starting concentration is 25nmol·mL -1 When the curve is 25 nmol·mL, a clear upper platform is formed, but there are only two points in the linear part, so 25 nmol·mL is selected. -1 The dilution factor was further optimized for the starting concentration. The starting concentration of 25 nmol·mL-1 was diluted 4-fold, 5-fold, and 6-fold, respectively, with 10 concentration points. When the dilution factor was 4-fold, there were 3 points on the upper and lower platforms of the curve, and the linear part had the most points ( Figure 5 ), so the optimal starting concentration of the method was determined to be 25nmol·mL -1 The best dilution factor is 4 times.

[0066] The description of the above embodiments is only used to understand the method and core idea of ​​the present invention. It should be pointed out that, for those skilled in the art, several improvements and modifications can be made to the present invention without departing from the principle of the present invention, and these improvements and modifications will also fall within the scope of protection of the claims of the present invention.

Claims

1. A cell line, characterized in that The cell strain is named GLP1R / GIPR-luc-293, deposited in the China Microbiological Culture Collection, address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, with the deposit number CGMCC No. 46309, classified as Human HEK293 cells, and the deposit date is December 25, 2024.

2. The cell line according to claim 1, characterized in that The cell line co-expresses GLP-1R, GIPR and CRE-miniP-luciferase reporter genes; the CRE-miniP is a fusion gene formed by connecting the CRE gene and the miniP gene; Preferably, the sequence of the CRE gene is shown as SEQ ID NO.1, the sequence of the miniP gene is shown as SEQ ID NO.2, and the sequence of the CRE-miniP gene is shown as SEQ ID NO.

3.

3. A cell preparation, characterized in that: The cell preparation comprises the cell line according to claim 1; Preferably, the cell preparation further comprises a luciferase reaction substrate; Preferably, the cell preparation includes a suspension-type cell preparation, a solution-type cell preparation and a solid dispersion-type cell preparation.

4. An evaluation system, characterized in that: The evaluation system comprises the following components: the cell line according to claim 1 or 2, a GLP-1R / GIPR dual-target agonist drug sample and a reference product.

5. A product, characterized in that The product includes the cell line according to claim 1 or 2 or the cell preparation according to claim 3; Preferably, the kit further comprises a luciferase reaction substrate and a diluent; Preferably, the product is of the type of a kit.

6. The method for constructing the cell line according to claim 1 or 2, characterized in that: The method comprises: 1) Transfecting effector cells with vectors expressing GLP-1R gene, GIPR gene, and / or CRE-miniP gene; 2) Screening for monoclonal cell lines that co-express GLP-1R, GIPR, and CRE-miniP; Preferably, the CRE-miniP is a fusion gene formed by connecting the CRE gene and the miniP gene; Preferably, the sequence of the CRE gene is shown as SEQ ID NO.1, the sequence of the miniP gene is shown as SEQ ID NO.2, and the sequence of the CRE-miniP gene is shown as SEQ ID NO.

3.

7. The method according to claim 6, characterized in that The effector cells are HEK293 cells; Preferably, the expression vector includes a plasmid vector, a phage vector, or a viral vector; Preferably, the expression vector is a plasmid vector; Preferably, the transfection method includes physical mediation, chemical mediation, and biological mediation; Preferably, the transfection method is biologically mediated; More preferably, the transfection method is lentiviral infection; Preferably, the screening comprises the following steps: pressure screening, monoclonal isolation and culture, and flow analysis; Preferably, the pressure screening is performed by adding antibiotics; Preferably, the antibiotics are bleomycin, puromycin and G418; More preferably, the concentrations of bleomycin, puromycin and G418 are 200 μg·mL-1, 3 μg·mL-1 and 1000 μg·mL-1, respectively.

8. A method for detecting the biological activity of a GLP-1R / GIPR dual-target agonist drug, characterized in that: The method comprises: adding a GLP-1R / GIPR dual-target agonist drug to the cell line of claim 1 or 2 for incubation, adding an enzyme reaction substrate after incubation, and determining the biological activity of the GLP-1R / GIPR dual-target agonist drug according to the measured reporter gene signal value.

9. The method according to claim 8, characterized in that When the GLP-1R / GIPR dual-target agonist drug is incubated with the cell line according to claim 1 or 2, its initial concentration is 25nmol·mL-1~100nmol·mL-1; Preferably, the initial concentration of the GLP-1R / GIPR dual-target agonist drug is 25nmol·mL-1; Preferably, the dilution multiple of the GLP-1R / GIPR dual-target agonist drug is 4 to 6 times; More preferably, the dilution multiple of the GLP-1R / GIPR dual-target agonist drug is 4 times; Preferably, the GLP-1R / GIPR dual-target agonist drugs include telpotide, BGM0504 and HS-20094; More preferably, the GLP-1R / GIPR dual-target agonist drug is telpotide.

10. The application of any one of the following aspects, characterized in that: The applications include: 1) Use of the cell line according to claim 1 or 2 or the cell preparation according to claim 3 in the preparation of a product for detecting the biological activity of a GLP-1R / GIPR dual-target agonist drug; 2) Application of the cell line according to claim 1 or 2, the cell preparation according to claim 3, or the evaluation system according to claim 4 in the quality control of GLP-1R / GIPR dual-target agonist drugs; 3) Application of the evaluation system described in claim 4 in evaluating the biological activity of GLP-1R / GIPR dual-target agonist drugs.

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