Label-free cell screening model of gpr183 receptor and its application in preparation of drugs

Abstract

This invention relates to a drug screening method for diseases closely related to label-free G protein-coupled receptors (GPCRs), specifically to a label-free cell screening model for the GPR183 receptor and the application of drugs prepared from it. The label-free cell screening model is a stable cell line CHO-K1-GPR183 that highly expresses the GPR183 receptor. The GPR183 receptor agonist compound N-demethylnuciferine was obtained through screening using the GPR183 cell screening model established in this invention. Current research indicates that the GPR183 receptor is closely related to the occurrence and development of neuroinflammatory / autoimmune diseases such as multiple sclerosis, colitis, type 1 diabetes, and arthritis, as well as cancer and metabolic diseases, thus providing potential lead compounds for drug development in these diseases.

Description

Technical Field

[0001] This invention relates to a drug screening method for diseases closely related to label-free G protein-coupled receptors (GPCRs), specifically to a label-free cell screening model for the GPR183 receptor and the application of drugs prepared from it. Background Technology

[0002] G protein-coupled receptors (GPCRs) are seven-transmembrane receptors and are among the most important targets in drug development. Currently, more than 30% of marketed drugs target GPCRs [ASHauser, M. M. Attwood, M. Rask-Andersen, et al. Nat. Rev. Drug Discov. 2017, 16, 829]. GPR183 is expressed on B cells, T cells, dendritic cells (DCs), macrophages, and innate lymphocytes. Studies have found that oxosterol is a ligand for GPR183, and its interaction with GPR183 can mediate the migration and localization of immune cells (including innate lymphocytes) [B. Misselwitz, A. Wyss, T. Raselli, et al. Br J Pharmacol. 2021, 178, 3140]. Current research indicates that the GPR183 receptor is associated with neuroinflammatory / autoimmune diseases, such as multiple sclerosis, colitis, type 1 diabetes, and arthritis. The GPR183 receptor and its endogenous ligands also play important roles in various diseases, including B-cell malignancies and metabolic disorders [K. Braden, M. Campolo, Y. Li, et al. J Pharmacol Exp Ther. 2022, 383, 172; VMSKjaer, L. Ieremias, V. Daugvilaite. ChemMedChem. 2021, 16, 2623]. Therefore, research on the ligand regulation and signal transduction mechanisms of GPR183 is of significant guiding importance for its biological function and drug development. Thus, the search for and development of highly active GPR183 ligands is essential.

[0003] Currently, high-throughput screening methods mainly include traditional radioligand-receptor binding assays, GTPγS binding assays, cyclic adenosine monophosphate (cAMP) assays, calcium flux assays, reporter gene assays, receptor endocytosis assays, and β-arrestin recruitment assays. However, these methods all have certain limitations. Most GPCR assays target only one signaling pathway and cannot simultaneously detect the activation of multiple signaling pathways. Furthermore, they require the addition of fluorescent labels or additional indicators, making the operation cumbersome, and the indicators can also have some impact on cells [W. Thomsen, J. Frazer, D. Unett. Curr Opin Biotechnol. 2005, 16, 655; L.M. Mayr, D. Bojanic. Curr Opin Pharmacol. 2009, 9, 580]. Novel label-free integrative pharmacology technology utilizes label-free resonant waveguide grating (RWG) biosensors to transform the dynamic mass redistribution of intracellular components induced by drugs into a holistic, dynamic wavelength shift response signal, known as dynamic mass reset (DMR) signal. This technology is characterized by being non-invasive, having high spatiotemporal resolution, high sensitivity, high throughput, suitability for target-pathway integration studies, ease of operation, and short experimental cycles. The detection process requires no labeling or additional indicators, providing a more realistic reflection of drug effects at the whole-cell level [M. Morse, H. Sun, E. Tran, et al. BMCPharmacol Toxicol. 2013, 14, 17]. Currently, there is no method for constructing a screening model for the membrane receptor GPR183 using this technology. Therefore, this invention employs label-free integrative pharmacology technology to construct a label-free high-throughput screening model for the GPR183 receptor, which can significantly improve the efficiency of GPR183 ligand screening. This is of great significance for elucidating the pharmacological and physiological functions of GPR183 and provides a model system for drug screening in diseases closely related to the GPR183 receptor.

[0004] In 1993, the Epstein-Barr virus-induced orphan receptor GPR183 was discovered in the Burkitt lymphoma cell line. Its ligand remained unknown until two landmark publications in 2011 identified dehydroxylated oxysterols as ligands for GPR183, with 7α,25-dihydroxycholesterol being the most potent [B. Misselwitz, A. Wyss, T. Raselli, et al. Br J Pharmacol. 2021, 178, 3140]. Based on the work identifying and synthesizing GPR183 ligands, a few small molecules were derived. Prior to GPR183 deorphanization, a small-molecule inverse agonist of GPR183 named GSK682753 was identified. In 2014, a library containing approximately 100K compounds was screened, revealing the GPR183 agonist NIBR51. Based on NIBR51, the same compound library was re-screened to identify the antagonist NIBR127. Chemical optimization yielded the more effective GPR183 antagonist NIBR189 [VMSKjaer, L. Ieremias, V. Daugvilaite, et al. ChemMedChem. 2021, 16, 2623]. Currently, N-demethylnuciferine has not been reported as a ligand for GPR183. In this invention, the GPR183 agonist N-demethylnuciferine was obtained through screening using a label-free GPR183 cell model. This provides a prospective compound for drug development for GPR183-related diseases such as multiple sclerosis, colitis, type 1 diabetes, and neuroinflammatory / autoimmune diseases, cancer, and metabolic diseases. Summary of the Invention

[0005] The purpose of this invention is to provide a label-free cell screening model for the GPR183 receptor and its application in drug preparation.

[0006] To achieve the above objectives, the following technical solution is provided using novel label-free cell integration pharmacology technology:

[0007] A label-free cell selection model for the GPR183 receptor, wherein the label-free cell selection model is a stable cell line CHO-K1-GPR183 that highly expresses the GPR183 receptor.

[0008] The stable cell line CHO-K1-GPR183 was obtained by transfecting CHO-K1 cells with a plasmid containing GPR183.

[0009] Application of a label-free cell screening model for the GPR183 receptor, wherein the label-free cell screening model is used in screening drugs for diseases related to the GPR183 receptor.

[0010] The label-free cell screening model is used in screening drugs for the prevention and / or treatment of diseases by activating or antagonizing the GPR183 receptor.

[0011] The disease is one or more of the following: multiple sclerosis, colitis, type 1 diabetes, neuroinflammatory / autoimmune diseases, cancer, and metabolic diseases.

[0012] A method for screening drugs for diseases closely related to the GPR183 receptor is based on label-free cell integrative pharmacology. Utilizing the stable GPR183-expressing cell line CHO-K1-GPR183 as described in claim 1, the method determines the agonist or antagonist activity of the test sample based on the similarity and specificity of the DMR signal spectrum of the test sample with the characteristic DMR signal spectra of known standard agonists and / or antagonists, thereby screening for drugs closely related to diseases of the GPR183 receptor.

[0013] The label-free cell integrated pharmacology technology utilizes a resonant waveguide grating (RWG) biosensor to convert the dynamic redistribution of intracellular components caused by drugs into a holistic, dynamic wavelength shift response signal. This signal is the response value (pm) of wavelength change, which is realized through an Epic optical biosensor 384 microplate.

[0014] The known standard agonist is 7α,25-dihydroxycholesterol; the known standard antagonist is NIBR189.

[0015] The cell screening model for the GPR183 receptor involved seeding CHO-K1-GPR183 cells into 384-well microplates with optical biosensing capabilities at a cell-compatible density of 1.5 × 10⁻⁶ cells / well. 4 Cells per well, with a cell culture medium volume of 40 μL per well, and a cell culture time of 18–24 h after seeding.

[0016] The screening protocol for agonists is as follows:

[0017] (1) Samples of different concentrations (80 μM, 40 μM, 20 μM, 10 μM, 5 μM, 2.5 μM, 1.25 μM) or different concentrations of GPR183 receptor agonist 7α,25-dihydroxycholesterol (10 μM, 5 μM, 2.5 μM, 1.25 μM, 0.625 μM, 0.313 μM, 0.156 μM, 0.078 μM) were dissolved in HBSS buffer. (0.039 μM, 0.020 μM, 0.010 μM, 0.005 μM, 0.002 μM, 0.001 μM) were added to 384-well microplates seeded with CHO-K1-GPR183 cells, respectively. The plates were placed on an Epic instrument to monitor the DMR characteristic signal spectrum in real time for 60 min. If the DMR characteristic spectrum of the sample and the GPR183 receptor agonist 7α,25-dihydroxycholesterol have contour similarity;

[0018] (2) The GPR183 receptor antagonist NIBR189 (concentrations of 100 nM, 25 nM, 6.25 nM, 1.56 nM, 0.39 nM, 0.10 nM, and 0.02 nM) dissolved in HBSS buffer salt was added to a 384-well microplate seeded with CHO-K1-GPR183 cells for 60 min of pretreatment. Then, the sample (sample concentration: the concentration at which the sample in step (1) caused 80%-100% DMR characteristic signal intensity) was added to the wells of the cell plate containing the antagonist NIBR189, and its DMR characteristic signal was detected. If the intensity of this DMR characteristic signal was lower than that in step (1), the sample was determined to be an agonist of the GPR183 receptor.

[0019] The screening protocol for antagonists is as follows:

[0020] (1) Different concentrations of samples (80 μM, 40 μM, 20 μM, 10 μM, 5 μM, 2.5 μM, 1.25 μM) dissolved in HBSS buffer or different concentrations of GPR183 receptor agonist 7α,25-dihydroxycholesterol (10 μM, 5 μM, 2.5 μM, 1.25 μM, 0.625 μM, 0.313 μM, 0.156 μM, 0.078 μM, 0.039 μM, 0.020 μM, 0.010 μM, 0.005 μM, 0.002 μM, 0.001 μM) were added to 384-well microplates seeded with CHO-K1-GPR183 cells and placed on an Epic instrument to monitor the DMR characteristic signal spectrum in real time for 60 min.

[0021] (2) If the sample in step (1) does not induce a DMR signal spectrum, add agonist 7α,25-dihydroxycholesterol (concentration of agonist 7α,25-dihydroxycholesterol: the concentration of 7α,25-dihydroxycholesterol in step (1) that induces 80%-100% DMR characteristic signal intensity) to the cell plate in step (1) and detect the DMR characteristic signal spectrum; if this DMR characteristic signal is weaker than the signal of 7α,25-dihydroxycholesterol in step (1), it can be determined that the sample is an antagonist of GPR183 receptor.

[0022] The rising phase lasts 1–30 minutes, and the plateau phase lasts 30–90 minutes.

[0023] The screened GPR183 receptor agonists were one or more of the compounds N-demethylnuciferine or pharmaceutically acceptable salts thereof, and the compounds had the following structural formulas:

[0024]

[0025] The use of a compound in the preparation of a medicament for the prevention and / or treatment of diseases by activating GPR183 receptors; the use of the following structural compound in the preparation of a medicament for the prevention and / or treatment of diseases by activating GPR183 receptors. (N-Demethyl lotus leaf alkaloid).

[0026] The GPR183 receptor agonist is one or more active ingredients of the compound N-demethylnuciferine and its pharmaceutically acceptable salts, and may also contain pharmaceutically acceptable carriers or excipients, such as starch, sodium chloride, microcrystalline cellulose, sorbic acid, and / or mannitol. The composition can be administered via, but is not limited to, intravenous injection, oral administration, intramuscular injection, subcutaneous injection, or local injection. Its dosage form can be, but is not limited to, injection solutions, lyophilized powder for injection, injection microspheres, liposomes, tablets, capsules, aqueous solutions, powders, pastes, sprays, granules, soft capsules, pellets, gels, patches, and ointments, with injection solutions, lyophilized powders, tablets, and capsules being preferred.

[0027] The use of the compound N-demethylnuciferine described in this invention in the preparation of drugs for the prevention and / or treatment of neuroinflammatory / autoimmune diseases, cancer, and metabolic diseases such as multiple sclerosis, colitis, type 1 diabetes, and arthritis, wherein the compound N-demethylnuciferine and one or more of the corresponding pharmaceutically acceptable salts are included.

[0028] The drug is an active ingredient consisting of one or more of the compound N-demethylnuciferine and its pharmaceutically acceptable salts, and may also contain a pharmaceutically acceptable carrier or excipient.

[0029] The advantages and beneficial effects of this invention are as follows:

[0030] The label-free GPR183 cell model established in this invention can be used for high-throughput screening of commercially available synthetic and natural small molecule libraries or natural product extracts / mixtures to obtain highly active ligands of the GPR183 receptor.

[0031] In this invention, the GPR183 agonist N-demethylnuciferine was obtained by screening a label-free GPR183 cell model. The GPR183 receptor plays an important role in neuroinflammatory / autoimmune diseases such as multiple sclerosis, colitis, type 1 diabetes and arthritis, as well as cancer and metabolic diseases. Based on the correlation between the target and the disease, the clinical application scope of this compound can be broadened. Attached Figure Description

[0032] Figure 1 Concentration-response dependence curves of different concentrations of the GPR183 receptor agonist 7α,25-dihydroxycholesterol in CHO-K1-GPR183 cells and concentration-response dependence curves corresponding to the DMR signal spectra of fixed concentrations of 7α,25-dihydroxycholesterol after treating CHO-K1-GPR183 cells for 60 min.

[0033] Figure 2 Concentration-response dependent curves of DMR signal spectra corresponding to fixed concentrations of 7α,25-dihydroxycholesterol after CHO-K1-GPR183 cells were pretreated with different concentrations of the GPR183 receptor antagonist NIBR189 for 60 min.

[0034] Figure 3 The activating and desensitizing signal response values ​​of compound N-demethylnuciferine in CHO-K1-GPR183 cells. Detailed Implementation

[0035] The present invention will now be further illustrated with examples. These examples are merely illustrative and not intended to limit the scope of the invention.

[0036] The novel label-free cell-integrated pharmacology technology used in this model is based on label-free resonant waveguide grating (RWG) biosensors. It transforms the dynamic mass redistribution of intracellular components caused by drugs into a holistic, dynamic wavelength shift response signal, called dynamic mass reset (DMR) signal. It features non-invasiveness, high spatiotemporal resolution, high sensitivity, high throughput, target-pathway integration research, simple operation, and short experimental cycle. The detection process does not require labeling or the addition of additional indicators, and more realistically reflects the effect of drugs at the whole level of living cells.

[0037] In the following examples, the CHO-K1 ovarian cells from the Chinese hamster were obtained from the Cell Bank of the Chinese Academy of Sciences Type Culture Collection Committee. An inverted microscope was purchased from OLYMPUS. 7α,25-dihydroxycholesterol (catalog number: HY-113962) and NIBR189 (catalog number: HY-12336) were purchased from Guangzhou Weijia Technology Co., Ltd. F12K culture medium (catalog number: C11765500BT) was purchased from Gibco. Lipofectamine was used. TM 3000 transfection reagent (catalog number: L3000015) was purchased from Gibco. Antibiotic G418 (catalog number: G4180-5g) was purchased from Tianjin Bai'ao Hengkang Biotechnology Co., Ltd. Balanced salt solutions HBSS (catalog number: 14065-056) and HEPES (catalog number: 15630-080) were purchased from Gibco. Cell culture plates were Epic optical biosensor 384-well microplates, purchased from Corning. The detection platform was Corning's third-generation... The imaging system, purchased from Corning, detects the wavelength shift caused by dynamic mass resetting (DMR) of cells.

[0038] Example 1: Construction of a stable CHO-K1-GPR183 cell line

[0039] Specific transfection methods:

[0040] 1) Transfect CHO-K1 cells with the GPR183 plasmid (8 μg, 1 μg / μL): transfection reagent (24 μL) = 1:3; the GPR183 plasmid was pIRES2-mCherry-GPR183. A cell line CHO-K1-5-GPR183 expressing the GPR183 receptor was obtained.

[0041] 2) Eight hours after transfection, replace with 10 mL of fresh F12 complete culture medium (10% FBS).

[0042] 3) 24 h after transfection, replace with 12 mL of fresh F12 complete medium containing 600 μg / μL G418. 4) Culture for drug screening for 7-10 days, replacing with 12 mL of fresh F12 complete medium containing 600 μg / μL G418 every 2-3 days to obtain a stable cell line CHO-K1-GPR183 (DOI:10.1038 / nature10280) that highly expresses the GPR183 receptor.

[0043] Example 2: Characterization of the GPR183 receptor agonist 7α,25-dihydroxycholesterol in CHO-K1-GPR183 cells

[0044] The CHO-K1-GPR183 cells obtained in the logarithmic growth phase according to the above examples were seeded in cell-compatible 384-well microplates using F12K culture medium. The seeding volume per well was 40 μL, and the seeding density was 1.5 × 10⁻⁶. 4 Incubate the seeded cells in a cell culture incubator for 20–24 hours until cell confluence reaches approximately 95%, then perform cell viability testing. Replace the cell culture medium in the microplate with Hank's balanced salt solution (HBSS, containing 20 mM HEPES), adding 30 μL to each well. After adding the solution, place the plate in an incubator. After equilibration on the imaging system for 120 min, the baseline was rescanned for 2 min. Different concentrations of the GPR183 receptor agonist 7α,25-dihydroxycholesterol (concentrations of 10 μM, 5 μM, 2.5 μM, 1.25 μM, 0.625 μM, 0.313 μM, 0.156 μM, 0.078 μM, 0.039 μM, 0.020 μM, 0.010 μM, 0.005 μM, 0.002 μM, and 0.001 μM) were added to the microplate. 10 μL was added to each well, with three replicates. The plate was placed on an Epic instrument for real-time DMR signal monitoring for 60 min, followed by a 2-min baseline rescan. Then, 10 μL of 7α,25-dihydroxycholesterol (equivalent to 1 μM 7α,25-dihydroxycholesterol according to the curve) was added to each well, with three replicates. The plate was placed on an Epic instrument for real-time DMR signal monitoring for 60 min. Results are shown below. Figure 1 Experimental results showed that 7α,25-dihydroxycholesterol activated the GPR183 receptor to produce a concentration-dependent DMR signal response. The concentration-response curves were monophasic "S"-shaped and all reached saturation, with the highest DMR response value reaching approximately 150 pm. Its EC50... 50 The value was 0.011 ± 0.002 μM; 7α,25-dihydroxycholesterol desensitized GPR183 receptors in a concentration-dependent manner, with the concentration response curves exhibiting a monophasic "S" shape and all reaching saturation responses. Its IC50 value was... 50 The value was 0.103 ± 0.004 μM. These results indicate that the GPR183 receptor (CHO-K1-GPR183 cells) is functionally expressed and can be used for agonist screening and characterization.

[0045] Example 3: Characterization of the GPR183 receptor antagonist NIBR189 in CHO-K1-GPR183 cells

[0046] CHO-K1-GPR183 cells in logarithmic growth phase were seeded into cell-compatible 384-well microplates using F12K culture medium. The seeding volume per well was 40 μL, and the seeding density was 1.5 × 10⁻⁶.4 Incubate the seeded cells in a cell culture incubator for 20–24 hours until cell confluence reaches approximately 95%, then perform cell viability testing. Replace the cell culture medium in the microplate with Hank's balanced salt solution (HBSS, containing 20 mM HEPES), adding 30 μL to each well. After adding the solution, place the plate in an incubator. Equilibrate on the imaging system for 120 min. Different concentrations of the GPR183 antagonist NIBR189 (500 nM, 166.6667 nM, 55.5556 nM, 18.5185 nM, 6.1728 nM, 2.0576 nM, 0.6859 nM, 0.2286 nM, 0.0762 nM, 0.0254 nM, 0.0085 nM, 0.0028 nM, 0.0009 nM, 0.0003 nM) were added to the micro-... Cells were pretreated in microplates for 60 min, with 10 μL added to each well (3 replicates). The baseline was scanned again for 2 min. Then, 1 μM 7α,25-dihydroxycholesterol (the concentration at which 7α,25-dihydroxycholesterol induces 80%-100% of the characteristic DMR signal intensity) was added to each well (10 μL), with 3 replicates. The microplates were then placed on an Epic instrument for real-time DMR signal monitoring for 60 min. Results are shown below. Figure 2 Experimental results showed that NIBR189 antagonized the GPR183 receptor in a concentration-dependent manner, with the concentration-response curves exhibiting a monophasic "S" shape and all reaching saturation responses. Its IC50 value was [missing information]. 50 The value is 1.263 ± 0.0001 nM. These results indicate that the model can be used for the screening and characterization of antagonists.

[0047] Example 4: Evaluation of the activity of compounds N-demethylnuciferine, pseudopunicine, and chithamol in activating the GPR183 receptor.

[0048] CHO-K1-GPR183 cells in logarithmic growth phase were seeded into cell-compatible 384-well microplates using F12K culture medium. The seeding volume per well was 40 μL, and the seeding density was 1.5 × 10⁻⁶. 4 Incubate the seeded cells in a cell culture incubator for 20–24 hours until cell confluence reaches approximately 95%, then perform cell viability testing. Replace the cell culture medium in the microplate with Hank's balanced salt solution (HBSS, containing 20 mM HEPES), adding 30 μL to each well. After adding the solution, place the plate in an incubator. Cells were equilibrated on the imaging instrument for 120 min. Compounds N-demethylnuciferine / pseudococine / chiploalol (all at 40 μM), the GPR183 agonist 7α,25-dihydroxycholesterol (1 μM), and the control group (HBSS buffer containing 0.1% dimethyl sulfoxide) were added to microplates for 60 min pretreatment, in quadruplicate. The baseline was rescanned for 2 min. 10 μL of 7α,25-dihydroxycholesterol (1 μM based on the curve) was added to each well of the microplate, in quadruplicate, at a concentration that elicited 80%-100% of the characteristic DMR signal intensity, in quadruplicate. The microplates were then placed on an Epic instrument for real-time DMR signal monitoring for 60 min. The response signal value was calculated based on the characteristic DMR response value of cells treated with 7α,25-dihydroxycholesterol. Results are shown in [Figure missing]. Figure 3 The experimental results showed that neither pungentine nor chitidine had DMR activating signals in CHO-K1-GPR183 cells (7.3% and -7.7%, both less than 40%), and neither had a desensitizing effect on the DMR of the GPR183 receptor agonist 7α,25-dihydroxycholesterol (-11% and -18%, both less than 50%), indicating that neither pungentine nor chitidine has GPR183 receptor activating activity. N-demethylnuciferine, on the other hand, had DMR activating signals in CHO-K1-GPR183 cells (53%), and had a desensitizing effect on the DMR of the GPR183 receptor agonist 7α,25-dihydroxycholesterol (57%), indicating that N-demethylnuciferine has GPR183 receptor activating activity.

[0049] This invention establishes a label-free screening model for GPR183 based on label-free cell integrative pharmacology. This model has the advantages of not requiring fluorescent labeling and no additional indicators during the detection process, enabling efficient and reliable screening of commercially available synthetic and natural small molecule libraries or natural product extracts / mixtures to obtain highly active ligands for the GPR183 receptor, providing lead compounds for targeted therapy of GPR183-related diseases. Using this model, the GPR183 receptor agonist N-desmethylnuciferine was screened, providing lead compounds for drug development in GPR183-related diseases such as multiple sclerosis, colitis, type 1 diabetes, and neuroinflammatory / autoimmune diseases, cancer, and metabolic diseases.

[0050] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A label-free cell screening model for the GPR183 receptor, characterized in that: The label-free cell selection model was the stable cell line CHO-K1-GPR183, which highly expresses the GPR183 receptor.

2. The label-free cell screening model for the GPR183 receptor according to claim 1, characterized in that: The stable cell line CHO-K1-GPR183 was obtained by transfecting CHO-K1 cells with a plasmid containing GPR183.

3. The application of the label-free cell screening model for the GPR183 receptor as described in claim 1, characterized in that: Application of the label-free cell screening model in screening drugs for diseases related to the GPR183 receptor.

4. The application of the label-free cell screening model for the GPR183 receptor according to claim 3, characterized in that: The label-free cell screening model is used in screening drugs for the prevention and / or treatment of diseases by activating or antagonizing the GPR183 receptor.

5. The application of the label-free cell screening model for the GPR183 receptor according to claim 4, characterized in that: The disease is one or more of the following: multiple sclerosis, colitis, type 1 diabetes, neuroinflammatory / autoimmune diseases, cancer, and metabolic diseases.

6. A method for screening drugs for diseases closely related to the GPR183 receptor, characterized in that: Based on label-free cell integrative pharmacology, using the CHO-K1-GPR183 cell line that stably expresses GPR183 as described in claim 1, the agonist or antagonist activity of the test sample is determined by the similarity and specificity of the DMR signal spectrum of the test sample with the DMR characteristic signal spectrum of known standard agonists and / or antagonists, and drugs closely related to diseases of GPR183 receptor are screened.

7. The method for screening drugs for diseases closely related to the GPR183 receptor according to claim 6, characterized in that: The label-free cell integrated pharmacology technology utilizes a resonant waveguide grating (RWG) biosensor to convert the dynamic redistribution of intracellular components caused by drugs into a holistic, dynamic wavelength shift response signal. This signal is the response value (pm) of wavelength change, which is realized through an Epic optical biosensor 384 microplate.

8. The method for screening drugs for diseases closely related to the GPR183 receptor according to claim 6, characterized in that: The known standard agonist is 7α,25-dihydroxycholesterol; the known standard antagonist is NIBR189.

9. The method for screening drugs for diseases closely related to the GPR183 receptor according to any one of claims 6-8, characterized in that: The screened GPR183 receptor agonists were one or more of the compounds N-demethylnuciferine or pharmaceutically acceptable salts thereof, and the compounds had the following structural formulas:

10. The use of a compound in the preparation of a medicament for the prevention and / or treatment of diseases by activating the GPR183 receptor, characterized in that: The use of the following structural compounds in the preparation of medicaments for the prevention and / or treatment of diseases by activating GPR183 receptors;

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