Application of arachidonic acid as interleukin-5 receptor antagonist

Abstract

The invention discloses an application of arachidonic acid as an interleukin 5 receptor antagonist, and belongs to the technical field of biological medicines.The application comprises the steps that an interleukin 5 receptor serves as a target, and drug molecules capable of being combined with the interleukin 5 receptor in an FDA drug library are screened in a high-throughput mode; according to the invention, arachidonic acid is used as an antagonist of interleukin-5 receptors, the antagonism efficiency of the arachidonic acid is verified at an in-vitro molecular level, a molecule capable of antagonizing interleukin-5 receptors is screened out, the concentration of the screened arachidonic acid reaching the same antagonism effect as benralizumab is 86.61 pg / ml, and the application prospect of the arachidonic acid as the interleukin-5 receptor antagonist is proved.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically the application of arachidonic acid as an interleukin-5 receptor antagonist. Background Technology

[0002] Interleukin-5 (IL-5) is a multifunctional cytokine produced by immune system cells, primarily activated T cells, mast cells, and Th2 cells. When stimulated, these cells release IL-5 into the bloodstream, where it exerts its biological effects. IL-5 plays a crucial role in immune regulation and inflammatory responses: ① It promotes the growth, differentiation, recruitment, activation, and survival of eosinophils and basophils, thereby enhancing the body's immune response to parasitic infections and allergic diseases; ② It stimulates B cell proliferation and increases their antibody secretion capacity, thereby promoting antibody production and enhancing humoral immune responses; ③ It regulates hematopoietic function by promoting the proliferation and differentiation of hematopoietic stem cells in the bone marrow, maintaining the normal production and renewal of blood cells; ④ It promotes inflammatory responses, as IL-5 can stimulate inflammatory cells (such as eosinophils) to aggregate at the site of inflammation and release inflammatory mediators (such as histamine and leukotrienes), thus exacerbating the inflammatory response.

[0003] IL-5 exerts its biological effects by binding to the IL-5 receptor (IL-5R) on the surface of target cells. IL-5R consists of an α-chain and a β-chain. The α-chain triggers downstream signaling pathways upon binding to IL-5, while the β-chain participates in signal transduction processes, including Jak-Stat and MAPK, and can regulate biological processes such as cell proliferation, differentiation, and apoptosis. Due to its important role in immune responses and inflammatory reactions, IL-5 has broad application prospects in the medical field. For example, in the treatment of allergic diseases, drugs targeting IL-5 or IL-5R can inhibit eosinophil activity, thereby reducing allergic reactions and inflammation.

[0004] Given the crucial role of IL-5R in disease, biologics primarily composed of IL-5R inhibitors are an important component of IL-5R antagonists. Antibody drugs alleviate disease symptoms by blocking the interaction between IL-5 and IL-5R, inhibiting the accumulation of immune cells and inflammatory factors. Antibody therapy can replace hormone therapy; when combined with glucocorticoids, it can reduce the dosage of hormones and decrease the incidence of adverse drug reactions. Inflammatory cytokines, cell surface molecules, and their mediated signaling pathways participate in the pathological process of autoimmune diseases. Targeted biologics targeting cytokines, receptors, and signaling molecules have seen rapid development in recent years.

[0005] However, biologic antibody drugs carry certain side effects or treatment risks. Acute reactions following monoclonal antibody injection can be caused by various mechanisms, including acute anaphylaxis (IgE-mediated) and anaphylactic-like reactions, serum sickness, tumor lysis syndrome, and cytokine release syndrome. Clinical manifestations include local skin reactions at the injection site, fever, and flu-like syndrome. Humanization of antibody drugs does not completely eliminate immunogenicity; even the CDR region of humanized antibodies retains strong immunogenicity, generating anti-idiotype antibodies and leading to drug inactivation. Monoclonal antibodies and other biomolecules possess unique three-dimensional structures. The complex balance of interactions between amino acid functional groups and the external environment, both intramolecularly and intermolecularly, determines the folded structure. Because the folded structure is in a dynamic equilibrium, any factor altering this balance can cause structural changes, leading to instability in the macromolecule. Since the tertiary structure of biologics is susceptible to environmental physical stress, structural changes in monoclonal antibodies can occur at any stage of the production process, from initial protein expression to processing and storage. Most monoclonal antibodies are large and have limited ability to penetrate and accumulate tissues, likely confined to the interstitial space after injection. Biotherapies can reach the bloodstream via two pathways: through capillaries or lymphatic vessels. However, capillary absorption has been reported to rely on passive transport and is limited to compounds with molecular weights below 16 kDa. Therefore, most biotherapeutic drugs cannot be transported via the capillary route and instead rely on the lymphatic system. Furthermore, enzymatic degradation mechanisms within the body further reduce the pharmacokinetic properties of most types of protein drugs; proteins and monoclonal antibodies are prone to enzymatic degradation. Summary of the Invention

[0006] To overcome the shortcomings of the existing technology, this invention uses IL-5R as a target to conduct high-throughput screening of drug molecules in the FDA drug library that can bind to IL-5R, and verifies their antagonistic efficiency at the in vitro molecular level, thus screening out a molecule that can antagonize IL-5R, providing a new approach for studying the function of IL-5R.

[0007] The purpose of this invention is to protect the application of arachidonic acid as an interleukin-5 receptor antagonist.

[0008] Arachidonic acid, also known as cis-5,8,11,14-eicosatetraenoic acid, is an ω-6 polyunsaturated fatty acid. At room temperature, it is a colorless to pale yellow oily liquid, mainly found in animal organs, red meat, eggs, and some seafood.

[0009] Arachidonic acid (ARA) is an important structural ester and metabolic substrate in the human body. It also acts as a second messenger within cells, directly participating in intracellular signal transduction and influencing other signal transduction pathways to regulate cellular biological activities. Furthermore, ARA can lower cholesterol, relax vascular smooth muscle and coronary arteries, increase sinoatrial node frequency and myocardial contractility, playing a vital role in the cardiovascular system. Studies have found that ARA can enhance vaccine immunity, accumulating in lymph nodes and metabolizing into the active substance prostaglandin I2. Prostaglandin I2 activates the cAMP-PKA signaling pathway within B cells, enhancing the interaction between B cells and T cells, promoting germinal center formation, activating key enzymes, driving antibody class switching and affinity maturation, and ultimately producing more potent neutralizing antibodies.

[0010] In one specific embodiment, the interleukin-5 receptor antagonist was screened using the following method: (a) Establish an FDA-approved drug library: Customize a library containing at least 3,067 compound drugs, each dissolved in DMSO at a concentration of 1 mg / ml and stored. (b) Preparation of interleukin-5 receptor target: Custom-designed interleukin-5 receptor protein target with a purity of at least 95% and serially diluted to at least five concentration levels, including but not limited to 500 nM, 250 nM, 125 nM, 62.5 nM and 31.25 nM; (c) Molecular screening: The FDA drug library was assembled into a microfluidic chip and installed in a high-throughput screening analyzer. After verifying the chip quality using positive control samples, interleukin-5 receptor targets were injected sequentially according to the concentration gradient. The interaction with interleukin-5 receptor targets was detected, and the collected signal data was analyzed using data analysis software. The results were sorted according to the molecular affinity. (d) Candidate drug screening: In the results obtained in step (c), compound drugs with significant affinity for the interleukin-5 receptor target were identified, among which arachidonic acid was screened as a candidate drug with potential antagonistic effects. (e) Drug molecular level validation: Using an interleukin-5 receptor assay kit, with known interleukin-5 receptor monoclonal antibody drugs as positive controls, the antagonistic effect of arachidonic acid on interleukin-5 receptor was evaluated, and the antagonistic activity of arachidonic acid was verified by calculating the antagonism rate.

[0011] In one specific embodiment, the FDA-approved drug library may contain a wider variety of compound drugs to improve the diversity and accuracy of screening.

[0012] In one specific embodiment, the purity of the interleukin-5 receptor target was determined by methods such as SDS-PAGE.

[0013] In one specific embodiment, the high-throughput screening analyzer is a PlexArrayHTSPRi microarray analyzer or other instruments suitable for molecular screening.

[0014] In one specific embodiment, the antagonism rate in the drug molecule-level validation step is calculated as follows: Antagonism rate = 100% - (detected concentration / initial concentration) × 100%.

[0015] In one specific embodiment, the interleukin-5 receptor is shown in SEQ ID NO: 1, and its specific amino acid sequence is as follows: DLLPDEKISLLPPVNFTIKVTGLAQVLLQWKPNPDQEQRNVNLEYQVKINAPKEDDYETTRITESKCVTILHKGFSASVRTILQNDHSLLASSWASAELHAPPGSPGTSIVNLTCTTNTTEDNYSRLRSYQVSLHCTWLVGTDAPEDTQYFLYYRYGSW TEECQEYSKDTLGRNIACWFPRTFILSKGRDWLAVLVNGSSKHSAIRPFDQLFALHAIDQINPPLNVTAEIEGTRLSIQWEKPVSAFPIHCFDYEVKIHNTRNGYLQIEKLMTNAFISIIDDLSKYDVQVRAAVSSMCREAGLWSEWSQPIYVGNDE.

[0016] Another object of the present invention is to protect a pharmaceutical composition of an interleukin-5 receptor antagonist comprising arachidonic acid and one or more pharmaceutically acceptable carriers or excipients.

[0017] In one specific embodiment, the carrier comprises liposomes or cyclodextrin complexes, and the excipients comprise antioxidants and metal ion chelates.

[0018] In one specific embodiment, the antioxidant is one or more of resveratrol, α-tocopherol, pterostilbene, and probucol, and the metal ion chelate is selected from one or more of disodium ethylenediaminetetraacetate or citric acid.

[0019] In one specific embodiment, the antioxidant is resveratrol and α-tocopherol.

[0020] In one specific embodiment, the carrier is a liposome-cyclodextrin complex.

[0021] Another object of the present invention is to protect a drug-loaded liposome of an interleukin-5 receptor antagonist, wherein the drug-loaded liposome, by molar ratio, comprises 10-15 parts arachidonic acid, 0.1-0.5 parts resveratrol, 0.01-0.05 parts α-tocopherol, 300-500 parts liposome source, and 80-100 parts anionic cyclodextrin. The liposome source comprises 100-200 parts hydrogenated soybean phosphatidylcholine, 50-70 parts (2,3-dioleopropyl)trimethylammonium chloride, 80-120 parts cholesterol, 15-30 parts hydrogenated sphingomyelin, 10-15 parts distearate phosphatidylethanolamine methoxy polyethylene glycol, and 5-10 parts dipalmitoylphosphatidylglycerol. The anionic cyclodextrin comprises sodium sulfobutylbetacyclodextrin.

[0022] The drug-loaded liposomes are prepared by using a thin-film-hydration extrusion-cyclodextrin coating method.

[0023] The film-hydration extrusion-cyclodextrin coating method involves mixing a liposome source, a drug active component, and a first organic solvent to obtain a first mixed solution. The first organic solvent in the first mixed solution is then evaporated to remove it, forming a film. The film is then mixed with an hydration solution to obtain a second mixed solution. After hydration, the solution is extruded and purified. Finally, the drug-loaded liposomes are coated with anionic cyclodextrin.

[0024] The first organic solvent can be a solvent capable of dissolving various raw materials, preferably at least one of anhydrous ethanol, chloroform and methanol, and most preferably anhydrous ethanol.

[0025] Arachidonic acid has certain application prospects as an interleukin-5 receptor antagonist. However, the molecular structure of arachidonic acid contains four cis double bonds, which are easily oxidized by oxygen or metal ions at room temperature, leading to a decrease in efficacy. Furthermore, arachidonic acid has extremely poor water solubility, cannot be directly dissolved by intravenous injection, has low oral bioavailability, and also has the problem of a short half-life. Therefore, this invention makes it into liposomes, giving it the advantages of oxygen barrier and water solubility, and can prolong its circulation time.

[0026] Beneficial effects

[0027] Arachidonic acid is an important ω-6 polyunsaturated fatty acid that can be used as an antagonist of IL-5R, potentially replacing antibody drugs in antagonizing IL-5R. It boasts advantages such as low toxicity, minimal side effects, stable drug molecular structure, and good pharmacokinetic properties. Forming it into liposomes not only endows it with oxygen-barrier and water-soluble advantages but also prolongs its circulation time. Furthermore, this invention provides a drug-loaded liposome. By mixing the active drug molecule with a liposome source to form a film, followed by hydration and extrusion, a positively charged liposome primary product is obtained. Then, anionic cyclodextrin is used to coat its surface through charge neutralization, making the liposome shell more stable. This solves the problems of poor stability of conventional liposomes and low encapsulation efficiency of simple cyclodextrin, resulting in a drug-loaded liposome with oxygen barrier properties, good water solubility, and long-lasting stability. Attached Figure Description

[0028] Figure 1 Arachidonic acid signal curves were collected when IL-5R protein concentrations were 500.0 nM, 250.0 nM, 125.0 nM, 62.5 nM and 31.2 nM.

[0029] Figure 2 This is the molecular structure diagram of arachidonic acid.

[0030] Figure 3 The graph shows the efficacy analysis of the positive control benazepril at concentrations of 0, 3, 6, 12, 24, 48, and 96 μg / ml.

[0031] Figure 4 The chromatograms show the efficacy of arachidonic acid at concentrations of 0, 1.19, 4.74, 18.97, 75.9, 303.6, and 1214.39 μg / ml. Detailed Implementation

[0032] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0033] Unless otherwise specified, the experimental methods used in the embodiments are conventional methods, and the materials and reagents used are commercially available unless otherwise specified.

[0034] Unless otherwise specified, all components and raw materials used in the embodiments and comparative examples of this invention are commercially available, and the same type of components and raw materials are used in each parallel experiment.

[0035] Example 1

[0036] This embodiment focuses on high-throughput screening of FDA drug library chips targeting IL-5R. The experimental steps are as follows: The equipment and reagents used in the experiment were as follows: FDA drug library chip (purchased from Selleck Chemicals), interleukin-5 receptor protein (purchased from Bispecies Biosciences), PBST buffer (purchased from Thermo Fisher Scientific), and Gly-HCl buffer (purchased from Thermo Fisher Scientific). In one specific embodiment, the experimental steps for high-throughput screening of FDA drug library chips using IL-5R as the target are as follows: (1) Take out the prepared FDA drug library chip, place it in 1% BSA solution for 2 hours, wash the chip with pure water and dry it before assembling it into a microfluidic chip.

[0037] (2) 1 mg / ml of interleukin-5 receptor (IL-5R) protein was serially diluted with PBST buffer.

[0038] (3) The microfluidic chip was installed in the PlexArrayHTSPRi microarray analyzer. First, a positive sample of FKBP12 with a concentration of 100 nM was injected at a flow rate of 1 μl / sec for 300 sec. The positive response signal of rapamycin was detected to verify the chip quality.

[0039] (4) Wash with PBST buffer at a flow rate of 1 μl / s for 300 sec. Inject the protein of the gradient concentration sequentially at a flow rate of 1 μl / s for 300 sec. Dissociate with PBST buffer at a flow rate of 1 μl / s for 300 sec. Repeat the above steps until all protein concentrations have been injected. Collect signal data.

[0040] (5) The chip was regenerated and stored after binding for 120s and dissociating for 180s with Gly-HCl buffer (pH=2.0) at a flow rate of 1μl / s.

[0041] (6) The collected signal data were analyzed using the data analysis software DataProcessorStandAlone, and the results were sorted by molecular affinity.

[0042] The experimental results are as follows Figure 1 As shown in the results, the detected binding signal becomes stronger with increasing protein concentration. Arachidonic acid, a small molecule compound numbered S6185, was selected as the candidate molecule due to its high affinity among all small molecules. Further research will be conducted using this compound as a candidate. The molecular structure of arachidonic acid is shown below. Figure 2 As shown.

[0043] Example 2

[0044] This embodiment analyzes the efficacy of arachidonic acid in antagonizing IL-5R and compares it with the efficacy of the positive control benazepril antagonizing IL-5R.

[0045] The efficacy comparison of benazepril antagonism against IL-5R is as follows: The equipment and reagents used in the experiment are as follows: human interleukin-5 receptor (IL-5R), ELISA kit (purchased from Shenzhen Zike Biotechnology Co., Ltd.), and benalizumab (purchased from MedChemexpress Biotechnology Co., Ltd., USA).

[0046] The experimental procedure for the positive control benaribizumab antagonizing IL-5R is as follows: (1) Add 50 μl of 100 pg / ml IL-5R protein to an EP tube, add different concentrations of benazepril 1, and incubate at 37°C for 1 h.

[0047] (2) Add 100 μl of protein small molecule mixture or standard and 100 μl of HRP-labeled IL-5R antibody to the pre-coated microwells, incubate at 37°C for 1 h, discard the solution, wash 5 times, and develop TMB color.

[0048] (3) Measure the absorbance at 450 nm using an enzyme-linked immunosorbent assay (ELISA) reader, establish a standard curve, and then calculate the detection concentration.

[0049] The antagonism rate is calculated based on the detected concentration: Antagonism rate = 100% - (detected concentration / initial concentration).

[0050] The results are shown in Table 1. A linear fit was performed on the logarithm of benalizumab drug concentration and the antagonism rate, and the results are as follows: Figure 3 As shown, for reference Figure 3 The efficacy analysis of 30 mg benalizumab administered subcutaneously showed that the antagonistic effect reached 16.93% when the concentration of the monoclonal antibody was 6.00 μg / ml (the concentration used).

[0051] Table 1. Results of the dose-effect analysis of benazepril.

[0052] The efficacy of arachidonic acid in antagonizing IL-5R was analyzed: The equipment and reagents used in the experiment were as follows: human interleukin-5 receptor (IL-5R), ELISA kit (purchased from Shenzhen Zike Biotechnology Co., Ltd.), benazepril zumab (purchased from MedChemexpress Biotechnology Co., Ltd., USA), and arachidonic acid (purchased from MedChemexpress Biotechnology Co., Ltd., USA).

[0053] The experimental procedures for analyzing the efficacy of arachidonic acid in antagonizing IL-5R are as follows: (1) Add 50 μl of 120 pg / ml IL-5R protein to an EP tube, add different concentrations of arachidonic acid, and incubate at 37°C for 1 h.

[0054] (2) Add 100 μl of protein small molecule mixture or standard and 100 μl of HRP-labeled IL-5R antibody to the pre-coated microwells, incubate at 37°C for 1 h, discard the solution, wash 5 times, and develop TMB color.

[0055] (3) The absorbance at 450 nm was measured using an ELISA reader. After establishing a standard curve, the detection concentration was calculated. The antagonism rate was calculated based on the detection concentration: antagonism rate = 100% - (detection concentration / initial concentration). The results are shown in Table 2. A linear fit was performed on the logarithm of the drug concentration and the antagonism rate of S6185 arachidonic acid, and the results are as follows: Figure 4 As shown, the dose-effect analysis results revealed that arachidonic acid achieved an antagonistic effect equivalent to benazepril at a concentration of 86.61 pg / ml, demonstrating the promising application of arachidonic acid as an interleukin-5 receptor antagonist.

[0056] Table 2. Effect analysis results of arachidonic acid

[0057] Example 3

[0058] A drug-loaded liposome, by molar amounts, comprises 10-15 parts arachidonic acid, 0.1-0.5 parts resveratrol, 0.01-0.05 parts α-tocopherol, 300-500 parts liposome source, and 60-100 parts anionic cyclodextrin. The liposome source comprises 100-200 parts hydrogenated soybean phosphatidylcholine, 50-70 parts (2,3-dioleopropyl)trimethylammonium chloride, 80-120 parts cholesterol, 15-30 parts hydrogenated sphingomyelin, 10-15 parts distearate phosphatidylethanolamine methoxy polyethylene glycol, and 5-10 parts dipalmitoylphosphatidylglycerol. The anionic cyclodextrin is sodium sulfobutylbetacyclodextrin. The preparation method of the drug-loaded liposome is as follows: (1) 0.2 mmol of hydrogenated soybean phosphatidylcholine, 0.06 mmol of (2,3-dioleoxypropyl)trimethylammonium chloride, 0.1 mmol of cholesterol, 0.02 mmol of hydrogenated sphingomyelin, 0.012 mmol of distearyl phosphatidylethanolamine methoxy polyethylene glycol, and 0.008 mmol of dipalmitoyl phosphatidylglycerol were mixed with 15 ml of chloroform / methanol (9:1 v / v). After dissolution, 0.012 mmol of arachidonic acid, 0.0003 mmol of resveratrol, and 0.00005 mmol of α-tocopherol were added to obtain the first mixed solution. The first organic solvent in the first mixed solution was removed by rotary evaporation at 55 °C and vacuum for 30 min to form a thin film.

[0059] (2) Prepare an hydration solution containing 10 mmol of 4-hydroxyethylpiperazine ethanesulfonic acid, 150 mmol of NaCl, pH 7.4, 0.005 mmol of EDTA-Na2 and 1 wt% of sucrose, and preheat it to 55°C. Pour the solution into a round-bottom bottle, mix the membrane with 10 ml of the hydration solution, and hydrate it at 55°C and 50 rpm for 15 min to obtain a milky white suspension. Preheat the extruder to 55°C and pass the suspension through polycarbonate membranes with pore sizes of 0.4 µm, 0.2 µm and 0.1 µm three times each, at a pressure of 200–400 psi, to obtain crude liposomes.

[0060] (3) The crude liposome product was purified by using a 100kDa ultrafiltration centrifuge tube. Each time, 10 mL of fresh 4-hydroxyethylpiperazine ethanesulfonic acid-sodium chloride buffer was added to wash and filter three times to obtain liposomes with positive surface charge.

[0061] (4) Dissolve 0.08 mmol of sodium sulfobutyl betacyclodextrin in 0.5 ml of water and slowly drip it into liposomes preheated at 55 °C with a positively charged surface. While adding the liposomes, gently shake them and incubate for 30 min. Remove free cyclodextrin by ultrafiltration three times to obtain drug-loaded liposomes. The particle size of the drug-loaded liposomes was 127 nm, with an encapsulation efficiency of 75%. After storage at 4 °C for 16 days, the particle size was retested and remained stable at 127 nm, indicating good stability.

[0062] Example 4

[0063] Compared with Example 3, the difference is that the preparation method of drug-loaded liposomes does not include step (4), that is, it is only liposomes. The particle size of the drug-loaded liposomes obtained is 115nm and the encapsulation efficiency is 62.3%. However, after 16 days, the encapsulation efficiency dropped to 55% and the stability was slightly lower.

[0064] The preferred embodiments of the present invention disclosed above are merely illustrative of the invention. These preferred embodiments do not exhaustively describe all details, nor do they limit the invention to the specific implementations described. Clearly, many modifications and variations can be made based on the content of this specification. This specification selects and specifically describes these embodiments to better explain the principles and practical applications of the invention, thereby enabling those skilled in the art to better understand and utilize the invention. The invention is limited only by the claims and their full scope and equivalents.

Claims

1. Application of arachidonic acid as an interleukin-5 receptor antagonist.

2. The application of arachidonic acid as an interleukin-5 receptor antagonist according to claim 1, characterized in that, The arachidonic acid was obtained through the following screening method: (a) Establish an FDA-approved drug library: Customize a library containing at least 3,067 compound drugs, each dissolved in DMSO at a concentration of 1 mg / ml and stored. (b) Preparation of interleukin-5 receptor target: Custom-designed interleukin-5 receptor protein target with a purity of at least 95% and serially diluted to at least five concentration levels, including but not limited to 500 nM, 250 nM, 125 nM, 62.5 nM and 31.25 nM; (c) Molecular screening: The FDA drug library was assembled into a microfluidic chip and installed in a high-throughput screening analyzer. After verifying the chip quality using positive control samples, interleukin-5 receptor targets were injected sequentially according to the concentration gradient. The interaction with interleukin-5 receptor targets was detected, and the collected signal data was analyzed using data analysis software. The results were sorted according to the molecular affinity. (d) Candidate drug screening: In the results obtained in step (c), compound drugs with significant affinity for the interleukin-5 receptor target were identified, among which arachidonic acid was screened as a candidate drug with potential antagonistic effects. (e) Drug molecular level validation: Using an interleukin-5 receptor assay kit, with known interleukin-5 receptor monoclonal antibody drugs as positive controls, the antagonistic effect of arachidonic acid on interleukin-5 receptor was evaluated, and the antagonistic activity of arachidonic acid was verified by calculating the antagonism rate.

3. The application of arachidonic acid as an interleukin-5 receptor antagonist according to claim 2, characterized in that: The FDA-approved drug library can contain a wider variety of compound drugs to improve the diversity and accuracy of screening; the purity of the interleukin-5 receptor target is determined by SDS-PAGE; the high-throughput screening analyzer is a PlexArrayHTSPRi microarray analyzer; in the drug molecule-level validation step, the antagonism rate is calculated as follows: antagonism rate = 100% - (detection concentration / initial concentration) × 100%.

4. The application of arachidonic acid as an interleukin-5 receptor antagonist according to claim 2, characterized in that: The interleukin-5 receptor is shown in SEQ ID NO:

1.

5. A pharmaceutical composition of an interleukin-5 receptor antagonist, characterized in that, It contains arachidonic acid and one or more pharmaceutically acceptable carriers or excipients; the carriers include liposomes, cyclodextrin complexes, or liposome-cyclodextrin complexes, and the excipients include antioxidants and metal ion chelates.

6. The pharmaceutical composition according to claim 5, characterized in that, The antioxidant is one or more of resveratrol, α-tocopherol, pterostilbene, and probucol, and the metal ion chelate is selected from one or more of disodium ethylenediaminetetraacetate or citric acid.

7. The pharmaceutical composition according to claim 5, characterized in that, The antioxidants are resveratrol and α-tocopherol, and the carrier is a liposome-cyclodextrin complex.

8. A drug-loaded liposome of an interleukin-5 receptor antagonist, characterized in that, The drug-loaded liposomes, by weight, comprise 10-15 parts arachidonic acid, 0.1-0.5 parts resveratrol, 0.01-0.05 parts α-tocopherol, 300-500 parts liposome source, and 80-100 parts anionic cyclodextrin. The liposome source comprises 100-200 parts hydrogenated soybean phosphatidylcholine, 50-70 parts (2,3-dioleoxypropyl)trimethylammonium chloride, 80-120 parts cholesterol, 15-30 parts hydrogenated sphingomyelin, 10-15 parts distearyl phosphatidylethanolamine methoxy polyethylene glycol, and 5-10 parts dipalmitoyl phosphatidylglycerol. The anionic cyclodextrin comprises sodium sulfobutyl betacyclodextrin.

9. The method for preparing drug-loaded liposomes according to claim 8, characterized in that, Using a film-hydration extrusion-cyclodextrin coating method, liposome source, active pharmaceutical ingredient and first organic solvent are mixed to obtain a first mixed solution. The first organic solvent in the first mixed solution is removed by evaporation to form a film. The film is mixed with an aqueous solution to obtain a second mixed solution. After hydration, the solution is extruded and purified. Finally, the drug-loaded liposomes are coated with anionic cyclodextrin.

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