Human immunoglobulin E antagonist, screening method and application

Abstract

The present invention relates to the field of pharmaceutical chemistry technology, and in particular to a human immunoglobulin E antagonist, a screening method and an application. The screening method comprises the following steps: taking IgE as a target, high-throughput screening of drug molecules that can bind to IgE in the FDA drug library; in vivo testing of the antagonistic efficiency of the above-screened drug molecules, and screening out molecules that can antagonize IgE according to the expected antagonistic efficiency. The present invention uses IgE as a target, high-throughput screening of drug molecules that can bind to IgE in the FDA drug library, and verifies its antagonistic efficiency at the in vitro molecular level, thereby screening out a molecule that can antagonize IgE, and the antagonistic efficiency of andrographolide potassium succinate screened out has a significant antagonistic efficiency compared to ordinary antagonists.

Description

Technical Field

[0001] The present invention relates to the technical field of medicinal chemistry, and in particular to a human immunoglobulin E antagonist, a screening method and an application thereof. Background Art

[0002] Human immunoglobulin E (IgE) refers to the production of antibodies that can specifically bind to the corresponding antigens when B lymphocytes are transformed into plasma cells under antigen stimulation. It is an immunoglobulin that plays an important role in the human immune system. It has a high affinity for eosinophils and mast cells and is the main antibody that causes type I allergic reactions. In normal human blood, the content of IgE is extremely low, accounting for about 0.002% of the total immunoglobulin in serum. Due to the important role of IgE in immune response and inflammatory response, it has broad application prospects in the medical field.

[0003] Under normal physiological function, IgE is highly specific and mainly acts as a key molecule that mediates allergic reactions. It can recognize and bind to allergens (pollen, dust mites, animal dander, food, drugs, etc.) that the body comes into contact with. IgE will quickly recognize and bind to these allergens, thereby activating immune cells such as mast cells and eosinophils, triggering a series of immune responses, including inflammatory mediators such as histamine release, and ultimately leading to allergic reactions such as skin redness, sneezing, or asthma. Under normal conditions, the level of IgE remains within a relatively stable range to ensure that it responds to potentially harmful substances without triggering excessive or unnecessary immune responses. In allergic diseases such as allergic rhinitis and allergic asthma, IgE mainly binds tightly to the high-affinity receptor FcεR1 on the surface of mast cells and basophils. The bound IgE-FcεR1 complex causes changes in the receptor conformation, thereby activating protein kinases in the cell (such as Src family kinases and Syk kinases), further triggering changes in downstream signaling pathways, such as the MAPK signaling pathway and the NF-κB signaling pathway.

[0004] Given the key role of IgE in a variety of diseases, biological agents composed mainly of IgE targeted inhibitors are an important component of IgE antagonists. Antibody drugs block IgE-mediated allergic reactions by blocking the binding of IgE to high-affinity receptors (FcεRI) on the surface of effector cells (such as mast cells and eosinophils). This mechanism of action can effectively reduce the activation of mast cells and eosinophils, thereby reducing the release of inflammatory mediators, thereby alleviating allergic symptoms. Currently approved IgE inhibitors for clinical use include omalizumab, levofloxacin, etc.

[0005] Biologic antibody drugs have certain side effects or treatment risks. Acute reactions after injection of monoclonal antibodies can be caused by a variety of mechanisms, including acute allergic (IgE-mediated) and anaphylactic-like reactions to monoclonal antibodies, serum sickness, tumor lysis syndrome, and cytokine release syndrome. Clinical manifestations include local skin reactions at the injection site, fever, and flu-like syndrome. The humanization of antibody drugs does not completely eliminate immunogenicity. Even the CDR region of the humanized antibody still has strong immunogenicity, producing anti-idiotypic antibodies and causing drug failure.

[0006] Although existing IgE antibody drug therapies have achieved remarkable results in the treatment of IgE-mediated allergic diseases, they still have some obvious disadvantages. First, these antibody drugs may cause serious side effects, including but not limited to allergic reactions, immunosuppression, and serum sickness, posing a potential threat to the patient's health. Secondly, IgE antibody drugs have a high treatment risk, especially when used for a long time or in high doses, which may cause more complex immune problems and physical damage. In addition, these drugs also have deficiencies in stability and pharmacokinetic properties, which may lead to unstable efficacy and increase the difficulty and uncertainty of treatment. Therefore, it is particularly important to develop a new drug or antagonist that can improve the shortcomings of IgE antibody drug therapy. Summary of the invention

[0007] In view of the deficiencies in the prior art, the present invention provides a human immunoglobulin E antagonist, a screening method and an application.

[0008] In one aspect, the present invention provides a human immunoglobulin E antagonist, which is andrographolide succinate potassium salt.

[0009] In a further preferred embodiment, the andrographolide potassium succinate is obtained by the following screening method:

[0010] (a) Establishment of an FDA-approved drug library: A library containing at least 3067 compound drugs was customized, each drug was dissolved in DMSO at a concentration of 1 mg / ml and stored;

[0011] (b) preparing human immunoglobulin E target: customizing a human immunoglobulin E protein target with a purity of at least 95%, and gradiently diluting it into at least five concentration levels, including but not limited to 500 nM, 250 nM, 125 nM, 62.5 nM and 31.25 nM;

[0012] (c) Molecular screening: The FDA drug library is assembled into a microfluidic chip and installed in an appropriate high-throughput screening analyzer. After the chip quality is verified using a positive control sample, the human immunoglobulin E target is injected in sequence according to a concentration gradient. The interaction with the human immunoglobulin E target is detected, and the collected signal data is analyzed using data analysis software, and the results are ranked according to the molecular affinity;

[0013] (d) Candidate drug screening: From the results obtained in step (c), a compound drug having a significant affinity reduction effect on the human immunoglobulin E target is identified, among which andrographolide potassium succinate is screened as a candidate drug with potential antagonistic effect;

[0014] (e) Drug molecular level verification: A human immunoglobulin E detection kit was used, with known human immunoglobulin E monoclonal antibody drugs as positive controls, to evaluate the antagonistic effect of andrographolide potassium succinate on human immunoglobulin E. The antagonistic activity of andrographolide potassium succinate was verified by calculating the antagonism rate.

[0015] In a further preferred embodiment, the andrographolide potassium succinate is confirmed to have a significant affinity-reducing effect in step (d), and shows an antagonism rate at least equivalent to that of the positive control drug through ELISA detection in step (e).

[0016] In a further preferred embodiment, the FDA-approved drug library may contain more types of compound drugs to improve the diversity and accuracy of screening.

[0017] The purity of the human immunoglobulin E target can be determined by SDS-PAGE or other methods to ensure that it meets the screening requirements.

[0018] In a further preferred embodiment, the high-throughput screening analyzer may be a PlexArray HT SPRi microarray analyzer or other instruments suitable for molecular screening;

[0019] In the drug molecule level verification step, the antagonism rate is calculated as follows: antagonism rate = 100% - (detection concentration / initial concentration) x 100%.

[0020] In a further preferred embodiment, in step (e), the IL-5R monoclonal antibody drug omalizumab is used as a positive control, and the IL-5R amino acid sequence is shown in SEQ ID NO: 1.

[0021] Another aspect of the present invention provides an antagonist for improving the side effects of IgE antibody drug therapy, wherein the main component of the antagonist is andrographolide potassium succinate.

[0022] Another aspect of the present invention provides a use of andrographolide potassium succinate as a human immunoglobulin E antagonist.

[0023] Another aspect of the present invention provides an antiallergic pharmaceutical composition containing andrographolide potassium succinate.

[0024] Another aspect of the present invention provides a method for treating or preventing allergic diseases using andrographolide potassium succinate. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The above and other purposes, features and advantages of the present invention will become more apparent by more specific description of the preferred embodiments of the present invention shown in the accompanying drawings. The same reference numerals indicate the same parts in all the accompanying drawings, and the accompanying drawings are not deliberately scaled to the actual size, but the focus is on illustrating the main purpose of the present invention.

[0026] Figure 1 The signal curves of andrographolide potassium succinate collected when the IgE protein concentrations were 500.0nM, 250.0nM, 125.0nM, 62.5nM and 31.2nM, respectively.

[0027] Figure 2 This is the molecular structure diagram of andrographolide potassium succinate.

[0028] Figure 3 This is a dose-effect analysis diagram of the positive control omalizumab at concentrations of 0μg / ml, 18.75μg / ml, 37.5μg / ml, 75μg / ml, 150μg / ml, 300μg / ml, and 600μg / ml.

[0029] Figure 4 This is a dose-effect analysis diagram of andrographolide potassium succinate at 0μg / L, 151.56μg / L, 303.13μg / L, 606.25μg / L, 1212.5μg / L, 2425μg / L and 4850μg / L. DETAILED DESCRIPTION

[0030] To facilitate understanding of the present invention, a more comprehensive description of the present invention will be given below with reference to the relevant drawings.

[0031] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art. The terms used in the specification herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0032] The present invention provides a method for screening an immunoglobulin E antagonist, comprising the following steps:

[0033] First, with IgE as the target, drug molecules that can bind to IgE are searched in the high-throughput screening FDA drug library; then, these drug molecules are tested for antagonistic efficiency in vivo to screen out molecules that can effectively antagonize IgE. In the specific process, 3067 customized FDA-approved compound drugs and IgE protein targets with measured purity are used for screening through microfluidic chips and PlexArray HT SPRi microarray analyzers, and finally, small molecule compounds that meet the requirements, namely andrographolide potassium succinate, are screened out through data analysis software. Then, the antagonistic effect of andrographolide potassium succinate is detected and evaluated using the immunoglobulin E ELISA detection kit. The IL-5R monoclonal antibody drug omalizumab is also used as a positive control during the evaluation process. The entire screening process includes steps such as incubation of IgE and andrographolide potassium succinate, immune response and color development, absorbance determination and concentration calculation.

[0034] More specifically, the steps include:

[0035] (a) FDA drug library establishment: 3067 FDA-approved compound drugs were customized and stored in 1 mg / ml DMSO.

[0036] (b) IgE target preparation. IgE protein target was customized, and its purity was determined to be 95% by SDS-PAGE. IgE was gradient diluted with deionized water to 500 nM, 250 nM, 125 nM, 62.5 nM and 31.25 nM.

[0037] (c) Molecular screening with IgE as the target. The FDA drug library chip was assembled into a microfluidic chip and installed in the PlexArray HT SPRi microarray analyzer. The positive sample rapamycin was tested to verify the chip quality. The proteins to be tested were injected in sequence according to the concentration gradient. Finally, the collected signal data was analyzed using the data analysis software Data Processor Stand Alone, and the results were ranked by molecular affinity.

[0038] (d) Drug molecular level verification. The antagonistic effect of andrographolide potassium succinate on IgE was evaluated using the human immunoglobulin E (IgE) ELISA test kit (Shenzhen Zike), with the IgE monoclonal antibody drug omalizumab as a positive control. The absorbance was detected using an enzyme marker, and the antagonism rate was calculated based on the detected concentration, antagonism rate = 100% - (detection concentration / initial concentration).

[0039] The amino acid sequence of IL-5R is shown in SEQ ID NO:1.

[0040] SEQ ID NO:1

[0041] 1:DLLPDEKISLLPPVNFTIKVTGLAQVLLQWKPNPDQEQRNVNLEYQVKINAPKEDDYETR

[0042] 61:ITESKCVTILHKGFSASVRTILQNDHSLLASSWASAELHAPPGSPGTSIVNLTCTTNTTE

[0043] 121:DNYSRLRSYQVSLHCTWLVGTDAPEDTQYFLYYRYGSWTEECQEYSKDTTLGRNIACWFPR

[0044] 181:TFILSKGRDWLAVLVNGSSKHSAIRPFDQLFALHAIDQINPPLNVTAEIEGTRLSIQWEK

[0045] 241:PVSAFPIHCFDYEVKIHNTRNGYLQIEKLMTNAFISIIDDLSKYDVQVRAAVSSMCREAG

[0046] 301:LWSEWSQPIYVGNDE

[0047] In a preferred embodiment, the steps of high-throughput screening of the FDA drug library chip using IgE as a target are as follows:

[0048] (1) The prepared FDA drug library chip was taken out and placed in a 1% BSA solution for blocking for 2 h. The chip was washed with pure water and dried before being assembled into a microfluidic chip.

[0049] (2) Dilute the 1 mg / ml human immunoglobulin E (IgE) protein in PBST buffer in multiple dilutions.

[0050] (3) The microfluidic chip was installed in the PlexArray HT SPRi microarray analyzer, and a 100 nM FKBP12 positive sample was first circulated at a flow rate of 1 μl / sec for 300 sec. The positive response signal of rapamycin was detected to verify the quality of the chip.

[0051] (4) The cells were washed with PBST buffer at a flow rate of 1 μl / s for 300 sec, and the proteins to be tested were injected in gradient concentrations at a flow rate of 1 μl / sec for 300 sec. The cells were dissociated with PBST buffer at a flow rate of 1 μl / s for 300 sec. The above steps were repeated until all concentrations of proteins were injected and the signal data were collected.

[0052] (5) The chip was regenerated with Gly-HCl buffer (pH=2.0) at a flow rate of 1 μl / s, with binding for 120 s and dissociation for 180 s, and then stored.

[0053] (6) Use the data analysis software Data Processor Stand Alone to analyze the collected signal data and rank the results according to the molecular affinity.

[0054] Results reference Figure 1 and Figure 2 , and screened out the small molecule compound Dehydroandrographolide Succinate Potasium Salt numbered S3774. The results show that as the protein concentration increases, the detected binding signal becomes stronger, and its affinity ranks first among all small molecules. It is used as a candidate molecule for subsequent research.

[0055] In a preferred embodiment, the steps for analyzing the IgE antagonistic efficacy of the positive control omalizumab are as follows: (1) Add 50 μl of 10 pg / ml IgE protein to an EP tube, add different concentrations of omalizumab respectively, and incubate at 37°C for 1 hour.

[0056] (2) Add 100 μl of protein small molecule mixture or standard and 100 μl of HRP-labeled IgE antibody to the coated microwells pre-coated with IgE antibody, in turn, incubate at 37°C for 1 h, discard the solution, wash 5 times, and develop color with TMB.

[0057] (3) Use an enzyme-labeled instrument to measure the absorbance at 450 nm and calculate the detection concentration after establishing a standard curve.

[0058] The antagonism rate was calculated according to the test concentration, antagonism rate = 100% - (test concentration / initial concentration).

[0059] 3 Experimental results

[0060] Please refer to Figure 3 As shown in Table 1, the results of the dose-effect analysis of omalizumab showed that the antagonistic effect reached 22.96% at a concentration of 120 μg / ml.

[0061] Table 1:

[0062]

[0063] In a preferred embodiment, the steps for analyzing the efficacy of andrographolide potassium succinate against IgE are as follows:

[0064] (1) Add 50 μl of 100 pg / ml IgE protein to an EP tube, add different concentrations of andrographolide potassium succinate, and incubate at 37°C for 1 h.

[0065] (2) Add 100 μl of protein small molecule mixture or standard and 100 μl of HRP-labeled IL-5R antibody to the coated microwells pre-coated with IL-5R antibody, in turn, incubate at 37°C for 1 h, discard the solution, wash 5 times, and develop color with TMB.

[0066] (3) Use an enzyme-labeled instrument to measure the absorbance at 450 nm and calculate the detection concentration after establishing a standard curve.

[0067] The antagonism rate was calculated according to the test concentration, antagonism rate = 100% - (test concentration / initial concentration).

[0068] Please refer to Figure 4 As shown in Table 2, the results of the dose-effect analysis of S3774 andrographolide potassium succinate showed that the concentration of the antagonist with the same antagonistic effect was 0.11 μg / ml.

[0069] Table 2:

[0070]

[0071] The above materials and reagents include:

[0072] FDA drug library chip was purchased from Selleck Chemicals, human immunoglobulin E was purchased from Biopsy Biotech, PBST buffer was purchased from Thermo Fisher Scientific, Gly-HCl buffer was purchased from Thermo Fisher Scientific, omalizumab was purchased from MedChemexpress Biotech, and andrographolide potassium succinate was purchased from MedChemexpress Biotech, USA.

[0073] In summary, the present invention uses IgE as a target, high-throughput screening of drug molecules that can bind to IgE in the FDA drug library, and verifies its antagonistic efficiency at the in vitro molecular level, screens out a molecule that can antagonize IgE, and the antagonistic efficiency of andrographolide potassium succinate screened out has a significant antagonistic efficiency compared to common antagonists. First, the drug andrographolide potassium succinate screened out by the present invention is an important natural active product, which effectively avoids side effects such as acute allergic reactions and ensures the safety of patients. Secondly, by optimizing the drug structure, this technology reduces the immunogenicity of the CDR region, significantly reduces the production of anti-idiotypic antibodies, thereby extending the shelf life of the drug. Furthermore, andrographolide potassium succinate has a stable structure and is not easily affected by environmental factors. Finally, andrographolide potassium succinate is excellent in pharmacokinetics, has a smaller volume and stronger tissue penetration ability, and can resist enzyme degradation in the organism, thereby improving the bioavailability and efficacy of the drug. These advantages make this technology have broad application prospects in the treatment of IgE-related diseases.

[0074] Andrographolide potassium succinate (LA), 5-[3-1,2-dithioheterocyclic)]pentanoic acid, was first isolated from bovine liver by Reed et al. in 1951. It is a trace element with multiple biological and pharmacological properties. It is a limiting essential nutrient required for cells to generate energy using energy substances such as sugars. It is the most powerful natural antioxidant in nature. Because andrographolide potassium succinate has an alkyl chain and a carboxyl end, it is both fat-soluble and water-soluble. It can pass through the blood-brain barrier and enter any tissue with almost no toxic side effects. In normal physiological functions, andrographolide potassium succinate participates in the tricarboxylic acid cycle as an important coenzyme, and also acts as an important repair factor in biological systems, removing oxygen free radicals and reactive oxygen species, and reducing the attack of free radicals on the body. Except for a small amount of andrographolide potassium succinate that can be synthesized through fatty acids and cysteine ​​in the body, the main andrographolide potassium succinate in the human body needs to be taken from food. Andrographolide potassium succinate that enters the body can be quickly absorbed into the blood and accumulated in various tissues in the body, such as the heart, liver, and skeletal muscle. Studies have found that andrographolide potassium succinate has been used to treat various chronic diseases, such as diabetes, liver fibrosis, and neurodegenerative diseases. In vitro supplementation of andrographolide potassium succinate can inhibit lipid oxidation in nerve tissue, prevent protein glycosylation, inhibit aldose reductase, and prevent glucose or galactose from being converted into sorbitol. It is often used clinically to treat and relieve peripheral neuropathy caused by late diabetes; it can also strengthen the function of liver activity, increase energy metabolism rate, and work with other antioxidants to reduce free radical damage to liver cells, reduce inflammatory response, maintain liver function, and have a clearing effect on fat accumulation caused by non-alcoholic fatty liver disease, reducing the burden on the liver. In addition, andrographolide potassium succinate is also a super antioxidant that can preserve and regenerate other antioxidants, balance blood sugar concentration, enhance the body's immune system, protect against damage by free radicals, participate in energy metabolism, increase the ability of other antioxidants to eliminate free radicals, and promote the recovery of insulin sensitivity.

[0075] In addition, andrographolide potassium succinate has a significant anti-inflammatory effect in inflammatory reactions, and its antioxidant capacity plays an important role in the anti-inflammatory process. Studies have shown that andrographolide potassium succinate can prevent the expression of ICAM-1 and the adhesion of endothelial human monocytes, inhibit the expression of NF-κB-dependent metalloproteinase-9, and reduce the expression of other inflammatory factors, such as IL-6 and TNF-α. In summary, andrographolide potassium succinate is a vitamin-like disulfide compound with rich biological activity and great application prospects.

[0076] On the other hand, the present invention relates to a new use of andrographolide potassium succinate, specifically using it as an antagonist of human immunoglobulin E (IgE). Andrographolide potassium succinate can effectively bind to IgE through its unique chemical structure and biological activity, thereby inhibiting IgE-mediated immune response and reducing the occurrence of allergic reactions.

[0077] On the other hand, the present invention provides an antiallergic pharmaceutical composition containing andrographolide potassium succinate, which further comprises a pharmaceutically acceptable carrier or excipient. The composition can effectively alleviate or prevent allergic reactions, including but not limited to hay fever, asthma and food allergies, through the antagonistic effect of andrographolide potassium succinate on IgE.

[0078] On the other hand, the present invention provides a method for preparing an antiallergic pharmaceutical composition containing andrographolide potassium succinate, which comprises mixing andrographolide potassium succinate with a pharmaceutically acceptable carrier or excipient.

[0079] On the other hand, the present invention provides a method for treating or preventing allergic diseases using andrographolide potassium succinate, the method comprising administering an effective amount of andrographolide potassium succinate or a pharmaceutical composition thereof to a patient. The method can effectively alleviate or prevent the symptoms of allergic diseases and improve the quality of life of patients by inhibiting IgE-mediated immune responses.

[0080] On the other hand, the present invention also relates to the use of andrographolide potassium succinate in the preparation of a drug for regulating the human immune system. Andrographolide potassium succinate can regulate the function of the immune system and reduce the production and activity of IgE through its antioxidant, anti-inflammatory and immunomodulatory properties, thereby helping to maintain the balance and stability of the immune system.

[0081] On the other hand, the present invention provides an immunomodulator containing andrographolide potassium succinate, which further comprises other immunomodulatory components and / or pharmaceutically acceptable carriers or excipients. The immunomodulator can effectively regulate the function of the immune system and reduce IgE-mediated immune responses through the synergistic effect of andrographolide potassium succinate and other components, thereby preventing or alleviating the occurrence of allergic diseases.

[0082] The present invention relates to an antagonist for improving the side effects of IgE antibody drug treatment. The main component of the antagonist is andrographolide potassium succinate, which is used to effectively reduce or eliminate the side effects caused by the IgE antibody drug when treating IgE-mediated allergic diseases, including but not limited to allergic reactions, immunosuppression, serum sickness, etc., thereby improving the safety of treatment and the tolerance of patients.

[0083] Furthermore, the antagonist also includes a pharmaceutically acceptable carrier, excipient or diluent to prepare a pharmaceutical preparation suitable for oral administration, injection or external use for the convenience of patients.

[0084] The present invention also relates to a pharmaceutical composition for treating IgE-mediated allergic diseases, which comprises any one of the above-mentioned antagonists and at least one IgE antibody drug, and is used for synergistically treating IgE-mediated allergic diseases, such as allergic rhinitis, allergic asthma, etc., while reducing the side effects of the IgE antibody drug.

[0085] The IgE antibody drugs include but are not limited to anti-IgE monoclonal antibodies, IgE receptor antagonists or IgE synthesis inhibitors, which act synergistically with andrographolide potassium succinate to improve the therapeutic effect and reduce the treatment risk.

[0086] The present invention also relates to a method for using the antagonist or pharmaceutical composition, which comprises administering the antagonist or pharmaceutical composition to a patient who needs to treat an IgE-mediated allergic disease under the guidance of a professional medical personnel, wherein the administration route includes but is not limited to oral administration, injection or external application, so as to alleviate allergic symptoms and improve the side effects of IgE antibody drug treatment.

[0087] Although the embodiments of the present application have been shown and described above, it can be understood that the above embodiments are exemplary and cannot be understood as limitations on the present application. Ordinary technicians in the field can change, modify, replace and modify the above embodiments within the scope of the present application.

Claims

1. A human immunoglobulin E antagonist, characterized in that The antagonist is andrographolide potassium succinate.

2. The human immunoglobulin E antagonist according to claim 1, characterized in that The andrographolide potassium succinate is obtained by the following screening method: (a) Establishment of an FDA-approved drug library: A library containing at least 3067 compound drugs was customized, each drug was dissolved in DMSO at a concentration of 1 mg / ml and stored; (b) preparing human immunoglobulin E target: customizing a human immunoglobulin E protein target with a purity of at least 95%, and gradiently diluting it into 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 is assembled into a microfluidic chip and installed in an appropriate high-throughput screening analyzer. After the chip quality is verified using a positive control sample, the human immunoglobulin E target is injected in sequence according to a concentration gradient. The interaction with the human immunoglobulin E target is detected, and the collected signal data is analyzed using data analysis software, and the results are ranked according to the molecular affinity; (d) Candidate drug screening: From the results obtained in step (c), a compound drug having a significant affinity reduction effect on the human immunoglobulin E target is identified, among which andrographolide potassium succinate is screened as a candidate drug with potential antagonistic effect; (e) Drug molecular level verification: A human immunoglobulin E detection kit was used, with known human immunoglobulin E monoclonal antibody drugs as positive controls, to evaluate the antagonistic effect of andrographolide potassium succinate on human immunoglobulin E. The antagonistic activity of andrographolide potassium succinate was verified by calculating the antagonism rate.

3. The human immunoglobulin E antagonist according to claim 2, characterized in that: The andrographolide potassium succinate is confirmed to have a significant affinity-reducing effect in step (d), and shows an antagonism rate at least equivalent to that of the positive control drug through ELISA detection in step (e).

4. The human immunoglobulin E antagonist according to claim 3, characterized in that: The FDA-approved drug library may contain more types of compound drugs to improve the diversity and accuracy of screening. The purity of the human immunoglobulin E target can be determined by SDS-PAGE or other methods to ensure that it meets the screening requirements.

5. The human immunoglobulin E antagonist according to claim 4, characterized in that: The high-throughput screening analyzer may be a PlexArray HT SPRi microarray analyzer or other instruments suitable for molecular screening; In the drug molecule level verification step, the antagonism rate is calculated as follows: antagonism rate = 100% - (detection concentration / initial concentration) x 100%.

6. The human immunoglobulin E antagonist according to claim 5, characterized in that: In step (e), the IL-5R monoclonal antibody drug omalizumab is used as a positive control, and the IL-5R amino acid sequence is shown in SEQ ID NO:

1.

7. An antagonist for improving the side effects of IgE antibody drug therapy, characterized in that: The main component of the antagonist is andrographolide potassium succinate.

8. Use of andrographolide potassium succinate as a human immunoglobulin E antagonist.

9. An antiallergic pharmaceutical composition containing andrographolide potassium succinate.

10. A method for treating or preventing allergic diseases using andrographolide potassium succinate.

Images