Lipid metabolite biomarkers for predicting asthma treatment response to biological agents and their applications

Abstract

The present invention relates to a biomarker for predicting asthma treatment response to biological agents and its applications, more specifically, to 2,3-Dinor-11β 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-Dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α -VI, or a method for predicting the therapeutic effect of biological agents on asthma using LTE4, etc.

Description

[Technical Field]

[0001] This invention claims priority under Republic of Korea Patent Application No. 10-2023-0066126, filed on 23 May 2023, and all contents disclosed in the specification and drawings of said application are incorporated herein by reference.

[0002] The present invention relates to a biomarker for predicting the response of asthma to biological agents and its applications, and more specifically, to the lipid metabolite 2,3-Dinor-11β 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-Dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α -VI, or a method for predicting the therapeutic effect of biological agents on asthma using LTE4, etc. [Background technology]

[0003] Asthma is a disease characterized by bronchial hyperresponsiveness to various stimuli, leading to chronic airway inflammation and clinical symptoms such as wheezing, shortness of breath, and cough, which result from widespread narrowing of the airways. These symptoms can improve spontaneously or reversibly with treatment. Generally, asthma is recognized as a chronic inflammatory disease in which inflammatory cells proliferate, differentiate, and are activated by interleukins-4, 5, and 13 produced by TH2-type immune cells, and then migrate and infiltrate the airways and surrounding tissues. In this case, activated inflammatory cells such as eosinophils, mast cells, and alveolar macrophages secrete various inflammatory mediators (cysteine ​​leukotrienes, prostaglandins, etc.) and exert a strong bronchoconstrictive effect, playing a crucial role in the process (Republic of Korea Published Patent 10-2021-0119131).

[0004] This type of asthma is generally not well controlled and tends to worsen repeatedly, leading to frequent use of systemic corticosteroids to treat exacerbated asthma. However, frequent use of corticosteroids has significant side effects, so recently, biological agents have been attracting attention as new formulations for effectively treating asthma. Examples of biological agents currently in use include omalizumab, mepolizumab, reslizumab, benralizumab, and dupilumab.

[0005] However, these biological agents are mostly not covered by insurance, and even when they are, they are usually only applied when the disease has progressed for a long period of time without treatment after completing all stages of primary treatment, making them financially difficult to use. Furthermore, since different types of biological agents target different substances, their effects vary depending on which drug is used for which patient. Therefore, when attempting to treat asthma with biological agents, it is crucial to quickly predict the treatment effect and determine whether treatment is feasible in order to reduce the financial burden and patient suffering.

[0006] Therefore, if there were a method to predict the therapeutic effect of biological agents on asthma patients, it is expected that not only could the treatment costs and suffering of severe asthma patients be significantly reduced, but the therapeutic effect could also be significantly improved by selecting a biological agent that is right for the individual. [Overview of the project] [Problems that the invention aims to solve]

[0007] The present invention was devised to solve the above-mentioned problems of the prior art, and a) 2,3-Dinor-11β contained in biological samples obtained from asthma patients administered biological agents. 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-Dinor-8-Iso-PGF 2α, 8,12-Iso-IPF 2α -VI, and measuring the concentration of one or more eicosanoids selected from the group consisting of LTE4; and b) classifying a patient as showing a therapeutic effect on the biological agent when the concentration of the measured eicosanoid has increased compared to before treatment. The purpose is to provide a method for predicting the therapeutic responsiveness of a biological agent in asthmatic patients, etc.

[0008] However, the technical problems to be achieved by the present invention are not limited to the problems mentioned above, and other problems not mentioned should be clearly understood by those with ordinary knowledge in the technical field to which the present invention belongs from the following description.

Means for Solving the Problems

[0009] The present invention provides a method for predicting the therapeutic responsiveness of a biological agent in asthmatic patients, including: a) measuring the concentration of one or more eicosanoids selected from the group consisting of 2,3-Dinor-11β, 11-Dehydro-TXB2, PGF, 2,3-Dinor-8-Iso-PGF, 8,12-Iso-IPF-VI, and LTE4 contained in a biological sample obtained from an asthmatic patient to whom a biological agent has been administered; and b) classifying a patient as showing a therapeutic effect on the biological agent when the concentration of the measured eicosanoid has increased compared to before treatment. 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-Dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α -VI, and measuring the concentration of one or more eicosanoids selected from the group consisting of LTE4; and b) classifying a patient as showing a therapeutic effect on the biological agent when the concentration of the measured eicosanoid has increased compared to before treatment.

[0010] In one specific example of the present invention, the biological sample is preferably urine, saliva, blood, plasma, serum, but is not limited thereto as long as it is a sample that can be obtained non-invasively.

[0011] In another specific example of the present invention, the asthma is preferably asthma in Koreans, but is not limited thereto.

[0012] In yet another specific example of the invention, the biological agent is preferably omalizumab, mepolizumab, reslizumab, benralizumab, or dupilumab, but is not limited to any biological agent used as a treatment for asthma.

[0013] Furthermore, the present invention relates to 2,3-Dinor-11β 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-Dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α The present invention provides a composition for predicting the therapeutic response to biological agents in asthma patients, comprising as an active ingredient a formulation that measures the concentration of one or more eicosanoids selected from the group consisting of -VI and LTE4.

[0014] Furthermore, the present invention provides a kit for predicting the therapeutic response of asthma patients to biological agents, comprising the aforementioned composition for predicting the therapeutic response of asthma patients to biological agents as an active ingredient.

[0015] Furthermore, the present invention provides a diagnostic device for predicting the therapeutic response of asthma patients to biological agents, comprising a composition for predicting the therapeutic response of asthma patients to biological agents as an active ingredient.

[0016] Furthermore, the present invention comprises the steps of: a) administering a biological agent to an asthma patient; b) obtaining a biological sample from the asthma patient; and c) obtaining 2,3-Dinor-11β contained in the biological sample. 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-Dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2αThe present invention provides a method for treating asthma, comprising the steps of: - measuring the concentration of one or more eicosanoids selected from the group consisting of VI and LTE4; and d) administering the biological agent from step a) again to the patient when the measured concentration of the eicosanoid has increased compared to before treatment.

[0017] Furthermore, the present invention relates to a) the step of administering a biological agent to an asthma patient; b) the step of obtaining a biological sample from the asthma patient; and c) the 2,3-Dinor-11β 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-Dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α The present invention provides a method for treating asthma, comprising the steps of: (a) measuring the concentration of eicosanoids contained in a biological sample using a composition for predicting therapeutic response to a biological agent in an asthma patient, which contains as an active ingredient one or more eicosanoids selected from the group consisting of VI and LTE4; and (d) administering the biological agent from step (a) again to the patient when the measured concentration of eicosanoids has increased compared to before treatment. [Effects of the Invention]

[0018] The method for providing information on predicting the therapeutic response to biological agents in asthma patients according to the present invention allows for the identification of biological agents that are suitable for individual asthma patients and that show therapeutic effects. This not only significantly reduces the treatment costs and suffering of asthma patients but also significantly improves the therapeutic effect by allowing them to select a biological agent that is right for them. [Brief explanation of the drawing]

[0019] [Figure 1] Figure 1 shows the results of measuring the concentration of eicosanoids with and without a therapeutic response according to one embodiment of the present invention. [Figure 2] Figure 2 shows the results of measuring the concentration of eicosanoids during the treatment period according to one embodiment of the present invention. [Best Mode for Carrying Out the Invention]

[0020] The present invention provides information regarding the prediction of therapeutic response to biological agents in asthma patients, and is a method for providing information regarding 2,3-Dinor-11β 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-Dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α -VI and / or LTE4 are used as biomarkers to identify effective biological agents that show therapeutic effects in individual asthma patients.

[0021] In this specification, "asthma" refers to a disease of the "bronchi," the airways leading to the lungs, characterized by severe narrowing of the bronchi due to inflammation, resulting in recurrent cough, wheezing, shortness of breath, and chest tightness. Asthma includes, but is not limited to, allergic or non-allergic asthma, severe or poorly controlled asthma, adult or childhood asthma. "Severe asthma" or "poorly controlled asthma" generally corresponds to the American Thoracic Society and European Respiratory Society (ATS / ERS) guidelines, but is not limited to these. "Poorly controlled asthma" means asthma whose condition is not regulated despite appropriate, stepwise asthma treatment, such as inhaled corticosteroids, commensurate with the severity of each type of asthma, and may include, but is not limited to, one or more of the following conditions: 1) Low symptom control: Partial control status according to the Korean Asthma Treatment Guidelines, or less than 20 points on the Asthma Control Test. 2) Frequent asthma exacerbations: Asthma exacerbations requiring systemic steroids for 3 days or more occur at least twice a year. 3) Severe exacerbation: Asthma worsens to the point where hospitalization or mechanical ventilation is required. 4) Airflow restriction: FEV1 < 80% after use of bronchodilators.

[0022] In this specification, "biological agent" refers collectively to medical preparations made from living organisms, and examples include serum, antigens, antibodies, vitamins, hormones, cytokines, cells, plasma, etc. Preferably, omalizumab, mepolizumab, reslizumab, benralizumab, dupilumab, etc., but is not limited to these as long as it is a biological preparation that can be used as a treatment for asthma. The dosage of the biological preparation of the present invention can vary considerably depending on various factors including the activity of the specific compound used, age, weight, general health, sex, mental state, administration time, route of administration, elimination rate, drug formulation, and the severity of the specific disease being prevented or treated, and will vary depending on the patient's condition, weight, disease stage, drug form, route of administration, and duration, but can be appropriately selected by those skilled in the art and can be administered at a dose of 0.0001 to 500 mg / kg or 0.001 to 500 mg / kg per day. The drug may be administered once a day or in several divided doses. The aforementioned dosage does not limit the scope of the present invention in any way.

[0023] In this specification, "biological sample" means any sample from which the concentration of eicosanoids in the body can be confirmed, preferably including blood, plasma, serum, bone marrow, tissue, cells, saliva, sputum, peritoneal fluid, hair, urine, feces, cerebrospinal fluid, and various secretions, but not limited to any sample that contains eicosanoids and can be obtained by a non-invasive method. The sample may be pretreated before use for detection or diagnosis by methods such as homogenization, filtration, distillation, extraction, concentration, inactivation of interfering components, and addition of reagents.

[0024] In this specification, "eicosanoid" refers to endogenous compounds produced by the action of cyclooxygenase or lipoxygenase enzyme systems on arachidonic acid, a 20-carbon unsaturated fatty acid, and exhibits diverse physiological activities. Examples of eicosanoids include, but are not limited to, prostaglandins, prostacyclins, thromboxanes, leukotrienes, and PUFAs, or other metabolites formed by the enzymatic or non-enzymatic addition of oxygen to these. Methods for measuring eicosanoids include, but are not limited to, mass spectrometers, gas chromatography, liquid chromatography, mass spectrometers, columns, mass spectrometry, ELISA, NMR, UV-Vis, HPLC, UPLC, and enzymatic colorimetric methods.

[0025] In this specification, “response” can be defined as “response” to a biological agent if, as a result of treatment with the biological agent, the asthma symptoms are prevented, improved, or treated in the patient with asthma. Prevention, improvement, or treatment of asthma can be measured in a variety of known ways. “Prevention” is a broad concept of blocking the onset of asthma, preferably including primary prevention, which prevents onset in advance, and secondary prevention, which detects onset early and treats it in a timely manner, but is not limited to, any process and / or activity that addresses asthma before its onset. “Treatment” is a broad concept of addressing the onset of asthma, and is not limited to any process and / or activity that causes the treatment, cure, alleviation, reduction, etc. of asthma. “Improvement” means an action that at least reduces the severity of symptoms, for example, a parameter related to the condition being treated.

[0026] In this specification, “method for providing information” means a method for providing information regarding the therapeutic effect of a biological agent on asthma, which involves obtaining or predicting information about the likelihood that asthma symptoms will be prevented, improved, or treated when the biological agent is administered to an asthma patient. More specifically, it encompasses the selection of a biological agent suitable for an asthma patient, determining the asthma patient’s susceptibility to the biological agent, determining the asthma patient’s prognosis, or the use of theametrics (e.g., monitoring the asthma patient’s condition to provide information about therapeutic efficacy).

[0027] In this specification, "kit" refers to the 2,3-Dinor-11β contained in the biological sample. 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-Dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α -VI and / or LTE4 amounts can be measured to predict the therapeutic response of asthma patients to biological agents, and preferably, this may be in the form of antibodies, compounds, etc. that bind to the eicosanoid, but also from biological samples isolated from asthma patients. 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-Dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α - There are no restrictions as long as the amount of VI and / or LTE4 can be measured.

[0028] In this specification, "diagnosis instrument" means equipment capable of measuring the therapeutic response to a biological agent outside the body based on a biological sample generated in the human body, such as blood, saliva, or urine, and preferably includes an inlet for adding the biological sample, and 2,3-Dinor-11β 2α, 11-Dehydro-TXB2, PGF 2α , 2,3-Dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α -The device may include a main unit capable of measuring the amount of VI and / or LTE4, an output unit that displays the measured results, etc., but which contains 2,3-Dinor-11β in the biological sample. 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-Dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α - There are no restrictions on equipment that can measure the amount of VI and / or LTE4.

[0029] The following are preferred embodiments to aid in understanding the present invention. However, the following embodiments are provided only to facilitate understanding of the present invention, and the scope of the present invention is not limited by these embodiments. [Examples]

[0030] Example 1: Quantitative analysis of metabolites in asthma patients To identify biomarkers for predicting the efficacy of biological agents in asthma treatment, urine samples were collected during three treatment cycles from 83 patients (a cohort of the Science and Technology Department's PRISM study) who visited the hospital for asthma treatment. Each patient underwent asthma treatment with either biological agents or existing conventional treatment methods. The amount of eicosanoids, which are lipid metabolites, in the urine samples was then examined. Eicosanoids were analyzed using a solid-phase extraction (SPE) pretreatment method. More specifically, analysis was performed using an Extrahera automated instrument with 3cc / 60mg Evolute Express ABN SPE cartridges. Eicosanoids from the PG metabolites, thromboxanes, and isoprostans series were analyzed using 300 μL of urine. Additionally, the tautomer 2,3-dinor-TXB2 was analyzed using an additional derivatization method. The CysLTs series LTE4 was analyzed using 1 mL of urine. For liquid chromatography, a Waters Acquity UPLC system was used, and for mass spectrometry, an AB Sciex Triple Quadrupole 5500 was employed. For separation, an ACQUITY UPLC® HSS T3 column (100 × 2.1 mm id, 1.8 μm particle size) was used.

[0031] Since the concentration of metabolites in a single urine sample can be affected by the body's water content, the concentration is corrected using the specific gravity relative to water, and the calculation formula is as follows. The results are shown in Table 1 below.

[0032] Concentration corrected =Concentration measured X(1.020-1) / (Specific gravity-1) (Equation 1)

[0033] After sample preparation, the quantitative range of eicosanoids was confirmed for quantitative analysis. More specifically, Multiple Reaction Monitoring (MRM) transition values ​​and retention time (RT) were measured for 10 types of eicosides. A highly sensitive Sciex Triple Quad 5500 was used during measurement to lower the minimum limit of quantification. The quantitative ranges of a total of 10 eicosanoids were confirmed. The results are shown in Table 2. Then, samples for quality control (QC) were prepared using the quantitative ranges.

[0034] [Table 1]

[0035] [Table 2]

[0036] We analyzed the differences in treatment response between patients treated with the biological agents mepolizumab, dupilumab, reslizumab, and / or omalizumab, and patients treated with conventional methods, i.e., steroids, inhalants, etc. Treatment response was determined by whether prebronchodilator FEV1 increased by 100 mL or more, or by 10% or more, after initiating treatment with the biological agent. Patients were divided into responders (R) and non-responders (NR), and the results are presented as mean ± standard deviation. Statistical significance was confirmed by the Mann-Whitney U test; P < 0.05 was considered statistically significant. The results are shown in Figure 1.

[0037] As shown in Figure 1, among eicosanoids, 2,3-Dinor-11β 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-Dinor-8-Iso-PGF 2α, 8,12-Iso-IPF 2α We confirmed that the concentrations of -VI and LTE4 significantly increased in responders. Through these results, we were able to confirm that the asthma treatment effect of biological agents can be predicted using the six eicosanoids mentioned above.

[0038] Example 2: Quantitative analysis of eicosanoids based on treatment period To confirm the concentration of eicosanoids over the treatment period, eicosanoid concentrations were measured using urine samples taken before treatment (visit 1), one month after treatment (visit 2), and six months after treatment (visit 3) using the same method as in Example 1. The results are shown in Figure 2.

[0039] As shown in Figure 2, PGF 2α We confirmed that the concentration of LTE4 changes significantly depending on the time of treatment.

[0040] The above description of the present invention is illustrative, and a person with ordinary skill in the art to which the present invention pertains will understand that it can be easily modified into other specific forms without altering the technical idea or essential features of the present invention. Therefore, the embodiments described above should be understood in all respects as illustrative and not limiting. [Industrial applicability]

[0041] The present invention provides information regarding the prediction of therapeutic response to biological agents in asthma patients, and among a variety of eicosanoids, 2,3-Dinor-11β 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-Dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α-VI and LTE4 can predict with high accuracy whether or not a patient will respond to a biological agent. Through this, a group of patients who will respond to the biological agent can be selected and treatment can be advanced accordingly. Compared to existing patient selection systems, which only administer biological agents to patients with long-term asthma exacerbations and then confirm the effect for more than six months before deciding whether to continue or discontinue treatment, this system can reduce not only patient suffering and financial burden, but also significantly reduce the financial costs of medical insurance. Therefore, by using the method for providing information on predicting the therapeutic response of asthma patients to biological agents, the therapeutic response prediction composition, the kit, the diagnostic equipment, etc. of the present invention, a group of patients who will respond to the treatment can be easily selected when administering a biological agent. Therefore, the method for providing information on predicting the therapeutic response of asthma patients to biological agents of the present invention can quickly and accurately predict the therapeutic effect using biological samples obtained by a non-invasive method, making it possible to easily select a group of patients who will respond to the biological agent and decide whether or not to proceed with treatment.

Claims

1. a) 2,3-Dinor-11β contained in biological samples obtained from asthma patients who received biological agents. ２α , 11-Dehydro-TXB ２ PGF ２α , 2,3-Dinor-8-Iso-PGF ２α ,8,12-Iso-IPF ２α -VI and LTE ４ A step of measuring the concentration of one or more eicosanoids selected from the group consisting of; and b) A method for providing information regarding the prediction of therapeutic response to a biological agent in an asthma patient, characterized by including the step of classifying the patient as one who shows a therapeutic effect to a biological agent when the measured concentration of the eicosanoid increases compared to before treatment.

2. The method according to claim 1, characterized in that the biological sample is urine, saliva, blood, plasma, or serum.

3. The method according to claim 1, characterized in that the asthma is asthma in Koreans.

4. The method according to claim 1, characterized in that the biological agent is one or more selected from the group consisting of omalizumab, mepolizumab, reslizumab, benralizumab, or dupilumab.

5. 2,3-Dinor-11β ２α 、11-Dehydro-TXB ２ 、PGF ２α 、2,3-Dinor-8-Iso-PGF ２α 、8,12-Iso-IPF ２α -VI、及びLTE ４ A composition for predicting the therapeutic responsiveness to a biological preparation for asthmatic patients, characterized by containing, as an active ingredient, a preparation for measuring the concentration of one or more eicosanoids selected from the group consisting of

6. A kit for predicting the therapeutic response of asthma patients to biological agents, characterized by comprising the composition for predicting the therapeutic response of asthma patients to biological agents as described in claim 5 as an active ingredient.

7. A diagnostic device for predicting the therapeutic response of asthma patients to biological agents, characterized by containing the composition for predicting the therapeutic response of asthma patients to biological agents as described in claim 5 as an active ingredient.

8. a) The stage of administering biological agents to asthma patients; b) The step of obtaining a biological sample from the asthma patient; c) 2,3-Dinor-11β contained in the biological sample ２α , 11-Dehydro-TXB ２ PGF ２α , 2,3-Dinor-8-Iso-PGF ２α ,8,12-Iso-IPF ２α -VI and LTE ４ A step of measuring the concentration of one or more eicosanoids selected from the group consisting of; and d) A method for treating asthma, characterized by comprising the step of administering the biological agent from step a) again to the patient when the measured concentration of the eicosanoid has increased compared to before treatment.

9. a) The stage of administering biological agents to asthma patients; b) The step of obtaining a biological sample from the asthma patient; c) Using the composition for predicting therapeutic response to a biological agent in an asthma patient according to claim 5, 2,3-Dinor-11β contained in the biological sample ２α , 11-Dehydro-TXB ２ PGF ２α , 2,3-Dinor-8-Iso-PGF ２α ,8,12-Iso-IPF ２α -VI and LTE ４ A step of measuring the concentration of one or more eicosanoids selected from the group consisting of; and d) A method for treating asthma, characterized by comprising the step of administering the biological agent from step a) again to the patient when the measured concentration of the eicosanoid has increased compared to before treatment.

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