Lipid metabolite biomarker for predicting asthma treatment response to biological agent and use thereof

Abstract

The present invention relates to a biomarker for predicting an asthma treatment response to a biological agent and a use thereof, and specifically to a method for predicting an asthma treatment effect

Description

Lipid metabolite biomarkers and their use for predicting asthma treatment responsiveness to biological agents

[0001] This invention claims priority to Republic of Korea Patent Application No. 10-2023-0066126, filed May 23, 2023, the entire disclosure of which is incorporated herein by reference.

[0002] The present invention relates to a biomarker and its use for predicting asthma treatment responsiveness to a biological agent, and more particularly, to a 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 asthma treatment effect of a biological agent using LTE4.

[0003] Asthma is a disease characterized by bronchial hyperresponsiveness to various stimuli, causing chronic airway inflammation and clinical symptoms such as wheezing, dyspnea, and coughing caused by widespread airway narrowing, which can be improved naturally or reversibly with treatment. In general, asthma is recognized as a chronic inflammatory disease that occurs when inflammatory cells proliferate, differentiate, and are activated by interleukin-4, 5, and 13 produced by TH2 type immune cells, and 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.), which play an important role in the process of strong bronchoconstriction (Korean Patent Publication No. 10-2021-0119131).

[0004] Because this type of asthma is typically poorly controlled and prone to recurrent exacerbations, systemic steroids are frequently used to treat exacerbated asthma. However, due to the significant side effects of frequent steroid use, biologic agents have recently emerged as new, effective treatments for asthma. Examples of biologic agents include omalizumab, mepolizumab, reslizumab, benralizumab, and dupilumab.

[0005] However, these biologics are often not covered by insurance. Even when covered, they are often used only after the first-line treatment has been completed and the disease has progressed significantly, making them cost-prohibitive. Furthermore, because biologics target different substances depending on their type, their effectiveness varies depending on the specific patient. Therefore, when attempting to treat asthma with biologics, quickly predicting therapeutic efficacy and deciding whether to proceed with treatment is crucial for reducing both financial burden and patient suffering.

[0006] Therefore, if there is a method to predict the therapeutic effect of biological agents in asthma patients, it is expected that not only will the treatment cost and suffering of severe asthma patients be significantly reduced, but the treatment effect will also be significantly increased by selecting the biological agent that is right for the individual.

[0007] The present invention has been devised to solve the problems of the prior art as described above, and a) 2,3-Dinor-11β contained in a biological sample obtained from an asthma patient administered a biological agent 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-dinor-8-Iso-PGF 2α, 8,12-Iso-IPF 2α -VI, and LTE4; and b) classifying the patient as exhibiting a therapeutic response to the biological agent when the measured concentration of the eicosanoid is increased compared to before the treatment. The present invention provides a method for providing information on predicting the therapeutic response to a biological agent in an asthma patient.

[0008] However, the technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by a person having ordinary skill in the technical field to which the present invention belongs from the description below.

[0009] The present invention relates to a) a biological sample obtained from an asthma patient administered with a biological agent, wherein 2,3-Dinor-11β is 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 LTE4; and b) classifying the patient as exhibiting a therapeutic response to the biological agent when the measured concentration of the eicosanoid is increased compared to before the treatment. A method for providing information on predicting the therapeutic response to a biological agent in an asthma patient is provided.

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

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

[0012] In another specific embodiment of the present invention, the biological agent is preferably omalizumab, mepolizumab, reslizumab, benralizumab, or dupilumab, but is not limited thereto as long as it is a biological agent used as a treatment for asthma.

[0013] In addition, 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 -VI, and a composition for predicting the therapeutic response of a biological agent in an asthma patient, comprising as an active ingredient an agent measuring the concentration of one or more eicosanoids selected from the group consisting of LTE4.

[0014] In addition, the present invention provides a kit for predicting the therapeutic response of an asthma patient to a biological agent, comprising a composition for predicting the therapeutic response of an asthma patient to a biological agent as an active ingredient.

[0015] In addition, the present invention provides a diagnostic device for predicting the therapeutic response of an asthma patient to a biological agent, which comprises a composition for predicting the therapeutic response of an asthma patient to a biological agent as an active ingredient.

[0016] The present invention also provides a method for treating asthma, comprising 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 2α-VI, and LTE4; and d) when the measured concentration of the eicosanoid increases compared to before treatment, a method for treating asthma is provided, comprising the step of re-administering the biological agent of step a) to the patient.

[0017] In addition, the present invention provides a step of a) administering a biological agent to an asthma patient; b) obtaining a biological sample from the asthma patient; c) obtaining a biological sample from the asthma patient; and c) obtaining a biological sample from the 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, comprising as an active ingredient a composition for predicting the therapeutic response of a patient with asthma to a biological agent, the composition comprising the step of measuring the concentration of an eicosanoid contained in the biological sample; and d) when the concentration of the measured eicosanoid increases compared to before the treatment, the composition provides a method for treating asthma, the method comprising the step of re-administering the biological agent of step a) to the patient.

[0018] The method of providing information on predicting the therapeutic response of an asthma patient to a biological agent according to the present invention can identify a biological agent that exhibits a therapeutic effect suitable for each asthma patient, thereby significantly reducing the treatment cost and suffering of asthma patients, and can also significantly increase the therapeutic effect by selecting a biological agent that is suitable for the patient.

[0019] Figure 1 is a drawing showing the results of measuring the concentration of eicosanoids according to the presence or absence of a treatment response according to one embodiment of the present invention.

[0020] Figure 2 is a diagram showing the results of measuring the concentration of eicosanoids according to the treatment period according to one embodiment of the present invention.

[0021] A method of providing information on predicting therapeutic response to a biological agent of an asthma patient of the present invention comprises: 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 as biomarkers, thereby identifying effective biological agents that exhibit therapeutic effects in individual asthma patients.

[0022]

[0023] In this specification, “asthma” is a disease of the “bronchi,” the passages connecting to the lungs, which is a disease in which the bronchi are severely narrowed due to inflammation of the bronchi, causing repeated symptoms such as coughing, wheezing, shortness of breath, and chest tightness. Asthma includes, but is not limited to, allergic asthma or non-allergic asthma, severe asthma or uncontrolled asthma, adult asthma or childhood asthma. The “severe asthma” or “uncontrolled asthma” generally corresponds to, but is not limited to, severe asthma as defined in the American Thoracic Society and the European Respiratory Society (ATS / ERS) guidelines. The “uncontrolled asthma” refers to asthma in which the asthma condition is not controlled despite appropriate step-by-step asthma treatment, such as inhaled steroids appropriate for each asthma severity, and may include one or more of the following conditions, but is not limited to:

[0024] 1) Low symptom control: Partial control status according to the Korean Asthma Management Guidelines, or a score of less than 20 on the asthma control test.

[0025] 2) Frequent asthma exacerbations: Asthma exacerbations requiring systemic steroids for more than 3 days more than twice a year

[0026] 3) Severe exacerbation: When asthma worsens and hospitalization or mechanical ventilation is required.

[0027] 4) Airflow limitation: FEV1 < 80% after bronchodilator use.

[0028] In this specification, “biological agent” refers to a general term for a medical preparation made from a living organism, examples of which include serum, antigen, antibody, vitamin, hormone, cytokine, cell, plasma, etc., and preferably includes omalizumab, mepolizumab, reslizumab, benralizumab, dupilumab, etc., but is not limited thereto if it is a biological agent that can be used as a treatment for asthma. The dosage of the biological agent of the present invention may vary depending on various factors including the activity of the specific compound used, age, body weight, general health, sex, diet, administration time, administration route, excretion rate, drug combination, and the severity of the specific disease to be prevented or treated, and may be appropriately selected by those skilled in the art, although it may vary depending on the patient's condition, body weight, degree of disease, drug form, administration route, and period, and may be administered at 0.0001 to 500 mg / kg or 0.001 to 500 mg / kg per day. The administration may be administered once a day or divided into several times. The above dosage does not limit the scope of the present invention in any way.

[0029] In this specification, "biological sample" means any sample that can be used to determine the concentration of eicosanoids in the body, and preferably may be blood, plasma, serum, bone marrow, tissue, cells, saliva, sputum, peritoneal fluid, hair, urine, feces, cerebrospinal fluid, various secretions, etc., but is not limited thereto as long as it is a sample that can be obtained by a non-invasive method and contains eicosanoids. The sample may be pretreated by methods such as homogenization, filtration, distillation, extraction, concentration, inactivation of interfering components, and addition of reagents before use for detection or diagnosis.

[0030] As used herein, “eicosanoid” refers to an endogenous compound produced by the action of a cyclooxygenase enzyme system or a lipooxygenase enzyme system on arachadonic acid, an unsaturated fatty acid with 20 carbons, and exhibits various 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 thereto. Methods for measuring the eicosanoids include, but are not limited to, mass spectrometry, gas chromatography, liquid chromatography, mass spectrometry, column, mass spectrometry, ELISA, NMR, UV-Vis, HPLC, UPLC, and enzymatic-based colorimetric methods.

[0031] In this specification, “response” can be defined as “responsive” to a biological agent if asthma symptoms are prevented, improved, or treated as a result of biological agent treatment in an asthma patient. Prevention, improvement, or treatment of asthma can be measured in various known ways. The “prevention” refers to a broad concept of blocking the occurrence of asthma, and preferably includes both primary prevention, which prevents the occurrence in advance, and secondary prevention, which detects the occurrence early and treats it in a timely manner, but is not limited to the process and / or activity for dealing with the occurrence of asthma. The “treatment” refers to a broad concept of dealing with the occurrence of asthma, and is not limited to the process and / or activity for treating, curing, alleviating, or reducing asthma. The “improvement” refers to an act of at least reducing a parameter related to the condition being treated, for example, the severity of symptoms.

[0032] In this specification, the “method for providing information”, that is, the “method for providing information”, refers to a method for providing information on the asthma treatment effect of a biological agent, and means a method for obtaining or predicting information on the possibility that asthma symptoms will be prevented, improved, or treated when the biological agent is administered to an asthma patient. More specifically, it is a concept that includes all of selecting a biological agent suitable for an asthma patient, determining the susceptibility of an asthma patient to a biological agent, determining the prognosis of an asthma patient, or therametrics (e.g., monitoring the condition of an asthma patient to provide information on the treatment efficacy).

[0033] In this specification, "kit" means 2,3-Dinor-11β contained in a biological sample. 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α -VI, and / or LTE4, and means a screening device that can predict the treatment response of an asthma patient to a biological agent, and preferably, it can be in the form of including an antibody, compound, etc. that binds to the eicoside, but 2,3-Dinor-11β from a biological sample isolated from an asthma patient. 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 form can measure the amount of -VI and / or LTE4.

[0034] In this specification, "diagnosis instrument" means a device capable of measuring therapeutic response to biological agents in vitro based on biological samples generated from the human body, such as blood, saliva, urine, etc., and preferably an inlet for adding biological samples, 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, and may include a body capable of measuring the amount, an output unit showing the measured result, etc., but 2,3-Dinor-11β contained in a 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 the type of equipment that can measure the amount of -VI and / or LTE4.

[0035]

[0036] Hereinafter, preferred examples are presented to aid in understanding the present invention. However, the following examples are provided solely to facilitate a better understanding of the present invention, and the scope of the present invention is not limited by the following examples.

[0037]

[0038] [Example]

[0039] Example 1: Quantitative analysis of metabolites in asthma patients

[0040] To identify biomarkers predicting the efficacy of biologic agents in asthma treatment, urine samples were collected from 83 patients (PRISM study cohort, hosted by the Ministry of Science and Technology) who visited for asthma treatment during each of three treatment courses. Each patient was treated with either biologic agents or conventional treatment. The amounts of eicosanoids, lipid metabolites, in urine samples were determined. Eicosanoids were analyzed using solid-phase extraction (SPE) pretreatment. Specifically, 3 cc / 60 mg Evolute Express ABN SPE cartridges were used for analysis on an Extrahera automated instrument. Eicosanoids from the PG metabolites, thromboxanes, and isoprostanes were analyzed using 300 μL of urine. Additionally, 2,3-dinor-TXB2, a tautomer, was analyzed using a derivatization method. LTE4, a CysLTs, was analyzed using 1 mL of urine. A Waters Acquity UPLC system was used as the liquid chromatography instrument, and an AB Sciex Triple Quadrupole 5500 mass spectrometer was used. An ACQUITY UPLC®HSS T3 (100 × 2.1 mm id, 1.8 μm particle size) column was used for separation.

[0041] Because a single spot urine sample can affect the concentration of metabolites in urine depending on body water content, the concentration was adjusted using the specific gravity of water, and the calculation formula is as follows. The results are shown in Table 1.

[0042] [Formula 1]

[0043] Concentration corrected = Concentrationmeasured X (1.020 - 1) / (Specific gravity - 1)

[0044] After preprocessing, the quantitative analysis of eicosanoids was performed to determine the quantitative range of eicosanoids. Specifically, the Multiple Reaction Monitoring (MRM) transition values ​​and retention times (RT) for 10 types of eicosides were measured. The highly sensitive Sciex Triple Quad 5500 was used for the measurements to lower the lower limit of quantitation (LOQ). The quantitative range of a total of 10 eicosanoids was confirmed. The results are shown in Table 2. Samples for quality control (QC) were then prepared using the quantitative range.

[0045] CompoundMRMRTMethod 12,3,-dinor-11β-PGF2α325.3 → 145.15.49Tetranor PGEM327.1 → 143.01.9011-Dehydro-TXB2367.5 → 161.07.698,12-Iso-iPF 2α -VI353.3 → 115.08.642,3-Dinor-8-iso-PGF2α325.1 → 237.25.13 PGF2α 353.2 → 309.17.498-Iso-PGF2α353.3 → 193.16.74 PGE2 351.5 → 271.07.69PGF 2α -d4357.1 → 197.27.48PGE2-d9360.0 → 280.37.61Tetranor PGEM-d6333.1 → 315.11.8611-Dehydro-TXB2-d4371.3 → 165.07.658-Iso-PGF 2α -d4357.1 → 251.26.70Method 22,3-Dinor-TXB2370.0 → 155.03.43TXB2-d4402.1 → 173.03.90Method 3 LTE4438.1 → 333.02.34LTE4-d5443.1 → 338.02.31

[0046] (from-wi: ng / mL)2,3-Dinor- 11β-PGF2α Tetranor PGEM 11-Dehydro-TXB2 8,12-Iso-IPF 2α -VI2,3-denarius-8-Jesus- PGF2α PGF2α 8-Jesus- PGF2α PGE2 2.3-dinar-TXB2 LTE4 STD10.30720.8960.10.35840.30720.23040.0640.07680.050.01STD20.61441.7920.20.71680.61440.4 6080.1280.15360.100.0STD31.22883.5840.41.43361.22880.92160.2560.30720.50.0STD42.45767.168 0.82.86722.45761.84320.5120.614420STD56.14417.9227.1686.1444.6081.281.53650STD615.3644.8 517.9215.3611.523.23.84202STD719.211212.544.838.428.889.6804STD89628031.25112967220241008

[0047] The urine samples of patients treated with biologic agents such as mepolizumab, dupilumab, reslizumab, and / or omalizumab were used to analyze the differences according to treatment response, compared to the urine samples of patients treated with conventional methods such as steroids and inhalers. The treatment response was analyzed by dividing patients into responders (R) and non-responders (NR) based on whether the prebronchodilator FEV1 increased by 100 mL or more or 10% or more after starting treatment with biologic agents, and the results were expressed as the mean ± standard deviation. Statistical significance was confirmed by the Mann Whitney U test, and statistical significance was determined if P < 0.05. The results are shown in Fig. 1.

[0048] 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 -VI, and LTE4 concentrations were significantly increased when the responder was present. These results confirmed that the asthma treatment efficacy of biological agents can be predicted using the six eicosanoids.

[0049]

[0050] Example 2: Quantitative analysis of eicosanoids according to treatment period

[0051] To determine the concentration of eicosanoids according to the treatment period, the concentration of eicosanoids was measured using urine samples taken before treatment (visit 1), 1 month after treatment (visit 2), and 6 months after treatment (visit 3) using the same method as in Example 1. The results are shown in Fig. 2.

[0052] As shown in Fig. 2, PGF 2α It was confirmed that the concentration of LTE4 significantly changed depending on the treatment time.

[0053]

[0054] The foregoing description of the present invention is provided for illustrative purposes only. Those skilled in the art will readily appreciate that the present invention can be readily modified into other specific forms without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

[0055] A method for providing information on predicting the therapeutic response of an asthma patient to a biological agent of the present invention comprises selecting 2,3-Dinor-11β among various eicosanoids. 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α-VI, and LTE4 can predict the responsiveness to biological agents with high accuracy, and through this, a patient group that responds to biological agents can be selected and treated. Therefore, compared to the existing patient selection system, that is, a system that decides to administer biological agents only to patients with long-term asthma exacerbation and determines whether to proceed with or discontinue treatment after confirming the effect for more than 6 months after administration, not only can the patient's suffering and economic burden be reduced, but also the economic cost of health insurance can be significantly reduced. Therefore, by using the method for providing information on predicting the therapeutic responsiveness of an asthma patient to a biological agent of the present invention, the composition for predicting the therapeutic responsiveness, the kit thereof, the diagnostic device thereof, etc., a patient group that responds to the treatment when the biological agent is administered can be easily selected. Therefore, the method for providing information on predicting the therapeutic responsiveness of an asthma patient to a biological agent of the present invention can quickly and accurately predict the therapeutic effect using a biological sample obtained by a non-invasive method, and thus a patient group that responds to the biological agent can be easily selected and a decision made on whether to proceed with the treatment.

Claims

1. a) 2,3-Dinor-11β contained in a biological sample obtained from an asthma patient administered a biological agent 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α -VI, and a step of measuring the concentration of one or more eicosanoids selected from the group consisting of LTE4; and b) A method for providing information on predicting treatment responsiveness to a biological agent in an asthma patient, comprising the step of classifying the patient as exhibiting a treatment response to the biological agent when the concentration of the measured eicosanoid is increased compared to before treatment.

2. In paragraph 1, A method, characterized in that the biological sample is urine, saliva, blood, plasma or serum.

3. In paragraph 1, A method characterized in that the above asthma is asthma for Koreans.

4. In paragraph 1, A method, characterized in that the biological agent is at least one selected from the group consisting of omalizumab, mepolizumab, reslizumab, benralizumab, and dupilumab. 5.2,3-Dinor-11β 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α A composition for predicting therapeutic response to a biological agent in an asthma patient, comprising as an active ingredient an agent for measuring the concentration of one or more eicosanoids selected from the group consisting of -VI, and LTE4.

6. A kit for predicting therapeutic response to a biological agent in an asthma patient, comprising a composition for predicting therapeutic response to a biological agent in an asthma patient under paragraph 5 as an active ingredient.

7. A diagnostic device for predicting the therapeutic response of an asthma patient to a biological agent, comprising a composition for predicting the therapeutic response of an asthma patient to a biological agent in accordance with Article 5 as an active ingredient. 8.a) Step of administering a biological agent to an asthma patient; b) obtaining a biological sample from the asthma patient; c) 2,3-Dinor-11β contained in the above biological sample 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α -VI, and a step of measuring the concentration of one or more eicosanoids selected from the group consisting of LTE4; and d) A method for treating asthma, comprising the step of re-administering the biological agent of step a) to the patient when the concentration of the measured eicosanoid increases compared to before treatment. 9.a) Step of administering a biological agent to an asthma patient; b) obtaining a biological sample from the asthma patient; c) 2,3-Dinor-11β contained in the biological sample using a composition for predicting the therapeutic response to a biological agent of an asthma patient in Article 5 2α , 11-Dehydro-TXB2, PGF 2α , 2,3-dinor-8-Iso-PGF 2α , 8,12-Iso-IPF 2α -VI, and a step of measuring the concentration of one or more eicosanoids selected from the group consisting of LTE4; and d) A method for treating asthma, comprising the step of re-administering the biological agent of step a) to the patient when the concentration of the measured eicosanoid increases compared to before treatment.

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