EDN biomarker for predicting therapeutic responsiveness to antibody therapeutic agent for asthma

Abstract

The present invention relates to a composition and method capable of effectively predicting the therapeutic responsiveness of a subject to an antibody therapeutic agent for asthma, using blood EDN levels or urine EDN levels. In particular, the present invention can significantly increase the efficiency of predicting therapeutic responsiveness by configuring, as appropriate parameters, blood and urine EDN levels together with the number of blood eosinophils that were conventionally used to predict therapeutic responsiveness to antibody therapeutic agents for asthma. Therefore, the present invention can be usefully utilized for predicting the therapeutic responsiveness of a subject to various antibody therapeutic agents for asthma, such as mepolyizumab, resluzumab, and dupilumab.

Description

EDN biomarkers for predicting treatment responsiveness to antibody therapies for asthma

[0001] [Cross-reference with related applications]

[0002] This application claims the benefit of priority based on Korean Patent Application No. 10-2024-0197172 dated December 26, 2024, and all contents disclosed in the document of said Korean patent application are incorporated herein as part of this specification.

[0003] [Technology Field]

[0004] The present invention relates to an EDN biomarker for predicting therapeutic responsiveness to an antibody therapeutic agent for asthma.

[0005] Asthma is a chronic inflammatory disease of the airways characterized by various airway obstructions, airway hyperresponsiveness, and airway inflammation. Since asthma is a chronic disease, the goal of asthma treatment is to achieve disease control, and in particular, to alleviate symptoms and reduce asthma exacerbations to improve the quality of life.

[0006] Meanwhile, eosinophils, a type of inflammatory granulocyte, are involved in the pathophysiology of asthma by playing a significant role in airway inflammation. Increased numbers of eosinophils have been reported in bronchoalveolar lavage fluid, sputum, and / or peripheral blood of asthmatic subjects. In particular, the accumulation of eosinophils in the airways can lead to inflammation and narrowing of the airways following degranulation and the release of inflammatory mediators. Eosinophilic inflammation is correlated with asthma severity, particularly the frequency of asthma exacerbations. Compared to the non-eosinophilic type, severe eosinophilic asthma has been reported to be characterized by severe exacerbations, concomitant sinusitis, peripheral airway involvement, airway remodeling, and decreased lung function. Conversely, severe non-eosinophilic asthma is characterized primarily by neutrophil inflammation and severe asthma symptoms. Classification and approaches based on these eosinophilic inflammation phenotypes are expected to aid in the selection of asthma treatments.

[0007] Most asthma can be well controlled, for example, by maintenance medications including corticosteroids, particularly inhaled corticosteroids and / or bronchodilators. However, eosinophilic asthma is known not to be well controlled by these maintenance medications.

[0008] Therefore, there is a continuing need to identify effective new biomarkers capable of predicting and determining which patients in various asthma patient groups will respond to treatment.

[0009] Under this technical background, the applicant has confirmed that using blood EDN levels or urine EDN levels can effectively predict the therapeutic responsiveness of a subject to an asthma antibody treatment. Therefore, it is expected that by using the present invention to predict a patient group with high therapeutic responsiveness before administering an antibody treatment to a subject, a treatment method suitable for each individual subject can be selected.

[0010] One objective of the present invention is to provide a composition for predicting the therapeutic responsiveness of a subject to an asthma antibody therapeutic agent.

[0011] Another objective of the present invention is to provide a kit for predicting the therapeutic responsiveness of a subject to an asthma antibody treatment.

[0012] Another objective of the present invention is to provide a method for predicting the therapeutic responsiveness of a subject to an asthma antibody treatment using a sample separated from the subject.

[0013] To achieve the above objective, one aspect of the present invention provides a composition for predicting the therapeutic responsiveness of a subject to an asthma antibody therapeutic agent, comprising as an active ingredient a preparation for measuring the expression level of mRNA of an EDN (Eosinophil-derived neurotoxin) gene or an EDN protein.

[0014] In addition, another aspect of the present invention provides a kit for predicting the therapeutic responsiveness of a subject to an asthma antibody therapeutic agent, comprising the above composition.

[0015] In addition, another aspect of the present invention provides a method for predicting the therapeutic responsiveness of a subject to an asthma antibody therapeutic agent, comprising the step of measuring the expression level of mRNA of the EDN (Eosinophil-derived neurotoxin) gene or the EDN protein in a sample separated from the subject.

[0016] The present invention has the effect of predicting a subject's therapeutic responsiveness to asthma antibody therapeutics using blood or urine EDN levels. In particular, the present invention can significantly improve the efficiency of predicting therapeutic responsiveness by configuring appropriate parameters with blood eosinophil counts, which were previously used to predict therapeutic responsiveness to asthma antibody therapeutics, using blood and urine EDN levels. Therefore, the present invention can be usefully utilized to predict a subject's therapeutic responsiveness to various asthma antibody therapeutics, such as mepolizumab, reslizumab, and dupilumab.

[0017] However, the effects of the present invention are not limited to those mentioned above, and other unmentioned effects will be clearly understood by those skilled in the art from the following description.

[0018] Figure 1 shows the blood eosinophil count, serum EDN levels, and urine EDN levels measured according to the duration of treatment in asthma patient groups treated with mepolizumab, reslizumab, or dupilumab.

[0019] The present invention will be described in detail below.

[0020]

[0021] 1. Composition for predicting the therapeutic responsiveness of a subject to an asthma antibody therapeutic agent

[0022] One aspect of the present invention provides a composition capable of predicting the therapeutic responsiveness of a subject to an asthma antibody therapeutic agent in asthma.

[0023] In the present invention, the composition comprises, as an active ingredient, a preparation for measuring the expression level of the mRNA of the EDN (Eosinophil-derived neurotoxin) gene or the EDN protein.

[0024] The above EDN (Eosinophil-derived neurotoxin) refers to a neurotoxin derived from eosinophils as a product of eosinophil degranulation. It can reduce the activity of ssRNA (single strand RNA) viruses through enzymatic activity and plays a role in attracting immune cells.

[0025] The term "asthma" above refers to a respiratory disease characterized by chronic airway inflammation, airway and bronchial hypersensitivity, and reversible airway obstruction, accompanied by symptoms such as wheezing, shortness of breath, chest tightness, and coughing, and can be classified into atopic asthma, allergic asthma, eosinophilic asthma, etc.

[0026] The term "subject" above refers to an individual that provides a sample for measuring the expression level of the mRNA of the EDN gene or the EDN protein, and ultimately serves as the subject of analysis for therapeutic responsiveness to an antibody therapeutic agent for asthma. The subject includes, without limitation, humans, mice, rats, guinea pigs, dogs, cats, horses, cattle, pigs, monkeys, chimpanzees, baboons, or rhesus monkeys, and specifically, is a human. Since the composition of the present invention provides information for predicting not only the therapeutic responsiveness but also the prophylactic responsiveness of the antibody therapeutic agent for asthma, the subject of the present invention may be an asthma patient or a healthy subject that has not yet been confirmed to have asthma.

[0027] The term "mRNA (messenger RNA)" refers to RNA that acts as a blueprint for polypeptide synthesis (protein translation) by transmitting the genetic information of a specific gene's base sequence to the ribosome during the protein synthesis process. Single-stranded mRNA is synthesized through the process of transcription using the gene as a template.

[0028] The term "protein" above is used interchangeably with "polypeptide" and refers to a polymer of amino acid residues, for example, as generally found in proteins in their natural state.

[0029] In the present invention, the agent for measuring the expression level of the mRNA of the EDN gene may be at least one selected from the group consisting of an antisense oligonucleotide that binds complementarily to the mRNA of the EDN gene, a primer, and a probe.

[0030] In the present invention, measuring the expression level of mRNA is a process of confirming the presence and degree of expression of mRNA of the EDN gene in a biological sample to predict the therapeutic responsiveness of a subject to an asthma antibody treatment, and can be determined by measuring the amount of mRNA of the EDN gene. Analysis methods for this purpose include RT-PCR, competitive RT-PCR, real-time RT-PCR, RNase protection assay (RPA), northern blotting, and DNA microarray chips, but are not limited to these.

[0031] The term "antisense nucleotide" refers to a nucleic acid-based molecule that has a sequence complementary to a target gene or polynucleotide and can form a dimer with the target gene or polynucleotide, and can be used to detect the target gene. The antisense polynucleotide may have an appropriate length to increase detection specificity.

[0032] The aforementioned "primer" is a short single-strand oligonucleotide that acts as a starting point for DNA synthesis. Under suitable buffer and temperature conditions, the primer specifically binds to a polynucleotide serving as a template, and DNA synthesis occurs when DNA polymerase adds a nucleoside triphosphate containing bases complementary to the template DNA to the primer. Primers generally consist of 15 to 30 base sequences, and the melting temperature (Tm) at which they bind to the template strand varies depending on their base composition and length. The primer sequence does not need to be completely complementary to a portion of the template's base sequence; it is sufficient to possess adequate complementarity within the range that allows the primer to hybridize with the template and perform its inherent function. Therefore, in the present invention, the primers for measuring the expression level of the EDN mRNA do not need to have a sequence that is perfectly complementary to each gene sequence; it is sufficient if they have a length and complementarity suitable for the purpose of measuring the amount of mRNA by amplifying a specific segment of mRNA or cDNA through DNA synthesis. The primers for the amplification reaction consist of a set (pair) that binds complementarily to the template (or sense) and the opposite side (antisense) at both ends of the specific segment of the mRNA to be amplified, respectively. A person skilled in the art can easily design the primers by referring to the mRNA or cDNA base sequence of the EDN. In the present invention, the therapeutic responsiveness of a subject to an asthma antibody therapeutic agent can be predicted by determining whether the PCR product is amplified after performing PCR amplification using forward and reverse primers for the EDN gene.

[0033] The term "probe" above refers to a fragment of a polynucleotide, such as RNA or DNA, ranging in length from a few to hundreds of base pairs, capable of specifically binding to the mRNA, cDNA (complementary DNA), or DNA of a specific gene. It is labeled to allow verification of the presence or expression level of the target mRNA or cDNA to be bound. The selection of the probe and hybridization conditions can be appropriately selected according to techniques known in the art. The probe may be used in diagnostic methods for detecting alleles (or alleles). Such diagnostic methods include detection methods based on nucleic acid hybridization, such as Southern blot, and may be provided in a form pre-bound to the substrate of a DNA chip in methods using DNA chips. In the present invention, after performing hybridization using a probe complementary to the EDN gene, the therapeutic responsiveness of a subject to an asthma antibody treatment can be predicted based on whether hybridization has occurred. The selection of a suitable probe and hybridization conditions may be modified based on those known in the art.

[0034] In the present invention, the antisense oligonucleotide, primer, or probe may be chemically synthesized using a phosphoramidite solid support synthesis method or other widely known methods. Additionally, the antisense oligonucleotide, primer, or probe may be modified in various ways according to methods known in the art, to the extent that hybridization with the target polynucleotide to be detected is not interfered with. Examples of such modifications include methylation, capping, substitution with one or more homologues of the natural nucleotide, and modification between nucleotides, such as uncharged linkages (e.g., methyl phosphonate, phosphotriester, phosphoroamidate, carbamate, etc.) or charged linkages (e.g., phosphorothioate, phosphorodithioate, etc.), and the binding of a labeling material using fluorescence or enzymes.

[0035] In the present invention, the antisense oligonucleotide, primer, or probe is not limited to a specific sequence as long as it is capable of measuring the expression of an EDN gene or mRNA, and can preferably be designed using known knowledge and techniques in the art.

[0036] In the present invention, the agent for measuring the expression level of the EDN protein may be at least one selected from the group consisting of antibodies, peptides, aptamers, and compounds that specifically bind to the EDN protein.

[0037] In the present invention, measuring the expression level of a protein is a process of confirming the presence and expression level of a protein encoded by the EDN gene in a biological sample to predict the therapeutic responsiveness of a subject to an asthma antibody treatment, and this can be determined by measuring the amount of the EDN protein. Analytical methods for this purpose include, but are not limited to, western blotting, ELISA (enzyme-linked immunosorbent assay), radioimmunoassay, radioimmunodiffusion, Ouchterlony immunodiffusion, Rocket immunoelectrophoresis, immunohistochemical staining, immunoprecipitation assay, complete fixation assay, FACS, and protein chip.

[0038] The term "antibody" above refers to a specific protein molecule induced for an antigenic site. In the present invention, the antibody refers to an antibody that specifically binds to an EDN protein, and the antibody may be prepared according to conventional methods in the art. In the present invention, the form of the antibody is not particularly limited and may include polyclonal antibodies, monoclonal antibodies, or parts thereof as long as they have antigen-binding ability, all immunoglobulin antibodies, and special antibodies such as humanized antibodies. For example, the antibody may include not only a complete form having two full-length light chains and two full-length heavy chains, but also functional fragments of the antibody molecule. A functional fragment of the antibody molecule refers to a fragment possessing at least an antigen-binding function, and may be, for example, Fab, F(ab'), F(ab')2, Fv, etc., but is not limited thereto.

[0039] Since the above-mentioned EDN protein is a known protein, the antibody used in the present invention can be prepared using the protein as an antigen by conventional methods widely known in the field of immunology. The EDN protein used as an antigen for the antibody according to the present invention can be extracted from nature or synthesized, and can be prepared by a recombinant method based on a DNA sequence. When using genetic recombination technology, the EDN protein can be obtained by inserting a nucleic acid encoding the EDN protein into a suitable expression vector, culturing host cells so that the EDN protein is expressed in a transformant transformed with the recombinant expression vector, and then recovering the EDN protein from the transformant.

[0040] In a biological sample, the EDN protein and the antibody conjugate specific thereto form an antigen-antibody complex, i.e., an antigen-antibody complex, and the amount of antigen-antibody complex formed can be quantitatively measured through the magnitude of the signal of a detection label. Such detection labels may be selected from a group consisting of enzymes, fluorescent dyes, ligands, luminescent substances, microparticles, redox molecules, and radioisotopes, but are not limited thereto. Analytical methods for measuring protein levels include, but are not limited to, Western blot, ELISA, radioimmunoassay, radioimmunodiffusion, Ouchteroni immunodiffusion, rocket immunoelectrophoresis, immunohistochemistry, immunoprecipitation analysis, complement fixation analysis, FACS, protein chips, etc. In the present invention, after conjugating an antibody against the EDN protein, the therapeutic responsiveness of the subject to an asthma antibody treatment can be predicted by quantifying the signal intensity using the ELISA method, etc.

[0041] The term "peptide" mentioned above has the advantage of high binding affinity to target substances and does not undergo denaturation even during heat or chemical treatment. Additionally, due to its small molecular size, it can be attached to other proteins to be used as a fusion protein.

[0042] The term "aptamer" above refers to a type of polynucleotide composed of a special kind of single-stranded nucleic acid (DNA, RNA, or modified nucleic acid) that has a stable tertiary structure in itself and is capable of binding to target molecules with high affinity and specificity. As described above, aptamers can specifically bind to antigenic substances just like antibodies, but because they are composed of polynucleotides that are more stable than proteins, have a simple structure, and are easy to synthesize, they can be used as a substitute for antibodies.

[0043] In the present invention, the asthma antibody therapeutic agent refers to a biological agent having an improving or therapeutic activity against asthma, and may include mepolizumab, reslizumab, dupilumab, omalizumab, anlukinumab, tralokinumab, quillizumab, benralizumab, imatinib, lebrikizumab, etc.

[0044] In particular, the above asthma antibody therapeutic agent may be at least one selected from the group consisting of mepolizumab, reslizumab, and dupilumab, which have improving or therapeutic activity for eosinophilic asthma or type 2 inflammatory asthma caused by elevated levels of eosinophils. The above mepolizumab and reslizumab are anti-IL-5 monoclonal antibodies targeting IL-5 or its receptor (IL-5R), and may have therapeutic activity for eosinophilic asthma by inhibiting the activity of IL-5 or its receptor. Meanwhile, the above dupilumab is an anti-IL-4 monoclonal antibody targeting IL-4 or its receptor (IL-4R), and may have therapeutic activity for eosinophilic asthma by inhibiting the activity of IL-4 or its receptor.

[0045] According to the present invention, if the expression level of the mRNA of the EDN gene or the EDN protein measured using the composition of the present invention is increased compared to the control group, the therapeutic responsiveness to the asthma antibody treatment can be evaluated as good.

[0046] For example, if the expression level of the mRNA of the EDN gene or the EDN protein is increased by 1% or more, 2% or more, 3% or more, 4% or more, 5% or more, 6% or more, 7% or more, 8% or more, 9% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, or 50% or more compared to the control group, the therapeutic response to the asthma antibody treatment can be evaluated as good.

[0047] The above control group may refer to normal subjects, a group of asthma patients not administered asthma antibody treatment, or the median (average value) of a group of asthma patients after administration of asthma antibody treatment.

[0048] The above term "therapeutic responsiveness" may mean that after administration of an asthma antibody therapeutic agent, asthma does not develop in the subject, the progression of asthma is slowed, asthma is improved, or therapeutic activity is enhanced, resulting in an increased survival rate of the subject.

[0049] The term "prediction" above means to estimate or guess in advance regarding medical outcomes, and for the purposes of the present invention, it means to estimate in advance the therapeutic responsiveness (disease progression, improvement, drug resistance) of a subject treated with an asthma antibody therapeutic agent.

[0050] The composition of the present invention allows for the selection of a treatment method suitable for each individual subject by predicting a patient group with high therapeutic responsiveness in advance before administering an asthma antibody therapeutic agent to the subject.

[0051] In the present invention, the expression level of the mRNA of the EDN gene or the EDN protein may be measured in a sample isolated from the subject.

[0052] The above sample may be used without limitation from a subject for whom therapeutic responsiveness to an asthma antibody treatment is to be predicted, and may be, for example, cells or tissues obtained by biopsy, blood, whole blood, serum, plasma, saliva, cerebrospinal fluid, various secretions, urine, feces, etc. Specifically, the above sample may be at least one selected from the group consisting of tissue, cells, cell extracts, plasma, serum, blood, saliva, mucus, semen, nasal aspirate, bronchial aspirate, organ secretions, meningeal fluid, cerebrospinal fluid, and urine. More specifically, the above sample may be plasma, serum, blood, or urine. The above sample may be pretreated before use for detection or diagnosis. For example, this may include homogenization, filtration, distillation, extraction, concentration, inactivation of interfering components, addition of reagents, etc.

[0053] According to the present invention, if the expression level of the mRNA of the EDN gene or the EDN protein measured in the sample using the composition of the present invention is increased compared to the control group, the therapeutic responsiveness to the asthma antibody therapeutic agent can be evaluated as good.

[0054] In the present invention, the composition may further include a preparation for measuring the number of blood eosinophils.

[0055] The above-mentioned eosinophil is a type of pro-inflammatory white blood cell (WBC) characterized by a multilobed nucleus and cytoplasmic granules that are easily stained by eosin, and is known to play an important role in airway inflammation.

[0056] The term "blood eosinophil count" refers to the number of eosinophils in the blood, which can be measured, for example, by counting the number of eosinophils per 100 cells and multiplying the resulting percentage by the white blood cell count (i.e., total white blood cell count), and is typically calculated per μL of blood or per mm of blood. 3 It can be expressed as the number of sugar cells (i.e., eosinophils).

[0057] The aforementioned eosinophils are known to play an important role in airway inflammation, particularly in the development of eosinophilic asthma. Accordingly, blood eosinophil count parameters, which count the number of these eosinophils, have previously been used to predict treatment responsiveness for asthma, specifically eosinophilic asthma.

[0058] If the composition of the present invention further comprises a preparation for measuring the number of blood eosinophils, and the AUC value of at least one of the parameters a) to h) below is higher than the AUC value of the reference number of blood eosinophils, the therapeutic responsiveness to the antibody therapeutic agent can be evaluated as good:

[0059] a) Serum EDN decreased ratio from baseline to 1 month

[0060] b) Serum EDN level / blood eosinophil count at baseline

[0061] c) Serum EDN level / blood eosinophil count at 1 month

[0062] d) ΔSerum EDN / ΔBlood eosinophil count at 1 month

[0063] e) Adjusted Urine EDN decreased ratio compared to baseline at 1 month

[0064] f) Adjusted Urine EDN / Blood eosinophil at baseline

[0065] g) Adjusted Urine EDN / Blood eosinophil count at 1 month

[0066] h) Adjusted ΔUrine EDN / ΔBlood eosinophil at 1 month.

[0067] The parameters of a) to h) above can each be calculated in the following manner:

[0068] a) Percentage decrease in serum EDN levels from baseline to 1 month: (Serum EDN level at 1 month - Serum EDN level at baseline) / Serum EDN level at baseline ((EDN 1month -EDN baseline ) / EDN baseline )

[0069] b) Serum EDN level / blood eosinophil count at reference: Serum EDN level / blood eosinophil count at reference (EDN baseline / BE baseline )

[0070] c) Serum EDN levels / blood eosinophil count at 1 month: Serum EDN levels / blood eosinophil count at 1 month (EDN 1month / BE 1month )

[0071] d) Δserum EDN level at 1 month / Δblood eosinophil count: |serum EDN level at baseline - serum EDN level at 1 month| / |blood eosinophil count at baseline - blood eosinophil count at 1 month| (|EDN baseline -EDN 1month | / |BE baseline -BE 1month |)

[0072] e) Percentage reduction in adjusted urinary EDN levels from baseline to 1 month: (Urinary EDN levels adjusted at 1 month - Urinary EDN levels adjusted at baseline) / Urinary EDN levels adjusted at baseline ((uEDN 1month -uEDN baseline ) / uEDN baseline )

[0073] f) Reference-adjusted urinary EDN levels / Blood eosinophil count: Reference-adjusted urinary EDN levels / Reference blood eosinophil count (uEDN baseline / BE baseline )

[0074] g) Adjusted urinary EDN level at 1 month / Blood eosinophil count: Adjusted urinary EDN level at 1 month / Blood eosinophil count at 1 month (uEDN 1month / BE 1month )

[0075] h) Adjusted ΔUrine EDN at 1 month / ΔBlood Eosinophil Count: |Adjusted Urine EDN at reference - Adjusted Urine EDN at 1 month| / |Blood Eosinophil Count at reference - Blood Eosinophil Count at 1 month| (|uEDN baseline -uEDN 1month | / |BE baseline -BE 1month |)

[0076] (Here, the adjusted urinary EDN level is determined by adjusting for serum creatinine levels.)

[0077] For example, if the AUC value of at least one of the parameters in a) to h) above is 1% or more, 2% or more, 3% or more, 4% or more, 5% or more, 6% or more, 7% or more, 8% or more, 9% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, or 50% or more higher than the AUC value of the reference blood eosinophil count, the therapeutic response to the asthma antibody treatment can be evaluated as good.

[0078] The above AUC (Area under the plasma level-time curve) value can be calculated by measuring serum EDN levels, urine EDN levels, and blood eosinophil counts from samples separated from the subject, and configuring them with various parameters.

[0079] In one embodiment of the present invention, serum EDN levels, urine EDN levels, and blood eosinophil counts were measured in asthma patients treated with biological agents such as mepolizumab, reslizumab, or dupilumab. Subsequently, eight parameters were constructed using these parameters to measure AUC values, which were then compared with the AUC values ​​of blood eosinophil counts, which are conventionally used to predict therapeutic responsiveness to asthma antibody treatments. As a result, it was confirmed that the AUC values ​​for responders or initial responders to the antibody treatments were higher for several parameters compared to the blood eosinophil count parameter, indicating superior efficiency in predicting therapeutic responsiveness to asthma antibody treatments. Thus, the present invention can be usefully utilized for predicting the therapeutic responsiveness of subjects to asthma antibody treatments.

[0080]

[0081] 2. Kit for predicting a subject's therapeutic responsiveness to asthma antibody treatments

[0082] Another aspect of the present invention provides a kit capable of predicting the therapeutic responsiveness of a subject to an asthma antibody treatment.

[0083] The overlapping description is identical to that described above in '1. Composition for predicting the therapeutic responsiveness of a subject to an asthma antibody therapeutic agent,' and therefore is omitted.

[0084] In the present invention, the kit comprises as an active ingredient a preparation for measuring the expression level of the mRNA of the EDN (Eosinophil-derived neurotoxin) gene or the EDN protein.

[0085] The agent for measuring the expression level of the mRNA of the EDN gene may be at least one selected from the group consisting of an antisense oligonucleotide, a primer, and a probe that binds complementarily to the EDN mRNA, and the agent for measuring the expression level of the EDN protein may be at least one selected from the group consisting of an antibody, a peptide, an aptamer, and a compound that specifically binds to the EDN protein.

[0086] The above asthma antibody treatment may be at least one selected from the group consisting of mepolizumab, reslizumab, and dupilumab.

[0087] In the present invention, the expression level of the mRNA of the EDN gene or the EDN protein may be measured in a sample isolated from the subject.

[0088] According to the present invention, if the expression level of the mRNA of the EDN gene or the EDN protein measured in the sample using the kit of the present invention is increased compared to the control group, the therapeutic responsiveness to the asthma antibody treatment can be evaluated as good.

[0089] In the present invention, the kit may further include a preparation for measuring the number of blood eosinophils, and in this case, if the AUC value of at least one of the parameters a) to h) described above in '1. Composition for predicting the therapeutic responsiveness of a subject to an asthma antibody therapeutic agent' is higher than the AUC value of the reference blood eosinophil number, the therapeutic responsiveness to the asthma antibody therapeutic agent can be evaluated as good.

[0090] In the present invention, the kit may further include an instruction manual. The instruction manual may include information regarding data interpretation and judgment, and may state criteria such that the therapeutic response to the asthma antibody treatment can be evaluated as good if, for example, the expression level of the mRNA of the EDN gene or the EDN protein is increased compared to the control group, or if the AUC value of at least one of the parameters of a) to h) described above is higher than the AUC value of the reference blood eosinophil count.

[0091] In the present invention, the kit may be an RT-PCR kit, a DNA chip kit, an ELISA kit, a protein chip kit, a rapid kit, or an MRM (Multiple reaction monitoring) kit.

[0092] The kit of the present invention may further include one or more other component compositions, solutions, or devices suitable for the analysis method. For example, the kit of the present invention may further include essential elements necessary to perform a reverse transcription polymerase chain reaction. The reverse transcription polymerase chain reaction kit includes a pair of primers specific to a gene encoding a marker protein. The primers are nucleotides having a sequence specific to the nucleic acid sequence of said gene and may have a length of about 7 bp to 50 bp, more preferably about 10 bp to 30 bp. It may also include primers specific to the nucleic acid sequence of a control gene. Furthermore, the reverse transcription polymerase chain reaction kit may include a test tube or other suitable container, a reaction buffer (with varying pH and magnesium concentration), deoxyribonucleotides (dNTPs), enzymes such as Taq-polymerase and reverse transcription enzymes, DNase, RNase inhibitor DEPC-water, sterile water, etc.

[0093] In addition, the kit of the present invention may include essential elements necessary for performing DNA chip operations. The DNA chip kit may include a substrate to which cDNA or oligonucleotides corresponding to a gene or a fragment thereof are attached, and reagents, preparations, enzymes, etc., for producing fluorescently labeled probes. Additionally, the substrate may include cDNA or oligonucleotides corresponding to a control gene or a fragment thereof.

[0094] In addition, the kit of the present invention may include essential elements necessary for performing an ELISA. The ELISA kit includes an antibody specific to the EDN protein. The antibody is a monoclonal antibody, a polyclonal antibody, or a recombinant antibody that has high specificity and affinity for the marker protein and has little cross-reactivity with other proteins. Additionally, the ELISA kit may include an antibody specific to a control protein. Furthermore, the ELISA kit may include reagents capable of detecting the bound antibody, such as a labeled secondary antibody, chromophores, an enzyme (e.g., conjugated with the antibody) and its substrate, or other substances capable of binding to the antibody.

[0095] In the kit of the present invention, a nitrocellulose membrane, a PVDF membrane, a well plate synthesized from polyvinyl resin or polystyrene resin, a glass slide glass, etc. may be used as a fixative for the antigen-antibody binding reaction, but is not limited thereto.

[0096] In addition, the label of the secondary antibody in the kit of the present invention is preferably a conventional chromogenic agent that produces a color reaction, and may be labeled with fluorescent substances (fluorescein) and dyes such as HRP (horseradish peroxidase), alkaline phosphatase, colloid gold, and RITC (rhodamine-B-isothiocyanate), but is not limited thereto.

[0097] In addition, the chromogenic substrate for inducing color development in the kit of the present invention is preferably used according to the label that performs the color reaction, and TMB (3,3',5,5'-tetramethylbezidine), ABTS [2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)], OPD (o-phenylenediamine), etc. may be used. At this time, it is more preferable that the chromogenic substrate be provided in a state dissolved in a buffer solution (0.1 M NaAc, pH 5.5). A chromogenic substrate such as TMB is degraded by HRP used as a label for the secondary antibody conjugate to produce a chromogenic precipitate, and the presence or absence of the marker proteins is detected by visually confirming the degree of precipitation of this chromogenic precipitate.

[0098] In the kit of the present invention, the washing solution may include phosphate buffer solution, NaCl, and Tween 20, and the washing solution may be added to the immobilizer in an appropriate amount and performed 3 to 6 times after reacting a secondary antibody with the antigen-antibody conjugate following the antigen-antibody binding reaction.

[0099]

[0100] 3. Method for Predicting the Treatment Response of Subjects to Asthma Antibody Therapeutics

[0101] Another aspect of the present invention provides a method for predicting the therapeutic responsiveness of a subject to an asthma antibody therapeutic agent.

[0102] The overlapping description is identical to that described above in '1. Composition for predicting the therapeutic responsiveness of a subject to an asthma antibody therapeutic agent,' and therefore is omitted.

[0103] In the present invention, the method comprises the step of measuring the expression level of the mRNA of the EDN (Eosinophil-derived neurotoxin) gene or the EDN protein in a sample separated from a subject.

[0104] The measurement of the expression level of the EDN mRNA can be performed using at least one selected from the group consisting of antisense oligonucleotides, primers, and probes that bind complementarily to the EDN mRNA, and the measurement of the expression level of the EDN protein can be performed using at least one selected from the group consisting of antibodies, peptides, aptamers, and compounds that specifically bind to the EDN protein.

[0105] At this time, the expression level of the mRNA of the EDN gene or the EDN protein is in a group consisting of RT-PCR, competitive RT-PCR, real-time RT-PCR, quantitative RT-PCR (reverse transcription polymerase chain reaction), RNase protection assay (RPA), Northern blotting, DNA chip analysis, western blotting, enzyme-linked immunoabsorbent assay (ELISA), ligand binding assay, radioimmunoassay, radioimmunodiffusion, Ouchterlony immunodiffusion, Rocket immunoblot analysis, immunoelectrophoresis, immunohistochemical staining, immunoprecipitation assay, complete fixation assay, FACS, and protein chip analysis It may be measured by at least one selected method.

[0106] In the method of the present invention, the asthma antibody therapeutic agent may be at least one selected from the group consisting of mepolizumab, reslizumab, and dupilumab.

[0107] In the method of the present invention, the sample may be plasma, serum, blood, or urine.

[0108] In this case, if the expression level of the mRNA of the EDN gene or the EDN protein measured in the above sample is higher than that of the control group, it can be evaluated that the therapeutic response to the asthma antibody treatment is good.

[0109] For example, if the expression level of the mRNA of the EDN gene or the EDN protein is increased by 1% or more, 2% or more, 3% or more, 4% or more, 5% or more, 6% or more, 7% or more, 8% or more, 9% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, or 50% or more compared to the control group, the therapeutic response to the asthma antibody treatment can be evaluated as good.

[0110] Meanwhile, the method of the present invention may further include the step of measuring the number of blood eosinophils.

[0111] If the method of the present invention further includes the step of measuring the number of blood eosinophils, and the AUC value of at least one of the parameters a) to h) below is higher than the AUC value of the reference number of blood eosinophils, the therapeutic responsiveness to the asthma antibody therapeutic agent can be evaluated as good:

[0112] a) Serum EDN decreased ratio from baseline to 1 month

[0113] b) Serum EDN level / blood eosinophil count at baseline

[0114] c) Serum EDN level / blood eosinophil count at 1 month

[0115] d) ΔSerum EDN / ΔBlood eosinophil count at 1 month

[0116] e) Adjusted Urine EDN decreased ratio compared to baseline at 1 month

[0117] f) Adjusted Urine EDN / Blood eosinophil at baseline

[0118] g) Adjusted Urine EDN / Blood eosinophil count at 1 month

[0119] h) Adjusted ΔUrine EDN / ΔBlood eosinophil at 1 month.

[0120] For example, if the AUC value of at least one of the parameters in a) to h) above is 1% or more, 2% or more, 3% or more, 4% or more, 5% or more, 6% or more, 7% or more, 8% or more, 9% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, or 50% or more higher than the AUC value of the reference blood eosinophil count, the therapeutic response to the asthma antibody treatment can be evaluated as good.

[0121] In the present invention, if the expression level of the mRNA of the EDN gene or the EDN protein measured in a sample isolated from the subject is higher than that of the control group, and at the same time the AUC value of at least one of the parameters a) to h) is higher than that of the reference blood eosinophil count, the therapeutic responsiveness to the asthma antibody therapeutic agent can be evaluated as being better.

[0122]

[0123] The present invention will be explained in detail below through examples.

[0124] However, the following examples are intended to specifically illustrate the present invention, and the content of the present invention is not limited by the following examples.

[0125]

[0126] [Example]

[0127] Example 1. Experimental Method

[0128] 1-1. Target

[0129] A total of 56 patients diagnosed with severe asthma were analyzed. These patients received mepolizumab (13 patients, 23.2%), reslizumab (23 patients, 41.1%), or dupilumab (20 patients, 35.7%) formulations from April 2020 to May 2021. Among the patients, 29 (51.8%) were male, the average age was 53.48 ± 10.83 years, and the average duration of asthma was 11.68 ± 8.41 years. 35 patients (62.5%) experienced acute exacerbations in the past 12 months, with an average of 4.41 ± 6.36 exacerbations, and 19 patients (33.9%) used mOCS. 42 (75%) and 30 (53.6%) patients exhibited allergic rhinitis and chronic rhinitis, respectively. To evaluate the therapeutic response of patients to the above biological agents, they were observed for 24 weeks.

[0130]

[0131] 1-2. Measurement of Serum and Urine EDN (Eosinophil-derived Neurotoxin) Levels

[0132] Serum EDN levels were measured in airway secretion samples collected via sterile nebulized saline inhalation. Serum and urinary EDN levels were measured using the K EDN ELISA kit (SKIMS-BIO Co., Seoul, Korea) within detection ranges of 6.0 and 400 ng / mL. Urinary EDN levels were adjusted by dividing the result of urine sample analysis by the serum creatinine level.

[0133]

[0134] 1-3. Classification of Treatment Responses

[0135] After 24 weeks of treatment, patients' responsiveness to the biological agent was evaluated based on maintenance oral corticosteroid dose and annualized exacerbation rate. Patients who demonstrated a therapeutic response were classified as responders or super responders, while those who did not respond were classified as nonresponders. Specifically, patients were classified as responders if their annualized exacerbation rate decreased by more than 50%, if they required maintenance oral corticosteroid (mOCS) therapy, or if their daily mOCS dose decreased by more than 50%. Patients who experienced no exacerbations and no longer required mOCS therapy were classified as super responders. Furthermore, if the annual exacerbation rate decreased by more than 50%, the FEV1 (forced expiratory volume in one second) volume increased by more than 100 mL after bronchodilator treatment, or the ACT (asthma control test) score increased by more than 3 points, the therapeutic response to each biological agent was evaluated as good.

[0136]

[0137] 1-4. Statistical Analysis

[0138] The correlation between reference serum and urine EDN levels and clinical characteristics of asthma was evaluated using the Wilcoxon rank-sum test and the Spearman correlation coefficient. The Kruskal-Wallis test was used to evaluate non-normally distributed variables, and log transformation was used to standardize asymmetric key exposures. T-tests were used to compare key exposure values ​​across various treatment response groups. Additionally, a multivariate logistic regression model was used to calculate the sensitivity, specificity, and AUC (area under the curve) of the ROC (receiver operating characteristic) curve for key exposure values, thereby identifying the most effective parameters for predicting treatment response. Statistical analysis was performed using SAS (SAS Institute v.9.4, Cary, NC), and a p-value ≤ 0.05 was considered statistically significant.

[0139]

[0140] Example 2. Experimental Results

[0141] 2-1. Confirmation of patient's serum and urine EDN levels

[0142] The patients' serum and urine EDN levels and blood eosinophil counts were measured at 1 month and 6 months of treatment, respectively (Table 1).

[0143] Patient Product Blood Eosinophils Transfusion Agency EDN Urine EDN Standard 1 Month 6 Month Standard 1 Month 6 Month Standard 1 Month 6 Month 1Res509.249.846.2131.3939.3525.1260.193.438.22Res445.930.111.7146.8632.7426.5120.465.8443Res18420.830.592.3242.5849.84222.8168.9417.94Res204.727.34942.7827.2238.214.523.3105Res928.8166.419.8230.8258.7529.785585.913.46R es567.610.419.2158.5228.7514.353349.534.57Res26.837.23343.665 3.4348.4822.140798Res117691.284287.7750.0848.390.898.585.29Re s62399.229.2101.6885.4821.9235.8160.233.610Res12164628.8322.7 60.5633.1699.910.351.111Res917.690.133203.668.3136.9383.9382.1 51.312Res341.717.176.886.1732.0948.9118.8246.550.813Res798.65 2.212203.2118.3863.83123.849.27.814Res1150.214458.880.7442.64 17.73120.9136.367.515Res9037831.2152.2735.1914.16125.8141.554 .816Res1532.2127.898.4257.2747.0233.72472.868.713.317Res149414 7.232.8347.3345.5925.421895.7275.515.118Res39641.440.888.6944 .1730.46154.977.429.719Res956.873.617.2165.0139.3931.4564.162 .92120Res108106.7214.444.6860.8137.0154.648.936.921Res345.619 .517.1126.1125.1221.79110.2126.610.722Res671.617468.9128.6446.6544.14123.7141.613.623Res162858.280.4308.1940.9544.5984.63221.824Mep2856102.630.4244.5659.7525.4535.9176.214.425Mep6827050.4140.8551.7435.3488.610.6926Mep27060446.6130.6857.8962.3849.249.652.927Mep456107.2100.5212.6655.2558.4671.3128.631.728Mep8198851228.5752.8856.66227.519410629Mep1394.4278.8438.9296.41296.41149.763.338.519.930Mep7109986.9218.361.671.469.770.260.931Mep525.1..76.2334.4344.78157.284.549.632Mep676.2..62.0321.7634.2295.430.418.833Mep804.6127.64157.4219.813.83231.690.684.234Mep393.68458.455.9118.35.58.169.4.35Mep579.611078124.8636.6130.3230.436.222.436Mep207.918786.465.8135.45.22.979.9.37Dup382.2323108977.8593.85167.6970.579.2262.938Dup481.8399.6284.2180.45166.19128.417.770.4133.739Dup3121793.525273.73227.41143.0627.927.3435.840Dup282.5597.8552144.95252.63155.2459.221.514541Dup202.422321781.9111.17315.72241.2412297.7403.842Dup249.4477410.4100.81139.05191.9315.9149.341.443Dup74.41079.7552.951.17220.11180.7244.2770.6168.544Dup74.424.419450.5648.2570.8936.765.619.845Dup7922249.1979.2289.36337.08267.228.5117.419346Dup9.4.754.6111.451 10.44174.4242.4210.882.747Dup295.2652.8598.493.99266.9259.5675.363.444 6.148Dup378.4317.4277.261.2362.8745.8643.449.819.449Dup4031659625.1101 .46264.47170.5363.5225.1211.650Dup163.39686.951.754.6647.1973.233.431.8 51Dup48.31638177.641.5188.443.1288.879.812.152Dup82.5210.610030.9260.5 143.8938.688.162.853Dup497885.6270.176.09149.1685.4358.962.361.254Dup4 41.63059.2828.296.02250.4250.9986.5135.271.255Dup570817.81829.759.3154 .54115.0398.354.9201.956Dup4509.1361.8247.2875.24168.92266.4142.8609.1.

[0144]

[0145] In the mepolizumab (Mep) and reslizumab (Res) treatment groups, blood eosinophil counts, serum EDN levels, and urine EDN levels all showed a decreasing trend as the treatment period progressed (Fig. 1). In contrast, in the dupilumab (Dup) treatment group, blood eosinophil counts and serum EDN levels showed a tendency to increase and then decrease depending on the patient, while urine EDN levels showed a tendency to increase as the treatment period progressed (Fig. 1).

[0146]

[0147] 2-2. Analysis of the Correlation Between Serum and Urinary EDN Levels and Clinical Characteristics of Asthma

[0148] The correlation between patients' serum and urine EDN levels and the clinical characteristics of asthma was analyzed (Table 2).

[0149] Serum EDN Urine EDN Correlation coefficient p-value Correlation coefficient p-value Age (y) - 0.04 40.749 - 0.23 60.080 BMI - 0.472 <.00 1 - 0.35 30.008 Age of symptom onset (years) - 0.10 30.462 - 0.16 60.235 Duration of asthma (years) - 0.15 30.27 40.03 90.779 Number of exacerbations (previous 6 months) - 0.15 00.26 90.025 0.855 Reference ACT score - 0.10 70.437 - 0.30 40.024 Pre-BD FEV1 pred (%) Pre-BD FEV1 / FVC ratio - 0.16 90.218 - 0.06 70.627 Post-BD FEV1 pred (%) 0.02 0.883 - 0.02 7 0.843 Post-BD FEV1 / FVC Ratio - 0.10 7 0.454 - 0.00 1 0.997 Blood Eosinophils ( / μL) - 0.10 6 0.380 - 0.01 9 0.892 Sputum Eosinophils (%) FeNO (ppb) 0.72 9 <.00 1 0.45 3 0.001 Total IgE (kU / L) 0.32 1 0.04 3 0.39 2 0.012

[0150]

[0151] Serum and urine EDN levels were found to have a negative correlation with BMI (serum EDN level: -0.472, p=<.0001; urine EDN level: -0.353, p=0.008). Additionally, urine EDN levels showed a negative correlation with the reference ACT score (-0.304, p=0.024). Both serum and urine EDN levels had a positive correlation with blood eosinophil count ( / μL) and sputum eosinophils (%); a moderate correlation was observed between serum EDN levels and blood eosinophil count ( / μL) (0.729, p=<.0001), while a weak correlation was found between urine EDN levels (0.453, p=0.001) (Figure 1). Sputum eosinophils (%) showed a weak correlation with both serum and urine EDN levels (serum EDN level: 0.321, p=0.043; urine EDN level: 0.392, p=0.012). For fractional exhaled nitric oxide (FeNO) (ppb) and total IgE (kU / L), no significant correlation was observed with serum and urine EDN levels.

[0152]

[0153] 2-3. Prediction of Treatment Response to Asthma Antibody Therapeutics Using Serum and Urinary EDN Levels

[0154] Logistic multivariate regression analysis and ROC curve analysis were performed to determine whether serum and urine EDN levels could predict treatment responsiveness when treating asthma patients with antibody therapy. In this case, various parameters were set using the blood eosinophil count, which is commonly used as a marker for predicting treatment responsiveness, along with the two values ​​(Table 3).

[0155] Parameter Definition Criteria Serum EDN Level (Baseline Serum EDN) EDN baseline Serum EDN decreased ratio from baseline to 1 month (EDN 1month-EDN baseline ) / EDN baseline Serum EDN / Blood eosinophil count at baseline baseline / BE baseline Serum EDN level / Blood eosinophil count at 1 month 1month / BE 1month Serum EDN level / Blood eosinophil count at 1 month | EDN baseline -EDN 1month | / |BE baseline -BE 1month |Baseline urine EDN level (Baseline urine EDN) uEDN baseline Adjusted Urine EDN decreased ratio compared to baseline at 1 month (uEDN) 1month -uEDN baseline ) / uEDN baseline Adjusted Urine EDN / Blood eosinophil at baseline baseline / BE baseline Adjusted Urine EDN / Blood eosinophil count at 1 month uEDN 1month / BE 1month Adjusted Urine EDN / Blood eosinophil at 1 month | uEDN baseline -uEDN 1month | / |BE baseline -BE 1month |

[0156]

[0157] Subsequently, treatment responders were classified into five groups based on the criteria listed in Table 4 below, and to evaluate the predictability for each group, the AUC of the parameter using serum EDN levels among the parameters listed in Table 3 above was compared with the AUC of the reference blood eosinophil count (Table 5).

[0158] Definition of Treatment Response Responder: Annual exacerbation rate decreases by 50% or more, or, for patients requiring long-term oral corticosteroid (mOCS) therapy, daily mOCS dose decreases by 50% or more Super responder: No exacerbations occur and mOCS therapy is no longer required Exacerbation: Annual exacerbation rate decreases by 50% or more ACT scores: Improvement of 3 points or more Pre-bronchodilator FEV1: Dose of 100 mL or more and an increase of 10% or more

[0159]

[0160] First, in the entire patient group, serum EDN levels / blood eosinophil counts and baseline FeNO parameters at 1 month showed higher AUC values ​​than the baseline blood eosinophil counts in responder prediction (0.875 and 0.872 versus 0.848, respectively). The rate of decline in serum EDN levels from baseline to 1 month showed higher AUC values ​​than the baseline blood eosinophil counts in initial responder prediction and FEV1 responder prediction before bronchodilator treatment at 6 months (0.805 versus 0.780, and 0.804 versus 0.752, respectively). Furthermore, the change in serum EDN level / change in blood eosinophil count parameter from baseline to 1 month (Δserum EDN level / Δblood eosinophil count at 1 month) showed a higher AUC than the reference blood eosinophil count AUC value in ACT score and FEV1 responder prediction before bronchodilator treatment (0.867 vs. 0.841, and 0.785 vs. 0.752, respectively).

[0161] In addition, in the patient groups treated with mepolizumab or reslizumab, the parameter of change in serum EDN levels / change in blood eosinophil count from baseline to 1 month showed a higher AUC than the baseline blood eosinophil count for predicting treatment responders in all five categories. With the exception of the baseline serum EDN levels / blood eosinophil count parameter, all parameters showed higher AUC values ​​than the baseline blood eosinophil count for predicting FEV1 responders before bronchodilator treatment. Furthermore, at 1 month, the serum EDN levels / blood eosinophil count parameter was found to have the highest AUC value for responder prediction (0.845), and the parameter of change in serum EDN levels / change in blood eosinophil count from baseline to 1 month was found to have the highest AUC value for predicting FEV1 responders before bronchodilator treatment (0.716).

[0162] In addition, in the dupilumab patient group, the parameter of change in serum EDN levels / change in blood eosinophil count from baseline to 1 month was found to have the highest AUC value for all predictions except responder prediction. Meanwhile, the parameter of the rate of decrease in serum EDN levels from baseline to 1 month showed the highest AUC value for responder prediction (0.954).

[0163] Next, the AUC of the parameter using urine EDN levels among the parameters listed in Table 3 above was compared with the AUC of the reference blood eosinophil count (Table 6).

[0164]

[0165] In the entire patient group, for baseline-adjusted urinary EDN levels / blood eosinophil count parameters, a higher AUC was observed compared to the AUC value of the baseline blood eosinophil count when predicting responders, super responders, and deteriorating responders.

[0166] In addition, the patient groups treated with mepolizumab or reslizumab showed a higher AUC than the reference blood eosinophil count AUC value when predicting FEV1 responders prior to bronchodilator treatment for all parameters.

[0167] In addition, in the dupilumab-treated patient group, all parameters showed an AUC higher than the reference blood eosinophil count AUC value when predicting responders and super responders.

[0168]

[0169] Although representative embodiments of the present invention have been described above by way of example, the scope of the present invention is not limited to such specific embodiments, and those skilled in the art will be able to make appropriate modifications within the scope described in the claims of this application.

Claims

A composition for predicting the therapeutic responsiveness of a subject to an asthma antibody therapeutic agent, comprising as an active ingredient a preparation for measuring the expression level of the mRNA or EDN (Eosinophil-derived neurotoxin) gene or EDN protein. In claim 1, A composition in which a preparation for measuring the expression level of the mRNA of the EDN gene is at least one selected from the group consisting of an antisense oligonucleotide that binds complementarily to the mRNA of the EDN gene, a primer, and a probe. In claim 1, A composition in which a preparation for measuring the expression level of the above EDN protein is at least one selected from the group consisting of antibodies, peptides, aptamers, and compounds that specifically bind to the EDN protein. In claim 1, A composition wherein the above asthma antibody therapeutic agent is at least one selected from the group consisting of mepolizumab, reslizumab, and dupilumab. In claim 1, A composition in which the above expression level is measured in a sample separated from the above subject. In claim 5, A composition wherein the sample is at least one selected from the group consisting of tissue, cell, cell extract, plasma, serum, blood, saliva, mucus, semen, nasal aspirate, bronchial aspirate, organ secretions, meningeal fluid, cerebrospinal fluid, and urine. In claim 6, A composition in which the above sample is plasma, serum, blood, or urine. In claim 7, A composition in which the above sample is urine. In claim 1, The above composition further comprises a preparation for measuring the number of blood eosinophils. A kit for predicting the therapeutic responsiveness of a subject to an asthma antibody therapeutic agent, comprising a composition of any one of claims 1 to 9. In claim 10, A kit that includes additional instruction manuals. A method for predicting the therapeutic responsiveness of a subject to an asthma antibody therapeutic agent, comprising the step of measuring the expression level of the mRNA of the EDN (Eosinophil-derived neurotoxin) gene or the EDN protein in a sample separated from the subject. In claim 12, A method in which the above asthma antibody therapeutic agent is at least one selected from the group consisting of mepolizumab, reslizumab, and dupilumab. In claim 12, A method in which the above sample is plasma, serum, blood, or urine. In claim 14, The above sample is urine, method. In claim 12, A method wherein the expression level is measured by at least one method selected from the group consisting of RT-PCR, competitive RT-PCR, real-time RT-PCR, reverse transcription polymerase chain reaction (RT-PCR), RNase protection assay (RPA), Northern blotting, DNA chip analysis, western blotting, enzyme-linked immunoabsorbent assay (ELISA), ligand binding assay, radioimmunoassay, radioimmunodiffusion, Ouchterlony immunodiffusion, Rocket immunoblot analysis, immunoelectrophoresis, immunohistochemical staining, immunoprecipitation assay, complete fixation assay, FACS, and protein chip analysis. In claim 12, A method for evaluating that therapeutic responsiveness to an asthma antibody treatment is good when the expression level of the mRNA of the EDN gene or the EDN protein measured in the above sample is higher than that of the control group. In claim 17, The above method further comprises the step of measuring the number of blood eosinophils. In claim 18, A method for evaluating that therapeutic responsiveness to an asthma antibody therapeutic agent is good when the AUC value of at least one of the parameters in a) to h) below is higher than the AUC value of the reference blood eosinophil count: a) Serum EDN decreased ratio from baseline to 1 month b) Serum EDN level / blood eosinophil count at baseline c) Serum EDN level / blood eosinophil count at 1 month d) ΔSerum EDN / ΔBlood eosinophil count at 1 month e) Adjusted Urine EDN decreased ratio compared to baseline at 1 month f) Adjusted Urine EDN / Blood eosinophil at baseline g) Adjusted Urine EDN / Blood eosinophil count at 1 month h) Adjusted ΔUrine EDN / ΔBlood eosinophil at 1 month.

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